NZ624021B2 - Heteroaryl pyridone and aza-pyridone compounds as inhibitors of btk activity - Google Patents

Heteroaryl pyridone and aza-pyridone compounds as inhibitors of btk activity Download PDF

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NZ624021B2
NZ624021B2 NZ624021A NZ62402112A NZ624021B2 NZ 624021 B2 NZ624021 B2 NZ 624021B2 NZ 624021 A NZ624021 A NZ 624021A NZ 62402112 A NZ62402112 A NZ 62402112A NZ 624021 B2 NZ624021 B2 NZ 624021B2
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pyridyl
hydroxymethyl
methyl
compound
oxo
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NZ624021A
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NZ624021A (en
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James John Crawford
Daniel Fred Ortwine
Binqing Wei
Wendy B Young
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F Hoffmann La Roche Ag
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Priority claimed from PCT/US2012/063194 external-priority patent/WO2013067274A1/en
Publication of NZ624021A publication Critical patent/NZ624021A/en
Publication of NZ624021B2 publication Critical patent/NZ624021B2/en

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    • A61K31/50—Pyridazines; Hydrogenated pyridazines
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    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
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    • A61K31/53—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with three nitrogens as the only ring hetero atoms, e.g. chlorazanil, melamine
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    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
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    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5383—1,4-Oxazines, e.g. morpholine ortho- or peri-condensed with heterocyclic ring systems
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    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
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Abstract

Disclosed are heteroaryl pyridone and aza-pyridone compounds of formula I useful for inhibiting Bruton's Tyrosine (Btk) kinase, wherein the substituents are as defined in the specification. Also disclosed are methods of using compounds of formula I for in vitro, in situ, and in vivo diagnosis, and treatment of disorders associated with Btk kinase including inflammation, immune disorders, and cancer. An example of a compound of formula (I) is: 2-(4-(hydroxymethyl)-5-(1-methyl-5-(5-(4-(oxetan-3-yl)piperazin-1-yl)pyridin-2-ylamino)-6-oxo-1,6-dihydropyridin-3-yl)pyridin-3-yl)-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one reatment of disorders associated with Btk kinase including inflammation, immune disorders, and cancer. An example of a compound of formula (I) is: 2-(4-(hydroxymethyl)-5-(1-methyl-5-(5-(4-(oxetan-3-yl)piperazin-1-yl)pyridin-2-ylamino)-6-oxo-1,6-dihydropyridin-3-yl)pyridin-3-yl)-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one

Description

HETEROARYL PYRIDONE AND AZA-PYRIDONE NDS AS TORS OF BTK ACTIVITY CROSS REFERENCE TO RELATED APPLICATIONS 5 This non-provisional application filed under 37 CFR §1.53(b), claims the t under 35 USC §119(e) of U.S. ional Application Serial No. 61/555,393 filed on 3 November 2011, which is orated by reference in entirety.
FIELD OF THE INVENTION The invention relates generally to compounds for treating disorders mediated by 10 Bruton’s Tyrosine Kinase (Btk) including inflammation, immunological, and cancer, and more specifically to compounds which inhibit Btk activity. Also described herein are methods of using the compounds for in vitro, in situ, and in vivo diagnosis or treatment of mammalian cells, or associated pathological conditions.
BACKGROUND OF THE INVENTION 15 Protein kinases, the largest family of human enzymes, ass well over 500 proteins. Bruton’s Tyrosine Kinase (Btk) is a member of the Tec family of tyrosine kinases, and is a regulator of early B-cell development as well as mature B-cell activation, signaling, and survival.
B-cell ing through the B-cell receptor (BCR) can lead to a wide range of 20 biological outputs, which in turn depend on the pmental stage of the B-cell. The magnitude and duration of BCR s must be precisely regulated. Aberrant BCR- mediated signaling can cause disregulated B-cell activation and/or the formation of pathogenic auto-antibodies leading to multiple autoimmune and/or inflammatory diseases.
Mutation of Btk in humans results in X-linked agammaglobulinaemia (XLA). This disease is 25 associated with the impaired maturation of s, diminished immunoglobulin production, compromised T-cell-independent immune responses and marked attenuation of the sustained calcium sign upon BCR stimulation. Evidence for the role of Btk in ic ers and/or autoimmune disease and/or inflammatory disease has been established in Btk-deficient mouse models. For example, in standard murine preclinical models of systemic lupus erythematosus 30 (SLE), Btk deficiency has been shown to result in a marked amelioration of disease progression. Moreover, Btk deficient mice can also be ant to developing collageninduced arthritis and can be less susceptible to Staphylococcus-induced arthritis. A large 1 body of evidence supports the role of B-cells and the humoral immune system in the enesis of autoimmune and/or inflammatory diseases. Protein-based therapeutics (such as Rituxan) developed to deplete B-cells, represent an approach to the treatment of a number of autoimmune and/or inflammatory diseases. Because of Btk's role in B-cell activation, 5 inhibitors of Btk can be useful as inhibitors of B-cell mediated pathogenic activity (such as autoantibody production). Btk is also sed in osteoclasts, mast cells and monocytes and has been shown to be important for the function of these cells. For example, Btk deficiency in mice is associated with ed IgE-mediated mast cell activation (marked diminution of TNF-alpha and other inflammatory cytokine release), and Btk deficiency in humans is 10 associated with greatly reduced pha production by activated monocytes.
Thus, tion of Btk activity can be useful for the treatment of allergic disorders and/or autoimmune and/or inflammatory diseases such as: SLE, rheumatoid tis, multiple vasculitides, idiopathic thrombocytopenic purpura (ITP), myasthenia gravis, allergic rhinitis, and asthma (Di Paolo et al (2011) Nature Chem. Biol. 7(1):41-50; Liu et al (2011) Jour. of 15 Pharm. and Exper. Ther. 338(1):154-163). In addition, Btk has been reported to play a role in apoptosis; thus, inhibition of Btk activity can be useful for cancer, as well as the treatment of B-cell lymphoma, leukemia, and other hematological malignancies. Moreover, given the role of Btk in osteoclast function, the tion of Btk activity can be useful for the treatment of bone disorders such as osteoporosis. Specific Btk inhibitors have been reported (Liu (2011) 20 Drug Metab. and Disposition 39(10):1840-1849; US 7884108, WO 2010/056875; US 7405295; US 7393848; WO 53121; US 7947835; US 2008/0139557; US 7838523; US 2008/0125417; US 2011/0118233; PCT/US2011/050034 “PYRIDINONES/PYRAZINONES, METHOD OF , AND METHOD OF USE THEREOF”, filed 31 Aug 2011; PCT/US2011/050013 “PYRIDAZINONES, METHOD OF MAKING, AND METHOD OF 25 USE THEREOF”, filed 31 Aug 2011; US Ser. No. 13/102720 “PYRIDONE AND AZAPYRIDONE COMPOUNDS AND S OF USE”, filed 6 May 2011).
SUMMARY OF THE ION The invention relates lly to Formula I, heteroaryl pyridone and ridone compounds with Bruton’s Tyrosine Kinase (Btk) modulating activity. 30 Formula I compounds have the structures: 2 I including stereoisomers, tautomers, or pharmaceutically acceptable salts thereof. The various substituents are d herein below.
One aspect of the invention is a pharmaceutical composition comprised of a a I 5 compound and a pharmaceutically acceptable carrier, glidant, diluent, or excipient. The ceutical composition may r comprise a second therapeutic agent.
Also described herein is a process for making a pharmaceutical composition which comprises combining a Formula I compound with a pharmaceutically acceptable carrier.
Also described herein is a method of treating a e or disorder which method 10 comprises administering a therapeutically effective amount of a Formula I compound to a patient with a disease or disorder selected from immune disorders, cancer, cardiovascular e, viral ion, inflammation, metabolism/endocrine function disorders and neurological disorders, and mediated by Bruton’s tyrosine kinase.
The invention es a kit for treating a condition mediated by Bruton’s tyrosine 15 kinase, comprising: a) a first pharmaceutical composition comprising a Formula I nd; and b) instructions for use.
The ion includes a Formula I compound for use as a ment, and for use in treating a disease or disorder selected from immune disorders, cancer, cardiovascular disease, viral infection, inflammation, metabolism/endocrine function disorders and neurological 20 disorders, and mediated by Bruton’s tyrosine kinase.
The invention includes use of a Formula I nd in the manufacture of a medicament for the treatment of immune disorders, cancer, cardiovascular disease, viral infection, inflammation, metabolism/endocrine function disorders and neurological disorders, and where the medicament mediates Bruton’s tyrosine kinase. 25 Also described herein are methods of making a Formula I compound.
BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows the preparation of 2-(4-(hydroxymethyl)(1-methyl(5-(4-(oxetan- 3-yl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 3 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 101 starting with 2,2,2-Trichloro (4,5,6,7-tetrahydro-1H-indolyl)ethanone 101a.
Figure 2 shows the preparation of 2-(4-(Hydroxymethyl)(1-methyl(5-(4- methylpiperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 5 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 102 starting with 1-Methyl[5-(4- methyl-piperazinyl)-pyridinylamino](4,4,5,5-tetramethyl-[1,3,2]dioxaborolanyl)- idinone 102a Figure 3 shows the preparation of 2-(3-(Hydroxymethyl)(1-methyl(5-(4- (oxetanyl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 10 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 103 starting with o chloronicotinaldehyde 103a Figure 4 shows the preparation of 2-(3-(Hydroxymethyl)(1-methyl(5-(4- (oxetanyl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 104 ng with 4-Bromo 15 chloronicotinaldehyde 104a Figure 5 shows the preparation of 4-Hydroxymethyl- 3-[1-methyl({5-[4-(oxetan yl)piperazinyl]pyridineyl}amino)oxo-1,6-dihydropyridinyl]{6-oxothia azatricyclo-[7.4.0.02,7]trideca-1(9),2(7)-dienyl}pyridine 105 starting with N-Methoxy-N- methyl-4,5,6,7-tetrahydrobenzo[b]thiophenecarboxamide 105a 20 Figure 6 shows the preparation of 4-Hydroxymethyl[1-methyl({5-[4-(oxetan erazinyl]pyridineyl}amino)oxo-1,6-dihydropyridinyl]{4,4-dimethyl oxothiaazatricyclo[6.4.0.02,6]dodeca-1(8), 2(6)-dienyl}pyridinecarbaldehyde 106 starting with 3,3-Dimethylcyclopentanone 106a Figure 7 shows the preparation of 10-[4-[1-Methyl({5-[4-(oxetanyl)piperazin 25 yl]pyridineyl}amino)oxo-1,6-dihydropyridinyl](hydroxymethyl)pyridinyl]-4,4- dimethyl-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienone 107 starting with hyl 3-(2-Chloro-4,4-dimethylcyclopentenyl)acrylate 107a Figure 8 shows the preparation of 2-(3-(Hydroxymethyl)(1-methyl(5-(4- (oxetanyl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 30 4,4-dimethyl-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienone 108 starting with 4- Chloro{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dien idinecarbaldehyde 108a.
Figure 9 shows the preparation of 2-(3-(Hydroxymethyl)(1-methyl(5-(4- (oxetanyl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 4 4,4-dimethylthiaazatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienone 109 starting with 4-Chloro{4,4-dimethyloxothia azatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dien yl}pyridinecarbaldehyde 109a.
Figure 10 shows the preparation of Hydroxymethyl)(1-methyl(6-(4- 5 methylpiperazinyl)pyridineylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 110 starting with 1-Methyl(6-(4- methylpiperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolan yl)pyridin-2(1H)-one 110a Figure 11 shows the preparation of 2-(3-(Hydroxymethyl)(1-methyl(5- 10 (morpholinecarbonyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 111 starting with (6-Aminopyridin yl)(morpholino)methanone 111a Figure 12 shows the preparation of 2-(4-(Hydroxymethyl)(1-methyl(5-(4- (oxetanyl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 15 3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin-1(2H)-one 112 starting with Methyl 5,6,7,8- Tetrahydroindolizinecarboxylate 112a Figure 13 shows the preparation of Hydroxymethyl)(1-methyl(5-methyl- 4,5,6,7-tetrahydropyrazolo [1,5-a]pyrazinylamino)oxo-1,6-dihydropyridinyl)pyridin- 2-yl)-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 113 starting with (3-Nitro-1H- 20 pyrazolyl)methanol 113a Figure 14 shows the preparation of (R)(4-(6-(4-(1,4-dimethyloxopiperazin yl)phenylamino)methyl oxo-4,5-dihydropyrazinyl)(hydroxymethyl)pyridinyl)- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 114 starting with (R)bromo(4- (1,4-dimethyloxopiperazinyl)phenylamino)methylpyrazin -2(1H)-one 114a 25 Figure 15 shows the preparation of 2-(3-(Hydroxymethyl)(1-methyl(5-methyl- 1H-pyrazolylamino)oxo-1,6-dihydropyridinyl)pyridinyl)-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-1(2H)-one 115 starting with omethyl(5-methyl- 1H-pyrazolylamino)pyridin-2(1H)-one 115a Figure 16 shows the preparation of oxymethyl[1-methyl({5-[4-(oxetan- 30 3-yl)piperazinyl]pyridineyl}amino)oxo-1,6-dihydropyridinyl]{6-oxothia- 4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridine 116 starting with 3-Bromo- 5-{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridine carbaldehyde 116a 5 Figure 17 shows the preparation of 2-(3-(hydroxymethyl)(1-methyl(5- lsulfonyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)-3,4,6,7,8,9- hexahydro-pyrazino[1,2-a]indol-1(2H)-one 117 starting with 5-(Methylthio)nitropyridine 117a 5 Figure 18 shows the preparation of 2-(4-(5-(5-Cyclopropyl-1H-pyrazolylamino) methyloxo-1,6-dihydropyridinyl)(hydroxymethyl)pyridinyl)-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-1(2H)-one 118 starting with tert-Butyl 5-Amino cyclopropyl-1H-pyrazolecarboxylate 118a Figure 19 shows the preparation of (S)(3-(Hydroxymethyl)(1-methyl(5-(2- 10 methyl(oxetanyl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridin yl)pyridinyl)-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 119 ng with (S)- (4-(1-Methyl(5-(2-methyl(oxetanyl)piperazinyl)pyridinylamino)oxo-1,6- dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridin- 3-yl)methyl e 119a 15 Figure 20 shows the preparation of 2-(4-(5-(5-(4-(2-Hydroxy methylpropyl)piperazinyl)pyridinylamino)methyloxo-1,6-dihydropyridinyl) (hydroxymethyl)pyridinyl)-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 120 starting with o(5-(4-(2-hydroxymethylpropyl)piperazinyl)pyridin ylamino)methylpyridin-2(1H)-one 120a 20 Figure 21 shows the preparation of 2-(3-(hydroxymethyl)(1-methyl(5-(4- (oxetanyl)piperazinyl)pyridineylamino)oxo-1,6-dihydropyridinyl)pyridin yl)-6,7,8,9-tetrahydropyrazino[1,2-a]indol-1(2H)-one 121 starting with 4-(1-Methyl(5-(4- (oxetanyl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)(1-oxo- 6,7,8,9-tetrahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 121a 25 Figure 22 shows the ation of 2-(4-(5-(5-((2S,5R)-2,5-Dimethyl(oxetan yl)piperazinyl)pyridinylamino)methyloxo-1,6-dihydropyridinyl) (hydroxymethyl)pyridinyl)-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 122 starting with (2R, 5S)-tert-Butyl 2,5-Dimethyl(6-nitropyridinyl)piperazine carboxylate 122a 30 Figure 23 shows the preparation of 2-(4-(5-(5-(4-(2-Hydroxyethyl)piperazin yl)pyridinylamino)methyloxo-1,6-dihydropyridinyl)(hydroxymethyl)pyridin yl)-3,4,6,7,8,9-hexahydro-pyrazino[1,2-a]indol-1(2H)-one 123 starting with (2- Bromoethoxy)(tert-butyl)dimethylsilane 123a 6 Figure 24 shows the preparation of 3-Hydroxymethyl- 4-[1-methyl(5-(4-(oxetan yl)piperazinyl)pyridineylamino)oxo-1,6-dihydropyridinyl]{6-oxothia-4,5- diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3- trienyl}pyridine 124 starting with 4-Chloro {6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridine 5 carbaldehyde 124a Figure 25 shows the preparation of 7,7-difluoro-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-1(2H)-one, useful for the preparation of 140, starting from ethyl 1H-pyrrole carboxylate.
Figure 26 shows the preparation of 5-(oxetanyl)-1H-pyrazolamine, useful for 10 the preparation of 266, starting from 3-nitro-1H-pyrazole.
ED DESCRIPTION OF EXEMPLARY EMBODIMENTS Reference will now be made in detail to certain embodiments of the ion, examples of which are illustrated in the accompanying structures and formulas. While the invention will be described in ction with the ated embodiments, it will be 15 tood that they are not intended to limit the invention to those ments. On the contrary, the invention is intended to cover all alternatives, modifications, and equivalents which may be included within the scope of the present invention as defined by the claims.
One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the ce of the t invention. The 20 present invention is in no way limited to the methods and materials described. In the event that one or more of the incorporated literature, patents, and similar materials differs from or contradicts this ation, including but not limited to defined terms, term usage, described techniques, or the like, this application ls. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of 25 ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the invention, suitable methods and materials are described below. All publications, patent applications, patents, and other nces mentioned herein are incorporated by reference in their entirety. The nomenclature used in this Application is based on IUPAC atic 30 nomenclature, unless indicated otherwise.
DEFINITIONS When indicating the number of substituents, the term “one or more” refers to the range from one substituent to the highest possible number of tution, i.e. replacement of 7 one hydrogen up to replacement of all hydrogens by substituents. The term “substituent” denotes an atom or a group of atoms replacing a hydrogen atom on the parent molecule. The term “substituted” denotes that a specified group bears one or more substituents. Where any group may carry multiple substituents and a variety of possible substituents is ed, the 5 substituents are independently selected and need not to be the same. The term “unsubstituted” means that the ied group bears no tuents. The term “optionally substituted” means that the specified group is unsubstituted or substituted by one or more substituents, ndently chosen from the group of possible substituents. When indicating the number of substituents, the term “one or more” means from one substituent to the highest le 10 number of substitution, i.e. replacement of one hydrogen up to replacement of all hydrogens by substituents.
The term “alkyl” as used herein refers to a saturated linear or branched-chain monovalent hydrocarbon radical of one to twelve carbon atoms (C1−C12), wherein the alkyl l may be optionally substituted independently with one or more substituents described 15 below. In another embodiment, an alkyl radical is one to eight carbon atoms (C1−C8), or one to six carbon atoms ). Examples of alkyl groups include, but are not limited to, methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl, -CH2CH2CH3), 2-propyl (i-Pr, i-propyl, -CH(CH3)2), 1-butyl (n-Bu, n-butyl, -CH2CH2CH2CH3), 2-methylpropyl (i- Bu, i-butyl, -CH2CH(CH3)2), 2-butyl (s-Bu, s-butyl, -CH(CH3)CH2CH3), 2-methylpropyl 20 (t-Bu, t-butyl, -C(CH3)3), yl (n-pentyl, -CH2CH2CH2CH2CH3), 2-pentyl (- CH(CH3)CH2CH2CH3), 3-pentyl H2CH3)2), ylbutyl (-C(CH3)2CH2CH3), 3- butyl (-CH(CH3)CH(CH3)2), 3-methylbutyl (-CH2CH2CH(CH3)2), 2-methyl butyl (-CH2CH(CH3)CH2CH3), 1-hexyl (-CH2CH2CH2CH2CH2CH3), 2-hexyl (- CH(CH3)CH2CH2CH2CH3), 3-hexyl H2CH3)(CH2CH2CH3)), 2-methylpentyl (- 25 C(CH3)2CH2CH2CH3), 3-methylpentyl (-CH(CH3)CH(CH3)CH2CH3), 4-methylpentyl (- CH(CH3)CH2CH(CH3)2), 3-methylpentyl (-C(CH3)(CH2CH3)2), 2-methylpentyl (- CH3)CH(CH3)2), 2,3-dimethylbutyl (-C(CH3)2CH(CH3)2), 3,3-dimethylbutyl (- CH(CH3)C(CH3)3, 1-heptyl, 1-octyl, and the like.
The term “alkylene” as used herein refers to a saturated linear or branched-chain 30 divalent hydrocarbon radical of one to twelve carbon atoms (C1−C12), wherein the alkylene radical may be optionally substituted independently with one or more substituents bed below. In another embodiment, an alkylene radical is one to eight carbon atoms (C1−C8), or 8 one to six carbon atoms (C1−C6). Examples of alkylene groups e, but are not limited to, methylene (-CH2-), ne (-CH2CH2-), propylene (-CH2CH2CH2-), and the like.
The term “alkenyl” refers to linear or branched-chain monovalent arbon radical of two to eight carbon atoms (C2−C8) with at least one site of unsaturation, i.e., a carbon- 5 carbon, sp2 double bond, wherein the alkenyl radical may be optionally tuted independently with one or more substituents described herein, and includes radicals having “cis” and “trans” orientations, or atively, “E” and “Z” ations. Examples include, but are not limited to, ethylenyl or vinyl (-CH=CH2), allyl (-CH2CH=CH2), and the like.
The term “alkenylene” refers to linear or branched-chain divalent hydrocarbon radical 10 of two to eight carbon atoms (C2−C8) with at least one site of unsaturation, i.e., a carboncarbon , sp2 double bond, wherein the lene radical may be optionally substituted substituted independently with one or more tuents described herein, and includes radicals having “cis” and ” orientations, or alternatively, “E” and “Z” ations.
Examples include, but are not limited to, ethylenylene or vinylene (-CH=CH-), allyl (- 15 CH-), and the like.
The term “alkynyl” refers to a linear or branched monovalent hydrocarbon radical of two to eight carbon atoms (C2−C8) with at least one site of unsaturation, i.e., a carbon-carbon, sp triple bond, wherein the alkynyl radical may be optionally substituted ndently with one or more substituents described herein. Examples include, but are not limited to, ethynyl 20 (-C≡CH), propynyl (propargyl, -CH2C≡CH), and the like.
The term “alkynylene” refers to a linear or ed divalent hydrocarbon radical of two to eight carbon atoms (C2−C8) with at least one site of unsaturation, i.e., a carbon-carbon, sp triple bond, wherein the alkynylene radical may be ally substituted independently with one or more substituents described herein. Examples include, but are not limited to, 25 ethynylene (-C≡C-), propynylene (propargylene, -CH2C≡C-), and the like.
The terms “carbocycle”, “carbocyclyl”, “carbocyclic ring” and “cycloalkyl” refer to a lent non-aromatic, saturated or partially unsaturated ring having 3 to 12 carbon atoms (C3−C12) as a monocyclic ring or 7 to 12 carbon atoms as a bicyclic ring. Bicyclic carbocycles having 7 to 12 atoms can be arranged, for example, as a bicyclo [4,5], [5,5], [5,6] 30 or [6,6] system, and bicyclic carbocycles having 9 or 10 ring atoms can be arranged as a bicyclo [5,6] or [6,6] system, or as bridged systems such as bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane and bicyclo[3.2.2]nonane. Spiro moieties are also included within the scope of this definition. Examples of monocyclic carbocycles include, but are not limited to, 9 cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopentenyl, 1-cyclopentenyl, opent enyl, cyclohexyl, 1-cyclohexenyl, 1-cyclohexenyl, 1-cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl, and the like.
Carbocyclyl groups are optionally substituted independently with one or more substituents 5 described herein.
“Aryl” means a monovalent aromatic hydrocarbon radical of 6-20 carbon atoms (C6− C20) derived by the l of one hydrogen atom from a single carbon atom of a parent aromatic ring system. Some aryl groups are ented in the exemplary structures as “Ar”.
Aryl includes bicyclic radicals comprising an aromatic ring fused to a ted, partially 10 unsaturated ring, or aromatic carbocyclic ring. Typical aryl groups include, but are not limited to, ls derived from benzene (phenyl), substituted benzenes, naphthalene, anthracene, biphenyl, l, indanyl, 1,2-dihydronaphthalene, 1,2,3,4-tetrahydronaphthyl, and the like. Aryl groups are optionally substituted independently with one or more substituents described herein. 15 “Arylene” means a divalent aromatic hydrocarbon radical of 6-20 carbon atoms (C6− C20) derived by the l of two hydrogen atom from a two carbon atoms of a parent ic ring system. Some arylene groups are represented in the ary structures as “Ar”. Arylene includes bicyclic radicals comprising an aromatic ring fused to a saturated, partially unsaturated ring, or aromatic yclic ring. Typical arylene groups include, but 20 are not limited to, radicals derived from benzene (phenylene), substituted benzenes, naphthalene, anthracene, biphenylene, indenylene, indanylene, 1,2-dihydronaphthalene, 4-tetrahydronaphthyl, and the like. Arylene groups are optionally substituted with one or more substituents described herein.
The terms ocycle,” “heterocyclyl” and “heterocyclic ring” are used 25 interchangeably herein and refer to a saturated or a partially unsaturated (i.e., having one or more double and/or triple bonds within the ring) carbocyclic radical of 3 to about 20 ring atoms in which at least one ring atom is a heteroatom ed from nitrogen, oxygen, phosphorus and sulfur, the remaining ring atoms being C, where one or more ring atoms is optionally substituted independently with one or more substituents described below. A 30 heterocycle may be a monocycle having 3 to 7 ring members (2 to 6 carbon atoms and 1 to 4 heteroatoms selected from N, O, P, and S) or a bicycle having 7 to 10 ring members (4 to 9 carbon atoms and 1 to 6 heteroatoms ed from N, O, P, and S), for example: a bicyclo [4,5], [5,5], [5,6], or [6,6] system. Heterocycles are described in Paquette, Leo A.; 10 “Principles of Modern cyclic Chemistry” (W.A. Benjamin, New York, 1968), particularly Chapters 1, 3, 4, 6, 7, and 9; “The Chemistry of Heterocyclic Compounds, A series of Monographs” (John Wiley & Sons, New York, 1950 to present), in particular Volumes 13, 14, 16, 19, and 28; and J. Am. Chem. Soc. (1960) 82:5566. “Heterocyclyl” also 5 includes radicals where heterocycle radicals are fused with a saturated, partially unsaturated ring, or aromatic carbocyclic or heterocyclic ring. Examples of heterocyclic rings e, but are not limited to, morpholinyl, piperidinyl, piperazinyl, piperazinylone, piperazinylone, pyrrolidinyl, rpholinyl, S-dioxothiomorpholinyl, azocanyl, azetidinyl, octahydropyrido[1,2-a]pyrazinyl, [1,4]diazepanyl, 10 pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, homopiperazinyl, azetidinyl, oxetanyl, thietanyl, peridinyl, oxepanyl, thiepanyl, inyl, diazepinyl, thiazepinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H- pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, linyl, dithianyl, dithiolanyl, 15 dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinylimidazolinyl, imidazolidinyl, 3- azabicyco[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, azabicyclo[2.2.2]hexanyl, 3H-indolyl quinolizinyl and N-pyridyl ureas. Spiro moieties are also included within the scope of this definition. Examples of a heterocyclic group wherein 2 ring atoms are substituted with oxo (=O) moieties are pyrimidinonyl and 1,1-dioxo-thiomorpholinyl. The heterocycle groups 20 herein are optionally substituted independently with one or more substituents described herein.
The term “heteroaryl” refers to a monovalent aromatic radical of 5-, 6-, or 7- ed rings, and includes fused ring systems (at least one of which is aromatic) of 5-20 atoms, containing one or more heteroatoms independently selected from en, oxygen, 25 and sulfur. Examples of heteroaryl groups are pyridinyl (including, for e, 2- hydroxypyridinyl), imidazolyl, imidazopyridinyl, dinyl (including, for example, 4- hydroxypyrimidinyl), pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, l, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, 30 indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, l, oxadiazolyl, lyl, thiadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, hiophenyl, hiazolyl, benzoxazolyl, olinyl, quinoxalinyl, yridinyl, and furopyridinyl.
Heteroaryl groups are optionally substituted independently with one or more substituents described herein. 11 The heterocycle or heteroaryl groups may be carbon (carbon-linked), or nitrogen (nitrogen-linked) bonded where such is possible. By way of example and not limitation, carbon bonded heterocycles or heteroaryls are bonded at position 2, 3, 4, 5, or 6 of a ne, position 3, 4, 5, or 6 of a pyridazine, position 2, 4, 5, or 6 of a pyrimidine, on 2, 3, 5, or 5 6 of a pyrazine, on 2, 3, 4, or 5 of a furan, tetrahydrofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole, position 2, 4, or 5 of an oxazole, imidazole or thiazole, position 3, 4, or 5 of an isoxazole, pyrazole, or isothiazole, position 2 or 3 of an aziridine, position 2, 3, or 4 of an azetidine, position 2, 3, 4, 5, 6, 7, or 8 of a quinoline or position 1, 3, 4, 5, 6, 7, or 8 of an isoquinoline. 10 By way of example and not limitation, nitrogen bonded heterocycles or heteroaryls are bonded at position 1 of an aziridine, azetidine, pyrrole, idine, oline, 3- pyrroline, ole, imidazolidine, 2-imidazoline, 3-imidazoline, pyrazole, pyrazoline, 2- pyrazoline, 3-pyrazoline, piperidine, piperazine, indole, indoline, 1H-indazole, position 2 of a isoindole, or isoindoline, position 4 of a morpholine, and position 9 of a carbazole, or β- 15 carboline.
The terms “treat” and “treatment” refer to therapeutic treatment, wherein the object is to slow down (lessen) an undesired physiological change or disorder, such as the development or spread of arthritis or cancer. For purposes of this invention, beneficial or d clinical results include, but are not limited to, alleviation of symptoms, diminishment 20 of extent of disease, ized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or , whether detectable or undetectable. “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment. Those in need of treatment include those with the condition or er. 25 The phrase “therapeutically effective amount” means an amount of a compound of the present invention that (i) treats the particular disease, condition, or disorder, (ii) attenuates, ameliorates, or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of the particular disease, ion, or disorder described herein. In the case of cancer, the therapeutically 30 effective amount of the drug may reduce the number of cancer cells; reduce the tumor size; inhibit (i.e., slow to some extent and preferably stop) cancer cell infiltration into eral organs; inhibit (i.e., slow to some extent and preferably stop) tumor metastasis; inhibit, to some extent, tumor ; and/or relieve to some extent one or more of the ms associated with the cancer. To the extent the drug may prevent growth and/or kill existing 12 cancer cells, it may be cytostatic and/or cytotoxic. For cancer therapy, efficacy can be measured, for e, by assessing the time to disease progression (TTP) and/or determining the response rate (RR).
"Inflammatory disorder" as used herein can refer to any disease, disorder, or 5 syndrome in which an excessive or unregulated inflammatory response leads to excessive inflammatory symptoms, host tissue damage, or loss of tissue function. "Inflammatory disorder" also refers to a pathological state mediated by influx of leukocytes and/or neutrophil axis. mmation" as used herein refers to a localized, protective response elicited by 10 injury or destruction of tissues, which serves to destroy, dilute, or wall off (sequester) both the injurious agent and the d tissue. Inflammation is notably ated with influx of leukocytes and/or neutrophil chemotaxis. Inflammation can result from infection with pathogenic organisms and viruses and from noninfectious means such as trauma or reperfusion following myocardial infarction or stroke, immune response to foreign antigen, 15 and autoimmune ses. Accordingly, inflammatory disorders amenable to treatment with a I compounds encompass disorders associated with reactions of the specific defense system as well as with reactions of the nonspecific defense system.
"Specific defense system" refers to the component of the immune system that reacts to the presence of ic antigens. Examples of inflammation resulting from a response of the 20 specific defense system include the classical response to foreign ns, autoimmune diseases, and delayed type hypersensitivity response ed by T-cells. Chronic inflammatory diseases, the ion of solid transplanted tissue and organs, e.g., kidney and bone marrow transplants, and graft versus host disease (GVHD), are further examples of inflammatory reactions of the specific defense system. 25 The term "nonspecific defense system" as used herein refers to matory disorders that are mediated by leukocytes that are incapable of immunological memory (e.g., granulocytes, and hages). Examples of inflammation that result, at least in part, from a reaction of the nonspecific e system include inflammation associated with conditions such as adult (acute) respiratory distress syndrome (ARDS) or multiple organ injury 30 syndromes; usion injury; acute glomerulonephritis; reactive arthritis; dermatoses with acute inflammatory components; acute purulent meningitis or other central nervous system matory disorders such as stroke; thermal injury; inflammatory bowel disease; granulocyte transfusion ated syndromes; and cytokine-induced toxicity. 13 "Autoimmune disease" as used herein refers to any group of disorders in which tissue injury is associated with humoral or cell-mediated responses to the body's own constituents.
"Allergic disease" as used herein refers to any symptoms, tissue damage, or loss of tissue function resulting from allergy. "Arthritic disease" as used herein refers to any disease 5 that is characterized by inflammatory lesions of the joints utable to a variety of etiologies. "Dermatitis" as used herein refers to any of a large family of diseases of the skin that are characterized by inflammation of the skin attributable to a variety of etiologies.
"Transplant ion" as used herein refers to any immune reaction directed against grafted tissue, such as organs or cells (e.g., bone marrow), characterized by a loss of function of the 10 grafted and surrounding tissues, pain, swelling, leukocytosis, and ocytopenia. The therapeutic methods of the present invention include methods for the treatment of disorders associated with inflammatory cell activation.
"Inflammatory cell activation" refers to the induction by a stimulus (including, but not limited to, cytokines, antigens or auto-antibodies) of a proliferative cellular response, the 15 production of soluble mediators ding but not d to cytokines, oxygen radicals, enzymes, noids, or vasoactive amines), or cell surface expression of new or sed numbers of mediators (including, but not limited to, major histocompatability antigens or cell adhesion molecules) in inflammatory cells (including but not limited to monocytes, macrophages, T lymphocytes, B lymphocytes, granulocytes (i.e., polymorphonuclear 20 leukocytes such as phils, basophils, and eosinophils), mast cells, dendritic cells, Langerhans cells, and endothelial cells). It will be appreciated by persons skilled in the art that the tion of one or a combination of these phenotypes in these cells can contribute to the initiation, perpetuation, or exacerbation of an inflammatory disorder.
The term "NSAID" is an acronym for "non-steroidal anti-inflammatory drug" and is a 25 therapeutic agent with analgesic, antipyretic (lowering an elevated body temperature and relieving pain without impairing consciousness) and, in higher doses, with anti-inflammatory effects (reducing inflammation). The term "non-steroidal" is used to guish these drugs from steroids, which (among a broad range of other s) have a r eicosanoiddepressing , anti-inflammatory action. As analgesics, NSAIDs are unusual in that they are 30 non-narcotic. NSAIDs include aspirin, ibuprofen, and naproxen. NSAIDs are usually indicated for the treatment of acute or chronic conditions where pain and mation are present. NSAIDs are generally indicated for the symptomatic relief of the following conditions: rheumatoid tis, osteoarthritis, inflammatory arthropathies (e.g. ankylosing spondylitis, psoriatic arthritis, Reiter's me, acute gout, dysmenorrhoea, metastatic bone 14 pain, headache and migraine, erative pain, mild-to-moderate pain due to inflammation and tissue injury, pyrexia, ileus, and renal colic. Most NSAIDs act as non-selective inhibitors of the enzyme cyclooxygenase, inhibiting both the cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) isoenzymes. Cyclooxygenase catalyzes the ion of 5 prostaglandins and thromboxane from arachidonic acid (itself derived from the cellular phospholipid r by olipase A2). Prostaglandins act (among other things) as messenger molecules in the process of inflammation. COX-2 inhibitors include celecoxib, etoricoxib, lumiracoxib, parecoxib, rofecoxib, rofecoxib, and valdecoxib.
The terms r” refers to or describe the physiological condition in mammals that 10 is typically characterized by lated cell growth. A “tumor” comprises one or more cancerous cells. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. More particular examples of such cancers include squamous cell cancer (e.g., epithelial squamous cell cancer), lung cancer including small- cell lung cancer, non-small cell lung cancer (“NSCLC”), adenocarcinoma of 15 the lung and squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular , gastric or stomach cancer including intestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, r cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal , prostate cancer, vulval cancer, thyroid 20 cancer, hepatic oma, anal carcinoma, penile carcinoma, as well as head and neck cancer.
"Hematological malignancies" (British spelling tological" malignancies) are the types of cancer that affect blood, bone marrow, and lymph nodes. As the three are intimately connected through the immune system, a disease affecting one of the three will often affect the others as well: although lymphoma is a disease of the lymph nodes, it often 25 spreads to the bone marrow, affecting the blood. Hematological malignancies are malignant neoplasms ("cancer"), and they are lly treated by specialists in hematology and/or oncology. In some centers "Hematology/oncology" is a single subspecialty of internal medicine while in others they are ered separate ons (there are also surgical and radiation oncologists). Not all hematological disorders are malignant ("cancerous"); these 30 other blood conditions may also be managed by a hematologist. Hematological malignancies may derive from either of the two major blood cell lineages: myeloid and lymphoid cell lines.
The myeloid cell line normally produces granulocytes, erythrocytes, thrombocytes, macrophages and mast cells; the lymphoid cell line produces B, T, NK and plasma cells. mas, cytic leukemias, and myeloma are from the lymphoid line, while acute 15 and chronic myelogenous leukemia, myelodysplastic syndromes and myeloproliferative diseases are myeloid in origin. Leukemias include Acute lymphoblastic leukemia (ALL), Acute myelogenous leukemia (AML), Chronic lymphocytic leukemia (CLL), Chronic enous leukemia (CML), Acute monocytic leukemia (AMOL) and small lymphocytic 5 ma (SLL). mas include Hodgkin's lymphomas (all four es) and Non- Hodgkin's lymphomas (all subtypes).
A “chemotherapeutic agent” is a chemical compound useful in the treatment of , regardless of mechanism of action. s of chemotherapeutic agents include, but are not limited to: alkylating agents, antimetabolites, spindle poison plant alkaloids, 10 cytotoxic/antitumor antibiotics, topoisomerase inhibitors, antibodies, photosensitizers, and kinase inhibitors. Chemotherapeutic agents include compounds used in “targeted therapy” and conventional chemotherapy. Examples of chemotherapeutic agents include: erlotinib (TARCEVA®, Genentech/OSI Pharm.), xel (TAXOTERE®, Sanofi-Aventis), 5-FU ouracil, 5-fluorouracil, CAS No. 8), abine (GEMZAR®, Lilly), PD- 15 0325901 (CAS No. 3912109, Pfizer), tin (cis-diamine, dichloroplatinum(II), CAS No. 156631), carboplatin (CAS No. 415754), paclitaxel ®, Bristol-Myers Squibb Oncology, Princeton, N.J.), trastuzumab (HERCEPTIN®, Genentech), temozolomide (4-methyloxo- 2,3,4,6,8-pentazabicyclo ] nona-2,7,9-triene- 9-carboxamide, CAS No. 856221, TEMODAR®, L®, ng Plough), tamoxifen -[4-(1,2- 20 diphenylbutenyl)phenoxy]-N,N-dimethylethanamine, NOLVADEX®, ISTUBAL®, VALODEX®), and doxorubicin (ADRIAMYCIN®), Akti-1/2, HPPD, and rapamycin.
More examples of chemotherapeutic agents include: oxaliplatin (ELOXATIN®, Sanofi), bortezomib (VELCADE®, Millennium Pharm.), sutent (SUNITINIB®, 8, Pfizer), letrozole (FEMARA®, Novartis), imatinib mesylate (GLEEVEC®, Novartis), XL- 25 518 (Mek inhibitor, Exelixis, WO 2007/044515), ARRY-886 (Mek inhibitor, AZD6244, Array BioPharma, Astra Zeneca), SF-1126 (PI3K inhibitor, Semafore Pharmaceuticals), BEZ-235 (PI3K inhibitor, Novartis), XL-147 (PI3K inhibitor, Exelixis), PTK787/ZK 222584 (Novartis), fulvestrant (FASLODEX®, AstraZeneca), leucovorin (folinic acid), rapamycin imus, RAPAMUNE®, Wyeth), lapatinib (TYKERB®, GSK572016, Glaxo Smith 30 Kline), lonafarnib (SARASAR™, SCH 66336, Schering Plough), sorafenib (NEXAVAR®, BAY43-9006, Bayer Labs), gefitinib A®, AstraZeneca), irinotecan (CAMPTOSAR®, CPT-11, Pfizer), tipifarnib (ZARNESTRA™, Johnson & Johnson), ABRAXANE™ (Cremophor-free), albumin-engineered nanoparticle formulations of paclitaxel (American Pharmaceutical Partners, Schaumberg, Il), vandetanib (rINN, ZD6474, ZACTIMA®, 16 AstraZeneca), chloranmbucil, AG1478, AG1571 (SU 5271; Sugen), temsirolimus (TORISEL®, , pazopanib (GlaxoSmithKline), canfosfamide (TELCYTA®, Telik), thiotepa and cyclosphosphamide (CYTOXAN®, NEOSAR®); alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, 5 and uredopa; nimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylomelamine; enins (especially bullatacin and bullatacinone); a camptothecin (including the synthetic analog topotecan); bryostatin; callystatin; CC-1065 ding its adozelesin, carzelesin and bizelesin synthetic analogs); cryptophycins (particularly 10 cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogs, KW-2189 and 1); eleutherobin; pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas such as 15 tine, chlorozotocin, fotemustine, lomustine, ine, and ranimnustine; otics such as the enediyne antibiotics (e.g., calicheamicin, eamicin gamma1I, calicheamicin omegaI1 (Angew Chem. Intl. Ed. Engl. (1994) 33:183-186); dynemicin, dynemicin A; bisphosphonates, such as clodronate; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomysins, 20 actinomycin, authramycin, azaserine, bleomycins, cactinomycin, cin, carminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, ooxo-L- norleucine, morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, nemorubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, 25 peplomycin, porfiromycin, cin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5- fluorouracil (5-FU); folic acid s such as denopterin, rexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as bine, azacitidine, 6-azauridine, carmofur, cytarabine, 30 dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, ostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide ide; aminolevulinic acid; eniluracil; ine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; uone; elfornithine; 17 elliptinium acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; nmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS 5 Natural Products, Eugene, OR); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2’,2”-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A and ine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (“Ara-C”); cyclophosphamide; thiotepa; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin 10 and latin; stine; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; vinorelbine BINE®); novantrone; teniposide; edatrexate; daunomycin; aminopterin; capecitabine (XELODA®, Roche); ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; and ceutically acceptable salts, acids and derivatives of any of the above. 15 Also ed in the definition of “chemotherapeutic agent” are: (i) anti-hormonal agents that act to regulate or inhibit hormone action on tumors such as strogens and selective estrogen or modulators (SERMs), including, for example, fen (including NOLVADEX®; tamoxifen citrate), raloxifene, droloxifene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, stone, and FARESTON® (toremifine citrate); (ii) 20 aromatase inhibitors that inhibit the enzyme aromatase, which tes estrogen production in the adrenal glands, such as, for example, 4(5)-imidazoles, aminoglutethimide, MEGASE® (megestrol acetate), AROMASIN® (exemestane; Pfizer), formestanie, fadrozole, RIVISOR® (vorozole), FEMARA® (letrozole; Novartis), and ARIMIDEX® (anastrozole; AstraZeneca); (iii) ndrogens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; as 25 well as troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); (iv) protein kinase inhibitors such as MEK inhibitors (WO 44515); (v) lipid kinase inhibitors; (vi) antisense oligonucleotides, particularly those which t expression of genes in signaling pathways implicated in aberrant cell eration, for example, PKC-alpha, Raf and H-Ras, such as oblimersen (GENASENSE®, Genta Inc.); (vii) ribozymes such as VEGF expression 30 inhibitors (e.g., ANGIOZYME®) and HER2 expression inhibitors; (viii) vaccines such as gene y vaccines, for example, ALLOVECTIN®, LEUVECTIN®, and VAXID®; PROLEUKIN® rIL-2; topoisomerase 1 inhibitors such as LURTOTECAN®; ABARELIX® rmRH; (ix) anti-angiogenic agents such as zumab (AVASTIN®, Genentech); and pharmaceutically acceptable salts, acids and derivatives of any of the above. 18 Also included in the definition of therapeutic agent” are therapeutic antibodies such as alemtuzumab (Campath), zumab (AVASTIN®, Genentech); cetuximab (ERBITUX®, Imclone); mumab (VECTIBIX®, Amgen), rituximab (RITUXAN®, Genentech/Biogen Idec), pertuzumab (OMNITARG™, 2C4, Genentech), trastuzumab 5 (HERCEPTIN®, Genentech), tositumomab (Bexxar, Corixia), and the antibody drug conjugate, gemtuzumab ozogamicin (MYLOTARG®, Wyeth).
Humanized monoclonal antibodies with therapeutic potential as chemotherapeutic agents in combination with the Btk inhibitors of the invention include: alemtuzumab, apolizumab, aselizumab, atlizumab, uzumab, bevacizumab, bivatuzumab mertansine, 10 cantuzumab mertansine, zumab, certolizumab pegol, cidfusituzumab, cidtuzumab, daclizumab, umab, efalizumab, epratuzumab, erlizumab, felvizumab, fontolizumab, gemtuzumab ozogamicin, inotuzumab ozogamicin, ipilimumab, labetuzumab, lintuzumab, mab, mepolizumab, motavizumab, motovizumab, natalizumab, nimotuzumab, nolovizumab, numavizumab, ocrelizumab, omalizumab, palivizumab, pascolizumab, 15 pecfusituzumab, pectuzumab, pertuzumab, pexelizumab, ralivizumab, ranibizumab, reslivizumab, reslizumab, resyvizumab, rovelizumab, ruplizumab, sibrotuzumab, siplizumab, sontuzumab, tacatuzumab tetraxetan, tadocizumab, talizumab, tefibazumab, tocilizumab, toralizumab, trastuzumab, zumab celmoleukin, tucusituzumab, umavizumab, urtoxazumab, and visilizumab. 20 A “metabolite” is a product produced h metabolism in the body of a specified compound or salt thereof. Metabolites of a compound may be fied using routine techniques known in the art and their activities determined using tests such as those described herein. Such products may result for example from the oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic cleavage, and the like, of 25 the administered compound. Accordingly, also described herein are metabolites of compounds of the invention, including compounds produced by a s comprising contacting a Formula I compound of this invention with a mammal for a period of time ient to yield a metabolic product thereof.
The term “package ” is used to refer to instructions customarily included in 30 commercial packages of therapeutic products, that contain information about the indications, usage, dosage, administration, contraindications and/or warnings ning the use of such therapeutic products. 19 The term “chiral” refers to les which have the property of erimposability of the mirror image r, while the term “achiral” refers to molecules which are superimposable on their mirror image partner.
The term “stereoisomers” refers to compounds which have identical chemical 5 tution, but differ with regard to the arrangement of the atoms or groups in space.
“Diastereomer” refers to a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical properties, e.g. melting points, boiling points, spectral properties, and reactivities.
Mixtures of diastereomers may separate under high resolution analytical procedures such as 10 electrophoresis and chromatography.
“Enantiomers” refer to two stereoisomers of a compound which are nonsuperimposable mirror images of one another.
Stereochemical definitions and conventions used herein generally follow S. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New 15 York; and Eliel, E. and Wilen, S., “Stereochemistry of Organic nds”, John Wiley & Sons, Inc., New York, 1994. The compounds of the ion may contain asymmetric or chiral centers, and therefore exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds of the invention, including but not limited to, diastereomers, omers and atropisomers, as well as mixtures thereof such as racemic 20 mixtures, form part of the present invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D and L, or R and S, are used to denote the absolute configuration of the molecule about its chiral center(s). The prefixes d and l or (+) and (-) are employed to designate the sign of rotation of polarized light by the 25 compound, with (-) or 1 g that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of one another. A specific stereoisomer may also be referred to as an omer, and a mixture of such isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture or a racemate, 30 which may occur where there has been no stereoselection or stereospecificity in a chemical reaction or process. The terms ic mixture” and ate” refer to an equimolar mixture of two enantiomeric species, devoid of optical activity. Enantiomers may be separated from a racemic e by a chiral separation method, such as supercritical fluid chromatography (SFC). Assignment of configuration at chiral centers in separated 20 enantiomers may be tentative, and depicted in Table 1 structures for illustrative purposes, while stereochemical ination awaits, such as x-ray crystallographic data.
The term “tautomer” or “tautomeric form” refers to ural isomers of different es which are interconvertible via a low energy barrier. For example, proton tautomers 5 (also known as prototropic tautomers) e interconversions via migration of a , such as keto-enol and enamine isomerizations. Valence tautomers include interconversions by reorganization of some of the bonding electrons.
The term aceutically able salts” denotes salts which are not biologically or otherwise undesirable. Pharmaceutically acceptable salts include both acid and base 10 addition salts. The phrase “pharmaceutically acceptable” indicates that the substance or composition must be compatible chemically and/or toxicologically, with the other ingredients comprising a formulation, and/or the mammal being d therewith.
The term “pharmaceutically acceptable acid addition salt” denotes those pharmaceutically acceptable salts formed with inorganic acids such as hydrochloric acid, 15 hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, and organic acids selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, ylic, and sulfonic classes of organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, ic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic 20 acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, embonic acid, phenylacetic acid, methanesulfonic acid “mesylate”, ethanesulfonic acid, p-toluenesulfonic acid, and salicyclic acid.
The term “pharmaceutically acceptable base addition salt” denotes those pharmaceutically acceptable salts formed with an organic or inorganic base. Examples of 25 acceptable inorganic bases include sodium, potassium, ammonium, m, ium, iron, zinc, copper, manganese, and um salts. Salts derived from pharmaceutically acceptable organic nontoxic bases includes salts of primary, secondary, and tertiary amines, substituted amines including naturally ing substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, 30 tripropylamine, ethanolamine, 2-diethylaminoethanol, trimethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, zine, piperidine, N- ethylpiperidine, and polyamine resins 21 A “solvate” refers to an association or x of one or more solvent molecules and a compound of the invention. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethylacetate, acetic acid, and ethanolamine. 5 The term “EC50” is the half maximal ive concentration” and s the plasma concentration of a particular compound required for obtaining 50% of the maximum of a particular effect in vivo.
The term “Ki” is the inhibition constant and denotes the absolute binding affinity of a particular inhibitor to a or. It is measured using competition binding assays and is equal 10 to the tration where the particular inhibitor would occupy 50% of the receptors if no competing ligand (e.g. a radioligand) was present. Ki values can be converted logarithmically to pKi values (-log Ki), in which higher values indicate exponentially greater potency.
The term “IC50” is the half maximal tory concentration and denotes the concentration of a particular compound required for obtaining 50% inhibition of a biological 15 process in vitro. IC50 values can be converted logarithmically to pIC50 values (-log IC50), in which higher values indicate exponentially greater potency. The IC50 value is not an absolute value but depends on experimental conditions e.g. concentrations employed, and can be converted to an absolute inhibition constant (Ki) using the Cheng-Prusoff equation em.
Pharmacol. (1973) 22:3099). Other percent inhibition parameters, such as IC70, IC90, etc., 20 may be calculated.
The terms “compound of this ion,” and “compounds of the present invention” and “compounds of Formula I” include compounds of Formulas I and stereoisomers, geometric isomers, ers, solvates, and pharmaceutically acceptable salts thereof.
Any a or structure given herein, including Formula I compounds, is also 25 intended to represent hydrates, solvates, and polymorphs of such compounds, and mixtures thereof.
Any formula or structure given herein, including Formula I compounds, is also intended to represent unlabeled forms as well as isotopically labeled forms of the compounds. ically labeled compounds have structures depicted by the as given herein except 30 that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into compounds of the ion include es of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, and chlorine, such as, but not limited to 2H (deuterium, D), 3H (tritium), 11C, 13C, 14C, 15N, 18F, 31P, 32P, 35S, 36Cl, and 125I. s ically labeled compounds of the present invention, 22 for example those into which ctive isotopes such as 3H, 13C, and 14C are orated.
Such isotopically labelled nds may be useful in metabolic studies, reaction kinetic studies, detection or imaging ques, such as positron emission tomography (PET) or single-photon emission computed aphy (SPECT) including drug or substrate tissue 5 distribution assays, or in radioactive treatment of patients. Deuterium labelled or substituted therapeutic compounds of the invention may have improved DMPK (drug metabolism and pharmacokinetics) properties, relating to distribution, metabolism, and excretion (ADME).
Substitution with heavier isotopes such as deuterium may afford certain therapeutic advantages resulting from greater lic stability, for e increased in vivo half-life 10 or d dosage requirements. An 18F labeled compound may be useful for PET or SPECT studies. Isotopically labeled nds of this invention and prodrugs thereof can generally be prepared by carrying out the ures disclosed in the schemes or in the examples and preparations described below by substituting a readily available isotopically d reagent for a non-isotopically labeled reagent. Further, substitution with heavier isotopes, 15 particularly deuterium (i.e., 2H or D) may afford certain therapeutic ages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements or an improvement in therapeutic index. It is understood that deuterium in this context is regarded as a substituent in the compound of the formula (I). The concentration of such a heavier isotope, specifically deuterium, may be defined by an isotopic enrichment 20 factor. In the compounds of this invention any atom not ically designated as a particular isotope is meant to represent any stable isotope of that atom. Unless otherwise stated, when a position is designated specifically as "H" or "hydrogen", the position is understood to have hydrogen at its natural abundance isotopic composition. Accordingly, in the compounds of this ion any atom specifically ated as a deuterium (D) is meant to represent 25 deuterium.
HETEROARYL PYRIDONE AND AZA-PYRIDONE COMPOUNDS The present invention provides heteroaryl pyridone and aza-pyridone nds of Formula I, including Formulas Ia-Ii, and pharmaceutical formulations f, which are potentially useful in the treatment of diseases, conditions and/or disorders modulated by Btk 30 kinase: 23 I including stereoisomers, tautomers, or pharmaceutically acceptable salts thereof, wherein: X1 is CR1 or N; 5 X2 is CR2 or N; X3 is CR3 or N; where one or two of X1, X2, and X3 are N; R1, R2 and R3 are independently selected from H, F, Cl, −NH2, −NHCH3, −N(CH3)2, − OH, −OCH3, −OCH2CH3, −OCH2CH2OH, and C1−C3 alkyl; 10 R4 is selected from H, F, Cl, CN, −CH2OH, −CH(CH3)OH, −C(CH3)2OH, − CH(CF3)OH, −CH2F, −CHF2, −CH2CHF2, −CF3, −C(O)NH2, −C(O)NHCH3, (CH3)2, −NH2, −NHCH3, −N(CH3)2, −NHC(O)CH3, −OH, −OCH3, H3, H2OH, cyclopropyl, cyclopropylmethyl, 1-hydroxycyclopropyl, imidazolyl, pyrazolyl, 3-hydroxyoxetanyl , oxetanyl, and inyl; 15 R5 is optionally substituted C6−C20 aryl, C3−C12 carbocyclyl, C2−C20 heterocyclyl, C1−C20 heteroaryl, −(C6−C20 aryl)−(C2−C20 heterocyclyl), −(C1−C20 heteroaryl)−(C2−C20 heterocyclyl), −(C1−C20 heteroaryl)−(C2−C20 heterocyclyl)−(C2−C20 cyclyl), 20 heteroaryl)−(C2−C20 heterocyclyl)−(C1−C6 alkyl), −(C1−C20 heteroaryl)−(C1−C6 alkyl), −(C2− C20 cyclyl)−(C1−C6 alkyl), −(C2−C20 heterocyclyl)−(C3−C12 carbocyclyl), −(C1−C20 20 heteroaryl)−(C3−C12 carbocyclyl), or −(C1−C20 heteroaryl)−C(=O)−(C2−C20 heterocyclyl); R6 is H, F, −CH3, −CH2CH3, −CH2CH2OH, −NH2, or −OH; R7 is selected from the structures: 24 N N N N N N N N N N N N F O O F O O N N N N N N N S N F O O F O O N N N N N N N N N N N O F O O O N N N N N S N N F F O O O O 5 25 where the wavy line indicates the site of attachment; and Y1 and Y2 are independently selected from CH and N, where Y1 and Y2 are not each N; where alkyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl are optionally substituted 5 with one or more groups independently selected from F, Cl, Br, I, −CN, −CH3, −CH2CH3, − CH(CH3)2, −CH2CH(CH3)2, −CH2OH, H3, −CH2CH2OH, −C(CH3)2OH, − CH(OH)CH(CH3)2, −C(CH3)2CH2OH, −CH2CH2SO2CH3, −CH2OP(O)(OH)2, −CH2F, −CHF2, −CF3, −CH2CF3, −CH2CHF2, −CH(CH3)CN, −C(CH3)2CN, −CH2CN, −CO2H, , − CO2CH3, −CO2C(CH3)3, −COCH(OH)CH3, −CONH2, −CONHCH3, −CON(CH3)2, − 10 C(CH3)2CONH2, −NH2, −NHCH3, −N(CH3)2, −NHCOCH3, )COCH3, )2CH3, −N(CH3)C(CH3)2CONH2, −N(CH3)CH2CH2S(O)2CH3, −NO2, =O, −OH, −OCH3, − OCH2CH3, −OCH2CH2OCH3, −OCH2CH2OH, −OCH2CH2N(CH3)2, −OP(O)(OH)2, − (CH3)2, −SCH3, CH3, −S(O)3H, cyclopropyl, oxetanyl, azetidinyl, 1- methylazetidinyl)oxy, N-methyl-N-oxetanylamino, azetidinylmethyl, and 15 morpholino.
Exemplary embodiments of Formula I compounds include compounds of Formulas Ia-c: Exemplary embodiments of a I compounds also include compounds of 20 Formulas Id-i: 26 R5 R5 R5 NH NH NH O O O R4 Y2 R4 Y2 R4 Y2 R7 N R7 N R7 N Y1 R6 Y1 R6 Y1 R6 N N R3 R1 N R3 R1 R2 Id Ie If R2 R5 R5 R5 NH NH NH O O O R4 Y2 R4 Y2 R4 Y2 R7 N R7 N R7 N Y1 R6 Y1 R6 Y1 R6 N N N N N R3 R1 N R2 Ig Ih Ii Exemplary embodiments of Formula I compounds include n X1 is N, X1 is N, X1 is N, X1 and X3 are N, X1 and X2 are N, or X2 and X3 are N, as shown in Formulas Ic-Ii.
Exemplary embodiments of Formula I compounds include wherein R5 is optionally 5 substituted C1−C20 heteroaryl selected from pyrazolyl, pyridinyl, pyrimidinyl, 5-methyl- 4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinyl, 5-acetyl-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinyl, 6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazinyl, and 1-methyl(5-(4- methylpiperazinyl)pyridinyl.
Exemplary embodiments of Formula I compounds include wherein R5 is −(C1−C20 10 heteroaryl)−(C2−C20 cyclyl) where heteroaryl is optionally substituted pyridinyl and heterocyclyl is optionally tuted piperazinyl.
Exemplary embodiments of Formula I compounds include wherein R5 is , optionally substituted with one or more groups selected from F, Cl, −CH3, −S(O)2CH3, ropyl, azetidinyl, oxetanyl, and morpholino. 15 Exemplary ments of Formula I compounds include wherein R5 is selected from the structures: 27 O O O O N N N N N N N N N N N N HO HO O O N N N N N N N N N N N N O O N N O N N N N N N N N N 28 O O O O N N N O N S N O N N O H H N N N S O S O O O N N N N N S O S O S 29 where the wavy line indicates the site of attachment.
Exemplary embodiments of Formula I compounds include n R5 is: R8 N N N 5 where R8 is selected from H, −CH3, −CH2OCH3, −CH2CH3, −CH(CH3)2, − CH2CH2OH, −CH2CH2OCH3, −CH2F, −CHF2, −CF3, −CH2CF3, −CH2CHF2, −CH(CH3)CN, 30 −C(CH3)2CN, −CH2CN, −C(O)CH3, −C(O)CH2CH3, −C(O)CH(CH3)2, −NH2, −NHCH3, − N(CH3)2, −OH, −OCH3, −OCH2CH3, −OCH2CH2OH, cyclopropyl, and oxetanyl.
Exemplary embodiments of Formula I compounds include wherein R6 is CH3.
Exemplary embodiments of Formula I compounds include wherein Y1 is CH and Y2 5 is N, Y1 is N and Y2 is CH, Y1 and Y2 are each CH, or Y1 and Y2 are each CH and R6 is CH3.
Exemplary embodiments of Formula I nds include the compounds in Tables 1 and 2.
The a I compounds of the invention may contain asymmetric or chiral centers, and therefore exist in different stereoisomeric forms. It is intended that all stereoisomeric 10 forms of the compounds of the ion, including but not limited to, diastereomers, enantiomers and somers, as well as mixtures thereof such as racemic mixtures, form part of the present ion.
In addition, the present invention embraces all diastereomers, including cis-trans (geometric) and conformational isomers. For example, if a Formula I nd orates 15 a double bond or a fused ring, the cis- and trans-forms, as well as mixtures thereof, are embraced within the scope of the invention.
In the structures shown herein, where the stereochemistry of any particular chiral atom is not specified, then all stereoisomers are contemplated and included as the compounds of the invention. Where stereochemistry is ied by a solid wedge or dashed line 20 representing a particular uration, then that stereoisomer is so specified and defined.
The compounds of the present invention may exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and it is intended that the invention embrace both solvated and unsolvated forms.
The compounds of the present invention may also exist in different eric forms, 25 and all such forms are embraced within the scope of the invention. The term “tautomer” or “tautomeric form” refers to structural s of different energies which are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via ion of a proton, such as keto-enol and imineenamine isomerizations. Valence tautomers include interconversions by reorganization of 30 some of the bonding electrons.
BIOLOGICAL TION The relative efficacies of Formula I compounds as inhibitors of an enzyme activity (or other biological activity) can be established by determining the concentrations at which each 31 compound inhibits the activity to a predefined extent and then comparing the results.
Typically, the preferred determination is the concentration that inhibits 50% of the activity in a mical assay, i.e., the 50% inhibitory concentration or “IC50”. Determination of IC50 values can be accomplished using conventional techniques known in the art. In general, an 5 IC50 can be determined by measuring the activity of a given enzyme in the presence of a range of concentrations of the tor under study. The experimentally obtained values of enzyme activity then are plotted against the inhibitor concentrations used. The concentration of the inhibitor that shows 50% enzyme activity (as compared to the activity in the absence of any inhibitor) is taken as the IC50 value. Analogously, other inhibitory concentrations can be 10 defined h appropriate determinations of activity. For example, in some gs it can be desirable to establish a 90% inhibitory concentration, i.e., IC90, etc.
Formula I compounds were tested by a standard biochemical Btk Kinase Assay (Example 901).
A general procedure for a standard cellular Btk Kinase Assay that can be used to test 15 Formula I compounds is a Ramos Cell Btk Assay (Example 902).
A standard cellular B-cell proliferation assay can be used to test Formula I compounds with B-cells purified from spleen of Balb/c mice le 903).
A standard T cell proliferation assay can be used to test Formula I compounds with T- cells purified from spleen of Balb/c mice (Example 904). 20 A CD86 Inhibition assay can be conducted on Formula I compounds for the inhibition of B cell activity using total mouse splenocytes purified from spleens of 8-16 week old Balb/c mice (Example 905).
A B-ALL Cell Survival Assay can be conducted on Formula I compounds to measure the number of viable B-ALL cells in culture (Example 906). 25 A CD69 Whole Blood Assay can be conducted on Formula I compounds to determine the y of nds to t the production of CD69 by B lymphocytes in human whole blood activated by crosslinking surface IgM with goat F(ab’)2 anti-human IgM le 907). CD69 is a type II C-type lectin ed in lymphocyte ion and cytokine secretion. CD69 expression represents one of the earliest available indicators of 30 leukocyte activation and its rapid induction occurs through transcriptional activation (Vazquez et al (2009) Jour. of Immunology Published October 19, 2009, .4049/jimmunol.0900839). Concentration-dependent inhibition of antigen receptor stimulation by selective Btk inhibitors s cell surface expression of the lymphocyte activation marker CD69 (Honigberg et al (2010) Proc. Natl. Acad. Sci. 107(29):13075- 32 13080). Thus, CD69 inhibition by selective Btk inhibitors may be correlated with therapeutic efficacy of certain B-cell disorders. The CD69 Hu Blood FACS IC70 values are displayed for exemplary Formula I compounds in Tables 1 and 2.
The cytotoxic or cytostatic activity of Formula I exemplary compounds can be 5 measured by: establishing a proliferating mammalian tumor cell line in a cell culture medium, adding a Formula I compound, culturing the cells for a period from about 6 hours to about 5 days; and measuring cell viability (Example 908). Cell-based in vitro assays are used to measure viability, i.e. proliferation (IC50), cytotoxicity (EC50), and ion of apoptosis (caspase activation) and may be useful in predicting clinical efficacy against hematological 10 ancies and solid tumors.
The in vitro y of the combinations of Formula I compounds with chemotherapeutic agents can be measured by the cell eration assay of Example 908; the CellTiter-Glo® scent Cell Viability Assay, commercially available from Promega Corp., Madison, WI. This homogeneous assay method is based on the recombinant 15 expression of Coleoptera luciferase (US 5583024; US 5674713; US 5700670) and determines the number of viable cells in culture based on quantitation of the ATP present, an indicator of lically active cells (Crouch et al (1993) J. Immunol. Meth. 160:81-88; US 6602677). The ter-Glo® Assay was conducted in 96 or 384 well format, making it amenable to automated hroughput screening (HTS) (Cree et al (1995) ncer 20 Drugs 6:398-404). The homogeneous assay ure involves adding the single reagent (CellTiter-Glo® Reagent) directly to cells ed in serum-supplemented medium. Cell washing, removal of medium and multiple pipetting steps are not required. The system detects as few as 15 cells/well in a 384-well format in 10 minutes after adding reagent and mixing. 25 The neous "add-mix-measure" format results in cell lysis and generation of a scent signal proportional to the amount of ATP present. The amount of ATP is directly proportional to the number of cells present in culture. The CellTiter-Glo® Assay generates a "glow-type" luminescent signal, produced by the luciferase reaction, which has a half-life generally greater than five hours, depending on cell type and medium used. Viable cells are 30 reflected in relative luminescence units (RLU). The substrate, Beetle Luciferin, is oxidatively decarboxylated by recombinant firefly luciferase with concomitant conversion of ATP to AMP and generation of s. The extended half-life eliminates the need to use reagent ors and provides flexibility for continuous or batch mode processing of multiple plates. This cell proliferation assay can be used with various multiwell formats, e.g. 96 or 33 384 well format. Data can be recorded by luminometer or CCD camera g device. The luminescence output is ted as relative light units (RLU), measured over time.
The anti-proliferative efficacy of Formula I exemplary compounds and combinations with chemotherapeutic agents are measured by the CellTiter-Glo® Assay (Example 908) 5 against certain hematological tumor cell lines. EC50 values are established for the tested compounds and combinations.
Exemplary Formula I nds in Tables 1 and 2 were made, characterized, and tested for inhibition of Btk according to the methods of this invention, and have the following ures and corresponding names (ChemDraw Ultra, Version 9.0.1, and ChemBioDraw, 10 Version 11.0, CambridgeSoft Corp., Cambridge MA). Where more than one name is associated with a Formula I compound or intermediate, the chemical structure shall define the compound.
Table 1. 15 No. Structure Name Mol CD69 Weight Hu Blood FACS IC70 (µM) 101 2-{4-Hydroxymethyl-1'- 636.74 0.132 methyl-5'-[5-(4-oxetanylpiperazinyl )-pyridin ylamino]-6'-oxo-1',6'-dihydro- [3,3']bipyridinylyl}- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 102 2-{4-Hydroxymethyl-1'- 594.71 0.132 -5'-[5-(4-methylpiperazinyl )-pyridin ylamino]-6'-oxo-1',6'-dihydro- [3,3']bipyridinylyl}- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 34 103 2-(3-(Hydroxymethyl)(1- 636.74 0.0776 methyl(5-(4-(oxetan yl)piperazinyl)pyridin o)oxo-1,6- dihydropyridinyl)pyridin yl)-3,4,6,7,8,9- hexahydropyrazino[1,2- a]indol-1(2H)-one 104 2-(3-(Hydroxymethyl)(1- 636.74 0.793 methyl(5-(4-(oxetan yl)piperazinyl)pyridin ylamino)oxo-1,6- dihydropyridinyl)pyridin yl)-3,4,6,7,8,9- hexahydropyrazino[1,2- a]indol-1(2H)-one 105 2-{4-Hydroxymethyl-1'- 653.79 0.0654 methyl-5'-[5-(4-oxetanylpiperazinyl )-pyridin ylamino]-6'-oxo-1',6'-dihydro- bipyridinylyl}- 3,4,5,6,7,8-hexahydro-2H- benzo[4,5]thieno[2,3- c]pyridinone 106 ydroxymethyl-1'- 667.82 0.0576 methyl-5'-[5-(4-oxetanylpiperazinyl )-pyridin ylamino]-6'-oxo-1',6'-dihydro- [3,3']bipyridinylyl}-2,2- yl-2,3,5,6-tetrahydro- 1H,4Hthiaazacyclopenta [a]indenone 107 2-{4-Hydroxymethyl-1'- 650.77 0.0216 methyl-5'-[5-(4-oxetanylpiperazinyl )-pyridin ylamino]-6'-oxo-1',6'-dihydro- [3,3']bipyridinylyl}-7,7- dimethyl-3,4,7,8-tetrahydro- 2H,6H- cyclopenta[4,5]pyrrolo[1,2- a]pyrazinone 35 108 2-{3'-Hydroxymethyl 650.77 0.0319 methyl[5-(4-oxetanylpiperazinyl )-pyridin ylamino]oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}-7,7- dimethyl-3,4,7,8-tetrahydro- 2H,6H- cyclopenta[4,5]pyrrolo[1,2- a]pyrazinone 109 6-{3'-Hydroxymethyl 667.82 0.0501 methyl[5-(4-oxetanylpiperazinyl )-pyridin ylamino]oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}-2,2- dimethyl-2,3,5,6-tetrahydro- 1H,4Hthiaazacyclopenta [a]indenone 110 2-{3'-Hydroxymethyl 594.71 2.7 methyl[6-(4-methylpiperazinyl din o]oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- 3,4,6,7,8,9-hexahydro-2H- no[1,2-a]indolone 111 2-{3'-Hydroxymethyl 609.68 0.131 methyl[5-(morpholine carbonyl)-pyridinylamino]- 6-oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 112 2-{4-Hydroxymethyl-1'- 636.74 0.492 methyl-5'-[5-(4-oxetanylpiperazinyl )-pyridin ylamino]-6'-oxo-1',6'-dihydro- [3,3']bipyridinylyl}- 2,3,5,6,7,8-hexahydro-4H- 2,4b-diaza-fluorenone 36 113 2-[3'-Hydroxymethyl 554.64 0.0625 N methyl(5-methyl-4,5,6,7- tetrahydro-pyrazolo[1,5- N a]pyrazinylamino)oxo- N NH 1,6-dihydro-[3,4']bipyridinyl- HO O 2'-yl]-3,4,6,7,8,9-hexahydro- N azino[1,2-a]indolone N N O N 114 2-(4-{6-[4-((R)-1,4-Dimethyl- 622.72 0.0802 3-oxo-piperazinyl)- phenylamino]methyl oxo-4,5-dihydro-pyrazin yl}hydroxymethyl-pyridin- 2-yl)-3,4,6,7,8,9-hexahydro- azino[1,2-a]indolone 115 2-[3'-Hydroxymethyl 499.56 0.286 methyl(5-methyl-1H- pyrazolylamino)oxo-1,6- dihydro-[3,4']bipyridinyl-2'- yl]-3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 116 3-{4-Hydroxymethyl-1'- 652.77 0.377 methyl-5'-[5-(4-oxetanylpiperazinyl )-pyridin ylamino]-6'-oxo-1',6'-dihydro- bipyridinylyl}-6,7,8,9- tetrahydro-3H- benzo[4,5]thieno[2,3- d]pyridazinone 117 2-[3'-Hydroxymethyl(5- 574.65 0.396 methanesulfonyl-pyridin ylamino)methyloxo-1,6- dihydro-[3,4']bipyridinyl-2'- yl]-3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 37 118 2-[5-(5-Cyclopropyl-1H- 525.60 0.608 pyrazolylamino)-3'- hydroxymethylmethyl oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl]- 7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 119 2-{3'-Hydroxymethyl 650.77 0.0356 methyl[5-((S)methyl oxetanyl-piperazinyl)- pyridinylamino]oxo-1,6- dihydro-[3,4']bipyridinyl-2'- yl}-3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 120 2-(3'-Hydroxymethyl{5-[4- 652.79 0.283 (2-hydroxymethyl-propyl)- piperazinyl]-pyridin ylamino}methyloxo-1,6- dihydro-[3,4']bipyridinyl-2'- yl)-3,4,6,7,8,9-hexahydro-2H- no[1,2-a]indolone 121 2-{3'-Hydroxymethyl 634.73 0.0323 methyl[5-(4-oxetanylpiperazinyl )-pyridin ylamino]oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- 6,7,8,9-tetrahydro-2H- pyrazino[1,2-a]indolone 122 2-{5-[5-((2S,5R)-2,5- 664.80 0.0127 Dimethyloxetanylpiperazinyl )-pyridin ylamino]-3'-hydroxymethyl methyloxo-1,6-dihydro- bipyridinyl-2'-yl}- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 38 123 2-(5-{5-[4-(2-Hydroxy-ethyl)- 624.73 0.0331 piperazinyl]-pyridin ylamino}-3'-hydroxymethyl methyloxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl)- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 124 3-{3'-Hydroxymethyl 652.77 0.0362 methyl[5-(4-oxetanylpiperazinyl din ylamino]oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- 6,7,8,9-tetrahydro-3H- benzo[4,5]thieno[2,3- d]pyridazinone 125 2-[3'-Hydroxymethyl 596.68 0.0873 methyl(5-oxetanyl- 4,5,6,7-tetrahydropyrazolo [1,5-a]pyrazin ylamino)oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl]- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 126 2-{4-Hydroxymethyl-1'- 634.73 0.138 methyl-5'-[5-(4-oxetanylpiperazinyl )-pyridin ylamino]-6'-oxo-1',6'-dihydro- [3,3']bipyridinylyl}-6,7,8,9- ydro-2H-pyrazino[1,2- a]indolone 127 2-[3'-Hydroxymethyl 580.68 0.141 methyloxo(5-piperazin- 1-yl-pyridinylamino)-1,6- o-[3,4']bipyridinyl-2'- yl]-3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 39 128 2-[5-(5-Cyclopropyl-4,5,6,7- 580.68 0.0918 tetrahydro-pyrazolo[1,5- a]pyrazinylamino)-3'- hydroxymethylmethyl oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl]- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 129 2-[5-(6,7-Dihydro-4H- 541.60 0.0917 pyrazolo[5,1-c][1,4]oxazin ylamino)-3'-hydroxymethyl methyloxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl]- 3,4,6,7,8,9-hexahydro-2H- no[1,2-a]indolone 130 2-{3'-Hydroxymethyl 664.80 0.012 methyl[5-((S)methyl yl-piperazinyl)- pyridinylamino]oxo-1,6- dihydro-[3,4']bipyridinyl-2'- yl}-7,7-dimethyl-3,4,7,8- tetrahydro-2H,6H- cyclopenta[4,5]pyrrolo[1,2- a]pyrazinone 131 2-{5-[5-((S)Ethyloxetan- 664.80 0.0155 3-yl-piperazinyl)-pyridin ylamino]-3'-hydroxymethyl methyloxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 132 2-{4-[5-(6,7-Dihydro-4H- 542.59 0.263 lo[5,1-c][1,4]oxazin ylamino)methyloxo-1,6- dihydro-pyridazinyl] hydroxymethyl-pyridinyl}- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 40 133 2-{3-Hydroxymethyl[1- 555.63 0.227 methyl(5-methyl-4,5,6,7- tetrahydro-pyrazolo[1,5- a]pyrazinylamino)oxo- 1,6-dihydro-pyridazinyl]- nyl}-3,4,6,7,8,9- hexahydro-2H-pyrazino[1,2- a]indolone 134 10-Fluoro{3'- 654.73 0.0944 hydroxymethylmethyl[5- (4-oxetanyl-piperazin yl)-pyridinylamino]oxo- 1,6-dihydro-[3,4']bipyridinyl- 2'-yl}-3,4,6,7,8,9-hexahydro- 2H-pyrazino[1,2-a]indolone 135 10-Fluoro[3'- 572.63 0.107 hydroxymethylmethyl(5- methyl-4,5,6,7-tetrahydropyrazolo [1,5-a]pyrazin ylamino)oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl]- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 136 10-Fluoro{3'- 668.76 0.030 ymethylmethyl[5- ((S)methyloxetanylpiperazinyl )-pyridin ylamino]oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 137 Hydroxymethyl 650.77 0.0646 methyl[5-((R)methyl oxetanyl-piperazinyl)- pyridinylamino]oxo-1,6- dihydro-[3,4']bipyridinyl-2'- yl}-3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 41 138 2-[4-Hydroxymethyl-1'- 554.64 0.353 methyl-5'-(5-methyl-4,5,6,7- tetrahydro-pyrazolo[1,5- a]pyrazinylamino)-6'-oxo- 1',6'-dihydro-[3,3']bipyridinyl- 5-yl]-3,4,6,7,8,9-hexahydro- 2H-pyrazino[1,2-a]indolone 139 2-{3'-Hydroxymethyl 636.74 0.326 methyl[5-(4-oxetanylpiperazinyl )-pyridin ylamino]oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- 2,3,5,6,7,8-hexahydro-4H- 2,4b-diaza-fluorenone 140 7,7-Difluoro{3'- 686.75 0.308 hydroxymethylmethyl[5- ((S)methyloxetanylpiperazinyl )-pyridin o]oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 141 2-[3'-Hydroxymethyl 568.67 0.0266 methyl(5-methyl-4,5,6,7- tetrahydro-pyrazolo[1,5- a]pyrazinylamino)oxo- hydro-[3,4']bipyridinyl- 2'-yl]-7,7-dimethyl-3,4,7,8- tetrahydro-2H,6H- cyclopenta[4,5]pyrrolo[1,2- a]pyrazinone 142 2-[3'-Hydroxymethyl 497.55 2.1 oxo(pyrimidin o)-1,6-dihydro- [3,4']bipyridinyl-2'-yl]- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 143 N 6-[3'-Hydroxymethyl 528.63 0.0309 methyloxo(pyrimidin N NH ylamino)-1,6-dihydro- HO O [3,4']bipyridinyl-2'-yl]-2,2- dimethyl-2,3,5,6-tetrahydro- S N N 1H,4Hthiaaza- O N cyclopenta[a]indenone 42 144 2-[3'-Hydroxymethyl 511.58 0.106 methyloxo(pyrimidin ylamino)-1,6-dihydro- [3,4']bipyridinyl-2'-yl]-7,7- dimethyl-3,4,7,8-tetrahydro- 2H,6H- cyclopenta[4,5]pyrrolo[1,2- zinone 145 6-{3'-Hydroxymethyl 681.85 0.0147 methyl[5-((S)methyl oxetanyl-piperazinyl)- pyridinylamino]oxo-1,6- o-[3,4']bipyridinyl-2'- yl}-2,2-dimethyl-2,3,5,6- tetrahydro-1H,4Hthia aza-cyclopenta[a]indenone 146 10-Fluoro[3'- 515.54 0.0856 hydroxymethylmethyl (pyrimidinylamino)- 1,6-dihydro-[3,4']bipyridinyl- 2'-yl]-3,4,6,7,8,9-hexahydro- 2H-pyrazino[1,2-a]indolone 147 2-[3'-Hydroxymethyl 554.64 0.32 N methyl(5-methyl-4,5,6,7- tetrahydro-pyrazolo[1,5- N N a]pyrazinylamino)oxo- NH 1,6-dihydro-[3,4']bipyridinyl- N HO O 2'-yl]-2,3,5,6,7,8-hexahydro- N N 4H-2,4b-diaza-fluorenone O N 148 2-{3'-(3-Hydroxy-oxetan 692.81 5 yl)methyl[5-((S) methyloxetanylpiperazinyl )-pyridin ylamino]oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 43 149 2-{3'-Hydroxymethyl 650.77 0.0454 methyl[5-((S)methyl oxetanyl-piperazinyl)- pyridinylamino]oxo-1,6- dihydro-[3,4']bipyridinyl-2'- yl}-2,3,5,6,7,8-hexahydro-4H- 2,4b-diaza-fluorenone 150 2-[4-Hydroxymethyl-1'- 568.67 0.0316 methyl-5'-(5-methyl-4,5,6,7- tetrahydro-pyrazolo[1,5- a]pyrazinylamino)-6'-oxo- 1',6'-dihydro-[3,3']bipyridinyl- 5-yl]-7,7-dimethyl-3,4,7,8- tetrahydro-2H,6H- cyclopenta[4,5]pyrrolo[1,2- a]pyrazinone 151 2-{3'-Hydroxymethyl 648.75 0.0455 [5-((R)methyl oxetanyl-piperazinyl)- pyridinylamino]oxo-1,6- dihydro-[3,4']bipyridinyl-2'- yl}-6,7,8,9-tetrahydro-2H- pyrazino[1,2-a]indolone 152 2-{3'-Hydroxymethyl 662.78 0.188 methyl[5-((1S,5R) oxetanyl-3,8-diazabicyclo ]octyl)- nylamino]oxo-1,6- dihydro-[3,4']bipyridinyl-2'- yl}-3,4,6,7,8,9-hexahydro-2H- no[1,2-a]indolone 153 2-{3'-Hydroxymethyl 664.80 0.0238 methyl[5-((R)methyl oxetanyl-piperazinyl)- pyridinylamino]oxo-1,6- dihydro-[3,4']bipyridinyl-2'- yl}-7,7-dimethyl-3,4,7,8- tetrahydro-2H,6H- cyclopenta[4,5]pyrrolo[1,2- a]pyrazinone 44 154 3-{3'-Hydroxymethyl 666.79 0.0374 [5-((R)methyl oxetanyl-piperazinyl)- pyridinylamino]oxo-1,6- o-[3,4']bipyridinyl-2'- yl}-6,7,8,9-tetrahydro-3H- benzo[4,5]thieno[2,3- d]pyridazinone 155 2-{5-[5-((2S,5R)-2,5- 664.80 0.0454 Dimethyloxetanylpiperazinyl )-pyridin ylamino]-3'-hydroxymethyl methyloxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- 2,3,5,6,7,8-hexahydro-4H- 2,4b-diaza-fluorenone 156 2-{5-[5-((2S,5R)-2,5- 682.79 0.0145 Dimethyloxetanylpiperazinyl )-pyridin ylamino]-3'-hydroxymethyl methyloxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl} fluoro-3,4,6,7,8,9-hexahydro- 2H-pyrazino[1,2-a]indolone 157 2-{5'-[5-((2S,5R)-2,5- 664.80 0.0298 Dimethyloxetanylpiperazinyl )-pyridin ylamino]hydroxymethyl-1'- -6'-oxo-1',6'-dihydro- [3,3']bipyridinylyl}- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 158 2-{5-[5-((2S,5R)-2,5- 678.82 0.020 Dimethyloxetanylpiperazinyl )-pyridin ylamino]-3'-hydroxymethyl methyloxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}-7,7- dimethyl-3,4,7,8-tetrahydro- 2H,6H- cyclopenta[4,5]pyrrolo[1,2- a]pyrazinone 45 159 3-{5-[5-((2S,5R)-2,5- 680.82 0.082 Dimethyloxetanylpiperazinyl )-pyridin ylamino]-3'-hydroxymethyl methyloxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- 6,7,8,9-tetrahydro-3H- benzo[4,5]thieno[2,3- d]pyridazinone 160 2-{5-[5-((2S,5R)-2,5- 662.78 0.0547 Dimethyloxetanylpiperazinyl )-pyridin o]-3'-hydroxymethyl methyloxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- 6,7,8,9-tetrahydro-2H- pyrazino[1,2-a]indolone 161 O 2-{5-[5-((S)Ethyloxetan- 664.80 0.064 N iperazinyl)-pyridin ylamino]-3'-hydroxymethyl N methyloxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- N NH 2,3,5,6,7,8-hexahydro-4H- N OH O 2,4b-diaza-fluorenone N N O N 162 2-[3'-Hydroxymethyl 497.55 0.434 methyloxo(pyrazin ylamino)-1,6-dihydro- [3,4']bipyridinyl-2'-yl]- 3,4,6,7,8,9-hexahydro-2H- pyrazino[1,2-a]indolone 163 O 2-[3'-Hydroxymethyl 610.71 0.0228 methyl(5-oxetanyl- 4,5,6,7-tetrahydro- N pyrazolo[1,5-a]pyrazin ylamino)oxo-1,6-dihydro- N N [3,4']bipyridinyl-2'-yl]-7,7- NH dimethyl-3,4,7,8-tetrahydro- HO O N 2H,6H- N N enta[4,5]pyrrolo[1,2- a]pyrazinone O N 46 164 2-{5-[5-((S)Ethyloxetan- 678.82 0.029 O 3-yl-piperazinyl)-pyridin N o]-3'-hydroxymethyl N methyloxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}-7,7- N NH dimethyl-3,4,7,8-tetrahydro- HO O N 2H,6H- cyclopenta[4,5]pyrrolo[1,2- N N a]pyrazinone O N 165 2-{5-[5-((S)Ethyloxetan- 662.78 0.0417 iperazinyl)-pyridin ylamino]-3'-hydroxymethyl oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- 6,7,8,9-tetrahydro-2H- pyrazino[1,2-a]indolone 166 10-Fluoro[3'- 614.67 0.155 hydroxymethylmethyl(5- oxetanyl-4,5,6,7- tetrahydro-pyrazolo[1,5- a]pyrazinylamino)oxo- 1,6-dihydro-[3,4']bipyridinyl- 2'-yl]-3,4,6,7,8,9-hexahydro- 2H-pyrazino[1,2-a]indolone 167 hydroxymethyl)(1- 566.65 0.119 methyl(5-methyl-4,5,6,7- tetrahydropyrazolo[1,5- a]pyrazinylamino)oxo- 1,6-dihydropyridin yl)pyridinyl)-3,4,6,7,8,9- hexahydro-6,9- methanopyrazino[1,2-a]indol- 1(2H)-one 168 O 2-[5-(6,7-Dihydro-4H- 555.63 0.0635 pyrazolo[5,1-c][1,4]oxazin N ylamino)-3'-hydroxymethyl N NH methyloxo-1,6-dihydro- HO O [3,4']bipyridinyl-2'-yl]-7,7- N dimethyl-3,4,7,8-tetrahydro- N N 2H,6H- cyclopenta[4,5]pyrrolo[1,2- O N a]pyrazinone 47 169 2-{3'-Hydroxymethyl 653.79 0.206 methyl[5-(4-oxetanylpiperazinyl )-pyridin ylamino]oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- 3,4,5,6,7,8-hexahydro-2H- benzo[4,5]thieno[2,3- c]pyridinone 170 2-(3-(hydroxymethyl)(1- 566.65 0.335 N methyl(5-methyl-4,5,6,7- N tetrahydropyrazolo[1,5- N NH zinylamino)oxo- HO O 1,6-dihydropyridin N yl)pyridinyl)-3,4,6,7,8,9- N N hexahydro-6,9- methanopyrazino[1,2-a]indol- O N 1(2H)-one 171 O R)[3- 662.78 0.036 N (Hydroxymethyl)[1- N methyl({5-[(2S)methyl- 4-(oxetanyl)piperazin N NH idinyl}amino)oxo- HO O 1,6-dihydropyridin N yl]pyridinyl]-3,6- N N diazatetracyclo[9.2.1.02,10.03,8] tetradeca-2(10),8-dienone O N 172 2-(4-(5-(1,2,4-triazin 498.54 5 ylamino)methyloxo-1,6- dihydropyridinyl) (hydroxymethyl)pyridinyl)- 3,4,6,7,8,9- hexahydropyrazino[1,2- a]indol-1(2H)-one 173 2-[5-(2,6-Dimethyl-pyrimidin- 539.63 1 ino)-3'-hydroxymethyl- 1-methyloxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl]-7,7- dimethyl-3,4,7,8-tetrahydro- 2H,6H- cyclopenta[4,5]pyrrolo[1,2- a]pyrazinone 48 174 (1R,11S)[3- 662.78 0.101 (Hydroxymethyl)[1- methyl({5-[(2S)methyl- 4-(oxetanyl)piperazin yl]pyridinyl}amino)oxo- hydropyridin yl]pyridinyl]-3,6- diazatetracyclo[9.2.1.02,10.03,8] tetradeca-2(10),8-dienone 175 O 3-{5-[5-((S)Ethyloxetan- 680.82 0.0466 N 3-yl-piperazinyl)-pyridin ylamino]-3'-hydroxymethyl N methyloxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}- N NH 6,7,8,9-tetrahydro-3H- OH O N benzo[4,5]thieno[2,3- N N d]pyridazinone S O N 176 (S)(3-(hydroxymethyl) 648.75 0.0375 (1-methyl(5-(2-methyl (oxetanyl)piperazin yl)pyridinylamino)oxo- 1,6-dihydropyridin yl)pyridinyl)-6,7,8,9- tetrahydropyrazino[1,2- a]indol-1(2H)-one 177 2-(4-(5-(5-ethyl-4,5,6,7- 568.67 0.107 ydropyrazolo[1,5- a]pyrazinylamino) methyloxo-1,6- dihydropyridinyl) (hydroxymethyl)pyridinyl)- 7,8,9- hexahydropyrazino[1,2- a]indol-1(2H)-one 178 3-[3'-Hydroxymethyl 512.58 1.1 methyloxo(pyridin o)-1,6-dihydro- [3,4']bipyridinyl-2'-yl]-6,7,8,9- tetrahydro-3H- benzo[4,5]thieno[2,3- d]pyridazinone 49 179 2-[3'-Hydroxymethyl 525.60 0.209 methyl(2-methylpyrimidinylamino )oxo- 1,6-dihydro-[3,4']bipyridinyl- -7,7-dimethyl-3,4,7,8- tetrahydro-2H,6H- cyclopenta[4,5]pyrrolo[1,2- a]pyrazinone 180 2-[3'-Hydroxymethyl 525.60 0.245 methyl(6-methylpyrimidinylamino o- 1,6-dihydro-[3,4']bipyridinyl- 2'-yl]-7,7-dimethyl-3,4,7,8- tetrahydro-2H,6H- cyclopenta[4,5]pyrrolo[1,2- a]pyrazinone 181 Hydroxymethyl 570.67 0.144 (5-methyl-4,5,6,7- tetrahydro-pyrazolo[1,5- a]pyrazinylamino)oxo- 1,6-dihydro-[3,4']bipyridinyl- 2'-yl]-6,7,8,9-tetrahydro-3H- benzo[4,5]thieno[2,3- d]pyridazinone 182 3-[3'-Hydroxymethyl 513.57 0.813 methyloxo(pyrimidin ylamino)-1,6-dihydro- [3,4']bipyridinyl-2'-yl]-6,7,8,9- tetrahydro-3H- benzo[4,5]thieno[2,3- d]pyridazinone 183 10-fluoro(3- 514.55 0.906 (hydroxymethyl)(1-methyl- N NH 6-oxo(pyridinylamino)- HO N O 1,6-dihydropyridin yl)pyridinyl)-3,4,6,7,8,9- N N hexahydropyrazino[1,2- F O N a]indol-1(2H)-one 184 6-[3'-Hydroxymethyl 528.63 0.601 methyloxo(pyrazin ylamino)-1,6-dihydro- [3,4']bipyridinyl-2'-yl]-2,2- dimethyl-2,3,5,6-tetrahydro- 1H,4Hthiaazacyclopenta [a]indenone 50 185 2-{3-Hydroxymethyl[6- 550.61 1.3 (imidazo[1,2-a]pyridin ylamino)methyloxo-4,5- dihydro-pyrazinyl]-pyridin- 2-yl}-7,7-dimethyl-3,4,7,8- tetrahydro-2H,6H- cyclopenta[4,5]pyrrolo[1,2- a]pyrazinone 186 10-fluoro(3- 515.54 1.6 (hydroxymethyl)(4-methyl- 5-oxo(pyridinylamino)- 4,5-dihydropyrazin yl)pyridinyl)-3,4,6,7,8,9- hexahydropyrazino[1,2- a]indol-1(2H)-one 187 O 2-(4-(5-(5-(2,2-dimethyl 664.80 0.0451 N (oxetanyl)piperazin N yl)pyridinylamino) methyloxo-1,6- N NH dihydropyridinyl) HO O (hydroxymethyl)pyridinyl)- N 3,4,6,7,8,9- N N dropyrazino[1,2- O N a]indol-1(2H)-one 188 2-[3'-Hydroxymethyl 511.57 0.601 oxo(pyrazin ylamino)-1,6-dihydro- [3,4']bipyridinyl-2'-yl]-7,7- dimethyl-3,4,7,8-tetrahydro- 2H,6H- cyclopenta[4,5]pyrrolo[1,2- a]pyrazinone 189 2-(3-(hydroxymethyl)(1- 637.73 0.652 methyl(5-(4-(oxetan erazinyl)pyrazin o)oxo-1,6- dihydropyridinyl)pyridin yl)-3,4,6,7,8,9- hexahydropyrazino[1,2- a]indol-1(2H)-one Table 2.
No. Structure IUPAC Name CD69 Hu Blood FACS (IC70) 51 190 2-[4-[5-[[5-[2,2-dimethyl 0.0704 nyl)piperazinyl] pyridyl]amino]methyloxo- 3-pyridyl](hydroxymethyl) pyridyl]-3,4,6,7,8,9- dropyrazino[1,2-a]indol- 1-one 191 3-[3-(hydroxymethyl)[1- 0.0435 methyl[[5-[(2S)methyl (oxetanyl)piperazinyl] pyridyl]amino]oxo pyridyl]pyridyl]-6,7,8,9- tetrahydrobenzothiopheno[2,3- d]pyridazinone 192 3-[3-(hydroxymethyl)[1- 1.1 methyl[[5-[(2S)methyl (oxetanyl)piperazinyl] pyridyl]amino]oxo pyridyl]pyridyl]-7,8,9,10- tetrahydropyridazino[4,5- a]indolizinone 193 2-[3-(hydroxymethyl)[1- 0.0995 methyl[[5-[(2S)methyl (oxetanyl)piperazinyl] pyridyl]amino]oxo pyridyl]pyridyl]-6,7,8,9- tetrahydropyridazino[4,5- b]indolizinone 194 3-[3-(hydroxymethyl)[1- 1.2 methyloxo(pyrazin ylamino)pyridyl]pyridyl]- 6,7,8,9- tetrahydrobenzothiopheno[2,3- dazinone 195 2-[3-(hydroxymethyl)[1- 0.101 methyl[(5-methyl-6,7- dihydro-4H-pyrazolo[1,5- a]pyrazinyl)amino]oxo pyridyl]pyridyl]-6,7,8,9- tetrahydropyrazino[1,2-a]indol- 1-one 52 196 3-[4-[5-[(2- 0.325 cyclopropylpyrimidin yl)amino]methyloxo pyridyl](hydroxymethyl) pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 197 O 2-[3-(hydroxymethyl)[4- 2.3 N [[6-[4-(oxetan N yl)piperazinyl] pyridyl]amino]oxo-pyrazin N yl]pyridyl]-3,4,6,7,8,9- NH hexahydropyrazino[1,2-a]indol- OH O N N 1-one N N O N 198 3-[3-(hydroxymethyl)[1- 6 methyl[[5-[4-(oxetan yl)piperazinyl] pyridyl]amino]oxo pyridyl]pyridyl]-7,8,9,10- tetrahydropyridazino[4,5- a]indolizinone 199 N 3-[3-(hydroxymethyl)[5- 0.934 (imidazo[1,2-a]pyrimidin N N NH o)methyloxo HO O pyridyl]pyridyl]-7,7- N yl-1,2,6,8- N N tetrahydrocyclopenta[3,4]pyrrolo O N [3,5-b]pyrazinone 200 2-[3-(hydroxymethyl)[1- 0.636 methyl[[5-[1-(oxetanyl)- 3,6-dihydro-2H-pyridinyl] pyridyl]amino]oxo pyridyl]pyridyl]-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol- 1-one 53 201 3-[3-(hydroxymethyl)[6- 3.3 (imidazo[1,2-a]pyridin ylamino)methyloxopyrazinyl ]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo ]pyrazinone 202 hydroxymethyl)[1- 7.3 N methyl[(5-methyl-6,7- dihydro-4H-pyrazolo[1,5- N N NH a]pyrazinyl)amino]oxo HO O pyridyl]pyridyl]-7,8,9,10- N tetrahydropyridazino[4,5- N N N a]indolizinone O N 203 3-[3-(hydroxymethyl)[1- 0.0605 methyl[(5-methyl-6,7- o-4H-thiazolo[5,4- c]pyridinyl)amino]oxo pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 204 2-[3-(hydroxymethyl)[1- 0.436 [(5-methyl-6,7- dihydro-4H-pyrazolo[1,5- a]pyrazinyl)amino]oxo pyridyl]pyridyl]-6,7,8,9- tetrahydropyridazino[4,5- b]indolizinone 205 2-[3-(hydroxymethyl)[1- 0.114 methyloxo(4,5,6,7- tetrahydropyrazolo[1,5- a]pyrazinylamino)pyridyl]- 2-pyridyl]-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol- 1-one 206 2-[3-(hydroxymethyl)[1- 0.15 methyl[[5-(1-methylazetidin- 3-yl)oxypyridyl]amino] oxopyridyl]pyridyl]- 3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol- 1-one 54 207 O 3-[3-(hydroxymethyl)[1- 0.0414 N methyl[[5-(1-methylazetidin- N NH 3-yl)oxypyridyl]amino] OH O oxopyridyl]pyridyl]-7,7- N dimethyl-1,2,6,8- N N tetrahydrocyclopenta[3,4]pyrrolo O N [3,5-b]pyrazinone 208 2-[4-[5-[(5-ethyl-6,7-dihydro- 0.58 N 4H-pyrazolo[1,5-a]pyrazin yl)amino]methyloxo N pyridyl](hydroxymethyl) N NH pyridyl]-3,4,6,7,8,9- N OH O hexahydropyrido[3,4- N N b]indolizinone O N 209 2-[4-[5-[(5-acetyl-6,7-dihydro- 0.116 4H-pyrazolo[1,5-a]pyrazin yl)amino]methyloxo pyridyl](hydroxymethyl) pyridyl]-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol- 1-one 210 2-(7,7-dimethyloxo-1,2,6,8- 0.914 tetrahydrocyclopenta[3,4]pyrrolo ]pyrazinyl)[1- methyl[[5-[(2S)methyl (oxetanyl)piperazinyl] pyridyl]amino]oxo pyridyl]pyridinecarboxamide 211 2-(7,7-dimethyloxo-1,2,6,8- 2.1 tetrahydrocyclopenta[3,4]pyrrolo ]pyrazinyl)-N-methyl- 4-[1-methyl[[5-[(2S) methyl(oxetanyl)piperazin- 2-pyridyl]amino]oxo pyridyl]pyridinecarboxamide 212 3-[4-[5-[(5-acetyl-6,7-dihydro- 0.0152 azolo[1,5-a]pyrazin yl)amino]methyloxo pyridyl](hydroxymethyl) pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 55 213 10-fluoro[3-(hydroxymethyl)- 0.62 4-[1-methyl[(2- methylpyrimidinyl)amino] oxopyridyl]pyridyl]- 7,8,9- hexahydropyrazino[1,2-a]indol- 1-one 214 3-[4-[5-(6,7-dihydro-4H- 0.124 pyrano[4,3-d]thiazol ylamino)methyloxo pyridyl](hydroxymethyl) pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 215 3-[4-(hydroxymethyl)[1- 0.457 methyl[(2-methylpyrimidin- 4-yl)amino]oxopyridyl] pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 216 3-[3-(hydroxymethyl)[1- 0.357 N methyl[(5-methyl-6,7- S dihydro-4H-thiazolo[5,4- c]pyridinyl)amino]oxo N NH pyridyl]pyridyl]-6,7,8,9- HO O N tetrahydrobenzothiopheno[2,3- N N dazinone S O N 217 2-[3-(hydroxymethyl)[5-(1H- 2.9 imidazo[4,5-b]pyridin ylamino)methyloxo pyridyl]pyridyl]-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol- 1-one 218 3-[4-[5-[(1,5-dimethylpyrazol 0.0741 yl)amino]methyloxo l](hydroxymethyl) pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 56 219 3-[4-[6-(3-aminoanilino) 0.204 methyloxo-pyrazinyl] (hydroxymethyl)pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 220 5-[2-(3,4,6,7,8,9-hexahydro-1H- 1.6 pyrazino[1,2-a]indolyl) (hydroxymethyl)pyridyl] methyl[[5-[(2S)methyl (oxetanyl)piperazinyl] l]amino]pyridinone 221 3-[4-[5-[(6-amino 0.121 pyridyl)amino]methyloxo- 3-pyridyl](hydroxymethyl) l]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 222 3-[3-(hydroxymethyl)[5-[(2- 0.178 methylpyrimidinyl)amino] oxo-1H-pyridinyl]pyridyl]- 7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 223 10-fluoro[3-(hydroxymethyl)- 0.43 N 4-[1-methyl[(5-methyl-6,7- S dihydro-4H-thiazolo[5,4- c]pyridinyl)amino]oxo N NH pyridyl]pyridyl]-3,4,6,7,8,9- HO O hexahydropyrazino[1,2-a]indol- N 1-one N N F O N 224 3-[3-(hydroxymethyl)[1- 0.0307 methyl[[5-[(2S)methyl (oxetanyl)piperazinyl] pyridyl]amino]oxo-pyridazin- 3-yl]pyridyl]-7,7-dimethyl- 1,2,6,8- ydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 57 225 3-[3-(hydroxymethyl)[1- 0.766 methyl[(2-methylpyrimidin- 4-yl)amino]oxo-pyridazin yl]pyridyl]-7,7-dimethyl- 1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 226 3-[3-(hydroxymethyl)[1- 0.117 methyl[[5-(morpholine carbonyl)pyridyl]amino] oxo-pyridazinyl]pyridyl]- 7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 227 O hydroxymethyl)[1- 0.73 methyl[[5-(morpholine N carbonyl)pyridyl]amino] O N NH oxo-pyridazinyl]pyridyl]- OH O 3,4,6,7,8,9- N hexahydropyrazino[1,2-a]indol- N N N 1-one O N 228 2-[3-(hydroxymethyl)[1- 0.369 methyl[[5-[(2S)methyl nyl)piperazin yl]pyrazinyl]amino]oxo pyridyl]pyridyl]-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol- 1-one 229 3-[4-[5-[(2-ethylpyrimidin 0.583 yl)amino]methyloxo pyridyl](hydroxymethyl) pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 230 O 3-[3-(hydroxymethyl)[1- 0.179 N [[5-[(2S)methyl N N (oxetanyl)piperazin yl]pyrazinyl]amino]oxo pyridyl]pyridyl]-6,7,8,9- N NH tetrahydrobenzothiopheno[2,3- OH O N d]pyridazinone S N N O N 58 231 O 10-fluoro[3-(hydroxymethyl)- 0.0624 N 4-[1-methyl[[5-[(2S) N methyl(oxetanyl)piperazin- 1-yl]pyridyl]amino]oxo- N NH pyridazinyl]pyridyl]- OH O 3,4,6,7,8,9- N hexahydropyrazino[1,2-a]indol- N N N 1-one F O N 232 2-[3-(hydroxymethyl)[1- 0.0518 methyl[[5-(1-methyl dyl)pyridyl]amino] oxopyridyl]pyridyl]- 3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol- 1-one 233 hydroxymethyl)[1- 0.0657 methyloxo(4,5,6,7- tetrahydrothiazolo[5,4-c]pyridin- 2-ylamino)pyridyl] pyridyl]-7,7-dimethyl-1,2,6,8- ydrocyclopenta[3,4]pyrrolo ]pyrazinone 234 2-[4-[5-[(5-acetyl-6,7-dihydro- 0.183 4H-thiazolo[5,4-c]pyridin yl)amino]methyloxo pyridyl](hydroxymethyl) pyridyl]fluoro-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol- 1-one 235 3-[3-(hydroxymethyl)[1- 0.112 methyl[[5-[(2S)methyl (oxetanyl)piperazin yl]pyrazinyl]amino]oxo pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 236 N 3-[3-(hydroxymethyl)[1- 0.0336 methyl[[5-(1-methyl piperidyl)pyridyl]amino] N NH oxopyridyl]pyridyl]-7,7- OH O dimethyl-1,2,6,8- N tetrahydrocyclopenta[3,4]pyrrolo N N [3,5-b]pyrazinone O N 59 237 10-fluoro[3-(hydroxymethyl)- 0.0461 4-[1-methyl[[5-[(2S) methyl(oxetanyl)piperazin- 1-yl]pyridyl]amino]oxo pyridyl]pyridyl]-3,4,6,7,8,9- hexahydropyrido[3,4- b]indolizinone 238 2-[3-(hydroxymethyl)[1- 5 methyl[[5-[(2S)methyl (oxetanyl)piperazinyl] pyridyl]amino]oxo pyridyl]pyridyl]-5,6,7,8- tetrahydro-1H-pyrrolo[3,4- b]indolizinone 239 2-[3-(hydroxymethyl)[1- 0.153 methyl[[5-(1-methylazetidin- 3-yl)pyridyl]amino]oxo pyridyl]pyridyl]-3,4,6,7,8,9- dropyrazino[1,2-a]indol- 1-one 240 3-[3-(hydroxymethyl)[1- 0.0229 methyl[[5-(1-methylazetidin- 3-yl)pyridyl]amino]oxo pyridyl]pyridyl]-7,7- yl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 241 10-fluoro[3-(hydroxymethyl)- 0.19 4-[1-methyl[(5-methyl-6,7- dihydro-4H-pyrazolo[1,5- a]pyrazinyl)amino]oxo pyridyl]pyridyl]-3,4,6,7,8,9- hexahydropyrido[3,4- lizinone 242 2-[4-[5-[(1,5-dimethylpyrazol 1.2 yl)amino]methyloxo pyridyl](hydroxymethyl) pyridyl]-3,4,6,7,8,9- hexahydropyrido[3,4- b]indolizinone 60 243 2-[3-(hydroxymethyl)[1- 2.8 methyl[[1-[1-(oxetanyl) dyl]imidazolyl]amino]- 6-oxopyridyl]pyridyl]- 3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol- 1-one 244 2-[3-(hydroxymethyl)[1- 0.138 methyl[[5-(1-methylazetidin- 3-yl)pyridyl]amino]oxo l]pyridyl]-3,4,6,7,8,9- hexahydropyrido[3,4- b]indolizinone 245 2-[3-(hydroxymethyl)[1- 0.065 methyl[[5-[(2S)methyl (oxetanyl)piperazinyl] pyridyl]amino]oxo pyridyl]pyridyl]-6,7,8,9- tetrahydropyrido[3,4- b]indolizinone 246 2-[4-[5-[(1,5-dimethylpyrazol 1.7 yl)amino]methyloxo l](hydroxymethyl) pyridyl]-6,7,8,9- tetrahydropyrido[3,4- b]indolizinone 247 3-[4-[5-[(5-acetyl-6,7-dihydro- 0.145 4H-pyrazolo[1,5-a]pyrazin yl)amino]methyloxo pyridyl](hydroxymethyl) pyridyl]-6,7,8,9- tetrahydrobenzothiopheno[2,3- d]pyridazinone 61 248 3-[3-(hydroxymethyl)[1- 0.0703 methyl[[5-[(3R) methylmorpholinecarbonyl]- 2-pyridyl]amino]oxopyridazinyl ]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 249 3-[3-(hydroxymethyl)[1- 0.0177 methyl[[5-[(3R) methylmorpholinecarbonyl]- 2-pyridyl]amino]oxo pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- ydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 250 2-[4-[5-[(5,6-dimethyl-6,7- 0.171 dihydro-4H-pyrazolo[1,5- a]pyrazinyl)amino]methyl- 6-oxopyridyl] (hydroxymethyl)pyridyl]- 3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol- 1-one 251 252 3-[4-[5-[(1-ethylmethyl- 0.252 pyrazolyl)amino]methyl oxopyridyl] (hydroxymethyl)pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 253 hydroxymethyl)[5-[[5- 0.0164 2-methyl(oxetan yl)piperazinyl] pyridyl]amino]oxo-1H- pyridazinyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 62 254 3-[4-[5-[(5-acetyl-6,7-dihydro- 0.0373 4H-pyrazolo[1,5-a]pyrazin yl)amino]methyloxo pyridyl](hydroxymethyl) pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]thieno[ 1,3-c]pyridinone 255 O 2-[4-[5-[(5-acetyl-6,7-dihydro- 0.094 4H-pyrazolo[1,5-a]pyrazin N no]methyloxo pyridyl](hydroxymethyl) N pyridyl]fluoro-3,4,6,7,8,9- N NH hexahydropyrazino[1,2-a]indol- HO O N 1-one N N F O N 256 3-[3-(hydroxymethyl)[1- 0.08 methyl[[5-[(3S) methylmorpholinecarbonyl]- 2-pyridyl]amino]oxopyridazinyl ]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 257 3-[3-(hydroxymethyl)[1- 0.0216 methyl[[5-[(3S) methylmorpholinecarbonyl]- 2-pyridyl]amino]oxo pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 258 3-[3-(hydroxymethyl)[1- 0.646 [[5-(morpholine carbonyl)pyridyl]amino] oxo-pyridazinyl]pyridyl]- 6,7,8,9- tetrahydrobenzothiopheno[2,3- dazinone 259 hydroxymethyl)[1- 0.301 methyl[[5-(morpholine carbonyl)pyridyl]amino] oxo-pyridazinyl]pyridyl]- 7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]thieno[ 1,3-c]pyridinone 63 260 3-[3-(hydroxymethyl)[1- 0.0606 methyl[(5-methyl-1H- pyrazolyl)amino]oxo pyridyl]pyridyl]-7,7- yl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 261 2-[4-[5-[(1,5-dimethylpyrazol 2.9 yl)amino]methyloxo N N pyridyl](hydroxymethyl) NH pyridyl]fluoro-3,4,6,7,8,9- N HO O dropyrido[3,4- N N b]indolizinone F O N 262 3-[3-(hydroxymethyl)[1- 0.577 methyl[(5-methyloxazol yl)amino]oxopyridyl] pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 263 N 10-fluoro[3-(hydroxymethyl)- 2.2 4-[1-methyloxo(pyrimidin- N NH 4-ylamino)pyridazinyl] HO N O pyridyl]-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol- N N N 1-one F O N 264 N 3-[3-(hydroxymethyl)[5-[[2- 0.653 HO roxymethyl- N NH ethyl)pyrimidinyl]amino] OH O methyloxopyridyl] N pyridyl]-7,7-dimethyl-1,2,6,8- N N tetrahydrocyclopenta[3,4]pyrrolo O N [3,5-b]pyrazinone 265 3-[3-(hydroxymethyl)[1- 0.0091 methyloxo[(5-propanoyl- 6,7-dihydro-4H-pyrazolo[1,5- a]pyrazinyl)amino] pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 64 266 3-[3-(hydroxymethyl)[1- 0.0293 methyl[[5-(oxetanyl)-1H- pyrazolyl]amino]oxo pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 267 3-[3-(hydroxymethyl)[1- 0.225 methyl[(5-methyl-1,3,4- thiadiazolyl)amino]oxo pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 268 hydroxymethyl)[1- 0.212 methyl[(1-methylimidazol yl)amino]oxopyridyl] pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo ]pyrazinone 269 O 3-[3-(hydroxymethyl)[1- 0.0251 N methyl[[5-(morpholine O carbonyl)pyridyl]amino] N NH oxopyridyl]pyridyl]-7,7- HO O N dimethyl-1,2,6,8- N N tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone O N 270 2-[3-(hydroxymethyl)[1- 5.9 methyl[(7-methyl-6,8- dihydro-5H-[1,2,4]triazolo[1,5- a]pyrazinyl)amino]oxo pyridyl]pyridyl]-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol- 1-one 271 O 3-[3-(hydroxymethyl)[5-[[5- 0.0245 N [(2S)methyl(oxetan N erazinyl] pyridyl]amino]oxo-1H- N NH pyridinyl]pyridyl]-7,7- HO dimethyl-1,2,6,8- O N tetrahydrocyclopenta[3,4]pyrrolo N NH [3,5-b]pyrazinone O N 65 272 3-[3-(hydroxymethyl)[1- 0.034 methyl[[5-[1-(oxetan yl)azetidinyl] l]amino]oxo pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 273 3-[3-(hydroxymethyl)[1- 0.299 methyloxo(2- pyridylamino)pyridyl] pyridyl]-7,7-dimethyl-1,2,6,8- ydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 274 hydroxymethyl)[1- 0.466 methyl[(5-methylpyrazin yl)amino]oxopyridyl] pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 275 3-[4-[5-[(5-fluoro 0.423 pyridyl)amino]methyloxo- 3-pyridyl](hydroxymethyl) pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 276 N 6-[[5-[2-(7,7-dimethyloxo- 0.358 1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo N NH [3,5-b]pyrazinyl) HO O N (hydroxymethyl)pyridyl] N N methyloxo pyridyl]amino]pyridine O N carbonitrile 277 O 3-[3-(hydroxymethyl)[5-[(5- 0.339 methoxypyridyl)amino] N NH oxopyridyl] HO O pyridyl]-7,7-dimethyl-1,2,6,8- N tetrahydrocyclopenta[3,4]pyrrolo N N [3,5-b]pyrazinone O N 66 278 3-[4-[5-[(5-cyclopropyl 3.2 pyridyl)amino]methyloxo- 3-pyridyl](hydroxymethyl) pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 279 3-[3-(hydroxymethyl)[1- 2.1 methyloxo[[5- (trifluoromethyl) pyridyl]amino]pyridyl] pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo ]pyrazinone 280 3-[3-(hydroxymethyl)[1- 0.0141 [[1-methyl (morpholinecarbonyl)pyrazol- mino]oxopyridyl] pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 281 3-[3-(hydroxymethyl)[1- 0.718 methyl[(5-methyl pyridyl)amino]oxo l]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 282 3-[3-(hydroxymethyl)[1- 0.0174 methyl[[5-[(2S)methyl (oxetanyl)piperazinyl] pyridyl]amino]oxo pyridyl]pyridyl]-7,7- dimethyl-6,8- dihydrocyclopenta[3,4]thieno[1, 3-d]pyridazinone 283 3-[3-(hydroxymethyl)[1- 0.143 methyl[(5-methylisoxazol yl)amino]oxopyridyl] pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 67 284 10-fluoro[3-(hydroxymethyl)- 0.131 4-[1-methyl[[5-(oxetanyl)- 6,7-dihydro-4H-pyrazolo[1,5- a]pyrazinyl]amino]oxo pyridyl]pyridyl]-3,4,6,7,8,9- hexahydropyrido[3,4- b]indolizinone 285 3-[3-(hydroxymethyl)[1- 0.175 [[1-(oxetan yl)imidazolyl]amino]oxo- 3-pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 286 hydroxymethyl)[5- 0.167 (isoxazolylamino)methyl- 6-oxopyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 287 2-[4-[5-[(4,5-dimethyl-6,7- 0.127 dihydro-4H-pyrazolo[1,5- a]pyrazinyl)amino]methyl- 6-oxopyridyl] (hydroxymethyl)pyridyl]- 3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol- 1-one 288 3-[3-(hydroxymethyl)[1- 0.229 [(5-methylisoxazol yl)amino]oxopyridyl] pyridyl]-7,7-dimethyl-6,8- dihydrocyclopenta[3,4]thieno[1, 3-d]pyridazinone 289 3-[3-(hydroxymethyl)[1- 0.214 methyloxo(1H-pyrazol ylamino)pyridyl]pyridyl]- 7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 68 290 3-[3-(hydroxymethyl)[1- 0.113 methyl[(1-methylpyrazol yl)amino]oxopyridyl] pyridyl]-7,7-dimethyl-1,2,6,8- ydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 291 3-[3-(hydroxymethyl)[1- 0.843 methyl[(5-methylisoxazol yl)amino]oxopyridyl] pyridyl]-6,7,8,9- tetrahydrobenzothiopheno[2,3- d]pyridazinone 292 3-[4-[5-[(1,5-dimethylpyrazol 0.118 yl)amino]methyloxo pyridyl](hydroxymethyl) pyridyl]-7,7-dimethyl-6,8- dihydrocyclopenta[3,4]thieno[1, 3-d]pyridazinone 293 3-[3-(hydroxymethyl)[1- 0.0691 methyl[(1-methyltriazol yl)amino]oxopyridyl] pyridyl]-7,7-dimethyl-1,2,6,8- ydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 294 3-[4-[5-[(5-tert-butylisoxazol 0.276 yl)amino]methyloxo pyridyl](hydroxymethyl) pyridyl]-7,7-dimethyl-1,2,6,8- ydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 295 3-[4-[5-[(5-ethylisoxazol 0.134 yl)amino]methyloxo pyridyl](hydroxymethyl) pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 69 296 3-[3-(hydroxymethyl)[5-[[5- 0.0193 hoxyethyl)-6,7-dihydro- 4H-pyrazolo[1,5-a]pyrazin yl]amino]methyloxo pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 297 3-[3-(hydroxymethyl)[1- 0.14 methyl[(5-methyl-6,7- dihydro-4H-pyrazolo[1,5- a]pyrazinyl)amino]oxopyridazinyl ]pyridyl]-7,7- dimethyl-1,2,6,8- ydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 298 5-[[5-[2-(7,7-dimethyloxo- 0.869 1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinyl) (hydroxymethyl)pyridyl] methyloxo pyridyl]amino]pyrazine carbonitrile 299 hydroxymethyl)[1- 2.1 methyloxo[(5- phenylisoxazolyl)amino] l]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 300 (R)(3'-(hydroxymethyl)((5- 0.024 (1-methoxypropanyl)-4,5,6,7- tetrahydropyrazolo[1,5- a]pyrazinyl)amino)methyl- 6-oxo-1,6-dihydro-[3,4'- bipyridin]-2'-yl)-7,7-dimethyl- 2,3,4,6,7,8-hexahydro-1H- cyclopenta[4,5]pyrrolo[1,2- a]pyrazinone 70 301 3-[3-(hydroxymethyl)[1- 1.3 methyloxo[[6- (trifluoromethyl)pyridazin no]pyridyl]pyridyl]- 7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 302 O 3-[3-(hydroxymethyl)[1- 0.0228 methyl[[1-methyl N [[methyl(oxetan yl)amino]methyl]pyrazol yl]amino]oxopyridyl] N HN N pyridyl]-7,7-dimethyl-1,2,6,8- HO O tetrahydrocyclopenta[3,4]pyrrolo N [3,5-b]pyrazinone N N O N 303 (S)(3'-(hydroxymethyl)((5- 0.0179 (1-methoxypropanyl)-4,5,6,7- tetrahydropyrazolo[1,5- a]pyrazinyl)amino)methyl- 6-oxo-1,6-dihydro-[3,4'- bipyridin]-2'-yl)-7,7-dimethyl- 2,3,4,6,7,8-hexahydro-1H- cyclopenta[4,5]pyrrolo[1,2- a]pyrazinone 304 3-[3-(hydroxymethyl)[5-[[5- 0.04 (2-methoxyethyl)-6,7-dihydro- 4H-pyrazolo[1,5-a]pyrazin yl]amino]methyloxo pyridyl]pyridyl]-7,7- yl-6,8- dihydrocyclopenta[3,4]thieno[1, 3-d]pyridazinone 305 3-[3-(hydroxymethyl)[5-[[5- 0.0832 (2-methoxyethyl)-6,7-dihydro- azolo[1,5-a]pyrazin yl]amino]methyloxopyridazinyl ]pyridyl]-7,7- yl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 71 306 3-[3-(hydroxymethyl)[5-[(6- 0.602 methoxypyridazinyl)amino]- 1-methyloxopyridyl] pyridyl]-7,7-dimethyl-1,2,6,8- ydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 307 N 3-[4-[6-[(1,3-dimethylindazol N yl)amino]methyloxopyrazinyl ] (hydroxymethyl)pyridyl]-7,7- NH dimethyl-1,2,6,8- HO O N N tetrahydrocyclopenta[3,4]pyrrolo N N [3,5-b]pyrazinone O N 308 3-[3-(hydroxymethyl)[5-[[5- 0.0546 (2-methoxyethyl)-6,7-dihydro- 4H-pyrazolo[1,5-a]pyrazin no]methyloxo pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]thieno[ 1,3-c]pyridinone 309 3-[3-(hydroxymethyl)[5-[[5- 0.0398 (3-methoxypropyl)-6,7-dihydro- 4H-pyrazolo[1,5-a]pyrazin yl]amino]methyloxo pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 310 3-[3-(hydroxymethyl)[1- 0.119 methyl[(5-methylisothiazol S yl)amino]oxopyridyl] N NH l]-7,7-dimethyl-1,2,6,8- HO O N tetrahydrocyclopenta[3,4]pyrrolo N N [3,5-b]pyrazinone O N 72 311 3-[4-[5-[(5-cyclopropylisoxazol- 0.158 3-yl)amino]methyloxo pyridyl](hydroxymethyl) pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 312 hydroxymethyl)[1- 5.6 methyl[[5-methyl(oxetan- 3-yl)pyrazolyl]amino]oxo- 3-pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 313 3-[3-(hydroxymethyl)[1- 0.0191 methyl[(5-methyloxo-4,7- dihydropyrazolo[1,5-a]pyrazin- 2-yl)amino]oxopyridyl] pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 314 3-[4-[5-[[5-(3-hydroxyazetidin- 0.0446 1-yl)pyridyl]amino] methyloxopyridyl] (hydroxymethyl)pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 315 O hydroxymethyl)[1- 0.015 N [[1-methyl (pyrrolidinecarbonyl)pyrazol- N 3-yl]amino]oxopyridyl] HN N pyridyl]-7,7-dimethyl-1,2,6,8- HO O N tetrahydrocyclopenta[3,4]pyrrolo N N [3,5-b]pyrazinone O N 316 hydroxymethyl)[5-[[5- 0.0202 (methoxymethyl)methylpyrazolyl ]amino]methyl oxopyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 73 317 3-[3-(hydroxymethyl)[1- 0.0586 [(5-methyloxo-4,7- dihydropyrazolo[1,5-a]pyrazin- 2-yl)amino]oxopyridyl] pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]thieno[ 1,3-c]pyridinone 318 (R)(5-((4,5-dimethyl-4,5,6,7- 0.108 N tetrahydropyrazolo[1,5- a]pyrazinyl)amino)-3'- N N NH (hydroxymethyl)methyl oxo-1,6-dihydro-[3,4'-bipyridin]- OH O N 2'-yl)-7,7-dimethyl-2,3,4,6,7,8- N N hexahydro-1H- cyclopenta[4,5]pyrrolo[1,2- O N a]pyrazinone 319 (S)(5-((4,5-dimethyl-4,5,6,7- 0.0167 tetrahydropyrazolo[1,5- zinyl)amino)-3'- (hydroxymethyl)methyl 6-dihydro-[3,4'-bipyridin]- 2'-yl)-7,7-dimethyl-2,3,4,6,7,8- hexahydro-1H- cyclopenta[4,5]pyrrolo[1,2- a]pyrazinone 320 3-[4-[5-[[5-[(3S,5R)-3,5- 0.0468 dimethylmorpholine carbonyl]pyridyl]amino] methyloxopyridyl] (hydroxymethyl)pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 321 O 3-[4-[5-[[5-[(3S,5R)-3,5- 0.112 dimethylmorpholine N carbonyl]pyridyl]amino] O N NH methyloxo-pyridazinyl] O (hydroxymethyl)pyridyl]-7,7- HO N dimethyl-1,2,6,8- N N tetrahydrocyclopenta[3,4]pyrrolo N ]pyrazinone O N 322 2-[4-[5-[[5-[(3S,5R)-3,5- 0.0796 dimethylmorpholine carbonyl]pyridyl]amino] methyloxopyridyl] (hydroxymethyl)pyridyl]- 3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol- 1-one 74 323 3-[3-(hydroxymethyl)[5-[[5- 0.279 (3-methoxyazetidinyl) pyridyl]amino]methyloxo- 3-pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 324 3-[4-[5-[[5-[(3S,5S)-3,5- 0.0858 dimethylmorpholine carbonyl]pyridyl]amino] methyloxo-pyridazinyl] (hydroxymethyl)pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 325 N 3-[4-[5-[(1,3- 1.4 N dimethylpyrazolo[3,4-c]pyridin- 5-yl)amino]methyloxo pyridyl](hydroxymethyl) N NH pyridyl]-7,7-dimethyl-1,2,6,8- HO O N tetrahydrocyclopenta[3,4]pyrrolo N N [3,5-b]pyrazinone O N 326 3-[4-[5-[(2,3- 1.4 ylpyrazolo[3,4-c]pyridin- 5-yl)amino]methyloxo pyridyl](hydroxymethyl) pyridyl]-7,7-dimethyl-1,2,6,8- ydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 327 3-[3-(hydroxymethyl)[5-[[5- 2.2 (2-methoxyethyl)-6,7-dihydro- 4H-pyrazolo[1,5-a]pyrazin yl]amino]-1,2-dimethyloxo pyridyl]pyridyl]-7,7- yl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 75 328 3-[4-(hydroxymethyl)[5-[[5- 0.0124 hoxyethyl)-6,7-dihydro- 4H-pyrazolo[1,5-a]pyrazin yl]amino]methyloxo pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 329 3-[3-(hydroxymethyl)[5-[[5- 0.11 (2-methoxyethyl)-6,7-dihydro- azolo[1,5-a]pyrazin yl]amino]methyloxo pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 330 3-[4-[5-[(6,6-dimethyl-4,7- 0.235 dihydropyrazolo[5,1- c][1,4]oxazinyl)amino] methyloxo-pyridazinyl] (hydroxymethyl)pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]thieno[ 1,3-c]pyridinone 331 3-[3-(hydroxymethyl)[4- methyl[(3-methylisothiazol yl)amino]oxo-pyrazinyl]- 2-pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 332 5-[(5-ethyl-1,3,4- 0.0919 thiadiazolyl)amino] methyloxopyridyl] (hydroxymethyl)pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 333 3-[3-(hydroxymethyl)[1- 0.209 methyl[(1-methyltriazol yl)amino]oxopyridyl] pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]thieno[ 1,3-c]pyridinone 76 334 3-[4-[5-[(5-cyclopropyl-1,3,4- 0.193 thiadiazolyl)amino] methyloxopyridyl] (hydroxymethyl)pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 335 5-[(6,6-dimethyl-4,7- 0.0528 dihydropyrazolo[5,1- c][1,4]oxazinyl)amino] methyloxopyridyl] (hydroxymethyl)pyridyl]-7,7- dimethyl-6,8- dihydrocyclopenta[3,4]thieno[1, 3-d]pyridazinone 336 2-[3-(hydroxymethyl)[5-[[5- 0.33 (methoxymethyl)methylpyrazolyl ]amino]methyl oxopyridyl]pyridyl]- 3,4,6,7,8,9- hexahydropyrido[3,4- lizinone 337 5-[(1,2-dimethylimidazol- 0.613 4-yl)amino]methyloxo pyridyl](hydroxymethyl) pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 338 N 3-[2-[[5-[2-(7,7-dimethyloxo- 0.0178 1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinyl) N (hydroxymethyl)pyridyl] N methyloxopyridyl]amino]- N NH 6,7-dihydro-4H-pyrazolo[1,5- HO O a]pyrazinyl]propanenitrile N N N O N 77 339 hydroxymethyl)[5-[[5- 0.0535 [4-(2-methoxyethyl)piperazin yl]pyridyl]amino]methyl- 6-oxopyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo [3,5-b]pyrazinone 340 3-[3-(hydroxymethyl)[5-[[5- 0.0207 O [(2S)(2-methoxyethyl) methyl-piperazinyl] N pyridyl]amino]methyloxo- N 3-pyridyl]pyridyl]-7,7- dimethyl-1,2,6,8- N NH tetrahydrocyclopenta[3,4]pyrrolo HO O [3,5-b]pyrazinone N N N O N ADMINISTRATION OF FORMULA I NDS The compounds of the invention may be administered by any route appropriate to the condition to be treated. Suitable routes e oral, parenteral (including subcutaneous, 5 intramuscular, intravenous, intraarterial, intradermal, intrathecal and epidural), transdermal, rectal, nasal, topical (including buccal and sublingual), vaginal, intraperitoneal, intrapulmonary and intranasal. For local immunosuppressive treatment, the compounds may be administered by intralesional stration, including perfusing or otherwise contacting the graft with the inhibitor before lantation. It will be appreciated that the preferred 10 route may vary with for example the condition of the recipient. Where the compound is stered orally, it may be formulated as a pill, capsule, tablet, etc. with a pharmaceutically acceptable carrier or excipient. Where the compound is administered parenterally, it may be formulated with a pharmaceutically acceptable parenteral vehicle and in a unit dosage injectable form, as detailed below. 15 A dose to treat human patients may range from about 10 mg to about 1000 mg of Formula I compound. A typical dose may be about 100 mg to about 300 mg of the compound. A dose may be administered once a day (QID), twice per day (BID), or more frequently, depending on the pharmacokinetic and pharmacodynamic properties, ing tion, distribution, metabolism, and excretion of the particular nd. In addition, 20 toxicity factors may influence the dosage and administration regimen. When administered 78 orally, the pill, capsule, or tablet may be ingested daily or less frequently for a specified period of time. The regimen may be repeated for a number of cycles of therapy.
METHODS OF ENT WITH FORMULA I COMPOUNDS a I compounds of the present invention are useful for ng a human or 5 animal patient suffering from a disease or disorder arising from abnormal cell growth, on or behavior associated with Btk kinase such as an immune disorder, cardiovascular disease, viral infection, inflammation, a metabolism/endocrine disorder or a neurological er, may thus be treated by a method comprising the administration thereto of a compound of the present invention as defined above. A human or animal patient suffering 10 from cancer may also be treated by a method comprising the administration thereto of a compound of the present invention as defined above. The condition of the patient may thereby be ed or ameliorated.
Formula I compounds may be useful for in vitro, in situ, and in vivo diagnosis or treatment of mammalian cells, organisms, or associated pathological conditions, such as 15 ic and local inflammation, immune-inflammatory diseases such as rheumatoid arthritis, immune suppression, organ transplant rejection, allergies, ulcerative colitis, Crohn’s disease, dermatitis, asthma, systemic lupus erythematosus, Sjögren’s Syndrome, multiple sclerosis, derma/systemic sclerosis, idiopathic ocytopenic purpura (ITP), anti-neutrophil cytoplasmic antibodies (ANCA) vasculitis, chronic obstructive pulmonary disease (COPD), 20 sis, and for general joint protective effects. s of using compounds of the ion also include treating such diseases as arthritic diseases, such as toid arthritis, monoarticular arthritis, osteoarthritis, gouty arthritis, spondylitis; Behcet disease; sepsis, septic shock, endotoxic shock, gram negative sepsis, gram positive sepsis, and toxic shock syndrome; multiple organ injury syndrome 25 secondary to septicemia, trauma, or hemorrhage; ophthalmic disorders such as allergic conjunctivitis, vernal conjunctivitis, uveitis, and thyroid-associated lmopathy; philic granuloma; pulmonary or respiratory disorders such as asthma, c itis, allergic is, ARDS, chronic pulmonary inflammatory disease (e.g., chronic obstructive pulmonary disease), silicosis, pulmonary sarcoidosis, pleurisy, alveolitis, 30 itis, emphysema, pneumonia, bronchiectasis, and pulmonary oxygen toxicity; reperfusion injury of the myocardium, brain, or extremities; fibrosis such as cystic fibrosis; keloid formation or scar tissue formation; atherosclerosis; autoimmune diseases, such as systemic lupus erythematosus (SLE), autoimmune thyroiditis, multiple sclerosis, some forms 79 of diabetes, and Reynaud's syndrome; and transplant rejection disorders such as GVHD and allograft rejection; chronic glomerulonephritis; inflammatory bowel diseases such as chronic matory bowel e (CIBD), Crohn's disease, ulcerative colitis, and necrotizing enterocolitis; inflammatory dermatoses such as contact dermatitis, atopic dermatitis, psoriasis, 5 or urticaria; fever and as due to infection; central or peripheral nervous system inflammatory disorders such as meningitis, encephalitis, and brain or spinal cord injury due to minor trauma; Sjogren's syndrome; diseases involving leukocyte diapedesis; alcoholic hepatitis; bacterial pneumonia; antigen-antibody complex mediated diseases; hypovolemic shock; Type I diabetes mellitus; acute and d hypersensitivity; disease states due to 10 leukocyte dyscrasia and metastasis; thermal injury; granulocyte transfusion-associated syndromes; and cytokine-induced toxicity.
Methods of using compounds of the invention also include ng cancer selected from breast, ovary, cervix, prostate, testis, genitourinary tract, esophagus, larynx, glioblastoma, neuroblastoma, stomach, skin, keratoacanthoma, lung, moid carcinoma, 15 large cell carcinoma, non-small cell lung carcinoma (NSCLC), small cell carcinoma, lung adenocarcinoma, bone, colon, adenoma, pancreas, adenocarcinoma, thyroid, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma, bladder carcinoma, liver carcinoma and biliary passages, kidney carcinoma, pancreatic, myeloid disorders, lymphoma, hairy cells, buccal cavity, naso-pharyngeal, pharynx, lip, 20 tongue, mouth, small intestine, colon-rectum, large intestine, rectum, brain and central nervous , Hodgkin’s, leukemia, bronchus, thyroid, liver and intrahepatic bile duct, hepatocellular, gastric, glioma/glioblastoma, endometrial, melanoma, kidney and renal pelvis, urinary bladder, e , uterine cervix, multiple myeloma, acute enous ia, chronic myelogenous leukemia, cytic leukemia, chronic lymphoid leukemia 25 (CLL), myeloid leukemia, oral cavity and pharynx, non-Hodgkin lymphoma, melanoma, and villous colon a.
The s of using compounds of the invention can have y in treating subjects who are or can be t to reperfusion injury, i.e., injury resulting from situations in which a tissue or organ experiences a period of ia followed by reperfusion. The term 30 "ischemia" refers to localized tissue anemia due to obstruction of the inflow of al blood.
Transient ischemia followed by reperfusion characteristically results in phil activation and transmigration through the endothelium of the blood vessels in the affected area.
Accumulation of activated neutrophils in turn results in generation of reactive oxygen metabolites, which damage components of the involved tissue or organ. This phenomenon of 80 "reperfusion injury" is commonly associated with conditions such as vascular stroke (including global and focal ischemia), hemorrhagic shock, myocardial ischemia or infarction, organ transplantation, and cerebral vasospasm. To illustrate, reperfusion injury occurs at the termination of cardiac bypass procedures or during cardiac arrest when the heart, once 5 ted from receiving blood, begins to reperfuse. It is expected that inhibition of Btk activity may result in reduced amounts of reperfusion injury in such situations.
PHARMACEUTICAL ATIONS In order to use a nd of this invention for the therapeutic ent of mammals including humans, it is normally formulated in accordance with standard pharmaceutical 10 practice as a pharmaceutical composition. According to this aspect of the invention there is provided a pharmaceutical composition comprising a nd of this invention in association with a pharmaceutically acceptable diluent or carrier.
A typical formulation is prepared by mixing a compound of the present invention and a carrier, diluent or excipient. Suitable carriers, diluents and excipients are well known to 15 those skilled in the art and include materials such as carbohydrates, waxes, water soluble and/or swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water and the like. The particular carrier, diluent or ent used will depend upon the means and purpose for which the compound of the present invention is being applied.
Solvents are generally ed based on solvents recognized by persons skilled in the art as 20 safe (GRAS) to be administered to a mammal. In general, safe solvents are xic aqueous solvents such as water and other non-toxic solvents that are soluble or miscible in water. Suitable aqueous solvents include water, ethanol, ene glycol, hylene s (e.g., PEG 400, PEG 300), etc. and es thereof. The formulations may also include one or more buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, 25 emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present invention or pharmaceutical composition f) or aid in the manufacturing of the pharmaceutical product (i.e., medicament). 30 The formulations may be prepared using conventional dissolution and mixing procedures. For example, the bulk drug substance (i.e., compound of the present invention or ized form of the nd (e.g., complex with a extrin tive or other known complexation agent) is dissolved in a suitable solvent in the presence of one or more of the 81 ents described above. The compound of the present invention is typically formulated into pharmaceutical dosage forms to provide an easily controllable dosage of the drug and to enable patient compliance with the prescribed regimen.
The ceutical composition (or formulation) for application may be packaged in 5 a variety of ways depending upon the method used for stering the drug. Generally, an article for distribution includes a container having deposited therein the pharmaceutical formulation in an appropriate form. le containers are well known to those skilled in the art and e materials such as bottles (plastic and glass), sachets, ampoules, plastic bags, metal cylinders, and the like. The container may also include a tamper-proof lage to 10 prevent indiscreet access to the contents of the package. In addition, the container has ted thereon a label that describes the contents of the container. The label may also include appropriate warnings.
Pharmaceutical formulations of the compounds of the present invention may be prepared for various routes and types of administration. For example, a compound of 15 Formula I having the desired degree of purity may optionally be mixed with pharmaceutically acceptable diluents, carriers, excipients or stabilizers (Remington's Pharmaceutical Sciences (1980) 16th edition, Osol, A. Ed.), in the form of a lyophilized formulation, milled powder, or an aqueous solution. Formulation may be conducted by mixing at ambient temperature at the appropriate pH, and at the desired degree of purity, with physiologically able carriers, 20 i.e., carriers that are xic to recipients at the dosages and trations employed. The pH of the formulation depends mainly on the particular use and the concentration of compound, but may range from about 3 to about 8. ation in an acetate buffer at pH 5 is a suitable embodiment.
The compound ordinarily can be stored as a solid composition, a lyophilized 25 formulation or as an aqueous solution.
The ceutical compositions of the invention will be ated, dosed and administered in a fashion, i.e., amounts, concentrations, schedules, course, es and route of administration, consistent with good medical practice. Factors for consideration in this context include the particular er being treated, the particular mammal being treated, the 30 clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners. The "therapeutically effective amount" of the compound to be administered will be ed by such considerations, and is the minimum amount necessary to ameliorate, or treat the hyperproliferative disorder. 82 As a l proposition, the initial pharmaceutically effective amount of the inhibitor administered parenterally per dose will be in the range of about 0.01-100 mg/kg, namely about 0.1 to 20 mg/kg of patient body weight per day, with the l initial range of compound used being 0.3 to 15 mg/kg/day. 5 Acceptable diluents, carriers, excipients and stabilizers are ic to ents at the dosages and trations employed, and include buffers such as phosphate, citrate and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as 10 methyl or propyl paraben; catechol; resorcinol; exanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; ns, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides and other carbohydrates including glucose, mannose, or dextrins; chelating 15 agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g., Zn-protein complexes); and/or non-ionic tants such as TWEEN, PLURONICS or polyethylene glycol (PEG). The active pharmaceutical ingredients may also be entrapped in microcapsules prepared, for example, by vation techniques or by interfacial rization, for e, 20 hydroxymethylcellulose or gelatin-microcapsules and poly-(methylmethacylate) microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980). 25 Sustained-release preparations of compounds of Formula I may be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing a compound of a I, which es are in the form of shaped es, e.g., films, or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate), or poly(vinyl 30 l)), ctides (US 3773919), copolymers of L-glutamic acid and gamma-ethyl-L- glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOT (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate) and poly-D-(-)hydroxybutyric acid. 83 The formulations include those le for the administration routes detailed herein.
The formulations may conveniently be ted in unit dosage form and may be prepared by any of the s well known in the art of pharmacy. Techniques and formulations generally are found in Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, 5 PA). Such s include the step of bringing into association the active ingredient with the carrier which constitutes one or more accessory ingredients. In general the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, g the product. 10 Formulations of a compound of a I suitable for oral administration may be prepared as discrete units such as pills, es, cachets or tablets each containing a predetermined amount of a nd of a I. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, ally mixed with a binder, lubricant, inert diluent, preservative, 15 surface active or dispersing agent. Molded tablets may be made by molding in a le machine a mixture of the powdered active ingredient moistened with an inert liquid t.
The tablets may optionally be coated or scored and optionally are formulated so as to provide slow or controlled release of the active ingredient therefrom. Tablets, troches, lozenges, aqueous or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, 20 e.g., n capsules, syrups or elixirs may be prepared for oral use. Formulations of compounds of Formula I intended for oral use may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents including sweetening agents, ing agents, coloring agents and preserving agents, in order to provide a palatable preparation. Tablets containing 25 the active ingredient in ure with non-toxic pharmaceutically acceptable excipient which are suitable for manufacture of tablets are acceptable. These excipients may be, for example, inert diluents, such as m or sodium carbonate, lactose, calcium or sodium phosphate; granulating and disintegrating agents, such as maize starch, or alginic acid; binding agents, such as starch, gelatin or acacia; and lubricating agents, such as magnesium 30 stearate, stearic acid or talc. Tablets may be ed or may be coated by known techniques including ncapsulation to delay disintegration and adsorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate alone or with a wax may be employed. 84 For treatment of the eye or other external s, e.g., mouth and skin, the formulations are preferably applied as a l ointment or cream containing the active ingredient(s) in an amount of, for example, 0.075 to 20% w/w. When formulated in an ointment, the active ingredients may be employed with either a paraffinic or a water-miscible 5 ointment base. atively, the active ingredients may be formulated in a cream with an oil-in-water cream base. If desired, the aqueous phase of the cream base may include a dric alcohol, i.e., an alcohol having two or more yl groups such as propylene glycol, butane 1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol (including PEG 400) and mixtures thereof. The topical formulations may desirably include a compound 10 which enhances absorption or penetration of the active ient through the skin or other affected areas. Examples of such dermal penetration enhancers include dimethyl sulfoxide and related analogs. The oily phase of the emulsions of this invention may be constituted from known ingredients in a known manner. While the phase may comprise merely an emulsifier, it bly comprises a mixture of at least one emulsifier with a fat or an oil or 15 with both a fat and an oil. Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier which acts as a stabilizer. It is also preferred to include both an oil and a fat. Together, the emulsifier(s) with or without stabilizer(s) make up the so-called emulsifying wax, and the wax together with the oil and fat make up the so-called emulsifying ointment base which forms the oily dispersed phase of the cream formulations. Emulsifiers 20 and emulsion stabilizers suitable for use in the formulation of the invention include Tween® 60, Span® 80, cetostearyl alcohol, benzyl l, myristyl alcohol, glyceryl mono-stearate and sodium lauryl sulfate.
Aqueous sions of a I compounds contain the active materials in ure with excipients suitable for the cture of aqueous suspensions. Such 25 excipients include a suspending agent, such as sodium carboxymethylcellulose, croscarmellose, ne, methylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia, and sing or wetting agents such as a naturally occurring phosphatide (e.g., lecithin), a condensation product of an alkylene oxide with a fatty acid (e.g., polyoxyethylene stearate), a condensation product of ethylene 30 oxide with a long chain aliphatic alcohol (e.g., ecaethyleneoxycetanol), a condensation product of ethylene oxide with a partial ester derived from a fatty acid and a l anhydride (e.g., polyoxyethylene sorbitan monooleate). The aqueous suspension may also contain one or more preservatives such as ethyl or n-propyl p-hydroxybenzoate, one or more 85 ng agents, one or more flavoring agents and one or more sweetening agents, such as sucrose or saccharin.
The ceutical compositions of compounds of Formula I may be in the form of a e injectable preparation, such as a sterile injectable aqueous or oleaginous suspension. 5 This suspension may be formulated ing to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, such as a solution in tanediol or prepared as a lyophilized powder. Among the acceptable vehicles and solvents that may be 10 employed are water, 's solution and isotonic sodium de solution. In addition, sterile fixed oils may conventionally be employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides.
In addition, fatty acids such as oleic acid may likewise be used in the preparation of injectables. 15 The amount of active ient that may be combined with the carrier material to produce a single dosage form will vary ing upon the host treated and the particular mode of administration. For example, a time-release formulation intended for oral administration to humans may n approximately 1 to 1000 mg of active material compounded with an appropriate and convenient amount of carrier al which may vary 20 from about 5 to about 95% of the total compositions (weight:weight). The pharmaceutical composition can be prepared to provide easily measurable amounts for administration. For example, an s solution intended for intravenous infusion may contain from about 3 to 500 µg of the active ingredient per milliliter of on in order that infusion of a suitable volume at a rate of about 30 mL/hr can occur. 25 Formulations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include ding agents and thickening agents. 30 Formulations suitable for topical administration to the eye also include eye drops wherein the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active ingredient. The active ingredient is preferably t in such formulations in a concentration of about 0.5 to 20% w/w, for example about 0.5 to 10% w/w, for example about 1.5% w/w. 86 Formulations suitable for topical administration in the mouth include lozenges comprising the active ingredient in a flavored basis, y sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert basis such as gelatin and glycerin, or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid 5 carrier.
Formulations for rectal administration may be presented as a suppository with a suitable base comprising for example cocoa butter or a salicylate.
Formulations suitable for intrapulmonary or nasal administration have a particle size for e in the range of 0.1 to 500 s (including particle sizes in a range between 10 0.1 and 500 microns in increments microns such as 0.5, 1, 30 s, 35 microns, etc.), which is stered by rapid tion through the nasal passage or by inhalation through the mouth so as to reach the alveolar sacs. Suitable formulations include aqueous or oily solutions of the active ingredient. Formulations suitable for aerosol or dry powder administration may be prepared according to conventional s and may be delivered 15 with other therapeutic agents such as compounds heretofore used in the treatment or prophylaxis disorders as described below.
Formulations suitable for vaginal stration may be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the active ingredient such carriers as are known in the art to be appropriate. 20 The formulations may be packaged in unit-dose or dose containers, for example sealed es and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water, for injection immediately prior to use. Extemporaneous injection solutions and suspensions are prepared from sterile powders, granules and tablets of the kind previously described. Preferred unit 25 dosage formulations are those containing a daily dose or unit daily sub-dose, as herein above recited, or an riate fraction thereof, of the active ingredient.
The invention further provides veterinary compositions comprising at least one active ient as above defined together with a veterinary carrier therefore. Veterinary carriers are materials useful for the purpose of stering the ition and may be solid, liquid 30 or gaseous materials which are otherwise inert or acceptable in the veterinary art and are ible with the active ingredient. These veterinary compositions may be administered parenterally, orally or by any other desired route.
COMBINATION THERAPY 87 The compounds of Formula I may be ed alone or in combination with other therapeutic agents for the treatment of a disease or disorder described herein, such as inflammation or a hyperproliferative disorder (e.g., cancer). In certain embodiments, a compound of Formula I is combined in a pharmaceutical combination formulation, or dosing 5 regimen as combination therapy, with an onal, second eutic compound that has anti-inflammatory or yperproliferative properties or that is useful for treating an inflammation, immune-response disorder, or hyperproliferative er (e.g., cancer). The additional therapeutic may be an anti-inflammatory agent, an immunomodulatory agent, chemotherapeutic agent, an apoptosis-enhancer, a ropic factor, an agent for treating 10 cardiovascular disease, an agent for treating liver disease, an anti-viral agent, an agent for treating blood disorders, an agent for treating diabetes, and an agent for treating immunodeficiency disorders. The second therapeutic agent may be an NSAID antiinflammatory agent. The second therapeutic agent may be a chemotherapeutic agent. The second compound of the pharmaceutical combination formulation or dosing regimen 15 preferably has complementary activities to the compound of a I such that they do not adversely affect each other. Such compounds are ly present in combination in amounts that are effective for the purpose intended. In one embodiment, a composition of this invention comprises a compound of a I, or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt thereof, in combination with a therapeutic agent such as an 20 NSAID.
The ation therapy may be administered as a simultaneous or sequential regimen. When administered sequentially, the combination may be administered in two or more administrations. The combined administration includes coadministration, using separate formulations or a single pharmaceutical formulation, and consecutive administration 25 in either order, wherein preferably there is a time period while both (or all) active agents aneously exert their biological activities.
Suitable dosages for any of the above coadministered agents are those presently used and may be lowered due to the combined action (synergy) of the newly identified agent and other therapeutic agents or treatments. 30 The combination therapy may provide "synergy" and prove "synergistic", i.e., the effect achieved when the active ingredients used er is greater than the sum of the effects that s from using the compounds separately. A synergistic effect may be attained when the active ingredients are: (1) co-formulated and administered or delivered simultaneously in a combined, unit dosage ation; (2) delivered by alternation or in 88 parallel as separate formulations; or (3) by some other regimen. When delivered in alternation therapy, a synergistic effect may be attained when the compounds are stered or delivered sequentially, e.g., by different injections in separate syringes, separate pills or capsules, or separate infusions. In general, during alternation y, an 5 effective dosage of each active ingredient is stered sequentially, i.e., serially, whereas in combination therapy, effective dosages of two or more active ingredients are administered together.
In a particular embodiment of therapy, a compound of Formula I, or a stereoisomer, tautomer, solvate, or ceutically acceptable salt thereof, may be combined with other 10 therapeutic, hormonal or antibody agents such as those described herein, as well as combined with surgical y and radiotherapy. Combination ies described herein thus comprise the administration of at least one compound of Formula I, or a stereoisomer, tautomer, solvate, or ceutically acceptable salt f, and the use of at least one other cancer treatment method. The amounts of the compound(s) of Formula I and the other 15 pharmaceutically active therapeutic agent(s) and the relative timings of administration will be selected in order to achieve the desired combined eutic effect.
METABOLITES OF COMPOUNDS OF FORMULA I Also described herein are the in vivo metabolic products of Formula I described herein. Such products may result for example from the oxidation, reduction, hydrolysis, 20 amidation, deamidation, esterification, deesterification, enzymatic ge, and the like, of the administered compound. ingly,also described herein are metabolites of compounds of Formula I, including compounds produced by a process comprising contacting a compound of this invention with a mammal for a period of time sufficient to yield a metabolic product thereof. 25 Metabolite ts typically are identified by preparing a radiolabelled (e.g., 14C or 3H) isotope of a compound of the invention, administering it parenterally in a detectable dose (e.g., greater than about 0.5 mg/kg) to an animal such as rat, mouse, guinea pig, monkey, or to man, allowing sufficient time for metabolism to occur (typically about 30 seconds to 30 hours) and ing its conversion products from the urine, blood or other biological samples. 30 These products are easily isolated since they are d (others are isolated by the use of antibodies capable of binding epitopes surviving in the metabolite). The metabolite structures are ined in conventional fashion, e.g., by MS, LC/MS or NMR analysis. In general, analysis of metabolites is done in the same way as conventional drug metabolism 89 s well known to those skilled in the art. The metabolite products, so long as they are not ise found in vivo, are useful in diagnostic assays for therapeutic dosing of the compounds of the invention.
ARTICLES OF MANUFACTURE 5 In another embodiment of the invention, an article of manufacture, or "kit", containing materials useful for the treatment of the diseases and disorders described above is provided.
In one embodiment, the kit comprises a container comprising a compound of Formula I, or a stereoisomer, er, solvate, or pharmaceutically acceptable salt thereof. The kit may further comprise a label or package insert on or associated with the container. The term 10 "package insert" is used to refer to ctions customarily included in commercial packages of therapeutic products, that contain information about the indications, usage, dosage, stration, contraindications and/or warnings concerning the use of such therapeutic products. Suitable containers e, for example, bottles, vials, syringes, blister pack, etc.
The container may be formed from a variety of materials such as glass or plastic. The 15 container may hold a compound of Formula I or a formulation thereof which is effective for treating the condition and may have a sterile access port (for example, the container may be an intravenous solution bag or a vial having a stopper able by a hypodermic injection ). At least one active agent in the composition is a compound of a I. The label or package insert indicates that the composition is used for treating the ion of choice, 20 such as cancer. In addition, the label or package insert may indicate that the t to be treated is one having a disorder such as a hyperproliferative disorder, neurodegeneration, cardiac hypertrophy, pain, migraine or a neurotraumatic disease or event. In one embodiment, the label or e inserts indicates that the composition comprising a compound of Formula I can be used to treat a disorder resulting from abnormal cell growth. The label or 25 package insert may also indicate that the composition can be used to treat other disorders.
Alternatively, or additionally, the article of manufacture may further comprise a second container comprising a pharmaceutically acceptable , such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution and dextrose solution. It may further include other als desirable from a commercial and user standpoint, including 30 other buffers, diluents, filters, needles, and syringes.
The kit may further comprise directions for the administration of the compound of Formula I and, if present, the second pharmaceutical formulation. For example, if the kit comprises a first composition sing a compound of Formula I and a second 90 ceutical formulation, the kit may further comprise directions for the simultaneous, sequential or separate administration of the first and second pharmaceutical compositions to a patient in need thereof.
In another embodiment, the kits are suitable for the delivery of solid oral forms of a 5 compound of Formula I, such as tablets or capsules. Such a kit preferably es a number of unit dosages. Such kits can include a card having the dosages oriented in the order of their intended use. An example of such a kit is a "blister pack". Blister packs are well known in the ing industry and are widely used for ing pharmaceutical unit dosage forms.
If desired, a memory aid can be ed, for example in the form of numbers, letters, or 10 other gs or with a calendar insert, designating the days in the treatment le in which the s can be administered.
According to one embodiment, a kit may comprise (a) a first container with a compound of Formula I contained therein; and optionally (b) a second container with a second pharmaceutical formulation contained n, wherein the second pharmaceutical 15 formulation comprises a second compound with anti-hyperproliferative activity.
Alternatively, or additionally, the kit may further comprise a third container comprising a pharmaceutically-acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution and dextrose solution. It may further include other als desirable from a commercial and user standpoint, including other buffers, 20 diluents, filters, needles, and syringes.
In certain other embodiments wherein the kit comprises a composition of Formula I and a second therapeutic agent, the kit may comprise a container for containing the separate compositions such as a divided bottle or a divided foil packet, however, the separate compositions may also be contained within a , undivided container. Typically, the kit 25 comprises directions for the administration of the separate components. The kit form is particularly advantageous when the separate components are preferably administered in different dosage forms (e.g., oral and parenteral), are administered at different dosage intervals, or when ion of the individual components of the combination is desired by the prescribing physician. 30 PREPARATION OF FORMULA I COMPOUNDS Compounds of Formula I may be synthesized by synthetic routes that include processes analogous to those well-known in the chemical arts, particularly in light of the description ned herein, and those for other heterocycles bed in: Comprehensive 91 Heterocyclic Chemistry II, Editors Katritzky and Rees, Elsevier, 1997, e.g. Volume 3; Liebigs Annalen der Chemie, (9):1910-16, (1985); Helvetica Chimica Acta, 41:1052-60, (1958); Arzneimittel-Forschung, 40(12):1328-31, (1990), each of which are expressly incorporated by reference. Starting materials are generally ble from commercial 5 s such as Aldrich als (Milwaukee, WI) or are readily prepared using methods well known to those skilled in the art (e.g., prepared by methods generally described in Louis F. Fieser and Mary Fieser, Reagents for Organic Synthesis, v. 1-23, Wiley, N.Y. (1967-2006 ed.), or Beilsteins Handbuch der organischen Chemie, 4, Aufl. ed. Springer-Verlag, Berlin, including supplements (also available via the Beilstein online database). 10 Synthetic chemistry transformations and protecting group methodologies ction and deprotection) useful in synthesizing Formula I compounds and necessary reagents and intermediates are known in the art and include, for example, those described in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T. W. Greene and P.
G .M. Wuts, tive Groups in Organic sis, 3rd Ed., John Wiley and Sons (1999); 15 and L. Paquette, ed., Encyclopedia of ts for Organic Synthesis, John Wiley and Sons (1995) and subsequent editions thereof.
Compounds of Formula I may be prepared singly or as compound libraries comprising at least 2, for example 5 to 1,000 compounds, or 10 to 100 nds. Libraries of compounds of Formula I may be prepared by a combinatorial ‘split and mix’ approach or 20 by multiple parallel syntheses using either solution phase or solid phase chemistry, by ures known to those skilled in the art. Thus ing to a further aspect of the invention there is provided a nd library comprising at least 2 compounds, or pharmaceutically acceptable salts thereof.
The Figures and Examples provide exemplary methods for preparing Formula I 25 nds. Those d in the art will appreciate that other synthetic routes may be used to synthesize the Formula I compounds. Although specific starting materials and reagents are depicted and discussed in the Figures and Examples, other starting als and reagents can be easily substituted to provide a variety of derivatives and/or reaction conditions. In addition, many of the exemplary compounds ed by the bed s can be 30 further modified in light of this disclosure using conventional chemistry well known to those skilled in the art.
In preparing compounds of Formulas I, protection of remote functionality (e.g., primary or secondary amine) of intermediates may be necessary. The need for such protection will vary depending on the nature of the remote functionality and the conditions of 92 the preparation methods. Suitable amino-protecting groups include , trifluoroacetyl, tbutoxycarbonyl (BOC), benzyloxycarbonyl (CBz) and 9-fluorenylmethyleneoxycarbonyl (Fmoc). The need for such protection is readily determined by one skilled in the art. For a l description of protecting groups and their use, see T. W. Greene, Protective Groups in 5 c Synthesis, John Wiley & Sons, New York, 1991.
Experimental procedures, intermediates and reagents useful for useful for the preparation of Formula I compounds may be found in US Ser. No. 13/102720, ONE AND AZA-PYRIDONE COMPOUNDS AND METHODS OF USE”, filed 6 May 2011, which is orated by reference in its entirety. 10 Figures 1-24 describe the synthesis of exemplary embodiments of Formula I compounds 101-124, more fully described in Examples 4, and may be useful for the preparation of other Formula I compounds.
GENERAL PREPARATIVE PROCEDURES General Procedure: Suzuki Coupling R5 O O R5 HN HN B B O O O O Y2 Y2 O N N B X Y1 R6 Y1 R6 O X = Br, Cl X = Br, Cl A-1 B-2 N R4 O O N R4 O B B N Br O O N B O O X1 X3 O X1 X3 X2 X2 B-4 A-2 R5 HN B-2 + A-2 N R4 O Y2 Suzuki Reaction or N N Y1 R6 A-1 + B-4 O X1 X3 X2 15 A-3 The Suzuki-type coupling reaction is useful to form carbon-carbon bonds to attach the rings of Formula I compounds and intermediates such as A-3 (Suzuki (1991) Pure Appl.
Chem. 63:419-422; Miyaura and Suzuki (1979) Chem. Reviews 95(7):2457-2483; Suzuki 93 (1999) J. Organometal. Chem. 576:147-168). Suzuki coupling is a palladium mediated cross coupling reaction of a heteroarylhalide, such as B-2 or B-4, with a boronic acid such as A-1 or A-2. For example, B-2 may be combined with about 1.5 equivalents of 4,4,4',4',5,5,5',5'- octamethyl-2,2'-bi(1,3,2-dioxaborolane), and dissolved in about 3 equivalents of sodium 5 carbonate as a 1 molar solution in water and an equal volume of acetonitrile. A catalytic amount, or more, of a low valent palladium reagent, such as bis(triphenylphosphine)palladium(II) dichloride, is added. In some cases potassium acetate is used in place of sodium carbonate to adjust the pH of the aqueous layer. The reaction is then heated to about 140-150 °C under re in a microwave reactor (Biotage AB, Uppsala, 10 Sweden) for 10 to 30 minutes. The contents are ted with ethyl acetate, or another organic solvent. After evaporation of the organic layer the boron ester A-1 may be purified on silica or by reverse phase HPLC. tuents are as defined, or protected forms or precursors f. Likewise, bromide intermediate B-4 can be lated to give A-2.
Suzuki ng of B-2 and A-2, or of A-1 and B-4, gives Formula I compound or 15 intermediate A-3. Boronic ester (or acid) (1.5 eq) A-1 or A-2, and a ium catalyst such as bis(triphenylphosphine)palladium(II) chloride (0.05 eq) is added to a e of halo intermediate (1 eq) B-2 or B-4 in acetonitrile and 1 M of sodium carbonate aqueous solution (equal volume as acetonitrile). The reaction mixture is heated to about 150 oC in a microwave for about 15 min. LC/MS indicates when the reaction is complete. Water is 20 added to the mixture, and the precipitated t is ed and purified by HPLC to yield the product A-3. Substituents R1’, R2’, R4’ may be R1, R2, R4 as defined, or protected forms or precursors thereof.
A variety of ium catalysts can be used during the Suzuki coupling step. Various low valent, Pd(II) and Pd(0) catalysts may be used in the Suzuki coupling reaction, including 25 PdCl2(PPh3)2, Pd(t-Bu)3, PdCl2 dppf CH2Cl2, Pd(PPh3)4, Pd(OAc)/PPh3, Cl2Pd[(Pet3)]2, Pd(DIPHOS)2, Cl2Pd(Bipy), [PdCl(Ph2PCH2PPh2)]2, Cl2Pd[P(o-tol)3]2, Pd2(dba)3/P(o-tol)3, Pd2(dba)/P(furyl)3, Cl2Pd[P(furyl)3]2, Cl2Pd(PMePh2)2, Cl2Pd[P(4-F-Ph)3]2, Cl2Pd[P(C6F6)3]2, P(2-COOH-Ph)(Ph)2]2, Cl2Pd[P(4-COOH-Ph)(Ph)2]2, and encapsulated catalysts Pd EnCat™ 30, Pd EnCat™ TPP30, and Pd(II)EnCat™ BINAP30 (US 2004/0254066). 30 General Procedure: Buchwald reaction 94 The Buchwald reaction is useful to aminate 6-bromo ediates B-1 (Wolf and Buchwald (2004) Org. Synth Coll. Vol. 10:423; Paul et al (1994) Jour. Amer. Chem. Soc. 116:5969-5970). To a solution of halo intermediate B-1 in DMF is added the riate 5 amine R5-NH2 (200 mol %), Cs2CO3 (50 mol%), Pd2(dba)3 (5 mol%), and 4,5- bis(diphenylphosphino)-9,9-dimethylxanthene (Xantphos, CAS Reg. No. 1612658, 10 mol%). The reaction is heated to about 110 °C under pressure in a microwave reactor (Biotage AB, Uppsala, Sweden) for about 30 min. The resulting solution is concentrated in vacuo to give B-2. Other palladium catalysts and phosphine ligands may be useful. 10 N-Heteroaryl amide intermediates B-4 can also be prepared under Buchwald conditions with cyclic amide intermediates (R7) such as 3,4,6,7,8,9-hexahydropyrazino[1,2- l-1(2H)-one 101e and heteroaryl dibromides B-3.
METHODS OF SEPARATION 15 In the methods of preparing a I compounds, it may be advantageous to separate reaction products from one another and/or from starting materials. The desired products of each step or series of steps is separated and/or purified to the desired degree of neity by the ques common in the art. Typically such separations involve multiphase extraction, crystallization from a t or solvent mixture, distillation, 20 sublimation, or chromatography. Chromatography can involve any number of methods including, for example: reverse-phase and normal phase; size exclusion; ion ge; high, medium and low pressure liquid chromatography methods and apparatus; small scale 95 analytical; simulated moving bed (SMB) and ative thin or thick layer chromatography, as well as techniques of small scale thin layer and flash chromatography.
Another class of separation s involves treatment of a mixture with a reagent selected to bind to or render otherwise separable a desired product, unreacted starting 5 material, reaction by product, or the like. Such reagents e adsorbents or absorbents such as activated carbon, molecular sieves, ion ge media, or the like. atively, the reagents can be acids in the case of a basic al, bases in the case of an acidic material, binding reagents such as antibodies, binding proteins, selective chelators such as crown , liquid/liquid ion extraction reagents (LIX), or the like. Selection of appropriate methods of 10 separation depends on the nature of the materials involved, such as, boiling point and molecular weight in distillation and sublimation, presence or absence of polar onal groups in chromatography, stability of materials in acidic and basic media in multiphase extraction, and the like.
Diastereomeric mixtures can be separated into their individual diastereomers on the 15 basis of their physical chemical differences by methods well known to those skilled in the art, such as by chromatography and/or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active nd (e.g., chiral auxiliary such as a chiral alcohol or Mosher’s acid chloride), separating the diastereomers and converting (e.g., hydrolyzing) the 20 dual diastereoisomers to the corresponding pure enantiomers. Also, some of the nds of the present invention may be atropisomers (e.g., substituted biaryls) and are considered as part of this invention. Enantiomers can also be separated by use of a chiral HPLC column.
A single stereoisomer, e.g., an enantiomer, substantially free of its stereoisomer may 25 be obtained by resolution of the racemic mixture using a method such as formation of diastereomers using lly active resolving agents (Eliel, E. and Wilen, S. ochemistry of Organic Compounds,” John Wiley & Sons, Inc., New York, 1994; Lochmuller, C. H., (1975) J. Chromatogr., :283-302). Racemic mixtures of chiral compounds of the invention can be separated and isolated by any suitable method, including: 30 (1) formation of ionic, diastereomeric salts with chiral compounds and tion by fractional crystallization or other methods, (2) formation of diastereomeric compounds with chiral derivatizing reagents, separation of the reomers, and conversion to the pure stereoisomers, and (3) separation of the substantially pure or enriched stereoisomers directly 96 under chiral conditions. See: “Drug Stereochemistry, Analytical Methods and cology,” Irving W. Wainer, Ed., Marcel Dekker, Inc., New York (1993).
Under method (1), diastereomeric salts can be formed by reaction of enantiomerically pure chiral bases such as brucine, quinine, ephedrine, strychnine, α-methyl-β- 5 phenylethylamine (amphetamine), and the like with asymmetric compounds bearing acidic onality, such as carboxylic acid and ic acid. The diastereomeric salts may be induced to separate by fractional crystallization or ionic chromatography. For separation of the optical isomers of amino compounds, addition of chiral carboxylic or sulfonic acids, such as rsulfonic acid, tartaric acid, mandelic acid, or lactic acid can result in formation of 10 the diastereomeric salts.
Alternatively, by method (2), the substrate to be resolved is d with one enantiomer of a chiral compound to form a diastereomeric pair (E. and Wilen, S.
“Stereochemistry of Organic Compounds”, John Wiley & Sons, Inc., 1994, p. 322).
Diastereomeric compounds can be formed by reacting asymmetric compounds with 15 enantiomerically pure chiral derivatizing reagents, such as menthyl derivatives, followed by separation of the diastereomers and hydrolysis to yield the pure or enriched enantiomer. A method of determining optical purity involves making chiral esters, such as a menthyl ester, e.g., (-) menthyl chloroformate in the presence of base, or Mosher ester, oxy-α- (trifluoromethyl)phenyl acetate (Jacob III. J. Org. Chem. (1982) 47:4165), of the c 20 mixture, and analyzing the 1H NMR spectrum for the presence of the two atropisomeric enantiomers or diastereomers. Stable diastereomers of atropisomeric compounds can be separated and isolated by normal- and reverse-phase chromatography following methods for separation of atropisomeric naphthyl-isoquinolines (WO 11). By method (3), a c mixture of two enantiomers can be separated by chromatography using a chiral 25 nary phase (“Chiral Liquid Chromatography” (1989) W. J. Lough, Ed., Chapman and Hall, New York; Okamoto, J. Chromatogr., (1990) 513:375-378). Enriched or purified enantiomers can be distinguished by s used to distinguish other chiral molecules with asymmetric carbon atoms, such as optical rotation and circular dichroism.
EXAMPLES 30 Example 101a 2,2,2-Trichloro(4,5,6,7-tetrahydro-1H-indolyl)ethanone 101a 97 NH O Cl3C 101a A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer, condenser and nitrogen inlet was purged with nitrogen and charged with 4,5,6,7-tetrahydro- 1H-indole (3.00 g, 24.8 mmol), trichloroacetyl chloride (13.5 g, 74.4 mmol) and 1,2- 5 dichloroethane (50 mL). The solution was stirred at 85 oC for 2 h. After that time, the reaction mixture was trated under reduced re to afford a 100% yield (6.50 g) of 2,2,2-trichloro(4,5,6,7-tetrahydro-1H-indolyl)ethanone 101a as a black olid: 1H NMR (500 MHz, DMSO-d6) δ 11.94 (s, 1H), 7.05 (s, 1H), 2.62 (t, 2H, J = 6.0 Hz), 2.47 (t, 2H, J = 6.0 Hz), 1.80 (m, 2H), 1.65 (m, 2H); MS (ESI+) m/z 266.0 (M+H) 10 Example 101b Ethyl 4,5,6,7-Tetrahydro-1H-indolecarboxylate 101b NH CO2Et 101b A 100-mL single-neck bottomed flask equipped with a magnetic stirrer and nitrogen inlet was purged with nitrogen and charged with 101a (6.50 g, 24.8 mmol), sodium ethoxide (17.0 mg, 0.25 mmol) and ethanol (40 mL). The solution was d at room 15 temperature for 1 h. After that time, the reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography to afford a 100% yield (4.80 g) of ethyl 4,5,6,7-tetrahydro-1H-indolecarboxylate 101b as a brown solid: mp 70–72 °C; 1H NMR (300 MHz, CDCl 3) δ 9.08 (s, 1H), 6.75 (s, 1H), 4.25 (q, 2H, J = 7.2 Hz), 2.65 (t, 2H, J = 6.0 Hz), 2.56 (t, 2H, J = 6.0 Hz), 1.85 (m, 4H), 1.28 (t, 3H, J = 7.2 Hz); MS (ESI+) m/z 20 194.1 (M+H) Example 101c Ethyl 1-(Cyanomethyl)-4,5,6,7-tetrahydro-1H-indole carboxylate 101c N CN CO2Et 101c 98 A 125-mL single-neck round-bottomed flask equipped with a magnetic stirrer and nitrogen inlet was purged with nitrogen and charged with 101b (5.76 g, 29.8 mmol) and DMF (50 mL). The solution was cooled to 0 °C using an ice bath. NaH (60% sion in mineral oil, 1.43 g, 35.8 mmol) was added. The resulting mixture was stirred at room temperature for 5 1 h. After that time, cetonitrile (1.43 g, 35.8 mmol) was added. The mixture was stirred at room temperature for 14 h. After that time, the reaction mixture was concentrated under reduced pressure and the residue was partitioned between ethyl acetate (150 mL) and water (450 mL). The organic layer was separated, and the s layer was extracted with ethyl e (3 × 150 mL). The combined organic layers were washed with brine, dried over 10 sodium sulfate and concentrated under reduced re. The residue was purified by column chromatography to afford a 55% yield (3.80 g) of ethyl 1-(cyanomethyl)-4,5,6,7-tetrahydro- 1H-indolecarboxylate 101c as a yellow semi-solid: 1H NMR (300 MHz, CDCl3) δ 6.66 (s, 1H), 5.29 (s, 2H), 4.28 (q, 2H, J = 7.2 Hz), 2.62 (t, 2H, J = 6.3 Hz), 2.49 (t, 2H, J = 6.3 Hz), 1.92 (m, 2H), 1.75 (m, 2H), 1.33 (t, 3H, J = 7.2 Hz); MS (ESI+) m/z 233.1 (M+H) 15 Example 101d Ethyl 1-(2-Aminoethyl)-4,5,6,7-tetrahydro-1H-indole carboxylate 101d N NH2 CO2Et 101d A 200-mL Parr reactor bottle was purged with nitrogen and charged with 10% palladium on carbon (50% wet, 1.28 g dry weight), 101c (3.00 g, 12.9 mmol), 12% 20 hydrochloric acid (6.5 mL, 25 mmol), ethyl acetate (60 mL) and ethanol (40 mL). The bottle was ed to a Parr hydrogenator, evacuated, charged with hydrogen gas to a pressure of 50 psi and shaken for 6 h. After this time, the hydrogen was evacuated, and nitrogen was charged into the bottle. diatomaceous earth filter agent (CELITE®, Imerys Minerals California, Inc.) CELITE® 521 (4.0 g) was added, and the mixture was filtered through a pad 25 of CELITE® 521. The filter cake was washed with l (2 x 20 mL), and the combined filtrates were concentrated to dryness under reduced pressure. The residue was partitioned between ethyl acetate (150 mL) and 10% aqueous potassium carbonate (100 mL). The organic layer was separated, and the aqueous layer was ted with ethyl acetate (3 x 75 mL). The combined c layers were dried over sodium e and concentrated under 30 reduced pressure. The residue was triturated with ethanol (5 mL) to afford a 71% yield (1.71 g) of ethyl 1-(2-aminoethyl)-4,5,6,7-tetrahydro-1H-indolecarboxylate 101d as a white 99 solid: mp 102–104 °C; 1H NMR (500 MHz, DMSO-d6) δ 6.61 (s, 1H), 6.22 (br, 2H), 4.15 (m, 4H), 2.77 (m, 2H), 2.59 (t, 2H, J = 6.5 Hz), 2.42 (t, 2H, J = 6.5 Hz), 1.70 (m, 2H), 1.62 (m, 2H), 1.23 (t, 3H, J = 7.0 Hz); MS (APCI+) m/z 237.2 (M+H) Example 101e 3,4,6,7,8,9-Hexahydropyrazino[1,2-a]indol-1(2H)-one 101e N NH O 5 101e A 100-mL -neck bottomed flask equipped with a magnetic stirrer and nitrogen inlet was purged with nitrogen and charged with 101d (1.80 g, 7.63 mmol), sodium de (1.55 g, 22.8 mmol) and ethanol (50 mL). The mixture was stirred at 55 oC for 5 h.
After that time, the reaction mixture was concentrated under reduced re and the residue 10 was partitioned between ethyl acetate (200 mL) and water (100 mL). The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (2 × 100 mL). The combined organic layers were washed with brine, dried over sodium sulfate and concentrated under reduced pressure. The e was purified by column chromatography to afford a 42% yield (605 mg) of 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 101e as a white 15 solid: mp 207–209 °C; 1H NMR (500 MHz, DMSO-d6) δ 7.41 (s, 1H), 6.36 (s, 1H), 3.84 (t, 2H, J = 6.0 Hz), 3.42 (m, 2H), 2.51 (t, 2H, J = 6.0 Hz), 2.42 (t, 2H, J = 6.0 Hz), 1.76 (m, 2H), 1.65 (m, 2H); (APCI+) m/z 191.3 (M+H) Example 101f 3-Bromo(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl)isonicotinaldehyde 101f 20 A 50-mL single-neck round-bottomed flask equipped with a ic stirrer and a reflux condenser was charged with 101e (300 mg, 1.57 mmol), 3,5- dibromoisonicotinaldehyde (2) (517 mg, 1.96 mmol), 4,5-bis(diphenylphosphino)-9,9- dimethylxanthene (XantPhos, 120 mg, 0.2 mmol), tris(dibenzylideneacetone)dipalladium(0) 25 (180 mg, 0.2 mmol), Cs2CO3 (650 mg, 2 mmol), and 1,4-dioxane (8 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100 oC for 6 h. It was then cooled to room 100 temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by flash column chromatography eluting with DCM/MeOH (from 40:1 to 20:1) to afford 101f as a pale yellow solid (350 mg, 40%). MS: [M+H]+ 374.
Example 101g utyl 4-(6-Nitropyridinyl)piperazinecarboxylate 5 101g Boc N N N NO2 101g Into a solution of 5-bromonitropyridine (30 g, 148 mmol) in DMSO (1 L) were added K2CO3 (40 g, 296 mmol) and tert-butyl piperazinecarboxylate (28g, 148 mmol).
The e was stirred at 65 ºC overnight. After cooling down, it was poured into water (2 10 L). The solid precipitated was collected and dried under vacuum. It was then further purified by flash column eluting with 20:1 petroleum ether/ethyl acetate and then with methylene de to give 101g as a yellow solid (17 g, 37%). MS: [M+H]+ 309.
Example 101h tert-Butyl 4-(6-Aminopyridinyl)piperazinecarboxylate 101h Boc N N N NH2 15 101h A 500-mL bottle was purged with nitrogen and charged with tert-butyl 4-(6- nitropyridinyl)piperazinecarboxylate 101g (3.1 g, 10 mmol), 10% palladium on carbon (50% wet, 1.0 g) and ethanol (100 mL). It was evacuated, charged with hydrogen gas, and stirred for 16 h at room ature. The hydrogen was then evacuated and nitrogen was 20 charged into the bottle. The catalyst was removed by filtration through a pad of CELITE® and the filtrate trated under reduced pressure to afford 101h (2.7 g, 97%). MS: [M+H]+ 279 Example 101i tert-Butyl 4-(6-(5-bromomethyloxo-1,2-dihydropyridin- 3-ylamino)pyridineyl)piperazinecarboxylate 101i 101 Boc N N N NH O N Br 101i A 100-mL single-neck round-bottomed flask ed with a magnetic stirrer and reflux condenser was charged with oxane (50 mL), 101h (1.3 g, 4.7 mmol), 3,5- dibromomethylpyridin-2(1H)-one (1.24 g, 4.7 mmol), and cesium carbonate (3.8 g, 12 5 mmol). After bubbling nitrogen through the resulting mixture for 30 minutes, os (272 mg, 0.47 mmol) and ibenzylideneacetone)dipalladium(0) (430 mg, 0.47 mmol) were added, and the reaction mixture was heated at reflux for 3 h. After this time the reaction was cooled to room temperature, partitioned between ethyl acetate (100 mL) and water (100 mL), and filtered. The aqueous layer was separated and extracted with ethyl acetate (50 mL × 2). 10 The organic layers were combined, washed with brine (50 mL), and dried over sodium sulfate.
The drying agent was removed by filtration and the filtrate was concentrated under d pressure. The residue was purified on flash column eluting with 50:1 methylene chloride/methanol to afford 101i (1.3 g, 59%). MS: [M+H]+ 464.
Example 101j 5-Bromomethyl(5-(piperazinyl)pyridin 15 ylamino)pyridin-2(1H)-one 101j HN N N NH O N Br 101j A mixture of 101i (3.6 g, 7.8 mmol) and 4.0 M HCl/dioxane (10 mL) was stirred for 5 h at room temperature. It was then concentrated at reduced pressure. The residue was basified with aqueous 1.0M NaOH and extracted with methylene chloride. The ed organic 20 layers were washed with water and concentrated under reduced pressure to give 101j (2.46 g, 87%). MS: [M+H]+ 364.
Example 101k 5-Bromomethyl(5-(4-(oxetanyl)piperazinyl)pyridin- 2-ylamino)pyridin-2(1H)-one 101k 102 O N N N NH O N Br 101k A mixture of 101j (2.75 g, 7.5 mmol), oxetanone (1.6 g, 22.7 mmol), NaBH3CN (4.75 g, 22.5 mmol), and zinc chloride (3 g, 22.7 mmol) in methanol (125 mL) was d for 5 hours at 50 ºC. The mixture was added to water and extracted with methylene chloride for 5 three times. The organic layers were concentrated under reduced pressure. The residue was purified by column tography eluting with 25:1 methylene chloride/methanol to give 101k (1.92 g, 61%). MS: [M+H]+ 420. 1H NMR (500 MHz, DMSO) δ 8.58 (d, J = 2.5, 1H), 8.55 (s, 1H), 7.94 (d, J = 3, 1H), 7.54 (d, J = 2.5, 1H), 7.39 (dd, J = 3, 1H), 7.25 (d, J = 4, 1H), 4.56 (t, J = 6.5, 2H), 4.46 (t, J = 6.5, 2H), 3.50 (s, 3H), 3.43 (m, 1 H), 3.01 (m, 4H), 2.40 (m, 10 4H).
Example 101l 1-Methyl(5-(4-(oxetanyl)piperazinyl)pyridin o)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 101l A 500-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 101k (10.5 g, 25 mmol), Pin2B2 (15.6 g, 2.5 eq., 62 mmol), 15 Pd2(dba)3 (1.14 g, 0.05 eq., 1.25 mmol), X-phos (1.16 g, 0.1 eq., 2.5 mmol), AcOK (7.35 g, 3 eq., 75 mmol) and dioxane (150 mL). After three cycles of vacuum/argon flush, the e was heated to 65 ºC for 14 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the ing residue was washed by PE/EA=3/1 (80 mL) to afford 101l as a yellow solid (10.5 g, 94%). MS: [M+H]+ 468. 20 Example 101m 3-(1-methyl(5-(4-(oxetanyl)piperazinyl)pyridin ylamino)oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)isonicotinaldehyde 101m A sealed tube was charged with 101f (200 mg, 0.53 mmol), 101l (250 mg, 0.53 mmol), PdCl2(dppf) (42 mg, 0.05mmol), K3PO4 (210 mg, 1.0 mmol), and NaOAc (85 mg, 1.0 mmol) 25 in acetonitrile/H2O (8 mL/1mL). After three cycles of vacuum/argon flush, the mixture was heated at 100 oC in a sealed tube for 4 h. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified on reverse phase flash eluting with 20:1 DCM/MeOH to afford 101m (135 mg, 40%). LCMS: [M+H]+ 635 . 103 Example 101 2-(4-(hydroxymethyl)(1-methyl(5-(4-(oxetanyl)piperazin yl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)-3,4,6,7,8,9- dropyrazino[1,2-a]indol-1(2H)-one 101 A mixture of 101m (135 mg, 0.21 mmol) and NaBH4 (20 mg, 0.5 mmol) in MeOH (5 5 mL) was stirred at 0 oC for 0.5 h. The mixture was quenched with water and the residue was extracted with EtOAc (5 mL X 2). The combined EtOAc extract was concentrated under reduced pressure and the residue was purified with e-phase prep-HPLC to afford 101 (55 mg, 40%). LCMS: [M+H]+ 637. 1H NMR (500 MHz, DMSO) δ 8.58 (s, 1H), 8.55 (s, 1H), 8.49 (s, 1H), 8.41 (s, 1H), 7.86 (d, J=3.0, 1H), 7.38-7.37 (m, 2H), 7.25-7.23 (m, 1H), 10 6.54 (s, 1H) , 5.16 (t, J=3.0, 1H), 4.56 -4.40 (m, 6H), 4.19 -4.12 (m, 3H), 3.95 (t, J=3.0, 1H), 3.60 (s, 3H), 3.43-3.41(m, 1H), 3.06 (s, 4H), 2.57-2.61 (m, 2H), 2.45-2.48 (m, 6H), 1.78-1.80 (m, 2H), 1.69-1.70 (m, 2H) Example 102a 1-Methyl[5-(4-methyl-piperazinyl)-pyridinylamino] (4,4,5,5-tetramethyl-[1,3,2]dioxaborolanyl)-1H-pyridinone 102a N N N NH O O N B O 15 102a A 1-L single-neck round-bottomed flask equipped with a magnetic stirrer and thermoregulator was purged with nitrogen and charged with 5-bromomethyl[5-(4- methylpiperazinyl)-pyridinylamino]-1H-pyridinone prepared according to US 2009/0318448, (10.0 g, 0.027 mol), bis(pinacolato)diboron (8.06 g, 0.032 mol), ium 20 acetate (10.4 g, 0.11 mol) and 1,4-dioxane (200 mL). After a stream of nitrogen was passed through the resulting suspension for 30 min., f)Cl2 CH2Cl2 (582 mg, 0.795 mmol) was added. The ing reaction mixture was stirred at reflux for 3 h. Then, it was cooled to room temperature, ioned between water (400 mL) and ethyl acetate (600 mL) and filtered through a pad of CELITE®. The organic phase was extracted, dried over sodium 25 sulfate, filtered and concentrated. The residue was ated with a mixture of diethyl ether (50 mL) and hexanes (250 mL), and the suspension was filtered. The filter cake was dried under vacuum at room temperature to afford a 27 % yield (3.04 g) of 1-methyl[5-(4- methyl-piperazinyl)-pyridinylamino](4,4,5,5-tetramethyl-[1,3,2]dioxaborolanyl)- 1H-pyridinone 102a as a brown solid. 104 Example 102b 3-(1-Methyl(5-(4-methylpiperazinyl)pyridinylamino)oxo- 1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- nicotinaldehyde 102b A sealed tube was charged with 3-bromo(1-oxo-3,4,6,7,8,9- 5 hexahydropyrazino[1,2-a]indol-2(1H)-yl)isonicotinaldehyde 101f (200 mg, 0.53 mmol), 1- methyl(5-(4-methylpiperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2- dioxaborolanyl)pyridine-2(1H)-one 102a (225 mg, 0.53 mmol), PdCl2(dppf) (42 mg, 0.05mmol), K3PO4 (210 mg, 1 mmol), and NaOAc (85mg, 1 mmol) in acetonitrile/H2O (8 mL/1 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 4 h. 10 It was then filtered and the filtrate was evaporated in vacuo. The e was purified on flash column eluting with 20:1 DCM/MeOH to afford 102b (135 mg, 43%). LCMS: [M+H]+ 593 . e 102 2-(4-(Hydroxymethyl)(1-methyl(5-(4-methylpiperazin yl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-1(2H)-one 102 15 A mixture of 3-(1-methyl(5-(4-methylpiperazinyl)pyridinylamino)oxo- 1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- yl)isonicotinaldehyde 102b (135 mg, 0.22 mmol) and NaBH4 (20 mg, 0.5 mmol) in MeOH (5 mL) was stirred at 0 oC for 0.5 h. The mixture was quenched with water and the residue was extracted with EtOAc (5 mL X 2). The combined EtOAc extract was concentrated under 20 reduced pressure and the residue was purified with reverse-phase prep-HPLC to afford 102 (18 mg, 20%). LCMS: [M+H]+ 595. 1H NMR (500 MHz, DMSO) δ 8.59 (s, 1H), 8.55 (s, 1H), 8.49 (s, 1H), 8.45 (s, 1H), 7.87 (s, 1H), 7.37-7.38 (m, 2H), 7.23-7.25 (m, 1H), 6.54 (s, 1H) , 5.16 (t, J=3.0, 1H), 4.40 (s, 2H), 4.14-4.18 (m, 3H), 3.93-3.95 (m, 1H), 3.60 (s, 3H), 3.09 (s, 4H), .61(m, 6H), 2.48-2.34 (m, 5H), 1.78-1.79 (m, 2H), 1.69-1.70 (m, 2H) 25 Example 103a 2-Bromochloronicotinaldehyde 103a To a solution of 2-bromochloropyridine (1.6 g, 8.0 mmol) in anhydrous ydrofuran (40 mL) cooled at -70oC was added the solution of lithium ropyl-amide (5.0 mL, 10.0 mmol, 2.0 M) over a period of 5 minutes and stirred at -70 oC for another 1 h.
Anhydrous DMF (1.3 g) was introduced over a period of 3 minutes and the mixture was 30 stirred for another 30 minutes. It was then quenched with saturated NH4Cl (30 mL) and extracted with ethyl acetate (20 mL × 3). The combined organic layer was dried over ous , filtered, and evaporated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with eum ether/ethyl acetate (20:1) to 105 afford 103a as a yellow solid (900 mg, 48%). 1H NMR (500 MHz, DMSO) δ 10.21 (s, 1H), 8.52 (d, J = 5.5 Hz, 1H) , 7.79 (d, J = 5.0 Hz, 1H).
Example 103b 4-Chloro(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl)nicotinaldehyde 103b 5 A 100-mL single-neck round-bottomed flask equipped with a ic stirrer and a reflux condenser was charged with 103a (800 mg, 3.5 mmol), 3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-1(2H)-one 101e (665 mg, 3.5 mmol), tris(dibenzylideneacetone)dipalladium(0) (320 mg, 0.35 mmol), XantPhos (400 mg, 0.7 mmol), Cs2CO3 (2.3 g, 7.0 mmol), and oxane (20 mL). After three cycles of 10 /argon flush, the mixture was heated at 90oC for 5 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (80:1) to afford 103b as a yellow solid (1.2 g, 50%). MS: [M+H]+ 330. 15 Example 103c 4-(1-Methyl(5-(4-(oxetanyl)piperazinyl)pyridin ylamino)oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)nicotinaldehyde 103c A 100-mL single-neck bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 103b (600 mg, 1.0 mmol), 1-methyl(5-(4-(oxetan 20 yl)piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxa-borolan yl)pyridin-2(1H)-one 101l (468 mg, 1.0 mmol), Pd(dppf)Cl2 (81 mg, 0.1 mmol), K3PO4.3H2O (678 mg, 3.0 mmol), and tetrahydrofuran (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at reflux for 4 h. It was then cooled to room temperature and filtered.
The filtrate was concentrated under reduced pressure and the resulting residue was purified 25 by silica-gel column chromatography eluting with dichloromethane/methanol (40:1) to afford 103c as yellow solid (510 mg, 73%). MS: [M+H]+ 635.
Example 103 2-(3-(Hydroxymethyl)(1-methyl(5-(4-(oxetanyl)piperazin yl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)-3,4,6,7,8,9- dropyrazino[1,2-a]indol-1(2H)-one 103 30 To the solution of 103c (500 mg, 0.8 mmol) in ol (50 mL) was added sodium borohydride (91 mg, 2.4 mmol) at 0 oC and stirred for r 30 minutes. Then the reaction mixture was quenched with water (3 mL) and concentrated. The residue was purified with e-phase prep-HPLC to afford 103 (224 mg, 45%). LCMS: [M+H]+ 637. 1H NMR (500 MHz, DMSO) δ 8.61 (d, J=3.0, 1H), 8.48 (d, J=6.0, 1H), 7.92 (d, J=3.5, 1H), 7.81(d, J=3.0, 106 1H), 7.78 (s, 1H), 7.38 (d, J=6.0, 1H), 7.24-7.27 (m, 1H), 6.88 (s, 1H), 6.81 (d, J=11.5,1H), 5.01-5.04, (m, 1H), 4.60-4.71 (m, 5H), 4.32-4.49 (m, 2H), 3.83-4.15 (m, 3H), 3.70(s, 3H), 3.53-3.59 (m, 1H), 3.13-3.16 (m, 4H), 2.55-2.61 (m, 4H), 2.49-2.52 (m, 4H), 1.78-1.90 (m, 4H) 5 Example 104a 4-Bromochloronicotinaldehyde 104a To a solution of 4-bromochloropyridine (12.0 g, 60.0 mmol) in anhydrous tetrahydrofuran (300 mL) cooled at -70 oC was added the solution of lithium diisopropylamide (30.0 mL, 60.0 mmol, 2.0 M) over a period of 30 minutes and stirred for 10 r at -70 oC 2 h. Anhydrous DMF (12.0 g) was introduced over a period of 10 minutes and stirred for another 30 minutes. It was then quenched with saturated NaHCO3 (200 mL), extracted with ethyl acetate (100 mL × 3). The combined organic layer was dried over anhydrous Mg2SO4, filtered, and evaporated under reduced pressure. The residue was ed by silica-gel column tography eluting with petroleum ether/ethyl acetate (20:1) to 15 afford 104a as a yellow solid (4.0 g, 29%). 1H NMR (500 MHz, DMSO) δ 10.23 (s, 1H), 8.44 (d, J = 5.5 Hz, 1H) , 7.94 (d, J = 5.5 Hz, 1H).
Example 104b 2-Chloro(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl)nicotinaldehyde 104b 20 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 104a (1.1 g, 5.0 mmol), 3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-1(2H)-one 101e (477 mg, 2.5 mmol), tris(dibenzylideneacetone)dipalladium(0) (230 mg, 0.25 mmol), os (430 mg, 0.75 mmol), Cs2CO3 (1.6 g, 5.0 mmol), and 1,4-dioxane (50 mL). After three cycles of 25 vacuum/argon flush, the mixture was heated at 65oC for 2 h. It was then cooled to room temperature and filtered. The te was concentrated under reduced pressure and the ing residue was purified by -gel column chromatography eluting with 107 dichloromethane/methanol (40:1) to afford 104b as a yellow solid (1.1 g, 80%). MS: [M+H]+ 330.
Example 104c 2-(1-Methyl(5-(4-(oxetanyl)piperazinyl)pyridin ylamino)oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- 5 a]indol-2(1H)-yl)nicotinaldehyde 104c A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 104b (658 mg, 1.0 mmol), 1-methyl(5-(4-(oxetan yl)piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxa-borolan yl)pyridin-2(1H)-one 101l (622 mg, 2.0 mmol), Pd (dppf) Cl2 (65 mg, 0.08 mmol), 10 K3PO4.3H2O (361 mg, 1.6 mmol), and tetrahydrofuran (40 mL). After three cycles of vacuum/argon flush, the e was heated at reflux for 4 h. It was then cooled to room temperature and ed. The te was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (40:1) to afford 104c as a yellow solid (400 mg, 63%). MS: 15 [M+H]+ 635.
Example 104 2-(3-(Hydroxymethyl)(1-methyl(5-(4-(oxetan yl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 104 To the solution of 104c (360 mg, 0.6 mmol) in methanol (50 mL) was added sodium 20 borohydride (70 mg, 1.8 mmol) at 0 oC and stirred for another 30 minutes. Then the reaction mixture was quenched with water (2 mL) and concentrated. The residue was purified with reverse-phase PLC to afford 104 (63 mg, 16%) as an off-white solid. LCMS: [M+H]+ 637. 1H NMR (500 MHz, DMSO) δ 8.70 (d, J=3.0, 1H), 8.65 (d, J=5.5, 1H), 8.34 (s, 1H), 7.85(d, J=3.0, 1H), 7.60 (d, J=2.5, 1H), 7.36-7.37 (m, 2H), 7.22-7.23 (m, 1H), 6.56 (s, 1H), 25 5.12 (t, J=5.5, 1H), 4.55-4.56 (m, 2H), 4.43-4.45 (m, 4H), 4.14-4.16 (m, 3H), 3.93-3.95 (m, 1H), 3.60 (s, 3H), 3.43-3.44 (m, 1H), 3.05-3.08 (m, 4H), 2.61-2.63 (m, 2H), 2.46-2.47 (m, 2H), 2.36-2.39 (m, 4H), 1.68-1.78 (m, 4H).
Example 105a N-Methoxy-N-methyl-4,5,6,7-tetrahydrobenzo[b]thiophene carboxamide 105a 30 108 A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer was purged with nitrogen, charged with 4,5,6,7-tetrahydrobenzo[b]thiophenecarboxylic acid (3.00 g, 16.5 mmol), methylene chloride (80 mL), and DMF (60 mg, 0.825 mmol) and cooled to 0 °C. To the resulting solution, oxalyl chloride (2.31 g, 18.2 mmol) was added dropwise. 5 After this addition was complete, the reaction was warmed to room temperature and stirred for 2 h. After this time, the reaction was concentrated to dryness under reduced re. The resulting white solid was ved in methylene chloride (80 mL) and the solution cooled to 0 °C. Triethylamine (5.00 g, 49.5 mmol) and methylhydroxylamine (1.61 g, 16.5 mmol) were then added. After the addition was complete, the g bath was removed, and 10 the reaction mixture was stirred at room temperature for 16 h. After this time, the reaction mixture was partitioned between water (100 mL) and ethyl acetate (200 mL). The layers were separated, and the aqueous phase was extracted with ethyl acetate (100 mL). The combined c extracts were washed with water (100 mL), followed by brine (100 mL) and dried over sodium sulfate. The drying agent was d by filtration, and the solvent was 15 evaporated under reduced pressure. The resulting residue was purified by flash chromatography to afford an 88% yield of 105a (3.29 gm) as a white solid: mp 36–37 °C; 1H NMR (500 MHz, CDCl3) δ 7.79 (s, 1H), 3.76 (s, 3H), 3.34 (s, 3H), 2.78 (t, 2H, J = 6.0 Hz), 2.62 (t, 2H, J = 6.0 Hz), 1.82 (m, 4H); MS (APCI+) m/z 226.3 (M+H) Example 105b 3-Chloro(4,5,6,7-tetrahydrobenzo[b]thiophenyl)propan 20 one 105b Cl S O 105b A 100-mL single-necked round-bottomed flask equipped with a magnetic stirrer was purged with nitrogen and charged with 105a (2.70 g, 12.0 mmol) and anhydrous THF (45 mL), and the solution was cooled to -10 °C with e/ice bath. A 1.0 M solution of 25 vinylmagnesium bromide in THF (13.2 mL, 13.2 mmol) was added dropwise, and the resulting reaction mixture was d at 0 °C for 4 h. After this time, the reaction mixture was partitioned between ethyl acetate (100 mL) and 2 M s hydrochloric acid (40 mL).
The layers were separated, and the aqueous phase was extracted with ethyl acetate (40 mL).
The combined organic extracts were washed with water (100 mL), followed by brine (100 30 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The 109 resulting residue was dissolved in ene chloride (30 mL), and a 2 M solution of hydrogen chloride in diethyl ether (15 mL) was added. After stirring at room temperature for 1 h, the solvents were removed under reduced pressure. Purification of the resulting residue by column chromatography afforded a 29% yield (804 mg) of 105b as an off-white solid: mp 5 57–58 °C; 1H NMR (500 MHz, CDCl3) δ 7.41 (s, 1H), 3.89 (t, 2H, J = 7.0 Hz), 3.30 (t, 2H, J = 7.0 Hz), 2.81 (t, 2H, J = 6.0 Hz), 2.64 (t, 2H, J = 6.0 Hz), 1.83 (m, 4H); MS (ECI+) m/z 229.1 (M+H) Example 105c 5,6,7,8-Tetrahydro-1H-benzo[b]cyclopenta[d]thiophen-3(2H)- one 105c 10 A 50-mL single-necked round-bottomed flask equipped with a magnetic stirrer was charged with 105b (800 mg, 3.51 mmol) and 98% sulfuric acid (8 mL). After stirring at 95 °C for 16 h, the reaction e was poured into ice (50 g), and the ing suspension was ted with ethyl e (3 × 50 mL). The organic extracts were combined, dried over 15 sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography to afford 105c in 47% yield (320 mg) as an off-white solid: mp 75–76 °C; 1H NMR (500 MHz, CDCl3) δ 2.89 (m, 2H), .83 (m, 4H), 2.56 (t, 2H, J = 6.5 Hz), 1.84 (m, 4H) Example 105d 5,6,7,8-Tetrahydro-1H-benzo[b]cyclopenta[d]thiophen-3(2H)- 20 one oxime 105d A 100-mL single-neck round-bottomed flask equipped with a mechanical stirrer and nitrogen inlet was charged with hydroxylamine hydrochloride (573 mg, 8.25 mmol) and methanol (10 mL). The mixture was cooled to 0 ºC using an ice bath. Sodium acetate (677 25 mg, 8.25 mmol) was added. The mixture was stirred at 0 ºC for 30 min. After this time, 105c (319 mg, 1.65 mmol) was added, and the reaction was stirred at room temperature for 16 h.
After this time, the mixture was concentrated, and the resulting residue was triturated with water (10 mL). The resulting solid was collected and dried in a vacuum oven at 45 °C to afford an 84% yield (287 mg) of 105d as an off-white solid: mp 173–174 °C; 1H NMR (500 110 MHz, DMSO-d6) δ 10.38 (s, 1H), 2.97 (m, 2H), 2.77–2.73 (m, 4H), 2.47 (m, 2H), 1.75 (m, 4H); MS (APCI+) m/z 208.3 (M+H) Example 105e 3,4,5,6,7,8-Hexahydrobenzothieno[2,3-c]pyridin-1(2H)-one 105e 5 A 50-mL single-neck round-bottomed flask equipped with a reflux condenser, ic stirrer and nitrogen inlet was charged with 105d (285 mg, 1.38 mmol) and polyphosphoric acid (15 g). After stirring at 80 ºC for 16 h, the reaction mixture was cooled to room temperature, and water (30 mL) was added. The resulting mixture was stirred for 30 10 min and filtered. The filter cake was washed with water (20 mL) and dried in a vacuum oven at 45 °C to afford a 75% yield (215 mg) of 105e as an off-white solid: mp 203 °C dec; 1H NMR (500 MHz, CDCl3) δ 5.62 (s, 1H), 3.59 (t, 2H, J = 7.0 Hz), 2.81 (t, 2H, J = 6.0 Hz), 2.72 (t, 2H, J = 7.0 Hz), 2.48 (t, 2H, J = 6.0 Hz), 1.84 (m, 4H). MS (APCI+) m/z 208.3 (M+H) 15 Example 105f 3-Bromo{6-oxothiaazatricyclo[7.4.0.02,7]trideca- 1(9),2(7)-dienyl}pyridinecarbaldehyde 105f To a 100-mL single-neck round-bottomed flask equipped with a magnetic r and reflux ser was charged with oxane (15 mL), 3,5-dibromoisonicotin-aldehyde 20 (400mg, 1.5 mmol), 8-thiaazatricyclo[7.4.0.02,7]trideca-1(9),2(7)-dienone 105e (146 mg, 0.76 mmol), and cesium carbonate (176 mg, 1.5 mmol). Xantphos (40 mg, 0.08 mmol) and Pd2(dba)3 (70 mg, 0.08 mmol) were added, and the reaction mixture was heated at 100 ºC for 5 h. After this time the reaction was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the residue was purified on flash column eluting 25 with DCM:MeOH (20:1) to afford 105f (200 mg, 70%). MS: [M+H]+ 377. 111 Example 105g 3-[1-Methyl({5-[4-(oxetanyl)piperazinyl]pyridine yl}amino)oxo-1,6-dihydropyridinyl]{6-oxothiaazatricyclo[7.4.0.02,7]trideca- 1(9),2(7)-dienyl}pyridinecarbaldehyde 105g O N N S N NH O O O N N N 5 A sealed tube was charged with 105f (200 mg, 0.53 mmol), 1-methyl(5-(4-(oxetan- 3-yl) piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolan yl)pyridin-2(1H)-one 101l (240 mg, 0.51 mmol), PdCl2(dppf) (42 mg, 0.05 mmol), K3PO4 (230 mg, 1 mmol), and NaOAc (80 mg, 1 mmol) in CH3CN (5 mL) and H2O (1.5 mL). The system was ted and refilled with N2. The reaction mixture was heated at 100 ºC for 2 h. 10 It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was ed by flash column tography eluting with 10:1 of DCM/MeOH to afford 105g in 40% yield (138 mg) as a pale yellow solid. MS: [M+H]+ 638.
Example 105 4-Hydroxymethyl- 3-[1-methyl({5-[4-(oxetanyl)piperazin 15 yl]pyridineyl}amino)oxo-1,6-dihydropyridinyl]{6-oxothiaazatricyclo- [7.4.0.02,7]trideca-1(9),2(7)-dienyl}pyridine 105 To a solution of 3-[1-methyl({5-[4-(oxetanyl)piperazinyl]pyridine yl}amino)oxo-1,6-dihydropyridinyl]{6-oxothiaazatricyclo[7.4.0.02,7]-trideca- 1(9),2(7)-dienyl}pyridinecarbaldehyde 105g (130 mg, 0.20 mmol) in ol (5 mL) 20 at 0oC was added sodium borohydride (22 mg, 0.6 mmol) and stirred for 30 s. Then the reaction mixture was quenched with water (1.0 mL) and concentrated. The residue was purified by reverse-phase prep-HPLC to afford 105 (90 mg, 65 %). LCMS: [M+H]+ :654. 1H NMR (500 MHz, DMSO) δ 8.58 (d, J=2.0, 1H), 8.56 (s, 1H), 8.49 (s, 1H), 8.41 (s, 1H), 7.86 (s, 1H), 7.36 (m, 2H), 7.24-7.22 (m,1H), 5.14 (t, J=3.0, 1H), 4.56-4.42 (m, 6H), 4.08-3.90 (m, 25 2H), 3.60 (s, 3H), 3.43 (d, J=3.5,1H), 3.07 (s, 4H), 2.89-2.79 (m, 4H) , 2.55-2.53 (m, 2H), 2.39-2.37 (m, 4H), 1.80-1.81 (m, 4H) Example 106a 3,3-Dimethylcyclopentanone 106a 112 A 1-L three-neck round-bottomed flask ed with a magnetic stirrer, addition funnel and nitrogen inlet was purged with en and charged with ether (200 mL) and copper (I) iodide (54.46 g, 0.286 mol). The mixture was cooled to 0 °C, lithium (1.6 5 M in ether, 357.5 mL, 0.572 mol) was added dropwise to the reaction mixture over 1.5 h and stirred at 0 oC for onal 2 h. After this time a solution of 3-methylcyclo-pentenone (25 g, 0.260 mol) in ether (150 mL) was added dropwise over 1.5 h. The reaction mixture was then stirred at 0 oC for 2 h and poured into sodium sulfate deca-hydrate (300 g). The resulting mixture was stirred for 30 min. After this time the mixture was filtered and washed 10 with ether (1000 mL). The filtrate was concentrated and distilled under d pressure to afford a 70% yield (20.5 g) of 3,3-dimethylcyclo-pentanone 106a as a colorless liquid: bp 50–55 °C (at 10 mmHg); 1H NMR (300 MHz, CDCl3) δ 2.31 (t, 2H, J = 7.8 Hz), 2.05 (s, 2H), 1.79 (t, 2H, J = 7.8 Hz); MS (ESI+) m/z 113.3 (M+H) Example 106b Ethyl 5,5-Dimethyl-5,6-dihydro-4H-cyclopenta[b]thiophene 15 carboxylate 106b A 500-mL three-neck round-bottomed flask equipped with a magnetic stirrer, reflux condenser, addition funnel and nitrogen inlet was purged with nitrogen and charged with DMF (9.49 g, 0.100 mol) and methylene chloride (100 mL). The on mixture was 20 cooled to 0 °C and phosphorus oxychloride (14.1 g, 0.920 mol) was added dropwise to the reaction over 30 min. Once this addition was complete, the reaction was warmed to room temperature and stirred for 1 h. After this time a solution of 106a (11.2 g, 0.100 mol) in methylene de (100 mL) was added dropwise over 1 h. The reaction was then stirred at reflux for 18 h. The reaction mixture was cooled to room temperature and poured into a 25 mixture of crushed ice (400 mL) and sodium acetate (100 g, 1.22 mol). The resulting mixture was stirred for 45 min. After this time the aqueous layer was separated and extracted with methylene chloride (2 × 500 mL). The combined organic layers were then washed with water (2 × 200 mL), followed by brine (200 mL) and dried over sodium e. The drying agent was then removed by filtration, and the filtrate was concentrated to afford crude product 2- 30 chloro-4,4-dimethylcyclopentenecarbaldehyde which was placed in a 500-mL three-neck 113 round bottomed flask equipped with a mechanical r, reflux ser and nitrogen inlet.
Methylene chloride (200 mL), ethyl 2-mercaptoacetate (11.0 g, 0.092 mol) and triethylamine (30 g, 0.207 mol) were then added. The on mixture was then stirred at reflux for 6 h.
After this time the reaction was cooled to room temperature and concentrated to a thick 5 orange residue. Ethanol (200 mL) and triethylamine (30.0 g, 0.207 mol) were added and the reaction was heated at reflux for 12 h. The reaction was then cooled to room temperature and concentrated under reduced pressure and the ing residue was diluted with ether (600 mL). The resulting mixture was washed with 1 M hydrochloric acid (150 mL), brine (100 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The 10 resulting residue was purified by flash chromatography to afford 106b in 34% yield (7.70 g) as a colorless liquid: 1H NMR (300 MHz, CDCl3) δ 7.48 (s, 1H), 4.33 (q, 2H, J = 7.2 Hz), 2.72 (s, 2H), 2.56 (s, 2H), 1.38 (t, 3H, J = 1.8 Hz), 1.17 (s, 6H); MS (ESI+) m/z 225.1 Example 106c 5,5-Dimethyl-5,6-dihydro-4H-cyclopenta[b]thiophene carboxylic acid 106c 15 In a 250-mL -neck round-bottomed flask equipped with a magnetic stirrer and reflux condenser, 106b (4.00 g, 17.8 mmol) was dissolved in ethanol (50 mL). THF (50 mL), water (50 mL) and lithium hydroxide (854 mg, 35.6 mmol) were added, and the mixture was stirred at 60 °C for 4 h. After this time the reaction was cooled to room temperature and 20 ied with 2M hydrochloric acid to pH 1.5, and then extracted with ethyl acetate (2 × 200 mL). The organic layers were combined, washed with water (2 × 100 mL), followed by brine (100 ml) and dried over sodium sulfate. The drying agent was then separated by filtration.
After ating the resulting filtrate, 106c was obtained in 91% yield (3.2 g) as a white solid: mp 170–172 °C; 1H NMR (300 MHz, CDCl3) δ 12.77 (s, 1H), 7.46 (s, 1H), 2.71 (s, 2H), 25 2.53 (s, 2H), 1.20 (s, 6H); MS (ESI–) m/z 195.0 Example 106d 5,5-Dimethyl-5,6-dihydro-4H-cyclopenta[b]thiophene carboxylic acid 106d A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer, reflux 30 condenser and a bubbler placed on the condenser was charged with 106c (2.30 g, 11.6 mmol), 114 toluene (25 mL), thionyl chloride (4.09 g, 34.9 mmol) and DMF (1 drop). The mixture was heated at reflux for 1 h and then evaporated under reduced pressure on a rotary evaporator at 45 °C. The ing acid chloride was d with methylene chloride (20 mL).
In a separate 250-mL three-neck round-bottomed flask equipped with a magnetic 5 stirrer N,O-dimethylhydroxylamine hydrochloride (2.26 g, 23.2 mmol) and N,N- diisopropylethylamine (2.97 g, 23.0 mmol) were dissolved in ous methylene chloride (20 mL) under nitrogen, and the solution was cooled to 0 °C in an ice/water bath. The solution of the acid chloride was added, and the reaction mixture was stirred at room temperature for 18 h. The reaction mixture was extracted with water (100 mL), 10% aqueous 10 citric acid (50 mL) and a 1:1 mixture of saturated aqueous sodium bicarbonate and water (100 mL). The organic layer was dried over sodium sulfate and evaporated under reduced pressure on a rotary evaporator to afford a 93% yield (2.60 g) of 106d as a light yellow solid: 1H NMR (300 MHz, CDCl 3) δ 7.66 (s, 1H), 3.77 (s, 3H), 3.35 (s, 3H), 2.74 (s, 2H), 2.58 (s, 2H), 1.23 (s, 6H) 15 Example 106e 3-Chloro(5,5-dimethyl-5,6-dihydro-4H- cyclopenta[b]thiophenyl)propanone 106e A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer was purged with nitrogen and charged with 106d (2.41 g, 10.0 mmol) and anhydrous THF (20 20 mL). The solution was cooled to -70 °C, and 1 M vinylmagnesium bromide in THF (11 mL, 11.0 mmol) was added with the reaction ature maintained below -60 °C. The reaction mixture was stirred at -13 to -7 °C for 2 h and then warmed to room temperature over 30 min.
The reaction was again cooled to -70 °C, and a 2 M solution of en chloride in ether (22.5 ml, 45 mmol) was added. The reaction was then stored in a freezer at -10 °C overnight. 25 After this time the e was evaporated under reduced pressure on a rotary evaporator, and the resulting residue partitioned between water (100 mL) and ether (100 mL). The ether extract was dried over sodium sulfate and evaporated under reduced re on a rotary ator to afford crude 106e (2.86 g, 118%) as a brown oil with approximately 75% purity (by NMR): 1H NMR (300 MHz, CDCl3) δ 7.45 (s, 1H), 3.89 (t, 2H, J = 6.9 Hz), 3.30 (t, 2H, J 30 =6.9 Hz), 2.75 (s, 2H), 2.59 (s, 2H), 1.24 (s, 6H) 115 Example 106f 6,6-Dimethyl-1,2,6,7-tetrahydrodicyclopenta[b,d]thiophen- 3(5H)-one 106f A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer was 5 d with crude 106e (2.86 g, 10.0 mmol presuming quantitative yield) and 98% ic acid. The reaction e was heated in a 90 °C oil bath overnight. The reaction e was placed into an ice/acetone bath, and a cold (5 °C) solution of dipotassium hydrogen phosphate (105 g, 0.603 mol) in water (300 mL) was added in one portion. The resulting mixture was shaken with ethyl acetate (300 mL) and filtered. The filter cake was washed 10 with ethyl acetate (100 mL). The ethyl acetate layer of the filtrate was separated, dried over sodium sulfate and evaporated under reduced pressure on a rotary evaporator. the resulting residue was purified by flash column chromatography (silica, 80:20 s/ethyl acetate) to afford 106f in 37% yield over two steps (683 mg) as an amorphous brown solid: mp 60– 62 °C; 1H NMR (500 MHz, CDCl3) δ 2.92–2.87 (m, 4H), 2.79 (s, 2H), 2.53 (s, 2H), 1.26 (s, 15 6H); LCMS (ESI+) m/z 207.0 (M+H) Example 106g 6,6-Dimethyl-1,2,6,7-tetrahydrodicyclopenta[b,d]thiophen- 3(5H)-one 106g A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and 20 nitrogen inlet was charged with hydroxylamine hloride (688 mg, 9.90 mmol), sodium acetate (812 mg, 9.90 mmol) and ol (10 mL), and the mixture at room temperature for 30 min. After this time, a solution of 106f (680 mg, 3.30 mmol) was added dropwise at room temperature, and the reaction was stirred at room temperature for 14 h under nitrogen here. Since the reaction was not complete, hydroxylamine hydrochloride (1.15 g, 16.5 25 mmol) and sodium acetate (1.35 g, 16.5 mmol) were added, and the stirring was continued at room temperature for 58 h. After this time, the mixture was diluted with methylene chloride (150 mL) and water (100 mL), and the layers were separated. The organic layer was washed with brine (50 mL) and dried over sodium sulfate. The drying agent was removed by filtration and the filtrate was concentrated to afford crude 106g in quantitative yield (730 mg) 116 as a yellow semi-solid which was used in the next step without purification: mp 122–124 ºC; 1H NMR for major isomer (500 MHz, CDCl 3) δ 3.13–3.11 (m, 2H), .83 (m, 2H), 2.77 (s, 2H), 2.49 (s, 2H), 1.24 (s, 6H); MS (ESI+) m/z 222.0 (M+H) Example 106h 6,6-Dimethyl-3,4,6,7-tetrahydro-5H-cyclopenta[4,5]thieno[2,3- 5 c]pyridine-1(2H)-one 106h A 100-mL three-neck round-bottomed flask equipped with a reflux ser, mechanical stirrer and nitrogen inlet was charged with 106g (700 mg, 3.16 mmol) and polyphosphoric acid (25 g). The reaction mixture was stirred at 80 ºC for 13 h under en 10 atmosphere. After this time, the e was cooled to 0 ºC and water (50 mL) was added dropwise carefully maintaining the internal temperature between 10–45 ºC. The mixture was diluted with 90:10 methylene chloride/methanol (100 mL) and the layers were ted.
The aqueous layer was ted with 90:10 methylene chloride/methanol (50 mL), and the combined organic layers were washed with saturated aqueous sodium bicarbonate (50 mL), 15 brine (150 mL) and dried over sodium sulfate. The drying agent was removed by filtration.
The filtrate was concentrated under reduced pressure, and the resulting residue was purified by flash column chromatography (silica, 95:5 methylene chloride/methanol) to afford 6,6- dimethyl-3,4,6,7-tetrahydro-5H-cyclopenta[4,5]thieno[2,3-c]pyridine-1(2H)-one 106h in 90% yield (630 mg) as an amorphous off-white solid: mp 205–207 °C; 1H NMR (500 MHz, 20 CDCl3) δ 5.51 (s, 1H), 3.60–3.56 (m, 2H), 2.76–2.73 (m, 4H), 2.49 (s, 2H), 1.26 (s, 6H); MS (ESI+) m/z 222.0 (M+H) Example 106i o{4,4-dimethyloxothia azatricyclo[6.4.0.02,6]dodeca-1(8), 2(6)-dienyl}pyridinecarbaldehyde 106i 25 To a 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and reflux condenser was charged with 1,4-dioxane (15 mL), 3,5-dibromoisonicotin-aldehyde (400mg, 1.5 mmol), 4,4-dimethylthiaazatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dien 117 one (106h) (170 mg, 0.76 mmol), and cesium carbonate (176 mg, 1.5 mmol). os (40 mg, 0.08 mmol) and Pd2(dba)3 (70 mg, 0.08 mmol) were added, and the on mixture was heated at 100 ºC for 5 h. After this time the reaction was cooled to room temperature and filtered. The filtrate was concentrated under d pressure and the residue was purified on 5 flash column eluting with DCM:MeOH (20:1) to afford 106i (200 mg, 65%). MS: [M+H]+ 405.
Example 106j 3-[1-Methyl({5-[4-(oxetanyl)piperazinyl]pyridine yl}amino)oxo-1,6-dihydropyridinyl]{4,4-dimethyloxothia azatricyclo[6.4.0.02,6]dodeca-1(8), 2(6)-dienyl}pyridinecarbaldehyde 106j O N N N NH O O S N N O N 10 106j A sealed tube was charged with 106i (200 mg, 0.50 mmol), yl(5-(4-(oxetan- 3-yl) piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolan yl)pyridin-2(1H)-one 101l (240 mg, 0.51 mmol), PdCl2(dppf) (42 mg, 0.05 mmol), K3PO4 (230 mg, 1 mmol), and NaOAc (80 mg, 1 mmol) in CH3CN (5 mL) and H2O (1.5 mL). The 15 system was evacuated and refilled with N2. The reaction mixture was heated at 100 ºC for 2 h.
It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by flash column chromatography g with 10:1 DCM/MeOH to afford 106j (130 mg, 40%) as a pale yellow solid. MS: [M+H]+ 666. 20 Example 106 4-Hydroxymethyl[1-methyl({5-[4-(oxetanyl)piperazin yl]pyridineyl}amino)oxo-1,6-dihydropyridinyl]{4,4-dimethyloxothia azatricyclo[6.4.0.02,6]dodeca-1(8), 2(6)-dienyl}pyridinecarbaldehyde 106 To a on of 106j (130 mg, 0.20 mmol) in methanol (5 mL) at 0 oC was added sodium borohydride (22 mg, 0.6 mmol) and stirred for 30 minutes. Then the reaction mixture 25 was quenched with water (1.0 mL) and concentrated. The residue was purified by reversephase prep-HPLC to afford 106 (60 mg, 45 %) as a yellow solid. LCMS: [M+H]+: 668. 1H 118 NMR (500 MHz, DMSO) δ 8.58 (d, J=2.0, 1H), 8.56 (s, 1H), 8.49 (s, 1H), 8.41 (s, 1H), 7.87 (d, J=2.5, 1H), .36 (m, 2H), 7.24-7.22 (m, 1H), 5.15 (t, J=5.0, 1H), 4.56-4.42 (m, 6H), 4.08-4.04 (m, 2H), 3.60 (s, 3H), 3.43-3.42 (m, 1H), 3.07-2.94 (m, 6H), 2.55-2.53 (m, 4H), 2.39-2.38 (m, 4H), 1.23 (s, 6H) 5 Example 107a (E)-Ethyl 3-(2-Chloro-4,4-dimethylcyclopentenyl)acrylate 107a Cl EtO2C 107a The following two procedures were adapted from Organic Preparations and ures Int., 29(4):471-498. A 500-mL single neck round bottomed flask equipped with 10 a magnetic stirrer and nitrogen inlet was charged with 2-chloro-4,4-dimethylcyclopent enecarbaldehyde (38 g, 240 mmol) in benzene (240 mL). To the solution was added ethoxycarbonylmethylene nylphosphorane (84 g, 240 mmol). The mixture was stirred for 14 h. After that time, the solvent was evaporated and the residue was triturated with hexanes (2 L) to extract the t away from the PPh3 by-products. The organic layer was 15 dried over sodium sulfate and concentrated in vacuo. The residue was purified by column chromatography using a 100% hexane – 1:1 /ethyl acetate gradient to afford a 37% yield (20 g) of (E)-ethyl 3-(2-chloro-4,4-dimethylcyclopentenyl)acrylate 107a.
Example 107b Ethyl 5,5-Dimethyl-1,4,5,6-tetrahydrocyclopenta[b]pyrrole carboxylate 107b 20 A 250-mL single neck round bottomed flask equipped with a magnetic r and nitrogen inlet was charged with 107a (17 g, 74 mmol) in DMSO (100 mL). To the solution was added sodium azide (9.6 g, 150 mmol). The mixture was then heated to 75 °C and stirred for 8 h. After cooling to rt (room temperature), H2O (100 mL) and CH2Cl2 (200 mL) 25 were added and the organic layer was separated. The aqueous layer was extracted with CH2Cl2 (50 mL). The combined organic layers were washed with brine, dried over sodium e and concentrated in vacuo. The residue was purified by column chromatography 119 using a 100% hexane – 1:1 hexane/ethyl acetate gradient to afford a 37% yield (5.7 g) of 107b.
Example 107c Ethyl 1-(Cyanomethyl)-5,5-dimethyl-1,4,5,6-tetrahydrocyclopenta rolecarboxylate 107c 5 A 250-mL single neck round bottomed flask equipped with a magnetic stirrer and nitrogen inlet was d with 107b (6.2 g, 30 mmol) in DMF (57 mL). To the solution was added NaH (80% dispersion in mineral oil, 1.26 g, 42.1 mmol). The resulting mixture was stirred at rt for 90 min. After that time, bromoacetonitrile (2.94 mL, 42 mmol) was added. 10 The mixture was stirred for 14 h. After that time, water (100 mL) and ethyl acetate (200 mL) were added and the organic layer was separated. The aqueous layer was extracted with ethyl e (2 X 50 mL). The combined organic layers were washed with brine, dried over sodium sulfate and concentrated in vacuo. The residue was purified by column chromatography to afford a 95% yield (7 g) of 107c. 15 Example 107d Ethyl 1-(2-Aminoethyl)-5,5-dimethyl-1,4,5,6-tetrahydrocyclopenta [b]pyrrolecarboxylate hloride 107d A 500-mL Parr reactor bottle was purged with nitrogen and d with 10% palladium on carbon (50% wet, 2.0 g dry weight), 107c (4.5 g, 18 mmol), 12% hydrochloric 20 acid (9.2 mL, 37 mmol), ethyl acetate (80 mL) and ethanol (52 mL). The bottle was attached to a Parr hydrogenator, evacuated, charged with hydrogen gas to a pressure of 50 psi and shaken for 6 h. After this time, the hydrogen was evacuated, and nitrogen was charged into the . CELITE® 521 (10.0 g) was added, and the mixture was filtered through a pad of CELITE® 521. The filter cake was washed with ethanol (2 × 50 mL), and the combined 25 filtrates were trated to dryness under reduced pressure. The crude residue ethyl 1-(2- 120 aminoethyl)-5,5-dimethyl-1,4,5,6-tetrahydrocyclopenta[b]pyrrolecarboxylate hydrochloride 107d was carried onto the next step without further purification.
Example 107e 4,4-Dimethyl-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dien- 9-one 107e 5 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and nitrogen inlet was purged with nitrogen and charged with crude 1-(2-aminoethyl)-5,5- dimethyl-1,4,5,6-tetrahydrocyclopenta[b]pyrrolecarboxylate hydrochloride 107d (~18 mmol), sodium de (6.2 g, 92 mmol) and ethanol (120 mL). The mixture was stirred at 10 55 oC over night. After that time, the reaction mixture was concentrated under reduced pressure and the residue was ioned between ethyl acetate (200 mL) and water (100 mL).
The solution was ed. The solid was washed with ethyl acetate (15 mL) to give 850 mg of desired product 107e. The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (2 × 100 mL). The combined organic layers were dried over 15 sodium sulfate and concentrated under reduced pressure to near dryness. The on was filtered and the solid (1.44 g) was washed with ethyl e (15 mL). The combined solids were dried under vacuum a afford 61% yield (2.3 g) of 107e.
Example 107f 3-Bromo{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 107f 20 A 100-mL single-neck bottomed flask equipped with a magnetic stirrer and reflux condenser was charged with 1,4-dioxane (15 mL), 3,5-dibromoisonicotinaldehyde (400 mg, 1.5 mmol), 107e (155 mg, 0.76 mmol), and cesium carbonate (176 mg, 1.5 mmol).
Xantphos (40 mg, 0.08 mmol) and Pd2(dba)3 (70 mg, 0.08 mmol) were added, and the reaction mixture was heated at 100 ºC for 5 h. After this time the reaction was cooled to 25 room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue was purified on flash column eluting with DCM:MeOH (20:1) to afford 107f (200 mg, 70%). MS: [M+H]+ 388.
Example 107g 5-[1-Methyl({5-[4-(oxetanyl)piperazinyl]pyridine yl}amino)oxo-1,6-dihydropyridinyl]{4,4-dimethyloxo-1,10- 30 diazatricyclo[6.4.0.02,6]-dodeca-2(6),7-dienyl}pyridinecarbaldehyde 107g 121 A sealed tube was charged with 107f (200 mg, 0.51 mmol), 1-methyl(5-(4-(oxetan- 3-yl)piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolan yl)pyridin-2(1H)-one 101l (240 mg, 0.51 mmol), PdCl2(dppf) (42 mg, 0.05 mmol), K3PO4 (230 mg, 1 mmol), and NaOAc (80 mg, 1 mmol) in CH3CN (5 mL) and H2O (1.5 mL). The 5 system was evacuated and refilled with N2.The reaction mixture was heated at 100 ºC for 2 h.
It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by flash column chromatography g with 10:1 of DCM/MeOH to afford 107g in 35% yield (120 mg) as a brown solid.
MS: [M+H]+ 649. 10 Example 107 [1-Methyl({5-[4-(oxetanyl)piperazinyl]pyridine yl}amino)oxo-1,6-dihydropyridinyl](hydroxymethyl)pyridinyl]-4,4-dimethyl- 1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienone 107 To a solution of 107g (120 mg, 0.18 mmol) in methanol (5 mL) at 0 oC was added sodium borohydride (22 mg, 0.6 mmol) and the mixture was stirred for 30 minutes. Then the 15 reaction mixture was quenched with water (1.0 mL) and concentrated. The residue was purified by reverse-phase prep-HPLC to afford 107 (72 mg, 60 %). LCMS: [M+H]+ :651. 1H NMR (500 MHz, CDCl3) δ 8.65 (s,1H), 8.59 (s, 1H), 8.49 (s, 1H), 7.86 (d, J=1.5, 1H), 7.36 (m, 2H), 7.22 (d, J=2.4, 2H), 6.52 (s, 1H), 5.16 (t, J=3.0, 1H), 4.56- 4.44 (m, 6H), .12 (m, 3H), 3.92 (m, 1H), 3.60 (s, 3H) , .42 (m, 1H), 3.06 (s, 4H), 2.57-2.38 (m, 8H) ,1.21 20 (s, 6H) Example 108a ro{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 108a A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 2-bromochloronicotinaldehyde 103a (3.0 g, 13.6 25 mmol), 4,4-dimethyl-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienone 107e (1.84 g, 9.0 mmol), tris(dibenzylideneacetone)dipalladium(0) (826 mg, 0.9 mmol), XantPhos (1.04 mg, 1.8 mmol), Cs2CO3 (5.8 g, 18.0 mmol), and 1,4-dioxane (40 mL). After three cycles of vacuum/argon flush, the mixture was heated at 90 oC for 5 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced re and the 30 resulting residue was tallized from ethyl acetate to afford 108a as a yellow solid (730 mg, purity: 99%; yield: 31.7 %). MS: [M+H]+ 344.0.
Example 108b 4-(1-Methyl(5-(4-(oxetanyl)piperazinyl)pyridin ylamino)oxo-1,6-dihydropyridinyl){4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]-dodeca-2(6),7-dienyl}nicotinaldehyde 108b 122 A 100-mL single-neck bottomed flask equipped with a ic stirrer and a reflux condenser was charged with ro{4,4-dimethyloxo-1,10- ricyclo [6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 108a (350 mg, 1.02 mmol), 1-methyl(5-(4-(oxetanyl)piperazinyl)pyridinylamino)(4,4,5,5- 5 tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 101l (476 mg, 1.02 mmol), Pd(dppf)Cl2 (83 mg, 0.10 mmol), K3PO4 (526 mg, 3.06 mmol), and ydrofuran (20 mL).
After three cycles of vacuum/argon flush, the mixture was heated at reflux for 4 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 10 dichloromethane/methanol (40:1) to afford 108b as white solid (400 mg, 61%). MS: [M+H]+ 649.4. e 108 2-(3-(Hydroxymethyl)(1-methyl(5-(4-(oxetanyl)piperazin yl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 4,4-dimethyl-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienone 108 15 To a solution of 4-(1-methyl(5-(4-(oxetanyl)piperazinyl)pyridinylamino)- 6-oxo-1,6-dihydropyridinyl){4,4-dimethyloxo-1,10-diazatricyclo-[6.4.0.02,6]dodeca- 2(6),7-dienyl}nicotinaldehyde 108b (400 mg, 0.62 mmol) in methanol (30 mL) at 0oC was added sodium borohydride (70 mg, 1.86 mmol) and stirred for 30 minutes. Then the reaction mixture was quenched with water (1.0 mL) and trated. The residue was 20 purified by reverse-phase prep-HPLC to afford 108 (170 mg, 42 %). LCMS: [M+H]+ 651.4. 1H NMR (500 MHz, CDCl 3) δ 8.63 (d, J=2.0, 1H), 8.48 (d, J= 5.0, 1H), 7.92 (d, J= 2.5, 1H), 7.82 (d, J = 2.5, 1H), 7.78 (s, 1H), 7.36 (d, J = 5.0,1H), 7.27-7.25 (m, 1H), 6.84 (s, 1H), 6.81 (d, J = 9.5, 1H), 5.05 (t, J = 6.5, 1H), 4.72-4.64 (m, 5H), 4.51-4.48 (m, 1H), 4.34-4.32 (m,1H), 4.15 (d, J = 4.5, 2H), 3.87-3.84 (m, 1H), 3.71 (s, 3H), 3.59-3.54 (m, 1H), 3.16-3.14 (m, 4H), 25 2.58-2.50 (m, 8H), 1.27(s, 6H) Example 109a 4-Chloro{4,4-dimethyloxothia azatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienyl}pyridinecarbaldehyde 109a A 100-mL single-neck round-bottomed flask equipped with a magnetic r and a reflux condenser was charged with 2-bromochloronicotinaldehyde 103a (660 mg, 3.0 30 mmol), 4,4-dimethylthiaazatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienone 106h (665 mg, 3.0 mmol), tris(dibenzylideneacetone)dipalladium(0) (270 mg, 0.3 mmol), XantPhos (340 mg, 0.6 mmol), Cs2CO3 (2.0 g, 6.0 mmol), and 1,4-dioxane (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at 90oC for 5 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the 123 ing residue was purified by -gel column chromatography eluting with dichloromethane to afford 109a as yellow solid (105 mg, 14%). MS: [M+H]+ 361.
Example 109b 4-(1-Methyl(5-(4-(oxetanyl)piperazinyl)pyridin ylamino)oxo-1,6-dihydropyridinyl)- 2-{4,4-dimethyloxothia 5 azatricyclo[6.4.0.02,6]-dodeca-1(8),2(6)-dienyl}nicotinaldehyde 109b A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with 109a (75 mg, 0.2 mmol), 1-methyl(5-(4-(oxetan yl)piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin- 2(1H)-one 101l (94 mg, 0.2 mmol), Pd(dppf)Cl2 (17 mg, 0.02 mmol), K3PO4.3H2O (140 mg, 10 0.6 mmol), and tetrahydrofuran (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at reflux for 4 h. It was then cooled to room ature and filtered. The filtrate was trated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (40:1) to 109b as yellow solid (60 mg, 47%). MS: [M+H]+ 666. 15 e 109 Hydroxymethyl)(1-methyl(5-(4-(oxetan yl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 4,4- dimethylthiaazatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienone 109 To a solution of 109b (60 mg, 0.1 mmol) in methanol (5 mL) at 0oC was added sodium dride (11 mg, 0.3 mmol) and the mixture was stirred for 30 minutes. Then the 20 reaction mixture was quenched with water (0.3 mL) and concentrated. The residue was purified with reverse-phase prep-HPLC to afford 109 (14 mg, 24%) as a brown solid. LCMS: [M+H]+ 668. 1H NMR (500 MHz, DMSO) δ 8.60 (d, J=2.5, 1H), 8.48 (d, J=5.0, 1H), 8.42 (s, 1H), 7.85(d, J=3.0, 1H), 7.44 (d, J=2.0, 1H), 7.34-7.38 (m, 2H), 7.23 (d, J=9.0, 1H), 4.94 (t, J=5.0, 1H), 4.55 (t, J=7.0, 2H), 4.39-4.46 (m, 4H), 4.14-4.19 (m, 1H), 3.79-3.83 (m, 1H), 25 3.59(s, 3H), 3.42-3.44 (m, 1H), 3.00-3.07 (m, 5H), 2.85-2.90 (m, 1H), 2.76 (s, 2H), 2.52-2.59 (m, 2H), 2.36-2.39 (m, 4H), 1.21(d, J=6.5, 6H) Example 110a 1-Methyl(6-(4-methylpiperazinyl)pyridinylamino) (4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 110a A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 30 condenser was charged with 5-bromomethyl(6-(4-methylpiperazinyl)pyridin ylamino)pyridin-2(1H)-one (0.45 g, 1.08 mmol), (PinB)2 (1.37 g, 5.4 mmol), Pd2(dba)3 (49 mg, 0.054 mmol), X-Phos (52 mg 0.11 mmol), KOAc(318 mg, 3.24 mmol), 1, ane 20 mL). After three cycles of vacuum/argon flush, the reaction mixture was heated at 60oC for 15 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under 124 reduced pressure to afford crude 110a, which was used directly in the next on. MS: [M+H]+ 426.
Example 110b 4-(1-Methyl(6-(4-methylpiperazinyl)pyridinylamino)- 6-oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- 5 yl)nicotinaldehyde 110b A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was d with 4-chloro(1-oxo-3,4,6,7,8,9-hexahydropyrazino-[1,2- a]indol-2(1H)-yl)nicotinaldehyde 103b (377mg, 1.15 mmol), 110a (320 mg, 0.78 mmol), Pd(dppf)Cl2 (130 mg, 0.16 mmol), K3PO4.3H2O (52.9 mg, 0.23 mmol), and tetrahydrofuran 10 (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at reflux overnight, cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography g with dichloromethane/methanol (40:1) to afford 110b as yellow solid (351 mg, 76%).
MS: [M+H]+ 593. 15 Example 110 2-(3-(Hydroxymethyl)(1-methyl(6-(4-methylpiperazin idineylamino)oxo-1,6-dihydropyridinyl)pyridinyl)-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-1(2H)-one 110 To the solution of 110b (60 mg, 0.1 mmol) in methanol (50 mL) was added sodium borohydride (11.5 mg, 0.3 mmol) at 0 oC and the mixture was stirred for another 30 minutes. 20 Then the reaction mixture was quenched with water (3 mL) and concentrated. The e was purified with reverse-phase prep-HPLC to afford 110 (26.2 mg, 49%). LCMS: [M+H]+ 595. 1H NMR (500 MHz, CDCl 3) δ 8.69 (d, J = 1.5, 1H), 8.46 (d, J = 5.0, 1H), 7.97 (s, 1H), 7.80 (s, 1H), 7.41-7.37 (m, 1H), 7.34 (d, J = 1.5, 1H), 6.89 (s, 1H), 6.22 (d, J = 8.0, 1H), 6.15 (d, J = 8.5, 1H), 5.10 (t, J = 6.5, 1H), 4.66-4.64 (m, 1H), 4.51-4.30 (m, 2H), 4.15-4.12 (m, 25 2H), 3.93-3.89(m, 1H), 3.71 (s, 3H), 3.58-3.48 (m, 4H), 2.61-2.56(m, 7H), 2.47-2.39 (m, 3H), 1.91-1.87 (m, 2H), 1.79-1.78 (d, J = 5.0, 3H) Example 111a (6-Aminopyridinyl)(morpholino)methanone 111a O N O N NH2 111a To a solution of morpholine (9.00 g, 103 mmol) in EtOH (400 mL) was added EDCI 30 (10.0 g, 52.2 mmol), HOBt (7.00 g 51.8 mmol), and 6-aminonicotinic acid (6.00 g, 43.4 mmol). After stirring for 18 h, the resulting sion was filtered. The solid was triturated 125 with a mixture of MeOH (100 mL) and methylene chloride (100 mL) to afford 111a as a white solid (2.7 g, 30%). LCMS: (M+H)+ 208 Example 111b 5-Bromomethyl(5-(morpholinecarbonyl)pyridin ylamino)pyridine-2(1H)-one 111b O N O N NH O N Br 5 111b Following the procedure described for synthesis of 101i, intermediate 111a and 3,5- dibromomethylpyridin-2(1H)-one were d to give 111b in 21% yield. LCMS: (M+H)+ 394. 1H NMR (500 MHz, MeOD) δ 8.84 (d, J=2.5, 1H), 8.42 (d, J=2, 1H), 7.72 (m, 1H), 7.42 (d, J=2, 1H), 7.11 (d, J=8.5, 1H), 3.72 (m, 8H), 3.63 (s, 3H). 10 Example 111c 1-Methyl(5-(morpholinecarbonyl)pyridinylamino) (4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 111c A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a condenser was charged with 111b (1.0 g, 0.25 mmol), X-phos (120 mg, 0.025 mmol), Pd2(dba)3 (110 g, 0.0125 mmol), KOAc (750 mg, 0.75 mmol), 4,4,4',4',5,5,5',5'-octamethyl- 15 2,2'-bi(1,3,2-dioxaborolane) (3.2g, 1.25mmol) and 1,4-dioxane (50 mL). After three cycles of vacuum/argon flush, the reaction e was heated at 100 oC for 15 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced re and the resulting residue was purified by flash column chromatography eluting with 5:1 petroleum ether/ethyl acetate to afford 111c as a yellow solid (700 mg, 63%). MS: [M+H]+ 441. 20 Example 111d 4-(1-Methyl(5-(morpholinecarbonyl)pyridinylamino)- 1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- yl)nicotinaldehyde 111d A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a condenser was charged with 111c (450 mg, 1.26 mmol), 4-chloro(1-oxo-3,4,6,7,8,9- 25 hexahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 103b (413 mg, 1.26 mmol), f)Cl2 (102 mg, 0.126 mmol), K3PO4.3H2O (85 mg, 0.352 mmol), and THF (10 mL).
After three cycles of vacuum/argon flush, the reaction mixture was heated at 100oC for 15 h.
It was then cooled to room temperature and filtered. The filtrate was trated under reduced pressure and the resulting residue was purified by flash column chromatography 126 eluting with 7:1 petroleum ether/ethyl acetate to afford 111d as a yellow solid (700 mg, 63%).
MS: [M+H]+ 608.
Example 111 2-(3-(Hydroxymethyl)(1-methyl(5-(morpholine carbonyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)-3,4,6,7,8,9- 5 hexahydropyrazino[1,2-a]indol-1(2H)-one 111 A mixture of 111d (60 mg, 0.05 mmol), NaBH4 (6.4mg, 0.1mmol) and MeOH (5 mL) was stirred at 0 oC for 30mins. The mixture was evaporated in vacuo and the residue was ted with EtOAc (10 mL X 2). The combined EtOAc extract was concentrated under reduced pressure and the e was purified with reverse-phase prep-HPLC to give 111 (27 10 mg, 44%). LCMS: [M+H]+ 610. 1H NMR (500 MHz, DMSO) δ 9.00 (s, 1H), 8.78 (d, J=2.0, 1H), 8.49 (d, J=5, 1H), 8.26 (d, J=2.0, 1H), 7.65-7.67 (m, 2H), 7.60 (d, J=2.5, 1H), 6.58 (s,1H), 4.96 (t, J=5, 1H), 4.40-4.46 (m, 2H), .24 (m, 3H), 3.86-3.88 (m, 1H), 3.56-3.62 (m, 8H), 3.50 (s, 4H), 2.62-2.63 (m, 2H), 2.46-2.47 (m, 2H), 1.67-1.80 (m, 4H) Example 112a Methyl 8-Tetrahydroindolizinecarboxylate 112a CO2Me N 15 112a A 500-mL round-bottomed flask equipped with a magnetic stirrer and nitrogen inlet was purged with nitrogen and charged with 5,6,7,8-tetrahydroindolizinecarboxylic acid (30.4 g, 184 mmol), DMF (1.00 g, 13.6 mmol) and methylene chloride (300 mL). The solution was cooled to 0 °C using an ice bath. Oxalyl chloride (28.0 g, 221 mmol) was added 20 dropwise, and the reaction mixture was warmed to room temperature over 30 min and stirred for 5 h. After this time, the resulting on was concentrated to afford a brown solid. This solid was dissolved in anhydrous methanol (400 mL), and the solution was cooled to 0 °C.
Triethylamine (57 g, 552 mmol) was added to the reaction mixture, and it was stirred for a further 2 h at room temperature. After this time, the reaction mixture was concentrated to 25 dryness under reduced pressure. The e was d with methylene chloride (300 mL) and washed with water (200 mL) and saturated aqueous sodium bicarbonate (200 mL). The organic layer was dried over sodium sulfate, ed and concentrated under reduced pressure.
The resulting residue was titrated with hexane (200 mL) to afford 112a in 58% yield (19.1 g) as a white solid: mp 72–74 °C; 1H NMR (300 MHz, DMSO-d6) δ 7.13 (s, 1H), 6.23 (s, 1H), 127 3.93 (t, 2H, J = 6.0 Hz), 3.77 (s, 3H), 2.75 (t, 2H, J = 6.0 Hz), 1.93 (m, 2H), 1.80 (m, 2H); (APCI+) m/z 180.1 (M+H) Example 112b Methyl 3-(Cyanomethyl)-5,6,7,8-tetrahydroindolizine carboxylate 112b CO2Me CN N 5 112b A 500-mL three-neck round-bottomed flask equipped with an addition , thermometer and charged with 112a (6.70 g, 37.4 mmol), Iodoacetonitrile (12.5 g, 74.9 mmol), iron (II) sulfate heptahydrate (5.20 g, 18.7 mmol) and dimethyl sulfoxide (250 mL).
Hydrogen peroxide (35%, 18.2 g, 187 mmol) was added dropwise to the mixture in 1 h 10 through a syringe pump at room ature using a water bath. Iron (II) sulfate heptahydrate (2 to 3 equivalent) was added to the reaction mixture in portions to keep the temperature between 25 oC to 35 oC, until the color of the reaction mixture is deep red. If TLC shows the reaction not completed, then more hydrogen de (2-3 equivalent) and more iron (II) e heptahydrate (1-2 equivalent) are added in the same manner until the 15 reaction is completed. After that time, the reaction mixture was partitioned between saturated sodium onate solution (200 mL) and ethyl acetate (400 mL). The organic layer was separated, and the aqueous layer was ted with ethyl acetate (2 × 100 mL). The combined organic layers were washed with saturated Sodium thiosulfate solution (50 mL), dried over sodium sulfate and concentrated under reduced pressure. The residue was purified 20 by column chromatography to afford a 78% yield (6.40 g) of 112b as a yellow oil: 1H NMR (500 MHz, CDCl3) δ 6.23 (s, 1H), 4.23 (s, 2H), 3.94 (t, 2H, J = 6.5 Hz), 3.81 (s, 3H), 2.74 (t, 2H, J = 6.5 Hz), 2.00 (m, 2H), 1.83 (m, 2H); ) m/z 219.3 (M+H) Example 112c Methyl 3-(2-Aminoethyl)-5,6,7,8-tetrahydroindolizine carboxylate Hydrogen Chloride Salt 112c 25 Methyl 3-(Cyanomethyl)-5,6,7,8-tetrahydroindolizinecarboxylate 112b was hydrogenated with platinum oxide catalyst under 50 psi of hydrogen in ethanol and ethyl 128 acetate in the presence of hydrogen chloride overnight at room temperature to give 112c (380 mg, 1.74 mmol) which was used directly in the next step.
Example 112d 3,4,6,7,8,9-Hexahydropyrido[3,4-b]indolizin-1(2H)-one 112d 5 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and nitrogen inlet was purged with nitrogen and charged with methyl 3-(2-aminoethyl)-5,6,7,8- tetrahydroindolizinecarboxylate hydrogen chloride salt 112c (prepared above, estimated 1.74 mmol, presuming quantitative yield), sodium ethoxide (354 mg, 5.22 mmol) and l (20 mL). The mixture was stirred at 55 oC for 5 h. After that time, the reaction mixture was 10 concentrated under reduced pressure and the residue was partitioned n ethyl acetate (200 mL) and water (100 mL). The organic layer was separated, and the aqueous layer was ted with ethyl e (2 × 100 mL). The combined organic layers were washed with brine, dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography to afford a 67% yield (220 mg) of 112d as a white solid: 15 mp 195–197 °C; 1H NMR (500 MHz, DMSO-d6) δ 6.76 (s, 1H), 5.89 (s, 1H), 3.78 (t, 2H, J = 6.5 Hz), 3.35 (m, 2H), 2.66 (m, 4H),1.87 (m, 2H), 1.72 (m, 2H); ) m/z 191.3 (M+H) Example 112e 3-Bromo(1-oxo-3,4,6,7,8,9-hexahydropyrido[3,4- b]indolizin-2(1H)-yl)isonicotinaldehyde 112e A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and 20 reflux condenser was charged with 1,4-dioxane (15 mL), 3,5-dibromoisonicotinaldehyde (400mg, 1.5 mmol), 112d (142 mg, 0.76 mmol) and cesium carbonate (176 mg, 1.5 mmol). os (40 mg, 0.08 mmol) and Pd2(dba)3 (70 mg, 0.08 mmol) were added, and the reaction mixture was heated at 100 ºC for 5 h. After this time the reaction was cooled to room ature and filtered. The filtrate was concentrated under reduced pressure and the 25 residue was ed on flash column eluting with DCM:MeOH (20:1) to afford 112e (200 mg, 70%). MS: [M+H] Example 112f ethyl(5-(4-(oxetanyl)piperazinyl)pyridin ylamino)oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrido[3,4- b]indolizin-2(1H)-yl)isonicotinaldehyde 112f 30 A sealed tube was charged with 112e (200 mg, 0.53 mmol), 1-methyl(5-(4-(oxetan- 3-yl) piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolan 129 yl)pyridine-2(1H)-one 101l (240 mg, 0.51 mmol), PdCl2(dppf) (42 mg, 0.05 mmol), K3PO4 (230 mg, 1 mmol), and NaOAc (80 mg, 1 mmol) in CH3CN (5 mL) and H2O (1.5 mL). The system was evacuated and refilled with N2.The reaction mixture was heated at 100 ºC for 2 h.
It was then cooled to room temperature and filtered. The filtrate was concentrated under 5 reduced pressure and the resulting e was purified by flash column chromatography eluting with 10:1 of DCM/MeOH to afford 112f (138 mg, 40%) as a pale yellow solid. MS: [M+H]+ 635.
Example 112 2-(4-(Hydroxymethyl)(1-methyl(5-(4-(oxetan yl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 10 3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin-1(2H)-one 112 A mixture of 112f (130 mg, 0.20 mmol) and NaBH4 (20 mg, 0.5 mmol) in MeOH (5 mL) was stirred at 0 oC for 0.5 h. The mixture was quenched with water and extracted with EtOAc (5 mL X 2). The combined EtOAc extract was concentrated under reduced pressure and the residue was purified with reverse-phase prep-HPLC to afford 112 (48 mg, 34%). 15 LCMS: [M+H]+ :637. 1H NMR (500 MHz, DMSO) δ 8.58 , 8.48 (s, 1H), 8.45 (s, 1H), 8.39 (s, 1H), 7.86 (d, J=2.4, 1H), .36 (m, 2H), 7.23-7.21 (m, 1H), 6.02 (s, 1H), 5.02 (s, 1H), 4.54 (t, J=6.5, 2H), 4.45 (t, J=5.5, 2H), 4.36-4.35 (m, 2H), 4.00-3.79 (m, 4H), 3.59 (s, 3H) , 3.43-3.41 (m, 1H), 3.07-2.96 (m, 6H), 2.70 (t, J=6.0, 2H) , 2.39-2.38 (m, 4H), 1.92-1.90 (m, 2H), 1.75-1.73 (m, 2H). 20 Example 113a (3-Nitro-1H-pyrazolyl)methanol 113a O2N OH N N H 113a A 3-L three-neck round-bottomed flask ed with a mechanical stirrer, addition funnel and nitrogen inlet was purged with nitrogen and charged with 3-nitropyrazole carboxylic acid (28.0 g, 178 mmol) and THF (420 mL) and cooled to –5 °C using an 25 ice/acetone bath. Borane-THF complex solution (1.0 M, 535 mL, 535 mmol) was added at a rate that maintained the internal reaction temperature below 5 °C. After the addition was complete the cooling bath was removed and the reaction was stirred at room temperature for 18 h. After this time the reaction was cooled to –5 °C using an ice/acetone bath, water (70 mL) and 4N hloric acid (70 mL) was added and the on was stirred at reflux for 1 30 h in order to destroy the borane x with pyrazole. The reaction was cooled to room temperature and concentrated under reduced pressure to a volume of approximately 30 mL. 130 Ethyl acetate (175 mL) was added and the mixture stirred for 15 min. The aqueous layer was separated and extracted with ethyl acetate (4 × 200 mL). The combined organic layers were washed with saturated aqueous sodium bicarbonate (2 × 50 mL), brine (50 mL) and dried over sodium sulfate, the drying agent was removed by filtration, and the filtrate concentrated 5 under reduced pressure to afford (3-nitro-1H-pyrazolyl)methanol 113a in a 94% yield (24.0 g) as a light yellow solid: 1H NMR (300 MHz, DMSO-d6) δ 13.90 (br s, 1H), 6.87 (s, 1H), 5.58 (t, 1H, J = 5.4 Hz), 4.53(d, 2H, J = 5.1 Hz); MS (ESI+) m/z 144.0 (M+H) Example 113b (1-(2-Bromoethyl)nitro-1H-pyrazolyl)methanol 113b O2N OH N N Br 113b 10 A 1-L three-necked round-bottomed flask equipped with a mechanical stirrer and thermoregulator was purged with en and charged with 113a (25.0 g, 175 mmol), DMF (250 mL), and cesium carbonate (70.0 g, 215 mmol) was heated at 104 °C for 5 min. The reaction e was then cooled to 0 °C using an ice/acetone bath and dibromoethane (329 g, 1.75 mol) was added portionwise (no rm). The reaction was stirred at 0 °C for 1 then 15 at room ature for 4 h. After this time a on of KH2PO4 (40 g) in water (400 mL) was added slowly. The reaction mixture stirred at room temperature for 30 min. Ethyl acetate (450 mL) was added and the aqueous layer was separated and extracted with ethyl acetate (2 × 100 mL). The combined c layers were washed with water (200 mL), brine (200 mL), dried over sodium sulfate, and the drying agent was removed by filtration. The 20 filtrate was concentrated under reduced pressure to afford an 86% yield (37.5 g) of crude 113b as an orange oil: 1H NMR (300 MHz, CDCl3) δ 6.85 (s, 1H), 4.82 (d, 2H, J = 5.4 Hz), 4.66 (t, 2H, J = 6.3 Hz), 3.83 (t, 2H, J = 6.3 Hz); MS (ESI+) m/z 249.9 (M+H).
Example 113c 1-(2-Bromoethyl)(bromomethyl)nitro-1H-pyrazole 113c O2N Br N N Br 113c 25 A 500-mL three-necked round-bottomed flask equipped with a ic stirrer, nitrogen inlet and reflux condenser was purged with en and charged with 113b (37.0 g, 148 mmol) and chloroform (160 mL). The reaction was cooled to –5 °C using an ice/acetone bath and phosphorous tribromide (40.0 g, 148 mmol) was added portionwise. The cooling 131 bath was removed and the reaction stirred at reflux for 2 h. After this time, the reaction was cooled to –5 °C and saturated aqueous sodium bicarbonate (250 mL) was added until a pH of 8.5 was reached. The e was extracted with ethyl acetate (3 × 150 mL) and the combined organic layers were washed with saturated aqueous sodium carbonate (2 × 50 mL), 5 brine (75 mL), dried over sodium sulfate and the drying agent was removed by filtration. The filtrate was concentrated under reduced pressure to afford a yellow residue that was dissolved with gentle heating in methylene chloride (60 mL). Hexanes (approximately 20 mL) was added and the solution became cloudy. The mixture was heated until a solid precipitate formed, ene de (9 mL) was added and the solution became clear. The solution 10 was left to cool to room temperature and after 4 h the resulting crystals were collected by vacuum filtration. The filter cake was washed with a ice cold 1:2 mixture of ene chloride:hexanes (2 × 20 mL) to afford 1-(2-bromoethyl)(bromomethyl)nitro-1H- pyrazole (19.7 g). The combined filtrates were ated and the procedure was performed again to afford an additional 9.70 g of 1-(2-bromoethyl)(bromo-methyl)nitro-1H- 15 pyrazole. The solids were combined and dried under high vacuum for 18 h to afford a 57% yield (26.0 g) of 1-(2-bromoethyl)(bromomethyl)nitro-1H-pyrazole 113c as white crystals: mp 95–97 °C; 1H NMR (300 MHz, CDCl3) δ 6.93 (s, 1H), 4.63 (t, 2H, J = 6.0 Hz), 4.54 (s, 2H), 3.86 (t, 2H, J = 6.0 Hz).
Example 113d 5-Methylnitro-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine 20 113d O2N N N N CH3 113d A 1-L single-neck round-bottomed flask equipped with a magnetic stirrer and nitrogen inlet was charged with THF (350 mL), 113c (10.0 g, 32.2 mmol), 2M methylamine solution in THF (113 mL, 225 mmol) and stirred at room temperature for 72 h. After this 25 time the reaction was concentrated to dryness under reduced pressure, and the ing solid was stirred with a mixture of ethyl e (75 mL) and 10% aqueous potassium ate (75 mL). The aqueous layer was separated and extracted with ethyl acetate (2 × 75 mL). The ed organic ts were washed with 10% aqueous potassium carbonate (75 mL), followed by brine (50 mL) and dried over sodium sulfate. The drying agent was removed by 30 filtration, and the filtrate concentrated under reduced pressure to afford 113d in 97% yield 132 (5.70 g) as a yellow solid: 1H NMR (300 MHz, CDCl3) δ 6.62 (s, 1H), 4.28 (t, 2H, J = 5.4 Hz), 3.67 (s, 2H), 2.95 (t, 2H, J = 5.4 Hz), 2.52 (s, 3H); MS (ESI+) m/z 183.0 (M+H) Example 113e 5-Methyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinamine 113e H2N N N N CH3 5 113e A 500-mL Parr reactor bottle was purged with nitrogen and d with 10% ium on carbon (50% wet, 800 mg dry weight) and a solution of 113d (4.00 g, 2.20 mmol) in ethanol (160 mL). The bottle was attached to Parr hydrogenator, evacuated, charged with hydrogen gas to a pressure of 45 psi and shaken for 2 h. After this time, the 10 en was evacuated, and nitrogen was charged into the bottle. CELITE® 521 (1.0 g) was added, and the mixture was filtered through a pad of CELITE® 521. The filter cake was washed with ethanol (2 × 75 mL), and the combined filtrates were concentrated to dryness under reduced pressure to afford a 99% yield of 113e (3.31 g) as an orange solid: 1H NMR (300 MHz, CDCl3) δ 5.34 (s, 1H), 3.98 (t, 2H, J = 5.4 Hz), 3.52 (s, 3H), 2.84 (t, 2H, J = 5.7 15 Hz), 2.45 (s, 3H); MS (ESI+) m/z 153.1 (M+H) Example 113f 5-Bromomethyl(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazin ylamino) pyridin-2(1H)-one 113f N N N NH O N Br 113f A sealed tube equipped with a magnetic stirrer was charged with 113e (1.02 g, 6.7 20 mmol), 3,5-dibromomethylpyridin-2(1H)-one (2.15 g, 8.1 mmol), Pd2(dba)3 (610 mg, 0.67mmol), 2,2-bis(diphenylphosphino)-1,1-binaphthyl (775 mg, 1.34 mmol), cesium carbonate (4.37 g, 13.6 mmol), and 1,4-dioxane (30 mL). After three cycles of /argon flush, the mixture was heated at 110oC for 2 h. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified by silica gel column chromatography g 25 with dichloromethane/methanol (15:1, V/V) to afford 113f (380 mg, 14%) as a white solid.
LCMS: [M+H]+ 338 133 Example 113g 2-(4-chloro(hydroxymethyl)pyridinyl)-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-1(2H)-one 113g To a solution of 4-chloro(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- 5 yl)nicotinaldehyde 103b (1.0 g, 3.0 mmol) in methanol (50 mL) was added sodium borohydride (380 mg, 9.0 mmol) at 10 oC and the mixture was stirred for another 30 minutes.
Then the reaction mixture was quenched with water (1 mL) and concentrated. The residue was dissolved in dichloromethane (50 mL) and washed with water (10 mL). The organic phase was dried over anhydrous Na2SO4, filtered, and evaporated under d pressure to 10 afford 113g as a yellow solid (900 mg, 90%). MS: [M+H]+ 332.
Example 113h (4-Chloro(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- idineyl)methyl acetate 113h To a mixture of 113g (900 mg, 2.7 mol) and triethylamine (900 mg, 9.0 mol) in 15 dichloromethane (5 mL) was added se acetyl chloride (600 mg, 6.0 mol) while stirring at room temperature and stirred for another 1 h. The reaction mixture was concentrated and purified by silica-gel column chromatography eluting with dichloromethane to afford 113h as white solid (950 mg, 94%). MS: [M+H]+ 374.
Example 113i (2-(1-Oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- 20 (4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridinyl)methyl acetate 113i A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with 113h (950 mg, 2.5 mmol), Pin2B2 (1.6 g, 2.0 eq., 5 mmol), 134 Pd2(dba)3 (230 mg, 0.1 eq., 0.25 mmol), X-phos (232 mg, 0.2 eq., 0.5 mmol), AcOK (735 mg, 3 eq., 7.5 mmol) and dioxane (20 mL). After three cycles of /argon flush, the mixture was heated to 65 ºC for 14 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was washed by 5 PE/EA=3/1 (10 mL) to afford 113i as yellow solid (950 mg, 87%). MS: [M+H]+ 383. e 113j (4-(1-Methyl(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinylamino)oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl acetate 113j N N N NH AcO O N N N O N 113j 10 A sealed tube equipped with a magnetic stirrer was charged with 113f (190 mg, 0.56 mmol), 113i (215 mg, 0.56 mmol), Pd Cl2 (47 mg, 0.056 mmol), 1.0 M NaOAc (93 mg, 1.12 mmol, 2.0 equiv), 1.0 M K3PO4 (240 mg, 1.12 mmol, 2.0 equiv), and acetonitrile (3 mL).
After three cycles of vacuum/argon flush, the mixture was heated at 110oC for 2 h. It was then ed and the filtrate was evaporated in vacuo. The residue was purified by silica gel 15 column chromatography eluting with dichloromethane/methanol (10:1, V/V) to afford 113j (300 mg, 94%) as a brown solid. LCMS: [M+H]+ 597 Example 113 2-(3-(Hydroxymethyl)(1-methyl(5-methyl-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 113 20 A mixture of 113j (300 mg, 0.50 mmol) and LiOH.H2O , 2.50 mmol) in iPrOH/THF (1:1, 3 mL) and H oC for 2 h. The mixture was 2O (1 mL) was stirred at 30 evaporated in vacuo and the residue was extracted with EtOAc (10 mL X 2). The combined EtOAc extract was concentrated under reduced pressure and the residue was purified with reverse-phase prep-HPLC to afford 113 (91 mg, 32%) as a white solid. LCMS: [M+H]+ 555. 25 1H NMR (500 MHz, CDCl 3) δ 8.47 (d, J=5.0, 1H), 7.95 (d, J=5.0, 1H), 7.72 (d, J=2.0, 1H), 7.42 (s, 1H), 7.35 (d, J=5.0, 1H), 6.89 (s, 1H), 5.69 (s,1H), 5.01-5.02 (m, 1H), .62 (m, 1H), 4.48-4.49 (m, 1H), 4.32-4.33 (m, 1H), 4.15-4.07 (m, 4H), 3.86-3.87 (m, 1H), 3.69 (s, 135 3H), 3.60-3.59 (m, 2H), 2.88 (t, J=6.0, 2H), 2.61-2.56 (m, 4H), 2.47 (s, 3H), 1.89-1.90 (m, 2H), 1.78-1.79 (m, 2H) Example 114a (R)bromo(4-(1,4-dimethyloxopiperazin yl)phenylamino)methylpyrazin -2(1H)-one 114a O N N HN O N N Br 5 114a A sealed tube equipped with a magnetic stirrer was charged with (R)(4- henyl)-1,4-dimethylpiperazinone (1.08 g, 5 mmol), 3,5-dibromomethylpyridin- 2(1H)-one (1.47 g, 5.5 mmol), diisopropylethylamine (1.94 g, 15 mmol), and iPrOH (20 mL).
After three cycles of vacuum/argon flush, the mixture was heated at 110 oC ght. After 10 cooling down to room ature, water (20 mL) was added to, and the mixture was extracted with ethyl acetate (50 mL X 2). The organic layer was separated, combined, dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by silica gel column chromatography eluting with dichloromethane/methanol (10:1, V/V) to afford 114a (1.8 g, 90%) as a red solid. LCMS: [M+H]+ 406 15 Example 114b (R)-(4-(6-(4-(1,4-dimethyloxopiperazinyl)phenylamino)- 4-methyloxo-4,5-dihydropyrazinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)pyridinyl)methyl acetate 114b A sealed tube equipped with a magnetic stirrer was charged with 114a (228 mg, 0.56 20 mmol), 3-(acetoxymethyl) (1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- idinylboronic acid 113i (215 mg, 0.56 mmol), Pd (dppf)Cl2 (47 mg, 0.056 mmol), 1.0 M NaOAc (93 mg, 1.12 mmol, 2.0 equiv), 1.0 M K3PO4 (240 mg, 1.12 mmol, 2.0 equiv), and acetonitrile (3 mL). After three cycles of vacuum/argon flush, the mixture was heated at 136 110oC for 2 h. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified by silica gel column chromatography eluting with dichloromethane/methanol (10:1, V/V) to 114b (360 mg, 96%) as a brown solid. LCMS: [M+H]+ 665.
Example 114 (R)(4-(6-(4-(1,4-dimethyloxopiperazin 5 yl)phenylamino)methyloxo-4,5-dihydropyrazinyl)(hydroxymethyl)pyridinyl)- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 114 A mixture of 114b (360 mg, 0.54 mmol) and LiOH.H2O (138mg, 2.76 mmol) in iPrOH/THF (1:1, 3 mL) and H oC for 2 h. The mixture was 2O (1 mL) was stirred at 30 evaporated in vacuo and the residue was extracted with EtOAc (10 mL x 2). The combined 10 EtOAc extract was concentrated under reduced re and the e was purified with reverse-phase prep-HPLC to afford 114 (72 mg, 21%) as a white solid. LCMS: [M+H]+ 623. 1H NMR (500 MHz, CDCl 3) δ 8.53 (d, J=4.5, 1H), 8.31 (s, 1H), 8.10 (s, 1H), 7.83-7.78 (m, 3H), 7.36 (d, J=8.0, 2H), 6.89 (s, 1H), .14 (m, 1H), 4.68-4.70 (m, 1H), 4.49-4.53 (m, 1H), 4.38-4.43 (m, 1H), 4.15-4.06 (m, 2H), 3.89-3.90 (m, 1H), 3.72-3.73 (m, 2H), 3.65 (s, 15 3H), 3.21-3.22 (m, 1H), 3.01-3.03 (m, 4H), 2.71-2.56 (m, 5H), 2.20 (s, 3H), 1.90-1.92 (m, 2H), 1.79-1.80 (m, 2H) Example 115a 5-Bromomethyl(5-methyl-1H-pyrazolylamino)pyridin- 2(1H)-one 115a HN N NH O N Br 115a 20 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and reflux condenser was charged with 1,4-dioxane (15 mL), yl-1H-pyrazolamine (1 g, 10 mmol) (1), 3,5-dibromomethylpyridin-2(1H)-one (4 g, 15 mmol) (2), and cesium carbonate (6.4 g, 20 mmol). Xantphos (400 mg, 0.8 mmol) and a)3 (700 mg, 0.8 mmol) were added, and the reaction e was heated at 100 ºC for 5 h. After this time the 25 on was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the residue was purified on flash column eluting with DCM:MeOH (20:1) to afford 115a (1.0 g, 35%). MS: [M+H]+ 283. 137 Example 115b 4-(1-Methyl(5-methyl-1H-pyrazolylamino)oxo-1,6- dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- yl)nicotinaldehyde 115b 5 A sealed tube was d with 115a (280 mg, 1 mmol), 3-(acetoxymethyl)(1-oxo- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinylboronic acid 113i (420 mg, 1.1 mmol), PdCl2(dppf) (41 mg, 0.056 mmol), K3PO4 (100 mg), and NaOAc (50 mg) in CH3CN (10 mL) and H2O (3 mL). The system was evacuated and refilled with N2. The reaction mixture was heated at 100 ºC for 2 h. It was then cooled to room temperature and 10 filtered. The filtrate was concentrated under reduced re and the ing residue was purified by flash column chromatography eluting with 10:1 of DCM/MeOH to afford 115b in 35% yield (190 mg) as a pale yellow solid. MS: [M+H]+ 542.
Example 115 2-(3-(Hydroxymethyl)(1-methyl(5-methyl-1H-pyrazol ylamino)oxo-1,6-dihydropyridinyl)pyridinyl)-3,4,6,7,8,9-hexahydropyrazino[1,2- 15 a]indol-1(2H)-one 115 A 100-mL single-neck round-bottomed flask was charged with 115b (190 mg, 0.35mol) in THF/iPA/H2O (5 mL/5 mL/2 mL) and LiOH (85 mg, 3.5 mmol) while stirring.
This mixture was stirred at 50 ºC for 0.5 h. Then 20 mL H2O was added and the mixture was ted with EA (30 mL X 3). The combined c layer was dried over Na2SO4 and 20 concentrated to give a yellow solid, which was further purified by reverse-phase prep-HPLC to afford 115 as a white solid (48 mg, 30% yield). LCMS: [M+H]+ 500. 1H NMR (500 MHz, CDCl3) δ 8.44 (d, J=6.0, 1H), 7.95 (s, 1H), 7.69 (s, 1H), 7.44 (s, 1H), 7.30 (d, J=6.0, 2H), 6.87 (s, 1H), 5.74 (s, 1H), 4.59-3.86 (m, 7H), 3.69 (s, 3H), 2.57-2.56 (m, 4H) , 2.25 (s, 3H) 1.88-1.77 (m, 4H) 25 Example 116a 3-Bromo{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca- 1(9),2(7),3-trienyl}pyridinecarbaldehyde 116a 138 To a 100-mL single-neck round-bottomed flask equipped with a ic stirrer and reflux condenser was charged with 1,4-dioxane (15 mL), 3,5-dibromoisonicotinaldehyde (200 mg, 0.76 mmol), 8-thia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienone 191d (160 5 mg, 0.76 mmol), and cesium carbonate (176 mg, 1.5 mmol). Cuprous iodide CuI (100 mg, 0.76 mmol) and 4,7-dimethoxy-1,10-phenanthroline (127 mg, 0.52 mmol) were added, and the reaction mixture was heated at 100 ºC for 5 h. After this time the reaction was cooled to room temperature and filtered. The filtrate was trated under d pressure and the residue was purified on flash column eluting with EtOAC/PE (1:2) to afford 116a (80 mg, 10 30%). MS: [M+H]+ 390.
Example 116b 3-[1-Methyl({5-[4-(oxetanyl)piperazinyl]pyridine yl}amino)oxo-1,6-dihydropyridinyl]{6-oxothia-4,5- diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridinecarbaldehyde 116b 15 A sealed tube was charged with 116a (80 mg, 0.20 mmol), 1-methyl(5-(4-(oxetan- 3-yl)piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolan yl)pyridin-2(1H)-one 101l (96 mg, 0.20 mmol), PdCl2(dppf) (18 mg, 0.02 mmol), K3PO4 (30 mg), and NaOAc (20 mg) in CH3CN (5 mL) and H2O (1 mL). The system was evacuated and refilled with N2. The reaction mixture was heated at 100 ºC for 2 h. It was then cooled to 20 room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by flash column tography eluting with 10:1 of DCM/MeOH to afford 116b in 35% yield (46 mg). MS: [M+H]+ 651.
Example 116 4-Hydroxymethyl[1-methyl({5-[4-(oxetanyl)piperazin- yridineyl}amino)oxo-1,6-dihydropyridinyl]{6-oxothia-4,5- 25 diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridine 116 139 To a on of 116b (46 mg, 0.07 mmol) at 0oC in methanol (4 mL) was added sodium borohydride (20 mg, 0.7 mmol) and stirred for 30 minutes. Then the reaction e was quenched with water (1.0 mL) and concentrated. The residue was purified by reversephase prep-HPLC to afford 116 (12 mg, 28 %) as a yellow solid. LCMS: [M+H]+ 653. 1H 5 NMR (500 MHz, DMSO) δ 8.60 (s,1H), 8.59 (s, 1H), 8.56 (d, J=2.0 ,1H), 8.50 (s, 1H), 8.44 (s, 1H), 7.87 (d, J=3.0, 1H), 7.38-7.36 (m, 2H), 7.24-7.22 (m, 1H), 4.90 (m, 1H), 4.56-4.53 (m, 2H), 4.46-4.44 (m, 4H), 3.59 (s, 3H), 3.44-3.42 (m, 1H), 3.06 (t, J=4.5, 4H), 2.94-2.93(m, 2H), 2.85-2.84 (m, 2H), 2.38(t, J=4.0, 4H), 1.89-1.84 (m, 4H) Example 117a 5-(Methylthio)nitropyridine 117a 10 To a mixture of 5-chloronitropyridine (3 g, 18 mmol) in MeOH (20 mL), sodium methanethiolate (1.4 g, 20 mmol) was added at 0 oC and the mixture stirred at 20 ºC for 2 hours. The resulting suspension was filtered and washed with water, and dried in vacuum to afford crude 117a as a yellow solid (2 g, 66%) without purification for next step. MS: 15 [M+H]+ 171.
Example 117b 5-(Methylsulfonyl)nitropyridine 117b O O S N NO2 117b To a mixture of 117a (260 mg, 0.5 mmol) in acetic acid (15 mL) was added H2O2 (aq. 30%) (7.5 mL) and the reaction mixture was stirred overnight at 25 oC. The reaction solution 20 was poured into water and extracted with EtOAC and trated to a pale yellow , purified by silica gel with (EtOAC/PE:1:3) to give 117b (2 g, 86%). MS: [M+H]+ 203.
Example 117c 5-(Methylsulfonyl)pyridinamine 117c O O S N NH2 117c A mixture of 117b (2 g, 10 mmol), MeOH (10 mL), Pd/C (120 mg) in methanol (8 25 mL) was stirred f at 25 ºC under H2 (50 Psi) ght. The Pd/C was removed by filtration 140 and the filtrate was concentrated under reduced pressure to give 117c (1.7 g, 98%). MS: [M+H]+ 173. e 117d 5-Bromomethyl(5-(methylsulfonyl)pyridin ylamino)pyridin-2(1H)-one 117d O O S N NH O N Br 5 117d A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and reflux condenser was charged with 1,4-dioxane (15 mL), 117c (1.7 g, 10 mmol), 3,5- dibromomethylpyridin-2(1H)-one (5.2 g, 20 mmol) and cesium carbonate (6.4 g, 20 mmol). Xantphos (300 mg, 0.8 mmol) and Pd2(dba)3 (500 mg, 0.8 mmol) were added, and the 10 reaction e was heated at 100 ºC for 5 h ). After this time the reaction was cooled to room temperature. The mixture was removed by filtration and the filtrate was concentrated under reduced pressure. The residue was purified on flash column eluting with DCM:MeOH (20:1) to afford 117d(1 g, 30%). MS: [M+H]+ 358.
Example 117e (4-(1-Methyl(5-(methylsulfonyl)pyridinylamino)oxo- 15 1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- yl)pyridineyl)methyl acetate 117e A sealed tube was charged with 117d (100 mg, 0.28 mmol), 3-(acetoxymethyl)(1- oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinylboronic acid 113i (115 20 mg, 0.3 mmol), PdCl2(dppf) (25 mg, 0.03 mmol), K3PO4 (126 mg, 0.6 mmol), and NaOAc (60 mg, 0.6 mmol) in MeCN (8 mL) and H2O (1 mL). The system was ted and refilled with N2. The reaction mixture was heated at 100 ºC for 2 h. It was then cooled to room temperature and ed. The filtrate was concentrated under reduced pressure and the 141 resulting residue was purified by silica gel flash column eluting with DCM:MeOH (20:1) to afford 117e (100 mg, 40%). MS: [M+H]+ 617.
Example 117 2-(3-(hydroxymethyl)(1-methyl(5- (methylsulfonyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)-3,4,6,7,8,9- 5 hexahydro-pyrazino[1,2-a]indol-1(2H)-one 117 A 100-mL single-neck round-bottomed flask compound was charged with 117e (100 mg, 0.2 mol) in THF/iPA/H2O (5 mL/5 mL/2 mL) and LiOH (50 mg, 2 mmol) while stirring.
This mixture was d at 50 ºC for 0.5 h. Then 20 mL H2O was added and the mixture was extracted with EA (30 mL X 3). The combined organic layer was dried over Na2SO4 and 10 concentrated to give a yellow solid, which was further ed by reverse-phase prep-HPLC to afford 117 as a white solid (72 mg, 90% yield). MS: [M+H]+ 575. 1H NMR (500 MHz, CDCl3) δ 9.39 (s, 1H), 8.84 (d, J=2.0,1H), 8.60 (d, J=2.5, 1H), 8.50 (d, J=2.5, 1H), 7.98 (dd, J=2.5, 4.0, 1H), 7.69 (d, J=2.4, 1H), .47 (d, J=9.0, 1H), 7.38-7.37 (m, 1H), 6.58 (s, 1H), 4.99 (t, J=4.5, 1H), 4.47-4.39 (m, 2H), 4.26-4.11 (m, 3H), .86 (m, 1H), 3.62 (s, 3H), 15 3.19 (s, 3H), 2.66-2.54 (m, 2H), 2.48-2.46 (m, 2H) , 1.79-1.66 (m, 4H) Example 118a tert-Butyl 5-Aminocyclopropyl-1H-pyrazolecarboxylate 118a To a mixture of 3-cyclopropyl-1H-pyrazolamine (0.25 g, 2 mmol) and K2CO3 20 (0.828 g, 6 mmol) in THF (5 mL) was added (Boc)2O (0.436g, 2 mmol) in THF (5 mL). The reaction mixture was stirred at room temperature for 15 h. It was then filtered and concentrated. The residue was purified by flash column eluting with 6:1 petroleum ether/ethyl acetate to afford 118a as a white solid (240 mg, 54%). LCMS: )+ 124.
Example 118b 5-Bromo(3-cyclopropyl-1H-pyrazolylamino) 25 methylpyridin-2(1H)-one 118b N NH HN O N Br 118b 142 A 100-mL single-neck bottomed flask equipped with a magnetic stirrer and reflux condenser was charged with 1,4-dioxane (15 mL), 118a (455 mg, 1.95 mmol), 3,5- dibromomethylpyridin-2(1H)-one (0.40 g, 1.5 mmol), and cesium carbonate (1.22 g, 3.75 mmol). After bubbling nitrogen h the resulting e for 30 minutes, XantPhos (87 5 mg, 0.15 mmol) and tris(dibenzylideneacetone)dipalladium(0) (70 mg, 0.075 mmol) were added, and the reaction e was heated at reflux for 15 h. After this time the reaction was cooled to room temperature, partitioned between ethyl acetate (30 mL) and water (30 mL).
The s layer was separated and extracted with ethyl acetate (50 mL × 2). The organic layers were combined, washed with brine (50 mL) and dried over sodium sulfate. The drying 10 agent was removed by filtration and the filtrate was concentrated under reduced pressure. The residue was purified on flash column eluting with 50:1 DCM/MeOH to afford 118b as a yellow solid (320 mg, 50%). LCMS: (M+H)+ 309. 1H NMR (500 MHz, DMSO) δ 11.85 (s, 1H), 8.23 (s, 1H), 8.02 (d, J = 2.5, 1H), 7.35 (d, J = 2.5, 1H), 5.77 (d, J = 2, 1H), 3.46 (s, 3H), 1.83 (m, 1H), 0.90 (m, 2H), 0.64 (m, 2H) 15 Example 118c (4-(5-(5-Cyclopropyl-1H-pyrazolylamino)methyloxo- 1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- yl)pyridineyl)methyl acetate 118c A sealed tube equipped with a magnetic stirrer was charged with 118b (310 mg, 1 20 mmol), 3-(acetoxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- yl)pyridineylboronic acid 113i (385 mg, 1 mmol), Pd(dppf)Cl2 (80 mg, 0.1 mmol), K3PO4 (424 mg, 2 mmol), NaOAc (165 mg, 2 mmol), CH3CN (15 mL), and water (1 mL). After three cycles of vacuum/argon flush, the mixture was heated at 110 oC for 3 h. It was evaporated in vacuo. The residue was purified by silica gel column tography eluting 25 with dichloromethane/methanol (50:1, V/V) to afford 118c (400 mg, 68%) as a yellow solid.
LCMS: [M+H]+ 569 Example 118 2-(4-(5-(5-Cyclopropyl-1H-pyrazolylamino)methyl oxo-1,6-dihydropyridinyl)(hydroxymethyl)pyridinyl)-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-1(2H)-one 118 143 A mixture of 118c (350 mg, 0.62 mmol) and LiOH.H2O (260 mg, 6.2 mmol) in iPrOH/THF (1:1, 3 mL) and H oC for 1 h. The mixture was 2O (1 mL) was stirred at 30 evaporated in vacuo and the residue was extracted with EtOAc (10 mL X 2). The combined EtOAc extract was concentrated under reduced pressure and the residue was purified by 5 reverse-phase prep-HPLC to afford 118 (200 mg, 54%) as a white solid. LCMS: [M+H]+ 526. 1H NMR (500 MHz, DMSO) δ 11.83 (s, 1H), 8.48 (d, J=5, 1H), 8.05 (d, J=2, 1H), 8.03 (s, 1H), 7.38 (d, J=2, 1H), 7.31 (d, J=5, 1H), 6.58 (s, 1H), 5.81 (d, J=2, 1H), 4.95 (t, J=5, 1H), 4.49-4.51 (m, 1H), 4.38-4.40 (m, 1H), 4.19-4.21 (m, 3H), 3.85-3.87 (m, 1H), 3.58 (s, 3H), .62 (m, 1H), 2.56-2.57 (m, 1H), 2.48-2.49 (m, 2H), 1.81-1.82 (m, 3H), 1.70-1.71 (m, 10 2H), 0.88-0.89 (m, 2H), 0.63-0.64 (m, 2H) Example 119a (S)-(4-(1-Methyl(5-(2-methyl(oxetanyl)piperazin yl)pyridinylamino)oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl acetate 119a O N N N NH AcO N O N N O N 119a 15 Following the procedures as described for 118c, (2-(1-oxo-3,4,6,7,8,9- dropyrazino[1,2-a]indol-2(1H)-yl)(4,4,5,5-tetramethyl-1,3,2-dioxaborolan yl)pyridinyl)methyl e 113i (250 mg) and (S)bromomethyl(3-methyl(4- (oxetanyl)piperazinyl)pyridinylamino)pyridin-2(1H)-one 130e (233 mg) were reacted to give 119a as a yellow solid (230 mg, 62%). LCMS: [M+H]+ 693 20 Example 119 (S)(3-(Hydroxymethyl)(1-methyl(5-(2-methyl(oxetan yl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 119 Following the procedures as described for 118, acetate ysis of 119a with LiOH.H2O in iPrOH/THF (1:1) and H2O, gave 119 as a white solid (184 mg, 85%). LCMS: 25 [M+H]+ 651. 1H NMR (500 MHz, CDCl3) δ 8.65 (d, J=2.5, 1H), 8.50 (d, J=5.0, 1H), 7.99 (s, 1H), 7.87 (s, 1H), 7.84 (d, J=2.0, 1H), 7.35 (d, J=5.0, 1H), 7.33 (d, J=7.0, 1H), 6.90 (s, 1H), 6.83 (d, J=9.0, 1H), 5.04-5.06 (m, 1H), 4.62-4.73 (m, 5H), 4.51 (s, 1H), 4.32 (s, 1H), 4.16 (s, 144 1H), 4.11 (s, 1H), 3.89 (s, 1H), 3.72 (s, 3H), 3.57 (t, J=6.0, 1H), 3.48 (s, 1H), 3.07-3.12 (m, 2H), 2.53-2.63 (m, 7H), 2.24 (m,1H), 1.88-1.93 (m, 2H), 1.80 (s, 2H), 0.99 (d, J=6.5, 3H).
Example 120a o(5-(4-(2-hydroxymethylpropyl)piperazin yl)pyridinylamino)methylpyridin-2(1H)-one 120a HO N N N NH O N Br 5 120a A sealed tube equipped with a magnetic stirrer was charged with 5-bromomethyl- 3-(5-(piperazinyl)pyridinylamino)pyridin-2(1H)-one 101j (500 mg, 1.37 mmol), 2,2- dimethyloxirane (990 mg, 13.7 mmol), Cs2CO3 (1.3 g, 4.11 mmol), and CH3CN (15 mL).
After three cycles of vacuum/argon flush, the e was heated at 110oC for 15 h. It was 10 then filtered and the filtrate was evaporated in vacuum. Crude 120a thus obtained was used in the next step without further purification (460 mg, 77%). LCMS: [M+H]+ 437.
Example 120b (4-(5-(5-(4-(2-Hydroxymethylpropyl)piperazinyl)pyridin ylamino)methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl acetate 120b HO N N N NH AcO O N N N O N 15 120b Following the procedures as described for preparation of 118c, on of 120a (435 mg, 1.0 mmol) and 3-(acetoxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl)pyridinylboronic acid 113i (383 mg, 1 mmol) gave 120b (437 mg, 63%). LCMS: [M+H]+ 696. 145 Example 120 2-(4-(5-(5-(4-(2-Hydroxymethylpropyl)piperazin yl)pyridinylamino)methyloxo-1,6-dihydropyridinyl)(hydroxymethyl)pyridin 4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 120 Following the procedures as described for the preparation of 118, acetate hydrolysis 5 of 120b (70 mg, 0.1 mmol) with LiOH.H2O in iPrOH/THF (1:1) and H2O, gave 120 (27 mg, 42%) as a gray solid. LCMS: [M+H]+ 653. 1H NMR (500 MHz, DMSO-d6) δ 8.61 (d, J=3, 1H), 8.50 (d, J=5, 1H), 8.41 (s, 1H), 7.83 (d, J=3, 1H), 7.46 (d, J=2, 1H), 7.36 (m, 2H), 7.24 (d, ), 6.58 (s, 1H), 4.95 (m, 1H), 4.44 (m, 2H), 4.24 (m, 2H), 4.13 (m, 2H), 3.87-3.88 (m, 1H), 3.60 (s, 3H), 3.03-3.05 (m, 4H), 2.64-2.66 (m, 5H), 2.61-2.63 (m, 1H), 2.49-2.51 (m, 10 2H), 2.24 (s, 2H), 1.70-1.71 (m, 4H), 1.10 (s, 6H).
Example 121a 4-(1-Methyl(5-(4-(oxetanyl)piperazinyl)pyridin ylamino)oxo-1,6-dihydropyridinyl)(1-oxo-6,7,8,9-tetrahydropyrazino[1,2-a]indol- 2(1H)-yl)nicotinaldehyde 121a A flask was charged with 4-chloro(1-oxo-6,7,8,9-tetrahydropyrazino[1,2-a]indol- 15 2(1H)-yl)nicotinaldehyde 103b (88 mg, 0.27 mmol), 1-methyl(5-(4-(oxetan yl)piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin- 2(1H)-one 101l (125 mg, 0.27 mmol), PdCl2(dppf) (18 mg, 0.02 mmol), K3PO4 (30 mg), in THF (5 mL) and H2O (1 mL). The system was evacuated and refilled with N2. The reaction mixture was ed for 4 h, and then cooled to room temperature. It was then filtered and 20 the filtrate was concentrated under d pressure. The resulting residue was purified by flash column chromatography eluting with 10:1 of DCM/MeOH to afford 121a (90 mg, 56%) as a yellow solid. MS: [M+H]+ 633.
Example 121 2-(3-(hydroxymethyl)(1-methyl(5-(4-(oxetan yl)piperazinyl)pyridineylamino)oxo-1,6-dihydropyridinyl)pyridinyl)-6,7,8,9- 25 tetrahydropyrazino[1,2-a]indol-1(2H)-one 121 At 0 oC, to a suspension of 121a (76 mg, 0.12 mmol) in methanol (4 mL) was added sodium borohydride (20 mg, 0.7 mmol) and stirred for 30 minutes. Then the reaction mixture was quenched with water (1.0 mL) and concentrated. The residue was purified by ephase prep-HPLC to afford 121 (56 mg, 74 %). LCMS: [M+H]+ 635. 1H NMR (500 MHz, 30 DMSO) δ 8.66 (d, J=2.0, 1H), 8.57 (d, J=5.0, 1H), 7.93 (d, J=3.0, 1H), 7.85 (d, J=2.5, 1H), 7.80 (s, 1H), 7.50 (d, J=5.0, 1H), 7.24-7.27 (m, 1H), 7.06 (s, 1H), 6.97 (d, J=6.0, 1H), 6.81 (d, J=8.0, 1H), 6.67 (d, J=6.0, 1H), 5.08 (d, J=11.5, 1H), 4.67-4.72 (m, 4H), 4.51 (d, J=12.0, 1H), 4.35 (t, , 1H), 3.72 (s, 3H), 3.57-3.59 (m, 1H), 3.16-3.17 (m, 4H), 2.70-2.74 (m, 4H), .53 (m, 4H), 1.94-1.95 (m, 2H), 1.84-1.86(m, 2H). 146 e 122a (2R, 5S)-tert-Butyl 2,5-Dimethyl(6-nitropyridin yl)piperazinecarboxylate 122a Boc N N N NO2 122a Following the procedures as described for compound 101g, (2R, 5S)-tert-butyl-2,5- 5 dimethylpiperazinecarboxylate (1.5 g, 6.0 mmol), and 5-bromonitropyridine (1212 mg, 6.0 mmol) were reacted to give 122a as a yellow solid (1500 mg, 75%). LCMS: [M+H]+ 337 Example 122b (2R, 5S)-tert-Butyl 4-(6-Aminopyridinyl)-2,5- dimethylpiperazinecarboxylate 122b Boc N N N NH2 122b 10 Following the procedures as described for nd 101h, reaction of 122a (1.5 g, 4.46 mmol) afforded 122b as a yellow solid (1130 mg, 83%). LCMS: [M+H]+ 307 Example 122c (2R, 5S)-tert Butyl 4-(6-(5-Bromomethyloxo-1,2- dihydropyridinylamino)pyridinyl)-2,5-dimethylpiperazinecarboxylate 122c Boc N N N NH O N Br 122c 15 Following the procedures as described for compound 101i, reaction of 122b (766 mg, 2.50 mmol) and 3,5-dibromomethylpyridin-2(1H)-one (668 mg, 2.50mmol) afforded 122c as a yellow solid (978 mg, 79%). LCMS: [M+H]+ 492 Example 122d (2R, rt-Butyl 4-(6-(5-Bromomethyloxo-1,2- dihydropyridinylamino)pyridinyl)-2,5-dimethylpiperazinecarboxylate 122d 147 HN N N NH O N Br 122d Following the procedures as described for compound 101j, on of 122c (978 mg, 1.99 mmol) gave 122d as a yellow solid (700 mg, 90%). LCMS: [M+H]+ 392 Example 122e 5-Bromo(5-((2S, 5R)-2,5-dimethyl(oxetan 5 yl)piperazinyl)pyridinylamino)methylpyridin-2(1H)-one 122e Following the procedures as described for compound 101k, reaction of 122d (700 mg, 1.79 mmol), afforded 122e as a yellow solid (723 mg, 91%). LCMS: [M+H]+ 448 Example 122f (4-(5-(5-((2S, 5R)-2,5-Dimethyl(oxetanyl)piperazin 10 idinylamino)methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl Acetate 122f O N N N NH AcO O N N N O N 122f Following the procedures as described for compound 113j, reaction of 122e (723 mg, 1.62 mmol) and 3-(acetoxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 15 2(1H)-yl)pyridinylboronic acid 113i (613 mg, 1.62 mmol) afforded 122f as a yellow solid (464 mg, 41%). LCMS: [M+H]+ 707 148 Example 122 2-(4-(5-(5-((2S,5R)-2,5-Dimethyl(oxetanyl)piperazin idinylamino)methyloxo-1,6-dihydropyridinyl)(hydroxymethyl)pyridin yl)-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 122 Following the procedures as described for compound 113, hydrolysis of 122f (464 mg, 5 0.66 mmol) with lithium hydroxide afforded 122 as a white solid (83 mg, 20%). LCMS: [M+H]+ 665. 1H NMR (500 MHz, CDCl3) δ 8.69 (d, J=2.5, 1H), 8.51 (d, J=5.0,1H), 8.03 (d, J=2.5, 1H), 7.88 (s, 1H), 7.86 (d, J=2.5, 1H), 7.38 (d, J=5.0, 2H), 6.90 (s, 1H), 6.82 (d, J=9.0, 1H), 5.07 (s, 1H), .72 (m, 2H), 4.68-4.61 (m, 3H), 4.52 (s, 1H), 4.33 (s, 1H), 4.17-4.11 (m, 2H), 3.88 (s, 1H), 3.76 (s, 1H), 3.73 (s, 3H), 3.19 (s, 1H), 2.93-2.90 (m, 1H), 2.73 (s, 2H), 10 2.63-2.57 (m, 4H), 2.48 (s, 1H), 1.99-1.90 (m, 3H), 1.80 (s, 2H), 0.91 (t, J=5.5, 6H) e 123a (2-Bromoethoxy)(tert-butyl)dimethylsilane 123a To a solution of 2-bromoethanol (5.0 g, 40.3 mmol) in DCM (20 mL) was added tertbutyldimethylsilyl chloride (9.1 g, 60.5 mmol) followed by the additions of triethylamine 15 (8.14 g, 80.6 mmol) and 4-dimethylaminopyridine (49.2 mg, 0.4 mmol). The mixture was stirred at room temperature for 15 h and trated in vacuo. The e was partitioned between 1N HCl and ethyl acetate. The aqueous portion was extracted with ethyl acetate. The combined organic portion was washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo to afford yellow oil, which was purified by column chromatography 20 eluting with PE:EA (50:1) to afford 123a as colorless oil (6.0 g, 62.4 %). LCMS: (M+H)+ 241.
Example 123b 5-Bromo(5-(4-(2-(tertbutyldimethylsilyloxy )ethyl)piperazinyl)pyridineylamino)methylpyridin-2(1H)-one 123b 25 149 To a suspension of 123a (231 mg, 0.96 mmol) in MeCN (40 mL) at 70 oC was added 5-bromomethyl(5-(piperazinyl)pyridinylamino)pyridin-2(1H)-one 101j (350 mg, 0.96 mmol). The reaction mixture was stirred for 3 days. It was then filtered and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica-gel 5 column tography eluting with dichloromethane/methanol (30:1) to afford 123b as yellow solid (452 mg, 90 %). MS: [M+H]+ 524.7. e 123c 5-Bromo(5-(4-(2-hydroxyethyl)piperazinyl)pyridin ylamino)methylpyridin-2(1H)-one 123c HO N N N NH O N Br 123c 10 To a suspension of 123b (300 mg, 0.57 mmol) at room temperature in MeOH (20 mL) was added L(-)-camphorsulfonic acid (199 mg, 0.86 mmol). The reaction e was stirred overnight. Water (20 mL) was added and the mixture was extracted with ethyl acetate (50 mL X 2). The combined organic layer was dried over anhydrous sodium sulfate and concentrated to afford 123c (325 mg, 95%) as a yellow solid. MS: [M+H]+ 408.7. 15 Example 123d (4-(5-(5-(4-(2-Hydroxyethyl)piperazinyl)pyridin ylamino)methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl acetate 123d A sealed tube was charged with 123c (200 mg, 0.49 mmol), 3-(acetoxymethyl)(1- 20 oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinylboronic acid (113i) (188 mg, 0.49 mmol), Pd(dppf)Cl2 (40 mg, 0.049 mmol), K3PO4 (208 mg, 0.98 mmol), 150 NaOAc (133 mg, 0.98 mmol), H2O (3 mL), and MeCN (50 mL). The e was heated at 110o for 3 h. The solvent was evaporated in vacuo and the residue was purified by silica gel chromatography eluting with 30:1 DCM/MeOH to 123d (187 mg, 57 %). MS: [M+H]+ 667.7.
Example 123 2-(4-(5-(5-(4-(2-Hydroxyethyl)piperazinyl)pyridin 5 ylamino)methyloxo-1,6-dihydropyridinyl)(hydroxymethyl)pyridinyl)- 3,4,6,7,8,9-hexahydro-pyrazino[1,2-a]indol-1(2H)-one 123 A mixture of 123d (187 mg, 0.28 mmol) and LiOH (235 mg, 5.6 mmol) in iPrOH/THF (1:1, 3.5 mL) and H2O (0.5 mL) was d at 35 oC for 0.5 h. It was then evaporated in vacuo and the residue was extracted with EtOAc (5 mL X 2). The combined 10 EtOAc extract was concentrated under reduced pressure and the e was purified by reverse-phase prep-HPLC to afford 123 (40 mg, 31 %) as a yellow solid. MS: [M+H]+ 625.4. 1H NMR (500 MHz, CDCl δ 8.63 (d, J = 2.5, 1H), 8.49 (d, J = 5.0, 1H), 7.92 (d, J = 2.5, 3) 1H), 7.82 (d, J = 2.0, 1H), 7.78 (s, 1H), 7.36 (d, J = 5.5, 1H), 7.27-7.25 (m, 1H), 6.89 (s, 1H), 6.81 (d, J = 9.5, 1H), 5.04-5.02 (m, 1H), 4.62 (d, J = 10, 1H), 4.50-4.47 (m, 1H), 4.34 -4.29 15 (m, 1H), 4.12 -4.09 (m, 2H), 3.89-3.85 (m, 1H), 3.71-3.67 (m, 5H), 3.15-3.12 (m, 4H), 2.74- 2.54 (m, 10H), 1.92-1.87 (m, 2H), 1.79-1.78 (m, 3H) Example 124a 4-Chloro{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca- 1(9),2(7),3- trienyl}pyridinecarbaldehyde 124a To a suspension of 2-bromochloronicotinaldehyde 103a (641 mg, 2.9 mmol) and 20 8-thia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienone 191d (400 mg, 1.94 mmol) in dioxane (20 mL) was added K2CO3 (536 mg, 3.88 mmol), CuI (369 mg, 1.94 mmol), and 4,7-dimethoxy-1,10-phenanthroline (471 mg, 1.96 mmol). After bubbling nitrogen through the resulting solution for 30 min, the e was d at 80 ºC for 16 h. It was allowed to cool to room temperature and added into H2O (100 mL). The aqueous layer was separated 25 and extracted with ethyl acetate (2 × 200 mL). The combined organic layer was washed with brine (100 mL) and dried over sodium sulfate. The drying agent was d by filtration and the filtrate was concentrated under reduced pressure. The residue was purified on flash column eluting with PE:EA (5:1) to afford 124a (230 mg, 34%). LCMS: [M+H]+ 346 Example 124b ethyl(5-(4-(oxetanyl)piperazinyl)pyridin 30 ylamino)oxo-1,6-dihydropyridinyl]{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca- 1(9),2(7),3- 5-yl}pyridinecarbaldehyde 124b A round bottom flask was charged with 124a, 1-methyl(5-(4-(oxetan yl)piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin- 2(1H)-one 101l (271 mg, 0.58 mmol), PdCl2(dppf) (50 mg, 0.06 mmol), K3PO4.3H2O (323 151 mg, 1.16 mmol), THF (15 mL), and H2O (5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 70 oC for 2 h. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified on flash column chromatography eluting with 1:3 petroleum/ethyl acetate to afford 124b as a yellow solid (200 mg, 53%). LCMS: [M+H]+ 651 5 Example 124 oxymethyl[1-methyl(5-(4-(oxetanyl)piperazin- 1-yl)pyridineylamino)oxo-1,6-dihydropyridinyl]{6-oxothia-4,5- diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridine 124 A mixture of 4-[1-methyl(5-(4-(oxetanyl)piperazinyl)pyridinylamino) oxo-1,6-dihydropyridinyl]{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3- 10 trienyl}pyridinecarbaldehyde 124b (200 mg, 0.31 mmol), NaBH4 (35 mg, 0.92 mmol) and CH3OH (10 mL) was stirred at 25oC for 1 h. The mixture was then extracted with CH2Cl2 (10 mL X 2). The combined CH2Cl2 extract was concentrated under reduced pressure. The residue was purified with reverse-phase prep-HPLC to afford 124 (100 mg, 50%) as a yellow solid. LCMS: [M+H]+ 653. 1H NMR (500 MHz, DMSO) δ 8.64 (d, J=2.0 Hz, 1 H), 8.57 (d, 15 J=5.0 Hz, 1 H), 8.46-8.48 (m, 2 H), 7.88 (d, J=3.0 Hz, 1 H), 7.54 (d, J=5.0 Hz, 1 H), 7.48 (d, J=2.5 Hz, 1 H), 7.37-7.39 (m, 1 H), 7.24 (d, J=9.0 Hz, 1 H), 4.85-4.87 (m, 1 H), 4.55-4.57 (m, 2 H), 4.45-4.47 (m, 2 H), .39 (m, 2 H), 3.60 (s, 3 H), 3.42-3.45 (m, 1 H), .08 (m, 4 H), 2.95 (s, 2 H), 2.87 (s, 2 H), 2.38-2.40 (m,4 H), 1.87-1.89 (m, 4 H).
Example 125a ro-1H-pyrazolyl)methanol 125a O2N OH N N H 125a 20 A 3-L three-neck round-bottomed flask ed with a mechanical stirrer, addition funnel and nitrogen inlet was purged with nitrogen and d with 3-nitropyrazole carboxylic acid (28.0 g, 178 mmol) and THF (420 mL) and cooled to –5 °C using an ice/acetone bath. Borane-THF complex solution (1.0 M, 535 mL, 535 mmol) was added at a 25 rate that maintained the internal reaction temperature below 5 °C. After the addition was te the cooling bath was removed and the reaction was stirred at room temperature for 18 h. After this time the reaction was cooled to –5 °C using an ice/acetone bath, water (70 mL) and 4N hydrochloric acid (70 mL) was added and the reaction was stirred at reflux for 1 h in order to destroy the borane x with pyrazole. The reaction was cooled to room 30 temperature and concentrated under reduced pressure to a volume of approximately 30 mL.
Ethyl acetate (175 mL) was added and the mixture stirred for 15 min. The aqueous layer was 152 separated and extracted with ethyl acetate (4 × 200 mL). The combined organic layers were washed with saturated aqueous sodium bicarbonate (2 × 50 mL), brine (50 mL) and dried over sodium sulfate, the drying agent was removed by tion, and the filtrate concentrated under reduced pressure to afford 125a in a 94% yield (24.0 g) as a light yellow solid: 1H 5 NMR (300 MHz, DMSO-d6) δ 13.90 (br s, 1H), 6.87 (s, 1H), 5.58 (t, 1H, J = 5.4 Hz), 4.53(d, 2H, J = 5.1 Hz); MS (ESI+) m/z 144.0 (M+H) Example 125b (1-(2-Bromoethyl)nitro-1H-pyrazolyl)methanol 125b O2N OH N N Br 125b A 1-L three-necked round-bottomed flask equipped with a mechanical stirrer and 10 regulator was purged with nitrogen and charged with 125a (25.0 g, 175 mmol), DMF (250 mL), and cesium carbonate (70.0 g, 215 mmol) was heated at 104 °C for 5 min. The reaction mixture was then cooled to 0 °C using an ice/acetone bath and dibromoethane (329 g, 1.75 mol) was added portionwise (no rm). The reaction was stirred at 0 °C for 1 then at room temperature for 4 h. After this time a solution of KH2PO4 (40 g) in water (400 mL) 15 was added slowly. The reaction mixture stirred at room temperature for 30 min. Ethyl acetate (450 mL) was added and the aqueous layer was separated and extracted with ethyl acetate (2 × 100 mL). The ed organic layers were washed with water (200 mL), brine (200 mL), dried over sodium sulfate, and the drying agent was removed by filtration. The filtrate was trated under reduced re to afford an 86% yield (37.5 g) of crude 20 125b as an orange oil: 1H NMR (300 MHz, CDCl3) δ 6.85 (s, 1H), 4.82 (d, 2H, J = 5.4 Hz), 4.66 (t, 2H, J = 6.3 Hz), 3.83 (t, 2H, J = 6.3 Hz); MS (ESI+) m/z 249.9 (M+H).
Example 125c 1-(2-Bromoethyl)(bromomethyl)nitro-1H-pyrazole 125c O2N Br N N Br 125c A 500-mL three-necked round-bottomed flask equipped with a magnetic stirrer, 25 nitrogen inlet and reflux condenser was purged with nitrogen and charged with 125b (37.0 g, 148 mmol) and chloroform (160 mL). The reaction was cooled to –5 °C using an ice/acetone bath and phosphorous mide (40.0 g, 148 mmol) was added portionwise. The cooling bath was removed and the reaction stirred at reflux for 2 h. After this time, the reaction was 153 cooled to –5 °C and saturated aqueous sodium bicarbonate (250 mL) was added until a pH of 8.5 was reached. The mixture was extracted with ethyl acetate (3 × 150 mL) and the combined organic layers were washed with saturated aqueous sodium carbonate (2 × 50 mL), brine (75 mL), dried over sodium sulfate and the drying agent was removed by filtration. The 5 filtrate was concentrated under reduced pressure to afford a yellow e that was dissolved with gentle heating in methylene de (60 mL). Hexanes (approximately 20 mL) was added and the solution became cloudy. The mixture was heated until a solid precipitate formed, methylene chloride (9 mL) was added and the solution became clear. The solution was left to cool to room temperature and after 4 h the resulting crystals were collected by 10 vacuum filtration. The filter cake was washed with a ice cold 1:2 mixture of methylene chloride:hexanes (2 × 20 mL) to afford 1-(2-bromoethyl)(bromomethyl)nitro-1H- le (19.7 g). The combined filtrates were evaporated and the procedure was med again to afford an additional 9.70 g of 1-(2-bromoethyl)(bromo-methyl)nitro-1H- pyrazole. The solids were combined and dried under high vacuum for 18 h to afford a 57% 15 yield (26.0 g) of 125c as white ls: mp 95–97 °C; 1H NMR (300 MHz, CDCl3) δ 6.93 (s, 1H), 4.63 (t, 2H, J = 6.0 Hz), 4.54 (s, 2H), 3.86 (t, 2H, J = 6.0 Hz).
Example 125d 2-nitro-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine 125d A sealed tube ed with a magnetic stirrer was charged with 125c (4 g, 12.9 20 mmol) 0.5M ammonia solution in dioxane (200 mL). The resulting mixture was carefully heated to 50 °C overnight. After this time, the reaction mixture was concentrated under reduced pressure, and to the residue was added H2O (50 mL) and EtOAc (50 mL). The aqueous layer was separated and extracted with EtOAc (2 × 50 mL). The combined organic extracts were washed with brine (100 mL) and dried over sodium sulfate. The resulting 25 solution was trated under reduced pressure to afford a 100% yield (2.1 g) of crude 125d.
Example 125e 1-(2-nitro-6,7-dihydropyrazolo[1,5-a]pyrazin-5(4H)-yl)ethanone 125e 154 A 200 mL round bottom flask was charged with 125d (2.1g, 12.9 mmol), triethylamine (5.5 mL, 38.7 mmol), acetyl chloride (1.1 mL, 15.5 mmol) and CH2Cl2 (100 mL). The mixture stirred at room temperature over night. After this time, the reaction 5 mixture was concentrated under reduced pressure, and to the residue was added H2O (50 mL) and EtOAc (50 mL). The aqueous layer was separated and extracted with EtOAc (2 × 50 mL).
The combined organic extracts were washed with brine (100 mL). The combined aqueous extracts were back ted with 9:1 CH2Cl2:MeOH (2 x 50 mL). The combined organics were dried over sodium sulfate. The resulting e was purified by column 10 chromatography g with a gradient of CH2Cl2 – 9:1 CH2Cl2: MeOH to afford a 84% yield (2.3 g) of 125e.
Example 125f 1-(2-amino-6,7-dihydropyrazolo[1,5-a]pyrazin-5(4H)- anone 125f 15 A 500-mL Parr hydrogenation bottle was charged with 125e (2.3 g, 10.9 mmol), 10% palladium on carbon (50% wet, 570 mg dry weight) and ethanol (100 mL). The bottle was evacuated, charged with hydrogen gas to a pressure of 50 psi and shaken for 2 h on a Parr hydrogenation apparatus. The catalyst was removed by filtration h a pad of CELITE® 521 g with 1:1 CH2Cl2:MeOH (500mL). The resulting solution was concentrated 20 under reduced pressure to afford a 95% yield (1.9 g) of crude 125f.
Example 125g 3-(5-acetyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin ylamino)bromomethylpyridin-2(1H)-one 125g 155 A sealed tube was equipped with a magnetic stirrer and charged with 125f (860 mg, 4.8 mmol), 3,5-dibromomethylpyridin-2(1H)-one (1.8 g, 6.7 mmol), and cesium carbonate (3.4 g, 10.5 mmol) in 1,4-dioxane (67 mL). After bubbling nitrogen through the solution for 30 min, Xantphos (330 mg, 0.6 mmol) and tris(dibenzylideneacetone) dipalladium(0) (300 5 mg, 0.3 mmol) were added, and the reaction mixture was heated to 100 °C for 16 h. After this time, H2O (50 mL) and EtOAc (50 mL) were added. The aqueous layer was separated and extracted with EtOAc (2 × 50 mL). The combined c extracts were washed with brine (100 mL) and dried over sodium sulfate. The resulting residue was purified by column tography eluting with a gradient of CH2Cl2 – 60:35:5 CH2Cl2:Et2O:MeOH to afford a 10 41% yield of 125g (720 mg).
Experiment 125h omethyl(4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinylamino)pyridin-2(1H)-one 125h A 50 mL round bottom flask with a magnetic stirrer and reflux condenser was charged 15 with 125g (250 mg, 0.7 mmol), aqueous NaOH (5N, 6 mL), ethanol (6 mL). The mixture stirred at reflux for 30 min. After this time, ethyl acetate (5 mL) and water (5 mL) were added. The separated s layer was extracted with ethyl acetate (2 x 5 mL). The combined organics were washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under d pressure to afford a 91% yield (200 mg) of crude 125h. 20 Example 125i 5-Bromomethyl(5-(oxetanyl)-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)pyridin-2(1H)-one 125i O N N N NH O 125i N Br Compound 125i was synthesized using the same procedure as 101k, where 5-bromo- 1-methyl(4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinylamino)pyridin-2(1H)-one (125h) 25 (250 mg, 0.78 mmol), and oxetanone (600 mg, 8.3 mmol) in methanol (8 mL) were mixed.
Sodium cyanoborohydride (148 mg, 3 mmol) and zinc chloride (165 mg, 1.5 mmol) in 156 methanol (8 mL) was added, and the on was heated at 48 oC for 12 hours. Work-up and flash column chromatography (silica, 60:35:5 methylene chloride/diethyl ether/methanol) afford a 34% yield (100 mg) of 5-bromomethyl(5-(oxetanyl)-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)pyridin-2(1H)-one (125i) as a light green solid: 5 MS (ESI+) m/z 382.1 (M+H).
Example 125j (4-(1-Methyl(5-(oxetanyl)-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)oxo-1,6-dihydropyridinyl)(1-oxo- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl Acetate 125j O N N N NH AcO O N N N O N 125j 10 Following the procedures as described for compound 113j, 3-(acetoxymethyl)(1- oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinylboronic acid 113i (200 mg, 0.52 mmol) and 125i (198 mg, 0.52 mmol) were reacted to give 125j as a yellow solid (200 mg, 60%). LCMS: [M+H]+ 639 Example 125 2-(3-(Hydroxymethyl)(1-methyl(5-(oxetanyl)-4,5,6,7- 15 tetrahydropyrazolo[1,5-a]pyrazinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 125 Following the procedures as described in Example 123, 125j (200 mg 0.31 mmol) was hydrolyzed by lithium hydroxide to give 125 as a white solid (116 mg, 62%). LCMS: [M+H]+ 597. 1H NMR (500 MHz, CDCl3) δ 8.48 (d, J=5.0, 1H), 7.95 (d, J=2.0, 1H), 7.69 (d, 20 J=2.0, 1H), 7.43 (s, 1H), 7.34 (d, J=5.5, 1H), 6.89 (s, 1H), 5.73 (s, 1H), 5.02 (t, J=6.5, 1H), 4.75 (t, J=6.5, 2H), 4.67 (t, J=6.5, 2H), 4.61-4.63 (m, 1H), 4.50 (s, 1H), 4.31-4.35 (m, 1H), .16 (m, 4H), 3.86-3.88 (m, 1H), 3.74-3.79 (m, 1H), 3.70 (s, 3H), 3.56 (d, J=4.5, 2H), 2.82 (t, J=4.5, 2H), .62 (m, 4H), 1.88-1.92 (m, 2H), 1.78-1.82 (m, 2H) Example 126a 3-Bromo(1-oxo-6,7,8,9-tetrahydropyrazino[1,2-a]indol- 25 2(1H)-yl)isonicotinaldehyde 126a 157 A 100-mL single-neck round-bottomed flask equipped with a ic stirrer and reflux ser was charged with 1,4-dioxane (15 mL), 3,5-dibromoisonicotinaldehyde (604 mg, 2.28 mmol), 6,7,8,9-tetrahydropyrazino[1,2-a]indol-1(2H)-one (142 mg, 0.76 mmol) and 5 cesium carbonate (485 mg, 1.5 mmol). CuI (143 mg, 0.76 mmol) and 4,7-dimethoxy-1,10- phenanthroline (127 mg, 0.52 mmol) were added, and the on mixture was heated at 100 ºC for 5 h. After this time, the reaction was cooled to room temperature. It was then filtered and the filtrate was concentrated under d pressure. The residue was purified on flash column eluting with EtOAC/PE (1:2) to afford 126a (100 mg, 35%) as a yellow solid. MS: 10 [M+H]+ 372.
Example 126b 3-(1-Methyl(5-(4-(oxetanyl)piperazinyl)pyridin ylamino)oxo-1,6-dihydropyridinyl)(1-oxo-6,7,8,9-tetrahydropyrazino[1,2-a]indol- 2(1H)-yl)isonicotinaldehyde 126b 15 A sealed tube was charged with 126a (100 mg, 0.27 mmol)，1-methyl(5-(4- (oxetanyl)piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolan yl)pyridin-2(1H)-one 101l (125 mg, 0.27 mmol), PdCl2(dppf) (18 mg, 0.02 mmol), K3PO4 (30 mg), and NaOAc (20 mg) in CH3CN (5 mL) and H2O (1 mL). The system was evacuated and refilled with N2. The reaction mixture was heated at 100 ºC for 2 h, and then cooled to 20 room temperature. It was then filtered and the te was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography eluting with 10:1 of DCM/MeOH to afford 126b (80 mg, 48%) as a yellow solid. MS: [M+H]+ 633.
Example 126 2-(4-(Hydroxymethyl)(1-methyl(5-(4-(oxetan yl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)-6,7,8,9- 25 tetrahydropyrazino[1,2-a]indol-1(2H)-one 126 158 To a suspension of 126b (76 mg, 0.12 mmol) at 0 oC in methanol (4 mL) was added sodium borohydride (20 mg, 0.7 mmol) and the mixture was stirred for 30 minutes. Then the reaction mixture was quenched with water (1.0 mL) and concentrated. The e was purified by reverse-phase prep-HPLC to afford 126 (28 mg, 37 %). LCMS: [M+H]+ 635. 1H 5 NMR (500 MHz, DMSO) δ 8.61 (d, J=2.5, 1H), 8.59 (s, 1H), 8.50 (s, 1H), 8.43 (s, 1H), 7.86 (d, J=3.0, 1H), 7.38-7.36 (m, 2H), .22 (m, 2H), 6.82 -6.78 (m, 2H), 5.18-5.11 (m, 1H), 4.55 (t, J=6.0, 2H), 4.45 (t, J=6.0, 2H), 4.41-4.29 (m, 2H), 3.60 (s, 3H), 3.44-3.42 (m, 1H), 3.06 (t, J=4.5, 4H), 2.75-2.73 (m, 2H), 2.62-2.60(m, 2H), 2.38 (t, J=4.5, 4H), 1.86-1.75 (m, 4H). 10 Example 127a (4-(1-Methyloxo(5-(piperazinyl)pyridinylamino)- 1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- yl)pyridinyl)methyl Acetate 127a A 100-mL single-neck round-bottomed flask ed with magnetic stirrer and 15 reflux condenser was charged with 5-bromomethyl(5-(piperazinyl)pyridin ylamino)pyridin-2(1H)-one 101j (200 mg, 0.55 mmol), 3-(acetoxymethyl)(1-oxo- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinylboronic acid 113i (210 mg, 0.55 mmol), Pd(dppf)Cl2 (45 mg, 0.055 mmol), K3PO4 (284 mg, 1.65 mmol), and tetrahydrofuran (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at 20 reflux for 5 h. It was then cooled to room ature and filtered. The filtrate was concentrated under reduced re and the resulting residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (33:1) to afford 127a as a brown solid (200 mg, 58.3 %).MS: [M+H]+ 623.7.
Example 127 2-(3-(Hydroxymethyl)(1-methyloxo(5-(piperazin 25 yl)pyridinylamino)-1,6-dihydropyridinyl)pyridinyl)-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-1(2H)-one 127 A mixture of 127a (190 mg, 0.31 mmol) and LiOH (571 mg, 13.6 mmol) in iPrOH/THF (1:1, 3.5 mL) and H oC for 0.5 h. It was then 2O (0.5 mL) was stirred at 35 159 evaporated in vacuo and the residue was extracted with EtOAc (5 mL X 2). The combined EtOAc extract was concentrated under reduced pressure and the residue was ed by reverse-phase prep-HPLC to afford 127 (50 mg, 26.9 %). MS: [M+H]+ 581.3. 1H NMR (500 MHz, CDCl3) δ 8.63 (d, J = 2.0, 1H), 8.49 (d, J = 5.0, 1H), 7.91 (d, J = 3.5, 1H), 7.82 (d, J = 5 2.0, 1H), 7.77 (s, 1H), 7.37 (d, J = 5.0,1H), 7.20-7.25 (m,1H), 6.89 (s, 1H), 6.81 (d, J = 9.0,1H), 5.04-5.02 (m, 1H), 4.64-4.61 (m, 1H), 4.50 (d, J = 5.0, 1H), 4.34-4.31 (m, 1H), 4.18- 4.08 (m, 2H), 3.89-3.86 (m, 1H), 3.71 (s, 3H), .06 (m, 8H), 2.62-2.56 (m, 4H), 1.92- 1.88 (m, 2H), 1.81-1.78 (m, 3H) Example 128a 5-Cyclopropylnitro-4,5,6,7-tetrahydropyrazolo[1,5- 10 a]pyrazine 128a N N N NO2 128a A mixture of 1-(2-bromoethyl)(bromomethyl)nitro-1H-pyrazole 113c (4 g, 12.9 mmol) and cyclopropanamine (7.35 g, 129 mmol) in THF (40 mL) was stirred at 30°C overnight. After the completion of the on, the mixture was filtered and the solid was 15 washed with THF (100 mL). The filtrate was concentrated under reduced re to give 128a (2.68 g, 99%). MS: [M+H]+ 209.
Example 128b 5-Cyclopropyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin amine 128b 20 A e of 128a (2.68 g, 12.9 mmol), Fe (3.6 g, 64.4 mmol) and NH4Cl (4.1 g, 77.4 mmol) in ethanol (30 mL) and water (5 mL) was heated at reflux for 2 h. After the completion of the reaction, the mixture was filtered and the solid was washed with ethanol (150 mL). The filtrate was ated in vacuo and the residue was extracted with methanol /methylene chloride (1/7). The combined extracts were dried over Na2SO4 and evaporated. 25 The residue was purified on reverse phase Combi-flash to give 128b (1.8 g, 75%). MS: [M+H]+ 179.
Example 128c 5-Bromo(5-cyclopropyl-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinylamino)methylpyridin-2(1H)-one 128c 160 A mixture of 128b (1.39 g, 7.8 mmol), XantPhos (450 mg, 0.78 mmol), Pd2dba3 (476 mg, 0.52 mmol), 3,5-dibromomethylpyridin-2(1H)-one (1.72 g, 6.5 mmol) and Cs2CO3 (6.3 mg. 19.5 mmol) in 1,4-dioxane (30 mL) was heated at reflux for 1 h. After the 5 completion of the on the mixture was filtered off and the solid was washed with methanol (60 mL). The filtrate was evaporated in vacuo and the residue was purified on reverse phase Combi-flash to give 128c (0.84 g, 30%). MS: [M+H]+ 364.
Example 128d (4-(5-(5-Cyclopropyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin- 2-ylamino)methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydro- 10 pyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl e 128d N N N NH AcO O N N N O N 128d Following the procedures as described in Example 113j, reaction of 128c (230 mg, 0.6 mmol) and 3-(acetoxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- yl)pyridinylboronic acid 113i (218 mg, 0.6 mmol) afforded 128d as a yellow solid (331 15 mg, 89%). LCMS: [M+H]+ 623 Example 128 2-(4-(5-(5-Cyclopropyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin o)methyloxo-1,6-dihydropyridinyl)(hydroxymethyl)pyridinyl)- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 128 Following the procedures as described in Example 113, 128d (331 mg, 0.53 mmol) 20 was hydrolyzed with m hydroxide afforded 128 as a white solid (54 mg, 20%). LCMS: [M+H]+ 581. 1H NMR (500 MHz, CDCl3) δ 8.48 (d, J=5.0, 1H), 7.93 (d, J=2.0, 1H), 7.72 (d, J=2.0, 1H), 7.40 (s, 1H), 7.34 (d, J=5.0, 1H), 6.90 (s, 1H), 5.70 (s,1H), 5.03-5.02 (m, 1H), 4.64-4.62 (m, 1H), 4.52 (s, 1H), 4.32 (s, 1H), 4.16-4.03 (m, 4H), 3.89-3.87 (m, 1H), 3.80 (s, 161 2H), 3.70 (s, 3H), 3.12-3.10 (m, 2H), 2.61-2.57 (m, 4H), 1.90 (d, J=5.5, 3H), 1.79 (s, 2H), 0.56 (d, J=6.0, 2H), 0.53 (s, 2H) Example 129a 2-Nitro-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazine 129a 5 A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and reflux condenser was charged with 1-(2-bromoethyl)(bromomethyl)nitro-1H-pyrazole 113c (3.00 g, 9.59 mmol) and 4M aqueous hydrobromic acid (120 mL), and the resulting mixture was heated at reflux for 24 h. After this time, the reaction mixture was concentrated under reduced pressure to approximately 6 mL volume, and the residue was stirred in 2M 10 aqueous sodium hydroxide (40 mL) for 2 h. After this time methylene chloride was added (40 mL) and the mixture was stirred for 15 min. The aqueous layer was separated and extracted with ene chloride (2 × 50 mL). The combined organic extracts were washed with brine (100 mL) and dried over sodium e. The drying agent was removed by filtration and the filtrate concentrated under reduced pressure to afford a 62% yield (1.01 g) 15 of 129a as a white solid: mp 110–112 °C; 1H NMR (300 MHz, CDCl3) δ 6.68 (s, 1H), 4.87 (s, 2H), 4.28 (t, 2H, J = 5.4 Hz), 4.20 (t, 2H, J = 5.1 Hz); MS (ESI+) m/z 170.0 (M+H).
Example 129b 6,7-Dihydro-4H-pyrazolo[5,1-c][1,4]oxazinamine 129b A 500-mL Parr hydrogenation bottle was purged with en and charged with 129a 20 (1.01 g, 5.92 mmol), 10% palladium on carbon (50% wet, 125 mg dry weight) and l (50 mL). The bottle was evacuated, charged with hydrogen gas to a pressure of 25 psi and shaken for 2 h on a Parr hydrogenation apparatus. The hydrogen was then evacuated and nitrogen charged to the bottle. The catalyst was removed by filtration through a pad of CELITE® 521 and the filtrate concentrated under reduced re. The ing residue 25 was purified by column tography using 400 cc of silica gel and eluting with 3% ol in methylene chloride. The fractions containing 129b were collected to afford, after concentrating under reduced pressure, a 73% yield (601 mg) of 129b as a yellow solid: mp 74–76°C 1H NMR (300 MHz, CDCl3 δ 5.37 (s, 1H), 4.72 (s, 2H), 4.07 (t, 2H, J = 5.1 Hz), 3.98 (t, 2H, J = 5.1 Hz), 3.57 (br s, 2H); MS (ESI+) m/z 140.4 (M+H). 162 Example 129c 5-Bromo(6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin ylamino)methylpyridin-2(1H)-one 129c O N N NH O 129c N Br CH3 A 50-mL three-neck round-bottomed flask equipped with a magnetic stirrer, reflux 5 condenser and en inlet was charged with 1,4-dioxane (20 mL), 129b (600 mg, 4.31 mmol), bromomethyl pyridine–2(1H)-one (1.44 g, 5.40 mmol) and cesium carbonate (3.08 g, 9.48 mmol). After bubbling nitrogen through the resulting solution for 30 min, Xantphos (300 mg, 0.52 mmol) and tris(dibenzylideneacetone)dipalladium(0) (320 mg, 0.35 mmol) were added, and the reaction mixture was heated at reflux for 2 h. After this time the 10 reaction was cooled to room temperature, partitioned between ethyl acetate (75 mL) and water (75 mL) and filtered. The aqueous layer was separated and extracted with ethyl acetate (2 × 25 mL). The organic layers were combined and washed with brine (50 mL) and dried over sodium sulfate. The drying agent was removed by tion and the filtrate concentrated under reduced pressure. The resulting residue was purified by column tography using 15 500 cc of silica gel and eluting with 1% methanol in methylene de. The fractions containing 129c were collected to afford, after trating under reduced pressure, a 31% yield (433 mg) of 129c as a green solid: mp 195–197 °C; 1H NMR (300 MHz, CDCl3) δ 7.92 (d, 1H, J = 2.4 Hz), 7.44 (s, 1H), 6.90 (d, 1H, J = 2.4 Hz), 5.65 (s, 1H), 4.80 (s, 2H), 4.13 (s, 2H), 3.61 (s, 5H); MS (ESI+) m/z 324.9 (M+H). 20 e 129d (4-(5-(6,7-Dihydro-4H-pyrazolo[5,1-c][1,4]oxazinylamino)- 1-methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)pyridinyl)methyl Acetate 129d Following the procedures as described in Example 113j, reaction of 3- 25 (acetoxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridin 163 ylboronic acid 113i (200 mg, 0.52 mmol) and 129c (170 mg, 0.52 mmol) gave 129d as a yellow solid (185mg, 61%). LCMS: [M+H]+ 584 e 129 2-(4-(5-(6,7-Dihydro-4H-pyrazolo[5,1-c][1,4]oxazinylamino) methyloxo-1,6-dihydropyridinyl)(hydroxymethyl)pyridinyl)-3,4,6,7,8,9- 5 hexahydropyrazino[1,2-a]indol-1(2H)-one 129 Following the procedures as described in Example 113, 129d (180 mg 0.31 mmol) was hydrolyzed with lithium hydroxide to give 129 as a white solid (100 mg, 62%). LCMS: [M+H]+ 542. 1H NMR (500 MHz, CDCl3) δ 8.48 (d, J=5.0, 1H), 7.98 (d, J=2.0, 1H), 7.71 (d, J=2.0, 1H), 7.46 (s, 1H), 7.35 (d, J=5.0, 1H), 6.89 (s, 1H), 5.72 (s, 1H), 5.03 (d, J=6.5, 1H), 10 4.79 (s, 2H), 4.61-4.64 (m, 1H), 4.50 (s, 1H), 4.31-4.35 (m, 1H), .16 (m, 6H), 3.86 (s, 1H), 3.71 (s, 3H), 2.56-2.62 (m, 4H), 1.88-1.92 (m, 2H), 1.80 (m, 2H) Example 130a (3S)-tert-Butyl 3-methyl(6-nitropyridinyl)piperazine carboxylate 130a 15 Following the ures as bed for compound 101g, reaction of 5-bromo nitropyridine (10.5 g, 50 mmol), and (3S)-tert-butylmethylpiperazinecarboxylate (10.0 g, 50 mmol) afforded 130a as a yellow solid (8.05 g, 50%). LCMS: [M+H]+ 323 Example 130b (3S)-tert-butyl(6-aminopyridinyl)methylpiperazine carboxylate 130b 20 Following the procedures as described for compound 101h, hydrogenation of 130a (5.8 g) afforded 130bas a brown solid (4.9 g, 96%). LCMS: [M+H]+ 293 Example 130c (3S)-tert-Butyl(6-(5-bromomethyloxo-1,2- dihydropyridinylamino) pyridineyl)methylpiperazinecarboxylate 130c 164 Following the procedures as described for compound 101i, on of 130b (4.0 g) and 3,5-dibromomethylpyridin-2(1H)-one (5.5 g) afforded 130c as a yellow solid (5.4 g, 83%). LCMS: [M+H]+ 478 5 Example 130d (3S)Bromomethyl(5-(2-methylpiperazinyl)pyridin- ino)pyridine-2(1H)-one 130d HN N N NH O N 130d Br Following the procedures as described for compound 101j, acidic hydrolysis of the Boc group of 130c (3.1 g) afforded 130d as a yellow solid (2.3 g, 95%). LCMS: [M+H]+ 380. 10 Example 130e (3S)Bromomethyl(5-(2-methyl(oxetan erazinyl) pyridine ylamino)pyridin-2(1H)-one 130e Following the procedures as described for compound 101k, reductive amination of 130d (2.35 g) with oxetanone (0.4 mL) afforded 130e as a yellow solid (2.6 g, 98%). 15 LCMS: [M+H]+ 434.
Example 130f (3S)methyl(5-(2-methyl(oxetanyl)piperazin yl)pyridinylamino) (4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 130f 165 O N N N NH O O N B O 130f A 100 mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 130e (1.0 g, 1.0 eq., 2.3 mmol), Pin2B2 (1.46 g, 2.50 eq., 5.75 mmol), Pd2(dba)3 (105 mg, 0.05 eq., 0.125 mmol), X-Phos (93 mg, 0.1 eq., 0.23 mmol), 5 AcOK (676 mg, 3.0 eq., 6.9 mmol), and e (50 mL). After three cycles of vacuum/argon flush, the mixture was heated at 90 ºC for 4 hrs, then cooled to room temperature and filtered. The filtrate was concentrated under d pressure and the resulting residue was washed with 3:1 PE/EA (80 mL) to afford 130f as yellow solid (1.0 g, 90%). MS: [M+H]+ 482. 10 Example 130g (3S)[1-methyl({5-[2-methyl 4-(oxetanyl)piperazin yl]pyridineyl}amino)oxo-1,6-dihydropyridinyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 130g A 50 mL single-neck round-bottomed flask equipped with a ic stirrer and a 15 reflux condenser was charged with 130f (420 mg, 1.0 eq., 0.44 mmol), 4-chloro{4,4- dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridine carbaldehyde 108a (200 mg, 2 eq., 0.88 mmol): , 166 PdCl2(dppf) (36 mg, 0.1 eq., 0.044 mmol), K3PO4 (279 mg, 3 eq., 1.32 mmol), and THF (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at reflux for 5 h. It was then cooled to room temperature and filtered. The te was concentrated under reduced pressure and the ing residue was washed with 3:1 PE/EA (80 mL) to afford 5 130g (90 mg, 31%) as a yellow solid. MS: [M+H]+ 663.
Example 130 (3S)[4-[1-methyl({5-[2-methyl 4-(oxetan yl)piperazinyl]pyridineyl}amino)oxo-1,6-dihydropyridinyl] (hydroxymethyl)pyridinyl]-4,4-dimethyl-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dien- 9-one 130 10 A 50 mL single-neck round-bottomed flask equipped with a magnetic stirrer and was charged with 130g (90 mg, 1 eq., 0.11 mmol), LiOH (7.9 mg, 3 eq., 0.33 mmol), i-PrOH (3 mL), THF (3 mL) and H2O (2 mL). The e was stirred at 30 ºC for 2 h. It was then filtered and concentrated. The residue was purified by reverse-phase prep-HPLC to afford 130 (40 mg, 44%) as a yellow solid. LCMS: [M+H]+ 665.4. 1H NMR (500 MHz, CDCl3) δ 15 8.65 (d, J=2.0, 1H), 8.48 (d, J=5.0, 1H), 7.96 (d, J=2.0, 1H), 7.84-7.83 (m, 2H), 7.36 (d, J=5.0, 1H), 7.31 (dd, J=3.0, 9.0, 1H), 6.84 (s, 1H), 6.81 (d, J=9.0, 1H), .05 (m, 1H), 4.71-4.61 (m, 5H), 4.51-4.29 (m, 2H), 4.16-4.15 (m, 2H), 3.87-3.85 (m, 1H), 3.72 (s, 3H), 3.55-3.45 (m, 2H), 3.06 - 3.08 (m, 2H), 2.59-2.47 ( m, 7H), .17 (m, 1H), 1.27 (s, 6H), 0.98 (d, J=6.5, 3H). 20 Example 131a (S)-(4-(5-(5-(2-ethyl(oxetanyl)piperazinyl)pyridin ylamino)methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl acetate 131a O N N N NH AcO O N N N O N 131a A sealed tube equipped with a magnetic stirrer was charged with (S)bromo(5-(2- 25 ethyl(oxetanyl)piperazinyl)pyridinylamino)methylpyridin-2(1H)-one 161e (269 mg, 0.60 mmol): 167 , 113i (230 mg, 0.60 mmol), Pd(dppf)Cl2 (25 mg, 0.03 mmol), NaOAc (98 mg, 1.2 mmol), K3PO4 (254 mg, 1.2 mmol), and acetonitrile (4 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 1 h. It was then filtered and the 5 filtrate was evaporated in vacuo. The e was purified by silica gel column tography eluting with dichloromethane/methanol (25:1, V/V) to afford 131a (150 mg, 40%) as a brown solid. LCMS: [M+H]+ 707 Example 131 (S)(4-(5-(5-(2-ethyl(oxetanyl)piperazinyl)pyridin ylamino)methyloxo-1,6-dihydropyridinyl)(hydroxymethyl)pyridinyl)- 10 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 131 A mixture of 131a (150 mg, 0.21 mmol) and LiOH (50 mg, 2.1 mmol) in iPrOH/THF (1:1, 4 mL) and H2O (1 mL) was stirred at 30oC for 1 h. The mixture was evaporated in vacuo and the residue was extracted with EtOAc (10 mL X 2). The combined EtOAc extract was concentrated under reduced pressure and the residue was purified by reverse-phase prep- 15 HPLC to afford 131 (26 mg, 25%) as a white solid. LCMS: [M+H]+ 665. 1H NMR (500 MHz, CDCl3) δ 8.64 (d, J=2.0, 1H), 8.50 (d, J=5.0, 1H), 7.93 (d, J=2.5, 1H), 7.83 (d, J=1.5, 2H), 7.38 (d, J=5.0, 1H), 7.27 (d, J=5.0,1H), 6.90 (s,1H), 6.83 (d, J=8.5, 1H), 4.73-4.64 (m, 5H), 4.50 (s, 1H), 4.33-4.31 (m, 1H), 4.20-4.16 (m, 2H), 3.88-3.86 (m, 1H), 3.73 (s, 3H), 3.53- 3.51 (m, 1H), 3.33 (s, 1H), 3.13 (t, J=5.0, 2H), 2.61-2.56 (m, 4H), 2.45 (d, J=4.0, 2H), 2.37 20 , 1.91-1.79 (m, 7H), 1.39-1.40 (m, 1H), 0.83 (t, J=7.0, 3H).
Example 132a 6-Chloro(6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin ylamino)methylpyridazin-3(2H)-one 132a A mixture of hydro-4H-pyrazolo[5,1-c][1,4]oxazinamine 129b (0.8 g, 5.76 25 mmol), xantophos (360 mg, 0.623 mmol), Pd2dba3 (384 mg, 0.42 mmol), 4-bromochloro- 168 2-methylpyridazin-3(2H)-one (1.28 g, 5.76 mmol) and Cs2CO3 (5.05 g. 17.3 mmol) in 1,4- dioxane (40 mL) was heated at reflux for 2 h. After the completion of the reaction, the mixture was filtered off, and washed with MeOH (60 mL). The filtrate was evaporated in vacuo. The residue was purified on reverse phase Combi-flash to give 132a (1.3 g, 81%). 5 MS: [M+H]+ 282.
Example 132b (4-(5-(6,7-Dihydro-4H-pyrazolo[5,1-c][1,4]oxazinylamino)- 1-methyloxo-1,6-dihydropyridazinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)pyridinyl)methyl e 132b 10 Following the procedures as described for compound 131a, reaction of 3- xymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridin ylboronic acid 113i (200 mg, 0.52 mmol) and 132a (146 mg, 0.52 mmol) afforded 132b as a yellow solid (100 mg, 53%). LCMS: [M+H]+ 585 Example 132 5-(6,7-Dihydro-4H-pyrazolo[5,1-c][1,4]oxazin 15 ylamino)methyloxo-1,6-dihydropyridazinyl)(hydroxymethyl)pyridinyl)- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 132 Following the procedures as described for compound 131, hydrolysis of 132b (100 mg 0.171 mmol) with m hydroxide afforded 132 as a white solid (60 mg, 65%). LCMS: [M+H]+ 543. 1H NMR (500 MHz, CDCl3) δ 8.55 (d, J=5.0, 1H), 8.01 (s, 1H), 7.94 (s, 1H), 20 7.43 (d, J=5.5, 1H), 6.87 (s, 1H), 5.97 (s, 1H), 4.80 (s, 2H), 4.58 (s, 3H), 4.47 (s, 1H), 4.15- 1.14 (m, 2H), 4.11(s, 4H), 3.90 (s, 4H), 2.61-2.60 (m, 2H), 2.57 (t, J=6.5, 2H), 1.89-1.91 (m, 2H), 1.79-1.80 (m, 2H) Example 133a 6-Chloromethyl(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinylamino)pyridazin-3(2H)-one 133a 25 169 A 250-mL three-neck round-bottomed flask equipped with a reflux condenser, magnetic stirrer and nitrogen inlet was charged with 4-bromochloromethylpyridazin- 3(2H)-one (1.90 g, 8.53 mmol): , 5 113e (1.18 g, 7.75 mmol) and 1,4-dioxane (40 mL). The flask was purged with nitrogen and cooled to 0 ºC. A 1 M solution of lithium hexamethyldisilazide in THF (39 mL, 39.0 mmol) was added. After bubbling nitrogen through the resulting suspension for 30 min, Xantphos (381 mg, 0.659 mmol) and tris(dibenzylidene-acetone)dipalladium(0) (355 mg, 0.388 mmol) were added, and the on mixture was heated at reflux for 2 h. After this 10 time, the mixture was cooled to room temperature and diluted with water (10 mL). The pH of the solution was adjusted to 7.6 with 2 N hydrochloric acid. The organic layer was ted, and the aqueous layer was extracted with ethyl e (3 × 40 mL). The combined organic layers were dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography on silica to afford a 76% yield (1.74 g) of 133a as 15 an ite solid: mp 184–186 °C; 1H NMR (300 MHz, DMSO-d 6) δ 9.62 (s, 1H), 7.72 (s, 1H), 6.00 (s, 1H), 4.04 (t, 2H, J = 5.1 Hz), 3.65 (s, 3H), 3.53 (s, 2H), 2.82 (t, 2H, J = 5.1 Hz), 2.37 (s, 3H); MS (ESI+) m/z 295.1 (M+H).
Example 133b (4-(1-Methyl(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinylamino)oxo-1,6-dihydropyridazinyl)(1-oxo-3,4,6,7,8,9- 20 hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl Acetate 133b N N N NH AcO O N N N N O N 133b Following the procedures as described for compound 131a and ng with 3- (acetoxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridin ylboronic acid 113i (200 mg, 0.52 mmol) and 132a (153 mg, 0.52 mmol) afforded 132b as a 25 yellow solid (170 mg, 55%). LCMS: [M+H]+ 598 170 Example 133 2-(3-(Hydroxymethyl)(1-methyl(5-methyl-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)oxo-1,6-dihydropyridazinyl)pyridinyl)- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 133 Hydrolysis of 133b (160 mg 0.267 mmol) with lithium ide afforded 133 as a 5 white solid (94 mg, 63%). LCMS: [M+H]+ 556. 1H NMR (500 MHz, CDCl3) δ 8.55 (d, J=5.0, 1H), 7.98 (s, 1H), 7.89 (s, 1H), 7.43 (d, J=5.0, 1H), 6.87 (s, 1H), 5.94 (s, 1H), 4.57 (s, 3H), 4.47 (s, 1H), 4.11-4.15 (m, 4H), 3.89 (s, 3H), 3.87 (s, 1H), 3.61 (d, J=4.0 , 2H), 2.90 (s, 2H), , J=4.0, 2H), 2.57 (t, J=6.0, 2H), 2.49 (s, 3H), 1.89-1.91 (m, 2H), 1.79-.80 (m, 2H) 10 Example 134a 10-Bromo-1H,2H,3H,4H,6H,7H,8H,9H-pyrazino[1,2-a]indol- 1-one 134a Into a 250-mL 3-necked round-bottom flask was placed a solution of 3H,4H,6H,7H,8H,9H-pyrazino[1,2-a]indolone 101e(9.5 g, 49.94 mmol, 1.00 equiv) in N,N-dimethylformamide (100 mL), followed by the addition of N-bromosuccinimide (9.8 15 g, 55.06 mmol, 1.10 equiv) in several batches at 0oC. The resulting solution was stirred at room ature for 2 h and diluted with 500 mL of water. The precipitate was filtered and dried in a vacuum oven to afford 9.5 g (71%) of 119a as a light brown solid.
Example 134b 10-Fluoro-1H,2H,3H,4H,6H,7H,8H,9H-pyrazino[1,2-a]indol one 134b 20 Into a 2-L 4-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of 134a (40 g, 148.62 mmol, 1.00 equiv) in tetrahydrofuran (200 mL), followed by the addition of n-BuLi (2.4 M) (218 mL, 3.50 equiv) dropwise with stirring at -78 oC. The resulting solution was stirred at -40oC for 3 h. To this was added a solution of robenzenesulfonimide (98.7 g, 313.33 mmol, 2.10 equiv) in 25 tetrahydrofuran (200 mL) dropwise with stirring at -78oC. The ing solution was stirred at room temperature for 3 h, quenched by the addition of 200 mL of water and extracted with 3x500 mL of ethyl acetate. The combined organic layers were dried over anhydrous sodium e and concentrated under vacuum. The crude product (30 g) was purified by Prep-HPLC with the following conditions (mobile phase, A: 0.05% trifluoroacetic acid/water; B: CH3CN; 30 gradient: 10% B-25% B) to afford 5.05 g (16%) of 134b as a white solid. MS: [M+H]+ 209 . 1H NMR (300 MHz, CDCl3) δ 6.16 (br, 1H), .86 (m, 2H), 3.65-3.62 (m, 2H), 2.53- 2.47 (m, 4H), 1.88-1.80 (m, 2H), 1.77-1.72 (m, 2H).
Example 134c 4-Chloro(10-fluorooxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 134c 171 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and reflux ser was charged with 1,4-dioxane (60 mL), 134b (500 mg, 2.4 mmol): 5 2-bromochloronicotinaldehyde 103a(1.60 g, 7.2 mmol), and potassium acetate (471 mg, 4.8 mmol). After bubbling nitrogen through the resulting mixture for 30 minutes, os (140 mg, 0.24 mmol) and tris(dibenzylideneacetone)dipalladium(0) (220 mg, 0.24 mmol) were added, and the reaction mixture was heated at 80 ºC for 10 h. After this time the reaction was cooled to room temperature, partitioned between ethyl acetate (40 mL) and 10 water (40 mL), and filtered. The aqueous layer was separated and extracted with ethyl acetate (50 mL × 3). The combined organic layer was washed with brine (30 mL) and dried over sodium e. The drying agent was removed by filtration and the filtrate was concentrated under reduced re. The residue was purified on flash column eluting with 3:1 PE/EA to afford 134c (678 mg, 81%) as yellow solid. MS: [M+H]+ 348. 1H NMR (500 15 MHz, DMSO-d6) δ 9.87 (s, 1H), 8.60 (d, J=5.5, 1H), 7.56 (d, J=5.5, 1H), 4.23-4.25 (m, 2H), 4.13-4.15 (m, 2H), 2.59 (t, J=6.0, 2H), 2.41 (t, J=6.0, 2H), 1.75-1.80 (m, 2H), 1.66-1.70 (m, 2H) Example 134d 2-(10-Fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl)(1-methyl(5-(4-(oxetanyl)piperazinyl)pyridinylamino)oxo-1,6- 20 dihydropyridinyl)nicotinaldehyde 134d A mixture of 134c (300 mg, 0.86 mmol), yl(5-(4-(oxetanyl)piperazin yl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 101l 172 (403 mg, 0.86 mmol), Na (142 mg, 1.72 mmol), K3PO4 (460 mg, 1.72 mmol), PdCl2(dppf) (71 mg, 0.086 mmol) in CH3CN (25 mL) and H2O (1 mL) was heated at 100ºC for 3 hours. After reaction it was evaporated the residue was purified by silical-gel column eluting with methylene chloride/methanol (30:1) to afford 134d (312 mg, yield 55 %) as a 5 brown solid. MS: (M+H)+ 653.
Example 134 10-Fluoro(3-(hydroxymethyl)(1-methyl(5-(4-(oxetan- 3-yl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 134 To a solution of 134d (200 mg, 0.30 mmol) in MeOH (20 mL) was added NaBH4 (40 10 mg, 0.9 mmol). The mixture was stirred at 20 ºC for 2 h. After reaction it was evaporated and the residue was ed by reverse-phase prep-HPLC to afford 134 (108 mg, yield 54 %) as a yellow solid. MS: (M+H)+ 655. 1H NMR (500 MHz, DMSO-d6) δ 8.61 (d, J=2.0, 1H), 8.49 (d, J=5.0, 1H), 8.43 (s, 1H), 7.85 (d, J=2.5, 1H), 7.45 (d, J=1.5, 1H), 7.37-7.39 (m, 1H), 7.35 (d, J=5.0, 1H), 7.24 (d, J=9.0, 1H), 4.99 (s, 1H), 4.56 (t, J=6.5, 2H), 4.40-4.47 (m, 4H), 4.18- 15 4.22 (m, 2H), 4.05-4.09 (m, 1H), 3.84-3.96 (m, 1H), 3.60 (s, 3H), 3.41-3.46 (m, 1H), 3.07 (s, 4H), 2.54-2.61 (m, 2H), 2.39-2.42 (m, 6H), 1.78 (s, 2H), 1.69 (s, 2H) Example 135a yl(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 135a N N N NH O O N B O 135a 20 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a condenser was charged with compound 113h (1.0 g, 3 mmol), Pin2B2 (3.8 g, 15 mmol), Pd(dppf)Cl2 (137 mg, 0.15mmol), X-phos (143 mg, 0.3mmol), KOAc (88 mg, 9 mmol), and 1,4-dioxane (50 mL). After three cycles of vacuum/argon flush, the reaction mixture was heated at 60oC for 15 h. It was then cooled to room temperature and filtered. The filtrate was 25 concentrated under d pressure and the resulting residue was washed with petroleum ether to afford 135a as a yellow solid (0.87 g, 75%). MS: [M+H]+ 386 173 Example 135b 2-(10-Fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl)(1-methyl(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinylamino) 6-dihydropyridinyl)nicotinaldehyde 135b N N N NH O O N N N F O N 135b 5 A suspension of 135a (385 mg, 1 mmol), ro(10-fluorooxo-3,4,6,7,8,9- dropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 134c (347 mg, 1 mmol), K3PO4 (424 mg, 2 mmol), NaOAc (164g, 2mmol) and 1,1’- bis(diphenylphosphino)ferrocenedichloropalladium(II) (41 mg, 0.05 mmol) in CH3CN (50 ml) was heated at 100 °C under an N2 balloon for 4h. Analysis of on mixture by LCMS 10 showed completed conversion to the desired product. The reaction mixture was cooled to room temperature and diluted with DCM (50 ml) and water (80 mL). The aqueous layer was separated and extracted with DCM (3 × 50 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated. The dark residue was purified by silica gel column tography eluting with DCM/MeOH (from 80/1 to 30/1) to afford 135b (285 g, 50%) 15 as yellow solid. MS: [M+H]+ 571 Example 135 10-Fluoro(3-(hydroxymethyl)(1-methyl(5-methyl- 4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinylamino)oxo-1,6-dihydropyridinyl)pyridin- 2-yl)-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 135 To a solution of 135b (280 g, 0.49 mmol) in MeOH (50 mL) was added NaBH4 (56 g, 20 1.47 mmol) at room temerature. After the reaction was stirred for 3h, LCMS indicated the reaction was completed. Then the mixture was poured into H2O (50 mL) and extracted with DCM (50 mL × 3). The combined organic layer was washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by reverse-phase prep-HPLC to afford 135 (187 mg, 67%) as a white solid. MS: [M+H]+ 572. 1H NMR (500 MHz, CDCl3) δ 25 8.47 (d, J=5.5, 1H), 7.95 (d, J=2.0, 1H), 7.70 (d, J=2.0, 1H), 7.42 (s, 1H), 7.35 (d, J=5.5, 1H), 5.70 (s, 1H), 4.96 (t, J=7.0, 1H), 4.62 (s, 1H), 4.45 (s, 1H), 4.33 (s, 1H), 4.07-4.12 (m, 4H), 3.84 (s, 1H), 3.70 (s, 3H), 3.60 (s, 2H), 2.88 (t, J=5.5, 2H), 2.61 (s, 2H), 2.57 (s, 2H), 2.48 (s, 3H), 1.86-1.90 (m, 2H), 1.77 (s, 2H) 174 Example 136a (S)(10-Fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl) (1-methyl(5-(2-methyl(oxetanyl)piperazinyl)pyridin ylamino)oxo-1,6-dihydropyridinyl)nicotinaldehyde 136a 5 A 50 mL single-neck round-bottomed flask equipped with a magnetic r and a reflux condenser was charged with (S)methyl(5-(2-methyl(oxetanyl)piperazin yl) nylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 130f (225 mg, 1.5 eq., 0.47 mmol), 4-chloro(10-fluorooxo-3,4,6,7,8,9- hexahydropyrazino [1,2-a]indol-2(1H)-yl)nicotinaldehyde 134c (150 mg, 1 eq., 0.43 mmol): 10 PdCl2(dppf) (35 mg, 0.1 eq., 0.043 mmol), K3PO4 (273 mg, 3 eq., 1.29 mmol), and THF (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at reflux for 5 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by column tography with 15 DCM/EtOH (40:1) to afford 136a as yellow solid (100 mg, 34%). MS: [M+H]+ 667.3.
Example 136 (S)Fluoro(3-(hydroxymethyl)(1-methyl(5-(2- (oxetanyl) piperazinyl)pyridinylamino)oxo-1,6-dihydropyridin yl)pyridinyl)-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 136 A 25 mL single-neck round-bottomed flask equipped with a magnetic stirrer was 20 charged with 136a (100 mg, 1.0 eq., 0.15 mmol), NaBH4 (17 mg, 3.0 eq., 0.45 mmol), and MeOH (10 mL). The e was stirred at room temperature for 1 h. The residue was purified by reverse-phase prep-HPLC to afford 136 (64 mg, 64%). LCMS: [M+H]+ 669.3. 1H NMR (500 MHz, CDCl3) δ 8.64 (d, J=2.0, 1H), 8.48 (d, J=5.0, 1H), 7.96 (d, J=2.5, 1H), 7.83-7.82 (m, 2H), 7.36 (d, J=5.0, 1H), 7.30 (dd, J=2.5, 9.0, 1H), 6.81 (d, J=8.5, 1H), 4.99- 175 4.96 (m, 1H), 4.71-4.61 (m, 5H), 4.45-3.83 (m, 5H), 3.71 (s, 3H), 3.54-3.45 (m, 2H), 3.08- 3.06 (m, 2H), .47 (m, 7H), 2.21-2.17 (m, 1H), 1.89-1.76 (m, 4H), 0.98 (d, J=6.5, 3H) Example 137a (R)-(4-(1-Methyl(5-(2-methyl(oxetanyl)piperazin yl)pyridinylamino)oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- 5 hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl e 137a O N N N NH OAc O N N N O N 137a A mixture of bromomethyl(5-(2-methyl(oxetanyl)piperazin idinylamino)pyridin-2(1H)-one 151f, the enantiomer of 130f (283 mg, 0.65 mmol): 10 3-(acetoxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- yl)pyridinylboronic acid 113i (250 mg, 0.65 mmol), PdCl2(dppf) (53 mg, 0.065 mmol), NaOAc (107 mg, 1.3 mmol), K3PO4 (347 mg, 1.3 mmol) in acetonitrile (30 mL) was heated at 100oC for 3h. The solvent was evaporated in vacuo and the residue was purified by flash column chromatography eluting with 30:1 DCM/MeOH to afford 137a (216 mg, 48%) as a 15 brown solid. LCMS: [M+H]+ 693.4 Example 137 (R)(3-(hydroxymethyl)(1-methyl(5-(2-methyl (oxetanyl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 3,4,6,7,8,9-hexa-hydropyrazino[1,2-a]indol-1(2H)-one 137 To a solution of 137a (200 mg, 0.29 mmol) in propanol (8 mL), tetrahydrofuran (8 20 mL), and water (2.0 mL) was added LiOH (690 mg, 29 mmol). The mixture was stirred at 30 ℃ for 2 h. It was then evaporated and the residue was purified by reverse-phase prep- HPLC to afford 137 (143 mg, 76%) as a white solid. LCMS: (M+H)+ 651.4. 1H NMR (500 MHz, DMSO-d6) δ 8.63 (d, J=2.0, 1H), 8.49 (d, J=5.0, 1H), 8.45 (s, 1H), 7.84 (d, J=2.5, 1H), 7.47 (d, J=2.0, 1H), 7.37-7.39 (m, 1H), 7.35 (d, J=5.5, 1H), 7.25 (d, J = 9.5, 1H), 6.58 (s, 1H), 176 4.95 (t, J=4.0, 1H), 4.54-4.58 (m, 2H), 4.40-4.49 (m, 4H), 4.11-4.26 (m, 3H), 3.86-3.88 (m, 1H), 3.68 (s, 1H), 3.61 (s, 3H), .42 (m, 1H), 3.08-3.11 (m, 1H), 2.95 (t, J=9.0, 1H), 2.62-2.67 (m, 1H), 2.54-2.59 (m, 2H), 2.48 (t, J=6.0, 2H), .36 (m, 2H), 2.19 (t, J=8.0, 1H), 1.81 (s, 2H), 1.68-1.72 (m, 2H), 0.93 (d, J=6.0, 3H) 5 Example 138a 3-(1-Methyl(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinylamino)oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)isonicotinaldehyde 138a N N N NH CHO O N N N O N 138a A 100-mL single-neck bottomed flask was charged with 3-bromo(1-oxo- 10 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)isonicotinaldehyde 101f (298 mg, 0.7 mmol), 1-methyl(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinylamino) (4,4,5,5-tetra-methyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 135a (325 mg, 0.84 mmol), PdCl2(dppf) (30 mg, 0.035 mmol), K3PO4 (300 mg, 1.4 mmol), and NaOAc·3H2O (200 mg, 1.4 mmol) in CH3CN (70 mL). The system was evacuated and refilled with Argon. The 15 reaction e was heated at 100 ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by flash column chromatography eluting with 25:1 DCM/MeOH to afford 138a (220 mg, 55%) as a pale yellow solid. MS: [M+H]+ 553.3.
Example 138 2-(4-(Hydroxymethyl)(1-methyl(5-methyl-4,5,6,7- 20 tetrahydropyrazolo[1,5-a]pyrazinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 138 A mixture of 138a (200 mg, 0.36 mmol) and NaBH4 (50 mg, 1.2 mmol) in MeOH (60 mL) was stirred at room temperature for 2 h. The mixture was quenched with water and extracted with EtOAc (10 mL X 3). The combined EtOAc t was concentrated under 25 d pressure and the residue was purified with reverse-phase prep-HPLC to afford 138 (162 mg, 85%). LCMS: [M+H]+ :555.3. 1H NMR (500 MHz, CDCl3) δ 8.64 (s,1H), 8.49 (s, 1H), 7.97 (d, J=2.5, 1H), 7.42 (s, 1H), 7.33 (d, J=2, 1H), 6.88 (s, 1H), 5.68 (s, 1H), 4.65-4.63 ( m, 1H), 4.57-4.55 (m, 1H), 4.37 (t, J=11, 1H), 4.20-4.16 (m, 3H), 4.07-3.98 (m, 3H), 3.70 177 (s, 3H) , 3.59 (s, 2H), 2.87 (t, J=5.5, 2H), 2.61-2.56 (m, 4H), 2.48 (s, 3H), 1.92-1.90 (m, 2H), .79 (m, 2H) e 139a 4-Chloro(1-oxo-3,4,6,7,8,9-hexahydropyrido[3,4- b]indolizin-2(1H)-yl)nicotinaldehyde 139a 5 A 250-mL single-neck round-bottomed flask equipped with a magnetic r and reflux condenser was charged with 1,4-dioxane (50 mL), 2-bromochloronicotin-aldehyde 103a (1.4 g, 6.4 mmol), 3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin-1(2H)-one 112d (0.6 g, 3.2 mmol), Pd2(dba)3 (293 mg, 0.32mmol), XantPhos (370 mg, 0.64 mmol), and potassium 10 carbonate (627 mg, 6.4 mmol). After three cycles of vacuum/argon flush, the mixture was heated at 80 oC overnight. After this time the reaction was cooled to room temperature. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified by silica gel column chromatography eluting with DCM/CH3OH (20:1, V/V) to afford 139a (528 mg, 50%) as a yellow solid. MS: [M+H]+ 330. 1H NMR (500 MHz, CDCl3) δ 10.09 (s, 1H), 8.37 15 (d, J=5.5, 1H), 7.16 (d, J=5.5, 1H), 6.25 (s, 1H), 4.29-4.32 (m, 2H), 3.83-3.86 (m, 2H), 2.96- 2.99 (m, 2H), 2.75-2.78 (m, 2H), 2.00-2.07 (m, 2H), 1.82-1.85 (m, 2H) Example 139b 4-(1-Methyl(5-(4-(oxetanyl)piperazinyl)pyridin ylamino)oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrido[3,4- b]indolizin-2(1H)-yl)nicotinaldehyde 139b 20 A round-bottomed flask was charged with 139a (100 mg, 0.30 mmol), 1-methyl(5- etanyl)piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolan- 2-yl)pyridin-2(1H)-one 101l (140 mg, 0.30 mmol), PdCl2(dppf) (25 mg, 0.03 mmol), K3PO4.3H2O(160 mg, 0.60 mmol), NaOAc (59 mg, 0.60 mmol), acetonitrile (10 mL), and 25 H2O (5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100 oC for 3 h. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified on 178 flash column chromatography eluting with 1:3 petroleum/ethyl e to afford 139b as a yellow solid (95 mg, 50%). LCMS: [M+H]+ 635 Example 139 2-(3-(Hydroxymethyl)(1-methyl(5-(4-(oxetan yl)piperazinyl)pyridineylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 5 3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin-1(2H)-one 139 A mixture of 139b (95 mg, 0.15 mmol), NaBH4 (17 mg, 0.45), and CH3OH (10 mL) was stirred at 25 oC for 1 h. The mixture was extracted with CH2Cl2 (10 mL X 2). The combined CH2Cl2 extract was concentrated under reduced pressure. The residue was purified with reverse-phase prep-HPLC to afford 139 (60 mg, 63%). LCMS: [M+H]+ 637. 1H NMR 10 (500 MHz, DMSO-d6) δ 8.63 (d, J=2.0, 1H), 8.47 (d, J=5.5, 1H), 8.42 (s, 1H), 7.85 (d, J=2.5, 1H), 7.49 (d, J=2.0, 1H), 7.37-7.39 (m, 1H), 7.30 (d, J=5.0, 1H), 7.24 (d, J=9.0, 1H), 6.05 (s, 1H), 4.47-4.57 (m, 2H), 4.41-4.47 (m, 2H), 4.39-4.41 (m, 1H), 4.33-4.35 (m, 1H), 4.11-4.16 (m, 1H), .96 (m, 1H), 3.76-3.82 (m, 2H), 3.59 (s, 3H), 3.41-3.45 (m, 2H), .08 (m, 4H), .01 (m, 1H), 2.92-2.95 (m, 1H), 2.71-2.72 (m, 2H), .39 (m, 4H), 1.91-1.93 15 (m, 2H), 1.72-1.78 (m, 2H) Example 140a (S)(7,7-Difluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl) (1-methyl(5-(2-methyl(oxetanyl)piperazinyl)pyridin ylamino)oxo-1,6-dihydropyridinyl)nicotinaldehyde 140a 20 Following the procedures as described in Example 130g, reaction of (S)methyl (5-(2-methyl(oxetanyl)piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2- dioxaborolanyl)pyridin-2(1H)-one 130f and 4-chloro(7,7-difluorooxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)nicotin-aldehyde (170 mg): 179 afforded 140a was as a yellow solid (200 mg, 60%). LCMS: [M+H]+ 684.3. 4-Chloro(7,7- difluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)nicotin-aldehyde was prepared from 7,7-difluoro-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one according to the reaction scheme in Figure 25. 5 Example 140 (S)-7,7-Difluoro(3-(hydroxymethyl)(1-methyl(5-(2- methyl(oxetan- 3-yl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridin yl)pyridinyl)-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 140 Following the procedures as described in Example 130, sodium borohydride reduction of 140a (200 mg) afforded 140 as a yellow solid (104 mg, 51%). LCMS: [M+H]+ 686.3. 1H 10 NMR (500 MHz, DMSO) δ 8.62 (d, J=2.0, 1H), 8.46-8.49 (m, 2H), 7.83 (d, J=3.0, 1H), 7.45(d, J=2.5, 1H), 7.35-7.38(m, 2H), 7.25 (d, J=9.5, 1H), 6.64 (s, 1H), 4.95-4.97 (m, 1H), .57 (m, 2H), 4.38-4.48 (m, 4H), 4.15-4.27 (m, 3H), .90 (m, 1H), 3.67 (s, 1H), 3.59 (s, 3H), 3.26-3.39 (m, 3H), 3.08-3.10 (m, 1H), 2.92-2.96 (m, 1H), 2.63-2.67 (m, 2H), 2.52-2.55 (m, 1H), 2.30-2.36 (m, 2H),2.18-2.24 (m, 3H), 0.93 (d, J=6.0, 3H) 15 Example 141a 4-Chloro{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 141a A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with 2-bromochloronicotinaldehyde 103a (3.0 g, 13.6 20 mmol), 4,4-dimethyl-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienone 107e (1.84 g, 9.0 mmol), tris(dibenzylideneacetone)dipalladium(0) (826 mg, 0.9 mmol), XantPhos (1.04 mg, 1.8 mmol), Cs2CO3 (5.8 g, 18.0 mmol), and oxane (40 mL). After three cycles of vacuum/argon flush, the mixture was heated at 90 oC for 5 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the 25 resulting e was recrystallized from ethyl acetate to afford 141a as yellow solid (730 mg, 31.7 %). MS: [M+H]+ 344.0.
Example 141b -Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dien yl}[1-methyl({5-methyl-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin yl}amino)oxo-1,6- dihydropyridinyl]pyridinecarbaldehyde 141b 180 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with 141a (130mg, 0.38 mmol), 1-methyl(5-methyl-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolan 5 yl)pyridin-2(1H)-one 135a (146 mg, 0.38 mmol), f)Cl2 (31 mg, 0.038 mmol), K3CO3 (105 mg, 0.76 mmol), and DMF (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at 110 oC for 2 h. It was then cooled to room temperature and ed.
The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (30:1) to afford 10 141b as brown solid (160 mg, 74.6 %). MS: [M+H]+ 567.3.
Example 141 2-[3'-Hydroxymethylmethyl(5-methyl-4,5,6,7-tetrahydropyrazolo [1,5-a]pyrazinylamino)oxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl]-7,7-dimethyl- 3,4,7,8-tetrahydro-2H,6H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazinone 141 To a solution of 141b (150 mg, 0.26 mmol) at room temperature in methanol (10 mL) 15 was added sodium borohydride (29 mg, 0.78 mmol) and the resulting mixture was stirred for 30 minutes. It was ed with water (1.0 mL) and concentrated. The residue was purified by reverse-phase PLC to afford 141 (35 mg, 23.2 %). LCMS: [M+H]+ 569.3. 1H NMR (500 MHz, CDCl3) δ 8.46 (d, J = 5.0, 1H), 7.94 (d, J = 2.5, 1H), 7.72 (d, J = 2.0, 1H), 7.41 (s, 1H), 7.33 (d, J = 5.5, 1H), 6.83 (s, 1H), 5.68 (s, 1H), 5.03-5.00 (m, 1H), 4.64-4.61 (m, 1H), 20 4.51-4.48 (m, 1H), 4.32-4.27 (m, 1H), 4.21-4.09 (m, 4H), 3.91-3.82 (m, 1H), 3.69 (s, 3H), 3.62-3.58 (m, 2H), 2.87 (t, J=2.5, 2H), 2.57 (d, J=4.0, 2H), 2.54 (s, 2H), 2.51 (s, 3H), 1.27 (s, 6H) Example 142a 5-Bromomethyl(pyrimidinylamino)pyridin-2(1H)-one 142a 25 A 100-mL single-neck round-bottomed flask ed with a magnetic stirrer and nitrogen inlet was charged with 3,5-dibromomethylpyridin-2(1H)-one (2.00 g, 21.0 mmol), 181 2-aminopyrimidine (5.61 g, 21.0 mmol), cesium carbonate (13.7 g, 42.1 mmol), DMF (5 mL) and 1,4-dioxane (70 mL). After bubbling en through the resulting suspension for 30 min, Xantphos (1.10 g, 1.89 mmol) and tris(dibenzylideneacetone)dipalladium(0) (963 mg, 1.05 mmol) were added. A reflux condenser was attached to the flask, and the reaction 5 mixture was heated at 100 °C for 4 h. After this time, the mixture was cooled to room temperature and diluted with 90:10 methylene chloride/methanol (150 mL) and water (100 mL), and the layers were separated. The aqueous layer was extracted with 90:10 methylene chloride/methanol (50 mL), and the combined organic layers were washed with brine and dried over sodium sulfate. The drying agent was removed by filtration. The filtrate was 10 concentrated under reduced pressure, and the resulting residue was purified by flash column chromatography a, 90:10 methylene chloride/methanol) to afford 142a in 58% yield (3.42 g) as an amorphous light green solid: mp 217–219 °C; 1H NMR (500 MHz, CDCl3) δ 9.29 (s, 1H), 8.77 (s, 1H), 8.72 (d, J = 2.5 Hz, 1H), 8.36 (d, J = 6.0 Hz, 1H), 7.69 (d, J = 2.5 Hz, 1H), 7.37 (dd, J = 5.5, 1.0 Hz, 1H), 3.53 (s, 3H); LCMS (ESI+) m/z 281.0 (M+H). 15 e 142b (4-(1-Methyloxo(pyrimidinylamino)-1,6- dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridin- ethyl e 142b A sealed tube equipped with a magnetic stirrer was charged with 142a (154.5 mg, 20 0.55 mmol), oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)(4,4,5,5- tetramethyl-1,3,2-dioxaborolanyl)pyridinyl)methyl acetate 113i (252.5 mg, 0.55 mmol), Pd(dppf)Cl2 (25.9 mg, 0.03135 mmol), NaOAc (108 mg, 1.1 mmol), K3PO4·3H2O (293 mg, 1.1 mmol), acetonitrile (6 mL), and water ( 0.5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 110 oC for 2 h. It was then filtered and the te was evaporated 25 in vacuo. The residue was purified by silica gel column chromatography eluting with dichloromethane/methanol (15:1, V/V) to afford 142b (117 mg, 30%) as a brown solid.
LCMS: [M+H]+ 540.2 Example 142 2-[3'-Hydroxymethylmethyloxo(pyrimidin ylamino)-1,6-dihydro-[3,4']bipyridinyl-2'-yl]-3,4,6,7,8,9-hexahydro-2H-pyrazino[1,2-a]indol- 30 1-one 142 182 A e of 142b (121.6 mg, 0.225 mmol) and LiOH (100 mg, 4.2 mmol) in iPrOH/THF (1:1, 4 mL) and H oC for 0.5 h. The mixture was 2O (1 mL) was stirred at 35 evaporated in vacuo and the residue was extracted with EtOAc (20 mL X 3). The combined EtOAc extract was concentrated under reduced re and the residue was purified by 5 reverse-phase prep-HPLC to afford 142 (54 mg, 48.2%) as a pale yellow solid. LCMS: [M+H]+ 498.1. 1H NMR (500 MHz, DMSO-d6) δ 9.23 (s, 1H), 8.76 (d, J=2.5, 1H), 8.65 (s, 1H), 8.50 (d, J=5.0, 1H), 8.31 (d, J=6.0, 1H), 7.69 (d, J=2.5, 1H), 7.37 (d, J=5.0, 1H), 7.31- 7.33 (m, 1H), 6.58 (s, 1H), 4.97 (t, J=4.5, 1H), 4.39-4.43 (m, 2H), 4.10-4.24 (m, 3H), 3.87 (d, J=12.0, 1H), 3.61 (s, 3H), 2.57-2.64 (m, 2H), 2.47 (d, J=6, 2H), 1.79 (d, J=4.0, 2H), 1.69 (d, 10 J=6.0, 2H) Example 143a 1-Methyl(pyrimidinylamino)(4,4,5,5-tetramethyl- 1,3,2-dioxaborolanyl)pyridin-2(1H)-one 143a A 250-mL single-neck round-bottomed flask equipped with a ic stirrer and a 15 condenser was charged with 5-bromomethyl(pyrimidinylamino)pyridin-2(1H)-one 142a (4.0 g, 14 mmol), X-phos (400 mg, 0.7 mmol), Pd2(dba)3 (635 mg, 0.7 mmol), KOAc (7.3 mg, 28 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (10.6 g, 42 mmol), and 1,4-dioxane (100 mL). After three cycles of vacuum/argon flush, the reaction e was heated at 60 oC for 8 h. It was then cooled to room temperature and filtered. The 20 filtrate was concentrated under reduced pressure and the resulting residue was ed by flash column chromatography eluting with 5:1 petroleum ether/ethyl acetate to afford 143a as a pale yellow solid (3.8 mg, 82%). MS: [M+H]+ 329.5.
Example 143b 4-(1-Methyl(pyrimidinylamino)oxo-1,6- dihydropyridinyl){4,4-dimethyloxothiaazatricyclo[6.4.0.02,6]dodeca- 25 1(8),2(6)-dienyl}nicotinaldehyde 143b 183 A 250-mL single-neck round-bottomed flask equipped with a ic stirrer and a condenser was d with 143a (150 mg, 0.46 mmol), 4-chloro{4,4-dimethyloxo thiaazatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienyl}pyridinecarbaldehyde 109a 5 (164 mg, 0.46 mmol): , Pd(dppf)Cl2 (16 mg, 0.02mmol), K3PO4.3H2O (223 mg, 0.92 mmol) in CH3CN (5 mL) and H2O (1 mL). After three cycles of vacuum/argon flush, the reaction mixture was heated at 100oC for 3 h. It was then cooled to room temperature and filtered. The filtrate was 10 concentrated under reduced pressure and the resulting residue was purified by flash column chromatography g with 20:1 of DCM/MeOH to afford 143b as a yellow solid (110 mg, 48%). MS: [M+H]+ 527.
Example 143 6-[3'-Hydroxymethylmethyloxo(pyrimidin ylamino)-1,6-dihydro-[3,4']bipyridinyl-2'-yl]-2,2-dimethyl-2,3,5,6-tetrahydro-1H,4Hthia- 15 6-aza-cyclopenta[a]indenone 143 A mixture of 143b (110 mg, 0.2 mmol), NaBH4 (30 mg, 0.8 mmol), and MeOH (5 mL) was stirred at 25 oC for 30 mins. The mixture was evaporated in vacuo and the residue was extracted with EtOAc (10 mL X 2). The combined EtOAc extract was concentrated under reduced pressure and the residue was purified with reverse-phase prep-HPLC to afford 20 143 (48 mg, 44%). LCMS: [M+H]+ 529. 1H NMR (500 MHz, DMSO) δ 9.23 (s, 1H), 8.76 (d, J=2.5, 1H), 8.65 (s, 1H), 8.51-8.49 (m, 1H), 8.31 (m, 1H), 7.67 (d, J=3.0, 1H), 7.38-7.37 (m, 1H), .31 (m, 1H), 5.02-5.01 (m, 1H), 4.43 (d, J=2.5, 2H), 4.18-4.15 (m, 1H), 3.83- 3.81 (m, 1H), 3.61-3.59 (m, 3H), 3.03-2.99 (m, 1H), 2.91-2.89 (m, 1H), 2.76 (s, 2H), 2.60- 2.53 (m, 2H), 1.23-1.22 (m, 6H) 184 Example 144a ethyl(pyrimidinylamino)oxo-1,6- dihydropyridinyl){4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dien- nicotinaldehyde 144a 5 A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a condenser was charged with 1-methyl(pyrimidinylamino)(4,4,5,5-tetramethyl-1,3,2- dioxaborolanyl)pyridin-2(1H)-one 143a (150 mg, 0.46 mmol), 4-chloro{4,4-dimethyl- 9-oxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 108a (157 mg, 0.46 mmol): 10 , f)Cl2 (16 mg, 0.02 mmol), K3PO4.3H2O (223 mg, 0.92 mmol) in CH3CN (5 mL) and H2O (1 mL). After three cycles of vacuum/argon flush, the reaction mixture was heated at 100oC for 3 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by flash column 15 chromatography eluting with 20:1 of DCM/MeOH to afford 144a as a yellow solid (98 mg, 48%). MS: [M+H]+ 510.
Example 144 2-[3'-Hydroxymethylmethyloxo(pyrimidinylamino)-1,6- dihydro-[3,4']bipyridinyl-2'-yl]-7,7-dimethyl-3,4,7,8-tetrahydro-2H,6H-cyclopenta- [4,5]pyrrolo[1,2-a]pyrazinone 144 20 A mixture of 144a (98 mg, 0.19 mmol), NaBH4 (30 mg, 0.8mmol) and MeOH (5 mL) was stirred at 25oC for 30 mins. The e was evaporated in vacuo and the residue was extracted with EtOAc (10 mL x 2). The combined EtOAc extract was concentrated under reduced pressure and the residue was purified with reverse-phase prep-HPLC to give 144 (25 mg, 42%). LCMS: [M+H]+ 512. 1H NMR (500 MHz, DMSO) δ 9.18 (s, 1H), 8.76-8.74 (m, 25 1H), 8.64 (s, 1H), 8.50-8.47 (m, 1H), 8.31-8.30 (m, 1H), 7.68-7.69 (m, 1H), 7.37-7.36 (m, 185 1H), 7.33-7.31 (m, 1H), 6.56 (s, 1H), 5.07-5.04 (m, 1H), 4.44-4.41 (m, 2H), 4.23-4.18 (m, 3 H), 3.86-3.84 (m, 1H), 3.61 (s, 3H), .56 (m, 2H), 2.42 (s, 2H), .20 (m, 6H) Example 145a (S)- 4-(1-methyl(5-(2-methyl(oxetanyl)piperazin yl)pyridinylamino)oxo-1,6-dihydropyridinyl){4,4-dimethyloxothia 5 azatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienyl}nicotinaldehyde 145a A 50 mL single-neck bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with (S)methyl(5-(2-methyl(oxetanyl) piperazin yl) pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 10 130f (160 mg, 1 eq., 0.33 mmol), 4-chloro{4,4-dimethyloxothia cyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienyl}pyridinecarbaldehyde 109a (120 mg, 1 eq., 0.33 mmol): , PdCl2(dppf) (27 mg, 0.1 eq., 0.033 mmol), K3PO4 (140 mg, 2 eq., 0.66 mmol), 15 NaOAc (54 mg, 2 eq., 0.66 mmol), and CH3CN (20 mL). After three cycles of vacuum/argon flash, the mixture was heated at 100oC for 2 h. It was then cooled to room temperature and ed. The filtrate was concentrated under reduced pressure and the resulting residue was purified by column chromatography eluting with DCM/EtOH (40/1) to afford 145a as yellow solid (97 mg, 43%). MS: [M+H]+ 680.3. 20 Example 145 6-{3'-Hydroxymethylmethyl[5-((S)methyloxetan yl-piperazinyl)-pyridinylamino]oxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}-2,2- dimethyl-2,3,5,6-tetrahydro-1H,4Hthiaaza-cyclopenta[a]indenone 145 A 25 mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with 145a (97 mg, 1.0 eq., 0.14 mmol), NaBH4 (16 mg, 3.0 eq., 0.42 mmol), and 25 MeOH (10 mL). The mixture was stirred at room temperature for 1 h. The residue was 186 purified by reverse-phase prep-HPLC to afford 145 (61 mg, 63%). LCMS: [M+H]+ 682.3. 1H NMR (500 MHz, CDCl3) δ 8.65 (d, J=2.5, 1H), 8.50 (d, J=5.0, 1H), 7.97 (d, J=2.5, 1H), 7.84 (s, 1H), 7.80 (d, J=2.5, 1H), 7.37 (d, J=5.0, 1H), 7.30 (dd, J=3.0, 9.0, 1H), 6.81 (d, J=9.0, 1H), 4.82-4.79 (m, 1H), 4.71-4.61 (m, 5H), 4.45-4.31 (m, 2H), 3.85-3.80 (m, 1H), 5 3.71 (s, 3H), 3.54-3.46 (m, 2H), 3.07 (d, J=5.0, 2H), 2.98-2.93 (m, 2H), 2.79 (s, 2H), 2.60- 2.46 (m, 5H), 2.21-2.18 (m, 1H), 1.28 (s, 6H), 0.98 (d, J=6.0, 3H) e 146a 4-Chloro(10-fluorooxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 146a 10 To a solution of ochloronicotinaldehyde 103a (1600 mg, 7.27 mmol), 10- fluoro-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one (500 mg, 2.40 mmol): in dioxane (50 mL) was added KOAc (471 mg, 4.82 mmol), Pd2(dba)3 (220 mg, 0.24 mmol), and 4,5-bis(diphenylphosphino)-9,9-dimethyl-xanthene (140 mg, 0.24 mmol). After 15 bubbling argon h the resulting solution for 30 min, the mixture was d at 80oC for 10 h. It was allowed to cool to room temperature and H2O (100 mL) was added. The aqueous layer was separated and extracted with ethyl acetate (2 × 200 mL). The combined organic layer was washed with brine (100 mL) and dried over sodium sulfate. The drying agent was removed by filtration and the filtrate was concentrated under reduced pressure. The residue 20 was purified on flash column eluting with PE/EA (3:1) to afford 146a as a yellow solid (420 mg, 49%). LCMS: [M+H]+ 348 Example 146b 2-(10-Fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl)(1- methyloxo(pyrimidinylamino)-1,6-dihydropyridin yl)nicotinaldehyde 146b 187 A round-bottomed flask was charged with 146a (200 mg, 0.58 mmol), 1-methyl (pyrimidinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl) n-2(1H)-one 143a (227 mg, 0.69 mmol), PdCl2(dppf) (47 mg, 0.06 mmol), K3PO4 (244 mg, 1.15 mmol), 5 NaOAc (94 mg, 1.15 mmol), acetonitrile (30 mL), and H2O (3 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 3 h. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified on flash column chromatography eluting with 1:20 methanol/dichloro-methane to afford 146b as a red solid (79 mg, 27%).
LCMS: [M+H]+ 514 10 Example 146 10-Fluoro(3-(hydroxymethyl)(1-methyloxo (pyrimidinylamino) -1,6-dihydropyridinyl)pyridinyl)-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-1(2H)-one 146 A mixture of 146b (79 mg, 0.15 mmol), NaBH4 (22 mg, 0.60), and CH3OH (10 mL) was stirred at 25 oC for 1 h. The mixture was extracted with CH2Cl2 (10 mL X 2). The 15 combined CH2Cl2 t was concentrated under reduced pressure. The residue was purified with reverse-phase prep-HPLC to afford 146 (39 mg, 49%). LCMS: [M+H]+ 516. 1H NMR (500 MHz, CDCl3) δ 8.83 (d, J=2.0, 1 H), 8.78 (s, 1 H), 8.52 (d, J=5.0, 1 H), 8.35 (d, J=5.5, 1 H), 8.12 (s, 1 H), 8.03 (d, J=2.0, 1 H), 7.36 (d, J=5.0, 1 H), 6.76-6.77 (m, 1 H), 5.07 (s, 1 H), 4.65 (d, J=9.5, 1 H), 4.48 (d, J=9.5, 1 H), 4.29 (d, J=1.5, 1 H), .13 (m, 2 H), 3.79 (d, 20 J=6.5, 1 H), 3.73 (s, 3 H), 2.52-2.58 (m, 4 H), 1.85-1.90 (m, 2 H), 1.77 (d, J=5.0, 2 H) Example 147a 4-Chloro(1-oxo-3,4,6,7,8,9-hexahydropyrido[3,4- b]indolizin-2(1H)-yl)nicotinaldehyde 147a A 250-mL -neck round-bottomed flask equipped with a magnetic stirrer and 25 reflux condenser was charged with 1,4-dioxane (50 mL), 2-bromochloronicotin-aldehyde 103a (1.4 g, 6.4 mmol), 3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin-1(2H)-one 112d (0.6 g, 188 3.2 mmol), Pd2(dba)3 (293 mg, 0.32 mmol), XantPhos (370 mg, 0.64 mmol), and potassium acetate (627 mg, 6.4 mmol). After three cycles of /argon flush, the mixture was heated at 80 oC overnight. After this time the reaction was cooled to room temperature. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified by silica gel 5 column chromatography eluting with CH2Cl2/CH3OH (20:1, V/V) to afford 147a (528 mg, 50%) as a yellow solid. MS: [M+H]+ 330. 1H NMR (500 MHz, CDCl3) δ 10.09 (s, 1H), 8.37 (d, J=5.5, 1H), 7.16 (d, J=5.5, 1H), 6.25 (s, 1H), 4.29-4.32 (m, 2H), 3.83-3.86 (m, 2H), 2.96- 2.99 (m, 2H), 2.77-2.78 (m, 2H), 2.00-2.07 (m, 2H), 1.83-1.85 (m, 2H) Example 147b 2-(3-Formyl(1-methyl(5-methyl-4,5,6,7- 10 tetrahydropyrazolo[1,5-a]pyrazinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin-1(2H)-one 147b A round-bottomed flask was charged with 4-chloro(1-oxo-3,4,6,7,8,9- hexahydropyrido[3,4-b]indolizin-2(1H)-yl)nicotinaldehyde 147a (100 mg, 0.30 mmol), 1- 15 methyl(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinylamino)(4,4,5,5- ethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 135a (116 mg, 0.30 mmol), PdCl2(dppf) (25 mg, 0.03 mmol), K3PO4.3H2O(160 mg, 0.60 mmol), NaOAc (59 mg, 0.60 mmol), itrile (10 mL), and H2O (5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 3 h. It was then filtered and the filtrate was evaporated in 20 vacuo. The residue was purified on flash column tography eluting with 1:3 petroleum/ethyl acetate to afford 147b as a yellow solid (100 mg, 60%). LCMS: [M+H]+ 553 Example 147 2-(3-(Hydroxymethyl)(1-methyl(5-methyl-4,5,6,7- ydro-pyrazolo[1,5-a]pyrazinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin-1(2H)-one 147 25 A mixture of 147b (100 mg, 0.18 mmol), NaBH4 (21 mg, 0.54), and CH3OH (10 mL) was stirred at 25oC for 1 h. The mixture was extracted with CH2Cl2 (10 mL X 2). The ed CH2Cl2 extract was concentrated under reduced pressure. The residue was purified with reverse-phase prep-HPLC to afford 147 (60 mg, 60%). LCMS: [M+H]+ 555. 1H NMR 189 (500 MHz, DMSO) δ 8.45 (d, J=5.0, 1H), 8.19 (s, 1H), 8.06 (d, J=5.0, 1H), 7.41 (d, J=2.0, 1H), 7.29 (d, J=5.0, 1H), 6.04 (s, 1H), 5.88 (s, 1H), 4.92 (s, 1H), 4.33-4.42 (m, 2H), 4.11-4.16 (m, 1H), 3.91-3.96 (m, 3H), 3.77-3.82 (m, 2H), 3.57 (s, 3H), 3.45-3.48 (m, 2H), 2.91-3.01 (m, 2H), 2.71-2.79 (m, 4H), 2.35 (s, 3H), 1.90-1.92 (m, 2H), 1.71-1.79 (m, 2H) 5 Example 148a 3-(2-Bromochloropyridinyl)oxetanol 148a To a on of ochloropyridine (14 g, 70 mmol) in dry THF (200 mL) was added LDA (42.0 mL, 84.0 mmol, 2.0 M) dropwise at -70 ºC. After stirring for 0.5 h at this temperature, a solution of oxetanone (6.6 g, 90 mmol) in dry THF (40 mL) was added 10 slowly and the reaction mixture was stirred at 0 0C for an additional 1 h. Saturated aqueous NH4CI (50 mL) and EA (200 mL) were added. The mixture was partitioned between water and ethyl acetate. The aqueous layer was extracted with ethyl acetate. The combined organic layer was washed with water and brine. After drying over ous magnesium sulfate, the solvent was evaporated and the crude material was purified by SGC eluting with DCM) to 15 afford 148a as a yellow solid (8.8 g, 45%). MS: [M+H]+ 266.0.
Example 148b 2-(4-Chloro(3-hydroxyoxetanyl)pyridinyl)-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-1(2H)-one 148b A 100mL single-neck round-bottomed flask equipped with a magnetic stirrer was 20 charged with 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 101e (190 mg, 1.0 mmol), 148a (795 mg, 3.0 mmol), CuI (95 mg, 0.5 mmol), DMEDA (88 mg, 1.0 mmol), KOAc(294 mg, 3.0 mmol), and 1,4-dioxane (50 ml). The system was evacuated and then ed with N2.
A reflux condenser was attached to the flask, and the on mixture was heated at 85 ºC for 15 h. Then, the mixture was cooled to room temperature and filtered. The filtrate was 25 concentrated under reduced re and the resulting residue was purified by flash column chromatography eluting with 2:1 petroleum ether/ethyl acetate to afford 148b as a yellow solid (156 mg, 42%). MS: [M+H]+ 374.2. 190 Example 148 (S)(3-(3-Hydroxyoxetanyl)(1-methyl(5-(2-methyl- 4-(oxetanyl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridin yl)-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 148 A 100-mL single-neck round-bottomed flask was charged with 148b (100 mg, 0.3 5 mmol), (S)methyl(5-(2-methyl(oxetanyl)piperazinyl)pyridineylamino) (4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 130f (173 mg, 0.36 mmol), Pd(dppf)Cl2 (15 mg, 0.015 mmol), K3PO4 (130 mg, 0.6 mmol), and NaOAc·3H2O (90 mg, 0.6 mmol) in CH3CN (30 mL). The system was evacuated and refilled with N2. The reaction mixture was heated at 100 ºC for 2 h. It was then cooled to room temperature and ed. 10 The filtrate was concentrated under reduced pressure and the resulting residue was ed by flash column chromatography eluting with 25:1 DCM/MeOH to afford 148 (30 mg, 20 %) as a pale yellow solid. MS: [M+H]+ 693.3. 1H NMR (500 MHz, CDCl3) δ 8.69 (d, J=2, 1H), 8.50 (d, J=5, 1H), 8.01 (d, J=2.5, 1H), 7.85 (s, 1H), 7.67 (s, 1H), 7.38-7.32 (m, 2H), 6.89 (s, 1H), 6.83 (d, J=8.5, 1H), 6.67 (s, 1H), 4.93 (d, J=6, 1H), .63 (m, 6H), 4.46 (d, J=7.5, 15 1H), 4.24-4.18 (m, 2H), 4.10-4.05 (m, 1H), 3.90 (d, J=12.5, 1H), 3.70 (s, 3H), 3.55-3.46 (m, 2H), 3.10 (t, J=4.5, 2H), 2.63-2.48 (m, 7H), 2.22 (t, J=7.5, 1H), 1.92-1.88 (m, 2H), 1.82-1.77 (m, 2H), 1.02-1.00 (m, 3H) Example 149a (S)(1-Methyl(5-(2-methyl(oxetanyl)piperazin yl)pyridinylamino)oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- 20 hexahydropyrido[3,4-b]indolizin-2(1H)-yl)nicotinaldehyde 149a A 100-mL round-bottomed flask equipped with a reflux ser was charged with 4-chloro(1-oxo-3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin-2(1H)-yl)nicotinaldehyde 139a (100 mg, 0.30 mmol), (S)methyl(5-(2-methyl(oxetanyl)-piperazin 25 yl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 191j (146 mg, 0.30 mmol), PdCl2(dppf) (25 mg, 0.030 mmol), trihydrate (160 mg, 0.60 mmol), sodium acetate (49 mg, 0.60 mmol), acetonitrile (20 mL), and water (3 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 3 h. It was then 191 filtered and the filtrate was evaporated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 1:3 petroleum/ethyl acetate to afford 149a as a yellow solid (97 mg, 50%). MS-ESI: [M+H]+ 649 Example 149 2-{3'-Hydroxymethylmethyl[5-((S)methyloxetanyl- 5 piperazinyl)-pyridinylamino]oxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}-2,3,5,6,7,8- hexahydro-4H-2,4b-diaza-fluorenone 149 A mixture of 149a (97 mg, 0.15 mmol), NaBH4 (17 mg, 0.45), and methanol (10 mL) was stirred at 25oC for 1 h. The reaction mixture was then quenched with water (10 mL) and concentrated under reduced pressure. The e was extracted with dichloromethane (2 X 10 10 mL). The combined dichloromethane extract was concentrated under reduced pressure and the residue was purified with reverse-phase prep-HPLC to afford 149 (62 mg, 63%). MS-ESI: [M+H]+ 651.3. 1H NMR (500 MHz, DMSO-d6) δ 8.63 (s, 1H), 8.46 (d, J = 5.0 Hz, 1H), 8.43 (s, 1H), 7.83 (d, J = 3.0 Hz, 1H), 7.48 (d, J = 2.0 Hz, 1H), 7.38-7.36 (m, 1H), 7.31 (J = 5.0 Hz, 1H), 7.25-7.23 (m, 1H), 6.04 (s, 1H), 4.57-4.55 (m, 2H), 4.48-4.46 (m, 1H), 4.42-4.38 (m, 15 2H), 4.35-4.33 (m, 1H), .12 (m, 1H), 3.96-3.94 (m, 1H), 3.82-3.78 (m, 2H), 3.69-3.67 (m, 1H), 3.59 (s, 3H), 3.41-3.38 (m, 2H), 3.18-3.15 (m, 2H), .95 (m, 3H), 2.73-2.71 (m, 2H), 2.30-2.28 (m, 2H), 2.20-2.16 (m, 1H), 1.93-1.89 (m, 3H), 1.77-1.75 (m, 1H), 0.93 (d, J = 6.5 Hz, 3H).
Example 150a -Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- 20 dienyl}[1-methyl({5-methyl-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazinyl}amino) 6-dihydropyridinyl]pyridinecarbaldehyde 150a N N N NH O O N N N O N 150a A 25-mL single-neck round-bottomed flask equipped with a magnetic r and a reflux condenser was charged with 3-bromo{4,4-dimethyloxo-1,10- 25 ricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 107f (233 mg, 0.60 mmol), 1-methyl({5-methyl-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazinyl}amino) (4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)-1,2-dihydropyridinone 135a (231 mg, 0.60 mmol), Pd(dppf)Cl2 (49 mg, 0.060 mmol), potassium acetate (118 mg, 1.2 mmol), 192 K3PO4.trihydrate (320 mg, 1.2 mmol), itrile (12 mL), and water ( 5 . After three cycles of vacuum/argon flush, the mixture was heated at 110oC for 2 h. It was then filtered and the filtrate was evaporated under reduced pressure. The e was purified by silica-gel column tography g with 30:1 dichloromethane/methanol to afford 150a (168 mg, 5 49%) as a solid. MS-ESI: [M+H]+ 567 Example 150 2-[4-Hydroxymethyl-1'-methyl-5'-(5-methyl-4,5,6,7-tetrahydropyrazolo [1,5-a]pyrazinylamino)-6'-oxo-1',6'-dihydro-[3,3']bipyridinylyl]-7,7-dimethyl- 3,4,7,8-tetrahydro-2H,6H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazinone 150 To a solution of 150a (170 mg, 0.30 mmol) in methanol (10 mL) was added sodium 10 borohydride (68 mg, 1.8 mmol) at 0oC. The mixture was stirred at room temperature for 30 minutes. Then the reaction mixture was quenched with water (2 mL) and concentrated. The residue was purified with reverse-phase prep-HPLC to afford 150 (42 mg, 25%) as a pale yellow solid. MS-ESI: [M+H]+ 569. 1H NMR (500 MHz, CDCl3) δ 8.63 (s, 1H), 8.48 (s, 1H), 7.95 (d, J = 2.0 Hz, 1H), 7.40 (s, 1H), 7.32 (d, J = 2.5 Hz, 1H), 6.82 (s, 1H), 5.67 (s, 1H), 15 4.63-4.55 (m, 2H), 4.37-4.35 (m 1H), 4.22-4.18 (m, 3H), 4.05-3.97 (m, 3H), 3.69 (s, 3H), 3.59-3.57 (m, 2H), 2.86 (t, J = 6.0 Hz, 2H), 2.56 (s, 2H), 2.50 (s, 2H), 2.46 (s, 3H), 1.26 (s, 6H).
Example 151a (R)-tert-Butyl 3-Methyl(6-nitropyridinyl)piperazine carboxylate 151a 20 A 250-mL single-neck round-bottomed flask equipped with a magnetic r and a reflux condenser was charged with 1,4-dioxane (60 mL), 5-bromonitropyridine (2.0 g, 10.0 mmol), (R)-tert-butyl 3-methylpiperazinecarboxylate (2.0 g, 10.0 mmol), and cesium carbonate (6.5 g, 20 mmol). After bubbling nitrogen through the resulting mixture for 10 25 minutes, tris(dibenzylideneacetone)dipalladium(0) (915 mg, 1.0 mmol) and XantPhos (579 mg, 1.0 mmol) were added. The system was ted to three cycles of vacuum/argon flush and heated at 100 oC for 15 h. After this time the reaction was cooled to room temperature and filtered. The filtrate was partitioned between ethyl acetate (100 mL) and water (100 mL). 193 The aqueous layer was ted and extracted with ethyl acetate (3 x 50 mL). The combined organic layer was washed with brine (100 mL) and dried over sodium sulfate. The drying agent was removed by tion and the filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 30:1 5 romethane/methanol to afford 151a (1.6 g, 44%) as yellow solid. MS-ESI: [M+H]+ 323. 1H NMR (500 MHz, DMSO-d δ 8.21 (d, J = 3.5 Hz, 1H), 8.18 (d, J = 9.0 Hz, 1H), 7.45- 6) 7.43 (m, 1H), .33 (m, 1H), 3.92-3.99 (m, 1H), 3.80 (d, J = 12.5 Hz, 2H), 3.06-3.23 (m, 3H), 1.43 (s, 9H), 1.09 (d, J = 6.5 Hz, 3H).
Example 151b (R)-tert-Butyl 4-(6-Aminopyridinyl)methylpiperazine 10 carboxylate 151b A 250-mL flask was purged with nitrogen and charged with 151a (1.5 g, 4.6 mmol), 10% ium on carbon (50% wet, 200 mg), and methanol (70 mL). It was evacuated, charged with hydrogen gas, and stirred at room temperature for 10 h. The hydrogen was then ted and nitrogen was charged into the flask. The catalyst was removed by filtration 15 through a pad of CELITE® and the filtrate was concentrated under reduced pressure to afford 151b (1.1 g, 81%) as brown solid. MS-ESI: [M+H]+ 293 Example 151c (R)-tert-Butyl 4-(6-(5-Bromomethyloxo-1,2- opyridinylamino)pyridinyl)methylpiperazinecarboxylate 151c A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 20 reflux condenser was charged with 1,4-dioxane (40 mL), 151b (1.0 g, 3.4 mmol), 3,5- dibromomethylpyridin-2(1H)-one (2.7 g, 10.2 mmol), and cesium carbonate (2.2 g, 6.8 mmol). After bubbling nitrogen through the resulting mixture for 10 minutes, XantPhos (198 mg, 0.34 mmol) and tris(dibenzylideneacetone)dipalladium(0) (313 mg, 0.34 mmol) were added. The reaction mixture was subjected to three cycles of vacuum/argon flush and heated 25 at 100oC for 5 h. After this time the reaction was cooled to room temperature and filtered.
The filtrate was partitioned between ethyl acetate (50 mL) and water (50 mL). The aqueous layer was separated and extracted with ethyl acetate (3 X 30 mL). The ed organic layer was washed with brine (50 mL) and dried over sodium sulfate. The drying agent was removed by filtration and the filtrate was concentrated under reduced pressure. The residue 30 was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 151c as yellow solid (1.1 g, 63%). MS-ESI: [M+H]+ 478. e 151d (R)Bromomethyl(5-(2-methylpiperazinyl)pyridin ylamino)pyridin-2(1H)-one 151d 194 To a mixture of 151c (600 mg, 1.26 mmol) in methanol (20 mL) was added HCl/dioxane (4M, 4 mL). The reaction mixture was stirred at room temperature for 4 h. It was then concentrated under reduced re. The residue was ed with aqueous 1M NaOH and extracted with dichloromethane (3 X 30 mL). The ed organic layer was 5 washed with brine and concentrated under reduced pressure to afford 151d (450 mg, 95%) as yellow solid. MS-ESI: [M+H]+ 378.
Example 151e (R)Bromomethyl(5-(2-methyl(oxetan yl)piperazinyl)pyridinylamino)pyridin-2(1H)-one 151f A mixture of 151d (40.0 g, 106 mmol), oxetanone (11.4 g, 159 mmol), NaBH3CN 10 (10.0 g, 159 mmol), and zinc chloride (21.3 g, 159 mmol) in methanol (700 mL) was stirred at 50oC for 5 hours. water (50 mL) was added to the mixture and concentrated under reduced pressure. The residue was extracted with dichloromethane (3 X 200 mL) and the combined organic layer was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 40:1 dichloromethane/methanol to afford 151e (35 g, 15 73%). MS: [M+H]+ 434.
Example 151f (R)Methyl(5-(2-methyl(oxetanyl)piperazinyl)pyridin ylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 151f To a solution of 151e (2.0 g, 4.60 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2- 20 dioxaborolane) (3.50 g, 13.80 mmol) in dioxane (50 mL) was added PdCl2(dppf) (377.10 mg, 0.46 mmol) and potassium acetate (2.70 g, 27.80 mmol). The mixture was stirred at 10 oC for 12 h under argon atmosphere. After reaction the mixture was filtered and the te was evaporated under reduced pressure. The residue was ed by -gel column chromatography eluting with 15:1 methylene chloride/methanol to afford 151f (1.10 g, 49%) 25 as a brown solid. MS: [M+H]+ 482.3 195 Example 151g (R)(1-Methyl(5-(2-methyl(oxetanyl)piperazin yl)pyridinylamino)oxo-1,6-dihydropyridinyl)(1-oxo-6,7,8,9- tetrahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 151g A 100-mL round-bottomed flask equipped with a reflux condenser was charged with 5 4-chloro(1-oxo-6,7,8,9-tetrahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 103b (150 mg, 0.45 mmol), 151f (331 mg, 0.69 mmol), dppf) (37 mg, 0.045 mmol), K3PO4 (190 mg, 0.90 mmol), sodium acetate (74 mg, 0.90 mmol), acetonitrile (15 mL), and water (1.5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100 oC for 3 h.
It was then filtered and the te was ated under d pressure. The residue was 10 purified with silica-gel column chromatography eluting with 1:20 methanol/dichloromethane to afford 151g as a red solid (89 mg, 30%). MS-ESI: [M+H]+ 647 Example 151 2-{3'-Hydroxymethylmethyl[5-((R)methyloxetanylpiperazinyl )-pyridinylamino]oxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}-6,7,8,9- tetrahydro-2H-pyrazino[1,2-a]indolone 151 15 A mixture of 151g (89 mg, 0.14 mmol), NaBH4 (22 mg, 0.60), and methanol (10 mL) was stirred at 25oC for 1 h. The e was quenched with water (8 mL) and concentrated under reduced pressure. The residue was extracted with dichloromethane (2 X 10 mL). The combined dichloromethane extract was concentrated under reduced pressure and the residue was purified with reverse-phase prep-HPLC to afford 151 (35 mg, 39%). MS-ESI: [M+H]+ 20 649. 1H NMR (500 MHz, CDCl3) δ 8.65 (d, J = 2.0 Hz, 1H), 8.55 (d, J = 5.0 Hz, 1H), 8.46 (s, 1H), 7.83 (d, J = 2.5 Hz, 1H), 7.50-7.48 (m, 2H), 7.38-7.36 (m, 1H), 7.26-7.24 (m, 2H), 6.83-6.80 (m, 2H), 4.98 (bs, 1H), 4.57-4.54 (m, 2H), 4.48-4.33 (m, 4H), 3.67-3.66 (m, 1H), 3.60 (s, 3H), 3.39-3.38 (m, 2H), 3.09-3.08 (m, 1H), 2.96-2.94 (m, 1H), 2.76-2.74 (m, 2H), .62 (m, 2H), 2.36-2.31 (m, 2H), 2.20-2.17 (t, J = 7.5 Hz, 1H), 1.88-1.86 (m, 2H), 1.75- 25 1.74 (m, 2H), 0.93 (d, J = 6.5 Hz, 3H).
Example 152a tert-Butyl 8-(6-Nitropyridinyl)-3,8- diazabicyclo[3.2.1]octanecarboxylate 152a 196 Boc Boc Boc N Br N N Pd/C N NH N EtOH, rt, 16 h N NO2 Pd2(dba)3, Xantphos, N NH2 N NO2 Cs2CO3, dioxane, 110°C, O/N 152a 152b Boc HN Br N N O N N HCl/dioxane N NH Br N NH rt, 5 h; O O N Pd2(dba)3, Xantphos, N Br Cs2CO3, dioxane, Br 120 °C, sealed, 2 h 152d 152c O O N N N N O N NH N NH 113h O AcO O O N ZnCl2, NaCNBH3, N N N Pd(dppf)Cl2, MeOH, 50 °C, 5 h Br K3PO4, NaOAc O N MeCN, H2O, 152e reflux, 3 h 152f A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 1,4-dioxane (100 mL), 5-bromonitropyridine (2.5 g, 12.4 mmol), tert-butyl 3,8-diazabicyclo[3.2.1]octanecarboxylate(869 g, 4.1 mmol), 5 Pd2(dba)3 (193 mg, 0.21 mmol), os (237 mg, 0.41 mmol), and cesium carbonate (2.7 g, 8.2 mmol). After three cycles of vacuum/argon flush, the e was stirred at 110oC overnight. The on was cooled to room ature. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified by silica-gel column chromatography eluting with 3:1 petroleum ether/ethyl acetate to afford 152a (2.63 g, 66.8%) as a yellow solid. 10 MS-ESI: [M+H]+ 335.2.
Example 152b tert-Butyl 8-(6-Aminopyridinyl)-3,8- diazabicyclo[3.2.1]octanecarboxylate 152b 197 A 100-mL single-neck round-bottomed flask was purged with nitrogen and charged with 152a (2.5 g, 7.5 mmol), 10% ium on carbon (50% wet, 250 mg) and methanol (40 mL). The mixture was evacuated, charged with hydrogen gas, and stirred at room ature for 16 h. The hydrogen was then evacuated and nitrogen was charged into the flask. The 5 catalyst was removed by filtration through a pad of ® and the filtrate was concentrated under reduced pressure to afford 152b (1.51 g, 66%) as a ess oil. MS-ESI: [M+H]+ 305.3 Example 152c tert-Butyl 5-Bromomethyloxo-1,2-dihydropyridin- 3-ylamino)pyridinyl)-3,8-diazabicyclo[3.2.1]octanecarboxylate 152c 10 A sealed tube equipped with a magnetic stirrer was charged with 152b (1.3 g, 4.3 mmol), 3,5-dibromomethylpyridin-2(1H)-one (1.2 g, 4.3 mmol), tris(dibenzylideneacetone)dipalladium(0) (394 mg, 0.43 mmol), XantPhos (497 mg, 0.86 mmol), Cs2CO3 (2.8 g, 8.6 mmol), and oxane (15 mL). After three cycles of vacuum/argon flush, the mixture was stirred at 120oC for 2 h. The filtrate was concentrated 15 under reduced pressure and the resulting residue was purified by -gel column chromatography eluting with 50:1 dichloromethane/methanol to afford 152c as a yellow solid (900 mg, 43%). MS-ESI: [M+H]+ 490.3.
Example 152d 3-(5-(3,8-Diazabicyclo[3.2.1]octanyl)pyridinylamino) bromomethyl-pyridin-2(1H)-one 152d 20 A mixture of 152c (900 mg, 1.84 mmol) and 4.0M HCl/dioxane (60 mL) was stirred at room temperature for 5 h. It was then concentrated under reduced pressure to afford crude 152d as a yellow solid (700 mg, 98%), which was used in the next step without further purification. MS-ESI: [M+H]+ 390.3.
Example 152e 5-Bromomethyl(5-(3-(oxetanyl)-3,8- 25 diazabicyclo[3.2.1]octanyl)pyridinylamino)pyridine-2(1H)-one 152e A mixture of 152d (676 mg, 1.7 mmol), oxetanone (251 mg, 3.5 mmol), NaBH3CN (274 mg, 4.4 mmol), and zinc chloride (592 mg, 4.4 mmol) in methanol (30 mL) was stirred at 50oC for 5 hours. water was added and the e was concentrated under reduced pressure. The residue was extracted with dichloromethane three times. The combined extract 30 was concentrated under reduced pressure to afford crude 152e as a yellow solid (650 mg, 84%), which was used in the next step without further purification. MS-ESI: [M+H]+ 446.2.
Example 152f Methyl(5-(3-(oxetanyl)-3,8- diazabicyclo[3.2.1]octanyl)pyridineylamino)oxo-1,6-dihydropyridinyl)(1-oxo- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl Acetate 152f 198 A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 152e (300 mg, 0.67 mmol), 3-(acetoxymethyl)(1-oxo- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinylboronic acid 113h (257 mg, 0.67 mmol), Pd(dppf)Cl2 (55 mg, 0.067 mmol), K3PO4 (284 mg, 1.34 mmol), sodium acetate 5 (110 mg, 1.34 mmol), water (6 , and acetonitrile (20 mL). After three cycles of vacuum/argon flush, the mixture was stirred at reflux for 3 h. It was then cooled to room temperature and filtered. The filtrate was trated under reduced pressure and the resulting residue was purified by -gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 152f as a brown solid (200 mg, 42%). : [M+H]+ 10 705.4.
Example 152 2-{3'-Hydroxymethylmethyl[5-((1S,5R)oxetanyl-3,8-diazabicyclo [3.2.1]octyl)-pyridinylamino]oxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}- 3,4,6,7,8,9-hexahydro-2H-pyrazino[1,2-a]indolone 152 A mixture of 152f (180 mg, 0.26 mmol) and lithium hydroxide (215 mg, 5.1 mmol) in 15 i-propanol/THF (1:1, 4 mL) and water (1 mL) was stirred at 35oC for 1 h. The e was evaporated in vacuo and the residue was diluted with water and ethyl acetate. The water phase was separated and extracted with ethyl acetate (2 X 10 mL). The combined ethyl acetate extract was concentrated under reduced re and the residue was purified by reverse-phase prep-HPLC to afford 152 (12 mg, 71%) as a yellow solid. MS-ESI: [M+H]+ 20 663.3. 1H NMR (500 MHz, DMSO-d6) δ 8.55 (d, J = 2.5 Hz, 1H), 8.48 (d, J = 5.0 Hz, 1H), 8.32 (s, 1H), 7.80 (d, J = 2.5 Hz, 1H), 7.43 (d, J = 2.0 Hz, 1H), 7.33 (d, J = 5.0 Hz, 1H), 7.25-7.23 (m, 1H), 7.20 (d, J = 9.0 Hz, 1H), 6.57 (s, 1H), 4.96-4.94 (m, 1H), .43 (m, 3H), 4.39-4.37 (m, 3H), 4.25-4.19 (m, 5H), 3.85 (d, J = 11.5 Hz, 1H), 3.59 (s, 3H), 2.66-2.54 (m, 4H), 2.40-2.36 (m, 3H), 2.17 (d, J = 10.5 Hz, 2H), 1.94-1.65 (m, 8H). 25 Example 153a 2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}[1-methyl({5-[(2R)methyl(oxetanyl)piperazinyl]pyridin- 2-yl}amino)oxo-1,6-dihydropyridinyl]pyridinecarbaldehyde 153a O N N N NH O O N N N O N 153a 199 A 50-mL round-bottomed flask equipped with a reflux condenser was d with 4- chloro{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 108a (105 mg, 0.30 mmol), 1-methyl({5-[(2R)methyl(oxetan yl)piperazinyl]pyridinyl}amino)(tetra-methyl-1,3,2-dioxaborolanyl)-1,2- 5 dihydropyridinone 151g (216 mg, 0.45 mmol), PdCl2(dppf) (25 mg, 0.030 mmol), K3PO4 (126 mg, 0.60 mmol), sodium acetate (49 mg, 0.60 mmol), acetonitrile (15 mL), and water (1.5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 80oC for 2 h. It was then filtered and the filtrate was evaporated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 1:20 methanol/dichloromethane to 10 afford 153a as a red solid (82 mg, 41%). MS-ESI: [M+H]+ 663 Example 153 2-{3'-Hydroxymethylmethyl[5-((R)methyloxetanylpiperazinyl )-pyridinylamino]oxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}-7,7-dimethyl- 3,4,7,8-tetrahydro-2H,6H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazinone 153 A mixture of 153a (82 mg, 0.12 mmol), NaBH4 (22 mg, 0.60), and methanol (10 mL) 15 was stirred at 25 oC for 1 h. It was then quenched with water (5 mL) and concentrated under reduced pressure. The residue was extracted with dichloromethane (2 X 10 mL). The combined dichloromethane extract was concentrated under d pressure and the residue was purified with reverse-phase prep-HPLC to afford 153 (22 mg, 28%). MS-ESI: [M+H]+ 665. 1H NMR (500 MHz, CDCl3) δ 8.65 (d, J = 2.0 Hz, 1H), 8.48 (d, J = 5.0 Hz, 1H), 7.96 20 (s, 1H), 7.84 (s, 1H), 7.83 (s, 1H),7.36 (d, J = 5.0 Hz, 1H), .26 (m,1H), 6.84-6.80 (m, 2H), 5.30 (s, 1H), 4.71-4.32 (m, 7H), 4.15 (d, J = 5.0 Hz, 2H), 3.85 (t, J = 8.0 Hz, 1H), 3.71 (s, 3H), .43 (m, 2H), .06 (m, 2H), 2.57-2.48 (m, 7H), .20 (m, 1H), 1.27 (s, 6H), 0.98 (d, J = 6.5 Hz, 3H).
Example 154a 4-[1-Methyl({5-[(2R)methyl(oxetanyl)piperazin 25 yl]pyridinyl} amino)oxo-1,6-dihydropyridinyl]{6-oxothia-4,5- diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridinecarbaldehyde 154a 200 A 100-mL round-bottomed flask equipped with a reflux condenser was charged with 4-chloro{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridine- 3-carbaldehyde 124a (84 mg, 0.24 mmol), methyl(5-(2-methyl(oxetan yl)piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin- 5 2(1H)-one 151g (173 mg, 0.36 mmol), dppf) (20 mg, 0.024 mmol), K3PO4 (100 mg, 0.48 mmol), sodium acetate (40 mg, 0.48 mmol), acetonitrile (20 mL), and water (2 mL).
After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 3 h. It was then filtered and the filtrate was evaporated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 1:20 methanol/dichloromethane to afford 10 154a as a red solid (112 mg, 70%). MS-ESI: [M+H]+ 665 Example 154 Hydroxymethylmethyl[5-((R)methyloxetanylpiperazinyl )-pyridinylamino]oxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}-6,7,8,9- tetrahydro-3H-benzo[4,5]thieno[2,3-d]pyridazinone 154 A mixture of 154a (150 mg, 0.23 mmol), NaBH4 (35 mg, 0.92), and methanol (10 15 mL) was stirred at room temperature for 1 h. The mixture was then quenched with water (8 mL) and concentrated under reduced pressure. The residue was extracted with dichloromethane (2 x 10 mL). The combined romethane extract was concentrated under reduced pressure and the residue was purified with reverse-phase prep-HPLC to afford 154 (29 mg, 19%). MS-ESI: [M+H]+ 667. 1H NMR (500 MHz, DMSO-d6) δ 8.64 (d, J = 2.5 20 Hz, 1H), 8.56 (d, J = 5.0 Hz, 1H), 8.49-8.47 (m, 2H), 7.85 (d, J = 3.0 Hz, 1H), 7.53 (d, J = 5.0 Hz, 1H), 7.48 (d, J = 2.5 Hz, 1H), 7.38-7.36 (m, 1H) , 7.24 (d, J = 8.5 Hz, 1H), 4.85 (t, J = 9.5 Hz, 1H), 4.57-4.54 (m, 2H), .36 (m, 4H), .68 (m, 1H), 3.60 (s, 3H), 3.40- 3.36 (m, 1H), 3.11-3.07 (m, 1H), 2.97-2.86 (m, 6H), 2.33-2.31 (m, 2H), 2.16 (t, J = 8.5 Hz, 1H), 1.89-1.86 (m, 4H), 0.92 (d, J = 6.5 Hz, 3H). 25 Example 155a 3-(5-((2S,5R)-2,5-Dimethyl(oxetanyl)piperazin yl)pyridinylamino)methyl(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin- 2(1H)-one 155a A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was d with 5-bromo(5-((2S,5R)-2,5-dimethyl(oxetan 30 erazinyl)pyridinylamino)methylpyridin-2(1H)-one 122e (3.0 g, 6.70 mmol), Pin2B2 (8442 mg, 33.5 mmol), Pd2(dba)3 (311 mg, 0.34 mmol), X-phos (319 mg, 0.67 mmol), potassium acetate (1970 mg, 20.1mmol), and dioxane (50 mL). After three cycles of vacuum/argon flush, the mixture was heated at 60 ºC for 16 h. It was then cooled to room temperature and ed. The filtrate was concentrated under reduced pressure. The resulting 201 residue was washed with 8:1 petroleum ethyl acetate (80 mL) to afford 155a as a yellow solid (3 g, 90%). MS: [M+H]+ 496.4. e 155b 4-(5-(5-(2,5-Dimethyl(oxetanyl)piperazinyl)pyridin ylamino)methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- 5 hexahydropyrido[3,4-b]indolizin-2(1H)-yl)nicotinaldehyde 155b O O N N N N N NH N NH 139a O N O O O N Pd(dppf)Cl2, K3PO4, H2O N N B NaOAC,CH3CN 100 °C, 2h O N O 155a 155b A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 4-chloro(1-oxo-3,4,6,7,8,9-hexahydropyrido[3,4- b]indolizin-2(1H)-yl)nicotinaldehyde 139a (133 mg, 0.40 mmol), 155a (198 mg, 0.40 mmol), 10 Pd(dppf)Cl2 (17 mg, 0.020 mmol), K3PO4 (254 mg, 1.2 mmol), sodium acetate (98 mg, 1.2 mmol), water (5 drops), and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the e was heated at reflux for 3 h. It was then cooled to room temperature and filtered.
The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 15 155b as white solid (80 mg, 30%). MS-ESI: [M+H]+ 663.3.
Example 155 2-{5-[5-((2S,5R)-2,5-Dimethyloxetanyl-piperazinyl)-pyridin- 2-ylamino]-3'-hydroxymethylmethyloxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}- 2,3,5,6,7,8-hexahydro-4H-2,4b-diaza-fluorenone 155 To a solution of 155b (80 mg, 0.12 mmol) at 0oC in ol (5 mL) was added 20 sodium borohydride (12 mg, 0.36 mmol). The on mixture was stirred for 30 minutes. It was then quenched with water (1 mL) and concentrated under reduced pressure. The residue was purified by reverse-phase prep-HPLC to afford 155 (32 mg, 40 %). MS-ESI: [M+H]+ 665.3. 1H NMR (500 MHz, CDCl3) δ 8.68 (d, J = 2.0 Hz, 1H), 8.50 (d, J = 5.0 Hz, 1H), 8.04 (d, J = 2.5 Hz, 1H), 7.88 (s, 1H), 7.86 (d, J = 2.5 Hz, 1H), 7.37-7.33 (m, 2H), 6.82 (d, J = 9.0 25 Hz, 1H), 6.32 (s, 1H), 5.02 (d, J = 13.0 Hz, 1H), 4.78-4.71 (m, 2H), 4.67-4.61 (m, 3H), 4.44 - 4.39 (m,1H), 4.31-4.29 (m,1H), 3.96-3.91 (m, 1H), .80 (m, 2H), 3.78-3.75 (m, 1H), 202 3.72 (s, 3H), 3.21-3.19 (m, 1H), 3.01-2.93 (m, 3H), 2.85-2.83 (m, 2H), 2.72 (d, J = 10.0 Hz, 2H), 2.49-2.47 (m, 1H), 2.05-2.03 (m, 2H), 1.98-1.97 Example 156a 4-(5-(5-((2S,5R)-2,5-Dimethyl(oxetanyl)piperazin yl)pyridinylamino)methyloxo-1,6-dihydropyridinyl)(10-fluorooxo- 5 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 156a O N N N NH O O N N N F O N 156a A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with 3-(5-((2S,5R)-2,5-dimethyl(oxetanyl)piperazin yl)pyridinylamino)methyl(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin- 10 2(1H)-one 155a (171 mg, 0.35 mmol), 4-chloro(10-fluorooxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 134c (120 mg, 0.35 mmol), K3PO4 (146 mg, 0.69 mmol), PdCl2(dppf) (28 mg, 0.035 mmol), sodium acetate (56 mg, 0.69 mmol), water (5 drops) and acetonitrile (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at 90ºC for 2 h. It was then cooled to room temperature and filtered. The 15 filtrate was concentrated under reduced pressure and the resulting e was purified by silica-gel column chromatography eluting with 40/1 dichloromethane/methanol to afford 156a as a yellow solid (60 mg, 25%). : [M+H]+ 681.3 Example 156 2-{5-[5-((2S,5R)-2,5-Dimethyloxetanyl-piperazinyl)-pyridin- 2-ylamino]-3'-hydroxymethylmethyloxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}fluoro- 20 3,4,6,7,8,9-hexahydro-2H-pyrazino[1,2-a]indolone 156 A 50-mL round-bottomed flask ed with a magnetic stirrer was charged with 156a(60 mg, 0.088 mmol), NaBH4 (17 mg, 0.44 mmol), and methanol (10 mL). The e was stirred at room temperature for 1 h. It was then quenched with water and concentrated under reduced pressure. The residue was ed by reverse-phase prep-HPLC to afford 156 25 (15 mg, 25%). MS-ESI: [M+H]+ 683.5. 1H NMR (500 MHz, CDCl3) δ 8.67 (d, J = 2.5 Hz, 1H), 8.48 (d, J = 5.0 Hz, 1H), 8.02 (d, J = 2.5 Hz, 1H), 7.87 (s, 1H), 7.84 (d, J = 2.0 Hz, 1H), 7.37-7.35 (m, 2H), 6.81 (d, J = 9.0 Hz, 1H), 4.99-4.59 (m, 6H), 4.45-4.32 (m, 2H), 4.12-4.03 203 (m, 2H), 3.85-3.73 (m, 2H), 3.71 (s, 3H), 3.19-3.16 (m, 1H), 2.91-2.89 (m, 1H), 2.75-2.69 (m, 2H), 2.57-2.47 (m, 5H), 1.97-1.76 (m, 5H), 0.89-0.87 (m, overlap, 6H).
Example 157 2-{5'-[5-((2S,5R)-2,5-Dimethyloxetanyl-piperazinyl)-pyridin- ino]hydroxymethyl-1'-methyl-6'-oxo-1',6'-dihydro-[3,3']bipyridinylyl}- 5 3,4,6,7,8,9-hexahydro-2H-pyrazino[1,2-a]indolone 157 A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 3-bromo(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2a]indol-2(1H)-yl)isonicotinaldehyde 101f (200 mg, 0.54 mmol), 3-(5- ((2S,5R)-2,5-dimethyl(oxetanyl)piperazinyl)pyridinylamino)methyl(4,4,5,5- 10 tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 155a (267 mg, 0.54 mmol), Pd(dppf)Cl2 (44 mg, 0.054 mmol), K3PO4 (229 mg, 1.08 mmol), sodium acetate (89 mg, 1.08 mmol), water (0.2 mL) and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at reflux for 2 h. It was then cooled to room temperature and filtered.
The filtrate was concentrated under d pressure and the residue was purified by reverse- 15 phase prep-HPLC to afford 157 (35.5 mg, 11%) as a yellow solid. MS-ESI: [M+H]+ 665.4. 1H NMR (500 MHz, CDCl 3) δ .63 (m, 2H), 8.50 (s, 1H), 7.80 (s, 1H), 7.87 (s, 1H), 7.48-7.47 (m, 1H), 7.36 (d, J = 7.5 Hz, 1H), 6.88 (s, 1H), 6.81 (d, J = 9.0 Hz, 1H), 4.75-4.54 (m, overlap, 6H), 4.37-4.13 (m, overlap, 4H), .95 (m, 1H), 3.74-3.73 (m, 1H), 3.72 (s, 3H), 3.19-3.15 (m, 1H), 2.91-2.90 (m, 1H), 2.74-2.44 (m, overlap, 7H), 1.92-1.88 (m, 2H), 20 .79 (m, 2H), 0.90-0.89 (m, 6H).
Example 158a 4-[5-({5-[(2S,5R)-2,5-Dimethyl(oxetanyl)piperazin idinyl}amino)methyloxo-1,6-dihydropyridinyl]{4,4-dimethyloxo- 1,10-diazatri-cyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 158a O N N N NH O O N N N O N 158a 25 A 100-mL single-neck round-bottomed flask equipped with a reflux condenser was charged with (4-chloro{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- dienyl}pyridinecarbaldehyde) 108a (280 mg, 0.80 mmol), 3-(5-((2S,5R)-2,5-dimethyl- 4-(oxetanyl)piperazinyl)pyridinylamino)methyl(4,4,5,5-tetramethyl-1,3,2- 204 dioxaborolanyl)pyridin-2(1H)-one 155a (480 mg, 0.96 mmol), Pd(dppf)Cl2 (33 mg, 0.040 mmol), K3PO4 (339 mg, 1.6 mmol), sodium acetate·trihydrate (218 mg, 1.6 mmol), and acetonitrile (100 mL). The system was evacuated and refilled with N2. The reaction mixture was heated at 100ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate 5 was concentrated under reduced pressure and the resulting residue was ed by silica-gel column chromatography eluting with 25:1 of dichloromethane/methanol to afford 158a (300 mg, 54%) as a yellow brown solid. MS-ESI: [M+H]+ 677.3.
Example 158 2-{5-[5-((2S,5R)-2,5-Dimethyloxetanyl-piperazinyl)-pyridin- 2-ylamino]-3'-hydroxymethylmethyloxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}-7,7- 10 dimethyl-3,4,7,8-tetrahydro-2H,6H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazinone 158 A mixture of 158a (200 mg, 0.30 mmol) and NaBH4 (36 mg, 0.90 mmol) in methanol (30 mL) was stirred at 30oC for 1 h. The mixture was quenched with water (5 mL) and extracted with ethyl acetate (3 X 10 mL). The combined ethyl acetate t was concentrated under reduced pressure and the residue was purified by reverse-phase prep- 15 HPLC to afford 158 (110 mg, 55%) as a white solid. MS-ESI: [M+H]+ 679.4. 1H NMR (500 MHz, CDCl3) δ 8.68 (d, J = 2.0 Hz, 1H), 8.50 (d, J = 5.0 Hz, 1H), 8.03 (d, J = 2.5 Hz, 1H), 7.88 (s, 1H), 7.87 (d, J = 2.0 Hz, 1H), 7.37-7.36 (m, 2H), 6.85 (s, 1H), 6.81 (d, J = 3.5 Hz, 1H), 5.07 (t, J = 7.0 Hz, 1H), 4.77-4.71 (m, 2H), 4.67-4.61 (m, 3H), 4.53-4.51 (m, 1H), 4.34- 4.32 (m, 1H), 4.16 (d, J = 5.5 Hz, 2H), 3.88-3.86 (m, 1H), 3.76 (t, J = 7.5 Hz, 1H), 3.72 (s, 20 3H), .17 (m, 1H), 2.92 (dd, J = 3.0, 11.5 Hz, 1H), 2.76-2.70 (m, 2H), 2.58 (d, J = 6.0 Hz, 2H), 2.52 (s, 2H), 2.49-2.46 (m, 1H), 1.97-1.93 (m, 1H), 1.28 (s, 6H), 0.92-0.89 (m, 6H). e 159a 4-[5-({5-[2,5-Dimethyl(oxetanyl)piperazinyl]pyridin- 2-yl}amino)methyloxo-1,6-dihydropyridinyl]{6-oxothia-4,5- diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridinecarbaldehyde 159a O N N N NH O O N S N N O N 25 159a A 100-mL round-bottomed flask equipped with a reflux condenser was charged with ro{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridine- 3-carbaldehyde 124a (200 mg, 0.58 mmol), 3-({5-[(2S,5R)-2,5-dimethyl(oxetan 205 yl)piperazinyl]pyridinyl}amino)methyl(4,4,5,5-tetramethyl-1,3,2-dioxaborolan yl)-1,2-dihydropyridinone 155a (1.0 g, 2.0 mmol), PdCl2(dppf) (47 mg, 0.060 mmol), K3PO4 (280 mg, 1.2 mmol), sodium acetate (95 mg, 1.2 mmol), acetonitrile (15 mL), and water (1.5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC 5 for 3 h. It was then filtered and the filtrate was evaporated under reduced re. The residue was purified with -gel column chromatography eluting with 1:20 methanol/dichloromethane to afford 159a as a red solid (150 mg, 38%). MS-ESI: [M+H]+ 679 Example 159 3-{5-[5-((2S,5R)-2,5-Dimethyloxetanyl-piperazinyl)-pyridin- 10 2-ylamino]-3'-hydroxymethylmethyloxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}-6,7,8,9- tetrahydro-3H-benzo[4,5]thieno[2,3-d]pyridazinone 159 A e of 159a (130 mg, 0.19 mmol), NaBH4 (22 mg, 0.60), and methanol (10 mL) was stirred at 25 oC for 1 h. The mixture was then quenched with water (8 mL) and concentrated under reduced pressure. The residue was extracted with dichloromethane (2 X 15 10 mL). The combined dichloromethane t was concentrated under reduced pressure and the residue was purified with reverse-phase prep-HPLC to afford 159 (28 mg, 22%). MS-ESI: [M+H]+ 681. 1H NMR (500 MHz, CDCl3) δ 8.71 (d, J = 2.5 Hz, 1H), 8.65 (d, J = 5.0 Hz, 1H), 8.30 (s, 1H), 8.04 (s, 1H), 7.87 (s, 1H), 7.70 (d, J = 2.0 Hz, 1H), 7.55 (d, J = 5.0 Hz, 1H), 7.37 (d, J = 9.0 Hz, 1H), 6.81 (d, J = 9.0 Hz, 1H), 4.67-4.61 (m, 2H), 4.71-4.64 (m, 2H), 20 4.44-4.42 (m, 2H), 4.34-4.33 (m, 1H), 3.83-3.76 (m, 1H), 3.72 (s, 3H), 3.20-3.16 (m, 1H), 2.99-2.84 (m, 6H), 2.79-2.71 (m, 2H), 2.50-2.48 (m, 1H), 2.02-1.98 (m, 4H), 0.91 (d, J = 6.0 Hz, 6H).
Example 160a 4-(5-(5-((2S,5R)-2,5-Dimethyl(oxetanyl)piperazin yl)pyridinylamino) methyloxo-1,6-dihydropyridinyl)(1-oxo-6,7,8,9- 25 ydropyrazino[1,2-a] 2(1H)-yl)nicotinaldehyde 160a A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 4-chloro(1-oxo-6,7,8,9-tetrahydropyrazino[1,2-a]indol- 206 2(1H)-yl)nicotinaldehyde 103b (150 mg, 1.0 eq., 0.46 mmol), 3-(5-((2S,5R)-2,5-dimethyl (oxetanyl)piperazinyl)pyridinylamino)methyl(4,4,5,5-tetramethyl-1,3,2- dioxaborolanyl)pyridin-2(1H)-one 155a (228 mg, 0.46 mmol), K3PO4 (195 mg, 0.92 mmol), PdCl2(dppf) (37 mg, 0.046 mmol), sodium acetate (75 mg, 0.92 mmol), water (8 5 drops), and acetonitrile (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at 80ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under d pressure and the resulting residue was purified by silica-gel column chromatography eluting with 40:1 dichloromethane/ethanol to afford 160a as yellow solid (80 mg, 26%). MS-ESI: [M+H]+ 661.4. 10 Example 160 2-{5-[5-((2S,5R)-2,5-Dimethyloxetanyl-piperazinyl)-pyridin- 2-ylamino]-3'-hydroxymethylmethyloxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}-6,7,8,9- tetrahydro-2H-pyrazino[1,2-a]indolone 160 A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with 160a (80 mg, 0.12 mmol), NaBH4 (23 mg, 0.60 mmol), and methanol (10 mL). 15 The mixture was stirred at room temperature for 1 h. It was quenched with water (1 mL) and filtered. The te was concentrated under reduced pressure and the e was purified by reverse-phase prep-HPLC to afford 160 (44 mg, 55%). MS-ESI: [M+H]+ 663.3. 1H NMR (500 MHz, CDCl3) δ 8.72-8.70 (m, 1H), 8.57 (d, J = 5.0 Hz, 1H), 8.03 (d, J = 2.5 Hz, 1H), 7.89-7.87 (m, 2H), 7.50 (d, J = 5.0 Hz, 1H), 7.37-7.35 (m, 1H), 7.06 (s, 1H), 6.97 (d, J = 6.0 20 Hz, 1H), 6.81 (d, J = 9.0 Hz, 1H), 6.68 (d, J = 6.0 Hz, 1H), 5.10-5.08 (m, 1H), 4.76-4.32 (m, 6H), 3.76-3.72 (m, 4H), 3.20-3.17 (m, 1H), 2.93-2.90 (m, 1H), 2.76-2.69 (m, 6H), .46 (m, 1H), 1.97-1.94 (m, 3H), 1.87-1.84 (m, 2H), 0.89 (t, J = 6.5 Hz, 6H).
Example 161a rt-Butyl 3-Ethyl(6-nitropyridinyl)piperazine carboxylate 161a 25 A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with oxane (50 mL), 5-bromonitropyridine (2.02 g, 10 mmol), (S)-tert-butyl 3-ethylpiperazinecarboxylate (2.14 g, 10.0 mmol), Pd2(dba)3 (458 mg, 0.50mmol), XantPhos (576 mg, 1.0 mmol), and cesium carbonate (6.52 g, 20 mmol). 30 After three cycles of vacuum/argon flush, the e was heated at 100oC overnight. After 207 this time the on was cooled to room temperature. It was then filtered and the filtrate was evaporated under reduced pressure. The residue was purified by silica-gel column tography eluting with 3:1 petroleum ethyl acetate to afford 161a (700 mg, 22%) as a yellow solid. MS: [M+H]+ 336 5 Example 161b (S)-tert-Butyl minopyridinyl)ethylpiperazine carboxylate 161b A 100-mL single-neck round-bottomed flask was purged with nitrogen and charged with 161a (0.7 g, 2.08 mmol), 10% palladium on carbon (50% wet, 208 mg), and methanol (40 mL). The mixture was evacuated, charged with hydrogen gas, and stirred at room 10 temperature for 6 h. The hydrogen was then ted and en was charged into the flask. The catalyst was removed by filtration through a pad of CELITE® and the filtrate was concentrated under reduced pressure to afford 161b (568 mg, 89%). MS: [M+H]+ 306 Example 161c (S)-tert-Butyl 4-(6-(5-Bromomethyloxo-1,2- dihydropyridinylamino) pyridinyl)ethylpiperazinecarboxylate 161c 15 A 100-mL single-neck bottomed flask equipped with a magnetic r and a reflux condenser was charged with 1,4-dioxane (50 mL), 161b (568 mg, 1.86 mmol), 3,5- dibromomethylpyridin-2(1H)-one (498 mg, 1.86 mmol), Pd2(dba)3 (85 mg, 0.093mmol), XantPhos (107 mg, 0.186 mmol), and cesium carbonate (1.198 g, 3.72 mmol). After three cycles of vacuum/argon flush, the mixture was heated at 100 oC for 6 h. It was then filtered 20 and the filtrate was evaporated under reduced pressure. The e was purified by silica-gel column chromatography eluting with 100:1 dichloromethane/methanol to afford 161c (502 mg, 55%) as a yellow solid. MS: [M+H]+ 492.
Example 161d (S)Bromo(5-(2-ethylpiperazinyl)pyridinylamino) methyl pyridin-2(1H)-one 161d 25 A mixture of 161c (502 mg, 1.02 mmol), dichloromethane (2 mL), and 4.0 M HCl/dioxane (4 mL) was stirred at room temperature for 5 h. It was then concentrated under reduced pressure to afford crude 161d as a yellow solid (263 mg, 66%), which was used in the next step without further purification. MS: [M+H]+ 392.
Example 161e (S)Bromo(5-(2-ethyl(oxetanyl)piperazin 30 yl)pyridinylamino) methylpyridin-2(1H)-one 161e 208 A mixture of 161d (263 mg, 0.67 mmol), oxetanone (96 mg, 1.34 mmol), NaBH3CN 104 mg, 1.68 mmol), and zinc chloride (227 mg, 1.68 mmol) in methanol (10 mL) was stirred at 50ºC for 5 hours. Water (10 mL) was then added to the reaction. The resulting 5 mixture was concentrated under reduced pressure. The residue was extracted with dichloromethane three times. The combined organic layer was concentrated under reduced pressure and the residue was purified by silica-gel column tography eluting with 50:1 dichloromethane/methanol to afford 161e (203 mg, 68%). MS: [M+H]+ 448.
Example 161f (S)(5-(2-Ethyl(oxetanyl)piperazinyl)pyridin 10 ylamino)methyl (4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 161f A 100-mL single-neck round-bottomed flask ed with a ic r and a reflux condenser was charged with 161e (3219 mg, 7.20 mmol), Pin2B2 (9072 mg, 36.0 mmol), Pd2(dba)3 (329 mg, 0.36 mmol), X-phos (302 mg, 0.72 mmol), potassium e (2117 mg, 21.6mmol), and dioxane (50 mL). After three cycles of vacuum/argon flush, the 15 mixture was heated at 60 ºC for 16 h. It was then cooled to room temperature and filtered.
The filtrate was concentrated under reduced pressure and the resulting residue was washed with 8:1 petroleum ether/ethyl acetate (80 mL) to afford 161f as yellow solid (3.0 g, 84%).
MS: [M+H]+ 496.4.
Example 161g 4-(5-(5-(2-Ethyl(oxetanyl)piperazinyl)pyridin 20 ylamino)methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- hexahydropyrido[3,4-b]indolizin-2(1H)-yl)nicotinaldehyde 161g A 50-mL round-bottomed flask equipped with a reflux condenser was charged with 161f (200 mg, 0.40 mmol), 4-chloro(1-oxo-3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin- 25 2(1H)-yl)nicotinaldehyde 139a (132 mg, 0.40 mmol), K3PO4 3water (213 mg, 0.80 mmol), 209 sodium acetate (66 mg, 0.80 mmol), 1,1’- bis(diphenylphosphino)ferrocenedichloropalladium(II) (16 mg, 0.020 mmol), and acetonitrile (20 mL). After three cycles of vacuum/N2 flush, the mixture was heated at 100°C under N2 protection for 2 h. Analysis of the on mixture by LCMS showed complete conversion to 5 the desired product. The on mixture was cooled to room ature and filtered. The filtrate was concentrated under reduced re. The residue was diluted with dichloromethane (50 mL) and water (50 mL). The aqueous layer was separated and extracted with dichloromethane (3 × 20 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The dark residue was purified by silica-gel 10 column chromatography eluting with dichloromethane/methanol (80/1 to 30/1) to afford 161g (150 mg, 57%) as yellow solid. MS-ESI: [M+H]+ 663 Example 161 2-{5-[5-((S)Ethyloxetanyl-piperazinyl)-pyridin ylamino]-3'-hydroxymethylmethyloxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}-2,3,5,6,7,8- hexahydro-4H-2,4b-diaza-fluorenone 161 15 To a solution of 161g (120 mg, 0.18 mmol) in methanol (20 mL) was added NaBH4 (21 mg, 0.54 mmol) at room temperature. After the reaction was d for 1 h, LCMS ted the reaction was complete. Then the mixture was poured into water (20 mL) and concentrated under reduced pressure. The residue was extracted with dichloromethane (3 X 40 mL). The combined c layer was washed with brine (20 mL), dried over Na2SO4, 20 filtered, and concentrated. The e solid was purified by prep-HPLC to afford 161 (97 mg, 81%) as white solid. MS-ESI: [M+H]+ 665. 1H NMR (500 MHz, CDCl3) δ 8.63 (s, 1H), 8.50 (d, J = 5.0 Hz, 1H), 7.93 (s, 1H), 7.84-7.82 (m, 2H), 7.35 (d, J = 4.5 Hz, 1H), 7.28 (s, 1H), 6.82 (d, J = 9.0 Hz, 1H), 6.32 (s, 1H), 5.01-4.99 (m, 1H), 4.73-4.64 (m, 5H), 4.45-4.40 (m, 1H), 4.30 (t, J = 12.0 Hz, 1H), 3.94-3.91 (m, 1H), 3.85-3.83 (m, 2H), 3.72 (s, 3H), 3.55-3.53 25 (m, 1H), 3.34-3.32 (m, 1H), 3.14-3.12 (m, 2H), 3.04-2.92 (m, 2H), 2.84-2.82 (m, 2H), 2.59- 2.57 (m, 1H), 2.46-2.44 (m, 2H), 2.38-2.36 (m, 1H), 2.06-2.01 (m, 2H), 1.90-1.86 (m, 2H), 1.68-1.66 (m, 1H), 1.43-1.39 (m, 1H), 0.82 (t, J = 7.5 Hz, 3H).
Example 162a 5-Bromomethyl(pyrazinylamino)pyridin-2(1H)-one 162a 30 A 100-mL single-neck round-bottomed flask equipped with a ic stirrer and a reflux condenser was charged with pyrazinamine (500 mg, 5.3 mmol), 3,5-dibromo methyl pyridin-2(1H)-one (1335 mg, 5.3 mmol), Pd2(dba)3 (229 mg, 0.25 mmol), XantPhos (289 mg, 0.50 mmol), cesium carbonate (3.26 g, 10 mmol) and 1,4-dioxane (50 mL). After three cycles of vacuum/argon flush, the mixture was stirred at 100oC for 2 h. It was then 210 filtered and the filtrate was evaporated in vacuo. The residue was purified by silica-gel column chromatography eluting with 50:1 dichloromethane/methanol to afford 162a (420 mg, 30%) as a yellow solid. MS-ESI: [M+H]+ 281.0.
Example 162b (4-(1-Methyloxo(pyrazinylamino)-1,6-dihydropyridin- 5 3-yl)(1-oxo- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl acetate 162b A 100-mL single-neck round-bottomed flask equipped with a reflux condenser was charged with 162a (170 mg, 0.61 mmol), 3-(acetoxymethyl)(1-oxo-3,4,6,7,8,9- 10 hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinylboronic acid 113i (280 mg, 0.72 mmol), Pd(dppf)Cl2 (30 mg, 0.037 mmol), K3PO4·trihydrate (270 mg, 1.2 mmol), sodium acetate (180 mg, 1.2 mmol), acetonitrile (20 mL), and water (0.5 mL). The system was evacuated and refilled with N2. The reaction mixture was stirred at 100 ºC for 2 h. It was then cooled to room temperature and ed. The filtrate was concentrated under reduced pressure and the 15 resulting residue was purified by silica-gel column chromatography eluting with 25:1 of dichloromethane/methanol to afford 162b (130 mg, 40%) as a yellow brown solid. : [M+H]+ 540.3 Example 162 2-[3'-Hydroxymethylmethyloxo(pyrazinylamino)-1,6- dihydro-[3,4']bipyridinyl-2'-yl]-3,4,6,7,8,9-hexahydro-2H-pyrazino[1,2-a]indolone 162 20 A mixture of 162b (110 mg, 0.20 mmol) and lithium hydroxide (84 mg, 2.0 mmol) in anol/THF (1:1, 4 mL) and water (1 mL) was d at 30oC for 1 h. The mixture was evaporated in vacuo and the residue was d with water and ethyl acetate. The water phase was separated and extracted with ethyl acetate (2 x 10 mL). The combined ethyl e extract was trated under reduced pressure and the residue was purified by reverse- 25 phase prep-HPLC to afford 162 (85 mg, 85%) as pale yellow solid. MS-ESI: [M+H]+ 498.3. 1H NMR (500 MHz, CHCl δ 8.73 (d, J = 2.5 Hz, 1H), 8.54 (d, J = 5 Hz, 1H), 8.29 (s, 1H), 3) 8.15-8.14 (m, 2H), 8.01 (d, J = 2.5 Hz, 1H), 8.00 (d, J = 2.0 Hz, 1H), 7.37(d, J = 5 Hz, 1H), 6.91 (s, 1H), 4.66-4.65 (m, 1H), 4.52-4.51 (m, 1H), 4.32-4.31 (m, 1H), 4.18-4.17 (m, 1H), 211 4.14-4.12 (m, 1H), .88 (m, 1H), 3.75 (s, 3H), 2.62-2.57 (m, 4H), 1.92-1.88 (m, 3H), 1.80-1.79 (m, 2H). e 163a 1-Methyl(5-(oxetanyl)-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinylamino)oxo-1,6-dihydropyridinylboronic acid 163a 5 A 100-mL round bottomed flask equipped with a magnetic stirrer was charged with 5- bromomethyl(5-(oxetanyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin ylamino)pyridin-2(1H)-one 125i (1.0 g, 2.64 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2' - bi(1,3,2-dioxaborolane) (2.0 g, 7.92 mmol), PdCl2(dppf) (190 mg, 0.26 mmol), potassium acetate (776 mg, 7.92 mmol), and e (40 mL). After bubbling argon into the mixture for 10 30 minutes, a reflux condenser was attached to the flask and e was stirred at 100oC for 6 h under an argon atmosphere. The resulting mixture was filtered and the filtrate was evaporated under reduced pressure. The residue was purified by reverse-phase Combiflash eluting with 0.3% NH4HCO3 water/CH3CN to afford 163a as a white solid (300 mg, 33%).MS: [M+H]+ 346. 15 Example 163b 2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}(1-methyl{[5-(oxetanyl)-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin no}oxo-1,6-dihydropyridinyl)pyridinecarbaldehyde 163b O N N N NH O O N N N O N 163b A 100-mL single-neck round-bottomed flask equipped with a reflux condenser was 20 charged with (4-chloro{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- dienyl}pyridinecarbaldehyde) 108a (280 mg, 0.81 mmol), 163a (440 mg, 0.96 mmol), Pd(dppf)Cl2 (40 mg, 0.049 mmol), K3PO4 (360 mg, 1.6 mmol), sodium acetate trihydrate(240 mg, 1.6 mmol), water (6 drops), and acetonitrile (20 mL). The system was evacuated and ed with N2. The reaction mixture was stirred at 100 ºC for 2 h. It was then cooled to 25 room temperature and filtered. The filtrate was concentrated under d pressure and the resulting residue was purified by silica-gel column chromatography eluting with 25:1 dichloromethane/methanol to afford 163b (150 mg, 31%) as a yellow brown solid. MS-ESI: [M+H]+ 609.3 212 Example 163 2-[3'-Hydroxymethylmethyl(5-oxetanyl-4,5,6,7-tetrahydropyrazolo [1,5-a]pyrazinylamino)oxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl]-7,7-dimethyl- 3,4,7,8-tetrahydro-2H,6H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazinone 163 A mixture of 163b (80 mg, 0.12 mmol) and NaBH4 (15 mg, 0.36 mmol) in ol 5 (5 mL) was stirred at 30oC for 2 h. The e was quenched with water and concentrated under reduced pressure. The residue was extracted with ethyl acetate (3 X 10 mL). The ed ethyl acetate extract was concentrated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 163 (30 mg, 50%) as dark red solid. MS-ESI: [M+H]+ 611.4. 1H NMR (500 MHz, CHCl3) δ 8.48 (d, J = 5.5 Hz, 1H), 7.96 (d, J = 2.5 Hz, 10 1H), 7.70 (d, J = 2, 1H), 7.43 (s, 1H), 7.34 (d, J = 5 Hz, 1H), 6.8 (s, 1H), 5.70 (s, 1H), 5.03 (t, J = 6, 1H), 4.77-4.73 (m, 3H), 4.68 (t, J = 6.5 Hz, 2H), 4.51-4.50 (m, 1H), 4.34-4.33 (m, 1H), 4.23-4.16 (m, 2H) , 4.09 (t, J = 5.5 Hz, 2H), 3.86-3.85 (m, 1H), 3.79-3.74 (m, 1H), 3.71 (s, 3H ), 3.56 (d, J = 4, 2H), 2.83 (t, J = 5.5 Hz, 2H), 2.58 (d, J = 5.5 Hz, 2H), 2.52 (s, 2H ), 1.28 (s, 6H ). 15 Example 164a 2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}[5-({5-[(2S)ethyl(oxetanyl)piperazinyl]pyridinyl}amino)- 1-methyloxo-1,6-dihydropyridinyl]pyridinecarbaldehyde 164a O N N N NH O O N N N O N 164a A 100-mL -neck round-bottomed flask equipped with a reflux condenser was 20 charged with (4-chloro{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- dienyl}pyridinecarbaldehyde) 108a (280 mg, 0.8 mmol), (S)(5-(2-ethyl(oxetan- 3-yl)piperazinyl)pyridinylamino)methyl(4,4,5,5-tetra-methyl-1,3,2-dioxaborolan- 2-yl)pyridin-2(1H)-one 161f (500 mg, 0.96 mmol), Pd(dppf)Cl2 (33 mg, 0.040 mmol), K3PO4 (360 mg, 1.6 mmol), sodium acetate trihydrate (240 mg, 1.6 mmol), and acetonitrile (100 25 mL). The system was evacuated and refilled with N2. The reaction mixture was heated at 100ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel 213 column chromatography eluting with 25:1 dichloromethane/methanol to afford 164a (320 mg, 60%) as a yellow brown solid. MS-ESI: [M+H]+ 677.3.
Example 164 5-((S)Ethyloxetanyl-piperazinyl)-pyridin ylamino]-3'-hydroxymethylmethyloxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}-7,7- 5 dimethyl-3,4,7,8-tetrahydro-2H,6H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazinone 164 A mixture of 164a (200 mg, 0.30 mmol) and NaBH4 (36 mg, 0.90 mmol) in methanol (30 mL) was stirred at 30oC for 2 h. The e was quenched with water and concentrated under reduced pressure. The residue was extracted with ethyl acetate (3 X 10 mL). The combined ethyl acetate extract was concentrated under reduced pressure and the residue was 10 purified by e-phase prep-HPLC to afford 164 (140 mg, 72%) as light green solid. MSESI : [M+H]+ 679.3. 1H NMR (500 MHz, CDCl δ 8.63 (d, J = 2.5 Hz, 1H), 8.49 (d, J = 5.0 3) Hz, 1H), 7.93 (d, J = 2.5 Hz, 1H), 7.83 (s, 1H), 7.82 (s, 1H), 7.37 (d, J = 5.0 Hz, 1H), 7.27 (s, 1H), 6.85 (s, 1H), 6.82 (d, J = 9.0 Hz, 1H), 5.06 (s, 1H), 4.71-4.61 (m, 5H), .50 (m, 1H), 4.34-4.32 (m, 1H), 4.16 (d, J = 4.5 Hz, 2H), 3.87-3.85 (m, 1H), 3.72 (s, 3H), 3.55-3.50 15 (m, 1H), 3.33-3.30 (m, 1H), 3.12 (t, J = 5.0 Hz, 2H), 2.58-2.55 (m, 3H), 2.52 (s, 2H), 2.44 (d, J = 3.5 Hz, 2H), 2.35 (t, J = 5.5 Hz, 1H), 1.68-1.64 (m, 1H), .37 (m, 1H), 1.28 (s, 6H), 0.82 (t, J = 7.5 Hz, 3H).
Example 165a (S)(5-(5-(2-Ethyl(oxetanyl)piperazinyl)pyridin ylamino)methyloxo-1,6-dihydropyridinyl)(1-oxo-6,7,8,9-tetrahydropyrazino[1,2- 20 a]indol-2(1H)-yl)nicotinaldehyde 165a A 50-mL flask equipped with a reflux condenser was charged with 4-chloro(1-oxo- 6,7,8,9-tetrahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 103b (164 mg, 0.50 mmol), (S)(5-(2-ethyl(oxetanyl)piperazinyl)pyridinylamino)methyl(4,4,5,5- 25 tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 161f (347 mg, 0.70 mmol), potassium acetate (137 mg, 1.4 mmol), 1,1’-bis(diphenylphosphino)ferrocenedichloropalladium(II) (29 mg, 0.035 mmol), water (5 drops), and acetonitrile (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100 °C under argon atmosphere for 3 h. The 214 reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue was d with dichloromethane (50 mL) and water (50 mL). The aqueous layer was separated and ted with dichloromethane (3 × 20 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under d 5 re. The dark residue was purified by silica-gel column chromatography eluting with romethane / methanol (80/1 to 25/1) to afford 165a (151 mg, 46%) as yellow solid.
MS-ESI: [M+H]+ 661 Example 165 2-{5-[5-((S)Ethyloxetanyl-piperazinyl)-pyridin ylamino]-3'-hydroxymethylmethyloxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}-6,7,8,9- 10 tetrahydro-2H-pyrazino[1,2-a]indolone 165 To a solution of 165a (100 mg, 0.15 mmol) in methanol (10 mL) was added NaBH4 (34 mg, 0.90 mmol) at room temperature. After the reaction was stirred for 1h, LCMS indicated the reaction was te. Then the mixture was quenched with water (8 mL) and concentrated under reduced pressure. The residue was extracted with dichloromethane (3 X 15 10 mL). The combined organic layer was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The e was purified by reverse-phase prep-HPLC to afford 165 (35 mg, 35%) as light yellow solid. MS-ESI: [M+H]+ 663. 1H NMR (500 MHz, DMSO-d6) δ 8.64 (s, 1H), 8.57 (d, J = 5.0 Hz, 1H), 8.44 (s, 1H), 7.82 (d, J = 2.0 Hz, 1H), 7.51-7.48 (m, 2H), 7.35 (dd, J = 2.0 Hz, 9.0 Hz, 1H), 7.27-7.23 (m, 2H), 6.83-6.81 20 (m, 2H), 4.97 (bs, 1H), 4.59-4.55 (m, 2H), 4.49-4.32 (m, 4H), 3.61 (s, 3H), 3.51 -3.47 (m, 1H), 3.42-3.37 (m, 1H), .16 (m, 1H), 3.01-2.98 (m, 1H), 2.76-2.74 (m, 2H), 2.63-2.61 (m, 3H), 2.55-2.54 (m, 1H), 2.19-2.16 (m, 1H), 2.12-2.07 (m, 1H), 1.90-1.85 (m, 2H), 1.77- 1.66 (m, 3H), 1.27-1.25 (m, 1H), 0.79 (t, J = 7.0 Hz, 3H).
Example 166a 2-(10-Fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 25 2(1H)-yl)(1-methyl(5-(oxetanyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin ylamino)oxo-1,6-dihydropyridinyl)nicotinaldehyde 166a 215 A 100-mL single-neck round-bottomed flask ed with a magnetic stirrer and reflux condenser was charged with1-methyl(5-(oxetanyl)-4,5,6,7- tetrahydropyrazolo[1,5-a] pyrazinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolan yl)pyridin-2(1H)-one 163a (354 mg, 0.83 mmol), 4-chloro(10-fluorooxo-3,4,6,7,8,9- 5 hexahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 134c (289 mg, 0.83 mmol), PdCl2(dppf) (68 mg, 0.08 mmol), K3PO4 (352 mg, 1.66 mmol), sodium acetate (136 mg, 1.66 mmol), acetonitrile (50 mL), and water (3 mL). The system was evacuated and refilled with N2. The reaction mixture was heated at 100 ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the 10 resulting residue was purified by silica-gel column chromatography g with 30:1 romethane/methanol to afford 166a (305 mg, 60%) as a brown solid. MS-ESI: [M+H]+:613.6.
Example 166 10-Fluoro[3'-hydroxymethylmethyl(5-oxetanyl-4,5,6,7- tetrahydro-pyrazolo[1,5-a]pyrazinylamino)oxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl]- 15 3,4,6,7,8,9-hexahydro-2H-pyrazino[1,2-a]indolone 166 To a suspension of 166a (250 mg, 0.41 mmol) in methanol (20 mL) was added sodium borohydride (47 mg, 1.23 mmol) at 0 oC. The e was stirred for 30 minutes. It was then quenched with water (2 mL) and concentrated under reduced pressure. The residue was purified by reverse-phase prep-HPLC to afford 166 (20 mg, 6.6 %). MS-ESI: [M+H]+ 20 615.6. 1H NMR (500 MHz, CDCl3) δ 8.46 (d, J = 5.0 Hz, 1H), 7.94 (d, J = 3.0 Hz, 1H), 7.66 (d, J = 2.0 Hz, 1H), 7.43 (s, 1H), 7.34 (d, J = 5.0 Hz, 1H), 5.75 (s, 1H), 4.95 (t, J = 6.5 Hz, 1H), 4.76-4.74 (m, 2H), 4.69-4.65-4.67 (m, 3H), 4.46-4.44 (m, 1H), .33 (m, 1H), 4.10- 4.08 (m, 4H), .35 (m, 2H), 3.69 (s, 3H), .56 (m, 2H), 2.842.82 (m, 2H), 2.58- 2.53 (m, 4H), 1.89-1.84 (m, 2H), 1.77-1.76 (m, 2H). 25 Example 167a robicyclo[2.2.1]heptenecarbaldehyde 167a A 250-mL three-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was purged with nitrogen and charged with anhydrous 1,2-dichloroethane (24 mL) and anhydrous DMF (9.12 g, 125 mmol). The reaction mixture was cooled to 0oC 30 and phosphorus oxychloride (15.3 g, 100 mmol) was added over a period of 5 minutes while maintaining the reaction temperature between 0 and 10 oC. The cooling bath was removed and the on was stirred at room temperature for 30 minutes. A solution of 216 bicyclo[2.2.1]heptanone (5.50 g, 50.0 mmol) in 1,2-dichloroethane (10 mL) was added and the resulting mixture was heated at 80ºC overnight. After this time, the on was poured into a solution of potassium monohydrogen phosphate (43.5 g, 250 mmol) in water (200 mL) and stirred for 15 minutes. The organic layer was separated and concentrated under 5 reduced pressure. The residue was dissolved in methylene chloride (300 mL) and washed with water (2 x 50 mL). The methylene chloride layer was dried over sodium sulfate, filtered, and concentrated under d pressure. The residue was purified by silica-gel column chromatography eluting with 1:100 ethyl e/petroleum ether to afford 167a as a yellow oil (2.2 g, 28%). MS: [M+H]+ 157. 1H NMR (500 MHz, CDCl3) δ 9.80 (s, 1H), 3.42-3.41 (m, 10 1H), 3.08-3.07 (m, 1H), 1.95-1.77 (m, 2H), 1.68-1.66 (m, 1H), .17 (m, 3H).
Example 167b (E)-Ethyl 3-(3-Chlorobicyclo[2.2.1]heptenyl)acrylate 167b To a solution of 167a (9.0 g, 57.7 mmol) in methylene chloride (250 mL) was added 15 ethyl 2-(triphenyl-λ5-phosphanylidene)acetate (20 g, 57.7 mmol). The mixture was stirred at room temperature overnight. It was then ated under reduced pressure. The residue was purified by -gel column chromatography g with 1:100 ethyl acetate/petroleum ether to afford 167b as a yellow oil (6.0 g, 46%). MS: [M+H]+ 227.
Example 167c Ethyl 3-Azatricyclo[5.2.1.02,6]deca-2(6),4-dienecarboxylate 20 167c To a solution of 167b (5.0 g, 22 mmol) in DMSO (30 mL) was added NaN3 (2.2 g, 33 mmol). The mixture was heated at 105ºC for 6 hours. Water (13 mL) was added to the reaction mixture after cooling down to room temperature and the resulting mixture was 25 extracted with methylene chloride (3 X 50 mL). The combined organic phase was dried over Na2SO4 and evaporated under reduced pressure to dryness. The residue was ed by silica-gel column chromatography eluting with 20:1 methylene chloride/methanol to afford 167c as a brown solid (2.7 g, 60%). MS: [M+H]+ 206. 1H NMR (500 MHz, CDCl3) δ 11.51 217 (s, 1H), 6.45 (s, 1H), 4.16 (q, J = 6.5 Hz, 2H), 3.26-3.24 (m, 2H), 1.82-1.79 (m, 2H), 1.74- 1.72 (m, 2H), 1.24 (t, J = 6.5 Hz, 3H), 0.91-0.89 (m, 2H).
Example 167d Ethyl 3-(Cyanomethyl)azatricyclo[5.2.1.02,6]deca-2(6),4- diene carboxylate 167d N N O O 5 Into a on of 167c (3.0 g, 14.6 mmol) in anhydrous DMF (30 mL) was added NaH (880 mg, 22 mmol). The mixture was stirred at room temperature for 30 minutes. 2- Bromoacetonitrile (3.5 g, 29.3 mmol) was added and the resulting mixture was heated at 65ºC for 1 hour. It was then stirred at room temperature overnight. After reaction water (30 mL) 10 was added and the resulting mixture was extracted with ethyl acetate (200 mL × 3). The combined organic phase was evaporated under d pressure to dryness. The e was purified by -gel column chromatography eluting with 20:1 methylene chloride/methanol to afford 167d as a brown solid (2.6 g, 72%). MS: [M+H]+ 245.
Example 167e Ethyl 3-(2-Aminoethyl)azatricyclo[5.2.1.02,6]deca-2(6),4- 15 diene ylate 167e A suspension of 167d (4.0 g, 16 mmol) and Raney Ni (400 mg) in methanol (60 mL) was hydrogenated in a Parr apparatus at 50 psi overnight. The mixture was filtered through a pad of ® and the filtrate was concentrated under reduced pressure. The residue was 20 purified by silica-gel column chromatography eluting with 20:1 methylene chloride/methanol to afford 167e as a yellow solid (2.0 g, 50%). MS: [M+H]+ 249.
Example 167f 3,6-Diazatetracyclo[9.2.1.02,10.03,8]tetradeca-2(10),8-dienone 167f N NH O 218 Into a solution of 167e (1.8 g, 7.2 mmol) in ethanol (40 mL) was added sodium methoxide (2.5 g, 36 mmol). The mixture was heated at 65ºC for 12 hours. It was then cooled to room temperature. The t was evaporated to dryness under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 20:1 ene 5 chloride/methanol to afford the racemate as a brown solid (800 mg, 53%), chiral resolution of which ed 167f and 170a. MS: [M+H]+ 203. e 167g 4-Chloro[(1S,11R)oxo-3,6- diazatetracyclo[9.2.1.02,10.03,8]tetradeca-2(10),8-dienyl]pyridinecarbaldehyde 167g A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 10 reflux condenser was charged with oxane (30 mL), 167f (400 mg, 2.0 mmol), 2-bromo- 4-chloronicotinaldehyde (1.30 g, 6.0 mmol), and potassium acetate (390 mg, 4.0 mmol).
After bubbling nitrogen through the resulting mixture for 30 minutes, Xantphos (110 mg, 0.20 mmol) and tris(dibenzylideneacetone)dipalladium(0) (180 mg, 0.20 mmol) were added and the reaction mixture was heated at 80ºC for 10 h. It was then cooled to room temperature 15 and filtered. The filtrate was partitioned between ethyl acetate (50 mL) and water (30 mL).
The s layer was separated and extracted with ethyl acetate (3 X 30 mL). The combined organic layer was washed with brine (20 mL) and dried over sodium sulfate. The drying agent was removed by filtration and the filtrate was concentrated under reduced pressure. The residue was purified with silica-gel column chromatography eluting with 2:1 petroleum 20 ether/ethyl acetate to afford 167g (391 mg, 57%) as yellow solid. MS-ESI: [M+H]+ 342.2 e 167h 4-[1-Methyl({5-methyl-4H,5H,6H,7H-pyrazolo[1,5- zinyl}amino)oxo-1,6-dihydropyridinyl][(1S,11R)oxo-3,6- diazatetracyclo[9.2.1.02,10.03,8]tetradeca-2(10),8-dienyl]pyridinecarbaldehyde 167h N N N NH O N O O N N Cl N N 135a O N O PdCl2(dppf), K3PO4, H2O N AcONa, CH3CN, 100 °C, 3h 167g 167h 25 A 100-mL round-bottomed flask equipped with a reflux condenser was charged with 167g (150 mg, 0.44 mmol), 1-methyl(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin- 2-ylamino)(4,4,5,5-tetra-methyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 135a (169 mg, 219 0.44 mmol), sodium acetate (72 mg, 0.88 mmol), K3PO4 (234 mg, 0.88 mmol), PdCl2(dppf) (36 mg, 0.044 mmol), acetonitrile (20 mL), and water (1 mL). After ng nitrogen through the mixture for 30 s, it was heated at 100ºC under N2 for 3 hours. The reaction mixture was cooled to room temperature and filtered. The filtrate was trated under 5 reduced pressure and the residue was ed by silica-gel column chromatography eluting with 25:1 dichloromethane/methanol to afford 167h (132 mg, 53%) as a brown solid. MSESI : [M+H]+ 565.3 Example 167 2-(3-(hydroxymethyl)(1-methyl(5-methyl-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 10 3,4,6,7,8,9-hexahydro-6,9-methanopyrazino[1,2-a]indol-1(2H)-one 167 A solution of 167h (120 mg, 0.21 mmol) in methanol (20 mL) was added NaBH4 (24 mg, 0.63 mmol). The e was stirred at 20ºC for 2 h. The reaction was quenched with water and concentrated under reduced pressure. The residue was purified by reverse-phase prep-HPLC to afford 167 (98 mg, 83%) as a yellow solid. MS-ESI: [M+H]+ 567.3. 1H NMR 15 (500 MHz, DMSO-d6) δ 8.48 (d, J = 5.0 Hz, 1H), 8.19 (s, 1H), 8.05 (d, J = 2.5 Hz, 1H), 7.40 (s, 1H), 7.32 (d, J = 5.0 Hz, 1H), 6.53 (d, J = 5.5 Hz, 1H), 5.89 (s, 1H), 4.98 (t, J = 5.0 Hz, 1H), 4.48-4.30 (m, 3H), 4.27-4.22 (m, 2H), 3.92-3.91 (m, 2H), 3.86-3.84 (m, 1H), 3.60 (s, 3H), 3.52-3.33 (m, 3H), 3.29 (ps, 1H), 2.79-2.77 (m, 2H), 2.35 (s, 3H), 1.87-1.76 (m, 3H), 1.60-1.59 (m, 1H), .91 (m, 2H). 20 Example 168a 3-(6,7-Dihydro-4H-pyrazolo[5,1-c][1,4]oxazinylamino) methyl(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 168a A 250-mL round bottomed flask equipped with a magnetic stirrer and a reflux condenser was d with the mixture of 5-bromo(6,7-dihydro-4H-pyrazolo[5,1- c][1,4]oxazinylamino)methylpyridin-2(1H)-one 129c (1.3 g, 4.0 mmol), 25 bis(pinacolato)diboron (2.03 g, 8.0 mmol), PdCl2(dppf) (439 mg, 0.60 mmol), potassium acetate (784 mg, 8.0 mmol), and 1,4-dioxane (60 mL). After bubbling nitrogen through the mixture for 30 minutes, it was heated at reflux for 15 h. The mixture was cooled to room ature upon completion of the reaction and filtered. The solid was washed with ethyl acetate (100 mL). The combined filtrate was evaporated under d pressure and the 30 residue was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 168a (446 mg, 30%). MS: [M+H]+ 373.
Example 168b 4-(1-Methyl(6,7-dihydro-4H-pyrazolo[5,1- c][1,4]oxazinylamino)oxo- 1,6-dihydropyridinyl)(4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl)nicotinaldehyde 168b 220 A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with ro{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 108a (200 mg, 5 0.58 mmol), 168a (433 mg, 1.16 mmol), PdCl2(dppf) (48 mg, 0.052 mmol), K3PO4 (246 mg, 1.16 mmol), sodium acetate (96 mg, 1.16 mmol), acetonitrile (10 mL), and water (0.5 mL).
The system was evacuated and refilled with N2. The reaction mixture was stirred at 100ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography 10 eluting with 30:1 dichloromethane/methanol to afford 168b (250 mg, 78%) as a yellow solid.
MS-ESI: [M+H]+ 554.6. e 168 2-[5-(6,7-Dihydro-4H-pyrazolo[5,1-c][1,4]oxazinylamino)-3'- ymethylmethyloxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl]-7,7-dimethyl-3,4,7,8- tetrahydro-2H,6H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazinone 168 15 To a suspension of 168b (200 mg, 0.36 mmol) at 0oC in methanol (10 mL) was added sodium borohydride (42 mg, 1.1 mmol). The reaction mixture was stirred for 30 minutes and ed with water (2 mL). It was then concentrated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 168 (53 mg, 21 %) as a yellow solid. MSESI : [M+H]+ 556.6. 1H NMR (400 MHz, CDCl3) δ 8.46 (d, J = 5.2 Hz, 1H), 7.97 (d, J = 2.4 20 Hz, 1H), 7.71 (d, J = 2.0 Hz, 1H), 7.45 (s, 1H), 7.33 (d, J = 5.2 Hz, 1H), 6.83 (s, 1H), 5.71 (s, 1H), 5.04-5.01 (m, 1H), 4.78 (s, 2H), 4.64-4.62 (m, 1H), 4.49 (d, J = 2.8 Hz, 1H), 4.32-4.28 (m, 1H), 4.14 (d, J = 4.4 Hz, 2H), 4.09-4.08 (m, 4H), 3.87-3.83 (m, 1H), 3.70 (s, 3H), 2.56 (d, J = 2.8 Hz, 2H), 2.50 (s, 2H), 1.26 (s, 6H).
Example 169a 4-Chloro{6-oxothiaazatricyclo[7.4.0.02,7]trideca- 25 1(9),2(7)-dienyl}pyridinecarbaldehyde 169a 221 O N N N NH O 101l O O S N Cl a)3, S N N O N tricyclohexylphosphine O N Cs2CO3,dioxane, H2O, 169a 110oC, 4 h 169b A 100-mL single-neck round-bottomed flask ed with a reflux condenser was charged with 2-bromochloronicotinaldehyde 103a (1276 mg, 5.80 mmol), 8-thia azatricyclo[7.4.0.02,7}]trideca-1(9),2(7)-dienone 105e (600 mg, 2.90 mmol), CuI (551 mg, 5 2.90 mmol), K2CO3 (800 mg, 5.80 mmol), 4,7-dimethoxy-1,10-phenanthroline (696 mg, 2.90 mmol), and dioxane (20 mL). After bubbling nitrogen through the resulting solution for 10 min, the e was stirred at 95oC for 16 h. It was then cooled to room temperature and ed. To the residue was added water (20 mL). The aqueous layer was separated and ted with ethyl acetate (3 × 20 mL). The combined organic layer was washed with brine 10 (50 mL) and dried over sodium sulfate. The drying agent was d by filtration and the filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 10:1 petroleum ether/ethyl acetate to afford 169a (171 mg, 17%). LCMS-ESI: [M+H]+ 347 Example 169b 4-[1-Methyl({5-[4-(oxetanyl)piperazinyl]pyridin 15 yl}amino)oxo-1,6-dihydropyridinyl]{6-oxothiaazatricyclo[7.4.0.02,7]trideca- 1(9),2(7)-dienyl}pyridinecarbaldehyde 169b A 50-mL single-neck round-bottomed flask equipped with a reflux condenser was charged with 169a (150 mg, 0.43 mmol), 3-(5-(2-ethyl(oxetanyl)piperazin yl)pyridineylamino)methyl(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin- 20 one 101l (200 mg, 0.43 mmol), Pd2(dba)3 (37 mg, 0.040 mmol), tricyclohexylphospine (120 mg, 0.43 mmol), Cs2CO3 (281 mg, 0.86 mmol), dioxane (10 mL), and water (0.1 mL).
After three cycles of vacuum/argon flush, the mixture was heated at 110oC for 4 h. It was then filtered and the filtrate was evaporated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 1:3 petroleum/ethyl acetate to afford 169b 25 as a yellow solid (45 mg, 16%). LCMS-ESI: [M+H]+ 652 222 Example 169 2-{3'-Hydroxymethylmethyl[5-(4-oxetanyl-piperazinyl)- pyridinylamino]oxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}-3,4,5,6,7,8-hexahydro-2H- benzo[4,5]thieno[2,3-c]pyridinone 169 A mixture of 169b (45 mg, 0.070 mmol), NaBH4 (8 mg, 0.21) and methanol (5 mL) 5 was stirred at room temperature for 1 h. The reaction mixture was quenched with water (5 mL) and concentrated under reduced pressure. The residue was extracted with dichloromethane (2 X 10 mL). The combined dichloromethane t was trated under reduced pressure. The residue was purified with e-phase PLC to afford 169 (14 mg, 30%). LCMS-ESI: [M+H]+ 654. 1H NMR (500 MHz, DMSO-d6) δ 8.62 (s, 1H), 10 8.50 (d, J = 2.0 Hz, 1H), 8.43 (s, 1H), 7.86 (d, J = 4.5 Hz, 1H), 7.45 (d, J = 2.5 Hz, 1H), 7.39- 7.34 (m, 2H), 7.25-7.22 (m, 1H), 4.95-4.93 (m, 1H), 4.57-4.55 (m, 2H), 4.47-4.41 (m, 4H), 4.19-4.17 (m, 1H), 3.82-3.80 (m, 1H), 3.60 (s, 3H), 3.45-3.43 (m, 1H), 3.32-3.30 (m, 1H), 3.09-3.07 (m, 4H), 3.01-2.90 (m, 1H), 2.89-2.88 (m, 1H), 2.80-2.79 (m, 2H), 2.51-2.50 (m, 1H), 2.40-2.38 (m, 4H), 1.83-1.80 (m, 4H). 15 Example 170b 4-Chloro[(1R,11S)oxo-3,6- diazatetracyclo[9.2.1.02,10.03,8]tetradeca-2(10),8-dienyl]pyridinecarbaldehyde 170b A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with 1,4-dioxane (30 mL), (1S,11R)-3,6- diazatetracyclo[9.2.1.02,10.03,8]tetradeca-2(10),8-dienone 170a (400 mg, 2.0 mmol), 2- 20 bromochloronicotinaldehyde 103a (1.30 g, 6.0 mmol), and potassium acetate (390 mg, 4.0 mmol). After bubbling nitrogen h the resulting mixture for 30 minutes, Xantphos (110 mg, 0.20 mmol) and tris(dibenzylideneacetone)dipalladium(0) (180 mg, 0.20 mmol) were added, and the reaction mixture was heated at 80 oC for 10 h. After this time the reaction was cooled to room temperature and filtered. The filtrate was ioned between ethyl acetate 25 (50 mL) and water (30 mL). The aqueous layer was separated and extracted with ethyl e (3 X 30 mL). The combined organic layer was washed with brine (20 mL) and dried over sodium sulfate. The drying agent was d by filtration and the filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 2:1 petroleum ether/ethyl e to afford 170b (405 mg, 59%) as a yellow solid. 30 MS-ESI: [M+H]+ 342.2 Example 170c 4-[1-Methyl({5-methyl-4H,5H,6H,7H-pyrazolo[1,5- a]pyrazinyl}amino)oxo-1,6-dihydropyridinyl][(1R,11S)oxo-3,6- diazatetracyclo[9.2.1.02,10.03,8]tetradeca-2(10),8-dienyl]pyridinecarbaldehyde 170c 223 A 100-mL round-bottomed flask equipped with a reflux condenser was charged with 170b (150 mg, 0.44 mmol), 1-methyl(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin- 2-ylamino)(4,4,5,5-tetra-methyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 135a (169 mg, 5 0.44 mmol), sodium acetate (72 mg, 0.88 mmol), K3PO4 (234 mg, 0.88 mmol), Pd (dppf) Cl2 (36 mg, 0.044 mmol), acetonitrile (20 mL), and water (1 mL). After bubbling nitrogen through the reaction mixture for 30 minutes, it was heated at 100oC for 3 hours. The reaction mixture was evaporated under d pressure and the residue was ed by silica-gel column chromatography eluting with 25:1 dichloromethane/methanol to afford 170c (146 mg, 10 52%) as a brown solid. MS-ESI: [M+H]+ 565.3 Example 170 2-(3-(hydroxymethyl)(1-methyl(5-methyl-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 7,8,9-hexahydro-6,9-methanopyrazino[1,2-a]indol-1(2H)-one 170 A solution of 170c (122 mg, 0.22 mmol) in methanol (20 mL) was added NaBH4 (24 15 mg, 0.64 mmol). The mixture was stirred at 20oC for 2 h. The reaction was evaporated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford the title compound (98 mg, 75%) as a white solid. MS-ESI: [M+H]+ 567.3. 1H NMR (500 MHz, DMSO-d6) δ 8.48 (d, J = 5.0 Hz, 1H), 8.19 (s, 1H), 8.05 (d, J = 2.5 Hz, 1H), 7.40 (s, 1H), 7.32 (d, J = 5.0 Hz, 1H), 6.53 (s, 1H), 5.89 (s, 1H), 4.98 (t, J = 5.0 Hz, 1H), 4.48-4.30 (m, 20 3H), 4.27-4.22 (m, 2H), 3.92-3.91 (m, 2H), 3.86-3.84 (m, 1H), 3.59 (s, 3H), 3.49-3.47 (m, 3H), 3.30-3.28 (m, 1H), 2.79-2.77 (m, 2H), 2.35 (s, 3H), .76 (m, 3H), 1.61-1.59 (m, 1H), 1.09-0.88 (m, 2H).
Example 171a 4-[1-Methyl({5-[(2S)methyl(oxetanyl)piperazin yl]pyridinyl}amino)oxo-1,6-dihydropyridinyl][(1S,11R)oxo-3,6- 25 diazatetracyclo[9.2.1.02,10.03,8]tetradeca-2(10),8-dienyl]pyridinecarbaldehyde 171a 224 A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was d with acetonitrile (30 mL), 4-chloro[(1S,11R)oxo-3,6- diazatetracyclo[9.2.1.02,10.03,8]tetradeca-2(10),8-dienyl]pyridinecarbaldehyde 167g (170 5 mg, 0.50 mmol), (S)methyl(5-(2-methyl(oxetanyl)piperazinyl)pyridin o)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 130f (336 mg, 0.70 mmol), water (3 mL ), and potassium acetate (147 mg, 1.5 mmol). After bubbling argon through the suspension for 30 minutes, 1,1’- bis(diphenylphosphino)ferrocenedichloropalladium(II) (408 mg, 0.05 mmol) was added. The 10 system was subjected to three cycles of vacuum/argon flush and heated at 80°C for 3 h. It was then cooled to room temperature and filtered. The solid was washed with dichloromethane (2 X 100 ml). The combined filtrate was trated under reduced pressure and the residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (50:1 to 30:1) to afford 171a (95 mg, 29 %) as a light yellow solid. MS-ESI: [M+H]+ 661.3 15 Example 171 (S)(3-(hydroxymethyl)(1-methyl(5-(2-methyl(oxetan yl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 3,4,6,7,8,9-hexahydro-6,9-methanopyrazino[1,2-a]indol-1(2H)-one 171 To a solution of 171a (90 mg, 0.136 mmol) in methanol (10 mL) was added NaBH4 (26 mg, 0.7 mmol) at room temperature. After the reaction was stirred for 1h, LCMS 20 ted the reaction was complete. It was ed with water (30 mL) and concentrated under reduced pressure. The residue was extracted with dichloromethane (3 X 30 mL). The combined organic layer was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced re. The residue solid was purified by e-phase prep- HPLC to afford 171 (35 mg, 31.5 %) as light yellow solid. MS-ESI: [M+H]+ 663.3. 1H NMR 25 (500 MHz, DMSO-d6) δ 8.62 (s, 1H), 8.48 (d, J = 5.0 Hz, 1H), 8.44 (s, 1H), 7.83 (d, J = 2.5 Hz, 1H), 7.47 (s, 1H), 7.38 (dd, J = 2.5, 9.0 Hz 1H), 7.34 (d, J = 5.0 Hz, 1H), 7.24 (d, J = 9.5 Hz, 1H), 6.51 (s, 1H), 4.97 (t, J = 4.5 Hz, 1H), 4.58-4.54 (m, 2H), 4.50-4.37 (m, 4H), 4.30- 4.24 (m, 2H), 3.86-3.84 (m, 1H), 3.69-3.67 (m, 1H), 3.60 (s, 3H), 3.47 (s, 1H), 3.42-3.37 (m, 225 1H), 3.30 (s, 2H), 3.10-3.07 (m, 1H), 2.95-2.92 (m, 1H), 2.36-2.29 (m, 3H), 2.21-2.16 (m, 1H), 1.88-1.754 (m, 3H), 1.60-1.58 (m, 1H), 1.08-1.05 (m, 1H), 0.98-0.96 (m, 1H), 0.93 (d, J = 6.0 Hz, 3H).
Example 172a 3-(1,2,4-Triazinylamino)bromomethylpyridin-2(1H)- 5 one 172a A 500-mL single-neck bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with oxane (100 mL), 1,2,4-triazinamine (1.5 g, 15.6 mmol), 3,5-dibromomethylpyridin-2(1H)-one (4.2 g, 15.6 mmol), Pd2(dba)3 (458 mg, 1.56 10 mmol), XantPhos (1.8 g, 3.12 mmol), and cesium carbonate (10 g, 31.2 mmol). After three cycles of vacuum/argon flush, the mixture was stirred at 90oC for 2.5 h. After this time the reaction was filtered and the filtrate was evaporated in vacuo. The resulting residue was recrystalized from ethyl acetate to afford 172a as a yellow solid (1.76 g, 40%). MS-ESI: [M+H]+ 282. 15 Example 172b (4-(5-(1,2,4-Triazinylamino)methyloxo-1,6- dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridin- ethyl Acetate 172b A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 172a (200 mg, 0.71 mmol), 3-(acetoxymethyl)(1-oxo- 20 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinylboronic acid 113i (272 mg, 0.71 mmol), Pd(dppf)Cl2 (58 mg, 0.071 mmol), sodium acetate (193 mg, 1.42 mmol), K3PO4 (321 mg, 1.42 mmol), water (0.5 mL) and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the mixture was stirred at 100oC for 3 h. After this time the reaction was filtered and the filtrate was evaporated in vacuo. The resulting e was recrystallized 25 from ethyl acetate to afford 172b as yellow solid (380 mg, 99 %). MS-ESI: [M+H]+ 541.2 Example 172 2-(4-(5-(1,2,4-triazinylamino)methyloxo-1,6-dihydropyridin- 3-yl)(hydroxymethyl)pyridinyl)-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 172 226 A mixture of 172b (350 mg, 0.65 mmol) and lithium hydroxide (273 mg, 6.5 mmol) in i-propanol/THF (1:1, 5 mL) and water (0.5 mL) was stirred at 36oC for 0.5 h. It was then cooled to room temperature and filtered. The te was concentrated under d pressure and the resulting residue was washed by reverse-phase prep-HPLC to afford 172 (90 mg, 5 28%). MS-ESI: [M+H]+ 499.2. 1H NMR (500 MHz, CDCl3) δ 8.77 (d, J = 2.5 Hz, 1H), 8.72 (d, J = 2.0 Hz, 1H), 8.67 (s, 1H), 8.50 (d, J = 5.0 Hz, 1H), 8.34 (d, J = 2.0 Hz, 1H), 8.05 (d, J = 2.5 Hz, 1H), 7.35 (d, J = 5.0 Hz, 1H), 6.89 (s, 1H), 5.10 (t, J = 6.5 Hz, 1H), 4.65-4.51 (m, 2H), 4.31-4.27 (m, 1H), 4.16-4.08 (m, 2H), 3.90-3.87 (m, 1H), 3.75 (s, 3H), 2.62-2.56 (m, 4H), 1.92-1.87 (m, 2H), 1.79-1.78 (m, 2H). 10 Example 173a o(2,6-dimethylpyrimidinylamino) methylpyridin-2(1H)-one 173a A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was d with 1,4-dioxane (150 mL), 2,6-dimethylpyrimidinamine 15 (2.5 g, 20.3 mmol), 3,5-dibromomethylpyridin-2(1H)-one (5.4 g, 20.3 mmol), Pd2(dba)3 (1.86mg, 2.03 mmol), XantPhos (2.3 g, 4.06 mmol), and cesium carbonate (13.2 g, 40.6 mmol). After three cycles of vacuum/argon flush, the e was heated at 90oC for 2.5 h.
After this time the reaction was filtered and the filtrate was evaporated in vacuo. The resulting residue was recrystallized from ethyl acetate to afford 173a as a yellow solid (4.4 g, 20 40%). MS-ESI: [M+H]+ 309.0.
Example 173b 3-(2,6-Dimethylpyrimidinylamino)methyl(4,4,5,5- tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 173b A 250-mL single-neck round-bottomed flask equipped with a magnetic r and a reflux condenser was charged with 173a (1.5 g, 4.9 mmol), Pin2B2 (6.2 g, 24.5 mmol), 25 Pd2(dba)3 (449 mg, 0.49 mmol), X-phos (467 mg, 0.98 mmol), potassium acetate (1.4 g, 14.7 mmol), and dioxane (60 mL). After three cycles of vacuum/argon flush, the mixture was heated at 65ºC for 16 h. The reaction was filtered and the filtrate was evaporated in vacuo.
The resulting residue was recrystallized from ethyl acetate to afford 173b as a light gray solid (1.2 g, 72%). MS-ESI: [M+H]+ 357.2. 227 Example 173c 2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{5-[(2,6-dimethylpyrimidinyl)amino]methyloxo-1,6- opyridinyl}pyridinecarbaldehyde 173c A 100-mL round-bottomed flask equipped with a magnetic stirrer and a reflux 5 condenser was d with 173b (250 mg, 0.70 mmol), 4-chloro{4,4-dimethyloxo- 1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 108a (240 mg, 0.70 mmol), Pd(dppf)Cl2 (57 mg, 0.071 mmol), sodium acetate (19 mg, 1.4 mmol), K3PO4 (316 mg, 1.4 mmol), water (0.5 mL), and acetonitrile (15 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 3 h. After this time the reaction was 10 filtered and the filtrate was evaporated in vacuo. The resulting residue was recrystallized from ethyl e to afford 173c as a brown solid (300 mg, 80%). MS-ESI: [M+H]+ 538.3.
Example 173 2-[5-(2,6-Dimethyl-pyrimidinylamino)-3'-hydroxymethylmethyl- 6-oxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl]-7,7-dimethyl-3,4,7,8-tetrahydro-2H,6H- enta[4,5]pyrrolo[1,2-a]pyrazinone 173 15 At 0 oC, to a solution of 173c (290 mg, 0.54 mmol) in methanol (5 mL) was added sodium borohydride (62 mg, 1.62 mmol). The reaction mixture was stirred at room temperature for 20 minutes and quenched with water (1 mL). It was then trated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 173 as white solid (50 mg, 17 %). MS-ESI: [M+H]+ 540.3. 1H NMR (500 MHz, CDCl3) δ 8.90 (d, J 20 = 2.5 Hz, 1H), 8.51 (d, J = 5.0 Hz, 1H), 8.01 (d, J = 2.0 Hz, 1H), 8.00 (s, 1H), 7.35 (d, J = 5.5 Hz, 1H), 6.85 (s, 1H), 6.45 (s, 1H), 5.16-5.13 (m, 1H), 4.67-4.52 (m, 2H), 4.33-4.29 (m, 1H), 4.16 (d, J = 5.0 Hz, 2H), 3.90-3.86 (m, 1H), 3.72 (s, 3H), 2.58-2.56 (m, overlap, 5H), 2.51 (s , 2H), 2.40 (s, 3H), 2.02 (s, 6H).
Example 174a 4-[1-Methyl({5-[(2S)methyl(oxetanyl)piperazin 25 yl]pyridinyl} amino)oxo-1,6-dihydropyridinyl][(1R,11S)oxo-3,6- diazatetracyclo[9.2.1.02,10.03,8]tetradeca-2(10),8-dienyl]pyridinecarbaldehyde 174a 228 A round-bottomed flask was charged with 4-chloro[(1R,11S)oxo-3,6- diazatetracyclo[9.2.1.02,10.03,8]tetradeca-2(10),8-dienyl]pyridinecarbaldehyde 170b (200 mg, 0.59 mmol), 1-methyl({5-[(2S)methyl(oxetanyl)piperazinyl]pyridin- 2-yl}amino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)-1,2-dihydropyridinone 191j 5 (400 mg, 0.88 mmol), PdCl2(dppf) (50 mg, 0.06 mmol), K3PO4 3water ( 300 mg, 1.20 mmol), sodium acetate (100 mg, 1.20 mmol), acetonitrile (15 mL), and water (1.5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100 oC for 3 h. It was then filtered and the te was evaporated under reduced pressure. The residue was purified with silicagel column chromatography eluting with 1:20 ol/dichloromethane to afford 174a as a 10 red solid (170 mg, 44%). MS-ESI: [M+H]+ 661.3 Example 174 (1R,11S)[3-(Hydroxymethyl)[1-methyl({5-[(2S)methyl (oxetanyl)piperazinyl]pyridinyl}amino)oxo-1,6-dihydropyridinyl]pyridin yl]-3,6-diazatetracyclo[9.2.1.02,10.03,8]tetradeca-2(10),8-dienone 174 A mixture of 174a (150 mg, 0.23 mmol), NaBH4 (34 mg, 0.90), and ol (10 15 mL) was stirred at room temperature for 1 h. The mixture was quenched with water (30ml) and concentrated under reduced pressure. The residue was ted with dichloromethane (2 X 10 mL). The combined dichloromethane extract was concentrated under reduced pressure and the residue was purified with reverse-phase prep-HPLC to afford 174 (42 mg, 28%). MSESI : [M+H]+ 663.3. 1H NMR (500 MHz, CDCl3) δ 8.65 (d, J = 3.0 Hz, 1H), 8.48 (d, J = 6.0 20 Hz, 1H), 7.96 (d, J = 2.5 Hz, 1H), .84 (m, 2H), 7.36 (d, J = 6.5 Hz, 1H), 7.32 (dd, J = 3.5, 11.0 Hz, 1H), 6.82-6.80 (m, 2H), 5.16-5.06 (m, 1H), 4.72-4.61 (m, 5H), .05 (m, 1H), 4.32-4.21 (m, 3H), 3.88-3.85 (m, 1H), 3.71 (s, 3H), 3.54-3.50 (m, 2H), 3.38-3.37 (m, 2H), 3.08-3.06 (m, 2H), 2.57-2.54 (m, 1H), 2.48-2.45 (m, 2H), 2.21-2.17 (m, 1H), 1.93-1.91 (m, 3H), 1.66-1.64 (m, 1H), 1.14-1.08 (m, 2H), 0.98 (d, J = 8.0 Hz, 3H). 25 Example 175a 4-Chloro{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca- 1(9),2(7),3-trienyl}pyridinecarbaldehyde 175a O N N N NH O O N S N N O N 175a 229 A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 4-chloro{6-oxothia-4,5- diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridinecarbaldehyde 124a (150 mg, 0.43 mmol), 3-(5-(2-ethyl(oxetanyl)piperazinyl)pyridinylamino)methyl 5 (4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 161f (215 mg, 0.43 mmol), PdCl2(dppf) (33 mg, 0.040 mmol), K3PO4 trihydrate (202 mg, 0.86 mmol), sodium acetate (71 mg, 0.86 mmol), itrile (10 mL), and water (2 mL). After three cycles of /argon flush, the mixture was heated at 100oC for 3 h. It was then filtered and the filtrate was evaporated under d pressure. The residue was purified by silica-gel column 10 chromatography eluting with 1:3 petroleum/ethyl acetate to afford 175a as a yellow solid (108 mg, 37%). MS-ESI: [M+H]+ 679 Example 175 5-((S)Ethyloxetanyl-piperazinyl)-pyridin ylamino]-3'-hydroxymethylmethyloxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}-6,7,8,9- tetrahydro-3H-benzo[4,5]thieno[2,3-d]pyridazinone 175 15 A mixture of 175a (200 mg, 0.16 mmol), NaBH4 (18 mg, 0.48), and methanol (8 mL) was stirred at 25oC for 1 h. Then the reaction mixture was quenched with water (10 mL) and evaporated under reduced pressure. The residue was extracted with dichloromethane (2 X 10 mL). The combined dichloromethane t was concentrated under reduced pressure and the residue was purified with reverse-phase prep-HPLC to afford 175 (55 mg, 50%). MS-ESI: 20 [M+H]+ 681. 1H NMR (500 MHz, DMSO-d6) 8.61 (d, J = 2.0 Hz, 1H), 8.57-8.56 (m, 1H), 8.47 (s, 1H), 8.43 (s, 1H), 7.83 (d, J = 3.0 Hz, 1H), 7.54-7.53 (m, 1H), 7.47 (d, J = 2.0 Hz, 1H), 7.35-7.32 (m, 1H), 7.24-7.22 (m, 1H), 4.85-4.83 (m, 1H), 4.59-4.55 (m, 2H), 4.47-4.44 (m, 1H), 4.40-4.36 (m, 2H), 3.60 (s, 3H), .49 (m, 1H), 3.40-3.38 (m, 1H), 3.17-3.14 (m, 1H), 3.00-2.95 (m, 3H), 2.87-2.85 (m, 2H), 2.66-2.60 (m, 1H), 2.55-2.53 (m, 1H), .15 25 (m, 1H), 2.10-2.06 (m, 1H), 1.89-1.86 (m, 4H), 1.68-1.64 (m, 1H), 1.28-1.25 (m, 2H), 0.79 (t, J = 9.5 Hz, 3H).
Example 176a (S)(1-Methyl(5-(2-methyl(oxetanyl)piperazin yl)pyridinylamino)oxo-1,6-dihydropyridinyl)(1-oxo-6,7,8,9- tetrahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 176a 230 A round-bottomed flask was charged with 4-chloro(1-oxo-6,7,8,9- tetrahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 121a (180 mg, 0.55 mmol), (S) methyl- 3-(5-(2-methyl(oxetanyl)piperazinyl)pyridinylamino)(4,4,5,5- 5 tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 191j (397 mg, 0.82 mmol), PdCl2(dppf) (45 mg, 0.06 mmol), K3PO4 trihydrate ( 286 mg, 1.10 mmol), sodium acetate (90 mg, 1.10 mmol), acetonitrile (15 mL), and water (1.5 mL). After three cycles of /argon flush, the mixture was heated at 100 oC for 3 h. It was then filtered and the filtrate was ated under reduced pressure. The residue was purified by silica-gel column 10 chromatography eluting with 1:20 methanol/dichloromethane to afford 176a as a red solid (228 mg, 64%). MS-ESI: [M+H]+ 647.3 Example 176 (3-(hydroxymethyl)(1-methyl(5-(2-methyl(oxetan yl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)-6,7,8,9- tetrahydropyrazino[1,2-a]indol-1(2H)-one 176 15 A mixture of 176a (200 mg, 0.31 mmol), NaBH4 (47 mg, 1.20), and methanol (10 mL) was stirred at room temperature for 1 h. The reaction mixture was then quenched with water (10mL) and concentrated under reduced pressure. The residue was extracted with romethane (2 x 10 mL). The combined dichloromethane extract was concentrated under reduced pressure and the residue was purified with reverse-phase prep-HPLC to afford 20 176 (42 mg, 28%). MS-ESI: [M+H]+ 649.3. 1H NMR (500 MHz, DMSO-d6) δ 8.66 (d, J = 2.5 Hz, 1H), 8.56 (d, J = 5.5 Hz, 1H), 8.47 (s, 1H), 7.84 (d, J = 2.5 Hz, 1H), 7.50-7.48 (m, 2H), 7.39-7.36 (m, 1H), 7.26-7.24 (m, 2H), 6.83-6.80 (m, 2H), 4.57-4.54 (m, 2H), 4.48-4.40 (m, 3H), 4.35-4.33 (m, 1H), 3.69-3.67 (m, 1H), 3.60 (s, 3H), 3.41-3.38 (m, 2H), 3.11-3.08 (m, 1H), 2.97-2.93 (m, 1H), 2.76-2.74 (m, 2H), 2.62-2.60 (m, 2H), 2.52-2.51 (m, 1H), 2.35-2.32 25 (m, 2H), .17 (m, 1H), 1.90-1.87 (m, 2H), .75 (m, 2H), 0.94 (d, J = 6.5 Hz, 3H).
Example 177a (4-(5-(5-Ethyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin ylamino)methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl Acetate 177a 231 N N N NH AcO O N N N O N 177a A 50-mL -neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 5-bromo(5-ethyl-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinylamino)methylpyridin-2(1H)-one 208c (300 mg, 0.85 mmol), 3- 5 (acetoxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridin nic acid 113i (1.21 g, 3.16 mmol), PdCl2(dppf) (35 mg, 0.043 mmol), K3PO4 (361 mg, 1.70 mmol), sodium acetate (140 mg, 1.70 mmol), acetonitrile (10 mL), and water (0.5 mL).
The system was evacuated and refilled with N2. The on mixture was stirred at 100ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under 10 reduced pressure and the resulting residue was purified by silica-gel column tography eluting with 30:1 dichloromethane/methanol to afford 177a (365 mg, 60%) as a brown oil.
MS-ESI: [M+H]+ 611 Example 177 2-(4-(5-(5-ethyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinylamino)- 1-methyloxo-1,6-dihydropyridinyl)(hydroxymethyl)pyridinyl)-3,4,6,7,8,9- 15 hexahydropyrazino[1,2-a]indol-1(2H)-one 177 To a solution of 177a (300 mg, 0.49 mmol) in propanol (4 mL), tetrahydrofuran (4 mL), and water (1 mL) was added lithium hydroxide (35 mg, 1.47 mmol). The mixture was stirred at room temperature for 0.5 h. It was evaporated and the e was purified by reverse-phase prep-HPLC to afford 177 (79 mg, 28 %) as a white solid. MS-ESI: [M+H]+ 20 569. 1H NMR (500 MHz, CDCl3) δ 8.46 (d, J = 5.5 Hz, 1H), 7.92 (d, J = 2.0 Hz, 1H), 7.70 (d, J = 2.0 Hz, 1H), 7.39 (s, 1H), 7.33 (d, J = 5.5 Hz, 1H), 6.88 (s, 1H), 5.69 (s, 1H), 4.99- 4.97 (m, 1H), 4.61-4.60 (m, 1H), 4.48-4.46 (m, 1H), 4.33-4.31 (m, 1H), 4.14-4.06 (m, 4H), 3.87-3.85 (m, 1H), 3.69 (s, 3H), 3.62 (d, J = 5.5 Hz, 2H), 2.91 (d, J = 5.0 Hz, 2H), 2.63-2.55 (m, 6H), 1.91-1.87 (m, 2H), .78 (m, 2H), 1.17 (t, J = 7.5 Hz, 3H). 25 Example 178a 5-Bromomethyl(pyridinylamino)pyridin-2(1H)-one 178a 232 A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 1,4-dioxane (60 mL), onitropyridine (8.0 g, 31.8 mmol), pyridinamine (1.0 g, 10.6 mmol), and cesium carbonate (7.0 g, 21.2 mmol). 5 After bubbling nitrogen through the resulting mixture for 30 minutes, XantPhos (616 mg, 1.0 mmol) and ibenzylideneacetone)dipalladium(0) (973 mg, 1.0 mmol) were added. The reaction e was subjected to three cycles of vacuum/argon flush and heated at 100oC for 12 h. After this time the reaction was cooled to room temperature and filtered. The filtrate was partitioned between ethyl acetate (100 mL) and water (100 mL). The aqueous layer was 10 ted and extracted with ethyl acetate (3 X 150 mL). The ed organic layer was washed with brine (50 mL) and dried over sodium sulfate. The drying agent was removed by filtration and the filtrate was concentrated under reduced pressure. The residue was purified on silica-gel column eluting with 30:1 dichloromethane/methanol to afford 178a (1.5 g, 51%) as yellow solid. MS: [M+H]+ 280 15 Example 178b 1-Methyl(pyridinylamino)(4,4,5,5-tetramethyl-1,3,2- dioxaborolanyl)pyridin-2(1H)-one 178b A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 178a (1.06 g, 3.8 mmol), Pin2B2 (4.8 g, 19.0 mmol), Pd2(dba)3 (348 mg, 0.38 mmol), X-Phos (350 mg, 0.76 mmol), potassium acetate (1.12 g, 20 11.40 mmol), and dioxane (30 mL). After three cycles of /argon flush, the mixture was heated at 60ºC for 6 h. It was then cooled to room temperature and filtered. The te was concentrated under reduced pressure and the resulting e was ed by silica-gel column chromatography eluting with 3:1 petroleum ether/ethyl acetate to afford 178b as yellow solid (1.2 g, 96%). MS-ESI: [M+H]+ 328.2 25 Example 178c 4-{1-Methyloxo[(pyridinyl)amino]pyridinyl}{6- oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridinecarbaldehyde 178c A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 178b (131 mg, 0.40 mmol), 4-chloro{6-oxothia-4,5- 233 diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridinecarbaldehyde 124a (138 mg, 0.40 mmol), Pd(dppf)Cl2 (33 mg, 0.040 mmol), K3PO4 (170 mg, 0.80 mmol), sodium acetate (66 mg, 0.80 mmol), water (6 drops), and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at reflux for 2 h. It was then cooled to room 5 temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography g with 30:1 dichloromethane/methanol to afford 178c as a yellow solid (180 mg, 88%). MS-ESI: [M+H]+ 511.2 Example 178 3-[3'-Hydroxymethylmethyloxo(pyridinylamino)-1,6- 10 dihydro-[3,4']bipyridinyl-2'-yl]-6,7,8,9-tetrahydro-3H-benzo[4,5]thieno[2,3-d]pyridazin one 178 To a solution of 178c (179 mg, 0.35 mmol) in methanol (6 mL) was added sodium borohydride (39 mg, 1.05 mmol) at 0 oC. The reaction mixture was stirred for 30 s and quenched with water (1.0 mL). It was then concentrated under reduced re and the 15 residue was ed by e-phase prep-HPLC to afford 178 (100 mg, 56 %). MS-ESI: [M+H]+ 513.3. 1H NMR (500 MHz, CDCl3) δ 8.81 (s, 1H), 8.68 (d, J = 5.0 Hz, 1H), 8.32 (s, 1H), 8.27 (d, J = 4.0 Hz, 1H), 7.94 (bs, 1H), 7.76 (d, J = 1.5 Hz, 1H), 7.58-7.56 (m, 2H), 6.85-6.80 (m, 2H), 4.47-4.45 (m, 2H), 4.38-4.36 (m, 1H), 3.74 (s, 3H), 3.01-2.99 (m, 2H), 2.89-2.87 (m, 2H), 2.02-1.99 (m, 4H). 20 Example 179a 4-(1-Methyl(2-methylpyrimidinylamino)oxo-1,6- dihydropyridinyl)(4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dien- 10-yl)nicotinaldehyde 179a N N NH O O N N N O N 179a A 50-mL round-bottomed flask equipped with a reflux ser was charged with 1- 25 methyl(2-methylpyrimidinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolan yl)pyridin-2(1H)-one 213b (510 mg, 1.5 mmol), 4-chloro{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 108a (343 mg, 1.0 mmol), K3PO4 (424 mg, 2.0 mmol), sodium acetate (272 mg, 2.0 mmol), 1,1’- bis(diphenylphosphino)ferrocenedichloropalladium(II) (40 mg, 0.044 mmol), acetonitrile (20 234 mL), and water (0.5 mL). After bubbling nitrogen through the mixture for 30 minutes, it was was heated at 100°C under N2 protection for 2 h. Analysis of the reaction mixture by LCMS showed complete conversion to the desired product. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue 5 was diluted with dichloromethane (30 mL) and water (30 mL). The aqueous layer was separated and extracted with dichloromethane (3 × 20 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under d pressure. The dark e was purified by silica-gel column chromatography eluting with dichloromethane/methanol (80/1 to 30/1) to afford 179a (300 mg, 57%) as a yellow solid. MS-ESI: [M+H]+ 524 10 e 179 Hydroxymethylmethyl(2-methyl-pyrimidinylamino) oxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl]-7,7-dimethyl-3,4,7,8-tetrahydro-2H,6H- cyclopenta[4,5]pyrrolo[1,2-a]pyrazinone 179 To a solution of 179a (262 mg, 0.50 mmol) in methanol/dichloromethane(10/10 mL) was added NaBH4 (57 mg, 1.5 mmol) at room temperature. After the reaction was stirred for 15 1h, LCMS indicated the reaction was te. Then the e was concentrated under reduced pressure. The residue was diluted with water (5 mL) and romethane (20 mL.
The water phase was ted and extracted with dichloromethane (3 X 10 mL). The combined c layer was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue solid was purified by reverse-phase prep- 20 HPLC to afford 179 (180 mg, 69%) as white solid. MS-ESI: [M+H]+ 526. 1H NMR (500 MHz, CDCl3) δ 8.93 (d, J = 2.0 Hz, 1H), 8.53 (d, J = 5.5 Hz, 1H), 8.28 (d, J = 5.5 Hz, 1H), 8.07 (s, 1H), 8.03 (d, J = 2.0 Hz, 1H), 7.36 (s, 1H), 6.86 (s, 1H), 6.60 (d, J = 6.0 Hz, 1H), 5.18-5.16 (m, 1H), 4.70-4.67 (m, 1H), 4.55-4.53 (m, 1H), 4.33-4.31 (m, 1H), 4.18-4.16 (m, 2H), 3.91-3.90 (m, 1H), 3.74 (s, 3H), 2.60 (s, 3H), 2.58 (d, J = 5.5 Hz, 2H), 2.52 (s, 2H), 1.28 25 (s, 6H).
Example 180a 5-Bromomethyl(6-methylpyrimidinylamino)pyridin- 2(1H)-one 180a N N N N NH N NH O N NH 108a O O N O O N B Pd2(dba)3, N N Pd(dppf)Cl2, N X-phos, O K3PO4, H2O Br AcOK,Pin2B2, O N NaOAc, MeCN, dioxane, reflux, 3 h 65°C 180a 180b 180c 235 A 100-mL single-neck round-bottomed flask equipped with a magnetic r and a reflux condenser was charged with 6-methylpyrimidinamine (800 mg, 2.6 mmol), 3,5- dibromomethylpyridin-2(1H)-one (694 mg, 2.6 mmol), tris(dibenzylideneacetone)dipalladium(0) (238 mg, 0.26 mmol), XantPhos (300 mg, 0.52 5 mmol), Cs2CO3 (1.7 g, 5.2 mmol), and 1,4-dioxane (30 mL). After three cycles of vacuum/argon flush, the mixture was heated at 90oC for 2.5 h. The mixture was cooled to room temperature and filtered. The filtrate was concentrated under d pressure and the resulting residue was purified by silica-gel column chromatography eluting with 50:1 dichloromethane/methanol to afford 180a as a yellow solid (800 mg, 36%). MS-ESI: [M+H]+ 10 295.1 Example 180b 1-Methyl(6-methylpyrimidinylamino)(4,4,5,5- tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 180b A 100-mL single-neck bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 180a (1.0 g, 3.4 mmol), Pin2B2 (4.3 g, 17 mmol), 15 Pd2(dba)3 (312 mg, 0.34 mmol), X-phos (324 mg, 0.68 mmol), potassium acetate (666 mg, 6.8 mmol), and dioxane (40 mL). After three cycles of vacuum/argon flush, the mixture was heated at 65ºC for 14 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the ing residue was washed with 3:1 petroleum ether/ethyl acetate (80 mL) to afford 180b as a yellow solid (600 mg, 50%). MS-ESI: 20 [M+H]+ 343.2.
Example 180c -Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{1-methyl[(6-methylpyrimidinyl)amino]oxo-1,6- dihydropyridinyl}pyridinecarbaldehyde 180c A 50-mL single-neck round-bottomed flask equipped with a magnetic r and a 25 reflux condenser was charged with 180b (239 mg, 0.70 mmol), ro{4,4-dimethyl oxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 108a ( 239mg 0.70 mmol), Pd(dppf)Cl2 (57 mg , 0.070 mmol), sodium e (115 mg, 1.4 mmol), K3PO4 (320 mg, 1.4 mmol), water (5 mL), and acetonitrile (15 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 3 h. After this time the 30 reaction was filtered and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica-gel column chromatography eluting with 40:1 dichloromethane/methanol to afford 180c as a yellow solid (170 mg, 47%). MS-ESI: [M+H]+ 524.2. 236 Example 180 2-[3'-Hydroxymethylmethyl(6-methyl-pyrimidinylamino) oxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl]-7,7-dimethyl-3,4,7,8-tetrahydro-2H,6H- cyclopenta[4,5]pyrrolo[1,2-a]pyrazinone 180 At 0 oC, to a solution of 180c with water (1 mL). It was then concentrated under 5 reduced re and the residue was purified by reverse-phase PLC to afford 180 (47 mg, 32 %). MS-ESI: [M+H]+ 526.2. 1H NMR (500 MHz, CDCl3) δ 8.82 (d, J = 2.5 Hz, 1H), 8.68 (s, 1H), 8.51 (d, J = 2.5 Hz, 1H), 8.02-8.00 (m, 2H), 7.35 (d, J = 5.0 Hz, 1H), 6.84 (s, 1H), 6.62 (s, 1H), 5.13 (t, J = 6.5 Hz, 1H), 4.67-4.52 (m, 2H), 4.29-4.15 (m, 3H), 3.88-3.86 (m, 1H), 3.72 (s, 3H), 2.57 (d, J = 5.5 Hz, 2H), 2.51 (s, 2H), 2.43 (s, 3H), 1.28 (s, 6H). 10 Example 181a 4-[1-Methyl({5-methyl-4H,5H,6H,7H-pyrazolo[1,5- a]pyrazinyl}amino)oxo-1,6-dihydropyridinyl]{6-oxothia-4,5- diazatricyclo[7.4.0.02,7]trideca-1(9), 2(7),3-trienyl}pyridinecarbaldehyde 181a N N N NH O O N S N N O N 181a A 100-mL single-neck round-bottomed flask equipped with a reflux condenser was 15 charged with ro{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trien yl}pyridinecarbaldehyde 124a (210 mg, 0.60 mmol), 1-methyl(5-methyl-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)(4,4,5,5-tetra-methyl-1,3,2-dioxaborolan yl)pyridin-2(1H)-one 135a (346 mg, 0.90 mmol), Pd(dppf)Cl2 (30 mg, 0.030 mmol), K3PO4 (270 mg, 1.2 mmol), sodium acetate trihydrate (180 mg, 1.2 mmol), water (6 drops), and 20 acetonitrile (40 mL). The system was evacuated and refilled with N2. The reaction mixture was heated at 100 ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the ing residue was purified by silica-gel column chromatography eluting with 25:1 dichloromethane/methanol to afford 181a (300 mg, 88%) as a yellow brown solid. MS-ESI: [M+H]+ 569.3. 25 Example 181 3-[3'-Hydroxymethylmethyl(5-methyl-4,5,6,7-tetrahydropyrazolo [1,5-a]pyrazinylamino)oxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl]-6,7,8,9- tetrahydro-3H-benzo[4,5]thieno[2,3-d]pyridazinone 181 237 A mixture of 181a (300 mg, 0.50 mmol) and NaBH4 (60 mg, 1.5 mmol) in methanol (20 mL) was stirred at 30oC for 1 h. The mixture was quenched with water and trated under reduced pressure. The e was extracted with ethyl e (3 X 10 mL). The ed ethyl acetate extract was concentrated under reduced pressure and the residue was 5 purified by reverse-phase prep-HPLC to afford 181 (100 mg, 35%). MS-ESI: [M+H]+ 571.2. 1H NMR (500 MHz, CHCl δ 8.64 (d, J = 5.0 Hz, 1H), 8.30 (s, 1H), 8.00 (d, J = 2.0 Hz, 3) 1H), 7.57 (d, J = 2.0 Hz, 1H), 7.53 (d, J = 5.0 Hz, 1H), 7.43 (s, 1H), 5.70 (s, 1H), 4.45-4.43 (m, 2H), 4.32 (bs, 1H), 4.11-4.09 (m, 2H), 3.71 (s, 3H), 3.63 (s, 2H), 2.99-2.97 (m, 2H), 2.93- 2.91 (m, 2H), 2.88-2.86 (m, 2H), 2.50 (s, 3H), 2.00-1.98 (m, 4H). 10 Example 182a 4-{1-Methyloxo[(pyrimidinyl)amino]-1,6- dihydropyridinyl}{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trien idinecarbaldehyde 182a N N NH O O N S N N O N 182a A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 15 reflux condenser was charged with ro{6-oxothia-4,5- diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridinecarbaldehyde 124a (345 mg, 1.0 mmol), 1-methyl(pyrimidinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolan yl)pyridin-2(1H)-one 143a (328 mg, 1.0 mmol), Pd(dppf)Cl2 (82 mg, 0.10 mmol), sodium acetate (162 mg, 2.0 mmol), K3PO4 (424 mg, 2.0 mmol), and acetonitrile/water (20/1 mL). 20 After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 3 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 40:1 dichloromethane/methanol to afford 182a as a yellow solid (156 mg, 30%). MSESI : [M+H]+ 512.1. 25 Example 182 3-[3'-Hydroxymethylmethyloxo(pyrimidinylamino)-1,6- dihydro-[3,4']bipyridinyl-2'-yl]-6,7,8,9-tetrahydro-3H-benzo[4,5]thieno[2,3-d]pyridazin one 182 238 At room temperature, to a on of 182a (140 mg, 0.27 mmol) in methanol (5 mL) was added sodium borohydride (31 mg, 0.82 mmol). The reaction mixture was stirred for 20 minutes and quenched with water (1 mL). It was then concentrated under reduced pressure and the ing residue was purified by silica-gel column tography eluting with 60:1 5 dichloromethane/methanol to afford 182 as a white solid (60 mg, 43%). : [M+H]+ 514.2. 1H NMR (500 MHz, CDCl3) δ 8.89 (d, J = 2.5 Hz, 1H), 8.82 (s, 1H), 8.70 (d, J = 5.5 Hz, 1H), 8.36 (d, J = 6.5 Hz, 1H), 8.31 (s, 1H), 8.20 (s, 1H), 7.89 (d, J = 2.0 Hz, 1H), 7.55 (d, J = 5.0 Hz, 1H), 6.80 (d, J = 9.5 Hz, 1H), 4.44-4.42 (m, 3H), 3.75 (s, 3H), .98 (m, 2H), 2.88-2.87 (m, 2H), 2.03-1.98 (m, 4H). 10 Example 183a 2-(10-Fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl)(1-methyloxo(pyridinylamino)-1,6-dihydropyridinyl)nicotinaldehyde 183a A 100-mL flask equipped with a reflux condensor was charged with 4-chloro(10- 15 fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 134c (297 mg, 0.57 mmol), 1-methyl(pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolan- 2-yl)pyridin-2(1H)-one 178b (186 mg, 0.57 mmol), sodium acetate (90 mg, 1.1 mmol), K3PO4 (234 mg, 1.1 mmol), PdCl2(dppf) (50 mg, 0.057 mmol), acetonitrile (25 mL), and water (1 mL). After bubbling nitrogen through the mixture for 30 minutes, it was heated at 20 100oC under nitrogen atmosphere for 3 hours. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica-gel column tography eluting with 20:1 methylene chloride/methanol to afford 183a (178 mg, 61%) as a brown solid. MS-ESI: [M+H]+ 513.3.
Example 183 10-fluoro(3-(hydroxymethyl)(1-methyloxo(pyridin 25 ylamino)-1,6-dihydropyridinyl)pyridinyl)-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 1(2H)-one 183 A mixture of 183a (160 mg, 0.31 mmol) and NaBH4 (59 mg, 1.55 mmol) in methanol (20 mL) was stirred at 20 oC for 2 h. The reaction was then quenched with water and 239 evaporated under reduced re. The residue was purified by reverse-phase prep-HPLC to afford 183 (42 mg, 26%) as an ite solid. MS-ESI: [M+H]+ 515.3. 1H NMR (500 MHz, DMSO-d6) δ 8.74 (d, J = 2.5 Hz, 1H), 8.65 (s, 1H), 8.50 (d, J = 5.0 Hz, 1H), 8.18-8.17 (m, 1H), 7.61-7.58 (m, 1H), 7.53 (d, J = 2.0 Hz, 1H), 7.36 (d, J = 5.0 Hz, 1H), 7.31 (d, J = 8.5 5 Hz, 1H), 6.81-6.79 (m, 1H), 4.95 (t, J = 5.0 Hz, 1H), 4.49-4.40 (m, 2H), 4.22-4.14 (m, 2H), 4.10-4.05 (m, 1H), .85 (m, 1H), 3.62 (s, 3H), 2.64-2.60 (m, 1H), 2.57-2.53 (m, 1H), 2.43-2.41 (m, 2H), 1.81-1.75 (m, 2H), .67 (m, 2H).
Example 184a 1-Methyl(pyrazinylamino)(4,4,5,5-tetramethyl-1,3,2- dioxaborolanyl)pyridin-2(1H)-one 184a 10 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 5-bromomethyl(pyrazinylamino)pyridin-2(1H)- one 162a (600 mg, 2.1 mmol), Pin2B2 (2540 mg, 10 mmol), Pd2(dba)3 (100 mg, 0.11 mmol), X-phos (100 mg, 0.25 mmol), potassium acetate (600 mg, 6.1 mmol), and dioxane (30 mL). 15 After three cycles of vacuum/argon flush, the e was heated at 65ºC for 15 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting e was washed by petroleum ether to afford 184a as a yellow solid (700 mg, 90%). MS-ESI: [M+H]+ 329.4 Example 184b 2-{4,4-Dimethyloxothiaazatricyclo[6.4.0.02,6]dodeca- 20 1(8),2(6)-dienyl}{1-methyloxo[(pyrazinyl)amino]-1,6-dihydropyridin yl}pyridinecarbaldehyde 184b A 25-mL single-neck round-bottomed flask equipped with a reflux condenser was charged with 4-chloro{4,4-dimethyloxothiaazatricyclo[6.4.0.02,6]dodeca- 1(8),2(6)-dienyl}pyridinecarbaldehyde 109a (100 mg, 0.30 mmol), 184a (170 mg, 25 0.60 mmol), Pd(dppf)Cl2 (12 mg, 0.015 mmol), K3PO4 (130 mg, 0.60 mmol), sodium acetate rate (85 mg, 0.60 mmol), acetonitrile (10 mL), and water (6 drops). The system was evacuated and refilled with N2. The reaction mixture was stirred at 100ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure 240 and the resulting residue was purified by silica-gel column chromatography eluting with 25:1 of dichloromethane/methanol to afford 184b (80 mg, 54%) as a yellow brown solid. : [M+H]+ 527.2.
Example 184 6-[3'-Hydroxymethylmethyloxo(pyrazinylamino)-1,6- 5 dihydro-[3,4']bipyridinyl-2'-yl]-2,2-dimethyl-2,3,5,6-tetrahydro-1H,4Hthiaazacyclopenta enone 184 A mixture of 184b (80 mg, 0.15 mmol) and NaBH4 (18 mg, 0.45 mmol) in methanol (5 mL) was stirred at 30oC for 2 h. The mixture was quenched with water and concentrated under reduced pressure. The residue was extracted with ethyl acetate (3 X 10 mL). The 10 combined ethyl acetate extract was concentrated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 184 (24 mg, 35%) as a white solid. MS-ESI: [M+H]+ 529.3. 1H NMR (500 MHz, CHCl3) δ 8.73 (d, J = 2.0 Hz, 1H), 8.54 (d, J = 5.0 Hz, 1H), 8.29 (s, 1H), 8.14 (d, J = 8.0 Hz, 2H), 8.02 (s, 1H), 7.96 (d, J = 2.5 Hz, 1H), 7.37 (d, J = 5.5 Hz, 1H), 4.86-4.83 (m, 1H), 4.71-4.68 (m, 1H), 4.46-4.41 (m, 1H), 4.32 (t, J = 11.0 Hz, 15 1H), .81 (m, 1H), 3.74 (s, 3H), 2.99-2.94 (m, 2H), 2.81 (s, 2H), 2.61-2.51 (m, 2H), 1.30 (s, 6H). e 185a o(imidazo[1,2-a]pyridinylamino) methylpyrazin-2(1H)-one 185a 20 A 100-mL round-bottomed flask equipped with a reflux condenser was charged with imidazo[1,2-a]pyridinamine (665 mg, 5.0 mmol), Cs2CO3 (3.26 g, 10 mmol), 3,5- dibromomethylpyrazin-2(1H)-one (1.86 g, 7.0 mmol), Xantphos (289 mg, 0.50 mmol), Pd2(dba)3 (458 mg, 0.50 mmol), and 1, 4-dioxane (30 mL). After bubbling nitrogen through the mixture for 30 minutes, it was heated at 100°C under nitrogen here for 16 h. 25 Analysis of the reaction mixture by LCMS showed little starting material remained. The reaction mixture was cooled to room temperature and filtered. The filtrate was diluted with dichloromethane (60 mL) and water (50 mL). The aqueous layer was ted and extracted with dichloromethane (3 × 20 mL). The combined organic layers was dried over Na2SO4, filtered, and trated under reduced pressure. The dark residue was purified by silica-gel 241 column chromatography eluting with dichloromethane/methanol (60/1 to 30/1) to afford 185a (700 mg, 44%) as a light yellow solid. MS-ESI: [M+H]+ 320 Example 185b 6-(Imidazo[1,2-a]pyridinylamino)methyloxo-4,5- dihydropyrazinylboronic Acid 185b 5 A 100-mL round-bottomed flask equipped with a reflux condenser was charged with 185a (638 mg, 1.99 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (Pin2B2, 2.54 mg, 10 mmol), 1,1’-bis(diphenylphosphino)ferrocenedichloropalladium(II) (163 mg, 0.18 mmol), Cs2CO3 (1.3 g, 3.98 mmol), and 1,4-dioxane (20 mL). After bubbling nitrogen h the mixture for 30 minutes, it was heated at 80°C under nitrogen atmosphere for 3 h. 10 The reaction mixture was cooled to room temperature and filtered. The filtrate was diluted with petroleum ether (150 mL) and ethyl acetate (15 mL). The ing suspension was stirred at room temperature for 30 s. The solid was collected by filtration and further purified by silica-gel column tography eluting with dichloromethane/methanol (60/1 to 15/1) to afford 185b (400 mg, 70 %) as an off-white solid. MS-ESI: [M+H]+ 286 15 Example 185c -Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dien yl}(6-{imidazo[1,2-a]pyridinylamino}methyloxopyrazin yl)pyridinecarbaldehyde 185c A 100-mL round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 185b (400 mg, 1.40 mmol), 4-chloro{4,4-dimethyloxo- 20 1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 108a (192 mg, 0.56 mmol), potassium acetate (220 mg, 2.24 mmol), itrile (20 mL), and water (0.5 mL). After bubbling nitrogen through the suspension for 30 minutes, 1,1’- bis(diphenylphosphino)Ferrocenedichloropalladium(II) (49 mg, 0.054 mmol) was added. The system was subjected to three cycles of vacuum/argon flush and heated at 80 °C for 3 h. It 25 was then cooled to room temperature and filtered. The solid was washed with dichloromethane (2 X 20 mL). The combined filtrate was concentrated under reduced pressure and the residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (40:1 to 10:1) to afford 185c (90 mg, 29%) as a light yellow solid.
MS-ESI: [M+H]+ 549 30 e 185 2-{3-Hydroxymethyl[6-(imidazo[1,2-a]pyridinylamino) methyloxo-4,5-dihydro-pyrazinyl]-pyridinyl}-7,7-dimethyl-3,4,7,8-tetrahydro- 2H,6H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazinone 185 To a on of 185c (80 mg, 0.146 mmol) in methanol (5 mL) was added NaBH4 (34 mg, 0.90 mmol) at room temperature. After the on was stirred for 1h, LCMS indicated 242 the reaction was complete. The reaction mixture was quenched with water (3 mL) and trated under reduced pressure. The residue was ted with dichloromethane (3 X 10 mL). The combined organic layer was washed with brine (30 mL), dried over Na2SO4, ed, and concentrated under reduced pressure. The residue was purified by reverse-phase 5 prep-HPLC to afford 185 (49 mg, 61%) as light yellow solid. MS-ESI: [M+H]+ 551. 1H NMR (500 MHz, DMSO-d6) δ 9.67 (s, 1H), 8.57 (d, J = 10.5 Hz, 1H), 8.51 (s, 1H), 8.41 (d, J = 7.0 Hz, 1H), 7.77 (s, 1H), 7.75 (s, 1H), 7.63 (s, 1H), 7.46 (d, J = 6.0 Hz, 1H), 7.42 (s, 1H), 6.59 (s, 1H), 5.04-5.02 (m, 1H), 4.67-4.64 (m, 1H)，4.519-4.481 (m, 1H), 4.31-4.20 (m, 3H), 3.88 (d, J = 7.0 Hz, 1H), 3.58 (s, 3H), 2.63-2.55 (m, 2H), 2.44-2.42 (m, 2H), 1.23 (s, 6H). 10 Example 186a 5-Bromomethyl(pyridinylamino)pyrazin-2(1H)-one 186a A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with pyridinamine (940 mg, 10 mmol), 3,5-dibromo 15 methylpyrazin-2(1H)-one (5.4 g, 20 mmol), i-propanol (50 mL), and di-i-propylethylamine (10 mL). The mixture was heated at reflux for 5 h. After the completion of the reaction, it was cooled to room temperature. The solvent was removed under d pressure. The crude was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 186a (1.4 g, 50%) as a yellow solid. : [M+H]+ 20 281.6. e 186b 4-Methyloxo(pyridinylamino)-4,5-dihydropyrazin ylboronic acid 186b A 250-mL round-bottomed flask equipped with a reflux condenser was charged with 186a (800 mg, 2.86 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (2.18 g, 25 8.57 mmol), Pd (dppf) Cl2 (204 mg, 0.28 mmol), potassium acetate (560 mg, 5.71 mmol ), and e (60 mL). After bubbling nitrogen through the mixture for 30 minutes, it was d at 100oC for 3 h under nitrogen. The mixture was cooled to room temperature and filtered. The filtrate was evaporated under reduce pressure. The residue solid was washed with petroleum ether (2 X 30 mL) to afford 186b (406 mg, 58%) as a brown solid. MS-ESI: 30 [M+H]+ 247.3. 243 Example 186c 2-(10-Fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl)(4-methyloxo(pyridinylamino)-4,5-dihydropyrazinyl)nicotinaldehyde 186c A 50-mL round-bottomed flask equipped with a reflux condenser was charged with 5 186b (127 mg, 0.52 mmol), 4-chloro(10-fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)nicotinaldehyde 134c (180 mg, 0.52 mmol), Na2CO3 (110 mg, 1.04 mmol), PdCl2(dppf) (38 mg, 0.052 mmol), DMF (12 mL), and water (1 mL). After ng nitrogen through the e for 30 minutes, it was heated at 50oC for 8 hours under nitrogen. The reaction was then cooled to room temperature and concentrated under reduce pressure. The 10 residue was purified by silica-gel column tography eluting with 30:1 methylene chloride/methanol to afford 186c (132 mg, 49%) as a brown solid. MS-ESI: [M+H]+ 514.3.
Example 186 10-fluoro(3-(hydroxymethyl)(4-methyloxo(pyridin ylamino)-4,5-dihydropyrazinyl)pyridinyl)-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 1(2H)-one 186 15 To a solution of 186c (118 mg, 0.23 mmol) in methanol (15 mL) was added NaBH4 (27 mg, 0.70 mmol). The mixture was d at 20oC for 2 h. The reaction was quenched with water and evaporated under reduced pressure. The residue was purified by reverse-phase prep-HPLC to afford 186 (33 mg, 28%) as a white solid. MS-ESI: [M+H]+ 516.3. 1H NMR (500 MHz, DMSO-d6) δ 9.78 (s, 1H), 8.54 (d, J = 5.0 Hz, 1H), 8.39-8.37 (m, 2H), 8.06-8.04 20 (m, 2H), 7.76 (s, 1H), 7.59 (d, J = 5.5 Hz, 1H), 4.97 (t, J = 5.0 Hz, 1H), 4.68-4.65 (m, 1H), 4.51-4.47 (m, 1H), .19 (m, 2H), 4.10-4.05 (m, 1H), 3.91-3.88 (m, 1H), 3.58 (s, 3H), 2.66-2.60 (m, 1H), 2.57-2.53 (m, 1H), 2.44-2.42 (m, 2H), .75 (m, 2H), 1.71-1.67 (m, 2H).
Example 187a (4-(5-(5-(2,2-Dimethyl(oxetanyl)piperazinyl)pyridin- 25 2-ylamino) methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl acetate 187a 244 A 50-mL single-neck round-bottomed flask ed with a magnetic stirrer and a reflux condenser was charged with 5-bromo(5-(2,2-dimethyl(oxetanyl)piperazin yl)pyridinylamino)methylpyridin-2(1H)-one 190e (200 mg, 1.0 eq., 0.45 mmol), (2-(1- oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)(4,4,5,5-tetramethyl-1,3,2- 5 dioxaborolanyl)pyridinyl)methyl acetate 113i (345 mg, 2 eq., 0.90 mmol), dppf) (36 mg, 0.1 eq., 0.045 mmol), K3PO4 (191 mg, 2 eq., 0.90 mmol), sodium acetate (74 mg, 2.0 eq., 0.90 mmol), acetonitrile (15 mL), and water (0.1 mL). After three cycles of vacuum/argon flush, the mixture was d at 90oC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under d pressure and the 10 resulting residue was purified by silica-gel column chromatography eluting with 50:1 romethane/ethanol to afford 187a (100 mg, 31%) as yellow solid. MS-ESI: [M+H]+ 707.4.
Example 187 2-(4-(5-(5-(2,2-dimethyl(oxetanyl)piperazinyl)pyridin ylamino)methyloxo-1,6-dihydropyridinyl)(hydroxymethyl)pyridinyl)- 15 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 187 A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with 187a (100 mg, 1 eq., 0.14 mmol), lithium hydroxide (54 mg, 10 eq., 1.4 mmol), i-propanol (3 mL), THF (3 mL) and water (2 mL). The mixture was stirred at 30oC for 1 h. It was then filtered and the filtrate was concentrated under reduced re. The residue was 20 purified by reverse-phase prep-HPLC to afford 187 as a white solid (43 mg, 46%). MS-ESI: [M+H]+ 665.3. 1H NMR (500 MHz, DMSO-d6) δ 8.69 (d, J = 2.0 Hz, 1H), 8.60 (s, 1H), 8.48 (d, J = 5.0 Hz, 1H), 7.93 (d, J = 2.0 Hz, 1H), 7.51(d, J = 1.5 Hz, 1H), 7.42-7.40 (m, 1H), 7.35 (d, J = 5.0 Hz, 1H), 7.24 (d, J = 8.5 Hz, 1H), 6.58 (s, 1H), 4.98 (brs, 1H), 4.54 (t, J = 6.0 Hz, 2H), 4.46-4.38 (m, 4H), 4.25-3.85 (m, 4H), 3.60 (s, 3H), 3.38-3.35 (m, 1H), 3.03-2.54 25 (m, 4H), 2.47 (t, J = 6.0 Hz, 2H), 2.32-2.12 (m, 4H), 1.79-1.67 (m, 4H), 0.97 (s, 6H).
Example 188a (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{1-methyloxo[(pyrazinyl)amino]-1,6-dihydropyridin yl}pyridinyl)methyl Acetate 188a 245 A 50-mL single-neck round-bottomed flask equipped with a reflux condenser was charged with 5-bromomethyl(pyrazinylamino)pyridin-2(1H)-one 162a (210 mg, 0.70 mmol), {3-[(acetoxy)methyl]{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- -dienyl}pyridinyl}boronic acid 199e (560 mg, 1.4 mmol), Pd(dppf)Cl2 (70 mg, 5 0.035 mmol), K3PO4 (320 mg, 1.4 mmol), sodium e trihydrate (210 mg, 1.4 mmol), acetonitrile (10 mL), and water (6 drops). The system was evacuated and refilled with N2.
The reaction mixture was stirred at 100 ºC for 2 h. It was then cooled to room temperature and ed. The filtrate was concentrated under reduced pressure and the resulting e was purified by -gel column chromatography eluting with 25:1 of 10 dichloromethane/methanol to afford 188a (150 mg, 40%) as a yellow brown solid. MS-ESI: [M+H]+ 554.2.
Example 188 2-[3'-Hydroxymethylmethyloxo(pyrazinylamino)-1,6- dihydro-[3,4']bipyridinyl-2'-yl]-7,7-dimethyl-3,4,7,8-tetrahydro-2H,6H- cyclopenta[4,5]pyrrolo[1,2-a]pyrazinone 188 15 A mixture of 188a (150 mg, 0.27 mmol) and lithium hydroxide (103 mg, 2.7 mmol) in i-propanol /THF (5:3, 8 mL) and water (2 mL) was stirred at 30 oC for 1 h. The mixture was evaporated in vacuo and the residue was extracted with ethyl e (2 X 20 mL). The combined ethyl acetate extract was concentrated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 188 (40 mg, 35%) as a white solid. MS-ESI: 20 [M+H]+ 512.3. 1H NMR (500 MHz, CHCl3) δ 8.73 (d, J = 2.0 Hz, 1H), 8.53 (d, J = 5.0 Hz, 1H), 8.29 (s, 1H), 8.15-8.13 (m, overlap, 2H), 8.02-8.00 (m, 2H), 7.38 (d, J = 5.0 Hz, 1H), 6.86 (s, 1H), 5.12 (s, 1H), 4.68-4.51 (m, 2H), 4.33-4.29 (m, 1H), 4.18 (t, J = 5.5 Hz, 2H), 3.91-3.86 (m, 1H), 3.75 (s, 3H), 2.60-2.58 (m, 2H), 2.53 (s, 2H), 1.28 (s, 6H).
Example 189a tert-Butyl 4-(Pyrazinyl)piperazinecarboxylate 189a 25 A 500-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with DMSO (250 mL), tert-butyl piperazinecarboxylate (15.8 g, 85.0 mmol), ropyrazine (9.7 g, 85.0 mmol), and Cs2CO3 (55.3 g, 170 mmol).
The mixture was heated at 60oC for 3 days. After this time the reaction was cooled to room temperature. It was then filtered and the filtrate was evaporated under reduced pressure. The 30 residue was purified by silica-gel column chromatography eluting with 5:1 petroleum ether/ethyl acetate to afford 189a (13.3 g, 60 %) as a yellow solid. MS: [M+H]+ 265.3 Example 189b tert-Butyl 4-(5-Bromopyrazinyl)piperazinecarboxylate 189b 246 A 500-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with acetonitrile (150 mL), 189a (3.0 g, 8.8 mmol), and N- bromosuccinimide (1.56 g, 8.8 mmol). The mixture was d at room ature overnight. It was then concentrated under reduced pressure and the resulting residue was 5 purified by silica-gel column chromatography eluting with 10:1 petroleum ethyl acetate to afford 189b as a yellow solid (2.85 g, 73.4 %). MS: [M+H]+ 343.3. 1H NMR (500 MHz, (CD3)2CO) δ 8.03 (s, 1H), 7.94 (s, 1H), 3.48-3.46 (m, 4H), 3.42-3.40 (m, 4H), 1.33 (s, 9H).
Example 189c tert-Butyl 4-(5-(Diphenylmethyleneamino)pyrazin yl)piperazinecarboxylate 189c 10 A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 189b (3.3 g, 9.6 mmol), diphenylmethanimine (1.74 g, 9.6 mmol), palladium diacetate (440 mg, 0.48 mmol), (R)-(+)-2,2'-bis(diphenylphosphino)-1,1'- thyl (598 mg, 0.96 mmol), Cs2CO3 (6.2 g, 19.2 mmol), and 1,4-dioxane (80 mL). After three cycles of vacuum/argon flush, the mixture was heated at 115oC for 64 h. It was then 15 cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting e was purified by silica-gel column chromatography eluting with 5:1 petroleum ether/ethyl e to afford 189c as a yellow solid (3.2 g, 75 %). MS: [M+H]+ 444.2. e 189d tert-Butyl 4-(5-Aminopyrazinyl)piperazinecarboxylate 20 189d To a solution of 189c (2.5 g, 5.6 mmol) in methanol (25 mL) was added sodium acetate (0.56 g, 6.8 mmol) and hydroxylamine hydrochloride (0.7 g, 10 mmol). The reaction mixture was stirred for 0.5 h. It was then concentrated under reduced pressure and the residue was purified by column chromatography eluting with 15:1 dichloromethane/methanol to 25 afford 189d(1.3 g, 71%). MS: [M+H]+ 280.3.
Example 189e tert-Butyl 4-(5-(5-Bromomethyloxo-1,2-dihydropyridin- 3-ylamino)pyrazinyl)piperazinecarboxylate 189e A mixture of 189d (1.1 g, 3.94 mmol), 3,5-dibromomethylpyridin-2(1H)-one (1.1 g, 3.94 mmol), ium diacetate (45 mg, 0.20 mmol), (R)-(+)-2,2'-bis(diphenylphosphino)- 30 inaphthyl (245 mg, 0.39 mmol), and Cs2CO3 (2.6 g, 7.9 mmol) in 1,4-dioxane (150 mL) was heated at 120oC for 2 hours. It was then cooled to room temperature and concentrated under reduced pressure. The residue was purified by column chromatography eluting with 30:1 dichloromethane/methanol to afford 189e (900 mg, 54%). MS: [M+H]+ 465.1. 247 Example 189f 5-Bromomethyl(5-(piperazinyl)pyrazin ylamino)pyridin-2(1H)-one 189f A mixture of 189e (1.0 g, 2.2 mmol) and 4.0 M HCl/dioxane (60 mL) was stirred at room temperature for 5 h. It was then concentrated under reduced pressure to afford crude 5 189f as a yellow solid (760 mg, 98%), which was used in the next step without further cation. MS: [M+H]+ 395.1.
Example 189g omethyl(5-(4-(oxetanyl)piperazin yl)pyrazinylamino)pyridine-2(1H)-one 189g 10 A mixture of 189f (740 mg, 2.0 mmol), oxetanone (288 mg, 4.0 mmol), NaBH3CN (315 mg, 5.0 mmol), and zinc chloride (680 mg, 5.0 mmol) in ol (60 mL) was stirred at 50oC for 5 hours. It was then quenched with water and concentrated under reduced pressure. The e was extracted with dichloromethane three times. The combined extract was concentrated under reduced pressure to afford crude 189g as a yellow solid (660 mg, 15 78%), which was used in the next step without further purification. MS: [M+H]+ 423.1.
Example 189h (4-(1-Methyl(5-(4-(oxetanyl)piperazinyl)pyrazin ylamino)oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)pyridinyl)methyl acetate 189h A 50-mL round-bottomed flask equipped with a reflux condenser was charged with 20 189g (180 mg, 0.43 mmol), toxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- l-2(1H)-yl)pyridinylboronic acid 113i (165 mg, 0.43 mmol), Pd(dppf)Cl2 (35 mg, 0.043 mmol), sodium acetate (71 mg, 0.86 mmol), K3PO4 (194 mg, 0.86 mmol), acetonitrile (10 mL), and water (0.5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 3 h. After this time the reaction was filtered and the filtrate was 25 concentrated under reduced pressure. The resulting residue was purified by silica-gel column chromatography eluting with 40:1 dichloromethane/methanol to afford 189h as a yellow solid (100 mg, 34%). MS-ESI: [M+H]+ 680.3. 248 Example 189 2-(3-(hydroxymethyl)(1-methyl(5-(4-(oxetanyl)piperazin yl)pyrazinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-1(2H)-one 189 At room temperature, to a solution of 189h (90 mg, 0.13 mmol) in i-propanol /THF 5 (1:1, 5 mL) and water (0.5 mL) was added lithium hydroxide (126 mg, 2.9 mmol). The reaction e was stirred at 35 oC for 0.5 h. It was then cooled to room temperature and ed. The filtrate was concentrated under reduced pressure and the resulting residue was purified with by reverse-phase prep-HPLC to afford 189 (60 mg, 71%) as yellow solid. MSESI : [M+H]+ 638.3. 1H NMR (500 MHz, CDCl3) δ 8.48 (d, J = 5.0 Hz, 1H), 8.43 (d, J = 2.0 10 Hz, 1H), 7.98 (s, 1H), 7.82 (d, J = 1.5 Hz, 1H), 7.80 (s, 1H), 7.78 (s, 1H), 7.33 (d, J = 5.0 Hz, 1H), 6.89 (s, 1H), .01 (m, 1H), 4.72-4.50 (m, 6H), 4.32-4.30 (m, 1H), 4.15-4.09 (m, 2H), 3.88-3.86 (m, 1H), 3.72 (s, 3H), 3.57-3.49 (m, 5H), 2.61-2.43 (m, 8H), 1.92-1.78 (m, 4H). 15 Example 190a tert-Butyl 3,3-Dimethyl(6-nitropyridinyl)piperazine ylate 190a A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 20 reflux condenser was charged with 5-bromonitropyridine (5.6 g, 28.0 mmol), tert-butyl 3,3-dimethylpiperazinecarboxylate (3.0 g, 14.0 mmol), cesium carbonate (9.1 g, 28 mmol), and 1,4-dioxane (50 mL). After bubbling nitrogen through the resulting solution for 30 min, Binap (870 mg, 1.4 mmol) and tris(dibenzylideneacetone)-dipalladium(0) (1.2 g, 1.4 mmol) were added. The reaction mixture was subjected to three cycles of vacuum/argon 25 flush and stirred at 120 ºC for 24 h. After this time the reaction was cooled to room temperature, ed and the filtrate was partitioned n ethyl acetate (200 mL) and water (50 mL). The s layer was separated and extracted with ethyl acetate (3 × 50 mL).
The combined organic layers were washed with brine (50 mL) and dried over sodium sulfate.
The drying agent was removed by filtration and the filtrate was concentrated under reduced 249 pressure. The residue was purified by silica-gel column chromatography eluting with 5:1 petroleum ethyl e to afford 190a (1.27 g, 27%). LCMS: [M+H]+ 337.2.
Example 190b tert-Butyl 4-(6-Aminopyridinyl)-3,3-dimethylpiperazine carboxylate 190b 5 A 50-mL round-bottomed flask was purged with nitrogen and charged with tert-butyl 3,3-dimethyl(6-nitropyridinyl)piperazinecarboxylate 190a (1100 mg, 3.2 mmol), 10% palladium on carbon (10% wet, 110 mg), and methanol (20 mL). It was then ted, charged with hydrogen gas, and stirred at room temperature for 5 h. The en was evacuated and nitrogen was charged into the flask. The catalyst was removed by filtration 10 through a pad of diatomaceous earth filter agent (CELITE®, Imerys Minerals California, Inc.) and the filtrate was concentrated under reduced pressure to afford 190b (950 mg, 94%).
LCMS: [M+H]+ 307.3 Example 190c tert-Butyl 4-(6-(5-Bromomethyloxo-1,2-dihydropyridin- ino) pyridinyl)-3,3-dimethylpiperazinecarboxylate 190c 15 A 100-mL single-neck round-bottomed flask equipped with a ic stirrer and a reflux condenser was charged with tert-butyl 4-(6-aminopyridin yl)-3,3- dimethylpiperazinecarboxylate 190b (950 mg, 3.1 mmol), 3,5-dibromomethylpyridin- 2(1H)-one (1240 mg, 4.6 mmol), 1,4-dioxane (30 mL), and cesium carbonate (2015 mg, 6.2 mmol). After bubbling nitrogen through the resulting solution for 5 min, Xantphos (179 mg, 20 0.31 mmol) and tris(dibenzylideneacetone)dipalladium(0) (283 mg, 0.31 mmol) were added.
The reaction mixture was subjected to three cycles of /argon flush and heated at reflux for 10 h. After this time the reaction was cooled to room temperature and filtered. The filtrate was partitioned between ethyl acetate (50 mL) and water (10 mL). The aqueous layer was ted and extracted with ethyl acetate (3 × 20 mL). The ed organic layer was 25 washed with brine (30 mL) and dried over sodium sulfate. The drying agent was removed by filtration and the filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 4:1 petroleum ether/ethyl acetate to afford 190c (1.21 g, 79%). LCMS: [M+H]+ 492.1.
Example 190d 5-Bromo(5-(2,2-dimethylpiperazinyl)pyridinylamino)- 30 1-methylpyridin-2(1H)-one 190d To a solution of tert-butyl 4-(6-(5-bromomethyloxo-1,2-dihydropyridin ylamino) pyridinyl)-3,3-dimethylpiperazinecarboxylate 190c (1.19 g, 1.9 mmol) in dichloromethane (20 mL) was added 3M HCl in diethyl ether (15 mL). The on mixture 250 was stirred at room temperature for 4 h. It was then concentrated under reduced pressure to afford 190d (900 mg, 95%). LCMS: [M+H]+ 392.1.
Example 190e 5-Bromo(5-(2,2-dimethyl(oxetanyl)piperazin idinylamino)methylpyridin-2(1H)-one 190e 5 A mixture of 5-bromo(5-(2,2-dimethylpiperazinyl)pyridinylamino)methylpyridin-2 (1H)-one 190d (900 mg, 2.3 mmol), oxetanone (497 mg, 6.9 mmol), NaBH3CN (435 mg, 6.9 mmol), and zinc de (311 mg, 2.3 mmol) in methanol (30 mL) was stirred at 500C for 4 hours. It was then concentrated under reduced pressure. Water (10 mL) was added to the residue and the mixture was extracted with chloroform (3 x 50 mL). The 10 ed organic layer was concentrated under reduced pressure. The residue was purified by silica-gel column-chromatography eluting with 50:1 dichloromethane/methanol to afford 190e (800 mg, 78%). LCMS: [M+H]+ 448.1. 1H NMR (500 MHz, CDCl3) δ 8.65 (d, J = 2.0 Hz, 1H), 8.11 (d, J = 2.5 Hz, 1H), 7.85 (s, 1H), 7.37-7.34 (m, 1H), 6.96 (d, J = 2.5 Hz, 1H), 6.72 (d, J = 8.5 Hz, 1H), 4.69-4.61 (m, 4H), 3.60 (s, 3H), 3.50-3.14 (m, 3H), 2.43-2.17 (m, 15 4H), 1.06 (s, 6H).
Example 190f 2-(4-Chloro(hydroxymethyl)pyridinyl)-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-1(2H)-one 190f O OH N N NaBH4, MeOH N Cl N Cl O N O N 103b 190f OAc OAc N N OH AcCl, DIEA Pin2B2 (5 eq.) N Cl N B DCM a)3, X-phos OH O N AcOK, dioxane O N 65 °C, 15h 190g 190h To a solution of 4-chloro(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- 20 yl)-nicotinaldehyde 103b (1.0 g, 3.0 mmol) in methanol (30 mL) was added sodium borohydride (380 mg, 9.0 mmol) at 10oC. The reaction mixture was stirred for 30 minutes and quenched with water (10 mL). It was then trated under reduced pressure and the residue was dissolved in dichloromethane (50 mL). The mixture was washed with water (10 mL), dried over anhydrous Na2SO4, filtered, and evaporated under d pressure to afford 25 190f as a yellow solid (900 mg, 90%). MS-ESI: [M+H]+ 332. 251 Example 190g (4-Chloro(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl)pyridineyl)methyl Acetate 190g To a e of 2-(4-chloro(hydroxymethyl)pyridinyl)-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-1(2H)-one 190f (900 mg, 2.7 mol) and triethylamine (900 mg, 5 9.0 mol) in dichloromethane (25 mL) was added dropwise acetyl chloride (600 mg, 6.0 mol) while stirring at room ature. The reaction mixture was d for 1 h and concentrated under d re. The residue was ed by silica-gel column chromatography eluting with dichloromethane to afford 190g as white solid (950 mg, 94%). MS-ESI: [M+H]+ 374. 10 Example 190h (4-Chloro(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl)pyridinyl)methyl Acetate 190h A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was d with (4-chloro(1-oxo-3,4,6,7,8,9-hexahydro-pyrazino[1,2- a]indol-2(1H)-yl)pyridinyl)methyl acetate 190g (950 mg, 2.5 mmol), 15 Pin2B2(4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (1.6 g, 2.0 eq., 5 mmol), Pd2(dba)3 (230 mg, 0.1 eq., 0.25 mmol), X-phos (232 mg, 0.2 eq., 0.50 mmol), potassium acetate (735 mg, 3 eq., 7.5 mmol), and dioxane (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at 65ºC for 15 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was 20 washed with 3:1 petroleum ether/ethyl acetate to afford 190h as yellow solid (950 mg, 87%).
MS-ESI: [M+H]+ 383.
Example 190i (4-(5-(5-(2,2-Dimethyl(oxetanyl)piperazinyl)pyridin ylamino)methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl Acetate 190i 25 A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 5-bromo(5-(2,2-dimethyl(oxetanyl)piperazin yl)pyridinylamino)methylpyridin-2(1H)-one 190e (200 mg, 1.0 eq., 0.45 mmol), (2-(1- 252 oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)(4,4,5,5-tetramethyl-1,3,2- dioxaborolanyl)pyridinyl)methyl acetate 190h (345 mg, 2 eq., 0.90 mmol), PdCl2(dppf) (36 mg, 0.1 eq., 0.045 mmol), K3PO4 (191 mg, 2 eq., 0.90 mmol), sodium acetate (74 mg, 2.0 eq., 0.90 mmol), itrile (15 mL), and water (0.1 mL). After three cycles of 5 vacuum/argon flush, the mixture was stirred at 90oC for 2 h. It was then cooled to room ature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 50:1 dichloromethane/ethanol to afford 190i (100 mg, 31%) as yellow solid. MS-ESI: [M+H]+ 707.4. 10 Example 190 2-[4-[5-[[5-[2,2-dimethyl(oxetanyl)piperazinyl] pyridyl]amino]methyloxopyridyl](hydroxymethyl)pyridyl]-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indolone 190 A 50-mL -neck round-bottomed flask equipped with a magnetic stirrer was charged with (4-(5-(5-(2,2-dimethyl(oxetanyl)piperazinyl)pyridinylamino) 15 methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl)pyridinyl)methyl acetate 190i (100 mg, 1 eq., 0.14 mmol), lithium hydroxide(54 mg, 10 eq., 1.4 mmol), i-propanol (3 mL), THF (3 mL) and water (2 mL). The mixture was stirred at 30oC for 1 h. It was then filtered and the filtrate was concentrated under d pressure. The residue was purified by reverse-phase prep-HPLC to afford 190 as a white solid 20 (43 mg, 46%). MS-ESI: [M+H]+ 665.3. 1H NMR (500 MHz, DMSO-d6) δ 8.69 (d, J = 2.0 Hz, 1H), 8.60 (s, 1H), 8.48 (d, J = 5.0 Hz, 1H), 7.93 (d, J = 2.0 Hz, 1H), 7.51(d, J = 1.5 Hz, 1H), 7.42-7.40 (m, 1H), 7.35 (d, J = 5.0 Hz, 1H), 7.24 (d, J = 8.5 Hz, 1H), 6.58 (s, 1H), 4.98 (brs, 1H), 4.54 (t, J = 6.0 Hz, 2H), 4.46-4.38 (m, 4H), 4.25-3.85 (m, 4H), 3.60 (s, 3H), 3.38- 3.35 (m, 1H), 3.03-2.54 (m, 4H), 2.47 (t, J = 6.0 Hz, 2H), 2.32-2.12 (m, 4H), 1.79-1.67 (m, 25 4H), 0.97 (s, 6H) Example 191a N-tert-Butyl-4,5,6,7-tetrahydrobenzo[b]thiophene carboxamide 191a A mixture of 4,5,6,7-tetrahydrobenzo[b]thiophenecarboxylic acid (500 g, 2.75 mol, 1.0 equiv) and thionyl chloride (655 g, 5.5 mol, 2.0 equiv) was heated under reflux for 3 h. 30 Excess l chloride was removed by distillation under reduced pressure. The e was taken up in dichloromethane (1.0 L) and a solution of tert-butylamine (402 g, 5.5 mol, 2.0 equiv) in dichloromethane (500 mL) was added with stirring while the temperature of the mixture being kept below 10oC. The resulting solution was stirred at 25oC for 16 h. Most of the solvent was d under reduced pressure. The residue was chilled in an ice-bath and 253 2M KOH solution was introduced slowly to adjust the pH to 11 with stirring. The suspension was filtered and the solid collected, washed three times with water, and dried in vacuum to afford 191a as a white solid (580 g, 80%, over two steps). MS: [M+H]+ 238. 1H NMR (500 MHz, CDCl3) δ 7.02 (s, 1H), 5.77 (s, 1H), 2.65 (t, J = 6.0 Hz, 1H), 2.47 (t, J = 6.0 Hz, 1H), 5 1.74-1.70 (m, 4H), 1.35 (s, 9H).
Example 191c N-tert-Butyl(diazenylmethyl)-4,5,6,7-tetrahydrobenzo- [b]thiophenecarboxamide 191c A solution of 191a (100 g, 0.42 mol, 1.0 equiv) in THF (500 mL) was slowly added to n-butyl lithium (672 mL, 2.5M in THF, 1.68 mol , 4.0 equiv) at -78oC under argon protection. 10 The mixture was stirred for 2 h. N,N-Dimethylformamide (306 g, 4.2 mol, 10.0 equiv) was added to the mixture while the temperature being sustained at -78oC. After another 2.0 h, the reaction mixture was quenched by addition of methanol (500 mL) at -78oC. It was stirred for 0.50 h at room temperature to afford 191b in situ. Then 80% aqueous hydrazine hydrate (131 g, 2.1 mol) was added and the mixture was ed at 65oC overnight. The c solvent 15 was removed under reduced pressure. The e was filtered and the resulting yellow solid was washed with water. The solid was dried in vacuum to afford 191c, which was used for the next step without further purification. MS: [M+H]+ 280.
Example 191d 8-Thia-4,5-diazatricyclo[7.4.0.02,7]trideca-(9),2(7),3-trien one 191d 20 A mixture of N-tert-butyl(diazenylmethyl)-4,5,6,7-tetrahydrobenzo[b]thiophene carboxamide 191c (40 g, 144 mmol) in H2SO4 (30% s, 3 L) was refluxed at 105oC for 24 h. It was then filtered and the filtrate was extracted with dichloromethane (3ⅹ1 L). The combined extract was dried over Na2SO4 and evaporated under d re. The residue 254 was purified by silica-gel column tography eluting with 100:1 dichloromethane/methanol to afford 191d as a white solid (9.0 g, 31%). MS: [M+H]+ 207. 1H NMR (500 MHz, CDCl3) δ 8.15 (s, 1H), 2.96-2.94 (m, 2H), 2.86-2.84 (m, 2H),1.96-1.94 (m, 4H). 5 Example 191e (3S)-tert-Butyl 3-Methyl(6-nitropyridinyl)piperazine ylate 191e Following the procedure described for compound 101g and starting with (3S)-tertbutyl 3-methylpiperazinecarboxylate (10.0 g, 50 mmol) and onitropyridine (10.5 g, 50 mmol) afforded 191e as a yellow solid (8.05 g, 50%). MS-ESI: [M+H]+ 323 10 Example 191f (3S)-tert-Butyl 4-(6-Aminopyridinyl)methylpiperazine carboxylate 191f Following the procedure described for compound 101h and starting with (3S)-tertbutyl 3-methyl(6-nitropyridinyl)piperazinecarboxylate 191e (5.8 g, 18 mmol) afforded 191f as a brown solid (4.9 g, 93%). MS-ESI: [M+H]+ 293 15 Example 191g (3S)-tert-Butyl 4-(6-(5-Bromomethyloxo-1,2-dihydropyridin ylamino) neyl)methylpiperazinecarboxylate 191g Following the procedure described for compound 101i and ng with (3S)-tertbutylmethyl (6-nitropyridinyl)piperazinecarboxylate 191f (4.0 g, 13.7 mmol) and 3,5-dibromomethylpyridin-2(1H)-one (5.5 g, 20.6 mmol) afforded 191g as a yellow solid 20 (5.4 g, 83%). MS-ESI: [M+H]+ 478 Example 191h (3S)Bromomethyl(5-(2-methylpiperazinyl)pyridin- 2-ylamino)pyridine-2(1H)-one 191h 255 Following the procedure described for nd 101j and starting with (3S)-tertbutyl 4-(6-(5-bromomethyloxo-1,2-dihydropyridinylamino)pyridineyl)methylpiperazinecarboxylate 191g (3.1 g, 6.5 mmol) afforded 191h as a yellow solid (2.3 g, 94%). MS-ESI: [M+H]+ 378. 5 Example 191i (S)Bromomethyl(5-(2-methyl(oxetanyl)piperazin yl)pyridinylamino)pyridin-2(1H)-one 191i A mixture of (S)bromomethyl(5-(2-methylpiperazinyl)pyridin ylamino)pyridin-2(1H)-one 191h (40.0 g, 106 mmol), one (11.4 g, 159 mmol), NaBH3CN (10.0 g, 159 mmol), and zinc chloride (21.3 g, 159 mmol) in methanol (700 mL) 10 was stirred at 50oC for 5 hours. The mixture was added to water (100 mL) and concentrated under reduced pressure. The residue was extracted with romethane (200 mL × 3). The combined organic layer was concentrated under reduced pressure and the residue was purified by silica-gel column chromatography eluting with 40:1 dichloromethane nol to afford 191i (35 g, 73%). MS: [M+H]+ 434. 15 Example 191j (3S)Methyl(5-(2-methyl(oxetanyl)piperazinyl)- pyridinylamino) (4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 191j O O N N N N O O O N NH B B O N NH O O O 191h O ZnCl2, NaCNBH3, Pd2(dba)3, X-Phos, O N B MeOH, 50 °C, 2 h KOAc, dixoane, N Br 70°C, 2 h O 191i 191j A 100-mL single-neck round-bottomed flask equipped with a magnetic r and a reflux condenser was charged with (S)-tert-butyl(6-(5-bromomethyloxo-1,2- 20 dihydropyridinylamino)pyridineyl)methylpiperazinecarboxylate 191i (1.0 g, 1.0 eq., 2.3 mmol), Pin2B2 (1.46 g, 2.50 eq., 5.75 mmol), Pd2(dba)3 (105 mg, 0.05 eq., 0.125 mmol), X-Phos (93 mg, 0.1 eq., 0.23 mmol), potassium acetate (676 mg, 3.0 eq., 6.9 mmol), and dioxane (50 mL). After three cycles of vacuum/argon flush, the mixture was heated at 90ºC for 4 h. It was then cooled to room temperature and filtered. The filtrate was 25 trated under reduced pressure and the resulting residue was washed with 3:1 petroleum ether/ethyl acetate (80 mL) to afford 191j as yellow solid (1.0 g, 90%). MS: [M+H]+ 482. 256 Example 191k 4-[1-Methyl({5-[(2S)methyl(oxetanyl)piperazin yl]pyridinyl}amino)oxo-1,6-dihydropyridinyl]{6-oxothia-4,5-diazatricyclo [7.4.0.02,7]tri-deca-1(9),2(7),3-trienyl}pyridinecarbaldehyde 191k 5 A 50-mL round-bottomed flask ed with a reflux condenser was d with (S)methyl(5-(2-methyl(oxetanyl)piperazinyl)pyridinylamino)(4,4,5,5- tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 191j (168 mg, 0.35 mmol), 4-chloro- 2-{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridine dehyde 124a (121 mg, 0.35 mmol), K3PO4 (148 mg, 0.70 mmol), 1,1’- 10 bis(diphenylphosphino)ferrocenedichloropalladium(II) (13 mg, 0.0175 mmol), sodium e trihydrate (95 mg, 0.70 mmol), water (6 drops), and acetonitrile (10 mL). After bubbling nitrogen through the mixture for 30 minutes, it was heated at 100°C under N2 protection for 1h. Analysis of reaction mixture by LCMS showed completed conversion to the desired product. The reaction mixture was cooled to room temperature and filtered. The 15 filtrate was concentrated under reduced pressure. The residue was d with dichloromethane (30 mL) and water (30 mL). The aqueous layer was separated and extracted with dichloromethane (3 × 20 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The dark residue was purified by silica-gel column chromatography eluting with dichloromethane /methanol (80/1 to 30/1) to afford 20 191k (118 mg, 51%) as yellow solid. MS-ESI: [M+H]+ 665 Example 191 5-[3-(Hydroxymethyl)[1-methyl({5-[(2S)methyl(oxetan yl)pipera-zinyl]pyridinyl}amino)oxo-1,6-dihydropyridinyl]pyridinyl]thia- 4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienone 191 To a solution of 4-[1-methyl({5-[(2S)methyl(oxetanyl)piperazin 25 idinyl}amino)oxo-1,6-dihydropyridinyl]{6-oxothia-4,5-diazatricyclo- [7.4.0.02,7]-trideca-1(9),2(7),3-trienyl}pyridinecarbaldehyde 191k (118 mg, 0.18 mmol) 257 in methanol/ dichloromethane (10/10 mL) was added NaBH4 (21 mg, 0.54 mmol) at room temperature. After the reaction was stirred for 1 h, LCMS indicated the reaction was completed. Then the e was poured into water (20 mL) and concentrated under reduced re. The residue was extracted with dichloromethane (20 mL × 3). The combined 5 organic layer was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue solid was purified by reverse-phase prep-HPLC to afford 191 (71 mg, 60%) as white solid. MS-ESI: [M+H]+ 667. 1H NMR (500 MHz, DMSO-d6) δ 8.64 (d, J = 2.5 Hz, 1H), 8.58 (d, J = 5.0 Hz, 1H), 8.48-8.46 (m, 2H), 7.86 (d, J = 3.0 Hz, 1H), 7.54 (d, J = 5.5 Hz, 1H), 7.48 (d, J = 2.0 Hz, 1H), 7.37 (dd, J = 3.0, 9.0 Hz, 1H), 7.25 10 (d, J = 9.5 Hz, 1H), 4.86-4.85 (m, 1H), 4.58-4.55 (m, 2H), 4.48-4.46 (m, 2H), 4.42-4.40 (m, 2H), 3.65-3.64 (m, 1H), 3.61 (s, 3H), 3.41-3.99 (m, 1H), 3.05-3.04 (m, 1H), 2.97-2.95 (m, 3H), 2.87-2.86 (m, 2H), 2.52-2.51 (m, 1H), 2.34-2.32 (m, 2H), 2.21-2.20 (m, 1H), 1.89-1.87 (m, 4H), 0.94 (d, J = 6.0 Hz, 3H). e 192a Methyl 1-Formyl-5,6,7,8-tetrahydroindolizinecarboxylate 192a 15 A 100-mL round-bottomed flask equipped with a magnetic stirrer was purged with nitrogen and charged with anhydrous dichloroethane (10 mL) and anhydrous DMF (0.7 mL, 9.0 mmol). The reaction mixture was cooled to 0oC and phosphorus oride (0.7 mL, 7.3 mmol) was added over a period of 2 min, while maintaining the reaction temperature between 0oC and 10oC. The cooling bath was removed and the reaction was stirred at room 20 temperature for 1 hour. A solution of methyl 5,6,7,8-tetrahydroindolizinecarboxylate 112a (1.0 g, 5.6 mmol) in acetonitrile (10 mL) was added and the e was stirred at room temperature for additional 3 hours. After this time, the t was concentrated under reduced pressure and the oily residue was taken up with saturated aqueous NaHCO3 (20 mL).
The aqueous layer was ted with ethyl acetate (3 x 70 mL). The combined organic layer 25 was washed with water (20 mL), dried over Na2SO4 and ated under reduced pressure.
The residue was purified by silica-gel column chromatography eluting with 1:1 ethyl acetate/petroleum ether to afford 192a as a white solid (406 mg, 33%). MS: (M+H)+ 208.3. 1H NMR (500 MHz, DMSO) δ 10.29 (s, 1H), 7.43 (s, 1H), 3.99 (t, J = 6.0 Hz, 2H), 3.76 (s, 3H), 2.95 (t, J = 6.5 Hz, 2H), 1.90-1.85 (m, 2H), 1.78-1.74 (m, 2H). 30 e 192b 7,8,9,10-Tetrahydropyridazino[4,5-a]indolizin-4(3H)-one 192b 258 A 100 mL single-neck round-bottomed flask was charged with hydrazinium hydroxide (20 mL), methyl 1-formyl-5,6,7,8-tetrahydroindolizinecarboxylate 192a (2.5 g, 12.0 mmol). The reaction mixture was heated at 100 oC for 4 hours. After this time the 5 reaction was cooled to room temperature and filtered to afford 192b as a yellow solid (1.9 g, 83%). MS: (M+H)+ 190.3. 1H NMR (500 MHz, DMSO) δ 11.44 (s, 1H), 8.03 (s, 1H), 7.42 (s, 1H), 4.18 (t, J = 6.0 Hz, 2H), 2.96 (t, J = 6.5 Hz, 2H), 1.98-1.93 (m, 2H), 1.87-1.82 (m, 2H).
Example 192c 4-Chloro(4-oxo-7,8,9,10-tetrahydropyridazino[4,5- a]indolizin-3(4H)-yl)nicotinaldehyde 192c 10 A 100-mL single-neck bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 1,4-dioxane (50 mL), ium carbonate (1.5 g, 10.6 mmol), 10-tetrahydropyridazino[4,5-a]indolizin-4(3H)-one 192b (1.0 g, 5.3 mmol), and 2-bromochloronicotinaldehyde (3.5 g, 15.9 mmol). After bubbling nitrogen h the resulting mixture for 30 minutes, copper(I) bromide (75.0 mg, 0.53 mmol) and sarcosine 15 (47.0 mg, 0.53 mmol) were added, and the reaction e was heated at 95oC for 12 h.
After this time the reaction was cooled to room temperature and filtered. The filtrate was partitioned between methylene chloride (60 mL) and water (40 mL). The aqueous layer was separated and extracted with methylene chloride (3 x 70 mL). The combined organic layer was washed with brine (30 mL) and dried over sodium sulfate. The drying agent was 20 removed by filtration and the te was concentrated under reduced pressure. The residue was ed by silica-gel column chromatography eluting with 10:1 ethyl acetate/petroleum ether to afford 192c as a brown solid (521 mg, 30%). MS-ESI: [M+H]+ 329.2.
Example 192d (S)(1-Methyl(5-(2-methyl(oxetanyl)piperazin yl)pyridinylamino)oxo-1,6-dihydropyridinyl)(4-oxo-7,8,9,10- 25 tetrahydropyridazino[4,5-a]indolizin-3(4H)-yl)nicotinaldehyde 192d 259 A 100-mL round-bottomed flask equipped with a reflux condenser was charged with 4-chloro(4-oxo-7,8,9,10-tetrahydropyridazino[4,5-a]indolizin-3(4H)-yl)nicotinaldehyde 192c (196 mg, 0.60 mmol), (S)methyl(5-(2-methyl(oxetanyl)-piperazin 5 yl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 191j (290 mg, 0.60 mmol), sodium acetate (100 mg, 1.2 mmol), K3PO4 (320 mg, 1.2 mmol), PdCl2(dppf) (50 mg, 0.060 mmol), acetonitrile (25 mL), and water (1 mL). After bubbling nitrogen through the mixture for 30 minutes, it was heated at 100 oC for 3 hours under N2 protection. The reaction mixture was cooled to room temperature and evaporated under 10 reduced pressure. The residue was purified by silica-gel column tography eluting with 20:1 methylene de/methanol to afford 192d 173 mg, 44%) as a brown solid. MS-ESI: [M+H]+ 648.4.
Example 192 3-[3-(hydroxymethyl)[1-methyl[[5-[(2S)methyl(oxetan yl)piperazinyl]pyridyl]amino]oxopyridyl]pyridyl]-7,8,9,10- 15 ydropyridazino[4,5-a]indolizinone 192 To a solution of 192d (160 mg, 0.25 mmol) in MeOH (20 mL) was added NaBH4 (28 mg, 0.75 mmol). The e was stirred at 20oC for 2 h and evaporated under reduced pressure. The e was purified by reverse-phase prep-HPLC to afford 192 (97 mg, 60%) as a yellow solid. MS-ESI: [M+H]+ 650.4. 1H NMR (500 MHz, DMSO-d6) δ 8.65 (d, J = 2.5 20 Hz, 1H), 8.54 (d, J = 4.5 Hz, 1H), 8.45 (s, 1H), 8.25 (s, 1H), 7.86 (d, J = 2.5 Hz, 1H), 7.62 (s, 1H), 7.51 (d, J = 2.5 Hz, 1H), 7.47 (d, J = 5.0 Hz, 1H), 7.38-7.36 (m, 1H), 7.25 (d, J = 9.5 Hz, 1H), 4.66 (s, 1H), 4.57-4.54 (m, 2H), 4.48-4.46 (m, 1H), 4.43-4.41 (m, 1H), 4.33 (s, 2H), 4.25-4.21 (m, 2H), 3.69-3.66 (m, 1H), 3.60 (s, 3H), .37 (m, 1H), 3.11-3.07 (m, 1H), 3.06-3.04 (m, 2H), 2.97-2.92 (m, 1H), 2.55-2.53 (m, 1H), 2.35-2.29 (m, 2H), 2.19-2.17 (m, 25 1H), 2.04-1.96 (m, 2H), 1.93-1.85 (m, 2H), 0.93 (d, J = 6.5 Hz, 3H). 260 Example 193a Methyl 2-(Hydroxy(pyridinyl)methyl)acrylate 193a A 250-mL -neck round-bottomed flask was charged with chloroform (100 mL), picolinaldehyde (10.7 g, 0.10 mol), methyl acrylate (8.60 g, 0.10 mol), and 1,4-diaza- 5 bicyclo[2.2.2]octane (0.560 g, 5.00 mmol). The reaction e stirred at room temperature for 48 h. After this time the reaction was concentrated under reduced pressure and the residue was purified by silica-gel column chromatography eluting with 3:1 petroleum ether/ethyl acetate to afford 193a as dark yellow oil (11.6 g, 60%). MS-ESI: (M+H)+ 194.2. 1H NMR (500 MHz, CDCl3) δ 8.54 (d, J = 5.0 Hz, 1H), .66 (m, 1H), 7.42 (d, J = 8.0 Hz, 1H), 10 7.22-7.20 (m, 1H), 6.36 (s, 1H), 5.97 (s, 1H), 5.62 (s, 1H), 4.85 (s, 1H), 3.74 (s, 3H).
Example 193b Methyl Indolizinecarboxylate 193b A 250-mL -neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with acetic anhydride (80 mL) and 193a (6.68 g, 34.6 mmol).
The reaction mixture was heated at reflux under nitrogen for 4 h. After this time the on 15 was cooled to room temperature, poured onto the mixture of ice (100 g) and aqueous saturated sodium bicarbonate (200 mL), and stirred for 1 h. The resulting solution was neutralized with saturated aqueous sodium bicarbonate and extracted with methylene de (3 × 200 mL). The combined organic extract was dried over sodium sulfate and concentrated under reduced re. The residue was purified by silica-gel column chromatography 20 eluting with 10:1 petroleum ether/ethyl acetate (10:1) to afford 193b as a white solid (2.1 g, 35%). MS-ESI: (M+H)+ 176.2. 1H NMR (500 MHz, CDCl3) δ 7.86-7.84 (m, 1H), 7.79 (d, J = 1.0 Hz, 1H), 7.36-7.34 (m, 1H), 6.82 (s, 1H), 6.70-6.66 (m, 1H), 6.55-6.51 (m, 1H), 3.88 (s, 3H).
Example 193c Methyl 5,6,7,8-Tetrahydroindolizinecarboxylate 193c 261 A 250-mL round-bottomed flask was purged with nitrogen and charged with 193b (2.0 g, 11.4 mmol), 10% palladium on carbon (50% wet, 200 mg), and methanol (50 mL). It was evacuated, charged with hydrogen gas, and stirred under 5 atm hydrogen at room temperature for 8 h. The hydrogen was then evacuated and nitrogen was charged into the 5 flask. The catalyst was d by filtration through a pad of CELITE® and the filtrate concentrated under reduced re to afford 193c as a white solid (1.1 g, 81%). MS-ESI: [M+H]+ 180.3. 1H NMR (500 MHz, DMSO-d6) δ 7.25 (d, J = 2.0 Hz, 1H), 6.09 (s, 1H), 3.93 (t, J = 6.0 Hz, 2H), 3.66 (s, 3H), 2.67 (t, J = 6.0 Hz, 2H), 1.87-1.83 (m, 2H), 1.75-1.70 (m, 2H). 10 Example 193d Methyl 3-Formyl-5,6,7,8-tetrahydroindolizinecarboxylate 193d A 100-mL round-bottomed flask ed with a magnetic stirrer was purged with nitrogen and charged with anhydrous dichloroethane (20 mL) and anhydrous DMF (0.70 mL, 9.0 mmol). To the mixture at 0oC was added phosphorus oxychloride (0.70 mL, 7.3 mmol) 15 over a period of 2 min, while maintaining the on temperature between 0oC and 10oC.
The cooling bath was removed and the reaction was stirred at room temperature for 1 hour. A solution of 193c (1.0 g, 5.6 mmol) in acetonitrile (10 mL) was added and the reaction mixture was stirred at room ature for 3 hours. After this time, it was concentrated under reduced pressure. The oily residue was taken up with saturated aqueous NaHCO3 (20 mL) 20 and extracted with ethyl acetate (3 x 50 mL). The combined organic layer was washed with water (50 mL), dried over Na2SO4, and evaporated under reduced pressure. The residue was purified by silica-gel column chromatography g with 1:5 ethyl acetate/petroleum ether to afford 193d as a white solid (703 mg, 58%). MS-ESI: (M+H)+ 208.3. 1H NMR (500 MHz, 6) δ 10.14 (s, 1H), 6.40 (s, 1H), 4.27 (t, J = 6.0 Hz, 2H), 3.78 (s, 3H), 2.78 (t, J = 25 6.0 Hz, 2H), 1.94-1.85 (m, 2H), 1.78-1.69 (m, 2H).
Example 193e 9-Tetrahydropyridazino[4,5-b]indolizin-1(2H)-one 193e A 100-mL single-neck round-bottomed flask equipped with a reflux condenser was charged with 193d (600 mg, 2.9 mmol) and hydrazine hydrate (20 mL). The reaction mixture was heated at 100oC for 4 hours. After this time the reaction was cooled to room temperature 30 and filtered to afford 193e as a yellow solid (413 mg, 75%). MS-ESI: (M+H)+ 190.1. 1H NMR (500 MHz, DMSO-d6) δ 12.17 (s, 1H), 8.24 (s, 1H), 6.33 (s, 1H), 4.16 (t, J = 6.0 Hz, 2H), 2.88 (t, J = 6.5 Hz, 2H), 2.00-1.96 (m, 2H), 1.84-1.79 (m, 2H). e 193f 4-Chloro(1-oxo-6,7,8,9-tetrahydropyridazino[4,5- b]indolizin-2(1H)-yl)nicotinaldehyde 193f 262 A 100-mL single-neck round-bottomed flask equipped with a reflux condenser was charged with 1,4-dioxane (40 mL), 193e (800 mg, 3.6 mmol), 2-bromo chloronicotinaldehyde (2.8 g, 12.6 mmol), and potassium carbonate (1.2 g, 8.4 mmol). After bubbling nitrogen through the resulting mixture for 30 minutes, copper(I) iodide (800 mg, 4.2 5 mmol) and 4,7-dimethoxy-1,10-phenanthroline (1.0 g, 4.2 mmol) were added, and the reaction mixture was heated at 90oC for 12 h. After this time the reaction was cooled to room temperature and filtered. The filtrate was ioned between methylene chloride (60 mL) and water (40 mL). The s layer was separated and ted with methylene chloride (3 x 40 mL). The combined organic layer was washed with brine (30 mL) and dried over 10 sodium sulfate. The drying agent was removed by filtration and the filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with1:1 ethyl e/petroleum ether to afford 193f as a brown solid (513 mg, yield 37%). MS-ESI: [M+H] + 329.1. e 193g (S)(1-Methyl(5-(2-methyl(oxetanyl)piperazinyl)- 15 pyridinyl-amino)oxo-1,6-dihydropyridinyl)(1-oxo-6,7,8,9 tetrahydropyridazino [4,5-b]indolizin-2(1H)-yl)nicotinaldehyde 193g A 100-mL round-bottomed flask equipped with a reflux condenser was charged with 193f (200 mg, 0.61 mmol), 191j (293 mg, 0.60 mmol), sodium acetate (98 mg, 1.2 mmol), 20 K3PO4 (254 mg, 1.2 mmol), PdCl2(dppf) (50 mg, 0.060 mmol), itrile (25 mL), and water (1 mL). After bubbling nitrogen through the mixture for 30 minutes, it was heated at 263 100oC for 3 hours. The mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica-gel column chromatography eluting with 25:1 methylene chloride/methanol to afford 193g (206 mg, 53%) as a brown solid. MS-ESI: [M+H] + 648.3. 5 Example 193 2-[3-(hydroxymethyl)[1-methyl[[5-[(2S)methyl(oxetan yl)piperazinyl]pyridyl]amino]oxopyridyl]pyridyl]-6,7,8,9- ydropyridazino[4,5-b]indolizinone 193 To a solution of 193g (180 mg, 0.28 mmol) in methanol (20 mL) was added NaBH4 (32 mg, 0.84 mmol). The mixture was stirred at 20oC for 2 h and quenched with water. It was 10 then evaporated under reduced pressure and the residue was purified by reverse-phase prep- HPLC to afford 193 (140 mg, 78%) as an ite solid. MS-ESI: [M+H] + 650.4. 1H NMR (500 MHz, DMSO-d6) δ 8.67 (d, J = 2.0 Hz, 1H), 8.55 (d, J = 5.5 Hz, 1H), 8.47 (s, 1H), 8.48 (s, 1H), 7.85 (d, J = 2.5 Hz, 1H), .50 (m, 2H), 7.38-7.36 (m, 1H), 7.25 (d, J = 8.5 Hz, 1H), 6.49 (s, 1H), 4.72 (t, J = 5.0 Hz, 1H), 4.57-4.54 (m, 2H), 4.47 (t, J = 6.0 Hz, 1H), 4.41 (t, 15 J = 6.0 Hz, 1H), 4.33-4.29 (m, 2H), 4.28-4.25 (m, 2H), 3.71-3.65 (m, 1H), 3.60 (s, 3H), 3.41- 3.77 (m, 1H), 3.10-3.08 (m, 1H), .90 (m, 3H), 2.57-2.52 (m, 1H), 2.35-2.30 (m, 2H), 2.18-2.16 (m, 1H), 2.06-2.0 (m, 2H), 1.89-1.82 (m, 2H), 0.93 (d, J = 6.0 Hz, 3H).
Example 194a 1-Methyl(pyrazinylamino)(4,4,5,5-tetramethyl-1,3,2- dioxaborolanyl)pyridin-2(1H)-one 194a 20 A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with 5-bromomethyl(pyrazinylamino)pyridin-2(1H)- one 142a (600 mg, 2.0 mmol), Pin2B2 (2.54 g, 10 mmol), Pd2(dba)3 (100 mg, 0.10 mmol), X- phos (100 mg, 0.20 mmol), potassium acetate (600 mg, 6.0 mmol), and e (80 mL).
After three cycles of vacuum/argon flush, the mixture was heated at 65 ºC for 16 h. It was 25 then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was washed with petroleum ether to afford 194a as a yellow solid (crude product) (1.0 g, LCMS purity: 70%). MS-ESI: [M+H]+ 329.4.
Example 194b 4-{1-Methyloxo[(pyrazinyl)amino]-1,6- dihydropyridinyl}{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trien 30 yl}pyridinecarbaldehyde 194b 264 A 100-mL single-neck round-bottomed flask equipped with a reflux condenser was charged with 4 -chloro{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trien- 5-yl}pyridinecarbaldehyde 124a (345 mg, 1.0 mmol), 194a (659 mg, 2.0 mmol), 5 Pd(dppf)Cl2 (50 mg, 0.050 mmol), K3PO4 (450 mg, 2.0 mmol), sodium acetate trihydrate (300 mg, 2.0 mmol), water (6 drops) and, acetonitrile (40 mL). The system was evacuated and refilled with N2. The reaction mixture was heated at 100ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 25:1 10 dichloromethane/methanol to afford 194b (250 mg, 49%) as a yellow brown solid. MS-ESI: [M+H]+ 512.3.
Example 194 3-[3-(hydroxymethyl)[1-methyloxo(pyrazinylamino) pyridyl]pyridyl]-6,7,8,9-tetrahydrobenzothiopheno[2,3-d]pyridazinone 194 A mixture of 194b (200 mg, 0.4 mmol) and NaBH4 (48 mg, 1.2 mmol) in ol 15 (20 mL) was stirred at 30oC for 2 h. The mixture was quenched with water (5 mL) and concentrated under reduced re. The residue was extracted with ethyl acetate (3 x 10 mL). The ed with ethyl acetate extract was trated under reduced re and the residue was purified by reverse-phase prep-HPLC to afford 194 (60 mg, 30%) as a white solid. MS-ESI: [M+H]+ 514.2. 1H NMR (500 MHz, CDCl3) δ 8.78 (d, J = 2.0 Hz, 1H), 8.69 20 (d, J = 5.0 Hz, 1H), 8.31 (s, 1H), 8.29 (s, 1H), 8.17 (s, 1H), 8.13 (s, 1H), 8.04 (d, J = 2.5 Hz, 265 1H), 7.83 (d, J = 2.5 Hz, 1H), 7.56 (d, J = 5 Hz, 1H), 4.44-4.37 (m, 3H), 3.76 (s, 3H), 3.01- 2.99 (m, 2H), 2.88-2.86 (m, 2H), 2.01-1.96 (m, 4H).
Example 195a Ethyl 4,5,6,7-Tetrahydro-1H-indolecarboxylate 195a OEt Br O O NaOH, THF, NaN3, DMSO O OEt EtOH, rt, O/N O N O 105 oC, 4 h N O NaH, DMF, 80oC Cl H EtO overnight OEt 195a 195b OH HATU, Et3N, DMAP, NH2 O N O DMF, NH3H2O, rt., O/N N O AcOH, 110oC N NH EtO EtO 1 h OEt OEt 195c 195d 195e 5 To a mixture of ethyl 3-(2-chlorocyclohexenyl)acrylate (21.4 g, 100 mmol) in DMSO (100 mL) was added sodium azide (9.75 g, 150 mmol). The on mixture was heated at 105oC for 4 h. After cooling to room temperature, the mixture was poured into ice water. The resulting precipitate was collected by filtration to afford 195a (18.0 g, 93.3%).
MS-ESI: [M+H]+ 194. 10 Example 195b Ethyl 1-(2,2-Diethoxyethyl)-4,5,6,7-tetrahydro-1H-indole carboxylate 195b To a suspension of NaH (1.44 g, 60.2 mmol) in N,N-dimethylformamide (DMF)(30 mL) was slowly added 195a ( 5.80 g, 30.1 mmol) at 0oC. The resulting mixture was stirred at room temperature for 0.5 h, ed by the addition of 2-bromo-1,1-diethoxyethane (11.9 g, 15 60.2 mmol). The reaction was heated at 70oC for 30 h and quenched with water (100 mL).
The e was then extracted with ethyl acetate (3 x 100 mL). The combined organic phase was concentrated under reduced pressure and the e was purified with silica-gel column chromatography eluting with 40:1 petroleum ether/ethyl acetate to 195b (4.7 g, 51%). MSESI : [M-EtOH+H]+ 264. 1H NMR (500 MHz, 6) δ 6.65 (s, 1H), 4.59 (t, J =5.0 Hz, 20 1H), 4.17-4.16 (m, 4H), 3.59-3.57 (m, 2H), 3.27-3.26 (m, 2H), 2.61 (t, J = 6.0 Hz, 2H), 2.51 (t, J = 6.0 Hz , 2H), 1.73-1.71 (m, 2H), 1.63-1.61 (m, 2H), 1.25 (t, J = 7.0 Hz, 3 H), 1.02 (t, J = 7.0 Hz, 6H). e 195c 1-(2,2-Diethoxyethyl)-4,5,6,7-tetrahydro-1H-indole carboxylic Acid 195c 266 To a mixture of 195b (4.7 g, 15.2 mmol) in a mixed t of ethanol (20 mL), tetrahydrofuran (20 mL), and water (30 mL) was added sodium hydroxide (3.0 g, 75.0 mmol).
The reaction was heated at 75oC for two days and concentrated under reduced pressure. The residue was suspended in water and neutralized with diluted aqueous citric acid solution. The 5 mixture was extracted with ethyl acetate (3 X 100 mL) and the combined c phase was concentrated under reduced pressure to afford 195c (3.32 g, 78%). MS-ESI: [M-EtOH+H]+ 236.
Example 195d 1-(2,2-Diethoxyethyl)-4,5,6,7-tetrahydro-1H-indole carboxamide 195d 10 To a mixture of 195c (2.8 g, 10.0 mmol) in N,N-dimethylformamide (30 mL) was added O-(7-azabenzotriazolyl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HATU) (5.7 g, 15.0 mmol), Et3N (1.5 g, 15.0 mmol), and DMAP (128 mg, 1.0 mmol). The reaction mixture was stirred at room temperature for overnight. ted ammonium hydroxide (30 mL) was added and the resulting mixture was further stirred for 2 h. It was 15 then d with water (100 mL) and extracted with ethyl acetate (3 X 100 mL). The combined organic phase was concentrated under reduced pressure and the residue was purified by silica-gel column chromatography eluting with petroleum ethyl acetate (6:1 to 3:1) to afford 195d (2.7 g, 96%). MS-ESI: [M-EtOH+H]+ 235. 1H NMR (500 MHz, DMSO) δ 7.35 (bs, 1H), 6.70 (bs, 1H), 6.60 (s, 1H), 4.60 (t, J =5.5 Hz, 1H), 4.18 (d, J =4.0 20 Hz, 2H), 3.57-3.56 (m, 2H), 3.25 (m, 2H), 2.57 (t, J = 6.0 Hz, 2H), 2.40 (t, J = 6.0 Hz , 2H), 1.71 (t, J =5.0 Hz, 2H), 1.64 (t, J =5.0 Hz, 2H), 1.01 (t, J = 7.0 Hz, 6H).
Example 195e 6,7,8,9-Tetrahydropyrazino[1,2-a]indol-1(2H)-one 195e A mixture of 195d (2.7 g, 9.6 mmol) and acetic acid (10 mL) was heated at 110oC for 2 h. The mixture was cooled to room ature and neutralized with aqueous sodium 25 carbonate on and extracted with ethyl acetate (3 X 30 mL). The ed organic phase was concentrated under reduced pressure to afford 195e as a yellow solid (1.6 g, 88%). MSESI : [M+H]+ 189.3. 1H NMR (500 MHz, DMSO-d6) δ 10.28 (s, 1H), 7.02 (d, J =5.5 Hz, 1H), 6.63 (s, 1H), , 6.52 (pt, J =5.5 Hz,1H), 2.66 (t, J = 6.0 Hz, 2H), 2.57 (t, J = 6.0 Hz , 2H), 1.83-1.82 (m, 2H), 1.73-1.72 (m, 2H). 30 Example 195f 4-Chloro(1-oxo-6,7,8,9-tetrahydropyrazino[1,2-a]indol- 2(1H)-yl)nicotinaldehyde 195f 267 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 1,4-dioxane (15 mL), 2-bromochloronicotinaldehyde 103a (503 mg, 2.28 mmol), 195e (142 mg, 0.76 mmol), cesium carbonate (490 mg, 1.5 5 mmol), CuI (143 mg, 0.76 mmol), and 4,7-dimethoxy-1,10-phenanthroline (127 mg, 0.52 mmol). After three cycles of vacuum/argon flush, the mixture was heated at 80oC for 10 hrs.
It was then cooled to room temperature and filtered. The filtrate was washed with brine and concentrated under reduced pressure. The resulting residue was purified with silica-gel column chromatography eluting with 1:4 ethyl acetate/petroleum ether to afford 195f (160 10 mg, 65%) as a yellow solid. : [M+H]+ 328.
Example 195g 2-(3-(Formyl)(1-methyl(5-methyl-4,5,6,7- tetrahydropyrazolo [1,5-a]pyrazinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)- 6,7,8,9-tetrahydro-pyrazino[1,2-a]indol-1(2H)-one 195g A 50-mL single-neck round-bottomed flask equipped with a reflux condenser was 15 charged with 195f (130 mg, 0.40 mmol), 1-methyl(5-methyl-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)(4,4,5,5-tetra-methyl-1,3,2-dioxaborolan yl)pyridin-2(1H)-one 135a (154 mg, 0.40 mmol), Pd(dppf)Cl2 (29 mg, 0.040 mmol), K3PO4 (170 mg, 0.80 mmol), sodium acetate (66 mg, 0.80 mmol), water (6 drops), and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at reflux for 2 h. It 20 was then cooled to room temperature and filtered. The filtrate was trated under reduced pressure and the resulting residue was purified by silica-gel column tography g with 30:1 dichloromethane/methanol to afford 195g as yellow solid (120 mg, 54%).
: [M+H]+ 551.2 Example 195 2-[3-(hydroxymethyl)[1-methyl[(5-methyl-6,7-dihydro-4H- 25 pyrazolo[1,5-a]pyrazinyl)amino]oxopyridyl]pyridyl]-6,7,8,9- tetrahydropyrazino[1,2-a]indolone 195 268 To a solution of 195g (120 mg, 0.22 mmol) in methanol (5 mL) at 0oC was added sodium borohydride (25 mg, 0.66 mmol). The reaction mixture was stirred for 30 minutes. It was then quenched with water (1.0 mL) and concentrated under reduced pressure. The residue was purified by reverse-phase PLC to afford 195 (70 mg, 58%). : 5 [M+H]+ 553.3. 1H NMR (500 MHz, CDCl3) δ 8.56 (d, J = 5.0 Hz, 1H), 7.99 (d, J = 2.0 Hz, 1H), 7.75 (d, J = 2.0 Hz, 1H), 7.49 (d, J = 4.5 Hz, 1H), 7.43 (s, 1H), 7.07 (s, 1H), 6.97 (d, J = 6.0 Hz, 1H), 6.67 (d, J = 6.0 Hz, 1H), 5.70 (s, 1H), 5.08-5.06 (m, 1H), 4.51-4.49 (m, 1H), 4.36-4.34 (m, 1H), 4.14-4.05 (m, 2H), 3.72 (s, 3H), 3.62-3.60 (m, 2H), .89 (m, 2H), 2.75-2.70 (m, 4H), 2.49 (s, 3H), 1.97-1.95 (m, 2H), 1.86-1.84 (m, 2H). 10 Example 196a 2-cyclopropylpyrimidinamine 196a Cyclopropylcarbamidine hydrochloride (1.0 g, 8.3 mmol) was ved in ethanol (25 mL) and triethylamine (1.26 g, 12.5 mmol), followed by the on of 2-chloroacrylonitrile (870 mg, 10 mmol). The resulting orange-yellow solution was refluxed for 1h. The 15 mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo and the residue was purified by reverse-phase Combiflash to afford 196a (300 mg, 27%) as a light brown solid. MS-ESI: [M+H]+ 136 Example 196b 5-Bromo(2-cyclopropylpyrimidinylamino) methylpyridin-2(1H)-one 196b 20 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 196a (300 mg, 2.22 mmol), 3,5-dibromomethylpyridin- 269 2(1H)-one (593 mg, 2.22 mmol), and cesium carbonate (1.45 g, 4.44 mmol). After ng nitrogen through the suspension for 30 s, Xantphos (127 mg, 0.22 mmol) and tris(dibenzyl-ideneacetone)dipalladium(0) (100 mg, 0.11 mmol) were added. The system was subject to three cycles of vacuum/argon flush and heated at reflux for 5 h. It was then cooled 5 to room temperature and filtered. The solid was washed with dichloromethane (2 X 50 mL).
The combined filtrate was concentrated under reduced pressure and the residue was washed with acetonitrile (5mL) to afford 196b (420 mg, 59%) as a yellow solid. MS-ESI: [M+H]+ 321 Example 196c 3-(2-Cyclopropylpyrimidinylamino)methyl(4,4,5,5- 10 tetramethyl-1,3,2- orolanyl)pyridin-2(1H)-one 196c A 100-mL single-neck bottomed flask equipped with a magnetic stirrer and a condenser was charged with 196b (380 mg, 1.2 mmol), Pin2B2 (1.5 g, 5.9 mmol), a)3 (55 mg, 0.060 mmol), X-phos (57 mg, 0.060 mmol), potassium acetate (350 mg, 3.6 mmol), and 1,4-dioxane (20 mL). The on mixture was subjected to three cycles of 15 vacuum/argon flush and was heated at 60°C for 15 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting e was washed with petroleum ether to afford 196c (410 mg, 94%) as yellow solid, which was used directly for next step without further purification. MS-ESI: [M+H]+ 369 Example 196d 4-{5-[(2-Cyclopropylpyridinyl)amino]methyloxo-1,6- 20 opyridinyl}{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- dienyl}pyridinecarbaldehyde 196d A 50-mL round-bottomed flask ed with a reflux ser was charged with 196c (258 mg, 0.70 mmol), 4-chloro{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 108a (240 mg, 25 0.70 mmol), K3PO4 (297 mg, 1.4 mmol), sodium acetate (190 mg, 1.4 mmol), 1,1’- bis(diphenylphosphino)ferrocenedichloropalladium(II) (29 mg, 0.035 mmol), acetonitrile (10 mL), and water (0.5 mL). After bubbling nitrogen through the mixture for 30 minutes, it was heated at 100°C under N2 protection for 1 h. Analysis of reaction by LCMS showed completed conversion to the desired product. The mixture was cooled to room temperature 30 and diluted with dichloromethane (20 mL) and water (20 mL). The aqueous layer was separated and extracted with dichloromethane (3 X 20 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The dark residue was purified by silica-gel column chromatography eluting with dichloromethane /methanol (80/1 to 30/1) to afford 196d (220 mg, 57%) as yellow solid. MS-ESI: [M+H]+ 549 270 Example 196 3-[4-[5-[(2-cyclopropylpyrimidinyl)amino]methyloxo pyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 196 To a solution of 196d (200 mg, 0.36 mmol) in methanol/dichloromethane(5/5 mL) 5 was added NaBH4 (42 mg, 1.1 mmol) at room temperature. After the reaction was stirred for 1 h, LCMS indicated the reaction was completed. The mixture was trated under reduced pressure and water (10 mL) was added to the residue. It was then extracted with dichloromethane (20 mL × 3). The combined organic layer was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced re. The residue was 10 purified by prep-HPLC to afford 196 (135 mg, 68%) as a white solid. MS-ESI: [M+H]+ 551. 1H NMR (500 MHz, CDCl 3) δ 8.84 (d, J = 2.5 Hz, 1H), 8.53 (d, J = 5.0 Hz, 1H), 8.22 (d, J = 6.0 Hz, 1H), 8.13 (d, J = 1.5 Hz, 1H), 8.01 (s, 1H), 7.38 (d, J = 5.0 Hz, 1H), 6.86 (s, 1H), 6.52 (d, J = 6.0 Hz, 1H), 5.22-5.19 (m, 1H), 4.72-4.69 (m, 1H), 4.56-4.54 (m, 1H), 4.31 (t, J = 11.0 Hz, 1H), 4.17 (d, J = 5.0 Hz, 2H), 3.94-3.91 (m, 1H), 3.74 (s, 3H), 2.58 (d, J = 5.5 Hz, 15 2H), 2.53 (s, 2H), 2.18-2.13 (m, 1H), 1.29 (s, 6H), 1.16-1.13 (m, 1H), 1.06-0.95 (m, 3H).
Example 197a tert-Butyl 4-(5-Nitropyridinyl)piperazinecarboxylate 197a A mixture of 2-bromonitropyridine (5.0 g, 24.6 mmol), tert-butyl piperazine ylate (13.8 g, 74.2 mmol), acetonitrile (150 mL) was stirred at reflux for 2.5 h. After 20 the reaction was completed, the solvent was d under reduced pressure to afford 197a as a yellow solid (4.1 g, 54%). MS-ESI: [M+H]+ 309. 271 Example 197b tert-Butyl 4-(5-Aminopyridinyl)piperazinecarboxylate 197b A 250-mL round-bottomed flask was purged with nitrogen and charged with 197a (4.0 g, 13.0 mmol), 10% palladium on carbon (10% wet, 500 mg), and methanol (130 mL). 5 The flask was evacuated, charged with hydrogen gas, and stirred at room temperature for 15 h. Hydrogen was then evacuated and nitrogen was charged to the flask. The catalyst was removed by filtration through a pad of CELITE® and the filtrate was concentrated under reduced pressure to afford 197b (3.3 g, 91%). MS-ESI: [M+H]+ 279 Example 197c tert-Butyl 4-(5-(6-Bromomethyloxo-3,4-dihydropyrazin- 10 2-ylamino)pyridinyl)piperazinecarboxylate 197c A mixture of 197b (500 mg, 1.8 mmol), 3,5-dibromomethylpyrazin-2(1H)-one (530 mg, 2.0 mmol), N-ethyl-N-isopropylpropanamine (1.5 mL, 0.90 mmol), and propan- 2-ol (20 mL) was stirred at 100oC for 15 h. After the on was completed, the solvent was removed under reduced pressure to afford 197c as a brown solid (375 mg, 45%). MS-ESI: 15 [M+H]+ 465.
Example 197d 5-Bromomethyl(6-(piperazinyl)pyridinylamino)- pyrazin-2(1H)-one 197d To a on of 197c (500 mg, 1.08 mmol) in dichloromethane (10 mL) was added 4.0 M HCl/dioxane (10 mL). The reaction mixture was stirred at room temperature for 5 h. It 20 was then concentrated under reduced pressure to afford 197d (358 mg, 91%). : [M+H]+ 365.
Example 197e omethyl(6-(4-(oxetanyl)piperazinyl)pyridin- 3-ylamino)pyrazin-2(1H)-one 197e A mixture of 197d (0.75 g, 2.1 mmol), oxetanone (0.24 mL, 4.2 mmol), N 25 (0.32 g, 5.1 mmol), and zinc chloride/diethyl ether (5.1 mL , 5.1 mmol) in methanol (30 mL) was d at 50oC for 5 hours. The solid was removed by filtration and the filtrate was concentrated under reduced pressure. The e was purified by silica-gel column chromatography eluting with 10:1 romethane/methanol to afford 197e (550 mg, 64%).
MS-ESI: [M+H]+ 421. 30 Example 197f (4-(4-Methyl(6-(4-(oxetanyl)piperazinyl)pyridin ylamino)oxo- 4,5-dihydropyrazinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)pyridinyl)methyl Acetate 197f 272 A round-bottomed flask equipped with a reflux condenser was charged with 197e (200 mg, 0.48 mmol), 3-(acetoxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)pyridinylboronic acid 113i (364 mg, 0.95 mmol), PdCl2(dppf) (40 mg, 5 0.049 mmol), K3PO4 3H2O ( 250 mg, 0.95 mmol), sodium acetate(80 mg, 0.95 mmol), itrile (10 mL), and water (0.5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 80oC for 3 h. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified by silica-gel column chromatography eluting with 1:20 methanol/dichloromethane to afford 197f as a red solid (230 mg, 70%). MS-ESI: [M+H]+ 680 10 Example 197 hydroxymethyl)[4-methyl[[6-[4-(oxetan yl)piperazinyl]pyridyl]amino]oxo-pyrazinyl]pyridyl]-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indolone 197 A mixture of 197f (200 mg, 0.30 mmol) and lithium hydroxide (70 mg, 3.0 mmol) in THF (9 mL), i-propanol (6 mL), and water (1 mL) was stirred at room temperature for 0.5 h. 15 The mixture was trated under reduced pressure and diluted with water (4 mL). It was then extracted with dichloromethane (2 X 10 mL) and the combined dichloromethane extract was concentrated under reduced pressure. The e was purified with reverse-phase prep- HPLC to afford 197 (59 mg, 30%) as yellow solid. MS-ESI: [M+H]+ 638. 1H NMR (500 MHz, DMSO-d6) δ 9.30 (s, 1 H), 8.70 (d, J = 2.5 Hz, 1 H), 8.49 (d, J = 5.5 Hz, 1 H), 8.14- 20 8.11 (m, 1 H), 7.59 (s, 1 H), 7.55 (d, J = 5.0 Hz, 1 H), 6.82 (d, J = 9.5 Hz, 1 H), 6.58 (s, 1 H), 4.93 (t, J = 5.5 Hz, 1 H), 4.60-4.54 (m, 3 H), 4.48-4.42 (m, 3 H), .08 (m, 3 H), 3.86 (d, J = 12.0 Hz, 1 H), 3.54 (s, 3 H), 3.44-3.40 (m, p, 5 H), 2.66-2.53 (m, 2 H), 2.46-2.47 (m, 2 H), 2.35-2.33 (m, 4 H), 1.80-1.68 (m, 4 H).
Example 198a 4-(1-Methyl(5-(4-(oxetanyl)piperazinyl)pyridin 25 ylamino)oxo-1,6-dihydropyridinyl)(4-oxo-7,8,9,10-tetrahydropyridazino[4,5- a]indolizin-3(4H)-yl)nicotinaldehyde 198a 273 A 50-mL round bottomed flask equipped with a reflux condenser was charged with 4- chloro(4-oxo-7,8,9,10-tetrahydropyridazino[4,5-a]indolizin-3(4H)-yl)nicotinaldehyde 192c (118 mg, 0.36 mmol), 1-methyl(5-(4-(oxetanyl)piperazinyl)pyridinyl- 5 amino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 101l (171 mg, 0.36 mmol), Na2CO3 (78 mg, 0.72 mmol), Pd (dppf)Cl2 (30 mg, 0.036 mmol), DMF (10 mL), and water (1 mL). After bubbling nitrogen through the e for 30 minutes, it was heated at 50 oC for 10 hours under N 2 protection. It was then cooled to room temperature and evaporated under reduced pressure. The residue was purified by silica-gel column chromatography 10 eluting with 20:1 dichloromethane/methanol to afford 198a (93 mg, 40%) as a brown solid.
MS-ESI: [M+H]+ 634.3.
Example 198 3-[3-(hydroxymethyl)[1-methyl[[5-[4-(oxetanyl)piperazin yl]pyridyl]amino]oxopyridyl]pyridyl]-7,8,9,10-tetrahydropyridazino[4,5- lizinone 198 15 To a solution of 198a (80 mg, 0.13 mmol) in methanol (4 mL) was added NaBH4 (14 mg, 0.39 mmol). The mixture was stirred at 20oC for 2 h. It was then evaporated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 198 (38 mg, 43%) as an off-white solid.MS-ESI: [M+H]+ 636.4. 1H NMR (500 MHz, DMSO-d6) δ 8.65 (d, J = 2.0 Hz, 1H), 8.54 (d, J = 5.0 Hz, 1H), 8.44 (s, 1H), 8.25 (s, 1H), 7.88 (d, J = 2.5 20 Hz, 1H), 7.62 (s, 1H), 7.50 (d, J = 2.0 Hz, 1H), 7.46 (d, J = 4.5 Hz, 1H), .36 (m, 1H), 7.25-7.23 (m, 1H), 4.67 (bs, 1H), 4.55 (t, J = 6.5 Hz, 2H), 4.46 (t, J = 6.0 Hz, 2H), 4.33-4.31 (m, 2H), 4.26-4.20 (m, 2H), 3.59 (s, 3H), 3.46-3.41 (m, 1H), 3.09-3.03 (m, 6H), 2.39-2.37 (m, 4H), 2.04-1.96 (m, 2H), 1.93-1.86 (m, 2H).
Example 199a Imidazo[1,2-a]pyrimidinamine 199a 274 Br N O O 2 eq Cl N N NH N H N N Br O H2N N NH2 N EtOH/H2O, N NH2 Pd2(dba)3, Xanphos N Br NaHCO3 Cs2CO3, dioxane 70°C 199a 199b To the solution of pyrimidine-2,4-diamine (3.0 g, 0.027 mol) in ethanol (90 mL) and aqueous NaHCO3 (2M, 20 mL) was added 2-chloroacetaldehyde (4.3 g, 0.055 mol). The mixture was stirred at 70oC overnight. TLC showed the starting material disappeared. The 5 solvent was removed under d pressure and the residue was extracted with ethyl acetate (3 X 30 mL). The combined organic layer was concentrated under reduced pressure and the residue was purified by silica-gel column chromatography eluting with 1:5 petroleum ethyl acetate to afford 199a as a white solid (2.2 g, 60%). MS: [M+H]+ 135.1. e 199b 5-Bromo(imidazo[1,2-a]pyrimidinylamino) 10 methylpyridin-2(1H)-one 199b A 250-mL round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 199a (2.2 g, 16.4 mmol), 3,5-dibromomethylpyridin-2(1H)- one (8.77 g, 32.8 mmol), Pd2dba3 (1.5 g, 1.64 mmol), os (1.88 g, 3.28 mmol), Cs2CO3 (10.7 g. 32.8 mmol), and 1,4-dioxane (150 mL). The system was evacuated and then ed 15 with N2. It was then heated at reflux for 3 h. After the completion of the reaction, the mixture was filtered off and the solid was washed with methanol (60 mL). The combined filtrate was evaporated under reduced pressure and the residue was purified by silica-gel column chromatography eluting with 20:1 dichloromethane /methanol to afford 199b as a light green solid (1.63 g, 31%). MS: [M+H]+ 320.1 20 Example 199c chloro(hydroxymethyl)pyridinyl]-4,4-dimethyl- iazatricyclo [6.4.0.02,6]dodeca-2(6),7-dienone 199c 275 O AcCl, Et3N N NaBH4 (5.0 eq) OH OAc N THF, rt, 1 h N N Cl DCM/CH3OH, 0°C, 1h N Cl N Cl O N O N O N 108a 199c 199d N O O B B 199b N N NH O O AcO N OH AcO O N N B OH N N Pd(dppf)Cl2 Pd(dppf)Cl2 (0.05 eq) x-phos O N K3PO4 (2 eq) O N KOAc NaOAc 3H2O(2 eq) dioxane, 65 oC, 4 h 199f 199e H2O, CH3CN, 100 oC, 2h A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with 4-chloro{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- 0-yl}pyridinecarbaldehyde 108a (9.0 g, 26.1 mmol, 1.0 eq.), methanol (50 mL), 5 romethane (30 mL), and NaBH4 (5.95 g, 156.6 mmol, 5.0 eq.) at 0OC. The reaction mixture was stirred for 1 h. After the reaction was completed, the reaction was quenched with water and concentrated under d pressure. The residue was extracted with dichloromethane. The combined organic layer was dried over Na2SO4 and concentrated under reduced pressure. The residue was purified by silica-gel column tography eluting with 10 1:4 ethyl acetate/petroleum ether to afford 199c as a white solid (7.0 g, 77%). MS-ESI: [M+H]+ 345.9.
Example 199d (4-Chloro{4,4-dimethyloxo-1,10-diazatricyclo- [6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl)methyl Acetate 199d A 250-mL -neck round-bottomed flask equipped with a ic stirrer was 15 charged with 199c (7.0 g, 20.2 mmol, 1.0 eq.), triethylamine (4.08 g, 40.4 mmol, 2.0 eq.), and THF (50 mL). To the mixture was added the solution of acetyl chloride (2.36 g, 30.3 mmol, 1.5 eq.) in THF (20 mL) dropwise. The reaction mixture was stirred at room temperature for one hour. After the reaction was completed, it was quenched with ice water and evaporated under reduced pressure. The residue was extracted with dichloromethane. The combined 20 organic layer was dried over Na2SO4 and concentrated under reduced pressure. The resulting residue was washed with 1:8 ethyl acetate/petroleum to afford 199d as a white solid (5.9 g, 76%). MS-ESI: [M+H]+ 388.3.
Example 199e {3-[(Acetyloxy)methyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6] dodeca-2(6),7-dienyl}pyridinyl}boronic Acid 199e 276 A 250-mL single-neck round-bottomed flask ed with a magnetic stirrer and a reflux condenser was charged with 199d (4.5 g, 1.0 eq., 11.6 mmol), Pin2B2 (7.38 g, 2.5 eq., 29.0 mmol), PdCl2(dppf) (473 mg, 0.05 eq., 0.58 mmol), x-phos (470 mg, 0.1 eq., 1.16 mmol), potassium acetate (3.41 g, 3.0 eq., 34.8 mmol), and dioxane (100 mL). After three 5 cycles of vacuum/argon flush, the mixture was heated at 65ºC for 4 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure to afford crude 199e as a brown-red liquid (4.0 g, purity: 65%). MS-ESI: [M+H]+ 398.3.
Example 199f (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}[5-({imidazo[1,2-a]pyrimidinyl}amino)methyloxo-1,6- 10 dihydropyridinyl]pyridin yl)methyl Acetate 199f A 100-mL single-neck round-bottomed flask equipped with a reflux condenser was charged with 199b (500 mg, 1.5 mmol), {3-[(acetyloxy)methyl]{4,4-dimethyloxo-1,10- diazatri-cyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (1200 mg, 3.0 mmol), Pd(dppf)Cl2 (65 mg, 0.075 mmol), K3PO4 (650 mg, 3.0 mmol), sodium acetate 15 trihydrate (420 mg, 3.0 mmol), water (6 drops), and acetonitrile (20 mL). The system was evacuated and refilled with N2. The reaction mixture was heated at 100ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was ed by silica-gel column chromatography eluting with 25:1 dichloromethane/methanol to afford 199f (240 mg, 40%) as a yellow-brown solid. MS-ESI: 20 [M+H]+ 593.4. e 199 3-[3-(hydroxymethyl)[5-(imidazo[1,2-a]pyrimidinylamino) methyloxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 199 A mixture of 199f (180 mg, 0.30 mmol) and lithium hydroxide (130 mg, 3.0 mmol) in 25 i-propanol/THF (5:3, 8 mL) and water (2 mL) was stirred at 30oC for 1 h. The mixture was evaporated in vacuo and the residue was d with water (3 mL). It was then ted with ethyl acetate (2 X 10 mL). The combined ethyl acetate extract was concentrated under reduced pressure and the residue was purified by prep-HPLC to afford 199 (40 mg, 30 %) as white solid. MS-ESI: [M+H]+ 551.3. 1H NMR (500 MHz, CHCl3) δ 9.09 (d, J = 1.5 Hz, 1H), 30 8.51 (d, J = 5.0 Hz, 1H), 8.23 (s, 1H), 8.13 (d, J = 7.0 Hz, 1H), 7.93 (d, J = 2.0 Hz, 1H), 7.45 (s, 1H), 7.38 (d, J = 5.0 Hz, 1H), 7.27 (s, 1H), 6.84 (s, 1H), 6.47 (d, J = 7.5 Hz, 1H), 5.10 (s, 1H), 4.67-4.50 (m, 2H), 4.32-4.18 (m, 3H), 3.93-3.88 (m, 1H), 3.74 (s, 3H), 2.60-2.58 (m, 2H), 2.52 (s, 2H), 1.28 (s, 6H). 277 Example 200a tert-Butyl 4-(6-Nitropyridinyl)-5,6-dihydropyridine-1(2H)- carboxylate 200a Br Boc O N Boc Pd/C, H2, MeOH N N 20 °C, 12 h Br N Pd2(dba)3, xantphos,Cs2CO3 NO2 78% N NH2 Dioxane, 100 °C, 6 h 61 % 200a 200b Boc HN N O N NH HCl / Dioxane N NH O 25 °C, 4 h O O ZnCl2, NaCNBH3, 84 % N N Br MeOH, 50 °C, 5 h Br 78% 200d 200c O O N N N NH N NH AcO O 113i O N N N N 0.1eq PdCl2(dppf), 2eq K3PO4 Br 2eq CH3COONa, H2O, CH3CN, 3h O N 200e 200f A mixture of 5-bromonitropyridine (2.0 g, 9.7 mmol), tert-butyl 4-(4,4,5,5- 5 tetramethyl-1,3,2-dioxaborolanyl)-5,6-dihydropyridine-1(2H)-carboxylate (3.0 g, 9.7 mmol), Pd(dppf)Cl2 (792 mg, 0.97 mmol), K3PO4.3H2O (5.2 g, 19.4 mmol), and sodium acetate (1.59 g, 19.4 mmol) in acetonitrile (100 mL) and water (5 mL) was evacuated and then refilled with N2. The reaction mixture was heated at 80ºC for 6 h. It was then cooled to room temperature and filtered. The te was trated under reduced pressure and the 10 resulting residue was purified by silica-gel column chromatography eluting with 1:5 ethyl acetate/petroleum ether to afford 200a as a yellow solid (2.2 g, 74%). e 200b tert-Butyl 4-(6-Aminopyridinyl)piperidinecarboxylate 200b 278 A 500-mL round-bottomed flask was purged with nitrogen and charged with 200a (2.5 g, 8.2 mmol), 10% palladium on carbon (50% wet, 300 mg), and methanol (80 mL). The flask was evacuated, charged with en gas, and d at room temperature under hydrogen atmosphere for 12 h. The hydrogen was then evacuated and nitrogen was charged 5 to the flask. The catalyst was d by filtration through a pad of CELITE® and the filtrate was concentrated under reduced pressure to afford 200b (1.8 g, 78%) as a white solid.
MS-ESI: [M+H]+ 278.1 Example 200c tert-Butyl 4-(6-(5-Bromomethyloxo-1,2-dihydropyridin- 3-ylamino)pyridinyl)-5,6-dihydropyridine-1(2H)-carboxylate 200c 10 A 100-mL single-neck round-bottomed flask ed with a magnetic stirrer and a reflux condenser was charged 200b (2.0 g, 7.2 mmol), 3,5-dibromomethylpyridin-2(1H)- one (1.9 g, 7.2 mmol), cesium ate (4.7 g, 14.4 mmol), and 1,4-dioxane (50 mL). After bubbling nitrogen through the resulting mixture for 30 min, Xantphos (418 mg, 0.72 mmol) and tris(dibenzylideneacetone)dipalladium(0) (661 mg, 0.72 mmol) were added. The reaction 15 mixture was subject to three cycles of vacuum/argon flush and heated at 100oC for 6 h. After this time the reaction was cooled to room ature and filtered. The filtrate was partitioned between ethyl acetate (120 mL) and water (60 mL). The aqueous layer was separated and extracted with ethyl e (3 × 80 mL). The combined organic layer was washed with brine (30 mL) and dried over sodium sulfate. The drying agent was removed by 20 filtration and the filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column tography eluting with 1:4 ethyl e/petroleum ether to afford 200c (2.0 g, 61%) as a yellow solid. MS-ESI: [M+H]+ 463.2 Example 200d 5-Bromomethyl(5-(1,2,3,6-tetrahydropyridin yl)pyridinylamino)pyridin-2(1H)-one 200d 25 A mixture of 200c (1.0 g, 2.3 mmol) and 4 M HCl/dioxane (10 mL) was stirred at room temperature for 4 h. The mixture was concentrated under reduced pressure. The residue was basified with aqueous sodium hydroxide and extracted with dichloromethane. The combined organic layer was washed with water and brine, dried over Na2SO4, and concentrated under reduced pressure to afford 200d (650 mg, 84%) as a yellow solid. MS- 30 ESI: [M+H]+ 363.0 Example 200e 5-Bromomethyl(5-(1-(oxetanyl)-1,2,3,6- tetrahydropyridinyl)pyridinylamino)pyridin-2(1H)-one 200e A mixture of 200d (500 mg, 1.4 mmol), oxetanone (298 mg, 4.2 mmol), NaBH3CN (261 mg, 4.2 mmol), and 1 mol/L zinc chloride in ethoxyethane (4 mL, 4.2 mmol) in 279 methanol (20 mL) was stirred at 50oC for 5 hours. Water (20 mL) was added to the reaction and the resulting e was extracted with dichloromethane (3 × 50 mL). The combined organic layer was concentrated under reduced re and the residue was purified by silicagel column chromatography eluting with 10:1 methylene chloride/methanol to afford 200e 5 (450 mg, 78%) as a yellow solid. MS-ESI: [M+H]+ 419.1 Example 200f (4-(1-Methyl(5-(1-(oxetanyl)-1,2,3,6-tetrahydropyridin yl)pyridinylamino)oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl Acetate 200f A round-bottomed flask equipped with a reflux condenser was charged with 200e 10 (300 mg, 0.72 mmol), 3-(acetoxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)pyridinylboronic acid 113i (414 mg, 1.08 mmol), PdCl2(dppf) (57 mg, 0.070 mmol), K3PO4 3H2O (560 mg, 2.16 mmol), sodium acetate (177 mg, 2.16 mmol), acetonitrile (10 mL), and water (0.5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 80oC for 3 h. It was then filtered and the filtrate was evaporated in 15 vacuo. The e was purified with silica-gel column chromatography eluting with 1:20 methanol/dichloromethane to afford 200f as a red solid (324 mg, 68%). MS-ESI: [M+H]+ 676.2 Example 200 2-[3-(hydroxymethyl)[1-methyl[[5-[1-(oxetanyl)-3,6-dihydro- 2H-pyridinyl]pyridyl]amino]oxopyridyl]pyridyl]-3,4,6,7,8,9- 20 hexahydropyrazino[1,2-a]indolone 200 A e of 200f (260 mg, 0.39 mmol) and lithium hydroxide (92.4 mg, 3.85 mmol) in THF (9 mL), isopropanol (6 mL), and water (1 mL) was stirred at room temperature for 0.5 h. The e was ted concentrated under reduced pressure and diluted with water (4 mL). It was then extracted with romethane (2 X 10 mL) and the combined C 25 dichloromethane extract was concentrated under reduced pressure. The residue was ed with reverse-phase prep-HPLC to afford 200 (53.1 mg, 20%) as yellow solid. MS-ESI: [M+H]+ 634.2. 1H NMR (500 MHz, DMSO-d6) δ 8.76 (d, J = 1.5 Hz, 1 H), 8.74 (s, 1H), 8.49 (d, J = 5.5 Hz, 1 H), 8.24 (d, J = 2.0 Hz, 1 H), 7.73-7.71 (m, 1 H), 7.54 (d, J = 2.5 Hz, 1 H), 7.36 (d, J = 5.0 Hz, 1 H), 7.30 (d, J = 8.5 Hz, 1 H), 6.58 (s, 1 H), 6.11 (s, 1 H), 4.97 (s, 1 H), 30 4.57 (t, J = 6.5 Hz, 2 H), 4.38-4.49 (m, 4 H), 4.08-4.26 (m, 3 H), 3.86 (d, J = 12.0 Hz, 1 H), 3.61 (s, 3 H), 3.54-3.45 (m, 1 H), 2.95 (s, 2 H), 2.68-2.54 (m, 2 H), 2.48-2.46 (m, overlap, 6 H),1.83-1.75 (m, 2 H), 1.73-1.65 (m, 2 H).
Example 201a 5-Bromo(imidazo[1,2-a]pyridinylamino) methylpyrazin-2(1H)-one 201a 280 A 100-mL round-bottomed flask equipped with a reflux condenser was charged with imidazo[1,2-a]pyridinamine (665 mg, 5.0 mmol), Cs2CO3 (3.26 g, 10 mmol), 3,5- dibromomethylpyrazin-2(1H)-one (1.86 g, 7.0 mmol), Xantphos (289 mg, 0.50 mmol), 5 Pd2(dba)3 (458 mg, 0.50 mmol), and 1, 4-dioxane (30 mL). After bubbling nitrogen through the mixture for 30 minutes, it was heated at 100°C under nitrogen atmosphere for 16 h.
Analysis of the reaction mixture by LCMS showed little ng material remained. The reaction mixture was cooled to room temperature and filtered. The te was diluted with dichloromethane (60 mL) and water (50 mL). The aqueous layer was separated and extracted 10 with dichloromethane (3 × 20 mL). The combined organic layers was dried over Na2SO4, filtered, and concentrated under reduced re. The dark residue was purified by -gel column chromatography eluting with dichloromethane /methanol (60/1 to 30/1) to afford 201a (700 mg, 44%) as light yellow solid. MS-ESI: [M+H]+ 320 Example 201b (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 15 2(6),7-dienyl}[6-({imidazo[1,2-a]pyridinyl}amino)methyloxo-4,5- dihydropyrazinyl]pyridinyl)methyl Acetate 201b A 25-mL single-neck round-bottomed flask ed with a reflux condenser was charged with 201a (64 mg, 0.20 mmol), {3-[(acetyloxy)methyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (160 mg, 20 0.40 mmol), Pd(dppf)Cl2 (10 mg, 0.012 mmol), K3PO4 (100 mg, 0.39 mmol), NaOAc·3H2O (60 mg, 0.44 mmol), water (6 drops), and acetonitrile (5 mL). The system was evacuated and refilled with N2. The reaction e was d at 100ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under d pressure and the resulting residue was purified by silica-gel column chromatography eluting with 20:1 25 dichloromethane/methanol to afford 201b (40 mg, 34%) as a yellow brown solid. MS-ESI: [M+H]+ 593.2. 281 Example 201 3-[3-(hydroxymethyl)[6-(imidazo[1,2-a]pyridinylamino) methyloxo-pyrazinyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 201 A mixture of 201b (40 mg, 0.067 mmol) and lithium hydroxide (25 mg, 0.60 mmol) 5 in i-propanol/THF (3:2, 5 mL) and water (1 mL) was stirred at 30oC for 1 h. The mixture was evaporated in vacuo and the residue was extracted with ethyl e (2 X 10 mL). The combined ethyl acetate extract was concentrated under reduced pressure and the residue was purified by reverse-phase PLC to afford 201 (10 mg, 30%) as a white solid. MS-ESI: [M+H]+ 551.3. 1H NMR (500 MHz, CHCl3) δ 9.58 (s, 1H), 8.58 (d, J = 5.0 Hz, 1H), 8.18 (s, 10 1H), 8.12 (s, 1H), 7.73 (d, J = 5.0 Hz, 1H), 7.63-7.61 (m, 2H), 7.55 (s, 1H), 7.13-7.11 (m, 1H), 6.87 (s, 1H), 5.19-5.17 (m, 1H), .75 (m, 1H), 4.57-4.42 (m, 2H), 4.20-4.17 (m, 2H), 3.92-3.90 (m, 1H), 3.70 (s, 3H), 2.60-2.53 (m, 4H), 1.29 (s, 6H).
Example 202a ethyl(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinylamino)oxo-1,6-dihydropyridinyl)(4-oxo-7,8,9,10- 15 tetrahydropyridazino[4,5-a]indolizin-3(4H)-yl)nicotinaldehyde 202a A 100-mL round-botommed flask equipped with a reflux condenser was charged with 4-chloro(4-oxo-7,8,9,10-tetrahydropyridazino[4,5-a]indolizin-3(4H)-yl)nicotinaldehyde 192c (200 mg, 0.60 mmol), 1-methyl(5-methyl-4,5,6,7-tetrahydropyra-zolo[1,5-a]pyrazin- 20 2-ylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 135a (230 mg, 0.60 mmol), sodium acetate (100 mg, 1.2 mmol), K3PO4 (320 mg, 1.2 mmol), PdCl2(dppf) (50 mg, 0.060 mmol), acetonitrile (25 mL), and water (1 mL). After bubbling nitrogen through the mixture for 30 minutes, it was heated at 100oC for 3 hours under N2 protection.
The on was cooled to room temperature and evaporated under reduced pressure. The 25 residue was purified by silica-gel column chromatography eluting with 25:1 ene chloride/methanol to afford 202a (205 mg, 62%) as a brown solid. MS-ESI: [M+H]+ 552.3. 282 Example 202 3-[3-(hydroxymethyl)[1-methyl[(5-methyl-6,7-dihydro-4H- pyrazolo[1,5-a]pyrazinyl)amino]oxopyridyl]pyridyl]-7,8,9,10- tetrahydropyridazino[4,5-a]indolizinone 202 To a solution of 202a (180 mg, 0.33 mmol) in methanol (25 mL) was added NaBH4 5 (37 mg, 0.99 mmol). The mixture was stirred at 20oC for 2 h and quenched with water. It was then evaporated under reduced pressure and the residue was ed by reverse-phase prep- HPLC to afford 202 (120 mg, 66%) as a white solid. MS-ESI: [M+H]+ 554.3. 1H NMR (500 MHz, DMSO-d6) δ 8.53 (d, J = 5.0 Hz, 1H), 8.25 (s, 1H), 8.19 (s, 1H), 8.07 (d, J = 2.0 Hz, 1H), 7.61 (s, 1H), 7.45 (d, J = 5.0 Hz, 1H), 7.43 (d, J = 2.5 Hz, 1H), 5.89 (s, 1H), 4.65 (t, J = 10 5.0 Hz, 1H), 4.34-4.32 (m, 2H), 4.26-4.20 (m, 2H), 3.93-3.91 (m, 2H), 3.58 (s, 3H), 3.49 (s, 2H), 3.06-3.04 (m, 2H), 2.79-2.77 (m, 2H), 2.35 (s, 3H), 2.04-1.96 (m, 2H), 1.93-1.86 (m, 2H).
Example 203a 5-Methyl-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridinamine 203a Br O O N N Br N NH2 NH2 N S N S, pyrrolidine Pd2(dba)3, Xantphos, iPA, 5h Cs2CO3, dioxane, 203a 100 °C, 15 h N N S S N NH N NH O O O 199e N N N N Br Pd(dppf)Cl2, K3PO4, O N NaOAc, CH3CN, H2O 203b 100 °C, 1h 15 203c A solution of 1-methylpiperidone (11.3 g, 100 mmol) in anol (80 mL) was heated to 50ºC. To the on were sequentially added a solution of cyanamide (4.2 g, 100 mmol) in 2-propanol (25 mL) and sulfur powder (3.2 g, 100 mmol). After a tic amount of pyrrolidine (1.3 mL) was added, the resultant mixture was stirred at 50 ºC for 2 hours. The 20 reaction mixture was allowed to cool to room temperature and stirred overnight. It was then cooled to or below 10 ºC in an ice-water bath and stirred for 1 hour at the same temperature.
The precipitated crystals were collected by filtration and washed with 2-propanol (20 mL). 283 The wet ls were dried in vacuum to afford 203a (10 g, 59%). MS: [M+H]+ 170. 1H NMR (500 MHz, DMSO-d6) δ 6.70 (s, 2H), 3.31 (s, 2H), 2.61 (t, J = 5.5 Hz, 2H), 2.45 (m, 2H), 2.33 (s, 3H).
Example 203b 5-Bromomethyl(5-methyl-4,5,6,7-tetrahydrothiazolo[5,4- 5 c]pyridinylamino)pyridin-2(1H)-one 203b Following the procedures described for 191g and starting with 203a (4.0 g, 23.5 mmol) and 3,5-dibromomethylpyridin-2(1H)-one (3.0 g, 17.8 mmol) afforded 203b as yellow solid (2.8 g, 44%). MS: [M+H]+ 357.
Example 203c 10-[3-(Acetoxymethyl)[1-methyl({5-methyl-4H,5H,6H,7H- 10 [1,3]thiazolo[5,4-c]pyridinyl}amino)oxo-1,6-dihydropyridinyl]pyridinyl]-4,4- dimethyl-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienone 203c A 50-mL round-bottomed flask equipped with a reflux condenser was charged with 203b (178 mg, 0.50 mmol), {3-[(acetyloxy)methyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (200 mg, 15 0.50 mmol), K3PO4 (212 mg, 1.0 mmol), sodium acetate (82 mg, 1.0 mmol), 1,1’- bis(diphenylphosphino)ferrocene-dichloropalladium(II) (21 mg, 0.025 mmol), acetonitrile (10 mL), and water (0.5 mL). After bubbling nitrogen through the mixture for 30 minutes, it was heated at 100°C under N2 protection for 1h. Analysis of reaction mixture by LCMS showed complete conversion to the desired t. The reaction mixture was cooled to room 20 ature and filtered. The filtrate was concentrated under reduced pressure and the residue was diluted with dichloromethane (20 mL) and water (10 mL). The aqueous layer was separated and extracted with dichloromethane (2 × 20 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under d pressure. The dark residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (80/1 25 to 30/1) to afford 203c (135 mg, 43%) as yellow solid. MS-ESI: [M+H]+ 584 e 203 3-[3-(hydroxymethyl)[1-methyl[(5-methyl-6,7-dihydro-4H- thiazolo[5,4-c]pyridinyl)amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 203 To a on of 203c (140 g, 0.22 mmol) in THF/i-propanol/water(5/2/2 mL) was 30 added LiOH (54 mg, 2.2 mmol) at room temperature. After the on was d for 1 h, LCMS indicated the reaction was completed. Then the mixture was concentrated under reduced pressure and diluted with water (3 mL). It was then extracted with dichloromethane (3 X 10 mL). The combined organic layer was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep- 284 HPLC to afford 203 (85 mg, 66%) as white solid. MS-ESI: [M+H]+ 586. 1H NMR (500 MHz, CDCl3) δ 8.50 (d, J = 5.5 Hz, 1H), 8.37 (d, J = 2.0 Hz, 1H), 8.32 (s, 1H), 7.95 (d, J = 2.0 Hz, 1H), 7.34 (d, J = 5.0 Hz, 1H), 6.85 (s, 1H), 5.11-5.09 (m, 1H), 4.67-4.64 (m, 1H), 4.52 (bs, 1H), 4.30-4.28 (m, 1H), 4.16 (d, J = 4.5 Hz, 2H), 3.89-3.86 (m, 1H), 3.72 (s, 3H), 3.60 (s, 5 2H), 2.84-2.81 (m, 4H), 2.58 (d, J = 5.0 Hz, 2H), 2.53 (s, 3H), 2.52 (s, 2H), 1.28 (s, 6H).
Example 204a 4-(1-Methyl(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinylamino)oxo-1,6-dihydropyridinyl)(1-oxo-6,7,8,9- tetrahydropyridazino[4,5-b]indolizin-2(1H)-yl)nicotinaldehyde 204a 10 A 100-mL round-bottomed flask equipped with a reflux condenser was charged with 4-chloro(1-oxo-6,7,8,9-tetrahydropyridazino[4,5-b]indolizin-2(1H)-yl)nicotinaldehyde 193f (200 mg, 0.60 mmol), 1-methyl(5-methyl-4,5,6,7-tetrahydropyrazolo-[1,5-a]pyrazin- 2-ylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 135a (230 mg, 0.60 mmol), sodium acetate (100 mg, 1.2 mmol), K3PO4 (320 mg, 1.2 mmol), PdCl2(dppf) 15 (50 mg, 0.060 mmol), acetonitrile (25 mL), and water (1 mL). After bubbling en through the resulting e for 30 minutes, the e was heated at 100oC for 3 hours.
The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 20:1 methylene chloride/methanol to afford 204a (185 mg, 55%) as a brown solid. MS-ESI: 20 [M+H] + 552.3. e 204 hydroxymethyl)[1-methyl[(5-methyl-6,7-dihydro-4H- pyrazolo[1,5-a]pyrazinyl)amino]oxopyridyl]pyridyl]-6,7,8,9- tetrahydropyridazino[4,5-b]indolizinone 204 To a solution of 204a (160 mg, 0.29 mmol) in methanol (20 mL) was added NaBH4 25 (33.0 mg, 0.87 mmol). The mixture was stirred at 20oC for 2 h and quenched with water. It was then evaporated under reduced pressure and the residue was purified by reverse-phase 285 prep-HPLC to afford 204 (120 mg, 75%) as a white solid. MS-ESI: [M+H] + 554.3. 1H NMR (500 MHz, DMSO-d6) δ 8.55 (d, J = 5.0 Hz, 1H), 8.46 (s, 1H), 8.23 (s, 1H), 8.09 (d, J = 2.5 Hz, 1H), 7.49 (d, J = 5.0 Hz, 1H), 7.40 (d, J = 2.0 Hz, 1H), 6.49 (s, 1H), 5.89 (s, 1H), 4.73 (bs, 1H), 4.30 (s, 2H), 4.27-4.25 (m, 2H), 3.93-3.91 (m, 2H), 3.58 (s, 3H), 3.49 (s, 2H), 2.95-2.93 5 (m, 2H), 2.78-2.76 (m, 2H), 2.34 (s, 3H), 2.04-1.99 (m, 2H), 1.88-1.83 (m, 2H).
Example 205a 2-Nitro-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine 205a Br Boc Boc O N Boc2O, Boc HN cat.DMAP N Pd/C, MeOH N N THF, r.t. 2 h, rt Br N N N NH N N N NO2 O N NO2 N NH2 a)3, Xantphps, N 205b 205a 205c Cs2CO3, e, Br 100 oC, 4 h 205d Boc OAc N OH N HN N B OH N N N NH O N N NH HCl/dioxane, OAc O OAc O N CH2Cl2 N 113i rt, 2 h N N N N O N f)Cl2, K3PO4, NaOAc, O N CH3CN, H2O, 2 h, 100 oC 205e 205f A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with 1-(2-bromoethyl)(bromomethyl)nitro-1H-pyrazole 125c (3.0 g, 9.64 10 mmol) in THF (35 mL) and aqueous a (135 mL, ). The mixture was d at room temperature for 72 h under nitrogen. The reaction mixture was then concentrated under reduced pressure and the resulting residue was partitioned n ethyl acetate (100 mL) and water (100 mL). The aqueous layer was extracted with ethyl acetate (2 × 50 mL). The combined organic layer was washed with 10% potassium carbonate (2 × 100 mL), brine (200 15 mL), and dried over sodium sulfate. The drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to afford 205a as a yellow solid (1.23 g, 76%). MS: [M+H]+ 169 Example 205b tert-Butyl 2-Nitro-6,7-dihydropyrazolo[1,5-a]pyrazine-5(4H)- carboxylate 205b 20 To a solution of 205a (504 mg, 3.0 mmol) in THF (20 mL) was added (Boc)2O (785 mg, 3.60 mmol) and DMAP (74 mg, 0.60 mmol). The reaction mixture was stirred at room temperature overnight. Then it was filtered and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica-gel column chromatography eluting 286 with 100:1 dichloromethane/methanol to afford 205b as white solid (750 mg, 80%). MS-ESI: [M+H]+ 269.3 Example 205c tert-Butyl 2-Amino-6,7-dihydropyrazolo[1,5-a]pyrazine-5(4H)- carboxylate 205c 5 A 100-mL -neck round-bottomed flask was purged with nitrogen and d with 205b (0.75 g, 2.80 mmol), 10% palladium on carbon (50% wet, 280 mg), and methanol (30 mL). The mixture was evacuated, charged with hydrogen gas, and stirred at room temperature for 2 h. The hydrogen was then evacuated and nitrogen was charged into the flask. The catalyst was removed by filtration through a pad of ® and the filtrate was 10 concentrated under reduced pressure to afford 205c (524 mg, 79%). MS-ESI: [M+H]+ 239.1 Example 205d tert-Butyl 2-(5-Bromomethyloxo-1,2-dihydropyridin ylamino)-6,7-dihydropyrazolo[1,5-a]pyrazine-5(4H)-carboxylate 205d A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 205c (524 mg, 2.2 mmol), Pd2(dba)3 (201 mg, 0.22 mmol), 15 XantPhos (254 mg, 0.44 mmol), cesium carbonate (1434 mg, 4.4 mmol), and oxane (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 4 h.
After this time the reaction was cooled to room temperature. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified by silica-gel column chromatography eluting with 50:1 dichloromethane/methanol to afford 205d (600 mg, 70%) 20 as a yellow solid. MS-ESI: [M+H]+ 424.2 Example 205e utyl 2-(5-(3-(Acetoxymethyl)(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyloxo-1,2-dihydropyridin ylamino)-6,7-dihydropyrazolo[1,5-a]pyrazine-5(4H)-carboxylate 205e A sealed tube equipped with a magnetic stirrer was d with 205d (213 mg, 0.50 25 mmol), 3-(acetoxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- yl)pyridinylboronic acid 113i (192 mg, 0.50 mmol), Pd(dppf)Cl2 (41 mg, 0.050 mmol), sodium acetate (82 mg, 1.0 mmol), K3PO4 (212 mg, 1.0 mmol), itrile (10 mL), and water (0.5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 2 h. It was then filtered and the te was evaporated in vacuo. The residue was ed 30 by silica-gel column chromatography eluting with 25:1 dichloromethane/methanol to afford 205e (280 mg, 82%) as a yellow solid. MS-ESI: [M+H]+ 683.3 Example 205f (4-(1-Methyloxo(4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinylamino)-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)pyridinyl)methyl Acetate 205f 287 A mixture of 205e (280 mg, 0.41 mmol), 4.0 M HCl/dioxane (4 mL), and dichloromethane (4 mL) was stirred at room temperature for 2 h. It was then concentrated under reduced pressure to afford 205f as a yellow solid (165 mg, 66%), which was used for the next step without further purification. MS-ESI: [M+H]+ 583.3. 5 Example 205 2-[3-(hydroxymethyl)[1-methyloxo(4,5,6,7- ydropyrazolo[1,5-a]pyrazinylamino)pyridyl]pyridyl]-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indolone 205 A mixture of 205f (165 mg, 0.28 mmol) and lithium hydroxide (67 mg, 2.80 mmol) in i-propanol/THF (1:1, 4 mL) and water (1 mL) was stirred at room temperature for 1 h. The 10 mixture was evaporated in vacuo and diluted with water (4 mL). It was then extracted with ethyl acetate (10 mL X 2). The ed ethyl acetate extract was concentrated under d pressure and the residue was purified by reverse-phase prep-HPLC to afford 205 (70 mg, 46%) as a white solid. MS-ESI: [M+H]+ 541.2. 1H NMR (500 MHz, CDCl3) δ 8.48 (d, J = 5.0 Hz, 1H), 7.96 (d, J = 2.0 Hz, 1H), 7.72 (d, J = 2.0 Hz, 1H), 7.43 (s, 1H), 7.35 (d, J = 15 5.5 Hz, 1H), 6.90 (s, 1H), 5.70 (s, 1H), 5.01 (s, 1H), 4.64-4.61 (m, 1H), 4.50 (s, 1H), 4.34 (s, 1H), 4.16-3.99 (m, 6H), .87 (m, 1H), 3.71 (s, 3H), 3.30 (t, J = 5.5 Hz, 2H), 2.63-2.57 (m, 4H), 1.92-1.89 (m, 2H), 1.80-1.78 (m, 3H).
Example 206a tert-Butyl 3-Iodoazetidinecarboxylate 206a 288 HO O NBoc PPh3/I2/tol. NBoc O N NO2 Pd/C BocN BocN HO 100 °C I N NO2 MeOH, H2 N NH2 206a 206b 206c Br O O O ClHHN BocN N NH N N NH Br O HCHO, HOAc O HCl / 1,4-dioxane N NaBH(OAc)3 Xantphos, Pd2dba3 N 80 °C Br Cs2CO3, 1,4-dioxane Br 110 °C, 2 h 206d 206e AcO N OH O O N B OH N N O N N NH N NH OAc O O 113i N N N N Br Pd(dppf)Cl2 (0.05 eq), K3PO4 (2 eq), H2O O N NaOAC 3H2O(2 3CN, 100 oC, 2h 206g 206f A solution of tert-butyl oxyazetidinecarboxylate (3.5 g, 0.020 mol) in toluene (200 mL) was treated with imidazole (4.08 g, 0.060 mol), triphenylphosphine (0.60 g, 0.040 mol), and iodine (7.62 g, 0.030 mol). The mixture was heated at 100oC for 1 h. It was 5 then cooled to room temperature and poured into saturated NaHCO3 solution (30 mL). Excess triphenylphosphine was destroyed by addition of iodine until iodine tion persisted in organic layer. The mixture was washed with 5% Na2SO3 solution, dried over Na2SO4, and evaporated in vacuo. The residue was purified by silica-gel column chromatography to afford 206a (5.31 g, 93%). : [M+H]+ 284. 10 Example 206b tert-Butyl 3-(6-Nitropyridinyloxy)azetidinecarboxylate 206b A mixture of 206a (2.24 g, 7.9 mmol), 6-nitropyridinol (1.0 g, 7.2 mmol), and Cs2CO3 (2.6 g, 7.9 mmol) in DMF (8 mL) was heated at 125 oC in a sealed tube overnight.
The solid was filtered and washed with ethyl acetate (2 X 20 mL). The combined filtrate was 15 evaporated in vacuo and the residue was purified on reverse-phase Combiflash to afford 206b (1.25 g, 59%). MS-ESI: [M+H]+ 296. e 206c tert-Butyl 3-(6-Aminopyridinyloxy)azetidinecarboxylate 206c 289 A 100-mL Parr hydrogenation bottle was purged with nitrogen and charged with 206b (1.07 g, 3.6 mmol), 10% palladium on carbon (50% wet, 0.30 g), and methanol (60 mL). The bottle was evacuated, charged with hydrogen gas to a pressure of 25 psi, and shaken for 2 h on a Parr enation apparatus. The hydrogen was then evacuated and nitrogen charged to 5 the bottle. The catalyst was d by filtration through a pad of CELITE® and the te was concentrated under reduced pressure to afford 206c (0.95 g, 99%). MS-ESI: [M+H]+ 266.
Example 206d tert-Butyl 3-(6-(5-Bromomethyloxo-1,2-dihydropyridin- 3-ylamino)pyridinyloxy)azetidinecarboxylate 206d A 100-mL bottomed flask equipped with a reflux condenser was charged with 10 206c (950 mg, 3.6 mmol), XantPhos (125 mg, 0.29 mmol), Pd2dba3 (260 mg, 0.29 mmol), 3,5-dibromomethylpyridin-2(1H)-one (1.03 g, 3.9 mmol), Cs2CO3 (1.8 g, 7.2 mmol), and 1,4-dioxane (20 mL). The system was ted and refilled with N2. It was then heated at reflux for 2 h. After the completion of the reaction, the mixture was filtered off and washed with ol (100 mL). The combined filtrate was evaporated in vacuo and the residue was 15 purified on reverse-phase Combiflash to afford 206d (1.46 g, 90%). MS-ESI: [M+H]+ 451.
Example 206e Azetidinyloxy)pyridinylamino)bromo methylpyridin-2(1H)-one Hydrochloride 206e A mixture of 206d (1.46 g, 3.2 mmol) and HCl/1,4-dioxane (3.2 mL, 4M, 12.8 mmol) in methanol (20 mL) was heated at 80ºC for 1 h. The mixture was then concentrated under 20 reduced pressure to afford 206e (1.24 g, 99%). MS-ESI: [M+H]+ 351.
Example 206f 5-Bromomethyl(5-(1-methylazetidinyloxy)pyridin ylamino)pyridine-2(1H)-one 206f A mixture of 206e (1.24 g, 3.2 mmol), 37% aqueous formaldehyde on (15 mL,), acetic acid (1 mL), and NaBH(OAc)3 (1.36 g, 6.4 mmol) in methanol (10 mL) was stirred at 25 room temperature for 4 h. The solvent was evaporated in vacuo and the residue was extracted with ethyl acetate (3 X 20 mL). The combined extract was dried over sodium sulfate and trated under reduced re. The residue was purified on reverse-phase Combiflash to afford 206f (940 mg, 80%). MS-ESI: [M+H]+ 365.
Example 206g (4-(1-Methyl(5-(1-methylazetidinyloxy)pyridin 30 ylamino)oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)pyridinyl)methyl Acetate 206g A 100-mL round-bottomed flask equipped with a reflux condenser was charged with 206f (108 mg, 0.30 mmol), 3-(acetoxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)pyridinylboronic acid 113i (115 mg, 0.30 mmol), Pd(dppf)Cl2 (15 mg, 290 0.015 mmol), K3PO4 (135 mg, 0.60 mmol), sodium acetate rate (90 mg, 0.60 mmol) in itrile (10 mL) and water (0.5 mL). The system was evacuated and refilled with N2. The reaction mixture was heated at 100ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced re and the resulting residue was 5 purified by silica-gel column chromatography eluting with 25:1 dichloromethane/methanol to afford 206g (90 mg, 52%) as a yellow brown solid. MS-ESI: [M+H]+ 624.2.
Example 206 2-[3-(hydroxymethyl)[1-methyl[[5-(1-methylazetidinyl)oxy pyridyl]amino]oxopyridyl]pyridyl]-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indolone 206 10 A mixture of 206g ( 93.6 mg, 0.15 mmol) and lithium hydroxide (65 mg, 1.5 mmol) in THF/i-propanol (5:3, 8 mL) and water (2 mL) was stirred at 30oC for 1 h. The mixture was evaporated in vacuo and diluted with water (3 mL). It was then extracted with ethyl acetate (2 X 10 mL). The combined ethyl acetate extract was concentrated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 206 (35 mg, 42 %) as white 15 solid. MS-ESI: [M+H]+ 582.3. 1H NMR (500 MHz, CHCl3) δ 8.62 (d, J = 1.5 Hz, 1H), 8.51 (d, J = 5.0 Hz, 1H), 7.86 (d, J = 2.0 Hz, 1H), 7.84 (s, 1H), 7.76 (d, J = 3.0 Hz, 1H), 7.37 (d, J = 5.0 Hz, 1H), 7.12-7.10 (m, 1H), 6.90 (s, 1H), 6.81-6.80 (m, 1H), 5.07-5.04 (m, 1H), 4.77 (t, J = 5.5 Hz, 1H), 4.64-4.62 (m, 1H), 4.52-4.50 (m, 1H), 4.33-4.30 (m, 1H), .10 (m, 2H), 3.97-3.88 (m, 3H), 3.72 (s, 3H), 3.25-3.24 (m, 2H), 2.63-2.57 (m, 4H), 2.51 (s, 3H), 1.93- 20 1.91 (m, 2H), 1.80-1.79 (m, 2H).
Example 207a (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}[1-methyl({5-[(1-methylazetidinyl)oxy]pyridinyl}amino) oxopyridinyl]pyridinyl)methyl Acetate 207a 25 A 50-mL single-neck round-bottomed flask equipped with a reflux condenser was charged with 5-bromomethyl(5-(1-methylazetidinyloxy)pyridinylamino)-pyridin- 2(1H)-one 206f (108 mg, 0.40 mmol), cetyloxy)methyl]{4,4-dimethyloxo-1,10- diaza-tricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (240 mg, 0.60 mmol), f)Cl2 (20 mg, 0.02 mmol), K3PO4 (180 mg, 0.80 mmol), sodium acetate 291 trihydrate (120 mg, 0.80 mmol), water (0.5 mL), and acetonitrile (10 mL). The system was evacuated and refilled with N2. The reaction mixture was heated at 100ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 10:1 5 dichloromethane/methanol to afford 207a (100 mg, 45%) as a yellow brown solid. I : [M+H]+ 638.4.
Example 207 3-[3-(hydroxymethyl)[1-methyl[[5-(1-methylazetidin yl)oxypyridyl]amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 207 10 A mixture of 207a ( 90 mg, 0.15 mmol) and lithium hydroxide (65 mg, 1.5 mmol) in THF/ i-propanol(5:3, 8 mL) and water (2 mL) was stirred at 30oC for 1 h. The e was evaporated under d pressure and diluted with water (4 mL). It was then extracted with ethyl acetate (2 X 20 mL). The combined ethyl acetate extract was concentrated under reduced pressure and the residue was ed by reverse-phase prep-HPLC to afford 207 (30 15 mg, 38%) as white solid. LCMS: [M+H]+ 596.3. 1H NMR (500 MHz, CDCl3) δ 8.61 (d, J=2.0 Hz, 1H), 8.51 (d, J=5.0 Hz, 1H), 7.86-7.83 (m, 2H), 7.77 (d, J=3.0 Hz, 1H), 7.37 (d, J=5.0 Hz, 1H), 7.12-7.10 (m, 1H), 6.85 (s, 1H), 6.81 (d, J=3.5 Hz, 1H), 5.07-5.04 (m, 1H), 4.74-4.64 (m, 2H), 4.52-4.51 (m, 1H), 4.34-4.32 (m, 1H), 4.17-4.16 (m, 2H), 3.88-3.87 (m, 3H), 3.72 (s, 3H), .16 (m, 2H), 2.58 (d, J=5.5 Hz, 2H), 2.52 (s, 2H), 2.45 (s, 3H), 1.28 20 (s, 6H).
Example 208a 5-Ethylnitro-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine 208a 292 A 150-mL single-neck round-bottomed flask equipped with a ic r was charged with methanol (60 mL), 2-nitro-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine 205a (1.5 g, 8.9 mmol), ZnCl2 (2.43 g, 17.8 mmol), dehyde (784 mg, 17.8 mmol), and NaBH3CN 5 (1.12 g, 17.8 mmol). The mixture was stirred at room temperature for 2 h and concentrated under reduced pressure. The e was purified with silica-gel column chromatography eluting with 40:1 petroleum ether/ethyl acetate to afford 208a (1.4 g, 81%) as a yellow oil.
MS-ESI: [M+H]+ 197 Example 208b 5-Ethyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinamine 10 208b A 50-mL single-neck round-bottomed flask was purged with nitrogen and charged with 208a (1.4 g, 7.1 mmol), 10% palladium on carbon (50% wet, 208 mg), methanol (30 mL), and hydrogen gas. The mixture was stirred at room ature under en atmosphere for 2 h. The catalyst was removed by filtration through a pad of CELITE® and 15 the filtrate was concentrated under reduced pressure to afford 208b (1.0 g, 84%) as yellow oil.
MS-ESI: [M+H]+ 167 Example 208c 5-Bromo(5-ethyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin- 2-ylamino)methylpyridin-2(1H)-one 208c A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 20 reflux condenser was charged with 208b (1.0 g, 6.0 mmol), 3,5-dibromomethylpyridin- 2(1H)-one (1.6 g, 6.0 mmol), Pd2(dba)3 (274 mg, 0.30 mmol), XantPhos (347 mg, 0.60 293 mmol), cesium carbonate (3.9 g, 12.0 mmol), and 1,4-dioxane (50 mL). After three cycles of vacuum/argon flush, the mixture was stirred at 100 oC for 3 h. It was then filtered and the filtrate was evaporated in vacuo. The residue was ed by silica-gel column chromatography eluting with 40:1 dichloromethane/methanol to afford 208c (630 mg, 29%) 5 as a yellow solid. MS-ESI: [M+H]+ 352 Example 208d 3-(5-Ethyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin ylamino)methyl(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 208d A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 1,4-dioxane (20 mL), 208c (350 mg. 0.99 mmol), bis 10 (pinacolato) diboron (1.31 g, 4.99 mmol), Pd2(dba)3 (45 mg, 0.050 mmol), X-phos (58 mg, 0.10 mmol), and ium acetate (291 mg, 2.97 mmol). After ng nitrogen through the mixture for 30 minutes, it was heated at 90 oC for 3 h. Then it was filtered and the filtrate was ated in vacuo. The residue was washed with petroleum ether to afford 208d (120 mg, 30%) as a brown solid. MS-ESI: [M+H]+ 400.2 15 Example 208e 4-(5-(5-Ethyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin ylamino)methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrido [3,4-b]indolizin-2(1H)-yl)nicotinaldehyde 208e A 50-mL single-neck bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 208d (120 mg, 0.30 mmol), 4-chloro(1-oxo- 20 3,4,6,7,8,9-hexahydropyrido-[3,4-b]indolizin-2(1H)-yl)nicotinaldehyde 139a (99 mg, 0.30 mmol), PdCl2(dppf) (13 mg, 0.015 mmol), K3PO4 (127 mg, 0.60 mmol), sodium acetate (49 mg, 0.60 mmol), acetonitrile (10 mL), and water (0.5 mL). The system was evacuated and ed with N2. The reaction mixture was heated at 100ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the 25 resulting residue was purified by silica-gel column tography eluting with 30:1 dichloromethane/methanol to afford the 208e (95 mg, 56%) as a yellow solid. MS-ESI: [M+H]+ 567.2.
Example 208 5-[(5-ethyl-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazinyl)amino]- 1-methyloxopyridyl](hydroxymethyl)pyridyl]-3,4,6,7,8,9-hexahydropyrido[3,4- 30 b]indolizinone 208 To a mixture of 208e (95 mg, 0.16 mmol) at 0 oC in methanol (10 mL) was added sodium borohydride (19 mg, 0.50 mmol). The reaction mixture was stirred for 30 minutes and quenched with water (2.0 mL). It was then trated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 208 (8 mg, 9%) as white solid. 294 MS-ESI: [M+H]+ 569.3. 1H NMR (500 MHz, CDCl3) δ 8.46 (d, J = 5.0 Hz, 1H), 7.92 (d, J = 2.0 Hz, 1H, 7.70 (d, J = 2.0 Hz, 1H), 7.40 (s, 1H), 7.30 (d, J = 5.0 Hz, 1H), 6.30 (s, 1H), 5.71 (s, 1H), 4.93-4.96 (m, 1H), 4.63-4.61 (m, 1H), 4.42-4.26 (m, 2H), 4.09 (s, 2H), 3.94-3.81 (m, 3H), 3.69-3.68 (m, overlap, 5H), 3.06-2.90 (m, 4H), 2.81 (d, J = 3.0 Hz, 2H), 2.66 (d, J = 3.5 5 Hz, 2H), 2.04-2.00 (m, 2H), .85 (m, 2H), 1.20 (t, J = 7.5 Hz, 3H).
Example 209a 1-(2-Nitro-6,7-dihydropyrazolo[1,5-a]pyrazin-5(4H)- yl)ethanone 209a Br O O O O N K2CO3, N HN Pd/C, MeOH N N Br CH3COCl rt, 2 h N N N N N NH CH2Cl2, N Pd2(dba)3 N NO2 NO2 N NH2 O rt, overnight XantPhos 205a 209a 209b Cs2CO3 N dioxane Br 100 oC 209c O OAc O N N N N Cl N Pd2(dba)3 N O N N NH X-Phos N NH 113h OAc O O N AcOK N N dioxane O N Pd(dppf)Cl2 B 100 oC K3PO4, NaOAc, O N O CH3CN/H2O 100 oC 209e 209d To a solution of 2-nitro-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine 205a (672 mg, 4.0 10 mmol) in dichloromethane (20 mL) was added acetyl chloride (936 mg, 12.0 mmol) and K2CO3 (1104 mg, 8.0 mmol). The mixture was stirred overnight. It was then filtered and the filtrate was concentrated under reduced re. The resulting residue was purified by silicagel column chromatography eluting with 100:1 romethane/methanol to afford 209a as white solid (500 mg, 60%). MS: [M+H]+ 211.2 15 Example 209b 1-(2-Amino-6,7-dihydropyrazolo[1,5-a]pyrazin-5(4H)- yl)ethanone 209b A 50-mL single-neck round-bottomed flask was purged with nitrogen and charged with 209a (492 mg, 2.34 mmol), 10% palladium on carbon (50% wet, 234 mg), and methanol (20 mL). The mixture was evacuated, d with hydrogen gas, and stirred at room 20 temperature for 2 h. The hydrogen was then evacuated and nitrogen was charged into the flask. The st was removed by filtration through a pad of ® and the filtrate was concentrated under reduced pressure to afford 209b (380 mg, 80%). MS: [M+H]+ 181.1 295 Example 209c cetyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin ylamino)bromomethylpyridin-2(1H)-one 209c A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 3,5-dibromomethylpyridin-2(1H)-one (481 mg, 1.8 5 mmol), 209b (270 mg, 1.5 mmol), oxane (20 mL), Pd2(dba)3 (137 mg, 0.15 mmol), XantPhos (173 mg, 0.30 mmol), and cesium carbonate (978 mg, 3.0 mmol). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 6 h. After this time the reaction was cooled to room temperature. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified by silica-gel column chromatography eluting with 50:1 10 dichloromethane/methanol to afford 209c (540 mg, 89%) as a yellow solid. MS: [M+H]+ 368.0 Example 209d 3-(5-Acetyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin ylamino)methyl(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 209d A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 15 reflux condenser was charged with 209c (365 mg, 1.0 mmol), Pin2B2 (1.26 g, 5.0 mmol), Pd2(dba)3 (91 mg, 0.10 mmol), X-phos (92 mg, 0.20 mmol), AcOK (294 mg, 3.0 mmol), and dioxane (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at 60ºC for 16 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica-gel column chromatography 20 eluting with 50:1 methylene de/methanol to afford 209d as a brown solid (330 mg, 80%). MS: [M+H]+ 414.2 Example 209e (5-Acetyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin ylamino)methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl Acetate 209e 25 A sealed tube equipped with a magnetic stirrer was charged with 209d (185 mg, 0.50 mmol), (4-chloro(1-oxo-3,4,6,7,8,9-hexahydropyrazino-[1,2-a]indol-2(1H)-yl)pyridine yl)methyl acetate 113h (192 mg, 0.50 mmol), Pd(dppf)Cl2 (41 mg, 0.050 mmol), sodium acetate (82 mg, 1.0 , K3PO4 (212 mg, 1.0 mmol), water (0.5 mL), and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 2 h. It 30 was then ed and the filtrate was evaporated in vacuo. The e was purified by silicagel column chromatography eluting with 25:1 dichloromethane/methanol to afford 209e (150 mg, 48%) as a yellow solid. MS-ESI: [M+H]+ 625.4 296 Example 209 2-[4-[5-[(5-acetyl-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazinyl)amino]- 1-methyloxopyridyl](hydroxymethyl)pyridyl]-3,4,6,7,8,9-hexahydropyrazino[1,2- lone 209 A mixture of 209e (150 mg, 0.24 mmol) and m hydroxide (58 mg, 2.4 mmol) in 5 i-propanol/THF (1:1, 4 mL) and water (1 mL) was stirred at room temperature for 1 h. The mixture was evaporated in vacuo and diluted with water (4 mL). It was then extracted with ethyl acetate (2 X 10 mL). The ed ethyl acetate t was concentrated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 209 (75 mg, 53%) as a white solid. : [M+H]+ 583.3. 1H NMR (500 MHz, T=80oC, DMSO-d6) 10 δ 8.44 (d, J = 8.5 Hz, 1H), 7.93-7.90 (m, 2H), 7.34 (d, J = 4.5 Hz, 1H), 7.29 (d, J = 8.5 Hz, 1H), 6.56 (s, 1H), 5.98 (s, 1H), 4.72-4.63 (m, 3H), 4.45-4.43 (m, 2H), 4.16-4.10 (m, 3H), 3.99-3.86 (m, overlap, 5H), 3.58 (s, 3H), 2.62-2.57 (m, 2H), 2.49-2.47 (m, 2H), 2.08 (s, 3H), 1.83-1.77 (m, 2H), 1.72-1.68 (m, 2H).
Example 210a 4-Chloro{4,4-dimethyloxo-1,10-diazatricyclo- 15 [6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarboxylic Acid 210a O N HO O NaClO2, NaH2PO4 N N Cl N Cl tert-butyl alcohol, CH2Cl2 O N H2O, -10 °C, o/n O N 108a 210a O N N O N NH N O N O N B N NH O O OH O N 191j N N PdCl2(dppf), K3PO4,AcONa O N CH3CN/H2O, 100 °C, 3h 210b To a mixture of 4-chloro{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}pyridinecarbaldehyde 108a (500 mg, 1.46 mmol), tert-butyl alcohol (20 mL), and dichloromethane (5 mL) was added 2-methylbutene (3066 mg, 43.8 mmol). An 20 aqueous solution (8 mL) of NaClO2 (263 mg, 2.92 mmol) and NaH2PO4·2water (683 mg, 4.38 mmol) was added dropwise at -10oC and the reaction e was stirred at -10 oC for 297 overnight. It was concentrated under reduced pressure and the residue was extracted with ethyl acetate (4 × 20 mL). The combined organic extract was dried over MMggSSOO44 and concentrated. The residue was purified with reverse-phase prep-HPLC to afford 210a (315 mg, 60%) as a pale yellow solid. MS-ESI: [M+H]+ 360.1 5 Example 210b 2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl} [1-methyl({5-[(2S)methyl(oxetanyl)piperazinyl]pyridin- mino)oxo-1,6-dihydropyridinyl]pyridinecarboxylic Acid 210b A 25-mL round-bottomed flask equipped with a reflux condenser was charged with 210a (400 mg, 1.1 mmol), (S)methyl(5-(2-methyl(oxetanyl)piperazin 10 idinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 191j (536 mg, 1.1 mmol), PdCl2(dppf) (81 mg, 0.11 mmol), K3PO4 (466 mg, 2.2 mmol), sodium acetate (216 mg, 2.2 mmol), acetonitrile (10 mL), and water (0.2 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 3 h. It was then ed and the filtrate was evaporated in vacuo. The residue was purified by silica-gel column 15 chromatography eluting with 1:3 petroleum/ethyl acetate to afford 210b as a yellow solid (306 mg, 41%). MS-ESI: [M+H]+ 679.3 Example 210 2-(7,7-dimethyloxo-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- b]pyrazinyl)[1-methyl[[5-[(2S)methyl(oxetanyl)piperazinyl] l]amino]oxopyridyl]pyridinecarboxamide 210 20 A 25-mL round-bottomed flask was charged with 210b (300 mg, 0.44 mmol), triethylamine (1 mL), DMAP (5 mg, 0.040 mmol), HATU (250 mg, 0.66 mmol), and DMF (10 mL). The mixture was stirred at room temperature for 0.5 h. Then 37% aqueous ammonia (15 mL) was added slowly and the reaction was d at room temperature for another 2.5 h.
The mixture was treated with 20 mL water and extracted with dichloromethane (3 X 20 mL). 25 The combined organic extract was concentrated under reduced pressure and residue was purified with e-phase prep-HPLC to afford 210 (98 mg, 33%) as yellow solid. MS-ESI: [M+H]+ 678.3. 1H NMR (500 MHz, DMSO) δ 8.71 (d, J = 2.0 Hz, 1H), 8.55 (d, J = 2.5 Hz, 1H), 8.41 (s, 1H), 7.84 (d, J = 3.0 Hz, 1H), 7.61 (s, 1H), 7.49 (s, 1H), 7.45-7.42 (m, 2H), 7.38-7.36 (m, 1H), 7.24-7.22 (m, 1H), 6.49 (s, 1H), .54 (m, 2H), .47 (m, 1H), 30 4.43-4.40 (m, 1H), 4.12-4.11 (m, 2H), 4.04-4.00 (m, 2H), 3.67-3.66 (m, 1H), 3.57 (s, 3H), .37 (m, 1H), 3.10-3.08 (m, 1H), 2.97-2.92 (m, 1H), 2.55-2.53 (m, 3H), 2.41 (s, 2H), 2.36-2.29 (m, 2H), 2.21-2.18 (m, 1H), 1.21 (s, 6H), 0.93 (d, J = 6.0 Hz, 3H) 298 Example 211 2-(7,7-dimethyloxo-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- b]pyrazinyl)-N-methyl[1-methyl[[5-[(2S)methyl(oxetanyl)piperazinyl]- 2-pyridyl]amino]oxopyridyl]pyridinecarboxamide 211 A round-bottomed flask was charged with 2-{4,4-dimethyloxo-1,10-diazatricyclo 5 [6.4.0.02,6]dodeca-2(6),7-dienyl}[1-methyl({5-[(2S)methyl(oxetan yl)piperazinyl]pyridinyl}amino)oxo-1,6-dihydropyridinyl]pyridinecarboxylic acid 210b (300 mg, 0.44 mmol), ylamine (1 mL), DMAP (5 mg, 0.040 mmol), HATU (250 mg, 0.66 mmol), and DMF (10 mL). The mixture was stirred at room temperature for 0.5 h. Then CH3NH2 (27 mg, 0.88 mmol) was added slowly and the reaction was d at 10 room temperature for another 2.5 h. The e was treated with water (20 mL) and ted with dichloromethane (20 mL X 3). The combined organic extract was concentrated under reduced pressure and the residue was purified with e-phase prep-HPLC to afford 211 (106 mg, 35%) as yellow solid. MS-ESI: [M+H]+ 692.5. 1H NMR (500 MHz, DMSO-d6) δ 8.71 (d, J = 2.5 Hz, 1H), 8.55 (d, J = 5.0 Hz, 1H), 8.42 (s, 1H), 8.11-8.08 (m, 1H), 7.86 (d, 15 J = 2.5 Hz, 1H), 7.44-7.41 (m, 2H), 7.38-7.35 (m, 1H), 7.24-7.22 (m, 1H), 6.48 (s, 1H), 4.58- 4.56 (m, 2H), .46 (m, 1H), 4.43-4.41 (m, 1H), 4.08-4.07 (m, 2H), 3.97-3.94 (m, 2H), 3.66-3.65 (m, 1H), 3.58 (s, 3H), 3.41-3.39 (m, 1H), 3.10-3.08 (m, 1H), 2.97-2.93 (m, 1H), 2.56 (s, 2H), 2.53-2.48 (m, overlap, 4H), 2.37-2.36 (m, 2H), 2.35-2.31 (m, 2H), 2.29-2.19 (m, 1H), 1.21 (s, 6H), 0.93 (d, J = 6.0 Hz, 3H). 20 Example 212a {4-[5-({5-Acetyl-4H,6H,7H-pyrazolo[1,5-a]pyrazin yl}amino)methyloxopyridinyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]-dodeca-2(6),7-dienyl}pyridinyl}methyl Acetate 212a A 50-mL round-bottomed flask equipped with a reflux condenser was charged with 3- 25 (5-acetyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinylamino)bromomethylpyridin- 2(1H)-one 209c(185 mg, 0.50 mmol), {3-[(acetyloxy)methyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (200 mg, 0.50 mmol), Pd(dppf)Cl2 (41 mg, 0.050 mmol), sodium acetate (82 mg, 1.0 mmol), K3PO4 299 (212 mg, 1.0 mmol), water (0.5 mL), and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 2 h. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified by silica-gel column chromatography eluting with 25:1 dichloromethane/methanol to afford 212a (180 mg, 56%) 5 as a yellow solid. MS-ESI: [M+H]+ 639.3 e 212 3-[4-[5-[(5-acetyl-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazinyl)amino]- 1-methyloxopyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 212 A mixture of 212a (180 mg, 0.28 mmol) and lithium hydroxide (67 mg, 2.8 mmol) in 10 i-propanol/THF (1:1, 4 mL) and water (1 mL) was stirred at 30oC for 1 h. The mixture was evaporated in vacuo and diluted with water (4 mL). It was then extracted with ethyl acetate (2 X 10 mL). The combined ethyl acetate extract was concentrated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 212 (70 mg, 42%) as a white solid. MS-ESI: [M+H]+ 597.3. 1H NMR (500 MHz, T=80oC, DMSO-d6) δ 8.47 (d, J = 8.0 15 Hz, 1H), 7.95 (d, J = 4.0 Hz, 1H), 7.92 (s, 1H), 7.37 (d, J = 3.5 Hz, 1H), 7.31 (d, J = 8.5 Hz, 1H), 6.57 (s, 1H), 6.00 (s, 1H), 4.66 (bs, 2H), 4.47 (s, 2H), .18 (m, 3H), 4.00-3.99 (m, 3H), 3.92-3.88 (m, 3H), 3.61 (s, 3H), 2.59 (s, 2H), 2.46 (s, 2H), 2.11 (s, 3H), 1.25 (s, 6H).
Example 213a omethyl(2-methylpyrimidinylamino)pyridin-2(1H)- one 213a O N N N N N Cl N NH N NH N NH F O O N O O O N N 134a O N N N B Br F O O N 213a 213b pd(dppf)Cl2, K3PO4, 213c NaOAc, 2O, 20 reflux, 2.5 h Following the procedures described in Example 196, reaction of 2-methylpyrimidin- 4-amine (2.0 g, 18.3 mmol) and 3,5-dibromomethylpyridin-2(1H)-one (9.6 g, 36 mmol) afforded 213a as a yellow solid (2.3 g, . MS: [M+H]+ 295. 1H NMR (500 MHz, DMSO-d6) δ 9.20 (s, 1H), 8.78 (s, 1H), 8.26 (d, J = 4.5 Hz, 1H), 7.68 (s, 1H), 7.18 (d, J = 25 4.5 Hz, 1H), 3.59 (s, 3H), 2.52 (s, 3H). e 213b 1-Methyl(2-methylpyrimidinylamino)(4,4,5,5- tetramethyl-1,3,2- dioxaborolanyl)pyridin-2(1H)-one 213b A 100-mL single-neck round-bottomed flask equipped with a ic stirrer and a reflux condenser was charged with bis (pinacolato) diboron (689 mg, 2.61 mmol), 1,4- 300 e (30 mL), 213a (307 mg, 1.04 mmol), Pd2(dba)3 (47 mg, 0.050 mmol), X-phos (48 mg, 0.10 mmol), and ium acetate (305 mg, 3.12 mmol). The mixture was heated at 65oC for 6 h. It was then ed and the filtrate was evaporated in vacuo to afford 213b (300 mg, 84%) as a brown solid. MS: [M+H]+ 342.2 5 Example 213c 2-(10-Fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl)(1-methyl(2-methylpyrimidinylamino)oxo-1,6-dihydropyridin yl)nicotinaldehyde 213c A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 4-chloro(10-fluorooxo-3,4,6,7,8,9- 10 hexahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 134c (150 mg, 0.43 mmol), 213b (147 mg 0.43 mmol), Pd(dppf)Cl2 (35 mg, 0.043 mmol), sodium acetate (71 mg, 0.86 mmol), K3PO4 (182 mg, 0.86 mmol), water (0.5 mL), and acetonitrile (15 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 2.5 h. After cooling to room temperature the reaction was filtered. The filtrate was concentrated under reduced pressure 15 and the resulting residue was purified by silica-gel column chromatography eluting with 40:1 dichloromethane/methanol to afford 213c as a yellow solid (130 mg, 57%). MS-ESI: [M+H]+ 528.2.
Example 213 oro[3-(hydroxymethyl)[1-methyl[(2-methylpyrimidin no]oxopyridyl]pyridyl]-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indolone 213 20 To a solution of 213c (120 mg, 0.23 mmol) at 0 oC in methanol (10 mL) was added sodium borohydride (26 mg, 0.69 mmol). The reaction mixture was stirred for 20 minutes and quenched with water (10 mL). It was then extracted with dichloromethane (3 X 20 mL) and the combined organic layer was concentrated under reduced pressure. The residue was ed by reverse-phase prep-HPLC to afford 213 (62 mg, 44 %) as a white solid. MS-ESI: 25 [M+H]+ 530.3. 1H NMR (500 MHz, DMSO-d6) δ 9.11 (s, 1H), 8.93 (d, J = 2.5 Hz, 1H), 8.50 (d, J = 5.0 Hz, 1H), 8.21 (d, J = 6.0 Hz, 1H), 7.69 (d, J = 2.5 Hz, 1H), 7.37 (d, J = 5.0 Hz, 1H), 7.13 (d, J = 5.5 Hz, 1H), 4.96 (t, J = 5.5 Hz, 1H), 4.57-4.45 (m, 2H), 4.23-4.18 (m, 2H), 4.08-4.05 (m, 1H), 3.90-3.87 (m, 1H), 3.62 (s, 3H), .56 (m, 2H), 2.45 (s, 3H), 2.43- 2.42 (m, 2H), 1.78-1.76 (m, 2H), 1.72-1.66 (m, 2H) . 30 Example 214a 3-Bromoiodopyridinol 214a 301 AcO N N B(OH)2 I Br O N I Br 199e N O N OH Pd(dppf)Cl2, K3PO4, NaOAc, CH3CN, H2O 30 oC, 3 h 214a 214b O O S S Br N NH AcO O N NH2 OAc O N N N N N N Pd2(dba)3, xantphos, Cs2CO3 Dioxane, O N O N 100 °C, 10 h 214c 214d A 100-mL single-neck round-bottomed flask equipped with a magnetic r was charged with acetonitrile (50 mL), trifluoroacetic acid (10 mL), 3-bromopyridinol (4.0 g, 11.56 mmol) and N-iodosuccinimide (5.2 g, 11.56 mmol). The mixture was stirred at room 5 temperature for 15 h. The mixture was diluted with water (100 mL) and resulting white solid was collected by filtration to afford 214a (6.6 g, 96%) as a white solid. MS-ESI: [M+H]+ 300 Example 214b 3-Bromoiodomethylpyridin-2(1H)-one 214b A 100-mL -neck round-bottomed flask equipped with a magnetic stirrer was charged with DMF (50 mL), 214a (6.0 g, 20.0 mmol), iodomethane (4.26 g, 30.0 mmol), and 10 K2CO3 (5.52 g, 40.0 mmol). The mixture was stirred at room temperature for 2 h and diluted with water (200 mL). The resulting white solid was collected by tion to afford 214b (5.97 g, 95%) as a white solid. : [M+H]+ 314 Example 214c [4-(5-Bromomethyloxo-1,6-dihydropyridinyl){4,4- dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl]methyl 15 Acetate 214c A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with 214b (1.57 g, 5.0 mmol), {3-[(acetyloxy)methyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]-dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (1.98 g, 5.0 mmol), PdCl2(dppf) (205 mg, 0.25 mmol), K3PO4 (2.12 g, 10.0 mmol), Sodium acetate (820 20 mg, 10.0 mmol), acetonitrile (45 mL), and water (1 mL). The system was evacuated and ed with N2. The reaction mixture was stirred at 30oC for 3 h. It was then filtered and the te was concentrated under reduced pressure. The resulting residue was purified by gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 214c (580 mg, 22%) as a white solid. MS-ESI: [M+H]+ 539.2. 1H NMR (500 MHz, CDCl3) δ 8.49 (d, J 302 = 5.0 Hz, 1H), 7.84 (d, J = 2.5 Hz, 1H), 7.45 (d, J = 2.0 Hz, 1H), 7.09 (d, J = 5.0 Hz, 1H), 6.79 (s, 1H), 5.15 (s, 2H), 4.55-4.51 (m, 1H), 4.27-4.25 (m, 1H), .13 (m, 1H), 4.06- 4.04 (m, 1H), 3.68 (s, 3H), .56 (m, 2H), 2.51 (s, 2H), 1.86 (s, 3H), 1.28 (s, 6H).
Example 214d 4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 5 2(6),7-dienyl}[1-methyloxo({4H,6H,7H-pyrano[4,3-d][1,3]thiazolyl}amino)- 1,6-dihydro-pyridinyl]pyridinyl)methyl Acetate 214d A 50-mL -neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 1,4-dioxane (10 mL), 214c(150 mg, 0.28 mmol), Pd2(dba)3 (27 mg, 0.030 mmol), XantPhos (35 mg, 0.060 mmol), and cesium carbonate (183 10 mg, 0.56 mmol). After three cycles of vacuum/argon flash, the mixture was heated at 100oC for 10 h. After this time the reaction was cooled to room temperature. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified by silica-gel column chromatography eluting with 50:1 dichloromethane/methanol to afford 214d (89 mg, 52%) as a yellow solid. MS-ESI: [M+H]+ 615.2 15 Example 214 3-[4-[5-(6,7-dihydro-4H-pyrano[4,3-d]thiazolylamino)methyl oxopyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 214 A mixture of 214d (89 mg, 0.14 mmol), lithium hydroxide (35 mg, 1.45 mmol), and water/THF/i-propanol (3 mL /5 mL /5 mL) was stirred at 30oC for 2 h. The reaction mixture 20 was then concentrated under reduced pressure and the residue was extracted with dichloromethane (2 X 10 mL). The ed dichloromethane extract was concentrated under reduced pressure. The residue was purified with reverse-phase prep-HPLC to afford 214 (45 mg, 50%) as a yellow solid. MS-ESI: [M+H]+ 573.2. 1H NMR (500 MHz, DMSO-d6) δ 10.01 (s, 1H), 8.67 (d, J = 2.5 Hz, 1H), 8.50 (d, J = 5.0 Hz, 1H), 8.59 (d, J = 2.5 Hz, 1H), 25 7.34 (d, J = 5.0 Hz, 1H), 6.56 (s, 1H), .94 (m, 1H), 4.61 (s, 2H), 4.46-4.43 (m, 2H), 4.22-4.17 (m, 3H), 3.89-3.87 (m, 3H), 3.61 (s, 3H), 2.62-2.57 (m, 4H), 2.43 (s, 2H), 1.22 (s, 6H).
Example 215a 3-{4,4-Dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{1-methyl[(2-methylpyrimidinyl)amino]oxo-1,6- 30 dihydropyridinyl}pyridinecarbaldehyde 215a 303 N N NH O O N B N O N NH O N 213b O O N N Br N N O Pd(dppf)Cl2, K3PO4, N O NaOAc,CH3CN/H2O, N 100 °C, 4 h 107f 215a A sealed tube was charged with 3-bromo{4,4-dimethyloxo-1,10-diazatricyclo- [6.4.0.02,6]dodeca-2(6),7-dienyl}-pyridinecarbaldehyde 107f (210 mg, 0.54 mmol), 1- methyl(pyrimidinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin 5 one 213b (177 mg, 0.54 mmol), PdCl2(dppf) (42 mg, 0.050 mmol), K3PO4 (210 mg, 1.0 mmol), and sodium acetate (85 mg, 1.0 mmol), acetonitrile (8 mL), and water (0.5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 4 h. It was then filtered and the te was evaporated in vacuo. The residue was purified by silica-gel column chromatography eluting with 20:1 dichloromethane/methanol to afford 215a (150 mg, 10 53%). MS-ESI: [M+H]+ 524.2 .
Example 215 3-[4-(hydroxymethyl)[1-methyl[(2-methylpyrimidin yl)amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 215 A mixture of 215a (150 mg, 0.28 mmol) and NaBH4 (20 mg, 0.50 mmol) in methanol 15 (5 mL) was stirred at 25oC for 0.2 h. The mixture was ed by water (5 mL) and evaporated in vacuo. The residue was extracted with ethyl acetate (3 X 10 mL). The combined ethyl e t was concentrated under d pressure and the residue was purified with reverse-phase prep-HPLC to afford 215 (80 mg, 53%). MS-ESI: [M+H]+ 526.3. 1H NMR (500 MHz, DMSO-d δ 9.09 (s, 1H), 8.89 (s, 1H), 8.56 (s, 1H), 8.53 (s, 1H), 8.20 6) 20 (d, J = 4.0 Hz, 1H), 7.61(s, 1H), 7.13 (d, J = 6.0 Hz, 1H), 6.52 (s, 1H) , 5.19-5.18 (m, 1H), 4.47-4.46 (m, 2H), 4.23-4.20 (m, 3H), 3.95-3.93 (m, 1H), 3.62 (s, 3H), 2.57 (s, 2H), 2.42 (s, 3H), 2.41 (s, 2H), 1.21 (s, 6H).
Example 216a 1-Methyl(5-methyl-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin- 2-ylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 216a 304 5-Bromomethyl(5-methyl-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin ylamino)pyridin-2(1H)-one 203b (997 mg, 2.8 mmol) was dissolved in dioxane (50 mL), ed by addition of bis(pinacolato)diboron (3.0 g, 12.0 mmol), Pd2(dba)3 (128 mg, 5 ol), X-phos (134 mg, 0.28 mmol), and potassium acetate (823 mg, 8.4 mmol). After three cycles of vacuum/argon flush, the mixture was heated at 65oC for 2 h. The mixture was cooled to room temperature and filtered. The filtrate was trated under re and the residual was washed with petroleum ether (2 × 10 mL) to afford 216a as a yellow solid (968 mg, 86%), which was used in the next step without further purification. MS-ESI: [M+H]+ 10 403.2 Example 216b 4-[1-Methyl({5-methyl-4H,6H,7H-[1,3]thiazolo[5,4- c]pyridinyl}amino)oxopyridinyl]{6-oxothia-4,5- diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridinecarbaldehyde 216b A round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was 15 charged with ro{6-oxothia-4,5- diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trien- 5-yl}pyridinecarbaldehyde 124a (138 mg, 0.40 mmol), 216a (240 mg, 0.60 mmol), PdCl2(dppf) (20 mg, 0.020 mmol), K3PO4 (180 mg, 0.80 mmol), sodium acetate trihydrate (120 mg, 0.80 mmol), water (6 drops), and acetonitrile (15 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 2 h. Then, it was cooled to room 20 temperature and filtered. The filtrate was evaporated under reduced pressure and the residue was purified by silica-gel column chromatography eluting with 25:1 romethane/methanol to afford 216b as a yellow solid (100 mg, 45%). MS-ESI: [M+H]+ 586.2.
Example 216 3-[3-(hydroxymethyl)[1-methyl[(5-methyl-6,7-dihydro-4H- 25 thiazolo[5,4-c]pyridinyl)amino]oxopyridyl]pyridyl]-6,7,8,9- tetrahydrobenzothiopheno[2,3-d]pyridazinone 216 305 To a solution of 216b (100 mg, 0.15 mmol) in methanol (6 mL ) was added NaBH4 ( 18 mg, 0.45 mmol). The reaction mixture was stirred at 30oC for 1 h and quenched with brine (10 mL). It was then evaporated under reduced re. The residue was extracted with dichloromethane (2 X 20 mL) and the combined organic layer was concentrated under 5 reduced pressure. The resulting e was purified by e-phase prep-HPLC to afford 216 as a white solid (40 mg, 40%). MS-ESI: [M+H]+ 588.3. 1H NMR (500 MHz, CDCl3) δ 8.68 (d, J = 5.0 Hz, 1H), 8.43 (d, J = 2.0 Hz, 1H), 8.31 (s, 1H), 8.29 (s, 1H), 7.79 (d, J = 2.5 Hz, 1H), 7.55 (d, J = 5.0 Hz, 1H), 4.43-4.39 (m, 3H), 3.73 (s, 3H), 3.57-3.55 (m, 2H), 3.00- 2.98 (m, 2H), 2.88-2.86 (m, 2H), 2.82-2.80 (m, 4H), 2.50 (s, 3H), 2.03-1.95 (m, 4H). 10 Example 217a (4-(5-Bromomethyloxo-1,6-dihydropyridinyl)(1- oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl Acetate 217a Br O Br AcO N AcO N O N OH I N B 214b N N OH O N O N Pd(dppf)Cl2, K3PO4, H2O 113i NaOAc,CH3CN, rt, 5 h 217a H H N N N N N N NH2 NH AcO N O a)3, Xantphos, N N Cs2CO3, dioxane, 6 h 100oC O N 217b A sealed tube equipped with a magnetic stirrer was charged with 3-(acetoxymethyl)- 2-(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinylboronic acid 113i 15 (766 mg, 2.0 mmol), 3-bromoiodomethylpyridin-2(1H)-one 214b (626 mg, 2.0 mmol), Pd(dppf)Cl2 (164 mg, 0.20 mmol), sodium acetate (328 mg, 4.0 mmol), K3PO4 (848 mg, 4.0 mmol), acetonitrile (10 mL), and water (0.5 mL). After three cycles of vacuum/argon flush, the mixture was stirred at room temperature for 5 h. It was then filtered and the filtrate was evaporated in vacuo. The residue was ed by silica-gel column chromatography eluting 20 with 50:1 dichloromethane/methanol to afford 217a (700 mg, 67%) as a yellow solid. MSESI : [M+H]+ 525.2 306 Example 217b (4-(5-(1H-Imidazo[4,5-b]pyridinylamino)methyloxo- 1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- yl)pyridinyl)methyl Acetate 217b A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer and 5 reflux ser was charged with 217a (158 mg, 0.30 mmol), 1H-imidazo[4,5-b]pyridin amine (80 mg, 0.60 mmol), Pd2(dba)3 (27 mg, 0.030 mmol), XantPhos (35 mg, 0.061 mmol), cesium carbonate (200 mg, 0.60 mmol), and 1,4-dioxane (5 mL). After three cycles of vacuum/argon flush, the mixture was stirred at 100oC for 6 h. After this time the reaction was cooled to room temperature. It was then filtered and the filtrate was ated in vacuo. The 10 residue was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 217b (40 mg, 23%) as a yellow solid. MS-ESI: [M+H]+ 579.4 Example 217 2-[3-(hydroxymethyl)[5-(1H-imidazo[4,5-b]pyridinylamino) methyloxopyridyl]pyridyl]-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indolone 217 15 A mixture of 217b (40 mg, 0.070 mmol) and m ide (26 mg, 0.70 mmol) in anol/THF (1:1, 4 mL) and water (1 mL) was stirred at room temperature for 1 h. The mixture was evaporated in vacuo and the residue was diluted with water and ethyl e.
The water phase was separated and extracted with ethyl acetate (2 X 10 mL). The combined ethyl acetate extract was concentrated under reduced pressure and the residue was purified by 20 reverse-phase prep-HPLC to afford 217 (15 mg, 40%) as a white solid. MS-ESI: [M+H]+ 537.3. 1H NMR (500 MHz, DMSO-d 6) δ 12.62 (s, 1H), 8.91-8.64 (m, 2H), 8.52 (d, J = 4.5 Hz, 1H), .06 (m, 1H), 7.89-7.79 (m, 1H), 7.57-7.53 (m, 1H), 7.43-7.40 (m, 1H), 7.22 (d, J = 8.5 Hz, 1H), 6.59 (s, 1H), 5.06-4.96 (m, 1H), 4.5-4.40 (m, 2H), 4.26-4.11 (m, 3H), 3.88- 3.85 (m, 1H), 3.62 (s, 3H), 2.62-2.54 (m, 2H), 2.50-2.48 (m, 2H), 1.81-1.79 (m, 2H), 1.71- 25 1.67 (m, 2H).
Example 218a 5-Bromo(1,5-dimethyl-1H-pyrazolylamino) methylpyridin-2(1H)-one 218a 307 A solution of 5-bromomethyl(5-methyl-1H-pyrazolylamino)pyridin-2(1H)- one (2.8 g, 9.9 mmol) in anhydrous DMF (10 mL) was treated with 60% dispersion of NaH in mineral oil (0.51 g, 13 mmol) while stirring under nitrogen. After effervescence the reaction was stirred for an onal 30 s. At this time the reaction was treated with 5 iodomethane (0.98 g, 7.0 mmol) with continued stirring under nitrogen for 2 hours. Water (50 mL) was added slowly and the mixture was filtered. The filtrate was extracted with ethyl acetate (3 X 30 mL). The combined extract was concentrated under d pressure and the residue was purified by flush column chromatography eluting with 3:1 petroleum ether/ethyl acetate to afford 218a (0.70 g, 24%). MS: [M+H]+ 297. 10 Example 218b (4-{5-[(1,5-Dimethyl-1H-pyrazolyl)amino]methyloxo- 1,6-dihydro-pyridinyl}{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- dienyl}pyridinyl)methyl Acetate 218b A 50-mL round-bottomed flask equipped with a reflux condenser was charged with 218a (130 mg, 0.44 mmol), (3-(acetoxymethyl)(7,7-dimethyloxo-3,4,7,8-tetrahydro- 15 1H-cyclo-penta[4,5]pyrrolo[1,2-a]pyrazin-2(6H)-yl)pyridinyl)boronic acid 199e (175 mg, 0.44 mmol), PdCl2(dppf) (36 mg, 0.044 mmol), K3PO4 (343 mg, 1.32 mmol), sodium acetate (108 mg, 1.32 mmol), acetonitrile (10 mL), and water (0.5 mL). After three cycles of /argon flush, the mixture was heated at 100oC for 3 h. It was then filtered and the filtrate was evaporated in vacuo. The residue was ed by silica-gel column 20 chromatography eluting with 1:20 methanol/dichloromethane to afford 218b as a red solid (103 mg, 42%). MS-ESI: [M+H]+ 570.2 Example 218 3-[4-[5-[(1,5-dimethylpyrazolyl)amino]methyloxopyridyl]- 3-(hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- b]pyrazinone 218 25 A e of 218b (103 mg, 0.17 mmol), lithium hydroxide (42 mg, 1.75 mmol), THF (3 mL), i-propanol (2 mL), and water (1 mL) was stirred at room temperature for 0.5 h. The reaction mixture was concentrated under reduced pressure and d with water (4 mL). It was then extracted with dichloromethane (10 mL X 2) and the combined dichloromethane extract was concentrated under reduced pressure. The residue was purified with reverse-phase 30 PLC to afford 218 (29 mg, 48%) as white solid. MS-ESI: [M+H]+ 528.4. 1H NMR (500 MHz, DMSO-d6) δ 8.48 (d, J = 5.0 Hz, 1 H), 8.04 (s, 1 H), 8.02 (d, J = 2.5 Hz, 1 H), 7.38 (d, J = 2.0 Hz, 1 H), 7.32 (d, J = 5.0 Hz, 1 H), 6.55 (s, 1 H), 5.89 (s, 1 H), 4.97 (s, 1 H), 4.48-4.39 (m, 2 H), 4.24-4.16 (m, 3 H), 3.86-3.84 (m, 1 H), 3.58 (s, 3 H), 3.57 (s, 3 H), 2.62- 2.56 (m, 2 H), 2.42 (s, 2 H), 2.16 (s, 3 H), 1.22 (s, 6 H). 308 Example 219a 3-(3-Aminophenylamino)bromomethylpyrazin-2(1H)-one 219a OAc N OH N B OH O N H2N H2N NH NH O 199e OAc O N N N N N N Br Pd(dppf)Cl2, K3PO4, KOAc, CH3CN/H2O, O N 100 oC, 2 h 219a 219b To a solution of 3,5-dibromomethylpyrazin-2(1H)-one (536 mg, 2.0 mmol) and 5 benzene-1,3-diamine (324 mg, 3.0 mmol) in panol (18 mL) was added triethylamine (2.8 mL). The reaction e was stirred at 80oC overnight. Then the mixture was evaporated under reduced pressure to afford 219a (480 mg, 81%) as a white solid. MS-ESI: [M+H]+ 295.0 Example 219b (4-{6-[(3-Aminophenyl)amino]methyloxo-4,5- 10 dihydropyrazinyl}{4,4 -dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- dienyl}pyridinyl)methyl Acetate 219b A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 219a (480 mg, 1.62 mmol), (3-(acetoxymethyl)(7,7- dimethyloxo-3,4,7,8-tetrahydro-1H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazin-2(6H)- 15 yl)pyridinyl)boronic acid 199e (1.61 g, 4.05 mmol), f)Cl2 (134 mg, 0.162 mmol), potassium acetate (318 mg, 3.24 mmol), K3PO4 (706 mg, 3.24 mmol), acetonitrile (20 mL), and water (8 drops). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 2h. It was then filtered and the filtrate was evaporated in vacuo. The residue was ed by silica-gel column chromatography eluting with 20:1 ethyl acetate/methanol to 20 afford 219b (354 mg, 38%) as a yellow solid. MS-ESI: [M+H]+ 568.3 Example 219 3-[4-[6-(3-aminoanilino)methyloxo-pyrazinyl] (hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- b]pyrazinone 219 A e of 219b (283.5 mg, 0.50 mmol) and m hydroxide monohydrate (630 25 mg, 15.0 mmol) in i-propanol/THF (1:1, 8 mL) and water (2 mL) was stirred at 35oC for 0.5 h.
The mixture was evaporated in vacuo and the residue was diluted with water (3 mL). It was then extracted with dichloromethane (3 X 20 mL). The ed dichloromethane extract 309 was concentrated under reduced pressure and the residue was purified by reverse-phase prep- HPLC to afford 219 (170 mg, 79%) as a pale yellow solid. MS-ESI: [M+H]+ 526.4. 1H NMR (500 MHz, DMSO-d6) δ 8.85 (s, 1H), 8.47 (d, J = 5.5 Hz, 1H), 7.63 (s, 1H), 7.56 (s, 1H), 7.51 (d, J = 5.0 Hz, 1H), 6.92-6.91 (m, 2H), 6.57(s, 1H), 6.24-6.22 (m, 1H), 5.13 (s, 2H), 5 4.84-4.75 (m, 2H), 4.49-4.46 (m, 1H), 4.30-4.26 (m, 1H), 4.20-4.19 (m, 2H), 3.95-3.92 (m, 1H), 3.56 (s, 3H), 2.62-2.54 (m, 2H), 2.43 (s, 2H), 1.23 (s, 6H).
Example 220a (S)Chloro(1-methyl(5-(2-methyl(oxetan yl)piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)nicotinaldehyde 220a 10 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with (S)methyl(5-(2-methyl(oxetanyl)piperazin yl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 191j (1.5 g, 1.0 eq., 3.11 mmol), 4-bromochloronicotinaldehyde 104a (1.02 g, 1.5 eq., 4.67 mmol), PdCl2(dppf) (130 mg, 0.05 eq., 0.16 mmol), K3PO4 (1.32 g, 2 eq., 6.22 mmol), 15 sodium acetate (510 mg, 2.0 eq., 6.22 mmol), acetonitrile (35 mL), and water (1.0 mL). After three cycles of vacuum/argon flush, the mixture was heated at 90oC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under d pressure and the ing e was purified by silica-gel column chromatography eluting with 50:1 310 dichloromethane/ethanol to afford the 220a (1.1 g, 71%) as yellow solid. : [M+H]+ 495.3.
Example 220b (S)(3,4,6,7,8,9-Hexahydropyrazino[1,2-a]indol-2(1H)-yl) (1-methyl(5- (2-methyl(oxetanyl)piperazinyl)pyridinylamino)oxo-1,6- 5 opyridinyl)nicotinaldehyde 220b A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 220a (300 mg, 1.0 eq., 0.61 mmol), 1,2,3,4,6,7,8,9- octahydropyrazino[1,2-a]indole (128 mg, 1.2 eq., 0.73 mmol), Pd2(dba)3 (55 mg, 0.1 eq., 0.060 mmol), X-Phos (30 mg, 0.1 eq., 0.060 mmol), Cs2CO3 (390 mg, 2.0 eq., 1.22 mmol), 10 and dioxane (15.0 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100 oC for 16 h. It was then cooled to room temperature and ed. The filtrate was concentrated under d pressure and the resulting residue was purified by silica-gel column chromatography eluting with 40:1 dichloromethane/EtOH to afford 220b (100 mg, 26%) as yellow solid. MS-ESI: [M+H]+ 635.3. 15 Example 220 5-[2-(3,4,6,7,8,9-hexahydro-1H-pyrazino[1,2-a]indolyl) (hydroxymethyl)pyridyl]methyl[[5-[(2S)methyl(oxetanyl)piperazinyl] l]amino]pyridinone 220 A 50-mL single-neck round-bottomed flask was charged with 220b (100 mg, 1.0 eq., 0.15 mmol), NaBH4 (30 mg, 5.0 eq., 0.75 mmol), methanol (5 mL), and romethane (5 20 mL). The mixture was stirred at 00C for 10 min and quenched with water (5 mL). It was then filtered and the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase prep-HPLC to afford the title compound (10 mg, 10%). MS-ESI: [M+H]+ 637.5. 1H NMR (500 MHz, DMSO-d6) δ 8.61 (d, J = 2.0 Hz, 1H), 8.44 (s, 1H), 8.24 (d, J = 5.0 Hz, 1H), 7.82 (d, J = 3.0 Hz, 1H), 7.53 (d, J = 2.0 Hz, 1H), 7.39-7.36 (m, 1H), 7.24 (d, J 25 = 9.0 Hz, 1H), 6.99 (d, J = 5.0 Hz, 1H), 5.55 (s, 1H), 5.36-5.35 (m, 1H), .54 (m, 2H), 4.48-4.40 (m, 6H), 3.92-3.90 (m, 2H), 3.79-3.67 (m, 3H), 3.59 (s, 3H), 3.40-3.37 (m, 2H), 3.09-3.07 (m, 1H), 2.95-2.92 (m, 1H), 2.55-2.51 (m, 2H), 2.38-2.30 (m, 4H), 2.17-2.16 (m, 1H), 1.75-1.74 (m, 2H), 1.68-1.65 (m, 2H), 0.92 (d, J = 6.5 Hz, 3H).
Example 221a 3-[(6-aminopyridinyl)amino]bromomethyl-1,2- 30 dihydropyridinone 221a 311 OAc N OH N B OH O N H2N N NH H2N N NH OAc O O 199e N N N N Br Pd(dppf)Cl2, K3PO4, KOAc, O N CH3CN/H2O, 100 oC, 2 h 221a 221b A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 1,4-dioxane (20 mL), 3,5-dibromomethylpyridin- 2(1H)-one (1.06 g, 4.0 mmol), pyridine-2,6-diamine (872 mg, 8.0 mmol), Pd2(dba)3 (732 mg, 5 0.80 mmol), XantPhos (462.4 mg, 0.80 mmol), and cesium ate (2.6 g, 8.0 mmol). After three cycles of vacuum/argon flush, the mixture was heated at 110oC for 1 h. After this time the reaction was cooled to room temperature. It was then filtered and the te was evaporated in vacuo. The residue was purified by -gel column chromatography eluting with 20:1 ethyl e/methanol to afford 221a (570 mg, 48%) as a white solid. MS-ESI: 10 [M+H]+ 295.0 Example 221b (4-{5-[(6-Aminopyridinyl)amino]methyloxo-1,6- dihydropyridinyl}{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- 0-yl}pyridinyl)methyl Acetate 221b A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 15 reflux condenser was charged with 221a (354 mg, 1.2 mmol), (3-(acetoxymethyl)(7,7- dimethyloxo-3,4,7,8-tetrahydro-1H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazin-2(6H)- yl)pyridinyl)boronic acid 199e (1.20 g, 3.0 mmol), Pd(dppf)Cl2 (99 mg, 0.12 mmol), potassium acetate (235 mg, 2.4 mmol), K3PO4 (532 mg, 2.4 mmol), acetonitrile (12 mL), and water ( 10 drops). After three cycles of vacuum/argon flush, the mixture was heated at 100oC 20 for 2 h. It was then filtered and the te was evaporated in vacuo. The residue was purified by silica-gel column chromatography eluting with 30:1 ethyl acetate/methanol to afford 221b (210 mg, 31%) as a yellow solid. MS-ESI: [M+H]+ 568.3 Example 221 3-[4-[5-[(6-aminopyridyl)amino]methyloxopyridyl] (hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- 25 b]pyrazinone 221 A mixture of 221b (181 mg, 0.32 mmol) and lithium ide monohydrate (148 mg, 3.2 mmol) in i-propanol/THF (1:1, 6 mL) and water (1.5 mL) was stirred at 35oC for 0.5 h.
The mixture was concentrated under reduced pressure and the residue was purified by 312 reverse-phase prep-HPLC to afford 221 (82 mg, 49%) as a pale yellow solid. MS-ESI: [M+H]+ 526.3. 1H NMR (500 MHz, DMSO-d6) δ 8.84 (d, J = 2.0 Hz, 1H), 8.46 (d, J = 5.0 Hz, 1H), 8.11 (s, 1H), 7.54 (d, J = 2.0 Hz, 1H), 7.38 (d, J = 5.0 Hz, 1H), 7.23 (d, J = 8.0 Hz, 1H), 6.57(s, 1H), 6.36 (d, J = 7.5 Hz, 1H), 5.91 (d, J = 8.0 Hz, 1H), 5.79 (bs, 2H), 5.07 (t, J 5 = 5.0 Hz, 1H), .47 (m, 2H), 4.27-4.20 (m, 3H), 3.90 (d, J = 10.5 Hz, 1H), 3.60 (s, 3H), 2.62-2.57 (m, 2H), 2.43 (s, 2H), 1.22 (s, 6H).
Example 222a N-(5-Chloromethoxypyridinyl)methylpyrimidin amine 222a N N N HN N NH OAc O O 199e N N N N Cl O N Pd2(dba)3, Cy3P, 222a Cs2CO3, dioxane 222b H2O, sealed, 110oC, 4 h N N LiOH, iPrOH,THF, HN H2O, OH O N 35oC, 0.5 h N N O N 222c 10 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with 1,4-dioxane (30 mL), 3-bromochloro ypyridine (865 mg, 3.9 mmol), 2-methylpyrimidinamine (327 mg, 3.0 mmol), Pd2(dba)3 (275 mg, 0.30 mmol), XantPhos (173.4 mg, 0.30 mmol), and cesium ate (1.96 g, 6.0 mmol). After three cycles of vacuum/argon flush, the mixture was heated at 15 100oC for 5 h. After this time the reaction was cooled to room temperature. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified by silica-gel column chromatography eluting with 5:1 ethyl acetate/petroleum ether to afford 222a (555 mg, 74%) as a white solid. MS-ESI: [M+H]+ 251.0 313 Example 222b (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl} {6-methoxy[(2-methylpyrimidinyl)amino]pyridinyl}pyridin yl)methyl Acetate 222b A sealed tube ed with a magnetic stirrer was d with 222a (550 mg, 2.2 5 mmol), (3-(acetoxymethyl)(7,7-dimethyloxo-3,4,7,8-tetrahydro-1H- cyclopenta[4,5]pyrrolo-[1,2-a]pyrazin-2(6H)-yl)pyridinyl)boronic acid 199e (2.18 g, 5.5 mmol), Pd2(dba)3 (201 mg, 0.22 mmol), tricyclohexylphospine (84 mg, 0.30 mmol), Cs2CO3 (1.43 g, 4.4 mmol), dioxane (12 mL), and water (1 mL). After three cycles of vacuum/argon flush, the mixture was heated at 110oC for 4 h. It was then filtered and the filtrate was 10 evaporated in vacuo. The residue was ed by silica-gel column chromatography eluting with 30:1 ethyl acetate/methanol to afford 222b (310 mg, 25%) as a yellow solid. MS-ESI: [M+H]+ 568.6 e 222c 10-[3-(Hydroxymethyl){6-methoxy[(2-methylpyrimidin- 4-yl)amino]pyridinyl}pyridinyl]-4,4-dimethyl-1,10-diazatricyclo[6.4.0.02,6]dodeca- 15 2(6),7- dienone 222c A mixture 222b (283.5 mg, 0.50 mmol) and lithium hydroxide monohydrate (630 mg, 15.0 mmol) in anol/THF (1:1, 10 mL) and water (2.5 mL) was stirred at 35 oC for 0.5 h.
The mixture was ated in vacuo and the residue was diluted with water (3 mL). It was then ted with dichloromethane (3 X 20 mL). The ed organic extract was 20 concentrated under reduced pressure to afford 222c (240 mg, 92%) as a white solid. MS-ESI: [M+H]+ 526.2 Example 222 3-[3-(hydroxymethyl)[5-[(2-methylpyrimidinyl)amino]oxo- 1H-pyridinyl]pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- b]pyrazinone 222 25 To a solution of 222c (226 mg, 0.43 mmol) in dioxane (8 mL) was added concentrated HCl (1.1 mL). The reaction was stirred at 100 oC for 1 h. Then the mixture was adjusted to pH 7.0 by introducing saturated aqueous NaHCO3. It was extracted with dichloromethane (3 x 20 mL) and the combined extract was evaporated under reduced pressure. The resulting residue was purified by reverse-phase prep-HPLC to afford 222 (30 30 mg, 14%) as a white solid. MS-ESI: [M+H]+ 512.3. 1H NMR (500 MHz, DMSO-d6) δ 12.24 (s, 1H), 9.10 (s, 1H), 8.96 (d, J = 2.0 Hz, 1H), 8.50 (d, J = 5.0 Hz, 1H), 8.22 (d, J = 6.5 Hz, 1H), 7.50 (d, J = 2.0 Hz, 1H), 7.40 (d, J = 5.0 Hz, 1H), 7.13 (d, J = 6.0 Hz, 1H), 6.57 (s, 1H), 5.08-5.06 (m, 1H), 4.50-4.42 (m, 2H), 4.25-4.19 (m, 3H), 3.87-3.85 (m, 1H), 2.62-2.53 (m, 2H), 2.45 (s, 3H), 2.43 (s, 2H), 1.23 (s, 3H), 1.22 (s, 3H) 314 Example 223a 2-(10-Fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- yl)(1-methyl(5-methyl-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridinylamino)oxo-1,6- dihydropyridinyl)nicotinaldehyde 223a 5 A 50-mL round-bottomed flush equipped with a reflux ser was charged with 1- methyl(5-methyl-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridinylamino)(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolanyl)pyridin-2(1H)-one 216a (200 mg, 0.50 mmol), 4-chloro (10-fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 134c (174 mg, 0.50 mmol), K3PO4 (212 mg, 1.0 mmol), sodium acetate (82mg, 1.0 mmol), 1,1’- 10 bis(diphenylphosphino)ferrocenedichloropalladium(II) (21 mg, 0.025 mmol), and acetonitrile/water(15/1 mL). After bubbling nitrogen h the mixture for 30 minutes, it was heated at 100°C for 1h under N2 protection. Analysis of reaction mixture by LCMS showed te conversion to the desired product. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue 15 was diluted with dichloromethane (50 mL) and water (50 mL). The s layer was extracted with romethane (3 X 20 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The dark residue was ed by silica-gel column chromatography eluting with dichloromethane/methanol (70/1 to 30/1) to afford 223a (167 mg, 57%) as yellow solid. : [M+H]+ 588.1 20 Example 223 10-fluoro[3-(hydroxymethyl)[1-methyl[(5-methyl-6,7- dihydro-4H-thiazolo[5,4-c]pyridinyl)amino]oxopyridyl]pyridyl]-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indolone 223 Compound 223a (160 mg, 0.27 mmol) was dissolved in methanol (30 mL), followed by addition of NaBH4 (31 mg, 082 mmol) at 0°C. The reaction e was stirred for 30 min 25 and then quenched with water (10 mL). It was concentrated under reduced pressure and the residue was extracted with dichloromethane (3 X 30 mL). The combined organic phase was concentrated under reduced pressure and the residual was purified by reverse-phase prep- 315 HPLC to afford 223 (56 mg, 35%) as a white solid. MS-ESI: [M+H]+ 590.2. 1H NMR (500 MHz, DMSO-d6): δ 9.92 (s,1H), 8.64 (d, J = 2.5 Hz,1H), 8.48 (d, J = 5.5 Hz, 1H), 7.56 (d, J = 2.0 Hz, 1H), 7.34 (d, J = 5.0 Hz, 1H), 4.93 (t, J = 5.5 Hz, 1H), .41 (m, 2H), 4.19-4.17 (m, 2H), 4.08-4.03 (m, 1H), 3.88-3.85 (m, 1H ), 3.60 (s, 3H), 3.42 (s, 2H), 2.63-2.58 (m, 6H), 5 2.41(s, 3H), 2.34 (s, 2H), 1.78-1.76 (m, 2H), 1.68-1.66 (m, 2H).
Example 224a (S)-tert-Butyl 4-(6-(6-Chloromethyloxo-2,3- dihydropyridazinylamino)pyridinyl)methylpiperazinecarboxylate 224a O Boc N HN N N N N O N NH N NH N NH O HCl/EtOH O O O N 25 ° C, 2 h N ZnCl2, Cl N Cl N N NaBH3CN Cl N MeOH, 224a 224b 50 °C, 3h 224c O N N N NH 199e OAc O N Pd(dppf)Cl2, K3PO4, N N N NaOAc,CH3CN/H2O, 100 °C, 1 h, O N 224d A 250-mL single-neck round-bottomed flask equipped with a ic stirrer and a 10 reflux condenser was d with (S)-tert-butyl 4-(6-aminopyridinyl) methylpiperazinecarboxylate 191f (2.5 g, 8.5 mmol), 4-bromochloro pyridazin-3(2H)-one (2.2 g, 10.0 mmol), XantPhos (240 mg, 0.40 mmol), tris(dibenzylideneacetone)dipalladium(0) (360 mg, 0.40 mmol), Cs2CO3 (5.5 g, 17 mmol), and 1,4-dioxane (100 mL). After three cycles of vacuum/argon flush, the mixture was heated 15 at 100oC for 2.5 h. It was then cooled to room ature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (40:1 to 30:1) to afford 224a as a pale yellow solid (3.2 g, 86%). MS-ESI: [M+H]+ 435.1. 316 Example 224b (S)Chloromethyl(5-(2-methylpiperazinyl)pyridin ylamino)pyridazin-3(2H)-one 224b A mixture of 224a (3.0 g, 6.9 mmol) and 4.0M hanol (20 mL) was stirred at room temperature for 2 h. The mixture was then trated under reduced pressure to 5 afford crude 224b as a yellow solid (2.5 g, 98%), which was used for the next step without further purification. MS-ESI: [M+H]+ 335.1.
Example 224c (S)Chloromethyl(5-(2-methyl(oxetan yl)piperazinyl)pyridinylamino)pyridazin-3(2H)-one 224c A mixture of 224b (2.3 g, 6.8 mmol), oxetanone (1.4 g, 20.0 mmol), NaBH3CN 10 (620 mg, 10 mmol), and zinc chloride (1.36 g, 10 mmol) in methanol (20 mL) was stirred at 50oC for 3 hours. The mixture was added to water (40 mL) and concentrated under reduced pressure. The residue was extracted with romethane three times. The combined organic layer was dried and concentrated under reduced pressure. The residue was purified by gel column chromatography eluting with 50:1 dichloromethane/methanol to afford 224c (2.0 15 g, 75%). MS-ESI: [M+H]+ 391.2.
Example 224d (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}[1-methyl({5-[(2S)methyl(oxetanyl)piperazinyl]pyridin- 2-yl}amino)oxo-1,6-dihydropyridazinyl]pyridinyl)methyl Acetate 224d A sealed tube equipped with a magnetic stirrer was charged with 224c (200 mg, 0.50 20 mmol), {3-[(acetoxy)methyl]{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}pyridinyl}boronic acid 199e (240 mg, 0.60 mmol), Pd(dppf)Cl2 (18 mg, 0.025 mmol), sodium acetate (74 mg, 0.90 mmol), K3PO4 (191 mg, 0.90 mmol), and acetonitrile/water (6:1, 3.5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 1 h. It was then cooled to room temperature and ed. The te was 25 evaporated under reduced pressure and the residue was purified by silica-gel column tography eluting with 25:1 dichloromethane/methanol to afford 224d (180 mg, 51%) as a brown solid. MS-ESI: [M+H]+ 708.3.
Example 224 3-[3-(hydroxymethyl)[1-methyl[[5-[(2S)methyl(oxetan yl)piperazinyl]pyridyl]amino]oxo-pyridazinyl]pyridyl]-7,7-dimethyl-1,2,6,8- 30 tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 224 A mixture of 224d (180 mg, 0.25 mmol) and lithium hydroxide (72 mg, 3.0 mmol) in i-propanol/THF (5/3 mL) and water (2 mL) was stirred at 35 oC for 0.2 h. The e was evaporated under reduced pressure and the residue was extracted with ethyl acetate (10 mL X 2). The combined ethyl acetate extract was concentrated under reduced pressure and the 317 residue was purified by reverse phase lush eluting with 0.3% NH4HCO3 in water/acetonitrile to afford 224 (54 mg, 33%) as a white solid. : [M+H]+ 666.3. 1H NMR (500 MHz, DMSO-d6) δ 9.29 (s, 1H), 8.52 (d, J = 4.0 Hz, 1H), 8.43 (s, 1H), 7.91 (s, 1H), 7.43-7.42 (m, 2H), 7.40-7.39 (m, 1H), 6.55 (s, 1H), 4.77-4.75 (m, 1H), 4.57-4.55 (m, 5 3H), 4.48-4.47 (m, 1H), .41 (m, 2H), 4.28-4.26 (m, 1H), .18 (m, 2H), 3.88-3.86 (m, 2H), 3.77 (s, 3H), 3.38-3.37 (m, 1H), 3.21-3.19 (m, 1H), 2.98-2.96 (m, 1H), 2.64-2.62 (m, 1H), 2.58-2.56 (m, 2H), 2.42-2.41 (m, 3H), .25 (m, 1H), 2.11-2.09 (m, 1H), 1.21 (s, 6H), 0.98 (d, J = 5.5 Hz, 3H).
Example 225a romethyl(2-methylpyrimidinylamino)pyridazin- 10 3(2H)-one 225a A 100-mL single-neck round-bottomed flask equipped with a reflux condenser was charged with 2-methylpyrimidinamine (330 mg, 3.03 mmol), 4-bromochloro methylpyridazin-3(2H)-one (675 mg, 3.03 mmol), Pd2(dba)3 (274 mg, 0.30 mmol), XantPhos 15 (143 mg, 0.30 mmol), Cs2CO3 (2960 mg, 9.09 mmol), and dioxane (40 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for overnight. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified by silica-gel column chromatography eluting with 1:20 methanol/dichloromethane to afford 225a as a yellow solid (560 mg, 73%). MS-ESI: [M+H]+ 252.1 20 Example 225b (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{1-methyl[(2-methylpyrimidinyl)amino]oxo-1,6- dihydropyridazinyl}pyridinyl)methyl Acetate 225b A round-bottomed flask equipped with a reflux condenser was charged with 225a (200 mg, 0.80 mmol), (3-(acetoxymethyl)(7,7-dimethyloxo-3,4,7,8-tetrahydro-1H- 25 cyclopenta[4,5]pyrrolo[1,2-a]pyrazin-2(6H)-yl)pyridinyl)boronic acid 199e (318 mg, 0.80 mmol), PdCl2(dppf) (65.3 mg, 0.080 mmol), K3PO4 (624 mg, 2.4 mmol), sodium acetate (200 mg, 2.4 mmol), acetonitrile (10 mL), and water (0.5 mL). After three cycles of vacuum/argon flush, the e was heated at 100 oC for 3 h. It was then filtered and the filtrate was 318 evaporated in vacuo. The e was purified by silica-gel column chromatography eluting with 1:20 methanol/dichloromethane to afford 225b as a red solid (150 mg, 47%). MS-ESI: [M+H]+ 569.3 Example 225 3-[3-(hydroxymethyl)[1-methyl[(2-methylpyrimidin 5 yl)amino]oxo-pyridazinyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 225 A mixture of 225b (120 mg, 0.21 mmol), lithium ide (59 mg, 2.11 mmol), THF (6 mL), i-propanol (4 mL), and water (2 mL) was stirred at room ature for 0.5 h. The mixture was concentrated under reduced pressure and diluted with water (3 mL). It was then 10 extracted with dichloromethane (2 X 10 mL). The combined dichloromethane extract was concentrated under reduced re and the residue was ed with reverse-phase prep- HPLC to afford 225 as a white solid (29 mg, 48%). MS-ESI: [M+H]+ 527.3. 1H NMR (500 MHz, DMSO-d6) δ 9.82 (s, 1 H), 8.88 (s, 1 H), 8.54 (d, J = 5.0 Hz, 1 H), 8.37 (d, J = 6.0 Hz, 1 H), 7.44 (d, J = 4.5 Hz, 1 H), 7.34 (d, J = 3.5 Hz, 1 H), 6.56 (d, J = 4.0 Hz, 1 H), 4.87 (t, J 15 = 1.5 Hz, 1 H), 4.67 (d, J = 11.5 Hz, 1 H), 4.42 (d, J = 12.5 Hz, 1 H), 4.29-4.25 (m, 1 H), 4.20 (bs, 2 H), 3.93 (d, J = 9.5 Hz, 1 H), 3.81 (s, 3 H), 2.62-2.58 (m, 2 H), 2.50-2.49 (m, underneath solvent peak, 2H), 2.40 (s, 3 H), 1.22 (s, 6 H).
Example 226a 6-Chloromethyl({5-[(morpholinyl)carbonyl]pyridin yl}amino)-2,3-dihydropyridazinone 226a 20 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with 1,4-dioxane (40 mL), (6-aminopyridin yl)(morpholino)methanone 111a (2.07 g, 10.0 mmol), 4-bromochloromethylpyridazin- 3(2H)-one (3.35 g, 15.0 mmol), a)3 (915 mg, 1.0 mmol), XantPhos (578 mg, 1.0 25 mmol), and cesium carbonate (6.52 g, 20 mmol). After three cycles of vacuum/argon flush, the mixture was heated at 100 oC for 8 h. It was then cooled to room temperature and filtered.
The solid was washed with dichloromethane (2 X 20 mL). The combined filtrate was dried over anhydrous Na2SO4 and concentrated under reduced pressure to afford 226a (2.45 g, 51%) as a yellow solid. MS: [M+H]+ 350.1 319 Example 226b (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- -dienyl} [1-methyl({5-[(morpholinyl)carbonyl]pyridinyl}amino) oxo-1,6-dihydro- pyridazinyl]pyridinyl)methyl Acetate 226b A sealed tube equipped with a ic stirrer was charged with 226a (279 mg, 0.80 5 mmol), (2-{4,4-dimethyloxo-1,10-diazatricyclo [6.4.0.02,6]dodeca-2(6),7-dienyl} (tetramethyl-1,3,2-dioxaborolanyl)pyridinyl) methyl acetate 199e (1.53 g, 3.2 mmol), Pd2(dba)3 (73.2 mg, 0.080 mmol), tricyclohexyl-phospine (44.6 mg, 0.16 mmol), cesium carbonate (521.6 mg, 1.6 mmol), 1,4-dioxane (10 mL), and water ( 8 drops). After three cycles of vacuum/argon flush, the mixture was stirred at 110oC for 4 h. It was then filtered 10 and the filtrate was evaporated in vacuo. The e was purified by silica-gel column tography eluting with 40:1 ethyl acetate/methanol to afford 226b as a yellow solid (120 mg, 23%). MS-ESI: [M+H]+ 667.3 Example 226 3-[3-(hydroxymethyl)[1-methyl[[5-(morpholinecarbonyl) pyridyl]amino]oxo-pyridazinyl]pyridyl]-7,7-dimethyl-1,2,6,8- 15 tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 226 A mixture of 226b (120 mg, 0.18 mmol) and m hydroxide monohydrate (227 mg, 5.4 mmol) in i-propanol/THF/water (3 mL /3 mL /2 mL) was stirred at 35oC for 0.5 h. The mixture was evaporated in vacuo and the residue was extracted with dichloromethane (3 X 10 mL). The combined dichloromethane t was concentrated under reduced pressure and 20 the e was purified by reverse-phase prep-HPLC to afford 226 as a white solid (53 mg, 47%). MS-ESI: [M+H]+ 625.3. 1H NMR (500 MHz, DMSO-d6) δ 9.75 (s, 1H), 8.64 (s, 1H), 8.54 (d, J = 5.0 Hz, 1H), 8.36 (d, J = 2.5 Hz, 1H), 7.79-7.77 (m, 1H), 7.59 (d, J = 8.5 Hz, 1H), 7.43 (d, J = 5.0 Hz, 1H), 6.56 (s, 1H), 4.82 (s, 1H), 4.60-4.57 (m, 1H), 4.40-4.37 (m, 1H), 4.27-4.25 (m, 1H), 4.20-4.17 (m, 2H), 3.91-3.88 (m, 1H), 3.80 (s, 3H), 3.60-3.45 (m, 25 overlap, 8H), 2.62-2.56 (m, 2H), 2.42 (s, 2H), 1.22 (s, 6H).
Example 227a {4-[1-Methyl({5-[(morpholinyl)carbonyl]pyridin yl}amino)oxo-1,6-dihydropyridazinyl]{1-oxo-1H,2H,3H,4H,6H,7H,8H,9H- pyrazino[1,2-a]indolyl}pyridinyl}methyl Acetate 227a 320 A sealed tube equipped with a magnetic stirrer was charged with 6-chloromethyl- 4-({5-[(morpholinyl)carbonyl]pyridinyl}amino)-2,3-dihydropyridazinone 226a (244 mg, 0.70 mmol), 3-(acetoxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 5 2(1H)-yl)pyridinylboronic acid 113i (558 mg, 1.5 mmol), Pd2(dba)3 (64 mg, 0.070 mmol), tricyclohexylphospine (39 mg, 0.14 mmol), cesium carbonate (456 mg, 1.4 mmol), 1,4- e (7 mL), and water ( 6 drops). After three cycles of vacuum/argon flush, the mixture was stirred at 110oC for 4 h. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified by silica-gel column tography eluting with 40:1 ethyl 10 acetate/methanol to afford 227a as a yellow solid (290 mg, 63%). : [M+H]+ 653.3 Example 227 2-[3-(hydroxymethyl)[1-methyl[[5-(morpholinecarbonyl) pyridyl]amino]oxo-pyridazinyl]pyridyl]-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 1-one 227 A mixture of 227a (131 mg, 0.20 mmol) and lithium hydroxide·1water (120 mg, 2.0 15 mmol) in i-propanol/THF/water (4 mL /4 mL /2 mL) was stirred at 35oC for 0.5 h. The mixture was evaporated in vacuo and the residue was extracted with dichloromethane (3 X 10 mL). The combined dichloromethane extract was concentrated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 227 as a white solid (75 mg, 62%). MS-ESI: [M+H]+ 611.2. 1H NMR (500 MHz, DMSO-d6) δ 9.75 (s, 1H), 8.64 (s, 1H), 20 8.54 (d, J = 5.0 Hz, 1H), 8.36 (d, J = 2.0 Hz, 1H), 7.79-7.77 (m, 1H), 7.59 (d, J = 8.5 Hz, 1H), 7.43 (d, J = 5.0 Hz, 1H), 6.58 (s, 1H), 4.82 (s, 1H), 4.60-4.57 (m, 1H), 4.38-4.36 (m, 1H), .19 (m, 2H), 4.10-4.05 (m, 1H), 3.93-3.90 (m, 1H), 3.80 (s, 3H), 3.60-3.50 (m, overlap, 8H), 2.66-2.54 (m, 2H), .46 (m, 2H), .66 (m, 4H).
Example 228a (S)-(4-(1-Methyl(5-(2-methyl(oxetanyl)piperazin 25 yl)pyrazinyl-amino)oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl Acetate 228a 321 O N N N N NH AcO O N N N O N 228a A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with (S)bromomethyl(5-(2-methyl(oxetan yl)piperazinyl)pyrazinylamino)pyridin-2(1H)-one (90 mg, 0.21 mmol) 191i, 3- 5 (acetoxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridin ylboronic acid 113i (80.4 mg, 0.21 mmol), Pd(dppf)Cl2 (17.2 mg, 0.021 mmol), K3PO4 (89 mg, 0.42 mmol), sodium acetate (57.1 mg, 0.42 mmol), water (0.5 mL), and acetonitrile (30 mL). After three cycles of vacuum/argon flush, the mixture was heated at reflux for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under 10 reduced re and the ing residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (50:1 to 30:1) to afford 228a as brown solid (60 mg, 42%). MS-ESI: [M+H]+ 694.3.
Example 228 2-[3-(hydroxymethyl)[1-methyl[[5-[(2S)methyl(oxetan yl)piperazinyl]pyrazinyl]amino]oxopyridyl]pyridyl]-3,4,6,7,8,9- 15 hexahydropyrazino[1,2-a]indolone 228 A e of 228a (50 mg, 0.070 mmol) and lithium hydroxide (43 mg, 1.8 mmol) in i-propanol /THF (1:1, 4 mL) and water (1 mL) was stirred at 35oC for 30 mins. The reaction e was trated under reduced pressure and the residue was purified by reversephase prep-HPLC to afford 228 (10 mg, 21%). MS-ESI: [M+H]+ 652.3. 1H NMR (500 MHz, 20 DMSO-d6) δ 8.61 (s, 1H), 8.48 (d, J = 4.5 Hz, 1H), 8.39 (s, 1H), 8.34 (s, 1H), 7.84 (s, 1H), 7.48 (s, 1H), 7.34 (d, J = 5.0 Hz, 1H), 6.58 (s, 1H), 4.94 (bs, 1H), 4.56-4.55 (m, 2H), 4.49- 4.47 (m,1H), 4.42-4.36 (m, overlap, 4H) 4.25-4.17 (m, 2H), 4.13-4.10 (m, 1H), 3.86-3.76 (m, 2H), 3.60 (s, 3H), 3.39-3.37 (m, 1H), 3.01-2.96 (m, 1H), 2.78-2.76 (m, 1H), .57(m, overlap, 3H), 2.50-2.47 (m, 2H), 2.10-2.07 (m, 1H), 1.94-1.90 (m, 1H), 1.80-1.78 (m, 2H), 25 1.70-1.69 (m, 2H), 1.12 (d, J = 6.5 Hz, 3H).
Example 229a 2-Ethylpyrimidinamine 229a 322 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 2-chloropyrimidinamine (2.60 g, 20.0 mmol), triethylborane (20.0 mL, 1.0 M in THF, 20.0 mmol), Pd(dppf)Cl2 (816 mg, 1.0 mmol), K3PO4 5 (13.0 g, 40.0 mmol), water (2 mL), and tetrahydrofuran (50 mL). After three cycles of vacuum/argon flush, the mixture was heated at reflux for 14 h. It was then cooled to room temperature and ed. The filtrate was concentrated under reduced pressure and the resulting residue was purified by -gel column tography eluting with 40:1 dichloromethane/methanol to afford 229a as yellow solid (600 mg, 24%). MS-ESI: [M+H]+ 10 124.3 Example 229b 5-Bromo(2-ethylpyrimidinylamino)methylpyridin- 2(1H)-one 229b A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was d with 229a (246 mg, 2.0 mmol), 3,5-dibromomethylpyridin- 15 2(1H)-one (534 mg, 2.0 mmol), a)3 (182 mg, 0.20 mmol), XantPhos (231 mg, 0.40 mmol), Cs2CO3 (1.30 g, 4.0 mmol), and 1,4-dioxane (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 4 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 40:1 20 dichloromethane/methanol to afford 229b as off-white solid (308 mg, 50%). MS-ESI: [M+H]+ 309.1 Example 229c ,7-Dimethyloxo-3,4,7,8-tetrahydro-1H-cyclopenta- [4,5]pyrrolo[1,2-a]pyrazin-2(6H)-yl)((2-ethylpyrimidinyl)amino)methyloxo-1,6- dihydro-[3,4'-bipyridin]-3'-yl)methyl Acetate 229c 25 A 100-mL bottomed flask equipped with a reflux condenser was charged with 229b (277 mg, 0.90 mmol), (3-(acetoxymethyl)(7,7-dimethyloxo-3,4,7,8-tetrahydro- 1H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazin-2(6H)-yl)pyridinyl)boronic acid 199e (358 mg, 0.90 mmol), Pd(dppf)Cl2 (74 mg, 0.090 mmol), K3PO4 (381 mg, 1.80 mmol), water (2 mL), and tetrahydrofuran (20 mL). After three cycles of vacuum/argon flush, the mixture was 30 heated at reflux for 3 h. It was then cooled to room temperature and filtered. The filtrate was 323 concentrated under reduced pressure and the ing residue was purified by silica-gel column chromatography eluting with 60:1 dichloromethane/ methanol to afford 229c as white solid (291 mg, 50%). MS-ESI: [M+H]+ 582.4 Example 229 3-[4-[5-[(2-ethylpyrimidinyl)amino]methyloxopyridyl] 5 (hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- b]pyrazinone 229 To a solution of 229c (291 mg, 0.45 mmol) in tetrahydrofuran (10 mL) and water (2 mL) was added lithium hydroxide (48 mg, 2.0 mmol). The reaction mixture was stirred at 25oC for 1 h and concentrated under reduced pressure. The residue was purified by reverse- 10 phase prep-HPLC to afford 229 (165 mg, 61%) as yellow solid. MS-ESI: [M+H]+ 540.3. 1H NMR (500 MHz, DMSO-d6) δ 9.12 (s, 1H), 8.96 (d, J = 2.5 Hz, 1H), 8.50 (d, J = 5.0 Hz, 1H), 8.25 (d, J = 5.5 Hz, 1H), 7.72 (d, J = 2.5 Hz, 1H), 7.38 (d, J = 5.5 Hz, 1H), 7.15 (d, J = 6.0 Hz, 1H), 6.56 (s, 1H), 5.00 (t, J = 5.5 Hz, 1H), 4.55-4.44 (m, 2H), 4.26-4.19 (m, p, 3H), 3.88-3.86 (m, 1H), 3.62(s, 3H), 2.74 (q, J = 7.5 Hz, 2H), .54 (m, 2H), 2.43 (s, 2H), 15 .20 (m, overlap, 9H).
Example 230a (S)-tert-Butyl 4-(5-Bromopyrazinyl)methylpiperazine carboxylate 230a A mixture of rt-butyl 3-methylpiperazinecarboxylate (6.0 g, 30 mmol) and 20 2,5-dibromopyrazine (14.1 g, 60 mmol) was stirred at 80oC for 15 h. It was then cooled to room temperature and purified by silica-gel column chromatography eluting with petroleum ether/ethyl acetate (10:1 to 2:1) to afford 230a as a yellow solid (1.14 g, 19%). MS: [M+H]+ 359.1. 324 Example 230b (S)-tert-Butyl 4-(5-(Diphenylmethyleneamino)pyrazinyl) methylpiperazinecarboxylate 230b A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 5 reflux condenser was charged with 230a (2.6 g, 7.3 mmol), diphenylmethanimine (1.3 g, 7.3 mmol), Pd2(dba)3 (669 mg, 0.73 mmol), (R)-(+)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (968 mg, 1.46 mmol), Cs2CO3 (4.7 g, 14.6 mmol), and 1,4-dioxane (40 mL). After three cycles of vacuum/argon flush, the mixture was heated at 96oC for 3 hrs. It was then cooled to room temperature and filtered. The filtrate was trated under reduced pressure and the 10 resulting residue was purified by -gel column chromatography eluting with petroleum ether/ethyl acetate (10:1 to 3:1) to afford 230b as red oil (3.3 g, 75 %). MS: [M+H]+ 458.3. e 230c (S)-tert-Butyl 4-(5-Aminopyrazinyl)methylpiperazine carboxylate 230c To a solution of 230b (3.3 g, 7.2 mmol) in methanol (25 mL) were added sodium 15 acetate (708 mg, 8.6 mmol) and hydroxylamine hloride (907 mg, 8.6 mmol). The reaction mixture was stirred for 0.5 h. It was then concentrated in vacuo. The residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (50:1 to 30:1) to afford 230c as yellow oil (1.35 g, 64%). MS: [M+H]+ 294.3.
Example 230d rt-Butyl 4-(5-(5-Bromomethyloxo-1,2- 20 dihydropyridinylamino)pyrazinyl)methylpiperazinecarboxylate 230d A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was d with 230c (1.25 g, 4.3 mmol), 3,5-dibromomethylpyridin- 2(1H)-one (3.4 g, 12.9 mmol), Pd2(dba)3 (394 mg, 0.43 mmol), Xantphos (497 mg, 0.86 mmol), Cs2CO3 (4.7 g, 14.6 mmol), and 1,4-dioxane (80 mL). After three cycles of 25 vacuum/argon flush, the mixture was heated at 90oC for 2 hrs. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was ed by silica-gel column chromatography eluting with dichloromethane/methanol (50:1 to 30:1) to afford 230d (1.9 g, 72 %). MS: [M+H]+ 481.2.
Example 230e (S)Bromomethyl(5-(2-methylpiperazinyl)pyrazin 30 ylamino)pyridin-2(1H)-one 230e A mixture of 230d (1.9 g, 3.97 mmol) and trifluoroacetic acid (4 mL) was stirred at room temperature for 1 h. It was then concentrated under reduced re to afford crude 230e (1.45 g, 97 %), which was used in the next step without further purification. MS: [M+H]+ 381.2. 325 Example 230f (S)Bromomethyl(5-(2-methyl(oxetan erazinyl)pyrazinylamino)pyridin-2(1H)-one 230f A mixture of 230e (2.0 g, 5.3 mmol), oxetanone (763 mg, 10.6 mmol), NaBH3CN (835 mg, 13.3 mmol), and zinc chloride (1.8 g, 13.3 mmol) in methanol (60 mL) was stirred 5 at 50oC for 30 min. The e was concentrated under reduced pressure. To the residue was added water and the resulting mixture was extracted with dichloromethane three times. The combined organic layer was then concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (50:1 to 20:1) to afford 230f as a yellow oil (1.6 g, 70%). MS: 10 [M+H]+ 437.2.
Example 230g 5-[4-Chloro(hydroxymethyl)pyridinyl]thia-4,5- ricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienone 230g A e of 4-chloro{6-oxothia-4,5- diazatricyclo[7.4.0.02,7]trideca- 15 1(9),2(7),3-trine yl}pyridine baldehyde 124a (797 mg, 2.31 mmol), NaBH4 (263 mg, 6.92 mmol), and CH3OH (50 mL) was stirred at room temperature for 1 h. Then the reaction mixture was quenched with water (30 mL) and concentrated under reduced pressure. The residue was extracted with dichloromethane (2 X 30 mL) and the combined dichloromethane extract was concentrated under d pressure. The residue was purified by silica-gel 20 chromatography eluting with 5:1 to afford 230g (649 mg, 81%) as a yellow solid. MS-ESI: [M+H]+ 348.1 Example 230h (4-Chloro{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca- 1(9),2(7),3-trien yl}pyridinyl)methyl Acetate 230h 326 A bottomed flask was d with 230g (597 mg, 1.72 mmol), dichloromethane (50 mL), and ylamine (5 mL). The solution was stirred at 0oC for 0.5 h and acetyl chloride (135 mg, 1.72 mmol) was added slowly. The mixture was stirred at 0oC for r 2.5 h. It was then concentrated under reduced pressure. The residue was purified 5 by silica-gel column chromatography eluting with 9:1 petroleum ether/ethyl acetate to afford 230h (602 mg, 90%) as a yellow solid. MS-ESI: [M+H]+ 390.1 Example 230i ydroxyboranyl){6-oxothia-4,5-diazatricyclo- [7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridinyl]methyl acetate 230i A 100-mL -neck round-bottomed flask equipped with a magnetic stirrer and a 10 reflux condenser was charged with 230h (595 mg, 1.53 mmol), Pin2B2 (1.94 g, 7.65 mmol), PdCl2(dppf) (65 mg, 0.080 mmol), X-Phos (73 mg, 0.15 mmol), potassium acetate (304 mg, 3.1 mmol), and dioxane (50 mL). After three cycles of vacuum/argon flush, the mixture was heated at 65ºC for 5 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was washed with petroleum 15 ether to afford 230i (409 mg, 67%) as a yellow solid. MS-ESI: [M+H]+ 400.1 Example 230j {4-[1-Methyl({5-[(2S)methyl(oxetanyl)piperazin azinyl}amino)oxo-1,6-dihydropyridinyl]{6-oxothia-4,5- diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridinyl}methyl Acetate 230j A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 20 reflux condenser was charged with 230f (100 mg, 0.23 mmol), 230i (140 mg, 0.35 mmol), Pd(dppf)Cl2 (19 mg, 0.023 mmol), sodium acetate (63 mg, 0.46 mmol), K3PO4 (98 mg, 0.46 mmol), water (0.5 mL), and acetonitrile (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at 90oC for 3 h. After cooling to room temperature the reaction was filtered. The filtrate was concentrated under reduced pressure and the resulting residue was 25 ed by silica-gel column chromatography g with dichloromethane/methanol (50:1 to 30:1) to afford 230j as a yellow solid (90 mg, 55%). MS-ESI: [M+H]+ 710.2.
Example 230 3-[3-(hydroxymethyl)[1-methyl[[5-[(2S)methyl(oxetan yl)piperazinyl]pyrazinyl]amino]oxopyridyl]pyridyl]-6,7,8,9- tetrahydrobenzothiopheno[2,3-d]pyridazinone 230 30 A mixture of 230j (80 mg, 0.11 mmol) and lithium hydroxide (27 mg, 1.1 mmol) in ipropanol /THF (1:1, 10 mL) and water (2 mL) was stirred at 35oC for 0.5 h. It was then cooled to room temperature and ed. The filtrate was concentrated under reduced pressure and the resulting residue was purified by reverse-phase prep-HPLC to afford 230 (34 mg, 45%). MS-ESI: [M+H]+ 668.2. 1H NMR (500 MHz, DMSO-d6) δ 8.64 (s, 1H), 8.56 (d, J = 327 6.0 Hz, 1H), 8.46 (s, 1H), 8.40 (d, J = 2.5 Hz, 1H), 8.34 (d, J = 1.5 Hz, 1H), 7.85 (s, 1H), 7.53 (d, J = 6.0 Hz, 1H), 7.49 (d, J = 2.5 Hz, 1H), 4.84 (bs, 1H), 4.58-4.54 (m, 2H), 4.50- 4.47 (m, 1H), 4.43-4.36 (m, overlap, 4H), 3.78-3.75 (m, 1H), 3.59 (s, 3H), 3.39-3.35 (m, 1H), 3.02-3.0 (m, 1H), 2.98-2.95 (m, 2H), 2.90-2.82 (m, 2H), 2.78-2.76 (m, 1H), 2.60-2.56 (m, 5 1H), 2.10-2.09 (m, 1H), 1.95-1.88 (m, overlap, 5H), 1.10 (d, J = 8.0 Hz, 3H).
Example 231a 2-(4-Chloro(hydroxymethyl)pyridinyl)fluoro- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-1(2H)-one 231a A mixture of 4-chloro(10-fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 10 2(1H)-yl)nicotinaldehyde 134c (800 mg, 2.31 mmol), NaBH4 (263 mg, 6.92 mmol), and methanol (50 mL) was stirred at 0oC for 1 h. Then the reaction mixture was quenched with water (30 mL) and concentrated under reduced pressure. The residue was extracted with dichloromethane (2 X 30 mL) and the combined dichloromethane extract was concentrated under reduced pressure. The residue was purified by -gel tography g with 15 5:1 petroleum ether/ethyl acetate to afford 231a (650 mg, 81%) as a yellow solid. MS-ESI: [M+H]+ 340.1 Example 231b (4-Chloro(10-fluorooxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl Acetate 231b A round-bottomed flask was charged with 231a (600 mg, 1.72 mmol), 20 dichloromethane (50 mL), and triethylamine (5 mL). The solution was stirred at 0oC for 0.5 h and acetyl chloride (135 mg, 1.72 mmol) was added slowly. The mixture was stirred at 0oC for another 2.5 h. It was then evaporated under reduced pressure. The residue was ed by -gel column tography eluting with 9:1 petroleum ether/ethyl acetate to afford 231b (605 mg, 90%) as a yellow solid. MS-ESI: [M+H]+ 392.1 25 Example 231c 3-(Acetoxymethyl)(10-fluorooxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a] indol-2(1H)-yl)pyridinylboronic Acid 231c A 100-mL single-neck round-bottomed flask ed with a magnetic stirrer and a reflux condenser was charged with 231b (600 mg, 1.53 mmol), Pin2B2 (1.94 g, 7.65 mmol), PdCl2(dppf) (65 mg, 0.080 mmol), X-Phos (73 mg, 0.15 mmol), ium acetate (304 mg, 30 3.1 mmol), and dioxane (30 mL). After three cycles of /argon flush, the mixture was heated at 65ºC for 5 h. It was then cooled to room temperature and filtered. The filtrate was 328 trated under reduced pressure and the resulting residue was washed with eum ether to afford 231c (412 mg, 67%) as yellow solid. MS-ESI: [M+H]+ 402.1 Example 231d (S)-(2-(10-Fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)-4 -(1-methyl(5-(2-methyl(oxetanyl)piperazinyl)pyridin 5 ylamino)oxo-1,6-dihydropyridazinyl)pyridinyl)methyl Acetate 231d A round-bottomed flask equipped with a reflux condenser was charged with 231c (200 mg, 0.50 mmol), (S)chloromethyl(5-(2-methyl(oxetanyl)piperazin yl)pyridinylamino)pyridazin-3(2H)-one 224c (195 mg, 0.50 mmol), PdCl2(dppf) (24 mg, 10 0.030 mmol), K3PO4 (212 mg, 1.0 mmol), sodium acetate (98 mg, 1.0 mmol), acetonitrile (30 mL), and water (3 mL). After three cycles of /argon flush, the mixture was heated at 100 oC for 2 h. It was then cooled to room temperature and filtered. The filtrate was ated under reduced pressure. The residue was purified with silica-gel column chromatography eluting with 1:3 petroleum/ethyl acetate to afford 231d as a yellow solid 15 (213 mg, 60%). MS-ESI: [M+H]+ 712.3 Example 231 10-fluoro[3-(hydroxymethyl)[1-methyl[[5-[(2S)methyl (oxetanyl)piperazinyl]pyridyl]amino]oxo-pyridazinyl]pyridyl]-3,4,6,7,8,9- dropyrazino[1,2-a]indolone 231 A mixture of 231d (150 mg, 0.21 mmol) and lithium hydroxide (51 mg, 2.1 mmol) in 20 i-propanol/THF (1:1, 10 mL) and water (3 mL) was stirred at room temperature for 1 h. The mixture was evaporated under reduced pressure and the residue was extracted with ethyl acetate (2 X 10 mL). The combined ethyl acetate extract was trated under reduced pressure and the residue was purified with reverse-phase prep-HPLC to afford 231 (83 mg, 59%) as a yellow solid. MS-ESI: [M+H]+ 670.3. 1H NMR (500 MHz, 6) δ 9.29 (s, 25 1H), 8.53 (d, J = 5.5 Hz, 1H), 8.42 (s, 1H), 7.91 (s, 1H), 7.43-7.40 (m, 3H), 4.76 (bs, 1H), 4.60-4.54 (m, 3H), 4.49-4.46 (m, 1H), 4.43-4.37 (m, 2H), 4.21-4.56 (m, 2H), 4.07-4.03 (m, 1H), 3.89-3.83 (m, 2H), 3.78 (s, 3H), 3.41-3.37 (m, 1H), 3.22-3.19 (m, 1H), 3.00-2.93 (m, 329 1H), 2.65-2.60 (m, 2H), 2.55-2.54 (m, 1H), 2.43-2.39 (m, 3H), 2.27-2.24 (m, 1H), .07 (m, 1H), 1.76-1.66 (m, 4H), 0.97 (d, J = 9.0 Hz, 3H).
Example 232a o(1,2,3,6-tetrahydropyridinyl)pyridine 232a 5 A mixture of tert-butyl 4-(6-nitropyridinyl)-5,6-dihydropyridine-1(2H)-carboxylate 200a (2.0 g, 6.6 mmol) in HCl/dioxane (20 mL, 4M) was stirred at room temperature for 2 hours. It was then evaporated under reduced pressure. The residue was washed with ethyl acetate (3 X 7 mL) to afford 232a as a yellow solid (1.0 g, 74%). MS-ESI: [M+H]+ 206. 10 Example 232b 5-(1-Methyl-1,2,3,6-tetrahydropyridinyl)nitropyridine 232b To a solution of 232a (1.2 g, 5.8 mmol) in CH3OH (25 mL) was added HCHO (1 mL, 35 mmol) and acetic acid (1 mL), followed by the addition of NaBHCN3 (1.0 g, 12 mmol).
The mixture was stirred at room ature for 2 h. It was then evaporated under reduced pressure and the residue was purified by reverse-phase Combiflush eluting with 0.3% 15 NH4HCO3 in water/acetonitrile to afford 232b as a yellow solid (1.0 g, 78%). : [M+H]+ 220.
Example 232c 5-(1-Methylpiperidinyl)pyridinamine 232c A 250-mL single-neck round-bottomed flask was purged with nitrogen and d with 232b (2.0 g, 9.0 mmol), 10% palladium on carbon (50% wet, 200 mg), and methanol (40 20 mL). The flask was evacuated, charged with hydrogen gas, and stirred under hydrogen at room temperature for 12 h. The hydrogen was then evacuated and nitrogen was charged to the flask. The st was removed by filtration through a pad of CELITE® and the filtrate was concentrated under reduced pressure to afford 232c (1.6 g, 92.5%), which was used ly in the next step without further purification. MS-ESI: [M+H]+ 192 330 Example 232d 5-Bromomethyl(5-(1-methylpiperidinyl)pyridin ylamino)pyridine-2(1H)-one 232d A 100-mL single-neck round-bottomed flask ed with a magnetic stirrer and a reflux condenser was charged with 232c (1.5 g, 7.9 mmol), 3,5-dibromomethylpyridin- 5 2(1H)-one (2.0 g, 7.9 mmol), cesium carbonate (5.0 g, 16 mmol), and 1,4-dioxane (50 mL).
After bubbling nitrogen through the resulting suspension for 30 minutes, os (455 mg, 0.79 mmol) and tris(dibenzylideneacetone)dipalladium(0) (718 mg, 0.79 mmol) were added.
The system was subject to three cycles of vacuum/argon flush and heated at 100℃ for 2 h.
After this time the reaction was cooled to room temperature and filtered. The te was 10 partitioned between ethyl acetate (100 mL) and water (100 mL). The aqueous layer was separated and extracted with ethyl acetate (3 X 50 mL). The combined organic layer was washed with brine (3 X 20 mL) and dried over sodium sulfate. The drying agent was removed by filtration and the filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column tography g with 1:3 ethyl acetate/petroleum 15 ether to afford 232d as a brown solid (1.5 g, 50%). MS-ESI: [M+H]+ 377.
Example 232e (4-(1-Ethyl(5-(1-methylpiperidinyl)pyridinylamino) oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- yl)pyridinyl)methyl Acetate 232e A round-bottomed flask equipped with a reflux condenser was d with 232d 20 (160 mg, 0.40 mmol), toxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)pyridinylboronic acid 113i (191 mg, 0.50 mmol), Pd(dppf)Cl2 (20 mg, 0.024 mmol), K3PO4 (180 mg, 0.80 mmol), sodium acetate trihydrate (120 mg, 0.80 mmol), water (0.5 mL), and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100 oC for 2 h. It was then filtered and the filtrate was evaporated in 25 vacuo. The residue was purified by -gel column chromatography eluting with 25:1 dichloromethane/CH3OH to afford 232e as a yellow solid (180 mg, 55%). MS-ESI: [M+H]+ 636.3 Example 232 2-[3-(hydroxymethyl)[1-methyl[[5-(1-methylpiperidyl) pyridyl]amino]oxopyridyl]pyridyl]-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indolone 30 232 A mixture of 232e ( 180 mg, 0.30 mmol) and lithium hydroxide (130 mg, 3.0 mmol) in THF/ i-propanol (6:3, 9 mL) and water (3 mL) was stirred at 30 oC for 1 h. The mixture was evaporated under reduced pressure and the residue was extracted with ethyl acetate (2 X 20 mL). The combined ethyl acetate extract was concentrated under reduced pressure and the 331 residue was purified by reverse-phase prep-HPLC to 232 (55 mg, 35 %) as white solid. MSESI : [M+H]+ 594.3. 1H NMR (500 MHz, CDCl3) δ 8.72 (d, J = 3.0 Hz, 1H), 8.51 (d, J = 6.0 Hz, 1H), 8.11 (d, J = 3.0 Hz, 1H), 7.88 (d, J = 2.0 Hz, 1H), 7.44-7.36 (m, 2H), 6.90 (s, 1H), 6.80-6.78 (m, 1H), 5.08-5.04 (m, 1H), 4.64-4.50 (m, 2H), 4.34-4.29 (m, 1H), 4.16-4.10 (m, 5 2H), 3.91-3.87 (m, 1H), 3.72 (s, 3H), 3.03-3.00 (m, 2H), 2.62-2.56 (m, 4H), 2.46-2.42 (m, 1H), 2.36 (s, 1H) 2.13-2.07 (m, 2H), 1.92-1.82 (m, overlap, 8H).
Example 233a tert-Butyl 2-(5-Bromomethyloxo-1,2-dihydropyridin ylamino)-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate 233a 10 A 100-mL round-bottomed flask equipped with a reflux condenser was d with tert-butyl 2-amino-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (600 mg, 2.35 mmol), 3,5-dibromomethylpyridin-2(1H)-one (942 mg, 3.53 mmol), Pd2(dba)3 (214 mg, 0.235 mmol), Xantphos (270.5 mg, 0.47 mmol), Cs2CO3 (1.53 g, 4.7 mmol), and dioxane (30 mL). After ng nitrogen through the mixture for 30 s, it was heated at 110oC 15 under N2 tion for 12 h. Analysis of reaction e by LCMS showed complete conversion to the desired product. The mixture was cooled to room temperature and filtered.
The filtrate was concentrated under reduced pressure and the residue was washed with acetonitrile to afford 233a (600 mg, 54%) as yellow solid. MS-ESI: [M+H]+ 441.1 332 Example 233b utyl 2-[(5-{3-[(Acetoxy)methyl]{4,4-dimethyloxo- 1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}methyloxo-1,2- dihydropyridinyl)amino]-4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridinecarboxylate 233b A 50-mL round-bottomed flask equipped with a reflux condenser was d with 5 233a (300 mg, 0.68 mmol), (3-(acetoxymethyl)(7,7-dimethyloxo-3,4,7,8-tetrahydro- 1H-cyclopenta[4,5]pyrrolo-[1,2-a]pyrazin-2(6H)-yl)pyridinyl)boronic acid 199e (1.8 g, 2.72 mmol), Pd(dppf)Cl2 (27.7mg , 0.034 mmol), K3PO4 (288.3 mg, 1.36 mmol), sodium e (111.5 mg, 1.36 mmol), water (10 drops), and acetonitrile (10 mL). After bubbling nitrogen through the mixture for 30 minutes, it was heated at 100oC under N2 protection for 1 10 h. Analysis of reaction mixture by LCMS showed complete conversion to the desired t.
The mixture was filtered and the filtrate was concentrated under reduced pressure and the residue was purified by silica-gel column tography eluting with 100:1 romethane/methanol to afford 233b (220 mg, 45%) as a yellow solid. MS-ESI: [M+H]+ 714.3 15 Example 233c (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}[1-methyloxo({4H,5H,6H,7H-[1,3]thiazolo[5,4-c]pyridin no)-1,6-dihy-dropyridinyl]pyridinyl)methyl Acetate 233c To a solution of 233b (220 mg, 0.308 mmol) in ethyl acetate (5 mL) was added a solution of HCl in ethyl acetate (0.123 mL, 2.5M, 0.308 mmol). The mixture was stirred at 20 room temperature for 1 h. It was then concentrated under reduced pressure to afford 233c (180 mg, crude), which was used directly for next step without further purification. MS-ESI: [M+H]+ 614.3 Example 233 3-[3-(hydroxymethyl)[1-methyloxo(4,5,6,7- tetrahydrothiazolo[5,4-c]pyridinylamino)pyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- 25 tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 233 To a solution of 233c (180 mg, 0.29 mmol) in THF (3 mL) and ol (3 mL) was added water (1 mL) and lithium hydroxide (14.0 mg, 0.58 mmol). The reaction mixture was stirred at room temperature for 1 h. It was then concentrated under reduced pressure and the resulting residue was purified by reverse-phase prep-HPLC to afford 233 (28.6 mg, 17%) 30 as white solid. MS-ESI: [M+H]+ 572.3. 1H NMR (500 MHz, 6) δ 9.85 (s, 1H), 8.63 (d, J = 2.5 Hz, 1H), 8.48 (d, J = 6.5 Hz, 1H), 7.57 (d, J = 2.5 Hz, 1H), 7.32 (d, J = 6.0 Hz, 1H), 6.55 (s, 1H), 4.95-4.92 (m, 1H), 4.47-4.37 (m, 2H), 4.25-4.18 (m, 3H), 3.86-3.84 (m, 1H), 3.71 (s, 2H), 3.59 (s, 3H), 2.92-2.90 (m, 2H), 2.62-2.56 (m, 2H), 2.50-2.39 (m, 5H), 1.21 (s, 6H). 333 Example 235a 4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- dienyl}[1-methyl({5-[(2S)methyl(oxetanyl)piperazinyl]pyrazin yl}amino)oxo-1,6- dihydropyridinyl]pyridinyl)methyl Acetate 235a 5 A 50-mL -neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with (S)bromomethyl(5-(2-methyl(oxetanyl) piperazinyl)pyrazinylamino)pyridin-2(1H)-one 230f (200 mg, 0.46 mmol), {3- [(acetoxy)methyl]{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]-dodeca-2(6),7-dien- 10-yl}pyridinyl}boronic acid 199e (366 mg, 0.92 mmol), Pd(dppf)Cl2 (38 mg, 0.046 10 mmol), sodium acetate (126 mg, 0.92 mmol), K3PO4 (196 mg, 0.92 mmol), water (0.5 mL), and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the e was heated at 65oC for 3 hrs. After cooling to room temperature the reaction was filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with romethane/methanol (50:1 to 30:1) to afford 235a 15 as brown solid (100 mg, 31%). MS-ESI: [M+H]+ 708.5. e 235 3-[3-(hydroxymethyl)[1-methyl[[5-[(2S)methyl(oxetan yl)piperazinyl]pyrazinyl]amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 235 A mixture of 235a (90.0 mg, 0.13 mmol) and lithium hydroxide (36.4 mg, 3.25 mmol) 20 in i-propanol /THF (1:1, 5 mL) and water (1 mL) was stirred at 35oC for 0.5 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by reverse-phase prep-HPLC to afford 235 (18.2 mg, 22%). MS-ESI: [M+H]+ 666.3. 1H NMR (500 MHz, DMSO-d6) δ 8.61 (s, 1H), 8.48 (d, J = 4.5 Hz, 1H), 8.39 (d, J = 1.5 Hz, 1H), 8.34 (s, 1H), 7.83 (s, 1H), 7.47 (d, J = 2.0 Hz, 1H), 25 7.33 (d, J = 5.0 Hz, 1H), 6.56 (s, 1H), 4.96 (bs, 1H), .55 (m, 2H), 4.49-4.47 (m, 1H), 4.42-4.37 (m, overlap, 4H), 4.22-4.18 (m, overlap, 3H), 3.84-3.76 (m, 2H), 3.60 (s, 3H), 3.39- 3.37 (m, 1H), 3.02-2.81 (m, 1H), 2.78-2.76 (m, 1H), 2.62-2.56 (m, 3H), 2.43-2.41 (m, 2H), 2.10-2.07 (m, 1H), 1.92-1.90 (m, 1H), 1.22 (s, 6H), 1.12 (d, J = 6.0 Hz, 3H). 334 Example 236a (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- dienyl}(1-methyl{[5-(1-methylpiperidinyl)pyridinyl]amino}oxopyridin yl)pyridinyl)methyl Acetate 236a N N NH OAc O N N N O N 236a 5 A 50-mL round-bottomed flask equipped with a reflux condenser was charged with 5- bromomethyl(5-(1-methylpiperidinyl)pyridinylamino)pyridin-2(1H)-one 232d (160 mg, 0.40 mmol), {3-[(acetoxy)methyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (240 mg, 0.60 mmol), Pd(dppf)Cl2 (20 mg, 0.020 mmol), K3PO4 (180 mg, 0.80 mmol), sodium acetate 10 rate (120 mg, 0.80 mmol), water (0.5 mL), and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 2 h. It was then filtered and the filtrate was evaporated under reduced pressure. The residue was purified on silica-gel column chromatography eluting with 25:1 dichloromethane/methanol to afford 236a as a yellow solid (150 mg, 38%). MS-ESI: [M+H]+ 650.3 15 Example 236 3-[3-(hydroxymethyl)[1-methyl[[5-(1-methylpiperidyl) pyridyl]amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- ydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 236 A mixture of 236a (150 mg, 0.25 mmol) and lithium ide (105 mg, 2.5 mmol) in THF/i-propanol (6:3, 9 mL) and water (3 mL) was d at 30oC for 1 h. The mixture was 20 evaporated in vacuo and the residue was extracted with ethyl acetate (2 X 20 mL). The combined ethyl acetate extract was concentrated under reduced pressure and the e was purified by reverse-phase prep-HPLC to afford 236 (40 mg, 30%) as light green solid. MSESI : [M+H]+ 608.3. 1H NMR (500 MHz, CDCl3) δ 8.73 (s, 1H), 8.52 (d, J = 4.0 Hz, 1H), 8.13 (s, 1H), 7.90-7.88 (m, 2H), 7.45-7.37 (m, 2H), 6.86-6.80 (m, 2H), 5.11 (bs, 1H), 4.67- 25 4.65 (m, 1H), 4.53-4.51 (m, 1H), 4.35-4.33 (m, 1H), 4.18 (bs, 2H), 3.90-3.89 (m, 1H), 3.73 (s, 3H), 3.24-3.22 (m, 2H), 2.59-2.50 (m, , p, 8H), 2.36-2.32 (m, 2H), 2.01-1.87 (m, 4H), 1.29 (s, 6H). 335 Example 237a 2-{10-Fluorooxo-1H,2H,3H,4H,6H,7H,8H,9H-pyrido[3,4- b]indolizinyl}[1-methyl({5-[(2S)methyl(oxetanyl)piperazinyl]pyridin yl}amino)oxo-1,6-dihydropyridinyl]pyridinecarbaldehyde 237a O N N N NH N O O N N F O N 237a 5 A 50-mL single-neck round-bottomed flask ed with a magnetic r and a reflux condenser was charged with 4-chloro(10-fluorooxo-3,4,6,7,8,9-hexahydropyrido ]indolizin-2(1H)-yl)nicotinaldehyde 134c (70 mg, 0.20 mmol), 1-methyl- 3-({5-[(2S)- 2-methyl(oxetanyl)piperazinyl]pyridinyl}amino)(tetramethyl-1,3,2- orolanyl)-1,2-dihydropyridinone 191j (192 mg, 0.40 mmol), Pd(dppf)Cl2 (33 mg, 10 0.040 mmol), potassium acetate (39 mg, 0.40 mmol), K3PO4 (87 mg, 0.40 mmol), acetonitrile (7 mL), and water (6 drops). After three cycles of vacuum/argon flush, the mixture was heated at 95oC for 2 h. It was then filtered and the filtrate was evaporated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 50:1 ethyl acetate/methanol to afford 237a (286 mg, purity: 46%, yield: 98%) as a solid. MS-ESI: 15 [M+H]+ 667.3 Example 237 10-fluoro[3-(hydroxymethyl)[1-methyl[[5-[(2S)methyl (oxetanyl)piperazinyl]pyridyl]amino]oxopyridyl]pyridyl]-3,4,6,7,8,9- hexahydropyrido[3,4-b]indolizinone 237 To a solution of 237a (131 mg, 0.197 mmol) in methanol (7 mL) was added sodium 20 borohydride (59.8 mg, 1.57 mmol) at 0oC. The reaction was stirred at 0-25oC for 1.5 h. It was then quenched with water (1.5 mL). The mixture was evaporated under reduced pressure and the residue was extracted with dichloromethane (3 X 30 mL). The combined dichloromethane t was concentrated under reduced pressure and the residue was purified by reversephase prep-HPLC to afford 237 (36 mg, 28%) as a white solid. : [M+H]+ 669.3. 1H 25 NMR (500 MHz, CDCl3) δ 8.66 (d, J = 1.5 Hz, 1H), 8.51 (d, J = 5.0 Hz, 1H), 7.99 (s, 1H), 7.85 (s, 1H), 7.83 (d, J = 2.0 Hz, 1H), 7.36-7.32 (m, 2H), 6.84 (d, J = 9.0 Hz, 1H), 4.90-4.86 (m, 1H), 4.73-4.65 (m, 5H), 4.39-4.30 (m, 2H), .77 (m, 3H), 3.72 (s, 3H), 3.56-3.48 (m, 336 2H), .09 (m, 2H), 2.98-2.92 (m, 2H), 2.79-2.75 (m, 2H), 2.59-2.57 (m, 1H), 2.50-2.49 (m, 2H), 2.22-2.21 (s, 1H), 2.04-1.99 (m, 2H), 1.88-1.84 (m, 2H), 1.01 (d, J = 6.0 Hz, 3H).
Example 238a (E)-Methyl 3-((tert-Butylsulfinylimino)methyl)-5,6,7,8- tetrahydroindolizinecarboxylate 238a O O NaBH4, S O S H2N MeOH S O N N N rt, 1 h N N OMe H OMe KHSO4, DCM, rt, OMe 10 h O O O 238a 238b 1) 4 eq. LHMDS N NH2 THF, -78°C, 10 min, N HCl/ether 2) rt for 2 h rt. 4 h OMe NH O O 238c 238d O N O N Br Cl N N NH N O N O 191j O N Cl N N Pd2(dba)3, xantphos, Pd(dppf)Cl2, Cs2CO3, dioxane, O N NaOAc, O N 90°C, 16 h K3PO4, CH3CN, H2O, 85°C, 2 h 5 238e 238f A 500-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with methyl 3-formyl-5,6,7,8-tetrahydroindolizinecarboxylate (10.0 g, 48.3 mmol, 1.0 eq.), 2-methylpropanesulfinamide (11.7 g, 96.6 mmol, 2.0 eq.), KHSO4 (32.8 g, 241.5 mmol, 5 eq.), and romethane (250 mL). The mixture was stirred at room ature 10 for 10 h. It was then filtered and filtrate was concentrated under reduced pressure. The resulting residue was purified by -gel column chromatography eluting with 1:3 ethyl acetate/petroleum ether to afford 238a (12.4 g, 83%) as a yellow solid. MS: [M+H]+ 311.3.
Example 238b Methyl 3-((1,1-Dimethylethylsulfinamido)methyl)-5,6,7,8- tetrahydroindolizinecarboxylate 238b 15 A 250-mL single-neck round-bottomed flask equipped with a ic stirrer was charged with 238a (4.0 g, 12.8 mmol, 1.0 eq.), NaBH4 (2.9 g, 76.9 mmol, 6.0 eq.), and 337 methanol (100 mL). The on mixture was stirred at room temperature for 1 h. After this time water (50 mL) was added to the reaction and the resulting mixture was concentrated under reduced re. The residue was extracted with romethane (3 X 50 mL). The combined organic layer was evaporated under reduced pressure to afford 238b (3.9 g, 96%), 5 which was directly used in next step without further purification. MS: [M-C4H10NOS]+ 192.3.
Example 238c Methyl 3-(Aminomethyl)-5,6,7,8-tetrahydroindolizinecarboxylate 238c A 100-mL single-neck bottomed flask equipped with a magnetic stirrer was charged with 238b (3.5 g, 11.2 mmol), saturated HCl/diethyl ether solution (15 mL), and 10 dichloromethane (15 mL). The mixture was stirred at room temperature for 4 h. After the reaction was completed, ted aqueous NaHCO3 on (50 mL) was added to the on mixture and the mixture was extracted with dichloromethane (3 X 50 mL). The combined organic layer was evaporated under reduced pressure to afford 238c (2.2 g, 94%), which was directly used in the next step without further purification. MS: [M-NH2]+ 192.1. 15 1H NMR (500 MHz, MeOD) δ 6.28 (s, 1H), 4.38 (s, 2H), 4.03 (t, J = 6.5 Hz, 2H), 3.84 (s, 3H), 2.78 (t, J = 6.5 Hz, 2H), 2.06-2.02 (m, 2H), 1.87-1.82 (m, 2H).
Example 238d 2,3,5,6,7,8-Hexahydropyrrolo[3,4-b]indolizinone 238d A 100-mL -neck round-bottomed flask equipped with a ic stirrer was charged with 238c (1.3 g, 6.25 mmol, 1.0 eq.) and THF (20 mL). At -78 ºC, to the solution 20 was added lithium hexamethyldisilazane/THF (18.7 mL, 18.7 mmol, 3.0 eq.). It was then stirred at room temperature for 2 hrs. After the reaction was completed, saturated aqueous NH4Cl solution (30 mL) was added and the mixture was concentrated under d pressure.
The residue was extracted with dichloromethane (3 X 50 mL) and the combined organic layer was evaporated under reduced pressure. The resulting residue was ed by silica-gel 25 column chromatography eluting with 60:1 dichloromethane/methanol to afford 238d (585 mg, 53%) as a yellow solid. MS: [M+H]+ 177.3. 1H NMR (500 MHz, DMSO-d6) δ 7.38 (s, 1H), 5.82 (s, 1H), 4.13 (s, 2H), 3.86 (t, J = 6.5 Hz, 2H), 2.73 (t, J = 6.5 Hz, 2H), 191-1.88 (m, 2H), 1.75-1.73 (m, 2H).
Example 238e 4-Chloro(1-oxo-5,6,7,8-tetrahydro-1H-pyrrolo[3,4-b]indolizin- 30 2(3H)-yl)nicotinaldehyde 238e A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 238d (400 mg, 2.27 mmol), 2-bromo chloronicotinaldehyde (1.50 g, 6.90 mmol, 3.0 eq.), Pd2(dba)3 (208 mg, 0.227 mmol, 0.1 eq.), xantphos (131 mg, 0.227 mmol, 0.1 eq.), Cs2CO3 (1.50 g, 4.54 mmol, 2.0 eq.), and dioxane 338 (30 mL). After bubbling nitrogen through the resulting mixture for 30 minutes, the reaction mixture was stirred at 90 ºC for 16 h. After the reaction was complete, the mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the residual was purified by silica-gel column chromatography eluting with 1:3 ethyl 5 e/petroleum ether to afford 238e (300 mg, 42%) as a light yellow solid. : [M+H]+ 316.1. e 238f S)(1-methyl(5-(2-methyl(oxetanyl)piperazinyl)pyridin- 2-yl-amino)oxo-1,6-dihydropyridinyl)(1-oxo-5,6,7,8-tetrahydro-1H-pyrrolo[3,4- b]indolizin-2(3H)-yl)nicotinaldehyde 238f 10 A 50-mLsingle-neck bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with 238e (150 mg, 0.48 mmol, 1.0 eq.), (S)methyl(5-(2- methyl(oxetanyl)piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2- dioxaborolanyl)pyridin-2(1H)-one 191j (459 mg, 0.95 mmol, 2.0 eq.), Pd(dppf)Cl2 (39 mg, 0.048 mmol, 0.1 eq.), sodium acetate (78 mg, 0.95 mmol, 2.0 eq.), K3PO4 (202 mg, 0.95 15 mmol, 2.0 eq.), acetonitrile (10 mL), and water (1 mL). After three cycles of vacuum/argon flush, the mixture was heated at 85 ºC for 2 h. It was then cooled to room temperature and filtered. The te was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 40:1 dichloromethane/ethanol to afford 238f (90 mg, 30%) as a yellow solid. MS-ESI: [M+H]+ 635.3. 20 Example 238 hydroxymethyl)[1-methyl[[5-[(2S)methyl(oxetan yl)piperazinyl]pyridyl]amino]oxopyridyl]pyridyl]-5,6,7,8-tetrahydro-1H- pyrrolo[3,4-b]indolizinone 238 A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with 238f (90 mg, 1.0 eq., 0.14 mmol), NaBH4 (23 mg, 5 eq., 0.60 mmol), and 25 methanol (5 mL). The resulting mixture was stirred at room temperature for 20 s and quenched with water. It was then concentrated under reduced re and the residue was purified by reverse-phase prep-HPLC to afford 238 (60 mg, 66%) as a white solid. MS-ESI: [M+H]+ 637.3. 1H NMR (500 MHz, CDCl3) δ 8.69 (d, J = 1.5 Hz, 1H), 8.47 (d, J = 5.0 Hz, 1H), 7.99 (d, J = 2.5 Hz, 1H), 7.93 (d, J = 2.0 Hz, 1H), 7.86 (s, 1H), 7.36-7.31 (m, 2H), 6.83 30 (d, J = 9.0 Hz, 1H), 6.15 (s, 1H), 5.71 (t, J = 6.5 Hz, 1H), 4.96 (s, 2H), 4.72-4.63 (m, 4H), 4.52-4.51 (m, 2H), 3.97-3.95 (m, 2H), 3.75 (s, 3H), 3.55-3.53 (m, 1H), 3.48-3.46 (m, 1H), 3.10-3.08 (m, 2H), 2.91-2.89 (m, 2H), 2.58-2.56 (m, 1H), 2.49-2.48 (m, 2H), 2.23-2.19 (m, 1H), 2.08-2.02 (m, 2H), 1.93-1.89 (m, 2H), 1.00 (d, J = 6.0 Hz, 3H). 339 Example 239a tert-Butyl 5-Bromomethyloxo-1,2-dihydropyridin ylamino)pyridineyl)azetidinecarboxylate 239a Br HCl O Boc N HN Boc N N Br N NH oxane, rt, 4 h N NH Pd2(dba)3, Xantphos, O O N NH2 Cs2CO3, dioxane N N 105 °C, 0.5 h Br Br 239a 239b N N NH 50 equiv. HCHOaq, AcO O AcOH, )3BH N MeOH, rt, 2 h N NH 113i N N O PdCl2(dppf), AcONa,K3PO4, O N N Br CH3CN, H2O, 100 oC, 2 h 239d 239c A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 5 reflux condenser was charged with 1,4-dioxane (50 mL), tert-butyl 3-(6-aminopyridin yl)azetidinecarboxylate (1.8 g, 7.2 mmol), 3,5-dibromomethylpyridin-2(1H)-one (1.9 g, 7.2 mmol), and cesium carbonate (4.7 g, 14.4 mmol). After bubbling nitrogen through the resulting suspension for 30 minutes, XantPhos (418 mg, 0.72 mmol) and tris(dibenzylideneacetone)dipalladium(0) (661 mg, 0.72 mmol) were added. The on 10 mixture was subjected to three cycles of vacuum/argon flush and heated at 105ºC for 0.5 h.
After this time the reaction was cooled to room temperature and filtered. The filtrate was ioned between ethyl acetate (120 mL) and water (60 mL). The aqueous layer was separated and extracted with ethyl acetate (3 X 80 mL). The combined organic layer was washed with brine (30 mL) and dried over sodium sulfate. The drying agent was removed by 15 filtration and the filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column tography eluting with 1:4 ethyl acetate/petroleum ether to afford the 239a as a yellow solid (3.06 g, 98%). MS-ESI: [M+H]+ 435.
Example 239b 3-(5-(Azetidinyl)pyridinylamino)bromomethylpyridin- 2(1H)-one 239b 20 Compound 239a (1.0 g, 2.3 mmol) was suspended in 4M HCl/dioxane (10 mL). The reaction mixture was d at room temperature for 4 h. It was then concentrated under 340 reduced re. The residue was basified with aqueous NaOH and the resulting mixture was extracted with dichloromethane. The combined c layer was washed with brine and concentrated under reduced pressure to afford 239b as a yellow solid (650 mg, 84%). MSESI : [M+H]+ 335. 5 Example 239c 5-Bromomethyl(5-(1-methylazetidinyl)pyridin ylamino)pyridine-2(1H)-one 239c A mixture of 239b (469 mg, 1.4 mmol), 37% s formaldehyde (4.0 g, 50 mmol), NaBH3CN (261 mg, 4.2 mmol), and 1M zinc chloride in ethoxyethane (4 mL, 4.2 mmol) in methanol (40 mL) was stirred at room temperature for 2 hours. The e was added to 10 water (20 mL) and concentrated under reduced pressure. The residue was extracted with dichloromethane (3 X 50 mL). The combined organic layer was concentrated under reduced pressure and the residue was purified by silica-gel column chromatography eluting with 10:1 methylene de/methanol to afford 239c as a yellow solid (300 mg, 83%). MS-ESI: [M+H]+ 349. 15 Example 239d (4-(1-Methyl(4-(1-methylazetidinyl)phenylamino)oxo-1,6- dihydro-pyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridin- 3-yl)methyl Acetate 239d A 50-mL round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 239c (106 mg, 0.30 mmol), 3-(acetoxymethyl)(1-oxo- 20 3,4,6,7,8,9-hexa-hydropyrazino[1,2-a]indol-2(1H)-yl)pyridinylboronic acid 113i (115 mg, 0.30 mmol), Pd(dppf)Cl2 (25 mg, 0.030 mmol), K3PO4 (127 mg, 0.60 mmol), sodium acetate (49 mg, 0.60 mmol), water (1 mL) and itrile (10 mL). After three cycles of vacuum/argon flush, the mixture was heated under reflux for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the 25 resulting e was purified by -gel column chromatography eluting with 15:1 dichloromethane/methanol to afford 239d as white solid (100 mg, 49%). MS-ESI: [M+H]+ 607.3 Example 239 2-[3-(hydroxymethyl)[1-methyl[[5-(1-methylazetidinyl) l]amino]oxopyridyl]pyridyl]-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indolone 30 239 To a solution of 239d (100 mg, 0.16 mmol) in propanol (2 mL), tetrahydrofuran (2 mL), and water (1 mL) was added lithium hydroxide (38 mg, 1.60 mmol). The mixture was stirred at 30oC for 2 h. It was then evaporated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 239 (22.5 mg, 26%) as a white solid. MS-ESI: 341 [M+H]+ 566.4. 1H NMR (500 MHz, CDCl3) δ 8.75 (d, J = 1.5 Hz, 1H), 8.52 (d, J = 5.0 Hz, 1H), 8.17 (d, J = 2.0 Hz, 1H), 7.91-7.89 (m, 2H), 7.59 (d, J = 8.5 Hz, 1H), 7.38 (d, J = 5.0 Hz, 1H), 6.91 (s, 1H), 6.84 (d, J = 8.5 Hz, 1H), 5.09-5.06 (m, 1H), 4.66-4.64 (m, 1H), 4.54- 4.50 (m, 1H), 4.36-4.33 (m, 1H), 4.18-4.11 (m, 2H), 3.92-3.88 (m, 1H), 3.75-3.72 (m, overlap, 5 5H), 3.63-3.58 (m, 1H), 3.16-3.14 (m, 2H), 2.64-2.58 (m, 4H), 2.40 (s, 3H), .90 (m, 2H), 1.84-1.79 (m, 2H).
Example 240a (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- dienyl}(1-methyl{[5-(1-methylazetidinyl)pyridinyl]amino}oxopyridin yl)pyridinyl)methyl Acetate 240 10 A 50-mL bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 5-bromomethyl(5-(1-methylazetidinyl)pyridin ylamino)-pyridin-2(1H)-one 239c (106 mg, 0.30 mmol), {3-[(acetyloxy)methyl]{4,4- dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic 15 acid 199e (119 mg, 0.30 mmol), Pd(dppf)Cl2 (25 mg, 0.031 mmol), K3PO4 (127 mg, 0.60 mmol), sodium acetate (49 mg, 0.60 mmol), water (0.5 mL), and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at reflux for 2 h. It was then cooled to room temperature and filtered. The te was concentrated under reduced pressure and the resulting e was purified by silica-gel column chromatography eluting with 15:1 20 romethane/methanol to afford 240a as white solid (80 mg, 48%). MS-ESI: [M+H]+ 622.7 Example 240 3-[3-(hydroxymethyl)[1-methyl[[5-(1-methylazetidinyl) pyridyl]amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 240 25 To a solution of 240a (80 mg, 0.130 mmol) in propanol (2 mL), tetrahydrofuran (2 mL), and water (1 mL) was added m hydroxide (38 mg, 1.60 mmol). The mixture was stirred at 30oC for 1 h. The reaction was evaporated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 240 24.3 mg, 33%) as a white solid. MS- 342 ESI: [M+H]+ 580.4. 1H NMR (500 MHz, CDCl3) δ 8.76 (d, J = 2.0 Hz, 1H), 8.52 (d, J = 5.0 Hz, 1H), 8.17 (d, J = 2.0 Hz, 1H), 7.92-7.90 (m, 2H), 7.59 (dd, J = 1.5, 8.5 Hz, 1H), 7.38 (d, J = 5.0 Hz, 1H), 6.87 (s, 1H), 6.84 (d, J = 8.5 Hz, 1H), 5.09-5.06 (m, 1H), 4.68-4.66 (m, 1H), 4.56-4.54 (m, 1H), 4.37-4.35 (m, 1H), 4.19-4.17 (m, 2H), 3.90-3.88 (m, 1H), 3.75-3.72 (m, 5 overlap, 5H), .62 (m, 1H), .16 (m, 2H), 2.60 (d, J = 5.0 Hz, 2H), 2.54 (s, 2H), 2.42 (s, 3H), 1.30 (s, 6H).
Example 241a 2-{10-Fluorooxo-1H,2H,3H,4H,6H,7H,8H,9H-pyrido[3,4- b]indolizinyl} hyl({5-methyl-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin yl}amino)oxo-1,6-dihydropyridinyl]pyridinecarbaldehyde 241a 10 A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 4-chloro(10-fluorooxo-3,4,6,7,8,9-hexahydropyrido [3,4-b]indolizin-2(1H)-yl)nicotinaldehyde 134c (59.6 mg, 0.17 mmol), 1-methyl ({5- -4H,5H,6H,7H-pyrazolo[1,5-a]pyrazinyl}amino)(tetramethyl-1,3,2-dioxa- 15 borolanyl)-1,2-dihydropyridinone 135a (261.8 mg, 0.68 mmol), Pd(dppf)Cl2 (25.0 mg, 0.030 mmol), Na2CO3 (54.1 mg, 0.51 mmol), DMF (6 mL), and water (0.75 mL). After three cycles of vacuum/argon flush, the e was heated at 70oC for 1 h. It was then filtered and the filtrate was evaporated under reduced pressure. The residue was purified by silica-gel column tography eluting with 50:1 dichloromethane/methanol to afford 241a (100 mg, 20 purity: 54%, yield: 56%) as a yellow solid. MS-ESI: [M+H]+ 571.3 Example 241 10-fluoro[3-(hydroxymethyl)[1-methyl[(5-methyl-6,7- dihydro-4H-pyrazolo[1,5-a]pyrazinyl)amino]oxopyridyl]pyridyl]-3,4,6,7,8,9- hexahydropyrido[3,4-b]indolizinone 241 To the solution of 241a (54.0 mg, 0.095 mmol) in ol (5 mL) was added sodium 25 borohydride (28.9 mg, 0.76 mmol) at 0oC. The reaction was stirred at 0-25oC for 1.5 h. It was then quenched with water (5 mL). The mixture was evaporated under reduced pressure and the residue was extracted with dichloromethane (3 X 30 mL). The combined dichloromethane extract was concentrated under reduced pressure and the residue was purified by reverse- 343 phase prep-HPLC to afford 241 (8.0 mg, 15%) as a white solid. MS-ESI: [M+H]+ 573.3. 1H NMR (500 MHz, DMSO-d6) δ 8.47 (d, J = 5.0 Hz, 1H), 8.21 (s, 1H), 8.06 (d, J = 2.0 Hz, 1H), 7.40(d, J = 2.0 Hz, 1H), 7.31 (d, J = 5.0 Hz, 1H), 5.88 (s, 1H), 4.87-4.85 (m, 1H), .35 (m, 2H), 4.13-4.08 (m, 1H), 3.92-3.89 (m, 3H), 3.79-3.76 (m, 2H), 3.58 (s, 3H), 3.49 (s, 2H), 5 2.99-2.94 (m, 2H), 2.79-2.77 (m, 2H), 2.66-2.64 (m, 2H), 2.35 (s, 3H), .78 (m, 2H), 1.75-1.73 (m, 2H).
Example 242a 3-(1,5-Dimethyl-1H-pyrazolylamino)methyl(4,4,5,5- tetramethyl-1,3,2 -dioxaborolanyl)pyridin-2(1H)-one 242a 10 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 5-bromo(1,5-dimethyl-1H-pyrazolylamino) pyridin-2(1H)-one 218a (800 mg, 2.69 mmol, 1.0 eq.), 4,4,4',4',5,5,5',5'-octamethyl- 2,2'-bi(1,3,2-dioxaborolane) (1.70 g, 6.73 mmol, 2.5 eq.), Pd2(dba)3 (123 mg, 0.13 mmol, 0.05 eq.), X-Phos (128 mg, 0.27 mmol, 0.1 eq.), potassium acetate (528 mg, 5.38 mmol, 2.0 15 eq.), and dioxane (50 mL). After three cycles of vacuum/argon flush, the mixture was heated at 70ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced re and the resulting residue was purified by silica-gel column chromatography eluting with 1:70 methanol/dichloromethane to afford 242a (740 mg, 79%) as a green solid. MS-ESI: [M+H]+ 345.3 20 e 242b 4-(5-(1,5-Dimethyl-1H-pyrazolylamino)methyloxo-1,6- dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin-2(1H)- yl)nicotinaldehyde 242b A-100 mL single-neck round-bottomed flask equipped with a ic stirrer and a reflux condenser was charged with 242a (282 mg, 0.82 mmol, 1.5 eq.), 4-chloro(1-oxo- 25 3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin-2(1H)-yl) nicotinaldehyde 139a (180 mg, 0.55 mmol, 1.0 eq.), Pd(dppf)Cl2 (45 mg, 0.055 mmol, 0.1 eq.), sodium acetate (90 mg, 1.25 mmol, 2.0 eq.), K3PO4 (232 mg, 1.25 mmol, 2.0 eq.), acetonitrile (20 mL), and water (1 mL). After three cycles of vacuum/argon flush, the mixture was heated at 90 ºC for 1 h. It was then 344 cooled to room temperature and filtered. The te was concentrated under d pressure and the resulting residue was purified by silica-gel column chromatography eluting with 50:1 romethane/methanol to afford 242b (150 mg, 48%) as a yellow solid. MS-ESI: [M+H]+ 512.3. 5 Example 242 5-[(1,5-dimethylpyrazolyl)amino]methyloxo l](hydroxymethyl)pyridyl]-3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizinone 242 A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with 242b (150 mg, 0.29 mmol, 1.0 eq.), NaBH4 (55 mg, 1.46 mmol, 5.0 eq.), and 10 methanol (10 mL). The resulting mixture was stirred at room temperature for 20 min. It was then filtered and the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase prep-HPLC to afford 242 (100 mg, 66%) as a white solid. MS-ESI: [M+H]+ 514.3. 1H NMR (500 MHz, DMSO-d6) δ 8.47 (d, J = 5.5 Hz, 1H), 8.04-8.03 (m, 2H), 7.42 (d, J = 2.0 Hz, 1H), 7.30 (d, J = 5.0 Hz, 1H), 6.05 (s, 1H), 5.90 (s, 1H), 4.87-4.85 (m, 15 1H), 4.43-4.35 (m, 2H), 4.18-4.12 (m, 1H), 3.98-3.93 (m, 1H), 3.84-3.78 (m, 2H), 3.59 (s, 3H), 3.58 (s, 3H), 3.06-2.92 (m, 2H), 2.75-2.68 (m, 2H), 2.18 (s, 3H), 1.94-1.92 (m, 2H), 1.79-1.73 (m, 2H).
Example 243a tert-Butyl 4-(Methylsulfonyloxy)piperidinecarboxylate 243a 345 To a solution of tert-butyl 4-hydroxypiperidinecarboxylate (14.0 g, 70.0 mmol) at 0oC in triethylamine (9.9 g, 98 mmol) and dichloromethane (100 mL) was added dropwise methanesulfonyl chloride (11.2 g, 98.0 mmol). The reaction was brought to ambient 5 temperature and stirred for 1 h. Then the on e was quenched with water (50 mL).
The aqueous layer was separated and extracted with ethyl acetate (2 X 50 mL). The combined organic layer was washed with brine (50 mL), and dried over sodium sulfate. The drying agent was removed by filtration and the filtrate was concentrated under reduced re to afford 243a, which was used in the next step without further purification (19.5 g, 100%). MS- 10 ESI: [M-t-Bu]+ 224.1 Example 243b tert-Butyl 4-(4-Nitro-1H-imidazolyl)piperidinecarboxylate 243b 346 A 250-mL single-neck round-bottomed flask ed with a magnetic stirrer and a reflux condenser was charged with 243a (7.0 g, 25.1 mmol), DMF (120 mL), 4-nitro-1H- imidazole (2.80 g, 25.1 mmol), and K2CO3 (6.9 g, 50.2 mmol). The mixture was heated at 120oC for overnight. After this time the reaction was cooled to room temperature and filtered. 5 The filtrate was evaporated in vacuo. The residue was purified by -gel column chromatography eluting with 2:2:1 ethyl acetate/petroleum ether/dichloromethane to afford 243b (2.4 g, 32.4%) as a yellow solid. MS-ESI: [M+H]+ 297.3 Example 243c tert-Butyl 4-(4-Amino-1H-imidazolyl)piperidinecarboxylate 243c 10 A 100-mL single-neck round-bottomed flask was purged with nitrogen and charged with 243b (2.3 g, 7.8 mmol), 10% palladium on carbon (10% wet, 230 mg), and l (40 mL). The mixture was evacuated, charged with hydrogen gas, and stirred at room temperature for 3 h. The hydrogen was then evacuated and nitrogen was d into the flask. The catalyst was removed by filtration through a pad of CELITE® and the filtrate was 15 concentrated under reduced pressure to afford 243c (2.0 g, 95 %). MS-ESI: [M+H]+ 267.2.
Example 243d tert-Butyl 4-(4-(5-Bromomethyloxo-1,2-dihydropyridin ylamino)-1H- imidazolyl)piperidinecarboxylate 243d A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with 243c (2.3 g, 8.6 mmol), 3,5-dibromomethylpyridin- 20 2(1H)-one (2.3 g, 8.6 mmol), tris-(dibenzylideneacetone)dipalladium(0) (789 mg, 0.86 mmol), XantPhos (994 mg, 1.72 mmol), Cs2CO3 (5.6 g, 17.2 mmol), and 1,4-dioxane (80 mL). After three cycles of vacuum/argon flush, the mixture was heated at 95oC for 4 hrs. It was then cooled to room ature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 25 dichloromethane/methanol (50:1 to 20:1) to afford 243d as yellow oil solid (2.3 g, 59%). MSESI : [M+H]+ 452.3. e 243e 5-Bromomethyl(1-(piperidinyl)-1H-imidazol ylamino)pyridin-2(1H)-one 243e A e of 243d (2.2 g, 4.88 mmol) and trifluoroacetic acid (20 mL) was stirred at 30 room temperature for 1 h. It was then concentrated under d pressure to afford crude 243e (1.5 g, 88 %), which was used in the next step without further purification. MS-ESI: [M+H]+ 352.2 e 243f 5-Bromomethyl(1-(1-(oxetanyl)piperidinyl)-1H-imidazol- 4-ylamino)pyridin-2(1H)-one 243f 347 A 250-mL single-neck round-bottomed flask equipped with a magnetic r and a reflux condenser was charged with 243e (2.2 g, 6.3 mmol), NaBH3CN (995 mg, 15.8 mmol), oxetanone (907 mg, 12.6 mmol), zinc chloride (2.1 g, 15.8 mmol), and ol (60 mL).
The reaction mixture was stirred at 50oC for 5 hrs and concentrated under reduced pressure. 5 To the e was added water and the resulting e was extracted with dichloromethane three times. The combined organic layer was then concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 1% triethylamine in methanol. The fractions containing the desired product were concentrated under reduced pressure. romethane was added to the residue and the 10 resulting suspension was filtered. The te was concentrated under reduced pressure afford 243f as a yellow solid (800 mg, 62%). MS-ESI: [M+H]+ 408.2 Example 243g (4-(1-Methyl(1-(1-(oxetanyl)piperidinyl)-1H-imidazol ylamino)oxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2- a]indol-2(1H)-yl)pyridinyl)methyl Acetate 243g 15 A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 243f (300 mg, 0.74 mmol), 3-(acetoxymethyl)(1-oxo- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinylboronic acid 113i (567 mg, 1.48 mmol), Pd(dppf)Cl2 (60.5 mg, 0.074mmol), K3PO4 (314 mg, 1.48 mmol), sodium acetate (201 mg, 1.48 mmol), water (0.5 mL), and acetonitrile (10 mL). After three cycles of 20 vacuum/argon flush, the mixture was heated at 90oC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under d pressure and the resulting residue was purified by silica-gel column chromatography eluting with 1:1 dichloromethane/methanol containing 0.5% triethylamine to afford 243g as yellow solid (100 mg, 20 %). MS-ESI: [M+H]+ 667.4. 25 Example 243 2-[3-(hydroxymethyl)[1-methyl[[1-[1-(oxetanyl) piperidyl]imidazolyl]amino]oxopyridyl]pyridyl]-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indolone 243 A mixture of 243g (80 mg, 0.12 mmol) and lithium hydroxide (100 mg, 2.4 mmol) in i-propanol/THF/water (2:2:1, 8 mL) was stirred at 35 oC for 30 mins. The reaction mixture 30 was then concentrated under d pressure. To the residue was added water and the resulting mixture was extracted with dichloromethane three times. The ed c layer was concentrated under reduced re and the resulting residue was purified by reverse-phase prep-HPLC to afford 243 (28.0 mg, 30%). MS-ESI: [M+H]+ 625.4. 1H NMR (500 MHz, DMSO-d6) δ 8.48 (d, J = 6.5 Hz, 1H), 7.54 (d, J = 1.5 Hz, 1H), 7.53 (s, 1H), 7.45 348 (d, J =3.0 Hz, 1H), 7.35-7.34 (m, 2H), , J = 1.0 Hz, 1H), 6.58 (s, 1H), 5.14 (bs, 1H), 4.54 (t, J = 7.5 Hz, 2H), 4.43-4.40 (m, 4H), 4.23-4.11 (m, 3H), 3.99-3.96 (m, 1H), .84 (m, 1H), 3.59 (s, 3H), 3.43-3.39 (m, 1H), 2.77-2.76 (m, 2H), 2.62-2.57 (m, 2H), 2.47-2.46 (m, 2H), 1.94-1.89 (m, 6H), 1.79-1.78 (m, 2H),1.69-1.66 (m, 2H) . 5 Example 244a 2-(4-Chloro(hydroxymethyl)pyridinyl)-3,4,6,7,8,9- hexahydropyrido[3,4-b]indolizin-1(2H)-one 244a To a solution of 4-chloro(1-oxo-3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin- yl)nicotinaldehyde 139a (1.0 g, 3.0 mmol) in methanol (30 mL) was added sodium 10 borohydride (380 mg, 9.0 mmol) at 30oC. The reaction mixture was d for another 1 h and quenched with water (10 mL). It was then concentrated under reduced pressure and the residue was extracted with dichloromethane (3 X 20 mL). The ed organic t was dried over anhydrous Na2SO4, filtered, and evaporated under reduced pressure to afford 244a as a yellow solid (920 mg, 92%). MS-ESI: [M+H]+ 332.3 15 Example 244b (4-Chloro(1-oxo-3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin- 2(1H)-yl)pyridinyl)methyl acetate 244b To a mixture of 244a (900 mg, 2.7 mmol) and triethylamine (810 mg, 8.1 mmol) in dichloromethane (30 mL) was added acetyl chloride (630 mg, 8.1 mmol) dropwise. The reaction mixture was stirred at 30oC for 1 h. It was then concentrated under reduced pressure 20 and the residue was purified by silica-gel column chromatography eluting with dichloromethane to afford 244b as white solid (900 mg, 90%). MS-ESI: [M+H]+ 374.2 349 Example 244c 3-(Acetoxymethyl)(1-oxo-3,4,6,7,8,9-hexahydropyrido[3,4- b]indolizin-2(1H)-yl)pyridinylboronic Acid 244c A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 244b (900 mg, 2.4 mmol), Pin2B2 (3.05 g, 12 mmol), 5 Pd(dppf)Cl2 (98 mg, 0.12 mmol), X-phos (114 mg, 0.24 mmol), potassium e (720 mg, 7.2 mmol), and dioxane (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at 65ºC for 4 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was washed with 5:1 eum ethyl acetate (10 mL) to afford 244c as a yellow solid (1.0 g, purity: 60%). MS-ESI: 10 [M+H]+ 384.1.
Example 244d (4-(1-Methyl(5-(1-methylazetidinyl)pyridinylamino)oxo- 1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin-2(1H)- yl)pyridinyl)methyl Acetate 244d A 50-mL round-bottomed flask equipped with a reflux condenser was d with 5- 15 bromomethyl(5-(1-methylazetidinyl)pyridinylamino)pyridin-2(1H)-one 239c (140 mg, 0.40 mmol), 244c (230 mg, 0.60 mmol), Pd(dppf)Cl2 (20 mg, 0.020 mmol), K3PO4 (180 mg, 0.80 mmol), sodium acetate·3water (120 mg, 0.80 mmol), water (0.5 mL), and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 2 h. It was then filtered and the filtrate was evaporated under reduced pressure. The 20 residue was purified on silica-gel column chromatography eluting with 25:1 dichloromethane/methanol to afford 244d as a yellow solid (90 mg, 43%). MS-ESI: [M+H]+ 608.3 Example 244 2-[3-(hydroxymethyl)[1-methyl[[5-(1-methylazetidinyl) pyridyl]amino]oxopyridyl]pyridyl]-3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin 25 one 244 A e of 244d (90 mg, 0.15 mmol) and lithium hydroxide (60 mg, 1.5 mmol) in propanol (5:3, 8 mL) and water (2 mL) was stirred at 30oC for 1 h. The e was evaporated in vacuo and the residue was diluted with water (3 mL). It was then extracted with ethyl acetate (20 mL X 2). The combined ethyl acetate extract was concentrated under 30 reduced pressure and the residue was ed by reverse-phase prep-HPLC to afford 244 (25 mg, 30 %) as white solid. MS-ESI: [M+H]+ 566.4. 1H NMR (500 MHz, CDCl3) δ 8.74 (d, J = 2.5 Hz, 1H), 8.51 (d, J = 5.5 Hz, 1H), 8.16 (d, J = 2.0 Hz, 1H), 7.91 (s, 1H), 7.87 (d, J = 2.0 Hz, 1H), 7.59-7.57 (m, 1H), 7.35 (d, J = 5.0 Hz, 1H), 6.85-6.83 (m, 1H), 6.33 (s, 1H), 5.04- 5.01 (m, 1H), 4.67-4.65 (m, 1H), 4.44-4.39 (m, 1H), 4.33-4.29 (m, 1H), 3.95-3.91 (m, 1H), 350 3.86-3.83 (m, 2H), 3.76-3.74 (m, 1H), 3.73 (s, 3H), 3.62-3.59 (m, 1H), 3.16-3.13 (m, 2H), 3.02-2.95 (m, 2H), 2.84-2.83 (m, 2H), 2.39 (s, 3H), 2.05-2.02 (m, 2H), 1.90-1.87 (m, 2H).
Example 245a 4-Chloro(1-oxo-6,7,8,9-tetrahydropyrido[3,4-b]indolizin-2(1H)- yl)nicotinaldehyde 245a 5 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 6,7,8,9-tetrahydropyrido[3,4-b]indolizin-1(2H)-one 112d (1.48 g, 7.9 mmol, 1.0 eq.), 2-bromochloronicotinaldehyde (3.48 g, 15.8 mmol, 2.0 eq.), 10 CuI (1.50 g, 7.9 mmol, 1.0 eq.), methoxy-1,10-phenanthroline (2.13 g, 7.9 mmol, 1.0 eq.), K2CO3 (2.18 g, 15.8 mmol, 2.0 eq.) and dioxane (50 mL). The reaction mixture was stirred at 100ºC for 24 h. After the reaction was completed, the mixture was filtered and the filtrate was concentrated under reduced pressure. The e was purified by silica-gel column chromatography eluting with 1:2 ethyl e/petroleum ether to afford 245a (550 15 mg, 21%) as a slight yellow solid. MS-ESI: [M+H]+ 328.1.
Example 245b 4-Fluoro(1-methyl(5-(4-(oxetanyl)piperazinyl)pyridin ylamino)-6 -oxo-1,6-dihydropyridinyl)(1-oxo-5,6,7,8-tetrahydro-1H-pyrrolo[3,4- b]indolizin-2(3H)-yl)benzaldehyde 245b A 50-mL -neck round-bottomed flask equipped with a reflux condenser was 20 charged with 245a (140 mg, 0.42 mmol, 1.0 eq.), (S)methyl(5-(2-methyl(oxetan yl)piperazinyl)pyridinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin- 2(1H)-one 191i (308 mg, 0.63 mmol, 1.5 eq.), Pd(dppf)Cl2 (35 mg, 0.042 mmol, 0.1 eq.), sodium acetate (70 mg, 0.84 mmol, 2.0 eq.), K3PO4 (175 mg, 0.84 mmol, 2.0 eq.), acetonitrile (20 mL), and water (1 mL). After three cycles of vacuum/argon flush, the mixture was heated 25 at 90oC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting e was purified by silica-gel 351 column chromatography eluting with 40:1 dichloromethane/methanol to afford 245b (100 mg, 36%) as a yellow solid. MS-ESI: [M+H]+ 647.4.
Example 245 (S)(3-(Hydroxymethyl)(1-methyl(5-(2-methyl(oxetanyl) piperazinyl)pyridinylamino)oxo-1,6-dihydropyridinyl)pyridinyl)-6,7,8,9- 5 tetrahydropyrido[3,4-b]indolizin-1(2H)-one 245 A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with 245b (100 mg, 0.15 mmol, 1.0 eq.), NaBH4 (29 mg, 0.77 mmol, 5.0 eq.), and methanol (10 mL). The resulting mixture was d at room temperature for 20 min. It was then filtered and the filtrate was concentrated under reduced pressure. The residue was 10 purified by reverse-phase prep-HPLC to afford 245 (80 mg, 79%) as a white solid. MS-ESI: [M+H]+ 649.3. 1H NMR (500 MHz, DMSO-d6) δ .68 (m, 1H), 8.57 (d, J = 5.0 Hz, 1H), 8.46 (s, 1H), 7.85 (d, J = 3.0 Hz, 1H), 7.55 (d, J = 2.5 Hz, 1H), 7.52 (d, J = 5.0 Hz, 1H), 7.39-7.36 (m, 1H), 7.26-7.24 (m, 2H), 6.69 (d, J = 7.5 Hz, 1H), 6.31 (s, 1H), 4.97 (bs, 1H), 4.58-4.54 (m, 2H), 4.49-4.41 (m, 2H), 4.34-4.27 (m, 2H), 4.09-4.06 (m, 2H), 3.69-3.68 (m, 15 1H), 3.61 (s, 3H), 3.41-3.39 (m, 1H), .19 (m, 1H), .93 (m, 1H), 2.87 (t, J = 6.0 Hz, 2H), 2.56-2.54 (m, 1H), 2.37-2.30 (m, 2H), 2.21-2.16 (m, 1H), 2.03-1.98 (m, 2H), 1.85- 1.82 (m, 2H), 0.94 (d, J = 5.5 Hz, 3H).
Example 246a 4-(5-(1,5-Dimethyl-1H-pyrazolylamino)methyloxo- 1,6-dihydropyridinyl)(1-oxo-6,7,8,9-tetrahydropyrido[3,4-b]indolizin-2(1H)- 20 yl)nicotinaldehyde 246a A 50-mL single-neck round-bottomed flask ed with a magnetic stirrer and a reflux condenser was charged with 4-chloro(1-oxo-6,7,8,9-tetrahydropyrido[3,4- lizin- 2(1H)-yl)nicotinaldehyde 245a (130 mg, 0.39 mmol, 1.0 eq.), 3-(1,5-dimethyl- 25 1H-pyrazolylamino)methyl(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin- 2(1H)-one 218a (600 mg, 1.75 mmol, 4.0 eq.), Pd(dppf)Cl2 (32 mg, 0.040 mmol, 0.1 eq.), sodium acetate (64 mg, 0.78 mmol, 2.0 eq.), K3PO4 (165 mg, 0.78 mmol, 2.0 eq.), acetonitrile (15 mL), and water (1 mL). After three cycles of vacuum/argon flush, the mixture was heated 352 at 90ºC for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 50:1 dichloromethane/ethanol to afford 246a (38 mg, 19%) as a yellow solid. MS-ESI: [M+H]+ 510.3. 5 Example 246 2-[4-[5-[(1,5-dimethylpyrazolyl)amino]methyloxo pyridyl](hydroxymethyl)pyridyl]-6,7,8,9-tetrahydropyrido[3,4-b]indolizinone 246 A 25-mL -neck round-bottomed flask equipped with a magnetic stirrer was charged with 246a (38 mg, 0.074 mmol, 1.0 eq.), NaBH4 (29 mg, 0.37 mmol, 5.0 eq.), and methanol (5 mL). The resulting mixture was stirred at room temperature for 20 min. It was 10 then filtered and the filtrate was trated. The residue was purified by e-phase prep-HPLC to afford the title compound (18 mg, 48%) as a white solid. MS-ESI: [M+H]+ 512.3. 1H NMR (500 MHz, CDCl 3) δ 8.58 (d, J = 5.0 Hz, 1H), 7.98 (d, J = 2.5 Hz, 1H), 7.78 (d, J = 2.5 Hz, 1H), 7.52 (d, J = 5.0 Hz, 1H), 7.37 (s, 1H), 7.17 (d, J = 7.0 Hz, 1H), 6.58-6.57 (m, 2H), 5.74 (s, 1H), 5.41-5.39 (m, 1H), 4.42-4.32 (m, 2H), 4.08 (t, J = 6.5 Hz, 2H), 3.72 (s, 15 3H), 3.70 (s, 3H), 2.98 (t, J = 6.5 Hz, 2H), 2.25 (s, 3H), 2.13-2.09 (m, 2H), 1.97-1.92 (m, 2H).
Example 247a {4-[5-({5-Acetyl-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin yl}amino)methyl- 6-oxo-1,6-dihydropyridinyl]{6-oxothia-4,5- ricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridinyl}methyl Acetate 247a 20 A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 3-(5-acetyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin ylamino)bromomethylpyridin-2(1H)-one 209c (183 mg, 0.50 mmol), {3- oxy)methyl]{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trien yl}pyridinyl}boronic acid 230i (200 mg, 0.50 mmol), f)Cl2 (37 mg, 0.05 mmol), 25 K3PO4 (212 mg, 1.0 mmol), sodium acetate (82 mg, 1.0 mmol), water (0.5 mL), and acetonitrile (5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 90oC for 1 h. It was then cooled to room temperature and filtered. The filtrate was concentrated 353 under reduced pressure and the resulting residue was ed by silica-gel column chromatography eluting with dichloromethane/methanol (50:1 to 20:1) to afford 247a as yellow solid (100 mg, 31 %). MS-ESI: [M+H]+ 641.2.
Example 247 5-[(5-acetyl-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazinyl)amino]- 5 1-methyloxopyridyl](hydroxymethyl)pyridyl]-6,7,8,9- tetrahydrobenzothiopheno[2,3-d]pyridazinone 247 A e of {4-[5-({5-acetyl-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazinyl}amino) methyloxo-1,6-dihydropyridinyl]{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca- (7),3-trienyl}pyridinyl}methyl acetate 247a(100 mg, 0.16 mmol) and lithium 10 hydroxide (96 mg, 4.0 mmol) in i-propanol/THF/water (2:2:1,10 mL) was stirred at 35oC for 30 min. The mixture was concentrated under reduced pressure. To the residue was added water (5 mL) and the resulting mixture was extracted with dichloromethane three times. The combined organic layer was then concentrated under reduced pressure and the resulting residue was purified by reverse-phase prep-HPLC to afford 247 (51 mg, 53%). MS-ESI: 15 [M+H]+ 599.3. 1H NMR (500 MHz, DMSO-d6, T=80oC) δ 8.54-8.51 (m, 1H), 8.38 (d, J = 3.0 Hz, 1H), 7.93-7.91 (m, 2H), 7.48 (d, J = 8.5 Hz, 1H), 7.35 (d, J = 3.0 Hz, 1H), 6.00 (s, 1H), 4.64-4.62 (m, 2H), 4.39 (s, 2H), 3.98-3.95 (m, 2H), 3.89-3.86 (m, 2H), 3.58 (s, 3H), 2.95- 2.93 (m, 2H), 2.87-2.84 (m, 2H), 2.08 (s, 3H), 1.89-1.87 (m, 4H).
Example 248a (R)-(6-Aminopyridinyl)(3-methylmorpholino)methanone 20 248a To a on of (R)methylmorpholine (2.02 g, 20 mmol) in ethanol (25 mL) was added 1-ethyl(3-dimethylaminopropyl) carbodiimide (EDCI) (3.33 g, 17.4 mmol), hydroxybenzotriazole (HOBt) (2.35 g, 17.4 mmol), and 6-aminonicotinic acid (2.0 g, 14.5 mmol). After stirring for 18 h at room temperature, the reaction sion was filtered and 25 the te was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 3:1 ethyl acetate/petroleum ether to afford 248a as white solid (1.6 g, 36%). MS-ESI: [M+H]+ 222.3. 354 Example 248b 6-Chloromethyl[(5-{[(3R)methylmorpholin yl]carbonyl}pyridinyl)amino]-2,3-dihydropyridazinone 248b A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 5 reflux condenser was charged with 1,4-dioxane (50 mL), 248a (330 mg, 1.5 mmol), o- 6-chloromethylpyridazin-3(2H)-one (446 mg, 2.0 mmol), cesium carbonate (978 mg, 3.0 mmol), Xantphos (88 mg, 0.15 mmol), and tris(dibenzylideneacetone)dipalladium(0) (68 mg, 0.075 mmol). The system was subjected to three cycles of vacuum/argon flush and heated at reflux for 4 h. It was then cooled to room temperature and filtered. The solid was washed 10 with dichloromethane (3 X 30 mL) and the combined filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with eum ether/ethyl acetate (2:1 to 1:2) to afford 248b (430 mg, 79%) as yellow solid. MSESI : [M+H]+ 364.3 e 248c (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodecadienyl}{1-methyl[(5-{[(3R)methylmorpholinyl]carbonyl}pyridin yl)amino]oxo-1,6-dihydropyridazinyl}pyridinyl)methyl Acetate 248c A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 248b (364 mg, 1.0 mmol), {3-[(acetoxy)methyl]{4,4- dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic 20 acid 199e (596 mg, 1.5 mmol), K3PO4 (424 mg, 2.0 mmol), 1,1’- bis(diphenylphosphino)ferrocenedichloropalladium(II) (73 mg, 0.10 mmol), sodium acetate (164 mg, 2.0 mmol), acetonitrile (10 mL), and water (0.5 mL). After three cycles of 355 vacuum/argon flush, the on mixture was heated at 100 °C for 2.5 h. Analysis of the reaction mixture by LCMS showed complete conversion to the desired product. The reaction e was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue was diluted with dichloromethane (50 mL) and water (50 mL). 5 The aqueous layer was separated and extracted with dichloromethane (3 × 20 mL). The combined organic layer was dried over Na2SO4 and filtered. The te was concentrated under reduced pressure and the dark residue was purified by silica-gel column tography eluting with dichloromethane/methanol (80:1 to 50:1) to afford 248c (250 mg, 37%) as yellow oil. MS-ESI: [M+H] + 681.3 10 Example 248 3-[3-(hydroxymethyl)[1-methyl[[5-[(3R)methylmorpholine carbonyl]pyridyl]amino]oxo-pyridazinyl]pyridyl]-7,7-dimethyl-1,2,6,8- ydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 248 To a solution of 248c (250 mg, 0.37 mmol) in THF/i-propanol/water(2.5/2/0.5 mL) was added lithium hydroxide (86 mg, 3.6 mmol) at room temperature. After the reaction was 15 stirred for 3 h, LCMS indicated the reaction was complete. Then the mixture was poured into water (20 mL) and extracted with dichloromethane (3 X 20 mL). The combined organic layer was washed with brine (30 mL), dried over Na2SO4, filtered. The filtrate was concentrated under reduced pressure. The residue solid was purified by reverse-phase prep-HPLC to afford 248 (110 mg, 48%) as a white solid. MS-ESI: [M+H] + 639.3. 1H NMR (500 MHz, MeOD) δ 20 8.85 (s, 1H), 8.58 (d, J = 5.0 Hz, 1H), 8.44 (d, J = 2.0 Hz, 1H), 7.79 (dd, J = 2.5, 8.5 Hz, 1H), 7.57 (d, J = 5.0 Hz, 1H), 7.32 (d, J = 8.5 Hz, 1H), 6.74 (s, 1H), .83 (m, 1H), 4.66-4.64 (m, 1H), 4.42-4.27 (m, 4H), 4.02-3.88 (m, over lap, 6H), 3.74-3.67 (m, 2H), 3.56-3.46 (m, 2H), 2.67-2.59 (m, 2H), 2.51 (s, 2H), 1.39 (d, J = 6.5 Hz, 3H), 1.28 (s, 6H).
Example 249a (R)Bromomethyl(5-(3-methylmorpholinecarbonyl)pyridin- 25 2-ylamino) pyridin-2(1H)-one 249a A 100-mL single-neck round-bottomed flask equipped with a ic stirrer and a reflux condenser was charged with 1,4-dioxane (15 mL), (R)-(6-aminopyridinyl) (3- 356 methylmorpholino)methanone 248a (332 mg, 1.5 mmol), 3,5-dibromomethylpyridin- 2(1H)-one (480 mg, 1.8 mmol), and cesium carbonate (978 mg, 3.0 mmol). After bubbling nitrogen through the suspension for 3 s, Xantphos (87 mg, 0.15 mmol) and tris(dibenzylideneacetone)dipalladium(0) (69 mg, 0.075 mmol) were added. The system was 5 subjected to three cycles of vacuum/argon flush and heated at reflux for 2.5 h. It was then cooled to room temperature and filtered. The solid was washed with dichloromethane (2 X 50 mL) and the combined filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with petroleum ether/ethyl e (2:1 to 1:2) to afford 249a (430 mg, 70%) as a yellow solid. MS-ESI: [M+H]+ 407.3 10 Example 249b 4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{1-methyl[(5-{[(3R)methylmorpholinyl]carbonyl}pyridin yl)amino]oxo-1,6-dihydropyridinyl}pyridinyl)methyl Acetate 249b A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with 249a (407 mg, 1.0 mmol), {3-[(acetyloxy)methyl]{4,4- 15 dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (800 mg, 2.0 mmol), K3PO4 (424 mg, 2.0 mmol), 1,1’- phenylphosphino)ferrocenedichloropalladium(II) (73 mg, 0.1 mmol), sodium acetate (164 mg, 2.0 mmol), acetonitrile (8 mL), and water (0.2 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 1.5 h. Analysis of the reaction 20 mixture by LCMS showed complete conversion to the desired product. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was d with dichloromethane (20 mL) and water (20 mL). The aqueous layer was separated and extracted with dichloromethane (20 mL × 3). The combined organic layer was dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The dark residue 25 was ed by silica-gel column chromatography g with 60:1 dichloromethane/methanol to afford 249b (200 mg, 29%) as yellow solid. MS-ESI: [M+H]+ 680.1 Example 249 3-[3-(hydroxymethyl)[1-methyl[[5-[(3R)methylmorpholine yl]pyridyl]amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- 30 tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 249 To a solution of 249b (204 mg, 0.30 mmol) in THF/i-propanol/water(3/3/0.5 mL) was added lithium hydroxide (72 mg, 3 mmol) at room temperature. After the reaction was stirred for 3 h, LCMS indicated the reaction was complete. The mixture was trated under reduced pressure and the residue was diluted with water (10 mL). It was then extracted with 357 dichloromethane (3 X 10 mL). The combined organic layer was washed with brine (30 mL), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase prep-HPLC (A: 1% NH4HCO3 in water, B: acetonitrile) to afford 249 (85 mg, 44%) as a white solid. MS-ESI: [M+H]+ 638.3. 1H NMR 5 (500 MHz, MeOD) δ 8.93 (d, J = 2.0 Hz, 1H), 8.52 (d, J = 5.0 Hz, 1H), 8.33 (d, J = 2.0 Hz, 1H), 7.69 (dd, J = 2.0, 6.5 Hz, 1H), 7.61 (d, J = 2.0 Hz, 1H), 7.48 (d, J = 5.0 Hz, 1H), 7.13 (d, J = 8.5 Hz, 1H), 6.74 (s, 1H), 4.70 (d, J = 12.0 Hz, 1H), 4.58 (d, J = 12.0 Hz, 1H), 4.40-4.27 (m, 4H), 3.99-3.89 (m, 3H), 3.74 (S, 3H), 3.55-3.46 (m, overlap, 4H), 2.67-2.59 (m, 2H), 2.51 (s, 2H), 1.39 (d, J = 6.5 Hz, 3H), 1.28 (s, 6H). 10 Example 250a 1-(5-(Hydroxymethyl)nitro-1H-pyrazolyl)propanol 250a A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with (3-nitro-1H-pyrazolyl)methanol (0.57 g, 4.0 mmol), Cs2CO3 (261 mg, 0.8 15 mmol), and yloxirane (20 mL). The mixture was stirred at 30oC for 2 days. The mixture was cooled to room temperature and diluted with dichloromethane (100 mL). The resulting mixture was filtered and the te was concentrated under d pressure. The residue was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 250a (0.40 g, 50%) as a white solid. : [M+H]+ 20 202.3 Example 250b 1-(5-(Bromomethyl)nitro-1H-pyrazolyl)propanol 250b To a mixture of 250a (4.0 g, 20.0 mmol) in chloroform (100 mL) cooled at 0 oC was added the solution of POBr3 (22.9 g, 80 mmol) in form (20 mL) over 30 s while maintaining the internal temperature below 5 oC. The reaction mixture was warmed to 50 oC 358 and stirred at this temperature for 3 h. It was then cooled to 0oC and quenched with water.
The organic layer was separated and evaporated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 250b (3.3 g, 62%) as yellow solid. MS-ESI: [M+H]+ 264.1 5 Example 250c 1-(5-((Methylamino)methyl)nitro-1H-pyrazolyl)propan- 2-ol 250c To a solution of 250b (3.0 g, 11.4 mmol) in romethane (30 mL) was added the solution of CH3NH2 (3.0 g, 34.2 mmol, 35% in water). This reaction mixture was d at room ature for 1 h. Then the organic layer was separated, dried over Na2SO4, and 10 filtered. The filtrate was concentrated under reduced pressure to afford 250c (1.9 g, 78%) as yellow solid. MS-ESI: [M+H]+ 215.3 Example 250d 5,6-Dimethylnitro-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine 250d To a mixture of 250c (1.129 g, 5.27 mmol) and triphenylphosphine (4.14 g, 15.8 15 mmol) in anhydrous THF (40 mL) cooled at 0oC was added the solution of di-isopropyl arboxylate (DIAD) (3.19 g, 15.8 mmol) in THF (15 mL) over a period of 30 minutes while maintaining the internal temperature below 5oC. The reaction mixture was warmed to 30oC and stirred at this temperature for 5 h. The mixture was then quenched with water (50 mL) and concentrated under reduced pressure. The residue was extracted with 20 dichloromethane (3 X 80 mL). The combined organic layer was dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was ed by silica-gel column chromatography eluting with dichloromethane/methanol (80:1 to 30:1) to afford 250d (0.83 g, 80%) as yellow solid. MS-ESI: [M+H]+ 197.2 Example 250e 5,6-Dimethyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin 25 amine 250e A solution of 250d (550 mg, 2.8 mmol) in methanol (20 mL) was added Raney Ni (about 600 mg). The reaction was charged with hydrogen gas (via balloon) and stirred for 2 h at room ature. It was then filtered through a plug of CELITE® and the filtrate was concentrated under d pressure to afford 250e as a yellow solid (400 mg, 86%), which 30 was used directly in the next step without further purification. : [M+H]+ 167.3 Example 250f (4-(5-(5,6-Dimethyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin- 2-ylamino)methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- dropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl Acetate 250f 359 A 50-mL round-bottomed flask ed with a magnetic stirrer and a reflux condenser was d with (4-(5-bromomethyloxo-1,6-di-hydropyridinyl)(1- oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl acetate 217a (525 mg, 1.0 mmol), 250e (166 mg, 1.0 mmol), cesium carbonate (652 mg, 2.0 mmol), and 5 1,4-dioxane (10 mL). After bubbling nitrogen h the suspension for 30 minutes, xantphos (116 mg, 0.20 mmol) and tris(dibenzylideneacetone)dipalladium(0) (92 mg, 0.10 mmol) were added. The system was subjected to three cycles of vacuum/argon flush and heated at reflux for 5 h. It was then cooled to room temperature and filtered. The solid was washed with dichloromethane (2 X 30 mL) and the combined filtrate was concentrated under 10 reduced pressure. The residue was purified by silica-gel column tography eluting with dichloromethane/methanol (80:1 to 30:1) to afford 250f (80 mg, 13%) as yellow solid. MSESI : [M+H]+ 611.4 Example 250 2-[4-[5-[(5,6-dimethyl-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazin yl)amino]methyloxopyridyl](hydroxymethyl)pyridyl]-3,4,6,7,8,9- 15 hexahydropyrazino[1,2-a]indolone 250 To a solution of 250f (75 mg, 0.123 mmol) in THF/i-propanol/water(4/2/2 mL) was added lithium hydroxide (15 mg, 0.62 mmol). The mixture was stirred at 30 oC for 1 h. After the reaction was complete, the mixture was evaporated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 250 as a white solid (40 mg, 20 57%). MS-ESI: [M+H]+ 569.3. 1H NMR (500 MHz, CDCl3) δ 8.49 (d, J = 5.0 Hz, 1H), 7.96 (bs, 1H), 7.73 (d, J = 2.0 Hz, 1H), 7.43 (s, 1H), 7.36 (d, J = 5.0 Hz, 1H), 6.91 (s, 1H), 5.71 (s, 1H), 5.03 (t, J = 3.5 Hz, 1H), 4.65-4.62 (m, 1H), 4.52-4.50 (m, 1H), 4.35-4.33 (m, 1H), 4.17- 4.05 (m, 3H), 3.91-3.88 (m, 2H), 3.73-3.71 (m, 1H), 3.71 (s, 3H), .52 (m, 1H), 2.90- 2.87 (m, 1H), 2.64-2.58 (m, 4H), 2.43 (s, 3H), 1.93-1.90 (m, 2H), 1.81-1.80 (m, 2H), 1.24 (d, 25 J = 6.5 Hz, 3H).
Example 252a o(1-ethylmethyl-1H-pyrazolylamino) methylpyridin-2(1H)-one 252a 360 A 100-mL round-bottomed flask was d with omethyl(5-methyl- 1H-pyrazolylamino)pyridin-2(1H)-one 115a (800 mg, 2.83 mmol), bromoethane (216 mg, 1.98 mmol), K2CO3 (780 mg, 5.66 mmol), and DMF (20 mL). The mixture was heated at 85oC overnight. It was then filtered and the filtrate was evaporated in vacuo. The e was 5 purified by -gel column chromatography g with 1:20 methanol/dichloromethane to afford 252a as a red solid (298 mg, 37%). MS-ESI: [M+H]+ 311.0. 1H NMR (500 MHz, DMSO-d6) δ 8.28 (s, 1H), 7.99 (d, J = 2.5 Hz, 1H), 7.35 (d, J = 2.5 Hz, 1H), 5.85 (s, 1H), 3.98-3.94 (m, 2H), 3.48 (s, 3H), 2.19 (s, 3H), 1.27 (t, J = 7.0 Hz, 3H).
Example 252b (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 10 2(6),7-dienyl}{5-[(1-ethylmethyl-1H-pyrazolyl)amino]methyloxo-1,6- dihydropyridinyl}pyridinyl)methyl Acetate 252b A round-bottomed flask equipped with a reflux condenser was charged with 252a (200 mg, 0.64 mmol), (3-(acetoxymethyl)(7,7-dimethyloxo-3,4,7,8-tetrahydro-1H- cyclopenta[4,5]pyrrolo[1,2-a]pyrazin-2(6H)-yl)pyridinyl)boronic acid 199e (309 mg, 0.64 15 mmol), PdCl2(dppf) (52.5 mg, 0.060 mmol), K3PO4 (333 mg, 1.29 mmol), sodium acetate (105 mg, 1.29 mmol), acetonitrile (10 mL), and water (0.5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 3 h. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified by silica-gel column chromatography eluting with 1:20 methanol/dichloromethane to afford 252b as a yellow solid 20 (120 mg, 27.6%). MS-ESI: [M+H]+ 584.3 Example 252 3-[4-[5-[(1-ethylmethyl-pyrazolyl)amino]methyloxo pyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 252 A mixture of 252b (120 mg, 0.21 mmol) and lithium hydroxide (23 mg, 0.82 mmol) 25 in THF (6 mL), i-propanol (4 mL), and water (2 mL) was stirred at room temperature for 0.5 h. It was then concentrated under reduced pressure and the residue was diluted with water (5 mL). The resulting mixture was ted with dichloromethane (2 X 10 mL). The combined dichloromethane extract was concentrated under reduced pressure. The residue was ed with reverse-phase prep-HPLC to afford 252 (52 mg, 47%) as a white solid. MS-ESI: 30 [M+H]+ 542.3. 1H NMR (500 MHz, DMSO-d6) δ 9.06 (d, J=5.5 Hz, 1H), 8.09 (s, 1H), 8.07 (s, 1 H), 7.41 (s, 1H), 7.33 (d, J=5.0 Hz, 1H), 6.56 (s, 1H), 5.88 (s, 1H), 4.97 (t, J = 4.5 Hz, 1H), 4.50-4.41 (m, 2H), .19 (m, 3H), 3.93-3.85 (m, 3H), 3.59 (s, 3 H), 2.62-2.57 (m, 2H), 2.43 (s, 2H), 2.19 (s, 3 H), 1.27-1.23 (m, overlap, 9 H). 361 Example 253a 4-Bromochloro((2- (trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one 253a A 500-mL single-neck bottomed flask equipped with a magnetic stirrer was 5 purged with nitrogen and charged with anhydrous DMF (150 mL) and 4-bromochloropyridazin-3 (2H)-one (10.0 g, 47.8 mmol). The reaction mixture was cooled to 0 ºC and sodium hydride was added. The reaction was d at 0 oC for 20 min. After this time, 2- (trimethylsilyl)ethoxymethyl chloride (11.9 g, 71.6 mmol) was added and the cooling bath was removed, and the reaction was stirred at room temperature for 3 h. The reaction was 10 then quenched with ted s sodium onate (30 mL). The mixture was ted with ethyl acetate (2 × 300 mL). The extracts were dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography to afford 253a in a 56% yield (9.00 g) as a yellow oil: 1H NMR (300 MHz, CDCl3) δ 8.02 (s, 1H), 5.42 (s, 2H), 3.79 (t, 2H, J = 5.4 Hz), 0.96 (t, 2H, J = 5.4 Hz), 0.01 (s, 15 9H).
Example 253b (S)-tert-Butyl 4-(6-(6-Chlorooxo((2- (trimethylsilyl)ethoxy)-methyl)-2,3-dihydropyridazinylamino)pyridinyl) methylpiperazinecarboxylate 253b A 50-mL round-bottomed flask equipped with a reflux condenser was charged with 20 (S)-tert-butyl 4-(6-aminopyridinyl)methylpiperazinecarboxylate 191f (580 mg, 2.0 mmol), 253a (1.36 g, 4.0 mmol), Pd2(dba)3 (180 mg, 0.20 mmol), Xantphos (230 mg, 0.40 mmol), Cs2CO3 (1.3 g, 4.0 mmol), and dioxane (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100 oC for 2 h. It was then cooled to room temperature and filtered. The filtrate was evaporated under reduced pressure and the resulting residue was 362 purified by silica-gel column chromatography eluting with 4:1 petroleum ether/ethyl acetate to afford 253b (1.0 g, 91%) as yellow solid. MS-ESI: [M+H]+ 551.2 Example 253c (S)Chloro(5-(2-methylpiperazinyl)pyridin ylamino)pyridazin-3(2H)-one 253c 5 A 50-mL round-bottomed flask was charged with 253b (551 mg, 1.0 mmol), concentrated HCl (2 mL), and methanol (10 mL). The mixture was stirred at room temperature overnight. It was then concentrated under d pressure to afford 253c, which was used directly in the next step t further purification. MS-ESI: [M+H]+ 321.1 Example 253d (S)Chloro(5-(2-methyl(oxetanyl)piperazin 10 yl)pyridinylamino)pyridazin-3(2H)-one 253d A 50-mL round-bottomed flask equipped with a magnetic stirrer was charged with 253c (321 mg, 1.0 mmol), 3-oxetanone (142 mg, 2.0 mmol), NaBH3CN (125 mg, 2.0 mmol), ZnCl2 (272 mg, 2.0 mmol), and methanol (10 mL). The mixture was stirred at room temperature overnight and concentrated under reduced pressure. Water (20 mL) was added to 15 the residue and the resulting mixture was ted with dichloromethane (3 X 20 mL). The combined organic layer was concentrated under reduced re and the residue was purified by -gel column chromatography eluting with ethyl acetate to afford 253d (210 mg, 56%) as yellow solid. : [M+H]+ 377.3.
Example 253e 4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 20 2(6),7-dienyl}[5-({5-[(2S)methyl(oxetanyl)piperazinyl]pyridin yl}amino)oxo-1,6- dihydropyridazinyl]pyridinyl)methyl Acetate 253e A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 253d (172 mg, 0.46 mmol), (3-(acetoxymethyl)(7,7- dimethyloxo-3,4,7,8-tetrahydro-1H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazin-2(6H)- 25 yl)pyridinyl)boronic acid 199e (0.91 g, 2.29 mmol), Pd(dppf)Cl2 (36 mg, 0.050 mmol), K3PO4 (195 mg, 0.92 mmol), sodium acetate (75 mg, 0.050 mmol), water (0.5 mL), and itrile (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at 90oC for 2 h. It was then cooled to room temperature and filtered. The filtrate was evaporated under reduced re and the resulting e was purified by silica-gel column 30 chromatography eluting with 20:1 ethyl acetate/methanol to afford the 253e (100 mg, 31%) as brown solid. MS-ESI: [M+H]+ 694.3.
Example 253 3-[3-(hydroxymethyl)[5-[[5-[(2S)methyl(oxetan yl)piperazinyl]pyridyl]amino]oxo-1H-pyridazinyl]pyridyl]-7,7-dimethyl- 1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 253 363 A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with 253e (92 mg, 0.13 mmol), lithium hydroxide (16 mg, 0.65 mmol), THF (2 mL), i-propanol (2 mL), and water (0.5 mL). The mixture was stirred at room temperature for 1 h.
It was then concentrated under reduced re. Water (10 mL) was added to the residue and 5 the resulting mixture was ted with dichloromethane (3 X 20 mL). The combined organic layer was concentrated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 253 (22 mg, 26%) as white solid. MS-ESI: [M+H]+ 652.2. 1H NMR (500 MHz, CDCl 3) δ 10.83 (s, 1H), 8.61 (s, 1H), 8.58 (d, J = 5.0 Hz, 1H), 8.12 (s, 1H), 8.05 (d, J = 2.5 Hz, 1H), 7.45 (d, J = 5.0 Hz, 1H), 7.32 (dd, J = 3.0 Hz, 5.5 Hz, 1H), 10 7.00 (d, J = 3.5 Hz, 1H), 6.84 (s, 1H), 4.74-4.68 (m, 3H), 4.65-4.58 (m, 3H), 4.26-4.14 (m, 2H), 3.99-3.96 (m, 1H), 3.71-3.69 (m, 1H), 3.55-3.53 (m, 1H), 3.18-3.14 (m, 2H), 2.64-2.59 (m, 3H), 2.53-2.47 (m, 4H), 2.40-2.33 (m, 2H), 1.29 (s, 6H), 1.09 (d, J = 7.0 Hz, 3H). e 254a 4-[5-({5-Acetyl-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin yl}amino)methyloxo-1,6-dihydropyridinyl]{4,4-dimethyloxothia 15 azatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienyl}pyridinecarbaldehyde 254a A 50-mL single-neck bottomed flask equipped with a ic r and a reflux condenser was charged with 3-(5-acetyl-4,5,6,7-tetrahydropyrazolo [1,5-a]pyrazin yl-amino)methyl(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 209d 20 (344 mg, 0.83 mmol), 4-chloro{4,4-dimethyloxothia azatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienyl}pyridinecarbaldehyde 109a (202 mg, 0.56 mmol), Pd(dppf)Cl2 (20 mg, 0.028 mmol), K3PO4 (235 mg, 1.11 mmol), sodium acetate (91 mg, 1.11 mmol), water (0.5 mL), and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 1 h. Analysis of the reaction 25 mixture by LCMS showed complete sion to the desired product. The mixture was cooled down to room temperature and ed. The filtrate was concentrated under reduced pressure to afford 254a (400 mg, crude), which was directly used in next step without further purification. MS-ESI: [M+H]+ 612.3 364 Example 254 3-[4-[5-[(5-acetyl-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazinyl)amino]- 1-methyloxopyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]thieno[1,3-c]pyridinone 254 To a solution of 254a (98 mg, 0.16 mmol) in methanol and dichloromethane was 5 added NaBH4 (13 mg, 0.33 mmol). The reaction mixture was stirred at room temperature for 1 h. is of the reaction mixture by LCMS showed complete conversion to the desired product. The mixture was quenched with aqueous NH4Cl solution (5 mL) and concentrated under reduced pressure. The residue was extracted with dichloromethane (3 X 10 mL). The combined extract was concentrated under reduced pressure and the residue was ed by 10 reverse-phase prep-HPLC to afford 254 (53 mg, 54%) as white solid. MS-ESI: [M+H]+ 613.9. 1H NMR (500 MHz, DMSO-d oC) δ 8.45 (d, J = 8.5 Hz, 1H), .92 (m, 2H), 7.33 6, T=80 (d, J = 3.5 Hz, 1H), 7.30 (d, J = 8.5 Hz, 1H), 5.97 (s, 1H), 4.67-4.63 (m, 3H), 4.46-4.45 (m, 2H), 3.97-3.93 (m, 2H), 3.89-3.86 (m, 3H), 3.56 (s, 3H), 2.97-2.91 (m, 2H), 2.53-2.55 (m, 2H), 2.49-2.46 (m, 2H), 2.08 (s, 3H), 1.21 (s, 6H). 15 Example 255a 4-(5-(5-Acetyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin ylamino)methyloxo-1,6-dihydropyridinyl)(10-fluorooxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 255a Following the ure described in Example 246, and starting with 4-chloro(10- 20 fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 134c (200 mg, 0.575 mmol) and 3-(5-acetyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinylamino) methyl(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 209d (356 mg, 0.863 mmol ), 255a was obtained as a red oil (320 mg, 93%). : [M+H]+599.3 Example 255 2-[4-[5-[(5-acetyl-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazinyl)amino]- 25 1-methyloxopyridyl](hydroxymethyl)pyridyl]fluoro-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indolone 255 Following the procedure described in Example 254, and ng with 255a(200 mg, 0.334 mmol), 255 was obtained as a white solid (55.5 mg, 28%). MS-ESI: [M+H]+ 601.3. 365 1HNMR (500 MHz, DMSO-d oC) δ 8.45 (d, J = 8.0 Hz, 1H), 7.93 (d, J = 3.5 Hz, 2H), 6, T=80 7.33 (d, J = 4.0 Hz, 1H), 7.30 (d, J = 8.5 Hz, 1H), 5.97 (s, 1H), 4.70-4.63 (m, 3H), 4.46 (d, J = 8.5 Hz, 2H), .86 (m, overlap, 8H), 3.58 (s, 3H), .55 (m, 2H), 2.43-2.39 (m, 2H), 2.08 (s, 3H), 1.79-1.67 (m, 4H). 5 e 256a (S)-(6-Aminopyridinyl)(3-methylmorpholino)methanone 256a To a solution of (S)methylmorpholine (1.5 g, 15.0 mmol) in ethanol (20 mL) was added EDCI (3.33 g, 17.4 mmol), HOBt (2.35 g, 17.4 mmol), and 6-aminonicotinic acid 10 (2.07 g, 15.0 mmol) at room temperature. After stirring for 18 h, the ing suspension was filtered. The solid was purified by silica-gel column chromatography eluting with 2:1 petroleum ether/ethyl acetate to straight ethyl acetate to afford 256a (1.0 g, 30%) as white solid. MS-ESI: 222.3 (M+H)+.
Example 256b (S)Bromomethyl(5-(3-methylmorpholinecarbonyl 15 pyridineylamino)pyridin-2(1H)-one 256b A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 1,4-dioxane (10 ml), 256a (111 mg, 0.50 mmol), 4- bromochloromethylpyridazin-3(2H)-one (134 mg, 0.60 mmol), cesium carbonate (326 mg, 1.0 mmol), Xantphos (29 mg, 0.05 mmol), and tris(dibenzylideneacetone)dipalladium(0) 20 (23 mg, 0.025 mmol). The system was subjected to three cycles of vacuum/argon flush and heated at 100oC for 5 h. It was then cooled to room temperature and filtered. The solid was washed with dichloromethane (3 X 30 mL) and the ed filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with petroleum ether/ethyl acetate (2:1 to 1:2) to afford 256b (140 mg, 77%) as a yellow solid. 25 : [M+H]+ 364.3 Example 256c (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{1-methyl[(5-{[(3S)methylmorpholinyl]carbonyl}pyridin yl)amino]oxo-1,6-dihydropyridazinyl}pyridinyl)methyl Acetate 256c 366 A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 256b (140 mg, 0.38 mmol), cetyloxy)methyl] {4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridin yl}boronic acid 199e (159 mg, 0.40 mmol), K3PO4 (85 mg, 0.40 mmol), sodium acetate (33 5 mg, 0.40 mmol), is(diphenylphosphino) ferrocenedichloropalladium(II) (15 mg, 0.020 mmol), acetonitrile (10 mL), and water (0.5 mL). The system was subjected to three cycles of vacuum/argon flush and heated at 100°C for 2.5 h. Analysis of the reaction mixture by LCMS showed te conversion to the desired product. The reaction mixture was cooled to room temperature and diluted with dichloromethane (30 mL) and water (30 mL). The s layer 10 was separated and extracted with dichloromethane (3 × 20 mL). The combined organic layer was dried over Na2SO4 and filtered. The te was concentrated under reduced re.
The dark residue was purified by silica-gel column chromatography eluting with 60:1 dichloromethane:/methanol to afford 256c (90 mg, 35%) as yellow solid. MS-ESI: [M+H]+ 681.3 15 Example 256 3-[3-(hydroxymethyl)[1-methyl[[5-[(3S)methylmorpholine yl]pyridyl]amino]oxo-pyridazinyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 256 To a solution of 256c (90 mg, 0.13 mmol) in THF/ anol /water(2.0/1/0.5 ml) was added lithium hydroxide (31 mg, 1.3 mmol) at room temperature. After the reaction was 20 stirred for 3 h, LCMS indicated the reaction was complete. Then the mixture was poured into water (15 mL) and extracted with dichloromethane (3 X 10 mL). The combined organic layer was washed with brine (30 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by reverse-phase prep- HPLC to afford 256 (40 mg, 48%) as white solid. MS-ESI: [M+H]+ 639.3. 1H NMR (500 25 MHz, MeOD) δ 8.86 (s, 1H), 8.58 (d, J = 5.0 Hz, 1H), 8.44 (d, J = 2.0 Hz, 1H), 7.79 (dd, J = 2.0, 6.5 Hz, 1H), 7.57 (d, J = 5.0 Hz, 1H), 7.32 (d, J = 8.5 Hz, 1H), 6.74 (s, 1H), 4.85-4.82 (m, 1H), 4.66-4.64 (m, 1H), 4.42-4.27 (m, 4H), 4.02-3.88 (m, overlap, 6H), 3.74-3.67 (m, 2H), 3.56-3.46 (m, 2H), 2.67-2.59 (m, 2H), 2.51 (s, 2H), 1.39 (d, J = 7.0 Hz, 3H), 1.28 (s, 6H).
Example 257 (S)Bromomethyl(5-(3-methylmorpholinecarbonyl)pyridine- 30 2-ylamino)pyridin-2(1H)-one 257a 367 A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with (S)-(6-aminopyridinyl)(3- methylmorpholino)methanone (222 mg, 1.0 mmol), 3,5-dibromomethylpyridin-2(1H)-one 5 (320 mg, 1.2 mmol), cesium carbonate (652 mg, 2 mmol), and 1,4-dioxane (10 mL). After bubbling nitrogen through the suspension for 10 minutes, Xantphos (58 mg, 0.10 mmol) and tris(dibenzylideneacetone)dipalladium(0) (46 mg, 0.050 mmol) were added. The system was t to three cycles of vacuum/argon flush and heated at reflux for 2.5 h. It was then cooled to room temperature and filtered. The solid was washed with dichloromethane (2 X 30 10 mL) and the combined te was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with petroleum ether/ethyl acetate (2:1 to 1:2) to afford 257a (280 mg, 69%) as a yellow solid. MS-ESI: [M+H]+ 407.3 Example 257b 4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{1-methyl[(5-{[(3S)methylmorpholinyl]carbonyl}pyridin 15 yl)amino]oxo-1,6-dihydropyridinyl}pyridinyl)methyl Acetate 257b A 25-mL single-neck round-bottomed flask equipped with a ic stirrer and a reflux condenser was charged with 257a (203 mg, 0.50 mmol), {3-[(acetoxy)methyl]{4,4- dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (640 mg, 1.6 mmol), K3PO4 (212 mg, 1.0 mmol), 1,1’- 20 bis(diphenylphosphino)ferrocenedichloropalladium(II) (18 mg, 0.025 mmol), sodium acetate (82 mg, 1.0 mmol), acetonitrile (10 mL), and water (0.2 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100°C for 2.5 h. Analysis of the on mixture by LCMS showed complete conversion to the desired product. The reaction mixture was cooled to room temperature and concentrated under reduced re. The residue was 25 diluted with dichloromethane (20 mL) and water (20 mL). The aqueous layer was separated and ted with dichloromethane (3 X 20 mL). The combined organic layer was dried over Na2SO4 and ed. The filtrate was concentrated under reduced pressure. The dark residue was purified by -gel column chromatography eluting with 60:1 dichloromethane /methanol to afford 257b (160 mg, 47%) as black solid. MS-ESI: [M+H]+ 680.1 368 Example 257 3-[3-(hydroxymethyl)[1-methyl[[5-[(3S)methylmorpholine carbonyl]pyridyl]amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 257 To a solution of 257b (157 mg, 0.23 mmol) in THF/i-propanol/water (2/2/0.5 mL) 5 was added lithium hydroxide (55 mg, 2.3 mmol) at room temperature. After the reaction was stirred for 3 h, LCMS indicated the reaction was completed. The e was poured into water (15 mL) and extracted with dichloromethane (3 X 15 mL). The combined organic layer was washed with brine (30 mL), dried over Na2SO4 and filtered. The filtrate was concentrated under d re. The residue solid was purified by reverse-phase prep-HPLC (A: 1‰ 10 3 in water; B: acetonitrile) to afford 257 (52 mg, 35%) as yellow solid. MS-ESI: [M+H]+ 668.3. 1H NMR (500 MHz, MeOD) δ 8.94 (d, J = 2.0 Hz, 1H), 8.53 (d, J = 5.0 Hz, 1H), 8.33 (d, J = 2.0 Hz, 1H), 7.69 (dd, J = 2.0, 6.5 Hz, 1H), 7.61 (d, J = 2.0 Hz, 1H), 7.48 (d, J = 5.0 Hz, 1H), 7.13 (d, J = 8.5 Hz, 1H), 6.74 (s, 1H), 4.70 (d, J = 12.0 Hz, 1H), 4.58 (d, J = 12.0 Hz, 1H), 4.41-4.27 (m, 4H), .89 (m, 3H), .66 (m, overlap, 5H), 3.55-3.46 15 (m, 2H), 2.67-2.59 (m, 2H), 2.51 (s, 2H), 1.39 (d, J = 6.5 Hz, 3H), 1.28 (s, 6H).
Example 258a 6-Chloromethyl({5-[(morpholinyl)carbonyl]pyridin yl}amino)-2,3-dihydropyridazinone 258a A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 20 reflux condenser was charged with 1,4-dioxane (40 mL), (6-aminopyridinyl)(morpholino)- methanone (2.07 g, 10.0 mmol), 4-bromochloromethylpyridazin-3(2H)-one 111a (3.35 g, 15.0 mmol), Pd2(dba)3 (915 mg, 1.0 mmol), XantPhos (578 mg, 1.0 mmol), and cesium carbonate (6.52 g, 20 mmol). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 8 h. It was then cooled to room temperature and filtered. The solid was 25 washed with dichloromethane (2 X 20 mL). The combined filtrate was dried over ous Na2SO4 and concentrated under reduced pressure to afford 258a (2.45 g, 51%) as a yellow solid. MS: [M+H]+ 350.1 Example 258b 1-Methyl(5-(morpholinecarbonyl)pyridinylamino) oxo-1,6- dihydropyridazinylboronic Acid 258b 369 A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 258a (2.0 g, 5.73 mmol, 1.0 eq.), 4,4,4',4',5,5,5',5'- octamethyl-2,2'-bi(1,3,2-dioxaborolane) (7.56 g, 28.6 mmol, 5.0 eq.), Pd(dppf)Cl2 (465 mg, 5 0.57 mmol, 0.1 eq.), X-Phos (461 mg, 1.14 mmol, 0.2 eq.), potassium acetate (1.11 g, 11.4 mmol, 2.0 eq.), and dioxane (50 mL). After three cycles of /argon flush, the mixture was heated at 50ºC for 6 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was washed with 1:3 ethyl acetate/petroleum ether to afford 258b (1.70 g, 83%) as a yellow solid, which was used in the 10 next step without further purification. MS: [M+H]+ 360.1 Example 258c 4-Chloro{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca- 1(9),2(7),3-trienyl} pyridinecarbaldehyde 258c A 100-mL round-bottomed flask equipped with a reflux condenser was charged with 4-chloro{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}pyridine- 15 3-carbaldehyde 124a (100 mg, 0.29 mmol), 258b (128 mg, 0.36 mmol), PdCl2(dppf) (24 mg, 0.031 mmol), K3PO4 (123 mg, 0.58 mmol), sodium acetate (57 mg, 0.58 mmol), acetonitrile (30 mL), and water (3 mL). After three cycles of vacuum/argon flush, the mixture was stirred at 100oC for 3 h. It was then filtered and the filtrate was evaporated in vacuo. The e was ed with silica-gel column tography eluting with 1:3 petroleum/ethyl e to 20 afford 258c as a yellow solid (45 mg, 25%). MS-ESI: [M+H]+ 625.2 Example 258 3-[3-(hydroxymethyl)[1-methyl[[5-(morpholinecarbonyl) pyridyl]amino]oxo-pyridazinyl]pyridyl]-6,7,8,9-tetrahydrobenzothiopheno[2,3- d]pyridazinone 258 A mixture of 258c (45 mg, 0.071 mmol), NaBH4 (8 mg, 0.21 mmol), and methanol (7 25 mL) was d at room temperature for 2 h. Then the reaction mixture was ed with water (5 mL) and concentrated under reduced pressure. The residue was extracted with dichloromethane (2 X 10 mL) and the combined dichloromethane extract was concentrated under reduced pressure. The residue was ed by reverse-phase prep-HPLC to afford 258 (24 mg, 53%) as a yellow solid. MS-ESI: [M+H]+ 627.2. 1H NMR (500 MHz, DMSO-d6) δ 370 9.80 (s, 1H), 8.68 (s, 1H), 8.65 (d, J = 4.5 Hz, 1H), 8.50 (s, 1H), 8.39 (d, J = 2.0 Hz, 1H), 7.80-7.78 (m, 1H), 7.67 (d, J = 5.0 Hz, 1H), 7.60 (d, J = 8.5 Hz, 1H), 4.85 (t, J = 5.5 Hz, 1H), 4.52-4.35 (m, 2H), 3.82 (s, 3H), 3.60-3.49 (m, 8H), 2.95-2.93 (m, 2H), 2.89-2.84 (m, 2H),1.94-1.83 (m, 4H). 5 Example 259a 2-{4,4-Dimethyloxothiaazatricyclo[6.4.0.02,6]dodeca- 1(8),2(6)-dienyl}[1-methyl({5-[(morpholinyl)carbonyl]pyridinyl}amino) oxopyridazinyl]pyridinecarbaldehyde 259a A round-bottomed flask equipped with a reflux condenser was charged with 4-chloro- 10 2-{4,4-dimethyloxothiaazatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienyl}pyridine- 3-carbaldehyde 109a (144 mg, 0.40 mmol), 1-methyl(5-(morpholinecarbonyl)pyridin- 2-ylamino)oxo-1,6-dihydropyridazinylboronic acid 258a (215 mg, 0.60 mmol), PdCl2(dppf) (16 mg, 0.020 mmol), K3PO4 trihydrate (207 mg, 0.80 mmol), sodium acetate (66 mg, 0.80 mmol), acetonitrile (15 mL), and water (1.5 mL). After three cycles of 15 /argon flush, the e was d at 100oC for 2 h. It was then filtered and the filtrate was evaporated in vacuo. The e was purified with -gel column chromatography eluting with 25:1 dichloromethane/methanol to afford 259a as a yellow solid (80 mg, 30%). MS-ESI: [M+H]+ 640.3.
Example 259 3-[3-(hydroxymethyl)[1-methyl[[5-(morpholinecarbonyl) 20 pyridyl]amino]oxo-pyridazinyl]pyridyl]-7,7-dimethyl-1,2,6,8- ydrocyclopenta[3,4]thieno[1,3-c]pyridinone 259 A mixture of 259a (80 mg, 0.12 mmol), NaBH4 (14 mg, 0.36 mmol), and methanol (5 mL ) was stirred at room temperature for 1 h. It was then quenched with brine (5 mL) and evaporated under reduced pressure. The residue was extracted with dichloromethane (2 x 10 25 mL) and the combined dichloromethane extract was concentrated under reduced pressure.
The resulting residue was purified by reverse-phase prep-HPLC to afford 259 as a white solid (38 mg, 49%). MS-ESI: [M+H]+ 642.8. 1H NMR (500 MHz, CDCl3) δ 8.78 (s, 1H), 8.61 (d, J = 5.0 Hz, 1H), 8.44 (d, J = 2.5 Hz, 2H), 7.80-7.77 (m, 1H), 7.45 (d, J = 5.0 Hz, 1H), 7.06- 7.04 (m, 1H), 4.65-4.59 (m, 2H), 4.42 (s, 1H), 4.30-4.27 (m, 1H), 3.95 (s, 3H), 3.90-3.87 (m, 371 1H), 3.76-3.68 (m, 8H), 3.04-2.92 (m, 2H), .80 (m, 2H), 2.59-2.54 (m, 2H), 1.30 (s, 6H).
Example 260a (2-{4,4-Dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{1-methyl[(5-methyl-1H-pyrazolyl)amino]oxo-1,6- 5 dihydropyridinyl}pyridin yl)methyl Acetate 260a A round-bottomed flask equipped with a reflux condenser was charged with 5-bromo- yl(5-methyl-1H-pyrazolylamino)pyridin-2(1H)-one 218a (201 mg, 0.71 mmol), {3-[(acetyloxy)methyl]{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- 10 dienyl}pyridinyl}boronic acid 199e (282 mg, 0.71 mmol), Pd(dppf)Cl2 (51 mg, 0.07 mmol), K3PO4 (301 mg, 1.42 mmol), sodium acetate (116 mg, 1.42 mmol), acetonitrile (10 mL), and water (0.2 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 3 h. It was then filtered and the filtrate was evaporated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 1:20 15 methanol/dichloromethane to afford 260a as a red solid (150 mg, 38%). MS-ESI: [M+H]+ 556.3 e 260 3-[3-(hydroxymethyl)[1-methyl[(5-methyl-1H-pyrazol yl)amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 260 20 A mixture of 260a (150 mg, 0.27 mmol) and lithium hydroxide (13 mg, 0.54 mmol) in THF (6 mL), i-propanol (4 mL), and water (2 mL) was stirred at room temperature for 0.5 h. The mixture was concentrated under reduced pressure and the e was diluted with water (5 mL). It was then extracted with dichloromethane (2 x 10 mL) and the combined dichloromethane extract was trated under reduced pressure. The residue was purified 25 with reverse-phase prep-HPLC to afford 260 (28 mg, 20%) as a white solid. MS-ESI: [M+H]+ 514.3. 1H NMR (500 MHz, DMSO-d6) δ 11.76 (s, 1 H), 8.47 (d, J = 5.0 Hz, 1 H), 8.07-8.05 (m, 2 H), 7.38-7.31 (m, 2 H), 6.55 (s, 1 H), 5.88 (s, 1 H), 4.95-4.93 (m, 1 H), 4.48- 372 4.39 (m, 2 H), 4.22-4.18 (m, 3 H), 3.83 (d, J = 5.5 Hz, 1 H), 3.58 (s, 3 H), 2.64-2.56 (m, 2 H), 2.36-2.34 (m, 2 H), 2.16 (s, 3 H), 1.22 (s, 6 H).
Example 261a 4-{5-[(1,5-Dimethyl-1H-pyrazolyl)amino]methyloxo- 1,6-dihydro nyl}{10-fluorooxo-1H,2H,3H,4H,6H,7H,8H,9H-pyrido[3,4- 5 b]indolizinyl}pyridinecarbaldehyde 261a A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 4-chloro(10-fluorooxo-3,4,6,7,8,9-hexahydropyrido [3,4-b]indolizin-2(1H)-yl)nicotinaldehyde 134c (97 mg, 0.28 mmol), 3-[(1,5-dimethyl-1H- 10 pyrazolyl)amino]methyl(tetramethyl-1,3,2-dioxaborolanyl)-1,2-dihydropyridin one 218a (192.6 mg, 0.56 mmol), Pd2(dba)3 (54.9 mg, 0.060 mmol), tri(cyclohexyl)phsphine (50.2 mg, 0.18 mmol), Cs2CO3 (182.6 mg, 0.56 mmol), dioxane(8 mL), and water (0.25 mL).
After three cycles of vacuum/argon flush, the mixture was d at 110oC for 2 h. It was then filtered and the filtrate was evaporated in vacuo. The residue was purified by silica-gel 15 column tography eluting with 35:1 ethyl e/methanol to afford 261a (90 mg, 61%) as a black solid. MS-ESI: [M+H]+ 530.2 Example 261 5-[(1,5-dimethylpyrazolyl)amino]methyloxopyridyl]- 3-(hydroxymethyl)pyridyl]fluoro-3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizinone 261 20 To a solution of 261a (90.0 mg, 0.17 mmol) in methanol (5 mL) was added sodium borohydride (64.6 mg, 1.7 mmol) at room temperature. The on was stirred for 0.5 h. It was then quenched with water (2 mL) and ated in vacuo. The residue was extracted with dichloromethane (3 X 10 mL). The combined dichloromethane extract was concentrated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 25 261 (47.0 mg, 52%) as a white solid. MS-ESI: [M+H]+ 532.3. 1H NMR (500 MHz, DMSO- d6) δ 8.47 (d, J = 5.0 Hz, 1H), 8.05 (s, 1H), 8.03 (d, J = 2.5 Hz, 1H), 7.39 (d, J = 2.5 Hz, 1H), 7.31(d, J = 5.0 Hz, 1H), 5.89 (s, 1H), 4.87-4.85 (m, 1H), 4.45-4.36 (m, 2H), 4.11-4.09 (m, 1H), 3.93-3.91 (m, 1H), 3.79-3.76 (m, 2H), 3.59 (s, 3H), 3.58 (s, 3H), 3.00-2.94 (m, 2H), 2.66-2.63 (m, 2H), 2.18 (s, 3H), 1.90-1.88 (m, 2H), 1.78-1.73 (m, 2H) 373 Example 262a omethyl(5-methyloxazolylamino)pyridin- 2(1H)-one 262a A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 5 reflux condenser was charged with 5-methyloxazolamine (276 mg, 2.82 mmol), 3,5- dibromomethylpyridin-2(1H)-one (753 mg, 2.82 mmol), tris- (dibenzylideneacetone)dipalladium(0) (256 mg, 0.28 mmol), os (324 mg, 0.56 mmol), Cs2CO3 (1.8 g, 5.64 mmol), and 1,4-dioxane (30 mL). After three cycles of vacuum/argon flush, the mixture was heated at 92oC for 3 hrs. It was then cooled to room temperature and 10 filtered. The filtrate was concentrated under reduced pressure and the resulting residue was ed by silica-gel column chromatography eluting with 100:1 dichloromethane/methanol to afford 262a as white solid (702 mg, 88 %). MS-ESI: [M+H]+ 284.1.
Example 262b (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl} {1-methyl[(5-methyl-1,3-oxazolyl)amino]oxo-1,6- 15 opyridinyl} pyridin yl)methyl e 262b A 50-mL single-neck bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 262a (150 mg, 0.53 mmol), cetyloxy)methyl] {4,4-dimethyloxo-1,10-diazatricyclo [6.4.0.02,6]dodeca-2(6),7-dienyl}pyridin yl}boronic acid 199e (421 mg, 1.06 mmol), Pd(dppf)Cl2 (37 mg, 0.050 mmol), K3PO4 (225 20 mg, 1.06 mmol), sodium acetate (87 mg, 1.06 mmol), water (0.5 mL), and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at 90oC for 1 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (50:1 to 30:1) to afford 262b as yellow solid (100 mg, 34%). 25 MS-ESI: [M+H]+ 556.9.
Example 262 3-[3-(hydroxymethyl)[1-methyl[(5-methyloxazolyl)amino] oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- b]pyrazinone 262 374 A mixture of 262b (100 mg, 0.18 mmol) and lithium hydroxide (108 mg, 4.5 mmol) in i-propanol/THF/water (4/4/2 mL) was stirred at 35oC for 30 min. The mixture was concentrated under reduced pressure. To the residue was added water (5 mL) and the resulting mixture was extracted with dichloromethane three times. The combined organic 5 layer was concentrated under reduced pressure and the resulting e was purified by e-phase PLC to afford 262 as white solid (21.0 mg, 23%). MS-ESI: [M+H]+ 515.3. 1H NMR (500 MHz, DMSO-d6) δ 9.19 (s, 1H), 8.49 (d, J = 5.0 Hz, 1H), 8.30 (s, 1H), 7.60 (d, J = 1.0 Hz, 1H), 7.32 (d, J = 4.5 Hz, 1H), 6.62 (s, 1H), 6.56 (s, 1H), 4.96-4.94 (m, 1H), 4.45-4.36 (m, 2H), 4.24-4.14 (m, 3H), 3.84 (d, J = 10.5 Hz, 1H), 3.59 (s, 3H), 2.62-2.56 10 (m, 2H), 2.44-2.42 (m, 2H), 2.22 (s, 3H), 1.22 (s, 6H).
Example 263a 6-Chloromethyl(pyrimidinylamino)pyridazin-3(2H)- one 263a N N O O N NH B B N NH O O O O N HO N Cl N f)Cl2 , X-phos, B N KOAc , dioxane, 50 oC, 6 h OH 263a 263b N N NH O N O 134a N Pd(dppf)Cl2, K3PO4 N N H2O, 80oC, 4h, dioxane F O N 263c A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 15 reflux condenser was charged with 1,4-dioxane (150 mL), pyrimidinamine (1.7 g, 18.0 mmol), 4-bromochloromethylpyridazin-3(2H)-one (4.0 g, 18.0 mmol), and cesium carbonate (11.74 g, 36.0 mmol). After ng nitrogen through the suspension for 30 minutes, Xantphos (1.04 g, 1.8 mmol) and tris(dibenzylideneacetone)dipalladium(0) (823 mg, 0.9 mmol) were added. The system was subjected to three cycles of vacuum/argon flush and 20 heated at reflux for 15 h. It was then cooled to room temperature and ed. The solid was washed with dichloromethane (2 X 50 mL). The combined filtrate was concentrated and the 375 residue was washed with acetonitrile (5 mL) to afford 263a (2.99 g, 70%) as a yellow solid.
MS: [M+H]+ 238 Example 263b 1-methyloxo(pyrimidinylamino)-1,6- dihydropyridazinyl-boronic acid 263b 5 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 263a (500 mg, 2.11 mmol), 4,4,4',4',5,5,5',5'-octamethyl- 2,2'-bi(1,3,2-dioxaborolane) (2.68 g, 10.6 mmol), Pd(dppf)Cl2 (170 mg, 0.20 mmol), X-phos (170 mg, 0.40 mmol), ium acetate (410 mg, 4.21 mmol), and dioxane (30 mL). The system was subjected to 3 cycles of vacuum/argon flush and stirred at 50oC for 6 h. LCMS 10 indicated that 263a was totally converted to 263b.
Example 263c 2-(10-Fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl)(1-methyloxo(pyrimidinylamino)-1,6-dihydropyridazin yl)nicotinaldehyde 263c To the mixture of 263b at room temperature was added 4-chloro(10-fluorooxo- 15 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 134c (300 mg, 0.90 mmol), Pd(dppf)Cl2 (170 mg, 0.20 mmol), K3PO4 (103 mg, 0.40 mmol), and water (2 mL).
The system was subjected to 3 cycles of vacuum/argon flush again and d at 80oC for 4 h.
The reaction mixture was then cooled to room temperature, diluted with water (30 mL), and filtered. The filtrate was extracted with dichloromethane (2 x 30 mL). The combined 20 dichloromethane t was concentrated under reduced pressure and the e was purified by silica-gel column chromatography eluting with romethane/methanol (40:1 to 20:1) to afford 263c as a yellow solid (210 mg, 45%). MS-ESI: [M+H]+ 515.3 Example 263 10-fluoro[3-(hydroxymethyl)[1-methyloxo(pyrimidin ylamino)pyridazinyl]pyridyl]-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indolone 263 25 A 100-mL -neck round-bottomed flask equipped with a magnetic stirrer was charged with 263c (100 mg, 0.19 mmol), NaBH4 (30 mg, 0.78 mmol), and methanol (20 mL).
The e was d at room temperature for 0.5 h. It was then diluted with water (30 mL) and concentrated under reduced pressure. The residue was extracted with dichloromethane (2 X 30 mL) and the combined dichloromethane extract was dried and concentrated under 30 reduced pressure. The e was purified with reverse-phase prep-HPLC to afford 263 as a white solid (67 mg, 68%). MS-ESI: [M+H]+ 517.3. 1H NMR (500 MHz, DMSO-d6) δ 9.93 (s, 1H), 8.81 (s, 1H), 8.67 (s, 1H), 8.55 (d, J = 4.5 Hz, 1H), 8.49 (d, J = 5.5 Hz, 1H), 7.56 (dd, J = 1.0, 6.0 Hz, 1H), 7.43 (d, J = 5.0 Hz, 1H), 4.83 (t, J = 5.5 Hz, 1H), 4.62-4.58 (m, 1H), 4.39- 376 4.36 (m, 1H), 4.25-4.19 (m, 2H), 4.06-4.04 (m, 1H), 3.92-3.90 (m, 1H), 3.81 (s, 3H), 2.64- 2.54 (m, 2H), 2.43-2.41 (m 2H), 1.781.76 (m, 2 H), 1.69-1.67 (m, 2H).
Example 264a 2-(4-Aminopyrimidinyl)propanol 264a Br O N N HO N Br N NH O N NH2 Pd2(dba)3, Xantphos OH N Cs2CO3, dioxane Br 100 °C, 15 h 264a 264b N HO N NH 199e OAc O N f)Cl2, K3PO4, THF N N H2O, reflux, 6h O N 264c 5 To a solution of ethyl 4-aminopyrimidinecarboxylate (840 mg, 5.0 mmol) in anhydrous tetrahydrofuran (50 mL) cooled at -20 oC was added a solution of methylmagnesium bromide in THF (8.5 mL, 25.0 mmol, 3.0 M) over a period of 5 minutes.
The reaction e was stirred at 0oC for another 2 h. It was then quenched with saturated NH4Cl (20 mL) and concentrated under reduced pressure. The residue was extracted with 10 ethyl acetate (5 X 40 mL). The combined organic layer was dried over anhydrous Mg2SO4, ed, and ated under reduced pressure. The residue was purified by reverse-phase Combiflash to afford 264a as yellow solid (240 mg, 32%) MS-ESI: [M+H]+ 154.1 Example 264b 5-Bromo(2-(2-hydroxypropanyl)pyrimidinylamino) methylpyridin-2(1H)-one 264b 15 A 100-mL single-neck bottomed flask equipped with a magnetic r and a reflux condenser was charged with 264a (300 mg, 2.0 mmol), 3,5-dibromomethylpyridin- 2(1H)-one (800 mg, 3.0 mmol), Pd2(dba)3 (182 mg, 0.20 mmol), XantPhos (231 mg, 0.40 mmol), Cs2CO3 (1.30 g, 4.0 mmol), and 1,4-dioxane (20 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 15 h. It was then cooled to room 20 temperature and filtered. The filtrate was concentrated under reduced pressure. The resulting residue was purified by -gel column chromatography eluting with 40:1 377 dichloromethane/methanol (40:1) and further ed by reverse-phase Combiflash to afford 264b as white solid (200 mg, 30%). MS-ESI: [M+H]+ 339.0 Example 264c 4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}(5-{[2-(2-hydroxypropanyl)pyrimidinyl]amino}methyloxo- 5 1,6-dihydro-pyridinyl)pyridinyl)methyl Acetate 264c A 50-mL round-bottomed flask equipped with a reflux condenser was d with 264b (170 mg, 0.50 mmol), etoxymethyl)(7,7-dimethyloxo-3,4,7,8-tetrahydro- 1H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazin-2(6H)-yl)pyridinyl)boronic acid 199e (200 mg, 0.50 mmol), Pd(dppf)Cl2 (40 mg, 0.050 mmol), K3PO4 (212 mg, 1.0 mmol), water (0.5 mL), 10 and tetrahydrofuran (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at reflux for 6 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 40:1 dichloromethane/methanol to afford 264c as brown solid (200 mg, 54%). MS-ESI: [M+H]+ 612.3 15 Example 264 3-[3-(hydroxymethyl)[5-[[2-(1-hydroxymethyl-ethyl)pyrimidin- mino]methyloxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 264 To a solution of 264c (170 mg, 0.27 mmol) in tetrahydrofuran (10 mL) and water (2 mL) was added lithium hydroxide (64 mg, 3.0 mmol). The reaction mixture was stirred at 20 35oC for 2 h. It was then concentrated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 264 (86 mg, 46%) as yellow solid. MS-ESI: [M+H]+ 570.1. 1H NMR (500 MHz, DMSO-d6) δ 9.27 (s, 1H), 8.94 (d, J = 2.0 Hz, 1H), 8.50 (d, J = 5.0 Hz, 1H), 8.33 (d, J = 6.0 Hz, 1H), 7.76 (d, J = 2.5 Hz, 1H), 7.41(d, J = 5.0 Hz, 1H), 7.22 (d, J = 5.5 Hz, 1H), 6.56 (s, 1H), 5.14 (t, J = 5.0 Hz, 1H), 4.89 (s, 1H), 4.48-4.42 (m, 2H), 25 4.23-4.19 (m, p, 3H), 3.85-3.84 (m, 1H), 3.62(s, 3H), 2.67-2.56 (m, 2H), 2.42 (s, 2H), 1.42 (s, 3H), 1.40 (s, 3H), 1.21 (s, overlap, 6H).
Example 265a 1-(2-Nitro-6,7-dihydropyrazolo[1,5-a]pyrazin-5(4H)-yl)propan- 1-one 265a 378 To a solution of 2-nitro-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine 209a (200 mg, 1.19 mmol) in dichloromethane (8 mL) was added Et3N (240 mg, 2.38 mmol). After stirring for 5 minutes, a solution of propionyl chloride (121 mg, 1.31 mmol) in dichloromethane (2 mL) 5 was added and the reaction mixture was stirred at room temperature for 1 h. Analysis of the reaction e by LCMS showed complete conversion to the desired t. The mixture was washed with water and brine, dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to afford 265a (260 mg, 98%) as white solid, which was used in the next step without further purification. MS-ESI: [M+H]+ 225.0 10 Example 265b 1-(2-Amino-6,7-dihydropyrazolo[1,5-a]pyrazin-5(4H)- yl)propanone 265b To a solution of 265a (260 mg, 1.16 mmol) in methanol (10 mL) was added 10% Pd/C (26 mg). The system was evacuated and then refilled with H2. After stirring for 2 h, analysis of the reaction e by LCMS showed complete conversion to the desired 15 product. The mixture was filtered, and the filtrate was trated under reduced re to afford 265b as a yellow solid (225 mg, 99%). MS-ESI: [M+H]+ 195.1 Example 265c 5-Bromomethyl(5-propionyl-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)pyridin-2(1H)-one 265c A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 20 reflux condenser was charged with 265b (200 mg, 1.03 mmol), bromo methylpyridin-2(1H)-one (414 mg, 1.55 mmol), Pd2(dba)3 (47 mg, 0.052 mmol), Xantphos 379 (60 mg, 0.103 mmol), Cs2CO3 (671.6 mg, 2.06 mmol), and dioxane (20 mL). After three cycles of vacuum/argon flush, the reaction mixture was heated at 100oC for 3 h. Analysis of the reaction mixture by LCMS showed complete conversion to the desired product. The reaction mixture was cooled to room ature and filtered. The filtrate was concentrated 5 under reduced pressure and the residue was purified by silica-gel column chromatography eluting with 80:1 dichloromethane/methanol to afford 265c (280 mg, 72%) as white solid.
MS-ESI: [M+H]+ 380.2 e 265d 4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}[1-methyloxo({5-propanoyl-4H,5H,6H,7H-pyrazolo[1,5- 10 a]pyrazinyl}amino)-1,6-dihydropyridinyl]pyridinyl)methyl Acetate 265d A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 265c (200 mg, 0.53 mmol), {3-[(acetoxy)methyl]{4,4- dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (834 mg, 2.10 mmol), f)Cl2 (19 mg, 0.0263 mmol), K3PO4 (223 mg, 1.052 15 mmol), sodium acetate (86 mg, 1.052 mmol), acetonitrile (10 mL), and water (5 drops). After three cycles of vacuum/argon flush, the reaction mixture was heated at 100 oC for 3 h.
Analysis of the reaction e by LCMS showed complete conversion to the desired product. The reaction e was cooled down to room temperature and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica-gel column 20 chromatography g with 60:1 dichloromethane/methanol to afford 265d (100 mg, 29%) as yellow oil. MS-ESI: [M+H]+ 653.3 Example 265 3-[3-(hydroxymethyl)[1-methyloxo[(5-propanoyl-6,7- o-4H-pyrazolo[1,5-a]pyrazinyl)amino]pyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 265 25 To a solution of 265d (100 mg, 0.153 mmol) in THF (3 mL), i-propanol (3 mL), and water (5 mL) was added lithium hydroxide (37 mg, 1.53 mmol). The reaction mixture was stirred at room temperature for 1 h. Analysis of the reaction mixture by LCMS showed complete conversion to the desired t. The mixture was concentrated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 265 (50 mg, 30 54%) as white solid. MS-ESI: [M+H]+ 611.3. 1H NMR (500 MHz, DMSO-d6, T=80oC) δ 8.45 (d, J = 8.5 Hz, 1H), 7.93-7.90 (m, 2H), 7.35 (d, J = 3.5 Hz, 1H), 7.29 (d, J = 8.5 Hz, 1H), 6.55 (s, 1H), 5.98 (s, 1H), 4.74-4.71 (m, 1H), 4.65 (s, 2H), 4.46-4.44 (m, 2H), 4.18-4.16 (m, 2H), 3.97-3.87 (m, p, 5H), 3.58 (s, 3H), 2.57-2.56 (m, 2H), 2.49-2.37 (m, 4H), 1.22 (s, 6H), 1.03 (t, J = 12.0 Hz, 3H). 380 Example 266a 5-bromomethyl((5-(oxetanyl)-1H-pyrazol yl)amino)pyridin-2(1H)-one 266a Following the reaction scheme of Figure 26, 266a was prepared. 5 Example 266b (2'-(7,7-dimethyloxo-3,4,7,8-tetrahydro-1H- cyclopenta[4,5]pyrrolo[1,2-a]pyrazin-2(6H)-yl)methyl((5-(oxetanyl)-1H-pyrazol yl)amino)oxo-1,6-dihydro-[3,4'-bipyridin]-3'-yl)methyl acetate 266b A 25-mL -neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 266a (33 mg, 0.10 mmol), {3-[(acetyloxy)methyl]{4,4- 10 dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (60 mg, 0.15 mmol), Pd(dppf)Cl2 (7 mg, 0.010 mmol), K3PO4 (42 mg, 0.20 mmol), sodium acetate (16 mg, 0.20 mmol), itrile (6 mL), water (0.1 mL). After three cycles of /argon flush, the mixture was heated at reflux for 2 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under d pressure and the 15 resulting residue was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 266b as a white solid (17 mg, 28%). MS-ESI: [M+H]+ 598.4 Example 266 3-[3-(hydroxymethyl)[1-methyl[[5-(oxetanyl)-1H- pyrazolyl]amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- 20 tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 266 A mixture of 266b (15 mg, 0.025 mmol) and lithium hydroxide (6 mg, 0.25 mmol) in i-propanol/THF (1:1, 4 mL) and water (1 mL) was stirred at 30oC for 1 h. The mixture was evaporated under reduced re and the residue was diluted with water (5 mL). It was then extracted with ethyl acetate (2 X 10 mL). The combined ethyl e extract was 25 concentrated under reduced pressure and the residue was purified by reverse-phase prep- HPLC to afford 266 (4.5 mg, 33%) as a white solid. MS-ESI: [M+H]+ 556.3. 1H NMR (500 MHz, CDCl3) δ 8.49 (d, J = 5.0 Hz, 1H), 7.98 (s, 1H), 7.68 (s, 1H), 7.58 (s, 1H), 7.33 (d, J = 5.0 Hz, 1H), 6.86 (s, 1H), 6.09 (s, 1H), 5.07-5.04 (m, 2H), 4.79-4.76 (m, 2H), 4.68-4.66 (m, 381 1H), 4.54-4.51 (m, 1H), 4.37-4.34 (m, 1H), 4.28-4.25 (m, 1H), 4.18 (d, J = 5.5 Hz, 2H), 3.89- 3.87 (m, 1H), 3.73 (s, 3H), 2.59-2.57 (m, 2H), 2.54-2.52 (m, 3H), 1.29 (s, 6H).
Example 267a 5-Bromomethyl(5-methyl-1,3,4-thiadiazolylamino)pyridin- 2(1H)-one 267a 5 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 5-methyl-1,3,4-thiadiazolamine (1.15 g, 10.0 mmol), 3,5-dibromomethylpyridin-2(1H)-one (4.00 g, 15.0 mmol), Pd2(dba)3 (916 mg, 1.0 mmol), Xantphos (1.16 g, 2.0 mmol), Cs2CO3 (6.52 g, 20.0 mmol), and dioxane (50 mL). After three 10 cycles of vacuum/argon flush, the mixture was heated at 100oC for 3 h. It was then cooled to room temperature and ed. The te was evaporated under reduced re and the resulting residue was purified by -gel column chromatography eluting with 20:1 dichloromethane/methanol to afford 267a (2.2 g, 73%) as white solid. MS-ESI: [M+H]+ 301.2 15 e 267b (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{1-methyl[(5-methyl-1,3,4-thiadiazolyl)amino]oxo-1,6- dihydropyridinyl}pyridinyl)methyl Acetate 267b A 50-mL round-bottomed flask equipped with a reflux condenser was charged with 267a (150 mg, 0.50 mmol), {3-[(acetoxy)methyl]{4,4-dimethyloxo-1,10- 20 diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (640 mg, 1.5 mmol), PdCl2(dppf) (37 mg, 0.050 mmol), K3PO4 (212 mg, 1.0 mmol), sodium acetate (82 mg, 1.0 mmol), acetonitrile (10 mL), and water (0.2 mL). After three cycles of vacuum/argon flush, the e was heated at 90oC for 2 h. It was then cooled to room temperature and filtered. The filtrate was ated under reduced pressure and the resulting 25 residue was purified by silica-gel column chromatography eluting with 50:1 dichloromethane/methanol to afford 267b (80 mg, 28%) as yellow solid. MS-ESI: [M+H]+ 574.2 Example 267 3-[3-(hydroxymethyl)[1-methyl[(5-methyl-1,3,4-thiadiazol yl)amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- 30 tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 267 382 A 25-mL round-bottomed flask equipped with a magnetic stirrer was charged 267b (80 mg, 0.14 mmol), lithium hydroxide (17 mg, 0.70 mmol), THF (2 mL), i-propanol (2 mL), and water (0.5 mL). The mixture was stirred at room temperature for 1 h and concentrated under reduced pressure. The residue was diluted with water (5 mL) extracted with 5 dichloromethane (10 mL × 3) and the combined organic layer was concentrated under reduced pressure. The e was purified by reverse-phase PLC to afford 267 (32 mg, 43%) as white solid. MS-ESI: [M+H]+ 532.3. 1H NMR (500 MHz, DMSO-d6) δ 10.19 (s, 1H), 8.57 (d, J = 3.0 Hz, 1H), 8.49 (d, J = 6.0 Hz, 1H), 7.63 (d, J = 3.0 Hz, 1H), 7.32 (d, J = 6.0 Hz, 1H), 6.56 (s, 1H), 4.92 (t, J = 6.5 Hz, 1H), .37 (m, 2H), 4.22-4.17 (m, 3H), 10 3.85-3.80 (m, 1H), 3.60 (s, 3H), 2.58-2.56 (m, 2H), 2.52 (s, 3H), 2.42 (s, 2H), 1.17 (s, 6H).
Example 268a 1-Methylnitro-1H-imidazole 268a To a mixture of 4-nitro-1H-imidazole (2.0 g, 17.7 mmol) and K2CO3 (3.67 g, 26.5 mmol) in acetonitrile (20 mL) was added iodomethane (1.3 mL, 3.0 g, 21.2 mmol) dropwise 15 while stirring at room temperature. The ing mixture was stirred at 60oC overnight. It was then ated under reduced pressure and the residue was diluted with water (20 mL).
The mixture was extracted with dichloromethane (2 X 20 mL). The ed extract was concentrated under reduced pressure and the residue was purified by silica-gel column chromatography eluting with 100:1 dichloromethane/methanol to afford 268a as a yellow 20 solid (1.8 g, 82%). MS-ESI: [M+H]+ 128.1. 1H NMR (500 MHz, DMSO-d6) δ 8.37 (s, 1H), 7.82 (s, 1H), 3.76 (s, 3H).
Example 268b 1-Methyl-1H-imidazolamine 268b 383 A 100-mL round-bottomed flask was charged with 268a (1.6 g, 12.6 mmol), 10% palladium on carbon (50% wet, 160 mg), and ethanol (15 mL). The flask was evacuated, charged with hydrogen gas, and stirred at room temperature overnight. The catalyst was removed by filtration through a pad of CELITE® and the te was concentrated under 5 reduced pressure to afford 268b (1.2 g, 98%) as a yellow solid. MS-ESI: [M+H]+ 98.2 e 268c 5-Bromomethyl(1-methyl-1H-imidazol ylamino)pyridin-2(1H)-one 268c A 100-mL single-neck round-bottomed flask ed with a magnetic r and a reflux condenser was charged with oxane (50 mL), 268b(1.1 g, 11.3 mmol), 3,5- 10 dibromomethylpyridin-2(1H)-one (3.0 g, 11.3 mmol), Pd2(dba)3 (1.0 g, 1.13 mmol), XantPhos (1.3 g, 2.26 mmol), and cesium carbonate (7.3 g, 22.6 mmol). After three cycles of vacuum/argon flush, the mixture was heated at 92oC for 4.5 hrs. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column tography eluting with 15 dichloromethane/methanol (100:1 to 50:1) to afford 268c (2.4 g, 76 %) as yellow solid. MSESI : [M+H]+ 283.1 Example 268d (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dien yl}{1-methyl[(1-methyl-1H-imidazolyl)amino]oxo-1,6- dihydropyridinyl}pyridinyl)methyl Acetate 268d 20 A 50-mL round-bottomed flask ed with a reflux condenser was charged with 268c (150 mg, 0.53 mmol), {3-[(acetyloxy)methyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (80.4 mg, 0.21 mmol), Pd(dppf)Cl2 (17.2 mg, 0.021 mmol), K3PO4 (89 mg, 0.42 mmol), sodium acetate (57.1 mg, 0.42 mmol), water (0.2 mL), and acetonitrile (10 mL). After three cycles of 25 vacuum/argon flush, the mixture was heated at 90oC for 2.5 hrs. It was then cooled to room ature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (30:1 to 20:1) to afford 268d (110 mg, 37.2%) as brown solid.
MS-ESI: [M+H]+ 556.4. 30 Example 268 3-[3-(hydroxymethyl)[1-methyl[(1-methylimidazolyl)amino]- 6-oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- b]pyrazinone 268 A mixture of 268d (100 mg, 0.18 mmol) and lithium hydroxide (189 mg, 4.5 mmol) in i-propanol/THF (1:1, 4.0 mL) and water (1.0 mL) was stirred at 35oC for 30 min. The 384 mixture was concentrated under reduced pressure. To the residue was added water (5 mL) and the ing mixture was extracted with dichloromethane (3 X 10 mL). The combined organic layer was concentrated under reduced re and the resulting residue was purified by reverse-phase PLC to afford 268 (19.8 mg, 22%) as a yellow solid. MS-ESI: 5 [M+H]+ 514.3. 1H NMR (500 MHz, DMSO-d6) δ 8.47 (d, J = 5.5 Hz, 1H), 7.57 (s, 1H), 7.44 (d, J = 2.5 Hz, 1H), 7.39 (s, 1H), 7.36 (d, J = 2.5 Hz, 1H), 7.34 (d, J = 5.0 Hz, 1H), 6.95 (s, 1H), 6.56 (s, 1H), 5.12-5.10 (m, 1H), 4.44-4.41 (m, 2H), 4.22-4.18 (m, 3H), 3.84-3.82 (m, 1H), 3.60 (s, 3H), 3.59 (s, 3H), 2.59-2.56 (m, 2H), .42 (m, 2H), 1.22 (s, 6H).
Example 269a 2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 10 2(6),7-dienyl}[1-methyl({5-[(morpholinyl)carbonyl]pyridinyl}amino)oxo- 1,6-dihydropyridinyl]pyridinecarbaldehyde 269a A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 4-chloro{4,4-dimethyloxo-1,10- 15 diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 108a (100 mg, 0.29 mmol), 1-methyl(5-(morpholinecarbonyl)pyridinylamino)(4,4,5,5- tetramethyl-1,3,2-di-oxaborolanyl)pyridin-2(1H)-one 111c (192 mg, 0.44 mmol), Pd(dppf)Cl2 (12 mg, 0.015 mmol), K3PO4 (123 mg, 0.58 mmol), sodium acetate (47 mg, 0.58 mmol), acetonitrile (10 mL), and water (5 drops). After three cycles of vacuum/N2 flush, the 20 mixture was heated at 100 oC for 1 h. Analysis of the reaction mixture by LCMS showed complete conversion to the desired product. It was cooled to room temperature, and filtered.
The filtrate was concentrated under reduced pressure and the residue was purified by silicagel column chromatography (dichloromethane/methanol 40:1) to afford 269a (150 mg, 83%) as a brown solid. MS-ESI: [M+H]+ 621.8 25 Example 269 hydroxymethyl)[1-methyl[[5-(morpholinecarbonyl) l]amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 269 385 To a solution of 269a (150 mg, 0.24 mmol) in dichloromethane (5 mL) and methanol (5 mL) was added NaBH4 (18.2 mg, 0.482 mmol). After stirring at room temperature for 1 h, the mixture was quenched with aqueous NH4Cl (10 mL) and concentrated under reduced pressure. The residue was ted with dichloromethane (3 X 20 m). The combined extract 5 was washed with brine, dried over Na2SO4, concentrated under reduced pressure, and purified by reverse-phase prep-HPLC to afford 269 (114 mg, 76%) as a white solid. MS-ESI: [M+H]+ 624.3. 1H NMR (500 MHz, 6) δ 9.00 (s, 1H), 8.79 (d, J = 2.0 Hz, 1H), 8.50 (d, J = 4.5 Hz, 1H), 8.26 (d, J = 1.5 Hz, 1H), 7.67 (dd, J = 2.0, 9.0 Hz, 1H), 7.61 (d, J = 1.5 Hz, 1H), 7.38-7.36 (m, 2H), 6.56 (s, 1H), 5.00 (s, 1H), 4.47-4.40 (m, 2H), 4.25-4.19 (m, 3H), 10 3.86-3.84 (m, 1H), 3.62-3.60 (overlap, m, 4H), 3.51 (s, 3H), 3.52-3.50 (m, 4H), 2.59-2.57 (m, 2H), 2.43 (s, 2H), 1.22 (s, 6H).
Example 270a Ethyl N-[(Pyrazinyl)carbamothioyl]carbamate 270a A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer was 15 charged with pyrazinamine (7.6 g, 80.0 mmol, 1.0 eq.), O-ethyl carbonisothiocyanatidate (12.5 g, 95.4 mmol, 1.2 eq.), and dioxane (150 mL). The on mixture was stirred at room temperature for 24 hours. After the reaction was complete, it was trated to a volume of around 20 mL under reduced pressure and the resulting suspension was ed. The solid was collected and washed with ethyl acetate (3 × 20 mL) to afford 270a (14.0 g, 77%) as a 20 white solid. MS-ESI:[M+H]+ 227.3 Example 270b [1,2,4]Triazolo[1,5-a]pyrazinamine 270b A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 270a (6.00 g, 26.43 mmol, 1.0 eq.), hydroxylamine hydrochloride (3.32 g, 47.52 mmol, 1.8 eq.), DIPEA(12 mL), ethanol (40 mL), and methanol 386 (40 mL). The reaction mixture was d at 65oC for 16 hours. After the reaction was te, it was cooled to room temperature and concentrated to a volume of around 20 mL under d pressure. The resulting suspension was collected by filtration and the solid was washed with 60:1 dichloromethane/ethanol (50 mL) to afford 270b (3.3 g, 92%) as a white 5 solid. MS-ESI:[M+H]+ 136.3. 1H NMR (500 MHz, DMSO-d6) δ 8.84 (d, J = 1.0 Hz, 1H), 8.70 (dd, J = 1.0, 4.0 Hz, 1H), 7.98 (d, J = 5.0 Hz, 1H), 6.47 (s, 2H).
Example 270c tert-Butyl [1,2,4]Triazolo[1,5-a]pyrazinylcarbamate 270c A 250-mL single-neck round-bottomed flask equipped with a ic stirrer was charged with 270b (2.00 g, 14.8 mmol, 1.0 eq.), Boc2O (3.87 g, 17.77 mmol, 1.2 eq.), and 10 anhydrous THF (60 mL). The system was evacuated and refilled with N2. The reaction mixture was cooled to -78 ºC, followed by the addition of LHMDS (37.0 mL, 37.0 mmol, 2.5 eq., 1.0M in THF). After the reaction was stirred at -78OC for 2 hours, it was quenched with saturated aqueous NH4Cl solution (30 mL). The mixture was concentrated under reduced pressure and the residue extracted with dichloromethane (3 X 50 mL). The combined organic 15 layer was dried over sodium sulfate, filtered, and concentrated under reduced pressure. The resulting e was purified by silica-gel column chromatography eluting with 1:4 ethyl e/petroleum ether to afford 270c (1.87 g, 53%) as a white solid. MS-ESI:[M-t-Bu]+ 180.0. 1H NMR (500 MHz, DMSO-d6) δ 10.31 (s, 1H), 8.94 (s, 1H), 8.72 (d, J = 3.5 Hz, 1H), 7.95 (d, J = 4.0 Hz, 1H), 1.25 (s, 9H). 20 e 270d tert-Butyl 5,6,7,8-Tetrahydro-[1,2,4]triazolo[1,5-a]pyrazin ylcarbamate 270d A 100-mL round-bottomed flask was purged with nitrogen and charged with 270c (1.0 g, 4.25 mmol), 20% palladium on carbon (10% wet, 200 mg), and ethanol (40 mL). It was then evacuated, charged with hydrogen gas (25 atm), and stirred at 60oC for 24 h. The 25 hydrogen was evacuated and nitrogen was charged into the flask. The catalyst was removed by filtration through a pad of CELITE® and the filtrate was concentrated under reduced pressure to afford 270d (700 mg, 68%). MS-ESI: u]+184.0 Example 270e tert-Butyl 7-Methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5- a]pyrazinyl- carbamate 270e 30 Following the procedure in Example 191i, and starting with 270d (500 mg, 2.1 mmol, 1.0 eq.), paraformaldehyde (630 mg, 21.0 mmol, 10.0 eq.), diethyl ether (2.1 mL, 2.1 mmol, 1.0 M), NaBH3CN (390 mg, 6.3 mmol, 3.0 eq.), and methanol (20 mL) afforded 270e as a yellow solid (500 mg, 94%). MS-ESI: [M-tBu]+198.0. 387 Example 270f 7-Methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazin amine 270f Following the procedure in Example 131e, and starting with 270e (500 mg, 1.97 mmol) Boc deprotection with acid afforded 270f as a yellow solid (200 mg, 66%). MS-ESI: 5 [M+H]+ 154.1.
Example 270g (4-(1-Methyl(7-methyl-5,6,7,8-tetrahydro- [1,2,4]triazolo[1,5-a]pyrazinylamino)oxo-1,6-dihydropyridinyl)(1-oxo- 3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl Acetate 270g A 50-mL single-neck round-bottomed flask equipped with a ic stirrer and a 10 reflux condenser was charged with 270f (100 mg, 0.65 mmol, 1.7 eq.), (4-(5-bromo methyloxo-1,6- dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl)pyridinyl)methyl acetate 217a (200 mg, 0.38 mmol, 1.0 eq.), DMF (10 mL), and cesium carbonate (499 mg, 1.52 mmol, 4.0 eq.). After bubbling nitrogen h the resulting solution for 10 minutes, Xantphos (44 mg, 0.076 mmol, 0.20 eq.) and 15 tris(dibenzylideneacetone)dipalladium(0) (35 mg, 0.038 mmol, 0.10 eq.) were added. The reaction mixture was subjected to three cycles of vacuum/argon flush and heated at 100 oC for 16 h. After this time the reaction was cooled to room temperature and filtered. The filtrate was partitioned between ethyl acetate (50 mL) and water (10 mL). The aqueous layer was separated and extracted with ethyl acetate (3 × 20 mL). The combined organic layer was 20 concentrated under d pressure. The residue was purified on -gel column chromatography eluting with 50:1 dichloromethane/methanol to afford 270g (90 mg, 41%).
MS-ESI: [M+H]+ 598.3.
Example 270 2-[3-(hydroxymethyl)[1-methyl[(7-methyl-6,8-dihydro-5H- [1,2,4]triazolo[1,5-a]pyrazinyl)amino]oxopyridyl]pyridyl]-3,4,6,7,8,9- 25 hexahydropyrazino[1,2-a]indolone 270 Following the procedure for Example 241, and ng with 270g(90 mg, 0.15 mmol), afforded 270 as a white solid (47 mg, 56%). MS-ESI: [M+H]+ 556.3. 1H NMR (500 MHz, DMSO-d6) δ 8.50 (d, J = 5.0 Hz, 1H), 8.03 (d, J = 2.5 Hz, 1H), 7.83 (s, 1H), 7.50 (d, J = 2.5 Hz, 1H), 7.33 (d, J = 5.5 Hz, 1H), 6.58 (s, 1H), 4.93 (t, J = 5.5 Hz, 1H), 4.44-4.38 (m, 2H), 30 4.24-4.02 (m, 5H), .85 (m, 1H), 3.61-3.59 (m, p, 5H), 2.87 (t, J = 5.5 Hz, 2H), 2.64-2.58 (m, 2H), 2.49-2.46 (m, 2H), 2.40 (s, 3H), 1.80-1.69 (m, 4H). e 271a (S)-tert-Butyl 4-(6-(5-Chloromethoxypyridin ylamino)pyridinyl)methylpiperazinecarboxylate 271a 388 Boc N Boc N N N 1 N NH N NH 9 OH O 9 N O e 1, Pd2(dba)3, , N N N Cl Cs2CO3, dioxane, H2O, O N 110oC, 16 h 271a 2, LiOH, 45oC, 4h 271b HN N e/HCl N NH HO O 50oC, 2 h N N NH O N 271c A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 1,4-dioxane (40 mL), (S)-tert-butyl 4-(6-amino pyridin yl)methylpiperazinecarboxylate 101h (2.04 g, 7.0 mmol), 3-bromochloro 5 methoxypyridine (2.8 g, 12.6 mmol), Pd2(dba)3 (640 mg, 0.70 mmol), XantPhos (404.6 mg, 0.70 mmol), and cesium carbonate (4.56 g, 14.0 mmol). After three cycles of vacuum/argon flush, the mixture was heated at 100 oC for 4 h. After this time the reaction was cooled to room temperature. It was then filtered and the filtrate was evaporated under reduced pressure.
The residue was purified by silica-gel column chromatography eluting with 1:3 ethyl 10 acetate/petroleum ether to afford 271a (1.7 g, 57%) as a yellow solid. MS-ESI: [M+H]+ 434.2 Example 271b tert-Butyl (3S)(6-{[5-(2-{4,4-Dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}(hydroxymethyl)pyridinyl) ypyridinyl] pyridinyl)methylpiperazinecarboxylate 271b A 100-mL single-neck round-bottomed flask ed with a magnetic stirrer and a 15 reflux condenser was charged with 271a (650 mg, 1.50 mmol), {3-[(acetyloxy)methyl] {4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- dienyl}pyridin yl}boronic acid 199e (1.79 g, 4.5 mmol), Pd2(dba)3 (137.2 mg, 0.15 mmol), P(cy)3(167.4 mg, 0.60 mmol), Cs2CO3 (978 mg, 3.0 mmol), dioxane (20 mL), and water (0.5 mL). After three cycles of /argon flush, the mixture was heated at 110oC for 16 h. After this time the 389 reaction was cooled to room ature. Lithium ide monohydrate (1.89 g, 45 mmol) and water (2.0 mL) were added. The resulting e was stirred at 45oC for 4 h. It was then filtered and the te was evaporated under reduced pressure. The residue was purified by silica-gel column chromatography g with 3:1 ethyl acetate/petroleum ether to afford 5 271b (290 mg, 27%) as a yellow solid. MS-ESI: [M+H]+ 709.3 Example 271c 10-[3-(Hydroxymethyl)[5-({5-[(2S)methylpiperazin yl]pyridinyl}amino)oxo-1,6-dihydropyridinyl]pyridinyl]-4,4-dimethyl-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienone 271c A solution of 271b (286.6 mg, 0.40 mmol) in dioxane/HCl (30 mL) was stirred at 50 10 oC for 2 h. It was evaporated under reduced pressure to afford 271c (450 mg, crude) as a black solid. MS-ESI: [M+H]+ 595.3 Example 271 3-[3-(hydroxymethyl)[5-[[5-[(2S)methyl(oxetan yl)piperazinyl]pyridyl]amino]oxo-1H-pyridinyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 271 15 To a solution of 271c (450 mg, 0.75 mmol) in ol (10 mL) was added oxetan one (162 mg, 2.25 mmol), NaBH3CN (141.8 mg, 2.25 mmol), and ZnCl2 (306 mg, 2.25 mmol). The reaction was stirred at room temperature for 3 h. The mixture was evaporated under reduced pressure and the residue was diluted with water (5 mL). It was then extracted with dichloromethane (3 X 10 mL) and the combined dichloromethane extract was 20 concentrated under reduced pressure. The residue was purified by e-phase prep-HPLC to afford 271 (23.0 mg, 8.8%, over two steps) as a yellow solid. MS-ESI: [M+H]+651.3. 1H NMR (500 MHz, CDCl3) δ 9.76 (s, 1H), 8.74 (d, J = 2.0 Hz, 1H), 8.53 (d, J = 5.0 Hz, 1H), 7.99 (d, J = 3.0 Hz, 1H), 7.84 (s, 1H), 7.73 (s, 1H), 7.41 (d, J = 4.5 Hz, 1H), 7.35 (dd, J = 2.5 Hz, 8.5 Hz, 1H), 6.87 (s, 1H), 6.85 (d, J = 9.0 Hz, 1H), 5.16-5.13 (m, 1H), 4.72-4.69 (m, 5H), 25 4.54-4.53 (m, 1H), 4.36-4.35 (m, 1H), 4.19-4.17 (m, 2H), 3.89-3.87 (m, 1H), 3.56-3.49 (m, 2H), 3.11-3.09 (m, 2H), 2.60-2.48 (m, overlap, 7H), 2.24-2.21 (m, 1H), 1.29 (s, 6H), 1.02 (d, J = 6.0 Hz, 3H) Example 272a 5-Bromomethyl(5-(1-(oxetanyl)azetidinyl)pyridin- 2-ylamino)pyridine-2(1H)-one 272a 390 A mixture of 3-(5-(azetidinyl)pyridinylamino)bromomethylpyridin-2(1H)- one 239b (140 mg, 0.42 mmol), one (91 mg, 1.26 mmol), NaBH3CN (78 mg, 1.26 mmol), and zinc chloride (171 mg, 1.26 mmol) in methanol (10 mL) was stirred at 50oC for 2 5 hours. The mixture was trated under reduced pressure and water (5 mL) was added to the residue. It was then extracted with dichloromethane (3 X 10 mL). The combined organic layer was concentrated under reduced pressure. The e was purified by column chromatography eluting with 50:1 dichloromethane/methanol to afford 272a (145 mg, 85%).
MS-ESI: [M+H]+ 390.8 10 Example 272b 4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dien yl}[1-methyl({5-[1-(oxetanyl)azetidinyl]pyridinyl}amino) oxopyridinyl]pyridinyl)methyl Acetate 272b A 25-ml round-bottomed flask equipped with a reflux condenser was charged with 272a (140 mg, 0.35 mmol), {3-[(acetoxy)methyl]{4,4-dimethyloxo-1,10-diaza- 15 tricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (140 mg, 0.35 mmol), Pd(dppf)Cl2 (28 mg, 0.035 mmol), sodium acetate (58 mg, 0.70 mmol,), K3PO4 (148 mg, 0.70 mmol), water (6 drops), and acetonitrile (6 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 2 h. It was then filtered and the filtrate was ated under reduced pressure. The e was purified by silica-gel column 20 chromatography eluting with 25:1 dichloromethane/methanol to afford 272b (114 mg, 46%) as a brown solid. MS-ESI: [M+H]+ 664.4 Example 272 3-[3-(hydroxymethyl)[1-methyl[[5-[1-(oxetanyl)azetidin yl]pyridyl]amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 272 25 A mixture of 272b (114 mg, 0.17 mmol) and lithium hydroxide (41 mg, 1.7 mmol) in i-propanol /THF (1:1, 4 mL) and water (1 mL) was stirred at 30oC for 1 h. The mixture was evaporated in vacuo and the e was diluted with water (5 mL). It was then extracted with ethyl acetate (3 X 10 mL). The combined ethyl acetate extract was concentrated under 391 reduced re and the residue was purified by reverse-phase prep-HPLC to afford 272 (52 mg, 50%) as a white solid. MS-ESI: [M+H]+ 622.3. 1H NMR (500 MHz, CDCl3) δ 8.77 (d, J = 2.0 Hz, 1H), 8.52 (d, J = 5.0 Hz, 1H), 8.19 (d, J = 1.5 Hz, 1H), 7.94 (s, 1H), 7.90 (d, J = 2.0 Hz, 1H), 7.61 (dd, J =2.0, 8.5 Hz,1H), 7.38 (d, J = 5.0 Hz, 1H), 6.86-6.84 (m, 2H), 5.12-5.09 5 (m, 1H), 4.77-4.74 (m, 2H), 4.69-4.66 (m, 1H), 4.61-4.59 (m, 2H), 4.54 (bs, 1H), 4.36-4.32 (m, 1H), 4.19-4.17 (m, 2H), 3.90-3.83 (m, 2H), 3.80-3.77 (m, 2H), 3.74 (s, 3H), 3.71-3.68 (m, 1H), 3.30-3.27 (m,2H), 2.60-2.59 (m, 2H), 2.54 (s, 2H), 1.30 (s, 6H).
Example 273a [4-(5-Bromomethyloxo-1,6-dihydropyridinyl){4,4- dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl]methyl 10 acetate 273a Br AcO O N N N O N 273a A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with 3-bromoiodomethylpyridin-2(1H)-one 214b (1.57 g, 5.0 mmol), {3- [(acetyloxy)methyl]{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dien- 15 10-yl}pyridinyl}boronic acid 199e (1.98 g, 5.0 mmol), PdCl2(dppf) (205 mg, 0.25 mmol), K3PO4 (2.12 g, 10.0 mmol), sodium acetate (820 mg, 10.0 mmol), acetonitrile (45 mL), and water (1 mL). The system was evacuated and refilled with N2. The reaction mixture was stirred at 30oC for 3 h. It was then filtered and the filtrate was concentrated under reduced re. The resulting residue was purified by -gel column chromatography eluting 20 with 30:1 romethane/methanol to afford 273a (580 mg, 22%) as a white solid. MS-ESI: [M+H]+ 539.2. 1H NMR (500 MHz, CDCl3) δ 8.49 (d, J = 5.0 Hz, 1H), 7.84 (d, J = 2.5 Hz, 1H), 7.45 (d, J = 2.0 Hz, 1H), 7.09 (d, J = 5.0 Hz, 1H), 6.79 (s, 1H), 5.15 (s, 2H), 4.55-4.51 (m, 1H), .25 (m, 1H), 4.15-4.13 (m, 1H), .04 (m, 1H), 3.68 (s, 3H), 2.58-2.56 (m, 2H), 2.51 (s, 2H), 1.86 (s, 3H), 1.28 (s, 6H). 25 Example 273b (2'-(7,7-dimethyloxo-3,4,7,8-tetrahydro-1H- cyclopenta[4,5]pyrrolo[1,2-a]pyrazin-2(6H)-yl)methyloxo(pyridinylamino)-1,6- dihydro-[3,4'-bipyridin]-3'-yl)methyl acetate 273b 392 Into a 1-dram vial was added 273a (40 mg, 0.074 mmol), 2-aminopyridine, (1.2 , cesium carbonate (1.5 equiv), os (10 mol%) and tris(dibenzylideneacetone) adium(0) (5 mol%) in dry 1,4-dioxane (0.2 M). The reaction was then stirred at 80°C 5 for 3 hours. After cooling to room temperature, the reaction was then diluted with dichloromethane (3 mL) and washed with water (2 x 3 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The crude t 273b was then carried on to the subsequent step without purification.
Example 273 3-[3-(hydroxymethyl)[1-methyloxo(2-pyridylamino) 10 pyridyl]pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazin one 273 Into a 1 dram vial was added 273b (1 equiv) in a 4:1 mixture of THF and water (1 mL). Lithium hydroxide (1.5 equiv) was then added to the mixture and the reaction was stirred at room temperature for 16 hours. The reaction was then diluted with dichloromethane 15 (3 mL) and washed with water (2 x 3 mL). The organic layer was collected, dried over ium sulfate, filtered, and concentrated in vacuo. The crude material was purified by reverse-phase chromatography to give 273. 1H NMR (400 MHz, DMSO-d6) δ 8.73 (d, J = 2.4 Hz, 1H), 8.62 (s, 1H), 8.49 (d, J = 5.0 Hz, 1H), 8.17 (dd, J = 5.1, 1.9 Hz, 1H), 7.62 – 7.56 (m, 1H), 7.54 (d, J = 2.5 Hz, 1H), 7.35 (d, J = 5.1 Hz, 1H), 7.30 (d, J = 8.5 Hz, 1H), 6.80 (dd, 20 J = 7.0, 5.1 Hz, 1H), 6.56 (s, 1H), 4.96 – 4.93 (m, 1H), 4.48 – 4.39 (m, 2H), 4.24 – 4.17 (m, 2H), 3.89 – 3.84 (m, 1H), 3.61 (s, 3H), 2.58 (d, J = 8.0 Hz, 2H), 2.43 (s, 3H), 1.22 (s, 6H). e 274 3-[3-(hydroxymethyl)[1-methyl[(5-methylpyrazinyl)amino] oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- b]pyrazinone 274 25 Following the procedures of Example 273, and substituting 2-amino methylpyrazine for 2-aminopyridine, 274 was prepared. 1H NMR (400 MHz, DMSO-d6) δ 8.98 (s, 1H), 8.68 – 8.58 (m, 2H), 8.49 (d, J = 5.1 Hz, 1H), 8.04 (s, 1H), 7.58 (d, J = 2.4 Hz, 1H), 7.34 (d, J = 5.1 Hz, 1H), 6.56 (s, 1H), 5.75 (s, 1H), 4.93 (t, J = 5.2 Hz, 1H), 4.47 – 4.37 393 (m, 2H), 4.25 – 4.16 (m, 2H), 3.88 – 3.82 (m, 1H), 3.62 (s, 3H), 2.57 (d, J = 8.0 Hz, 2H), 2.43 (s, 2H), 2.34 (s, 3H), 1.22 (s, 6H).
Example 275 3-[4-[5-[(5-fluoropyridyl)amino]methyloxopyridyl] (hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- 5 b]pyrazinone 275 Following the procedures of Example 273, and tuting 2-aminofluoropyridine for 2-aminopyridine, 275 was prepared. 1H NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1H), 8.65 (d, J = 2.4 Hz, 1H), 8.49 (d, J = 5.1 Hz, 1H), 8.14 (d, J = 3.1 Hz, 1H), 7.58 (td, J = 8.7, 3.1 Hz, 1H), 7.54 (d, J = 2.4 Hz, 1H), 7.40 (dd, J = 9.2, 3.9 Hz, 1H), 7.35 (d, J = 5.0 Hz, 1H), 10 6.56 (s, 1H), 4.98 – 4.91 (m, 1H), 4.46 – 4.38 (m, 2H), 4.23 – 4.16 (m, 2H), 3.88 – 3.82 (m, 1H), 3.61 (s, 3H), 2.61 – 2.51 (m, 2H), 2.43 (s, 2H), 1.22 (s, 6H).
Example 276 6-[[5-[2-(7,7-dimethyloxo-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinyl)(hydroxymethyl)pyridyl] methyloxopyridyl]amino]pyridinecarbonitrile 276 15 Following the procedures of Example 273, and substituting 2-aminocyanopyridine for 2-aminopyridine, 276 was prepared. 1H NMR (400 MHz, DMSO-d6) δ 9.38 (s, 1H), 8.73 (d, J = 2.2 Hz, 1H), 8.58 (d, J = 2.4 Hz, 1H), 8.49 (d, J = 5.0 Hz, 1H), 7.94 (dd, J = 9.0, 2.4 Hz, 1H), 7.68 (d, J = 2.4 Hz, 1H), 7.44 (d, J = 8.9 Hz, 1H), 7.36 (d, J = 5.2 Hz, 1H), 6.56 (s, 1H), 5.75 (s, 1H), 4.97 (t, J = 5.3 Hz, 1H), 4.46 – 4.38 (m, 2H), 4.21 (s, 3H), 3.84 (s, 1H), 20 3.62 (s, 3H), 2.58 (d, J = 8.1 Hz, 2H), 2.45 (s, 2H), 1.22 (s, 6H). e 277 hydroxymethyl)[5-[(5-methoxypyridyl)amino]methyl oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- b]pyrazinone 277 Following the procedures of Example 273, and substituting 2-amino 25 methoxypyridine for 2-aminopyridine, 277 was prepared. 1H NMR (400 MHz, DMSO-d6) δ 8.61 (d, J = 2.4 Hz, 1H), 8.51 – 8.47 (m, 2H), 7.91 (d, J = 2.4 Hz, 1H), 7.48 (d, J = 2.4 Hz, 1H), 7.34 (d, J = 5.0 Hz, 1H), 7.33 – 7.29 (m, 2H), 6.56 (s, 1H), 4.94 (t, J = 5.3 Hz, 1H), 4.47 – 4.38 (m, 2H), 4.24 – 4.16 (m, 2H), 3.87 – 3.83 (m, 1H), 3.75 (s, 3H), 3.60 (s, 3H), 2.58 (d, J = 7.9 Hz, 2H), 2.43 (s, 2H), 1.22 (s, 6H). 30 Example 278 3-[4-[5-[(5-cyclopropylpyridyl)amino]methyloxopyridyl]- 3-(hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- zinone 278 Following the procedures of Example 273, and substituting 2-amino cyclopropylpyridine for 2-aminopyridine, 278 was prepared. 1H NMR (400 MHz, DMSO- 394 d6) δ 8.67 (d, J = 2.4 Hz, 1H), 8.53 – 8.45 (m, 2H), 8.01 (d, J = 2.5 Hz, 1H), 7.50 (d, J = 2.4 Hz, 1H), 7.34 (d, J = 5.0 Hz, 1H), 7.28 (dd, J = 8.6, 2.5 Hz, 1H), 7.21 (d, J = 8.6 Hz, 1H), 6.56 (s, 1H), 4.96 – 4.92 (m, 1H), 4.47 – 4.41 (m, 2H), 4.24 – 4.16 (m, 2H), 3.89 – 3.85 (m, 1H), 3.60 (s, 3H), 2.58 (d, J = 7.9 Hz, 2H), 2.44 (s, 2H), 1.86 – 1.77 (m, 1H), 1.22 (s, 6H), 5 0.93 (t, J = 8.0 Hz, 1H), 0.90 – 0.85 (m, 1H), 0.67 – 0.58 (m, 2H).
Example 279 3-[3-(hydroxymethyl)[1-methyloxo[[5-(trifluoromethyl) pyridyl]amino]pyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 279 ing the procedures of Example 273, and substituting 2-amino 10 trifluoromethylpyridine for 2-aminopyridine, 279 was prepared. 1H NMR (400 MHz, DMSO-d6) δ 9.23 (s, 1H), 8.80 (d, J = 2.4 Hz, 1H), 8.54 – 8.46 (m, 2H), 7.88 (dd, J = 8.7, 2.6 Hz, 1H), 7.66 (d, J = 2.3 Hz, 1H), 7.48 (d, J = 8.8 Hz, 1H), 7.37 (d, J = 5.1 Hz, 1H), 6.56 (s, 1H), 4.97 (t, J = 5.1 Hz, 1H), 4.50 – 4.40 (m, 2H), 4.26 – 4.16 (m, 2H), 3.90 – 3.81 (m, 1H), 3.62 (s, 3H), 2.58 (d, J = 8.2 Hz, 2H), 2.43 (s, 2H), 1.22 (s, 6H). 15 Example 280 hydroxymethyl)[1-methyl[[1-methyl(morpholine carbonyl)pyrazolyl]amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 280 Following the procedures of Example 273, and substituting (3-aminomethyl-1H- pyrazolyl)(morpholino)methanone for 2-aminopyridine, 280 was prepared. 1H NMR (400 20 MHz, DMSO-d6) δ 8.48 (d, J = 5.0 Hz, 1H), 8.27 (s, 1H), 8.05 (d, J = 2.4 Hz, 1H), 7.43 (d, J = 2.4 Hz, 1H), 7.33 (d, J = 5.1 Hz, 1H), 6.55 (s, 1H), 6.30 (s, 2H), 4.99 – 4.91 (m, 1H), 4.48 – 4.39 (m, 2H), 4.23 – 4.15 (m, 7H), 3.89 – 3.82 (m, 2H), 3.72 (s, 3H), 3.59 (s, 3H), 3.27 (s, 2H), 2.58 (d, J = 7.5 Hz, 2H), 2.43 (s, 2H), 1.22 (s, 6H).
Example 281 3-[3-(hydroxymethyl)[1-methyl[(5-methylpyridyl)amino] 25 pyridyl]pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- b]pyrazinone 281 Following the procedures of Example 273, and substituting 2-aminomethylpyridine for 2-aminopyridine, 281 was prepared. 1H NMR (400 MHz, DMSO-d6) δ 8.67 (d, J = 2.4 Hz, 1H), 8.48 (d, J = 5.4 Hz, 2H), 8.01 (d, J = 2.3 Hz, 1H), 7.51 (d, J = 2.4 Hz, 1H), 7.44 (dd, 30 J = 8.5, 2.4 Hz, 1H), 7.34 (d, J = 5.1 Hz, 1H), 7.21 (d, J = 8.5 Hz, 1H), 6.56 (s, 1H), 5.75 (s, 1H), 4.93 (t, J = 5.4 Hz, 1H), 4.46 – 4.36 (m, 2H), 4.26 – 4.16 (m, 2H), 3.86 – 3.80 (m, 1H), 3.60 (s, 3H), 2.58 (d, J = 7.8 Hz, 2H), 2.43 (s, 2H), 2.17 (s, 3H), 1.22 (s, 6H).
Example 282a 3,3-Dimethylcyclopentanone 282a 395 O HS THF, LiOH DMF, POCl3 O 65 °C, 5h O O O Cl 1. Et3N, DCM, reflux, 16h S 2. EtOH, Et3N, reflux, 12h O 282a 282b 282c 1. n-BuLi, THF, CHO 1. SOCl2, reflux -78 °C, 6 h H OH N H S 2. t-BuNH2, DCM S t-Bu 2. DMF, -78 °C, 16 h S N t- O rt, 16 h O Bu O 282d 282e 282f O Br Cl 30% of H2SO4 NH2NH2H2O (3.0 eq.) NH2 reflux, 16 h N N RT, 3 h N H S NH S N CuBr, Sarcosine, K2CO3 t-Bu O 95 °C, 15 h O 282h 282g O N N O 191j N N NH S N Cl Pd(dppf)Cl2, K3PO4, THF O O H2O reflux, 6h N O N S N N 282i O N 282j To a sion of CuI (81.0 g, 420 mmol) in anhydrous ethyl ether (500 mL) cooled to 0 oC was added the solution of methyllithium in ethyl ether (430 mL, 860 mmol, 2.0M) over a period of 30 minutes. The mixture was stirred at 0oC for 2 h. To the above mixture was 5 added 3-methylcyclopentenone (33.6 g, 350 mmol) dropwise over a period of 1 h at 0 oC.
The ing mixture was d at 0oC for another 2 h. It was then quenched with saturated NH4Cl (300 mL) and filtered. The filtrate was extracted with ethyl ether (2 X 200 mL). The combined organic layer was dried over anhydrous Mg2SO4 and filtered. The filtrate was evaporated under reduced pressure to afford 282a as a colorless oil (28 g, 71%). 1H NMR 396 (500 MHz, DMSO-d6) δ 2.31 (t, J = 8.0 Hz, 2H), 2.05 (s, 2H) , 1.79 (t, J = 8.0 Hz, 2H), 1.12 (s, 6H).
Example 282b 2-Chloro-4,4-dimethylcyclopentenecarbaldehyde 282b To a solution of DMF (18.3 g, 250 mmol) in dichloromethane (300 mL) cooled to 0oC 5 was added POCl3 (40.5 g, 250 mmol) over a period of 10 minutes. The mixture was stirred at 20oC for 1 h. To the above mixture was added 282a (28.0 g, 250 mmol) dropwise over a period of 20 minutes. The resulting mixture was heated at reflux for 20 h. The reaction mixture was cooled to room ature and poured into a solution of sodium e (60 g) in ice-water (400 g). The mixture was extracted with dichloromethane (2 X 300 mL). The 10 combined organic layer was washed with water (2 X 200 mL), dried over ous Mg2SO4 and filtered. The filtrate was evaporated under reduced re to afford 282b as a colorless oil (33.0 g, crude). 1H NMR (500 MHz, DMSO-d 6) δ 9.99 (s, 1H), 2.62 (d, J = 2.0 Hz, 2H), 2.38 (d, J = 2.0 Hz, 2H), 1.15 (s, 6H).
Example 282c Ethyl methyl-5,6-dihydro-4H-cyclopenta[b]thiophene 15 carboxylate 282c To a solution of 282b (33.0 g, crude) in dichloromethane (400 mL) and triethylamine (60 g, 600 mmol) was added ethyl 2-mercaptoacetate (19.2 g, 160 mmol). The reaction e was heated at reflux for 6 h. It was then concentrated under reduced pressure. The e was dissolved in ethanol (400 mL) and triethylamine (60 g, 600 mmol). The mixture 20 was heated at reflux for 12 h. It was concentrated again under reduced pressure and the residue was purified by silica-gel column chromatography eluting with 40:1 petroleum ether/ethyl acetate to afford 282c as yellow solid (18.0 g, 32%, over two steps). MS-ESI: [M+H]+ 225.3. 1H NMR (500 MHz, 6) δ 7.49 (s, 1H), 4.32 (q, J = 7.0 Hz, 2H), 2.72 (s, 2H), 2.56 (s, 2H), 1.35 (t, J = 7.0 Hz, 3H),1.22 (s, 6H). 25 Example 282d 5,5-Dimethyl-5,6-dihydro-4H-cyclopenta[b]thiophene carboxylic acid 282d To the solution of 282c (16.0 g, 71.0 mmol) in propanol (200 mL), tetrahydrofuran (200 mL), and water (200 mL) was added lithium hydroxide (6.82 g, 284 mmol). The reaction mixture was heated at 65oC for 5 h. The organic solvents were removed under 30 reduced pressure. The pH of the residue was adjusted to 1.0 with hydrochloride acid (12M).
The precipitate was collected by filtration and dried in vacuo to afford 282d (12.0 g, 86%) as white solid. MS-ESI: [M+H]+ 196.9 Example 282e N-tert-Butyl-5,5-dimethyl-5,6-dihydro-4H- cyclopenta[b]thiophenecarboxamide 282e 397 A suspension of 282d (12.0 g, 61.0 mmol) in SOCl2 (80 mL) was heated at 65oC for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was diluted with dichloromethane (20 mL), which was added to the solution of ylpropanamine (4.45 g, 61.0 mmol) and ylamine (18.0 g, 180 mmol) in dichloromethane (180 mL). The 5 resulting mixture was stirred for 16 h and diluted with dichloromethane (200 mL). It was washed with water (3 X 50 mL), dried over anhydrous Mg2SO4, filtered, and evaporated under reduced pressure to afford 282e (15.0 g, 97%) as yellow solid. MS-ESI: [M+H]+ 252.0 Example 282f -Butylformyl-5,5-dimethyl-5,6-dihydro-4H- cyclopenta[b]thiophenecarboxamide 282f 10 To a on of 282e (1.5 g, 6.0 mmol) in anhydrous THF (60 mL) cooled at -70oC was added the solution of n-butyl m (10.0 mL, 25 mmol, 2.5 M in hexane) over a period of 5 minutes. It was stirred at -70 oC for 6 h. DMF (1.3 g, 18.0 mmol) was added over a period of 5 minutes and the result mixture was stirred at room temperature for overnight. It was then quenched with saturated NH4Cl (40 mL) and concentrated under reduced pressure. 15 The residue was extracted with ethyl acetate (2 X 30 mL). The combined organic layer was dried over anhydrous Mg2SO4 and filtered. The filtrate was evaporated under reduced pressure to afford 282f as yellow solid (1.34 g, 80%). MS-ESI: [M+H]+ 280.3 Example 282g N-tert-Butyl(hydrazonomethyl)-5,5-dimethyl-5,6-dihydro- 4H-cyclopenta[b]thiophenecarboxamide 282g 20 To a solution of 85% aqueous hydrazine (10 mL) in THF (180 mL) was added 282f (5.6 g, 20.0 mmol) in anhydrous THF (20 mL) over a period of 5 minutes. It was stirred at 20oC for 3 h. The reaction mixture was concentrated under reduced pressure to afford 282g as black solid (6.0 g, yield: 95%, purity: 95%). MS-ESI: [M+H]+ 294.0 Example 282h 4,4-Dimethylthia-10,11-diazatricyclo[6.4.0.02,6]dodeca- 25 1(8),2(6),11-trienone 282h A solution of 282g (3.8 g, 13.0 mmol) in 30% H2SO4 (100 mL) was heated at reflux for 16 h. The reaction mixture was cooled to room temperature and ted with dichloromethane (3 X 200 mL). The combined organic layer was concentrated under reduced re and the residue was purified by silica-gel column chromatography eluting with 30 100:1 dichloromethane/methanol to afford 282h as yellow solid (1.72 g, 60%). MS-ESI: [M+H]+ 221.0 Example 282i 4-Chloro{4,4-dimethyloxothia-10,11- diazatricyclo[6.4.0.02,6]dodeca-1(8),2(6),11-trienyl}pyridinecarbaldehyde 282i 398 Following the procedures as described in Example 108a, and starting with 282h (330 mg, 1.5 mmol) and 2-bromochloronicotinaldehyde (950 mg, 4.5 mmol), 282i was obtained as a yellow solid (260 mg, 48%). MS-ESI: [M+H]+ 359.9 Example 282j 2-{4,4-Dimethyloxothia-10,11- 5 diazatricyclo[6.4.0.02,6]dodeca-1(8),2(6),11-trienyl}[1-methyl({5-[(2S)methyl (oxetanyl)piperazinyl]pyridinyl}amino)oxo-1,6-dihydropyridinyl]pyridine carbaldehyde 282j Following the procedure for preparation in Example 191k, and starting with 282i (216 mg, 0.60 mmol), and (S)methyl(5-(2-methyl(oxetanyl)piperazinyl)pyridin 10 o)(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 191j (482 mg, 0.90 mmol), 282j was obtained as a yellow solid (407 mg, 48%). MS-ESI: [M+H]+ 678.8 Example 282 3-[3-(hydroxymethyl)[1-methyl[[5-[(2S)methyl(oxetan yl)piperazinyl]pyridyl]amino]oxopyridyl]pyridyl]-7,7-dimethyl-6,8- dihydrocyclopenta[3,4]thieno[1,3-d]pyridazinone 282 15 Following the procedures in Example 191, and ng with 282j (370 mg, 0.55 mmol), 282 was ed as a yellow solid (64 mg, 17%). MS-ESI: [M+H]+ 681.3. 1H NMR (500 MHz, DMSO-d6) δ 8.63 (d, J = 2.0 Hz, 1H), 8.56 (d, J = 5.0 Hz, 1H), 8.46-8.45 (m, 2H), 7.85(d, J = 3.0 Hz, 1H), 7.53 (d, J = 5.0 Hz,1H), 7.47 (d, J = 2.5 Hz, 1H), 7.36 (dd, J = 3.0, 9.0 Hz, 1H), 7.24 (d, J = 9.0 Hz, 1H), 4.85 (t, J = 5.0 Hz, 1H), 4.57-4.54 (m, 2H), 4.47 (t, J = 20 6.0 Hz, 1H), 4.4-4.37 (m, 3H), 3.68-3.67 (m, 1H), 3.60 (s, 3H), 3.40-3.38 (m, 1H), 3.11-3.08 (m, 1H), 2.96-2.90 (m, 3H), 2.81-2.79 (m, 2H), 2.56-2.53 (m, 1H), 2.33-2.32 (m, 2H), 2.19- 2.16 (m, 1H), 1.28 (s, 3H), 1.27 (s, 3H), 0.93 (d, J = 6.0 Hz, 3H).
Example 283a 5-Bromomethyl(5-methylisoxazolylamino)pyridin- 2(1H)-one 283a 25 A 100-mL single-neck round-bottomed flask ed with a magnetic stirrer and a reflux condenser was charged with 5-methylisoxazolamine (1.0 g, 10.2 mmol), 3,5- dibromomethylpyridin-2(1H)-one (4.09 g, 15.3 mmol), Pd2(dba)3 (467 mg, 0.51 mmol), 399 Xantphos (598 mg, 1.02 mmol), Cs2CO3 (6.65 g, 20.4 mmol), and dioxane (50 mL). After three cycles of vacuum/argon flush, the reaction mixture was heated at 100oC for 3 h.
Analysis of the reaction mixture by LCMS showed complete conversion to the d product. It was filtered when the mixture was still hot. The filtrate was cooled down to room 5 temperature and the ing precipitation was collected by filtration to afford 283a (1.6 g, 55%) as a yellow solid. MS-ESI: [M+H]+ 284.1 Example 283b (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- -dien yl}{1-methyl[(5-methyl-1,2-oxazolyl)amino]oxo-1,6- dihydropyridinyl}pyridinyl)methyl Acetate 283b 10 A 50-mL round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 283a (150 mg, 0.53 mmol), {3-[(acetyloxy)methyl]{4,4- dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (834 mg, 2.1 mmol), Pd(dppf)Cl2 (21 mg, 0.026 mmol), K3PO4 (224 mg, 0.053 mmol), sodium acetate (87 mg, 1.1 mmol), acetonitrile (10 mL), and water (5 drops). After 15 three cycles of vacuum/N2 flush, the e was heated at 100oC for 1 h. Then it was cooled to room temperature and ed. The filtrate was concentrated under reduced pressure and the residue was washed with acetonitrile to afford the 283b (100 mg, 34%) as white solid.
MS-ESI: [M+H]+ 557.3 Example 283 3-[3-(hydroxymethyl)[1-methyl[(5-methylisoxazolyl)amino]- 20 6-oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- b]pyrazinone 283 To a solution of 283b (90 mg, 0.162 mmol) in THF (5 mL), i-propanol (5 mL), and water (5 mL) was added lithium hydroxide (3.8 mg, 1.62 mmol). The reaction mixture was stirred at room temperature for 1 h and concentrated under reduced pressure. The residue was 25 extracted with dichloromethane (20 mL×3). The combined c layer was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase PLC to afford 283 (65 mg, 78%) as white solid. MS-ESI: [M+H]+ 514.9. 1H NMR (500 MHz, DMSO-d6) δ 8.98 (s, 1H), 8.49 (d, J = 5.0 Hz, 1H), 8.02 (d, J = 2.0 Hz, 1H), 7.56 (d, J = 2.5 Hz, 1H), 7.32 (d, J = 5.0 Hz, 1H), 6.57 (s, 1H), 6.25 (s, 30 1H), 4.93 (t, J = 5.0 Hz, 1H), .38 (m, 2H), 4.25-4.19 (m, 3H), 3.87-3.85 (m, 1H), 3.60 (s, 3H), .54 (m, 2H), 2.43 (s, 2H), 2.31 (s, 3H), 1.22 (s, 6H).
Example 284a 2-(10-Fluorooxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol- 2(1H)-yl) (1-methyl(5-(oxetanyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin ylamino)oxo-1,6-dihydropyridinyl)nicotinaldehyde 284a 400 O N N N NH O O N N N F O N 284a A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and reflux condenser was charged with1-methyl(5-(oxetanyl)-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)(4,4,5,5-tetramethyl-1,3,2-dioxaborolan 5 yl)pyridin-2(1H)-one 163a (354 mg, 0.83 mmol), 4-chloro(10-fluorooxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)nicotinaldehyde 134c (289 mg, 0.83 mmol), PdCl2(dppf) (68 mg, 0.08 mmol), K3PO4 (352 mg, 1.66 mmol), sodium acetate (136 mg, 1.66 mmol), itrile (50 mL), and water (3 mL). The system was evacuated and refilled with N2. The reaction e was heated at 100ºC for 2 h. It was then cooled to room temperature 10 and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 284a (305 mg, 60%) as a brown solid. MS-ESI: [M+H]+:613.6.
Example 284 10-fluoro[3-(hydroxymethyl)[1-methyl[[5-(oxetanyl)-6,7- 15 dihydro-4H-pyrazolo[1,5-a]pyrazinyl]amino]oxopyridyl]pyridyl]-3,4,6,7,8,9- hexahydropyrido[3,4-b]indolizinone 284 To a suspension of 284a (250 mg, 0.41 mmol) in methanol (20 mL) was added sodium borohydride (47 mg, 1.23 mmol) at 0oC. The mixture was d for 30 minutes. It was then quenched with water (2 mL) and concentrated under reduced pressure. The residue 20 was purified by reverse-phase PLC to afford 284 (20 mg, 6.6 %). MS-ESI: [M+H]+ 615.6. 1H NMR (500 MHz, CDCl3) δ 8.46 (d, J = 5.0 Hz, 1H), 7.94 (d, J = 3.0 Hz, 1H), 7.66 (d, J = 2.0 Hz, 1H), 7.43 (s, 1H), 7.34 (d, J = 5.0 Hz, 1H), 5.75 (s, 1H), 4.95 (t, J = 6.5 Hz, 1H), 4.76-4.74 (m, 2H), .65-4.67 (m, 3H), 4.46-4.44 (m, 1H), 4.35-4.33 (m, 1H), 4.10- 4.08 (m, 4H), 3.38-3.35 (m, 2H), 3.69 (s, 3H), 3.58-3.56 (m, 2H), 2.842.82 (m, 2H), 2.58- 25 2.53 (m, 4H), 1.89-1.84 (m, 2H), 1.77-1.76 (m, 2H).
Example 285a 4-Nitro(oxetanyl)-1H-imidazole 285a 401 O N N NH O O AcO O N H2,Pd/ C, 273a N N N N MeOH,rt, 1 h Pd2(dba)3 (0.05 eq), xantphos (0.1 eq), O N N NO2 N NH2 Cs2CO3 (2 eq), e, 100oC, 2h 285a 285b 285c A sealed tube was charged with o-1H-imidazole (500 mg, 4.42 mmol), 3- iodooxetane (920 mg, 5.0 mmol), Cs2CO3 (2.90 g, 8.84 mmol), and dioxane (12 mL). The sealed tube was heated at 120oC for 16 h. It was then cooled to room temperature and filtered. 5 The filtrate was evaporated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 285a as a white solid (250 mg, 33%). MS-ESI: [M+H]+ 170.2.
Example 285b 1-(Oxetanyl)-1H-imidazolamine 285b A 25-mL single-neck round-bottomed flask was purged with nitrogen and charged 10 with 285a (100 mg, 0.6 mmol), 10% palladium on carbon (10% wet, 10 mg) and methanol (10 mL). The flask was evacuated, charged with hydrogen gas (via balloon), and stirred for 1 h at room temperature. The hydrogen was then evacuated and nitrogen was charged to the flask. The catalyst was removed by filtration through a pad of CELITE® and the te was concentrated under reduced pressure to afford 285b (70 mg, 85%). : [M+H]+ 140.3. 15 Example 285c (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}(1-methyl{[1-(oxetanyl)-1H-imidazolyl]amino}oxo-1,6- dihydropyridinyl)pyridinyl)methyl Acetate 285c A 25-mL round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was d with 285b (40 mg, 0.28 mmol), [4-(5-bromomethyloxo-1,6- 20 opyridinyl){4,4-dimethyloxo-1,10-diazatri-cyclo[6.4.0.02,6]dodeca-2(6),7- dienyl}pyridinyl]methyl acetate 273a (150 mg, 0.28 mmol), Pd2(dba)3 (15 mg, 0.015 mmol), XantPhos (18 mg, 0.03 mmol), cesium carbonate (200 mg, 0.6 mmol), and 1,4- dioxane (6 mL). After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 2 h. It was then cooled to room temperature and filtered. The filtrate was trated 25 under reduced pressure and the residue was purified by -gel column chromatography eluting with 25:1 dichloromethane/methanol to afford 285c as a yellow solid (80 mg, 47%).
MS-ESI: [M+H]+ 598.3. 402 Example 285 3-[3-(hydroxymethyl)[1-methyl[[1-(oxetanyl)imidazol no]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 285 A mixture of 285c (80 mg, 0.13 mmol) and lithium hydroxide·water (55 mg, 1.3 5 mmol) in anol /THF (3:2, 5 mL) and water (2 mL) was stirred at 30oC for 1 h. The mixture was evaporated under reduced pressure and water (5 mL) was added to the residue. It was then extracted with dichloromethane (3 X 10 mL). The combined dichloromethane extract was concentrated under reduced pressure and the residue was purified by reversephase prep-HPLC to afford 285 (36 mg, 50%) as a white solid. MS-ESI: [M+H]+ 556.3. 1H 10 NMR (500 MHz, CDCl3) δ 8.48 (d, J = 5.0 Hz, 1H), 7.53 (d, J = 2.0 Hz, 1H), 7.47-7.45 (m, 2H), 7.35-7.34 (m, 2H), 7.28 (s, 1H), 6.88 (s, 1H), 5.27-5.19 (m, 2H), 5.10-5.07 (m, 2H), 4.94-4.91 (m, 2H), 4.69-4.65 (m, 1H), 4.52-4.44 (m, 2H), 4.17-4.16 (m, 2H), 3.87-3.84 (m, 1H), 3.72 (s, 3H), 2.59-2.58 (m, 2H), 2.53 (s, 2H), 1.29 (s, 6H).
Example 286a (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 15 2(6),7-dienyl}{1-methyl[(1,2-oxazolyl)amino]oxo-1,6-dihydropyridin yl}pyridinyl)methyl Acetate 286a A 25-mL round-bottomed flask equipped with a reflux condenser was charged with bromomethyloxo-1,6-dihydropyridinyl){4,4-dimethyloxo-1,10- 20 diazatricy-clo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl]methyl e 273a (161 g, 0.30 mmol), isoxazolamine (25 mg, 0.30 mmol), cesium carbonate (196 mg, 0.60 mmol), and 1,4-dioxane (10 mL). After bubbling en through the suspension for 10 minutes, tris(dibenzylideneacetone)dipalladium(0) (14.0 mg, 0.015 mmol) and xantphos (17 mg, 0.030 mmol) were added. The system was subjected to three cycles of vacuum/argon flush and 25 heated at reflux for 5 h. It was then cooled to room temperature and ed. The solid was washed with dichloromethane (2 X 10 mL). The combined organic filtrate was concentrated under reduced pressure. The e was purified by silica-gel column chromatography 403 eluting with dichloromethane/methanol (80/1 to 30/1) to afford 286a (96 mg, 59%) as yellow solid. MS-ESI: [M+H]+ 542.8.
Example 286 3-[3-(hydroxymethyl)[5-(isoxazolylamino)methyloxo pyridyl]pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazin 5 one 286 To a solution of 286a (96 mg, 0.18 mmol) in THF/i-propanol /water(5/3/2mL) was added lithium hydroxide (21 mg, 0.88 mmol). The mixture was stirred at room temperature for 1 h. After the reaction was te, the mixture was evaporated under pressure and the residue was ed by e-phase prep-HPLC to afford 286 as a white solid (75 mg, 10 85%). MS-ESI: [M+H]+ 501.3. 1H NMR (500 MHz, CDCl3) δ 8.49 (d, J = 6.5 Hz, 1H), 8.19 (d, J = 2.5 Hz, 1H), 8.10 (d, J = 2.5 Hz, 1H), 8.97 (d, J = 3.0 Hz, 1H), 7.71 (s, 1H), 7.37 (d, J = 6.5 Hz, 1H), 6.85 (s, 1H), 6.18 (d, J = 2.5 Hz, 1H), 5.12-5.11 (m, 1H), 4.66-4.64 (m, 1H), 4.52-4.51 (m, 1H), 4.29-4.27 (m, 1H), 4.18-4.16 (m, 2H), 3.87-3.86 (m, 1H), 3.73 (s, 3H), 2.59-2.57 (m, 2H), 2.53-2.51 (m, 2H), 1.28 (s, 6H). 15 Example 287a N-Methoxy-N-methylnitro-1H-pyrazolecarboxamide 287a 404 O O O O N N O N O O NHBoc H2, Pd/C O Cl HN HN MeOH HN toluene, 130°C N N Na2CO3 N NO2 N NH2 p-TsOH 287a 287b 287c O N O O N BocHN BocHN N N N MeMgBr N CF3COOH N N NaBH4 N N N THF CH2Cl2 MeOH 287e 287f 287d N HN N N N N NH2OH.HCl N 10:1 MeOH/AcOH N EtOH NaBH3CN, HCHO 287h 287g N N N N NH AcO O N N 273a N NH2 N N Pd2(dba)3, Xantphos O N 287i Cs2CO3, dioxane 287j A 500-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with 3-nitro-1H-pyrazolecarboxylic acid (15.7 g, 1.0 eq., 100 mmol), N,O- dimethylhydroxylamine hydrochloride (19.5 g, 2.0 eq., 200 mmol), HATU (76.0 g, 2.0 eq., 5 200 mmol), triethylamine (40.4 g, 4.0 eq., 400 mmol), and dichloromethane (300 mL). The reaction mixture was stirred at room temperature for overnight. The solvent was removed under reduced re. The resulting residue was purified by silica-gel column tography eluting with 100:1 romethane/methanol to afford 287a (16.0 g, 80%) as white solid. MS-ESI: [M+H]+ 201.1 10 Example 287b 3-Amino-N-methoxy-N-methyl-1H-pyrazolecarboxamide 287b 405 A 250-mL single-neck round-bottomed flask was purged with nitrogen and charged with 287a (16.0 g, 1.0 eq., 80.0 mmol), 10% palladium on carbon (50% wet, 800 mg), and methanol (100 mL). The mixture was evacuated, charged with hydrogen gas, and stirred under hydrogen atmosphere at room temperature overnight. The en was then 5 evacuated and en was charged into the flask. The catalyst was removed by filtration through a pad of CELITE®. The filtrate was concentrated under reduced pressure to afford 287b (11.0 g, 81%) as white solid. MS-ESI: [M+H]+ 171.1 Example 287c 3-(2,5-Dimethyl-1H-pyrrolyl)-N-methoxy-N-methyl-1H- pyrazolecarboxamide 287c 10 A 250-mL round-bottomed flask equipped with a magnetic stirrer and a tark trap was charged with 287b (11.0 g, 1.0 eq., 64.7 mmol), hexane-2,5-dione (11.1 g, 1.5 eq., 97.2 mmol), p-toluenesulfonic acid monohydrate (558 mg, 0.05 eq., 3.24 mmol), and toluene (100 mL). The reaction mixture was refluxed overnight. The resulting mixture was cooled to room temperature and concentrated under reduced pressure. The resulting residue was 15 purified by silica-gel column chromatography eluting with 1:2 petroleum ether/ethyl acetate to afford 287c (10.4 g, 65%) as white solid. MS-ESI: [M+H]+ 249.0 Example 287d utyl 2-(3-(2,5-Dimethyl-1H-pyrrolyl) (methoxy(methyl)carbamoyl)-1H-pyrazolyl)ethylcarbamate 287d A 250-mL round-bottomed flask equipped with a magnetic stirrer was charged with 20 287c (10.4 g, 1.0 eq., 41.9 mmol), tert-butyl 2-chloroethylcarbamate (37.7 g, 5.0 eq., 210.0 mmol), Na2CO3 (22.3 g, 5.0 eq., 210.0 mmol), and DMF (100 mL). The on mixture was stirred at 110oC overnight. After cooling to room temperature, the resulting mixture was poured into water (200 mL) and extracted with ethyl acetate (3 X 100 mL). The combined organic layer was concentrated under reduced pressure. The resulting residue was ed by 25 silica-gel column chromatography eluting with ethyl e to afford 287d (10.8 g, 66%) as yellow oil. MS-ESI: [M+H]+ 392.0 Example 287e tert-Butyl 2-(5-Acetyl(2,5-dimethyl-1H-pyrrolyl)-1H- lyl)ethylcarbamate 287e A 250-mL round-bottomed flask equipped with a magnetic r was charged with 30 287d (7.82 g, 1.0 eq., 20.0 mmol) and THF (100 mL) under N2 protection. A solution of MeMgBr (3.0 M in ether) (17 mL, 2.5 eq., 50.0 mmol) was added at -78oC. The mixture was stirred at room temperature for 3 h and quenched with saturated NH4Cl solution. It was then concentrated under reduced pressure and the residue was ted with ethyl acetate (3 x 50 mL). The combined organic layer was evaporated under d pressure. The residue was 406 purified by silica-gel column chromatography eluting with 4:1 petroleum ether/ethyl acetate to afford 287e as ess oil (5.40 g, 78%). MS-ESI: [M+H]+ 347.0. 1H NMR (500 MHz, CDCl3) δ 6.57 (s, 1H), 5.91 (s, 2H), 4.93 (bs, 1H), 4.71 (t, J = 5.5 Hz, 2H), 3.62 (t, J = 5.5 Hz, 2H), 2.57 (s, 3H), 2.16 (s, 6H), 1.28 (s, 9H). 5 Example 287f 2-(2,5-Dimethyl-1H-pyrrolyl)methyl-6,7- dihydropyrazolo[1,5-a]pyrazine 287f A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with 287e(5.40 g, 1.0 eq., 15.6 mmol), CF3COOH (10 mL), and dichloromethane (50 mL). The mixture was stirred at room temperature for 1 h and concentrated under reduced 10 pressure to afford crude 287f, which was used in the next step without further purification.
MS-ESI: [M+H]+ 229.1 Example 287g 2-(2,5-Dimethyl-1H-pyrrolyl)methyl-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazine 287g A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer was 15 charged with 287f (3.56 g, 1.0 eq., 15.6 mmol), NaBH4 (2.96 g, 5.0 eq., 78.0 mmol), and ol (50 mL). The mixture was d at room ature for 1 h and concentrated under reduced pressure. The residue was partitioned between water (50 mL) and dichloromethane (50 mL). The water phase was extracted with dichloromethane (3 X 50 mL).
The combined organic layer was concentrated under reduced pressure. The residue was 20 purified by silica-gel column chromatography eluting with 10:1 dichloromethane/methanol to afford 287g as a colorless oil (1.54 g, 43% over two . : [M+H]+ 231.3. 1H NMR (500 MHz, CDCl3) δ 5.91 (s, 1H), 5.86 (s, 2H), 4.17-4.11 (m, 3H), 3.51-3.48 (m, 1H), 3.36- 3.31 (m, 1H), 2.13 (s, 6H), 1.50 (d, J = 6.5 Hz, 3H).
Example 287h 2-(2,5-Dimethyl-1H-pyrrolyl)-4,5-dimethyl-4,5,6,7- 25 tetrahydropyrazolo[1,5-a]pyrazine 287h A 100-mL -neck round-bottomed flask equipped with a magnetic stirrer was charged with 287g (1.54 g, 1.0 eq., 6.70 mmol), dehyde (37% in water) (1.09 g, 2.0 eq., 13.4 mmol), NaBH3CN (2.11 g, 5.0 eq., 33.5 mmol), HOAc (3 mL), and methanol (30 mL).
The mixture was stirred at room temperature for 1 h and concentrated under reduced pressure. 30 The residue was partitioned between water (50 mL) and dichloromethane (50 mL). The water phase was extracted with dichloromethane (3 X 50 mL). The combined c layer was concentrated under reduced pressure to afford crude 287h, which was used in the next step without further purification. MS-ESI: [M+H]+ 245.0 407 Example 287i 4,5-Dimethyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin amine 287i A 100-mL single-neck round-bottomed flask equipped with a reflux condenser was charged with 287h (1.63 g, 1.0 eq., 6.70 mmol), NH2OH.HCl (2.33 g, 5.0 eq., 33.5 mmol), 5 and ethanol (50 mL). The mixture was refluxed for 2 days. It was then cooled to room temperature and concentrated under reduced pressure. The e was purified by reversephase prep-HPLC to afford 287i as a yellow solid (211 mg, 19%). MS-ESI: [M+H]+ 167.1. 1H NMR (500 MHz, CDCl 3) δ 5.36 (s, 1H), 4.04-4.00 (m, 1H), 3.94-3.92 (m, 1H), 3.61 (bs, 2H), 3.30 (q, J = 6.5 Hz, 1H), 3.10-3.08 (m, 1H), 2.81-2.75 (m, 1H), 2.43 (s, 3H), 1.38 (d, J = 10 6.5 Hz, 3H). e 287j (4-(5-(4,5-Dimethyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin- 2-ylamino)methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl e 287j A 25-mL round-bottomed flask equipped with a magnetic stirrer and a reflux 15 condenser was charged with 287i (20 mg, 1.0 eq., 0.12 mmol), (4-(5-bromomethyloxo- 1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- yl)pyridinyl)methyl acetate 273a (127 mg, 2.0 eq., 0.24 mmol), Pd2(dba)3 (9.0 mg, 0.1 eq., 0.010 mmol), Xantphos (11 mg, 0.2 eq., 0.020 mmol), Cs2CO3 (78 mg, 2.0 eq., 0.24 mmol), and dioxane (5 mL). After three cycles of vacuum/N2 flush, the mixture was stirred at 100oC 20 for 2 h. It was then cooled to room temperature and filtered. The te was concentrated under d pressure. The resulting residue was purified by silica-gel column chromatography g with 20:1 dichloromethane/methanol to afford 287j as a brown solid (60 mg, 82%). MS-ESI: [M+H]+ 610.9 e 287 2-[4-[5-[(4,5-dimethyl-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazin 25 yl)amino]methyloxopyridyl](hydroxymethyl)pyridyl]-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indolone 287 A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with 287j (60 mg, 1.0 eq., 0.098 mmol), lithium hydroxide (12 mg, 5.0 eq., 0.49 mmol), i-propanol/THF (4/4 mL), and water (1 mL). The mixture was stirred at room 30 temperature for 1 h and concentrated under reduced re. The residue was purified by reverse-phase prep-HPLC to afford 287 as a yellow solid (24 mg, 43%). MS-ESI: [M+H]+ 568.9. 1H NMR (500 MHz, CDCl3) δ 8.49 (d, J = 5.0 Hz, 1H), 7.99 (s, 1H), 7.72 (d, J = 2.0 Hz, 1H), 7.44 (s, 1H), 7.36 (d, J = 5.0 Hz, 1H), 6.91 (s, 1H), 5.74 (s, 1H), 5.04-5.02 (m, 1H), 4.64-4.62 (m, 1H), 4.56-4.54 (m, 1H), 4.39-4.37 (m, 1H), 4.17-3.92 (m, 4H), 3.86-3.84 (m, 408 1H), 3.72 (s, 3H), 3.45-3.37 (m,1H), 3.17-3.14 (m, 1H), 2.89-2.81 (m, 1H), 2.64-2.58 (m, 4H), 2.48 (s, 3H), 1.93-1.89 (m, 2H), 1.81-1.80 (m, 2H), 1.46 (d, J = 6.5 Hz, 3H).
Example 288a 1-Methyl(5-methylisoxazolylamino)(4,4,5,5- tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 288a 5 A 50-mL round bottomed flask equipped with a magnetic r and a reflux condenser was charged with 5-bromomethyl(5-methylisoxazolylamino)pyridin- 2(1H)-one 283a (330 mg, 1.16 mmol), Pin2B2 (442 mg, 1.74 mmol), a)3 (53 mg, 0.058 mmol), X-Phos (55 mg, 0.116 mmol), potassium acetate (227 mg, 2.32 mmol), and dioxane 10 (20 mL). After three cycles of vacuum/N2 flush, the mixture was heated at 70oC for 2 h.
Analysis of the reaction mixture by LCMS showed complete conversion to the desired product. It was cooled to room ature and filtered. The filtrate was concentrated under reduced pressure and the residue was washed with petroleum ether to afford 288a (300 mg, 78%) as yellow solid. MS-ESI: [M+H]+ 332.3 15 Example 288b 2-{4,4-Dimethyloxothia-10,11- diazatricyclo[6.4.0.02,6]dodeca-1(8),2(6),11-trienyl}{1-methyl[(5-methyl-1,2- yl)amino]oxo-1,6-dihydro-pyridinyl}pyridinecarbaldehyde 288b A 50-mL round-bottomed flask equipped with a magnetic stirrer was charged with 4- chloro{4,4-dimethyloxothia-10,11-diazatricyclo[6.4.0.02,6]dodeca-1(8),2(6),11- 20 trienyl}pyridinecarbaldehyde 282i (72 mg, 0.20 mmol), 288a (102 mg, 0.30 mmol), dppf) (16 mg, 0.020 mmol), K3PO4 (85 mg, 0.40 mmol), sodium e (33 mg, 0.40 mmol), acetonitrile (10 mL), and water (0.5 mL). After ng nitrogen into the mixture for 10 minutes, a reflux condenser was attached to the flask and the mixture was heated at 100 oC for 1 h. It was then cooled to room temperature and filtered. The filtrate was concentrated 25 under reduced pressure to afford 288b, which was used in the next step without further purification. MS-ESI: [M+H]+ 529.3. 409 Example 288 3-[3-(hydroxymethyl)[1-methyl[(5-methylisoxazolyl)amino]- 6-oxopyridyl]pyridyl]-7,7-dimethyl-6,8-dihydrocyclopenta[3,4]thieno[1,3-d]pyridazin- 4-one 288 A mixture of 288a (82 mg, 0.16 mmol) and NaBH4 (18.1 mg, 0.48 mmol) in methanol 5 (10 mL) was stirred at room temperature for 30 min. The mixture was quenched with water (5 mL) and evaporated under reduced pressure. The residue was extracted with dichloromethane (3 X 10 mL). The ed extract was trated under reduced pressure and the e was ed by reverse-phase prep-HPLC to afford 288 (54 mg, two steps: 34%) as white solid. MS-ESI: [M+H]+ 531.3. 1H NMR (500 MHz, DMSO-d6) δ 9.0 (s, 10 1H), 8.57 (d, J = 5.0 Hz, 1H), 8.46 (s, 1H), 8.00 (d, J = 1.5 Hz, 1H), 7.57 (d, J = 2.0 Hz, 1H), 7.51 (d, J = 5.0 Hz, 1H), 6.25 (s, 1H), 4.86 (bs, 1H), 4.39 (d, J = 8.5 Hz, 2H), 3.60 (s, 3H), 2.91 (s, 2H), 2.81 (s, 2H), 2.31 (s, 3H), 1.28 (s, 6H).
Example 289a tert-Butyl 3-Amino-1H-pyrazolecarboxylate 289a 15 To a mixture of 3-cyclopropyl-1H-pyrazolamine (3.0 g, 36 mmol) and triethylamine (7.6 g, 75 mmol) in 1,4-dioxane (35 mL) was added (Boc)2O (7.8 g, 36 mmol).
The on mixture was stirred at 25oC for 2 h. It was then concentrated under reduced pressure. The residue was purified by silica-gel column eluting with 3:1 petroleum ether/ethyl acetate to afford 289a as a white solid (3.4 g, 52%). MS-ESI: [M+H]+ 184.1. 20 Example 289b 3-(1H-Pyrazolylamino)bromomethylpyridin-2(1H)- one 289b 410 A 250-mL single-neck round-bottomed flask ed with a magnetic stirrer and a reflux condenser was charged with 289a (2.2 g, 12 mmol), XantPhos (0.69 g, 1.2 mmol), Pd2(dba)3 (1.1 g, 1.2 mmol), bromomethylpyridin-2(1H)-one (6.4 g, 24 mmol), Cs2CO3 (15.6 g, 48 mmol), and 1,4-dioxane (50 mL). After bubbling nitrogen through the 5 resulting mixture for 10 minutes, it was heated at 105oC for 15 h. The mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the residue the mixture was washed with ol (8 mL) to afford 289b as a pale yellow solid (1.2 g, 37%). MS-ESI: [M+H]+ 269.1.
Example 289c (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 10 2(6),7-dienyl}{1-methyloxo[(1H-pyrazolyl)amino]-1,6-dihydropyridin yl}pyridinyl)methyl Acetate 289c A 50-mL round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 289b (200 mg, 0.74 mmol), (2-{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-{3-[(acetoxy)methyl]{4,4-dimethyloxo-1,10- 15 diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (360 mg, 0.90 mmol), PdCl2(dppf) (41 mg, 0.050 mmol), K3PO4 (320 mg, 1.5 mmol), sodium acetate (123 mg, 1.5 mmol), acetonitrile (10 mL), and water (0.2 mL). The system was evacuated and then refilled with N2. Then it was heated at 90oC for 4 h. The mixture was cooled to room ature and filtered. The te was concentrated under reduced pressure and the residue 20 was purified by silica-gel column tography eluting with 10:1 dichloromethane/methanol to afford 289c as a pale yellow solid (150 mg, 38%). MS-ESI: [M+H]+ 542.3.
Example 289 3-[3-(hydroxymethyl)[1-methyloxo(1H-pyrazolylamino) pyridyl]pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazin 25 one 289 A mixture of 289c (150 mg, 0.28 mmol) and lithium hydroxide hydrate (236 mg, 5.6 mmol) in THF (4 mL), i-propanol (4 mL) and water (2 mL) was stirred at 40oC for 0.5 h. The mixture was ated under reduced pressure and d with water (10 mL). It was then extracted with ethyl acetate (3 X 15 mL). The combined extract was concentrated under 30 reduced pressure and the residue was purified by reverse-phase PLC to afford 289 as a pale yellow solid (25 mg, 18%). MS-ESI: [M+H]+ 499.9. 1H NMR (500 MHz, DMSO-d6) δ 12.07 (s, 1H), 8.47 (d, J = 5.0 Hz, 1H), 8.17 (s, 1H), 8.08 (s, 1H), 7.55-7.54 (m, 1H), 7.40- 7.39 (m, 1H), 7.32 (d, J = 5.0 Hz, 1H), 6.55 (s, 1H), 6.12 (s, 1H), 4.94-4.92 (m, 1H), 4.48- 411 4.47 (m, 1H), .39 (m, 1H), 4.23-4.17 (m, 3H), 3.84-3.82 (m, 1H), 3.59 (s, 3H), 2.58- 2.56 (m, 2H), 2.50-2.42 (m, 2H), 1.22 (s, 6H).
Example 290a 5-Bromomethyl(1-methyl-1H-pyrazolylamino)pyridin- 2(1H)-one 290a 5 A 250-mL single-neck bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 1,4-dioxane (100 mL), 1-methyl-1H-pyrazolamine (970 mg, 10.0 mmol), 3,5-dibromomethylpyridin(1H)-one (2.9 g, 11 mmol), and cesium carbonate (6.5 g, 20.0 mmol). After bubbling nitrogen through the suspension for 10 10 s, tris(dibenzylideneacetone)dipalladium(0) (457 mg, 0.50 mmol) and Xantphos (587 mg, 1.0 mmol) were added. The system was subjected to three cycles of vacuum/argon flush and heated at reflux for 2 h. It was then cooled to room ature and filtered. The solid was washed with dichloromethane (2 X 50 mL) and the combined organic te was concentrated. The residue was purified by silica-gel column chromatography eluting with 15 30:1 dichloromethane/methanol to afford 290a as a yellow solid (900 mg, 32%). MS-ESI: [M+H]+ 283.1 Example 290b (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{1-methyl[(1-methyl-1H-pyrazolyl)amino]oxo-1,6- opyridinyl}pyridinyl)methyl Acetate 290b 20 A 50-mL single-neck round-bottomed flask equipped with a ic stirrer and a reflux condenser was charged with {3-[(acetyloxy)methyl]{4,4-dimethyloxo-1,10- diazatri-cyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (595 mg, 1.5 mmol), 290a (282 mg, 1.0 mmol), K3PO4 (424 mg, 2.0 mmol), sodium acetate (164 mg, 2.0 mmol), 1,1’-bis(diphenylphosphino)ferrocenedichloropalladium(II) (82 mg, 0.1 mmol), 25 and acetonitrile/water(15/1 mL). After three cycles of vacuum/N2 flush, the mixture was heated at 100°C for 1.5 h. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was partitioned between dichloromethane (30 mL) and water (30 mL). The aqueous layer was extracted with dichloromethane (3 X 20 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced 412 pressure. The dark e was purified by silica-gel column chromatography eluting with dichloromethane/methanol (80/1 to 50/1) to 290b (300 mg, 54%) as yellow solid. MS-ESI: [M+H]+ 556.1 Example 290 3-[3-(hydroxymethyl)[1-methyl[(1-methylpyrazolyl)amino] 5 oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- b]pyrazinone 290 To a solution of 290b (139 mg, 0.25 mmol) in THF (5mL), propanol (5 mL), and water (2 mL) was added lithium hydroxide (60 mg, 2.5 mmol). The reaction mixture was stirred at room temperature for 2.5 h. It was then concentrated under reduced pressure. The 10 residue was partitioned n dichloromethane (20 mL) and water (10 mL). The aqueous layer was extracted with dichloromethane (3 X 10 mL). The combined extract was washed with brine, dried over , filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase prep-HPLC to afford 290 (30 mg, 23%) as white solid. MSESI : [M+H]+ 514.3. 1H NMR (500 MHz, DMSO-d6) δ 8.47 (d, J = 5.0 Hz, 1H), 8.16 (s, 1H), 15 8.03 (d, J = 2.5 Hz, 1H), 7.48 (d, J = 2.0 Hz, 1H), 7.39 (d, J = 1.5 Hz, 1H), 7.33 (d, J = 5.0 Hz, 1H), 6.55 (s, 1H), 6.07 (d, J = 2.0 Hz, 1H), 4.97 (t, J = 5.0 Hz, 1H), 4.47-4.40 (m, 2H), 4.24-4.18 (m, 3H), 3.85-3.83 (m, 1H), 3.70 (s, 3H), 3.58 (s, 3H), 2.58-2.56 (m, 2H), 2.42 (s, 2H), 1.22 (s, 6H).
Example 291a (4-{1-Methyl[(5-methyl-1,2-oxazolyl)amino]oxo-1,6- 20 dihydropyridinyl}{6-oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trien yl}pyridinyl)methyl Acetate 291a A 50-mL bottomed flask equipped with a reflux condenser was charged with oxothia-4,5-diazatricyclo[7.4.0.02,7]trideca-1(9),2(7),3-trienyl}(tetramethyl- 25 1,3,2-dioxaborolanyl)pyridinyl)methyl acetate 230i (150 mg, 0.31 mmol), 5-bromo methyl(5-methylisoxazolylamino)pyridine-2(1H)-one 283a (88 mg, 0.31 mmol), PdCl2(dppf) (24 mg, 0.031 mmol), K3PO4 (131 mg, 0.62 mmol), sodium acetate (61 mg, 0.62 mmol), water (0.2 mL), and acetonitrile (10 mL). The system was subjected to three cycles of /argon flush and stirred at 100°C for 3 h. The reaction mixture was cooled to room 413 temperature and filtered. The te was concentrated under reduced re. The residue was partitioned between dichloromethane (20 mL) and water (10 mL). The organic layer was separated and the water layer was extracted with dichloromethane (2 × 20 mL). The ed organic layer was dried over Na2SO4, ed, and concentrated under reduced 5 pressure. The dark residue was purified by -gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 291a (104 mg, 60%) as a yellow solid. MS-ESI: [M+H]+ 559.1 Example 291 3-[3-(hydroxymethyl)[1-methyl[(5-methylisoxazolyl)amino]- 6-oxopyridyl]pyridyl]-6,7,8,9-tetrahydrobenzothiopheno[2,3-d]pyridazinone 291 10 To a solution of 291a (100 mg, 0.18 mmol) in THF/ i-propanol /water (10/5/5 mL) was added lithium hydroxide (43 mg, 1.8 mmol) at room temperature. After being stirred for 1 h, MS indicated the reaction was complete. Then the mixture was concentrated under reduced pressure and the residue was partitioned between water (10 mL) and dichloromethane (15 mL). The water phase was extracted with dichloromethane (3 X 10 mL). 15 The combined organic layer was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase prep-HPLC to afford 291 (56 mg, 60%) as white solid. MS-ESI: [M+H]+ 517.2. 1H NMR (500 MHz, DMSO-d6) δ 9.02 (s, 1H), 8.58 (d, J = 5.0 Hz, 1H), 8.48 (s, 1H), 8.00 (d, J = 2.0 Hz, 1H), 7.58 (d, J = 2.0 Hz, 1H), 7.52 (d, J = 5.0 Hz, 1H), 6.25 (s, 1H), 4.87-4.85 (m, 1H), .36 20 (m, 2H), 3.61 (s, 3H), 2.98-2.85 (m, 4H), 2.32 (s, 3H), 1.92-1.86 (m, 4H).
Example 292a 4-{5-[(1,5-Dimethyl-1H-pyrazolyl)amino]methyloxo- 1,6-dihydro- pyridinyl}{4,4-dimethyloxothia-10,11- diazatricyclo[6.4.0.02,6]dodeca-1(8),(6),11- 10-yl}pyridinecarbaldehyde 292a N N NH O O N S N N O N 292a 25 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 3-(1,5-dimethyl-1H-pyrazolylamino)methyl (4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 242a (344 mg, 1.0 mmol), 4- chloro{4,4-dimethyloxothia-10,11-diazatricyclo[6.4.0.02,6]dodeca- 1(8),2(6),11- 414 trienyl}pyridinecarbaldehyde 282i (538.5 mg, 1.5 mmol), Pd2(dba)3 (91.5 mg, 0.10 mmol), tricyclohexylphospine (112 mg, 0.40 mmol), cesium carbonate (652 mg, 2.0 mmol), 1,4-dioxane (20 mL), and water (0.5 mL). After three cycles of vacuum/argon flush, the mixture was heated at 75oC for 2 h. It was then filtered and the te was evaporated under 5 reduced pressure. The residue was washed with petroleum ether to afford 292a (300 mg, crude) as a black solid. MS-ESI: [M+H]+ 542.2 Example 292 3-[4-[5-[(1,5-dimethylpyrazolyl)amino]methyloxopyridyl]- 3-(hydroxymethyl)pyridyl]-7,7-dimethyl-6,8-dihydrocyclopenta[3,4]thieno[1,3- d]pyridazinone 292 10 To a solution of 292a (162.6 mg, 0.30 mmol) in ol (6 mL) was added sodium borohydride (114 mg, 3.0 mmol) at 0oC. The reaction was stirred at 25oC for 0.5 h. It was then quenched with water (10 mL). The resulting mixture was evaporated under reduced re and the residue was extracted with dichloromethane (3 x 20 mL). The combined dichloromethane extract was concentrated under reduced pressure and the residue was 15 purified by e-phase prep-HPLC to afford 292 (35 mg, 22%) as a yellow solid. MS-ESI: [M+H]+ 543.8. 1H NMR (500 MHz, DMSO-d6) δ 8.57 (d, J = 5.0 Hz, 1H), 8.47 (s, 1H), 8.08 (s, 1H), 8.02 (d, J = 1.5 Hz, 1H), 7.52 (d, J = 5.0 Hz, 1H), 7.40 (d, J = 1.5 Hz, 1H), 5.90 (s, 1H), 4.88 (s, 1H), 4.40 (d, J = 5.0 Hz, 2H), 3.59 (s, 3H), 3.58 (s, 3H), 2.92 (d, J = 4.5 Hz, 2H), 2.81 (s, 2H), 2.18 (s, 3H), 1.29 (s, 3H), 1.28 (s, 6H). 20 Example 293a ylnitro-1H-1,2,3-triazole 293a Br O N N N Pd/C, MeOH, H2 Br N NH N rt, 4 h N O N N N NO2 N NH2 Pd2(dba)3,xantphos, N Br Cs2CO3, dioxane, 293b reflux, 18 h 293a 293c N N N NH 199e AcO O N N N Pd(dppf)Cl2, CH3CN, H2O, O N NaOAc, K3PO4, reflux, 2 h 293d 415 To a 100-mL single-neck round-bottomed containing 4-nitro-2H-1,2,3-triazole (2.0 g, 17.5 mmol) and THF (10 mL) at 0oC was added NaH (1.7 g, 35.0 mmol, 2.0 eq.). The e was stirred at 0oC for 15 min. A on of iodomethane (3.68 g, 26.3 mmol, 1.5 eq.) in acetone (40 mL) was added and the resulting reaction mixture was stirred at room 5 temperature for 2 h. After this time, the reaction was quenched by water (20 mL) at 0oC and concentrated under reduced pressure. The residue was diluted with dichloromethane (100 mL). It was then washed with brine, dried over ous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica-gel column chromatography g with 6:1 petroleum ethyl acetate to afford 293a (800 mg, 35%) as a light yellow solid and the 10 regioisomer 1-methylnitro-1H-1,2,3-triazole (1.34 g, 60%) as a white solid. 1H NMR (500 MHz, CDCl3) δ 8.34 (s, 1H), 4.26 (s, 3H).
Example 293b 1-Methyl-1H-1,2,3-triazolamine 293b Following the procedure in Example 130b, and starting with 293a (800 mg, 6.25 mmol) and 10% palladium on carbon (50% wet, 160 mg) afforded 293b as a yellow solid (600 15 mg, 98%). 1H NMR (500 MHz, CDCl3) δ 6.91 (s, 1H), 3.97 (s, 3H), 3.65 (brs, 2H).
Example 293c 5-Bromomethyl(1-methyl-1H-1,2,3-triazol o)pyridin-2(1H)-one 293c Following the procedure in Example 130c, and starting with 293b (500 mg, 5.10 mmol, 1.0 eq.) and 3,5-dibromomethylpyridin-2(1H)-one (2.04 g, 7.65 mmol, 1.5 eq.) 20 afforded 293c as a yellow solid (760 mg, 52%). MS-ESI: [M+H]+ 283.9.
Example 293d (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{1-methyl[(1-methyl-1H-1,2,3-triazolyl)amino]oxo-1,6- dihydropyridinyl}pyridinyl)methyl Acetate 293d ing the procedure in Example 283b, and starting with 293c (150 mg, 0.53 25 mmol, 1.0 eq.) and {3-[(acetoxy)methyl]{4,4-dimethyloxo-1,10 - diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (629 mg, 1.59 mmol, 3.0 eq.) afforded 293d as a yellow solid (110 mg, 37%). MS-ESI: [M+H]+ 557.4.
Example 293 3-[3-(hydroxymethyl)[1-methyl[(1-methyltriazolyl)amino] oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- 30 b]pyrazinone 293 Following the procedure in Example 283, and starting with 293d (110 mg, 0.20 mmol) afforded 293 as a pale yellow solid (78 mg, 75%). MS-ESI: [M+H]+ 514.9. 1H NMR (500 MHz, CDCl3) δ 8.49 (d, J = 4.5 Hz, 1H), 7.78 (s, 1H), 7.73 (s, 1H), 7.60 (s, 1H), 7.42 (s, 416 1H), 7.33 (d, J = 4.0 Hz, 1H), 6.87 (s, 1H), 5.25 (brs, 1H), 4.65-4.38 (m, 3H), 4.21-4.20 (m, 2H), 4.08 (s, 3H), 3.89-3.85 (m, 1H), 3.73 (s, 3H), 2.59 (s, 2H), 2.54 (s, 2H), 1.29 (s, 6H).
Example 294 3-[4-[5-[(5-tert-butylisoxazolyl)amino]methyloxopyridyl]- 3-(hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- 5 b]pyrazinone 294 Following the procedures in Example 273, and substituting 5-(tert-butyl)isoxazol amine for 2-aminopyridine, 294 was prepared (5.1 mg, 16% yield). 1H NMR (400 MHz, DMSO-d6) δ 8.93 (s, 1H), 8.48 (d, J = 5.0 Hz, 1H), 8.04 (d, J = 2.3 Hz, 1H), 7.56 (d, J = 2.3 Hz, 1H), 7.32 (d, J = 5.0 Hz, 1H), 6.56 (s, 1H), 6.22 (s, 1H), 4.90 (t, J = 5.3 Hz, 1H), 4.51 – 10 4.36 (m, 2H), 4.26 – 4.16 (m, 3H), 3.85 (d, J = 10.7 Hz, 1H), 3.60 (s, 3H), 2.58 (d, J = 7.7 Hz, 2H), 2.43 (s, 2H), 1.27 (s, 9H), 1.22 (s, 6H). ES-MS m/z 557.4 [M+1].
Example 295a 5-Ethylisoxazolamine 295a O N NH AcO O 273a N O N N N NH2 Pd2(dba)3 O N Xantphos 295a Cs2CO3 295b dioxane 100 oC, 3 h To a solution of 3-oxopentanenitrile (1.0 g, 10.3 mmol) in water (20 mL) was added 15 NaOH (535.6 mg, 13.4 mmol). After stirring for 5 s, ylamine hydrochloride (787.4 mg, 11.33 mmol) was added and mixture was heated at 40 oC for 12 h. At this point, conc. HCl (3 mL) was added and the reaction mixture was heated at 50 oC for 2 h. Analysis of the on e by LCMS showed complete conversion to the desired product. It was then cooled to room temperature and adjusted the pH to 10 with aqueous NaOH (30%). The 20 mixture was extracted with ethyl acetate (3 X 50 mL). The combined organic layer was washed with brine, dried over Na2SO4, and trated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 2:1 petroleum ether/ethyl acetate to afford 295a as a yellow solid (300 mg, 25%). MS-ESI: [M+H]+ 113.3. 1H NMR (500 MHz, DMSO-d6) 5.55 (s, 1H), 5.40 (s, 2H), 2.56-2.52 (m, 2H), 1.13 (t, J = 7.5 Hz, 3H). 25 Example 295b 4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{5-[(5-ethyl-1,2-oxazolyl)amino]methyloxo-1,6- dihydropyridinyl}pyridinyl)methyl Acetate 295b 417 A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 295a (24.8 mg, 0.222 mmol), [4-(5-bromomethyl 6-dihydropyridinyl){4,4-dimethyloxo-1,10-diazatri-cyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}pyridinyl]methyl e 273a (100 mg, 0.185 mmol), Pd2(dba)3 (8.5 mg, 5 0.0093 mmol), Xantphos (10.7 mg, 0.019 mmol), Cs2CO3 (120.6 mg, 0.37 mmol), and dioxane (10 mL). The system was subjected to three cycles of vacuum/nitrogen flush and heated at 100oC under N2 protection for 3 h. Analysis of the reaction mixture by LCMS showed te conversion to the desired product. It was cooled to room temperature and filtered. The te was concentrated under reduced pressure. The residue was washed with 10 acetonitrile (0.5 mL) to afford 295b as white solid (52 mg, 49.5%), which was used in the next step without further purification. MS-ESI: [M+H]+ 570.8 Example 295 3-[4-[5-[(5-ethylisoxazolyl)amino]methyloxopyridyl] (hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- b]pyrazinone 295 15 To a solution of 295b (42 mg, 0.0736 mmol) in THF (4 mL), i-propanol(4 mL), and water (4 mL) was added lithium hydroxide (17.7 mg, 0.736 mmol). The reaction mixture was stirred at room temperature for 1 h and concentrated under reduced pressure. The e was diluted with water (5 mL) and extracted with dichloromethane (3 X 20 mL). The combined extract was dried over Na2SO4, filtered, and concentrated under d pressure. The 20 residue was purified by reverse-phase prep-HPLC to afford 295 (22 mg, 57%) as a white solid. MS-ESI: [M+H]+ 528.8. 1H NMR (500 MHz, 6) δ 8.99 (s, 1H), 8.49 (d, J = 5.5 Hz, 1H), 8.03 (d, J = 2.0 Hz, 1H), 7.56 (d, J = 2.0 Hz, 1H), 7.33 (d, J = 5.0 Hz, 1H), 6.57 (s, 1H), 6.25 (s, 1H), 4.94-4.92 (m, 1H), 4.48-4.38 (m, 2H), 4.26-4.19 (m, 3H), 3.87-3.85 (m, 1H), 3.61 (s, 3H), 2.68-2.66 (m, 2H), 2.62-2.59 (m, 2H), 2.43 (s, 2H), 1.22 (s, 6H), 1.19 (t, J 25 = 7.5 Hz, 3H).
Example 296a (3-Nitro-1H-pyrazolyl)methanol 296a 418 Cl HO Br HO Br SOCl2 N HN N Cs2CO3, DMF Br N N NO2 r.t., 4 h Br N HCCl3, 70 °C, 3h NO2 NO2 296a 296b 296c O Br O O O N N N Br Pd/C, H2 NH2 N N EtOH, r.t., 2h DMSO, M.W. N Pd2(dba)3, xantphos N NH2 NO2 , dioxane 120 °C, 1h 100 °C, 5h 296d 296e O O N N N N N N NH NH 199e AcO O O N f)Cl2, K3PO4, N N N Br NaOAc, CH3CN/H2O, 100 °C, 1h O N 296f 296g A 100-mL three-neck round-bottomed flask equipped with a nitrogen inlet was purged with nitrogen and charged with 3-nitropyrazolecarboxylic acid (0.56 g, 3.56 mmol) and THF (8 mL). The system was cooled to -5 oC using an ice/acetone bath. Borane-THF 5 complex solution (1.0M, 11 mL, 11.0 mmol) was added at a rate that maintained the internal reaction temperature below 5 oC. After the addition was complete the g bath was removed and the reaction was d at 60 oC for 3 h. After this time the reaction was cooled to -5oC using an ice/acetone bath, water (2 mL) and 4N hydrochloric acid (2 mL) was added.
The reaction mixture was stirred at 70oC for 1 h in order to destroy the borane-pyrazole 10 complex. It was cooled to room temperature and concentrated under reduced pressure to a volume of approximately 1 mL. Ethyl acetate (20 mL) and water (10 mL) were added and the mixture was stirred for 15 min. The aqueous layer was separated and extracted with ethyl acetate (4 x 10 mL). The combined c layer was washed with saturated aqueous sodium 419 bicarbonate (2 x 10 mL), brine (10 mL), and dried over sodium sulfate. The drying agent was removed by filtration and the te was concentrated under reduced re to afford 296a (345 mg, 68%) as a light yellow solid. MS-ESI: [M+H]+ 144 Example 296b (1-(2-Bromoethyl)nitro-1H-pyrazolyl)methanol 296b 5 A mixture of 296a (345 mg, 2.41 mmol), and cesium carbonate (965 mg, 2.96 mmol) in DMF (5 mL) was cooled to 0oC using an ice/acetone bath and dibromoethane (4.48 g, 24.1 mmol) was added n-wise (no rm). The reaction was stirred at 0 oC for 1 h and room temperature for 4 h. After this time ethyl acetate (20 mL) and water (15 mL) were added. The aqueous layer was separated and extracted with ethyl acetate (2 x 10 mL). The 10 combined organic layer was washed with water (10 mL), brine (10 mL), and dried over sodium sulfate. The drying agent was removed by filtration. The filtrate was concentrated under reduced pressure to afford the crude product, which was purified by silica-gel column chromatography eluting with 6:1 petroleum ether/ethyl acetate to afford 296b (300 mg, 50%).
MS-ESI: [M+H]+ 250 15 Example 296c 1-(2-Bromoethyl)(chloromethyl)nitro-1H-pyrazole 296c A 50-mL three-necked bottomed flask equipped with a nitrogen inlet and a reflux condenser was purged with nitrogen and charged with 296b (438 mg, 1.76 mmol) and chloroform (10 mL). The reaction was cooled to -5 oC using an ice/acetone bath and SOCl2 (628 mg, 5.28 mmol) was added portion-wise. The g bath was removed and the 20 reaction was stirred at 70 oC for 3 h. After this time, the solvent was removed under reduced pressure. ethyl acetate was added to the residue and the resulting solution was cooled to -5oC.
Saturated aqueous sodium bicarbonate (3 mL) was added until a pH of 8.5 was reached. The mixture was partitioned between ethyl acetate and water. The combined organic layer was washed with saturated aqueous sodium ate (2 x 5 mL), brine (10 mL), and dried over 25 sodium sulfate. The drying agent was removed by filtration. The filtrate was concentrated under reduced pressure to afford 296c (284 mg, 60%).
Example 296d 5-(2-Methoxyethyl)nitro-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazine 296d A microwave vial equipped with a magnetic stirrer was charged with 296c (2.67 g, 30 10.0 mmol), 2-methoxyethanamine (2.25 g, 30.0 mmol), and DMSO (14 mL). The reaction mixture was heated at 120ºC under microwave irradiation for 1.0 h. It was cooled to room temperature and diluted with ethyl acetate (40 mL). The e was washed with water (3 X 15 mL). The organic layer was dried and filtered. The te was concentrated under pressure and the al was purified by silica-gel column chromatography eluting with 30:1 420 dichloromethane/methanol to afford 296d (1.7 g, 75%) as a yellow solid. MS-ESI: [M+H]+ 227.0 Example 296e 5-(2-Methoxyethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin- 2-amine 296e 5 To a solution of 296d (1.7 g, 7.5 mmol) in ethanol (50 mL) was added Pd/C (10%, 800 mg). The reaction was charged with hydrogen gas (via balloon) and stirred at room temperature for 2 h. After reaction was complete, the e was filtered through a plug of ®. The filtrate was concentrated reduced pressure to afford 296e as a yellow solid (1.2 g, 82%), which was used directly without further purification. MS-ESI: [M+H]+ 197.3 10 Example 296f 5-Bromo(5-(2-methoxyethyl)-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)methylpyridin-2(1H)-one 296f A 100-mL -neck round-bottomed flask equipped with a magnetic r and a reflux condenser was charged with 1,4-dioxane (40 mL), 296e (588 mg, 3.0 mmol), 3,5- dibromomethylpyridin-2(1H)-one (800 mg, 3.0 mmol), and cesium ate (1.96 g, 6.0 15 mmol). After bubbling nitrogen through the suspension for 20 minutes, xantphos (173 mg, 0.30 mmol) and ibenzylideneacetone)dipalladium(0) (137 mg, 0.15 mmol) were added.
The system was subjected to three cycles of vacuum/argon flush and heated at reflux for 5 h.
It was then cooled to room temperature and filtered. The solid was washed with dichloromethane (2 X 15 mL). The combined filtrate was concentrated under reduced 20 pressure. The residue solid was purified by silica-gel column chromatography eluting with dichloromethane/methanol (80/1 to 30/1) to afford 296f (745 mg, 65%) as yellow solid. MSESI : [M+H]+ 382.9 e 296g 4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}(5-{[5-(2-methoxyethyl)-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin 25 yl]amino}methyloxo-1,6-dihydropyridinyl)pyridinyl)methyl Acetate 296g A 25-mL round-bottomed flask equipped with a reflux condenser was charged with {3-[(acetoxy)methyl]{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- dienyl}pyridinyl}boronic acid 199e (198 mg, 0.50 mmol), 296f (190 mg, 0.50 mmol), K3PO4 (212 mg, 1.0 mmol), sodium acetate (82 mg, 1.0 mmol), Pd(dppf)Cl2 (21 mg, 0.025 30 mmol), acetonitrile (8 mL), and water (0.5 mL). The system was ted to three cycles of vacuum/nitrogen flush and heated at 100 oC under N2 protection for 1 h. Analysis of the reaction mixture by LCMS showed complete conversion to the desired product. The on mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue was partitioned between dichloromethane (20 mL) and water 421 (10 mL). The water layer was extracted with dichloromethane (2 × 20 mL). The combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure.
The dark residue was ed by silica-gel column tography eluting with dichloromethane/methanol (80/1 to 30/1) to afford 296g (163 mg, 50%) as a yellow solid. 5 MS-ESI: [M+H]+ 654.9 Example 296 3-[3-(hydroxymethyl)[5-[[5-(2-methoxyethyl)-6,7-dihydro-4H- pyrazolo[1,5-a]pyrazinyl]amino]methyloxopyridyl]pyridyl]-7,7-dimethyl- 1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 296 To a solution of 296g (160 mg, 0.245 mmol) in THF/i-propanol/water(8/5/3mL) was 10 added lithium hydroxide (29 mg, 1.22 mmol). The e was stirred at room temperature for 1 h and evaporated under pressure. The residue was purified by reverse-phase prep-HPLC to afford 296 as a white solid (117 mg, 78%). MS-ESI: [M+H]+ 613.3. 1H NMR (500 MHz, CDCl3) δ 8.48 (d, J = 5.0 Hz, 1H), 7.93 (d, J = 2.0 Hz, 1H), 7.73 (d, J = 2.5 Hz, 1H), 7.41 (s, 1H), 7.35 (d, J = 5.0 Hz, 1H), 6.86 (s, 1H), 5.71 (s, 1H), 5.05 (t, J = 7.0 Hz, 1H), 4.66-4.65 15 (m, 1H), 4.52-4.50 (m, 1H), 4.36-4.34 (m, 1H), 4.17-4.05 (m, 2H), 4.10-4.08 (m, 2H), 3.88- 3.87 (m, 1H), 3.75-3.73 (m, 2H), 3.71 (s, 3H), 3.61-3.59 (m, 2H), 2.40 (s, 3H), 3.04-3.03 (m, 2H), 2.80 (t, J = 5.5 Hz, 2H), 2.59-2.57 (m, 2H), .53 (m, 2H), 1.29 (s, 6H).
Example 297a 6-Chloromethyl(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5- zinylamino)pyridazin-3(2H)-one 297a 20 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 1,4-dioxane (30 mL), 5-methyl-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinamine 113e (1.70 g, 11.2 mmol), 4-bromochloro methylpyridazin-3(2H)-one (2.68 g, 12.0 mmol), and cesium carbonate (7.30 g, 22.4 mmol). 25 After ng en through the suspension for 30 minutes, Xantphos (0.59 g, 1.02 mmol) and tris(dibenzylideneacetone)dipalladium(0) (467 mg, 0.51 mmol) were added. The system was subjected to three cycles of vacuum/argon flash and heated at 90oC for 2 h. It was then cooled to room temperature and filtered. The filtrate was evaporated in vacuo. The 422 residue was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 297a (1.9 g, 60%) as a brown solid. LCMS: [M+H]+ 295.1 Example 297b (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 5 2(6),7-dienyl}[1-methyl({5-methyl-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin no)oxo-1,6-dihydropyridazinyl]pyridinyl)methyl Acetate 297b A 25-mL round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 297a (195 mg, 0.66 mmol), (2-{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}(tetramethyl-1,3,2-dioxaborolan 10 yl)pyridinyl)methyl acetate 199e (315 mg, 0.66 mmol), PdCl2(dppf) (40 mg, 0.050 mmol), K3PO4 (250 mg, 1.2 mmol), sodium acetate (100 mg, 1.20 mmol), itrile (8 mL), and water (1 mL). The system was evacuated and then ed with N2. It was then heated at 100oC for 1 h. The mixture was cooled to room temperature and filtered. The te was concentrated under reduced pressure and the e was purified by silica-gel column 15 chromatography eluting with 20:1 dichloromethane/methanol to afford 297b as a yellow solid (150 mg, 38%). MS-ESI: [M+H]+ 612.3.
Example 297 3-[3-(hydroxymethyl)[1-methyl[(5-methyl-6,7-dihydro-4H- lo[1,5-a]pyrazinyl)amino]oxo-pyridazinyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 297 20 A mixture of 297b (150 mg, 0.24 mmol) and lithium hydroxide hydrate (96 mg, 2.4 mmol) in THF (8 mL), i-propanol (8 mL), and water (2 mL) was stirred at 40oC for 0.5 h.
The mixture was evaporated under reduced re and the residue was partitioned between dichloromethane (15 mL) and water (10 mL). The combined extract was concentrated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 297 as a 25 pale yellow solid (98 mg, 70%). MS-ESI: [M+H]+ 570.3. 1H NMR (500 MHz, 6) δ 9.30 (s, 1H), 8.52 (d, J = 4.5 Hz, 1H), 7.90 (s, 1H), 7.38 (d, J = 5.0 Hz, 1H), 6.55 (s, 1H), 5.99 (s, 1H), 4.74-4.73 (m, 1H), 4.60-4.58 (m, 1H), 4.40-4.37 (m, 1H), 4.26-4.24 (m, 1H), 4.19-4.18 (m, 2H), 3.96-3.95 (m, 2H), 3.89-3.87 (m, 1H), 3.75 (s, 3H), 3.53-3.52 (m, 2H), 2.80-2.78 (m, 2H), 2.57-2.55 (m, 2H), 2.52-2.50 (m, 2H), 2.35 (s, 3H), 1.22 (s, 6H). 30 Example 298a (4-{5-[(5-Cyanopyrazinyl)amino]methyloxo-1,6- dihydropyridinyl}{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- dienyl}pyridinyl)methyl a\Acetate 298a 423 A 50-mL round-bottomed flask equipped with a reflux condenser was charged with bromomethyloxo-1,6-dihydropyridinyl){4,4-dimethyloxo-1,10- diazatricy-clo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl]methyl acetate 273a (269 mg, 5 0.50 mmol), 5-aminopyrazinecarbonitrile (60 mg, 0.50 mmol), XantPhos (29 mg, 0.050 mmol), Pd2(dba)3 (45 mg, 0.050 mmol), Cs2CO3 (326 mg, 1.0 mmol), and 1,4-dioxane (10 mL). The reaction mixture was heated at 100oC under microwave irradiation for 1h after three times atmosphere/argon flush. The mixture was filtered off and the solid was washed with methanol (50 mL). The combined filtrate was evaporated under reduced pressure and the 10 residue was purified with silica-gel column chromatography eluting with 50:1 dichloromethane/methanol to afford 298a (200 mg, 69%) as yellow solid. MS-ESI: [M+H]+ 579.3.
Example 298 [2-(7,7-dimethyloxo-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinyl)(hydroxymethyl)pyridyl] 15 methyloxopyridyl]amino]pyrazinecarbonitrile 298 A mixture of 298a (200 mg, 0.35 mmol) and lithium hydroxide (84 mg, 3.5 mmol) in i-propanol /THF (5 mL/5 mL) and water (2 mL) was stirred at room temperature for 2 h. The mixture was evaporated under reduced pressure and the residue was partitioned n ethyl e (20 mL) and water (10 mL). The combined extract was concentrated under reduced 20 pressure and the residue was purified by reverse-phase prep-HPLC to afford 298 (40 mg, 21%) as yellow solid. MS-ESI: [M+H]+ 537.3. 1H NMR (500 MHz, 6) δ 9.96 (s, 1H), 8.77 (s, 1H), 8.701 (d, J = 2.5 Hz, 1H), 8.67 (s, 1H), 8.50 (d, J = 5.0 Hz, 1H), 7.76 (d, J = 2.0 Hz, 1H), 7.35 (d, J = 5.0 Hz, 1H), 6.56 (s, 1H), 5.00 (t, J = 5.0 Hz, 1H), 4.42-4.39 (m, 2H), 4.24-4.19 (m, 3H), 3.84 (m, 1H), 3.63 (s, 3H), 2.57 (m, 2H), 2.42 (s, 2H), 1.23 (s, 6H) 25 Example 299a 5-Phenylisoxazolamine 299a 424 Br O O N O N NH Br N NH2 O Xantphos, Cs2CO3, Pd2(dba)3, 1,4-dioxane, N 299a Br 95 °C, 3 h 299b O N NH AcO O 199e N N N 0.1eq PdCl2(dppf), 2eq K3PO4 2eq CH3COONa, CH3CN, O N 100 °C, 3h, H2O 299c To a stirred solution of 3-oxophenylpropanenitrile (1.5 g, 10.3 mmol) and NaOH (452 mg, 11.3 mmol) in water (10 mL)/EtOH (10 mL) was added ylamine hydrochloride (785 mg, 11.3 mmol). The mixture was stirred at 80oC for overnight. At this 5 point, conc. HCl (1.3 mL, 15.5 mmol) was added and the resulting mixture was heated at 80oC for 2 h. It was then basified to pH 10 and extracted with ethyl acetate. The combined extract was concentrated under reduced pressure and the residue was purified by silica-gel column chromatography eluting with ethyl acetate/petroleum ether (1:50 to 1:10) to afford 299a as a yellow solid (1.1 g, 68%). MS-ESI: [M+H]+161.3. 10 Example 299b 5-Bromomethyl(5-phenylisoxazolylamino)pyridin- 2(1H)-one 299b A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 1,4-dioxane (40 mL), 299a (640 mg, 4.0 mmol), 3,5- omethylpyridin-2(1H)-one (1.1 g, 4.0 mmol), Pd2(dba)3 (366.8 mg, 0.40 mmol), 15 XantPhos (462.4 mg, 0.80 mmol), and cesium carbonate (2.6 g, 8.0 mmol). After three cycles of /argon flush, the mixture was heated at 92oC for 3 hrs. It was then cooled to room temperature and filtered. The filtrate was trated under reduced re and the resulting e was washed with acetonitrile to afford 299b (1.7 g, 87%). MS-ESI: [M+H]+ 346.0. 425 Example 299c (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{1-methyloxo[(5-phenyl-1,2-oxazolyl)amino]-1,6- dihydropyridin-pyridinyl}pyridinyl)methyl e 299c A 50-mL round-bottomed flask equipped with a magnetic stirrer and a reflux 5 condenser was charged with 299b (138 mg, 0.40 mmol), cetoxy)methyl]{4,4- dimethyloxo-1,10-diaza-tricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (158.8 mg, 0.40 mmol), Pd(dppf)Cl2 (32.7 mg, 0.040 mmol), K3PO4 (169.6 mg, 0.80 mmol), sodium acetate (108.8 mg, 0.80 mmol), water (0.5 mL), and acetonitrile (10 mL).
After three cycles of vacuum/argon flush, the mixture was heated at 100oC for 3 hrs. It was 10 then cooled to room temperature and filtered. The filtrate was concentrated under d pressure and the resulting residue was purified by silica-gel column chromatography eluting with 50:1 dichloromethane/methanol to afford 299c as a brown solid (120 mg, 49%). MSESI : [M+H]+ 618.8.
Example 299 3-[3-(hydroxymethyl)[1-methyloxo[(5-phenylisoxazol 15 yl)amino]pyridyl]pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- zinone 299 A mixture of 299c (100 mg, 0.16 mmol) and lithium hydroxide (96 mg, 4.0 mmol) in i-propanol /THF (1:1, 4 mL) and water (1 mL) was stirred at 40oC for 30 mins. The reaction mixture was concentrated under reduced pressure and diluted with water (5 mL). The 20 resulting mixture was extracted with dichloromethane for three times. The combined organic layer was concentrated under d pressure and the resulting residue was purified by reverse-phase prep-HPLC to afford 299 as a white solid (35 mg, 31%). MS-ESI: [M+H]+ 576.8. 1H NMR (500 MHz, DMSO-d6) δ 9.17 (s, 1H), 8.51 (d, J = 5.5 Hz, 1H), 8.11 (d, J = 2.5 Hz, 1H), .77 (m, 2H), 7.60 (d, J = 2.0 Hz, 1H), 7.55-7.49 (m, 3H), 7.35 (d, J = 4.5 25 Hz, 1H), 6.91 (s, 1H), 6.57 (s, 1H), 4.96-4.93 (m, 1H), .40 (m, 2H), 4.26-4.19 (m, 3H), 3.88-3.85 (m, 1H), 3.63 (s, 3H), 2.62-2.59 (m, 2H), 2.44-2.42 (m, 2H), 1.23 (s, 6H).
Example 300a 5-(1-Methoxypropanyl)nitro-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazine 300a 426 O O Br O N N N P Br d/ N EtOHC N Pd2(dba)3, xantphos N NO2, r.t., N NH2 Cs2CO3, e, 2h 100 °C, 5h 300a 300b O O N N N N NH N 199e AcO O N NH N O N N Pd(dppf)Cl2, N K3PO4, NaOAc, O N Br H2O, 100 °C, 1h 300d 300c A microwave vial ed with a magnetic stirrer was charged with 1-(2- bromoethyl)(chloromethyl)nitro-1H-pyrazole 296c (1.0 g, 3.7 mmol), 1- methoxypropanamine (1.0 g, 11.2 mmol), and DMSO (6 mL). The mixture was heated at 5 120 ºC under microwave irradiation for 1.0 h. It was the cooled to room temperature and diluted with water (30 mL). The resulting mixture was extracted with ethyl acetate (3 X 20 mL). The combined organic layer was dried and concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with petroleum ether/ethyl acetate(60/1 to 1/1) to afford 300a (600 mg, 68%) as a yellow solid. MS-ESI: [M+H]+ 241.0 10 Example 300b 5-(1-Methoxypropanyl)-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinamine 300b A solution of 300a (600 mg, 2.5 mmol) in EtOH (40 mL) was added Pd/C (10%, 60 mg). The reaction mixture was d with hydrogen gas (via n) and stirred at room temperature for 2 h. After reaction was complete, the mixture was filtered through a plug of 15 CELITE®. The filtrate was concentrated reduced pressure to afford 300b as a yellow solid (467 mg, 89%), which was used without r purification. MS-ESI: [M+H]+ 211.1 Example 300c 5-Bromo(5-(1-methoxypropanyl)-4,5,6,7- tetrahydropyrazolo[1,5-a]-pyrazinylamino)methylpyridin-2(1H)-one 300c 427 A 100-mL single-neck round-bottomed flask ed with a magnetic stirrer and a reflux condenser was charged with 1,4-dioxane (40 mL), 300b (400 mg, 1.9 mmol), 3,5- dibromomethylpyridin-2(1H)-one (H-001) (508 mg, 1.9 mmol), and cesium carbonate (1.24 g, 3.8 mmol). After bubbling nitrogen through the suspension for 20 minutes, xantphos 5 (109 mg, 0.19mmol) and Pd2(dba)3 (87 mg, 0.095 mmol) were added. The system was ted to three cycles of vacuum/argon flush and heated at reflux for 5 h. It was then cooled to room ature and filtered. The solid was washed with dichloromethane (2 X 30 mL). The combined filtrate was concentrated under reduced pressure. The residue was ed by silica-gel column chromatography eluting with romethane/methanol (80/1 10 to 30/1) to afford 300c (436 mg, 58%) as yellow solid. : [M+H]+ 396.0 Example 300d (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}(5-{[5-(1-methoxypropanyl)-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin- 2-yl]amino}methyloxo- 1,6-dihydropyridinyl)pyridinyl)methyl Acetate 300d A 50-mL round bottomed flask equipped with a reflux condenser was charged with 15 {3-[(acetyloxy)methyl]{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- dienyl}pyridinyl}boronic acid 199e (238 mg, 0.60 mmol), 300c(240 mg, 0.80 mmol), K3PO4 (254 mg, 1.2 mmol), sodium acetate (98 mg, 1.6 mmol), Pd(dppf)Cl2 (22 mg, 0.030 mmol), and acetonitrile/water (12/0.5 mL). The system was subjected to three cycles of vacuum/nitrogen flush and heated at 100oC under N2 tion for 1h. Analysis of the 20 reaction mixture by LCMS showed complete conversion to the desired product. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The e was partitioned between dichloromethane (30 mL) and water (30 mL). The water layer was extracted with dichloromethane (2 × 30 mL). The combined organic extract was dried over Na2SO4, ed, and concentrated under reduced pressure. 25 The dark residue was purified by silica-gel column tography eluting with dichloromethane/methanol (80/1 to 30/1) to afford 300d (200 mg, 50%) as yellow solid. MSESI : [M+H]+ 669.4 Example 300 (R)(3'-(hydroxymethyl)((5-(1-methoxypropanyl)-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinyl)amino)methyloxo-1,6-dihydro-[3,4'-bipyridin]- 30 2'-yl)-7,7-dimethyl-2,3,4,6,7,8-hexahydro-1H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazinone 300 To a solution of 300d (200 mg, 0.30 mmol) in THF/i-propanol/water(6/3/3mL) was added lithium hydroxide (36 mg, 1.5 mmol). The mixture was stirred at 30ºC for 1 h and concentrated under reduced pressure. The residue was partition between ethyl acetate (15 428 mL) and (10 mL). The water phase was ted with ethyl acetate (3 X 10 mL). The combined organic layer was dried, filtered, and concentrated under reduced pressure. Prep- HPLC and chiral resolution afforded the two enantiomers: 300 (35 mg, 18.6%) as white solid; and 303 (28 mg, 15 %) as white solid. : [M+H]+ 627.4. 1H NMR (500 MHz, CDCl 3) 5 δ 8.49 (d, J = 5.0 Hz, 1H), 7.92 (d, J = 2.5 Hz, 1H), 7.73 (d, J = 2.0 Hz, 1H), 7.41 (s, 1H), 7.35 (d, J = 5.0 Hz, 1H), 6.85 (s, 1H), 5.72 (s, 1H), 5.05-5.03 (m,1H), 4.67-4.45 (m,1H), 4.52-4.50 (m, 1H), 4.35-4.31 (m, 1H), .16 (m, 2H), 4.09-4.07 (m, 2H), .85 (m, 3H), 3.71 (s, 3H), 3.56-3.52 (m, 1H), 3.45-3.43 (m, 1H), 3.38 (s, 3H), 3.11-3.08 (m, 3H), 2.60-2.58 (m, 2H), 2.53 (s, 2H), 1.29 (s, 6H), 1.17 (d, J = 6.0 Hz, 3H). 10 Example 301a 6-(Trifluoromethyl)pyridazinamine 301a A mixture of 3-chlorotrifluoromethylpyridazine (1.6 g, 8.80 mmol) and ammonium hydroxide (9 mL) in THF (3 mL) was heated at 100 0C in a microwave reactor for 1 h. After this period, the reaction mixture was evaporated and the residue was extracted with 15 dichloromethane. The combined extract was dried over with MgSO4, filtered, and evaporated under reduce pressure to afford 301a (1.3 g, 93%) as a white solid. MS-ESI: [M+H]+ 164.1 Example 301b 5-Bromomethyl(6-(trifluoromethyl)pyridazin ylamino)pyridin-2(1H)-one 301b A 100-mL single-neck round-bottomed flask ed with a magnetic stirrer and a 20 reflux condenser was charged with 301a (750 mg, 4.6 mmol), XantPhos (532 mg, 0.92 mmol), Pd2dba3 (421 mg, 0.46 mmol), 2-bromochloronicotinaldehyde (H-001) (1.84 g, 6.9 mmol), Cs2CO3 (3.0 g, 9.2 mmol), and 1,4-dioxane (50 mL). The system was subjected to three cycles of vacuum/argon flush and heated at 90 oC for overnight. After the completion of the reaction, the mixture was filtered and the solid was washed with methanol (30 mL). The 25 ed filtrate was evaporated under reduced pressure and the residue was purified by silica-gel column tography eluting with 20:1 dichloromethane/methanol to afford 301b (1.38 g, 89%) as a yellow solid. MS-ESI: [M+H]+ 350.8 429 Example 301c (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}(1-methyloxo{[6-(trifluoromethyl)pyridazinyl]amino}-1,6- dihydropyridinyl)pyridinyl)methyl Acetate 301c A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 5 reflux condenser was charged with 301b (300 mg, 0.86 mmol), (2-{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}(tetramethyl-1,3,2-dioxaborolan yl)pyridinyl)methyl acetate 199e (824 mg, 1.72 mmol), CH3COONa (140 mg, 1.72 mmol), dppf) (70 mg, 0.086 mmol), K3PO4 (360 mg, 1.72 mmol), acetonitrile (20 mL), and water (0.5 mL). After bubbling nitrogen through the resulting mixture for 20 minutes, it was 10 heated at 100oC under nitrogen atmosphere for 2 h. The mixture was cooled to room temperature and filtered. The filtrate was ated under reduced pressure and the residue was purified by silica-gel column chromatography g with 50:1 dichloromethane/methanol to afford 301c as white solid (125 mg, 23%). MS-ESI: [M+H]+ 622.3 15 Example 301 3-[3-(hydroxymethyl)[1-methyloxo[[6- uoromethyl)pyridazinyl]amino]pyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 301 A mixture of 301c (90 mg, 0.14 mmol) and lithium hydroxide (24 mg, 0.56 mmol) in i-propanol /THF/water (6 mL /4 mL /2 mL) was stirred at room temperature for 0.5 h. The 20 mixture was evaporated under d pressure and the residue was portioned between dichloromethane (20 mL) and water (10 mL). The combined dichloromethane extract was concentrated under reduced pressure and the residue was purified by reverse-phase prep- HPLC to afford 301c as a white solid (39 mg, 48%). MS-ESI: [M+H]+ 580.3. 1H NMR (500 MHz, DMSO-d6) δ 9.57 (s, 1H), 8.87 (d, J = 2.5 Hz, 1H), 8.51 (d, J = 5.0 Hz, 1H), 7.96 (d, J 25 = 9.0 Hz, 1H), 7.88 (d, J = 9.0 Hz, 1H), 7.74 (d, J = 2.0 Hz, 1H), 7.38 (d, J = 5.0 Hz, 1H), 6.58 (s, 1H), 4.98-4.97 (m, 1H), 4.48-4.40 (m, 2H), 4.27-4.20 (m, 3H), 3.88-3.86 (m, 1H), 3.65 (s, 3H), 2.62-2.53 (m, 2H), 2.42-2.41 (m, 2H), 1.2 (s, 6H).
Example 302a N-Methyl(1-methylnitro-1H-pyrazolyl)methanamine 302a 30 To a d solution of MeNH2 (30% wt in water) (2.5 g, 20 mmol) in acetone (10 mL) at 0ºC (ice bath) was added K2CO3 (415 mg, 3 mmol), followed by the dropwise addition of a solution of 5-(bromomethyl)methylnitro-1H-pyrazole (220 mg, 1 mmol) in acetone (5 mL). The on mixture was then warmed to room temperature and stirred for 3 h. The solvent was removed and the e was ted with methylene chloride (3 X 15 mL), 430 dried over Na2SO4 and concentrated to afford 302a as a yellow oil (170 mg, 99%), which was used in the next step without additional purification. LCMS: (M+H)+ 171 Example 302b N-Methyl-N-((1-methylnitro-1H-pyrazol yl)methyl)oxetanamine 302b 5 To a mixture of 302a(170 mg, 1 mmol) in ol (4 mL), ZnCl2 (1 mmol/L in diethyl ether) (2 mL, 2 mmol) and one (150 mg, 2 mmol) were added at room temperature under en protection, followed by the addition of NaBH3CN (130 mg, 2 mmol). The reaction mixture was warmed to 50ºC and stirred for 3 h. The mixture was then cooled to room temperature and the solvent was removed. The residue was purified on flush 10 column eluting with 50:1 methylene chloride/methanol to afford 302b as a yellow solid (180 mg, 80%, two steps). LCMS: (M+H)+ 227. 1H NMR (500 MHz, DMSO) δ 6.99 (s, 1H), 4.52 (t, J=6.5, 2H), 4.42 (t, J=6, 2H), 3.98 (s, 3H), 3.63 (m, 1H), 3.50 (s, 2H), 2.03 (s, 3H). e 302c 1-Methyl((methyl(oxetanyl)amino)methyl)-1H-pyrazol amine 302c 15 To a solution of 302b (1.8 g, 7.96 mmol) in ethanol (20 mL) and water (20 mL), NH4Cl (3.3 g, 63.6 mmol) and iron powder (1.80 g, 31.8 mmol) were added. The reaction mixture was heated at 70ºC for 2 h. After that, the mixture was cooled to room ature and filtered. The filtrate was evaporated and the residue was extracted with methylene chloride (3 X 30 mL), dried Na2SO4, and concentrated to afford the crude product, which was 20 purified on flash column eluting with 50:1 methylene chloride/methanol containing 0.5% triethylamine to afford 302c as a yellow oil (1.3 g, 83%). LCMS: (M+H)+ 197 Example 302d omethyl(1-methyl((methyl(oxetan yl)amino)methyl)-1H-pyrazolylamino)pyridin-2(1H)-one 302d 25 Following the procedure in Example 292c, and starting with 302c and 3,5-dibromo methylpyridin-2(1H)-one afforded 302d in 63% yield. LCMS: (M+H)+ 383. 1H NMR (500 431 MHz, DMSO) δ 8.35 (s, 1H), 7.99 (d, J=2.5, 1H), 7.36 (d, J=2.5, 1H), 5.99 (s, 1H), 4.50 (t, J=7, 2H), 4.40 (t, J=6.5, 2H), 3.77 (s, 3H), 3.57 (m, 1H), 3.49 (s, 3H), 3.35 (s, 2H), 2.01 (s, 3H).
Example 302e (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 5 2(6),7-dienyl}{1-methyl[(1-methyl{[methyl(oxetanyl)amino]methyl}-1H- pyrazolyl)amino]oxo-1,6-dihydropyridinyl}pyridinyl)methyl e 302e A 25-mL round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 302d (200 mg, 0.52 mmol), {3-[(acetoxy)methyl]{4,4- dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dien yl}pyridinyl}boronic 10 acid 199e (210 mg, 0.53 mmol), PdCl2(dppf) (37 mg, 0.050 mmol), K3PO4 (250 mg, 1.2 mmol), sodium acetate (100 mg, 1.20 mmol), acetonitrile (8 mL), and water (0.5 mL). The system was evacuated and then ed with N2. The e was heated at 100 ºC for 1 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica-gel column chromatography eluting 15 with 20:1 dichloromethane/methanol to afford 302e as a yellow solid (150 mg, 44%). MSESI : [M+H]+ 655.3.
Example 302 3-[3-(hydroxymethyl)[1-methyl[[1-methyl[[methyl(oxetan yl)amino]methyl]pyrazolyl]amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 302 20 A mixture of 302e (150 mg, 0.23 mmol) and lithium hydroxide hydrate (90 mg, 2.3 mmol) in THF (8 mL), i-propanol (8 mL), and water (2 mL) was stirred at 40ºC for 0.5 h.
The mixture was concentrated under reduced pressure. The residue was partitioned between water (10 mL) and dichloromethane (3 X 15 mL). The ed organic t was concentrated under reduced pressure and the residue was purified by reverse-phase prep- 25 HPLC to afford 302 as a pale yellow solid (105 mg, 75%). MS-ESI: [M+H]+ 612.8. 1H NMR (500 MHz, DMSO-d6) δ 8.48 (d, J = 5.0 Hz, 1H), 8.14 (s, 1H), 8.05 (d, J = 2.5 Hz, 1H), 7.40 (d, J = 2.0 Hz, 1H), 7.33 (d, J = 5.0 Hz, 1H), 6.55 (s, 1H), 6.01 (s, 1H), 4.99-4.97 (m, 1H), 4.50-4.46 (m, 3H), 4.40-4.38 (m, 3H), 4.25-4.18 (m, 3H), 3.85-3.84 (m, 1H), 3.72 (s, 3H), 3.59 (s, 3H), 3.58-3.54 (m, 1H), 2.58-2.56 (m, 2H), 2.52-2.50 (m, 2H),2.43-2.42 (m, 2H), 30 2.00 (s, 3H), 1.22 (s, 6H).
Example 303 -[3-(hydroxymethyl)[5-({5-[(2R)methoxypropanyl]- 6H,7H-pyrazolo[1,5-a]pyrazinyl}amino)methyloxo-1,6-dihydropyridin yl]pyridinyl]-4,4-dimethyl-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienone 303 432 Following Example 300, single enantiomer 303 was obtained (28 mg, 15 %) as a white solid. MS-ESI: [M+H]+ 627.4. 1H NMR (500 MHz, CDCl 3) δ 8.49 (d, J = 5.0 Hz, 1H), 7.92 (d, J = 2.5 Hz, 1H), 7.73 (d, J = 2.0 Hz, 1H), 7.41 (s, 1H), 7.35 (d, J = 5.0 Hz, 1H), 6.85 (s, 1H), 5.72 (s, 1H), 5.05-5.03 (m,1H), 4.67-4.45 (m,1H), .50 (m, 1H), 4.35-4.31 (m, 5 1H), 4.18-4.16 (m, 2H), 4.09-4.07 (m, 2H), 3.87-3.85 (m, 3H), 3.71 (s, 3H), 3.56-3.52 (m, 1H), 3.45-3.43 (m, 1H), 3.38 (s, 3H), 3.11-3.08 (m, 3H), 2.60-2.58 (m, 2H), 2.53 (s, 2H), 1.29 (s, 6H), 1.17 (d, J = 6.0 Hz, 3H).
Example 304a 5-(2-Methoxyethyl)nitro-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazine 304a O Br O O N O N N N N Br N NH Pd/C, H2, MeOH N N O rt,2 h N NO2 N NH2 Pd2(dba)3, Xantphos, N Cs2CO3 Br dioxane, 100 oC, 3h 304a 304b 304c O O N N Pin2B2 N N N NH HN N 282i O O O Pd2(dba)3, N X-PHos, KOAc N N HO N Pd(dppf)Cl2, S dioxane, B K3PO4, NaOAc O N 70 oC, 2 h HO ACN, water, 100 oC, 1 h 10 304d 304e To a solution of 2-nitro-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine (190 mg, 1.13 mmol) 125i in itrile (10 mL) was added K2CO3 (311.9 mg, 2.26 mmol) and 1-bromo methoxyethane (188.3 mg, 1.36 mmol). The reaction mixture was heated at 80oC for 17 h under microwave irradiation. Analysis of reaction mixture by LCMS showed complete 15 conversion to the d product. The mixture was cooled to room temperature and ed.
The filtrate was concentrated under reduced pressure to afford 304a as a white solid (230 mg, 90%), which was used in the next step without further purification. MS-ESI: [M+H]+ 227.0 e 304b 5-(2-Methoxyethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin- 2-amine 304b 433 To a solution of 304a (286 mg, 1.26 mmol) in methanol (10 mL) was added Pd/C (28.6 mg). The system was evacuated and then ed with H2. After stirring at room temperature for 2 h, the mixture was filtered off. The filtrate was concentrated under reduced re to afford 304b as a yellow solid (240 mg, 97%), which was used in the next step 5 without further cation. MS-ESI: [M+H]+ 197.0 Example 304c 5-Bromo(5-(2-methoxyethyl)-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)methylpyridin-2(1H)-one 304c A 100-mL round-bottomed flask equipped with a magnetic r and a reflux condenser was charged with 304b (230 mg, 1.17 mmol), 3,5-dibromomethylpyridin- 10 one (468.4 mg, 1.76 mmol), Pd2(dba)3 (53.5 mg, 0.0585 mmol), Xantphos (67.6 mg, 0.117 mmol), Cs2CO3 (762.8 mg, 2.34 mmol), and dioxane (20 mL). After three cycles of vacuum/N2 flush, the mixture was heated at 100oC for 3 h. Analysis of the reaction mixture by LCMS showed complete conversion to the desired product. It was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The e 15 was purified by silica-gel column chromatography eluting with 40:1 romethane/methanol to afford 304c as a dark solid (380 mg, 85%). MS-ESI: [M+H]+ 382.2 Example 304d 3-(5-(2-Methoxyethyl)-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinylamino)methyl(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin- 20 2(1H)-one 304d A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 304c (330 mg, 0.86 mmol), Pin2B2 (329mg, 1.30 mmol), Pd2(dba)3 (40 mg, 0.043 mmol), X-phos (41 mg, 0.086 mmol), ium acetate (169 mg, 1.726 mmol), and dioxane (10 mL). After three cycles of /N2 flush, the mixture was 25 heated at 70oC for 2 h. Analysis of reaction mixture by LCMS showed complete conversion to the desired product. It was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue was washed with petroleum ether to afford 304d as a dark oil (240 mg, 80%), which was used in the next step without further purification. MS-ESI: [M+H]+ 348.3 30 Example 304e 2-{4,4-Dimethyloxothia-10,11- diazatricyclo[6.4.0.02,6]dodeca-1(8),2(6),11-trienyl}(5-{[5-(2-methoxyethyl)- 4H,5H,6H,7H-pyrazolo[1,5-a]pyrazinyl]amino}methyloxo-1,6-dihydropyridin yl)pyridinecarbaldehyde 304e 434 A 50-mL round-bottomed flask equipped with a reflux condenser was charged with 4- chloro{4,4-dimethyloxothia-10,11-diazatricyclo[6.4.0.02,6]dodeca-1(8),2(6),11- trienyl}pyridinecarbaldehyde 282i (60 mg, 0.167 mmol), 304d (143.4 mg, 0.334 mmol), Pd(dppf)Cl2 (6.8 mg, 0.0084 mol), K3PO4 (70.8 mg, 0.334 mmol), sodium acetate 5 (27.4 mg, 0.334 mmol), acetonitrile (10 mL), and water (3 drops). The system was subjected to three cycles of vacuum/nitrogen flush and heated at 100oC for 1 h under N2 protection.
Analysis of reaction mixture by LCMS showed complete conversion to the desired product. It was cooled to room temperature and filtered. The filtrate was trated under reduced pressure. The residue was purified by -gel column chromatography g with 40:1 10 dichloromethane/methanol to afford 304e (80 mg, 77%) as white solid. MS-ESI: [M+H]+ 626.8 Example 304 3-[3-(hydroxymethyl)[5-[[5-(2-methoxyethyl)-6,7-dihydro-4H- pyrazolo[1,5-a]pyrazinyl]amino]methyloxopyridyl]pyridyl]-7,7-dimethyl-6,8- dihydrocyclopenta[3,4]thieno[1,3-d]pyridazinone 304 15 To a solution of 304e (80 mg, 0.128 mmol) in dichloromethane (4 mL) and methanol (4 mL) was added NaBH4 (9.7 mg, 0.256 mmol). The reaction mixture was stirred at room temperature for 1 h. It was quenched with aqueous NH4Cl (10 mL) and concentrated under reduced pressure and the residue was extracted with dichloromethane (3 X 20 mL). The combined extract was dried over Na2SO4 and concentrated under reduced pressure. The 20 residue was ed by reverse-phase prep-HPLC to afford 304 (23.5 mg, 29%) as white solid. MS-ESI: [M+H]+ 628.8. 1H NMR (500 MHz, DMSO-d6) δ 8.57 (d, J = 4.5 Hz, 1H), 8.46 (s, 1H), 8.23 (s, 1H), 8.05 (d, J = 2.0 Hz, 1H), 7.52 (d, J = 5.5 Hz, 1H), 7.40 (d, J = 2.5 Hz, 1H), 5.89 (s ,1H), 4.87 (s, 1H), 4.39 (s, 2H), 3.92-3.90 (m, 2H), 3.61 (s, 2H), 3.59 (s, 3H), 3.50 (t, J = 5.5 Hz, 2H), 3.26 (s, 3H), .89 (m, 4H), 2.81 (s, 2H), 2.68-2.66 (m, 2H), 25 1.29 (s, 3H), 1.28 (s, 3H).
Example 305a ro(5-(2-methoxyethyl)-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)methylpyridazin-3(2H)-one 305a 435 A 100-mL round-bottomed flask equipped with a reflux condenser was d with 5-(2-methoxyethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinamine 304b (392 mg, 2.0 mmol), 3,5-dibromomethylpyridin-2(1H)-one (446 mg, 2.0 mmol), cesium carbonate (1.30 5 g, 4.0 mmol), and 1,4-dioxane (40 mL). After bubbling nitrogen through the suspension for 10 minutes, xantphos (115 mg, 0.20 mmol) and tris(dibenzylideneacetone)dipalladium(0) (92 mg, 0.10 mmol) were added. The system was subjected to three cycles of vacuum/argon flush and heated at 100 oC for 5 h. It was then cooled to room temperature and filtered. The solid was washed with dichloromethane (2 x 15 mL). The combined filtrate was concentrated 10 under reduced pressure. The residue was ed by silica-gel column chromatography eluting with dichloromethane/methanol (80/1 to 30/1) to afford 305a (412 mg, 61%) as a yellow solid. MS-ESI: [M+H]+ 338.9 Example 305b (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}(5-{[5-(2-methoxyethyl)-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin 15 yl]amino}methyloxo-1,6-dihydropyridazinyl)pyridinyl)methyl Acetate 305b A 25-mL round-bottomed flask equipped with a reflux condenser was charged with 305a (200 mg, 0.60 mmol), {3-[(acetoxy)methyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (238 mg, 0.60 mmol), K3PO4 (254 mg, 1.2 mmol), sodium acetate (98 mg, 1.2 mmol) and Pd(dppf)Cl2 20 (22 mg, 0.030 mmol), and itrile/water (8/0.5 mL). The system was subjected to three cycles of vacuum/nitrogen flush and heated at 100oC under N2 protection for 1 h. is of the reaction mixture by LCMS showed complete conversion to the desired product. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue was partitioned between dichloromethane (30 mL) and 25 water (20 mL). The water layer was extracted with romethane (2 × 30 mL). The ed organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. The dark residue was purified by silica-gel column chromatography eluting with 436 dichloromethane/methanol (80/1 to 30/1) to afford 305b (169 mg, 43%) as a yellow solid.
MS-ESI: [M+H]+ 655.9 Example 305 3-[3-(hydroxymethyl)[5-[[5-(2-methoxyethyl)-6,7-dihydro-4H- lo[1,5-a]pyrazinyl]amino]methyloxo-pyridazinyl]pyridyl]-7,7-dimethyl- 5 1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 305 To a on of 305b (160 mg, 0.24 mmol) in THF/i-propanol /water(6/4/3mL) was added lithium hydroxide (29 mg, 1.2 mmol). The mixture was stirred at room temperature for 1 h and concentrated under reduced pressure. The residue was partitioned n water (15 mL) and ethyl acetate (20 mL). The water phase was extracted with ethyl acetate (3 X 20 mL). 10 The combined organic layer was dried and trated under pressure. The residue was purified by reverse-phase prep-HPLC to afford 305 as a white solid (88 mg, 60%). MS-ESI: [M+H]+ 614.3. 1H NMR (500 MHz, CDCl3) δ 8.56 (d, J = 5.0 Hz, 1H), 7.99 (s, 1H), 7.87 (s, 1H), 7.44 (d, J = 5.0 Hz, 1H), 6.84 (s, 1H), 5.96 (s, 1H), 4.59-4.57 (m, 3H), 4.49-4.47 (m, 1H), 4.17-4.12 (m, 4H), 3.92-3.90 (m, 4H, p), 3.77-3.75 (m, 2H), 3.61-3.60 (m, 2H), 15 3.41 (s, 3H), 3.05-3.03 (m, 2H), 2.81-2.79 (m, 2H), 2.60-2.58 (m, 2H), 2.52-2.50 (m, 2H), 1.28 (s, 6H). e 306a (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{5-[(6-methoxypyridazinyl)amino]methyloxo-1,6- dihydropyridinyl}pyridinyl)methyl Acetate 306a 20 A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with oxypyridazinamine (65 mg, 0.52 mmol), XantPhos (29 mg, 0.050 mmol), Pd2dba3 (45 mg, 0.050 mmol), [4-(5-bromomethyloxo- 1,6-dihydropyridinyl){4,4-dimethyloxo-1,10-diazatricy-clo[6.4.0.02,6]dodeca-2(6),7- 25 dienyl}pyridinyl]methyl acetate 273a (281 mg, 0.52 mmol), Cs2CO3 (326 mg, 1.0 mmol), and 1,4-dioxane (10 mL). After bubbling nitrogen through the resulting mixture for 20 minutes, it was heated at reflux for 2 h. After the completion of the reaction, the e was evaporated under reduced pressure and the residue was partitioned between ethyl acetate (20 mL) and water (10 mL). The ethyl acetate was concentrated under reduced pressure and 437 the residue was purified by -gel column chromatography eluting with 20:1 romethane/methanol to afford 306a (180 mg, 60%). MS-ESI: [M+H]+ 584.3.
Example 306 3-[3-(hydroxymethyl)[5-[(6-methoxypyridazinyl)amino] methyloxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- 5 tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 306 A mixture of 306a (180 mg, 0.32 mmol) and lithium hydroxide monohydrate (84 mg, 2.0 mmol) in THF (5 mL), i-propanol (5 mL), and water (1.5 mL) was stirred at 40ºC for 0.5 h. The mixture was evaporated under reduced pressure and the residue was partitioned between ethyl e (20 mL) and water (10 mL). The ethyl acetate was concentrated under 10 reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 306 (80 mg, 49%). : [M+H]+ 542.3. 1H NMR (500 MHz, DMSO-d6) δ 8.86 (s, 1H), 8.76 (d, J = 2.5 Hz, 1H), 8.51 (d, J = 5.0 Hz, 1H), 7.73 (d, J = 10.0 Hz, 1H), 7.56 (d, J = 2.5 Hz, 1H), 7.35 (d, J = 5.0 Hz, 1H), 7.11-7.10 (m, 1H), 6.56 (s, 1H), 4.94-4.93 (m, 1H), .45 (m, 1H), 4.40-4.38 (m, 1 H), 4.25-4.18 (m, 3H), 3.91 (s, 3H), 3.86-3.84 (m, 1H), 3.62 (s, 3H), 15 2.58-2.56 (m, 2H), 2.50-2.49 (m, 2H), 1.21 (s, 6H).
Example 307a 1,3-Dimethylnitro-1H-indazole 307a To a solution of 1-(2-chloronitrophenyl)ethanone (500 mg, 2.5 mmol) in anhydrous ethanol (15 mL) was added 1,1-dimethylhydrazine hydrochloride (3.38 g, 35.0 20 mmol) under nitrogen protection. The mixture was heated at reflux for 10 h and evaporated 438 under reduced re to afford crude 307a (3.0 g), which was used in the next step without further purification. MS-ESI: [M+H]+ 192.2 Example 307b 1,3-Dimethyl-1H-indazolamine 307b To a solution of 307a (crude, 2.5 mmol ) in ethanol (95%, 30 mL) was added 5 NH2NH2.water (1.25 g, 25.0 mmol), Pd/C (100 mg) under nitrogen protection. The mixture was stirred at 50 oC for 1.5 h. It was then cooled to room temperature and filtered through a pad of CELITE®. The filtrate was concentrated under d pressure and the residue was recrystallized from anhydrous ethanol (5 mL) to afford 307b as white solid (340 mg, 84% over two steps). MS-ESI: [M+H]+ 162.3 10 Example 307c o(1,3-dimethyl-1H-indazolylamino methylpyrazin-2(1H)-one 307c To a solution of 307b (280 mg, 1.74 mmol) in i-propanol (7 mL) was added triethylamine (352 mg, 3.48 mmol) and 3,5-dibromomethylpyrazin-2(1H)-one (H-005) (700 mg, 2.61 mmol). After being stirred at reflux for 6 h, the e was cooled to room 15 temperature. The precipitate was filtered, washed with anol (2 X 2 mL), and dried at 60oC under reduced pressure to afford 307c as a brown solid (560 mg, 92%). MS-ESI: [M+H]+ 347.8.
Example 307d (4-{6-[(1,3-Dimethyl-1H-indazolyl)amino]methyloxo- 4,5-dihydro-pyrazinyl}{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 20 2(6),7-dienyl}pyridinyl)methyl Acetate 307d A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 307c (300 mg, 0.86 mmol), 1,4-dioxane (20 mL), water (1 mL), [4-(dihydroxyboranyl){4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}pyridinyl]methyl acetate 199e (512 mg, 1.28 mmol), and cesium 25 carbonate (560 mg, 1.72 mmol). After bubbling nitrogen through the sion for 10 minutes, Cy3P (96 mg, 0.34 mmol) and Pd2(dba)3 (79 mg, 0.086 mmol) were added. The system was ted to three cycles of vacuum/argon flush and heated at reflux for 3 h. It was then cooled to room temperature and filtered. The solid was washed with dichloromethane (3 X 20 mL). The combined filtrate was trated under reduced 30 pressure and the residue was purified by silica-gel column chromatography eluting with 100:1 dichloromethane/methanol to afford 307d (230 mg, 43%) as a yellow solid. MS-ESI: [M+H]+ 620.9 439 Example 307 3-[4-[6-[(1,3-dimethylindazolyl)amino]methyloxo-pyrazin yl](hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5- b]pyrazinone 307 A mixture of 307d (230 mg, 0.37 mmol), m hydroxide (89 mg, 3.7 mmol) in i- 5 propanol /THF (1:1, 10 mL) and water (2 mL) was d at room temperature for 1 h. It was filtered and the filtrate was concentrated under reduced re. The resulting e was purified by reverse-phase prep-HPLC to afford 307 (41 mg, 19%) as a white solid. MS-ESI: [M+H]+ 578.4. 1H NMR (500 MHz, CDCl3) δ 8.58 (d, J = 2.0 Hz, 1H), 8.56 (d, J = 5.0 Hz, 1H), 8.37 (s, 1H), 8.11 (s, 1H), 7.88 (d, J = 5.0 Hz, 1H), 7.51 (dd, J = 2.0, 9.0 Hz, 1H), 7.33 10 (d, J = 9.0 Hz, 1H), 6.86 (s, 1H), .14 (m, 1H), 4.75-4.73 (m, 1H), 4.55-4.52 (m, 1H), 4.48-4.43 (m, 1H), 4.20-4.16 (m, 2H), 4.02 (s, 3H), 3.92-3.70 (m, 1H), 3.70 (s, 3H), 2.60 (d, J = 7.0 Hz, 2H), 2.57 (s, 3H), 2.54 (s, 2H), 1.30 (s, 6H).
Example 308a 2-{4,4-Dimethyloxothiaazatricyclo[6.4.0.02,6]dodeca- 1(8),2(6)-dienyl}(5-{[5-(2-methoxyethyl)-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin 15 yl]amino}methyloxo-1,6-dihydropyridinyl)pyridinecarbaldehyde 308a O N N HN N O O S N N O N 308a Following the procedure in Example 304, and starting with 4-chloro{4,4- dimethyloxothiaazatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienyl}pyridine carbaldehyde 109a (250 mg, 0.693 mmol) and 3-(5-(2-methoxyethyl)-4,5,6,7- 20 tetrahydropyrazolo[1,5-a]pyrazinylamino)methyl(4,4,5,5-tetramethyl-1,3,2- dioxaborolanyl)pyridin-2(1H)-one 304d (595 mg, 0.1.386 mmol), 308a was obtained as a yellow solid (250 mg, 57%). MS-ESI: [M+H]+ 628.3 Example 308 3-[3-(hydroxymethyl)[5-[[5-(2-methoxyethyl)-6,7-dihydro-4H- pyrazolo[1,5-a]pyrazinyl]amino]methyloxopyridyl]pyridyl]-7,7-dimethyl- 25 1,2,6,8-tetrahydrocyclopenta[3,4]thieno[1,3-c]pyridinone 308 Following the procedures in Example 304, and starting with 308a (230 mg, 0.366 mmol) and NaBH4 (27.7 mg, 0.732 mmol ), 308 was obtained as a white solid (53.2 mg, 440 23%). MS-ESI: [M+H]+ 629.8. 1H NMR (500 MHz, DMSO-d6) δ 8.49 (d, J = 5.0 Hz, 1H), 8.19 (s, 1H), 8.04 (d, J = 2.0 Hz, 1H), 7.37 (d, J = 2.5 Hz, 1H), 7.34 (d, J = 5.0 Hz, 1H), 5.89 (s, 1H), 4.95-4.93 (m, 1H), 4.47-4.39 (m, 2H), 4.20-4.14 (m, 1H), 3.92-3.91 (m, 2H), 3.84- 3.80 (m, 1H), 3.61 (s, 2H), 3.58 (s, 3H), 3.51-3.49 (m, 2H), 3.25 (s, 3H), .00 (m, 1H), 5 2.91-2.87 (m, 3H), 2.77 (s, 2H), 2.68-2.53 (m, 4H), 1.19 (s, 3H), 1.18 (s, 3H).
Example 309a 5-(3-Methoxypropyl)nitro-4,5,6,7-tetrahydropyrazolo[1,5- zine 309a A ave vial equipped with a magnetic stirrer was charged with 1-(2- 10 bromoethyl)(chloromethyl)nitro-1H-pyrazole 296c (600 mg, 2.2 mmol), 3- methoxypropanamine (595 mg, 6.6 mmol), and DMSO (6 mL). It was heated at 120 ºC under microwave irradiation for 0.5 h. The mixture was then cooled to room temperature and diluted with ethyl acetate (30 mL). The resulting mixture was washed with water (3 X 10 ml).
The organic layer was dried and filtered. The filtrate was concentrated under pressure and the 15 residue was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 309a (350 mg, 66%) as a yellow solid. MS-ESI: [M+H]+ 241.1 e 309b 5-(3-Methoxypropyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin- 2-amine 309b 441 A solution of 309a (300 mg, 1.25 mmol) in ethanol (20 mL) was added Pd/C (10%, 30 mg). The reaction was charged with hydrogen gas (via balloon) and stirred at room temperature for 1 h. After the reaction was complete, the mixture was filtered through a plug of CELITE®. The filtrate was trated under reduced pressure to afford 309b as a 5 yellow solid (250 mg, 92%), which was used without further purification in the next step.
MS-ESI: [M+H]+ 211.3 Example 309c 5-Bromo(5-(3-methoxypropyl)-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinyl-amino)methylpyridin-2(1H)-one 309c A 100-mL single-neck round-bottomed flask equipped with a magnetic r and a 10 reflux condenser was charged with 3,5-dibromomethylpyridin-2(1H)-one (320 mg, 1.2 mmol), 309b (250 mg, 1.2 mmol), tris(dibenzylideneacetone) dipalladium(0) (55 mg, 0.060 mmol), xantphos (70 mg, 0.12 mmol), cesium carbonate (782 mg, 2.4 mmol), and 1,4- dioxane (20 mL). The system was subjected to three cycles of vacuum/argon flush and heated at 100ºC for 5 h. It was then cooled to room temperature and filtered. The solid was washed 15 with dichloromethane (2 x 10 mL). The combined filtrate was concentrated under reduced pressure. The residue was ed by silica-gel column chromatography eluting with dichloromethane/methanol (80:1 to 30:1) to afford 309c (200 mg, 42%) as yellow solid. MSESI : [M+H]+ 396.2 Example 309d 4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 20 2(6),7-dien yl}(5-{[5-(3-methoxypropyl)-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin yl]amino}methyloxo-1,6-dihydropyridinyl)pyridinyl)methyl e 309d A 25-mL round-bottomed flask equipped with a reflux condenser was charged with 309c (120 mg, 0.30 mmol), cetoxy)methyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (240 mg, 25 0.60 mmol), K3PO4 (127 mg, 0.60 mmol), sodium acetate monohydrate (82 mg, 0.60 mmol), Pd(dppf)Cl2 (12 mg, 0.015 mmol), and acetonitrile/water (8/0.5 mL). The system was subjected to three cycles of vacuum/nitrogen flush and heated at 100oC under N2 tion for 2 h. Analysis of the reaction mixture by LCMS showed complete conversion to the desired t. The reaction mixture was cooled to room temperature and concentrated 30 under reduced pressure. The residue was partitioned between dichloromethane (20 mL) and water (10 mL). The water layer was extracted with romethane (2 × 10 mL). The combined organic t was dried over Na2SO4, filtered, and concentrated under reduced pressure. The dark residue was purified by silica-gel column chromatography eluting with 442 dichloromethane/methanol (80:1 to 30:1) to afford 309d (150 mg, 74%) as yellow solid. MSESI : [M+H]+ 668.9 Example 309 3-[3-(hydroxymethyl)[5-[[5-(3-methoxypropyl)-6,7-dihydro-4H- lo[1,5-a]pyrazinyl]amino]methyloxopyridyl]pyridyl]-7,7-dimethyl- 5 1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 309 To a solution of 309d (120 mg, 0.18 mmol) in THF/i-propanol (5/3/3mL) was added lithium hydroxide monohydrate (76 mg, 1.8 mmol). The mixture was stirred at 30 ºC for 1 h. After the reaction was complete, the mixture was filtered and the solvent was evaporated under reduced pressure. The residue was purified by reverse-phase prep-HPLC to 10 afford 309 as a white solid (85 mg, 76%). MS-ESI: [M+H]+ 627.3. 1H NMR (500 MHz, CDCl3) δ 8.48 (d, J = 5.0 Hz, 1H), 7.95 (d, J = 2.0 Hz, 1H), 7.72 (d, J = 2.0 Hz, 1H), 7.42 (s, 1H), 7.35 (d, J = 5.0 Hz, 1H), 6.86 (s, 1H), 5.71 (s, 1H), 5.05 (t, J = 6.0 Hz, 1H), 4.66-4.64 (m, 1H), 4.52-4.50 (m, 1H), 4.36-4.32 (m, 1H), 4.17-4.16 (m, 2H), 4.08-4.06 (m, 2H), 3.88- 3.85 (m, 1H), 3.71 (s, 3H), .64 (m, 2H), 3.48 (t, J = 6.0 Hz, 2H), 3.36 (s, 3H), 2.93 (t, J 15 = 6.0 Hz, 2H), .62 (m, 2H), 2.59-2.58 (m, 2H), 2.53 (s, 2H), 1.87-1.83 (m, 2H), 1.29 (s, 6H).
Example 310a (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{1-methyl[(5-methyl-1,2-thiazolyl)amino]oxo-1,6- dihydropyridinyl}pyridinyl)methyl Acetate 310a 20 A 25-mL sealed tube was charged with [4-(5-bromomethyloxo-1,6- dihydropyridinyl){4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dien- 10-yl}pyridinyl]methyl acetate 273a (150 mg, 0.28 mmol), 5-methylisothiazolamine hydrochloride (55 mg, 0.33 mmol), Cs2CO3 (183 mg, 0.56 mmol), Pd2(dba)3 (27 mg, 0.030 25 mmol), XantPhos (35 mg, 0.060 mmol), and e (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at 110ºC under microwave irradiation for 0.5 hour. It was the cooled to room temperature and evaporated under reduced pressure. The 443 residue was purified by silica-gel column eluting with 20:1 methylene chloride/methanol to afford 310a as a yellow solid (50 mg, 31%). MS-ESI: [M+H]+ 573.2. e 310 3-[3-(hydroxymethyl)[1-methyl[(5-methylisothiazol yl)amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- 5 tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 310 To a solution of 310a (50 mg, 0.090 mmol) in THF/ i-propanol /water (4 mL/4 mL/1 mL) was added lithium hydroxide (21 mg, 0.90 mmol). The reaction mixture was stirred at room temperature for 0.5 h and concentrated under reduced pressure. The residue was diluted with water (10 mL) and extracted with ethyl acetate (3 X 20 mL). The combined organic 10 layer was dried with Na2SO4 and concentrated under reduced pressure to afford a yellow solid, which was purified by reverse-phase prep-HPLC to afford 310 as a yellow solid (20 mg, 43%). MS-ESI: [M+H]+ 530.8. 1H NMR (500 MHz, CDCl3) δ 8.61 (d, J = 2.0 Hz, 1H), 8.49 (d, J = 5.5 Hz, 1H), 8.05 (s, 1H), 7.93 (d, J = 2.0 Hz, 1H), 7.37 (d, J = 5.0 Hz, 1H), 6.84 (s, 1H), 6.51 (s, 1H), .06 (m, 1H), 4.66-4.47 (m, 2H), .27 (m, 1H), 4.17-4.12 (m, 15 2H), .82 (m, 1H), 3.71 (s, 3H), 2.57 (d, J = 6.0 Hz, 2H), 2.52-2.50 (m, overlap, 5H), 1.27 (s, 6H).
Example 311a (4-{5-[(5-Cyclopropyl-1,2-oxazolyl)amino]methyloxo- 1,6-dihydro-pyridinyl}{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7- dienyl}pyridinyl)methyl e 311a 20 A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 5-cyclopropylisoxazolamine (80 mg, 0.65 mmol), XantPhos (29 mg, 0.050 mmol), Pd2dba3 (45 mg, 0.050 mmol), bromomethyloxo- 1,6-dihydropyridinyl){4,4-dimethyloxo-1,10-diazatricy-clo[6.4.0.02,6]dodeca-2(6),7- 25 dienyl}pyridinyl]methyl acetate 273a (350 mg, 0.65 mmol), Cs2CO3 (390 mg, 1.2 mmol), and 1,4-dioxane (10 mL). After bubbling nitrogen through the resulting mixture for 10 minutes, it was heated at reflux for 2 h. The mixture was then evaporated under reduced 444 pressure and the residue was partitioned between ethyl acetate (20 mL) and water (10 mL).
The organic layer was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 20:1 dichloromethane/methanol to afford 311a (120 mg, 32%) as a brown solid. MS-ESI: [M+H]+ 583.2. 5 Example 311 5-[(5-cyclopropylisoxazolyl)amino]methyloxo pyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 311 A mixture of 311a (120 mg, 0.20 mmol) and lithium hydroxide monohydrate (80 mg, 2.0 mmol) in THF (5 mL), i-propanol (5 mL) and water (1.5 mL) was stirred at 40 ºC for 0.5 10 h. The e was evaporated under reduced pressure and the residue was diluted with water (5 mL). It was then extracted with ethyl acetate (2 x 10 mL). The combined extract was trated under reduced pressure and the residue was purified by reverse-phase prep- HPLC to afford 311 (65 mg, 58%) as pale yellow solid. MS-ESI: [M+H]+ 541.3. 1H NMR (500 MHz, DMSO-d6) δ 8.92 (s, 1H), 8.48 (d, J = 5.0 Hz, 1H), 7.99 (d, J = 2.0 Hz, 1H), 7.55 15 (d, J = 2.0 Hz, 1H), 7.31 (d, J = 5.0 Hz, 1H), 6.56 (s, 1H), 6.19 (s, 1H), 4.92-4.90 (m, 1H), 4.45-4.44 (m, 1H), 4.40-4.39 (m, 1H), 4.24-4.18 (m, 3 H), 3.86-3.83 (m, 1H), 3.59 (s, 3H), 2.58-2.56 (m, 2H), 2.44-243 (m, 2H), .04 (m, 1H), 1.22 (s, 6H) .99 (m, 2H), 0.84-0.81 (m, 2H).
Example 312a 5-Bromomethyl(5-methyl(oxetanyl)-1H-pyrazol 20 ylamino)pyridin-2(1H)-one 312a A mixture of omethyl(5-methyl-1H-pyrazolylamino)pyridin-2(1H)- one 115a (200 mg, 0.71 mmol), 3-iodooxetane (647 mg, 3.53 mmol), Cs2CO3 (1150 mg, 3.53 mmol), and acetonitrile (5 mL) was heated at 80 0C in a sealed tube overnight. The mixture 25 was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica-gel column tography eluting with 20:1 dichloromethane/methanol to afford the 312a as a yellow solid (120 mg, 50%).
MS-ESI: [M+H]+ 339.1 445 Example 312b (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}(1-methyl{[5-methyl(oxetanyl)-1H-pyrazolyl]amino} oxo-1,6-dihydropyridinyl)pyridinyl)methyl e 312b A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 5 reflux condenser was charged with 312a (170 mg, 0.50 mmol), (2-{4,4-dimethyloxo-1,{3- [(acetyloxy)methyl]{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dien- 10-yl}pyridinyl}boronic acid 199e (200 mg, 0.50 mmol), CH3COONa (82 mg, 1.00 mmol), PdCl2(dppf) (41 mg, 0.050 mmol), K3PO4 (212 mg, 1.00 mmol), acetonitrile (10 mL), and water (0.5 mL). After ng nitrogen through the resulting mixture for 20 minutes, it 10 was heated at 100 oC under nitrogen atmosphere for 2 h. The mixture was cooled to room temperature and filtered. The filtrate was evaporated under reduced pressure and the residue was purified by silica-gel column tography eluting with 50:1 dichloromethane/methanol to afford 312b as white solid (172 mg, 56%). MS-ESI: [M+H]+ 612.4 15 Example 312 3-[3-(hydroxymethyl)[1-methyl[[5-methyl(oxetan yl)pyrazolyl]amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 312 A mixture of 312b (90 mg, 0.15 mmol) and lithium ide (14 mg, 0.60 mmol) in i-propanol/THF/water (6/4/2 mL) was stirred at room temperature for 0.5 h. The mixture was 20 concentrated under reduced pressure. The residue was partitioned between water (10 mL) and romethane (3 X 20 mL). The combined dichloromethane extract was concentrated under reduced pressure and the residue was purified by e-phase prep-HPLC to afford 312 as a white solid (54 mg, 64%). MS-ESI: [M+H]+ 569.9. 1H NMR (500 MHz, DMSO-d6) δ 8.47 (d, J = 5.0 Hz, 1H), 8.24 (d, J = 2.0 Hz, 1H), 8.23 (s, 1H), 7.48 (d, J = 1.5 Hz, 1H), 25 7.36 (d, J = 5.0 Hz, 1H), 6.55 (s, 1H), 5.95 (s, 1H), 5.46-5.42 (m, 1H), 4.99-4.91 (m, 3H), 4.81-4.78 (m, 2H), 4.52-4.41 (m, 2H), 4.24-4.18 (m, 3H), 3.87-3.84 (m, 1H), 3.60 (s, 3H), 2.61-2.56 (m, 2H), 2.43 (s, 2H), 2.15 (s, 3H), 1.22 (s, 3H), 1.19 (s, 3H).
Example 313a Ethyl 2-(5-(Hydroxymethyl)nitro-1H-pyrazolyl)acetate 313a 446 O O N N O OH N+ -O A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with acetonitrile (30 mL), (3-nitro-1H-pyrazolyl)methanol (1.43 g, 10.0 mmol), Cs2CO3 (490 mg, 1.5 mmol), and ethyl 2-bromoacetate (2.00 g, 12 mmol). The mixture was 5 stirred at 40oC for 5 h. It was then cooled to room ature and filtered. The filtrate was concentrated under reduced pressure and the e was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 313a (1.65 g, 72%) as a yellow solid. MS-ESI: [M+H]+ 229.9 Example 313b Ethyl 2-(5-(Chloromethyl)nitro-1H-pyrazolyl)acetate 10 313b O O Cl N N O N+ O- To a mixture of 313a (1.50 g, 6.55 mmol) in CHCl3 (60 mL) cooled at 0oC was slowly added SOCl2 (2.34 g, 19.6 mmol) while maintaining the internal temperature below 5oC. This reaction mixture was warmed to 50oC and stirred at this temperature for 3 h. It was then 15 cooled to 0oC and quenched with water. The organic layer was separated and evaporated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 30:1 romethane/methanol to afford 313b (1.1 g, 68%) as a yellow solid.
MS-ESI: [M+H]+ 247.9 e 313c 5-Methylnitro-4,5-dihydropyrazolo[1,5-a]pyrazin-6(7H)-one 20 313c 447 To a solution of 313b (1.0 g, 4.0 mmol) in dichloromethane (30 mL) was added a solution of CH3NH2 (1.07 g, 12.0 mmol, 35% in methanol). This reaction mixture was stirred at room temperature for 3 h and diluted with water (30 mL). The c layer was separated, dried over Na2SO4, and concentrated under reduced pressure. The residual was ed by 5 silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 313c (450 mg, 57%) as a yellow solid. MS-ESI: [M+H]+ 196.9 Example 313d 2-Aminomethyl-4,5-dihydropyrazolo[1,5-a]pyrazin-6(7H)- one 313d O N N N H2N 10 A solution of 313c (450 mg, 2.3 mmol) in ethanol (30 mL) was added Pd/C (10%, 400 mg). The reaction was charged with hydrogen gas (via balloon) and d at room temperature for 2 h. After reaction was complete, the mixture was filtered through a plug of CELITE® and the filtrate was concentrated under reduced pressure to afford 313d as a yellow solid (320 mg, 84%), which was used without further purification in the next step. 15 MS-ESI: [M+H]+ 167.1 Example 313e 2-(5-Bromomethyloxo-1,2-dihydropyridinylamino) methyl-4,5-dihydropyrazolo[1,5-a]pyrazin-6(7H)-one 313e A 100-mL round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 313d (300 mg, 1.8 mmol), 3,5-dibromomethylpyridin-2(1H)- 20 one (482 mg, 1.8 mmol), cesium ate (1.17 g, 3.6 mmol), and 1,4-dioxane (20 mL).
After bubbling nitrogen through the suspension for 10 minutes, xantphos (104 mg, 0.18 mmol) and tris(dibenzylideneacetone)dipalladium(0) (82 mg, 0.090 mmol) were added. The system was subjected to three cycles of vacuum/argon flush and heated at reflux for 5 h. It was then cooled to room temperature and filtered. The solid was washed with 25 dichloromethane (2 X 30 m). The combined filtrate was concentrated under d pressure.
The residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (80/1 to 30/1) to afford 313e (390 mg, 61%) as a yellow solid. e 313f (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}[1-methyl({5-methyloxo-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin 30 yl}amino)oxo-1,6-dihydropyridinyl]pyridinyl)methyl Acetate 313f 448 A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with 313e (150 mg, 0.43 mmol), {3-[(acetoxy)methyl]{4,4- dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic 5 acid 199e (170 mg, 0.43 mmol), K3PO4 (183 mg, 0.86 mmol), sodium acetate (71 mg, 0.86 mmol), Pd(dppf)Cl2 (35 mg, 0.043 mmol), acetonitrile (10 mL), and water (0.5 mL). The system was subjected to three cycles of vacuum/nitrogen flush and heated at 100oC for 3 h.
After this time the reaction was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column 10 chromatography eluting with 30:1 romethane/methanol to afford 313f (131 mg, 49%) as a yellow solid. MS-ESI: [M+H]+ 625.3.
Example 313 3-[3-(hydroxymethyl)[1-methyl[(5-methyloxo-4,7- dihydropyrazolo[1,5-a]pyrazinyl)amino]oxopyridyl]pyridyl]-7,7-dimethyl- 1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 313 15 A mixture of 313f (130 mg, 0.21 mmol) and lithium hydroxide (10 mg, 0.42 mmol) in i-propanol/THF (1:1, 7 mL) and water (2 mL) was stirred at 0oC for 0.5 h. The mixture was concentrated under reduced pressure. The residue was partitioned between water (10 mL) and ethyl acetate (3 X 10 mL). The combined ethyl e extract was trated under d pressure and the residue was purified with reverse-phase prep-HPLC to afford 313 20 (60 mg, 49%) as a white solid. MS-ESI: [M+H]+ 582.8. 1H NMR (500 MHz, DMSO-d6 ) δ 8.48 (d, J = 5.0 Hz, 1H), 8.37(s, 1H), 8.10 (d, J = 2.0 Hz, 1H), 7.43 (d, J = 2.5 Hz, 1H), 7.33 (d, J = 5.0 Hz, 1H), 6.57 (s, 1H), 6.07 (s, 1H), 4.95 (bs, 1H), 4.62-4.54 (m, 4H), 4.46-4.42 (m, 2H), 4.24-4.19 (m, 3H), 3.89-3.82 (m, 1H), 3.60 (s, 3H), 2.99 (s, 3H), 2.60-2.57 (m, 2H), 2.45-2.44 (m, 2H), 1.23 (s, 6H). 25 Example 314a 1-(6-Nitropyridinyl)azetidinol 314a A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with acetonitrile (50 mL), ronitropyridine (1.2 g, 7.9 mmol), K2CO3 (2.1 g, 15.8 mmol), and azetidinol hydrochloride(1.3 g, 11.9 mmol). The 449 mixture was heated at 60oC for 1 h. After this time the reaction was cooled to room temperature. It was then filtered and the filtrate was evaporated under reduced pressure. The residue was purified by -gel column chromatography eluting with dichloromethane/methanol (50:1 to 20:1) to afford 314a (1.1 g, 73%) as a yellow solid. MS- 5 ESI: [M+H]+196.0.
Example 314b minopyridinyl)azetidinol 314b A 100-mL single-neck round-bottomed flask was purged with nitrogen and charged with 314a (1.0 g, 5.1 mmol), 10% palladium on carbon (10% wet, 100 mg), and ethanol (40 mL). The e was evacuated, charged with hydrogen gas, and stirred at room temperature 10 for 5 h. The hydrogen was then evacuated and nitrogen was d into the flask. The catalyst was removed by filtration h a pad of CELITE® and the filtrate was concentrated under reduced pressure to afford 314b as a yellow solid (792 mg, 85%). MSESI : [M+H]+ 166.1.
Example 314c 5-Bromo(5-(3-hydroxyazetidinyl)pyridinylamino) 15 methylpyridin-2(1H)-one 314c A 100-mL single-neck round-bottomed flask ed with a magnetic stirrer and a reflux condenser was charged with 314b (792 mg, 4.8 mmol), 3,5-dibromomethylpyridin- 2(1H)-one (1.9 g, 7.2 mmol), tris-(dibenzylideneacetone)dipalladium(0) (440 mg, 0.48 mmol), XantPhos (555 mg, 0.96 mmol), Cs2CO3 (3.1 g, 9.6 mmol), and oxane (40 mL). After 20 three cycles of vacuum/argon flush, the mixture was heated at 90oC for 3.0 hrs. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (50:1 to 20:1) to afford 314c as a yellow solid (1.5 g, 89%). MSESI : [M+H]+ 351.1 25 Example 314d (2-{4,4-Dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}(5-{[5-(3-hydroxyazetidinyl)pyridinyl]amino}methyloxo- 1,6-dihydro pyridinyl)pyridinyl)methyl Acetate 314d 450 A 50-mL round-bottomed flask equipped with a reflux condenser was charged with 314c (176 mg, 0.50 mmol), {3-[(acetoxy) methyl]{4,4-dimethyloxo-1,10-diazatri- cyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl} boronic acid 199e (198 mg, 0.50 mmol), Pd(dppf)Cl2 (41 mg, 0.050 mmol), K3PO4 (212.0 mg, 1.0 mmol), sodium e 5 (82.0 mg, 1.0 mmol), water (0.5 mL), and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at 95oC for 1 hour. It was then cooled to room temperature and filtered. The filtrate was trated under reduced pressure and the ing residue was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford crude 314d as a brown solid, which was used in the next 10 step without further purification. MS-ESI: [M+H]+ 623.8.
Example 314 3-[4-[5-[[5-(3-hydroxyazetidinyl)pyridyl]amino]methyl oxopyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 314 A mixture of 314d (crude product 311.5 mg, 0.50 mmol) and m hydroxide 15 hydrate (300 mg, 12.5 mmol) in i-propanol/THF/ water (2:2:1,10 mL) was stirred at room temperature for 30 min. The mixture was trated under reduced pressure. The e was ioned between water (10 mL) and dichloromethane (3 X 10 mL). The combined extract was concentrated under reduced pressure and the residue was purified by reversephase prep-HPLC to afford 314 (46 mg, two step: 16%) as yellow solid. MS-ESI: [M+H]+ 20 581.9. 1H NMR (500 MHz, DMSO-d6) δ 8.54 (d, J = 2.5 Hz, 1H), 8.47 (d, J = 5.0 Hz, 1H), 7.78 (d, J = 2.5 Hz, 1H), 7.70 (s, 1H), 7.54 (d, J = 2.5 Hz, 1H), 7.35 (d, J = 5.0 Hz, 1H), 6.83- 6.81 (m, 2H), 6.75 (d, J = 8.5 Hz, 1H), .02 (m, 1H), 4.77-4.75 (m, 1H), 4.64-4.62 (m, 1H), 4.50-4.48 (m, 1H), 4.34-4.32 (m, 1H), 4.17-4.14 (m, 4H), 3.86-3.83 (m, 1H), 3.70 (s, 3H), 3.64-3.62 (m, 2H), 2.57-2.56 (m, 2H), 2.51 (s, 2H), 2.31-2.30 (m, 1H),1.27 (s, 6H). 25 Example 315 3-[3-(hydroxymethyl)[1-methyl[[1-methyl(pyrrolidine carbonyl)pyrazolyl]amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 315 Following the procedures of Example 273, and substituting (3-aminomethyl-1H- pyrazolyl)(pyrrolidinyl)methanone for 2-aminopyridine, 315 was prepared. 27.3 mg, 30 60% yield. 1H NMR (400 MHz, DMSO-d6) δ 8.48 (d, J = 5.1 Hz, 1H), 8.23 (s, 1H), 8.05 (d, J = 2.4 Hz, 1H), 7.43 (d, J = 2.4 Hz, 1H), 7.34 (d, J = 5.1 Hz, 1H), 6.55 (s, 1H), 6.46 (s, 1H), 4.95 (t, J = 5.2 Hz, 1H), 4.42 – 4.47 (m, 1H), 4.17 – 4.21 (m, 3H), 3.79 (s, 2H), 3.59 (s, 3H), 3.48 (dt, J = 11.1, 6.6 Hz, 3H), 3.27 (s, 2H), 2.57 (d, J = 7.5 Hz, 2H), 2.43 (s, 2H), 1.90 – 1.84 (m, 3H), 1.22 (s, 6H). ES-MS m/z 611.4 [M+1]. 451 Example 316 3-[3-(hydroxymethyl)[5-[[5-(methoxymethyl)methyl-pyrazol yl]amino]methyloxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 316 Following the procedures of Example 273, and tuting 5-(methoxymethyl) 5 methyl-1H-pyrazolamine for 2-aminopyridine, 316 was prepared. 43.2 mg, 84% yield. 1H NMR (400 MHz, 6) δ 8.47 (d, J = 5.0 Hz, 1H), 8.15 (s, 1H), 8.04 (d, J = 2.4 Hz, 1H), 7.39 (d, J = 2.1 Hz, 1H), 7.33 (d, J = 5.0 Hz, 1H), 6.55 (s, 1H), 6.11 (s, 1H), 4.96 – 4.90 (m, 1H), 4.38 – 4.46 (m, 1H), 4.38 (s, 2H), 4.19 (d, J = 9.8 Hz, 2H), 3.82 – 3.96 (m, 1H), 3.65 (s, 3H), 3.58 (s, 3H), 3.27 (s, 2H), 2.57 (d, J = 7.5 Hz, 2H), 2.43 (s, 2H), 1.22 (s, 6H). ES-MS 10 m/z 558.3 [M+1].
Example 317a 5-Methyl(1-methyloxo(4,4,5,5-tetramethyl-1,3,2- dioxaborolanyl)-1,2-dihydropyridinylamino)-4,5-dihydropyrazolo[1,5-a]pyrazin-6(7H)- one 317a 15 A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 2-(5-bromomethyloxo-1,2-dihydropyridin ylamino)methyl-4,5-dihydropyrazolo[1,5-a]pyrazin-6(7H)-one 313e (270 mg, 1.0 eq., 0.68 mmol), Pin2B2 (863.6 mg, 5.0 eq., 3.4 mmol), Pd2(dba)3 (62.4 mg, 0.1 eq., 0.068 mmol), XPhos (64.8 mg, 0.2 eq., 0.14 mmol), potassium acetate (200 mg, 3.0 eq., 2.04 mmol), and 20 e (15 mL). After three cycles of vacuum/argon flush, the mixture was heated at 65ºC for 3 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure to afford crude 317a, which was used in the next step without further purification. MS-ESI: [M+H]+ 399.9. e 317b 2-{4,4-Dimethyloxothiaazatricyclo[6.4.0.02,6]dodeca- 25 1(8),2(6)-dienyl}[1-methyl({5-methyloxo-4H,5H,6H,7H-pyrazolo[1,5- a]pyrazinyl}amino)oxo-1,6-dihydropyridinyl]pyridinecarbaldehyde 317b A 50-mL single-neck bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 317a (100 mg, 0.28 mmol), 4-chloro{4,4-dimethyl 452 oxothiaazatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienyl}pyridinecarbaldehyde 109a (112 mg, 0.28 mmol), Pd(dppf)Cl2 (22.9 mg, 0.028 mmol), K3PO4 (118.7 mg, 0.56 mmol), sodium acetate (45.9 mg, 0.56 mmol), water (0.5 mL), and acetonitrile (10 mL). After three cycles of vacuum/argon flush, the mixture was heated at reflux for 1 hour. It was then 5 cooled to room ature and filtered. The filtrate was concentrated under reduced pressure and the ing residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (50:1 to 30:1) to afford 317b as a yellow solid (60 mg, 36%). MSESI : [M+H]+ 597.8.
Example 317 3-[3-(hydroxymethyl)[1-methyl[(5-methyloxo-4,7- 10 opyrazolo[1,5-a]pyrazinyl)amino]oxopyridyl]pyridyl]-7,7-dimethyl- 1,2,6,8-tetrahydrocyclopenta[3,4]thieno[1,3-c]pyridinone 317 A mixture of 317b (60 mg, 0.10 mmol) and NaBH4 (11.3 mg, 0.30 mmol) in methanol (5 mL) was stirred at room ature for 30 min. The mixture was quenched with water (15 mL) and concentrated under reduced pressure. The residue was extracted with 15 dichloromethane (3 X 10 mL). The combined extract was concentrated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 317 (15 mg, 25%) as a yellow solid. MS-ESI: [M+H]+ 599.8. 1H NMR (500 MHz, CDCl3) δ 8.50 (d, J = 5.0 Hz, 1H), 8.0 (d, J = 2.0 Hz, 1H), 7.69 (d, J = 2.5 Hz, 1H), 7.50 (s, 1H), 7.34 (d, J = 5.0 Hz, 1H), 5.86 (s, 1H), 4.82-4.66 (m, 4H), 4.56 (s, 2H), 4.42-4.33 (m, 2H), 3.83-3.81 (m, 1H), 20 3.70 (s, 3H), 3.15 (s, 3H), 2.98-2.94 (m, 2H), 2.80 (s, 2H), 2.57-2.52 (m, 2H), 1.28 (s, 6H).
Example 318a {4-[5-({4,5-Dimethyl-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin no)methyloxo-1,6-dihydropyridinyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dien yl}pyridinyl}methyl Acetate 318a N N N N N NH LiOH N NH OAc O OH O N iPrOH/THF/H2O N N N N N O N O N 318a 318b 25 A 50-mL round-bottomed flask equipped with a reflux condenser was charged with methyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinamine 287i (123 mg, 1.0 eq., 0.74 mmol), [4-(5-bromomethyloxo-1,6-dihydropyridinyl){4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl]methyl acetate 273a (400 mg, 453 1.0 eq., 0.74 mmol), Pd2(dba)3 (68 mg, 0.1 eq., 0.074 mmol), Xantphos (86 mg, 0.2 eq., 0.148 mmol), Cs2CO3 (487 mg, 2.0 eq., 1.48 mmol), and dioxane (15 mL). After three cycles of vacuum/N2 flush, the mixture was stirred at 100oC for 2 hr. The mixture was cooled to room ature and filtered. The filtrate was concentrated under reduced pressure. The resulting 5 residue was purified by silica-gel column chromatography eluting with 20:1 ethyl acetate/methanol to afford 318a as a brown solid (221 mg, 48%). MS-ESI: [M+H]+ 624.9 Example 318b 10-{4-[5-({4,5-Dimethyl-4H,5H,6H,7H-pyrazolo[1,5- a]pyrazinyl}amino)methyloxo-1,6-dihydropyridinyl](hydroxymethyl)pyridin- 2-yl}-4,4-dimethyl-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienone 318b 10 A 25-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with 318a (200 mg, 1.0 eq., 0.32 mmol), lithium hydroxide (38 mg, 5.0 eq., 1.60 mmol), i-propanol /THF (8/8 mL), and water (2 mL). The e was stirred at room temperature for 1 h and concentrated under reduced pressure. The residue was added partitioned between water and dichloromethane. The combined organic layer was 15 concentrated under reduced pressure. The residue was purified by e-phase PLC to afford racemic mixture 318b as a yellow solid (91 mg, 43%).
Example 318 (R)(5-((4,5-dimethyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin yl)amino)-3'-(hydroxymethyl)methyloxo-1,6-dihydro-[3,4'-bipyridin]-2'-yl)-7,7- dimethyl-2,3,4,6,7,8-hexahydro-1H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazinone 318 20 Chiral HPLC (column: OZ-H, 100% methanol (0.1% ethyl acetate)) resolution of 318b separated enantiomers 318 and 319. 318: MS-ESI: [M+H]+ 582.8. 1H NMR (500 MHz, CDCl3) δ 8.49 (d, J = 5.0 Hz, 1H), 7.99 (s, 1H), 7.72 (d, J = 2.5 Hz, 1H), 7.44 (s, 1H), 7.36 (d, J = 5.0 Hz, 1H), 6.86 (s, 1H), 5.74 (s, 1H), 5.04 (t, J = 6.5 Hz, 1H), 4.66-4.64 (m, 1H), 4.52- 4.48 (m,1H), 4.36-4.34 (m,1H), .05 (m, overlap, 4H), 3.88-3.86 (m, 1H), 3.72 (s, 3H), 25 3.43-3.41 (m, 1H), 3.17-3.15 (m, 1H), 2.87-2.85 (m, 1H), .59 (m, 2H), 2.53 (s, 2H), 2.48 (s, 3H), 1.46 (d, J = 6.5 Hz, 3H), 1.29 (s, 6H). e 319 (S)(5-((4,5-dimethyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin yl)amino)-3'-(hydroxymethyl)methyloxo-1,6-dihydro-[3,4'-bipyridin]-2'-yl)-7,7- dimethyl-2,3,4,6,7,8-hexahydro-1H-cyclopenta[4,5]pyrrolo[1,2-a]pyrazinone 319 30 Chiral HPLC (column: OZ-H, 100% methanol (0.1% ethyl acetate)) resolution of c 318b separated enantiomers 318 and 319. 319: MS-ESI: [M+H]+ 582.8. 1H NMR (500 MHz, CDCl3) δ 8.49 (d, J = 5.0 Hz, 1H), 7.99 (s, 1H), 7.72 (d, J = 2.5 Hz, 1H), 7.44 (s, 1H), 7.36 (d, J = 5.0 Hz, 1H), 6.86 (s, 1H), 5.74 (s, 1H), 5.04 (t, J = 6.5 Hz, 1H), 4.67-4.65 (m, 1H), 4.52-4.48 (m,1H), 4.36-4.34 (m, 1H), 4.18-4.05 (m, overlap, 4H), 3.88-3.86 (m, 1H), 454 3.72 (s, 3H), 3.43-3.41 (m, 1H), 3.17-3.15 (m, 1H), 2.87-2.85 (m, 1H), 2.60-2.59 (m, 2H), 2.53 (s, 2H), 2.48 (s, 3H), 1.46 (d, J = 6.5 Hz, 3H), 1.30 (s, 6H).
Example 320a (6-Aminopyridinyl)((3R,5S)-3,5- dimethylmorpholino)methanone 320a Br O O O N N O Br N N NH O O N NH2 Pd2(dba)3 Xantphos, Cs2CO3 N dioxane, 100 oC Br 320a 320b O N O N NH AcO O N 199e N N Pd(dppf)Cl2, K3PO4 NaOAc, CH3CN/H2O O N 5 320c To a solution of (3S,5R)-3,5-dimethylmorpholine (1.15 g, 10 mmol) in DMF (15 mL) was added HATU (3.8 g, 10 mmol), DIPEA (2.6 g, 20 mmol), and 6-aminonicotinic acid (1.38 g, 10 mmol) at room temperature. The reaction mixture was stirred for 18 h. It was then filtered and the filtrate was purified with lash (A: 1‰NH4HCO3/water, B: 10 acetonitrile) to afford 320a ( 650 mg, 27% ) as a yellow solid. : [M+H]+ 236.1.
Example 320b 5-Bromo(5-((3R,5S)-3,5-dimethylmorpholine carbonyl)pyridinylamino)methylpyridin-2(1H)-one 320b A 50-mL round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 320a (160 mg, 1.0 eq., 0.68 mmol), 3,5-dibromo 15 methylpyridin-2(1H)-one (273 mg, 1.5 eq., 1.02 mmol), a)3 (64 mg, 0.1 eq., 0.070 mmol), Xantphos (79 mg, 0.2 eq., 0.14 mmol), Cs2CO3 (444 mg, 2.0 eq., 1.36 mmol), and dioxane (20 mL). After three cycles of vacuum/nitrogen flush, the mixture was heated at 100oC for 2 h. It was then cooled to room temperature and filtered. The filtrate was evaporated under reduced pressure and the resulting residue was ed by -gel column 20 chromatography eluting with ethyl acetate to afford 320b (190 mg, 66%) as a yellow solid.
MS-ESI: [M+H]+ 420.8. 455 Example 320c (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{5-[(5-{[(3R,5S)-3,5-dimethylmorpholinyl]carbonyl}pyridin yl)amino]methyloxo-1,6-dihydropyridinyl}pyridinyl)methyl Acetate 320c A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 5 reflux condenser was d with 320b (150 mg, 1.0 eq., 0.36 mmol), {3-[(acetoxy)methyl]- -dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridin yl}boronic acid 199e (286 mg, 2.0 eq., 0.72 mmol), PdCl2(dppf) (29 mg, 0.10 eq., 0.040 mmol), K3PO4 (153 mg, 2.0 eq., 0.72 mmol), sodium acetate (59 mg, 2.0 eq., 0.72 mmol), acetonitrile (10 mL), and water (0.2 mL). After three cycles of vacuum/nitrogen flush, the 10 mixture was heated at 90oC for 2 h. It was then cooled to room temperature and filtered. The filtrate was evaporated under reduced pressure and the ing residue was purified by silica-gel column chromatography eluting with 50:1 romethane/methanol to afford 320c (161 mg, 64%) as brown solid. MS-ESI: [M+H]+ 693.8 Example 320 3-[4-[5-[[5-[(3S,5R)-3,5-dimethylmorpholinecarbonyl] 15 pyridyl]amino]methyloxopyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl- 1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 320 A 25-mL single-neck round-bottomed flask ed with a magnetic stirrer was charged with 320b (145 mg, 1.0 eq., 0.21 mmol), lithium hydroxide (26 mg, 5.0 eq., 1.05 mmol), THF (4.0 mL), i-propanol (4.0 mL), and water (1.0 mL). The mixture was stirred at 20 room temperature for 1 h and filtered. The filtrate was concentrated under reduced pressure.
The resulting residue was diluted with water (10 mL) and extracted with dichloromethane (3 X 15 mL). The combined organic layer was trated under reduced pressure. The residue was ed by reverse-phase prep-HPLC to afford 320 (35 mg, 26%) as white solid. MSESI : [M+H]+ 651.9 25 Example 321a 6-Chloro(5-((3R,5S)-3,5-dimethylmorpholine carbonyl)pyridinylamino)methylpyridazin-3(2H)-one 321a 456 A 25-mL round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with (6-aminopyridinyl)((3R,5S)-3,5- dimethylmorpholino)methanone 320a (235 mg, 1.0 mmol), 4-bromochloro methylpyridazin-3(2H)-one (232 mg, 1.05 mmol), cesium carbonate (652 mg, 2.0 mmol), 5 and 1,4-dioxane (6.0 mL). After bubbling nitrogen through the suspension for 10 minutes, Xantphos (116 mg, 0.20 mmol) and tris(dibenzylideneacetone)dipalladium(0) (70 mg, 0.10 mmol) were added. The system was subjected to three cycles of vacuum/nitrogen flush and heated at reflux for 2.5 h. It was then cooled to room temperature and ed. The solid was washed with dichloromethane (3 X 10 ml). The combined c layer was concentrated 10 under reduced pressure. The residue was purified by silica-gel column chromatography eluting with petroleum ethyl acetate (2:1 to 1:10) to afford 321a (140 mg, 37%) as a yellow solid. MS-ESI: [M+H]+ 378.3 Example 321b (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{5-[(5-{[(3R,5S)-3,5-dimethylmorpholinyl]carbonyl}pyridin 15 yl)amino]methyloxo-1,6-dihydropyridazinyl}pyridinyl)methyl Acetate 321b A 50-mL round-bottomed flask equipped with a reflux condenser was charged with 321a (140 mg, 0.37 mmol), (2-{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridinyl)methyl acetate 199e (355 mg, 0.74 mmol), K3PO4 (157 mg, 0.74 mmol), sodium e (61 mg, 0.74 20 mmol), 1,1’-bis(diphenylphosphino) enedichloropalladium(II) (36 mg, 0.040 mmol), acetonitrile (10 mL), and water (0.2 mL). The system was subjected to three cycles of vacuum/nitrogen flush and heated at 100 °C under N2 protection for 1.5 h. Analysis of on mixture by LCMS showed complete conversion to the desired product. The reaction mixture was cooled to room temperature and concentrated under d pressure. The 25 residue was partitioned between dichloromethane (30 mL) and water (30 mL). The aqueous layer was separated and extracted with dichloromethane (3 × 20 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The dark residue was purified by gel-silica column chromatograpthe hy eluting with 60:1 dichloromethane/methanol to afford 320b (105 mg, 41%) as a black solid. : [M+H]+ 30 695.3 e 321 3-[4-[5-[[5-[(3S,5R)-3,5-dimethylmorpholinecarbonyl] pyridyl]amino]methyloxo-pyridazinyl](hydroxymethyl)pyridyl]-7,7-dimethyl- 1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 321 457 To a solution of 321b (105 mg, 0.15 mmol) in THF/i-propanol /water(2/1/0.5 mL) was added lithium ide (36 mg, 1.5 mmol) at room temperature. After the reaction was stirred for 3h, LCMS indicated the reaction was complete. Then the mixture was poured into water (25 mL) and extracted with dichloromethane (3 X 20 mL). The combined organic layer 5 was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under d pressure. The residue was purified by e-phase prep-HPLC (A: CO3/water, B: acetonitrile) to afford 321 (100 mg, 95%) as a white solid. MS-ESI: [M+H]+ 652.8. 1H NMR (500 MHz, DMSO-d6) δ 9.69 (s, 1H), 8.63 (s, 1H), 8.53 (d, J = 5.0 Hz, 1H), 8.31 (d, J = 1.5 Hz, 1H), 7.77-7.75 (m, 1H), 7.58 (d, J = 8.5 Hz, 1H), 7.41 (d, J = 5.0 Hz,1H), 6.56 (s, 10 1H), 4.78 (t, J = 5.5 Hz, 1H), 4.60-4.57 (m, 1H), 4.41-4.37 (m, 1H), 4.30-4.25 (m, 1H), 4.19 (d, J = 3.5 Hz, 2H), 4.01-4.00 (m, 2H), 3.92-3.88 (m, 1H), 3.82 (s, 3H), 3.65-3.61 (m, 2H), 3.56-3.53 (m, 2H), 2.61-2.58 (m , 2H), 2.42 (s, 2H), 1.25 (d, J = 6.0 Hz, 6H), 1.21 (s, 6H) Example 322a (5-((3R,5S)-3,5-Dimethylmorpholinecarbonyl)pyridin- 2-ylamino)methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9- 15 hexahydropyrazino[1,2-a]indol-2(1H)-yl)pyridinyl)methyl Acetate 322a A 25-mL round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with (6-aminopyridinyl)((3R,5S)-3,5- dimethylmorpholino)methanone 320a (120 mg, 0.50 mmol), (4-(5-bromomethyloxo- 20 1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indol-2(1H)- yl)pyridinyl)methyl acetate 217a (262 mg, 0.50 mmol), cesium carbonate (326 mg, 1.0 mmol), and oxane (6 mL). After bubbling nitrogen through the suspension for 10 s, Xantphos (58 mg, 0.10 mmol) and tris(dibenzylideneacetone)dipalladium(0) (45mg, 0.050 mmol) were added. The system was subjected to three cycles of vacuum/nitrogen flush 25 and heated at reflux for 2.5 h. It was then cooled to room temperature and filtered. The solid was washed with dichloromethane (3 X 10 mL). The combined filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography 458 eluting with dichloromethane/methanol (80/1 to 50/1) to afford 322a (200 mg, 59%) as a yellow solid. : [M+H]+ 680.3 Example 322 2-[4-[5-[[5-[(3S,5R)-3,5-dimethylmorpholinecarbonyl] pyridyl]amino]methyloxopyridyl](hydroxymethyl)pyridyl]-3,4,6,7,8,9- 5 hexahydropyrazino[1,2-a]indolone 322 To a solution of 322a (136 mg, 0.20 mmol) in THF/i-propanol /water(4/2/1 mL) was added lithium hydroxide (48 mg, 2.0 mmol) at room temperature. After the reaction was stirred for 2 h, LCMS indicated the reaction was complete. Then the mixture was poured into water (15 mL) and extracted with dichloromethane (3 X 15 mL). The combined c layer 10 was washed with brine (30 mL), dried over Na2SO4, ed, and concentrated under reduced pressure. The residue was purified by reverse-phase prep-HPLC (A: 1‰NH4HCO3/water, B: acetonitrile) to afford 322 (50 mg, 40%) as a white solid. MS-ESI: [M+H]+ 638.3. 1HNMR (500 MHz, DMSO-d6) δ 8.94 (s, 1H), 8.77 (d, J = 5.5 Hz,1H), 8.48 (d, J = 5.0 Hz, 1H), 8.20 (d, J = 2.0 Hz, 1H), 7.64-7.62 (m, 1H), 7.59 (d, J = 2.5 Hz, 1H), 7.37-7.35 (m, 2H), 6.57 (s, 15 1H), 4.95 (t, J = 4.5 Hz, 1H), 4.47-4.38 (m, 2H), .99 (m, 5H), 3.88-3.87 (m, 1H), 3.65- 3.61 (m, overlap, 5H), 3.56-3.53 (m, 2H), 2.66-2.56 (m, 2H), 2.47-2.44 (m, 2H), 1.80-1.79 (m, 2H), 1.70-1.66 (m, 2H), 1.25 (d, J = 6.0 Hz, 6H) Example 323a 5-(3-Methoxyazetidinyl)nitropyridine 323a Br O O N O Br N Pd/C, H2, MeOH N rt, 2h Pd2(dba)3, os, N NO2 N NH2 Cs2CO3, dioxane, 100 oC, 3h 323a 323b O O N N N NH 199e N NH OAc O N O Pd(dppf)Cl2, N N N K3PO4, NaOAc, Br ACN, H2O, O N 100 oC, 1 h 323c 323d 459 A 100-mL round bottomed flask was equipped with a reflux condenser was charged with 3-methoxyazetidine hydrochloride (1.0 g, 8.09 mmol), 5-bromonitropyridine (1.97 g, 9.71 mmol), Pd2(dba)3 (370.1 mg, 0.404 mmol), Xantphos (467.6 mg, 0.809 mmol), Cs2CO3 (7.9 g, 24.3 mmol), and dioxane (50 mL). After bubbling nitrogen through the reaction 5 mixture for 20 minutes, it was heated at 100oC under N2 protection for 3 h. Analysis of the reaction mixture by LCMS showed complete sion to the d t. It was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica-gel column chromatography eluting with 50:1 dichloromethane/methanol to afford 323a as a yellow solid (1.63 g, 96%). MS-ESI: [M+H]+ 10 210.2 Example 323b 5-(3-Methoxyazetidinyl)pyridinamine 323b To a solution of 323a (1.5 g, 7.17 mmol) in methanol (150 mL) was added 10 % Pd/C (150 mg). The system was evacuated and then refilled with H2. After stirring at room temperature for 2 h, the mixture was ed. The filtrate was trated under reduced 15 pressure to afford 323b as a yellow oil (1.2 g, 93%), which was used in next step t further purification. MS-ESI: [M+H]+ 180.1 Example 323c o(5-(3-methoxyazetidinyl)pyridinylamino) methylpyridin-2(1H)-one 323c A 100-mL round bottomed flask was equipped with a reflux condenser was charged 20 with 323b (1.2 g, 6.7 mmol), 3,5-dibromomethylpyridin-2(1H)-one (2.14 g, 8.04 mmol), Pd2(dba)3 (306.5 mg, 0.335 mmol), Xantphos (387.3 mg, 0.67 mmol), Cs2CO3 (4.37 g, 13.4 mmol), and dioxane (50 mL). After bubbling nitrogen through the reaction mixture for 20 minutes, it was heated at 100oC under N2 protection for 3 h. is of the reaction mixture by LCMS showed complete conversion to the d product. It was cooled to room 25 temperature and ed. The filtrate was concentrated under reduced pressure. The residue was washed with petroleum ether to afford 323c as a brown solid (1.16 g, 47%), which was used in next step without further purification. MS-ESI: [M+H]+ 364.8.
Example 323d (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}(5-{[5-(3-methoxyazetidinyl)pyridinyl]amino}methyloxo- 30 1,6-dihydropyridinyl)pyridinyl)methyl Acetate 323d A 50-mL round bottomed flask was equipped with a reflux condenser was charged with 323c (150 mg, 0.411 mmol), {3-[(acetoxy)methyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (326.5 mg, 0.822 mmol), Pd(dppf)Cl2 (16.8 mg, 0.0205 mmol), K3PO4 (174.3 mg, 0.822 mmol), sodium 460 acetate (67.5 mg, 0.822 mmol), acetonitrile (10 mL), and water (5 drops). After bubbling nitrogen through the reaction mixture for 20 minutes, it was heated 100oC under N2 protection for 1 h. Analysis of the reaction mixture by LCMS showed complete conversion to the desired product. It was cooled to room temperature and filtered. The filtrate was concentrated 5 under reduced pressure. The e was purified by silica-gel column chromatography eluting with 40:1 romethane/methanol to afford 323d as a yellow oil (180 mg, 68.7%).
MS-ESI: [M+H]+ 637.8 Example 323 3-[3-(hydroxymethyl)[5-[[5-(3-methoxyazetidinyl) pyridyl]amino]methyloxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- 10 tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 323 To a solution of 323d (160 mg, 0.251 mmol) in THF (5 mL), i-propanol (5 mL), and water (5 mL) was added lithium hydroxide (95 mg, 2.51 mmol). The reaction mixture was stirred at room temperature for 1 h. Analysis of the reaction e by LCMS showed complete conversion to the desired product. It was then concentrated under reduced re 15 and the residue was diluted with water (10 mL). The resulting mixture was extracted with romethane (3 x 15 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase prep- HPLC to afford 323 (33.8 mg, 23%) as a white solid. MS-ESI: [M+H]+ 595.8. 1H NMR (500 MHz, DMSO-d6) δ 8.55 (d, J = 2.0 Hz, 1H), 8.49 (d, J = 5.0 Hz, 1H), 8.31 (s, 1H), 7.48 (d, J 20 = 3.5 Hz, 1H), 7.45 (d, J = 2.0 Hz, 1H), 7.34 (d, J = 5.0 Hz, 1H), 7.21 (d, J = 8.5 Hz, 1H), 6.91 (dd, J = 2.5, 8.5 Hz, 1H), 6.56 (s, 1H), 4.96-4.94 (m, 1H), .39 (m, 2H), 4.31-4.19 (m, 4H), 4.03-4.00 (m, 2H), 3.85-3.83 (m, 1H), 3.60 (s, 3H), .53 (m, 2H), 3.23 (s, 3H), .54 (m, 2H), 2.44-2.42 (m, 2H), 1.22 (s, 6H).
Example 324a (6-Aminopyridinyl)((3S,5S)-3,5- 25 dimethylmorpholino)methanone 324a 461 O (S)(S) Br N O O O (S)(S) N Cl N N N NH O O Pd2(dba)3, xantphos N N NH2 Cs2CO3, e Cl N 100 °C 324a 324b O (S) N O (S) N NH AcO O N 199e N N N O N Pd(dppf)Cl2, K3PO4 NaOAc, CH3CN, 100 °C 324c To a solution of (3S,5S)-3,5-dimethylmorpholine (115 mg, 1.0 mmol) in DMF (2 mL) was added HATU (380 mg, 1.0 mmol), DIPEA (260 mg, 2.0 mmol), and 6-aminonicotinic acid (138 mg, 1.0 mmol) at room temperature. After stirring for 18 h, the reaction mixture 5 was ed and purified with lash (A: 1‰NH4HCO3/water, B: acetonitrile) to afford 324a (80 mg, 34% ) as a yellow solid. MS (ESI): 236.1 (M+H).
Example 324b 6-Chloro(5-((3S,5S)-3,5-dimethylmorpholine carbonyl)pyridineylamino)methylpyridazin-3(2H)-one 324b A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 10 reflux condenser was charged with oxane (8 mL), cesium carbonate (221 mg, 0.68 mmol), 324a (80 mg, 0.34 mmol), and 4-bromochloromethylpyridazin-3(2H)-one (80 mg, 0.36 mmol). After bubbling nitrogen through the sion for 5 minutes, Xantphos (40 mg, 0.068 mmol) and tris(dibenzylideneacetone)dipalladium(0) (24 mg, 0.034 mmol) were added. The system was subjected to three cycles of vacuum/nitrogen flush and heated at 15 reflux for 2.5 h. It was then cooled to room temperature and filtered. The solid was washed with dichloromethane (3 X 10 mL). The combined filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with petroleum ether/ethyl acetate (2/1 to 100% ethyl acetate) to afford 324b (40 mg, 31%) as a yellow solid. MS-ESI: [M+H]+ 378.3 462 Example 324c (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- -dienyl}{5-[(5-{[(3S,5S)-3,5-dimethylmorpholinyl]carbonyl}pyridin yl)amino]methyloxo-1,6-dihydropyridazinyl}pyridinyl)methyl e 324c A 50-mL round-bottomed flask equipped with a reflux condenser was charged with 5 324b (40 mg, 0.11 mmol), (2-{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridinyl)methyl acetate 199e (105 mg, 0.22 mmol), K3PO4 (47 mg, 0.22 mmol), sodium acetate (18 mg, 0.22 mmol), 1,1’-bis(diphenylphosphino)ferrocenedichloropalladium(II) (20 mg, 0.022 mmol), acetonitrile (10 mL), and water (6 drops). The system was subjected to three cycles of 10 vacuum/nitrogen flush and heated at 100 °C under N2 protection for 1.5 h. LCMS analysis showed complete conversion to the desired product. The on mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue was diluted with dichloromethane (50 mL) and water (50 mL). The s layer was separated and extracted with dichloromethane (3 × 20 mL). The ed extract was dried 15 over Na2SO4, filtered, and concentrated under reduced pressure. The dark residue was purified by silica-gel column tography eluting with 60:1 dichloromethane/methanol to afford 324c (40 mg, 52%) as a black solid. MS-ESI: [M+H]+ 695.3 Example 324 3-[4-[5-[[5-[(3S,5S)-3,5-dimethylmorpholinecarbonyl] pyridyl]amino]methyloxo-pyridazinyl](hydroxymethyl)pyridyl]-7,7-dimethyl- 20 1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 324 To a solution of 324c (40 mg, 0.057 mmol) in THF/ i-propanol /water(1/1/0.5 ml) was added lithium hydroxide (14 mg, 0.57 mmol) at room temperature. After the reaction was stirred for 3 h, LCMS indicated the reaction was complete. Then the mixture was poured into water (20 mL) and extracted with dichloromethane (3 X 15 mL). The combined c layer 25 was washed with brine (30 mL), dried over Na2SO4, ed, and concentrated under reduced pressure. The residue solid was purified by reverse-phase prep-HPLC (A: 1‰NH4HCO3/water, B: acetonitrile) to afford 324 (10 mg, 27.7%) as a white solid. MS-ESI: [M+H]+ 653.3. 1H NMR (500 MHz, DMSO-d6) δ 9.69 (s, 1H), 8.63 (s, 1H), 8.53 (d, J = 5.0, 1H), 8.31 (d, J = 1.5 Hz, 1H), 7.77-7.75 (m, 1H), 7.58 (d, J = 8.5 Hz, 1H), 7.41 (d, J = 5.0 Hz, 30 1H), 6.56 (s, 1H), 4.78 (t, J = 5.5 Hz, 1H), .57 (m, 1H), 4.41-4.37 (m, 1H), 4.30-4.25 (m, 1H), 4.19 (d, J = 3.5 Hz, 2H), 4.01 (s, 2H), 3.92-3.88 (m, 1H), 3.82 (s, 3H), 3.65-3.61 (m, 2H), 3.56-3.53 (m, 2H), 2.57 (d, J = 6.5 Hz, 2H), 2.42 (s, 2H), 1.25 (d, J = 6.0 Hz, 6H), 1.21 (s, 6H) Example 325a 3-Aminobromomethylpyridin-2(1H)-one 325a 463 85% NH2-NH2.H2O IBX, EA ethylene glycol Br reflux, O/N Br OH Br 165°C, 3.5 h O N N N N F N F H 325b 325c 325d N O NH2 MeI, K2CO3 Br Br Br MeCN, 30°C, 1h N N 325a N N + N N Pd2(dba)3, BINAP, , dioxane, 100°C, 2 h, sealed 325e 326a N N N N N NH N NH 199e AcO O O N N N N Br PdCl2(dppf), K3PO4 NaOAc, MeCN, O N 100 °C,1 h, H2O 326f 326g To a solution of 5-bromo(diphenylmethyleneamino)methylpyridin-2(1H)-one (3.82 g, 10.4 mmol) in ethyl acetate (10 mL) was added 4 M HCl/dioxane (7.8 mL, 31.3 mmol). The reaction mixture was stirred for 0.5 h and concentrated under reduced pressure. 5 The residue was washed with tert-butyl methyl ether and filtered. The solid was dissolved in ethyl acetate (10 mL) and water (10 mL). The pH of the resulting mixture was adjusted to between 7 and 8 by adding K2CO3 gradually. The water phase was ted and extracted with dichloromethane for three times. The combined organic layer was concentrated under reduced pressure to afford 325a as a yellow solid (1.1 g, 52%). MS-ESI: [M+H]+ 202.9. 10 Example 325b romofluoropyridinyl)ethanol 325b To a 250-mL 3-neck flask was added a THF solution (20 mL) of 2-bromo fluoropyridine (8.80 g, 50 mmol). At -78 oC, to the solution was added LDA (25.0 mL, 50 mmol, 2.5 M in THF) dropwise. After ng for 5 min, diisopropylamine (7.0 mL, 50 mmol) was added dropwise via a syringe and the mixture was stirred at -78 oC for 4 h. A THF 464 solution of acetaldehyde (11 mL, 55 mmol, 5M in THF) was added dropwise via a syringe.
The contents were d from the cold bath and stirred with warming to room temperature overnight. The mixture was diluted with water (150 mL) and vigorously stirred for 5 min.
The ts were concentrated under reduced re and the residue was extracted with 5 ethyl ether (3 x 150 mL). The combined c layer was dried over MgSO4, filtered, and concentrated under reduced pressure to afford yellow oil, which was purified by silica-gel column chromatography eluting with petroleum ether/ethyl acetate (10:1 to 5:1) to afford 325b (8.0 g, 72.7%) as a yellow solid. MS-ESI: [M+H]+ 220.1. 1H NMR (500 MHz, CDCl3) δ 8.15 (d, J = 1.5 Hz, 1H), 7.68 (d, J = 5.5 Hz, 1H), 5.17 (d, J = 6.5 Hz, 1H), 2.18-2.16 (m, 10 1H), 1.52 (d, J = 6.5 Hz, 3H).
Example 325c romofluoropyridinyl)ethanone 325c A mixture of 325b (7.5 g, 34.2 mmol) and 2-iodoxybenzoic acid (38.4 g, 137 mmol) in ethyl acetate (200 mL) was stirred at 85oC for 20 hrs. The reaction mixture was filtered and the te was concentrated under reduced pressure. The resulting residue was purified by 15 silica-gel column chromatography eluting with petroleum ether/ethyl e (20:1 to 10:1) to afford 325c (6.8 g, 92%) as a yellow oil. MS-ESI: [M+H]+ 217.9.
Example 325d 5-Bromomethyl-1H-pyrazolo[3,4-c]pyridine 325d To a 250-mL round-bottomed flask equipped with a reflux condenser was added dry ethylene glycol (30 mL) and 325c (4.3 g, 20 mmol). Then hydrazine hydrate (5.0 mL, 4.8 g, 20 81.6 mmol) was added se via a syringe. The mixture was heated at 165 oC for 3.5 h.
The resulting orange-tan mixture was cooled to room temperature and the contents were poured onto a stirring mixture of 100 mL ice/water (1:1), whereupon precipitation occurred.
After ng for 10 min, the off-white precipitate was collected, which was dried in vacuo to afford 325d as an off-white solid (3.1 g, 74%). MS-ESI: [M+H]+ 211.9. 25 Example 325e 5-Bromo-1,3-dimethyl-1H-pyrazolo[3,4-c]pyridine 325e and 5- Bromo-2,3-dimethyl-2H-pyrazolo[3,4-c]pyridine 326a A mixture of 325d (3.0 g, 14.2 mmol), CH3I (2.40 g, 17.0 mmol), and K2CO3 (2.9 g, 21.3 mmol) in acetonitrile (60 mL) was d at 30oC for 1 h. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The resulting residue was 30 purified by silica-gel column chromatography eluting with 8:1 petroleum ether/ethyl acetate to afford 325e (920 mg, 29.0%) as a white solid, and g with 2:1 petroleum ether/ethyl acetate to afford 326a (390 mg, 12.0%) as a gray solid. MS-ESI: [M+H]+ 226.1.
Example 325f 5-Bromo(1,3-dimethyl-1H-pyrazolo[3,4-c]pyridin ylamino)methylpyridin-2(1H)-one 325f 465 A sealed tube was d with 325e (202 mg, 1.0 mmol), 325a (337.5 mg, 1.5 mmol), Pd2(dba)3 (91.7 mg, 0.10 mmol), BINAP (124.6 mg, 0.20 mmol), cesium carbonate (650 mg, 2.0 mmol), and 1,4-dioxane (10 mL). After three cycles of vacuum/nitrogen flush, the sealed tube was heated at 100 oC for 2 hrs. It was then cooled to room temperature and 5 filtered. The filtrate was concentrated under reduced pressure. The resulting residue was purified by silica-gel column chromatography eluting with petroleum ether/ethyl acetate (5:1 to 2:1) to afford 325f (140 mg, 40%) as a yellow solid. : [M+H]+ 348.2.
Example 325g {4-[5-({1,3-Dimethyl-1H-pyrazolo[3,4-c]pyridin no)- 1-methyloxo-1,6-dihydropyridinyl]{4,4-dimethyloxo-1,10- 10 diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dien yl}pyridinyl}methyl Acetate 325g A 100-mL round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 325f (120 mg, 0.35 mmol), {3-[(acetoxy)methyl]{4,4- dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (417 mg, 1.05 mmol), Pd(dppf)Cl2 (29 mg, 0.035 mmol), K3PO4 (148.0 mg, 0.70 15 mmol), sodium acetate (57.4 mg, 0.70 mmol), water (0.5 mL), and itrile (15 mL). After three cycles of vacuum/nitrogen flush, the mixture was heated at reflux for 1 hr. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 325g as a yellow solid (70 mg, 33%). MS-ESI: [M+H]+ 20 620.8.
Example 325 3-[4-[5-[(1,3-dimethylpyrazolo[3,4-c]pyridinyl)amino]methyl oxopyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 325 A e of 325g (60 mg, 0.10 mmol) and lithium hydroxide (60 mg, 2.5 mmol) in i- 25 propanol /THF (1:1, 4 mL) and water (1 mL) was stirred at 35oC for 30 min. To the reaction mixture was added water (10 mL) and the ing mixture was concentrated under reduced pressure. The residue was extracted with dichloromethane three times. The combined organic layer was concentrated under reduced pressure and the resulting residue was purified by e-phase prep-HPLC to afford 325 as a yellow solid (20 mg, 31%). MS-ESI: [M+H]+ 30 578.8. 1H NMR (500 MHz, DMSO) δ 8.80 (s, 1H), 8.49 (d, J = 5.0 Hz, 1H), 8.36 (d, J = 2.0 Hz, 1H), 8.29 (s, 1H), 7.55 (s, 1H), 7.49 (d, J = 2.0 Hz, 1H), 7.37 (d, J = 5.0 Hz, 1H), 6.56 (s, 1H), 5.06-5.05 (m, 1H), 4.51-4.43 (m, 2H), .19 (m, 3H), 4.00 (s, 3H), 3.86-3.84 (m, 1H), 3.63 (s, 3H), 2.62-2.59 (m, 2H), 2.44-2.43 (m, overlap, 5H), 1.22 (s, 6H). 466 Example 326b 5-Bromo(2,3-dimethyl-2H-pyrazolo[3,4-c]pyridin ylamino)methylpyridin-2(1H)-one 326b N N N 325a N N NH O a)3, xantphos Cs2CO3, dioxane N Br 100 °C, 2.5 h N Br 326a 326b N N N NH 199e AcO O N Pd(dppf)Cl2, N N K3PO4, NaOAc CH3CN, H2O, O N N2, 100 °C 326c A 100-mL round-bottomed flask equipped with a magnetic stirrer and a reflux 5 condenser was charged with 5-bromo-2,3-dimethyl-2H-pyrazolo[3,4-c]pyridine 326a from Example 325 (452 mg, 2.0 mmol), obromomethylpyridin-2(1H)-one 325a (400 mg, 2.0 mmol), cesium carbonate (1.3 g, 4.0 mmol), and 1,4-dioxane (10 mL). After bubbling nitrogen through the suspension for 5 s, BINAP (124 mg, 0.2 mmol) and tris(dibenzylideneacetone)dipalladium(0) (140 mg, 0.2 mmol) were added. The system was 10 subjected to three cycles of vacuum/nitrogen flush and heated at reflux for 2.5 h. It was then cooled to room temperature and filtered. The solid was washed with dichloromethane (3 X 10 mL). The combined filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with petroleum ether/ethyl acetate (2/1 to 100% ethyl acetate) to afford 326b (160 mg, 23%) as a yellow solid. MS-ESI: [M+H]+ 15 348.3 Example 326c {4-[5-({2,3-Dimethyl-2H-pyrazolo[3,4-c]pyridinyl}amino)- 1-methyloxo-1,6-dihydropyridinyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}methyl Acetate 326c A 50-mL round-bottomed flask equipped with a reflux condenser was charged with 20 326b (160 mg, 0.46 mmol), cetoxy)methyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (300 mg, 467 0.69 mmol), K3PO4 (195 mg, 0.92 mmol), sodium acetate (75 mg, 0.92 mmol), 1,1’- bis(diphenylphosphino) ferrocenedichloropalladium(II) (42 mg, 0.046 mmol), acetonitrile (10 mL), and water (6 drops). The system was subjected to three cycles of /nitrogen flush and heated at 100°C under N2 protection for 1.5 h. LCMS Analysis showed complete 5 conversion to the desired product. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue was diluted with dichloromethane (50 mL) and water (50 mL). The aqueous layer was separated and extracted with dichloromethane (3 × 20 mL). The combined extract was dried over , filtered, and concentrated under reduced pressure. The dark residue was purified by gel-silica column 10 chromatography eluting with 60:1 dichloromethane/methanol to afford 326c (130 mg, 45%) as a black solid. MS-ESI: [M+H]+ 621.3 Example 326 3-[4-[5-[(2,3-dimethylpyrazolo[3,4-c]pyridinyl)amino]methyl oxopyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 326 15 To a solution of 326c (130 mg, 0.21mmol) in THF/i-propanol /water(4/2/1 mL) was added lithium hydroxide (50 mg, 2.0 mmol) at room ature. After the reaction was stirred for 3h, LCMS indicated the on was complete. Then the mixture was poured into water (30 mL) and extracted with dichloromethane (3 X 30 mL). The combined organic layer was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced 20 pressure. The e solid was ed by reverse-phase prep-HPLC (A: 1‰NH4HCO3/water, B: acetonitrile) to afford 326 (60 mg, 50%) as a white solid. MS-ESI: [M+H]+ 579.3. 1H NMR (500 MHz, DMSO-d6) δ 8.86 (s, 1H), 8.48 (d, J = 5.0 Hz, 1H), 8.06 (d, J = 1.5Hz, 1H), 7.99 (s, 1H), 7.47 (d, J = 2.0 Hz, 1H), 7.41 (s, 1H), 7.38 (d, J = 5.0 Hz,1H), 6.57 (s, 1H), 5.13 (t, J = 5.0 Hz, 1H), 4.50-4.46 (m, 2H), 4.24-4.19 (m, 3H), 4.09 (s, 25 3H), 3.86-3.85 (m, 1H), 3.62 (s, 3H), 2.62-2.53 (m, overlap, 5H), 2.43 (s, 2H), 1.22 (s, 6H) Example 327a 5-(2-Methoxyethyl)nitro-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazine 327a O N N N N+O -O 468 To a solution of 2-nitro-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine (190 mg, 1.13 mmol) 209a in acetonitrile (10 mL) was added K2CO3 (311.9 mg, 2.26 mmol) and 1-bromo- 2-methoxyethane (188.3 mg, 1.36 mmol). The on mixture was heated at 80oC for 17 h under microwave irradiation. Analysis of the reaction mixture by LCMS showed complete 5 conversion to the d product. The e was cooled to room temperature and filtered.
The te was concentrated under reduced pressure to afford 327a as a white solid (230 mg, 90%), which was used in the next step without r purification. MS-ESI: [M+H]+ 227.0 Example 327b 5-(2-Methoxyethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin- 2-amine 327b 10 To a solution of 327a (286 mg, 1.26 mmol) in methanol (10 mL) was added Pd/C (28.6 mg). The system was evacuated and then refilled with H2. After stirring at room temperature for 2 h, the mixture was filtered. The filtrate was concentrated under reduced pressure to afford 327b as a yellow solid (240 mg, 97%), which was used in the next step 15 without further purification. MS-ESI: [M+H]+ 197.0 Example 327c bromomethylpyridin-2(1H)-one 327c 6-Methyl-pyridinol (10.9 g, 0.10 mol) was suspended in anhydrous dichloromethane (300 mL) and stirred at ambient temperature. Under cooling with an ice/water cooling bath, N-bromosuccinimide (NBS) (11.4 g, 0.20 mol) was added slowly 20 portion-wise over a time al of 5 minutes. The suspension was stirred at reflux for 2 hours. Thereafter, the suspension was ed. The filter cake was thoroughly washed with methanol and dried in vacuo to afford 327c as a white solid (22.7 g, 85%). MS-ESI: [M+H]+ 266.
Example 327d 3,5-Dibromo-1,6-dimethylpyridin-2(1H)-one 327d 25 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer was charged with DMF (50 mL), 327c (10.0 g, 37.5 mmol), CH3I (5.3 g, 37.5 mmol), and K2CO3 (7.8 g, 56.2 mmol). The mixture was stirred at room temperature for 5 h. Water (100 mL) was 469 added and the resulting white solid was collected to afford 327d (8.2 g, 78%) as a white solid.
MS-ESI: [M+H]+ 280.
Example 327e 5-Bromo(5-(2-methoxyethyl)-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)-1,6-dimethylpyridin-2(1H)-one 327e 5 A 100-mL round-bottomed flask equipped with a reflux condenser was charged with 327b (392 mg, 2.0 mmol), 327d (562 mg, 2.0 mmol), cesium carbonate (1.30 g, 4.0 mmol), and 1,4-dioxane (20 mL). After bubbling nitrogen through the suspension for 10 minutes, xantphos (115 mg, 0.20 mmol) and ibenzylideneacetone)dipalladium(0) (92 mg, 0.10 mmol) were added. The system was subjected to three cycles of vacuum/nitrogen flush and 10 heated at reflux for 5 h. It was then cooled to room temperature and filtered. The solid was washed with dichloromethane (2 X 15 mL). The combined filtrate was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (80:1 to 30:1) to afford 327e (490 mg, 62%) as a yellow solid.
MS-ESI: [M+H]+ 396.2 15 Example 327f (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}(5-{[5-(2-methoxyethyl)-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin yl]amino}-1,2-dimethyloxo-1,6-dihydropyridinyl)pyridinyl)methyl Acetate 327f A 25-mL single-neck bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 327e (158 mg, 0.40 mmol), {3-[(acetyloxy)methyl] 20 {4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridin yl}boronic acid 199e (159 mg, 0.40 mmol), K3PO4 (170 mg, 0.80 mmol), sodium acetate (66 mg, 0.80 mmol), Pd(dppf)Cl2 (15 mg, 0.020 mmol), and acetonitrile/water (7/0.5 mL). After three cycles of vacuum/N2 flush, the mixture was heated at 95oC for 1 h. LCMS analysis showed complete sion to the desired t. The on e was cooled to room 25 temperature, and diluted with dichloromethane (50 mL) and water (30 mL). The water layer was extracted with dichloromethane (2 × 30 mL). The combined organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. The dark residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (80:1 to 30:1) to afford 327f (120 mg, 45%) as a yellow solid. MS-ESI: [M+H]+ 668.8 30 Example 327 3-[3-(hydroxymethyl)[5-[[5-(2-methoxyethyl)-6,7-dihydro-4H- pyrazolo[1,5-a]pyrazinyl]amino]-1,2-dimethyloxopyridyl]pyridyl]-7,7-dimethyl- 1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 327 To a solution of 327f (120 mg, 0.18 mmol) in THF/ anol /water(6/4/3 mL) was added m hydroxide (22 mg, 0.90 mmol). The mixture was d at room temperature 470 for 1 h. The mixture was concentrated under reduced pressure and the residue was diluted with water (15mL). It was then extracted with ethyl acetate (3 x 20 mL). The combined organic layer was dried over sodium e and concentrated under pressure. The residue was purified by reverse-phase prep-HPLC to afford 327 as a white solid (55 mg, 49%). MS- 5 ESI: [M+H]+ 626.9. 1H NMR (500 MHz, CDCl3) δ 8.49 (d, J = 5.0 Hz, 1H), 7.49 (s, 1H), 7.32 (s, 1H), 7.15 (d, J = 4.5 Hz, 1H), 6.82 (s, 1H), 5.61 (bs, 1H), 4.53-4.45 (m, 3H), 4.26- 4.16 (m, 3H), 4.03-3.97 (m, 3H), 3.71-3.69 (m, 5H, overlap), 3.58 (t, J = 5.5 Hz, 2H), 3.39 (s, 3H), 2.98 (t, J = 5.0 Hz, 2H), 2.77 (t, J = 5.0 Hz, 2H), 2.60-2.57 (m, 2H), 2.53 (s, 2H), 2.17 (s, 3H), 1.29 (s, 6H). 10 Example 328a 2-Methoxyethyl)-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazinylamino)methyl(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin- 2(1H)-one 328a A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 15 reflux condenser was d with 5-bromo(5-(2-methoxyethyl)-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinylamino)methylpyridin-2(1H)-one 296f (330 mg, 0.86 mmol), Pin2B2 (329mg, 1.30 mmol), Pd2(dba)3 (40 mg, 0.043 mmol), X-phos (41 mg, 0.086 mmol), ium acetate (169 mg, 1.726 mmol), and dioxane (10 mL). After three cycles of vacuum/N2 flush, the mixture was heated at 70oC for 2 h. Analysis of the reaction mixture by 20 LCMS showed complete conversion to the desired product. It was cooled to room temperature and filtered. The te was trated under reduced pressure. The residue was washed with petroleum ether to afford 328a as a dark oil (240 mg, 80%), which was used in the next step without further purification. MS-ESI: [M+H]+ 348.3 Example 328b 3-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 25 2(6),7-dienyl}(5-{[5-(2-methoxyethyl)-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin yl]amino}methyloxo-1,6-dihydropyridinyl)pyridinecarbaldehyde 328b 471 A sealed tube ed with a magnetic stirrer was charged with 3-bromo{4,4- dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyri-dine carbaldehyde 107f (100 mg, 0.26 mmol), 328a (110 mg, 0.26 mmol), Pd(dppf)Cl2 (10 mg, 0.026mmol), sodium acetate (50 mg, 0.50 mmol), K3PO4 (100 mg, 0.50 mmol), and 5 acetonitrile / water (5 mL/1mL). After three cycles of /nitrogen flush, the mixture was heated at 100 oC for 2 h. It was then filtered and the filtrate was evaporated under d pressure. The residue was ed by -gel column chromatography eluting with 10:1 dichloromethane/methanol to afford 328b (50 mg, 32%) as a brown solid. MS-ESI: [M+H]+ 611.3. 10 Example 328 3-[4-(hydroxymethyl)[5-[[5-(2-methoxyethyl)-6,7-dihydro-4H- pyrazolo[1,5-a]pyrazinyl]amino]methyloxopyridyl]pyridyl]-7,7-dimethyl- 1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 328 A mixture of 328b (50 mg, 0.08 mmol) and NaBH4 (8.0 mg, 0.20 mmol) in methanol (4 mL) was stirred at 25 oC for 0.5 h. The reaction mixture was quenched with water (10 mL) 15 and evaporated under reduced pressure. The residue was added extracted with dichloromethane (2 X 10 mL). The combined extract was concentrated under reduced pressure and the e was purified by reverse-phase prep-HPLC to afford 328 (13 mg, 25%) as a pale yellow solid. MS-ESI: [M+H]+ 613.3. 1H NMR (500 MHz, CDCl3) δ 8.62 (s, 1H), 8.48 (s, 1H), 7.92 (d, J = 2.0 Hz, 1H), 7.38 (s, 1H), 7.31 (d, J = 2.0 Hz, 1H), 6.82 (s, 1H), 20 5.67 (s, 1H), 4.64-4.62 (m, 1H), 4.57-4.55 (m, 1H), 4.38-4.34 (m, 1H), 4.22-4.17 (m, 3H), 4.05-4.02 (m, 2H), 3.99-3.96 (m, 1H), 3.71-3.70 (m, 2H), 3.69 (s, 3H), 3.57 (t, J = 5.0 Hz, 2H), 3.37 (s, 3H), 2.99 (t, J = 5.0 Hz, 2H), 2.77 (t, J = 5.0 Hz, 2H), 2.56 (s, 2H), 2.51 (s, 2H), 1.27 (s, 6H).
Example 329a 2-Chloro{4,4-dimethyloxo-1,10- 25 diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinecarbaldehyde 329a 472 A 100-mL round-bottomed flask equipped with a reflux condenser was charged with 4,4-dimethyl-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienone 107e (612 mg, 3.0 mmol), 4-bromochloronicotinaldehyde (2.0 g, 9.0 mmol), Pd2(dba)3 (275 mg, 0.30 mmol), XantPhos (347 mg, 0.60 mmol), cesium carbonate (1.95 g, 6.0 mmol), and 1,4-dioxane (30 5 mL). After three cycles of vacuum/nitrogen flush, the mixture was heated at 97oC overnight.
It was then cooled to room temperature and filtered. The te was concentrated under reduced re and the resulting residue was purified by -gel column chromatography g with 1:2 ethyl acetate/petroleum ether to afford 329a as a yellow solid (660 mg, 65%).
MS-ESI: [M+H]+ 344.1. 10 Example 329b 4-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}(5-{[5-(2-methoxyethyl)-4H,5H,6H,7H-pyrazolo[1,5-a]pyrazin yl]amino}methyloxo-1,6-dihydropyridinyl)pyridinecarbaldehyde 329b A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux ser was charged with 329a (100 mg, 0.30 mmol), 3-(5-(2-methoxyethyl)- 15 4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinylamino)methyl(4,4,5,5-tetramethyl-1,3,2- orolanyl)pyridin-2(1H)-one 328a (257 mg, 0.60 mmol), Pd(dppf)Cl2 (25 mg, 0.030 mmol), K3PO4 (127 mg, 0.60 mmol), sodium acetate (49 mg, 0.60 mmol), water (0.50 mL), and THF (10 mL). After three cycles of vacuum/nitrogen flush, the mixture was heated at reflux for 1 h. It was then cooled to room temperature and filtered. The filtrate was 20 concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 329b as a brown solid (60 mg, 34%). MS-ESI: [M+H]+ 611.3.
Example 329 3-[3-(hydroxymethyl)[5-[[5-(2-methoxyethyl)-6,7-dihydro-4H- pyrazolo[1,5-a]pyrazinyl]amino]methyloxopyridyl]pyridyl]-7,7-dimethyl- 25 1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 329 A mixture of 329b (50 mg, 0.080 mmol) and NaBH4 (9.1 mg, 0.24 mmol) in methanol (5 mL) was stirred at room temperature for 10 min. The mixture was quenched with water (10 mL) and evaporated under d pressure. The residue was extracted with dichloromethane (3 X 10 m). The combined extract was concentrated under reduced pressure 30 and the residue was purified by e-phase PLC to afford 329 (15 mg, 30%) as a yellow solid. MS-ESI: [M+H]+ 613.3. 1H NMR (500 MHz, CDCl3) δ 8.66 (d, J = 4.5 Hz, 1H), 8.13 (s, 1H), 7.63 (s, 1H), 7.35 (s, 1H), 7.12 (d, J = 5.0 Hz, 1H), 6.83 (s, 1H), 5.71 (s, 1H), 4.67-4.63 (m, 1H), 4.49-4.43 (m, 1H), 4.24-4.23 (m, 2H), 4.19-4.17 (m, 1H), 4.06-4.04 (m, 473 2H), 4.01-3.97 (m, 1H), 3.74-3.71 (m, 2H), 3.70 (s, 3H), 3.59-3.55 (m, 2H), 3.38 (s, 3H), 3.00 (t, J = 5.0 Hz, 2H), 2.77 (t, J = 5.0 Hz, 2H), 2.56 (s, 2H), 2.51 (s, 2H), 1.27 (s, 6H).
Example 330a 10-[4-Chloro(hydroxymethyl)pyridinyl]-4,4-dimethyl thiaazatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienone 330a 5 A mixture of 4-chloro{4,4-dimethyloxothiaazatricyclo[6.4.0.02,6]dodeca- 1(8),2(6)-dienyl}pyridinecarbaldehyde 109a (1.2 g, 3.3 mmol), NaBH4 (228 mg, 6.0 mmol), and methanol (10 mL) was d at 0oC for 0.5 h. Then the reaction mixture was quenched with water (10 mL) and concentrated under reduced pressure. The residue was 10 ted with dichloromethane (2 X 15 mL). The combined dichloromethane extract was concentrated under reduced pressure to afford 330a as a pale yellow solid (1.0 g, 84%). MSESI : [M+H]+ 362.9 Example 330b (4-Chloro{4,4-dimethyloxothia azatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienyl}pyridinyl)methyl Acetate 330b 15 A mixture of 330a (1.0 g, 2.76 mmol), triethylamine (610 mg, 6.0 mmol), and acetic anhydride (5 mL) was stirred at 25oC for 2 h. Then the reaction mixture was quenched with water (10 mL) and the pH was adjusted to around 8 with NaHCO3 (aq.). The mixture was extracted with dichloromethane (2 X 15 mL). The combined romethane extract was concentrated under d pressure and the residue was purified by -gel column 20 chromatography eluting with 1:1 ethyl acetate /petroleum ether to afford 330b as a pale yellow solid (1.0 g, 90%). MS-ESI: [M+H]+ 405.2 474 Example 330c (2-{4,4-Dimethyloxothia azatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienyl}(4,4,5,5-tetramethyl-1,3,2- dioxaborolanyl)pyridinyl)methyl Acetate 330c A 50-mL bottomed flask equipped with a magnetic stirrer and a reflux 5 condenser was charged with 330b (1.0 g, 2.47 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'- bi(1,3,2-dioxaborolane) (1.87 g, 7.40 mmol), Pd(dppf)Cl2 (100 mg, 0.13 mmol), X-phos (125 mg, 0. 25 mmol), potassium acetate (500 mg, 5.0 mmol) and 1,4-dioxane (10 mL). After three cycles of vacuum/nitrogen flush, the mixture was heated at 65 oC for 4 h. It was then ed and the filtrate was evaporated under reduced re to afford 330c (1.0 g, 98%) as 10 a brown oil without further purification. MS-ESI: [M+H]+ 415.2.
Example 330d (3-Nitro-1H-pyrazolyl)methanol 330d A mixture of 3-nitro-1H-pyrazolecarboxylic acid (4.71 g, 30 mmol), F (75 mL, 1 mol/L, 75 mmol) was stirred at 60oC for 2 h. The mixture was cooled to room temperature and 4M HCl (19 mL, 75 mmol) was added. It was d at 70oC for 2 h. After 15 cooling to room temperature, the mixture was concentrated under reduced pressure. The e was partitioned between ethyl acetate and brine (100:100 mL). The aqueous phase was extract with ethyl acetate (4 X 50 mL). The combined organic layer was dried on Na2SO4 and evaporated under reduced pressure. The residue was purified by silica-gel column tography eluting with petroleum ether/ethyl acetate (5:1 to 1:1) to afford 330d (3.5 g, 20 79%) as a white solid. MS-ESI: [M+H]+ 144.2 Example 330e 1-(5-(Hydroxymethyl)nitro-1H-pyrazolyl) propanol 330e A sealed tube was charged with 330d (2.145 g, 15 mmol), Cs2CO3 (978 mg, 3.0 mmol), and 2,2-dimethyloxirane (15 mL). The mixture was stirred at 70oC for 3 h. After 25 cooling to room temperature, the mixture was concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with petroleum ethyl acetate (5:1 to 1:1) to afford 330e (1.2 g, 38%) as a white solid. MS-ESI: [M+H]+ 216.2 Example 330f methylnitro-6,7-dihydro-4H-pyrazolo[5,1- c][1,4]oxazine 330f 30 To a solution of 330e (1.1 g, 5.1 mmol) in DMF (10 mL), was added NaH (60 percent sion in mineral oil, 246 mg, 6.14 mmol) at 0 oC. The resulting suspension was stirred for 30 min, followed by the addition of p-toluenesulfonyl chloride (1169 mg, 6.14 mmol).
The mixture was stirred at 60oC overnight. After cooling to room temperature, saturated ammonium chloride solution was added and the mixture was extracted with dichloromethane. 475 The combined organic layer was washed with brine, dried over Na2SO4, and evaporated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with eum ether/ethyl acetate gradient (9:1 to 2:1) to afford 330f (228 mg, 22%).
MS-ESI: [M+H]+ 198.3 5 Example 330g 6,6-Dimethyl-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin amine 330g A 50-mL single-neck round-bottomed flask was purged with nitrogen and charged with 330f (0.21 g, 1.25 mmol), 10% palladium on carbon (50% wet, 125 mg), and methanol (10 mL). The mixture was evacuated, charged with en gas, and stirred at room 10 temperature for 2 h. The hydrogen was then evacuated and nitrogen was charged into the flask. The catalyst was removed by filtration through a pad of ® and the filtrate was concentrated under reduced pressure to afford 330g (167 mg, 93%). MS-ESI: [M+H]+ 168.1 Example 330h 6-Chloro(6,6-dimethyl-6,7-dihydro-4H-pyrazolo[5,1- c][1,4]oxazin ylamino)methylpyridazin-3(2H)-one 330h Br O O O N Cl N N N N NH N NH2 Pd2(dba)3, xantphos O Cs2CO3, e 100 °C N 330g Cl N 330h O N N NH OAc O 330c S N N N PdCl2(dppf), O N AcONa, CH3CN, 95 °C, 3h 15 330i A 50-mL single-neck bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 330g (250 mg, 1.5 mmol), 4-bromochloro pyridazin-3(2H)-one (669 mg, 3.0 mmol), Pd2(dba)3 (137 mg, 0.15 mmol), Xantphos (173 mg, 0.30 mmol), Cs2CO3 (978 mg, 3.0 mmol), and 1,4-dioxane (20 mL). After three 20 cycles of vacuum/argon flush, the mixture was heated at 100ºC for 3 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and 476 washed with ethyl acetate to afford 330h as a yellow solid (209 mg, 45%). MS-ESI: [M+H]+ 310.1.
Example 330i {4-[5-({6,6-Dimethyl-4H,6H,7H-pyrazolo[3,2-c][1,4]oxazin yl}amino)methyloxo-1,6-dihydropyridazinyl]{4,4-dimethyloxothia 5 azatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienyl}pyridinyl}methyl Acetate 330i A 100-mL single-neck bottomed flask equipped with a ic stirrer and reflux condenser was charged with 330h (133 mg, 0.43 mmol), 330c (178 mg, 0.43 mmol), sodium acetate (71 mg, 0.86 mmol), K3PO4 (182 mg, 0.86 mmol), Pd(dppf)Cl2 (35 mg, 0.043 mmol), acetonitrile (15 mL), and water (0.5 mL). After bubbling nitrogen through the 10 resulting mixture for 20 minutes, the reaction mixture was heated at 95 oC for 3 h. After this time the reaction was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue was purified with silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 330i as a yellow solid (69 mg, 25%).
MS-ESI: [M+H]+ 644.3. 15 Example 330 3-[4-[5-[(6,6-dimethyl-4,7-dihydropyrazolo[5,1-c][1,4]oxazin yl)amino]methyloxo-pyridazinyl](hydroxymethyl)pyridyl]-7,7-dimethyl- 1,2,6,8-tetrahydrocyclopenta[3,4]thieno[1,3-c]pyridinone 330 A mixture of 330i (69 mg, 0.11 mmol) and lithium hydroxide (10 mg, 0.42 mmol) in i-propanol/THF (1:1, 3.5 mL) and water (1 mL) was stirred at room temperature for 1 h. The 20 mixture was then trated under reduced pressure and the residue was d with water (10 mL). It was extracted with ethyl e (2 X 10 mL). The combined ethyl acetate extract was concentrated under reduced pressure and the residue was ed with reverse-phase PLC to afford 330 (30 mg, 47%). MS-ESI: [M+H]+ 602.5. 1H NMR (500 MHz, CDCl3) δ 8.56 (d, J = 5.0 Hz, 1H), 8.00 (s, 1H), 7.90 (s, 1H), 7.43 (d, J = 5.0 Hz, 1H), 5.94 (s, 25 1H), 4.82 (s, 2H), 4.60-4.58 (m, 2H), 4.38-4.36 (m, 2H), 3.89 (s, 3H), 3.89-3.87 (m, 3H), 3.02-2.93 (m, 2H), 2.79-2.75 (m, 2H), 2.59-2.54 (m, 2H), 1.37 (s, 6H), 1.28 (s, 6H).
Example 331a 5-Bromomethyl(3-methylisothiazolylamino)pyrazin- 2(1H)-one 331a 477 A sealed tube equipped with a magnetic stirrer was charged with ylisothiazol- 5-amine (170 mg, 1.5 mmol), 3,5-dibromomethylpyrazin-2(1H)-one (400 mg, 1.5 mmol), Pd(OAc)2 (84 mg, 0.375 mmol), BINAP (116 mg, 0.188 mmol), K2CO3 (450 mg, 4.5 mmol), 5 and 1,4-dioxane (4 mL). After three cycles of /nitrogen flush, the mixture was heated at 120oC in a sealed tube for 18 h. It was then cooled to room temperature and filtered. The filtrate was evaporated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (100:1 to 25:1) to afford 331a (220 mg, 50%) as a yellow solid. MS-ESI: [M+H]+ 301.0. 10 Example 331b (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}{4-methyl[(3-methyl-1,2-thiazolyl)amino]oxo-4,5- dihydropyrazinyl}pyridinyl)methyl Acetate 331b A 50-mL bottomed flask ed with a reflux condenser was charged with 331a (150 mg, 0.50 mmol), {3-[(acetoxy)methyl]{4,4-dimethyloxo-1,10- 15 diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (400 mg, 1.0 mmol), Pd(dppf)Cl2 (25 mg, 0.025 mmol), K3PO4 (220 mg, 1.0 mmol), sodium acetate trihydrate (136 mg, 1.0 mmol), acetonitrile (10 mL), and water (0.5 mL). The system was evacuated and refilled with N2. The reaction e was heated at 100 ºC for 1 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure 20 and the resulting residue was purified by silica-gel column chromatography g with dichloromethane/methanol (100/1 to 25/1) to afford 331b (200 mg, 70%) as a yellow solid.
MS-ESI: [M+H]+ 574.2.
Example 331 3-[3-(hydroxymethyl)[4-methyl[(3-methylisothiazol yl)amino]oxo-pyrazinyl]pyridyl]-7,7-dimethyl-1,2,6,8- 25 ydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 331 A mixture of 331b (120 mg, 0.21 mmol) and lithium hydroxide monohydrate (88 mg, 2.1 mmol) in THF/i-propanol (4:2, 6 mL) and water (2 mL) was stirred at 30 oC for 1 h. The mixture was evaporated under reduced pressure and diluted with water (10 mL). It was then 478 extracted with ethyl e (2 x 15 mL). The combined ethyl acetate extract was concentrated under reduced pressure and the residue was purified by reverse-phase prep- HPLC to afford 331 (50 mg, 45 %) as a white solid. MS-ESI: [M+H]+ 532.2. 1H NMR (500 MHz, CHCL3) δ 9.13 (s, 1H), 8.61 (d, J = 5 Hz, 1H), 8.37 (s, 1H), 7.99 (d, J = 5.0 Hz, 1H), 5 6.87 (s, 1H), 6.74 (s, 1H), 5.32-5.39 (m, 1H), .75 (m, 1H), 4.58-4.56 (m, 1H), 4.32- 4.37 (m, 1H), 4.21-4.18 (m, 2H), 3.96-3.94 (m, 1H), 3.72 (s, 3H), 2.61-2.58 (m, 2H), 2.54 (s, 2H), 2.48 (s, 3H), 1.30 (s, 6H).
Example 332a 5-Bromo(5-ethyl-1,3,4-thiadiazolylamino) methylpyridin-2(1H)-one 332a 10 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 5-ethyl-1,3,4-thiadiazolamine (500 mg, 3.88 mmol), 3,5-dibromomethylpyridin-2(1H)-one (1.55 g, 5.81 mmol), Pd2(dba)3 (357 mg, 0.39 mmol), XantPhos (451 mg, 0.78 mmol), Cs2CO3 (2.5 g, 7.67 mmol), and 1,4-dioxane (40 mL). 15 The system was ted to three cycles of vacuum/nitrogen flush and heated at 100oC for 2 h. The mixture was cooled to 90oC and filtered. The te was cooled in an ice-water bath and then filtered again to afford 332a (574 mg, 47%) as a white solid. MS-ESI: [M+H]+ 315.1 Example 332b (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 20 2(6),7-dienyl}{5-[(5-ethyl-1,3,4-thiadiazolyl)amino]methyloxo-1,6- dihydropyridinyl}pyridinyl)methyl e 332b A 100-mL round-bottomed flask equipped with a reflux condenser was charged with 332a (200 mg, 0.63 mmol), {3-[(acetoxy)methyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (754 mg, 25 1.89 mmol), PdCl2(dppf) (51 mg, 0.063 mmol), K3PO4 (267 mg, 1.26 mmol), CH3COONa (103 mg, 1.26 mmol), itrile (15 mL), and water (0.5 mL). After bubbling nitrogen through the resulting mixture for 20 minutes, it was heated at 100oC under a nitrogen 479 atmosphere for 2 h. The resulting mixture was cooled to room temperature and filtered. The filtrate was evaporated under reduced pressure and the residue was purified by silica-gel column chromatography g with 50:1 dichloromethane/methanol to afford 332b as a brown solid (178 mg, 48%). MS-ESI: [M+H]+ 588.2 5 Example 332 3-[4-[5-[(5-ethyl-1,3,4-thiadiazolyl)amino]methyloxo pyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 332 A mixture of 332b (158 mg, 0.27 mmol) and lithium hydroxide (19 mg, 0.81 mmol) in i-propanol/THF/water (9 mL /6 mL /6 mL) was d at room temperature for 0.5 h. The 10 mixture was evaporated under reduced pressure and the residue was extracted with dichloromethane (3 X 20 mL). The combined dichloromethane t was concentrated under reduced pressure and the residue was purified by reverse-phase prep-HPLC to afford 332 as a white solid (80 mg, 54%). MS-ESI: [M+H]+ 546.2. 1H NMR (500 MHz, DMSO-d6) δ 10.20 (s, 1H), 8.60 (d, J = 2.0 Hz, 1H), 8.49 (d, J = 5.0 Hz, 1H), 7.63 (d, J = 2.5 Hz, 1H), 15 7.33 (d, J = 5.0 Hz, 1H), 6.57 (s, 1H), 4.92 (t, J = 4.5 Hz, 1H), .39 (m, 2H), 4.25-4.19 (m, 3H), 3.87-3.85 (m, 1H), 3.61 (s, 3H), 2.92-2.88 (m, 2H), 2.58-2.53 (m, 2H), 2.43 (s, 2H), 1.27-1.22 (m, overlap, 9H). e 333a (2-{4,4-Dimethyloxothia azatricyclo[6.4.0.02,6]dodeca-1(8),2(6)-dienyl}{1-methyl[(1-methyl-1H-1,2,3- 20 triazolyl)amino]oxo-1,6-dihydropyridinyl}pyridinyl)methyl Acetate 333a A 50-mL round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with (2-{4,4-dimethyloxothiaazatricyclo[6.4.0.02,6]dodeca- 1(8),2(6)-dienyl}(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridinyl)methyl 25 acetate 330c (180 mg, 0.37 mmol), 5-bromomethyl(1-methyl-1H-1,2,3-triazol o)pyridin-2(1H)-one 292c (125 mg, 0.43 mmol), Pd(dppf)Cl2 (20 mg, 0.025 mmol), potassium acetate (80 mg, 0.80 mmol), K3PO4 (165 mg, 0.80 mmol), and acetonitrile/water (10 mL /1 mL). After three cycles of /nitrogen flush, the mixture was heated at 100oC 480 for 1 h. It was then filtered and the filtrate was evaporated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 20:1 romethane/methanol to afford 333a (150 mg, 71%) as a brown solid. MS-ESI: [M+H]+ 574.1 5 Example 333 3-[3-(hydroxymethyl)[1-methyl[(1-methyltriazolyl)amino] oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]thieno[1,3- dinone 333 A mixture of 333a (150 mg, 0.26 mmol) and lithium hydroxide hydrate (84 mg, 2.0 mmol) in THF (5 mL), i-propanol (5 mL) and water (1.5 mL) was d at 40oC for 0.5 h. 10 The mixture was evaporated under reduced pressure and diluted with water (10 mL). It was then extracted with dichloromethane (2 X 10 mL). The combined organic layer was concentrated under d pressure and the e was purified by reverse-phase prep- HPLC to afford 333 (52 mg, 38%) as a pale yellow solid. MS-ESI: [M+H]+ 532.3. 1H NMR (500 MHz, DMSO-d6) δ 8.48-8.47 (m, 1H), 8.28 (s, 1H), 7.78-7.77 (m, 2H), 7.42 (d, J = 2.5 15 Hz, 1H), 7.34-7.33 (m, 1H), 4.97-4.95 (t, J = 5.0 Hz, 1H), 4.43-4.41 (m, 2H), 4.17-4.16 (m, 1H), 3.99 (s, 3H), 3.94-3.92 (m, 1H), 3.59 (s, 3H), 3.04-3.02 (m, 1H), 2.90-2.89 (m, 1H), 2.77-2.75 (m, 2H), 2.56-2.54 (m, 2H), 1.23 (s, 3H), 1.22 (s, 3H).
Example 334 3-[4-[5-[(5-cyclopropyl-1,3,4-thiadiazolyl)amino]methyloxo pyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8- 20 tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 334 Following the procedures of Example 273, and tuting 5-cyclopropyl-1,3,4- thiadiazolamine for 2-amino pyridine gave 334 (8.7 mg, 22% yield). 1H NMR (400 MHz, DMSO-d6) δ 8.48 (d, J = 5.1 Hz, 1H), 8.20 (s, 1H), 8.10 (d, J = 2.4 Hz, 1H), 7.53 (d, J = 2.4 Hz, 1H), 7.44 (d, J = 5.1 Hz, 1H), 6.53 (s, 1H), 4.95 (t, J = 5.2 Hz, 1H), 4.55 – 4.49 (m, 1H), 25 4.27 – 4.23 (m, 3H), 3.78 (s, 2H), 3.51 (s, 3H), 3.48 (dt, J = 12.3, 5.2 Hz, 3H), 3.24 (s, 2H), 2.50 (d, J = 7.2 Hz, 2H), 2.33 (s, 2H), 1.85 – 1.82 (m, 3H), 1.23 (s, 6H). ES-MS m/z 531.3 [M+1].
Example 335a 5-Bromo(6,6-dimethyl-6,7-dihydro-4H-pyrazolo[5,1- c][1,4]oxazinylamino)methylpyridin-2(1H)-one 335a 481 O O N N N NH N NH O O O N N B Br Pin2B2, Pd(dppf)Cl2 O KOAc, dioxane 335a 90 oC, 4 h 335b O N N NH O O 282i N S N N PdCl2(dppf), O N K3PO4,NaOAc MeCN, 95 °C, 1 h, H2O 335c A 100-mL bottomed flask equipped with a reflux condenser was d with 1,4-dioxane (10 mL), 6,6-dimethyl-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazinamine 330g (167 mg, 1.0 mmol), 3,5-dibromomethylpyridin-2(1H)-one (320 mg, 1.2 mmol), 5 Pd2(dba)3 (91 mg, 0.10 mmol), XantPhos (116 mg, 0.20 mmol), and cesium carbonate (652 mg, 2.0 mmol). After three cycles of vacuum/argon flush, the mixture was heated at 100 oC for 3 h. It was then filtered and the te was evaporated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 100:1 dichloromethane/methanol to afford 335a (210 mg, 60%) as a yellow solid. MS-ESI: [M+H]+ 10 352.9 e 335b 3-(6,6-Dimethyl-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin ylamino)methyl(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 335b To a mixture of 335a (160 mg, 0.45 mmol) and 4,4,4',4',5,5,5',5'-octamethyl-2,2'- bi(1,3,2-dioxaborolane) (572 g, 2.25 mmol) in dioxane (20 mL) was added dppf) (36.8 15 mg, 0.045 mmol) and potassium acetate (88.2 mg, 0.90 mmol ). After three cycles of vacuum/nitrogen flush, the mixture was stirred at 90℃ for 4 h under nitrogen atmosphere. It was then filtered and the filtrate was evaporated under reduced pressure to afford 335b, which was used in the next step without further purification. MS-ESI: [M+H]+401.3. 482 Example 335c 4-[5-({6,6-Dimethyl-4H,6H,7H-pyrazolo[3,2-c][1,4]oxazin yl}amino)methyloxo-1,6-dihydropyridinyl]{4,4-dimethyloxothia-10,11- diazatricyclo[6.4.0.02,6]dodeca-1(8),2(6),11-trienyl}pyridinecarbaldehyde 335c A 50-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 5 reflux condenser was charged with 335b (240 mg, 0.60 mmol), 4-chloro{4,4-dimethyl thia-10,11-diazatricyclo[6.4.0.02,6]dodeca-1(8),2(6),11-trienyl}pyridine carbaldehyde 282i (107.7 mg, 0.30 mmol), Pd(dppf)Cl2 (24.5 mg, 0.030 mmol), K3PO4 (127.2 mg, 0.60 mmol), sodium acetate (49.2 mg, 0.60 mmol), water (0.5 mL), and itrile (10 mL). After three cycles of vacuum/nitrogen flush, the mixture was heated at 10 95oC for 1 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced re and the resulting residue was purified by silica-gel column chromatography eluting with 30:1 dichloromethane/methanol to afford 335c as a brown solid (60 mg, 22%, two steps). MS-ESI: [M+H]+ 598.2.
Example 335 3-[4-[5-[(6,6-dimethyl-4,7-dihydropyrazolo[5,1-c][1,4]oxazin 15 yl)amino]methyloxopyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl-6,8- dihydrocyclopenta[3,4]thieno[1,3-d]pyridazinone 335 A mixture of 335c (50 mg, 0.080 mmol) and NaBH4 (9.1 mg, 0.24 mmol) in methanol (5 mL) was stirred at room temperature for 10 min. The mixture was quenched with water (10 mL) and evaporated under reduced pressure. The residue was ted with 20 dichloromethane (3 X 10 mL). The combined extract was concentrated under d pressure and the residue was purified by reverse-phase PLC to afford 335 (15 mg, 30%) as a yellow solid. MS-ESI: [M+H]+ 600.2. 1H NMR (500 MHz, DMSO-d6) δ 8.56 (d, J = 5.0 Hz, 1H), 8.46 (s, 1H), 8.31 (s, 1H), 8.07 (d, J = 2.0 Hz, 1H), 7.52 (d, J = 5.0 Hz, 1H), 7.40 (d, J = 2.5 Hz, 1H), 5.94 (s, 1H), 4.85-4.83 (m, 1H), 4.72 (s, 2H), 4.38-4.37 (m, 2H), 25 3.79-3.78 (m, 2H), 3.3 (s, 3H), 2.92-2.91 (m, 2H), 2.81 (s, 2H), 1.28 (s, 6H), 1.25 (s, 6H).
Example 336a 5-(Methoxymethyl)methylnitro-1H-pyrazole 336a A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 5-(bromomethyl)methylnitro-1H-pyrazole (8.8 g, 40 mmol), sodium ide (4.3 g, 80 mmol), and methanol (50 mL). The reaction mixture 30 was heated at reflux for 2 h. After this time the reaction was cooled to room temperature and concentrated under reduced pressure. The residue was partitioned between ethyl acetate (60 mL) and water (60 mL). The s layer was ted and extracted with ethyl acetate (2 X 50 mL). The combined organic layer was washed with brine (50 mL) and dried over 483 sodium sulfate. The drying agent was removed by tion and the filtrate was concentrated under d re to afford 336a as a yellow oil (6.1 g, 90%). MS-ESI: [M+H]+ 172.
Example 336b 5-(Methoxymethyl)methyl-1H-pyrazolamine 336b A 250-mL single-neck round-bottomed flask equipped with a magnetic stirrer was 5 charged with 336a (4.0 g, 23 mmol), Pd/C (1.0 g), and ethanol (100 mL). The mixture was hydrogenated at room temperature for 15 h. It was then filtered and the filtrate was trated under reduced pressure to afford 336b as a yellow oil (3.3 g, 99%), which was used in the next step without further purification. MS-ESI: [M+H]+ 142.
Example 336c 5-Bromo(5-(methoxymethyl)methyl-1H-pyrazol 10 ylamino)methylpyridin-2(1H)-one 336c O N HN N O N Br Following the procedure in Example 335a, and starting with 335b (1.7 g, 12 mmol) and 3,5-dibromomethylpyridin-2(1H)-one (3.2 g, 12 mmol) ed 336c as a yellow solid (2.8 g, 71%). MS-ESI: [M+H]+ 327. 1H NMR (500 MHz, CDCl3) δ 7.86 (d, J = 2.5 Hz, 15 1H), 7.38 (s, 1H), 6.88 (d, J = 2.5 Hz, 1H), 5.86 (s, 1H), 4.41 (s, 2H), 3.82 (s, 3H), 3.58 (s, 3H), 3.36 (s, 3H).
Example 336d 3-(5-(Methoxymethyl)methyl-1H-pyrazolylamino) methyl(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)pyridin-2(1H)-one 336d A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a 20 reflux condenser was charged with 336c (600 mg, 1.83 mmol), ,4',5,5,5',5'- octamethyl- 2,2'-bi(1,3,2-dioxaborolane) (1.39 g, 5.49 mmol), Pd2(dba)3 (183 mg, 0.20 mmol), X-phos (190 mg, 0.40 mmol), porassium e (392 mg, 4.0 mmol), and 1,4-dioxane (30 mL). After three cycles of vacuum/nitrogen flush, the mixture was heated at 85ºC for 3 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure 25 to afford crude 336d as a black oil (400 mg, 75%), which was used in the next step without purification. MS-ESI: [M+H]+ 293.1 Example 336e 4-(5-(5-(Methoxymethyl)methyl-1H-pyrazolylamino) methyloxo-1,6-dihydropyridinyl)(1-oxo-3,4,6,7,8,9-hexahydropyrido[3,4- b]indolizin-2(1H)-yl)nicotinaldehyde 336e 484 A 50-mL round-bottomed flask equipped with a reflux condenser was charged with 336d (368 mg, 0.98 mmol), 4-chloro(1-oxo-3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizin- 2(1H)-yl)nicotinaldehyde 139a (270 mg, 0.82 mmol), dppf) (60 mg, 0.082 mmol), 5 K3PO4 (348 mg, 1.64 mmol)), sodium acetate (135 mg, 1.65 mmol), acetonitrile (15 mL), and water (0.5 mL). The system was subjected to three cycles of /nitrogen flush and heated at 95oC for 3 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the residue was ed by silica-gel column tography eluting with 20:1 ethyl acetate/methanol to afford 336e (100 mg, 22%). MS- 10 ESI: [M+H]+ 542.2 Example 336 2-[3-(hydroxymethyl)[5-[[5-(methoxymethyl)methyl-pyrazol yl]amino]methyloxopyridyl]pyridyl]-3,4,6,7,8,9-hexahydropyrido[3,4- b]indolizinone 336 To a solution of 336e (100 mg, 0.18 mmol ) in methanol (10 mL) was added NaBH4 15 (41 mg, 1.08 mmol). The mixture was stirred at room temperature for 1 h and LCMS showed the starting material had disappeared. The reaction was quenched with 1.0 M HCl solution (10 mL) and evaporated under reduced pressure until most of methanol was distilled. The residue was extracted with dichloromethane (3 X 15 mL). The combined organic layer was dried with Na2SO4 and evaporated under reduced pressure. The residue was purified by 20 reverse-phase prep-HPLC to afford 336 as a white solid (41 mg, 41%). MS-ESI: [M+H]+ 544.2. 1H NMR (500 MHz, CDCl3) δ 8.49 (d, J = 5.0 Hz, 1H), 7.96 (d, J = 2.5 Hz, 1H), 7.73 (d, J = 2.5 Hz, 1H), 7.41 (s, 1H), 7.33 (d, J = 5.0 Hz, 1H), 6.33 (s, 1H), 5.95 (s, 1H), 4.99- 4.96 (m, 1H), 4.67-4.64 (m, 1H), 4.42-4.41 (m, 3H), .26 (m, 1H), 3.98-3.91 (m, 1H), 3.88-3.82 (m, 2H), 3.79 (s, 3H), 3.71 (s, 3H), 3.37 (s, 3H), 3.06-2.91 (m, 2H), 2.87-2.79 (m, 25 2H), 2.08-2.01 (m, 2H), 1.91-1.86 (m, 2H).
Example 337a 1,2-Dimethylnitro-1H-imidazole 337a 485 To a e of 2-methylnitro-1H-imidazole (10.0 g, 78.7 mmol) and K2CO3 (21.7 g, 160 mmol) in DMF (80 mL) was added CH3I (13.4 g, 94 mmol) dropwise while stirring at room temperature. The mixture was stirred for 2 h. Water (200 mL) was then added to the 5 mixture. The resulting suspension was filtered, washed with water, and dried in vacuo to afford 337a as a white solid (5.0 g, 45%). MS-ESI: [M+H]+ 142.1.
Example 337b tert-Butyl 1,2-Dimethyl-1H-imidazolylcarbamate 337b A 100-mL -neck round-bottomed flask was purged with nitrogen and d with 337a (2.0 g, 14.1 mmol), 10% palladium on carbon (50% wet, 400 mg), (Boc)2O (9.22 g, 10 43.3 mmol), triethylamine (2.85 g, 28.2 mmol), and ethanol (20 mL). The e was evacuated, charged with hydrogen gas, and stirred at room temperature for 5 h. The hydrogen was then evacuated and nitrogen was charged into the flask. The catalyst was removed by filtration through a pad of CELITE® and the filtrate was concentrated under reduced pressure.
The residue was purified by silica-gel column chromatography eluting with 40:1 15 dichloromethane/methanol to afford 337b (1.2 g, 40%) as a brown solid. MS-ESI: [M+H]+ 212.1 Example 337c 1,2-Dimethyl-1H-imidazolamine hloride 337c To a solution of 337b (1.2 g, 5.68 mmol) in dichloromethane (5.0 mL) was added 3M HCl in dioxane (5.0 mL). This mixture was stirred at room temperature for 4 h and 20 concentrated under reduced re. The crude product was washed by ethyl acetate to 486 afford 337c (450 mg, 55%) as a pale yellow solid, which was used in the next step without further cation. MS-ESI: [M+H]+ 112.2 Example 337d 5-Bromo(1,2-dimethyl-1H-imidazolylamino) methylpyridin-2(1H)-one 337d 5 A 100-mL single-neck round-bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 337c (400 mg, 3.60 mmol), 3,5-dibromomethylpyridin- 2(1H)-one (960 mg, 3.60 mmol), XantPhos (240 mg, 0.40 mmol), tris(dibenzylideneacetone)dipalladium(0) (360 mg, 0.40 mmol), Cs2CO3 (4.69 g, 14.4 mmol), and 1,4-dioxane (20 mL). After three cycles of /nitrogen flush, the mixture was 10 heated at 90oC for 2.5 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica-gel column chromatography eluting with dichloromethane/methanol (30:1 to 20:1) to afford 337d as a pale yellow solid (350 mg, 33%). MS-ESI: [M+H]+ 297.1. e 337e [(1,2-Dimethyl-1H-imidazolyl)amino]methyloxo-1,6- 15 dihydropyridinyl}{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]-dodeca-2(6),7- dienyl}pyridinyl)methyl Acetate 337e A 50-mL single-neck round-bottomed flask equipped with a magnetic r and a reflux condenser was charged with 337d (20mg, 0.67 mmol), cetoxy)methyl]{4,4- dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic 20 acid 199e (270 mg, 0.67 mmol), Pd(dppf)Cl2 (42 mg, 0.050 mmol), sodium acetate (82 mg, 1.0 mmol), K3PO4 trihydrate (266 mg, 1.0 mmol), water (6 , and acetonitrile (6 mL).
After three cycles of vacuum/nitrogen flush, the e was heated at 95oC for 2 h. It was then filtered and the filtrate was evaporated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 25:1 dichloromethane/methanol to afford 25 337e (200 mg, 50%) as a brown solid. LCMS-ESI: [M+H]+ 570.3 Example 337 3-[4-[5-[(1,2-dimethylimidazolyl)amino]methyloxo pyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 337 A mixture of 337e (100 mg, 0.19 mmol) and lithium hydroxide (34 mg, 1.4 mmol) in 30 i-propanol /THF (1:1, 4 mL) and water (1 mL) was stirred at 40 oC for 0.5 h. The e was evaporated under reduced pressure and the residue was diluted with water (10 mL). It was then extracted with ethyl acetate (2 x 10 mL). The combined ethyl acetate extract was concentrated under reduced pressure and the residue was purified by reverse-phase prep- HPLC to afford 337 (35 mg, 40%) as a white solid. LCMS-ESI: [M+H]+ 528.3. 1H NMR 487 (500 MHz, CDCl3) δ 8.45 (d, J = 5.0 Hz, 1H), 7.36 (s, 1H), 7.28-7.27 (m, 2H), 7.25 (d, J = 5.0 Hz, 1H), 6.84 (s, 1H), 6.79 (s, 1H), 4.62-4.40 (m, 3H), 4.15-4.14 (m, 2H), 3.84-3.81 (m, 1H), 3.67 (s, 3H), 3.52 (s, 3H), 2.57-2.56 (m, 2H), 2.51 (s, 2H), 2.32 (s, 3H), 1.27 (s, 6H).
Example 338a 3-(2-Nitro-6,7-dihydropyrazolo[1,5-a]pyrazin-5(4H)- 5 yl)propanenitrile 338a N N N Br Br N N N O N N Pd/C, H2 N NH N O N EtOH, r.t., 1h N N NH2 Pd2(dba)3, xantphos, NO2 Cs2CO3, dioxane, N Br 100 °C, 5h 338a 338 N 338c b N N N NH AcO O N 199e N N Pd(dppf)Cl2 (0.05 eq), O N K3PO4 (2 eq), NaOAC 3H2O(2 eq), CH3CN, 90 oC, 1h 338d Following the procedure of Example 296d, and starting with 1-(2-bromoethyl) omethyl)nitro-1H-pyrazole 296d (268 mg, 1.00 mmol) and opropanenitrile (210 mg, 3.00 mmol) afforded 338a as a white solid (180 mg, 81%). MS-ESI: [M+H]+ 222.1 10 Example 338b 3-(2-Amino-6,7-dihydropyrazolo[1,5-a]pyrazin-5(4H)- yl)propanenitrile 338b Following the procedure of e 296e, and starting with 338a (180 mg, 0.81 mmol) afforded 338b as a yellow solid (120 mg, 77%). MS-ESI: [M+H]+ 192.2 Example 338c 3-(2-(5-Bromomethyloxo-1,2-dihydropyridinylamino)- 15 6,7-dihydropyrazolo[1,5-a]pyrazin-5(4H)-yl)propanenitrile 338c 488 Following the procedure of Example 309c, and starting with 338b (120 mg, 0.63 mmol) and 3,5-dibromomethylpyridin-2(1H)-one (169 mg, 0.63 mmol) afforded 338c as a yellow solid (150 mg, 63%). : [M+H]+ 377.2 Example 338d [4-(5-{[5-(2-Cyanoethyl)-4H,5H,6H,7H-pyrazolo[1,5- 5 a]pyrazinyl]amino}methyloxo-1,6-dihydropyridinyl){4,4-dimethyloxo- 1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl]methyl Acetate 338d Following the procedure of Example 309d, and starting with 338c (150 mg, 0.45 mmol) and {3-[(acetoxy)methyl]{4,4-dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- 2(6),7-dienyl}pyridinyl}boronic acid 199e (358 mg, 0.90 mmol) afforded 338d as a 10 yellow solid (150 mg, 52%). MS-ESI: [M+H]+ 650.3 Example 338 3-[2-[[5-[2-(7,7-dimethyloxo-1,2,6,8- ydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinyl)(hydroxymethyl)pyridyl] methyloxopyridyl]amino]-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazinyl]propanenitrile 338 15 Following the procedure of Example 309, and starting with 338e (150 mg, 0.23 mmol) afforded 338 as a white solid (55 mg, 40%). MS-ESI: [M+H]+ 608.3. 1H NMR (500 MHz, CDCl3) δ 8.49 (d, J = 5.0 Hz, 1H), 7.96 (d, J = 2.5 Hz, 1H), 7.71 (d, J = 2.0 Hz, 1H), 7.42 (s, 1H), 7.35 (d, J = 5.0 Hz, 1H), 6.85 (s, 1H), 5.74 (s, 1H), 5.05 (t, J = 6.5 Hz, 1H), 4.66-4.64 (m, 1H), 4.52-4.50 (m, 1H), 4.36-4.34 (m, 1H), 4.17-4.16 (m, 2H), 4.09-4.07 (m, 20 2H), 3.88-3.84 (m, 1H), 3.75 (s, 2H), 3.71 (s, 3H), 3.05-3.03 (m, 2H), 2.93-2.90 (m, 2H), 2.63-2.58 (m, 4H), 2.53 (s, 2H), 1.29 (s, 6H).
Example 339a tert-Butyl 4-(6-Nitropyridinyl)piperazinecarboxylate 339a To a solution of 5-bromonitropyridin (30.0 g, 148 mmol) in DMSO (1 L) were 25 added K2CO3 (40.0 g, 296 mmol) and tert-butyl zinecarboxylate (28.0 g, 148 mmol).
The mixture was stirred at 65 oC overnight. After cooling down, it was poured into water (2 L). The solid precipitated was collected and dried in vacuo. It was then further purified by -gel column chromatography eluting with 20:1 petroleum ethyl acetate and then with dichloromethane to afford 339a as a yellow solid (17.0 g, 37%). MS-ESI: [M+H]+ 309. 30 e 339b tert-Butyl 4-(6-Aminopyridinyl)piperazinecarboxylate 339b A 500-mL round-bottomed flask was purged with nitrogen and charged with 339a (3.1 g, 10 mmol), 10% palladium on carbon (50% wet, 1.0 g), and ethanol (100 mL). It was evacuated, charged with en gas via a balloon, and stirred at room temperature for 16 h. 489 The hydrogen was then evacuated and nitrogen was charged into the flask. The st was removed by filtration through a pad of CELITE® and the filtrate was concentrated under reduced pressure to afford 339b (2.7 g, 97%). MS-ESI: [M+H]+ 279 Example 339c tert-Butyl 4-(6-(5-Bromomethyloxo-1,2-dihydropyridin- 5 3-ylamino)pyridineyl)piperazinecarboxylate 339c A 100-mL single-neck bottomed flask equipped with a magnetic stirrer and a reflux condenser was charged with 339b (1.3 g, 4.7 mmol), 3,5-dibromomethylpyridin- 2(1H)-one (1.24 g, 4.7 mmol), cesium carbonate (3.8 g, 12 mmol), and 1,4-dioxane (50 mL).
After bubbling en through the resulting mixture for 30 minutes, XantPhos (272 mg, 10 0.47 mmol) and tris(dibenzylideneacetone)dipalladium(0) (430 mg, 0.47 mmol) were added.
The system was subjected to three cycles of argon/vacuum flush and heated at reflux for 3 h.
After this time the on was cooled to room temperature and filtered. The filtrate was partitioned between ethyl acetate (100 mL) and water (100 mL). The aqueous layer was separated and extracted with ethyl acetate (2 X 50 mL). The combined c layer was 15 washed with brine (50 mL) and dried over sodium sulfate. The drying agent was removed by filtration and the filtrate was trated under reduced pressure. The e was purified by silica-gel column chromatography eluting with 50:1 dichloromethane/methanol to afford 339c (1.3 g, 59%). MS-ESI: [M+H]+ 464.
Example 339d tert-Butyl 4-{6-[(5-{3-[(Acetoxy)methyl]{4,4-dimethyl 20 oxo-1,10-diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}methyloxo- 1,2-dihydropyridinyl)amino]pyridinyl}piperazinecarboxylate 339d 490 Boc BocN N N N N NH N NH O 199e AcO O N N Br PdCl2(dppf), N N K3PO4,NaOAc, MeCN, 95 °C, O N 1 h, H2O 339c 339d HN N N NH HCl/MeOH AcO O N N N O N 339e A 50-mL single-neck round-bottomed flask equipped with a magnetic r and a reflux condenser was charged with (2-{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}(4,4,5,5-tetramethyl-1,3,2-dioxaborolan- 5 2-yl)pyridinyl)methyl acetate 199e (287.4 mg, 0.60 mmol), 339c (145 mg, 0.30 mmol), Pd(dppf)Cl2 (24.5 mg, 0.030 mmol), K3PO4 (127.2 mg, 0.60 mmol), sodium acetate (49.2 mg, 0.60 mmol), water (0.50 mL), and itrile (10 mL). After three cycles of vacuum/nitrogen flush, the e was heated at 95 oC for 1 h. It was then cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure and the 10 resulting residue was purified by silica-gel column chromatography eluting with ethyl acetate to afford 339d as a yellow solid (140 mg, 61%). MS-ESI: [M+H]+ 737.3.
Example 339e (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- -dienyl}(1-methyloxo{[5-(piperazinyl)pyridinyl]amino}-1,6- dihydropyridinyl)pyridinyl)methyl Acetate 339e 15 A mixture of 339d (130 mg, 0.18 mmol) and HCl/methanol (4.0 mL) was stirred at room temperature for 4 h. It was then concentrated under reduced pressure to afford crude 339e (100 mg, 87%), which was used in the next step without further purification. MS-ESI: [M+H]+ 637.3. 491 Example 339 3-[3-(hydroxymethyl)[5-[[5-[4-(2-methoxyethyl)piperazinyl] pyridyl]amino]methyloxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 339 A mixture of 339e (100 mg, 0.18 mmol), 1-bromomethoxyethane (24.8 mg, 0.18 5 mmol), and K2CO3 (49.7 mg, 0.36 mmol) in acetonitrile (5.0 mL) in a sealed was stirred at 85 oC overnight. The mixture was cooled to room temperature and ed. The filtrate was concentrated under reduced pressure. To the residue was added water and the resulting mixture was extracted with dichloromethane three times. The combined organic layer was concentrated under reduced pressure and the resulting residue was purified by reverse-phase 10 prep-HPLC to afford 339 as a yellow solid (31.1 mg, 30%). MS-ESI: [M+H]+ 653.3. 1H NMR (500 MHz, DMSO) δ 8.61 (d, J = 2.0 Hz, 1H), 8.49 (d, J = 5.0 Hz, 1H), 8.40 (s, 1H), 7.84 (d, J = 3.0 Hz, 1H), 7.46 (d, J = 2.0 Hz, 1H), 7.38-7.34 (m, 2H), 7.23 (d, J = 9.5 Hz, 1H), 6.57 (s, 1H), .95 (m, 1H), 4.45-4.40 (m, 2H), 4.23-4.19 (m, 3H), 3.85-3.83 (m, 1H), 3.60 (s, 3H), 3.47-3.44 (m, 2H), 3.24 (s, 3H), 3.04-3.02 (m, 4H), 2.59-2.53 (m, overlap, 8H), 15 2.43 (s, 2H), 1.23 (s, 6H).
Example 340a (3S)-tert-Butyl yl(6-nitropyridinyl)piperazine carboxylate 340a Following the procedure of Example 323a, and starting with (3S)-tert-butyl 3- methylpiperazinecarboxylate (10.0 g, 50 mmol) and 5-bromonitropyridine (10.5 g, 50 20 mmol) afforded 340a as a yellow solid (8.05 g, 50%). MS-ESI: [M+H]+ 323 Example 340b (3S)-tert-Butyl 4-(6-Aminopyridinyl)methylpiperazine carboxylate 340b Following the ure of Example 323b, and starting with 340a(5.8 g, 18 mmol) afforded 340b as a brown solid (4.9 g, 93%). MS-ESI: [M+H]+ 293 25 Example 340c (3S)-tert-Butyl 4-(6-(5-Bromomethyloxo-1,2- dihydropyridinylamino) pyridineyl)methylpiperazinecarboxylate 340c ing the procedures of Example 323c, and starting with 340b (4.0 g, 13.7 mmol) and 3,5-dibromomethylpyridin-2(1H)-one (5.5 g, 20.6 mmol) afforded 340c as a yellow solid (5.4 g, 83%). : [M+H]+ 478 30 Example 340d (3S)Bromomethyl(5-(2-methylpiperazinyl)pyridin- ino)pyridine-2(1H)-one 340d Following the procedure of Example 271c, and starting with 340c (3.1 g, 6.5 mmol) afforded 340d as a yellow solid (2.3 g, 94%). MS-ESI: [M+H]+ 378. 492 Example 340e (S)Bromo(5-(4-(2-methoxyethyl)methylpiperazin yl)pyridinylamino)methylpyridin-2(1H)-one 340e O HN N N N O N NH Br N NH O O K2CO3, ACN, 80 oC N sealed tube, 12 h N Br Br 340d 340e O N N 199e N NH Pd(dppf)Cl2, AcO O N K3PO4, NaOAc, N N ACN, water, 100 oC, 1 h O N 340f A mixture of 340d (500 mg, 1.32 mmol), 1-bromomethoxyethane (239.1 mg, 1.72 5 mmol), K2CO3 (364 mg, 2.64 mmol), and acetonitrile (6 mL) in a sealed tube was heated at 80 oC for 12 h. Analysis of the reaction mixture by LCMS showed complete conversion to the desired product. It was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue was dissolved in water (20 mL) and ethyl acetate (30 mL). The water phase was extracted with ethyl acetate (3 X 30 mL). The 10 combined organic layer was washed with brine, dried over Na2SO4, concentrated under d pressure to afford crude 340e as a dark oil (600 mg), which was used in the next step without further purification. MS-ESI: [M+H]+ 436.1 Example 340f (2-{4,4-Dimethyloxo-1,10-diazatricyclo[6.4.0.02,6]dodeca- -dien yl}[5-({5-[(2S)(2-methoxyethyl)methylpiperazinyl]pyridin 15 yl}amino)methyloxo-1,6-dihydropyridinyl]pyridinyl)methyl Acetate 340f A 50-mL round ed flask equipped with a reflux condenser was charged with 340e (180 mg, 0.412 mmol), {3-[(acetoxy)methyl]{4,4-dimethyloxo-1,10- diazatricyclo[6.4.0.02,6]dodeca-2(6),7-dienyl}pyridinyl}boronic acid 199e (327.3 mg, 0.824 mmol), Pd(dppf)Cl2 (16.8 mg, 0.0206 mmol), K3PO4 (174.7 mg, 0.824 mmol), sodium 493 acetate (67.6 mg, 0.824 mmol), acetonitrile (10 mL), and water (3 drops). The system was subjected to three cycles of vacuum/nitrogen flush and heated at 100oC under N2 protection for 1 h. Analysis of the reaction mixture by LCMS showed complete conversion to the desired product. It was then cooled to room temperature and filtered. The filtrate was 5 concentrated under reduced pressure. The residue was purified by silica-gel column chromatography eluting with 20:1 dichloromethane/methanol to afford 340f as a yellow oil (190 mg, 65%). MS-ESI: [M+H]+ 709.4 Example 340 3-[3-(hydroxymethyl)[5-[[5-[(2S)(2-methoxyethyl)methylpiperazinyl ridyl]amino]methyloxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- 10 tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone 340 To a solution of 340f (170 mg, 0.24 mmol) in THF (6 mL), i-propanol (6 mL), and water (6 mL) was added lithium hydroxide (57.6 mg, 2.4 mmol). After stirring at room temperature for 1 h, The reaction mixture was concentrated under reduced pressure. The e was dissolved in romethane (20 mL) and water (10 mL). The water phase was 15 extracted with dichloromethane (3 x 20 mL). The combined organic layer was washed with brine, dried over Na2SO4, concentrated under reduced pressure, and purified by reverse-phase prep-HPLC to afford 340 (48.5 mg, 30%) as a white solid. MS-ESI: [M+H]+ 667.3. 1H NMR (500 MHz, DMSO-d6) δ 8.62 (d, J = 2.5 Hz, 1H), 8.49 (d, J = 5.0 Hz, 1H), 8.42 (s, 1H), 7.82 (d, J = 2.5 Hz, 1H), 7.47 (d, J = 2.5 Hz, 1H), 7.37-7.34 (m, 2H), 7.24 (d, J = 9.0 Hz, 1H), 20 6.56 (s, 1H), 4.97-4.95 (m, 1H), 4.47-4.41 (m, 2H), 4.25-4.19 (m, 3H), 3.85-3.83 (m, 1H), 3.63-3.62 (m, 1H), 3.61 (s, 3H), 3.47-3.45 (m, 2H), 3.25 (s, 3H), 3.06-3.04 (m, 1H), 2.93- 2.89 (m, 1H), 2.70-2.68 (m, 1H), 2.62-2.32 (m, overlap, 9H), 1.22 (s, 6H), 0.91 (d, J = 6.5 Hz, 3H).
Example 901 Biochemical Btk Assay 25 A generalized procedure for a standard biochemical Btk Kinase Assay that can be used to test Formula I compounds is as follows. A master mix minus Btk enzyme is prepared containing 1X Cell ing kinase buffer (25 mM Tris-HCl, pH 7.5, 5 mM betaglycerophosphate , 2 mM dithiothreitol, 0.1 mM Na3VO4, 10 mM MgCl2), 0.5 µM Promega PTK Biotinylated peptide substrate 2, and 0.01% BSA. A master mix plus Btk enzyme is 30 prepared ning 1X Cell Signaling kinase buffer, 0.5 µM PTK Biotinylated e substrate 2, 0.01% BSA, and 100 ng/well (0.06 mU/well) Btk enzyme. Btk enzyme is prepared as follows: full length human pe Btk sion number NM-000061) with a inal V5 and 6x His tag was subcloned into pFastBac vector for making baculovirus carrying this epitope-tagged Btk. Generation of virus is done based on Invitrogen's 494 ctions detailed in its published protocol "Bac-to-Bac Baculovirus Expression Systems" (Cat. Nos. 10359-016 and 10608-016). e 3 virus is used to infect Sf9 cells to overexpress the recombinant Btk protein. The Btk protein is then purified to homogeneity using Ni-NTA column. The purity of the final protein preparation is greater than 95% based 5 on the sensitive Sypro-Ruby staining. A solution of 200 µM ATP is prepared in water and adjusted to pH7.4 with 1N NaOH. A ty of 1.25 µL of compounds in 5%DMSO is transferred to a 96-well ½ area Costar polystyrene plate. nds are tested singly and with an 11-point dose-responsive curve (starting concentration is 10 µM; 1:2 dilution). A quantity of 18.75 µL of master mix minus enzyme (as a negative control) and master mix plus 10 enzyme is transferred to appropriate wells in 96-well ½ area costar polystyrene plate. 5 µL of 200 µM ATP is added to that mixture in the 96-well ½ area Costar polystyrene plate for final ATP concentration of 40 µM. The reaction is allowed to incubate for 1 hour at room temperature. The reaction is stopped with Perkin Elmer 1X detection buffer containing 30 mM EDTA, 20 nM SA-APC, and 1 nM PT66 Ab. The plate is read using time-resolved 15 fluorescence with a Perkin Elmer Envision using tion filter 330 nm, emission filter 665 nm, and 2nd emission filter 615 nm. IC50 values are subsequently ated. Alternatively, the Lanthascreen assay can be used to evaluate Btk activity through quantification of its phosphorylated peptide product. The FRET (Fluorescence Resonance Energy Transfer) that occurs between the fluorescein on the peptide product and the terbium on the detection 20 antibody decreases with the addition of inhibitors of Btk that reduce the phosphorylation of the peptide. In a final reaction volume of 25 uL, Btk (h) (0.1 ng/25 ul reaction) is incubated with 50 mM Hepes pH 7.5, 10 mM MgCl2, 2 mM MnCl2, 2 mM DTT, 0.2 mM NaVO4, 0.01% BSA, and 0.4 uM fluorescein poly-GAT. The on is initiated by the addition of ATP to 25 uM (Km of ATP). After incubation for 60 minutes at room temperature, the 25 reaction is stopped by the addition of a final tration of 2 nM Tb-PY20 detection dy in 60 mM EDTA for 30 minutes at room temperature. Detection is determined on a Perkin Elmer Envision with 340 nM excitation and emission at 495 nm and 520 nm.
Exemplary Btk inhibition IC50 values are in Tables 1, 2, and 3.
Example 902 Ramos Cell Btk Assay 30 Another lized procedure for a standard cellular Btk Kinase Assay that can be used to test Formula I nds is as follows. Ramos cells are incubated at a density of 0.5x107 cells/ml in the presence of test compound for 1 hr at 37 °C. Cells are then ated by incubating with 10 µg/ml anti-human IgM F(ab)2 for 5 minutes at 37 °C. Cells are 495 ed, lysed, and a protein assay is performed on the cleared lysate. Equal protein amounts of each sample are subject to SDS-PAGE and western blotting with either antiphosphoBtk (Tyr223) antibody (Cell Signaling Technology #3531; Epitomics, cat. #2207-1) or phosphoBtk(Tyr551) antibody (BD Transduction Labs #558034) to assess Btk 5 autophosphorylation or an anti-Btk antibody (BD Transduction Labs #611116) to control for total amounts of Btk in each .
Example 903 B-Cell Proliferation Assay A generalized ure for a standard cellular B-cell proliferation assay that can be used to test Formula I compounds is as follows. B-cells are purified from spleens of 8-16 10 week old Balb/c mice using a B-cell isolation kit (Miltenyi Biotech, Cat # 130862).
Testing compounds are diluted in 0.25% DMSO and incubated with 2.5 x 105 purified mouse splenic B-cells for 30 min prior to addition of 10µg/ml of an anti-mouse IgM dy (Southern Biotechnology Associates Cat # 1022-01) in a final volume of 100 µl. Following 24 hr incubation, 1 µCi 3H-thymidine is added and plates are incubated an additional 36 hr 15 prior to harvest using the manufacturer’s protocol for SPA[3H] thymidine uptake assay system (Amersham ences # RPNQ 0130). SPA-bead based fluorescence is counted in a microbeta counter (Wallace Triplex 1450, Perkin Elmer).
Example 904 T Cell Proliferation Assay A generalized procedure for a rd T cell proliferation assay that can be used to 20 test Formula I compounds is as follows. T cells are purified from spleens of 8-16 week old Balb/c mice using a Pan T cell isolation kit (Miltenyi Biotech, Cat # 130861). Testing nds are diluted in 0.25% DMSO and incubated with 2.5 x 105 purified mouse splenic T cells in a final volume of 100 µl in flat clear bottom plates precoated for 90 min at 37oC with 10 µg/ml each of anti-CD3 (BD # 553057) and anti-CD28 (BD # 553294) antibodies. 25 Following 24 hr tion, 1 µCi 3H-thymidine is added and plates incubated an additional 36 hr prior to harvest using the cturer’s protocol for SPA[3H] thymidine uptake assay system (Amersham Biosciences # RPNQ 0130). SPA-bead based fluorescence was counted in a microbeta r (Wallace x 1450, Perkin Elmer). e 905 CD86 Inhibition Assay 30 A generalized procedure for a standard assay for the inhibition of B cell activity that can be used to test Formula I nds is as follows. Total mouse splenocytes are purified from spleens of 8-16 week old Balb/c mice by red blood cell lysis (BD Pharmingen #555899).
Testing compounds are diluted to 0.5% DMSO and incubated with 1.25 x 106 splenocytes in a final volume of 200 µl in flat clear bottom plates (Falcon 353072) for 60 min at 37oC. Cells 496 are then stimulated with the addition of 15 µg/ml IgM (Jackson ImmunoResearch 6- 020), and incubated for 24 hr at 37oC, 5% CO2. Following the 24 hr incubation, cells are transferred to conical bottom clear 96-well plates and pelleted by fugation at 1200 x g x 5 min. Cells are preblocked by D32 (BD Pharmingen #553142), followed by triple 5 staining with CD19-FITC (BD Pharmingen #553785), CD86-PE (BD Pharmingen #553692), and 7AAD (BD Pharmingen #51-68981E). Cells are sorted on a BD libur and gated on the CD19+/7AAD- population. The levels of CD86 surface expression on the gated population is measured versus test compound concentration.
Example 906 B-ALL Cell Survival Assay 10 The following is a procedure for a standard B-ALL (acute lymphoblastic leukemia) cell al study using an XTT readout to measure the number of viable cells. This assay can be used to test a I compounds for their ability to inhibit the survival of B-ALL cells in culture. One human B-cell acute lymphoblastic leukemia line that can be used is SUP-B15, a human Pre-B-cell ALL line that is available from the ATCC. 15 SUP-B15 pre-B-ALL cells are plated in multiple 96-well microtiter plates in 100 µl of Iscove’s media + 20% FBS at a concentration of 5 x 105 cells/ml. Test compounds are then added with a final conc. of 0.4% DMSO. Cells are incubated at 37°C with 5% CO2 for up to 3 days. After 3 days cells are split 1:3 into fresh 96-well plates containing the test compound and allowed to grow up to an additional 3 days. After each 24h period, 50 ul of an XTT 20 solution is added to one of the replicate 96-well plates and absorbance readings are taken at 2, 4 and 20 hours following manufacturer’s directions. The reading taken with an OD for DMSO only treated cells within the linear range of the assay (0.5- 1.5) is then taken and the tage of viable cells in the compound treated wells are measured versus the DMSO only treated cells. 25 Example 907 CD69 Whole Blood Assay Human blood is obtained from healthy volunteers, with the ing restrictions: 1 week drug-free, non-smokers. Blood ximately 20 mls to test 8 compounds) is collected by venipuncture into Vacutainer® (Becton, Dickinson and Co.) tubes with sodium heparin. 30 Solutions of Formula I compounds at 10 mM in DMSO are d 1:10 in 100% DMSO, then are d by three-fold serial dilutions in 100% DMSO for a ten point doseresponse curve. The compounds are further diluted 1:10 in PBS and then an aliquot of 5.5 µl of each compound is added in duplicate to a 2 ml 96-well plate; 5.5 µl of 10% DMSO in PBS is added as control and no-stimulus wells. Human whole blood – HWB (100 µl) is added to 497 each well. After mixing the plates are incubated at 37 °C, 5% CO2, 100% humidity for 30 minutes. Goat F(ab’)2 anti-human IgM (10 µl of a 500 µg/ml on, 50 µg/ml final) is added to each well (except the mulus wells) with mixing and the plates are incubated for an additional 20 hours. At the end of the 20 hour incubation, samples are incubated with 5 fluorescent labeled antibodies for 30 minutes, at 37 °C, 5% CO2, 100% humidity. Include induced control, ned and single stains for compensation adjustments and initial voltage settings. s are then lysed with PharM Lyse™ (BD Biosciences Pharmingen) according to the manufacturer’s instructions. Samples are then transferred to a 96 well plate suitable to be run on the BD Biosciences HTS 96 well system on the LSRII machine. Data acquired and 10 Mean Fluorescence Intensity values were obtained using BD Biosciences DIVA Software.
Results are initially analyzed by FACS analysis software (Flow Jo). The inhibitory trations (IC50, IC70, IC90, etc.) for test compounds is defined as the concentration which decreases by, for example 50%, the percent positive of CD69 cells that are also CD20 positive stimulated by anti-IgM (average of 8 control wells, after subtraction of the average 15 of 8 wells for the no-stimulus background). The IC70 values are calculated by Prism version 5, using a nonlinear sion curve fit and are shown in Tables 1 and 2.
Example 908 in vitro Cell eration Assay Efficacy of Formula I compounds are measured by a cell proliferation assay employing the ing protocol (Mendoza et al (2002) Cancer Res. 62:5485-5488). The 20 CellTiter-Glo® Luminescent Cell Viability Assay, including reagents and protocol are commercially available (Promega Corp., Madison, WI, Technical in TB288). The assay assesses the ability of compounds to enter cells and inhibit cell proliferation. The assay principle is based on the determination of the number of viable cells t by quantitating the ATP present in a homogenous assay where addition of the iter Glo reagent results 25 in cell lysis and generation of a luminescent signal through the luciferase reaction. The luminescent signal is proportional to the amount of ATP present.
A panel of B-cell lymphoma cell lines (BJAB, 4, TMD8, OCI-Ly10, OCILy3 , CL2) are plated into 384-well plate in normal growth medium, and serially diluted BTK inhibitors or DMSO alone were added to each well. Cell viability is assessed 30 after 96 hour tion by CellTiter-Glo® (Promega). Data may be presented as Relative cell viability in BTK inhibitor-treated cells relative to DMSO-treated control cells. Data points are the mean of 4 replicates at each dose level. Error bars represent SD from the mean. 498 Procedure: Day 1 – Seed Cell Plates (384-well black, clear , microclear, TC plates with lid from Falcon #353962), Harvest cells, Seed cells at 1000 cells per 54µl per well into 384 well Cell Plates for 3 days assay. Cell Culture Medium: RPMI or DMEM high glucose, 10% Fetal Bovine Serum, 2mM L-Glutamine, P/S. Incubate O/N at 37 ºC, 5% CO2. 5 Day 2 – Add Drug to Cells, Compound Dilution, DMSO Plates (serial 1:2 for 9 points), Add 20 µl compounds at 10 mM in the 2nd column of 96 well plate. Perform serial 1:2 across the plate (10µl + 20µl 100% DMSO) for a total of 9 points using Precision. Media Plates 96-well l bottom polypropylene plates from Nunc (cat.# 249946) (1:50 dilution) Add 147µl of Media into all wells. er 3µl of DMSO + compound from each well in 10 the DMSO Plate to each corresponding well on Media Plate using Rapidplate.
Drug Addition to Cells, Cell Plate (1:10 dilution), Add 6µl of media + compound directly to cells (54µl of media on the cells already). Incubate 3 days at 37 C, 5% CO2 in an incubator that will not be opened often.
Day 5 – Develop Plates, Thaw Cell Titer Glo Buffer at room temperature. Remove 15 Cell Plates from 37 °C and equilibrate to room temperature. for about 30 s. Add Cell Titer Glo Buffer to Cell Titer Glo Substrate (bottle to bottle). Add 30 µl Cell Titer Glo Reagent (Promega cat.# G7572) to each well of cells. Place on plate shaker for about 30 minutes. Read luminescence on Analyst HT Plate Reader (half second per well).
Cell viability assays and combination assays: Cells were seeded at 1000-2000 20 cells/well in 384-well plates for 16 h. On day two, nine serial 1:2 nd dilutions are made in DMSO in a 96 well plate. The nds are further diluted into growth media using a Rapidplate robot (Zymark Corp., Hopkinton, MA). The diluted compounds are then added to quadruplicate wells in 384-well cell plates and incubated at 37 ºC and 5% CO2.
After 4 days, relative s of viable cells are measured by scence using Cell-Titer 25 Glo (Promega) according to the manufacturer’s instructions and read on a Wallac Multilabel Reader (PerkinElmer, Foster City). EC50 values are calculated using Prism® 4.0 re (GraphPad, San Diego). Formula I compounds and chemotherapeutic agents are added simultaneously or separated by 4 hours (one before the other) in all assays.
An additional exemplary in vitro cell proliferation assay includes the following steps: 30 1. An aliquot of 100 µl of cell e containing about 104 cells in medium is deposited in each well of a 384-well, opaque-walled plate. 2. Control wells are prepared containing medium and without cells. 3. The compound is added to the mental wells and incubated for 3-5 days. 4. The plates are equilibrated to room temperature for approximately 30 minutes. 499 5. A volume of CellTiter-Glo Reagent equal to the volume of cell culture medium present in each well is added. 6. The contents are mixed for 2 minutes on an orbital shaker to induce cell lysis. 7. The plate is incubated at room temperature for 10 minutes to ize the 5 luminescence signal. 8. Luminescence is recorded and reported in graphs as RLU = relative luminescence units.
Although the ing invention has been described in some detail by way of 10 illustration and example for purposes of clarity of understanding, the ptions and examples should not be construed as limiting the scope of the invention. Accordingly, all suitable modifications and equivalents may be considered to fall within the scope of the invention as defined by the claims that . The disclosures of all patent and scientific literature cited herein are expressly incorporated in their entirety by reference. 15

Claims (44)

We Claim:
1. A compound selected from Formula I: or stereoisomers, tautomers, or pharmaceutically acceptable salts thereof, wherein: 5 X1 is CR1 or N; X2 is CR2 or N; X3 is CR3 or N; where one or two of X1, X2, and X3 are N; R1, R2 and R3 are independently selected from H, F, Cl, −NH2, −NHCH3, )2, − 10 OH, −OCH3, −OCH2CH3, −OCH2CH2OH, and C1−C3 alkyl; R4 is selected from H, F, Cl, CN, −CH2OH, −CH(CH3)OH, )2OH, − CH(CF3)OH, −CH2F, −CHF2, −CH2CHF2, −CF3, −C(O)NH2, −C(O)NHCH3, −C(O)N(CH3)2, −NH2, −NHCH3, −N(CH3)2, −NHC(O)CH3, −OH, −OCH3, −OCH2CH3, −OCH2CH2OH, cyclopropyl, cyclopropylmethyl, 1-hydroxycyclopropyl, imidazolyl, pyrazolyl, 3-hydroxy- 15 oxetanyl, yl, and azetidinyl; R5 is optionally tuted C6−C20 aryl, C3−C12 carbocyclyl, C2−C20 heterocyclyl, C1−C20 heteroaryl, −(C6−C20 aryl)−(C2−C20 cyclyl), −(C1−C20 heteroaryl)−(C2−C20 heterocyclyl), −(C1−C20 heteroaryl)−(C2−C20 heterocyclyl)−(C2−C20 heterocyclyl), −(C1−C20 heteroaryl)−(C2−C20 heterocyclyl)−(C1−C6 , −(C1−C20 heteroaryl)−(C1−C6 alkyl), −(C2− 20 C20 heterocyclyl)−(C1−C6 alkyl), −(C2−C20 heterocyclyl)−(C3−C12 carbocyclyl), −(C1−C20 heteroaryl)−(C3−C12 carbocyclyl), or −(C1−C20 heteroaryl)−C(=O)−(C2−C20 heterocyclyl); R6 is H, −CH3, −CH2CH3, −CH2CH2OH, −CHF2, −NH2, or −OH; R7 is selected from the structures: N N N N N N N N N N N N F O O F O N N N N N F O O F O N N N N N N N N N N O F O O N N N N S N F O O O O where the wavy line indicates the site of attachment; and Y1 and Y2 are independently selected from CH and N, where Y1 and Y2 are not each where alkyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl are optionally substituted 5 with one or more groups independently selected from F, Cl, Br, I, −CN, −CH3, −CH2CH3, − CH(CH3)2, −CH2CH(CH3)2, −CH2OH, H3, −CH2CH2OH, −C(CH3)2OH, − CH(OH)CH(CH3)2, −C(CH3)2CH2OH, −CH2CH2SO2CH3, −CH2OP(O)(OH)2, −CH2F, −CHF2, −CF3, 3, −CH2CHF2, −CH(CH3)CN, −C(CH3)2CN, −CH2CN, −CO2H, −COCH3, − CO2CH3, −CO2C(CH3)3, −COCH(OH)CH3, −CONH2, −CONHCH3, −CON(CH3)2, − 10 C(CH3)2CONH2, −NH2, −NHCH3, −N(CH3)2, −NHCOCH3, −N(CH3)COCH3, −NHS(O)2CH3, −N(CH3)C(CH3)2CONH2, −N(CH3)CH2CH2S(O)2CH3, −NO2, =O, −OH, −OCH3, − OCH2CH3, −OCH2CH2OCH3, −OCH2CH2OH, −OCH2CH2N(CH3)2, −OP(O)(OH)2, − S(O)2N(CH3)2, −SCH3, CH3, −S(O)3H, cyclopropyl, oxetanyl, inyl, 1- methylazetidinyl)oxy, N-methyl-N-oxetanylamino, azetidinylmethyl, and 15 morpholino.
2. The compound of claim 1 wherein X1 is N.
3. The compound of claim 1 wherein X2 is N.
4. The compound of claim 1 wherein X3 is N.
5. The compound of claim 1 wherein X1 and X3 are N, X1 and X2 are N, or X2 20 and X3 are N.
6. The compound of claim 1 wherein R5 is optionally substituted C1−C20 aryl selected from pyrazolyl, pyridinyl, pyrimidinyl, 5-methyl-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrazinyl, 5-acetyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin yl, 6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazinyl, and 1-methyl(5-(4-methylpiperazin 25 yl)pyridinyl.
7. The compound of claim 1 n R5 is −(C1−C20 heteroaryl)−(C2−C20 heterocyclyl) where heteroaryl is optionally substituted pyridinyl and heterocyclyl is ally substituted piperazinyl.
8. The compound of claim 1 wherein R5 is phenyl, optionally substituted with 30 one or more groups selected from F, Cl, −CH3, −S(O)2CH3, cyclopropyl, azetidinyl, oxetanyl, and lino.
9. The compound of claim 1 n R5 is selected from the structures: 504 504 O O N N N S N O N N O H H N N N O S O O O N N N N N S O S O S where the wavy line indicates the site of attachment.
10. The compound of claim 1 wherein R5 is: where R8 is ed from H, −CH3, −CH2OCH3, −CH2CH3, −CH(CH3)2, − CH2CH2OH, −CH2CH2OCH3, −CH2F, −CHF2, −CF3, −CH2CF3, −CH2CHF2, −CH(CH3)CN, −C(CH3)2CN, −CH2CN, −C(O)CH3, −C(O)CH2CH3, H(CH3)2, −NH2, , − 2, −OH, −OCH3, −OCH2CH3, −OCH2CH2OH, ropyl, and oxetanyl.
11. The compound of claim 1 wherein R6 is CH3.
12. The compound of claim 1 wherein Y1 is CH and Y2 is N. 5
13. The compound of claim 1 wherein Y1 is N and Y2 is CH.
14. The compound of claim 1 wherein Y1 and Y2 are each CH.
15. The compound of claim 1 wherein Y1 and Y2 are each CH, and R6 is CH3.
16. The compound of claim 1 selected from Table 1.
17. The nd of claim 1 selected from Table 2. 10
18. The compound of claim 1, which is 2-{4-Hydroxymethyl-1'-methyl-5'-[5-(4- oxetanyl-piperazinyl)-pyridinylamino]-6'-oxo-1',6'-dihydro-[3,3']bipyridinylyl}- 3,4,6,7,8,9-hexahydro-2H-pyrazino[1,2-a]indolone or a pharmaceutically acceptable salt thereof.
19. The compound of claim 1, which is 6-{3'-Hydroxymethylmethyl[5-(4- 15 oxetanyl-piperazinyl)-pyridinylamino]oxo-1,6-dihydro-[3,4']bipyridinyl-2'-yl}- 2,2-dimethyl-2,3,5,6-tetrahydro-1H,4Hthiaaza-cyclopenta[a]indenone or a pharmaceutically acceptable salt thereof.
20. The compound of claim 1, which is 2-{4-Hydroxymethyl-1'-methyl-5'-[5-(4- oxetanyl-piperazinyl)-pyridinylamino]-6'-oxo-1',6'-dihydro-[3,3']bipyridinylyl}- 20 6,7,8,9-tetrahydro-2H-pyrazino[1,2-a]indolone or a pharmaceutically acceptable salt thereof.
21. The compound of claim 1, which is 2-{3'-Hydroxymethylmethyl[5-((S)- 2-methyloxetanyl-piperazinyl)-pyridinylamino]oxo-1,6-dihydro- [3,4']bipyridinyl-2'-yl}-7,7-dimethyl-3,4,7,8-tetrahydro-2H,6H-cyclopenta[4,5]pyrrolo[1,2- 25 a]pyrazinone or a pharmaceutically acceptable salt thereof.
22. The compound of claim 1, which is 2-{3-Hydroxymethyl[1-methyl(5- methyl-4,5,6,7-tetrahydro-pyrazolo[1,5-a]pyrazinylamino)oxo-1,6-dihydro-pyridazin yl]-pyridinyl}-3,4,6,7,8,9-hexahydro-2H-pyrazino[1,2-a]indolone or a ceutically acceptable salt thereof. 30
23. The compound of claim 1, which is 2-[4-Hydroxymethyl-1'-methyl-5'-(5- methyl-4,5,6,7-tetrahydro-pyrazolo[1,5-a]pyrazinylamino)-6'-oxo-1',6'-dihydro- [3,3']bipyridinylyl]-3,4,6,7,8,9-hexahydro-2H-pyrazino[1,2-a]indolone or a pharmaceutically acceptable salt thereof.
24. The compound of claim 1, which is hydroxymethyl)[1-methyl[(2- methylpyrimidinyl)amino]oxopyridyl]pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone or a pharmaceutically able salt 5
25. The compound of claim 1, which is 2-[3-(hydroxymethyl)[5-(1H- imidazo[4,5-b]pyridinylamino)methyloxopyridyl]pyridyl]-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indolone or a pharmaceutically acceptable salt thereof.
26. The compound of claim 1, which is 3-[4-[5-[(2-ethylpyrimidinyl)amino] methyloxopyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8- 10 tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone or a pharmaceutically acceptable salt thereof.
27. The compound of claim 1, which is 2-[3-(hydroxymethyl)[1-methyl[[5- (1-methylpiperidyl)pyridyl]amino]oxopyridyl]pyridyl]-3,4,6,7,8,9- hexahydropyrazino[1,2-a]indolone or a pharmaceutically acceptable salt thereof. 15
28. The nd of claim 1, which is 5-[(5-acetyl-6,7-dihydro-4H- thiazolo[5,4-c]pyridinyl)amino]methyloxopyridyl](hydroxymethyl)pyridyl]- 10-fluoro-3,4,6,7,8,9-hexahydropyrazino[1,2-a]indolone or a pharmaceutically acceptable salt thereof.
29. The compound of claim 1, which is 10-fluoro[3-(hydroxymethyl)[1- 20 methyl[[5-[(2S)methyl(oxetanyl)piperazinyl]pyridyl]amino]oxo pyridyl]pyridyl]-3,4,6,7,8,9-hexahydropyrido[3,4-b]indolizinone or a pharmaceutically acceptable salt thereof.
30. The compound of claim 1, which is 3-[3-(hydroxymethyl)[5-[[5-[(2S) methyl(oxetanyl)piperazinyl]pyridyl]amino]oxo-1H-pyridazinyl] 25 pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone or a pharmaceutically acceptable salt thereof.
31. The compound of claim 1, which is 3-[4-[5-[(5-fluoropyridyl)amino] methyloxopyridyl](hydroxymethyl)pyridyl]-7,7-dimethyl-1,2,6,8- tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazinone or a pharmaceutically acceptable salt 30 thereof.
32. The compound of claim 1, which is 3-[3-(hydroxymethyl)[5-[[5-(3- methoxypropyl)-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazinyl]amino]methyloxo pyridyl]pyridyl]-7,7-dimethyl-1,2,6,8-tetrahydrocyclopenta[3,4]pyrrolo[3,5-b]pyrazin one or a pharmaceutically acceptable salt thereof.
33. A pharmaceutical composition comprised of a compound of any one of claims 1 to 32 and a pharmaceutically acceptable carrier, glidant, diluent, or excipient.
34. The pharmaceutical ition according to claim 33, further comprising a therapeutic agent. 5
35. A process for making a pharmaceutical composition which comprises combining a nd of any one of claims 1 to 32 with a pharmaceutically acceptable carrier.
36. Use of a pharmaceutical composition of claim 33 in the manufacture of a medicament for the treatment of immune disorders, cancer, cardiovascular disease, viral 10 ion, inflammation, metabolism/endocrine function disorders and ogical ers, and wherein the ment mediates the Bruton’s tyrosine kinase.
37. The use of claim 36 wherein the disease or disorder is an immune disorder.
38. The use of claim 37 wherein the immune disorder is rheumatoid arthritis.
39. The use of claim 36 wherein the disease or disorder is systemic and local 15 inflammation, arthritis, inflammation related to immune suppression, organ transplant rejection, allergies, ulcerative colitis, Crohn’s disease, dermatitis, asthma, systemic lupus erythematosus, Sjögren’s Syndrome, le sclerosis, scleroderma/systemic sclerosis, idiopathic thrombocytopenic purpura (ITP), anti-neutrophil cytoplasmic antibodies (ANCA) itis, chronic obstructive ary disease (COPD), psoriasis. 20
40. The use of claim 36 wherein the disease or disorder is cancer selected from breast, ovary, cervix, prostate, testis, genitourinary tract, esophagus, , glioblastoma, neuroblastoma, stomach, skin, keratoacanthoma, lung, epidermoid carcinoma, large cell carcinoma, non-small cell lung carcinoma (NSCLC), small cell carcinoma, lung arcinoma, bone, colon, adenoma, pancreas, arcinoma, thyroid, follicular 25 carcinoma, undifferentiated oma, papillary carcinoma, ma, melanoma, sarcoma, bladder carcinoma, liver carcinoma and biliary passages, kidney carcinoma, pancreatic, myeloid disorders, lymphoma, hairy cells, buccal cavity, haryngeal, pharynx, lip, tongue, mouth, small intestine, colon-rectum, large intestine, rectum, brain and central nervous system, Hodgkin’s, ia, bronchus, thyroid, liver and intrahepatic bile duct, 30 hepatocellular, gastric, glioma/glioblastoma, endometrial, melanoma, kidney and renal pelvis, urinary bladder, uterine corpus, uterine cervix, multiple myeloma, acute myelogenous leukemia, chronic myelogenous leukemia, lymphocytic leukemia, chronic lymphoid leukemia (CLL), myeloid leukemia, oral cavity and x, non-Hodgkin lymphoma, melanoma, and villous colon adenoma.
41. The use of claim 36 further comprising use of an additional therapeutic agent selected from an anti-inflammatory agent, an immunomodulatory agent, chemotherapeutic agent, an apoptosis-enhancer, a neurotropic factor, an agent for treating cardiovascular disease, an agent for treating liver disease, an anti-viral agent, an agent for treating blood 5 disorders, an agent for treating diabetes, and an agent for treating immunodeficiency disorders.
42. A kit for treating a condition mediated by Bruton’s ne kinase, comprising: a) a pharmaceutical composition of claim 33; and 10 b) instructions for use.
43. The pharmaceutical ition of claim 33 for use as a medicament in ng a disease or disorder selected from immune disorders, cancer, cardiovascular disease, viral infection, inflammation, metabolism/endocrine function disorders and neurological disorders, and ed by Bruton’s tyrosine kinase. 15
44. A pharmaceutical composition according to claim 33 ntially as herein described with reference to any example thereof.
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