EP1165575A1 - Phosphat-enthaltende makrozyclische immunomodulatoren - Google Patents

Phosphat-enthaltende makrozyclische immunomodulatoren

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Publication number
EP1165575A1
EP1165575A1 EP00918274A EP00918274A EP1165575A1 EP 1165575 A1 EP1165575 A1 EP 1165575A1 EP 00918274 A EP00918274 A EP 00918274A EP 00918274 A EP00918274 A EP 00918274A EP 1165575 A1 EP1165575 A1 EP 1165575A1
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EP
European Patent Office
Prior art keywords
minor
group
ethyl
dioxa
dimethoxy
Prior art date
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EP00918274A
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English (en)
French (fr)
Inventor
Megumi Kawai
Indrani W. Gunawardana
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Abbott Laboratories
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Abbott Laboratories
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Publication of EP1165575A1 publication Critical patent/EP1165575A1/de
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6561—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/10—Anti-acne agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/14—Drugs for dermatological disorders for baldness or alopecia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection

Definitions

  • the present invention relates to novel chemical compounds having immunomodulatory activity, and in particular to macrolide immunosuppressants. More particularly, the invention relates to semisynthetic analogs of ascomycin and FK-506. means for their preparation, pharmaceutical compositions containing such compounds and methods of treatment employing the same.
  • cyclosporine (cyclosporin A) has found wide use since its introduction in the fields of organ transplantation and immunomodulation, and has brought about a significant increase in the success rate for transplantation procedures. Unsatisfactory side-effects associated with cyclosporine, however, such as nephrotoxicity, have led to a continued search for irnmunosuppressant compounds having improved efficacy and safety.
  • FR-900520 (lb, Figure 1)
  • FR-900523 (lc.
  • Figure 1 which differ from FK-506 in their alkyl substituent at C-17 ( Figure 1), have been isolated from S. hygroscopicus yakushimnaensis.
  • Figure 1 Another analog, FR-900525 (Id, Figure 1), produced by S. tsukubaensis, differs from FK-506 by the replacement of the piperdine moiety with a pyrrolidine moiety.
  • FR-900520 also known as ascomycin
  • ascomycin has been previously disclosed by Arai et al. in U.S. Patent No. 3,244,592, issued April 5, 1966, where the compound is described as an antifungal agent.
  • Monaghan, R.L., et al. on the other hand, described the use of ascomycin as an irnmunosuppressant in European Patent Application No. 323865, published July 12, 1989.
  • FK-506 Although the immunosuppressive activity of FK-506 has been clinically confirmed, toxicity in mammals has limited its utility. The activity of FK-506 has, however, prompted efforts to discover novel analogs of FK-type compounds which possess superior properties. These efforts include the isolation of new fermentation products, the microbial transformation of existing chemical entities, the chemical modification of these macrocycles, and the synthesis of hybrid species derived from smaller synthetic fragments.
  • FK-type compounds include C-17-epi derivatives of FK- 506; a 3'-demethylated derivative of FK-506; 3'-oxo-FK-506 ; compounds derived from FK-506, FR-900523 and FR-900525 which are characterized by the introduction of hydroxy protecting groups; formation of a double bond by elimination of water between carbons 14 and 15 ; oxidation of the hydroxy group at carbon 14 to the ketone, and reduction of the allyl side-chain at carbon 17 via hydrogenation ( Figure 1).
  • Other published derivatives include those derived from FK-506 and FR-900520 where the lactone ring is contracted to give a macrocyclic ring containing two fewer carbons.
  • FK-type compounds have been published, such as the microbial demethylation of FR-900520 to form the bis-demethylated 3',25- dihydroxy ring-rearranged derivative of FR-900520; the microbial monodemethylation at carbon 25 of FK-506 and FR-900520; and the microbial monodemethylation of FR- 900520 at C-3' ( Figure 1), as well as a number of other macrocyclic microbial transformation products. Numerous chemical modifications of the FK-type compounds have been attempted.
  • one object of the present invention is to provide novel semisynthetic macrolides which possess the desired immunomodulatory activity but which may be found to minimize unwanted side effects. More particularly, the present invention provides novel semisynthetic macrolides which bear a phosphate moiety at C-4' ( Figure 1).
  • Another object of the present invention is to provide synthetic processes for the preparation of such compounds from starting materials obtained by fermentation, as well as chemical intermediates useful in such synthetic processes.
  • a further object of the present invention is to provide pharmaceutical compositions containing, as an active ingredient, one of the above compounds.
  • Yet another object of the invention is to provide a method of treating a variety of disease states, including post- transplant tissue rejection and autoimmune dysfunction.
  • R 2 and R 3 are independently selected from the group consisting of
  • R 7 is selected from the group consisting of a) hydrogen and b) hydroxy protecting group, or R 2 and R 3 taken together are selected from the group consisting of
  • R 4 is selected from the group consisting of (1) hydrogen and
  • R 7 is previously defined; R 3 and R 6 are independently selected from the group consisting of (1) hydrogen and
  • Z is selected from the group consisting of O and S;
  • A is selected from the group consisting of
  • R 82 and R 83 are independently selected from the group consisting of hydrogen, alkylcarbonyl, and formyl, provided that at least one of R 82 and R 8 ' is other than hydrogen;
  • R 9 is selected from the group consisting of
  • compositions which comprise a compound of formula I in combination with a pharmaceutically acceptable carrier alone or in combination with another compound of formula I.
  • a method for treating or preventing autoimmune diseases in a mammal comprising administering to the mammal a therapeutically effective amount of a compound of formula I.
  • Compounds of this invention include, but are not limited to,
  • Stereoisomers can exist as stereoisomers, wherein asymmetric or chiral centers are present. Stereoisomers are designated “R” or “ S,” depending on the configuration of substituents around the chiral carbon atom.
  • R and S used herein are configurations as defined in IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, Pure Appl. Chem., (1976), 45: 13-30, hereby incorporated by reference.
  • Stereoisomers include enantiomers, diastereomers, and mixtures of enantiomers or diastereomers.
  • Individual stereoisomers of compounds of the present invention may be prepared synthetically from commercially available starting materials which contain asymmetric or chiral centers or by preparation of racemic mixtures followed by resolution well-known to those of ordinary skill in the art.
  • alkenyl refers to a straight or branched chain hydrocarbon containing from 2 to 6 carbons and containing at least one carbon-carbon double bond formed by the removal of two hydrogens.
  • Representative examples of “alkenyl” include, but are not limited to, ethenyl, 2-propenyl, 2-methyl-2-propenyl, 3- butenyl, 4-pentenyl, 5-hexenyl, and the like.
  • alkoxy refers to an alkyl group, as defined herein, appended to the parent molecular moiety through an oxy group, as defined herein.
  • alkoxy include, but are not limited to. methoxy, ethoxy, propoxy, 2-propoxy, butoxy, tert-butoxy, pentyloxy, hexyloxy and the like.
  • alkoxycarbonyl refers to an alkoxy group, as defined herein, appended to the parent molecular moiety through a carbonyl group, as defined herein.
  • Representative examples of alkoxycarbonyl include, but are not limited to, methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbon l, and the like.
  • alkyl refers to a straight or branched chain hydrocarbon containing from l-to-6 carbon atoms.
  • Representative examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, n-hexyl, and the like.
  • alkylcarbonyl refers to an alkyl group, as defined herein, appended to the parent molecular moiety through a carbonyl group, as defined herein.
  • Representative examples of alkylcarbonyl include, but are not limited to, acetyl, 1- oxopropyl, 2,2-dimethyl-l-oxopropyl, 1-oxobutyl, 1-oxopentyl, and the like.
  • alkylcarbonyloxy refers to an alkylcarbonyl group, as defined herein, appended to the parent molecular moiety through an oxy group, as defined herein.
  • Representative examples of alkylcarbonyloxy include, but are not limited to, acetyloxy, ethylcarbonyloxy, tert-butylcarbonyloxy, and the like.
  • alkylene denotes a divalent group derived from a straight or branched chain hydrocarbon of from 1 to 6 carbon atoms.
  • Representative examples of alkylene include, but are not limited to. -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -. -CH 2 CH 2 CH 2 CH 2 -, -CH 2 CH(CH,)CH 2 -, and the like.
  • alkylthio refers to an alkyl group, as defined herein, appended to the parent molecular moiety through a thio group, as defined herein.
  • Representative examples of alkylthio include, but are not limited, methylthio, ethylthio, tert-butylthio, hexylthio, and the like.
  • alkynyl refers to a straight or branched chain hydrocarbon group containing from 2 to 6 carbon atoms and containing at least one carbon-carbon triple bond.
  • Representative examples of alkynyl include, but are not limited, to acetylenyl, 1-propynyl, 2-propynyl, 3-butynyl, 2-p ⁇ ntynyl, 1-butynyl and the like.
  • -C(O)NR 90 R 91 refers to a -C(O)NR 90 R 91 group, wherein R 90 and R 91 are independently selected from the group consisting hydrogen, alkyl, aryl, and arylalkyl, as defined herein.
  • Representative examples of -C(O)NR 90 R 91 include, but are not limited to, aminocarbonyl, dimethylaminocarbonyl, methylaminocarbonyl, diethyaminocarbonyl, benzylaminocarbonyl, and the like.
  • amino refers to a -NR 92 R 9 ' group, wherein R 92 and R 93 are independently selected from the group consisting hydrogen, alkoxycarbonyl, alkyl, alkylcarbonyl, aryl, arylalkoxycarbonyl, arylalkyl, and formyl, as defined herein.
  • Representative examples of -NR 92 R 93 include, but are not limited to, acetylamino, benzyloxycarbonylamino, formylamino, ethoxycarbonylamino, acetylmethylamino, and the like.
  • aryl refers to a phenyl group.
  • aryl groups of this invention can be substituted with 1, 2, or 3 substituents independently selected from alkoxy, alkoxycarbonyl. alkyl, alkylcarbonyl, alkylcarbonyloxy, alkylthio, amido, amino, carboxy. cyano. ethylenedioxy, formyl, halogen, haloalkyl, hydroxy, methylenedioxy, and nitro.
  • arylalkoxy refers to an aryl group, as defined herein, appended to the parent molecular moiety through an alkoxy group, as defined herein.
  • Representative examples of arylalkoxy include, but are not limited to, 2-phenylethoxy, 3- phenylpropoxy, 5-phenylpentyloxy, and the like.
  • arylalkoxycarbonyl refers to an arylalkoxy group, as defined herein, appended to the parent molecular moiety through a carbonyl group, as defined herein.
  • arylalkoxycarbonyl include, but are not limited to, benzyloxycarbonyl, 2-phenylethoxycarbonyl, and the like.
  • arylalkyl refers to an aryl group, as defined herein, appended to the parent molecular moiety through an alkyl group, as defined herein.
  • Representative examples of arylalkyl include, but are not limited to, benzyl, 2-phenylethyl, 3-phenylpropyl, and the like.
  • carbonyl refers to a -C(O)- group.
  • carboxy refers to a -CO 2 H group.
  • cyano refers to a -CN group.
  • cyanoalkyl refers to a cyano group, as defined herein, appended to the parent molecular moiety through an alkyl group, as defined herein.
  • Representative examples of cyanoalkyl include, but are not limited to, 2-cyanoethyl, 3-cyanopropyl, and the like.
  • cycloalkyl refers to a saturated cyclic hydrocarbon group containing from 3 to 8 carbons.
  • Representative examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like.
  • the cycloalkyl groups of this invention can be substituted with 1, 2,or 3 substituents independently selected from alkoxy, alkyl, halogen, haloalkyl, and -OR'.
  • cycloalkylalkyl refers to cycloalkyl group, as defined herein, appended to the parent molecular moiety through an alkyl group, as defined herein.
  • Representative examples of cycloalkylalkyl include, but are not limited to, cyclopropylmethyl, 2-cyclobutylethyl, cyclopentylmethyl, cyclohexylmethyl,
  • ethyl refers to a -CH 2 CH 3 group.
  • halo refers to -Cl, -Br, -I or -F.
  • haloalkyl refers to at least one halogen, as defined herein, appended to the parent molecular moiety through an alkyl group, as defined herein. Representative examples of haloalkyl include, but are not limited to, chloromethyl, 2-fluoroethyl, trifluoromethyl, pentafluoroethyl, 2-chloro-3-fluoropentyl, and the like.
  • heterocycle refers to a monocyclic, bicyclic, or tricyclic ring system.
  • Monocyclic ring systems are exemplified by any 3- or 4- membered ring containing a heteroatom independently selected from oxygen, nitrogen and sulfur; or a 5-, 6- or 7-membered ring containing one, two or three heteroatoms wherein the heteroatoms are independently selected from nitrogen, oxygen and sulfur.
  • the 5- membered ring has from 0-2 double bonds and the 6- and 7-membered ring have from 0-3 double bonds.
  • monocyclic ring systems include, but are not limited to, azetidine, azepine, aziridine, diazepine, 1,3-dioxolane, dioxane, dithiane, furan, imidazole, imidazoline, imidazolidine, isothiazole, isothiazoline, isothiazolidine, isoxazole, isoxazoline, isoxazolidine, morpholine, oxadiazole, oxadiazoline, oxadiazolidine, oxazole, oxazoline, oxazolidine, piperazine, piperidine, pyran, pyrazine, pyrazole, pyrazoline, pyrazolidine, pyridine, pyrimidine, pyridazine, pyrrole, pyrroline, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, tetrazine,
  • Bicyclic ring systems are exemplified by any of the above monocyclic ring systems fused to an aryl group as defined herein, a cycloalkyl group as defined herein, or another monocyclic ring system.
  • Representative examples of bicyclic ring systems include but are not limited to, for example, benzimidazole, benzthiazole, benzothiophene, benzoxazole. benzofuran, benzopyran, benzothiopyran, benzodioxine, 1,3-benzodioxole, cinnoline.
  • Tricyclic rings systems are exemplified by any of the above bicyclic ring systems fused to an aryl group as defined herein, a cycloalkyl group as defined herein, or a monocyclic ring system.
  • tricyclic ring systems include, but are not limited to, acridine, carbazole, carboline. dibenzofuran, dibenzothiophene, naphthofuran, naphthothiophene, oxanthrene, phenazine, phenoxathiin, phenoxazine, phenothiazine, thianthrene, thioxanthene, xanthene, and the like.
  • heterocycles of this invention can be substituted with 1, 2, or 3 substituents independently selected from alkoxy, alkoxycarbonyl, alkyl, alkylcarbonyl, alkylcarbonyloxy, alkylthio, amido, amino, carboxy, cyano, formyl, halogen, haloalkyl, hydroxy, nitro, and oxo.
  • heterocyclealkyl refers to a heterocycle, as defined herein, appended to the parent molecular moiety through an alkyl group, as defined herein.
  • Representative examples of heterocyclealkyl include, but are not limited to, pyrid-3- ylmethyl, 2-pyrimidin-2-ylpropyl, and the like.
  • hydroxy refers to an -OH group.
  • hydroxy protecting group refers to a substituent which protects hydroxyl groups against undesirable reactions during synthetic or semisynthetic procedures.
  • hydroxy-protecting groups include, but are not limited to, substituted methyl ethers, for example, methoxymethyl, benzyloxyniethyl, 2- methoxyethoxymethyl, 2-(trimethylsiyll)-ethoxymethyl, benzyl, and triphenylmethyl; tetrahydropyranyl ethers; substituted ethyl ethers, for example, 2,2,2-trichloroethyl and t- butyl; silyl ethers, for example, trimethyisilyl, t-butyldimethylsilyl and t- butyldiphenylsilyl; cyclic acetals and ketals, for example, methylene acetal, acetonide and benzylidene acetal; cyclic ortho
  • mamal as used herein, has its ordinary meaning and includes human beings.
  • methyl refers to a -CH-, group.
  • -NR 82 R 83 refers to two groups, R 82 and R 83 , independently selected from the group consisting of hydrogen, alkylcarbonyl, and formyl, appended to the molecular moiety through a nitrogen atom. At least one of R 82 and R 83 must be other than hydrogen.
  • Representative examples of -NR 82 R 83 include, but are not limited to, methylcarbonylamino, formylamino, ethylcarbonyl(formyl)amino, and the like.
  • NR 82 R 83 alkylene refers to a -NR 82 R 83 group, as defined herein, appended to the parent molecular moiety through an alkylene group, as defined herein.
  • Representative examples of NR 82 R 83 alkylene include, but are not limited to, 2-(methylcarbonylamino)ethyl, 2-(formylamino)ethyl, 3-(ethylcarbonylamino)propyl 2-[ethylcarbonyl(formyl)amino]ethyl, and the like.
  • nitro refers to a -NO 2 group.
  • salts refers to those carboxylate salts, amino acid addition salts, esters, amides and prodrugs of the compounds of the present invention which are, within the scope of sound medical judgement, suitable for use in contact with with the tissues of humans and lower animals with undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the invention.
  • salts refers to the relatively non-toxic, inorganic and organic acid addition salts of compounds of the present invention.
  • salts can be prepared in situ during the final isolation and purification of the compounds or by separately reacting the purified compound in its free base form with a suitable organic or inorganic acid and isolating the salt thus formed.
  • Representative salts include the hydrobromide, hydrochloride. sulfate, bisulfate, phosphate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate. fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactiobionate and laurylsulphonate salts and the like.
  • alkali and alkaline earth metals such as sodium, lithium, potassium, calcium, magnesium and the like
  • nontoxic ammonium, quaternary ammonium and amine cations including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine and the like.
  • ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine and the like See. for example S. M. Berge, et al., "Pharmaceutical Salts," J. Pharm. ScL, 66: 1-19 (1977) which is inco ⁇ orated herein by reference.)
  • esters of the compounds of this invention include C, to C 6 alkyl esters wherein the alkyl group is a straight or branched chain. Acceptable esters also include C 5 to C 7 cycloalkyl esters as well as arylalkyl esters such as, but not limited to benzyl. Preferred alkyl esters are C, to C 4 . Esters of the compounds of the present invention may be prepared according to conventional methods.
  • Examples of pharmaceutically acceptable, non-toxic amides of the compounds of this invention include amides derived from ammonia, primary C, to C 6 alkyl amines and secondary C, to C 6 dialkyl amines wherein the alkyl groups are straight or branched chain. In the case of secondary amines the amine may also be in the form of a 5 or 6 membered heterocycle containing one nitrogen atom. Amides derived from ammonia, C, to C 3 alkyl primary amides and C, to C 2 dialkyl secondary amides are preferred. Amides of the compounds of the invention may be prepared according to conventional methods.
  • prodrug refers to compounds that are rapidly transformed in vivo to yield the parent compound of the above formula, for example by hydrolysis in blood.
  • a thorough discussion is provided in T. Higuchi and V. Stella, "Pro-drugs as Novel Delivery Systems", Vol 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press,
  • prodrugs of derivatives of compounds of the present invention may be prepared by any suitable method.
  • the condensation of the amino group with amino acids and peptides may be effected in accordance with conventional condensation methods such as the azide method, the mixed acid anhydride method, the DCC (dicyclohexylcarbodiimede) method, the active ester method (p-nitrophenyl ester method, N-hydroxysuccinic acid imide ester method, cyanomethyl ester method and the like), the Woodward reagent K method, the DCC-HOBT (1-hydroxy-benzotriazole) method and the like.
  • Classical methods for amino acid condensation reactions are described in "Peptide
  • the irnmunosuppressant activity of the compounds of the present invention was determined using the human mixed lymphocyte reaction (MLR) assay described by Kino, T. et al., in Transplantation Proceedings, XIX(5):36-39, Suppl. 6 (1987), hereby inco ⁇ orated by reference.
  • MLR human mixed lymphocyte reaction
  • Lewis rats obtained from Charles River, Wilmington, MA
  • weighing 225-250 grams were acclimated for one week in a AAALAC approved facility. All animals were fasted overnight prior to dosing and throughout the study period but were permitted free access to water. Groups of animals were given FK-506 or Example 22 at a dose of 5 mg/kg.
  • Drugs were given in a 2 ml/kg volume of 10% ethanol, 40% propylene glycol and 2%> cremophore in 5% dextrose solution for both oral (p.o.) and intraperitoneal (i.p.) administration, and in a 1 ml/kg volume of the same vehicle without cremophor for intravenous (i.v.) dosing. Blood samples were collected from the tail vein in heparinized tubes at selected time points 0.25, 2.0, and 4.0 hours after dosing.
  • Drugs were separated from the hemolyzed whole blood contaminants utilizing liquid-liquid extraction with ethyl acetate :hexane (1 :1 by volume). Samples were centrifuged at 1200 x g for 10 minutes (4 °C) and a constant volume of the organic layer was transferred to a conical centrifuge tube and evaporated to dryness with a gentle stream of dry air over low heat ( ⁇ 35 °C). The samples were reconstituted with 40% (v/v) acetonitrile in water with vortexing.
  • the compounds of interest were separated from the co-extracted components by a 5 cm X 4.6 mm, 3 ⁇ m Spherisorb ODS-2 column (Regis, Morton Grove, IL) with an acetonitrile :methanol: 0.1% trifluoroacetic acid / 0.01 M tetramethylammonium perchlorate mixture (45:5:50 by volume for FK506, 40:5:55 for analogue Example 22) at a flow rate of 1.0 ml/min with UV detection at 205 nm.
  • the temperature of the HPLC column was maintained at 70 °C.
  • Example 22 was eliminated from the blood at 15 minutes. This result suggests that phosphates of the present invention may have reduced adverse systemic effects and therefore may be an advantage when a topical application is considered.
  • the compounds of the invention possess immunomodulatory activity in animals.
  • the compounds of the present invention may be useful for the treatment and prevention of immune-mediated diseases such as the resistance by transplantation of organs or tissue such as heart, kidney, liver, medulla ossium, skin, cornea, lung, pancreas, intestinum ***, limb, muscle, nervus, duodenum, small-bowel, pancreatic-islet-cell, etc.; graft- versus-host diseases brought about by medulla ossium transplantation; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, type I diabetes uveitis, allergic encephalomyelitis, glomerulonephritis, and the like; and further infectious diseases caused by pathogenic microorganisms.
  • immune-mediated diseases such as the resistance by transplantation of organs or tissue such as heart,
  • Further uses may include the treatment and prophylaxis of inflammatory and hype ⁇ roliferative skin diseases and cutaneous manifestations of immunologically-mediated illnesses, such as psoriasis, atopical dermatitis, contact dermatitis and further eczematous dermatitises, seborrhoeis dermatitis, Lichen planus,
  • Pemphigus bullous pemphigoid, Epidermolysis bullosa, urticaria, angioedemas, vasculitides, erythemas, cutaneous eosinophilias, Lupus erythematosus, acne and Alopecia areata; various eye diseases (autoimmune and otherwise) such as keratoconjunctivitis, vernal conjunctivitis, uveitis associated with Behcet's disease, keratitis, he ⁇ etic keratitis, conical cornea, dystrophia epithelialis corneae, corneal leukoma, ocular pemphigus.
  • eye diseases autoimmune and otherwise
  • reversible obstructive airway disease which includes condition such as asthma (for example, bronchial asthma, allergic asthma, intrinsic asthma, extrinsic asthma and dust asthma), particularly chronic or inveterate asthma (for example, late asthma and airway hyper-responsiveness), bronchitis and the like; inflammation of mucosa and blood vessels such as gastric ulcers, vascular damage caused by ischemic diseases and thrombosis, ischemic bowel diseases, inflammatory bowel diseases, necrotizing enterocolitis, intestinal lesions associated with thermal burns and leukotriene B 4 -mediated diseases; intestinal inflammations/allergies such as Coeliac diseases, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease and ulcerative colitis; food-related allergic diseases which have symptomatic manifestation remote
  • Goodpasture's syndrome, hemolytic-uremic syndrome and diabetic nephropathy nervous diseases such as multiple myositis, Guillain-Barre syndrome, Meniere's disease, polyneuritis, multiple neuritis, mononeuritis and radiculopathy; endocrine diseases such as hyperthyroidism and Basedow's disease; hematic diseases such as pure red cell aplasia, aplastic anemia, hypoplastic anemia, idiopathic thrombocytopenic pu ⁇ ura, autoimmune hemolytic anemia, agranulocytosis, pernicious anemia, megaloblastic anemia and anerythroplasia; bone diseases such as osteoporosis; respiratory diseases such as sarcoidosis, fibroid lung and idiopathic interstitial pneumonia; skin disease such as dermatomyositis, leukoderma vulgaris.
  • ichthyosis vulgaris photoallergic sensitivity and cutaneous T cell lymphoma
  • circulatory diseases such as arteriosclerosis, atherosclerosis, aortitis syndrome, polyarteritis nodosa and myocardosis; collagen diseases such as scleroderma, Wegener's granuloma and Sjogren's syndrome; adiposis; eosinophilic fasciitis; periodontal disease such as lesions of gingiva, periodontium, alveolar bone and substantia ossea dentis; nephrotic syndrome such as glomerulonephritis; male pattern aleopecia or alopecia senilis by preventing epilation or providing hair germination and/or promoting hair generation and hair growth; muscular dystrophy; Pyoderma and Sezary's syndrome; Addison's disease; active oxygen-mediated diseases, as for example organ injury such as ischemia-reperfusion injury of organs (such as heart, liver, kidney
  • the compounds of the invention are useful for the treatment and prevention of hepatic disease such as immunogenic diseases (for example, chronic autoimmune liver diseases such as the group consisting of autoimmune hepatitis, primary biliary cirrhosis and sclerosing cholangitis), partial liver resection, acute liver necrosis (e.g. necrosis caused by toxin, viral hepatitis, shock or anoxia), B-virus hepatitis, non-
  • immunogenic diseases for example, chronic autoimmune liver diseases such as the group consisting of autoimmune hepatitis, primary biliary cirrhosis and sclerosing cholangitis
  • partial liver resection for example, acute liver necrosis (e.g. necrosis caused by toxin, viral hepatitis, shock or anoxia), B-virus hepatitis, non-
  • A/non-B hepatitis, cirrhosis (such as alcoholic cirrhosis) and hepatic failure such as fulminant hepatic failure, late-onset hepatic failure and "acute-on-chronic" liver failure (acute liver failure on chronic liver diseases), and moreover are useful for various diseases because of their useful activity such as augmention of chemotherapeutic effect, preventing or treating activity of cytomegalovirus infection, particularly HCMV infection, anti- inflammatory activity, and so on.
  • the compounds of the present invention may thus be used in the treatment of immunodepression or a disorder involving immunodepression.
  • disorders involving immunodepression include AIDS, cancer, senile dementia, trauma (including wound healing, surgery and shock) chronic bacterial infection, and certain central nervous system disorders.
  • the immunodepression to be treated may be caused by an overdose of an immunosuppressive macrocyclic compound, for example derivatives of 12-(2- cyclohexyl-l-methylvinyl)-13, 19,21,27-tetramethyl-l l,28-dioxa-4-azatricyclo[22.3.1.0 ' 9 ] octacos- 18-ene such as FK-506. or rapamycin. Overdosing of such medicants by patients is quite common upon their realizing that they have forgotten to take their medication at the prescribed time and can lead to serious side effects.
  • a further situation in which the compounds of the present invention may be used to treat immunosuppression is in vaccination. It is sometimes found that the antigen introduced into the body for the acquisition of immunity from disease acts as an immunosuppressive agent, and so antibodies are not produced by the body and immunity is not acquired. By introducing a compound of the invention into the body (as in a vaccine), the undesired immunosuppression may be overcome and immunity acquired.
  • Aqueous liquid compositions of the present invention may be particularly useful for the treatment and prevention of various diseases of the eye such as autoimmune diseases (including, for example, conical cornea, keratitis, dysophia epithelialis corneae, leukoma, Mooren's ulcer, sclevitis and Graves' ophthalmopathy) and rejection of corneal transplantation.
  • autoimmune diseases including, for example, conical cornea, keratitis, dysophia epithelialis corneae, leukoma, Mooren's ulcer, sclevitis and Graves' ophthalmopathy
  • a therapeutically effective amount of one of the compounds of the present invention may be employed in pure form or, where such forms exist, in pharmaceutically acceptable salt, ester or prodrug form.
  • the compound may be administered as pharmaceutical compositions containing the compound of interest in combination with one or more pharmaceutically acceptable excipients.
  • a "therapeutically effective amount" of the compound of the invention is meant a sufficient amount of the compound to treat gastrointestinal disorders, at a reasonable benefit/risk ratio applicable to any medical treatment. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgement.
  • the specific therapeutically effective dose level for any particular patient will depend upon a variety of factors including the disorder being treated and the severity of the disorder; activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed; and like factors well known in the medical arts. For example, it is well within the skill of the art to start doses of the compound at levels lower than required for to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved.
  • the total daily dose of the compounds of this invention administered to a human or lower animal may range from about 0.001 to about 3 mg/kg/day.
  • more preferable doses may be in the range of from about 0.005 to about
  • the effective daily dose may be divided into multiple doses for pu ⁇ oses of administration; consequently, single dose compositions may contain such amounts or submultiples thereof to make up the daily dose.
  • compositions of the present invention comprise a compound of the invention and a pharmaceutically acceptable carrier or excipient, which may be administered orally, rectally, parenterally, intracistemally, intravaginally. intraperitoneally, topically (as by powders, ointments, drops or transdermal patch), bucally, or as an oral or nasal spray.
  • pharmaceutically acceptable carrier is meant a non-toxic solid, semi- solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type.
  • parenteral refers to modes of administration which include intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous and intraarticular injection and infusion.
  • compositions of this invention for parenteral injection comprise pharmaceutically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions as well as sterile powders for reconstitution into sterile injectable solutions or dispersions just prior to use.
  • suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol. and the like), carboxymethylcellulose and suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters such as ethyl oleate.
  • Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.
  • compositions may also contain adjuvants such as preservative, wetting agents, emulsifying agents, and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents such as sugars, sodium chloride, and the like, Prolonged abso ⁇ tion of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay abso ⁇ tion such as aluminum monostearate and gelatin. In some cases, in order to prolong the effect of the drug, it is desirable to slow the abso ⁇ tion of the drug from subcutaneous or intramuscular injection.
  • adjuvants such as preservative, wetting agents, emulsifying agents, and dispersing agents.
  • Injectable depot forms are made by forming microencapsule matrices of the drug in biodegradable polymers such as polylactide-polyglycolide. Depending upon the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides) Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues.
  • biodegradable polymers such as polylactide-polyglycolide.
  • Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues.
  • the injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by inco ⁇ orating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium just prior to use.
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.
  • the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol. and silicic acid, b) binders such as, for example, carboxymethylcellulose.
  • the dosage form may also comprise buffering agents.
  • compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.
  • the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes.
  • the active compounds can also be in micro-encapsulated form, if appropriate, with one or more of the above-mentioned excipients.
  • Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs.
  • the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethyl formamide, oils (in particular, cottonseed, groundnut, com, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.
  • inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers
  • the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
  • adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
  • Suspensions in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, and mixtures thereof.
  • Topical administration includes administration to the skin or ucosa, including surfaces of the lung and eye.
  • Compositions for topical administration, including those for inhalation, may be prepared as a dry powder which may be pressurized or non-pressurized.
  • the active ingredient in finely divided form may be used in admixture with a larger-sized pharmaceutically acceptable inert carrier comprising particles having a size, for example, of up to 100 micrometers in diameter.
  • suitable inert carriers include sugars such as lactose.
  • at least 95% by weight of the particles of the active ingredient have an effective particle size in the range of 0.01 to 10 micrometers.
  • the composition may be pressurized and contain a compressed gas, such as nitrogen or a liquified gas propellant.
  • the liquified propellant medium and indeed the total composition is preferably such that the active ingredient does not dissolve therein to any substantial extent.
  • the pressurized composition may also contain a surface active agent.
  • the surface active agent may be a liquid or solid non-ionic surface active agent or may be a solid anionic surface active agent. It is preferred to use the solid anionic surface active agent in the form of a sodium salt.
  • a further form of topical administration is to the eye, as for the treatment of immune-mediated conditions of the eye such as automimmue diseases, allergic or inflammatory conditions, and corneal transplants.
  • the compound of the invention is delivered in a pharmaceutically acceptable ophthalmic vehicle, such that the compound is maintained in contact with the ocular surface for a sufficient time period to allow the compound to penetrate the comeal and internal regions of the eye.
  • a pharmaceutically acceptable ophthalmic vehicle may, for example, be an ointment, vegetable oil or an encapsulating material.
  • compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non- irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at room temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
  • suitable non- irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at room temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
  • Compounds of the present invention can also be administered in the form of liposomes.
  • liposomes are generally derived from phospholipids or other lipid substances. Liposomes are formed by mono- or multi-lamellar hydrated liquid crystals that are dispersed in an aqueous medium.
  • any non-toxic, physiologically acceptable and metabolizable lipid capable of forming liposomes can be used.
  • the present compositions in liposome form can contain, in addition to a compound of the present invention, stabilizers, preservatives, excipients, and the like.
  • the preferred lipids are the phospholipids and the phosphatidyl cholines (lecithins), both natural and synthetic. Methods to form liposomes are known in the art. See. for example, Prescott, Ed., Methods in Cell Biology, Volume XIV, Academic Press, New York, N.Y. (1976), p. 33 et seq.
  • HC1 for hydrochloric acid
  • THF for tetrahydrofuran
  • TFA for trifluoroacetic acid
  • MgSO 4 for magnesium sulfate
  • Na 2 SO 4 for sodium sulfate
  • NaCl for sodium chloride
  • NaHCO 3 for sodium bicarbonate
  • NH 4 C1 for ammonium chloride
  • HPLC high pressure liquid chr ⁇ matography
  • PPTS pyridinium p-toluensulfonate.
  • the compounds of the present invention may be prepared using one or more of the processes which follow.
  • the starting materials for use in these processes are preferably one of the macrolides isolated from culture media obtained in accordance with known methods by fermentation of microorganisms of the genus Streptomyces, which are disclosed in European Patent Application No. 0184162. Samples are available from the Fermentation Research Institute, Tsukuba, Ibaraki 305, Japan under the provisions of the Budapest Treaty, under deposit No. FERM BP-927. This strain has been redeposited on
  • FR-900520 European Patent Application 0184162
  • the macrolide FR-900520 may be prepared in accordance to the published methods of (i) H. Hatanaka, M. Iwami, T. Kino, T. Goto and M. Okuhara, FR-900520 and FR-900523,
  • the compounds of this invention can be prepared by a variety of synthetic routes. Representative procedures are shown in Schemes 1-4. For Schemes 1-4, a broken line represents an optional double bond, R 1 , R 2 , R 3 , and R 4 are as defined in formula I.
  • the hydroxy group attached to carbon 4' of formula (i) can be phosphorylated with a phosphoramidochloridoite (ii), wherein X is a secondary amine and R is selected from alkyl, cyanoalkyl, haloalkyl. and phenyl, to give phosphoramidoites of formula (iii).
  • Suitable phosphorylating reagents include, but are not limited to, N,N- diisopropylmethyl phosphonamidic chloride (Bruzik, K.S., Salamonczyk, G. and Stec, W.J. J. Org.
  • reaction is preferrably conducted in the presence of an organic base such as trialkylamines (e.g. triethylamine, etc.), pyridine compounds (e.g.
  • pyridine lutidine, picoline, 4-N,N-dimethylaminopyridine, etc.
  • preferred bases are triethylamine, diisopropylethylamine, and pyridine.
  • phosphoramidoates or phosphoramidothioatesof formula (iv), wherein Z is O or S respectively can be prepared by oxidation of phosphoramidoites of formula (iii).
  • the procedure described by Hecker, S.J., Minich, M.L. and Lackey, K., J. Org. Chem., (1990), 55, 4904-491 1) can be used to give phosphoramidoates and elemental sulphur can give the phosphoramidothioates.
  • phosphites of formula (v) can be prepared by treating phosphoramidoites of formula (iii) with an alcohol or phenol in the presence of tetrazole. Conversion of phosphoramidoites to phosphites utilizing lH-tetrazole is described in (Beaucage, S.L. and Iyer, R.P. Tetrahedron, (1993), 49, 6123-6194; Reese, C.B. Tetrahedron, (1978), 34, 3143-3179; Liu, J. and Verdine, G.L. Tetrahedron Lett., (1992),
  • activators that can be used for the conversion of phosphoramidoites to phosphites include, but are not limited to, additional members of the tetrazole class of activators: 5-(p- nitrophenyl) tetrazole (Froehler, B. C. and Mattcucci, M. D., Tetrahedron Letters (1983), 24, 3171 -3174); 5-(p-nitrophenyl) tetrazole and DMAP (Pon, R.T., Tetrahedron Letters
  • phosphites of formula (v) can be oxidized to phosphates or thiophosphates of formula (vi), wherein Z is O or S respectively. Oxidation of phosphites to phosphates can be accomplished using per acids such as tert-butylhydroperoxide
  • oxidizing agents include iodine in aquous tetrahydrofuran at low temperature (Lestinger, R.L. and Lunsford, W.B. J. Am. Chem. Soc, (1976), 98, 3655).
  • Formation of a thiophosphate from a phosphite may be carried out using elemental sulfur (Bruzik, K.S., Salamonczyk, G. and Stec, W.J., J. Org. Chem., (1986). 51. 2368- 2370; Stec, W.J., Grajkowski, A., Koziolkiewicz. M. and Uznanski, B., Nucleic Acid Res., (1991), 19, 5883-5888; Martin, S.F. and Wagman, A.S., J. Org. Chem., (1993), 58, 5897-5899).
  • the hydroxy group at the C-l 4 position of formula (i) may or may not be necessary to protect.
  • suitable protecting groups include, but are not limted to, dimefhylthexylsilyl, trisubstituted silyl such as tri(lower)alkylsilyl (e.g. trimethylsilyl, triethylsilyl, tributylsilyl, tri-i-propylsilyl, tert-butyl-dimethylsilyl, tri-tert-butylsilyl, triphenylmethyl-dimethylsilyl, etc.); lower alkyldiarylsilyl (e.g.
  • triarysilyl e.g. triphenylsilyl, tri-p-xylylsilyl. etc.
  • triarylalkylsilyl e.g. tribenzylsilyl, etc.
  • the preferred one may be tri(C,-C
  • Suitable o-silylations may be carried out using a wide variety of organosilicon reagents including, but not limitted to, tert-butyldimethylsilyl chloride, N- (tert-butyldimethylsilyl)-N-methyltrifluoroacetamide ( Mawhinney, T., and Madison, M.
  • Carbonate hydroxy-protecting groups may also be used and introduced using a wide variety of a corresponding helocarbonate compound, such as methyl chloroformate, ethyl chloroformate, 2,2,2-trichloroethyl chloroformate, isobutyl chloroformate, vinyl chloroformate, allyl chloroformate, benzyl or substituted benzyl chloroformate such as p-methoxybenzyl, 3,4-dimethylbenzyl, p- nitrobenzyl etc., 2-(trimethylsilyl)ethyl chloroformate, 2-(benzenesulfonyl)ethyl chloroformate, 2-(trimethylsilyl)ethoxymethyl chloride and the like.
  • a corresponding helocarbonate compound such as methyl chloroformate, ethyl chloroformate, 2,2,2-trichloroethyl chloroformate, isobutyl chloroformate, vinyl
  • tertiary base such as pyridine, triethylamine, imidazole, diisopropylethylamine, and the like.
  • suitable reagents for deprotection include, but are not limited to aqueous hydrogen fluoride in acetonitrile (Newton, R. F., Reynolds, D. P., Finch, M. A. W., Kelly, D. R. and Roberts, S. M. Tetrahedron Lett., (1979), 3891), tetraalkyl ammonium fluoride in tetrahydrofuran (Corey, E. J. and Snider,
  • N,N-dimethylformamide 500 mL
  • tert-butyldimethylchlorosilane 47.64 g, 0.32 mol
  • Dimethylformamide and excess tert-butyldimethylchlorosilane were removed by evaporation (35 °C water bath ) under high vaccum.
  • the solid residue was dissolved in 350 mL of ethyl acetate, and the ethyl acetate layer was washed in succession with saturated NH 4 C1 solution (200 mL X3), 10% NaHSO solution(200 mL X3), brine, saturated NaHCO 3 solution (200 mL X3), brine (200 mL X3), and dried (MgSO 4 ).
  • the reaction mixture was cooled to 0 °C in an ice bath and treated with solid NaHCO, with stirring for 1 hour.
  • the solids were removed by filtration and the acetonitrile was removed in vacuo.
  • the residue was taken up with ethyl acetate (500 mL) and washed in succession with 10% NaHCO 3 solution (300 mL X3), brine (250 mL), 10% NaHSO 4 solution (300 mL X3). brine ( 350 mL X3), and dried (NaSO 4 ).
  • minor 67.3 (s, pyridyl methyl, minor), 67.1 (s, pyridyl), 57.8 (t, minor), 57.6 (t), 57.1 (t), 57.0 (t, major and minor), 56.2 (t), 55.9 (t, minor), 55.5 (t), 55.3 (t, minor), 54.6, (p, P-OCH3), 54.3, (p, P-OCH3, minor), 53.1 (t), 49.4 (s), 48.0 (s, minor), 48.0 (s), 46.6 (s.
  • CH 2 , CH 2 . and CH 2 1.62 (s, CH 3 ), 1.79 (s, CH 3 , minor). 1.80 (s, CH 3 ), 1.86 (s, CH 3 , minor), 1.98 (m, CH), 2.10 (m, CH 2 , CH 2 , and CH), 2.22 (m, CH 2 ), 2.31, 2.35 (m, CH 2 and CH).
  • Example 8 (lR,2R.4R)-4- E)-2-r(lR.9S.12S.13R.14S.17R.21S.23S.24R.25S.27R)-17-ethyl-1.14- dihydroxy-23 ,25-dimethoxy- 13.19.21.27-tetramethyl-2.3.10.16-tetraoxo- 11 ,28-dioxa-4- azatricyclo[22.3.1.0 49 ]octacos-l 8-en-l 2-yl]- 1 -propenyl ⁇ -2-methoxycyclohexyl 2- furylmethyl methyl phosphate
  • Example 9 benzyl (lR,2R.4R)-4-((E)-2-r(lR.9S.12S.13R.14S.17R.21 S.23S.24R.25S.27R)-17-ethyl- 1.14-dihvdroxy-23.25-dimethoxy-13.19.21.27-tetramethyl-2.3.10.16-tetraoxo-11.28- dioxa-4-azatricyclo[22.3.1.0 49 ]octacos-l 8-en-l 2-yl1-l-propenyl ⁇ -2-methoxycyclohexyl methyl phosphate
  • phenylmethanol (0.310 mL, 3 mmol)
  • Example 10 (lR.2R,4R)-4- ⁇ (EV2-r(lR.9S.12S.13R.14S.17R.21S.23S.24R,25S.27R)-17-ethyl-1.14- dihydroxy-23.25-dimethoxy- 13.19.21.27-tetramethyl-2.3.10.16-tetraoxo- 11.28-dioxa-4- azatricyclo[22.3.1.0 4 9 ]octacos-l 8-en- 12-yll-l -propenyl j-2-methoxycvclohexyl 3- furylmethyl methyl phosphate
  • 3-furylmethanol (258 mL, 3 mmol) and the procedure described in Example 5 were used to provide the title compound.
  • Example 1 1 (lR.2R.4R)-4- ⁇ (E)-2-[(lR.9S.12S.13R.14S.17R.21 S.23S.24R.25S.27RV17-ethyl-1.14- dihvdroxy-23.25-dimethoxy-13.19.21.27-tetramethyl-2.3.10.16-tetraoxo-11.28-dioxa-4- azatricvclo[22.3.1.0 9 ]octacos-l 8-en-l 2-yl]- 1 -propenyl) -2-methoxycyclohexyl methyl 3- thienylmethyl phosphate
  • the product from Example 3 (1.067 g, 1 mmol), 3-fhienylmethanol (142 ⁇ L, 1.5 mmol) and the procedure described in Example 5 were used to provide (61.6 mg) the title compound.
  • Example 12 (lR.2R.4R)-4- ⁇ (E -2-[(lR.9S.12S.13R.14S.17R,21 S.23S.24R.25S,27R)-17-ethyl-1.14- dihydroxy-23.25-dimethoxy- 13.19.21.27-tetramethyl-2.3, 10.16-tetraoxo- 1 1 ,28-dioxa-4- azatricyclo[22.3.1.0 49 ]octacos-l 8-en-l 2-yl]- 1 -propenyl) -2-methoxycyclohexyl 4- fluorobenzyl methyl phosphate
  • Example 13 ( 1 ,3-dioxo- 1.3 -dihvdro-2H-isoindol-2-yl)methv 1 ( 1 R.2R.4R)-4- ⁇ (E)-2- r(lR.9S.12S.13R,14S,17R,21 S,23S,24R.25S.27R)-17-ethyl-1.14-dihvdroxy-23.25- dimethoxy- 13.19.21.27-tetramethyl-2.3.10.16-tetraoxo- 1 1.28-dioxa-4- azatricyclo[22.3.1.0 4 ' 9 ]octacos-l 8-en-l 2-yl]- 1 -propenyl )-2-methoxycvclchexyl methyl phosphate
  • Example 14 (lR.2R.4R)-4- ⁇ (EV2-[(lR,9S.12S.13R.14S.17R.21 S.23S.24R.25S.27R)-17-ethyl-1.14- dihvdroxy-23,25-dimethoxy- 13.19.21.27-tetramethyl-2.3.10.16-tetraoxo- 1 1.28-dioxa-4- azatricyclo[22.3.1.0 4,9 ]octacos- 18-en- 12-yl]- 1 -propenyl ) -2-methoxycyclohexyl methyl 4- nitrobenzyl phosphate The product from Example 3.
  • Example 15 (lR.2R.4R)-4- ⁇ (E)-2-r(lR.9S.12S.13R.14S.17R.21 S.23S.24R.25S.27R -17-ethyl-1.14- dihvdroxy-23.25-dimethoxy- 13.19,21.27-tetramethyl-2.3.10.16-tetraoxo- 11.28-dioxa-4- azatricyclo[22.3.1.0 49 ]octacos- 18-en- 12-yl]- 1 -propenyl ) -2-methoxycyclohexyl methyl diisopropylphosphoramidoate
  • the product from Example 3 was oxidized according to the method described in (Hecker, S.
  • Example 16 2-cvanoethyl (lR.2R,4R)-4- ⁇ (E)-2-[(lR,9S,12S,13R.14S.17R.21 S.23S.24R,25S.27R)-17- ethyl-1.14-dihydroxy-23.25-dimethoxy-13.19.21.27-tetramethyl-2.3.10.16-tetraoxo-11.28- dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos- 18-en- 12-yl]- 1 -propenyl ) -2-methoxycyclohexyl diisopropylphosphoramidoate
  • the product from Example 4 was oxidized according to the method described in (Hecker, S.
  • Example 3 The product from Example 3 (1.067 g, 1 mmol), (3-iodophenyl)methanol (330 mg, 1.5 mmol), and the procedure described in Example 5 were used to provide (280.5 mg) the title compound.
  • MS (FAB) m/z: M+K + l 126; ⁇ C NMR (500 MHz in Pyridine-d 5 ) ⁇ 211.6 (q), 210.7 (q, minor), 199.6 (q, minor), 198.8 (q), 170.1 (q, minor), 170.0 (q), 167.3 (q, minor), 167.0 (q). 139.3 (q. minor), 138.4 (q),
  • CH 3 1.86 (s, CH 3 , minor), 1.98 (m, CH), 2.09 (s, acetoamidoCH 3 ), 2.10 (m, CH 2 , CH 2 , and CH), 2.22 (m, CH 2 ), 2.31, 2.35 (m, CH 2 and CH), 2.92 (m, CH 2 , minor), 2.57 (m. CH), 2.67 (m, CH), 2.77 (m, CH, minor), 2.91 (m, CH, minor), 3.02 (m, CH, minor), 3.08 (m, CH), 3.23 (m, CH 2 ), 3.3 (m, CH), 3.36 (s.
  • OCH 3 , minor 3.38 (s, OCH 3 ), 3.42 (s, OCH 3 , major and minor), 3.44 (s, OCH 3 ), 3.45 (s, OCH 3 , minor), 3.55 (m, CH major and minor). 3.62 (m, CH?, minor).
  • Example 21 (lR,2R,4R)-4- ⁇ (E)-2-r(lR,9S.12S.13R,14S.17R.21 S,23S,24R.25S.27R -17-ethyl-1.14- dihydroxy-23.25-dimethoxy- 13.19,21.27-tetramethyl-2.3.10,16-tetraoxo- 11 ,28-dioxa-4- azatricyclo [22.3.1.0 4 9 ]octacos- 18-en- 12-yl] - 1 -propenyl ) -2-methoxycvclohexyl 4- fluorophenyl methyl phosphate
  • the product from Example 3, 4-fluorophenol, and the procedure described in Example 5 were used to provide the title compound.
  • Example 22 (lR.2R.4R)-4- ⁇ (E)-2-KlR.9S,12S.13R.14S.17R.21 S.23S.24R.25S.27R)-17-ethyl-1.14- dihvdroxy-23.25-dimethoxy- 13.19.21 ,27-tetramethyl-2.3.10.16-tetraoxo- 1 1 ,28-dioxa-4- azatricyclo[22.3.1.0 49 1octacos- 18-en- 12-yl]- 1 -propenyl ) -2-methoxycyclohexyl 4- formylphenyl methyl phosphate
  • a solution of the product from Example 3 (1.600 g, 1.5 mmol) in tetrahydrofuran : acetonitrile (1 :1) (30 mL) under a nitrogen atmosphere was treated with 4- hydroxybenzaldehyde(274.8 mg, 2.25 mmol)
  • tert-butylhydroperoxide (0.122 mL, 3 mmol, in 2,2,4- trimethylpentane) was directly added to the reaction mixture and allowed to stir at ambient temperature overnight. The solvents were removed and the residue was dissolved in ethyl acetate (50 mL), quickly washed in succession with 10%) sodium bicarbonate, brine, and dried (MgSO 4 ). The crude product (1.68 g) was dissolved in acetonitrile (45 mL) and treated with 48% hydrofluoric acid (1.5 mL). After stirring for 30 minutes, the mixture was cooled and ethyl acetate and cold 10% sodium hydrogen sulfate were carefully added to the reaction mixture.
  • reaction mixture was quenched with 10% NaHSO 4 (20 ml) and partitioned with ethyl acetate (40 ml).
  • the organic layer was successively washed with 10% NaHSO 4 (20 ml, X2), saturated NaHCO 3 (20 ml, X2), brine (20 ml, X3), and dried (Na 2 SO 4 ).
  • Methylsulfi de-chlorine complex was prepared by adding oxalyl chloride
  • Phosphate analogs wherein R 6 is -OP(O)AB and R' is hydrogen can be prepared using the product from Example 27 and the procedures described in Examples 1-25.

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EP00918274A 1999-03-31 2000-03-23 Phosphat-enthaltende makrozyclische immunomodulatoren Withdrawn EP1165575A1 (de)

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CA2103453A1 (en) * 1991-09-05 1993-03-06 Jay R. Luly Macrocyclic immunomodulators
US5457111A (en) * 1991-09-05 1995-10-10 Abbott Laboratories Macrocyclic immunomodulators
US5457194A (en) * 1993-03-17 1995-10-10 Abbott Laboratories Substituted aliphatic amine-containing macrocyclic immunomodulators

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