CN110835345A

CN110835345A - Degradation agent of cell cycle dependent kinase, preparation method thereof, pharmaceutical composition and application thereof

Info

Publication number: CN110835345A
Application number: CN201810943251.1A
Authority: CN
Inventors: 陈小华; 李佳; 冯序乐; 周宇波; 田洪涛; 张凯祥; 王培培
Original assignee: Shanghai Institute of Materia Medica of CAS
Current assignee: Shanghai Institute of Materia Medica of CAS
Priority date: 2018-08-17
Filing date: 2018-08-17
Publication date: 2020-02-25
Also published as: CN111065639B; CN111065639A; WO2020035049A1

Abstract

The invention relates to a target cell Cycle Dependent Kinase (CDKs) degradation agent shown as a general formula (I)Compounds and/or pharmaceutically acceptable salts thereof, pharmaceutical compositions thereof and the use of these derivatives as pharmaceutically active agents in medicaments for the prevention and/or treatment of diseases associated with abnormal activity of CDKs. The compound has better effect of inhibiting cell proliferation on solid tumor and blood tumor, and shows potential for treating related cancers and autoimmune diseases.

Description

Degradation agent of cell cycle dependent kinase, preparation method thereof, pharmaceutical composition and application thereof

Technical Field

The invention belongs to the field of pharmacology, and particularly relates to a compound shown as a general formula (I), a preparation method thereof, a pharmaceutical composition and application thereof in preparing a degradation agent of targeted cell Cycle Dependent Kinases (CDKs) in preventing and/or treating diseases or symptoms related to abnormal activity of the cell cycle dependent kinases and treating diseases or symptoms related to selective transcription of the CDKs. The invention also provides pharmaceutically acceptable compositions comprising the compounds of the invention and methods of using the compositions in the treatment of diseases or disorders associated with the selective transcription of CDKs.

Background

Important targets for cyclin-dependent kinase (CDK) tumor therapy:

the cell Cycle Dependent Kinases (CDKs) are serine/threonine kinases, and are mostly heterodimer complexes consisting of cell cycle catalytic kinase subunits and regulatory subunits, which are activated by binding with cyclin to regulate cell cycle and transcription processes, and the disorder of CDK activity directly or indirectly causes uncontrolled cell proliferation, increased DNA mutation, chromosome deletion, chromosome change and the like, resulting in the occurrence and development of tumors. CDK family proteins are involved in cellular mitosis and transcription processes and play an important role in cell proliferation, and since tumor cell proliferation is abnormally active, abnormal activation of CDKs is often observed in tumor cells, thereby promoting growth of tumor cells. The CDK family has been considered as important targets in tumor therapy, but since the amino acid sequences at the position of ATP binding pockets between different CDK proteins are highly conserved, it is difficult to find small molecule inhibitors with high selectivity between CDK families. Currently marketed drugs targeting the CDK family include bevacizumab (Palbociclib) marketed in 2015, norfloxacin (Ribociclib) marketed in 2017 and celecoxib, all of which are inhibitors targeting CDK4/6 and are used for treating breast cancer, and most of the drugs targeting other targets of the CDK family are in clinical or preclinical research stages.

Of the CDK family, CDK7 and CDK9 have recently gained increasing attention from academia and various large pharmaceutical companies. CDK7 has dual functions of affecting the cell cycle and regulating the transcriptional process. CDK7 is the catalytic subunit of the CDK-activating kinase Complex (CAK), which consists of CDK7, cyclin H and MAT1, which can activate cell cycle-related CDK proteins, including CDK1, CDK2, CDK4, CDK6, by threonine phosphorylation, thereby affecting cell cycle progression. In transcriptional regulation, CDK7, which is part of the basic transcription factor TFIIH complex, can activate RNA polymerase ii by serine phosphorylating the C-terminal domain of RNA polymerase ii, thereby regulating RNA polymerase ii-mediated transcription initiation and extension.

CDK9/CyclinT1 (or T2) is part of the subunit of the positive transcription elongation factor b (P-TEFb) complex, which activates RNA polymerase II to initiate regulation of gene transcription elongation by phosphorylating the carboxy-terminal domain (CTD) of RNA polymerase. CDK9 is located upstream of cell cycle regulatory proteins such as anti-apoptotic proteins Bcl-2, Mcl-1, XIAP, etc. and critical molecules such as mitosis-regulated kinases, which are closely related to tumors, and can be directly or indirectly regulated by regulation of CDK9, such as: in Chronic Lymphocytic Leukemia (CLL), a therapeutic effect can be achieved by down-regulating key proteins of the Bcl-2 family by inhibiting the activity of CDK 9. CDK9 had no effect on cell cycle regulation, and this specific regulation of the transcriptional process by CDK9 made it one of the most potent anti-tumor targets in the CDK family. In addition, the documents report that CDK7/cyclin H and CDK9/cyclin T complexes also show that the complexes can inhibit the replication of RNA of HIV and HSV viruses, so that the complexes also have a certain application prospect in the field of antivirus.

Targeted protein degradation technology:

the targeted protein degradation technology is a new technology developed by a medicament which is widely concerned in recent years, and the technology utilizes a ubiquitin-proteasome mediated protein degradation pathway in organisms to artificially control the degradation of target proteins. By designing a bifunctional molecule, one end of the bifunctional molecule can recognize a target protein, the other end can recognize E3ubiquitin ligase, and the two ends are connected through a connecting chain (Linker) to form a target induced protein degradation complex (PROTACs). After the bifunctional molecule enters a cell, two ends can respectively recognize a target protein and E3ubiquitin ligase, the two proteins are drawn close to each other, ubiquitin on E2 ubiquitin ligase is transferred to the target protein, the target protein is ubiquitinated, and then the target protein is degraded through an ubiquitin-proteasome pathway. Through the rapid development of recent years, the targeted protein degradation technology has been changed from a chemical biological concept to a new technology for drug development, the defects of the traditional small molecule inhibitor can be overcome by utilizing the PROTAC molecule to target and degrade the target points related to diseases, and the small molecule drug is hopefully redefined. The documents CN103265635A, CN107257800A and the like describe the effect of the technology on degrading target proteins.

Protein degrading agents targeting CDK7 and CDK 9:

CDK7 and CDK9 are important targets for tumor therapy, and several CDK7 and CDK9 inhibitors have been currently in clinical study, most of which are non-selective inhibitors (j.med.chem.2016,59,8667-8684.Bioorganic & Medicinal chemistry letters2015, 25, 3420-. Representative small molecule inhibitors include Flavopiridol, Roscovitine, SNS-032, R547, TG-02, AT-7519, and the like; however, small molecule inhibitors with high selectivity against CDK7 or CDK9, or against dual targets of CDK7 and CDK9, have been less studied (US2007179161a1, US2015111873a1, WO2016193939a1, WO2016142855a 2). Although important methods for retarding cell cycle induction apoptosis and regulating transcription to become anti-tumor by acting on CDK7 and CDK9 through small molecule inhibitors, recent studies show that the small molecule inhibitors act on CDKs targets to generate drug resistance, and the treatment only through the small molecule inhibitors cannot become an effective anti-tumor method. SNS-032 is used as target protein ligand by Nathanael Grag and the like, and the general formula is developed

The degradant for targeted degradation of CDK9 of WO2017185023A1, wherein a representative one CDK9 degradant is THAL-SNS-032.

The CDK-related kinase is closely related to the occurrence and development of tumors, so that the degrading agent targeting CDK9 or CDK 7-related kinase targets can degrade target proteins, thereby regulating RNA polymerase II-mediated transcription initiation and extension and effectively inhibiting the proliferation of tumor cells; meanwhile, the drug resistance problem of the small molecule inhibitor of the target can be solved. It is therefore an object of the present invention to provide a class of compounds which may be used in the treatment and/or prevention or amelioration of such diseases and/or disorders by designing synthetic novel protein degrading agents targeting CDK9 or CDK 7.

Disclosure of Invention

The inventors of the present invention analyzed the following important information by analyzing the complex crystal structure of CDK9 and other CDKs (PDB ID: 1QMZ,3BLR, 1UA2, 3 BLH): CDK9 possesses greater conformational flexibility than other CDKs, such as small molecule inhibitors that, when bound to CDK9/cyclinT, can down-regulate the glycine (Gly) -rich loop positions of the CDK structure to accommodate ligand binding; at the same time, this feature of the CDK9 protein structure allows its binding domain to accommodate larger ligand structures. Through structural analysis, we also obtained important information in the complex structure of CDK9 with other small molecule inhibitors (PDB ID: 3LQ 5): the substituent on C-6 on the purine ring extends beyond the ATP-binding pocket of CDK9 and is partially exposed to solvent; and also maintains good binding to CDK9 if this position is a more sterically hindered substituent. Therefore, these characteristics will be the starting point for designing the CDK9 small molecule degradation agent. Based on the small molecule inhibitor of CDK9, the inventors mainly designed and synthesized a class of small molecule degraders specifically targeting CDK 9. By designing a bifunctional molecule, one end of the molecule can recognize a target protein, the other end can recognize E3ubiquitin ligase, and the middle is connected through a connecting chain (Linker). We synthesized a series of CDK9 degradants described herein and tested the activity of the compounds; the test result shows that the degradation agent has very high cell growth inhibition activity, and the degradation agent can degrade CDK9 target very efficiently through the related test method. After the molecule acts on an organism, the degradation of a target protein CDK9 can be artificially controlled through a ubiquitin-proteasome mediated protein degradation pathway in the organism, and further the effective disease treatment based on the CDK9 target point is realized.

An object of the present invention is to provide a compound represented by the general formula (I), a tautomer, an enantiomer, a diastereomer, a racemate, a metabolic precursor, a pharmaceutically acceptable salt, an ester, a prodrug, or a hydrate thereof.

The invention also aims to provide important intermediates of the compounds and a preparation method thereof.

The invention also aims to provide application of the compounds in preparing medicaments for preventing and treating diseases related to abnormal activity of CDKs, wherein the diseases related to the abnormal activity of CDKs comprise solid tumors such as breast cancer, colon cancer, prostatic cancer, small cell lung cancer, non-small cell lung cancer and the like, and blood tumors such as acute lymphocytic leukemia, B cell lymphoma, T cell lymphoma, myeloma, acute and chronic myelogenous leukemia, promyelocytic leukemia and the like.

The invention provides a compound shown as the following general formula (I), a tautomer, an enantiomer, a diastereomer, a racemate, a metabolic precursor, a pharmaceutically acceptable salt, an ester, a prodrug or a hydrate thereof:

wherein,

selected from the following groups:

the G is₁、G₂、G₃Each independently optionally from-C (R)_b1) or-N ═ or; wherein R is_b1Selected from hydrogen, halogen, or C_1-4A hydrocarbyl group;

z is-CH₂-or-C (═ O) -;

R_a1is hydrogen, fluoro or methyl;

R_a2is hydrogen or methyl;

R_c1is hydrogen or fluorine;

R_a3、R_a4、R_a5each independently hydrogen, halogen, cyano, nitro, hydroxy, amino, substituted or unsubstituted C_1-6Alkyl, substituted or unsubstituted C_1-6Alkoxy, substituted or unsubstituted amino, substituted or unsubstituted C_2-6Alkylcarbonyl, wherein said substitution means substitution with one or more of the following substituents: halogen, hydroxy, C_1-6Alkoxy, cyano, nitro, C₁-₆Alkyl, halo C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_3-10Cycloalkyl, 3-6 membered heterocyclyl containing 1-3 atoms selected from N, O and S;

x is-O-, -NH-, -CH₂–、–NH–CO–、–CO–NH–、–CO–、–CH＝CH–、-C≡C-、–CO–NH–S(O)₂–、–NH–CO–NH–、–NH–CO–O–、–OCONH–、–SO₂–、––SO₂NH–、–NHSO₂–、

Or X is absent;

l is selected from- (CH)₂)_m1–(OCH₂CH₂)_m2–、–(CH₂OCH₂)_m3–、–(CH₂)_m4–W1-(CH₂)_m5–、–(CH₂)_m4–W2-(CH₂CH₂O)_m6–(CH₂)_m7-, wherein W1 and W2 are each independently selected from-O-, -CH₂-, a five-membered heteroaromatic ring, a six-membered heteroaromatic ring, C_3-10An alkyl ring or a 3-6 membered heteroalkyl ring;

m1 is 0, 1, 2, 3,4, 5,6, 7, 8,9, 10, 11 or 12;

m2 is 0, 1, 2, 3,4, 5 or 6;

m3 is 0, 1, 2, 3,4 or 5;

m4 is 0, 1, 2, 3,4 or 5;

m5 is 0, 1, 2, 3,4, 5 or 6;

m6 is 0, 1, 2, 3 or 4;

m7 is 0, 1, 2, 3,4, 5,6 or 7;

y is-CH₂–、–CH＝CH–、-C≡C-、–O–、–NR^2a–、–CO–NR^2b–、–NR^2c–CO–CH₂O–、–NR^2d–CO–CH₂NR^2e-or Y is absent, -CO-NR^2b–、–NR^2c–CO–CH₂O–、–NR^2d–CO–CH₂NR^2eThe form of the linkage to L is respectively-L-CO-NR^2b–、–L–NR^2c–CO–CH₂O–、–L–NR^2d–CO–CH₂NR^2e-; wherein said R^2a、R^2b、R^2c、R^2d、R^2eEach independently of the others is optionally selected from hydrogen or C₁-C₄A hydrocarbyl group;

is selected from C_6-12Aryl, 5-8 membered cycloalkyl, 5-8 membered heterocyclyl or 5-8 membered heteroaryl containing 1-3 heteroatoms selected from N, O and S, 5-8 membered cycloalkyl and C_6-12Aryl radical, containing 15-8 membered heterocyclyl or 5-8 membered heteroarylo C of 3 heteroatoms selected from N, O and S_6-12An aryl group;

R₄selected from hydrogen atoms, halogens, C_1-6Alkoxy, cyano, amino, nitro, C_1-6Alkyl radical, C_3-10Cycloalkyl, 5-8 membered heterocyclyl, C_6-10Aryl, 5-6 membered heteroaryl, -NHC (O) R₅、-NHC(O)OR₆、-NR₇R₈Wherein said alkyl or alkoxy is optionally further substituted by one or more groups selected from halogen, hydroxy, C₁-C₆Alkoxy, cyano, nitro substituent;

R₅、R₆、R₇and R₈Each independently selected from hydrogen atom, C_1-6Alkyl radical, C_3-10Cycloalkyl, 5-8 membered heterocyclyl, C_6-10Aryl or 5-8 membered heteroaryl, wherein said alkyl, cycloalkyl, aryl or heteroaryl is optionally further substituted with one or more substituents selected from halogen, hydroxy, C₁-C₆Alkoxy, cyano, nitro substituent;

n is 0, 1, 2, 3, 4;

R₁is hydrogen, deuterium or CH₃；

Wherein X₁-X₇Each independently is C or N, preferably, X₁-X₇In the five-membered ring and six-membered ring and R₂、R₃And E is selected from the group consisting of:

R₂is hydrogen or substituted or unsubstituted C_1-8Hydrocarbyl, wherein said substitution is substituted with one or more substituents selected from the group consisting of: halogen, cyano, nitro, hydroxy and amino;

e is selected from-O-, -CO-NH-, -NH-CO-, -O-CH₂–、–CO–NH–CH₂–、–NR₆–、–NR₆–CH₂–、–CH₂–NR₆–、–S–、–CH(OH) -or-CH₂-; preferably E is selected from-O-or-NH-;

R₃selected from substituted or unsubstituted C_1-6Alkyl, substituted or unsubstituted C_1-6Alkoxy, substituted or unsubstituted amino, substituted or unsubstituted C_2-6Alkylcarbonyl, wherein said substitution means substitution with one or more of the following substituents: halogen, hydroxy, C_1-6Alkoxy, cyano, nitro, C_1-6Alkyl, halo C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_3-10Cycloalkyl, 3-8 membered heterocyclyl containing 1-3 atoms selected from N, O and S.

Preferably, the compound of formula (I) is selected from the following formulae:

wherein R is₁、R₂、R₃、E、R₄、n、

X, L, Y and

the definitions of (a) are the same as in the above definitions.

More preferably, the compound of formula (I) is selected from the following formulae:

wherein R is₃-E-represents a group selected from the following groups:

R₁is hydrogen, deuterium or CH₃；

R₂Is hydrogen or substituted or unsubstituted C₁-C₃Alkyl, wherein said substitution means one or more selected fromSubstituted with a variety of substituents: halogen, cyano, nitro, hydroxy and amino;

R₄selected from hydrogen atom, halogen, cyano, amino, nitro, -NHC (O) CH₃3-6 membered cycloalkyl, C_1-4Alkoxy radical, C₁-₄Alkyl, wherein said alkyl or alkoxy is optionally further substituted with one or more substituents selected from the group consisting of halogen, hydroxy, cyano, nitro;

n is 0, 1, 2;

selected from phenyl or naphthalenyl, 5-6 membered cycloalkyl, 5-6 membered heterocyclyl or 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O and S, 5-6 membered cycloalkylacenyl, 5-8 membered heterocyclyl or 5-6 membered heteroarylacenyl containing 1-3 heteroatoms selected from N, O and S;

x, L, Y and

the definitions of (a) are the same as in the above definitions.

Further preferably, the compound of formula (I) is selected from the following formulae:

wherein R is₃-E-represents a group selected from the following groups:

R₁is hydrogen, deuterium or CH₃；

R₂Is hydrogen or substituted or unsubstituted C_1-3Alkyl, wherein said substitution is substituted with one or more substituents selected from the group consisting of: halogenElements, cyano, nitro, hydroxy and amino;

R₄is hydrogen atom, halogen, cyano, amino, nitro, -NHC (O) CH₃3-6 membered cycloalkyl;

n is 0, 1, 2;

selected from the following groups:

l is selected from- (CH)₂)_m1–(OCH₂CH₂)_m2–、–(CH₂OCH₂)_m3–、–(CH₂)_m4–W1-(CH₂)_m5–、–(CH₂)_m4–W2-(CH₂CH₂O)_m6–(CH₂)_m7–；

Wherein W1 and W2 are independently and preferably selected from-O-, -CH₂-、

X、Y、

m1, m2, m3, m4, m5, m6, m7 are as defined above.

In a preferred embodiment, wherein the compound of formula (I) is selected from the following formulae:

wherein R is₁、R₂、R₃E, X, L, Y and

the definitions of (a) are the same as in the above definitions.

wherein R is₁、R₂、R₃、E、R₄、n、X、L、Y、R_c1、R_a2、R_a3And R_a4The definitions of (a) are the same as in the above definitions.

wherein R is₁、R₂、R₃、E、R₄、n、

X, L, Y and

the definitions of (a) are the same as in the above definitions.

More preferably, the compound of formula (I) is selected from the following compounds:

a tautomer, enantiomer, diastereomer, racemate, metabolic precursor, pharmaceutically acceptable salt, ester, prodrug, or hydrate thereof.

The term "halogen" refers to fluorine, chlorine, bromine or iodine.

The term "hydrocarbyl" refers to a substituent containing only carbon and hydrogen atoms, including, without limitation, methyl, ethyl, isopropyl, propyl, cyclohexyl, phenyl, and the like.

The term "C₁-C₈Alkyl "refers to a straight or branched chain saturated hydrocarbon group having 1 to 8 carbon atoms in the chain, including, without limitation, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, and the like.

The term "cycloalkyl" refers to a saturated cyclic alkyl group consisting of carbon atoms, including without limitation cyclobutyl, cyclopentyl, cyclohexyl, and the like.

The term "haloalkyl" refers to a straight, branched, or cyclic alkyl group substituted with a single or multiple halogens, including without limitation 2-bromoethyl, 2-bromopropyl, and the like.

The term "hydroxyalkyl" refers to a straight, branched or cyclic alkyl group substituted with a single or multiple hydroxyl groups, including, without limitation, (S) -1-hydroxyisobutan-2-yl, (R) -1-hydroxyisobutan-2-yl, and the like.

The term "C₅-C₈Cycloalkyl "refers to a saturated mono-or poly-ring system containing 5 to 8 carbon atoms and includes, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

The term "C₆-C₁₀Aryl "refers to aromatic ring groups containing 6-10 ring atoms, but no heteroatoms in the ring atoms, such as phenyl, naphthyl.

The term "arylene" refers to a divalent aromatic monocyclic or divalent aromatic fused bicyclic hydrocarbon group, representative examples include, but are not limited to, phenylene, naphthylene, and the like.

The term "5-10 membered heterocyclyl" means a ring containing one or more saturated and/or partially saturated rings, including 5 to 10 ring atoms, wherein one or more ring atoms are selected from heteroatoms of nitrogen, oxygen or sulfur, the remaining ring atoms being carbon; for example, propylene oxide, tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl.

The term "5-12 membered heteroaryl" refers to a monovalent aromatic ring group containing 5-12 ring atoms and 1-4 heteroatoms in the ring atoms as ring members. The heteroatoms may be selected from nitrogen, oxygen or sulfur. The heteroaryl group may be a monocyclic heteroaryl group having 5 to 7 ring atoms, or a bicyclic heteroaryl group having 7 to 12 ring atoms. The bicyclic heteroaryl group may have one ring as long as it is a heteroaromatic ring, and the other ring may be aromatic or non-aromatic, and may or may not contain a heteroatom. Examples of heteroaryl groups include, but are not limited to, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, pyridyl, pyrimidinyl, furyl, thienyl, thiazolyl, thiadiazolyl, tetrazolyl, triazolyl, isoxazolyl, indolyl, methylpyridinium thiazolyl, benzo [ d ] oxazole, benzo [ d ] imidazole, and the like. Preferably, the 5-12 membered heteroaryl is selected from the group consisting of pyridyl, pyrimidinyl, oxazolyl, thienyl, indolyl, 1, 3-dioxoisoindolyl, 1-oxoisoindolyl, imidazolyl, pyrazolyl, isoxazolyl, benzimidazolyl and furanyl.

The term "arylheterocycloalkylalkyl" refers to an aryl or heteroaryl cycloalkyl group containing oxygen, nitrogen, or sulfur, and includes, without limitation, 1, 3-benzodioxole, 2, 3-dihydrobenzofuran, indoline, and the like.

The term "5-12 membered heteroarylene" refers to a divalent aromatic ring group containing 5-12 ring atoms and 1-4 heteroatoms in the ring atoms as ring members. The heteroatoms may be selected from nitrogen, oxygen or sulfur. The heteroarylene group may be a monocyclic heteroarylene group having 5 to 7 ring atoms, or a bicyclic heteroarylene group having 7 to 12 ring atoms. For example, triazolylene, pyrrolylene, pyrazolyl, imidazolyl, oxazolylene, pyridyl, pyrimidylene, furanylene, thienyl, thiazolyl, thiadiazolylene, tetrazolylene, isoxazolylene, indolyl, methylenepyridinium thiazolyl, benzoxazolyl, benzimido, and the like.

The term "tautomer" refers to structural isomers that readily interconvert by a chemical reaction with each other being an isomer, which reaction generally results in formal movement of hydrogen atoms or protons with concomitant transformation of single bonds and adjacent double bonds.

The term "enantiomer" refers to stereoisomers that are mirror images of each other and do not overlap, such as compound C045 and its enantiomers.

"diastereomer" refers to a stereoisomer that has two or more chiral neutrals and is not a mirror image, e.g., compound C045 and its diastereomer.

"racemic" refers to two stereoisomers which are mirror images of each other, and have opposite optical rotation and the optical rotation is cancelled, for example, compound C001, compound C002, compound C003, and the like.

"metabolic precursors" refers to compounds that are inactive or less active in vitro and that release the active drug to exert their pharmacological effects in vivo through the conversion of metabolic pathways in vivo.

"pharmaceutically acceptable salts" refers to salts formed by a drug molecule and corresponding organic acid, inorganic acid or organic and inorganic bases, such as hydrochloric acid, formic acid, trifluoroacetic acid, succinic acid, methanesulfonic acid salt of compound C001.

"prodrug" refers to a class of compounds that are inactive or less active in vitro and exert their pharmaceutical effects in vivo by converting enzymatically or non-enzymatically to release the active drug.

"hydrate" refers to a compound containing water.

The present invention also encompasses any of the novel intermediates disclosed herein.

In one aspect, the present invention provides a method for preparing a degradation agent represented by the general formula (I), wherein the method is selected from one of the following methods:

the first synthesis method comprises the following steps:

synthetic references to ligands targeting CDK targets (1A) j.med.chem.2016,59, 8667-8684; bioorganic & Medicinal chemistry letters2015, 25, 3420-; med, chem, 2009, 52, 655-; med, chem, 2010,53, 8508-; and US2007179161a1, US2015111873a1, WO2016193939a1, WO2016142855a 2.

Wherein R is₂、R₃、E、X₁、X₂And X₇The definition of (a) is the same as above;

n₁n is an integer of 0 to 12₂Is an integer of 0 to 6;

step a, carrying out condensation reaction on a compound 1A and a connecting chain 1B to obtain a compound 1C;

step b Compound 1C with 1D in diisopropyl azodicarboxylate (DIAD) and triphenylphosphine (PPh)₃) Reaction under conditions gave 1E.

And a second synthesis method comprises the following steps:

u is-O-or-CH₂-；

n₃Is an integer of 0 to 9, n₄Is an integer of 0 to 5;

step C, reacting the compound 2A with the connecting chain 2B under the conditions of cuprous iodide and bis (triphenylphosphine) palladium dichloride to obtain a compound 2C;

d, reacting the compound 2C under the conditions of hydrogen and a palladium-carbon hydrogenation catalyst to obtain a compound 2D;

and E, carrying out condensation reaction on the compound 2D and the compound 1A to obtain a compound 2E.

The third synthesis method comprises the following steps:

n₅is an integer of 0 to 12, n₆Is an integer of 0 to 6;

step f, carrying out condensation reaction on the connecting chain 3A and the compound 1A to obtain a compound 3B;

step g, the compound 3B obtains a compound 3C under the deprotection condition of hydrochloric acid or trifluoroacetic acid;

step h, carrying out condensation reaction on the compound 3C and the compound 3D to obtain a compound 3E;

step i, reacting the compound 3C with the compound 3F under the condition of N, N-diisopropylethylamine to obtain a compound 3G.

The synthesis method comprises the following steps:

z is methylene or carbonyl;

n₇is an integer of 0 to 12, n₈Is an integer of 0 to 6;

step j Compounds 4A and 4B in DIAD and PPh₃Reacting under the condition to obtain 4C;

step k, reacting the compound 4C under the condition of tetrabutylammonium fluoride to obtain a compound 4D;

step l, reacting the compound 4E in the presence of hydrogen and a palladium-carbon hydrogenation catalyst to obtain a compound 4F;

and m, carrying out condensation reaction on the compound 1A and the connecting chain 4F to obtain a compound 4G.

The synthesis method comprises the following steps:

n₉is an integer of 0 to 10, n₁₀Is an integer of 0 to 6;

step n, carrying out condensation reaction on the compound 5A and the connecting chain 5B to obtain a compound 5C;

step o, removing a protecting group of the compound 5C under the condition of lithium hydroxide to obtain a compound 5D;

step p: the compound 5D and the compound 1A are subjected to condensation reaction to obtain a compound 5E.

The synthesis method comprises the following steps:

wherein R is₂、R₃、Ra3、Ra4、E、X₁、X₂And X₇The definition of (a) is the same as above;

n₁₁is an integer of 0 to 6, n₁₂Is an integer of 0 to 10;

step q, carrying out condensation reaction on the compound 6A and the connecting chain 6B to obtain a compound 6C;

step r, removing a protecting group of the compound 6C under the condition of lithium hydroxide to obtain a compound 6D;

step s: the compound 6D and the compound 1A are subjected to condensation reaction to obtain a compound 6E.

The synthesis method comprises the following steps:

n₁₃is an integer of 0 to 9, n₁₄Is an integer of 0 to 5;

step u: reacting the compound 2A with the connecting chain 7A under the conditions of cuprous iodide and bis (triphenylphosphine) palladium dichloride to obtain a compound 7B;

step v: reacting the compound 7B under the condition of trifluoroacetic acid to obtain a compound 7C;

step w: condensing the compound 7C and the compound 1A under the conditions of HATU and DIPEA to obtain a compound 7D;

step x: the compound 2C is catalyzed by palladium carbon to obtain a compound 7E under the condition of normal pressure hydrogen.

The synthesis method comprises the following steps:

n₁₅is an integer of 0 to 9, n₁₆Is an integer of 0 to 5;

8F Synthesis references J.Med.chem.2016,59, 8667-8684; bioorganic & medicinal chemistry Letters2015, 25, 3420-; med, chem, 2009, 52, 655-; med, chem, 2010,53, 8508-; and US2007179161a1, US2015111873a1, WO2016193939a1, WO2016142855a 2.

Step y: reacting the compound 8A and the compound 8B under the condition of bis (triphenylphosphine) palladium dichloride and cuprous iodide to obtain a compound 8C;

step z 1: hydrogenating the compound 8C under the catalysis of Raney nickel to obtain a compound 8D;

step z 2: reacting the compound 8D under the conditions of imidazole and TBSCl to obtain a compound 8E;

step z 3: carrying out nucleophilic substitution reaction on a compound 8E and a compound 8F under an alkaline condition to prepare a compound 8G;

step z 4: compounds 8G and R₃EH is subjected to nucleophilic substitution reaction under the alkaline condition to prepare a compound 8H;

step z 5: reacting the compound 8H under the condition of tetrabutylammonium fluoride to obtain a compound 8I;

step z 6: the compound 8I reacts under the conditions of diisopropyl azodicarboxylate and triphenylphosphine to obtain a compound 8J.

The invention provides a compound with a structure shown as a formula (I) or a crystal hydrate and a solvate, and application of the compound or the crystal hydrate and the solvate in preparation of a medicament for preventing and/or treating diseases or symptoms related to abnormal activity of CDK.

When the compound contains basic groups, the compound can be prepared into pharmaceutically acceptable salts, including inorganic acid salts and organic acid salts. Suitable acids for forming the salts include (but are not limited to): inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, etc., organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, phenylmethanesulfonic acid, benzenesulfonic acid, etc.; and acidic amino acids such as aspartic acid and glutamic acid.

In yet another aspect, the present invention provides a pharmaceutical composition, which comprises a therapeutically effective amount of a compound represented by the general formula (I), and one or more of stereoisomers, pharmaceutically acceptable salts, prodrugs, solvates, hydrates, and crystal forms thereof, and at least one excipient, diluent, or carrier.

Typical formulations are prepared by mixing a compound of formula (I) of the invention with a carrier, diluent or excipient. Suitable carriers, diluents or excipients are well known to 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 excipient employed will depend upon the mode of use and the purpose of the compound of the invention. The solvent is generally selected based on the solvent that one of skill in the art would consider safe and effective for administration to mammals. Generally, safe solvents are non-toxic aqueous solvents such as water, as well as other non-toxic solvents that are soluble or miscible with water. Suitable aqueous solvents include one or more of water, ethanol, propylene glycol, polyethylene glycol (e.g., PEG400, PEG300), and the like. The formulation may also include one or more buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, glidants, processing aids, colorants, sweeteners, flavoring agents or other known additives to make or use the drug in an acceptable form.

When the compound of formula (I) according to the present invention is used in combination with at least one other drug, the two drugs or more may be used separately or in combination, preferably in the form of a pharmaceutical composition. The compounds or pharmaceutical compositions of the invention according to formula (I) can be administered to a subject separately or together in any known oral, intravenous, rectal, vaginal, transdermal, other topical or systemic administration form.

These pharmaceutical compositions may also contain one or more buffering agents, stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifying agents, suspending agents, preservatives, antioxidants, opacifiers, glidants, processing aids, colorants, sweeteners, flavoring agents or other known additives to allow the pharmaceutical composition to be manufactured or used in an acceptable form.

The route of oral administration is preferred for the medicaments of the invention. Solid dosage forms for oral administration may include capsules, tablets, powders or granules. In solid dosage forms, the compounds or pharmaceutical compositions of the present invention are mixed with at least one inert excipient, diluent or carrier. Suitable excipients, diluents or carriers include substances such as sodium citrate or dicalcium phosphate, or starches, lactose, sucrose, mannitol, silicic acid and the like; binders such as carboxymethyl cellulose, alginate, gelatin, polyvinyl pyrrolidone, sucrose, gum arabic, etc.; humectants such as glycerin, etc.; disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, specific complex silicate, sodium carbonate, etc.; solution retarding agents such as paraffin, etc.; absorption accelerators such as quaternary ammonium compounds and the like; adsorbents such as kaolin, bentonite, etc.; lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and the like. In the case of capsules and tablets, the dosage form may also include buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using lactose and high molecular weight polyethylene glycols and the like as excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs. In addition to the compounds of the present invention or compositions thereof, the liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents; solubilizers and emulsifiers such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3-butylene glycol, dimethylformamide; oils (e.g., cottonseed oil, groundnut oil, corn germ oil, olive oil, castor oil, sesame oil, etc.); glycerol; tetrahydrofurfuryl alcohol; fatty acid esters of polyethylene glycol and sorbitan; or mixtures of several of these substances, and the like.

In addition to these inert diluents, the compositions can also include excipients such as one or more of wetting agents, emulsifying agents, suspending agents, sweetening, flavoring, and perfuming agents.

For suspensions, in addition to the compounds or combinations of the present invention, carriers such as suspending agents, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol, sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, tragacanth, or mixtures of several of these, may be included.

The compounds or pharmaceutical compositions of the present invention may be administered in other topical dosage forms including creams, powders, sprays and inhalants. The medicament may be mixed under sterile conditions with a pharmaceutically acceptable excipient, diluent or carrier, and any preservatives, buffers or propellants which may be required. Ophthalmic formulations, ophthalmic ointments, powders and solutions are also intended to be within the scope of the present invention.

In yet another aspect, the present invention provides the use of a compound of formula (i) as well as tautomers, enantiomers, diastereomers, racemates, metabolic precursors, pharmaceutically acceptable salts, esters, prodrugs or hydrates thereof, or pharmaceutical compositions thereof, for the manufacture of a medicament for the prevention and/or treatment of diseases or conditions associated with abnormal activity of CDKs.

The diseases related to the abnormal activity of CDKs comprise solid tumors such as breast cancer, colon cancer, prostatic cancer, small cell lung cancer, non-small cell lung cancer and the like, and blood tumors such as acute lymphocytic leukemia, B cell lymphoma, T cell lymphoma, myeloma, acute and chronic myelocytic leukemia, promyelocytic leukemia and the like.

The invention also provides the use of the compound shown in the general formula (I), and a stereoisomer, a pharmaceutically acceptable salt, a prodrug, a solvate, a hydrate or a crystal form thereof or the pharmaceutical composition in the preparation of medicaments for treating diseases such as tumors, central system diseases, immune diseases and the like.

In a preferred embodiment, the disease includes, but is not limited to, cancer, angiogenesis-related diseases or disorders, pain (including but not limited to complex regional pain syndrome), macular degeneration and related disorders, skin diseases, pulmonary disorders, immunodeficiency disorders, central nervous system injuries and disorders, and TNF α -related diseases or disorders.

In another preferred embodiment, the cancer includes (but is not limited to): skin cancer (such as melanoma), cancer of the lymphatic system, breast cancer, cervical cancer, uterine cancer, cancer of the digestive tract, lung cancer, ovarian cancer, prostate cancer, colon cancer, rectal cancer, oral cancer, brain tumor, head and neck cancer, cancer of the throat, cancer of the testis, kidney cancer, pancreatic cancer, spleen cancer, liver cancer, bladder cancer, cancer of the larynx, and cancer associated with AIDS. The compounds provided by the invention are also effective against hematological neoplasms and myelomas, such as can be used to treat multiple myeloma and acute and chronic leukemia. The compound provided by the invention can be used for preventing or treating primary tumor and metastatic tumor.

It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments.

Drawings

FIG. 1 shows the effect of Western Blot on CDK degradation by compounds C022 and C025

Detailed Description

The present invention is further illustrated below with reference to examples, which are by no means intended to limit the scope of the invention.

In all of the embodiments described herein, the first,¹h NMR was recorded by a Bruker Avance model III-300 or Avance model III-400 nuclear magnetic resonance apparatus, chemical shifts are expressed in delta (ppm); mass spectra were determined by MS mass spectra UPLC-MS (esi); wherein the UPLC model is Waters HPLC H-CLASS, and the MS (ESI) model is Waters SQ Detector 2; the anhydrous tetrahydrofuran is prepared by benzophenone/metallic sodium reflux drying and deoxidization, and the anhydrous toluene and the anhydrous dichloromethane are prepared by calcium chloride reflux drying; solvents such as petroleum ether, ethyl acetate and dichloromethane for column chromatography mobile phase are all purchased from chemical reagents of national medicine group; the thin layer chromatography silica gel plate (HSGF254) used in the reaction detection is from chemical reagents of national drug group, Inc.; the compound separation is performed by using 200-300 mesh silica gel of national drug group chemical reagent, Inc. Reagents such as o-nitrobenzenesulfonyl chloride, thioglycolic acid, diglycolamine and the like are purchased from chemical reagents of national drug group, ltd.

EtOH: ethanol; DCM: dichloromethane; TFA: trifluoroacetic acid; MeOH: methanol; NaOH: sodium hydroxide; HCl: hydrogen chloride; TEA: triethylamine; 1, 4-dioxane: 1, 4-dioxane; NaH: sodium hydride; h₂O: water; HATU: 2- (7-benzotriazole oxide) -N, N, N ', N' -tetraMethyl urea hexafluorophosphate; DMF: n, N-dimethylformamide; THF: tetrahydrofuran; DIPEA: n, N-diisopropylethylamine; AcOH: acetic acid; k₂CO₃: potassium carbonate; cs₂CO₃: cesium carbonate; pd (dppf) Cl₂: [1, 1' -bis (diphenylphosphino) ferrocene]Palladium dichloride; LiOH: lithium hydroxide; PPh₃: triphenylphosphine; DEAD: diethyl azodicarboxylate; the DIAD: diisopropyl azodicarboxylate; KOH: potassium hydroxide; MsCl: methanesulfonyl chloride; NaN₃: sodium azide; Pd/C: a palladium on carbon hydrogenation catalyst; pd₂dba₃: tris (dibenzylideneacetone) dipalladium; rac-BINAP: 1,1 '-binaphthyl-2, 2' -bis-diphenylphosphine; toluene: toluene; t-BuONa: sodium tert-butoxide; (Boc)₂O: di-tert-butyl dicarbonate; and (2) CuI: cuprous iodide; pd (PPh)₃)₂Cl₂: bis (triphenylphosphine) palladium dichloride; EA: ethyl acetate; DMAP: 4-dimethylaminopyridine; NaHCO 2₃: sodium bicarbonate; KI: potassium iodide; HCOOH: formic acid; Fmoc-OSU: 9-fluorenylmethyl-N-succinimidyl carbonate; NH (NH)₄Cl: ammonium chloride; piperidine: piperidine; POCl₃: phosphorus oxychloride; PhNMe₂: n, N-dimethylaniline; MeCN: acetonitrile; raney Ni: raney nickel; BPO: benzoyl peroxide; NBS: n-bromosuccinimide; benzene: benzene; p-TsCl: 4-methylbenzenesulfonyl chloride; pyridine: pyridine; DMSO, DMSO: dimethyl sulfoxide; NaBH₃CN: sodium cyanoborohydride; TBSCl: tert-butyldimethylsilyl chloride; imidazole: imidazole; CBZ-Cl: benzyl chloroformate; m-CPBA: m-chloroperoxybenzoic acid; DBU: 1, 8-diazabicyclo [5.4.0 ]]Undec-7-ene; BH₃THF: borane tetrahydrofuran; NMP: n-methyl pyrrolidone.

First, preparation example

Example 1.

[C001] Preparation of N- (6- (4- (((5- ((6-N-hexylamino) amino) -3-isopropylpyrazolo [1,5-a ] pyrimidin-7-yl) amino) methyl) phenyl) N-hexyl) -2- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) acetamide

P-bromobenzylamine (1g, 5.37mmol) was dissolved in methylene chloride, triethylamine (1.23mL, 5.37mmol, 1equiv) was added, di-tert-butyl dicarbonate was added dropwise at 0 ℃ to react at room temperature for 12 hours, and methylene chloride and saturated NaHCO were added to the reaction solution₃Washing with anhydrous Na₂SO₄Drying and column chromatography gave 1.1(1.35g) with 88% yield.¹H NMR(400MHz,CDCl₃)δ7.43(d,J＝7.5Hz,2H),7.15(d,J＝8.0Hz,2H),4.25(d,J＝5.6Hz,2H),1.45(s,9H)。

Dissolving 6-chlorohexyne (1g, 8.58mmol) and potassium phthalimide (2.54g, 13.72mmol, 1.6equiv) in 10mL DMF, reacting at 70 deg.C for 18 hr, adding water to the reaction solution, extracting with ethyl acetate, mixing ethyl acetate layers, washing with saturated NaCl, and adding anhydrous Na₂SO₄Drying and column chromatography gave 1.2(1.82g) with 85% yield.¹H NMR(400MHz,CDCl₃)δ7.83(dd,J＝5.4,3.1Hz,2H),7.70(dd,J＝5.4,3.1Hz,2H),3.70(t,J＝7.1Hz,2H),2.23(td,J＝7.0,2.6Hz,2H),1.93(t,J＝2.6Hz,1H),1.84-1.75(m,2H),1.61-1.51(m,2H)。

Compound 1.1(286mg, 1mmol), compound 1.2(454mg, 2mmol, 2equiv), cuprous iodide (38mg, 0.2mmol, 20 mol%), bis-triphenylphosphine palladium dichloride (70mg, 0.1mmol, 10 mol%) were added to the reaction flask, N₂Protection, 2.5mL of triethylamine and 5mL of DMMF, N are added₂Protecting, reacting at 80 ℃ for 12 hours, filtering the reaction solution through kieselguhr, washing with 120mL ethyl acetate, washing with water for several times, washing with saturated NaCl, and washing with anhydrous Na₂SO₄Drying and column chromatography gave 1.3(405mg) in 89% yield.¹H NMR(400MHz,CDCl₃)δ7.84(dd,J＝5.4,3.1Hz,2H),7.71(dd,J＝5.4,3.0Hz,2H),7.33(d,J＝8.2Hz,2H),7.18(d,J＝8.1Hz,1H),4.28(d,J＝5.7Hz,1H),3.74(t,J＝7.1Hz,2H),2.46(t,J＝7.0Hz,2H),1.91-1.81(m,2H),1.69-1.60(m,2H),1.45(s,9H)。

Dissolving compound 1.3(405mg, 0.94mmol) in 25mL ethyl acetate and 25mL methanol, adding palladium carbon (content 5%, 81mg), introducing 3atm hydrogen, reacting at room temperature for 6 hr, filtering off palladium carbon, and removing solvent under reduced pressure to obtain the final productObject 1.4.¹H NMR(400MHz,CDCl₃)δ7.83(dd,J＝5.4,3.1Hz,2H),7.70(dd,J＝5.3,3.1Hz,2H),7.17(d,J＝7.9Hz,2H),7.10(d,J＝8.0Hz,2H),4.26(d,J＝5.5Hz,2H),3.66(t,J＝7.3Hz,2H),2.56(t,J＝7.7Hz,2H),1.70-1.53(m,4H),1.45(s,9H),1.39-1.26(m,4H)。

Compound 1.5(76mg, 0.33mmol) (see EP2634190a1), compound 1.4(165mg, 0.5mmol, 1.5equiv) were dissolved in 10mL of ethanol, DIPEA (109 μ L, 0.66mmol, 2equiv) was added, reflux was carried out for 12 hours, the solvent was removed under reduced pressure, and column chromatography was carried out to give product 1.6(69mg) with a yield of 39%.¹H NMR(400MHz,CDCl₃)δ7.83-7.78(m,3H),7.71-7.64(m,2H),7.34-7.18(m,2H),7.11(dd,J＝26.2,7.1Hz,2H),5.90(d,J＝1.0Hz,1H),4.49(d,J＝5.5Hz,2H),3.65(t,J＝7.3Hz,2H),3.29-3.19(m,1H),2.57(t,J＝7.5Hz,2H),1.69-1.51(m,4H),1.39-1.27(m,10H)。

Compound 1.6(69mg, 0.13mmol) was dissolved in 3mL THF and DMAP (3mg, 0.026mmol, 0.2equiv) and (Boc) were added, respectively, at room temperature₂O (46. mu.L, 0.2mmol, 1.5equiv), reacted at room temperature for 12 hours, the solvent was removed under reduced pressure, and column chromatography was performed to obtain 1.7(78mg) as a product with a yield of 95%.

Compound 1.7(78mg, 0.124mmol), Pd₂dba₃(6mg，0.0062mmol，5mol％)、rac-BINAP(12mg，0.0186mmol，15mol％)、Cs₂CO₃(60mg, 0.186mmol, 1.5equiv) in 3mL of anhydrous toluene, N₂After reaction at room temperature for 15 minutes under protection, Compound 1.8(433mg, 2mmol, 2equiv) was dissolved in 2mL of anhydrous toluene, added to the reaction mixture, and then purged again, N₂Protecting, reacting at 100 deg.C for 16 hr, adding ethyl acetate, washing with water and saturated NaCl, and adding anhydrous Na₂SO₄Drying and column chromatography gave 1.9(40mg) in 40% yield.

Compound 1.9(40mg, 0.0494mmol) was dissolved in 4mL of ethanol, hydrazine hydrate (85%, 57 μ L, 0.988mmol, 20equiv) was added, and after refluxing for 4 hours, the solid therein was filtered off, washed with ethyl acetate, the solvent was removed under reduced pressure, and column chromatography was performed to give product 1.10(22mg) with a yield of 65%.¹H NMR(400MHz,CDCl₃)δ7.72(s,1H),7.14(d,J＝8.1Hz,2H),7.08(d,J＝8.1Hz,2H),566(s,1H),4.89(d, J ═ 5.7Hz,2H),3.31(dd, J ═ 12.7,6.7Hz,2H),3.11(dt, J ═ 13.7,6.9Hz,3H),2.67(t, J ═ 7.0Hz,2H),2.59-2.52(m,2H),1.63-1.51(m,4H),1.50-1.26(m,36H), UPLC-ms (esi) theoretical values are C, 1H, d, J ═ 5.7Hz,2H, d, J ═ 13.7Hz, 3.7H, d₃₈H₆₁N₇O₄[M+H]⁺680.48, found 680.87.

Compound 1.10(22mg, 0.0324mmol), Compound 1.11(10.7mg, 0.0324mmol, 1equiv) (see Lohbeck J., Miller A. K. practical Synthesis of a phenyl-based Cereblock ligand to an enabled PROTAC reduction. bioorganic for preparation of Compound 1.11&medicinalcohemistry letters.2016; 5260 (5260) -₂SO₄Drying, filtering, removing solvent under reduced pressure, and performing column chromatography to obtain product 1.12(30mg) with yield of 94%.

Compound 1.12(30mg, 0.0302mmol) was dissolved in 3mL of dichloromethane, 800. mu.L of trifluoroacetic acid was added dropwise at 0 ℃ and reacted at room temperature for 3 hours, the solvent was removed under reduced pressure and purified by HPLC to give C001(21mg) in 99% yield.¹H NMR(400MHz,DMSO-d₆) δ 11.13(s,1H),7.95(t, J ═ 5.6Hz,1H),7.78(dd, J ═ 8.5,7.3Hz,1H),7.48(d, J ═ 7.2Hz,1H),7.37(d, J ═ 8.6Hz,1H),7.29(d, J ═ 8.0Hz,2H),7.15(d, J ═ 8.1Hz,2H),5.32(s,1H),5.11(dd, J ═ 12.9,5.4Hz,1H),4.76(s,2H),4.56(s,2H),3.32-3.22(m,2H),3.17-3.00(m,3H),2.94-2.82 (esim, 1H),2.80-2.71 (s,2H), 2.32-3.22 (m,2H), 1.17-3.00 (m,3H),2.94-2.82 (esi, 1H), 2.71(m, 2.71-2H), 1.56 (m, 14.6H), 1H, 14 (lc-1H), 1H, 14 (lc-36 (H), 1H₄₃H₅₅N₉O₆[M+H]⁺794.43, found 794.81.

[C002] Preparation of N- (6- (3- (((5- ((6-amino-N-hexyl) amino) -3-isopropylpyrazolo [1,5-a ] pyrimidin-7-yl) amino) methyl) phenyl) N-hexyl) -2- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxo) acetamide

C002 was prepared by substituting p-bromobenzylamine with m-bromobenzylamine according to the synthetic route of example 1.¹H NMR(400MHz,DMSO-d₆) δ 11.13(s,1H),7.98(s,1H),7.78(dd, J ═ 8.4,7.4Hz,1H),7.48(d, J ═ 7.2Hz,1H),7.38(d, J ═ 8.5Hz,1H),7.28-7.17(m,3H),7.09(d, J ═ 7.2Hz,1H),5.39(s,1H),5.11(dd, J ═ 12.9,5.4Hz,1H),4.77(s,2H),4.63(s,2H),3.31-3.22(m,2H),3.18-3.01(m,3H),2.94-2.81(m,1H),2.79-2.71(m,2H),2.63-2.44(m, 4.44(m, 1H), 1.18-3.58 (m,1H), theoretical values (lc, 1H), 1H-36 (1H), 1H, 13-36 (lc, 13H), 1H, 1₄₃H₅₅N₉O₆[M+H]⁺794.43, found 794.92.

Example 2

[C012] Preparation of 7- (2- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) acetamido) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) heptanamide

4-Nitro benzylamine hydrochloride (590mg, 3.13mmol) dissolved in 15mL of dichloromethane, DIPEA (1.03mL, 6.26mmol, 2equiv) and Fmoc-OSU (1.1g, 3.28mmol, 1.05equiv) dissolved in dichloromethane at 0 ℃ were added dropwise to the reaction mixture, the mixture was reacted at room temperature for 4 hours, 100mL of dichloromethane was added to the reaction, 0.5N aqueous HCl was washed several times, saturated NaCl was washed, anhydrous Na₂SO₄Drying, filtering, removing solvent under reduced pressure, and performing column chromatography to obtain product 2.1(1.15g) with yield of 98%.¹H NMR(400MHz,CDCl₃)δ8.18(d,J＝8.5Hz,2H),7.78(d,J＝7.6Hz,2H),7.59(d,J＝7.4Hz,2H),7.46-7.29(m,6H),4.54(d,J＝6.4Hz,2H),4.45(d,J＝6.3Hz,2H),4.22(t,J＝6.4Hz,1H).

Compound 2.1(600mg, 1.6mmol) in THF/MeOH/H₂Adding zinc powder (1.05g, 16mmol, 10equiv) and ammonium chloride (856mg, 16mmol, 10equiv) into O (6mL, 4mL, 2mL) mixed solvent, reacting at 50 deg.C for 4 hr, filtering the reaction solution with diatomaceous earth, washing with dichloromethane, removing solvent under reduced pressure, dissolving with dichloromethane again, washing with water for several times, and adding anhydrous Na₂SO₄Drying, filtering, and removing solvent under reduced pressureThis gave product 2.2(531 mg).¹H NMR(400MHz,DMSO-d₆)δ7.89(d,J＝7.4Hz,2H),7.69(d,J＝7.5Hz,2H),7.41(t,J＝7.2Hz,2H),7.32(t,J＝7.5Hz,2H),6.88(d,J＝8.3Hz,2H),6.49(d,J＝8.3Hz,2H),4.30(d,J＝7.1Hz,2H),4.21(t,J＝7.0Hz,1H),4.00(d,J＝6.0Hz,2H).

The compound 2.2(950mg, 2.76mmol) was dissolved in 20mL of anhydrous dichloromethane, triethylamine (714. mu.L, 5.52mmol, 2equiv) and di-tert-butyl dicarbonate (1.3mL, 5.52mmol, 2equiv) were added to react at room temperature for 24 hours, the solvent was removed under reduced pressure, and column chromatography was performed to obtain 2.3(620mg) as a product with a yield of 51%.¹H NMR(400MHz,CDCl₃)δ7.76(d,J＝7.5Hz,2H),7.59(d,J＝7.2Hz,2H),7.40(t,J＝7.4Hz,2H),7.35-7.27(m,4H),7.19(d,J＝8.2Hz,2H),4.45(d,J＝7.0Hz,2H),4.32(d,J＝5.8Hz,2H),4.23(t,J＝6.8Hz,1H),1.52(s,9H)。

The compound 2.3(254mg, 0.57mmol) was dissolved in 5mL of dichloromethane, 1mL of piperidine was added, reaction was carried out at room temperature for 2 hours, the solvent was removed under reduced pressure, and column chromatography was carried out to give the product 2.4(125mg) with a yield of 99%.¹H NMR(400MHz,CDCl₃)δ7.32(d,J＝8.2Hz,2H),7.22(d,J＝8.5Hz,2H),3.80(s,2H),1.51(s,9H)。

Compound 2.5(297mg, 1.16mmol) (see EP2634190A1), N, N-dimethylaniline (440. mu.L, 3.47mmol, 3equiv) were dissolved in 8mL of phosphorus oxychloride and reacted at 100 ℃ for 4 hours, after which the phosphorus oxychloride was removed under reduced pressure, the residue was dissolved in acetonitrile and the acetonitrile was removed under reduced pressure, and the reaction was repeated several times to remove all of the phosphorus oxychloride. Compound 2.4(190mg, 1.16mmol, 1equiv) was dissolved in 10mL acetonitrile, DIPEA (573. mu.L, 3.47mmol, 3equiv) was added, the residue obtained in the previous step was dissolved in acetonitrile, the mixture was added to the reaction mixture, reacted at room temperature for 12 hours, acetonitrile was removed under reduced pressure, dissolved in ethyl acetate, and saturated NaHCO was added₃Washing with anhydrous Na₂SO₄Drying and column chromatography gave 2.6(330mg) in 62% yield.

Dissolving N-methyl-4-piperidinol (980mg, 8.14mmol, 3equiv) in 10mL of anhydrous DMF, adding NaH (60% dispersed in liquid paraffin, 326mg, 8.14mmol, 3equiv), adding dropwise a DMF solution of compound 2.6(1.25g, 2.7mmol) at 0 deg.C, reacting at 60 deg.C for 3 hours, adding water to the reaction solution, and extracting with ethyl acetateCollecting, mixing ethyl acetate layers, washing with saturated NaCl, and removing anhydrous Na₂SO₄Drying and column chromatography gave 2.7(900mg) in 67% yield.¹H NMR(400MHz,CDCl₃) δ 7.67(s,1H),7.31(d, J ═ 8.3Hz,2H),7.22(d, J ═ 8.5Hz,2H),5.10 to 5.00(m,1H),4.70(d, J ═ 5.8Hz,2H),3.11 to 3.02(m,1H),2.87 to 2.76(m,2H),2.38 to 2.25(m,5H),2.18 to 2.08(m,2H),1.99 to 1.90(m,2H),1.49(s,9H),1.28(d, J ═ 6.9Hz,6H), UPLC-ms (esi) theoretical values are C₂₆H₃₇N₇O₃[M+H]⁺496.30, found 496.96.

Compound 2.8 was prepared by synthesis of compound C001 using compound 2.7 as a starting material.

7-aminoheptanoic acid (1.45g, 10mmol) was dissolved in a mixed solvent of 1N aqueous NaOH (15mL, 1.5equiv) and 1,4-dioxane (15mL), di-tert-butyl dicarbonate (2.76mL, 12mmol, 1.2equiv) was slowly added at 0 ℃ to react at room temperature for 12 hours, 1,4-dioxane was removed under reduced pressure, 20mL of water was added, ether was washed, the aqueous layer was adjusted to pH 3 with 1N aqueous HCl solution, ethyl acetate was extracted three times, the ethyl acetate layers were combined, washed with saturated NaCl, anhydrous Na₂SO₄Drying, filtering and removing the solvent under reduced pressure gave the product 2.9(2.5g) in 99% yield.¹H NMR(400MHz,DMSO-d₆)δ11.98(s,1H),2.88(dd,J＝13.0,6.7Hz,2H),2.18(t,J＝7.4Hz,2H),1.54-1.41(m,2H),1.42-1.29(m,11H),1.28-1.16(m,4H)。

Compound 2.10 was prepared by synthesizing compound 1.12 from compound 2.8 and compound 2.9 as starting materials.¹HNMR(400MHz,CDCl₃) δ 7.72(s,1H),7.54(d, J ═ 8.3Hz,2H),7.26(d, J ═ 8.3Hz,2H),5.18 to 5.09(m,1H),4.73(d, J ═ 5.8Hz,2H),3.14 to 3.02(m,3H),3.01 to 2.91(m,2H),2.72 to 2.56(m,2H),2.48(s,3H),2.34(t, J ═ 7.3Hz,2H),2.28 to 2.15(m,2H),2.09 to 1.97(m,2H),1.77 to 1.65(m,2H),1.51 to 1.40(m,11H),1.38 to 1.24(m,10H), esi (m-ms-esi), theoretical value of C (C, C)₃₃H₅₀N₈O₄[M+H]⁺623.40, found 623.84.

C012 was prepared by following the synthetic route in example 1 using compound 2.10 as a starting material.¹H NMR(400MHz,DMSO-d₆)δ11.12(s,1H),7.95(d,J＝0.8Hz,1H),7.80(dd, J ═ 8.5,7.4Hz,1H),7.52(d, J ═ 8.3Hz,2H),7.49(d, J ═ 7.2Hz,1H),7.38(d, J ═ 8.5Hz,1H),7.26(d, J ═ 8.5Hz,2H),5.25-5.20(m,0.5H),5.12(dd, J ═ 12.9,5.3Hz,1H),5.07-4.98(m,0.5H),4.76(s,2H),4.59(dd, J ═ 9.6,6.4Hz,2H),3.49(d, J ═ 13.9Hz,1H),3.33(d, J ═ 11.3, 1H), 3.81 (dd, J ═ 9.6,6.4Hz,2H),3.49 (dd, 3.9.9, 1H),3.33(d, J ═ 3.7.14H, 3.7.14H), 14H, 14.14H, 14H, 14.14 (dd, 14H, 1H, 2H, 1H, 14 (dd, 2H), 7.9, 14H, 2H), 2H) 1.82-1.70(m,1H),1.64-1.49(m,2H),1.47-1.38(m,2H),1.33-1.17(m,10H). UPLC-MS (ESI) has a theoretical value of C₄₃H₅₂N₁₀O₈[M+H]⁺837.40, found 837.91.

[C013] Preparation of 7- (2- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxo) acetamido) -N- (3- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) heptanamide

Referring to the synthetic route in example 2, C013 was prepared by replacing p-nitrobenzylamine hydrochloride with m-nitrobenzylamine hydrochloride.¹H NMR(400MHz,DMSO-d₆) δ 11.12(s,1H),7.97(s,1H),7.80(dd, J ═ 8.4,7.4Hz,1H),7.56(d, J ═ 5.3Hz,1H),7.50-7.45(d, J ═ 7.3Hz,2H),7.38(d, J ═ 8.5Hz,1H),7.23(t, J ═ 7.9Hz,1H),7.00(d, J ═ 7.7Hz,1H),5.24-5.19(m,0.5H),5.12(dd, J ═ 12.9,5.4Hz,1H),5.06-4.97(m,0.5H),4.76(s,2H),4.62(dd, J ═ 10.4,6.4, 3.3H, 2(s, 3.3H), 3.3.3, 3H), 3.3.3.3H, 3.3.3 (d, 3.3H), 3.3.3.3H, 3.3.3 (d, 3H, 3.3H, 3H, 3.3.3H, 3H, 3H) 2.17(d, J ═ 15.1Hz,1H),2.07-1.94(m,2H),1.83-1.68(m,1H),1.60-1.49(m,2H),1.47-1.38(m,2H),1.32-1.20(m,10H), UPLC-ms (esi) theoretical values C₄₃H₅₂N₁₀O₈[M+H]⁺837.40, found 837.87.

Example 3

[C014] N- (4- (dimethylamino) -3- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) -7- (2- ((2- (2, 6-dioxo-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) acetamido) heptanamide

2-fluoro-5-nitrobenzonitrile (2g, 12.04mmol), dimethylamine hydrochloride (1.96g, 24.08mmol, 2equiv), potassium carbonate (5g, 36.12mmol, 3equiv) were dissolved in DMF, reacted at room temperature for 12 hours, added with 150mL ethyl acetate, washed several times with water, washed with saturated NaCl, anhydrous Na₂SO₄Drying, filtration and removal of the solvent under reduced pressure gave the product 3.1(2.24g) in 97% yield.¹HNMR(400MHz,CDCl₃)δ8.38(d,J＝2.8Hz,1H),8.15(dd,J＝9.6,2.8Hz,1H),6.78(d,J＝9.6Hz,1H),3.32(s,6H)。

Compound 3.1(2.24g, 11.72mmol) is dissolved in EtOH (20mL)/H₂O (10mL)/THF (20mL), iron powder (3.27g, 58.58mmol, 5equiv) and NH were added₄Cl (3.13g, 58.58mmol, 5equiv), refluxing for 3 hours, filtering the reaction solution through celite, washing with ethyl acetate, washing the filtrate with water and saturated NaCl, drying over anhydrous Na2SO4, filtering, removing the solvent under reduced pressure, and performing column chromatography to obtain the product 3.2(1.25g), with a yield of 66%.

Compound 3.3 was prepared by synthesizing compound 1.12 from compound 2.9 and compound 3.2 as starting materials.¹HNMR(400MHz,CDCl₃)δ7.71-7.63(m,2H),6.86(d,J＝9.0Hz,1H),3.14-3.06(m,2H),2.97(s,6H),2.31(t,J＝7.3Hz,2H),1.75-1.65(m,2H),1.55-1.28(m,15H)。

Dissolving the compound 3.3(514mg, 1.32mmol) in aminoethanol, adding a catalytic amount of Raney nickel, introducing 3atm of hydrogen, reacting at room temperature for 6 hours, filtering to obtain Raney nickel, and removing the solvent from the filtrate under reduced pressure to obtain the product 3.4(446mg) with the yield of 86%. UPLC-MS (ESI) theoretical value of C₂₁H₃₆N₄O₃[M+H]⁺393.28, found 393.69.

C014 was prepared according to the synthetic route in example 2 using compound 3.4 and compound 2.5 as starting materials.¹HNMR(400MHz,DMSO-d₆)δ11.12(s,1H),7.98(s,1H),7.80(dd,J＝8.4,7.4Hz,1H),7.65(t,J＝8.8Hz,1H),7.48(d,J＝7.2Hz,1H),7.38(d,J＝8.6Hz,1H),7.35-7.28(m,1H),7.27-7.16(m,1H),5.19-5.15(m,0.5H),5.11(dd, J ═ 12.9,5.4Hz,1H),5.06-4.97(s,0.5H),4.75(s,2H),4.75-4.69(m,2H),3.48(d, J ═ 12.6Hz,1H),3.32(d, J ═ 10.8Hz,1H),3.12(dd, J ═ 12.8,6.7Hz,3H),3.03-2.95(m,1H),2.94-2.84(m,1H),2.83-2.68(m,10H),2.63-2.44(m,2H),2.30(d, J ═ 14.9Hz,1H),2.23-2.12(m,3H), 2.06-2.93 (m, 2.93, 1H), 1.46-1H), 1H, 12H, 1₄₅H₅₇N₁₁O₈[M+H]⁺880.44, found 880.93.

Example 4

[C011] Preparation of N- (6- (4- (dimethylamino) -3- (((8-isopropyl-2- ((1-methyl-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) N-hexyl) -2- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) acetamide

Compound 4.1 was prepared from 2-fluoro-5-bromoxynil as the starting material, according to the synthesis of compound 3.1.¹H NMR(400MHz,CDCl₃)δ7.56(d,J＝2.5Hz,1H),7.44(dd,J＝9.1,2.5Hz,1H),6.74(d,J＝9.1Hz,1H),3.05(s,6H)。

Compound 4.2 was prepared by synthesizing compound 1.3 using compound 4.1 and compound 1.2 as starting materials.¹HNMR(400MHz,CDCl₃)δ7.84(dd,J＝5.5,3.0Hz,2H),7.71(dd,J＝5.5,3.0Hz,2H),7.49(d,J＝2.0Hz,1H),7.37(dd,J＝8.8,2.1Hz,1H),6.75(d,J＝8.8Hz,1H),3.74(t,J＝7.1Hz,2H),3.07(s,6H),2.44(t,J＝7.0Hz,2H),1.90-1.81(m,2H),1.69–1.57(m,3H)。

Compound 4.3 was prepared by synthesis of compound 1.10 starting from compound 4.2.¹H NMR(400MHz,CDCl₃)δ7.50(d,J＝2.1Hz,1H),7.36(dd,J＝8.8,2.1Hz,1H),6.74(d,J＝8.8Hz,1H),3.06(s,6H),2.73(t,J＝6.7Hz,2H),2.38(t,J＝6.7Hz,2H),1.66-1.52(m,4H)。

Compound 4.3(354mg, 1.47mmol) was dissolved in 10mL of dichloromethane, and triethylamine (305. mu.L, 2.21mmol, 1.5equiv) and (Boc) were added at 0 ℃ respectively₂O (370. mu.L, 1.61mmol, 1.1equiv), reacted at room temperature for 12 hours, the solvent was removed under reduced pressure, and the product was obtained by column chromatography (437mg), yield 87%.¹H NMR(400MHz,CDCl₃)δ7.51(d,J＝2.1Hz,1H),7.37(dd,J＝8.8,2.1Hz,1H),6.75(d,J＝8.8Hz,1H),3.20-3.13(m,2H),3.08(s,6H),2.40(t,J＝6.5Hz,2H),1.65-1.57(m,4H),1.44(s,9H)。

C011 was prepared by following the synthetic route in example 3 using compound 4.4 as a starting material.¹H NMR(400MHz,DMSO-d₆) δ 11.12(s,1H),7.96(d, J ═ 1.3Hz,1H),7.83-7.69(m,1H),7.46(d, J ═ 7.3Hz,1H),7.38(d, J ═ 8.6Hz,1H),7.34-7.08(m,3H),5.22-5.16(m,0.5H),5.14-4.98(m,1.5H),4.83(s,2H),4.77(s,2H),3.41(d, J ═ 11.3Hz,1H),3.27(d, J ═ 14.1Hz,1H),3.19-2.81(m,12H),2.7-2.64(m,3H),2.61-2.45(m,2H), 2.29-2.92 (m,2H), 2.42-2H), 1.42(m,2H), 1H), 1.06 (m, 2.06-2H), 1H, 31, 2.32(m,2H), 1H, 5H, 1H₄₄H₅₆N₁₀O₇[M+H]⁺837.43, found 837.87.

[C010] Preparation of N- (6- (3- (dimethylamino) -4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) N-hexyl) -2- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) acetamide

Referring to the synthetic route in example 4, C010 was prepared by substituting 2-fluoro-5-bromoxynil for 2-fluoro-4-bromoxynil.¹H NMR(400MHz,DMSO-d₆) δ 11.13(s,1H),7.98(d, J ═ 1.8Hz,1H),7.79(dd, J ═ 8.4,7.4Hz,1H),7.49(d, J ═ 7.2Hz,1H),7.37(d, J ═ 8.5Hz,1H),7.30-7.10(m,2H),7.05-6.90(m,1H),5.20-5.15(s,0.5H),5.11(dd, J ═ 12.9,5.4Hz,1H),5.07-4.99(m,0.5H),4.76(s,2H),4.71(t, J ═ 6.3Hz,2H),3.49(d, J ═ 11.6, 1H),3.33(d, 3.3, 3.19 (d, 3.7, 2H), 3.19 (d, 3.3.3, 2H), 3.19 (d, 3.19, 2H), 3.3.19 (d, 3, 2H), 3.19 (d, 3.7.7, 3, 2H), 3.7.7.7, 3, 10H, 1H, 7.7 (d, 1H), 3.7.7.7, 10H, 1H), 2H) 1.78(d, J ═ 13.5Hz,1H),1.56-1.48(m,2H),1.46-1.38(m,2H),1.32-1.19(m,10H), UPLC-ms (esi) theoretical values C₄₄H₅₆N₁₀O₇[M+H]⁺837.43, found 837.84.

[C015] Preparation of N- (3- (dimethylamino ((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) -7- (2- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) acetamido) heptanamide

Referring to the synthetic route in example 3, C015 was prepared by replacing 2-fluoro-5-nitrobenzonitrile with 2-fluoro-4-nitrobenzonitrile.¹H NMR(400MHz,DMSO-d₆) δ 11.13(s,1H),7.97(s,1H), 7.83-7.77(m,1H), 7.58(d, J ═ 10.2Hz,1H),7.49(d, J ═ 7.3Hz,1H),7.38(d, J ═ 8.5Hz,1H),7.23(t, J ═ 8.5Hz,1H),7.13(d, J ═ 8.5Hz,1H),5.20-5.15(m,0.5H),5.12(dd, J ═ 12.9,5.4Hz,1H),5.06-4.96(m,0.5H),4.76(s,2H),4.71-4.65(m,2H),3.49(d, J ═ 12.0, 1H),3.32 (m, 3.32H), 3.94 (d, 3.3.83-2H), 3.42H, 3.94 (d, 3.3.3.3.3H), 3.3.3.3H, 3.94 (d, 3.3H), 1H) 2.07-1.93(m,2H),1.82-1.70(m,1H),1.60-1.51(m,2H),1.48-1.38(m,2H),1.34-1.21(m,10H) UPLC-MS (ESI) with C as theoretical value₄₅H₅₇N₁₁O₈[M+H]⁺880.44, found 880.97.

Example 5

[C009] Preparation of 4- ((6- (3- (dimethylamino) -4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) n-hexyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione

Referring to the synthetic route in example 4, compound 5.1 was prepared by substituting 2-fluoro-5-bromoxynil for 2-fluoro-4-bromoxynil.

Dissolving compound 5.1(80mg, 0.126mmol) and compound 5.2(42mg, 0.151mmol, 1.2equiv) in 5mL DMF, adding DIPEA (104 μ L, 0.63mmol, 5equiv), reacting at 90 deg.C for 24 hr, adding water to the reaction solution, extracting with ethyl acetate, combining ethyl acetate layers, washing with saturated NaCl, and adding anhydrous Na₂SO₄Drying, column chromatography, and further purification by HPLC to obtain C009.¹H NMR(400MHz,DMSO-d₆) δ 11.11(s,1H),7.98(d, J ═ 1.6Hz,1H),7.61-7.44(m,1H),7.31-7.12(m,2H),7.07(d, J ═ 8.7Hz,1H),7.01(d, J ═ 7.0Hz,1H),6.98-6.91(m,1H),5.20-5.16(m,0.5H),5.08-4.98(m,1.5H),4.71(t, J ═ 6.2Hz,2H),3.49(d, J ═ 11.2Hz,1H),3.38-3.23(m,3H),3.21-3.03(m,2H),2.98(td, J ═ 9.4,6.5,2.5, 2H), 2.94(m, 1.84H), 1.31, 1H, 1.06 (m, 1.4H), 1.6, 1H), 1.5H, 1.84 (m,1H), 1.1.1H, 1H), 1H, 1H, 2H) 1.41-1.31(m,2H),1.30-1.18(m,10H). UPLC-MS (ESI) has a theoretical value of C₄₂H₅₄N₁₀O₅[M+H]⁺779.43, found 779.81.

[C016] Preparation of N- (3- (dimethylamino) -4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) -7- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) amino) heptanamide

Referring to the synthetic routes in example 3 and example 5, C016 was prepared.¹H NMR(400MHz,DMSO-d₆)δ11.11(s,1H),7.97(s,1H),7.61-7.50(m,2H),7.25-7.18(m,1H),7.14-7.06(m,2H),7.01(d,J＝7.0Hz,1H),5.19-5.15(m,0.5H),5.08-4.96(m,1.5H),4.68(t,J＝5.9Hz,2H),3.49(d,J＝12.5Hz,1H),3.36-3.24(m,3H),3.20-3.05(m,2H),2.99(dtd,J＝9.7,6.7,2.6Hz,1H),2.92-2.76(m,4H),2.76-2.65(m,6H),2.62–2.45(m,2H),2.35-2.24(m,3H),2.18(d,J＝15.1Hz,1H),2.06-1.93(m,2H),1.84-1.70(m,1H),1.62-1.53(m,2H),1.35(s,2H),1.30-1.20(m,10H)。

Example 6

[C022] Preparation of 7- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) heptanamide

Compound 6.1 was prepared by synthesis of compound 1.12 starting from compound 2.8 and 7-hydroxyheptanoic acid.¹HNMR(400MHz,CDCl₃)δ7.71(s,1H),7.52(d,J＝8.2Hz,2H) 7.21(d, J ═ 8.2Hz,2H),5.17-5.08(m,1H),4.70(d, J ═ 5.3Hz,2H),3.59(t, J ═ 6.4Hz,2H),3.07(dt, J ═ 13.8,6.9Hz,1H),3.00-2.92(m,2H),2.70-2.56(m,2H),2.47(s,3H),2.35(t, J ═ 7.3Hz,2H),2.30-2.20(m,2H),2.08-2.96(m,2H),1.76-1.64(m,2H),1.56-1.48(m,2H),1.40-1.32(m,4H),1.27(d, J ═ 6.6H), theoretical values of (lc, 5.6-5 Hz,6H), (1 lc-6H), (1H), (1.6H), (1H₂₈H₄₁N₇O₃[M+H]⁺524.33, found 524.94.

Compound 6.1(40mg, 0.0764mmol), Compound 6.2(32mg, 0.115mmol, 1.5equiv) (see Lohbeck J., Miller A. K. practical synthesis of a phenyl ligand and an enabled PROTAC reduction. bioorganic for the preparation of Compound 6.2&medicinal chemistriylethers.2016; 5260 (5260) -.¹H NMR(400MHz,DMSO-d₆) δ 11.11(s,1H),7.96(d, J ═ 0.9Hz,1H),7.79(dd, J ═ 8.4,7.4Hz,1H),7.55 to 7.48(m,3H),7.43(d, J ═ 7.2Hz,1H),7.25(d, J ═ 8.5Hz,2H),5.25 to 5.21(m,0.5H),5.12 to 4.98(m,1.5H),4.59(dd, J ═ 10.1,6.5Hz,2H),4.23 to 4.17(m,2H),3.49(d, J ═ 12.7Hz,1H),3.33(d, J ═ 11.4Hz,1H),3.21 to 3.08(m,2H), 2.97.97 (d, J ═ 12.7Hz,1H), 1H, 3.33(d, J ═ 11.4Hz,1H),3.21 to 3.08(m,2H),2.97(d, J ═ 2H, 1.7, 1.6.6.6.7H, 1H), 1H, 2H, 1H, 2H, 1H, 2H) 1.52-1.43 (m,2H),1.41-1.32(m,2H),1.26(d, J ═ 6.9Hz,6H) UPLC-ms (esi) theoretical value C₄₁H₄₉N₉O₇[M+H]⁺780.38, found 780.84.

[C017] Preparation of 7- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) amino) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) heptanamide

Referring to the synthetic routes in example 2 and example 5, C017 was prepared.¹H NMR(400MHz,DMSO-d₆)δ11.08(s,1H),7.95(s,1H),7.60-7.46(m,3H),7.25(d,J＝7.9Hz,2H),7.07(d,J＝8.4Hz,1H),7.00(d,J ═ 6.8Hz,1H),5.36-5.27(m,1H),5.26-5.17(m,0.5H),5.11-4.98(m,0.5H),4.60(s,2H),3.53-3.42(m,1H),3.36-3.23(m,3H),3.22-3.08(m,2H),3.02-2.94(m,1H),2.92-2.74(m,4H),2.62-2.42(m,2H),2.33-2.23(m,3H),2.18(d, J ═ 13.9Hz,1H),2.08-1.94(m,2H),1.85-1.71(m,1H),1.65-1.49(m,4H), 1.40-1.13H (m,10H), theoretical lc (lc-1C), (esi, 1H), and (m,0.5H)₄₁H₄₉N₉O₇[M+H]⁺779.39, found 779.32.

Example 7

[C005] Preparation of 4- ((6- (3- (dimethylamino) -4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) n-hexyl) oxy) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione

Compound 7.3 was prepared according to the synthetic route in example 4, starting from compound 7.1 (see example 4). UPLC-MS (ESI) theoretical value of C₁₅H₂₆N₂O[M+H]⁺251.20, found 251.75.

Compound 7.3(190mg, 0.759mmol) was dissolved in 10mL of anhydrous dichloromethane, TBSCl (137mg, 0.911mmol, 1.2equiv) and imidazole (77mg, 1.139mmol, 1.5equiv) were added to each solution at 0 deg.C, reacted at room temperature for 12 hours, dichloromethane was added to the reaction, washed with water several times, and anhydrous Na₂SO₄Drying, filtering, pressurizing to remove the solvent, and performing column chromatography to obtain a product 7.4(197mmol), wherein the yield is 71%. UPLC-MS (ESI) theoretical value of C₂₁H₄₀N₂OSi[M+H]⁺365.29, found 365.94.

Compound 7.6 was prepared using compound 7.4 as the starting material according to the synthetic route described in example 3.

Compound 7.6(15mg, 0.0235mmo) was dissolved in 1mL tetrahydrofuran, added with TBAF (1M solution in THF, 71. mu.L, 0.0705mmol, 3equiv), reacted at room temperature for 12 hours, then the solvent was removed under reduced pressure, and column chromatography was performed to give product 7.7(12mg) with 98% yield.¹H NMR(400MHz,CDCl₃)δ7.69(s,1H),7.20(d, J ═ 7.7Hz,1H),6.99(s,1H),6.86(d, J ═ 6.7Hz,1H),5.12-5.04(m,1H),4.83(d, J ═ 5.4Hz,2H),3.62(t, J ═ 6.6Hz,2H),3.07(dt, J ═ 13.8,6.9Hz,1H),2.91-2.82(m,2H),2.73(s,6H),2.61-2.53(m,2H),2.42-2.33(m,5H),2.22-2.13(m,2H),2.04-1.91(m,2H),1.65-1.51(m,4H),1.42-1.32(m, 1H), 1.29.7 (m,6H), theoretical lc, 0 (C), (il, 1H), 1H₂₉H₄₅N₇O₂[M+H]⁺524.36, found 524.67.

C005 was prepared by synthesizing compound C022 from compound 7.7 and compound 6.2 as starting materials.¹H NMR(400MHz,CDCl₃) δ 7.82(s,1H),7.67(t, J ═ 7.8Hz,1H),7.46(d, J ═ 7.1Hz,1H),7.41(d, J ═ 7.7Hz,1H),7.20(d, J ═ 10.7Hz,3H),5.53-5.37(m,1H),5.01-4.88(m,3H),4.16(t, J ═ 5.7Hz,2H),3.46-3.22(m,3H),3.17(s,6H),3.05(dt, J ═ 13.7,7.0Hz,1H),2.93-2.80(m,4H),2.76(d, J ═ 15.6Hz,1H),2.67(t, J ═ 7.0, 2H), 2.47-2.31.31H, 2.31H, 1H, 2.73(m, 1H), 2.31, 2.4H, 2.31, 8H) UPLC-MS (ESI) theoretical value of C₄₂H₅₃N₉O₆[M+H]⁺780.41, found 780.77.

Example 8

[C003] Preparation of 2- (2, 6-dioxopiperidin-3-yl) -4- ((7- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) n-heptyl) oxy) isoindol-1, 3-dione

P-bromobenzylamine (2g, 10.75mmol) was dissolved in 20mL of dichloromethane, triethylamine (2.24mL, 16.13mmol, 1.5equiv) was added, benzyl chloroformate was slowly added dropwise at 0 ℃ to react at room temperature for 12 hours, dichloromethane and saturated NaHCO were added to the reaction₃Washing with anhydrous Na₂SO₄Drying and column chromatography gave 8.1(3.1g) with 90% yield.¹H NMR(400MHz,CDCl₃)δ7.44(d,J＝8.3Hz,2H),7.39-7.27(m,5H),7.15(d,J＝8.0Hz,2H),5.13(s,2H),4.32(d,J＝6.0Hz,2H)。

Compound 8.4 was prepared using compound 8.1 as the starting material according to the synthetic route described in example 7.

Compound 8.4(206mg, 0.614mmol), compound 8.5(164mg, 0.675mmol, 1.1equiv) (see EP2634190a1) were dissolved in 10mL of acetonitrile, DIPEA (304 μ L, 1.842mmol, 3equiv) was added thereto, reaction was carried out at 60 ℃ for 16 hours, the solvent was removed under reduced pressure, and column chromatography was carried out to give product 8.6(305mg) in 92% yield.

Compound 8.6(305mg, 0.56mmol) was dissolved in 10mL of dichloromethane, m-CPBA (291mg, 1.69mmol, 3equiv) was added in portions, and after 4 hours at room temperature, the reaction was diluted with dichloromethane, washed several times with 0.5N aqueous NaOH solution and dried over Na₂SO₄Drying gave 8.7(270mg) in 90% yield.¹H NMR(400MHz,CDCl₃)δ7.93(s,1H),7.30(d,J＝8.0Hz,2H),7.18(d,J＝8.0Hz,2H),4.87(d,J＝5.8Hz,2H),3.59(t,J＝6.6Hz,2H),3.36(s,3H),3.25(dt,J＝13.8,6.9Hz,1H),2.63-2.56(m,2H),1.63-1.55(m,2H),1.54-1.45(m,2H),1.37-1.27(m,12H),0.89(s,9H),0.04(s,6H)。

C003 was prepared by following the synthetic route in example 7 using compound 8.7 as a starting material.¹H NMR(400MHz,DMSO-d₆) δ 11.11(s,1H),7.95(d, J ═ 1.1Hz,1H),7.82-7.75(m,1H),7.49(d, J ═ 8.6Hz,1H),7.43(d, J ═ 7.2Hz,1H),7.24(d, J ═ 7.0Hz,2H),7.13(d, J ═ 7.9Hz,2H),5.25-5.20(m,0.5H),5.11-5.01(m,1.5H),4.62(dd, J ═ 11.2,6.3Hz,2H),4.18(t, J ═ 6.2Hz,2H),3.49(d, J ═ 11.9Hz,1H),3.33(d, J ═ 11.4, 3.23H, 3.2Hz, 2H),3.49(d, J ═ 11.9, 1H), 3.7.7H, 1H, 7.7.7, 17H, 1H, 7.7 (d, 8H, 2H, 1H, 2H, 7.7, 7.7.7H, 7H, 7.9H, 7H, 7.7, 7H, 2H, 7H, 2H, 7, 2H, 7H, 2, 3H) theoretical values of 1.59-1.48(m,2H),1.48-1.38(m,2H),1.37-1.21(m,10H) UPLC-MS (ESI) are C₄₁H₅₀N₈O₆[M+H]⁺751.39, found 751.68.

[C004] Preparation of 2- (2, 6-dioxopiperidin-3-yl) -4- (2- (2- (3- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) propoxy) ethoxy) isoindole-1, 3-dione

Reference is made to example 8Synthesizing route, and preparing C004.¹H NMR(400MHz,DMSO-d₆) δ 11.11(s,1H),7.96(d, J ═ 1.4Hz,1H),7.78(t, J ═ 7.7Hz,1H),7.52(d, J ═ 8.6Hz,1H),7.44(d, J ═ 7.2Hz,1H),7.23(d, J ═ 6.2Hz,2H),7.12(d, J ═ 8.0Hz,2H),5.24-5.19(s,0.5H),5.11-4.99(m,1.5H),4.61(dd, J ═ 11.3,6.3Hz,2H),4.36-4.31(m,2H),3.82-3.78(m,2H),3.66-3.61(m,2H),3.52-3.45(m, 3.45H), 3.7 (m, 3.7H), 3.7-4.7H), 3.7 (d, 2H), 3.7H, 3.3.3.7 (d, 3.3H), 3.7 (d, 3.3H), 3.7H), 3.3.7H, 3.7H, 3.3H), 1H) 2.04-1.94(m,2H),1.82-1.67(m,3H),1.26(d, J ═ 6.9Hz,6H), UPLC-ms (esi) theoretical value C₄₁H₅₀N₈O₈[M+H]⁺783.38, found 784.12.

[C006] Preparation of 4- ((6- (3- (dimethylamino) -2- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) n-hexyl) oxy) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione

Referring to the synthetic route in example 7, C006 was prepared by substituting 2-fluoro-4-bromoxynil for 2-fluoro-6-bromoxynil.¹H NMR(400MHz,DMSO-d₆) δ 11.10(s,1H),7.94(d, J ═ 3.5Hz,1H),7.80(t, J ═ 7.9Hz,1H),7.46(dd, J ═ 10.9,7.9Hz,2H),7.40-7.28(m,2H),7.23-7.08(m,1H),5.29-5.24(m,0.5H),5.14-5.02(m,1.5H),4.82(dd, J ═ 11.2,4.6Hz,2H),4.12(t, J ═ 6.2Hz,2H),3.52(d, J ═ 11.8Hz,1H),3.39(d, J ═ 11.6Hz,1H),3.16(dt, J ═ 22.3,10.5, 3.8H, 1H), 3.65 (d, J ═ 11.6Hz,1H), 1H, 13.6H, 13.8H, 13.6H, 13(d, 2H), 3.6H, 1H, 13.6H, 2H, 13.6H, 1H, 13H, 2H, 13H, and 13H, 18H), 1.69-1.59(m,2H),1.54-1.44(m,2H), 1.41-1.13 (m,10H), UPLC-MS (ESI) theoretical value is C₄₂H₅₃N₉O₆[M+H]⁺780.41, found 780.58.

[C007] Preparation of 4- ((7- (3- (dimethylamino) -4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) n-heptyl) oxy) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione

Prepared according to the synthetic route described in example 7, replacing 5-hexyn-1-ol by 6-heptyn-1-olC007。¹HNMR(400MHz,DMSO-d₆) δ 11.11(s,1H),7.97(s,1H),7.85-7.73(m,1H),7.50(d, J ═ 8.6Hz,1H),7.44(d, J ═ 7.2Hz,1H),7.19-7.05(m,2H),6.97-6.81(m,1H),5.20-5.15(s,0.5H),5.11-4.97(m,1.5H),4.71(t, J ═ 5.9, 2H),4.18(t, J ═ 6.3Hz,2H),3.48(d, J ═ 12.4Hz,1H),3.31(d, J ═ 11.8Hz,1H),3.20-3.03(m,2H),2.98(d, 10.4Hz, 1H), 1H, 3.63, 10.15H, 1H), 3.93 (d, 1.15H, 1H),3.15, 3.3.3.3.3.3, 3, 3.3, 3, 15, 1H, 15, 1H, 1H, 3, 1.60-1.50(m,2H),1.48-1.40(m,2H),1.39-1.18(m,10H), UPLC-MS (ESI) theoretical value is C₄₃H₅₅N₉O₆[M+H]⁺794.43, found 794.55.

[C008] Preparation of 4- ((7- (2- (dimethylamino) -3- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) n-heptyl) oxy) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione

Referring to the synthetic route in example 7, C008 was prepared by substituting 2-fluoro-4-bromoxynil for 2-fluoro-3-bromoxynil and 5-hexyn-1-ol for 6-heptyn-1-ol.¹H NMR(400MHz,CDCl₃) δ 7.86(s,1H),7.70-7.64(m,1H),7.44(d, J ═ 7.2Hz,1H),7.25-7.10(m,4H),5.55-5.49(m,1H),4.92(dd, J ═ 11.9,5.0Hz,1H),4.83(s,2H),4.19(t, J ═ 6.0Hz,2H),3.52-3.23(m,3H),3.09(dd, J ═ 13.5,6.7Hz,1H),2.99(s,6H),2.92-2.73(m,5H), 2.73-2.62 (m,2H),2.51-2.34(m,2H),2.34-2.21(m,2H),2.16-1.98(m,1H), 1.70-1.82 (m,1H), 1.35 (m,1H),1.8 (1H), 1.8 (m, 8-1H), 1H, 8 (lc, 8 (1H), 1H, 8 (dl-1H), 1H, 8 (dl, 1H),1₄₃H₅₅N₉O₆[M+H]⁺794.43, found 794.52.

[C030] Preparation of 2- (2- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) ethoxy) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) acetamide

Referring to the synthetic route in example 6, C030 was prepared by substituting 7-hydroxyheptanoic acid for 2- (2- (2-hydroxyethoxy) acetic acid.¹H NMR(400MHz,CDCl₃)δ7.87(s,1H),7.78-7.72(m,1H),7.52(d,J＝7.2Hz,1H),7.36(dJ ═ 8.2Hz,2H),7.31(d, J ═ 8.4Hz,1H),7.22(d, J ═ 8.3Hz,2H),5.45-5.39(m,1H),4.85-4.66(m,3H),4.46-4.40(m,2H),4.23(s,2H),4.11-4.02(m,2H),3.43(d, J ═ 10.9Hz,2H),3.27-3.13(m,2H),3.07(dt, J ═ 13.8,6.9Hz,1H),2.86-2.77(m,4H),2.74-2.63(m,2H),2.44-2.27(m,2H),2.24-2.11(m,2H), 2.97 (m,1H), 29.97 (m, 6.6H), theoretical value of jc (lc-1H), and (d, 6H)₃₈H₄₃N₉O₈[M+H]⁺754.33, found 754.99.

[C031] Preparation of 2- (2- (2- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) ethoxy) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) acetamide

Referring to the synthetic route in example 6, C031 was prepared by replacing 7-hydroxyheptanoic acid with 2- (2- (2-hydroxyethoxy) ethoxy) acetic acid.¹H NMR(400MHz,CDCl₃) δ 7.93(s,1H),7.74-7.68(m,1H),7.55(d, J ═ 8.1Hz,2H),7.51(d, J ═ 7.3Hz,1H),7.27(d, J ═ 5.5Hz,1H),7.12(d, J ═ 7.9Hz,2H),5.48-5.42(m,1H),4.88-4.80(m,1H),4.72-4.56(m,2H),4.46-4.38(m,2H),4.09-3.98(m,4H),3.89-3.75(m,4H),3.51-3.38(m,2H),3.33-3.19(m,2H),3.13-3.04(m,1H),2.91-2.74(m,5H), 2.6-4H, 7.6 (m,6H), 1H, 6(m,6H), 1H, 6H, 15(m, 6H).₄₀H₄₇N₉O₉[M+H]⁺798.34, found 798.99.

[C032] Preparation of 2- (2- (2- (2- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) ethoxy) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) acetamide

Referring to the synthetic route in example 6, C032 was prepared by substituting 7-hydroxyheptanoic acid for 2- (2- (2-hydroxyethoxy) ethoxy) acetic acid.¹H NMR(400MHz,CDCl₃)δ7.86(s,1H),7.70-7.64(m,1H),7.58(d,J＝8.5Hz,2H),7.46(d,J＝7.2Hz,1H),7.26-7.19(m,3H),5.42–5.37(m,1H),4.89-4.81(m,1H),4.68(dd,J＝11.9,6.0Hz,2H),4.31(t,J＝4.4Hz,2H),4.13-4.08(m,2H),3.97-3.91(m,2H),3.85(t, J ═ 4.3Hz,2H),3.79-3.69(m,6H),3.43(d, J ═ 10.4Hz,2H),3.26-3.14(m,2H),3.07(dt, J ═ 13.8,7.0Hz,1H),2.86-2.66(m,6H),2.44-2.33(m,2H),2.33-2.25(m,2H),2.11-1.98(m,1H),1.29(d, J ═ 6.9Hz,6H), UPLC-ms (esi) with a theoretical value of C₄₂H₅₁N₉O₁₀[M+H]⁺842.38, found 842.94.

Example 9

[C041] Preparation of 7- (2- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindol-4-yl) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) heptanamide

6-heptynoic acid (250mg, 1.98mmol) was dissolved in 6mL DMF, cesium carbonate (322mg, 0.99mmol, 0.5equiv) was added and reacted for 20 minutes, then benzyl bromide (259. mu.L, 2.18mmol, 1.1equiv) dissolved in 1mL DMF was added dropwise and reacted at room temperature for 12 hours, ethyl acetate was added to the reaction mixture, washing with water was carried out several times, washing with saturated NaCl, anhydrous Na₂SO₄Drying and column chromatography gave 9.1(400mg) in 94% yield.¹H NMR(400MHz,CDCl₃)δ7.40-7.30(m,6H),5.12(s,2H),2.39(t,J＝7.5Hz,2H),2.21(td,J＝7.0,2.7Hz,2H),1.95(t,J＝2.7Hz,1H),1.82-1.73(m,2H),1.61-1.52(m,2H)。

Compound 9.2 was prepared by synthesis of compound 1.3 using compound 9.1 as the starting material.¹H NMR(400MHz,CDCl₃)δ7.80(d,J＝7.0Hz,1H),7.55(dd,J＝7.6,0.8Hz,1H),7.42(t,J＝7.6Hz,1H),7.38-7.29(m,5H),5.24(dd,J＝13.4,5.1Hz,1H),5.10(s,2H),4.50(d,J＝16.8Hz,1H),4.36(d,J＝16.8Hz,1H),2.94-2.76(m,2H),2.50-2.39(m,5H),2.22-2.14(m,1H),1.88-1.79(m,2H),1.69-1.60(m,2H)。

Compound 9.3 was prepared using compound 9.2 as the starting material, with reference to compound 1.4.

Compound 9.3 and compound 2.8 were used as starting materials, and C041 was prepared by synthesis of compound 1.12.¹H NMR(400MHz,DMSO-d₆)δ11.00(s,1H),7.96(d,J＝0.7Hz,1H),7.59-7.48(m,3H),7.46-7.40(m,2H),7.25(d, J ═ 8.5Hz,2H),5.25-5.21(m,0.5H),5.13(dd, J ═ 13.3,5.1Hz,1H),5.08-4.99(m,0.5H),4.59(dd, J ═ 10.0,6.4Hz,2H),4.45(d, J ═ 17.2Hz,1H),4.29(d, J ═ 17.2Hz,1H),3.48(d, J ═ 11.9Hz,1H),3.33(d, J ═ 11.4Hz,1H),3.21-3.07(m,2H),3.02-2.86(m,2H),2.81(dd, 12.40 (m,2H),7.25(d, J ═ 8.5H), 5.7.7.7.1H, 1H), 5.23.7.1H, 1H, 3.65 (d, 1H, 7.65(d, 1H, 4H) 1.39-1.29(m,4H),1.26(d, J ═ 6.9Hz,6H), UPLC-ms (esi) theoretical values C₄₁H₅₁N₉O₅[M+H]⁺750.40, found 751.08.

Example 10

[C027] Preparation of 9- ((2- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindol-4-yl) oxy) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) nonanamide

Azelaic acid (2g, 10.63mmol) was dissolved in 20mL of anhydrous tetrahydrofuran, DBU (1.59mL, 10.63mmol, 1equiv) was added, benzyl bromide (1.26mL, 10.63mmol, 1equiv) was added dropwise at 0 ℃ and reacted at room temperature for 12 hours, the solvent was removed under reduced pressure, water was added thereto, ethyl acetate was extracted, the ethyl acetate layer was combined, washed with saturated NaCl, and anhydrous Na₂SO₄Drying and column chromatography gave compound 10.1(1.45g) in 49% yield.¹H NMR(400MHz,CDCl₃)δ7.40-7.29(m,5H),5.12(s,2H),2.38-2.31(m,4H),1.69-1.56(m,4H),1.38-1.28(m,6H)。

Compound 10.1(1.45g, 5.2mmol) was dissolved in 15mL of anhydrous tetrahydrofuran, borane (1M solution in THF, 5.2mL, 5.2mmol, 1equiv) was added dropwise at 0 ℃, reaction was carried out at room temperature for 12 hours, methanol was added dropwise to the reaction to quench the reaction, the solvent was removed under reduced pressure, and column chromatography was carried out to give compound 10.2(1.35g) with a yield of 98%.

Compound 10.4 was prepared with a yield of 98% by reference to the synthesis of compound C022 starting from compound 10.2 and compound 10.3.¹H NMR(400MHz,CDCl₃)δ7.42-737(m,2H),7.37-7.30(m,5H),7.00-6.97(m,1H),5.11(s,2H),4.91(dd, J ═ 8.8,6.2Hz,1H),4.43(d, J ═ 17.6Hz,1H),4.36(d, J ═ 17.5Hz,1H),4.03(td, J ═ 6.3,1.3Hz,2H),3.63(s,3H),2.50-2.29(m,5H),2.25-2.14(m,1H),1.83-1.74(m,2H),1.70-1.61(m,2H),1.49-1.40(m,2H),1.40-1.30(m,6H), UPLC theoretical value (lc-lc) is (lc-1, 6H)₃₀H₃₈N₂O₇[M+H]⁺539.27, found 539.80.

Compound 10.4(220mg, 0.408mmol) was dissolved in 3mL of tetrahydrofuran, and TBAF (1M solution in THF, 816. mu.L, 0.816mmol, 2equiv) was added thereto, followed by reaction at room temperature for 4 hours, removal of the solvent under reduced pressure, and column chromatography gave compound 10.5(200mg) in 97% yield.¹H NMR(400MHz,DMSO-d₆)δ10.96(s,1H),7.47(t,J＝7.8Hz,1H),7.39-7.32(m,5H),7.30(d,J＝7.4Hz,1H),7.23(d,J＝8.1Hz,1H),5.13–5.06(m,3H),4.36(d,J＝17.5Hz,1H),4.21(d,J＝17.3Hz,1H),4.10(t,J＝6.4Hz,2H),2.96-2.84(m,1H),2.62-2.54(m,1H),2.49-2.38(m,1H),2.34(t,J＝7.4Hz,2H),2.02-1.93(m,1H),1.77-1.67(m,2H),1.58-1.49(m,2H),1.45-1.36(m,2H),1.33-1.21(m,6H)。

Compound 10.6 was prepared by synthesis of compound 9.3 using compound 10.5 as the starting material. UPLC-MS (ESI) theoretical value of C₂₂H₂₈N₂O₆[M+H]⁺417.19, found 417.54.

Compound C027 was prepared starting from compound 10.6 and compound 2.8, according to the synthetic route for compound 1.12.¹H NMR(400MHz,DMSO-d₆) δ 10.97(s,1H),7.96(d, J ═ 0.9Hz,1H),7.52(d, J ═ 8.5Hz,1H),7.46(t, J ═ 7.8Hz,1H),7.29(d, J ═ 7.3Hz,1H),7.25(d, J ═ 8.5Hz,2H),7.22(d, J ═ 8.1Hz,1H),5.25-5.20(m,0.5H),5.10(dd, J ═ 13.4,5.1Hz,1H),5.07-4.99(m,0.5H),4.59(dd, J ═ 9.9,6.3Hz,2H),4.36(d, J ═ 17.4Hz,1H),4.21(d, J ═ 4.2H), 4.81 (d, 17.2H), 4.2H, 3H, 2H),4.36(d, J ═ 17.4.4, 1H, 4.81, 3H, 3, 2.31-2.22(m,3H),2.18(d, J ═ 13.9Hz,1H),2.06-1.94(m,2H),1.82-1.67(m,3H),1.61-1.52(m,2H),1.47-1.37(m,2H),1.27(t, J ═ 10.8Hz,12H) theory values for C, ullc-ms (esi)₄₃H₅₅N₉O₆[M+H]⁺794.43, found 794.37.

Example 11

[C026] Preparation of 9- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) nonanamide

C026 was prepared according to the above synthetic route, referring to the experimental procedure in example 10.¹H NMR(400MHz,DMSO-d₆) δ 11.12(s,1H),7.96(d, J ═ 0.7Hz,1H),7.85-7.71(m,1H),7.54-7.48(m,3H),7.43(d, J ═ 7.2Hz,1H),7.25(d, J ═ 8.5Hz,2H),5.26-5.21(m,0.5H),5.11-4.98(m,1.5H),4.59(dd, J ═ 10.0,6.4, 2H),4.18(t, J ═ 6.3Hz,2H),3.48(d, J ═ 12.4Hz,1H),3.33(d, J ═ 10.9Hz,1H),3.23-3.08(m,2H),2.97 (J ═ 8, 13.4 Hz,1H), 3.3.3.33 (d, J ═ 10.9Hz,1H),3.23-3.08(m,2H),2.97(dt, 8, 6.4H), 3.3.3.3.3.3.3H), 3.3.3.3.3.3 (m, 3.3, 3.3.3, 3, 3.3.3.3.3, 3, 3.3, 3, 2H) theoretical values of 1.50-1.39(m,2H),1.38-1.21(m,12H) UPLC-MS (ESI) are C₄₃H₅₃N₉O₇[M+H]⁺808.41, found 808.93.

[C018] Preparation of 5- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) pentanamide

Referring to the synthetic route in example 11, azelaic acid was replaced with glutaric acid and C018 was prepared.¹H NMR(400MHz,DMSO-d₆)δ11.11(s,1H),7.96(d,J＝0.8Hz,1H),7.83-7.77(m,1H),7.56-7.49(m,3H),7.44(d,J＝7.2Hz,1H),7.26(d,J＝8.5Hz,2H),5.25-5.20(m,0.5H),5.11-4.98(m,1.5H),4.59(dd,J＝9.8,6.5Hz,2H),4.26-4.20(m,2H),3.49(d,J＝12.1Hz,1H),3.33(d,J＝10.9Hz,1H),3.21-3.07(m,2H),2.97(dt,J＝13.8,6.8Hz,1H),2.92-2.76(m,4H),2.61-2.46(m,2H),2.38(t,J＝6.3Hz,2H),2.27(d,J＝11.6Hz,1H),2.18(d,J＝15.1Hz,1H),2.06-1.96(m,2H),1.84-1.72(m,J＝10.4Hz,5H),1.26(d, J ═ 6.9Hz,6H) UPLC-ms (esi) theoretical value C₃₉H₄₅N₉O₇[M+H]⁺752.34, found 752.98.

[C019] Preparation of 5- ((2- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindol-4-yl) oxy) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) pentanamide

C019 was prepared by substituting azelaic acid for glutaric acid according to the synthetic route in example 10.¹H NMR(400MHz,DMSO-d₆) δ 10.97(s,1H),7.96(d, J ═ 0.7Hz,1H),7.52(d, J ═ 8.5Hz,2H),7.46(t, J ═ 7.8Hz,1H),7.30(d, J ═ 7.5Hz,1H),7.28-7.20(m,3H),5.25-5.20(m,0.5H),5.10(dd, J ═ 13.3,5.1Hz,1H),5.07-4.99(m,0.5H),4.60(dd, J ═ 9.9,6.4Hz,2H),4.36(d, J ═ 17.4Hz,1H),4.21(d, J ═ 17.4Hz,1H),4.14(t, J ═ 1.5, 2H),4.23 (d, J ═ 2H),3.8 (d, J ═ 1.4H, 1H), 3.4H, 3.81 (d, 3.3.3, 3.3H), 3.3.3H, 3H, 1H, 6.9, 13H, 3H, 1H, 13H, 1H, and 1H), 2.41-2.32(m,2H),2.27(d, J ═ 13.7Hz,1H),2.18(d, J ═ 15.5Hz,1H),2.06-1.92(m,2H),1.82-1.69(m,5H),1.26(d, J ═ 6.9Hz,6H), UPLC-ms (esi) with theoretical values of C₃₉H₄₇N₉O₆[M+H]⁺738.37, found 738.98.

[C020] Preparation of 6- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) hexanamide

C020 was prepared by substituting azelaic acid for adipic acid according to the synthetic route in example 11.¹H NMR(400MHz,DMSO-d₆)δ11.12(s,1H),7.96(d,J＝0.8Hz,1H),7.84-7.75(m,1H),7.55-7.48(m,3H),7.43(d,J＝7.2Hz,1H),7.25(d,J＝8.6Hz,2H),5.25-5.20(m,1H),5.11-4.97(m,1.5H),4.59(dd,J＝10.0,6.5Hz,2H),4.20(t,J＝6.3Hz,2H),3.49(d,J＝12.4Hz,1H),3.33(d,J＝11.0Hz,1H),3.25-3.07(m,2H),2.97(dt,J＝13.8,7.0Hz,1H),2.92-2.75(m,4H),2.61-2.42(m,2H),2.34-2.23(m,3H),2.18(d,J＝15.2Hz,1H),2.06-1.95(m,2H),1.82-1.72(m,3H),1.70-1.60(m,2H),1.53-1.42(m,2H),1.26(d,J＝6.9Hz,6H).UPLC-MS (ESI) theoretical value of C₄₀H₄₇N₉O₇[M+H]⁺766.36, found 766.95.

[C021] Preparation of 6- ((2- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindol-4-yl) oxy) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) hexanamide

Referring to the synthetic route in example 10, azelaic acid was replaced by adipic acid to prepare C021.¹H NMR(400MHz,DMSO-d₆) δ 10.99(s,1H),7.96(d, J ═ 0.7Hz,1H),7.52(d, J ═ 8.5Hz,2H),7.46(t, J ═ 7.8Hz,1H),7.30(d, J ═ 7.4Hz,1H),7.27-7.20(m,3H),5.26-5.20(s,0.5H),5.10(dd, J ═ 13.3,5.0Hz,1H),5.07-4.99(m,0.5H),4.59(dd, J ═ 10.1,6.3Hz,2H),4.36(d, J ═ 17.4Hz,1H),4.20(d, J ═ 17.4Hz,1H),4.11(t, J ═ 6.3H, 2H),4.36(d, J ═ 17.4Hz,1H),4.11(t, J ═ 6.3, 2H), 3.7.7.7, 7.7H, 7.7.7H, 7H, 7.7 (m,3H), 7.7.7.4H, 7.7H, 7H, 7.7H, 7H, 2.35-2.23(m,3H),2.18(d, J ═ 13.9Hz,1H),2.07-1.91(m,2H),1.82-1.70(m,3H),1.69-1.59(m,2H),1.51-1.40(m,2H),1.26(d, J ═ 6.9Hz,6H), and lc-ms (esi) theoretical values of C₄₀H₄₉N₉O₆[M+H]⁺752.38, found 752.53.

[C023] Preparation of 7- ((2- (2, 6-dioxopiperidin-3-yl) -1-oxoindol-4-yl) oxy) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) heptanamide

Referring to the synthetic route in example 10, azelaic acid was replaced by pimelic acid to prepare C023.¹H NMR(400MHz,DMSO-d₆)δ10.99(s,1H),7.96(s,1H),7.51(d,J＝8.4Hz,2H),7.45(t,J＝7.8Hz,1H),7.29(d,J＝7.4Hz,1H),7.27-7.19(m,3H),5.25-5.19(m,0.5H),5.10(dd,J＝13.3,5.0Hz,1H),5.06-4.98(m,0.5H),4.59(dd,J＝10.3,6.5Hz,2H),4.36(d,J＝17.4Hz,1H),4.20(d,J＝17.4Hz,1H),4.10(t,J＝6.2Hz,2H),3.51-3.45(m,1H),3.33(d,J＝10.5Hz,1H),3.23-3.07(m,2H),3.03-2.84(m,2H),2.81(dd,J＝13.4,4.7Hz,3H),2.62-2.38(m,2H),2.34-2.23(m,3H),2.18(d,J＝15.1Hz,1H),2.06-1.92(m,2H),1.82-1.68(m,3H),1.64-1.55(m,2H),1.50-1.40(m2H),1.40-1.31(m,2H),1.26(d, J ═ 6.9Hz,6H), UPLC-ms (esi) theoretical values C₄₁H₅₁N₉O₆[M+H]⁺766.40, found 766.91.

[C024] Preparation of 8- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) octanamide

C024 was prepared by substituting azelaic acid for suberic acid according to the synthetic route in example 11.¹H NMR(400MHz,DMSO-d₆) δ 11.11(s,1H),7.96(s,1H), 7.82-7.75(m,1H), 7.54-7.47(m,3H),7.43(d, J ═ 7.2Hz,1H),7.25(d, J ═ 8.5Hz,2H),5.26-5.20(m,0.5H),5.11-4.99(m,1.5H),4.59(dd, J ═ 10.1,6.3Hz,2H),4.19(t, J ═ 6.3Hz,2H),3.48(d, J ═ 11.8Hz,1H),3.33(d, J ═ 11.3Hz,1H),3.22-3.08(m,2H),3.02-2.93(m,1H),2.93 (m, 2.93-2H), 4.62 (m, 18H), 1H, 18-1H), 1H (m,1H), 1H, 1, 18, 1H, 18, 1H, 18, 1H, 18, 4H, 3H, 18, 1H, 2H) 1.41-1.29(m,4H),1.26(d, J ═ 6.9Hz,6H), UPLC-ms (esi) theoretical values C₄₂H₅₁N₉O₇[M+H]⁺794.39, found 795.04.

[C025] Preparation of 8- ((2- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindol-4-yl) oxy) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) octanamide

C025 was prepared by substituting azelaic acid for suberic acid according to the synthetic route in example 10.¹H NMR(400MHz,DMSO-d₆)δ10.99(s,1H),7.96(d,J＝0.9Hz,1H),7.52(d,J＝8.4Hz,2H),7.46(t,J＝7.8Hz,1H),7.29(d,J＝7.4Hz,1H),7.27-7.19(m,3H),5.25-5.18(s,0.5H),5.10(dd,J＝13.3,5.1Hz,1H),5.07-4.98(m,0.5H),4.59(dd,J＝10.2,6.5Hz,2H),4.36(d,J＝17.4Hz,1H),4.20(d,J＝17.4Hz,1H),4.09(t,J＝6.3Hz,2H),3.48(d,J＝12.1Hz,1H),3.33(d,J＝11.6Hz,1H),3.22-3.07(m,2H),3.02-2.92(m,1H),2.95-2.76(m,4H),2.59-2.41(m,2H),2.31-2.23(m,3H),2.18(d,J＝14.8Hz,1H),2.05-1.93(m,2H),1.81-1.68(m,3H),1.62–1.52(m,2H),1.47-1.38(m,2H),1.38-1.28(m,4H),1.25(d,J＝6.9Hz6H) UPLC-MS (ESI) theoretical value of C₄₂H₅₃N₉O₆[M+H]⁺780.41, found 781.07.

[C028] Preparation of 10- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) decanamide

C028 was prepared by replacing azelaic acid with sebacic acid according to the synthetic route of example 11.¹H NMR(400MHz,DMSO-d₆) δ 11.11(s,1H),7.95(s,1H),7.83-7.75(m,1H),7.54-7.47(m,3H),7.43(d, J ═ 7.2Hz,1H),7.25(d, J ═ 8.5Hz,2H),5.26-5.20(m,0.5H),5.11-4.99(m,1.5H), 4.63-4.55 (m,2H),4.18(t, J ═ 6.1Hz,2H),3.48(d, J ═ 12.4Hz,1H),3.33(d, J ═ 11.7Hz,1H),3.22-3.08(m,2H), 3.01-2.93(m,1H), 2.92-2.76(m,4H), 2.63-2H, 44(m,2H), 3.49-3.3.3.3H), 3.01-2.93(m,1H), 2.06 (m,1H), 3.51, 3.6H), 3.6H, 1H), 3.49-3.6H, 1H, 3.6H, 3.6, 3, 1.36-1.22(m,14H). UPLC-MS (ESI) theoretical value of C₄₄H₅₅N₉O₇[M+H]⁺822.43, found 822.90.

[C029] Preparation of 10- ((2- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindol-4-yl) oxy) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) decanamide

By referring to the synthetic route in example 10, azelaic acid was replaced by sebacic acid to prepare C029.¹H NMR(400MHz,DMSO-d₆) δ 10.97(s,1H),7.95(s,1H),7.52(d, J ═ 8.4Hz,2H),7.46(t, J ═ 7.7Hz,1H),7.29(d, J ═ 7.5Hz,1H),7.25(d, J ═ 8.5Hz,2H),7.22(d, J ═ 8.2Hz,1H),5.26-5.20(m,0.5H),5.14-4.98(m,1.5H),4.64-4.53(m,2H),4.36(d, J ═ 17.5Hz,1H),4.21(d, J ═ 17.4Hz,1H),4.09(t, J ═ 6.4Hz,2H),3.49(d, J ═ 13.2, 1H), 3.95 (s,1H), 3.23.23H, 3.23 (d, 3.6, 2H), 3.23H, 3.23(m, 3.6, 2H), 3.23H, 3.6, 3.5H, 3, 3.6, 3, 2H, 3, 2H, 1, 2.07-1.92(m,2H),1.85-1.67(m,3H),1.61-1.50(m,2H),1.46-1.37(m,2H),1.37-1.19(m,14H) UPLC-MS (ESI) with theoretical value of C₄₄H₅₇N₉O₆[M+H]⁺808.44, found 808.78.

[C033] Preparation of 8- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) -N- (3- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) octanamide

Referring to the synthetic routes in example 2 and example 6, C033 was prepared.¹H NMR(400MHz,DMSO-d₆) δ 11.11(s,1H),7.97(d, J ═ 0.9Hz,1H),7.84-7.77(m,1H),7.56(s,1H),7.52-7.45(m,2H),7.44(d, J ═ 7.2Hz,1H),7.23(t, J ═ 7.8Hz,1H),7.02-6.97(m,1H),5.24-5.19(m,0.5H),5.07(dd, J ═ 12.8,5.3Hz,1H),5.04-4.97(m,0.5H),4.62(dd, J ═ 10.1,6.2Hz,2H),4.19(t, J ═ 6.4Hz,2H),3.47(d, J ═ 13.0, 3.0, J ═ 10.1,6.2Hz,2H),4.19(t, J ═ 6.4Hz,2H),3.47(d, J ═ 13.8, 13.2H, 3.2H), 2H, 13.9, 2H, 13.2H, 2H, 13.9-2H, 2H, 2, 1H) 2.05-1.94(m,2H),1.81-1.69(m,3H),1.62-1.53(m,2H),1.45(s,2H),1.39-1.29(m,4H),1.26(d, J ═ 6.9Hz,6H), UPLC-ms (esi) with the theoretical value C₄₂H₅₁N₉O₇[M+H]⁺794.39, found 794.96.

[C039] Preparation of 8- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) -N- (4- (((8-isopropyl-2- (((R) -piperidin-3-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) octanamide

Referring to the synthetic route in example 11, C039 was prepared.¹H NMR(400MHz,DMSO-d₆) δ 11.09(s,1H),7.97(s,1H), 7.82-7.76 (m,1H), 7.58-7.48 (m,3H),7.43(d, J ═ 7.2Hz,1H),7.26(d, J ═ 8.4Hz,2H), 5.26-5.20(m, 1H),5.07(dd, J ═ 12.8,5.3Hz,1H),4.60(d, J ═ 6.0Hz,2H),4.19(t, J ═ 6.3Hz,2H), 3.40-3.27 (m,2H), 3.11-2.94 (m,3H), 2.92-2.81 (m,1H), 2.62-2.43(m, 2H),2.28(t, J ═ 7.3, 2H), 1.63-1H, 1.80 (m,1H), 1.50-1H), 1.80 (m,1H), 1H, 1₄₀H₄₉N₉O₇[M+H]⁺780.38, found 780.99.

[C040] Preparation of 8- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) -N- (4- (((8-isopropyl-2- (((S) -piperidin-3-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) octanamide

Referring to the synthetic route in example 11, C040 was prepared.¹H NMR(400MHz,DMSO-d₆) δ 11.11(s,1H),7.97(s,1H),7.79(dd, J ═ 8.4,7.4Hz,1H), 7.56-7.47 (m,3H),7.43(d, J ═ 7.2Hz,1H),7.26(d, J ═ 8.5Hz,2H), 5.27-5.20 (m,1H),5.07(dd, J ═ 12.8,5.4Hz,1H),4.60(d, J ═ 6.2Hz,2H),4.19(t, J ═ 6.4Hz,2H), 3.40-3.26 (m,2H), 3.12-2.94 (m,3H), 2.92-2.80 (m,1H), 2.62-2.44(m,2H), 28.28 (t, J ═ 2H),1.7 (m,1H), 1.6H, 1H, 26(m ═ 6H), 1H, 6H, 1H, 6H, 1H, 6H, 1H₄₀H₄₉N₉O₇[M+H]⁺780.38, found 781.14.

Example 12

[C045] Preparation of N1- ((R) -1- ((2S,4R) -4-hydroxy-2- ((4- (4-methylthiazol-5-yl) benzyl) carboxamido) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxybutan-2-yl) -N8- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) octanediamide

Compound 12.1 (see Buckley D.L., Van Mulle I., Gareiss P.C., et al. targeting the von Hippel-Lindau E3ubiquitin ligand using small molecule deletion the VHL/HIF-1alpha interaction. journal of American chemical society.2012; 134(10): 4465-.¹H NMR(400MHz,CDCl₃) δ 8.67(s,1H),7.41-7.29(m,4H),6.21(d, J ═ 8.8Hz,1H),4.68(dd, J ═ 10.4,5.7Hz,1H),4.59-4.44(m,4H),4.33(dd, J ═ 15.0,5.3Hz,1H),4.07(d, J ═ 11.4Hz,1H),3.66-3.57(m,4H),2.53-2.42(m,4H),2.28(t, J ═ 7.5Hz,2H),2.20-2.08(m,3H),1.64-1.52(m,4H),1.34-1.22(m,4H),0.93(s,9H), esi (lc-esi) theoretical value is 8.8Hz,1H, 0.93(s,9H)₃₁H₄₄N₄O₆S[M+H]⁺601.30, found 601.78.

Compound 12.2(40mg, 0.0666mmol) was dissolved in 2mL of methanol, 2mL of 1M aqueous lithium hydroxide was added, the mixture was reacted at room temperature for 2 hours, the pH was adjusted to 3 with 1N HCl, dichloromethane extraction was performed, the dichloro layers were combined, and anhydrous Na was added₂SO₄Drying, filtration and removal of the solvent under reduced pressure gave the crude compound 12.3(40 mg). UPLC-MS (ESI) theoretical value of C₃₀H₄₂N₄O₆S[M+H]⁺587.28, found 587.81.

C045 was prepared by synthesizing compound 1.12 using compound 12.3 and compound 2.8 as starting materials.¹H NMR(400MHz,DMSO-d₆) δ 9.00(s,1H),7.95(s,1H),7.52(d, J ═ 8.1Hz,2H),7.44-7.35(m,4H),7.25(d, J ═ 8.3Hz,2H),5.26-5.19(m,0.5H),5.09-4.98(m,0.5H),4.64-4.56(m,2H),4.53(d, J ═ 9.4Hz,1H),4.46-4.38(m,2H),4.37-4.32(m,1H),4.21(dd, J ═ 15.8,5.3Hz,1H),3.69-3.55(m,2H),3.54-3.42(m,1H),3.38-3.26(m,1H), 3.3.3.7 (m,3H), 3.7 (m, 3.7, 2H),3.7 (d, 2H),3.7 (m,2H), 3.7 (d, 2H),3.7 (d, 2H),3.7, 3.6H), 3.7, 1H), 8.7,4.5Hz,1H),1.77(d, J ═ 13.0Hz,1H),1.60-1.40(m,4H),1.34-1.20(m,10H),0.92(s,9H), UPLC-ms (esi) with theoretical values of C₅₁H₆₉N₁₁O₆S[M+H]⁺964.52, found 965.08.

Example 13

[C049] Preparation of N1- (4- (N- (3-cyano-4-methyl-1H-indol-7-yl) sulfonamide) benzyl) -N8- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) octanediamide

C049 was prepared by substituting compound 12.1 for compound 13.1 according to the synthetic route in example 12.¹HNMR(400MHz,DMSO-d₆)δ8.17(d,J＝3.2Hz,1H),7.95(s,1H),7.65(d,J＝8.4Hz,2H),7.52(d,J＝8.5Hz,2H),7.34(d,J＝8.4Hz,2H),7.25(d,J＝8.5Hz,2H),6.76(d,J＝8.3Hz,1H),6.57(d,J＝7.7Hz,1H),5.26-5.20(m,05H),5.07-4.98(m,0.5H),4.64-4.55(m,2H),4.28(d, J ═ 5.9Hz,2H),3.49(d, J ═ 12.7Hz,1H),3.33(d, J ═ 11.3Hz,1H),3.21-3.08(m,2H),2.97(dt, J ═ 13.9,6.9Hz,1H),2.81(dd, J ═ 13.1,4.7Hz,3H),2.55(s,3H),2.30-2.22(m,3H),2.21-2.09(m,3H),2.06-1.95(m,1H),1.81-1.71(m,1H),1.59-1.46(m,4H), 1.32-1.32 (m,10H), theoretical values of (lc, 1H), (esi, 1H), (1.59-1H), (10C) and (lc)₄₆H₅₅N₁₁O₅S[M+H]⁺87441, found 87497.

[C046] Preparation of N1- (4- (N- (3-cyano-4-methyl-1H-indol-7-yl) sulfonamido) benzyl) -N10- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) sebacamide

C046 was prepared by replacing suberic acid with sebacic acid according to the synthetic route in example 13.¹H NMR(400MHz,DMSO-d₆) δ 8.16(d, J ═ 3.1Hz,1H),7.95(d, J ═ 0.7Hz,1H),7.65(d, J ═ 8.4Hz,2H),7.52(d, J ═ 8.4Hz,2H),7.34(d, J ═ 8.4Hz,2H),7.26(d, J ═ 8.5Hz,2H),6.76(d, J ═ 8.4Hz,1H),6.59(d, J ═ 7.7Hz,1H), 5.25-5.20(m,0.5H), 5.07-5.00 (m,0.5H),4.60(dd, J ═ 9.5,6.4Hz,2H),4.28(d, J ═ 5.9, 2H),3.52 (m, 3.5H), 3.60 (dd, 3.5H), 3.81-2H), 3.7.7H, 3.7H, 3.2H, 3.9, 3.2H, 3.81-2H, 3.7H, 3.9, 3.5H, 3.9, 3.4H, 3.9, 3.4H, 3.9, 3, 3.9, 3, 1H) theoretical values of 1.60-1.45 (m,4H), 1.31-1.18 (m,14H) UPLC-MS (ESI) are C₄₈H₅₉N₁₁O₅S[M+H]⁺902.44, found 902.65.

[C047] Preparation of N- (4- (N- (3-cyano-4-methyl-1H-indol-7-yl) sulfonamido) benzyl) -2- (2- ((4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) amino) -2-oxoethoxy) ethoxy) acetamide

Referring to the synthetic route in example 13, suberic acid was replaced with 3, 6-dioxasuberic acid to afford C047.¹HNMR(400MHz,DMSO-d₆)δ8.16(d,J＝3.1Hz,1H),7.95(s,1H),7.64(d,J＝8.4Hz,2H),7.56(d,J＝8.5Hz,2H),7.36(d,J＝8.3Hz,2H),7.27(d,J＝8.5Hz,2H),6.75(d,J＝8.4Hz,1H),6.57(d,J＝7.7Hz,1H),5.25–5.19(m,0.5H), 5.07-4.98(m,0.5H), 4.64-4.56(m,2H), 4.34(d, J ═ 6.2Hz,2H),4.07(s,2H),4.00(s,2H), 3.71-3.62 (m,4H), 3.51-3.44 (m,1H), 3.36-3.28 (m,1H), 3.21-3.08(m,2H),2.97(dt, J ═ 13.7,6.8Hz,1H),2.80(dd, J ═ 12.0,4.7Hz,3H),2.54(s,3H), 2.30-2.23(m, 1H), 2.21-2.14 (m,1H), 2.06-1.95(m,1H), 1.82(m, 1H),1.9 (m,1H), 1H, 26(m, 9H), theoretical lc, 26 (C, 26H), and C (d, 1H)₄₄H₅₁N₁₁O₇S[M+H]⁺878.37, found 879.29.

[C048] Preparation of N1- (4- (N- (3-cyano-4-methyl-1H-indol-7-yl) sulfonylamino) benzyl) -N6- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) adipamide

C048 was prepared by replacing suberic acid with adipic acid according to the synthetic route in example 13.¹H NMR(400MHz,DMSO-d₆) δ 8.16(d, J ═ 3.1Hz,1H),7.95(s,1H),7.65(d, J ═ 8.3Hz,2H),7.53(d, J ═ 8.4Hz,2H),7.35(d, J ═ 8.3Hz,2H),7.26(d, J ═ 8.5Hz,2H),6.76(d, J ═ 8.0Hz,1H),6.58(d, J ═ 7.7Hz,1H), 5.25-5.20(m,0.5H), 5.07-4.99(m,0.5H),4.60(dd, J ═ 9.5,6.6Hz,2H),4.29(d, J ═ 5.8Hz,2H), 3.52-3.44 (m, 3.36H), 3.7.7 (m, 3.7H), 3.7-7H, 3.7H, 3H, 3.7 (m-3H), 3.7H, 3H, 3.7H, 3H, 4H) 1.25(d, J ═ 6.8Hz,6H). UPLC-ms (esi) theoretical value C₄₄H₅₁N₁₁O₅S[M+H]⁺846.38, found 846.83.

Example 14

[C034] Preparation of N- (3- (1H-imidazol-1-yl) -4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) -8- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) octanamide

According to the above synthetic route, with reference to the synthetic routes in example 2 and example 11, preparationTo obtain C034.¹H NMR(400MHz,DMSO-d₆) δ 11.09(s,1H),9.39(d, J ═ 10.3Hz,1H),9.19(dd, J ═ 12.7,6.0Hz,1H),8.04(s,1H),7.98-7.91(m,2H),7.86(s,1H),7.83-7.77(m,1H),7.64-7.54(m,2H),7.50(d, J ═ 8.5Hz,1H),7.43(d, J ═ 7.2Hz,1H),5.19-5.14(m,0.5H),5.11-4.96(m,1.5H),4.46(d, J ═ 5.5Hz,2H),4.19(t, J ═ 6.3, 2H),3.51(d, 1.92H), 3.12H, 3.31, 3.12H, 3H, 3.31H, 3.12H, 3H, 3.31H, 3H, 3.12H, 3H, 3.5H, 3H, 1H) 2.11-1.94(m,2H),1.88-1.68(m,3H),1.64-1.53(m,2H),1.50-1.41(m,2H),1.40-1.15(m,10H) UPLC-MS (ESI) with C as theoretical value₄₅H₅₃N₁₁O₇[M+H]⁺860.41, found 860.52.

[C035] Preparation of N- (2- (dimethylamino) -3- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) -8- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) octanamide

Referring to the synthetic route in example 14, C035 was prepared by substituting 2-fluoro-4-nitrobenzonitrile for 2-fluoro-3-nitrobenzonitrile and imidazole for dimethylamine hydrochloride.¹H NMR(400MHz,DMSO-d₆) δ 11.11(s,1H),7.98(s,1H),7.83-7.77(m,1H),7.63-7.55(m,1H),7.51(d, J ═ 8.7Hz,1H),7.44(d, J ═ 7.2Hz,1H),7.03(t, J ═ 8.0Hz,1H),6.95-6.89(m,1H),5.18-5.14(m,0.5H),5.08(dd, J ═ 12.9,5.5Hz,1H),5.01-4.92(m,0.5H),4.74-4.67(m,2H),4.21(t, J ═ 6.3Hz,2H),3.54-3.47(m,1H),3.34-3.27 (m), 3.3.98 (m,1H), 3.3.3 (m,2H), 2.3.3.3.3.3.3-2H (m,2H), 2m, 2H (m, 2.3.3.3, 2H), 2H, 1H) 2.05-1.91(m,2H),1.82-1.69(m,3H),1.67-1.57(m,2H),1.51-1.43(m,2H),1.42-1.33(m,4H),1.31-1.19(m,6H) UPLC-MS (ESI) with a theoretical value of C₄₄H₅₆N₁₀O₇[M+H]⁺837.43, found 837.58.

[C036] Preparation of N- (3- (dimethylamino) -4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) -8- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) octanamide

Reference toThe synthetic route in example 14, substituting imidazole for dimethylamine hydrochloride, yielded C036.¹H NMR(400MHz,DMSO-d₆) δ 11.10(s,1H),7.97(s,1H),7.83-7.77(m,1H),7.53-7.48(m,2H),7.43(d, J ═ 7.2Hz,1H),7.23-7.17(m,1H),7.12-7.05(m,1H),5.19-5.15(m,0.5H),5.07(dd, J ═ 13.1,5.5Hz,1H),5.04-4.96(m,0.5H),4.68(t, J ═ 5.8Hz,2H),4.19(t, J ═ 6.5Hz,2H),3.54-3.47(m,1H),3.35-3.27(m,1H),3.19-3.05(m,2H),3.03 (m,2H), 94.92 (m,2H), 7.7.83-7.7.7.7 (m,1H), 7.17(m,1H), 7.05(m,2H), 7.6.6.6 (m,2H), 2H), 7.6 (m,2H),1.81-1.69(m,3H),1.62-1.53(m,2H),1.51-1.41(m,2H), 1.41-1.30 (m,4H),1.27(dd, J ═ 6.9,2.9Hz,6H), UPLC-ms (esi) with theoretical value C₄₄H₅₆N₁₀O₇[M+H]⁺837.43, found 837.72.

Example 15

[C042] Preparation of 5- (3- (2- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindol-4-yl) propoxy) -N- (4- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) pentanamide

Compound 15.1(786mg, 4.5mmol) was dissolved in 10mL of methylene chloride and 10mL of 35% (w/w) aqueous sodium hydroxide solution, tetrabutylammonium chloride (1.25g, 4.5mmol, 1equiv) was added, bromopropyne (800. mu.L, 9mmol, 2equiv) was added dropwise at 0 ℃ to react at room temperature for 12 hours, methylene chloride was added to the reaction, and the reaction was washed with water several times and anhydrous Na₂SO₄Drying and column chromatography gave 15.2(342mg) in 36% yield.¹H NMR(400MHz,CDCl₃)δ4.11(d,J＝2.4Hz,2H),3.51(t,J＝6.2Hz,2H),2.40(t,J＝2.4Hz,1H),2.25–2.20(m,2H),1.68–1.57(m,4H),1.43(s,9H)。

C042 was prepared according to the synthetic route of example 9 using compound 15.2 as a starting material.¹H NMR(400MHz,DMSO-d₆)δ10.98(s,1H),7.95(s,1H),7.59–7.49(m,3H),7.44(d,J＝4.2Hz,2H),7.26(d,J＝8.4Hz,2H),5.25–5.21(m,1H),5.12(dd,J＝13.3,5.0Hz,0.5H),5.07–5.00(m,0.5H) 4.60(dd, J ═ 9.4,6.5Hz,2H),4.45(d, J ═ 17.1Hz,1H),4.30(d, J ═ 17.1Hz,1H), 3.53-3.45 (m,1H), 3.42-3.30 (m,5H), 3.22-3.08(m,2H), 3.02-2.85 (m,2H),2.81(dd, J ═ 12.3,4.5Hz,3H),2.68(t, J ═ 7.6Hz,2H), 2.63-2.54 (m,1H), 2.47-2.36 (m,1H), 2.34-2.23(m,3H), 2.22-2.14 (m,1H), 2.08-1.96 (m,2H),1.88 (m,1H),1.56 (m,6H), 1H, 26-1H), 1H (m, 26H), 1H, 26(m,1H), 1H, 26(m, 26, 1H), and (lc, 1H), theoretical values (C, 1H) are as shown in the following₄₂H₅₃N₉O₆[M+H]⁺780.41, found 780.66.

Example 16

[C037] Preparation of 8- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) -N- (4- (((2- (((R) -1-hydroxybutyl-2-yl) amino) -8-isopropylpyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) octanamide

Compound 2.6(110mg, 0.239mmol), compound 16.1 (90. mu.L, 0.955mmol, 4equiv) were dissolved in 3mL NMP, reacted at 120 ℃ for 12 hours, added with ethyl acetate, washed several times with anhydrous Na₂SO₄Drying and column chromatography gave 16.2(105mg) in 94% yield. UPLC-MS (ESI) theoretical value of C₂₄H₃₅N₇O₃[M+H]⁺470.28, found 470.96.

C037 was prepared by following the synthetic route in example 11 using compound 27.2 as the starting material.¹H NMR(400MHz,DMSO-d₆) δ 11.09(s,1H),7.98(s,1H), 7.83-7.75(m,1H), 7.56-7.47 (m,3H),7.43(d, J ═ 7.2Hz,1H),7.32(d, J ═ 7.5Hz,2H),5.07(dd, J ═ 12.9,5.4Hz,1H), 4.67-4.51 (m,2H),4.19(t, J ═ 6.3Hz,2H), 4.01-3.90 (m,1H), 3.55-3.40 (m,2H), 3.00-2.80 (m,2H), 2.62-2.45 (m,2H),2.28(t, J ═ 7.3Hz,2H), 2.05-1.97 (m,1H),1.82 (m,1H), 1.52(m, 1H), 3.52 (m,1H), 3H, 5-3.52 (m,2H),3.52 (m,3H), 3H, 1H, 5-3H), 3H, 1H, 5(m,3H), 3H, 5-2H), 3H, 5(m,3H), 3H, 5H, 3H), 3H), 3H, 5₄₀H₄₉N₉O₇[M+H]⁺768.38, found 769.04.

[C038] Preparation of 8- ((2- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindol-4-yl) oxy) -N- (4- (((2- (((S) -1-hydroxybutyl-2-yl) amino) -8-isopropylpyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) octanamide

Referring to the synthetic route in example 16, compound 27.1 was replaced with S- (+) -2-amino-1-butanol to prepare C038.¹H NMR(400MHz,DMSO-d₆) δ 11.09(s,1H),7.99(s,1H), 7.83-7.75(m,1H), 7.57-7.47 (m,3H),7.43(d, J ═ 7.2Hz,1H),7.32(d, J ═ 7.3Hz,2H),5.07(dd, J ═ 12.9,5.3Hz,1H), 4.69-4.51 (m,2H),4.19(t, J ═ 6.3Hz,2H), 4.01-3.92 (m,1H), 3.55-3.41 (m,2H), 3.00-2.81 (m,2H), 2.61-2.45(m,2H), 2.28(t, J ═ 7.3Hz,2H), 2.05-1.97 (m,1H),1.80 (m, 1.70H), 1.42-2.45 (m,2H),2.28(t, J ═ 7.3Hz,2H), 2.05-1.97 (m,1H),1.80 (m,1H), 3H, 1.52(m, 3H), 3H, 1H, 3H), 3H, 1H, 3H, 5(₄₀H₄₉N₉O₇[M+H]⁺768.38, found 768.63.

[C043] Preparation of N1- (2- (dimethylamino) -3- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) -N8- ((S) -1- ((2S,4R) -4-hydroxy-2- ((4- (4-methylthiazol-5-yl) benzyl) carboxamido) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxybutan-2-yl) octanediamide

Referring to the synthetic route in example 12, C043 was prepared. UPLC-MS (ESI) theoretical value of C₅₃H₇₄N₁₂O₆S[M+H]⁺1007.56, found 1007.80.

[C044] Preparation of (2S,4R) -1- ((S) -2- (7- (2- (dimethylamino) -3- (((8-isopropyl-2- ((1-methylpiperidin-4-yl) oxy) pyrazolo [1,5-a ] [1,3,5] triazin-4-yl) amino) methyl) phenyl) heptanamide) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide

Referring to the synthetic route in example 4, C044 was prepared. UPLC-MS (ESI) theoretical value of C₅₂H₇₃N₁₁O₅S[M+H]⁺964.55, found 964.71.

By some of the methods described above, the following compounds may also be synthesized:

example 17: proliferative growth inhibitory Activity of Compounds on triple negative Breast cancer cells

1. Experimental methods

Human triple-negative breast cancer cells MDA-MB-468 and BT549 used in the present invention were purchased from Shanghai cell bank using L-15 medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% double antibody, and at 37 deg.C, without CO₂The culture chamber of (1) was used to culture MDA-MB-468 cells, and the DMEM/HIGH GLUCOSE medium supplemented with 20% FBS and 1% double antibody was used to culture BT549 at 37 deg.C and 5% CO₂Is grown in the environment of (a).

In the cell activity test experiment, 90 μ L of cell suspension with appropriate concentration is added to a 96-well cell culture plate according to the growth of cells, each compound to be tested is diluted with a corresponding culture medium in a gradient manner, and 10 μ L of the diluted compound is takenAdded to 90. mu.L of cells, then 5% CO at 37 ℃₂In the case of the medium-cultured cells, the cell proliferation was analyzed by WST-8, WST-8 was reduced to a yellow formazan product by lactate dehydrogenase in the cells, 10. mu.L of WST-8 reagent (DOJINDO) was added to the cells, the reaction was carried out at 37 ℃ for 1 hour or more, DMSO-treated cells were used as a positive control, OD450nm was read by a microplate reader, and the data was processed by GraphPad Prism6 software. NT: this is not indicated as a test. Activity results: expression of cellular Activity IC₅₀>20 μ M, expression of cell Activity 1 μ M<IC₅₀<20 μ M, expression of cell Activity IC₅₀<1μM。

2. Results of the experiment

Based on the test method for the cell growth inhibition activity of the compound, the compound has good inhibition activity on cell growth in triple negative breast cancer cell strains MDA-MB-468 and BT 549. Therefore, the compound of the invention can be used as a novel degradation agent of related targets in triple negative breast cancer cells.

Example 18: proliferation and growth inhibitory activity of compounds on hematological tumor cells and other solid tumor cells

The experimental method is the same as the cell testing method, and the related cell activity testing results are shown in the following table:

from the above table, it can be found that the compounds of the present invention (C024, C025 and C026) have high activity in blood tumor cell lines, such as diffuse large B-cell lymphoma cell line (OCI-LY-10), multiple myeloma cell line (MM.1S) and human acute lymphoblastic leukemia T-lymphocyte cell line (CCRF-CEM), and IC₅₀<100 nM. The compounds (C024, C025) of the invention have good activity in human prostate cancer cell line (PC-3) and human colon cancer cell line (HCT116), respectively, and can be used as a novel CDK-targeting degradation agent for preventing and treating diseases related to abnormal activity of CDK, such as solid tumor and hematological tumor.

Example 19: CDK degradation by compound detected by Western Blot

1. Experimental methods

In the Western blot experiment, cells were cultured in 6-well plates, the corresponding cells were treated with different drug concentrations, the cells were collected and lysed with RIPA lysate, 5-10 μ g of lysate was run on SDS-PAGE, then proteins were transferred to PVDF membrane, the protein of interest was detected with the corresponding antibody, actin or Tubulin was used as an internal reference, and the following is the information of the antibody used in this paper: CDK1 (abin, abs132093), CDK2 (abin, abs100510), CDK4(CST,12790), CDK5 (abin, abs100524), CDK9(CST,2316), Actin (abin, abs132001), and Tubulin (abin, abs131993 a).

2. Results of the experiment

As can be seen from the results in fig. 1, the compounds of the invention are mainly very selective and have a very good degradation effect on CDK9, whereas no significant degradation effect on other proteins of the CDK family is found. Therefore, the compounds of the present invention may be used as a novel degradant targeting CDK9 in triple negative breast cancer cells.

Claims

1. A compound represented by the general formula (I), a tautomer, an enantiomer, a diastereomer, a racemate, a metabolic precursor, a pharmaceutically acceptable salt, an ester, a prodrug, or a hydrate thereof:

wherein,

selected from the following groups:

the G is₁、G₂、G₃Each independently optionally from-C (R)_b1) or-N ═ or; wherein R is_b1Selected from hydrogen, halogen and C_1-4A hydrocarbyl group;

z is-CH₂-or-C (═ O) -;

R_a1is hydrogen, fluoro or methyl;

R_a2is hydrogen or methyl;

R_c1is hydrogen or fluorine;

Or X is absent;

m1 is 0, 1, 2, 3,4, 5,6, 7, 8,9 or 10;

m2 is 0, 1, 2, 3,4, 5 or 6;

m3 is 0, 1, 2, 3,4 or 5;

m4 is 0, 1, 2, 3,4 or 5;

m5 is 0, 1, 2, 3,4, 5 or 6;

m6 is 0, 1, 2, 3 or 4;

m7 is 0, 1, 2, 3,4, 5,6 or 7;

is selected from C_6-12Aryl, 5-8 membered cycloalkyl, 5-8 membered heterocyclyl or heteroaryl containing 1-3 heteroatoms selected from N, O and S, 5-8 membered cycloalkylC_6-12Aryl, 5-8 membered heterocyclyl or heteroarylo C containing 1-3 heteroatoms selected from N, O and S_6-12An aryl group;

R₄selected from hydrogen atoms, halogens, C_1-6Alkoxy, cyano, amino, nitro, C_1-6Alkyl radical, C_3-10CycloalkanesA group, a 5-to 8-membered heterocyclic group, C_6-10Aryl, 5-6 membered heteroaryl, -NHC (O) R₅、-NHC(O)OR₆、-NR₇R₈Wherein said alkyl or alkoxy is optionally further substituted by one or more groups selected from halogen, hydroxy, C₁-C₆Alkoxy, cyano, nitro substituent;

n is 0, 1, 2, 3, 4;

R₁is hydrogen, deuterium or CH₃；

e is selected from-O-, -CO-NH-, -NH-CO-, -O-CH₂–、–CO–NH–CH₂–、–NR₆–、–NR₆–CH₂–、–CH₂–NR₆-, -S-, -CH (OH) -or-CH₂-; preferably E is selected from-O-or-NH-;

2. The compound according to claim 1, wherein the compound of formula (I) is selected from the group consisting of the following formulae, and tautomers, enantiomers, diastereomers, racemates, metabolic precursors, pharmaceutically acceptable salts, esters, prodrugs or hydrates thereof:

wherein R is₁、R₂、R₃、E、R₄、n、X, L, Y and

is as defined in claim 1.

3. The compound according to claim 1, wherein the compound of formula (I) is selected from the group consisting of the following formulae, and tautomers, enantiomers, diastereomers, racemates, metabolic precursors, pharmaceutically acceptable salts, esters, prodrugs or hydrates thereof:

wherein R is₃-E is selected from the group consisting of:

R₁is hydrogenDeuterium or CH₃；

R₂Is hydrogen or substituted or unsubstituted C₁-C₃Alkyl, wherein said substitution is substituted with one or more substituents selected from the group consisting of: halogen, cyano, nitro, hydroxy and amino;

n is 0, 1, 2;

x, L, Y and

is as defined in claim 1.

4. The compound according to claim 1, wherein the compound of formula (I) is selected from the group consisting of the following formulae, and tautomers, enantiomers, diastereomers, racemates, metabolic precursors, pharmaceutically acceptable salts, esters, prodrugs or hydrates thereof:

R₃-E is selected from the group consisting of:

R₁is hydrogen, deuterium or CH₃；

R₂Is hydrogen or substituted or unsubstituted C_1-3Alkyl, wherein said substitution is substituted with one or more substituents selected from the group consisting of: halogen, cyano, nitro, hydroxy and amino;

n is 0, 1, 2;

selected from the following groups:

Wherein W1 and W2 are independently and preferably selected from-O-, -CH₂-、

X、Y、

m1, m2, m3, m4, m5, m6, m7 are as defined in claim 1.

5. The compound according to claim 1, wherein the compound of formula (I) is selected from the group consisting of the following formulae, and tautomers, enantiomers, diastereomers, racemates, metabolic precursors, pharmaceutically acceptable salts, esters, prodrugs or hydrates thereof:

wherein R is₁、R₂、R₃E, X, L, Y and

is as defined in claim 1.

6. The compound according to claim 1, wherein the compound of formula (I) is selected from the group consisting of the following formulae, and tautomers, enantiomers, diastereomers, racemates, metabolic precursors, pharmaceutically acceptable salts, esters, prodrugs or hydrates thereof:

wherein R is₁、R₂、R₃、E、R₄、n、X、L、Y、R_c1、R_a2、R_a3And R_a4Is as defined in claim 1.

7. The compound according to claim 1, wherein the compound of formula (I) is selected from the group consisting of the following formulae, and tautomers, enantiomers, diastereomers, racemates, metabolic precursors, pharmaceutically acceptable salts, esters, prodrugs or hydrates thereof:

wherein R is₁、R₂、R₃、E、R₄、n、X, L, Y and

is as defined in claim 1.

8. A compound according to claim 1, wherein the compound of formula (I) is selected from the group consisting of the following compounds, and tautomers, enantiomers, diastereomers, racemates, metabolic precursors, pharmaceutically acceptable salts, esters, prodrugs or hydrates thereof:

9. a process for the preparation of a compound of formula (I), said process being selected from one of the following processes: the first synthesis method comprises the following steps:

wherein R is₂、R₃、E、X₁、X₂And X₇Is as defined in claim 1;

n₁is an integer of 0 to 12;

n₂is an integer of 0 to 6;

b, reacting the compound 1C with the compound 1D under the conditions of diisopropyl azodicarboxylate and triphenylphosphine to obtain 1E;

and a second synthesis method comprises the following steps:

wherein R is₂、R₃、E、X₁、X₂And X₇Is as defined in claim 1;

u is-O-or-CH₂-；

n₃Is an integer of 0 to 9, n₄Is an integer of 0 to 5;

step E, carrying out condensation reaction on the compound 2D and the compound 1A to obtain a compound 2E;

the third synthesis method comprises the following steps:

wherein R is₂、R₃、E、X₁、X₂And X₇Is as defined in claim 1;

n₅is an integer of 0 to 12, n₆Is an integer of 0 to 6;

i, reacting a compound 3C with a compound 3F under the condition of N, N-diisopropylethylamine to obtain a compound 3G;

the synthesis method comprises the following steps:

wherein R is₂、R₃、E、X₁、X₂And X₇Is as defined in claim 1;

z is methylene or carbonyl;

n₇is an integer of 0 to 12, n₈Is an integer of 0 to 6;

j, reacting the compound 4A with the compound 4B under the conditions of diisopropyl azodicarboxylate and triphenylphosphine to obtain 4C;

step m, carrying out condensation reaction on the compound 1A and the connecting chain 4F to obtain a compound 4G;

the synthesis method comprises the following steps:

wherein R is₂、R₃、E、X₁、X₂And X₇Is as defined in claim 1;

n₉is an integer of 0 to 10, n₁₀Is an integer of 0 to 6;

step p: carrying out condensation reaction on the compound 5D and the compound 1A to obtain a compound 5E;

the synthesis method comprises the following steps:

wherein R is₂、R₃、Ra3、Ra4、E、X₁、X₂And X₇Is as defined in claim 1;

n₁₁is an integer of 0 to 6, n₁₂Is an integer of 0 to 10;

step s: carrying out condensation reaction on the compound 6D and the compound 1A to obtain a compound 6E;

the synthesis method comprises the following steps:

wherein R is₂、R₃、E、X₁、X₂And X₇Is as defined in claim 1;

n₁₃is an integer of 0 to 9，n₁₄Is an integer of 0 to 5;

step w: condensing the compound 7C and the compound 1A under the conditions of 2- (7-benzotriazole oxide) -N, N, N ', N' -tetramethylurea hexafluorophosphate and N, N-diisopropylethylamine to obtain a compound 7D;

step x: the compound 2C is subjected to palladium-carbon catalysis under the condition of normal pressure hydrogen to obtain a compound 7E;

the synthesis method comprises the following steps:

wherein R is₂、R₃、E、X₁、X₂And X₇Is as defined in claim 1;

n₁₅is an integer of 0 to 9, n₁₆Is an integer of 0 to 5;

step z 2: reacting the compound 8D under the conditions of imidazole and tert-butyldimethylsilyl chloride to obtain a compound 8E;

10. A pharmaceutical composition comprising a therapeutically effective amount of one or more compounds selected from the group consisting of compounds represented by general formula (i) as claimed in any one of claims 1 to 8, and tautomers, enantiomers, diastereomers, racemates, metabolic precursors, pharmaceutically acceptable salts, esters, prodrugs or hydrates thereof, and pharmaceutically acceptable adjuvants.

11. Use of a compound of general formula (i) as defined in any one of claims 1 to 8, and tautomers, enantiomers, diastereomers, racemates, metabolic precursors, pharmaceutically acceptable salts, esters, prodrugs or hydrates thereof, or a pharmaceutical composition as defined in claim 10, as a targeted CDK degrading agent.

12. Use of a compound of general formula (i) as defined in any one of claims 1 to 8, and tautomers, enantiomers, diastereomers, racemates, metabolic precursors, pharmaceutically acceptable salts, esters, prodrugs or hydrates thereof, or a pharmaceutical composition according to claim 10 thereof, for the manufacture of a medicament for the prevention and/or treatment of a disease or condition associated with abnormal activity of CDKs.

13. Use according to claim 12, wherein the diseases associated with abnormal CDK activity include solid tumors of breast, colon, prostate, small cell lung, non-small cell lung, and hematological tumors of acute lymphocytic leukemia, B cell lymphoma, T cell lymphoma, myeloma, acute and chronic myelogenous leukemias, promyelocytic leukemia.