CDK inhibitor compounds

Abstract

The present disclosure relates to CDK4 inhibitor compounds represented by formula (I) or pharmaceutically acceptable salts thereof, a pharmaceutical composition containing same, and the use thereof in preventing or treating CDK4-mediated diseases.

Description

CDK inhibitor compounds

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of and priority to the following Chinese invention patent applications, the entire contents of which are hereby incorporated by reference in their entirety:

[0003] Patent application No. 202410319396.X filed with the State Intellectual Property Office on March 19, 2024;

[0004] Patent application No. 202411079678.3 submitted to the State Intellectual Property Office on August 7, 2024. Technical Field

[0005] The present disclosure belongs to the field of medicine and relates to a cyclin-dependent kinase (CDK) inhibitor compound or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the same, and the use of the compound as a CDK inhibitor in preventing or treating related diseases. Background Art

[0006] The development of tumors is associated with an imbalance in the expression of multiple oncogenes and tumor suppressor genes. The functional effects of nearly all oncogenes and tumor suppressor genes ultimately converge on the cell cycle. Therefore, tumors can be considered a type of cell cycle disease (CCD), and regulating or blocking the cell cycle is one approach to treating tumors. Currently, many molecules involved in cell cycle regulation have been discovered, among which cyclin-dependent kinases (CDKs) are core molecules in the cell cycle regulatory network.

[0007] CDKs are a group of serine / threonine protein kinases. CDKs drive the cell cycle by chemically acting on serine / threonine proteins, and work synergistically with cyclins, making them important factors in cell cycle regulation.

[0008] Among the CDK subtypes involved in the cell cycle, CDK4 / 6 plays an irreplaceable role. Cancer-related cell cycle mutations mainly exist in the G1 phase and G1 / S phase transition process. CDK4 / 6 binds to cyclin D (CvclinD) to form a kinase-active complex, which releases the bound transcription factor E2F through phosphorylation of the tumor suppressor gene Rb product pRb, initiates the transcription of genes related to the S phase, prompts cells to pass the checkpoint, and transfers from the G1 phase to the S phase.

[0009] However, clinical applications have also demonstrated that CDK4 / 6 inhibitors can cause adverse reactions such as gastrointestinal and / or hematological toxicity, and may lead to acquired drug resistance over time. New research reports suggest that hematological side effects from CDK4 / 6 inhibitors may be related to CDK6 inhibition, and CDK4 has been identified as a single oncogenic factor in multiple breast cancers. Therefore, selective CDK4 inhibitors may offer improved safety and efficacy compared to CDK4 / 6 inhibitors. The present invention is dedicated to developing novel selective CDK4 inhibitors. DETAILED DESCRIPTION

[0010] The present disclosure relates to a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

[0011] in:

[0012] X and Y are independently selected from N and CR 1 ;

[0013] X 1 and X 2 independently selected from N, C and CH;

[0014] Ring A is selected from 4-10 membered heterocyclic rings and 5-12 membered heteroaromatic rings, wherein the 4-10 membered heterocyclic rings and 5-12 membered heteroaromatic rings are optionally substituted by one or more R a replace;

[0015] Ring B is selected from C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-12 membered heteroaryl, the C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-12 membered heteroaryl are optionally substituted by one or more R b replace;

[0016] R 1 Selected from hydrogen, halogen, hydroxyl, C1-C 10 Alkyl, C2-C 10 Alkenyl and C2-C 10 Alkynyl, the hydroxyl, C1-C 10 Alkyl, C2-C 10 Alkenyl and C2-C 10 Alkynyl is optionally substituted with one or more R 1a replace;

[0017] R 2 Selected from hydrogen, halogen, hydroxyl, amino, mercapto, cyano, C1-C 10 Alkyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C10 Aryl and 5-10 membered heteroaryl, the hydroxyl, amino, thiol, C1-C 10 Alkyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl are optionally substituted by one or more R 2a replace;

[0018] R 3 、R 4 and R 7 independently selected from hydrogen, halogen, hydroxy, cyano, C1-C 10 Alkyl, C1-C 10 Alkoxy and C1-C4 haloalkyl;

[0019] R 5 Selected from hydrogen, C2-C 10 Alkenyl, C2-C 10 Alkynyl and C1-C 10 Alkyl, the C2-C 10 Alkenyl, C2-C 10 Alkynyl and C1-C 10 The alkyl group is optionally replaced by one or more R 5a replace;

[0020] R 6 Selected from hydrogen, halogen, hydroxy, cyano, C1-C 10 Alkyl, C2-C 10 Alkenyl and C2-C 10 Alkynyl, the hydroxyl, C1-C 10 Alkyl, C2-C 10 Alkenyl and C2-C 10 Alkynyl is optionally substituted with one or more R 6a replace;

[0021] Every R 1a independently selected from halogen, hydroxy, cyano, amino and C1-C 10 alkyl;

[0022] Every R 2a Independently selected from halogen, hydroxy, cyano, amino, C1-C8 alkyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl, the hydroxyl, amino, C1-C8 alkyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl are optionally substituted by one or more R c replace;

[0023] Every R5a 、R 6a independently selected from halogen, hydroxy, cyano and amino;

[0024] Every R a independently selected from halogen, hydroxy, cyano, amino, C1-C4 alkyl, C1-C4 haloalkyl and C1-C4 alkoxy;

[0025] Every R b independently selected from halogen, hydroxy, amino, cyano, -C(O)-R e 、-OC(O)-R f 、-S(O)2-R e 、C2-C 10 Alkenyl, C2-C 10 Alkynyl, C1-C 10 Alkyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl, the hydroxyl, amino, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C1-C 10 Alkyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl are optionally substituted by one or more R d replace;

[0026] Every R c and R d independently selected from cyano, amino, halogen, hydroxy, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl, the amino, hydroxyl, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl are optionally substituted by one or more R g replace;

[0027] Every R e 、R f and R g Independently selected from halogen, amino, C1-C4 alkyl, C1-C4 alkoxy and 4-10 membered heterocyclyl, wherein the amino, C1-C4 alkyl, C1-C4 alkoxy and 4-10 membered heterocyclyl are optionally substituted by amino, halogen, hydroxyl and C1-C4 alkyl;

[0028] One or more hydrogen atoms of the compound are optionally deuterium atoms.

[0029] In some embodiments, at least one of X and Y is CR 1 .

[0030] In some embodiments, X is N and Y is CR 1 .

[0031] In some embodiments, X and Y are both CR 1 .

[0032] In some embodiments, X 1 and X 2 At least one is C.

[0033] In some embodiments, X 1 and X 2 Both are C.

[0034] In some embodiments, X 1 It is N, X 2 It’s C.

[0035] In some embodiments, X 1 It's C, X 2 It's N.

[0036] In some embodiments, Ring A is selected from a 4-7 membered heterocyclic ring and a 5-10 membered heteroaryl ring, wherein the 4-7 membered heterocyclic ring and the 5-10 membered heteroaryl ring are optionally substituted by one or more R a replace.

[0037] In some embodiments, Ring A is selected from a 5-6 membered heterocyclic ring and a 5-6 membered heteroaryl ring, wherein the 5-6 membered heterocyclic ring and the 5-6 membered heteroaryl ring are optionally substituted by one or more R a replace.

[0038] In some embodiments, ring A is selected from pyrazole ring, imidazole ring, triazole ring, thiazole ring, oxazole ring, dihydrofuran ring and pyridine ring, and the pyrazole ring, imidazole ring, triazole ring, thiazole ring, oxazole ring, dihydrofuran ring and pyridine ring are optionally substituted by one or more R a replace.

[0039] In some embodiments, ring A is selected from a triazole ring, a thiazole ring, an oxazole ring, a dihydrofuran ring and a pyridine ring, wherein the triazole ring, the thiazole ring, the oxazole ring, the dihydrofuran ring and the pyridine ring are optionally substituted by one or more R a replace.

[0040] In some embodiments, each R a Independently selected from halogen, hydroxy, cyano, amino, C1-C4 alkyl.

[0041] In some embodiments, Selected from

[0042] In some embodiments, Selected from

[0043] In some embodiments, Selected from

[0044] In some embodiments, Ring B is selected from 4-10 membered heterocyclyl and 5-12 membered heteroaryl, wherein the 4-10 membered heterocyclyl and 5-12 membered heteroaryl are optionally substituted by one or more R b replace.

[0045] In some embodiments, Ring B is selected from 4-7 membered heterocyclyl and 5-10 membered heteroaryl, wherein the 4-7 membered heterocyclyl and 5-10 membered heteroaryl are optionally substituted by one or more R b replace.

[0046] In some embodiments, Ring B is selected from 5-6 membered heterocyclyl and 5-6 membered heteroaryl, wherein the 5-6 membered heterocyclyl and 5-6 membered heteroaryl are optionally substituted by one or more R b replace.

[0047] In some embodiments, Ring B is selected from tetrahydropyranyl, piperidinyl, and pyridinyl, wherein the tetrahydropyranyl, piperidinyl, and pyridinyl are optionally substituted with one or more R b replace.

[0048] In some embodiments, Ring B is selected from tetrahydropyranyl and pyridinyl, wherein the tetrahydropyranyl and pyridinyl are optionally substituted with one or more R b replace.

[0049] In some embodiments, each R b independently selected from halogen, hydroxy, amino, cyano, -C(O)-R e 、-OC(O)-R f 、C2-C 10 Alkenyl, C2-C 10 Alkynyl, C1-C 10 Alkyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl, the hydroxyl, amino, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C1-C 10 Alkyl, C3-C 12Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl are optionally substituted by one or more R d replace;

[0050] In some embodiments, each R b independently selected from halogen, hydroxy, -C(O)-R e 、-OC(O)-R f 、-S(O)2-R e 、C1-C 10 Alkyl and 4-10 membered heterocyclic group, the hydroxyl group, C1-C 10 The alkyl group and the 4-10 membered heterocyclic group are optionally substituted by one or more R d replace.

[0051] In some embodiments, each R b independently selected from halogen, hydroxy, -C(O)-R e 、-OC(O)-R f 、-S(O)2-R e , C1-C4 alkyl and 5-6 membered heterocyclic group, wherein the hydroxyl group, C1-C4 alkyl and 5-6 membered heterocyclic group are optionally substituted by one or more R d replace.

[0052] In some embodiments, each R b independently selected from halogen, hydroxy, -C(O)-R e 、-OC(O)-R f 、C1-C 10 Alkyl and 4-10 membered heterocyclic group, the hydroxyl group, C1-C 10 The alkyl group and the 4-10 membered heterocyclic group are optionally substituted by one or more R d replace.

[0053] In some embodiments, each R b independently selected from halogen, hydroxy, -C(O)-R e 、-OC(O)-R f , C1-C4 alkyl and 4-7 membered heterocyclic group, wherein the hydroxyl group, C1-C4 alkyl and 4-7 membered heterocyclic group are optionally substituted by one or more R d replace.

[0054] In some embodiments, each R b independently selected from fluoro, hydroxy, -C(O)-R e 、-OC(O)-R f 、-S(O)2-R e , methyl, piperidinyl and piperazinyl, wherein the hydroxyl, methyl, piperidinyl and piperazinyl groups are optionally substituted by one or more R dreplace.

[0055] In some embodiments, each R b independently selected from fluoro, hydroxy, -C(O)-R e 、-OC(O)-R f , methyl, piperidinyl and piperazinyl, wherein the hydroxyl, methyl, piperidinyl and piperazinyl groups are optionally substituted by one or more R d replace.

[0056] In some embodiments, each R e Independently selected from amino, C1-C4 alkyl, C1-C4 alkoxy and 4-7 membered heterocyclyl, said amino, C1-C4 alkyl, C1-C4 alkoxy and 4-7 membered heterocyclyl are optionally substituted by amino, halogen, hydroxyl and C1-C4 alkyl.

[0057] In some embodiments, each R e Independently selected from amino, C1-C4 alkoxy and 4-7 membered heterocyclyl, said amino, C1-C4 alkoxy and 4-7 membered heterocyclyl are optionally substituted by amino, halogen, hydroxyl, C1-C4 alkyl.

[0058] In some embodiments, each R e Independently selected from methyl, N(CH3)2, methoxy and morpholinyl.

[0059] In some embodiments, each R e Independently selected from N(CH3)2, methoxy and morpholinyl.

[0060] In some embodiments, each R f Independently selected from 4-10 membered heterocyclic groups, wherein the 4-10 membered heterocyclic groups are optionally substituted by amino, halogen, hydroxyl, or C1-C4 alkyl.

[0061] In some embodiments, each R f Independently selected from morpholinyl, the morpholinyl is optionally substituted by amino, halogen, hydroxyl, C1-C4 alkyl.

[0062] In some embodiments, each R f are independently selected from morpholinyl.

[0063] In some embodiments, each R c and R d independently selected from amino, halogen, hydroxy, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10Aryl and 5-10 membered heteroaryl, the amino, hydroxyl, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl are optionally substituted by one or more R g replace;

[0064] In some embodiments, each R d Independently selected from C1-C8 alkyl and 4-10 membered heterocyclic group, the C1-C8 alkyl and 4-10 membered heterocyclic group are optionally substituted by one or more R g replace.

[0065] In some embodiments, each R d independently selected from methyl and piperazinyl, said methyl and piperazinyl being optionally substituted by one or more R g replace.

[0066] In some embodiments, each R g Independently selected from C1-C4 alkyl and halogen, the C1-C4 alkyl is optionally substituted by amino, halogen, or hydroxy.

[0067] In some embodiments, each R g Independently selected from C1-C4 alkyl, wherein the C1-C4 alkyl is optionally substituted by amino, halogen, or hydroxy.

[0068] In some embodiments, each R g Independently selected from C1-C4 alkyl and halogen.

[0069] In some embodiments, each R g Independently selected from ethyl and fluoro.

[0070] In some embodiments, R g For ethyl.

[0071] In some embodiments, each R b Independently selected from fluorine, hydroxyl,

[0072] In some embodiments, each R b Independently selected from fluorine, hydroxyl,

[0073] In some embodiments, Ring B is selected from

[0074] In some embodiments, Ring B is

[0075] In some embodiments, R 1 is selected from hydrogen and halogen.

[0076] In some embodiments, R 1 Selected from hydrogen and fluorine.

[0077] In some embodiments, R 1 It's hydrogen.

[0078] In some embodiments, R 2 Selected from amino, C1-C 10 Alkyl, C3-C 12 Cycloalkyl and 4-10 membered heterocyclic groups, the amino, C1-C 10 Alkyl, C3-C 12 Cycloalkyl and 4-10 membered heterocyclic groups are optionally substituted by one or more R 2a replace.

[0079] In some embodiments, R 2 is selected from amino, C1-C4 alkyl, C3-C6 cycloalkyl and 4-7 membered heterocyclic groups, wherein the amino, C1-C4 alkyl, C3-C6 cycloalkyl and 4-7 membered heterocyclic groups are optionally substituted by one or more R 2a replace.

[0080] In some embodiments, R 2 Selected from C1-C 10 Alkyl and 4-10 membered heterocyclic group, the C1-C 10 The alkyl group and the 4-10 membered heterocyclic group are optionally substituted by one or more R 2a replace.

[0081] In some embodiments, R 2 Selected from C1-C4 alkyl and 4-7 membered heterocyclic group, wherein the C1-C4 alkyl and 4-7 membered heterocyclic group are optionally substituted by one or more R 2a replace.

[0082] In some embodiments, R 2 is selected from methyl, ethyl, isopropyl, cyclopentyl, amino and tetrahydropyrrolyl, wherein the methyl, ethyl, isopropyl, cyclopentyl, amino and tetrahydropyrrolyl are optionally substituted by one or more R 2a replace.

[0083] In some embodiments, R 2 is selected from methyl, ethyl, isopropyl and tetrahydropyrrolyl, wherein the methyl, ethyl, isopropyl and tetrahydropyrrolyl are optionally substituted by one or more R 2a replace.

[0084] In some embodiments, each R2a Independently selected from hydroxyl, amino, C1-C8 alkyl and 4-10 membered heterocyclic groups, wherein the hydroxyl, amino, C1-C8 alkyl and 4-10 membered heterocyclic groups are optionally substituted by one or more R c replace.

[0085] In some embodiments, each R 2a independently selected from hydroxyl, amino, C1-C4 alkyl and 4-7 membered heterocyclic groups, wherein the hydroxyl, amino, C1-C4 alkyl and 4-7 membered heterocyclic groups are optionally substituted by one or more R c replace.

[0086] In some embodiments, each R 2a Independently selected from hydroxyl, amino and 4-10 membered heterocyclic groups, wherein the hydroxyl, amino and 4-10 membered heterocyclic groups are optionally substituted by one or more R c replace.

[0087] In some embodiments, each R 2a Independently selected from hydroxyl, amino and 4-7 membered heterocyclic groups, wherein the hydroxyl, amino and 4-7 membered heterocyclic groups are optionally substituted by one or more R c replace.

[0088] In some embodiments, each R 2a independently selected from hydroxy, amino, tetrahydropyrrolyl, piperidinyl, methyl and morpholinyl, wherein the hydroxy, amino, tetrahydropyrrolyl, piperidinyl, methyl and morpholinyl are optionally substituted by one or more R c replace.

[0089] In some embodiments, each R 2a independently selected from hydroxy, amino, tetrahydropyrrolyl and morpholinyl, wherein the hydroxy, amino, tetrahydropyrrolyl and morpholinyl are optionally substituted by one or more R c replace.

[0090] In some embodiments, each R 2a Independently selected from hydroxy, amino, methyl, The hydroxyl, amino, methyl, Optional one or more R c replace.

[0091] In some embodiments, each R 2a Independently selected from hydroxyl, amino, The hydroxyl group, amino group, Optional one or more R c replace.

[0092] In some embodiments, each R 2aIndependently selected from hydroxyl, methyl and The hydroxyl group, methyl group and Optional one or more R c replace.

[0093] In some embodiments, each R c Independently selected from cyano, halogen, hydroxy, C1-C8 alkyl, wherein the C1-C8 alkyl is optionally substituted with one or more halogens.

[0094] In some embodiments, each R c Independently selected from hydroxy, C1-C8 alkyl, said C1-C8 alkyl being optionally substituted with one or more halogens.

[0095] In some embodiments, each R c Independently selected from cyano, fluoro, hydroxy, methyl and ethyl, said methyl and ethyl being optionally substituted with one or more fluoro groups.

[0096] In some embodiments, each R c Independently selected from hydroxy, methyl and ethyl, said methyl and ethyl being optionally substituted with one or more fluorine groups.

[0097] In some embodiments, R 2 Selected from methyl, aminomethyl, dimethylamino,

[0098] In some embodiments, R 2 Selected from methyl, aminomethyl,

[0099] In some embodiments, R 2 Selected from aminomethyl, In some embodiments, R 3 and R 4 independently selected from hydrogen, halogen, hydroxy, cyano, C1-C 10 Alkyl and C1-C4 haloalkyl.

[0100] In some embodiments, R 3 and R 4 are independently selected from hydrogen and halogen.

[0101] In some embodiments, R 3 and R 4 are independently selected from hydrogen, chlorine and fluorine.

[0102] In some embodiments, R 3 is selected from chlorine and fluorine, R 4 For hydrogen.

[0103] In some embodiments, R 3 is fluorine, R 4 For hydrogen.

[0104] In some embodiments, R 5 For hydrogen.

[0105] In some embodiments, R 6 Selected from hydrogen and C1-C 10 Alkyl, the C1-C 10 The alkyl group is optionally substituted with one or more R 6a replace.

[0106] In some embodiments, R 6 Selected from C1-C 10 Alkyl, the C1-C 10 The alkyl group is optionally substituted with one or more R 6a replace.

[0107] In some embodiments, R 6 Selected from C1-C4 alkyl, the C1-C4 alkyl is optionally substituted by one or more R 6a replace.

[0108] In some embodiments, R 6 is selected from hydrogen and isopropyl.

[0109] In some embodiments, R 6 It is isopropyl.

[0110] In some embodiments, R 7 For hydrogen.

[0111] In some embodiments, the compound of formula (I) of the present application or its stereoisomer or its pharmaceutically acceptable salt is selected from the compound of formula (II) or its stereoisomer or its pharmaceutically acceptable salt,

[0112] Among them, X 3 and X 4 One of them is CH and the other is N; X 1 and X 2 Independently selected from C and N; X, Y, R 2 、R 3 、R 4 、R 5 、R 6 and Ring B are as defined above.

[0113] In some embodiments, the compound of formula (I) of the present application or its stereoisomer or its pharmaceutically acceptable salt is selected from the compound of formula (III) or its stereoisomer or its pharmaceutically acceptable salt,

[0114] Among them, X 5 Selected from N and CR 8 ; R 8 is selected from hydrogen, halogen, cyano, hydroxy, amino, C1-C4 alkyl, C1-C4 haloalkyl and C1-C4 alkoxy;

[0115] R 1 、R 2 、R 3 、R 6 and Ring B are as defined above, R 6 It is preferably not H.

[0116] In some embodiments, the compound of formula (I) of the present disclosure, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from the following compounds or pharmaceutically acceptable salts thereof,

[0117] In another aspect, the present disclosure provides a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0118] In another aspect, the present disclosure provides a method for treating a disease mediated by CDK in an individual (e.g., a mammal), comprising administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof to an individual (e.g., a mammal, preferably a human) in need of such treatment.

[0119] In another aspect, the present disclosure provides use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for preventing or treating a CDK-mediated disease.

[0120] In another aspect, the present disclosure provides use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in preventing or treating a CDK-mediated disease.

[0121] In another aspect, the present disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for use in preventing or treating a CDK-mediated disease.

[0122] In some embodiments, the CDK-mediated disease is a CDK4-mediated disease.

[0123] In some embodiments, the CDK-mediated disease is a tumor.

[0124] Definitions and Explanations of Terms

[0125] Unless otherwise indicated, the terms used in this disclosure have the following meanings. The definitions of groups and terms described in this disclosure, including their definitions as examples, exemplary definitions, preferred definitions, definitions described in tables, and definitions of specific compounds in the examples, may be combined and coupled with each other in any manner. A particular term should not be considered as undefined or unclear unless specifically defined, but should be understood according to its ordinary meaning in the art. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

[0126] In this article Indicates the attachment site.

[0127] The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and diastereomers.

[0128] The diagrammatic representations of racemates or enantiomerically pure compounds herein are from Maehr, J. Chem. Ed. 1985, 62: 114-120. Unless otherwise indicated, the solid and imaginary bonds are wedge-shaped. To express the absolute configuration of a stereocenter, use direct real bonds and direct virtual bonds. Indicates the relative configuration of a stereocenter (such as the cis-trans configuration of an alicyclic compound).

[0129] The compounds of the present invention may have asymmetric atoms such as carbon atoms, sulfur atoms, nitrogen atoms, phosphorus atoms or asymmetric double bonds, so that the compounds of the present invention may exist in specific geometric or stereoisomeric forms. Specific geometric or stereoisomeric forms may be cis and trans isomers, E and Z geometric isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures or other mixtures thereof, such as mixtures enriched in enantiomers or diastereomers, all of which are within the definition of the compounds of the present invention and mixtures thereof. Additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms or asymmetric phosphorus atoms may be present in substituents such as alkyl groups, and all of which are within the definition of the compounds of the present invention and mixtures thereof. Compounds of the present disclosure containing an asymmetric atom can be isolated in optically pure or racemic forms. Optically pure forms can be resolved from racemic mixtures or synthesized by using chiral starting materials or reagents.

[0130] The term "substituted" refers to the replacement of any one or more hydrogen atoms on a particular atom by a substituent, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is an oxo (i.e., =O), it means that two hydrogen atoms are replaced. Oxo does not occur on aromatic groups.

[0131] The term "optionally" or "optionally" refers to that the event or situation described subsequently may or may not occur, and the description includes that the event or situation occurs and that the event or situation does not occur. For example, ethyl is "optionally" substituted by one or more halogens, meaning that ethyl can be unsubstituted (CH2CH3), monosubstituted (CH2CH2F, CH2CH2Cl etc.), polysubstituted (CHFCH2F, CH2CHF2, CHFCH2Cl, CH2CHCl2 etc.) or fully substituted (CF2CF3, CF2CCl3, CCl2CCl3 etc.). It will be appreciated by those skilled in the art that for any group comprising one or more substituents, any sterically impossible existence and / or incomposable replacement or substitution pattern will not be introduced.

[0132] When any variable (such as R a 、R b ) appears more than once in the composition or structure of a compound, its definition is independent in each case. For example, if a group is represented by two R b is replaced, then each R b There are independent options.

[0133] In this article, C m -C n It means having an integer number of carbon atoms in the range mn. For example, "C1-C 10 ” means that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, or 10 carbon atoms.

[0134] The term "alkyl" refers to a group of the formula C n H 2n+1 The term "C1-C 10The term "alkyl" is understood to mean a straight-chain or branched saturated hydrocarbon radical having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. Specific examples of the alkyl radical include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl alkyl or 1,2-dimethylbutyl, etc.; the term "C1-C8 alkyl" can be understood to mean an alkyl group having 1 to 8 carbon atoms; the term "C1-C6 alkyl" can be understood to mean an alkyl group having 1 to 6 carbon atoms, and specific examples include but are not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc. The term "C1-C4 alkyl" can be understood to mean a straight-chain or branched saturated alkyl group having 1 to 4 carbon atoms. The term "C1-C3 alkyl" can be understood to mean a straight-chain or branched saturated alkyl group having 1 to 3 carbon atoms. The "C1-C 10 The term "alkyl" may include "C1-C6 alkyl", "C1-C4 alkyl" or "C1-C3 alkyl", and the "C1-C6 alkyl" may further include "C1-C4 alkyl" or "C1-C3 alkyl". The term "C1-C4 haloalkyl" refers to a C1-C4 alkyl substituted by one or more halogens such as F, Cl, Br or I, including single substitution, multiple substitution or complete substitution. The term "C1-C4 hydroxyalkyl" refers to a C1-C4 alkyl substituted by one or more hydroxy groups, including single substitution, multiple substitution or complete substitution.

[0135] The term "alkoxy" refers to a group generated by the loss of a hydrogen atom from a hydroxyl group of a straight-chain or branched alcohol, and can be understood as "alkyloxy" or "alkyl-O-". The term "C1-C 10 "Alkoxy" can be understood as "C1-C 10 Alkyloxy" or "C1-C 10 The term "C1-C6 alkoxy" can be understood as "C1-C6 alkyloxy" or "C1-C6 alkyl-O-". The term "C1-C4 alkoxy" can be understood as "C1-C4 alkyloxy" or "C1-C4 alkyl-O-". The "C1-C 10 The term "alkoxy" may include "C1-C6 alkoxy" and "C1-C3 alkoxy", and the "C1-C6 alkoxy" may further include "C1-C3 alkoxy".

[0136] The term "alkenyl" refers to a linear or branched unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms and having at least one double bond. 10 "Alkenyl" is understood to mean a linear or branched unsaturated hydrocarbon radical containing one or more double bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, "C2-C 10 The term "alkenyl" may include "C2-C6 alkenyl", "C2-C4 alkenyl", C2 or C3 alkenyl. It will be understood that when the alkenyl contains more than one double bond, the double bonds may be separated from each other or conjugated. The term "C2-C8 alkenyl" may be understood to mean a straight or branched unsaturated hydrocarbon group containing one or more double bonds and having 2, 3, 4, 5, 6, 7 or 8 carbon atoms. Specific examples of the alkenyl include, but are not limited to, vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl, (E)-1-methylprop-1-enyl or (Z)-1-methylprop-1-enyl, etc.

[0137] The term "alkynyl" refers to a straight or branched unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms and having at least one triple bond. 10 “Alkynyl” is understood to mean a linear or branched unsaturated hydrocarbon radical containing one or more triple bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. “C2-C8 alkynyl” is understood to mean a linear or branched unsaturated hydrocarbon radical containing one or more triple bonds and having 2, 3, 4, 5, 6, 7 or 8 carbon atoms. “C2-C8 10 Examples of "alkynyl" include, but are not limited to, ethynyl (-C≡CH), propynyl (-C≡CCH3, -CH2C≡CH), but-1-ynyl, but-2-ynyl, or but-3-ynyl. "C2-C 10 The term "alkynyl" may include "C2-C3 alkynyl". Examples of "C2-C3 alkynyl" include ethynyl (-C≡CH), prop-1-ynyl (-C≡CCH3), and prop-2-ynyl (-CH2C≡CH).

[0138] The term "cycloalkyl" refers to a fully saturated carbocyclic group that exists in the form of a monocyclic, fused, bridged, or spirocyclic ring. Unless otherwise indicated, the carbocyclic ring is typically a 3- to 20-membered ring. The term "C3-C 12 "Cycloalkyl" refers to a cycloalkyl group having 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 ring carbon atoms. The term "C3-C 10The term "cycloalkyl" refers to a cycloalkyl group having 3, 4, 5, 6, 7, 8, 9 or 10 ring carbon atoms. The term "C3-C6 cycloalkyl" refers to a cycloalkyl group having 3, 4, 5 or 6 ring carbon atoms.

[0139] The term "heterocyclyl" or "heterocycle" refers to a fully saturated or partially saturated (heteroaromatic as a whole that is not aromatic) monocyclic, fused, spiro or bridged ring group, which contains 1-5 (e.g., 1-3 or 1-2) heteroatoms or heteroatomic groups (i.e., heteroatom-containing groups) in its ring atoms, wherein the "heteroatoms or heteroatomic groups" include, but are not limited to, nitrogen atom (N), oxygen atom (O), sulfur atom (S), phosphorus atom (P), boron atom (B), -S(=O)2-, -S(=O)-, -P(=O)2-, -P(=O)-, -NH-, -S(=O)(=NH)-, -C(=O)NH- or -NHC(=O)NH-. The term "4-10 membered heterocyclyl" refers to a heterocyclyl group having 4, 5, 6, 7, 8, 9 or 10 ring atoms, and containing 1-5 heteroatoms or heteroatomic groups independently selected from the above-mentioned heteroatoms or heteroatomic groups in its ring atoms. “4-10 membered heterocyclyl” may include “4-7 membered heterocyclyl”. The term “4-7 membered heterocyclyl” refers to a heterocyclyl having 4, 5, 6 or 7 ring atoms, and containing 1, 2, 3, 4 or 5 heteroatoms or heteroatom groups independently selected from the above-mentioned heteroatoms. Specific examples of 4-membered heterocyclyl include, but are not limited to, azetidinyl or oxetanyl; specific examples of 5-membered heterocyclyl include, but are not limited to, tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, 4,5-dihydrooxazolyl or 2,5-dihydro-1H-pyrrolyl; specific examples of 6-membered heterocyclyl include, but are not limited to, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl, tetrahydropyridinyl or 4H-[1,3,4]thiadiazinyl; specific examples of 7-membered heterocyclyl include, but are not limited to, diazepanyl. The heterocyclic group may also be a bicyclic group, wherein specific examples of 5,5-membered bicyclic groups include but are not limited to hexahydrocyclopenta[c]pyrrol-2(1H)-yl; specific examples of 5,6-membered bicyclic groups include but are not limited to hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazinyl or 5,6,7,8-tetrahydroimidazo[1,5-a]pyrazinyl. Optionally, the heterocyclic group may be a benzo-fused ring group of the above-mentioned 4-7-membered heterocyclic groups, specific examples of which include but are not limited to dihydroisoquinolinyl and the like. The “4-10 membered heterocyclyl” may include the ranges of “5-10 membered heterocyclyl”, “4-7 membered heterocyclyl”, “5-6 membered heterocyclyl”, “6-8 membered heterocyclyl”, “4-10 membered heterocycloalkyl”, “5-10 membered heterocycloalkyl”, “4-7 membered heterocycloalkyl”, “5-6 membered heterocycloalkyl”, “6-8 membered heterocycloalkyl”, and the like, and the “4-7 membered heterocyclyl” may further include the ranges of “4-6 membered heterocyclyl”, “5-6 membered heterocyclyl”, “4-7 membered heterocycloalkyl”, “4-6 membered heterocycloalkyl”, “5-6 membered heterocycloalkyl”, and the like.Although some bicyclic heterocyclic groups disclosed herein partially contain a benzene ring or a heteroaromatic ring, the heterocyclic group as a whole is still non-aromatic.

[0140] The term "heterocycloalkyl" refers to a fully saturated cyclic group in the form of a monocyclic, fused, bridged or spirocyclic ring, wherein the ring atoms of the ring contain 1-5 heteroatoms or heteroatomic groups (i.e., heteroatomic groups containing heteroatoms), wherein the "heteroatoms or heteroatomic groups" include, but are not limited to, nitrogen atom (N), oxygen atom (O), sulfur atom (S), phosphorus atom (P), boron atom (B), -S(=O)2-, -S(=O)-, -NH-, -S(=O)(=NH)-, -C(=O)NH- or -NHC(=O)NH-. The term "4-10 membered heterocycloalkyl" refers to a heterocycloalkyl group having 4, 5, 6, 7, 8, 9 or 10 ring atoms, wherein the ring atoms of the heterocycloalkyl group contain 1-5 heteroatoms or heteroatomic groups independently selected from the above-mentioned heteroatoms or heteroatomic groups. The term "5-10 membered heterocycloalkyl" refers to a heterocycloalkyl group having 5, 6, 7, 8, 9 or 10 ring atoms, and containing 1 to 5 heteroatoms or heteroatom groups independently selected from the above-mentioned heteroatoms. “4-10 membered heterocycloalkyl” and “5-10 membered heterocycloalkyl” include “4-7 membered heterocycloalkyl”, wherein specific examples of 4 membered heterocycloalkyl include but are not limited to azetidinyl, oxetanyl or thietanyl; specific examples of 5 membered heterocycloalkyl include but are not limited to tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, isoxazolidinyl, oxazolidinyl, isothiazolidinyl, thiazolidinyl, imidazolidinyl or tetrahydropyrazolyl; specific examples of 6 membered heterocycloalkyl include but are not limited to piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, piperazinyl, 1,4-thioxanyl, 1,4-dioxanyl, thiomorpholinyl, 1,3-dithianyl or 1,4-dithianyl; specific examples of 7 membered heterocycloalkyl include but are not limited to azepanyl, oxetanyl or thiepanyl.

[0141] The term "aryl" refers to an all-carbon monocyclic or fused polycyclic aromatic ring group having a conjugated π electron system. The aryl group may have 6-20 carbon atoms, 6-14 carbon atoms, or 6-12 carbon atoms. The term "C6-C 10 "Aryl" is understood to mean an aromatic group having 6 to 10 carbon atoms. For example, a ring having 6 carbon atoms ("C6 aryl"), such as phenyl; or a ring having 9 carbon atoms ("C9 aryl"), such as indanyl or indenyl; or a ring having 10 carbon atoms ("C 10 "aryl"), for example tetrahydronaphthyl, dihydronaphthyl or naphthyl.

[0142] The term "heteroaryl" refers to a monocyclic or fused polycyclic ring system having aromatic character, wherein the ring contains at least one ring atom selected from N, O, S, and the remaining ring atoms are C. The term "5-12 membered heteroaryl" is understood to include monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11 or 12 ring atoms, such as 5 or 6 or 9 or 10 or 12 ring atoms, and containing 1 to 5, such as 1 to 3 heteroatoms independently selected from N, O and S. The term "5-10 membered heteroaryl" is understood to include monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8, 9 or 10 ring atoms, such as 5 or 6 or 9 or 10 ring atoms, and containing 1 to 5, such as 1 to 3 heteroatoms independently selected from N, O and S. In particular, the heteroaryl group is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiadiazolyl, and the like, and benzo derivatives thereof, such as benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl or isoindolyl, and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl, and the like, and benzo derivatives thereof, such as quinolyl, quinazolinyl or isoquinolyl, and the like; or acinyl, indolizinyl, purinyl, and the like, and benzo derivatives thereof; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl or phenoxazinyl, and the like. The term "6-10 membered heteroaryl" is understood to include monocyclic or bicyclic aromatic ring systems having 6, 7, 8, 9 or 10 ring atoms, for example 6 or 9 or 10 ring atoms, and containing 1 to 5, for example 1 to 3, heteroatoms independently selected from N, O and S. The term "5-6 membered heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms, and containing 1 to 3, for example 1 to 2, heteroatoms independently selected from N, O and S.

[0143] The term "halo" or "halogen" refers to fluorine, chlorine, bromine or iodine.

[0144] The term "hydroxy" refers to an -OH group.

[0145] The term "cyano" refers to a -CN group.

[0146] The term "amino" refers to a -NH2 group.

[0147] The term "nitro" refers to a -NO2 group.

[0148] The term "mercapto" refers to a -SH group.

[0149] The term "treatment" means administering the compound or formulation described herein to improve or eliminate a disease or one or more symptoms associated with the disease, and includes:

[0150] (i) inhibiting a disease or disease state, i.e., arresting its development;

[0151] (ii) ameliorating the disease or condition, i.e., causing regression of the disease or condition.

[0152] The term "therapeutically effective amount" means an amount of a compound of the present disclosure that (i) treats a particular disease, condition, or disorder, and (ii) alleviates, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder. The amount of a compound of the present disclosure that constitutes a "therapeutically effective amount" varies depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by one skilled in the art based on their knowledge and this disclosure.

[0153] The term "prevention" means administering a compound or formulation described herein to prevent a disease or one or more symptoms associated with the disease, and includes preventing the disease or disease state from occurring in an individual (e.g., a mammal), particularly when such individual (e.g., a mammal) is susceptible to the disease state but has not yet been diagnosed as having the disease state.

[0154] The term "subject" includes mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class Mammalia: humans, non-human primates (e.g., chimpanzees and other apes and monkeys); livestock, such as cattle, horses, sheep, goats, and pigs; domestic animals, such as rabbits, dogs, and cats; and laboratory animals, including rodents, such as rats, mice, and guinea pigs. Examples of non-human mammals include, but are not limited to, birds and fish. In one embodiment of the methods and compositions provided herein, the mammal is a human. The terms "patient" and "subject" are used interchangeably.

[0155] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0156] The term "pharmaceutically acceptable salt" refers to a salt of a pharmaceutically acceptable acid or base, including a salt formed between a compound and an inorganic acid or organic acid, and a salt formed between a compound and an inorganic base or an organic base.

[0157] The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present disclosure or their salts and a pharmaceutically acceptable excipient. The purpose of a pharmaceutical composition is to facilitate administration of the compounds of the present disclosure to an organism.

[0158] The term "pharmaceutically acceptable excipient" refers to an excipient that is non-irritating to organisms and does not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art and include, for example, carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.

[0159] The word "comprise" or "comprises" and its English variations such as comprises or comprising are to be understood as having an open and non-exclusive meaning, ie, "including but not limited to".

[0160] The present disclosure also includes isotopically labeled compounds of the present disclosure that are identical to those described herein, but where one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2 H. 3 H. 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 123 I. 125 I and 36 Cl et al.

[0161] Certain isotopically labeled compounds of the present disclosure (e.g., 3 H and 14 C-labeled) can be used in compound and / or substrate tissue distribution assays. 3 H) and carbon-14 (i.e. 14 C) isotopes are particularly preferred due to their ease of preparation and detectability. Positron emitting isotopes, such as 15 O. 13 N. 11 C and 18 F can be used in positron emission tomography (PET) studies to determine substrate occupancy. Isotopically labeled compounds of the disclosure can generally be prepared by following procedures analogous to those disclosed in the Schemes and / or Examples below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0162] The pharmaceutical compositions of the present disclosure can be prepared by combining the compounds of the present disclosure with suitable pharmaceutically acceptable excipients, and can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols.

[0163] Typical routes of administration of the disclosed compounds, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0164] The pharmaceutical composition of the present disclosure can be manufactured by methods well known in the art, such as conventional mixing methods, dissolution methods, granulation methods, emulsification methods, freeze-drying methods, and the like.

[0165] In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical composition can be formulated by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present disclosure to be formulated into tablets, pills, lozenges, dragees, capsules, liquids, gels, slurries, suspensions, and the like for oral administration to a patient.

[0166] Solid oral compositions can be prepared by conventional mixing, filling, or tableting methods. For example, they can be prepared by mixing the active compound with a solid excipient, optionally grinding the resulting mixture, adding other suitable excipients as needed, and then granulating the mixture to obtain a tablet or dragee core. Suitable excipients include, but are not limited to, binders, diluents, disintegrants, lubricants, glidants, or flavoring agents.

[0167] The pharmaceutical composition may also be suitable for parenteral administration, such as sterile solutions, suspensions or lyophilized products in appropriate unit dosage forms.

[0168] The dosage administered will depend on factors such as the specific compound, the disease state and its severity, the identity of the subject or host to be treated (e.g., weight, sex), and will be determined by the particular circumstances of the case, including, for example, the specific formulation being administered, the route of administration, the condition being treated, and the subject or host being treated.

[0169] In all methods of administration of the compounds of formula (I) described herein, oral administration is administered at a daily dose of 0.001 mg / kg to 5000 mg / kg body weight, preferably 0.01 mg / kg to 100 mg / kg body weight, in single or divided doses. The daily dose and unit dose vary depending on many variables, including but not limited to the activity of the compound used, the disease or condition to be treated, the mode of administration, the requirements of the individual subject, the severity of the disease or condition to be treated, and the judgment of the practitioner.

[0170] The compounds disclosed herein can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining the same with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include, but are not limited to, the examples disclosed herein.

[0171] The chemical reactions of the disclosed embodiments are carried out in suitable solvents that are compatible with the chemical transformations disclosed herein and the reagents and materials required. To obtain the compounds disclosed herein, it may sometimes be necessary for those skilled in the art to modify or select synthetic steps or reaction schemes based on existing embodiments.

[0172] This disclosure uses the following abbreviations:

[0173] EtOH represents ethanol; Triethyl orthoformate represents triethyl orthoformate; 1,4-dioxane / dioxane represents 1,4-dioxane; B2Pin2 represents diboronic acid pinacol ester; KOAc represents potassium acetate; Pd(dppf)Cl2 represents [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride; Pd(PPh3)4 represents tetrakis(triphenylphosphine)palladium; Pd2(dba)3: tris(dibenzylideneacetone)dipalladium; xantphos represents 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene; MeCN represents acetonitrile; TFA represents trifluoroacetic acid; DMF-DMA represents N,N-dimethylformamide dimethyl acetal; THF represents tetrahydrofuran; TFAA represents trifluoroacetic anhydride; n-Butyllithium represents n-butyllithium; [Rh(Cp*)Cl2]2 represents dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer; Trimethoxymethane represents trimethyl orthoformate; Ms2O represents methanesulfonic anhydride; TEA represents triethylamine; DCM represents dichloromethane; DMF represents N,N-dimethylformamide; DMB represents 2,4-dimethoxybenzyl; DMBNH2 represents (2,4-dimethoxyphenyl)methylamine; 4A MS stands for 4A molecular sieve; IPA / iPrOH stands for isopropyl alcohol; iPrMgCl stands for isopropyl magnesium chloride; 2,4,6-Collidine stands for 2,4,6-trimethylpyridine; TCFH stands for N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate; Bu stands for butyl; IBX stands for 2-iodoacylbenzoic acid; TsNHNH2 stands for p-toluenesulfonylhydrazide; Pd(dtbpf)Cl2 stands for [1,1'-bis(di-tert-butylphosphino)ferrocene]palladium dichloride; DMSO stands for dimethyl sulfoxide; LC-MS stands for liquid chromatography-mass spectrometry; MS stands for mass spectrometry; 1 H NMR stands for proton nuclear magnetic resonance; ESI stands for electrospray ionization; HEPES stands for N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid buffer; EGTA stands for ethylene glycol-bis(2-aminoethyl ether)tetraacetic acid; DTT stands for dithiothreitol; ATP stands for adenosine triphosphate; EDTA stands for ethylenediaminetetraacetic acid; IC 50 50% inhibitory concentration, which refers to the concentration that achieves half of the maximal inhibitory effect; ELISA stands for enzyme-linked immunosorbent assay; FBS stands for fetal bovine serum.

[0174] DETAILED DESCRIPTION

[0175] The compounds of the present disclosure can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed herein, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present disclosure.

[0176] The present disclosure is described in detail below by way of examples, but this is not intended to limit the present disclosure in any way. While the present disclosure has been described in detail herein, including specific embodiments thereof, it will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments without departing from the spirit and scope of the present disclosure. All reagents used in the present disclosure are commercially available and can be used without further purification.

[0177] Unless otherwise specified, the ratios expressed for mixed solvents are volume ratios.

[0178] Unless otherwise specified, % refers to weight %.

[0179] Compounds are manually or Software naming, commercially available compounds use supplier catalog names.

[0180] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The units of NMR shifts are 10 -6 (ppm). The solvents for NMR measurements are deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard is tetramethylsilane (TMS);

[0181] The eluent or mobile phase may be a mixed eluent or mobile phase composed of two or more solvents, wherein the ratio is the volume ratio of each solvent. Example 1: (3S,4R)-4-((5-fluoro-4-(5-methyl-[1,2,4]triazolo[3,4-a]isoquinolin-8-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 1)

[0182] Step 1: 6-Bromo-1-chloro-3-methylisoquinoline (Compound 1-2)

[0183] Dissolve 6-bromo-3-methylisoquinolin-1(2H)-one (Compound 1-1, 100.0 mg, 0.42 mmol) in ultra-dry acetonitrile (2 mL). Add phosphorus oxychloride (193.0 mg, 1.26 mmol). Heat at 80°C under nitrogen for 2 hours to complete the reaction. Concentrate the reaction system to dryness under reduced pressure to obtain the crude title compound 1-2 (120.0 mg).

[0184] LC-MS: m / z(ESI):256[M+H] + .

[0185] Step 2: 6-Bromo-1-hydrazino-3-methylisoquinoline (Compound 1-3)

[0186] The crude compound 1-2 (120.0 mg) was dissolved in ethanol (2 mL), and then hydrazine hydrate (80%, 0.5 mL) was added and heated at 80°C for 16 hours. The reaction was completed. The reaction solution was directly concentrated under reduced pressure to obtain the crude title compound 1-3 (200.0 mg).

[0187] LC-MS: m / z(ESI):252[M+H] + .

[0188] Step 3: 8-Bromo-5-methyl-[1,2,4]triazolo[3,4-a]isoquinoline (Compound 1-4)

[0189] The crude product compound 1-3 (200.0 mg) was dissolved in triethyl orthoformate (2 mL) and heated at 80°C under a nitrogen atmosphere for 16 hours to complete the reaction. The reaction system was added to water and extracted three times with ethyl acetate. The organic phases were combined and backwashed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure. The filtrate was purified by reverse-phase column chromatography (water (containing 0.1% formic acid): acetonitrile = 1:1) to obtain the title compound 1-4 (55 mg, 50% yield over three steps).

[0190] LC-MS: m / z(ESI):262[M+H] + .

[0191] Step 4: 5-methyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,2,4]triazolo[3,4-a]isoquinoline (Compound 1-5)

[0192] Compound 1-4 (55 mg, 0.21 mmol) was dissolved in 1,4-dioxane (1 mL), followed by the addition of [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (17 mg, 0.02 mmol), bipyraclostrobin (106.6 mg, 0.42 mmol), and potassium acetate (75.0 mg, 0.42 mmol). The mixture was heated at 80°C under nitrogen for 2 hours to complete the reaction. The reaction mixture was filtered and the filtrate was concentrated to dryness under reduced pressure to obtain the crude title compound 1-5 (60 mg).

[0193] LC-MS: m / z(ESI):310[M+H] + .

[0194] Step 5: 8-(2-chloro-5-fluoropyrimidin-4-yl)-5-methyl-[1,2,4]triazolo[3,4-a]isoquinoline (Compound 1-7)

[0195] Compound 1-5 (60 mg) was dissolved in a mixture of 1,4-dioxane (1 mL) and water (0.25 mL). Tetrakis(triphenylphosphine)palladium (22 mg, 0.02 mmol), 2,4-dichloro-5-fluoropyrimidine (compound 1-6, 48.6 mg, 0.29 mmol), and sodium carbonate (41.0 mg, 0.39 mmol) were then added. The mixture was heated at 90°C under a nitrogen atmosphere for 16 hours to complete the reaction. The reaction solution was directly filtered, and the filtrate was concentrated to dryness under reduced pressure. The title compound 1-7 (30 mg, 45% yield over two steps) was obtained by purification by normal phase column chromatography (dichloromethane:methanol = 20:1-10:1).

[0196] LC-MS: m / z(ESI):314[M+H] + .

[0197] Step 6: (3S,4R)-4-((5-fluoro-4-(5-methyl-[1,2,4]triazolo[3,4-a]isoquinolin-8-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 1)

[0198] Compound 1-7 (30.0 mg, 0.095 mmol) was dissolved in 1,4-dioxane (1 mL), followed by the addition of tris(dibenzylideneacetone)dipalladium (8.7 mg, 0.01 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (11.0 mg, 0.02 mmol), (3S,4R)-4-aminotetrahydro-2H-pyran-3-ol (compound 1-8, 16.8 mg, 0.14 mmol), and cesium carbonate (62.0 mg, 0.20 mmol). The reaction was heated at 95°C under nitrogen for 16 hours, completing the reaction. The reaction solution was filtered, and the filtrate was concentrated to dryness under reduced pressure. The residue was purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:1) to obtain compound 1 (11.0 mg, 29% yield).

[0199] LC-MS: m / z(ESI):395[M+H] + .

[0200] 1H NMR (400MHz, DMSO-d6) δ9.40(s,1H),8.66(d,J=8.0Hz,1H),8.50(d,J=4.0Hz,1H),8.43(s,1H),8.27(d,J=8.0Hz,1H),7.28-7.26(m,2H),4.93( d,J=4.0Hz,1H),3.85-3.81(m,3H),3.54-3.51(m,1H),3.41-3.35(m,1H ),3.11-3.06(m,1H),2.72(s,3H),2.03-1.99(m,1H),1.55-1.46(m,1H).

[0201] Example 2: (3S,4R)-4-((5-fluoro-4-(4-methylpyrazolo[1,5-a]quinolin-7-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 2)

[0202] Step 1: 1-amino-6-bromo-3-methylquinolin-1-ium-2,4-dinitrophenolate (Compound 2-3)

[0203] Compound 2-1 (200.0 mg, 0.9 mmol) was dissolved in acetonitrile (2 mL), and compound 2-2 (219.0 mg, 1.1 mmol) was added. The mixture was reacted at room temperature for 2 hours. After the reaction was completed, the crude compound 2-3 (300.0 mg) was obtained by filtration.

[0204] LC-MS:m / z(ESI):237[M-C6H3N2O5] + .

[0205] Step 2: 7-Bromo-4-methylpyrazolo[1,5-a]quinoline (Compound 2-5)

[0206] The crude compound 2-3 (300.0 mg) was dissolved in acetonitrile (2 mL), and then compound 2-4 (612 mg, 7.1 mmol) was added. The reaction was heated at 60°C for 16 hours to complete the reaction. The reaction solution was directly concentrated to dryness under reduced pressure and purified by normal phase column chromatography (petroleum ether:ethyl acetate = 20:1-1:1) to obtain compound 2-5 (10 mg).

[0207] LC-MS: m / z(ESI):261[M+H] + .

[0208] Step 3: 4-methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[1,5-a]quinoline (Compound 2-6)

[0209] Compound 2-5 (15.0 mg, 0.057 mmol) was dissolved in 1,4-dioxane (1 mL), followed by the addition of [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (4.5 mg, 0.006 mmol), bipyralidoboric acid pinacol ester (21.6 mg, 0.086 mmol), and potassium acetate (16.9 mg, 0.17 mmol). The mixture was heated at 80°C under nitrogen for 2 hours to complete the reaction. The reaction solution was directly filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain crude compound 2-6 (18 mg).

[0210] LC-MS: m / z(ESI):309[M+H] + .

[0211] Step 4: (3S,4R)-4-((5-fluoro-4-(4-methylpyrazolo[1,5-a]quinolin-7-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-yl acetate (Compound 2-8)

[0212] Compound 2-6 (18 mg) was dissolved in a mixture of 1,4-dioxane (1 mL) and water (0.1 mL). Tetrakis(triphenylphosphine)palladium (7 mg, 0.006 mmol), compound 2-7 (33 mg, 0.116 mmol), and sodium carbonate (18.5 mg, 0.17 mmol) were then added. The mixture was heated at 90°C under a nitrogen atmosphere for 16 hours to complete the reaction. The reaction solution was concentrated to dryness under reduced pressure and purified by reverse phase column chromatography (eluent: water:acetonitrile = 1:1) to obtain compound 2-8 (15 mg, 59% yield over two steps).

[0213] LC-MS: m / z(ESI):436[M+H] + .

[0214] Step 6: (3S,4R)-4-((5-fluoro-4-(4-methylpyrazolo[1,5-a]quinolin-7-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 2)

[0215] Compound 2-8 (15.0 mg, 0.034 mmol) was dissolved in a mixture of methanol (1 mL) and water (1 mL), followed by the addition of potassium carbonate (14.0 mg, 0.1 mmol). The mixture was heated at 50°C for 1 hour to complete the reaction. The reaction solution was purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:1) to obtain compound 2 (5.0 mg, yield 37%).

[0216] LC-MS: m / z(ESI):394[M+H] + .

[0217] 1H NMR(400MHz, DMSO-d6)δ8.65(s,2H),8.55(d,J=8.9Hz,1H),8.51(d,J=3.6Hz,1H),8.43–8.37(m,1H),8.07(s,1H),7.31–7.24(m,1H) ,4.95(d,J=5.4Hz,1H),3.89–3.80(m,3H),3.56–3.49(m,2H),3.14–3.05(m,1H),2.64(s,3H),2.03–1.95(m,1H),1.52–1.42(m,1H).

[0218] Example 3: (3S,4R)-4-((5-fluoro-4-(4-methyl-[1,2,4]triazolo[1,5-a]quinolin-7-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 3)

[0219] Step 1: 6-Bromo-N-(2,4-dimethoxybenzyl)-3-methylquinolin-2-amine (Compound 3-2)

[0220] Compound 3-1 (700.0 mg, 2.73 mmol) and (2,4-dimethoxyphenyl)methylamine (912 mg, 5.46 mmol) were dissolved in dimethyl sulfoxide (7 mL), potassium carbonate (754.0 mg, 5.46 mmol) was added, and the reaction was heated at 80°C under nitrogen for 40 hours to complete the reaction. Saturated aqueous ammonium chloride solution was added, and the mixture was extracted with ethyl acetate three times. The organic phase was backwashed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. The mixture was purified by normal phase column chromatography (petroleum ether:ethyl acetate = 10:1-1:1) to obtain compound 3-2 (400 mg, yield 38%).

[0221] LC-MS: m / z(ESI):387[M+H] + .

[0222] Step 2: 6-Bromo-3-methylquinolin-2-amine (Compound 3-3)

[0223] Compound 3-2 (100.0 mg) was dissolved in trifluoroacetic acid (2 mL) and reacted at room temperature for 1 hour. The reaction was completed. The reaction solution was directly concentrated to dryness under reduced pressure, added with saturated sodium bicarbonate aqueous solution, extracted three times with ethyl acetate, and the organic phase was backwashed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain crude compound 3-3 (70.0 mg).

[0224] LC-MS: m / z(ESI):237[M+H] + .

[0225] Step 3: (Z)-N-(6-bromo-3-methylquinolin-2-yl)-N'-hydroxycarboxamidine (Compound 3-4)

[0226] Dissolve the crude compound 3-3 (70.0 mg, 0.30 mmol) in isopropanol (2 mL), add N,N-dimethylformamide dimethyl acetal (180 mg, 1.52 mmol), and heat to 80°C for 1 hour under nitrogen. Cool to 50°C, add hydroxylamine hydrochloride (121 mg, 1.77 mmol), and heat to 50°C for 1 hour to complete the reaction. The reaction solution was concentrated to dryness under reduced pressure to obtain crude compound 3-4 (300 mg).

[0227] LC-MS: m / z(ESI):280[M+H] + .

[0228] Step 4: 7-Bromo-4-methyl-[1,2,4]triazolo[1,5-a]quinoline (Compound 3-5)

[0229] The crude product compound 3-4 (300.0 mg, 1.07 mmol) was dissolved in tetrahydrofuran (2 mL), and trifluoroacetic anhydride (316 mg, 1.5 mmol) was added. The reaction was allowed to react at room temperature for 1 hour. A saturated aqueous sodium bicarbonate solution was added, and the mixture was extracted three times with ethyl acetate. The organic phase was backwashed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. Purification by normal phase column chromatography (petroleum ether:ethyl acetate = 10:1 to 1:1) gave the title compound 3-5 (40 mg, 60% yield over three steps).

[0230] LC-MS: m / z(ESI):262[M+H] + .

[0231] Step 5: 4-methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,2,4]triazolo[1,5-a]quinoline (Compound 3-6)

[0232] Compound 3-5 (10 mg, 0.038 mmol) was dissolved in 1,4-dioxane (1 mL), followed by the addition of [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (3 mg, 0.004 mmol), bipyraclostrobin (14.5 mg, 0.057 mmol), and potassium acetate (11.0 mg, 0.114 mmol). The mixture was heated at 80°C under nitrogen for 2 hours to complete the reaction. The reaction mixture was filtered and the filtrate was concentrated to dryness under reduced pressure to obtain crude compound 3-6 (12 mg).

[0233] LC-MS: m / z(ESI):310[M+H]+ .

[0234] Step 6: (3S,4R)-4-((5-fluoro-4-(4-methyl-[1,2,4]triazolo[1,5-a]quinolin-7-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-yl acetate (Compound 3-7)

[0235] Compound 3-6 (12 mg) was dissolved in a mixture of 1,4-dioxane (1 mL) and water (0.1 mL). Tetrakis(triphenylphosphine)palladium (4.7 mg, 0.004 mmol), compound 2-7 (22 mg, 0.076 mmol), and sodium carbonate (12.1 mg, 0.115 mmol) were then added. The mixture was heated at 90°C under a nitrogen atmosphere for 16 hours to complete the reaction. The reaction solution was concentrated to dryness under reduced pressure and purified by reverse phase column chromatography (eluent: water:acetonitrile = 1:1) to obtain compound 3-7 (10 mg, 60% yield over two steps).

[0236] LC-MS: m / z(ESI):437[M+H] + .

[0237] Step 7: (3S,4R)-4-((5-fluoro-4-(4-methyl-[1,2,4]triazolo[1,5-a]quinolin-7-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 3)

[0238] Compound 3-7 (20.0 mg, 0.045 mmol) was dissolved in a mixture of methanol (1 mL) and water (1 mL), and potassium carbonate (14.0 mg, 0.1 mmol) was added. The mixture was heated at 50°C for 1 hour to complete the reaction. The reaction solution was purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:1) to obtain compound 3 (5.0 mg, yield 27%).

[0239] LC-MS: m / z(ESI):395[M+H] + .

[0240] 1H NMR (400MHz, DMSO-d6) δ9.40(s,1H),8.66(d,J=8.0Hz,1H),8.50(d,J=4.0Hz,1H),8.43(s,1H),8.27(d,J=8.0Hz,1H),7.28-7.26(m,2H),4.93( d,J=4.0Hz,1H),3.85-3.81(m,3H),3.54-3.51(m,1H),3.41-3.35(m,1H ),3.11-3.06(m,1H),2.72(s,3H),2.03-1.99(m,1H),1.55-1.46(m,1H).

[0241] Example 4: (3S,4R)-4-((4-(5-(aminomethyl)-6-isopropyl-[1,2,4]triazolo[3,4-a]isoquinolin-8-yl)-5-fluoropyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 4)

[0242] Step 1: Benzyl 4-methylpent-2-ynoate (Compound 4-2)

[0243] The air in a dry flask was replaced with argon, and tetrahydrofuran (8 mL) and 3-methyl-1-butyne (1.1 g, 16.1 mmol) were added to the flask in sequence. After the reaction solution was cooled to -78°C, n-butyllithium solution (19.3 mmol, 1.6 mmol / mL) was slowly added dropwise to the reaction solution. The reaction solution was stirred at low temperature for ten minutes. The reaction solution was allowed to warm to room temperature naturally and reacted for half an hour to prepare an alkynyl lithium solution. In another flask, the air was replaced with argon, and tetrahydrofuran (10 mL) and compound 4-1 (3.57 g, 21.0 mmol) were added to the flask. After the reaction solution was cooled to -78°C, the alkynyl lithium solution was slowly added dropwise to the reaction solution. The reaction solution was stirred at low temperature for ten minutes. The reaction solution was allowed to warm to room temperature naturally and reacted for 1 hour. After the reaction, saturated ammonium chloride solution was added to the mixture in an ice bath to quench the reaction. Ethyl acetate and water were added for extraction. The organic layer was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by normal phase column chromatography (petroleum ether:ethyl acetate = 5:1) to obtain compound 4-2 (2.8 g, yield 66%). m / z (ESI): 203 [M+H] + .

[0244] Step 2: Benzyl 6-bromo-4-isopropyl-1-oxo-1,2-dihydroisoquinoline-3-carboxylate (Compound 4-4)

[0245] Compound 4-3 (3.45 g, 11.5 mmol), dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer (35.5 mg, 0.058 mmol), and cesium acetate (2.2 g, 11.5 mmol) were dissolved in methanol (15 mL). The reaction solution was stirred at room temperature, and then compound 4-2 (2.8 g, 13.8 mmol) was added to the reaction solution. The reaction was continued at room temperature for 16 hours. After completion of the reaction, the reaction solution was concentrated and purified by normal phase column chromatography (petroleum ether:ethyl acetate = 5:1) to obtain compound 4-4 (3.45 g, 75% yield).

[0246] m / z(ESI):400[M+H] + .

[0247] Step 3: Benzyl 6-bromo-1-chloro-4-isopropylisoquinoline-3-carboxylate (Compound 4-5)

[0248] Compound 4-4 (0.9 g, 2.3 mmol) was placed in a reaction tube. The air in the reaction tube was replaced with argon. Anhydrous tetrahydrofuran (10 mL) was added to the reaction tube and stirred at room temperature. Phosphorus oxychloride (1.0 g, 6.7 mmol) was then slowly added dropwise. After the addition was complete, the reaction solution was stirred in a 90°C oil bath for 3 hours. After the reaction was completed, the reaction tube was cooled in an ice bath. After cooling, triethylamine (9.3 g, 92 mmol) was slowly added to neutralize the reaction. The mixture was then purified by normal phase column chromatography (petroleum ether:ethyl acetate = 5:1) to obtain compound 4-5 (0.55 g, 57% yield).

[0249] m / z(ESI):418[M+H] + .

[0250] Step 4: (6-Bromo-1-chloro-4-isopropylisoquinolin-3-yl)methanol (Compound 4-6)

[0251] Compound 4-5 (102.0 mg, 0.24 mmol) and sodium borohydride (20.0 mg, 0.53 mmol) were placed in a reaction tube. The air in the reaction tube was replaced with argon, and tetrahydrofuran (5 mL) and methanol (0.1 mL) were added to the reaction tube. The reaction solution was stirred at room temperature for 1 hour and then stirred in a 65°C oil bath for 6 hours. After the reaction, the reaction solution was concentrated and purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:9) to obtain compound 4-6 (50.0 mg, yield 66%).

[0252] m / z(ESI):314[M+H] + .

[0253] Step 5: (6-Bromo-1-hydrazino-4-isopropylisoquinolin-3-yl)methanol (Compound 4-7)

[0254] Compound 4-6 (50.0 mg, 0.16 mmol) was placed in a reaction tube. The air in the reaction tube was replaced with argon. Water, hydrazine (1 mL), and ethanol (2 mL) were added to the reaction tube. The reaction solution was stirred in a 90°C oil bath for 8 hours. After the reaction, the reaction solution was concentrated and purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:9) to obtain compound 4-7 (40.0 mg, yield 81%).

[0255] m / z(ESI):310[M+H] + .

[0256] Step 6: (8-Bromo-6-isopropyl-[1,2,4]triazolo[3,4-a]isoquinolin-5-yl)methanol (Compound 4-8)

[0257] Compound 4-7 (40.0 mg, 0.13 mmol) was placed in a reaction tube, the atmosphere was replaced with argon, and trimethyl orthoformate (2 mL) was added. The reaction mixture was stirred in a 90°C oil bath for 8 hours. After completion of the reaction, the reaction mixture was concentrated and purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:9) to obtain compound 4-8 (30.0 mg, 73% yield).

[0258] m / z(ESI):320[M+H] + .

[0259] Step 7: 2-((8-Bromo-6-isopropyl-[1,2,4]triazolo[3,4-a]isoquinolin-5-yl)methyl)isoindoline-1,3-dione (Compound 4-9)

[0260] Compound 4-8 (40 mg, 0.13 mmol) and methanesulfonic anhydride (34.0 mg, 0.19 mmol) were placed in a dry reaction tube. The air in the reaction tube was replaced with argon, and dichloromethane (2 mL) was added to the reaction tube. The reaction solution was stirred in an ice-water bath. Triethylamine (39.0 mg, 0.39 mmol) was added dropwise to the reaction solution. After the addition was complete, the reaction solution was stirred at room temperature for 3 hours. After the reaction was completed, the reaction solution was concentrated and dried under vacuum to obtain the intermediate. Potassium phthalimide (48 mg, 0.26 mmol) was added to the reaction tube. The air in the reaction tube was replaced with argon, and N,N-dimethylformamide (2 mL) was added to the reaction tube. The reaction solution was stirred in an 80°C oil bath for 6 hours. After the reaction was completed, the reaction solution was purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:9) to obtain compound 4-9 (15.0 mg, yield 26%).

[0261] m / z(ESI):449[M+H] + .

[0262] Step 8: 2-((6-isopropyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,2,4]triazolo[3,4-a]isoquinolin-5-yl)methyl)isoindoline-1,3-dione (Compound 4-10)

[0263] Compound 4-9 (15.0 mg, 0.033 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (2.4 mg, 0.0033 mmol), pinacol diboron (12.6 mg, 0.05 mmol), and potassium acetate (9.8 mg, 0.1 mmol) were placed in a reaction tube. The air in the reaction tube was replaced with argon. 1,4-dioxane (2 mL) was added, and the reaction solution was stirred in a 90°C oil bath for 10 hours. After the reaction was completed, the reaction solution was cooled to room temperature and filtered to obtain crude compound 4-10 (16.8 mg).

[0264] m / z(ESI):497[M+H] + .

[0265] Step 9: 2-(((8-(2-(((3S,4R)-3-acetoxytetrahydro-2H-pyran-4-yl)amino)-5-fluoropyrimidin-4-yl)-6-isopropyl-[1,2,4]triazolo[3,4-a]isoquinolin-5-yl)methyl)carbamoyl)benzoic acid (Compound 4-11)

[0266] The crude product, compound 4-10 (16.8 mg, 0.034 mmol), compound 2-7 (11.6 mg, 0.04 mmol), sodium carbonate (10.6 mg, 0.1 mmol), tetrakis(triphenylphosphine)palladium (3.8 mg, 0.0033 mmol), 1,4-dioxane (2 mL), and water (0.1 mL) were placed in a reaction tube. The air in the reaction tube was replaced with argon. The reaction solution was stirred in a 100°C oil bath for 8 hours. After the reaction, the reaction solution was concentrated and purified by reverse phase column chromatography (eluent: water:acetonitrile = 1:9) to obtain compound 4-11 (5.0 mg, yield 23%).

[0267] m / z(ESI):642[M+H] + .

[0268] Step 10: (3S,4R)-4-((4-(5-(aminomethyl)-6-isopropyl-[1,2,4]triazolo[3,4-a]isoquinolin-8-yl)-5-fluoropyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 4)

[0269] Compound 4-11 (5.0 mg, 0.0078 mmol) and ethanol (1 mL) were placed in a reaction tube. The air in the reaction tube was replaced with argon. Hydrazine hydrate (0.5 mL) was added to the reaction tube, and the reaction tube was stirred in a 90°C oil bath for 8 hours. After the reaction, the reaction solution was purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:9) to obtain compound 4 (2.5 mg, yield 71%).

[0270] m / z(ESI):452[M+H] + .

[0271] 1 H NMR (400MHz, DMSO-d6) δ9.45 (s, 1H), 8.97 (s, 1H), 8.74 (d, J = 8.4Hz, 1H), 8.52 (d, J=3.8Hz,1H),8.30(d,J=8.4Hz,1H),7.28(d,J=7.8Hz,1H),4.96(d,J=5.3Hz,1H), 4.25(s,2H),3.92–3.82(m,4H),3.55(tt,J=9.6,4.9Hz,1H),3.40–3.35(m,2H),3. 12–3.01(m,1H),2.18(s,2H),2.04(d,J=13.3Hz,1H),1.57(dd,J=7.2,4.8Hz,6H).

[0272] Example 5: (R)-1-((8-(5-fluoro-2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)-6-isopropyl-[1,2,4]triazolo[3,4-a]isoquinolin-5-yl)methyl)piperidine-3-carbonitrile (Compound 5)

[0273] Step 1: (R)-1-((8-bromo-6-isopropyl-[1,2,4]triazolo[3,4-a]isoquinolin-5-yl)methyl)piperidine-3-carbonitrile (Compound 5-2)

[0274] Compound 4-8 (22 mg, 0.069 mmol) and methanesulfonic anhydride (24.0 mg, 0.14 mmol) were placed in a dry reaction tube, the air in the reaction tube was replaced with argon, dichloromethane (2 mL) was added to the reaction tube, the reaction solution was placed in an ice bath and stirred, triethylamine (20.0 mg, 0.2 mmol) was added dropwise to the reaction solution, and after completion of the addition, the mixture was stirred at room temperature for 3 hours. After the reaction, the reaction solution was concentrated and dried in vacuo to obtain the intermediate; another dry reaction tube was added with compound 5-1 (29.0 mg, 0.14 mmol) and ethyl acetate hydrochloride (1 mL, 4 mol / L) solution, and stirred at room temperature for 2 hours. After the reaction was completed, the solid was concentrated to form a solid, which was placed in a reaction tube with the aforementioned reaction intermediate. The air in the reaction tube was replaced with argon, and N,N-dimethylformamide (2 mL) and triethylamine (20.0 mg, 0.2 mmol) were added to the reaction tube. The reaction solution was placed in a 50°C oil bath and stirred for 6 hours. After the reaction, the reaction solution was purified by reverse phase column chromatography (eluent: water:acetonitrile = 1:9) to obtain compound 5-2 (12.0 mg, yield 42%).

[0275] m / z(ESI):412[M+H] + .

[0276] Step 2: (R)-1-((6-isopropyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,2,4]triazolo[3,4-a]isoquinolin-5-yl)methyl)piperidine-3-carbonitrile (Compound 5-3)

[0277] Compound 5-2 (12.0 mg, 0.029 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (2.1 mg, 0.0029 mmol), pinacol diboron (9.6 mg, 0.038 mmol), and potassium acetate (8.5 mg, 0.087 mmol) were placed in a reaction tube. The air in the reaction tube was replaced with argon. 1,4-dioxane (2 mL) was added, and the reaction solution was stirred in a 90°C oil bath for 10 hours. After the reaction, the reaction solution was cooled to room temperature and filtered to obtain a dioxane solution of crude compound 5-3 for later use.

[0278] m / z(ESI):460[M+H] + .

[0279] Step 3: (3S,4R)-4-((4-(5-(((R)-3-cyanopiperidin-1-yl)methyl)-6-isopropyl-[1,2,4]triazolo[3,4-a]isoquinolin-8-yl)-5-fluoropyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-yl acetate (Compound 5-4)

[0280] A dioxane solution of the crude compound 5-3, sodium carbonate (9.2 mg, 0.087 mmol), compound 2-7 (11.6 mg, 0.04 mmol), tetrakis(triphenylphosphine)palladium (3.3 mg, 0.0029 mmol), and water (0.1 mL) were placed in a reaction tube. The air in the reaction tube was replaced with argon. The reaction solution was stirred in a 100°C oil bath for 8 hours. After completion of the reaction, the reaction solution was concentrated and purified by reverse phase column chromatography (eluent: water:acetonitrile = 1:9) to obtain compound 5-4 (6.5 mg, 38% yield).

[0281] m / z(ESI):587[M+H] + .

[0282] Step 4: (R)-1-((8-(5-fluoro-2-(((3S,4R)-3-hydroxytetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)-6-isopropyl-[1,2,4]triazolo[3,4-a]isoquinolin-5-yl)methyl)piperidine-3-carbonitrile (Compound 5)

[0283] Compound 5-4 (6.5 mg, 0.011 mmol) and ethanol (1 mL) were placed in a reaction tube. The air in the reaction tube was replaced with argon. Hydrazine hydrate (0.5 mL) was added to the reaction tube, and the reaction tube was stirred in a 60°C oil bath for 6-8 hours. After the reaction, the reaction solution was purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:9) to obtain compound 5 (4.2 mg, 70% yield).

[0284] m / z(ESI):545[M+H] + .

[0285] 1 H NMR (400MHz, DMSO-d6) δ9.35 (s, 1H), 9.03 (s, 1H), 8.76 (d, J = 8.4Hz, 1H), 8.52 (d, J = 3 .8Hz,1H),8.33(dt,J=8.4,1.4Hz,1H),7.30(d,J=7.9Hz,1H),4.99(s,1H),4.10(s,2 H),3.95–3.81(m,4H),3.38(d,J=2.2Hz,1H),3.09–3.01(m,2H),2.80(d,J=27.3Hz,2 H),2.60(s,1H),2.45(s,3H),2.04(d,J=13.1Hz,1H),1.69(s,2H),1.63–1.46(m,8H).

[0286] Example 6: (3S,4R)-4-((4-(5-((3,3-difluoropiperidin-1-yl)methyl)-6-isopropyl-[1,2,4]triazolo[3,4-a]isoquinolin-8-yl)-5-fluoropyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 6)

[0287] According to the synthesis method of compound 5, the compound 5-1 in the first step is replaced by (tert-Butyl 3,3-difluoropiperidine-1-carboxylate) gave compound 6 (6.6 mg, yield 79%).

[0288] m / z(ESI):556[M+H] + .

[0289] 1 H NMR (400MHz, DMSO-d6) δ9.31(s,1H),9.03(s,1H),8.76(d,J=8.3Hz,1H),8.52(d,J=3.8Hz,1H),8.33(d,J=8.7Hz,1H),7.30(d,J=7.9Hz,1H),4.98(s, 1H),4.17(s,2H),3.94–3.81(m,4H),3.60–3.49(m,2H),3.09–3.02(m,1H), 2.91(t,J=11.5Hz,2H),2.56(s,2H),2.09–1.84(m,3H),1.59-1.45(m,9H).

[0290] Example 7: (3S,4R)-4-((5-fluoro-4-(5-(2-hydroxypropan-2-yl)-6-isopropylimidazo[2,1-a]isoquinolin-8-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 7)

[0291] According to the synthesis method of compound 4-5, the raw material compound 4-1 is replaced by (isopropyl chloroformate) can obtain compound 7-1.

[0292] Step 1: Isopropyl 6-bromo-1-((2,4-dimethoxybenzyl)amino)-4-isopropylisoquinoline-3-carboxylate (Compound 7-2)

[0293] Compound 7-1 (1.0 g, 2.70 mmol) was dissolved in ultra-dry dimethyl sulfoxide (10 mL), potassium carbonate (745.71 mg, 5.40 mmol) and (2,4-dimethoxyphenyl)methylamine (902.18 mg, 5.40 mmol) were added, and the mixture was heated at 100°C under nitrogen for 16 hours. After the reaction was completed, the reaction system was added to water and extracted three times with ethyl acetate. The organic phases were combined and backwashed once with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated to dryness under reduced pressure, and purified by normal phase column chromatography (petroleum ether:ethyl acetate = 5:1-1:1) to obtain compound 7-2 (0.66 g, yield 49%).

[0294] LC-MS: m / z(ESI):501[M+H] + .

[0295] Step 2: 2-(6-bromo-1-((2,4-dimethoxybenzyl)amino)-4-isopropylisoquinolin-3-yl)propan-2-ol (Compound 7-3)

[0296] Compound 7-2 (150.0 mg, 299.16 μmol) was dissolved in anhydrous tetrahydrofuran (2 mL). Methylmagnesium bromide (3 M, 498.59 μL) was then added at 0°C. The mixture was allowed to react at room temperature for 2 hours. The reaction was then completed. The reaction system was added to a saturated aqueous ammonium chloride solution and extracted three times with ethyl acetate. The organic phases were combined and backwashed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain crude compound 7-3, which was used directly in the next reaction.

[0297] LC-MS: m / z(ESI):473[M+H] + .

[0298] Step 3: 2-(1-amino-6-bromo-4-isopropylisoquinolin-3-yl)propan-2-ol (Compound 7-4)

[0299] Crude compound 7-3 was dissolved in dichloromethane (2 mL), cooled to 0°C, and trifluoroacetic acid (0.5 mL) was slowly added. The reaction was then allowed to react at room temperature for 2 hours. The reaction was then concentrated to dryness under reduced pressure. The reaction solution was purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:9) to obtain compound 7-4 (25 mg, 26% yield over two steps).

[0300] LC-MS: m / z(ESI):323[M+H] + .

[0301] Step 4: 2-(8-bromo-6-isopropylimidazo[2,1-a]isoquinolin-5-yl)propan-2-ol (Compound 7-5)

[0302] Compound 7-4 (20 mg, 61.88 μmol) was dissolved in ethanol (2 mL), followed by the addition of sodium bicarbonate (10.40 mg, 123.75 μmol), chloroacetaldehyde (0.5 mL, 50% in H2O), and 4A molecular sieves (200 mg). The mixture was reacted at 80°C for 2 hours. The reaction was then concentrated to dryness under reduced pressure, and the reaction solution was purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:9) to obtain compound 7-5 (10 mg, 47% yield).

[0303] LC-MS: m / z(ESI):347[M+H] + .

[0304] Step 5: 2-(6-isopropyl-8-(trimethyltinyl)imidazo[2,1-a]isoquinolin-5-yl)propan-2-ol (Compound 7-6)

[0305] Under an argon atmosphere, compound 7-5 (10 mg, 28.80 μmol) was dissolved in dioxane (2 mL). Tetrakis(triphenylphosphine)palladium (3.3 mg, 2.88 μmol) and hexamethyltin (14.2 mg, 43.2 μmol) were then added, respectively. The reaction solution was stirred at 100°C for 1 hour. After the reaction was completed, the temperature was cooled to room temperature, filtered, and the filtrate was collected and concentrated under reduced pressure to obtain crude compound 7-6 (12.4 mg), which was used directly in the next reaction without purification.

[0306] Step 6: (3S,4R)-4-((5-fluoro-4-(5-(2-hydroxypropan-2-yl)-6-isopropylimidazo[2,1-a]isoquinolin-8-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-acetate (Compound 7-7)

[0307] Under an argon atmosphere, compound 7-6 (12.4 mg, 28.80 μmol) was dissolved in 1,4-dioxane (2 mL). Tetrakis(triphenylphosphine)palladium (3.3 mg, 2.88 μmol), cuprous chloride (2.8 mg, 28.8 μmol), and compound 2-7 (10.8 mg, 37.4 μmol) were then added. The reaction mixture was stirred at 100°C for 1 hour. After the reaction, the temperature was cooled to room temperature, filtered, and the filtrate was collected and concentrated under reduced pressure. The reaction mixture was purified by reverse phase column chromatography (eluent: water:acetonitrile = 1:9) to obtain compound 7-7 (6 mg, 40%).

[0308] LC-MS: m / z(ESI):522[M+H] + .

[0309] Step 7: (3S,4R)-4-((5-fluoro-4-(5-(2-hydroxypropan-2-yl)-6-isopropylimidazo[2,1-a]isoquinolin-8-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 7)

[0310] Compound 7-7 (14 mg, 26.87 μmol) was dissolved in a mixture of methanol (1 mL) and water (1 mL). Potassium carbonate (1.59 mg, 11.51 μmol) was then added and heated at 50°C for 1 hour to complete the reaction. The reaction solution was concentrated to dryness under reduced pressure and purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:9) to obtain compound 7 (1 mg, 8%).

[0311] LC-MS: m / z(ESI):480[M+H] + .

[0312] Example 8: (3S,4R)-4-((5-fluoro-4-(5-(2-hydroxypropyl-2-yl)-6-isopropyl-[1,2,4]triazolo[3,4-a]isoquinolin-8-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 8)

[0313] Step 1: 2-(6-bromo-1-chloro-4-isopropylisoquinolin-3-yl)propan-2-ol (Compound 8-1)

[0314] Compound 4-5 (100 mg, 238.83 μmol) was dissolved in tetrahydrofuran (2 mL) and purged with argon three times. Methylmagnesium bromide (3 M, 398.05 μL) was added at -78°C. After the addition was complete, the mixture was stirred for 5 minutes, then slowly warmed to room temperature and stirred for 30 minutes until the reaction was complete. The reaction mixture was quenched by adding saturated ammonium chloride solution and extracted three times with ethyl acetate. The organic layers were combined and concentrated to afford compound 8-1 (78 mg, yield: 95%).

[0315] m / z(ESI):342[M+H] + .

[0316] Step 2: 2-(6-Bromo-1-hydrazino-4-isopropylisoquinolin-3-yl)propan-2-ol (Compound 8-2)

[0317] Compound 8-1 (78 mg, 227.63 μmol), hydrazine hydrate (80%, 2 mL) and ethanol (5 mL) were added to a reaction flask and stirred at 80°C for 4 hours until the reaction was complete. The mixture was concentrated to obtain compound 8-2 (76 mg, yield: 99%).

[0318] m / z(ESI):338[M+H]+ .

[0319] Step 3: 2-(8-bromo-6-isopropyl-[1,2,4]triazolo[3,4-a]isoquinolin-5-yl)propan-2-ol (Compound 8-3)

[0320] Compound 8-2 (76 mg, 224.69 μmol) was added to trimethyl orthoformate (5 mL) and stirred at 90°C overnight until the reaction was complete. Compound 8-3 (30 mg, yield: 38%) was obtained by reverse phase chromatography (acetonitrile: water = 1:1).

[0321] m / z(ESI):348[M+H] + .

[0322] Step 4: (3S,4R)-4-((5-fluoro-4-(5-(2-hydroxypropan-2-yl)-6-isopropyl-[1,2,4]triazolo[3,4-a]isoquinolin-8-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-yl acetate (Compound 8-4)

[0323] Compound 8-3 (30 mg, 86.15 μmol), tetrakis(triphenylphosphine)palladium (9.95 mg, 8.61 μmol), and hexamethyltin (33.87 mg, 103.38 μmol) were added to a reaction flask, followed by 1,4-dioxane (3 mL). The mixture was purged with argon three times and allowed to react at 100°C for 1 hour, until the reaction was complete. Tetrakis(triphenylphosphine)palladium (9.95 mg, 8.61 μmol), compound 2-7 (27 mg, 94.76 μmol), and 1,4-dioxane (2 mL) were then added, purged with argon three times, and allowed to react at 100°C for 1 hour, until the reaction was complete. Compound 8-4 (10 mg, yield: 22%) was obtained by reverse phase chromatography (acetonitrile:water = 1:1) and purification.

[0324] m / z(ESI):523M+H] + .

[0325] Step 5: (3S,4R)-4-((5-fluoro-4-(5-(2-hydroxypropyl-2-yl)-6-isopropyl-[1,2,4]triazolo[3,4-a]isoquinolin-8-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 8)

[0326] Compound 8-4 (10 mg, 19.14 μmol) and hydrazine hydrate (80%, 1 mL) were added to ethanol (2 mL) and reacted at 60°C for 30 minutes. The reaction was complete. Reverse-phase chromatography (acetonitrile:water = 1:1) was used for purification to afford compound 8 (2.6 mg, yield: 28%).

[0327] m / z(ESI):481[M+H] + .

[0328] 1 H NMR (400MHz, DMSO-d6) δ9.77(s,1H),8.72(d,J=8.3Hz,1H),8.51(d,J=3.8Hz,1H),8.28(d,J=8.4Hz,1H),7.27(d,J=7.8Hz,1H),6.64(s,1H),5.32( s,1H),3.87(td,J=13.5,11.9,6.8Hz,4H),3.54(d,J=4.7Hz,2H),3.04(t, J=10.5Hz,1H),2.01(t,J=7.5Hz,3H),1.85(s,6H),1.58(t,J=6.9Hz,6H).

[0329] Example 9: (3S,4R)-4-((5-fluoro-4-(5-(2-hydroxypropan-2-yl)-6-isopropyl-[1,2,4]triazolo[5,1-a]isoquinolin-8-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 9)

[0330] Step 1: 1-amino-6-bromo-4-isopropylisoquinoline-3-carboxylic acid isopropyl ester (Compound 9-1)

[0331] Compound 7-2 (450 mg, 0.89 mmol) was dissolved in dichloromethane (10 mL), followed by the addition of trifluoroacetic acid (2 mL). The reaction mixture was allowed to react at room temperature for 3 hours. After completion of the reaction, the mixture was concentrated under reduced pressure and purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:1) to obtain compound 9-1 (280 mg, 90% yield).

[0332] m / z(ESI):351[M+H] + .

[0333] Step 2: (E)-6-bromo-1-(((dimethylamino)methylene)amino)-4-isopropylisoquinoline-3-carboxylic acid isopropyl ester (Compound 9-2)

[0334] Compound 9-1 (280 mg, 0.80 mmol) was dissolved in isopropanol (5 mL), followed by the addition of N,N-dimethylformamide dimethyl acetal (284 mg, 2.39 mmol). The reaction mixture was refluxed for 3 hours. After completion of the reaction, the crude product of compound 9-2 was obtained by direct concentration under reduced pressure and carried on to the next step without purification.

[0335] m / z(ESI):406[M+H] + .

[0336] Step 3: (E)-6-bromo-1-(N'-hydroxycarboxamido)-4-isopropylisoquinoline-3-carboxylic acid isopropyl ester (Compound 9-3)

[0337] Compound 9-2 (280 mg, 0.80 mmol) was dissolved in isopropanol (5 mL), followed by the addition of hydroxylamine hydrochloride (166 mg, 2.39 mmol). The reaction mixture was refluxed for 3 hours. After the reaction, the mixture was concentrated under reduced pressure to obtain the crude product of compound 9-3, which was directly carried out in the next step without purification.

[0338] m / z(ESI):394[M+H] + .

[0339] Step 4: 8-Bromo-6-isopropyl-[1,2,4]triazolo[5,1-a]isoquinoline-5-carboxylic acid isopropyl ester (Compound 9-4)

[0340] Compound 9-3 (150 mg, 0.38 mmol) was dissolved in tetrahydrofuran (5 mL) at 0°C, and trifluoroacetic anhydride (239 mg, 1.14 mmol) was slowly added. The reaction mixture was allowed to react at 0°C for 3 hours. After the reaction, the mixture was warmed to room temperature and purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:1) to obtain compound 9-4 (90 mg, 30% yield over three steps).

[0341] m / z(ESI):376[M+H] + .

[0342] Step 5: 2-(8-bromo-6-isopropyl-[1,2,4]triazolo[5,1-a]isoquinolin-5-yl)propan-2-ol (Compound 9-5)

[0343] Compound 9-4 (58 mg, 0.15 mmol) was dissolved in tetrahydrofuran (2 mL) at 0°C, and methylmagnesium bromide (0.5 mL, 1 M) was added. The reaction mixture was allowed to react at 0°C for 1 hour. After the reaction was completed, the mixture was warmed to room temperature and quenched with saturated ammonium chloride. The mixture was purified by reverse phase column chromatography (eluent: water:acetonitrile = 1:1) to obtain compound 9-5 (35 mg, 67%).

[0344] m / z(ESI):348[M+H] + .

[0345] Step 6: 2-(6-isopropyl-8-(trimethyltinyl)-[1,2,4]triazolo[5,1-a]isoquinolin-5-yl)propan-2-ol (Compound 9-6)

[0346] Under an argon atmosphere, compound 9-5 (20 mg, 57.43 μmol) was dissolved in dioxane (1.5 mL). Tetrakis(triphenylphosphine)palladium (6.6 mg, 5.74 μmol) and hexamethyltin (28 mg, 86.15 μmol) were then added, respectively. The reaction solution was stirred at 100°C for 1 hour. After the reaction was completed, the temperature was cooled to room temperature, filtered, and the filtrate was collected and concentrated under reduced pressure to obtain compound 9-6 (24 mg, crude product), which was used directly in the next reaction without purification.

[0347] m / z(ESI):434[M+H] + .

[0348] Step 6: (3S,4R)-4-((5-fluoro-4-(5-(2-hydroxypropan-2-yl)-6-isopropyl-[1,2,4]triazolo[5,1-a]isoquinolin-8-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-yl acetate (Compound 9-7)

[0349] Under an argon atmosphere, compound 9-6 (24 mg, 55.56 μmol) was dissolved in 1,4-dioxane (2 mL). Tetrakis(triphenylphosphine)palladium (6.41 mg, 5.55 μmol), cuprous chloride (5.5 mg, 55.56 μmol), and compound 2-7 (24 mg, 83 μmol) were then added. The reaction mixture was stirred at 100°C for 1 hour. After the reaction, the mixture was cooled to room temperature, filtered, and the filtrate was collected, concentrated under reduced pressure, and purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:1) to obtain compound 9-7 (11 mg, 38% yield).

[0350] m / z(ESI):523[M+H] + .

[0351] Step 7: (3S,4R)-4-((5-fluoro-4-(5-(2-hydroxypropan-2-yl)-6-isopropyl-[1,2,4]triazolo[5,1-a]isoquinolin-8-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 9)

[0352] Compound 9-7 (11 mg, 21.07 μmol) was dissolved in methanol / water (3:1, 2 mL), followed by the addition of potassium carbonate (8.71 mg, 63.15 μmol). The reaction was stirred at 50°C for 0.5 hours. Compound 9 (8 mg, 79% yield) was purified by reverse-phase column chromatography (eluent: water:acetonitrile = 2:1).

[0353] m / z(ESI):481[M+H] +

[0354] Example 10: (3S,4R)-4-((5-fluoro-4-(5-(1-hydroxycyclopentyl)-6-isopropyl-[1,2,4]triazolo[3,4-a]isoquinolin-8-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 10)

[0355] According to the synthesis method of compound 8, the methylmagnesium bromide in the first step is replaced by (dibromo-μ-1,4-butanediyldimagnesium) was synthesized to obtain compound 10.

[0356] m / z(ESI):507[M+H] +

[0357] Example 11: (3S,4R)-4-((5-fluoro-4-(4-(2-hydroxypropan-2-yl)-5-isopropyl-[1,2,4]triazolo[1,5-a]quinolin-7-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 11)

[0358] Step 1: 1-(2-amino-5-bromophenyl)-2-methylpropan-1-one (Compound 11-2)

[0359] Compound 11-1 (1 g, 5.08 mmol) was dissolved in tetrahydrofuran (10 mL). Isopropylmagnesium chloride (2 M, 8 mL) was added at 0°C and the mixture was stirred at room temperature for 12 hours to complete the reaction. Saturated aqueous ammonium chloride was added and the mixture was extracted three times with ethyl acetate. The organic phase was backwashed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. Purification by normal phase column chromatography (petroleum ether:ethyl acetate = 10:1 to 4:1) afforded the title compound 11-2 (1 g, 81% yield).

[0360] LC-MS: m / z(ESI):242[M+H] + .

[0361] Step 2: Ethyl 3-((4-bromo-2-isobutyrylphenyl)amino)-3-oxopropanoate (Compound 11-4)

[0362] Compound 11-2 (900.0 mg, 3.7 mmol) was dissolved in dichloromethane (15 mL). Ethyl 3-chloro-3-oxopropanoate (Compound 11-3, 840 mg, 5.6 mmol) and triethylamine (870 mg, 8.6 mmol) were added at 0°C and allowed to react for 1 hour. The reaction was completed. The mixture was quenched with water and extracted three times with ethyl acetate. The organic phase was backwashed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. The title compound 11-4 (1 g, 76% yield) was obtained by purification by normal phase column chromatography (petroleum ether:ethyl acetate = 10:1-4:1).

[0363] LC-MS: m / z(ESI):356[M+H] + .

[0364] Step 3: 6-bromo-4-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid ethyl ester (Compound 11-5)

[0365] Dissolve the crude compound 11-4 (670.0 mg, 1.9 mmol) in ethanol (7 mL), add sodium ethoxide (68 mg, 1 mmol), and react at 20°C for 1 hour. Add saturated ammonium chloride, extract three times with dichloromethane, and concentrate the organic phase to dryness under reduced pressure to obtain the crude title compound 11-5 (600 mg).

[0366] LC-MS: m / z(ESI):338[M+H] + .

[0367] Step 4: Ethyl 6-bromo-2-chloro-4-isopropylquinoline-3-carboxylate (Compound 11-6)

[0368] The crude compound 11-5 (600.0 mg) was dissolved in acetonitrile (6 mL), and phosphorus oxychloride (1.16 g, 7.6 mmol) was added. The reaction was allowed to proceed at 80°C for 8 hours. Saturated aqueous sodium bicarbonate solution was added, and the mixture was extracted three times with ethyl acetate. The organic phase was backwashed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain the crude compound 11-6 (620 mg).

[0369] LC-MS: m / z(ESI):356[M+H] + .

[0370] Step 5: 6-bromo-2-((2,4-dimethoxybenzyl)amino)-4-isopropylquinoline-3-carboxylic acid ethyl ester (Compound 11-7)

[0371] Compound 11-6 (100.0 mg, 0.28 mmol) and (2,4-dimethoxyphenyl)methanamine (94 mg, 0.56 mmol) were dissolved in dimethyl sulfoxide (2 mL), and potassium carbonate (75.0 mg, 0.55 mmol) was added. The reaction was heated at 80°C under nitrogen for 12 hours to complete the reaction. Saturated aqueous ammonium chloride was added, and the mixture was extracted three times with ethyl acetate. The organic phase was backwashed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. The title compound 11-7 (130 mg, 93% yield over three steps) was obtained by purification by normal phase column chromatography (petroleum ether:ethyl acetate = 10:1 to 1:1).

[0372] LC-MS: m / z(ESI):487[M+H] + .

[0373] Step 6: 2-amino-6-bromo-4-isopropylquinoline-3-carboxylic acid ethyl ester (Compound 11-8)

[0374] Compound 11-7 (130.0 mg, 0.27 mmol) was dissolved in trifluoroacetic acid (2 mL) and allowed to react at room temperature for 1 hour. The reaction mixture was then concentrated to dryness under reduced pressure, and saturated aqueous sodium bicarbonate solution was added. The mixture was extracted three times with ethyl acetate. The organic phase was backwashed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain the crude title compound 11-8 (100.0 mg).

[0375] LC-MS: m / z(ESI):337[M+H] + .

[0376] Step 7: Ethyl 6-bromo-2-(N'-hydroxycarbamimidoyl)-4-isopropylquinoline-3-carboxylate (Compound 11-9)

[0377] Dissolve the crude compound 11-8 (100.0 mg) in isopropanol (2 mL), add N,N-dimethylformamide dimethyl acetal (180 mg, 1.52 mmol), and heat to 80°C for 1 hour under nitrogen. Cool to 50°C, add hydroxylamine hydrochloride (121 mg, 1.77 mmol), and heat to 50°C for 1 hour to complete the reaction. Concentrate the reaction solution under reduced pressure to dryness to obtain the crude title compound 11-9 (350 mg).

[0378] LC-MS: m / z(ESI):380[M+H] + .

[0379] Step 8: 7-Bromo-5-isopropyl-[1,2,4]triazolo[1,5-a]quinoline-4-carboxylic acid ethyl ester (Compound 11-10)

[0380] The crude compound 11-9 (350.0 mg) was dissolved in tetrahydrofuran (2 mL), and trifluoroacetic anhydride (316 mg, 1.5 mmol) was added. The reaction was allowed to react at 25°C for 1 hour, and the reaction was completed. Saturated aqueous sodium bicarbonate solution was added, and the mixture was extracted three times with ethyl acetate. The organic phase was backwashed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. Purification by normal phase column chromatography (petroleum ether:ethyl acetate = 10:1-4:1) gave the title compound 11-10 (90 mg, 96% yield over three steps).

[0381] LC-MS: m / z(ESI):362[M+H] + .

[0382] Step 9: 2-(7-Bromo-5-isopropyl-[1,2,4]triazolo[1,5-a]quinolin-4-yl)propan-2-ol (Compound 11-11)

[0383] Compound 11-10 (100.0 mg, 0.28 mmol) was dissolved in tetrahydrofuran (2 mL). Methylmagnesium bromide (3 M, 0.5 ml) was added at 0°C and the mixture was allowed to react at room temperature for 1 hour. Saturated aqueous ammonium chloride was added, and the mixture was extracted three times with ethyl acetate. The organic phase was backwashed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. Purification by normal phase column chromatography (petroleum ether:ethyl acetate = 10:1 to 1:1) afforded the title compound 11-11 (30 mg, 31% yield).

[0384] LC-MS: m / z(ESI):348[M+H] + .

[0385] Step 10: 2-(5-isopropyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,2,4]triazolo[1,5-a]quinolin-4-yl)propan-2-ol (Compound 11-12)

[0386] Compound 11-11 (20 mg, 0.057 mmol) was dissolved in 1,4-dioxane (1 mL). [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (4.3 mg, 0.006 mmol), bipyralidoborane (21.5 mg, 0.086 mmol), and potassium acetate (16.9 mg, 0.17 mmol) were added. The mixture was heated at 80°C under a nitrogen atmosphere for 2 hours to complete the reaction. The reaction mixture was filtered and the filtrate was concentrated to dryness under reduced pressure to obtain the crude title compound 11-12 (22 mg).

[0387] LC-MS: m / z(ESI):396[M+H] + .

[0388] Step 11: (3S,4R)-4-((5-fluoro-4-(4-(2-hydroxypropan-2-yl)-5-isopropyl-[1,2,4]triazolo[1,5-a]quinolin-7-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-yl acetate (Compound 11-14)

[0389] Compound 11-12 (22 mg) was dissolved in a mixture of 1,4-dioxane (1 mL) and water (0.1 mL). Tetrakis(triphenylphosphine)palladium (6.7 mg, 0.006 mmol), compound 2-7 (32 mg, 0.111 mmol), and sodium carbonate (17.1 mg, 0.165 mmol) were then added. The mixture was heated at 90°C under a nitrogen atmosphere for 16 hours, completing the reaction. The reaction solution was concentrated to dryness under reduced pressure and purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:1) to afford the title compound 11-14 (20 mg, 67% yield over two steps).

[0390] LC-MS: m / z(ESI):523[M+H] + .

[0391] Step 12: (3S,4R)-4-((5-fluoro-4-(4-(2-hydroxypropan-2-yl)-5-isopropyl-[1,2,4]triazolo[1,5-a]quinolin-7-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 11)

[0392] Compound 11-14 (10.0 mg, 0.019 mmol) was dissolved in a mixture of methanol (1 mL) and water (1 mL). Potassium carbonate (14.0 mg, 0.1 mmol) was then added and heated at 50°C for 1 hour. The reaction was completed. The reaction solution was purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:1) to obtain compound 11 (3.0 mg, 33% yield).

[0393] LC-MS: m / z(ESI):481[M+H] + .

[0394] 1H NMR (400MHz, DMSO-d6) δ9.25(s,1H),8.60(d,J=8.7Hz,1H),8.57(s,1H),8.51(d,J=3.9Hz,1H),8.40(d,J=8.7Hz,1H),7.25(d,J=7.9Hz,1H),5.9 7(s,1H),4.95(d,J=5.3Hz,1H),3.90–3.83(m,4H),3.58–3.53(m,1H),3 .10–3.03(m,1H),2.01–1.96(m,3H),1.88(s,6H),1.60(t,J=7.2Hz,6H).

[0395] Example 12: (3S,4R)-4-((5-fluoro-4-(4-methyl-[1,2,3]triazolo[1,5-a]quinolin-7-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 12)

[0396] Step 1: (E)-N-(4-chlorophenyl)-2-methyl-3-phenylacrylamide (Compound 12-3)

[0397] Compound 12-1 (2 g, 15.67 mmol) was dissolved in acetonitrile (20 mL), followed by the addition of (E)-2-methyl-3-phenyl-2-propenoic acid (2.36 g, 14.55 mmol), N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (10.36 g, 36.99 mmol), and 2,4,6-trimethylpyridine (7.47 g, 61.66 mmol). The reaction mixture was stirred at room temperature for 30 minutes, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and purified by normal phase column chromatography (eluent: petroleum ether:ethyl acetate = 10:1) to obtain compound 12-3 (1.1 g, 26%).

[0398] m / z(ESI):272[M+H] +

[0399] Step 2: 6-chloro-3-methylquinolin-2(1H)-one (Compound 12-4)

[0400] Compound 12-3 (1.10 g, 4.06 mmol) was dissolved in tetrahydrofuran (10 mL) at 0°C, and aluminum chloride (3.24 g, 24.29 mmol) was added portionwise. The reaction mixture was heated to 120°C and reacted for 1 hour. After the reaction, the mixture was cooled to room temperature, quenched with ice water, and filtered directly. The filter residue was collected to obtain compound 12-4 (625 mg, 80%).

[0401] m / z(ESI):194[M+H] +

[0402] Step 3: 2,6-dichloro-3-methylquinoline (Compound 12-5)

[0403] Compound 12-4 (625 mg, 3.24 mmol) was dissolved in acetonitrile (10 mL), followed by the addition of phosphorus oxychloride (1.48 g, 9.68 mmol). The reaction mixture was heated to 80°C for 3 hours. After completion of the reaction, the mixture was cooled to room temperature and concentrated under reduced pressure to directly obtain compound 12-5 (525 mg, 76%), which was used directly in the next step without purification.

[0404] m / z(ESI):212[M+H] +

[0405] Step 4: (6-chloro-3-methylquinolin-2-yl)methanol (Compound 12-6)

[0406] Under an argon atmosphere, compound 12-5 (525 mg, 2.48 mmol) was dissolved in dioxane (5 mL). Tetrakis(triphenylphosphine)palladium (285 mg, 247.55 μmol) and (tributyltin)methanol (1.19 g, 3.71 mmol) were then added. The reaction mixture was stirred at 100°C for 12 hours. After the reaction was completed, the mixture was cooled to room temperature and purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:1) to obtain compound 12-6 (210 mg, 41%).

[0407] m / z(ESI):208[M+H] +

[0408] Step 5: 6-chloro-3-methylquinoline-2-carbaldehyde (Compound 12-7)

[0409] Compound 12-6 (200 mg, 975.61 μmol) was dissolved in dimethyl sulfoxide (3 mL), followed by the addition of 2-iodoacylbenzoic acid (404 mg, 1.44 mmol). The reaction mixture was then allowed to react at room temperature for 1 hour. Upon completion, saturated sodium bicarbonate solution was added to quench the reaction, followed by extraction with ethyl acetate, drying over anhydrous sodium sulfate, filtration, and purification by normal phase column chromatography (eluent: petroleum ether:ethyl acetate = 5:1) to afford compound 12-7 (135 mg, 67%).

[0410] m / z(ESI):206[M+H] +

[0411] Step 6: 7-chloro-4-methyl-[1,2,3]triazolo[1,5-a]quinoline (Compound 12-8)

[0412] Compound 12-7 (135 mg, 655.34 μmol) was dissolved in dimethyl sulfoxide (3 mL), followed by the addition of potassium phosphate (417 mg, 1.97 mmol) and p-toluenesulfonyl hydrazide (146 mg, 787.78 μmol). The reaction mixture was then incubated at 110°C for 6 hours. After the reaction, the mixture was cooled to room temperature and purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:1) to obtain compound 12-8 (110 mg, 77%).

[0413] m / z(ESI):218[M+H] +

[0414] Step 7: 4-Methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,2,3]triazolo[1,5-a]quinoline (Compound 12-9)

[0415] Under an argon atmosphere, compound 12-8 (25 mg, 115.21 μmol) was dissolved in dioxane (2 mL). [1,1'-bis(di-tert-butylphosphino)ferrocene]palladium dichloride (15 mg, 22.97 μmol), diboronic acid pinacol ester (146 mg, 574.31 μmol), and potassium acetate (34 mg, 344.58 μmol) were then added. The reaction mixture was stirred at 100°C for 1 hour. After the reaction, the temperature was cooled to room temperature, filtered, and the filtrate was collected and concentrated under reduced pressure to obtain crude compound 12-9 (30 mg, 84%), which was used directly in the next step without purification.

[0416] m / z(ESI):310[M+H] +

[0417] Step 8: (3S,4R)-4-((5-fluoro-4-(4-methyl-[1,2,3]triazolo[1,5-a]quinolin-7-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol acetate (Compound 12-10)

[0418] Under an argon atmosphere, compound 12-9 (30 mg, 97.03 μmol) was dissolved in dioxane / water (8:1, 1.9 ml). [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (7 mg, 9.70 μmol), compound 2-7 (42.16 mg, 145.55 μmol), and potassium carbonate (40.23 mg, 291.09 μmol) were then added. The reaction mixture was stirred at 100°C for 2 hours. After the reaction was completed, the mixture was cooled to room temperature and purified by reverse-phase column chromatography (eluent: water:acetonitrile = 1:1) to obtain compound 12-10 (20 mg, 47%).

[0419] m / z(ESI):437[M+H] +

[0420] Step 9: (3S,4R)-4-((5-fluoro-4-(4-methyl-[1,2,3]triazolo[1,5-a]quinolin-7-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol (Compound 12)

[0421] Compound 12-10 (20 mg, 45.83 μmol) was dissolved in methanol / water (3:1, 2 mL), followed by the addition of potassium carbonate (19.87 mg, 143.78 μmol). The reaction was stirred at 50°C for 0.5 hours. The product was purified by reverse-phase column chromatography (eluent: water:acetonitrile = 2:1) to obtain the desired product, compound 12 (16 mg, 88%).

[0422] m / z(ESI):395[M+H] +

[0423] 1 H NMR (400MHz, DMSO-d6) δ8.78(d,J=8.8Hz,1H),8.59(d,J=1.6Hz,1H),8.51(d,J=3 .7Hz,1H),8.43(s,1H),8.38(d,J=9.0Hz,1H),7.71(s,1H),7.26(d,J=7.7Hz,1H), 4.93(s,1H),3.91–3.75(m,3H),3.57–3.46(m,1H),3.42(s,1H),3.09(t,J=10.3Hz ,1H),2.59(d,J=1.3Hz,3H),2.03(d,J=13.1Hz,1H),1.51(qd,J=11.5,4.5Hz,1H).

[0424] Biological testing experiments

[0425] Test Example 1: Detection of CDK4 and CDK6 kinase inhibition effects

[0426] The CDK4 kinase assay method is as follows:

[0427] Perkin Elmer's Lance Ultra TR-FRET Kinase Assay reagents are used to measure CDK4 activity in vitro by measuring substrate phosphorylation during the kinase reaction.

[0428] The reaction buffer contained the following components: 50 mM HEPES (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 2 mM DTT, 0.01% Tween20;

[0429] Preparation of CDK4 kinase solution: Dilute human recombinant CDK4 / CycD1 protein (ProQinase, 0142-0143-1) with reaction buffer to a 3 nM kinase solution;

[0430] Preparation of substrate reaction solution: dilute 100 nM ULight-4E-BP1 kinase substrate (PerkinElmer, TRF0128) and 600 μM ATP with reaction buffer;

[0431] Preparation of detection buffer: Europium-anti-phospho-4E-BP1 antibody (PerkinElmer, TRF0216) and EDTA were diluted with 1× detection buffer (PerkinElmer, CR97-100) to 2 nM antibody solution and 20 mM EDTA solution, respectively.

[0432] Preparation of compound solutions of different concentrations: Using DMSO as the diluent, perform gradient dilution of the compound stock solution using the dose-response program of the pipette. The starting concentration of the test compound is 200 nM, and the solution is diluted 4-fold to 8 concentration points.

[0433] Using an Echo650 automated workstation, 100 nL of compound solutions of varying concentrations were added to a 384-well assay plate (Perkin Elmer, 6007299), followed by the addition of 5 μL of CDK4 kinase solution, mixed well, and incubated at room temperature for 5 minutes. 5 μL of substrate reaction solution was then added, and the reaction mixture was incubated at room temperature for 60 minutes. 10 μL of detection buffer equal in volume to the reaction was then added, mixed well, and allowed to stand at room temperature for 60 minutes. The reaction progress was detected using an Envision plate reader (Perkin Elmer) at wavelengths of 615 nm and 665 nm. The signal value (absorbance 665 nm / absorbance 615 nm) was positively correlated with the degree of phosphorylation of the substrate, thereby detecting the activity of CDK4 kinase. In this experiment, the group without CDK4 kinase protein was used as the 100% inhibition group, and the group with CDK4 kinase protein but no compound was used was used as the 0% inhibition group.

[0434] The inhibition percentage of CDK4 activity by the compound can be calculated using the following formula:

[0435] Inhibition percentage = 100-100*(signal value at specific concentration of test compound - signal value of 100% inhibition group) / (signal value of 0% inhibition group - signal value of 100% inhibition group).

[0436] The IC50 values ​​of the compounds were calculated from 8 concentration points using XLfit (ID Business Solutions Ltd., UK) software using the following formula:

[0437] Y=Bottom+(Top-Bottom) / (1+10^((logIC50-X)×slope factor))

[0438] Where Y is the inhibition percentage, X is the logarithm of the concentration of the test compound, Bottom is the minimum inhibition percentage, Top is the maximum inhibition percentage, and slope factor is the slope coefficient of the curve.

[0439] The CDK6 kinase assay method is as follows:

[0440] Perkin Elmer's Lance Ultra TR-FRET Kinase Assay reagents are used to measure CDK6 activity in vitro by measuring substrate phosphorylation during the kinase reaction.

[0441] The reaction buffer contained the following components: 50 mM HEPES (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 2 mM DTT, 0.01% Tween20;

[0442] Preparation of CDK6 kinase solution: Human recombinant CDK6 / CycD3 (Carna Biosciences, 04-107) protein was diluted with reaction buffer to a 2 nM kinase solution;

[0443] Preparation of substrate reaction solution: dilute 100 nM ULight-4E-BP1 kinase substrate (PerkinElmer, TRF0128) and 200 μM ATP with reaction buffer;

[0444] Preparation of detection buffer: Europium-anti-phospho-4E-BP1 antibody (PerkinElmer, TRF0216) and EDTA were diluted with 1× detection buffer (PerkinElmer, CR97-100) to 2 nM antibody solution and 20 mM EDTA solution, respectively.

[0445] Preparation of compound solutions of different concentrations: Using DMSO as the diluent, perform gradient dilution of the compound stock solution using the dose-response program of the pipette. The starting concentration of the test compound is 1 μM, and the solution is diluted 4-fold to 8 concentration points.

[0446] Use the Echo650 automated workstation to add 100nL of compound solutions of different concentrations to a 384-well detection plate (Perkin Elmer, 6007299), then add 5μL of CDK6 kinase solution, mix well and incubate at room temperature for 5 minutes. Subsequently, 5μL of substrate reaction solution was added, and the reaction mixture was incubated at room temperature for 60 minutes. Then, 10μL of detection buffer equal to the volume of the reaction was added, mixed well and allowed to stand at room temperature for 60 minutes, and the reaction progress was detected at wavelengths of 615nm and 665nm using an Envision plate reader (Perkin Elmer). The signal value (absorbance 665nm / absorbance 615nm) was positively correlated with the degree of phosphorylation of the substrate, thereby detecting the activity of CDK6 kinase. In this experiment, the group without CDK6 kinase protein was used as the 100% inhibition group, and the group with CDK6 kinase protein but no compound was added was used as the 0% inhibition group. The percentage of CDK6 activity inhibition by the compound can be calculated using the following formula:

[0447] Inhibition percentage = 100-100*(signal value at specific concentration of test compound - signal value of 100% inhibition group) / (signal value of 0% inhibition group - signal value of 100% inhibition group).

[0448] Compound IC 50 The value was calculated from 8 concentration points using XLfit (ID Business Solutions Ltd., UK) software using the following formula:

[0449] Y=Bottom+(Top-Bottom) / (1+10^((logIC 50 -X)×slope factor))

[0450] Where Y is the inhibition percentage, X is the logarithm of the concentration of the test compound, Bottom is the minimum inhibition percentage, Top is the maximum inhibition percentage, and slope factor is the slope coefficient of the curve.

[0451] Experimental results:

[0452] Test Example 2: MCF7 cell anti-proliferative activity experiment

[0453] Cell proliferation was quantified by measuring BrdU incorporation during DNA synthesis in replicating (cycling) cells using Sigma-Aldrich's Cell Proliferation ELISA, BrdU (11669915001) chemiluminescent detection reagent. ATCC-derived MCF7 cells were cultured to the logarithmic growth phase using the recommended medium. Cell pellets were obtained by trypsinization and centrifugation, and then counted. 40 μL of the system was plated at a density of 3,000 cells / well in 384-well plates (Corning, 3570) and cultured overnight. 40 nL of compound solutions of varying concentrations were added to 384-well assay plates (Perkin Elmer, 6007299) using an Echo 650 automated workstation (preparation method: using DMSO as the diluent, the compound stock solution was serially diluted using the dose-response program of the pipette instrument, with a starting concentration of 5 μM and 4-fold dilutions for 8 concentration points). After 24 hours of treatment, cell proliferation ELISA and BrdU (chemiluminescence) detection kits were used according to the manufacturer's instructions. Luminescence signal values ​​were measured using an Envision plate reader. The signal value is proportional to the amount of DNA synthesis in the cells, which is proportional to the cell proliferation rate, thereby detecting the proliferation activity of MCF7 cells. In this experiment, the group without cell addition was designated as the 100% inhibition group, and the group with cell addition but no compound addition was designated as the 0% inhibition group.

[0454] The inhibition percentage of the compound on MCF7 cell proliferation activity can be calculated using the following formula:

[0455] Inhibition percentage = 100-100*(signal value at specific concentration of test compound - signal value of 100% inhibition group) / (signal value of 0% inhibition group - signal value of 100% inhibition group).

[0456] Compound IC 50 The value was calculated from 8 concentration points using XLfit (ID Business Solutions Ltd., UK) software using the following formula:

[0457] Y=Bottom+(Top-Bottom) / (1+10^((logIC50-X)×slope factor))

[0458] Where Y is the inhibition percentage, X is the logarithm of the concentration of the test compound, Bottom is the minimum inhibition percentage, Top is the maximum inhibition percentage, and slope factor is the slope coefficient of the curve.

[0459] Test Example 3: T47D cell anti-proliferative activity experiment

[0460] Human breast cancer T47D cells used in this experiment were purchased from Kangyuan Bochuang (Cat. No. KC-0199). They were cultured in RPMI-1640 medium (Shanghai Yuanpei Biotechnology Co., Ltd., L240KJ) supplemented with 10% FBS (Longsa Biotechnology Co., Ltd., S211201T) and 8 μg / mL human insulin (Yisheng Biotechnology Co., Ltd., 40112ES60). Cell proliferation was quantified by detecting BrdU incorporation during DNA synthesis in replicating cells using a Sigma-Aldrich cell proliferation ELISA and BrdU chemiluminescence detection kit (Cat. No. 11669915001). T47D cells were digested with trypsin (Shanghai Yuanpei Biotechnology Co., Ltd., S310KJ) and centrifuged to obtain a cell pellet. After cell counting, 3000 cells were plated in 40 μL of the medium in a 384-well plate (Corning, 3570) and cultured overnight. Using an Echo 650 automated workstation, 40 nL of compound solution or DMSO at varying concentrations was added to a 384-well plate. Compound solution preparation method: Using DMSO as the diluent, a dose-response protocol was used to perform a serial dilution of the compound, starting at 3 μM and followed by three-fold dilutions over eight concentrations. The concentration range was 3 μM to 1.37 nM, with a final DMSO concentration of 0.1%. After incubation for 24 hours at 37°C in a 5% CO2 incubator, the cell proliferation assay was performed using a cell proliferation ELISA and BrdU detection kit according to the manufacturer's instructions. Luminescence signal was measured using an Envision microplate reader (PerkinElmer, Envision 2105). The signal is proportional to the amount of DNA synthesis in the cells, which in turn is proportional to the rate of cell proliferation, thereby measuring the proliferation activity of T47D cells.

[0461] In this experiment, the group in which cells were not added with the BrdU labeling reagent was designated as the 100% inhibition group, and the group in which cells were added but no compound was added was designated as the 0% inhibition group.

[0462] The inhibition percentage of the compound on T47D cell proliferation activity can be calculated using the following formula:

[0463] Inhibition percentage = 100*(signal value of 0% inhibition group - signal value at specific concentration of test compound) / (signal value of 0% inhibition group - signal value of 100% inhibition group).

[0464] XLfit (ID Business Solutions Ltd., UK) software was used to fit the inhibition curve and calculate the half-maximal inhibitory concentration (IC) using the following formula: 50 ): Y=Bottom+(Top-Bottom) / (1+10^((logIC50 -X)×slope factor))

[0465] Where Y is the inhibition percentage, X is the logarithm of the concentration of the test compound, Bottom is the minimum inhibition percentage, Top is the maximum inhibition percentage, and slope factor is the slope coefficient of the curve.

[0466] Experimental results:

Claims

1. A compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, in: X and Y are independently selected from N and CR 1 ; X 1 and X 2 independently selected from N, C and CH; Ring A is selected from 4-10 membered heterocyclic rings and 5-12 membered heteroaromatic rings, wherein the 4-10 membered heterocyclic rings and 5-12 membered heteroaromatic rings are optionally substituted by one or more R a replace; Ring B is selected from C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-12 membered heteroaryl, the C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-12 membered heteroaryl are optionally substituted by one or more R b replace; R 1 Selected from hydrogen, halogen, hydroxyl, C1-C 10 Alkyl, C2-C 10 Alkenyl and C2-C 10 Alkynyl, the hydroxyl, C1-C 10 Alkyl, C2-C 10 Alkenyl and C2-C 10 Alkynyl is optionally substituted with one or more R 1a replace; R 2 Selected from hydrogen, halogen, hydroxyl, amino, mercapto, cyano, C1-C 10 Alkyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl, the hydroxyl, amino, thiol, C1-C 10 Alkyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl are optionally substituted by one or more R 2a replace; R 3 、R 4 and R 7 independently selected from hydrogen, halogen, hydroxy, cyano, C1-C 10 Alkyl, C1-C 10 Alkoxy and C1-C4 haloalkyl; R 5 Selected from hydrogen, C2-C 10 Alkenyl, C2-C 10 Alkynyl and C1-C 10 Alkyl, the C2-C 10 Alkenyl, C2-C 10 Alkynyl and C1-C 10 The alkyl group is optionally substituted with one or more R 5a replace; R 6 Selected from hydrogen, halogen, hydroxy, cyano, C1-C 10 Alkyl, C2-C 10 Alkenyl and C2-C 10 Alkynyl, the hydroxyl, C1-C 10 Alkyl, C2-C 10 Alkenyl and C2-C 10 Alkynyl is optionally substituted with one or more R 6a replace; Every R 1a independently selected from halogen, hydroxy, cyano, amino and C1-C 10 alkyl; Every R 2a Independently selected from halogen, hydroxy, cyano, amino, C1-C8 alkyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl, the hydroxyl, amino, C1-C8 alkyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl are optionally substituted by one or more R c replace; Every R 5a 、R 6a independently selected from halogen, hydroxy, cyano and amino; Every R a independently selected from halogen, hydroxy, cyano, amino, C1-C4 alkyl, C1-C4 haloalkyl and C1-C4 alkoxy; Every R b independently selected from halogen, hydroxy, amino, cyano, -C(O)-R e 、-OC(O)-R f 、-S(O)2-R e 、C2-C 10 Alkenyl, C2-C 10 Alkynyl, C1-C 10 Alkyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl, the hydroxyl, amino, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C1-C 10 Alkyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl are optionally substituted by one or more R d replace; Every R c and R d independently selected from cyano, amino, halogen, hydroxy, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl, the amino, hydroxyl, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl are optionally substituted by one or more R g replace; Every R e 、R f and R g Independently selected from halogen, amino, C1-C4 alkyl, C1-C4 alkoxy and 4-10 membered heterocyclyl, wherein the amino, C1-C4 alkyl, C1-C4 alkoxy and 4-10 membered heterocyclyl are optionally substituted by amino, halogen, hydroxyl and C1-C4 alkyl; One or more hydrogen atoms of the compound are optionally deuterium atoms.

2. The compound of formula (I) according to claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: At least one of X and Y is CR 1 ; or X is N, Y is CR 1 ; or X and Y are both CR 1 .

3. The compound of formula (I) according to any one of claims 1 to 2, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: X 1 and X 2 At least one is C; or X 1 and X 2 All are C; or X 1 It is N, X 2 It is C; or X 1 It's C, X 2 It's N.

4. The compound of formula (I) according to any one of claims 1 to 3, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Ring A is selected from 4-7 membered heterocyclic rings and 5-10 membered heteroaromatic rings, wherein the 4-7 membered heterocyclic rings and 5-10 membered heteroaromatic rings are optionally substituted by one or more R a or Ring A is selected from 5-6 membered heterocyclic rings and 5-6 membered heteroaromatic rings, wherein the 5-6 membered heterocyclic rings and 5-6 membered heteroaromatic rings are optionally substituted by one or more R a or ring A is selected from pyrazole ring, imidazole ring, triazole ring, thiazole ring, oxazole ring, dihydrofuran ring and pyridine ring, wherein the pyrazole ring, imidazole ring, triazole ring, thiazole ring, oxazole ring, dihydrofuran ring and pyridine ring are optionally substituted by one or more R a or ring A is selected from a triazole ring, a thiazole ring, an oxazole ring, a dihydrofuran ring and a pyridine ring, wherein the triazole ring, the thiazole ring, the oxazole ring, the dihydrofuran ring and the pyridine ring are optionally substituted with one or more R a replace.

5. The compound of formula (I) according to any one of claims 1 to 4, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Every R a Independently selected from halogen, hydroxy, cyano, amino, C1-C4 alkyl.

6. The compound of formula (I) according to any one of claims 1 to 5, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Selected from or Selected from or Selected from 7. The compound of formula (I) according to any one of claims 1 to 6, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Ring B is selected from 4-10 membered heterocyclic groups and 5-12 membered heteroaryl groups, wherein the 4-10 membered heterocyclic groups and 5-12 membered heteroaryl groups are optionally substituted by one or more R b or Ring B is selected from 4-7 membered heterocyclic groups and 5-10 membered heteroaryl groups, wherein the 4-7 membered heterocyclic groups and 5-10 membered heteroaryl groups are optionally substituted by one or more R b or Ring B is selected from 5-6 membered heterocyclic groups and 5-6 membered heteroaryl groups, wherein the 5-6 membered heterocyclic groups and 5-6 membered heteroaryl groups are optionally substituted by one or more R b or ring B is selected from tetrahydropyranyl, piperidinyl and pyridinyl, wherein the tetrahydropyranyl, piperidinyl and pyridinyl are optionally substituted with one or more R b or ring B is selected from tetrahydropyranyl and pyridinyl, wherein the tetrahydropyranyl and pyridinyl are optionally substituted by one or more R b Substituted; or Ring B is selected from Or ring B is 8. The compound of formula (I) according to any one of claims 1 to 7, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Every R b independently selected from halogen, hydroxy, amino, cyano, -C(O)-R e 、-OC(O)-R f 、C2-C 10 Alkenyl, C2-C 10 Alkynyl, C1-C 10 Alkyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl, the hydroxyl, amino, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C1-C 10 Alkyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl are optionally substituted by one or more R d Replacement; or R b independently selected from halogen, hydroxy, -C(O)-R e 、-OC(O)-R f 、-S(O)2-R e 、C1-C 10 Alkyl and 4-10 membered heterocyclic group, the hydroxyl group, C1-C 10 The alkyl group and the 4-10 membered heterocyclic group are optionally substituted by one or more R d Replace; or each R b independently selected from halogen, hydroxy, -C(O)-R e 、-OC(O)-R f 、-S(O)2-R e , C1-C4 alkyl and 5-6 membered heterocyclic group, wherein the hydroxyl group, C1-C4 alkyl and 5-6 membered heterocyclic group are optionally substituted by one or more R d Replace; or each R b independently selected from halogen, hydroxy, -C(O)-R e 、-OC(O)-R f 、C1-C 10 Alkyl and 4-10 membered heterocyclic group, the hydroxyl group, C1-C 10 The alkyl group and the 4-10 membered heterocyclic group are optionally substituted by one or more R d Replace; or each R b independently selected from halogen, hydroxy, -C(O)-R e 、-OC(O)-R f , C1-C4 alkyl and 4-7 membered heterocyclic group, wherein the hydroxyl group, C1-C4 alkyl and 4-7 membered heterocyclic group are optionally substituted by one or more R d Replace; or each R b independently selected from fluoro, hydroxy, -C(O)-R e 、-OC(O)-R f 、-S(O)2-R e , methyl, piperidinyl and piperazinyl, wherein the hydroxyl, methyl, piperidinyl and piperazinyl groups are optionally substituted by one or more R d Replace; or each R b independently selected from fluoro, hydroxy, -C(O)-R e 、-OC(O)-R f , methyl, piperidinyl and piperazinyl, wherein the hydroxyl, methyl, piperidinyl and piperazinyl groups are optionally substituted by one or more R d Replace; or each R b Independently selected from fluorine, hydroxyl, Or each R b Independently selected from fluorine, hydroxyl, 9. The compound of formula (I) according to any one of claims 1 to 8, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Every R e independently selected from amino, C1-C4 alkyl, C1-C4 alkoxy and 4-7 membered heterocyclyl, wherein the amino, C1-C4 alkyl, C1-C4 alkoxy and 4-7 membered heterocyclyl are optionally substituted with amino, halogen, hydroxyl, C1-C4 alkyl; or each R e independently selected from amino, C1-C4 alkoxy and 4-7 membered heterocyclyl, wherein the amino, C1-C4 alkoxy and 4-7 membered heterocyclyl are optionally substituted by amino, halogen, hydroxyl, C1-C4 alkyl; or each R e independently selected from methyl, N(CH3)2, methoxy and morpholinyl; or each R e Independently selected from N(CH3)2, methoxy and morpholinyl.

10. The compound of formula (I) according to any one of claims 1 to 9, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Every R f are independently selected from 4-10 membered heterocyclic groups, wherein the 4-10 membered heterocyclic groups are optionally substituted by amino, halogen, hydroxyl, C1-C4 alkyl; or each R f Independently selected from morpholinyl, the morpholinyl is optionally substituted with amino, halogen, hydroxyl, C1-C4 alkyl; or each R f are independently selected from morpholinyl.

11. The compound of formula (I) according to any one of claims 1 to 10, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Every R c and R d independently selected from amino, halogen, hydroxy, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl, the amino, hydroxyl, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C3-C 12 Cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl and 5-10 membered heteroaryl are optionally substituted by one or more R g replace.

12. The compound of formula (I) according to any one of claims 1 to 11, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Every R d Independently selected from C1-C8 alkyl and 4-10 membered heterocyclic group, the C1-C8 alkyl and 4-10 membered heterocyclic group are optionally substituted by one or more R g Replace; or each R d independently selected from methyl and piperazinyl, said methyl and piperazinyl being optionally substituted by one or more R g replace.

13. The compound of formula (I) according to any one of claims 1 to 12, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Every R g Independently selected from C1-C4 alkyl and halogen, the C1-C4 alkyl is optionally substituted by amino, halogen, hydroxyl; or each R g Independently selected from C1-C4 alkyl, the C1-C4 alkyl is optionally substituted by amino, halogen, hydroxyl; or each R g Independently selected from C1-C4 alkyl and halogen; or each R g Independently selected from ethyl and fluoro; or R g For ethyl.

14. The compound of formula (I) according to any one of claims 1 to 13, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: R 1 is selected from hydrogen and halogen; or R 1 is selected from hydrogen and fluorine; or R 1 It's hydrogen.

15. The compound of formula (I) according to any one of claims 1 to 14, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: R 2 Selected from amino, C1-C 10 Alkyl, C3-C 12 Cycloalkyl and 4-10 membered heterocyclic groups, the amino, C1-C 10 Alkyl, C3-C 12 Cycloalkyl and 4-10 membered heterocyclic groups are optionally substituted by one or more R 2a Replacement; or R 2 is selected from amino, C1-C4 alkyl, C3-C6 cycloalkyl and 4-7 membered heterocyclic groups, wherein the amino, C1-C4 alkyl, C3-C6 cycloalkyl and 4-7 membered heterocyclic groups are optionally substituted by one or more R 2a Replacement; or R 2 Selected from C1-C 10 Alkyl and 4-10 membered heterocyclic group, the C1-C 10 The alkyl group and the 4-10 membered heterocyclic group are optionally substituted by one or more R 2a Replacement; or R 2 Selected from C1-C4 alkyl and 4-7 membered heterocyclic group, wherein the C1-C4 alkyl and 4-7 membered heterocyclic group are optionally substituted by one or more R 2a Replacement; or R 2 is selected from methyl, ethyl, isopropyl, cyclopentyl, amino and tetrahydropyrrolyl, wherein the methyl, ethyl, isopropyl, cyclopentyl, amino and tetrahydropyrrolyl are optionally substituted by one or more R 2a Replacement; or R 2 is selected from methyl, ethyl, isopropyl and tetrahydropyrrolyl, wherein the methyl, ethyl, isopropyl and tetrahydropyrrolyl are optionally substituted by one or more R 2a Replacement; or R 2 Selected from methyl, aminomethyl, dimethylamino, or R 2 Selected from methyl, aminomethyl, or R 2 Selected from aminomethyl, 16. The compound of formula (I) according to any one of claims 1 to 15, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Every R 2a Independently selected from hydroxyl, amino, C1-C8 alkyl and 4-10 membered heterocyclic groups, wherein the hydroxyl, amino, C1-C8 alkyl and 4-10 membered heterocyclic groups are optionally substituted by one or more R c Replace; or each R 2a independently selected from hydroxyl, amino, C1-C4 alkyl and 4-7 membered heterocyclic groups, wherein the hydroxyl, amino, C1-C4 alkyl and 4-7 membered heterocyclic groups are optionally substituted by one or more R c Replace; or each R 2a Independently selected from hydroxyl, amino and 4-10 membered heterocyclic groups, wherein the hydroxyl, amino and 4-10 membered heterocyclic groups are optionally substituted by one or more R c Replace; or each R 2a Independently selected from hydroxyl, amino and 4-7 membered heterocyclic groups, wherein the hydroxyl, amino and 4-7 membered heterocyclic groups are optionally substituted by one or more R c Replace; or each R 2a independently selected from hydroxy, amino, tetrahydropyrrolyl, piperidinyl, methyl and morpholinyl, wherein the hydroxy, amino, tetrahydropyrrolyl, piperidinyl, methyl and morpholinyl are optionally substituted by one or more R c Replace; or each R 2a independently selected from hydroxy, amino, tetrahydropyrrolyl and morpholinyl, wherein the hydroxy, amino, tetrahydropyrrolyl and morpholinyl are optionally substituted by one or more R c Replace; or each R 2a Independently selected from hydroxy, amino, methyl, The hydroxyl, amino, methyl, Optional one or more R c Replace; or each of R 2a Independently selected from hydroxyl, amino, The hydroxyl group, amino group, Optional one or more R c Replace; or each R 2a Independently selected from hydroxyl, methyl and The hydroxyl group, methyl group and Optional one or more R c replace.

17. The compound of formula (I) according to any one of claims 1 to 16, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Every R c independently selected from cyano, halogen, hydroxyl, C1-C8 alkyl, said C1-C8 alkyl being optionally substituted with 1 or more halogens; or each R c independently selected from hydroxy, C1-C8 alkyl, said C1-C8 alkyl being optionally substituted with 1 or more halogens; or each R c independently selected from cyano, fluoro, hydroxy, methyl and ethyl, said methyl and ethyl being optionally substituted with one or more fluoro groups; or each R c Independently selected from hydroxy, methyl and ethyl, said methyl and ethyl being optionally substituted with one or more fluorine groups.

18. The compound of formula (I) according to any one of claims 1 to 17, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: R 3 and R 4 independently selected from hydrogen, halogen, hydroxy, cyano, C1-C 10 Alkyl and C1-C4 haloalkyl; or R 3 and R 4 are independently selected from hydrogen and halogen; or R 3 and R 4 are independently selected from hydrogen, chlorine and fluorine; or R 3 is selected from chlorine and fluorine, R 4 is hydrogen; or R 3 is fluorine, R 4 For hydrogen.

19. The compound of formula (I) according to any one of claims 1 to 18, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: R 5 For hydrogen.

20. The compound of formula (I) according to any one of claims 1 to 19, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: R 6 Selected from hydrogen and C1-C 10 Alkyl, the C1-C 10 The alkyl group is optionally substituted with one or more R 6a Replacement; or R 6 Selected from C1-C 10 Alkyl, the C1-C 10 The alkyl group is optionally substituted with one or more R 6a Replacement; or R 6 Selected from C1-C4 alkyl, the C1-C4 alkyl is optionally substituted by one or more R 6a Replacement; or R 6 is selected from hydrogen and isopropyl; or R 6 It is isopropyl.

21. The compound of formula (I) according to any one of claims 1 to 20, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: R 7 For hydrogen.

22. The compound of formula (I) or its stereoisomer or its pharmaceutically acceptable salt according to claim 1, selected from the compound of formula (II) or its stereoisomer or its pharmaceutically acceptable salt, in, X 3 and X 4 One of them is CH and the other is N; X 1 and X 2 Independently selected from C and N; X, Y, R 2 、R 3 、R 4 、R 5 、R 6 and Ring B is as defined in any one of claims 1 to 21.

23. The compound of formula (I) or its stereoisomer or its pharmaceutically acceptable salt according to claim 1, selected from the compound of formula (III) or its stereoisomer or its pharmaceutically acceptable salt, in, X 5 Selected from N and CR 8 ; R 8 is selected from hydrogen, halogen, cyano, hydroxy, amino, C1-C4 alkyl, C1-C4 haloalkyl and C1-C4 alkoxy; R 1 、R 2 、R 3 、R 6 and Ring B is as defined in any one of claims 1 to 21.

24. The compound of formula (I) according to claim 1 or its stereoisomer or pharmaceutically acceptable salt thereof is selected from the following compounds or pharmaceutically acceptable salts thereof, 25. A pharmaceutical composition comprising the compound of formula (I) according to any one of claims 1 to 24, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

26. Use of the compound of formula (I) according to any one of claims 1 to 24, or its stereoisomer or pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 25, in the preparation of a medicament for preventing or treating a CDK-mediated disease.

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