Compound used as CDK2 / 4 protein kinase inhibitor and application thereof

Abstract

Compounds useful as CDK2 / 4 protein kinase inhibitors and uses thereof are provided. Specifically, provided are compounds of formula I, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, solvates, isotope compounds or prodrugs thereof, in which the definitions of each group and substituent group are as described in the specification, and which are useful as inhibitors of cyclin dependent kinases (CDKs), in particular as inhibitors of Cyclin dependent kinases (CDKs), as well as inhibitors of Cyclin dependent kinases (CDKs), as well as inhibitors of Cyclin dependent kinases (CDKs). The compound is used for preventing or treating cell proliferative diseases and symptoms including various cancers, and is particularly used for regulating and / or treating related diseases caused by overexpression and / or abnormal activity of cyclin-dependent kinase CDK2 / 4.

Description

Compounds used as CDK2 / 4 protein kinase inhibitors and their applications Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a class of novel compounds used as CDK2 / 4 protein kinase inhibitors, and their use in regulating CDK2 / 4 protein kinase activity or treating CDK2 / 4-related cell proliferative diseases and conditions, especially cancer. Background Art

[0002] Protein kinases regulate a variety of biological functions, including DNA replication, transcription, translation, cell cycle progression, energy metabolism, migration, and cell growth, making them ideal targets for treating proliferative diseases and conditions, including cancer. There is a need for novel compounds that can selectively inhibit protein kinase activity and act as effective therapeutic antiproliferative agents.

[0003] Cyclin-dependent kinases (CDKs) belong to the serine / threonine kinase family. They exert their physiological functions by binding to corresponding cyclins (cyclins) to form active dimeric complexes, promoting cell growth and proliferation. Over 20 CDKs have been discovered and can be divided into two major categories based on their primary functions: those that regulate the cell cycle and those that regulate transcription. CDKs 1-6 and 14-18 and their cyclin partners (e.g., Cyclins A, B, D1, D2, D3, E, and F) are involved in the regulation of cell cycle progression and are considered cell cycle regulators. CDKs 7-13 and 19-20 and their cyclin partners (e.g., Cyclins C, H, K, L1, L2, T1, and T2) are involved in the regulation of transcription and are considered transcriptional regulators. CDKs are therefore involved in cell cycle control, apoptosis, differentiation, and transcriptional regulation. CDK inhibitors have been shown to be therapeutically effective in a variety of diseases, including cancer.

[0004] CDK4 / 6, upon binding to Cyclin D, regulates the cell cycle from G1 to S phase. Aberrations in the Cyclin D-CDK4 / 6-Rb pathway have been reported to be associated with the progression of resistance to endocrine therapy. Currently, several CDK4 / 6 inhibitors, such as palbociclib, ribociclib, and abemaciclib, have been approved for use in combination with endocrine therapy for the treatment of hormone receptor (HR)-positive, human epidermal growth factor 2 (HER2)-negative advanced or metastatic breast cancer. However, hematologic toxicities such as neutropenia and / or gastrointestinal side effects are common during treatment with CDK4 / 6 inhibitors, leading to discontinuation or intermittent dosing, severely impacting efficacy and adherence. Currently, research data suggests that the activity of the cell cycle protein cyclin D3-CDK6 may be involved in these side effects. At the same time, clinically, the CDK4 / 6 inhibitor palbociclib was found to produce drug resistance when combined with endocrine therapy. Analysis of transcriptional profiling data from patient samples revealed that the resistance mechanism was highly correlated with the activation of the MYC oncogene and Cyclin E-CDK2 activity. Currently, CDK2 has been proposed as a therapeutic target for a variety of cancers, such as KRAS-driven lung cancer, MYC-amplified neuroblastoma, melanoma, acute myeloid leukemia, glioblastoma, prostate cancer, and breast cancer. CDK2's cyclin binding partners (i.e., Cyclin E1 and Cyclin E2) are amplified in a considerable proportion of human cancers, and this phenotype indicates that patients have poor overall survival. By inhibiting Cyclin E-CDK2 activity, it is hoped that the resistance problem of current clinical CDK4 / 6 inhibitors can be overcome.

[0005] In summary, given the toxic side effects of current CDK4 / 6 dual-target inhibitors and the drug resistance they produce when used in combination with endocrine therapy, developing a selective CDK2 / 4 kinase inhibitor may have better safety and efficacy, as well as broader clinical application prospects.

[0006] Summary of the Invention

[0007] The purpose of the present invention is to provide a novel compound having CDK2 / 4 protein kinase inhibitory activity and better pharmacodynamics and pharmacokinetic properties.

[0008] In the first aspect of the present invention, a compound is provided, wherein the compound is a compound of formula I, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, solvate, isotope compound or prodrug thereof,

[0009] in:

[0010] X is N or CR5;

[0011] X1 is N or CR5;

[0012] R1 is selected from the following groups: H, halogen (such as F, Cl, Br), cyano, C 1-6 Alkyl (such as methyl, ethyl), C 1-6 Haloalkyl (such as CF3), C 1-6 Alkoxy (such as methoxy), C 3-6 Cycloalkyl (e.g., cyclopropyl);

[0013] R2 is as shown in Formula Ia or Ib;

[0014] in,

[0015] W3, W4 and W5 are each independently selected from: CH, N, NH;

[0016] One of W2 and W6 is C, and the other is C or N;

[0017] R6 are each independently selected from the following group: halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Hydroxyalkyl, C 3-6 Cycloalkyl, -CO-C 1-6 Alkyl (such as ),-COO-C 1-6 Alkyl (such as ), 4 to 6 membered heterocyclic group, -C 1-4 Alkylene-C 3-6 Cycloalkyl, -C 1-4 wherein the alkyl, alkylene, cycloalkyl and heterocyclic groups are optionally substituted by one or more substituents selected from the group consisting of halogen, hydroxy, cyano, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 alkoxy;

[0018] The subscript p is 0, 1, or 2;

[0019] W1 and W7 are each independently CH or N;

[0020] Subscript o is 0 or 1;

[0021] R7 are each independently selected from the group consisting of halogen, C 1-6 Alkyl, C 1-6 alkyl halide;

[0022] Subscript q is 0, 1, 2, or 3;

[0023] R3 are each independently selected from the following group: H, C 1-6 Alkyl (such as methyl), C 3-6 Cycloalkyl (such as cyclopropyl), hydroxyl, amino, C 1-6 Alkoxy (such as methoxy), C 1-6 Haloalkyl (such as CF3), C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, 5-8 membered heterocyclyl, 5-8 membered heteroaryl; or, two R3 located on the same atom together form an oxo group (=O);

[0024] R4 is selected from the following group: H, C 1-6 Alkyl (such as methyl, ethyl, isopropyl), -CO-C 1-6 Alkoxy (ie -CO-OC 1-6 Alkyl groups, such as ),-CO-C 1-6 Alkyl (such as ), C 3-8 Cycloalkyl (eg: ), a 4-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S (e.g.: ); wherein the alkyl, aryl, cycloalkyl and heterocycloalkyl groups are optionally substituted by one or more groups selected from the group consisting of halogen (such as F, Cl, or Br), cyano, hydroxyl, C 1-6 Alkyl, -CO-C 1-6 Alkyl, C 1-6 Alkoxy (such as -O-methyl), amino (such as NH2), -COOH, -CONH2, -CF2CF3, optionally C 1-6 Alkyl, -CH(F)2, -COO-C 1-6 Alkyl (such as -COO-C(CH3)3) or -S(O)2-C 6-10 aryl-substituted 5-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S;

[0025] R5 is selected from the following groups: H, halogen (such as F, Cl, Br), cyano, C 1-6 Alkyl (such as methyl, ethyl), C 1-6 Haloalkyl (such as CF3), -CO-C 1-6 Alkoxy (ie -CO-OC 1-6 Alkyl groups, such as )(Preferably, R5 is H);

[0026] The subscript m is selected from the following group: 0, 1, 2, 3, 4;

[0027] The subscript n is 0, 1, 2, 3 or 4 (preferably, 0, 1 or 2, more preferably, 0 or 1, and most preferably, 1).

[0028] In another preferred embodiment, the compound is a compound used as a CDK2 / 4 kinase inhibitor.

[0029] In another preferred embodiment,

[0030] X is N or CR5; and / or,

[0031] X1 is N or CR5; and / or,

[0032] R1 is selected from the following groups: H, halogen (such as F, Cl, Br), C 1-6 Alkyl (such as methyl), C 1-6 Haloalkyl (such as CF3), C 3-6 Cycloalkyl (such as cyclopropyl); and / or,

[0033] R2 is represented by Formula Ia or Ib; and / or,

[0034] in,

[0035] W3, W4 and W5 are each independently selected from: CH, N, NH; and / or,

[0036] One of W2 and W6 is C, and the other is C or N; and / or,

[0037] R6 are each independently selected from the following group: halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Hydroxyalkyl, C 3-6 Cycloalkyl, 4- to 6-membered heterocyclic group, -C 1-4 Alkylene-C 3-6 Cycloalkyl, -C 1-4 wherein the alkyl, alkylene, cycloalkyl and heterocyclic groups are optionally substituted by one or more substituents selected from the group consisting of halogen, hydroxy, cyano, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy; and / or,

[0038] subscript p is 0, 1, or 2; and / or,

[0039] W1 and W7 are each independently CH or N; and / or,

[0040] Subscript o is 0 or 1; and / or,

[0041] R7 are each independently selected from the group consisting of halogen, C 1-6 Alkyl, C 1-6 haloalkyl; and / or,

[0042] subscript q is 0, 1, 2, or 3; and / or,

[0043] R3 are each independently selected from the following group: H, C 1-6 Alkyl (such as methyl), C 3-6 Cycloalkyl (such as cyclopropyl), hydroxyl, C 1-6 Alkoxy (such as methoxy); and / or,

[0044] R4 is selected from the following group: H, C 1-6 Alkyl (such as methyl, ethyl, isopropyl), -CO-C 1-6 Alkoxy (ie -CO-OC 1-6 Alkyl groups, such as ),-CO-C 1-6 Alkyl (such as ); and / or,

[0045] R5 is selected from the following group: H, C 1-6 Alkyl (such as methyl, ethyl), -CO-C 1-6 Alkoxy (i.e. -CO-OC 1-6 Alkyl groups, such as )(Preferably, R5 is H); and / or,

[0046] The subscript m is selected from the group consisting of 0, 1, 2, 3, 4; and / or,

[0047] The subscript n is 0, 1, 2, 3 or 4 (preferably, 0, 1 or 2, more preferably, 0 or 1, and most preferably, 1).

[0048] In another preferred embodiment, R2 is as shown in Formula Ia.

[0049] In another preferred embodiment,

[0050] In formula (Ia),

[0051] W1 is CH, W2 is C, W3 is NH, W4 is CH, W5 is N, W6 is C and W7 is CH; or,

[0052] W1 is CH, W2 is C, W3 is CH, W4 is CH, W5 is N, W6 is N and W7 is CH; or,

[0053] W1 is CH, W2 is C, W3 is N, W4 is CH, W5 is N, W6 is N and W7 is CH; or,

[0054] W1 is CH, W2 is N, W3 is CH, W4 is CH, W5 is N, W6 is C and W7 is N; or,

[0055] W1 is CH, W2 is CH, W3 is N, W4 is CH, W5 is CH, W6 is N and W7 is CH; or,

[0056] W1 is CH, W2 is CH, W3 is N, W4 is N, W5 is CH, W6 is N and W7 is N; or,

[0057] W1 is CH, W2 is C, W3 is NH, W4 is CH, W5 is N, W6 is C and W7 is N; or,

[0058] W1 is CH, W2 is C, W3 is N, W4 is CH, W5 is N, W6 is N and W7 is N; or,

[0059] W1 is CH, W2 is C, W3 is NH, W4 is CH, W5 is CH, W6 is C and W7 is CH; or,

[0060] W1 is CH, W2 is C, W3 is NH, W4 is N, W5 is CH, W6 is C and W7 is CH; or,

[0061] W1 is CH, W2 is C, W3 is NH, W4 is CH, W5 is N, W6 is C and W7 is CH; or,

[0062] W1 is CH, W2 is C, W3 is CH, W4 is N, W5 is NH, W6 is C and W7 is CH; or,

[0063] W1 is CH, W2 is C, W3 is N, W4 is N, W5 is NH, W6 is C and W7 is CH; or,

[0064] W1 is CH, W2 is C, W3 is CH, W4 is NH, W5 is N, W6 is C and W7 is CH; or,

[0065] W1 is N, W2 is C, W3 is CH, W4 is NH, W5 is N, W6 is N and W7 is CH; or,

[0066] W1 is CH, W2 is C, W3 is CH, W4 is CH, W5 is N, W6 is N and W7 is N;

[0067] In formula (Ib),

[0068] W2 is C, W3 is CH, W4 is CH, W5 is N, W6 is N, and o is 0 or 1.

[0069] In another preferred embodiment, in formula (Ia), W1 is CH, W2 is C, W3 is CH, W4 is CH, W5 is N, W6 is N and W7 is N.

[0070] In another preferred embodiment, in formula (Ia), W1 is CH, W2 is C, W3 is NH, W4 is CH, W5 is N, W6 is C and W7 is CH.

[0071] In another preferred embodiment, in formula (Ia), W1 is CH, W2 is C, W3 is CH, W4 is NH, W5 is N, W6 is C and W7 is CH.

[0072] In another preferred embodiment, R6 are each independently selected from the following groups: halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Hydroxyalkyl, C 3-6 wherein the alkyl, alkylene, cycloalkyl and heterocyclic groups are optionally substituted by one or more substituents selected from the group consisting of halogen, hydroxy, cyano, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy group.

[0073] In another preferred embodiment, R6 are each independently selected from the following groups: halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Hydroxyalkyl, C 3-6 Cycloalkyl, 4 to 6 membered heterocycloalkyl, wherein the 4 to 6 membered heterocycloalkyl is selected from: pyrrolidinyl (such as ), piperidinyl (such as ).

[0074] In another preferred embodiment, in R6, the alkyl, alkylene, cycloalkyl and heterocyclic groups are unsubstituted.

[0075] In another preferred embodiment, Select from the following groups:

[0076] In another preferred embodiment, Select from the following groups:

[0077] In another preferred embodiment, Select from the following groups:

[0078] In another preferred embodiment, Select from the following groups:

[0079] In another preferred embodiment, X is N.

[0080] In another preferred embodiment, X1 is N.

[0081] In another preferred embodiment, the subscript n is 0 or 1.

[0082] In another preferred embodiment, R1 is selected from the following group: H, F, Cl, methyl, Br, CF3.

[0083] In another preferred embodiment, R3 are independently selected from the following groups: H, C 1-6 Alkyl, C 3-6 Cycloalkyl, hydroxyl, C 1-6 Alkoxy.

[0084] In another preferred embodiment, R3 is independently selected from the following group: H, methyl, cyclopropyl, hydroxy, methoxy, and oxo.

[0085] In another preferred embodiment, R3 is selected from the following group: H, methyl, cyclopropyl, hydroxyl, methoxy.

[0086] In another preferred embodiment, R4 is selected from the following group: H, C 1-6 Alkyl (such as methyl, ethyl, isopropyl), -CO-C 1-6 Alkoxy (i.e. -CO-OC 1-6 Alkyl groups, such as ),-CO-C 1-6 Alkyl (such as ), C 3-8 Cycloalkyl (eg: ), a 4-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S (e.g.: ); wherein the alkyl, cycloalkyl and heterocycloalkyl groups are optionally substituted by one or more groups selected from the group consisting of halogen (such as F, Cl, or Br), and hydroxyl.

[0087] In another preferred embodiment, R4 is selected from the following group: H, methyl, ethyl,

[0088] In another preferred embodiment, R4 is selected from the following group: H, methyl, ethyl,

[0089] In another preferred embodiment, the compound is as shown in Formula II;

[0090] Among them, R 31 The definition of is the same as R3, m1 is 0, 1, 2 or 3; R1, R2, R3, R4, X and X1 are the same as those in formula (I).

[0091] In another preferred embodiment, the compound is as shown in Formula II-1 or II-2;

[0092] Among them, R 31 The definition of is the same as R3, m1 is 0, 1, 2 or 3; R1, R2, R3, R4, X and X1 are the same as those in formula (I).

[0093] In another preferred embodiment, in Formula II, Formula II-1 or Formula II-2, R 31 H or C 1-6 Alkyl (such as methyl).

[0094] In another preferred embodiment, in Formula II, Formula II-1 or Formula II-2, R3 is independently selected from the following groups: H, C 1-6 Alkyl (such as methyl), C 3-6 Cycloalkyl (such as cyclopropyl), hydroxyl, C 1-6 Alkoxy (such as methoxy); or, two R3 located on the same atom together form an oxo group (=O).

[0095] In another preferred embodiment, the isotope compound is a deuterated compound.

[0096] In another preferred embodiment, the deuterated compound refers to a compound in which one hydrogen atom (H) or multiple hydrogen atoms (H) in the compound (preferably, in R4 and / or R2; more preferably, in R4 and / or R6) is replaced by a deuterium atom (D).

[0097] In another preferred embodiment, in the deuterated compound, R4 is deuterated C 1-6 Alkyl (such as -CD3).

[0098] In another preferred embodiment, in the deuterated compound, R6 is

[0099] In another preferred embodiment, X, X1, R1, R2, R3, R4, R5, subscript m and subscript n are each independently a corresponding group in the specific compounds shown in Table A or the Preparation Examples.

[0100] In another preferred embodiment, the compound is selected from Table A:

[0101] Table A

[0102] In another preferred embodiment, the inorganic acid salt is selected from the group consisting of hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, nitrate, phosphate, and acid phosphate.

[0103] In another preferred embodiment, the organic acid salt is selected from the group consisting of formate, acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, salicylate, picrate, glutamate, ascorbate, camphorate, and camphorsulfonate.

[0104] In the second aspect of the present invention, a pharmaceutical composition is provided, which contains a preventive and / or therapeutically effective amount of the compound as described in the first aspect and a pharmaceutically acceptable carrier.

[0105] In the third aspect of the present invention, there is provided a use of the compound according to the first aspect for preparing one or more drugs selected from the group consisting of:

[0106] (a) A drug used as a CDK2 / 4 protein kinase inhibitor;

[0107] (b) drugs for regulating CDK2 / 4 protein kinase activity;

[0108] (c) Drugs for preventing or treating CDK2 / 4-related diseases.

[0109] In another preferred embodiment, the CDK2 / 4-related disease is selected from the group consisting of inflammation, cancer, cardiovascular disease, infection, immune disease, and metabolic disease.

[0110] In another preferred embodiment, the cancer is selected from the group consisting of lung cancer, breast cancer, prostate cancer, colorectal cancer, liver cancer, pancreatic cancer, ovarian cancer, leukemia, neuroblastoma, gastric cancer, kidney cancer, esophageal cancer, uterine cancer, and liposarcoma.

[0111] In the fourth aspect of the present invention, a method for preventing or treating a CDK2 / 4-related disease is provided, comprising administering to a subject in need thereof a compound as described in the first aspect, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, solvate, isotope compound or prodrug thereof, or a pharmaceutical composition containing the compound, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, solvate, isotope compound or prodrug thereof.

[0112] In another preferred embodiment, the CDK2 / 4 related disease is as defined above.

[0113] In the fifth aspect of the present invention, a method for inhibiting CDK2 / 4 protein kinase is provided, comprising: contacting CDK2 / 4 protein kinase with the compound according to the first aspect, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, solvate, isotope compound or prodrug thereof, thereby inhibiting CDK2 / 4 protein kinase.

[0114] In another preferred embodiment, the method is non-therapeutic in vitro.

[0115] In a sixth aspect of the present invention, a method for regulating the activity of CDK2 / 4 protein kinase is provided, comprising: contacting CDK2 / 4 protein kinase with the compound according to the first aspect, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, solvate, isotope compound or prodrug thereof, thereby regulating the activity of CDK2 / 4 protein kinase.

[0116] In another preferred embodiment, the method is non-therapeutic in vitro.

[0117] In a seventh aspect of the present invention, a method for inhibiting cell proliferation is provided, comprising: contacting cells with the compound according to the first aspect, thereby inhibiting cell proliferation.

[0118] In another preferred embodiment, the cells are cancer cells or tumor cells.

[0119] In another preferred embodiment, the cells are selected from the group consisting of breast cancer cells (such as MCF-7), ovarian cancer cells (such as A2780), or a combination thereof.

[0120] In another preferred embodiment, the method is non-therapeutic in vitro.

[0121] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. DETAILED DESCRIPTION

[0122] After extensive and in-depth research, the inventors unexpectedly discovered a class of compounds with novel structures that exhibit promising CDK2 / 4 protein kinase inhibitory activity. These compounds exhibit excellent inhibitory activity against CDK2 / 4 protein kinases and possess enhanced pharmacodynamic and pharmacokinetic properties. Based on this discovery, the inventors completed the present invention.

[0123] the term

[0124] Unless otherwise stated, the following terms used in this application (including the specification and claims) have the definitions given below.

[0125] When a substituent is described by a conventional chemical formula written from left to right, the substituent also includes chemically equivalent substituents that would result if the formula were written from right to left. For example, -CH2O- is equivalent to -OCH2-.

[0126] As used herein, a group consisting of The bonds represented represent the point at which the group is attached to the rest of the compound or molecule.

[0127] "Alkyl", alone or as part of another group, refers to a monovalent straight or branched chain saturated hydrocarbon radical containing 1 to 12 carbon atoms (i.e., C 1-12 Alkyl). Alkyl is preferably C 1-6 Alkyl (i.e., an alkyl group containing 1, 2, 3, 4, 5, or 6 carbon atoms), more preferably C 1-4 Alkyl (i.e., an alkyl group containing 1, 2, 3, or 4 carbon atoms). Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, isobutyl, sec-butyl, tert-butyl, pentyl, n-hexyl, octyl, dodecyl, and the like. Unless otherwise indicated, in this application, alkyl is also intended to include substituted alkyl, i.e., one or more positions in the alkyl are substituted, especially 1-4 substituents, which may be substituted at any position. Unless otherwise indicated, in this application, "substituted alkyl" includes, for example, haloalkyl, hydroxyalkyl, and the like. As used herein, "haloalkyl" refers to an alkyl group as defined herein in which one or more hydrogen atoms in the group are replaced by the same or different halogen atoms. Haloalkyl is preferably C 1-6 Haloalkyl, more preferably C 1-4 Haloalkyl. Examples of haloalkyl include -CH2Cl, -CH2CF3, -CH2CCl3, perfluoroalkyl (e.g., -CF3), etc. As used herein, "hydroxyalkyl" refers to an alkyl group as defined herein in which one or more hydrogen atoms are replaced by hydroxyl groups. Hydroxyalkyl is preferably C 1-6 Hydroxyalkyl, more preferably C 1-4 Examples of hydroxyalkyl groups include -CH2OH, -C(CH3)2OH, and the like.

[0128] "Alkylene" refers to a divalent radical of an alkyl group as defined herein, preferably an alkylene group having 1 to 4 carbon atoms, i.e., C 1-4 Alkylene. Examples of alkylene include, for example, -CH2-, -CH2CH2-, and -CH2CH2CH2-.

[0129] "Alkoxy", alone or as part of another group, refers to an alkyl group having an oxygen group attached thereto, which has the structure alkylO-, wherein alkyl has the meaning as described above. Preferably, alkoxy is C 1-6 Alkoxy (ie -OC 1-6 alkyl), more preferably, C 1-4 Alkoxy (ie -OC 1-4 Alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, tert-butoxy, and the like. "Haloalkoxy" refers to a group of the formula -OR, where R is a haloalkyl group as defined herein. Examples of haloalkoxy groups include, but are not limited to, trifluoromethoxy, difluoromethoxy, 2,2,2-trifluoroethoxy, and the like.

[0130] "Thioalkyl" refers to an alkyl group as defined above where a carbon is replaced by S, S(O) or S(O)2, for example -SC 1-6 Alkyl, -S(O)C 1-6 Alkyl or -S(O)2C 1-6 alkyl.

[0131] "Alkenyl", alone or as part of another group, refers to an aliphatic group containing at least one double bond, typically having 2 to 20 carbon atoms (i.e., C 2-20 Alkenyl). Alkenyl is preferably C 2-6 Alkenyl (i.e., alkenyl containing 2, 3, 4, 5, or 6 carbon atoms). Alkenyl includes, but is not limited to, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, etc. Unless otherwise defined, in the present invention, alkenyl also includes substituted alkenyl.

[0132] "Alkenylene" refers to an alkenyl group as defined above with two points of attachment, for example, "vinylene" refers to the group -CH=CH-. Alkenylene is preferably C 2-6 Alkenylene (ie, alkenylene containing 2, 3, 4, 5, or 6 carbon atoms). Unless otherwise defined, alkenylene may be in unsubstituted or substituted form with one or more substituents.

[0133] "Alkynyl", alone or as part of another group, refers to a straight or branched hydrocarbon chain containing more than 2 carbon atoms and characterized by one or more triple bonds, typically having 2 to 20 carbon atoms (i.e., C 2-20 Alkynyl). Alkynyl is preferably C 2-6 Alkynyl (i.e., an alkynyl having 2, 3, 4, 5, or 6 carbon atoms). Alkynyl includes, but is not limited to, ethynyl, propargyl, and 3-hexynyl. One of the triple bond carbons may optionally be the point of attachment for an alkynyl substituent. In the present invention, unless otherwise defined, alkynyl also includes substituted alkynyl.

[0134] "Alkynylidene" refers to an alkynyl group as defined above with two points of attachment. For example, "ethynylene" refers to the group: -C≡C-. Alkynylidene is preferably C 2-6 Alkyneylene (ie, an alkynylene group containing 2, 3, 4, 5, or 6 carbon atoms). Unless otherwise defined, an alkynylene group may be unsubstituted or substituted with one or more substituents.

[0135] "Aliphatic group" refers to a straight-chain, branched-chain or cyclic hydrocarbon group, including saturated and unsaturated groups such as alkyl, alkenyl and alkynyl groups.

[0136] "Aromatic ring system" or "aromatic ring" refers to a monocyclic, bicyclic or polycyclic hydrocarbon ring system, wherein at least one ring is aromatic. Preferably, the "aromatic ring system" or "aromatic ring" has 6-12 ring atoms, i.e., C 6-12Aromatic ring. Examples of the aromatic ring include a benzene ring, a naphthalene ring, an anthracene ring, and the like.

[0137] "Aryl", alone or as part of another group, refers to a monovalent radical of an aromatic ring system (aromatic ring). Representative aryl groups include fully aromatic ring systems such as phenyl, naphthyl, and anthracenyl; and ring systems in which an aromatic carbocyclic ring is fused to one or more non-aromatic carbocyclic rings, such as indanyl, phthalimide, naphthylimide, or tetrahydronaphthyl, etc. In the present invention, aryl is preferably C 6-12 Aryl. In the present invention, unless otherwise defined, aryl also includes substituted aryl.

[0138] "Arylalkyl" or "aralkyl" refers to an alkyl moiety in which an alkyl hydrogen atom is replaced by an aryl group. Aralkyl groups include groups in which one or more hydrogen atoms in an alkyl group are replaced by an aryl group, wherein aryl and alkyl are as defined above. Examples of "arylalkyl" or "aralkyl" groups include benzyl, 2-phenylethyl, 3-phenylpropyl, 9-fluorenyl, diphenylmethyl, and triphenylmethyl.

[0139] "Aryloxy" means an -O-(aryl) group in which the aryl portion is as defined above.

[0140] "Heteroalkyl" refers to an alkyl group in which a carbon atom is replaced by an alkyl group having one or more backbone atoms selected from atoms other than carbon, for example, oxygen, nitrogen, sulfur, phosphorus, or a combination thereof. Numerical ranges may be given, for example, C 1-6 Heteroalkyl refers to the number of carbons in the chain, which includes 1 to 6 carbon atoms. For example, a -CH2OCH2CH3 group is referred to as a "C3" heteroalkyl. Attachment to the rest of the molecule can be through a heteroatom or carbon in the heteroalkyl chain. "Heteroalkylene" refers to a divalent alkyl group in which a carbon atom in the alkylene group is replaced by one or more backbone chain atoms selected from atoms other than carbon, such as oxygen, nitrogen, sulfur, phosphorus, or a combination thereof. Unless otherwise defined, "heteroalkyl" and "heteroalkylene" include substituted and unsubstituted forms.

[0141] "Carbocyclic ring system" or "carbocycle" refers to a monocyclic, bicyclic or polycyclic hydrocarbon ring system in which each ring is fully saturated or contains one or more unsaturated units, but in which none of the rings is aromatic. Preferably, a "carbocyclic ring system" or "carbocycle" has 6 to 12 ring atoms, i.e., C 6-12 Carbocyclic. "Carbocyclyl" refers to a monovalent group of a carbocyclic ring system or carbocyclic ring as defined above. Preferably, the carbocyclyl has 6-12 ring atoms, i.e., C 6-12 Examples of carbocyclic groups include cycloalkyl groups (e.g., cyclopentyl, cyclobutyl, cyclopropyl, cyclohexyl, etc.) and cycloalkenyl groups (e.g., cyclopentenyl, cyclohexenyl, cyclopentadienyl, etc.).

[0142] "Cycloalkyl" refers to a monovalent saturated carbocyclic group consisting of a mono- or bicyclic ring having 3 to 12 (i.e., C3-12 Cycloalkyl), preferably 3-10 (ie C 3-10 cycloalkyl), more preferably 3-8 ring atoms (ie C 3-8 cycloalkyl), most preferably 3, 4, 5 or 6 ring atoms (i.e. C 3-6 Cycloalkyl). Unless otherwise defined, a cycloalkyl group may be optionally substituted with one or more substituents. Preferably, the substituents of a cycloalkyl group may independently be hydroxy, alkyl, alkoxy, halogen, haloalkyl, amino, monoalkylamino, or dialkylamino. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like.

[0143] "Cycloalkyloxy" refers to a radical of the formula -OR, where R is a cycloalkyl radical as defined herein. Exemplary cycloalkyloxy radicals include cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like. "Cycloalkylalkyl" or "cycloalkylalkylene" refers to an -alkylene(cycloalkyl) radical, where cycloalkyl and alkylene are as defined above. "Cycloalkylalkyl" or "cycloalkylalkylene" is bonded to the parent molecular structure through an alkyl(alkylene) radical.

[0144] "Heteroaromatic ring system" or "heteroaromatic ring" refers to a monocyclic (e.g., 5- or 6-membered), bicyclic (6- to 12-membered), or polycyclic ring system in which at least one ring is aromatic and contains at least one heteroatom (e.g., N, O, or S) as a ring atom, and the remaining ring atoms are carbon. In some cases, the aromatic ring containing at least one heteroatom contains 1, 2, 3, or 4 heteroatoms in the ring. In addition to the aromatic ring containing at least one heteroatom as a ring atom, the remaining rings in the "heteroaromatic ring system" or "heteroaromatic ring" can be saturated, partially unsaturated, or fully unsaturated rings.

[0145] "Heteroaryl", alone or as part of another group, refers to a monovalent radical of a "heteroaromatic ring system" or "heteroaromatic ring" as defined above. The point of attachment of a heteroaryl group shall be on the aromatic ring. Examples of heteroaryl groups include, but are not limited to, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl, thiadiazolyl, pyrazinyl, thienyl, furanyl, pyranyl, pyridinyl, pyrrolyl, pyrazolyl, pyrimidinyl, quinolyl, isoquinolyl, benzofuranyl, benzofuranyl, benzothienyl, benzothiopyranyl, benzimidazolyl, benzoxazolyl, benzoxadiazolyl, benzothiazolyl, benzothiadiazolyl, benzopyranyl, indolyl, isoindolyl, triazolyl, triazinyl, quinoxalinyl, purinyl, quinazolinyl, quinolizinyl, naphthyridinyl, pteridinyl, carbazolyl, aza Base, diazepine A heteroarylene group refers to a heteroaryl group as defined above with two attachment sites. Unless otherwise defined, heteroaryl groups include substituted or unsubstituted forms.

[0146] "Heterocyclic ring system" or "heterocycle" refers to monocyclic, bicyclic, and polycyclic ring systems in which at least one ring is saturated or partially unsaturated (but nonaromatic) and contains at least one heteroatom as a ring atom. The heterocyclic ring system or heterocycle may be attached to a pendant group at any heteroatom or carbon atom that creates a stable structure and any ring atom may be optionally substituted.

[0147] "Heterocyclyl" refers to a heterocyclic ring system or a monovalent radical of a heterocycle as defined above, generally a stable monocyclic ring (e.g., 3-8 members, i.e., 3-, 4-, 5-, 6-, 7-, or 8-membered) or bicyclic ring (e.g., 5-12 members, i.e., 5-, 6-, 7-, 8-, 9-, 10-, 11-, or 12-membered) or polycyclic ring (e.g., 7-14 members, i.e., 7-, 8-, 9-, 10-, 11-, 12-, 13-, or 14-membered), including fused, spiro, and / or bridged ring structures, which is saturated, partially unsaturated, and which contains carbon atoms and 1, 2, 3, or 4 heteroatoms independently selected from N, O, and S as ring atoms. The heterocyclyl is preferably a 3- to 14-membered heterocyclyl, more preferably a 3- to 8-membered heterocyclyl, and most preferably a 4- to 6-membered heterocyclyl. Representative heterocyclyl groups include ring systems in which (1) each ring is non-aromatic and at least one ring contains a heteroatom, for example, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, pyrrolidinyl, pyrrolidonyl, piperidinyl, pyrrolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl; (2) at least one ring is non-aromatic. and contains heteroatoms as ring atoms and at least one other ring is an aromatic carbocyclic ring, for example, 1,2,3,4-tetrahydroquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl; and (3) at least one ring is non-aromatic and contains heteroatoms and at least one other ring is aromatic and contains heteroatoms, for example, 3,4-dihydro-1H-pyrano[4,3-c]pyridine and 1,2,3,4-tetrahydro-2,6-naphthyridine. Heterocyclylene refers to a heterocyclyl as defined above with two attachment sites. In the present invention, preferably, the heterocyclylene is a bicyclic ring, one of which is a heteroaryl group and is connected to the other parts of the general formula through the heteroaryl group. In the present invention, preferably, the heterocyclylene is a 5-6 membered monocyclic heterocyclylene or an 8-10 membered bicyclic heterocyclylene. Unless otherwise defined, "heterocyclyl" and "heterocyclylene" include substituted or unsubstituted forms. When the heterocyclic group is saturated, the heterocyclic group may also be referred to as a heterocycloalkyl group.

[0148] "Heterocyclylalkyl" refers to an alkyl group substituted with a heterocyclyl group, wherein the heterocyclyl group and the alkyl group are as defined above.

[0149] The "alkylamino group" refers to a group having an alkyl-NR- structure, wherein R is H, or an alkyl group, a cycloalkyl group, an aryl group, a heteroaryl group, or the like as described above.

[0150] "Cycloalkylamino" refers to a group of the formula -N(R a )R b group, where R a is H, alkyl as defined herein or cycloalkyl as defined herein, R b is a cycloalkyl group as defined herein, or R a and R b Together with the nitrogen atom to which it is attached, it forms a 3-10 membered N-containing monocyclic or bicyclic heterocyclic group, such as tetrahydropyrrolyl. As used in the present invention, C3-C8 cycloalkylamino refers to an amino group containing 3-8 carbon atoms.

[0151] In the present invention, "ester group" refers to a group having a -C(O)-OR or RC(O)-O- structure, wherein R independently represents hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, or heterocyclic group, as defined above.

[0152] In the present invention, the term "amido" refers to a group having the structure -CONRR', wherein R and R' can independently represent hydrogen, alkyl or substituted alkyl, cycloalkyl or substituted cycloalkyl, aryl or substituted aryl, heterocycle or substituted heterocycle, as defined above. R and R' can be the same or different in the dialkylamine moiety.

[0153] In the present invention, the term "sulfonamide" refers to a group having the structure -SO2NRR', wherein R and R' can independently represent hydrogen, alkyl or substituted alkyl, cycloalkyl or substituted cycloalkyl, aryl or substituted aryl, heterocycle or substituted heterocycle, as defined above. R and R' can be the same or different in the dialkylamine moiety.

[0154] "Ketocarbonyl" refers to RC(=O)-, where R is alkyl, cycloalkyl, etc. as described above.

[0155] When a substituent is non-terminal, it is a substituent of the corresponding group, for example, alkyl for alkylene, cycloalkyl for cycloalkylene, heterocyclyl for heterocyclylene, alkoxy for alkyleneoxy, and the like.

[0156] In the present invention, each of the above-mentioned alkyl, alkoxy, cycloalkyl, heteroalkyl, aryl, heteroaryl, cycloheteroalkyl, alkenyl, alkyne, heterocycle, heterocyclyl, etc. may be substituted or unsubstituted.

[0157] In the present invention, the term "substituted" means that one or more hydrogen atoms on a specific group are replaced by a specific substituent. The specific substituent is the substituent described above, or the substituent appearing in the embodiments. Unless otherwise specified, a substituted group may have a substituent selected from a specific group at any substitutable site of the group, and the substituent may be the same or different at each position. It will be understood by those skilled in the art that the combination of substituents contemplated by the present invention is a combination that is stable or chemically feasible. Typical substitutions include, but are not limited to, one or more of the following groups: such as hydrogen, deuterium, halogen (e.g., a monohalogen substituent or a polyhalogen substituent, the latter such as a trifluoromethyl group or an alkyl group containing Cl3), cyano, nitro, oxo (such as =O), trifluoromethyl, trifluoromethoxy, cycloalkyl, alkenyl, alkynyl, heterocycle, aromatic ring, OR a SR a 、S(=O)R e 、S(=O)2R e 、P(=O)2R e 、S(=O)2OR e , P(=O)2OR e NR b R c NR b S(=O)2R e NR b P(=O)2R e 、S(=O)2NR b R c 、P(=O)2NR b R c 、C(=O)OR d 、C(=O)R a 、C(=O)NR b R c 、OC(=O)R a 、OC(=O)NR b R c NR b C(=O)OR e NR d C(=O)NR b R c NR d S(=O)2NR b R c NR d P(=O)2NR b R c NR b C(=O)R a , or NR b P(=O)2R e , where Ra R may independently represent hydrogen, deuterium, alkyl, cycloalkyl, alkenyl, alkynyl, heterocyclic or aromatic rings. b 、R c and R d may independently represent hydrogen, deuterium, an alkyl group, a cycloalkyl group, a heterocyclic ring or an aromatic ring, or R b and R c Together with the N atom, it can form a heterocyclic ring; R e and alkyl, cycloalkyl, alkenyl, alkynyl, heterocyclic ring or aromatic ring. The above typical substituents, such as alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocyclic ring or aromatic ring, may be optionally substituted. The substituents include, but are not limited to, halogen, hydroxyl, cyano, carboxyl (-COOH), C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3-12 membered heterocyclic group, aryl, heteroaryl, C1-C8 aldehyde group, C2-C 10 Acyl, C2-C 10 Ester group, amino group, C1-C6 alkoxy group, C1-C 10 sulfonyl, and C1-C6 urea group, etc.

[0158] "Cyano" refers to -CN.

[0159] "Nitro" refers to -NO2.

[0160] "Hydroxyl" refers to -OH.

[0161] "Amino" refers to -NH2 or RNH-, where R is ketocarbonyl, sulfonyl, sulfonamide, R a -C(=O)-, R a R b NC(=O)- etc., where R a and R b is an alkyl group, a cycloalkyl group, an aryl group or a heteroaryl group.

[0162] "Halogen" refers to any halogen radical, for example, -F, -Cl, -Br, or -I.

[0163] "Deuterated compound" refers to a compound in which one or more hydrogen atoms (H) are replaced by deuterium atoms (D).

[0164] In the present invention, the term "plurality" independently refers to 2, 3, 4, or 5.

[0165] The structural formula of the carbamate group is -NH-C(=O)-OR, wherein R is an alkyl group, an aryl group, a heteroaryl group, or the like.

[0166] Active ingredient

[0167] As used herein, the terms "compound of the present invention" or "active ingredient of the present invention" are used interchangeably to refer to a compound of Formula I, or a pharmaceutically acceptable salt, hydrate, solvate, isotope compound (such as a deuterated compound or deuterated substance), or prodrug thereof. The term also includes racemates and optical isomers.

[0168] The compound of formula I has the following structure:

[0169] R1, R2, R3, R4, X, X1, m and n are as defined above.

[0170] In another preferred embodiment, R1, R2, R3, R4, X, X1, m and n are each independently a corresponding specific group in the specific compound described in the present invention, the example compound or the compound shown in Table A.

[0171] The salts that may be formed by the compounds of the present invention also fall within the scope of the present invention. Unless otherwise indicated, the compounds of the present invention are understood to include their salts. The term "salt" as used herein refers to acidic or basic salts formed with inorganic or organic acids and bases. In addition, when the compound of the present invention contains a basic fragment, it includes but is not limited to pyridine or imidazole, and contains an acidic fragment, including but not limited to carboxylic acid, the zwitterions ("inner salts") that may be formed are included within the scope of the term "salt". Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful, for example, in separation or purification steps during the preparation process. The compounds of the present invention may form salts, for example, compound I reacts with a certain amount, such as an equivalent amount, of an acid or base, salts out in a medium, or is obtained by freeze-drying in an aqueous solution.

[0172] The compounds of the present invention contain basic moieties, including but not limited to amines or pyridine or imidazole rings, which may form salts with organic or inorganic acids. Typical acids that can form salts include acetates (e.g., acetic acid or trihaloacetic acid, such as trifluoroacetic acid), adipates, alginate, ascorbate, aspartate, benzoate, benzenesulfonate, bisulfate, borate, butyrate, citrate, camphor, camphorsulfonate, cyclopentanepropionate, diglycolate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide,

[0014] Examples of the present invention include, for example, hydroxyethylsulfonates, lactates, maleates, methanesulfonates, naphthalenesulfonates (e.g., 2-naphthalenesulfonate), nicotinates, nitrates, oxalates, pectinates, persulfates, phenylpropionates (e.g., 3-phenylpropionate), phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates (e.g., with sulfuric acid), sulfonates, tartrates, thiocyanates, toluenesulfonates such as p-toluenesulfonate, dodecanoates, and the like.

[0173] Certain compounds of the present invention may contain acidic moieties, including but not limited to carboxylic acids, which may form salts with various organic or inorganic bases. Typical base-forming salts include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, and salts formed with organic bases (e.g., organic amines), such as benzathine, dicyclohexylamine, hepamine (salt formed with N,N-di(dehydroabietyl)ethylenediamine), N-methyl-D-glucamine, N-methyl-D-glucamide, tert-butylamine, and salts formed with amino acids such as arginine, lysine, and the like. Basic nitrogen-containing groups can react with halide quaternary ammonium salts, such as small molecular alkyl halides (such as chlorides, bromides and iodides of methyl, ethyl, propyl and butyl), dialkyl sulfates (such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate and dipentyl sulfate), long chain halides (such as chlorides, bromides and iodides of decyl, dodecyl, tetradecyl and tetradecyl), aralkyl halides (such as benzyl and phenyl bromide), etc.

[0174] Prodrugs and solvates (or solvates) of the compounds of the present invention are also encompassed within the scope.

[0175] The term "prodrug" herein refers to a compound that undergoes chemical transformation by metabolic or chemical processes to produce a compound, salt, or solvate of the present invention when treating a related disease. The compounds of the present invention include solvates, such as hydrates.

[0176] The compounds, salts or solvates of the present invention may exist in tautomeric forms (such as amides and imino ethers). All such tautomers are part of the present invention.

[0177] All stereoisomers of the compounds (e.g., those that may exist due to asymmetric carbon atoms for various substitutions), including enantiomeric and diastereomeric forms, are contemplated by the present invention. Individual stereoisomers of the compounds of the present invention may not exist simultaneously with other isomers (e.g., as a pure or substantially pure optical isomer having a particular activity), or may be mixtures, such as racemates, or mixtures with all other stereoisomers or portions thereof. The chiral centers of the present invention have either S or R configurations, as defined by the 1974 recommendations of the International Union of Pure and Applied Chemistry (IUPAC). Racemic forms can be resolved by physical methods, such as fractional crystallization, or by crystallization of diastereomers derived from them, or by separation by chiral column chromatography. Individual optical isomers can be obtained from the racemate by suitable methods, including but not limited to conventional methods, such as salt formation with an optically active acid followed by recrystallization.

[0178] The compounds of the present invention, obtained by sequential preparation, isolation, and purification, are described in the text to a concentration of 90% or greater by weight, for example, 95% or greater, or 99% or greater ("very pure" compounds). Such "very pure" compounds of the present invention are also considered part of the present invention.

[0179] All configurational isomers of the compounds of the present invention are encompassed, whether in mixture, pure or very pure form. The definition of the compounds of the present invention includes both cis (Z) and trans (E) olefin isomers, as well as cis and trans isomers of carbocyclic and heterocyclic rings.

[0180] Throughout the specification, groups and substituents may be chosen to provide stable fragments and compounds.

[0181] Specific functional groups and chemical term definitions are detailed below. For the purposes of this invention, chemical elements are referred to in the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75 th The definitions of specific functional groups are consistent with those in "Organic Chemistry," Thomas Sorrell, University Science Books, Sausalito, 1999, which is incorporated by reference in its entirety.

[0182] Certain compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention encompasses all compounds, including their cis and trans isomers, R and S enantiomers, diastereomers, (D) isomers, (L) isomers, racemic mixtures, and other mixtures. Additionally, asymmetric carbon atoms may represent substituents, such as alkyl groups. All isomers and mixtures thereof are encompassed by the present invention.

[0183] According to the present invention, mixtures of isomers can contain various ratios of isomers. For example, mixtures containing only two isomers can have the following ratios: 50:50, 60:40, 70:30, 80:20, 90:10, 95:5, 96:4, 97:3, 98:2, 99:1, or 100:0. All ratios of isomers are within the scope of the present invention. Similar ratios, as well as ratios for more complex mixtures of isomers, are readily understood by those skilled in the art and are also within the scope of the present invention.

[0184] The present invention also includes isotopically labeled compounds that are equivalent to the original compounds disclosed herein. However, in practice, it is common for one or more atoms to be replaced by atoms having a different atomic mass or mass number. Examples of isotopes of the compounds of the present invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine isotopes, such as 2 H. 3 H. 13 C. 11 C. 14 C. 15 N. 18 O. 17 O. 31 P. 32 P. 35 S. 18 F and 36 The compounds of the present invention, or enantiomers, diastereomers, isomers, or pharmaceutically acceptable salts or solvates thereof, which contain isotopes or other isotopic atoms of the above compounds are within the scope of the present invention. Certain isotopically labeled compounds of the present invention, such as 3 H and 14 Radioisotopes of C are also included and are useful in tissue distribution experiments of drugs and substrates. 3 H and carbon-14, i.e. 14 C, their preparation and detection are relatively easy. It is the first choice among isotopes. In addition, heavier isotope substitutions such as deuterium, i.e. 2 H, due to its excellent metabolic stability, has advantages in certain therapeutics, such as increasing half-life in vivo or reducing dosage, and therefore, may be preferred in some cases. Isotopically labeled compounds can be prepared using conventional methods by replacing readily available isotopically labeled reagents with non-isotopic reagents using the protocols disclosed in the examples.

[0185] If a synthesis of a specific enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, followed by separation of the resulting diastereomeric mixture and removal of the chiral auxiliary to obtain the pure enantiomer. Alternatively, if the molecule contains a basic functional group, such as an amino acid, or an acidic functional group, such as a carboxyl group, diastereomeric salts can be formed with a suitable optically active acid or base, followed by separation by conventional means such as fractional crystallization or chromatography to obtain the pure enantiomer.

[0186] As described herein, the compounds of the present invention may be substituted with any number of substituents or functional groups to expand their scope. Generally, the term "substituted," whether preceded or followed by the term "optionally," in formulas of the present invention including substituents, refers to the replacement of a hydrogen radical with a substituent of the specified structure. When multiple positions in a particular structure are substituted with multiple substituents of the specified structure, the substituents may be the same or different at each position. As used herein, the term "substituted" includes all permissible substitutions in organic compounds. Broadly speaking, permissible substituents include acyclic, cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic organic compounds. For example, heteroatoms of nitrogen may be substituted with hydrogen or any of the permissible organic compounds described above to supplement their valences. Furthermore, the present invention is not intended to limit the permissible substitutions in any way to organic compounds. The present invention recognizes that combinations of substituents and variables are advantageous for providing stable compounds for the treatment of diseases. As used herein, the term "stable" refers to compounds that are stable and maintain the structural integrity of the compound over a period of time sufficient to be detected, preferably over a period of time sufficient to be effective, as used herein for the purposes described above.

[0187] The metabolites of the compounds and pharmaceutically acceptable salts thereof involved in the present application, as well as prodrugs that can be converted into the structures of the compounds and pharmaceutically acceptable salts involved in the present application and in vivo, are also included in the claims of the present application.

[0188] Preparation method

[0189] The following schemes and examples describe methods for preparing compounds of Formula I. Starting materials and intermediates were purchased from commercial sources, prepared by known procedures, or described otherwise. In some cases, the order in which the steps of the reaction schemes are performed may be altered to facilitate the reaction or to avoid unwanted side reaction products.

[0190] The preparation method of the compound of formula I of the present invention is described in more detail below, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combinations can be easily carried out by those skilled in the art.

[0191] Typically, in the preparation process, each reaction is usually carried out under the protection of an inert gas in a suitable solvent at a temperature of 0 to 150° C., and the reaction time is usually 2 to 24 hours.

[0192] The preferred preparation method is as follows:

[0193] method:

[0194] Step 1: In a solvent (1,4-dioxane, tetrahydrofuran, 1,2-dichloroethane), SM1 and S1 react at 70-120 degrees to generate by palladium-catalyzed coupling or nucleophilic substitution reaction.

[0195] Step 2: In a solvent (1,4-dioxane, tetrahydrofuran, toluene), SM2 and S2 undergo palladium-catalyzed coupling or nucleophilic substitution reaction under alkaline conditions to obtain product T

[0196] In the above formulae, R1, R2, R3, R4, X, X1, m and n are as defined above.

[0197] Unless otherwise specified, the above starting materials can be purchased from commercial sources or synthesized according to reported literature.

[0198] Pharmaceutical compositions and methods of administration

[0199] Because the compounds of the present invention possess excellent CDK2 / 4 protein kinase inhibitory activity, the compounds of the present invention, their various crystalline forms, pharmaceutically acceptable inorganic or organic salts, hydrates, or solvates, and pharmaceutical compositions containing the compounds of the present invention as the primary active ingredient can be used to prevent and / or treat CDK2 / 4-related diseases. According to prior art, the compounds of the present invention or pharmaceutical compositions containing the compounds of the present invention can be used to prevent and / or treat the following diseases: inflammation, cancer, cardiovascular disease, infection, immune disease, and metabolic disease.

[0200] The compounds of the present invention may be used in combination with other drugs known to treat or ameliorate similar conditions. When administered in combination, the original drug's route of administration and dosage may remain unchanged, while the compound of the present invention is administered simultaneously or subsequently. When the compounds of the present invention are administered concurrently with one or more other drugs, a pharmaceutical composition containing both the one or more known drugs and the compound of the present invention may be preferably used. Drug combinations also include administering the compounds of the present invention and one or more other known drugs during overlapping time periods. When the compounds of the present invention are administered in combination with one or more other drugs, the dosage of the compound of the present invention or the known drug may be lower than the dosage of each drug administered alone.

[0201] The drugs or active ingredients that can be used in combination with the compounds of formula I include, but are not limited to, PD-1 inhibitors (such as nivolumab, pembrolizumab, JS-001, SHR-120, BGB-A317, IBI-308, GLS-010, GB-226, STW204, HX008, HLX10, BAT1306, AK105, LZM 009 or biosimilars of the above drugs), PD-L1 inhibitors (such as durvalumab, atezolizumab, CS1001, KN035, HLX20, SHR-1316, BGB-A333, JS003, CS1003, KL-A167, F 520, GR1405, MSB2311 or biosimilars of the above drugs, etc.), CD20 antibodies (such as rituximab, obinutuzumab, ofatumumab, tositumomab, ibritumomab tiuxetan, etc.), CD47 antibodies (such as Hu5F9-G4, CC-90002, TTI-621, TTI-622, OSE-172, SRF-231, ALX-148, NI-1701, SHR-1603, IBI188, IMM01), ALK inhibitors (such as ceritinib, alectinib, brigatinib, lorlatinib, ocatinib), PI3K inhibitors (such as idelalisib, dactolisib, taselisib, Bup sarlisib, etc.), BTK inhibitors (such as ibrutinib, tirabrutinib, acalabrutinib, etc.), EGFR inhibitors (such as afatinib, gefitinib, erlotinib, lapatinib, dacomitinib, icotinib, canertinib, etc.), VEGFR inhibitors (such as sorafenib, pazopanib, rivatotinib, cabozantinib, sunitinib, donafenib, etc.), HDAC inhibitors (such as givinostat, droxinostat, entinostat, dacilast, tekdinib, etc.), CDK inhibitors (such as palbociclib, ribociclib, abemaciclib, lerociclib, etc.), MEK inhibitors (such as selumetinib, ribociclib, ribociclib, ribociclib, etc.),(AZD6244), Trametinib (GSK1120212), PD0325901, U0126, AS-703026, PD184352 (CI-1040), etc.), Akt inhibitors (such as MK-2206, Ipatasertib, Capivasertib, Afuresertib, Uprosertib, etc.), mTOR inhibitors (such as Visusertib, etc.), SHP2 inhibitors (such as RMC-4630, JAB-3068, TNO155, etc.), IGF-1R inhibitors (such as Ceritinib, Ocatinib, Linsitinib, BMS-754807, etc.), , GSK1838705A, etc.), ER antagonists or degraders (such as tamoxifen, fulvestrant, etc.), aromatase inhibitors (such as letrozole, etc.), BCL2 or BCL-XL inhibitors (such as ABT-199, ABT-263, etc.), Hedgehog inhibitors (such as vismodegib, cyclopamine, etc.), chemotherapy drugs (such as cisplatin, etoposide, topotecan, etc.), PARP inhibitors (such as Olaparib, Veliparib, Rucaparib, etc.), ATR / ATM inhibitors (such as Ceralasertib, Berzosertib, etc.) or a combination thereof.

[0202] The dosage forms of the pharmaceutical composition of the present invention include (but are not limited to): injection, tablet, capsule, aerosol, suppository, film, pill, external ointment, controlled release or sustained release or nano preparation.

[0203] The pharmaceutical composition of the present invention comprises a safe and effective amount of a compound of the present invention or a pharmacologically acceptable salt thereof, and a pharmacologically acceptable excipient or carrier. "Safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably 10-1000 mg of the compound of the present invention per dose. Preferably, "one dose" is one capsule or tablet.

[0204] "Pharmaceutically acceptable carriers" refer to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0205] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration routes include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0206] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0207] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions can be delayed in a certain portion of the digestive tract. Examples of useful encapsulating components are polymeric substances and waxes. If desired, the active compound can also be microencapsulated with one or more of the above-mentioned excipients.

[0208] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures thereof.

[0209] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0210] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0211] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0212] Dosage forms for topical administration of the compounds of this invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0213] The treatment method of the present invention can be used alone or in combination with other treatment methods or therapeutic drugs.

[0214] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is administered to a mammal (e.g., a human) in need of treatment, wherein the dosage is a pharmaceutically effective dosage. For a 60 kg human, the daily dosage is generally 1 to 2000 mg, preferably 50 to 1000 mg. Of course, the specific dosage will also take into account factors such as the route of administration and the patient's health condition, all of which are within the skill of a skilled physician.

[0215] The present invention also provides a method for preparing a pharmaceutical composition, comprising the steps of: mixing a pharmaceutically acceptable carrier with the compound of the present invention or its crystal form, pharmaceutically acceptable salt, hydrate or solvate, thereby forming a pharmaceutical composition.

[0216] The present invention also provides a treatment method, which comprises the steps of administering the compound of the present invention, or its crystal form, pharmaceutically acceptable salt, hydrate or solvate, or the pharmaceutical composition of the present invention to a subject in need of treatment, for inhibiting CDK2 / 4 protein kinase.

[0217] The main advantages of the present invention include

[0218] (1) The compounds of the present invention have excellent inhibitory ability against CDK2 / 4 protein kinases;

[0219] (2) The compounds of the present invention have lower toxic side effects;

[0220] (3) The compounds of the present invention have better pharmacodynamics and pharmacokinetic properties.

[0221] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0222] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those familiar to those skilled in the art. Furthermore, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0223] Preparation Example

[0224] Example 1: Synthesis of Compound T-1

[0225] The synthetic route of compound T-1 is as follows:

[0226] The specific steps are as follows:

[0227] Step 1:

[0228] Compound SM 1 (5.0 g, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (THF) in an ice-water bath. Solid cesium carbonate (17.1 g, 2.5 eq) was added, followed by 3-chloropropylamine hydrochloride (2.7 g, 1.0 eq) with stirring. After addition, the mixture was allowed to warm to room temperature. After TLC monitoring, the solid was removed by filtration. The filter cake was washed with an appropriate amount of EA and extracted with water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to yield SM 2 (4.7 g, 71.9%) as a yellow solid. 1 H NMR (400 MHz, chloroform-d) δ 7.35 (s, 1H), 6.81 (t, J = 2.0 Hz, 1H), 6.64 (dd, J = 12.0, 2.0 Hz, 1H), 3.67 (t, J = 4.0 Hz, 2H), 3.46 (q, J = 8.0 Hz, 2H), 2.16 (p, J = 8.0 Hz, H).

[0229] Step 2:

[0230] Compound SM 2 (4.3 g, 1.0 eq) was dissolved in a mixture of methanol and water (v:v = 4:1). Iron powder (3.9 g, 5.0 eq) and solid ammonium chloride (1.5 g, 2.0 eq) were added and the mixture was heated to reflux for reaction. TLC monitoring was performed. After completion of the reaction, the mixture was cooled to room temperature and filtered. The filter cake was washed with an appropriate amount of EA. The filtrate was concentrated under reduced pressure to remove most of the solvent. The mixture was then extracted with saturated sodium bicarbonate solution and EA. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain compound SM 3 (2.0 g, 51.4%).

[0231] Step 3:

[0232] Compound SM 3 (1.3 g, 1.0 eq) and CDI (2.2 g, 2.5 eq) were dissolved in ultra-dry DMF solvent in sequence, and the temperature was raised to 50°C for reaction. After the reaction was monitored by TLC, the mixture was cooled to room temperature and extracted with EA and water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain compound SM 4 (1.2 g, 83.3%).

[0233] Step 4:

[0234] Compound SM 4 (4.0 g, 1.0 eq) was dissolved in phosphorus oxychloride (40 mL) and heated to 110°C for 24 hours. After completion of the reaction, the mixture was cooled to room temperature and slowly poured into ice water for quenching. The mixture was extracted with EA, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by column chromatography afforded compound SM 5 (137 mg, 24.9%).1 H NMR (400 MHz, chloroform-d) δ 6.98 (dd, J = 10.0, 1.7 Hz, 1H), 6.93 (d, J = 1.7 Hz, 1H), 3.93 (t, J = 6.2 Hz, 2H), 3.48-3.31 (m, 2H), 3.22 (s, 3H), 2.26 (p, J = 6.0 Hz, 2H).

[0235] Step 5:

[0236] Compound SM 5 (8.0 g, 1.0 eq) was dissolved in 4.0 M methylamine ethanol solution (80 mL) and heated to 80°C for 24 hours. The reaction was monitored by TLC. After completion of the reaction, the mixture was cooled to room temperature and concentrated under reduced pressure to remove the solvent. The mixture was then extracted with EA and water, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain compound SM 6 (3.5 g, 50.2%). 1 H NMR (400 MHz, chloroform-d) δ 6.98 (dd, J = 10.0, 1.7 Hz, 1H), 6.93 (d, J = 1.7 Hz, 1H), 3.93 (t, J = 6.2 Hz, 2H), 3.48-3.31 (m, 2H), 3.22 (s, 3H), 2.26 (p, J = 6.0 Hz, 2H).

[0237] Step 6:

[0238] Under nitrogen, compound SM 6 (0.26 g, 1.0 eq), pinacol diboronate (0.46 g, 2.0 eq), Pd(dppf)Cl2 (34 mg, 0.05 eq), and potassium acetate (0.27 g, 3.0 eq) were added sequentially to a dry 50 mL three-necked flask. The atmosphere was purged with nitrogen three times, and ultra-dry 1,4-dioxane was added. The reaction was heated to reflux. TLC monitoring was performed. After completion of the reaction, the mixture was cooled to room temperature and extracted with Ea and water. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by column chromatography afforded compound SM7 (0.21 g, 69.3%). LCMS: [M+H] + =332.6.

[0239] Step 7:

[0240] Under nitrogen, compound SM7 (0.21 g, 1.0 eq), 2,4,5-trichloropyrimidine (0.21 g, 2.0 eq), tetrakistriphenylphosphine palladium (71.5 mg, 0.1 eq), and solid sodium carbonate (0.2 g, 3.0 eq) were added to a 50 mL three-necked flask. A mixture of 1,4-dioxane and water (v:v = 10:3) was added. The atmosphere was replaced with nitrogen three times, and the temperature was raised to reflux for reaction. TLC monitoring was performed. After completion of the reaction, the mixture was cooled to room temperature and extracted with Ea and water. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by column chromatography afforded compound SM8 (0.13 g, 59.2%). LCMS: [M+H] + =352.5.

[0241] Step 8:

[0242] Compound SM8 (120 mg, 1.0 eq), pyrazolo[1,5-a]pyridin-5-amine (54.5 mg, 1.2 eq), palladium acetate (7.7 mg, 0.1 eq), BINAP (42.5 mg, 0.2 eq), and cesium carbonate (333.2 mg, 3.0 eq) were added sequentially to a 25 mL three-necked flask. 1,4-Dioxane was added, the atmosphere was replaced with nitrogen, and the temperature was raised to reflux. The reaction was monitored by TLC. After completion of the reaction, the mixture was cooled to room temperature and extracted with EA and water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to yield compound T-1 (70 mg, 45.8%). 1 H NMR (400MHz, DMSO-d6) δ10.24(s,1H),8.68(s,1H),8.58(d,J=8.0Hz,1H),8.32 (d,J=2.3Hz,1H),7.88(d,J=2.0Hz,1H),7.58(d,J=1.2Hz,1H),7.45(dd,J=12. 0,1.2Hz,1H),7.09(dd,J=7.5,2.4Hz,1H),6.41(d,J=2.2Hz,1H),4.06(t,J=6. 0Hz,2H),3.41(t,J=6.0Hz,2H),3.14(s,3H),2.19(t,J=5.9Hz,2H).LC-MS[M+H] + =449.3.

[0243] Example 2-43: Synthesis of Compounds T-2 to T-43

[0244] Referring to the synthesis method of Example 1, the corresponding raw materials were used to synthesize the compounds shown in the following table:

[0245] Example 2: Synthesis of Compound T-81

[0246] The synthetic route is as follows:

[0247] The experimental process is as follows:

[0248] Step 1: Dissolve compound SM 1 (25.0 g, 1.0 eq) and DMF (2.16 mL, 0.1 eq) in 500 mL of dry dichloromethane. Cool to 0°C in an ice-water bath, then slowly add SOCl 2 (30.5 mL, 1.5 eq) dropwise. After the addition is complete, heat to 40°C and react for 3 hours. Return to room temperature to allow the reaction to precipitate a white solid. After TLC monitoring, concentrate under reduced pressure to remove most of the solvent. Filter to obtain a white solid cake. Wash the cake with ethyl acetate and petroleum ether, then dry it to yield SM 2 (34 g, 84.2%). 1 H NMR (400 MHz, deuterated methanol) δ 3.81-3.62 (m, 2H), 3.52 (m, 1H), 2.16 (m, 1H), 2.02 (m, 1H), 1.34 (d, J = 6.6 Hz, 3H).

[0249] Step 2: Compound SM 3 (56.0 g, 1.0 eq) was dissolved in 500 mL of ultra-dry DMF. Solid potassium carbonate (97.6 g, 3.0 eq) was added, and SM 2 (34.0 g, 1.0 eq) was added with stirring. After the addition was complete, the temperature was raised to 80°C for reaction. After completion of the reaction, the solid was removed by filtration. The filter cake was washed with an appropriate amount of EA and extracted with water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to yield SM 4 (58.5 g, 76.5%) as a yellow solid.

[0250] Step 3: Compound SM 4 (58.5 g, 1.0 eq) was dissolved in a mixture of ethanol and water (v:v = 3:1). Iron powder (60.3 g, 6.0 eq) and solid ammonium chloride (19.2 g, 2.0 eq) were added. The reaction was heated to reflux under nitrogen for 1 hour. TLC monitoring was performed. After completion of the reaction, the mixture was cooled to room temperature and filtered. The filter cake was washed with an appropriate amount of EA. The filtrate was concentrated under reduced pressure to remove most of the solvent. The mixture was then extracted with saturated sodium bicarbonate solution and EA. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain compound SM 5 (48.4 g, 91.7%).

[0251] Step 4: Compound SM 5 (48.0 g, 1.0 eq) and CDI (66.3 g, 2.5 eq) were sequentially dissolved in 720 mL of ultra-dry DMF solvent, and the temperature was raised to 50°C for reaction. After the reaction was completed by TLC monitoring, the mixture was cooled to room temperature and extracted with EA and water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain compound SM 6 (48.0 g, 91.9%).

[0252] Step 5: Compound SM 6 (48 g, 1.0 eq) was added to 167 mL of phosphorus oxychloride and heated to 100°C for 18 hours. After completion of the reaction, the mixture was cooled to room temperature and slowly poured into ice water for quenching. The mixture was extracted with EA, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain compound SM 7 (45.5 g, 89.7%).

[0253] Step 6: To a 100 mL sealed jar, SM 7 (1.0 g, 1.0 eq), 2.0 M ethanolic methylamine solution (10 mL, 10.0 V), sodium iodide (2.2 g, 5.0 eq), and DMF (20 mL, 20 V) were added. The mixture was sealed and heated to 70°C for 12 hours. After completion of the reaction, the mixture was cooled to room temperature and extracted with EA and water. The organic phase was washed three times with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The organic phase was purified by silica gel column chromatography to obtain the desired product, SM 8 (400 mg, 47.8%). LCMS: [M+H] = 284.1, 286.1.

[0254] Step 7: Under nitrogen, add 60% NaH solid powder (35.6 mg, 1.1 eq) to 15 mL of ultra-dry THF. Place in an ice-water bath and add SM 8. After half an hour, add a solution of deuterated iodomethane in THF (117 mg, 1.0 eq) dropwise. Incubate for 2 hours. After TLC monitoring, quench the reaction with ice water. Extract with saturated ammonium chloride and EA. The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purify by silica gel column chromatography to obtain the target compound SM 9 (200 mg, 82.3%). LCMS: [M+H]+ = 301.2, 303.2.

[0255] Step 8: Under nitrogen, compound SM 9 (240 mg, 1.0 eq), pinacol diboronate (406 mg, 2.0 eq), Pd(dppf)Cl2 (29 mg, 0.05 eq), and potassium acetate (235 mg, 3.0 eq) were added sequentially to a dry 100 mL three-necked flask. The atmosphere was purged with nitrogen three times, and 10 mL of ultra-dry 1,4-dioxane was added. The reaction was heated to 110°C and monitored by TLC. After completion of the reaction, the mixture was cooled to room temperature and extracted with EA and water. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by column chromatography afforded compound SM 10 (200 mg, 72.7%). LCMS: [M+H]+ = 349.4.

[0256] Step 9: Under nitrogen, compound SM10 (220 mg, 1.0 eq), 2,4,5-trichloropyrimidine (0.11 mL, 1.5 eq), tetrakistriphenylphosphine palladium (36 mg, 0.05 eq), and solid sodium carbonate (201 mg, 3.0 eq) were added to a 50 mL three-necked flask. A mixture of 1,4-dioxane and water (v:v = 10:3) was added. The atmosphere was replaced with nitrogen three times, and the temperature was raised to 110°C for reaction. TLC monitoring was performed. After completion of the reaction, the mixture was cooled to room temperature and extracted with Ea and water. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by column chromatography afforded compound SM11 (100 mg, 42.9%). LCMS: [M+H]+ = 369.3.

[0257] Step 10: Compound SM-11 (100 mg, 1.0 eq), pyrazolo[1,5-a]pyridin-5-amine (43.3 mg, 1.2 eq), palladium acetate (6.1 mg, 0.1 eq), BINAP (33.7 mg, 0.2 eq), and cesium carbonate (264.4 mg, 3.0 eq) were added to a 25 mL three-necked flask. 1,4-dioxane was added, the atmosphere was replaced with nitrogen, and the temperature was raised to reflux for reaction. TLC monitoring was performed. After completion of the reaction, the mixture was cooled to room temperature and extracted with EA and water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain compound T-81. LCMS: [M+H]+ = 466.3. 1H NMR (400 MHz, chloroform-d) δ 8.40 (s, 1H), 8.30 (d,J=7.5Hz,1H),8.10(s,1H),7.83(s,1H),7.49(d,J=11.9Hz,1H),7.45(s,1H),7.30(s,1H),6.75(s,1H),6.30(s,1H),4.57-4.3 9(m,1H),3.54(d,J=3.2Hz,1H),3.24(d,J=7.5Hz,1H),2.34(tt,J=12.3,5.0Hz,1H),1.95(d,J=16.3Hz,1H),1.46(d,J=6.6Hz,3H).

[0258] Test Example 1 Enzyme activity test

[0259] The following biological activity tests were conducted on some of the compounds in the above examples and comparative examples.

[0260] The experimental process of biological activity test is as follows:

[0261] 1. Kinase activity test:

[0262] The test compounds were subjected to CDK2 and CDK4 kinase IC 50 Value detection.

[0263] (1) Reagent information

[0264] (2) Equipment information

[0265] (3) Research design

[0266] (1) Compound preparation:

[0267] ① Prepare the test compound into a 0.5 mM DMSO solution, and simultaneously prepare the positive control drug palbociclib into a 0.5 mM DMSO solution.

[0268] ②Three-fold dilution was performed to obtain 10 compound solutions with different concentrations.

[0269] (2) Enzyme assay:

[0270] ① Prepare a 1.3x enzyme solution containing enzyme, substrate, and cofactors as shown below.

[0271] ② Add 15 μL of 1.3x enzyme solution to each well and incubate at room temperature for 30 minutes.

[0272] ③ Add 5 μL of 4x ATP solution to start the reaction. Each test well contains the components listed in the table, and the final volume is 20 μL.

[0273] ④ Incubate for 60 minutes, then add 75 μL of buffer (containing 0.5 M EDTA) to stop the reaction.

[0274] ⑤ Use EZ to read the data of each test hole for analysis.

[0275] (3) Data analysis:

[0276] The percent inhibition was calculated using the concern ratio (CR) according to the following formula:

[0277] Wells treated with DMSO were used as positive controls, and wells without enzyme were used as negative controls.

[0278] % (percent inhibition) = 100 - 100*((CRPC-CRSample) / (CRPC-CRNC)).

[0279] Through the above test, the inhibitory activity IC of the test sample on CDK2 and CDK4 kinases was obtained. 50 The (nM) values ​​are shown in Table 1.

[0280] Table 1

[0281] As can be seen from the above table, through in vitro biological activity screening, using palbociclib as a reference substance, the compounds synthesized in the present application have a good inhibitory ability against CDK2 / 4 kinases and are expected to be further developed into drugs for regulating CDK2 / 4 protein kinase activity or treating CDK2 / 4-related diseases.

[0282] Test Example 2 Cell Anti-Proliferation Experiment

[0283] 1. Experimental materials and equipment:

[0284] Human breast cancer MCF-7 cells, ovarian cancer cells A2780, and human ovarian adenocarcinoma cells OVCAR3 were cultured in DMEM (Bio-Channel), DMSO (dimethyl sulfoxide), MTT (methylthiazolyl blue), 0.25% EDTA-Tripsin (trypsin digestion solution), 1x PBS (phosphate buffered saline, pH 7.2), 96-well plates (Corning), fetal bovine serum (FBS), 10,000 U / mL penicillin-G / streptomycin, a high-speed refrigerated centrifuge (EPPENDORF 5810R), and an enzyme-linked immunosorbent assay (Tecan Spark).

[0285] 2. Experimental preparation:

[0286] 1. Cell plating

[0287] A) Tumor cells were cultured in DMEM (high glucose, containing 10% FBS and 100 U / mL penicillin-G / streptomycin) at 37°C, 5% CO2 and saturated humidity to 80-90% confluence.

[0288] B) Remove the culture medium from the 10 cm culture dish;

[0289] C) Rinse the cells once with 10 ml of 1x PBS;

[0290] D) Add 4 ml of 0.25% EDTA-Tripsin and place in a 37°C, 5% CO2 incubator for trypsinization for 5 minutes. Transfer the tube to a 15 ml centrifuge tube and centrifuge at 200 g for 5 minutes. Discard the supernatant to obtain the cell pellet.

[0291] E) Resuspend in 4 ml of DMEM medium, count and adjust to 50,000 cells / ml.

[0292] F) The cell suspension was added to a 96-well plate at a volume of 100 μL per well and cultured overnight in a 37° C., 5% CO 2 incubator.

[0293] 2. Compound treatment

[0294] Compound dilution

[0295] A) Prepare test compound serial dilutions: Prepare a 1 mM stock solution of the test compound. Dissolve 1.5 μl of the stock solution in 1.5 ml of DMSO-free culture medium. Perform a 3-fold serial dilution in 0.1% DMSO culture medium for a total of 9 concentrations. The compound concentrations after dilution are as follows:

[0296] 333.33nM, 111.11nM, 37.03nM, 12.35nM, 4.15nM, 1.37nM, 0.46nM, 0.15nM

[0297] B) After thorough mixing, 100 μL of the culture compound solution was taken to replace the culture medium in the cell culture plate, with 4 replicates per concentration;

[0298] C) The cells were transferred to an incubator and incubated for 5 days.

[0299] 3. MTT assay

[0300] A) Remove the cell culture plate and add 10 μL of 5 mg / ml MTT in a biosafety cabinet;

[0301] B) Return the cell culture plate to the incubator and continue incubating for 3 hours;

[0302] C) Remove the cell culture plate and remove the culture medium. Add 100 μL of isopropanol (containing 0.4 mM HCl and 0.1% NP-40) and shake at room temperature for 30 minutes.

[0303] D) The absorbance value was measured on a TECAN enzyme-linked immunosorbent assay (ELISA) reader at a wavelength of 570 nm.

[0304] 4. Data Analysis

[0305] The following formula was used to calculate the cell viability (% Cell Viability):

[0306] %Cell Viability=100%×(Lum_Sample-Lum_LC) / (Lum_HC-Lum_LC)

[0307] Lum_HC: 0.1% DMSO control group cell readings

[0308] Lum_Sample: Cell readout with added compound

[0309] Lum_LC: Blank culture medium reading

[0310] The IC was obtained by curve fitting using GraphPad Prism 8 software. 50 Numerical value (unit: nM).

[0311] As shown in Table 2.

[0312] Table 2

[0313] As shown in Table 2, the compounds of the present invention have excellent anti-proliferative activity against breast cancer cells, ovarian cancer cells and human ovarian adenocarcinoma cells.

[0314] Experimental Example 3 Preclinical Pharmacokinetic Study in Rats

[0315] 1. Experimental materials and equipment:

[0316] Healthy adult male Sprague-Dawley rats, 6-8 weeks old, weighing 200-300 g, were purchased from XX Company. EDTA-Na2 anticoagulant, analytical balance, animal weighing scale, magnetic stirrer, refrigerated centrifuge, and single-channel manual pipette were also used.

[0317] 2. Experimental process:

[0318] 1. Drug preparation

[0319] Accurately weigh approximately 10 mg of the sample to be tested, dissolve it in 5% DMSO, then add 10% solutol HS-15 and 85% saline, sonicate, and vortex to mix to obtain a 1 mg / mL solution. Prepare freshly before use.

[0320] A 0.2 mL sample was taken into a 1.5 mL centrifuge tube and stored at -80°C for concentration analysis of the dosing solution.

[0321] 2. Animal Preparation

[0322] Animals were housed in rat cages and fasted for at least 10 hours, but not water, starting the day before the experiment. On the day of the experiment, each animal was weighed and tail-marked. Blank blood was collected from each animal before dosing. Blood was collected from the tail vein.

[0323] 3. Administer medication

[0324] Route of administration: oral administration (po)

[0325] Dosage: 10 mg / kg

[0326] Dosing volume: 10mL / kg

[0327] Operation procedure: Use your left hand wearing a bite-proof glove to catch the rat, make it stand upright, insert a No. 16 gavage needle into the mouth and throat, and try to insert the needle when there is no obvious resistance, and then inject the drug into the stomach.

[0328] 4. Sample collection

[0329] Before administration and at 0.5, 1, 2, 4, 6, 8, 12, and 24 hours after administration, 0.1 ml of whole blood was collected from the animals in EDTA-Na2 anticoagulant tubes. The tubes were inverted 3-4 times to mix thoroughly, and the plasma was separated by centrifugation at 10,000 g for 5 minutes at 4°C. The plasma was then stored at -80°C until testing. Blood was collected from the tail vein.

[0330] To perform the procedure, secure the rat in a fixture, fully exposing its tail. Wipe the tail with alcohol, allowing the surface skin to absorb the alcohol and significantly dilate the veins. Select a suitable vein on either side and insert the needle approximately one-third of the way from the tip of the tail. Use an insulin syringe with the needle facing upward. Once the skin is punctured, the needle should immediately become parallel. You should feel minimal resistance to the needle sliding through the vein and blood should return to the syringe, indicating entry. Draw approximately 0.1-0.2 ml of whole blood. Remove the needle and apply pressure to stop the bleeding.

[0331] 3. Sample analysis:

[0332] Preparation of standard curve: 25 μL of blank rat plasma was taken into centrifuge tubes, 25 μL of prepared standard series solution (prepared in methanol) was added, and then 200 μL of internal standard solution (prepared in methanol) was added. The mixture was vortexed for 2 min and centrifuged at 10,000 g for 10 min at 4°C.

[0333] For unknown plasma sample preparation, aspirate 25 μL of drug-containing rat plasma, add 25 μL of methanol and 200 μL of internal standard solution, vortex mix for 2 minutes, and centrifuge at 10,000 g for 10 minutes at 4°C. The supernatant is analyzed by LC / MS / MS.

[0334] 4. Data Processing

[0335] A quantitative assay for the test compound was established using a Shimadzu liquid chromatography and Triple Quad™ 6500+AB mass spectrometer. The unchanged drug concentration in plasma was determined. Plasma concentration-time curves were plotted, and key pharmacokinetic parameters were calculated using a non-compartmental model in WinNonlin Phoenix software. The results are shown in Table 3.

[0336] Table 3

[0337] It can be seen from Table 3 that the compounds of the present invention all have excellent pharmacokinetic properties.

[0338] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A compound, characterized in that The compound is a compound of formula I, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, solvate, isotope compound or prodrug thereof, in: X is N or CR5; X1 is N or CR5; R1 is selected from the group consisting of H, halogen, cyano, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl; R2 is as shown in Formula Ia or Ib; in, W3, W4 and W5 are each independently selected from: CH, N, NH; One of W2 and W6 is C, and the other is C or N; R6 are each independently selected from the following group: halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 3-6 Cycloalkyl, -CO-C 1-6 Alkyl, -COO-C 1-6 Alkyl, 4 to 6 membered heterocyclic group, -C 1-4 Alkylene-C 3-6 Cycloalkyl, -C 1-4 wherein the alkyl, alkylene, cycloalkyl and heterocyclic groups are optionally substituted by one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy; The subscript p is 0, 1, or 2; W1 and W7 are each independently CH or N; Subscript o is 0 or 1; R7 are each independently selected from the following group: halogen, C 1-6 Alkyl, C 1-6 Haloalkyl; Subscript q is 0, 1, 2, or 3; R3 are each independently selected from the following group: H, C 1-6 Alkyl, C 3-6 Cycloalkyl, hydroxyl, amino, C 1-6 Alkoxy (such as methoxy), C 1-6 Haloalkyl (such as CF3), C 1-6 Haloalkoxy, C 1-6 Hydroxyalkyl, 5-8 membered heterocyclyl, 5-8 membered heteroaryl; or, two R3 located on the same atom together form an oxo group (=O); R4 is selected from the following group: H, C 1-6 Alkyl, -CO-C 1-6 Alkoxy, -COC 1-6 Alkyl, C 3-8 Cycloalkyl, 4-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S; wherein the alkyl, aryl, cycloalkyl and heterocycloalkyl are optionally substituted by one or more groups selected from the group consisting of halogen, cyano, hydroxyl, C 1-6 Alkyl, -CO-C 1-6 Alkyl, C 1-6 Alkoxy, amino, -COOH, -CONH2, -CF3C(F)2, optionally C 1-6 Alkyl, -CH(F)2, -COO-C 1-6 Alkyl or -S(O)2-C 6-10 Aryl substituted with 1, 2 or a 5-8 membered heterocycloalkyl group having 3 heteroatoms selected from N, O or S; R5 is selected from the group consisting of H, hydroxy, halogen, cyano, C 1-6 Alkyl, C 1-6 Haloalkyl, -CO-C 1-6 Alkoxy; The subscript m is selected from the following group: 0, 1, 2, 3, 4; Subscript n is 0, 1, 2, 3 or 4.

2. The method according to claim 1, characterized in that: X is N or CR5; X1 is N or CR5; R1 is selected from the group consisting of H, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl; R2 is as shown in Formula Ia or Ib; in, W3, W4 and W5 are each independently selected from: CH, N, NH; One of W2 and W6 is C, and the other is C or N; R6 are each independently selected from the following group: halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 3-6 Cycloalkyl, 4- to 6-membered heterocyclic group, -C 1-4 Alkylene-C 3-6 Cycloalkyl, -C 1-4 wherein the alkyl, alkylene, cycloalkyl and heterocyclic groups are optionally substituted by one or more substituents selected from the group consisting of halogen, hydroxyl, cyano, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy; The subscript p is 0, 1, or 2; W1 and W7 are each independently CH or N; Subscript o is 0 or 1; R7 are each independently selected from the following group: halogen, C 1-6 Alkyl, C 1-6 Haloalkyl; Subscript q is 0, 1, 2, or 3; R3 are each independently selected from the following group: H, C 1-6 Alkyl, C 3-6 Cycloalkyl, hydroxyl, C 1-6 Alkoxy; R4 is selected from the following group: H, C 1-6 Alkyl, -CO-C 1-6 Alkoxy, -COC 1-6 alkyl; R5 is selected from the following group: H, C 1-6 Alkyl, -CO-C 1-6 Alkoxy; The subscript m is selected from the following group: 0, 1, 2, 3, 4; Subscript n is 0, 1, 2, 3 or 4.

3. The compound according to claim 1 or 2, characterized in that R2 is as shown in Formula Ia.

4. The compound according to claim 1 or 2, characterized in that In formula (Ia), W1 is CH, W2 is C, W3 is NH, W4 is CH, W5 is N, W6 is C and W7 is CH; or, W1 is CH, W2 is C, W3 is CH, W4 is CH, W5 is N, W6 is N and W7 is CH; or, W1 is CH, W2 is C, W3 is N, W4 is CH, W5 is N, W6 is N and W7 is CH; or, W1 is CH, W2 is N, W3 is CH, W4 is CH, W5 is N, W6 is C and W7 is N; or, W1 is CH, W2 is CH, W3 is N, W4 is CH, W5 is CH, W6 is N and W7 is CH; or, W1 is CH, W2 is CH, W3 is N, W4 is N, W5 is CH, W6 is N and W7 is N; or, W1 is CH, W2 is C, W3 is NH, W4 is CH, W5 is N, W6 is C and W7 is N; or, W1 is CH, W2 is C, W3 is N, W4 is CH, W5 is N, W6 is N and W7 is N; or, W1 is CH, W2 is C, W3 is NH, W4 is CH, W5 is CH, W6 is C and W7 is CH; or, W1 is CH, W2 is C, W3 is NH, W4 is N, W5 is CH, W6 is C and W7 is CH; or, W1 is CH, W2 is C, W3 is NH, W4 is CH, W5 is N, W6 is C and W7 is CH; or, W1 is CH, W2 is C, W3 is CH, W4 is N, W5 is NH, W6 is C and W7 is CH; or, W1 is CH, W2 is C, W3 is N, W4 is N, W5 is NH, W6 is C and W7 is CH; or, W1 is CH, W2 is C, W3 is CH, W4 is NH, W5 is N, W6 is C and W7 is CH; or, W1 is N, W2 is C, W3 is CH, W4 is NH, W5 is N, W6 is N and W7 is CH; or, W1 is CH, W2 is C, W3 is CH, W4 is CH, W5 is N, W6 is N and W7 is N; In formula (Ib), W2 is C, W3 is CH, W4 is CH, W5 is N, W6 is N and o is 0 or 1.

5. The compound according to claim 1 or 2, characterized in that Select from the following group:

6. The compound according to claim 1 or 2, characterized in that Select from the following group:

7. The compound according to claim 1 or 2, characterized in that X is N, X1 is N, and subscript n is 0 or 1.

8. The compound according to claim 1 or 2, characterized in that The compounds have one or more of the following characteristics: R1 is selected from the following group: H, F, Cl, methyl, Br, CF3, R3 is selected from the group consisting of H, methyl, cyclopropyl, hydroxyl, methoxy; R4 is selected from the group consisting of H, methyl, ethyl, 9. The compound according to claim 1 or 2, characterized in that The isotope compound is a deuterated compound.

10. The compound according to claim 1, characterized in that The compound is selected from Table A: Table A 11. A pharmaceutical composition, characterized in that Contains a preventive and / or therapeutically effective amount of the compound according to any one of claims 1 to 10, and a pharmaceutically acceptable carrier.

12. Use of the compound according to claim 1 or the pharmaceutical composition according to claim 11, characterized in that: For preparing one or more drugs selected from the following group: (a) A drug used as a CDK2 / 4 protein kinase inhibitor; (b) drugs for modulating CDK2 / 4 protein kinase activity; (c) Drugs for preventing or treating CDK2 / 4 related diseases.

13. The use according to claim 12, characterized in that The CDK2 / 4 related disease is selected from the group consisting of inflammation, cancer, cardiovascular disease, infection, immune disease, and metabolic disease.

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