A compound as a BTK inhibitor and its preparation method and use

Abstract

A compound as a BTK inhibitor and its preparation method and use, the compound has the structure shown in formula I or its tautomer, mesomer, racemate, enantiomer, diastereomer or mixture form, pharmaceutically acceptable hydrate, solvate or salt; wherein: A1, A2, A3, A4, A5, A6 are independently selected from C-R5 or N; and at least one of these is N; M is selected from substituted or unsubstituted saturated hydrocarbon group or heterosaturated hydrocarbon group, substituted or unsubstituted unsaturated cyclic group or heterocyclic group, substituted or unsubstituted monocyclic, bicyclic or tricyclic aromatic group or heteroaromatic group. The BTK protein kinase inhibitor of the present invention has strong inhibitory effect on wild-type BTK and mutant BTK (C481S), and has good pharmacokinetic properties, and has good application prospects.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a compound used as a BTK protein kinase inhibitor, and a preparation method and use thereof. Background Art

[0002] Bruton's tyrosine kinase (BTK) is a member of the Tec family of non-receptor protein tyrosine kinases and is mainly expressed in a variety of hematopoietic cell lines. The Tec family is the second largest family of human non-receptor kinases after the Src family. Its main members include BTK, BMX (etk), ITK, TEC and TXK (RLK). In 1993, BTK was identified as a defective protein in human X-linked agammaglobulinemia (XLA). BTK is a key regulator of the B cell receptor (BCR) signal transduction pathway. It plays an important role in the activation, proliferation, differentiation and survival of B cells and is closely related to a variety of B cell tumors and autoimmune diseases.

[0003] The BTK structure contains five main domains, namely the PH domain (Pleckstrin homology), TH domain (Tec homology), SH3 domain (Src homology 3), SH2 domain (Src homology 2) and SH1 domain (Src homology 1). BTK activation (phosphorylation) initially occurs in the activation loop in the SH1 domain, and further activation occurs in the SH2 and SH3 domains containing the main autophosphorylation sites; these SH domains also contain the nuclear localization signal (NLS) and nuclear export sequence (NES) required for BTK to shuttle between the nucleus and the cytoplasm.

[0004] BTK plays an irreplaceable role in the generation of B lymphocytes. It can control the development and differentiation of B cells by activating positive cell cycle regulators and differentiation factors, and can also control the survival and proliferation of B cells by regulating the expression of pro-apoptotic and anti-apoptotic proteins. Sustained activation of BTK is a prerequisite for the development of chronic lymphocytic leukemia (CLL); abnormal BCR-BTK signaling promotes the survival of activated B cell subtypes in diffuse large B cell lymphoma (DLBCL). BTK gain-of-function mutations have also been confirmed in colorectal cancer, acute lymphocytic leukemia (ALL), and chronic myeloid leukemia (CML). It can be seen that abnormal activation of BTK-dependent pathways has been shown to be closely related to the occurrence and development of a variety of tumors.

[0005] Currently approved irreversible BTK inhibitors, such as ibrutinib, acalabrutinib, and zanubrutinib, can selectively form irreversible covalent bonds with the cysteine ​​residue (Cys-481) of BTK, inhibiting BTK activity to achieve the purpose of treating related diseases. However, some cancer patients will develop resistance to first-generation BTK inhibitors, resulting in new unmet clinical needs. Studies have shown that BTK-C481S mutation is one of the main resistance mechanisms associated with this, so drugs that can target and inhibit BTK-C481S mutants are expected to provide new treatment options. For example, ARQ-531 is an orally bioavailable, effective, and reversible dual inhibitor of wild-type and C481S mutant BTK; the initial clinical results of ARQ-531 have shown its effectiveness in patients with C481S mutant BTK. Summary of the invention

[0006] In view of this, the present application provides a compound as a BTK inhibitor and a preparation method and use thereof. The compound provided by the present invention can be used as a BTK protein kinase inhibitor and has the characteristics of high inhibitory activity and the like.

[0007] The present invention provides a compound having a structure shown in Formula I or its tautomer, mesomer, racemate, enantiomer, diastereomer or a mixture thereof, a pharmaceutically acceptable hydrate, solvate or salt:

[0008]

[0009] Wherein: A1, A2, A3, A4, A5, A6 are independently selected from C-R5 or nitrogen (N); and at least one of A1, A2, A3, A4, A5, A6 is N;

[0010] M is selected from substituted or unsubstituted saturated hydrocarbon or heterosaturated hydrocarbon, substituted or unsubstituted unsaturated cyclic or heterocyclic, substituted or unsubstituted monocyclic, bicyclic or tricyclic aromatic or heteroaromatic; wherein the substituted groups are independently selected from aromatic or heteroaromatic, alkyl or heteroalkyl, cycloalkyl or heterocycloalkyl, unsaturated cyclic or heterocyclic, phenoloxy, halogen, hydroxyl, cyano, amino, ester, nitro, sulfhydryl, amide, sulfonyl, phosphoryl, alkyloxyphosphino, alkylsulfone, alkylsulfoxide; further, the substituted groups are aromatic or heteroaromatic substituted by any groups, more preferably phenyl substituted by any groups;

[0011] Q is selected from CR 10 R 11 NR 12, oxygen (O), sulfur (S), S (O), S (O) 2;

[0012] R1, R2, R3, R4, R5, R 10 , R 11 , R 12 R3 and R4 are independently selected from hydrogen, deuterium, halogen, substituted or unsubstituted alkyl or heteroalkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted unsaturated cyclic or heterocyclic, substituted or unsubstituted aryl or heteroaryl, hydroxyl, cyano, amino, ester, nitro, sulfhydryl, amide, sulfonyl, phosphoryl, alkyl phosphine, alkyl sulfone, alkyl sulfoxide; or R3, R4 together with the carbon atom to which they are attached constitute a substituted or unsubstituted C3-C10 cycloalkyl or heterocycloalkyl; the substituted substituent is selected from halogen, hydroxyl, cyano, amino, sulfhydryl, nitro, carboxyl, hydroxyamino, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, aryl, heteroaryl, ester, acyl, amide, sulfonyl, phosphoryl;

[0013] m is selected from integers from 0 to 6; n is selected from integers from 0 to 3.

[0014] The compounds described in the present invention are in any form having the structure of Formula I, including tautomers, mesomers, racemates, enantiomers, diastereomers or mixtures thereof, pharmaceutically acceptable hydrates, solvates or salts, etc.

[0015] In the present application, "selected from" is generally a parallel relationship of or. In the structure shown in Formula I, preferably three or four of A1, A2, A3, A4, A5, and A6 are N; the position of R2 is not limited, preferably in the opposite position to R1. In the present application, the substitution may be a single substitution or a multiple substitution (for example, a disubstituted or trisubstituted), and the specific substitution position is not particularly limited. The unsubstituted saturated hydrocarbon group includes an unsubstituted alkyl group and an unsubstituted cycloalkyl group; the heterocyclic group, heteroaryl group and other groups are groups in which one or more carbon atoms can be replaced by heteroatoms, and the heteroatoms are atoms such as oxygen, sulfur, nitrogen, phosphorus (P) other than carbon (C). In addition, the above-mentioned halogens include fluorine (F), chlorine (Cl), bromine (Br), etc., preferably fluorine or chlorine. The above-mentioned "C3-C10" is an integer selected from 3 to 10 carbon atoms, and similar expressions are not repeated below.

[0016] In the present application, the ring system formed by the bridge atom connected to the ring by chemical bonds (as shown in the following formula) means that the bridge atom can be connected to any connectable C atom on the ring, that is, any spiro or bridged ring structure compound can be formed. For example, the following formula indicates that the bridge atom Q can be connected to any C atom that can connect to the bridge atom on the six-membered ring, that is, connected to the same C atom to form a spiro compound, such as the bridge atoms are all connected to the No. 2 C atom or are all connected to the No. 3 C atom, etc.; connected to different C atoms to form a bridged ring compound, such as the bridge atom is respectively connected to the No. 1 and No. 4 C atoms or the No. 2 and No. 4 C atoms, etc.;

[0017]

[0018] Preferably, the compound has the structure shown in Formula II or its tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, pharmaceutically acceptable hydrate, solvate or salt:

[0019]

[0020] Wherein, R1 is selected from hydrogen, halogen, hydroxyl, cyano, amino, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C1-C6 heteroalkyl, substituted or unsubstituted C3-C6 heterocycloalkyl; further, R1 is selected from hydrogen, amino, methyl, ethyl, methoxy, cyano, trifluoromethyl, isopropyl, cyclopropyl; further, R1 is selected from hydrogen (H), amino (NH2), methyl (CH3);

[0021] R2 is selected from hydrogen, halogen, hydroxyl, cyano, amino, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C1-C6 heteroalkyl, substituted or unsubstituted C3-C6 heterocycloalkyl; further, R2 is selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, methoxy, cyano, trifluoromethyl, isopropyl, cyclopropyl; further, R2 is selected from hydrogen, chlorine, methyl;

[0022] R3 and R4 are selected from hydrogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C1-C6 heteroalkyl, substituted or unsubstituted C3-C6 heterocycloalkyl; or R3 and R4 together with the carbon atom to which they are connected constitute a substituted or unsubstituted C3-C6 cycloalkyl or a heterocycloalkyl containing N or O atoms;

[0023] Further, R3 and R4 are selected from hydrogen, methyl, ethyl, isopropyl, cyclopropyl or R3, R4 together with the carbon atom to which they are attached form cyclopropyl, azetidinyl, azetyl, azetyl, oxetanyl, oxolyl, oxetanyl;

[0024] R6 is selected from hydrogen, halogen, hydroxyl, cyano, amino, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C1-C6 heteroalkyl, substituted or unsubstituted C3-C6 heterocycloalkyl; further, R6 is selected from hydrogen, halogen, cyano, substituted or unsubstituted C1~C3 alkyl, substituted or unsubstituted C1~C3 alkoxy; further, R6 is selected from hydrogen, fluorine, chlorine, bromine, trifluoromethyl, methyl, methoxy, trifluoromethoxy, difluoromethoxy; further R6 is hydrogen or fluorine.

[0025] m is selected from 0, 1, 2, 3; n is selected from 0, 1, 2; n1 is selected from 0, 1, 2, 3, 4;

[0026] R7 is selected from substituted or unsubstituted aryl, substituted or unsubstituted pyridyl, wherein the substituted substituents are independently selected from halogen, hydroxyl, amino, cyano, alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl; further, the substituents are independently selected from fluorine, chlorine, bromine, cyano, amino, C1-C3 alkyl, C1-C3 alkoxy, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl; further, the substituents are independently selected from fluorine, chlorine, bromine, cyano, trifluoromethyl, trifluoromethoxy, difluoromethoxy, methoxy, deuterated methoxy, cyclopropyl, cyclopropylmethoxy, ethyl, isopropyl, isobutyl; wherein the number of the substituents is an integer between 0 and 5;

[0027] X is selected from In some embodiments, X is Among them, R9, R 13 R9 and R 13 and the carbon atoms to which they are connected together constitute a substituted or unsubstituted C3-C6 cycloalkyl or a substituted or unsubstituted C3-C6 heterocycloalkyl containing N or O; further, independently selected from hydrogen, fluorine, chlorine, cyano, methyl, ethyl, isopropyl, cyclopropyl, trifluoromethyl, isobutyl, R9 and R 13 and the carbon atoms to which they are attached together form a cyclopropyl group, and further, are selected from hydrogen, fluorine, deuterium, chlorine, methyl, hydroxyl, and amino. Specifically, X can be: Among them, X is All compounds of can be used as brain-penetrating BTK inhibitors or HER2 inhibitors, preferably, R9, R 13 All are selected from fluorine.

[0028] In some embodiments of the present application, the compound has a structure shown in Formula III, Formula IV, or a tautomer, mesomer, racemate, enantiomer, diastereomer or a mixture thereof, a pharmaceutically acceptable hydrate, solvate or salt thereof:

[0029]

[0030] Wherein, the structures of R1, R2, R3, R4, R6, and X are as described above; m, n, and n1 are also as described above; for example, X is wait.

[0031] In formula III-IV, n2 is selected from 0, 1, 2, 3, 4;

[0032] R8 is independently selected from hydrogen, halogen, hydroxyl, amino, cyano, alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl; further, R8 is independently selected from hydrogen, fluorine, chlorine, bromine, cyano, amino, C1-C3 alkyl, C1-C3 alkoxy, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl; further, the substituent is independently selected from hydrogen, fluorine, chlorine, bromine, cyano, trifluoromethyl, trifluoromethoxy, difluoromethoxy, methoxy, deuterated methoxy, cyclopropyl, cyclopropylmethoxy, ethyl, isopropyl, isobutyl; wherein the number of the substituents is an integer between 0 and 5 (including the endpoints); multiple substituents may be the same or different; in formula IV, the connection position of the substituted or unsubstituted pyridyl is not limited, and may be connected at the ortho position of N.

[0033] In some embodiments of the present application, the N-containing fused ring in Formula II-Formula IV can be used Instead, the single bonds at both ends are connecting bonds. In addition, in the X structures of formula II to formula IV, the single bonds with curved lines represent connecting bonds. The position of R6 is not limited; n1 is preferably 0, 1 or 2. When n=0, it is a five-membered ring; when n is 1, it is a six-membered ring, and so on.

[0034] Preferably, in formula II to formula IV, R1 is amino, R2 is hydrogen or chlorine, and R6 is hydrogen or monosubstituted fluorine; in formula II, R7 is substituted or unsubstituted phenyl or pyridyl; X is mainly an ether or amide structure and the nitrogen of the amide is connected to R7. Preferably, n is 0 or 1, m is 0 or 2, and R3 and R4 are both hydrogen, methyl, or form a cyclopropyl group with the carbon atom to which they are connected.

[0035] Specifically, the compound structure described in the present application is selected from one of the following (wherein, one end of the single bond is a methyl group, as shown in Formula 5 of Compound 5); further preferably, the compounds shown in Formula 2, Formula 5, Formula 34, Formula 42, Formula 89, Formula 100, Formula 101, Formula 103, Formula 106, Formula 109, Formula 111, Formula 114, Formula 116, Formula 118, Formula 121, Formula 125, Formula 130, Formula 145, Formula 146, Formula 152, and Formula 155 have better performance:

[0036]

[0037]

[0038]

[0039]

[0040]

[0041]

[0042]

[0043]

[0044]

[0045]

[0046]

[0047]

[0048] The present invention provides a pharmaceutical composition, wherein the active ingredient of the pharmaceutical composition is selected from one or a combination of two or more of the above-mentioned compounds or their stereoisomers, solvates, hydrates, pharmaceutically acceptable salts or cocrystals. In addition, the present invention has no special restrictions on the preparation type of the pharmaceutical composition.

[0049] The present invention provides the use of the above-mentioned compound or its stereoisomer, solvate, hydrate, pharmaceutically acceptable salt or cocrystal in the preparation of a protein kinase inhibitor; further, the kinase inhibitor is a BTK inhibitor or a HER2 inhibitor. Alternatively, the present invention provides the use of the above-mentioned compound or its stereoisomer, solvate, hydrate, pharmaceutically acceptable salt or cocrystal in the preparation of a drug for treating a disease caused by overexpression of BTK kinase or HER2 kinase.

[0050] The present invention provides use of the above-mentioned compound or its stereoisomers, solvates, hydrates, pharmaceutically acceptable salts or co-crystals in the preparation of a medicament for treating any one or more of autoimmune diseases, inflammatory diseases, thromboembolic diseases, allergies, infectious diseases, proliferative disorders and cancer.

[0051] Further, the disease can be selected from: arthritis, rheumatoid arthritis, urticaria, vitiligo, organ transplant rejection, ulcerative colitis, Crohn's disease, dermatitis, asthma, Sjögren's syndrome, systemic lupus erythematosus, multiple sclerosis, idiopathic thrombocytopenic purpura, rash, anti-neutrophil cytoplasmic antibody vasculitis, pemphigus vulgaris, chronic obstructive pulmonary disease, psoriasis; breast cancer, mantle cell lymphoma, ovarian cancer, esophageal cancer, laryngeal cancer, glioblastoma, neuroblastoma, gastric cancer, hepatocellular carcinoma, gastric cancer, glioma, endometrial cancer, melanoma, renal cancer, bladder cancer, melanoma, bladder cancer, Biliary tract cancer, kidney cancer, pancreatic cancer, lymphoma, hairy cell cancer, nasopharyngeal cancer, pharyngeal cancer, colorectal cancer, rectal cancer, brain and central nervous system cancer, cervical cancer, prostate cancer, testicular cancer, genitourinary tract cancer, lung cancer, non-small cell lung cancer, small cell cancer, lung adenocarcinoma, bone cancer, colon cancer, adenoma, pancreatic cancer, adenocarcinoma, thyroid cancer, follicular carcinoma, Hodgkin's leukemia, bronchial cancer, thyroid cancer, uterine body cancer, cervical cancer, multiple myeloma, acute myeloid leukemia, chronic myeloid leukemia, lymphocytic leukemia, chronic lymphoid leukemia, myeloid leukemia, non-Hodgkin's lymphoma, primary macroglobulinemia.

[0052] In the prior art, the inhibitory activity of ARQ-531 still needs to be improved. Its inhibitory activity against TMD8, REC-1 and other cells is poor, which leads to problems such as excessive clinical dosage and high side effects. In addition, ARQ-531 has poor selectivity and has high inhibitory activity against TEC and EGFR, which easily leads to side effects such as bleeding, diarrhea, and eczema. Moreover, its pharmacokinetics is not ideal. Preclinical studies have shown that in canine PK experiments, the bioavailability is only 38%. That is, ARQ-531 has a lot of room for improvement in terms of inhibitory activity, selectivity, and pharmacokinetics.

[0053] In the in vitro BTK inhibition and HER2 inhibition kinase activity test of the embodiment of the present invention, the compound powder can be dissolved in 100% DMSO to prepare a 10mM storage solution; -20 degrees dark frozen. During the kinase reaction, the test compound was tested at a concentration of 1μM, diluted to a 100% DMSO solution with a final concentration of 100 times in the 384source plate, 3 times the compound, 10 concentrations. In addition, the embodiment of the present invention also used the compound to conduct experiments such as liver microsome stability, rat PK, rat brain penetration rate, and pharmacodynamic model. Compared with the existing clinical drug (ARQ-531), the compound of the present invention as a BTK protein kinase inhibitor has advantages in BTK, BTK (C481S) inhibitory activity, liver microsome stability, rat pharmacokinetics, and toxicity. Compared with the existing marketed drug Tirabrutinib, the compound of the present invention as a BTK protein kinase inhibitor has advantages in BTK, BTK (C481S) inhibitory activity, cell activity, liver microsome stability, rat pharmacokinetics, and rat blood-brain barrier permeability.

[0054] The embodiments of the present invention design and synthesize multiple target compounds. The specific preparation process can be shown in the following formula. Intermediate A (also known as boronic acid or boronic ester compound shown in formula A) and intermediate B (bromo compound shown in formula B) are subjected to Suzuki reaction to synthesize intermediate C (intermediate shown in formula C), and then deprotected to obtain a compound with a structure shown in formula II. In a specific embodiment, intermediate C is prepared by coupling commercially available boronic acid A or homemade boronic ester A with homemade bromo compound B under palladium catalysis, and intermediate C is deprotected to obtain the embodiment compound. Compared with the clinical phase II drug ARQ-531, the compound of the present invention has significant improvements in the inhibitory activity of BTK and BTK (C481S), liver microsomal stability, and rat pharmacokinetics.

[0055] In addition, the following synthesis method of the embodiment of the present invention is simple and has a high yield.

[0056]

[0057] The present invention provides an intermediate compound for preparing the BTK inhibitor described above, which has the following structure:

[0058] R1, R2, R3, R4, m, and n are as described above; for example:

[0059] In addition, other intermediate compounds include wait. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] Figure 1The results of the drug efficacy model test of some compounds TMD8 of the present invention;

[0061] Figure 2 The results of the drug efficacy model test of some compounds TMD8 of the present invention;

[0062] Figure 3 The results of the drug efficacy test of some compounds of the present invention, DOHH-2-Luc, on the brain tumor model;

[0063] Figure 4 These are fluorescent photos of the test results of the efficacy of some compounds of the present invention, DOHH-2-Luc, in a brain tumor model. DETAILED DESCRIPTION

[0064] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0065] In order to further understand the present application, the compounds that can be used as BTK protein kinase inhibitors provided in the present application, as well as their preparation methods and uses are specifically described below in conjunction with the examples.

[0066] In the embodiments of the present invention, the structure of the compound is determined by mass spectrometry (MS) or nuclear magnetic resonance ( 1 H NMR) equipment. The term "room temperature" means between 10°C and 25°C. The chemical abbreviations have the following meanings:

[0067] DMF: N,N-dimethylformamide; DIEA: N,N-diisopropylethylamine;

[0068] HATU: O-(7-azabenzotriazol-1-yl)-N,N,N'-tetramethyluronium hexafluorophosphate;

[0069] PdCl2(dppf): [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride;

[0070] DCM: dichloromethane; TEA: triethylamine; TBDPSCl: lithium bistrimethylsilylamide;

[0071] 9-BBN: 9-boranebicyclo[3.3.1]nonane; Dess-Martin: Dess-Martin periodinane;

[0072] DME: ethylene glycol dimethyl ether; TosMIC: p-toluenesulfonylmethyl isocyanide;

[0073] t-BuOK: potassium tert-butoxide; Dibal-H: diisobutylaluminum hydride; THF: tetrahydrofuran;

[0074] NBS: N-bromosuccinimide; TBAF: tetrabutylammonium fluoride; DMSO: dimethyl sulfoxide;

[0075] LDA: lithium diisopropylamide;

[0076] HBTU: benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate;

[0077] NMP: N-methylpyrrolidone; BAST: bis(2-methoxyethyl)aminosulfur trifluoride;

[0078] PMDTA: pentamethyldiethylenetriamine; DMA: N,N-dimethylacetamide;

[0079] dppf: 1,1'-bis(diphenylphosphino)ferrocene; Pd2(dba)3: tris(dibenzylideneacetone)dipalladium;

[0080] TsCl: 4-toluenesulfonyl chloride; DMAP: 4-dimethylaminopyridine; PDC: pyridinium dichromate;

[0081] DIAD: diisopropyl azodicarboxylate; NCS: N-chlorosuccinimide.

[0082] Preparation of intermediate A-1:

[0083]

[0084] Compound A-1-1 (5.0 g, 53.1 mmol), DMF (50 mL), A-1-2 (11.6 g, 53.1 mmol) and DIEA (20.6 g, 159.3 mmol) were added to the reaction flask, the reaction liquid was replaced with nitrogen, cooled to 0 ° C, HATU (24.2 g, 63.7 mmol) was added in batches, the reaction mixture was slowly warmed to room temperature and stirred overnight, TLC showed that the raw materials were completely reacted. Water was added to the reaction system, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous Na2SO4, evaporated in vacuo, and purified by silica gel column to obtain 11.9 g product A-1-3, yield: 76%.

[0085] Compound A-1-3 (5.0 g, 16.9 mmol), dioxane (50 mL), bis-pinacol borate (5.2 g, 20.3 mmol) and potassium acetate (2.5 g, 25.4 mmol) were added to the reaction bottle, the reaction solution was replaced with nitrogen, PdCl2 (dppf) (500 mg, 0.68 mmol) was added to the reaction solution, the reaction solution was replaced with nitrogen again, the reaction mixture was heated to 90 ° C and stirred overnight, TLC showed that the raw materials reacted completely. After the reaction system was cooled, silica gel was directly added to mix the sample, and then the crude product was purified by silica gel column, and the crude product was slurried with petroleum ether to obtain 3.4 g product A-1, yield: 62%.

[0086] Preparation of intermediate A-2:

[0087]

[0088] Add compound A-2-1 (2.0 g, 11.8 mmol) and dioxane (20 mL) to the reaction bottle and cool in an ice-water bath. Add hydrogen peroxide (20 mL, 30%) dropwise to the reaction solution, then stir the reaction solution at room temperature overnight to stop the reaction. Add water to the reaction system, extract with ethyl acetate 4 times, combine the organic phases, wash with water, wash with saturated brine, evaporate in vacuo, and purify by silica gel column to obtain 1.6 g product A-2-2, yield: 96%.

[0089] Compound A-2-2 (1.00 g, 7.04 mmol), DMF (20 mL), p-bromoiodobenzene (1.99 g, 7.04 mmol), tetrabutylammonium bromide (230 mg, 0.704 mmol), potassium phosphate (2.99 g, 14.1 mmol) and cuprous iodide (140 mg, 0.704 mmol) were added to the reaction bottle, the reaction liquid was replaced with nitrogen, and then heated to 140 ° C. and stirred to react overnight. The reaction system was cooled to room temperature, water was added, and extracted with ethyl acetate 3 times. The organic phases were combined, washed with water, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 800 mg of product A-2-3, yield: 38%.

[0090] Compound A-2-3 (800 mg, 2.69 mmol), dioxane (16 mL), bis-pinacol borate (821 mg, 3.23 mmol) and potassium acetate (528 mg, 5.38 mmol) were added to the reaction bottle, the reaction solution was replaced with nitrogen, PdCl2 (dppf) (80 mg, 0.109 mmol) was added to the reaction solution, the reaction solution was replaced with nitrogen again, and the reaction mixture was heated to 80 ° C and stirred for 16 hours. After the reaction system was cooled, silica gel was directly added to mix the sample, and then purified by silica gel column to obtain 520 mg product A-2, yield: 56%.

[0091] Preparation of intermediate A-3:

[0092]

[0093] Compound A-3-1 (1.28 g, 10.5 mmol), DCM (40 mL), A-3-2 (1.0 g, 5.2 mmol), Cu(OAc)2 (945 mg, 5.2 mmol), TEA (1.58 g, 15.6 mmol) and 4A molecular sieve (1.66 g) were added to the reaction bottle, and the reaction solution was stirred at room temperature overnight. The reaction solution was filtered, silica gel was added to the filtrate, and then purified by silica gel column to obtain 600 mg of product A-3-3, yield: 43%.

[0094] The synthesis method of A-3 refers to the method for synthesizing A-2 from A-2-3.

[0095] Preparation of intermediate A-4:

[0096]

[0097] Compound A-4-1 (1.06 g, 6.3 mmol), DMF (10 mL), A-3-2 (1.0 g, 5.2 mmol) and potassium carbonate (1.45 g, 10.5 mmol) were added to the reaction flask, and the reaction solution was heated to 100 ° C and stirred for overnight reaction. TLC showed that the raw materials were completely reacted. Water was added to the reaction system, and the mixture was extracted twice with ethyl acetate. The organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous Na2SO4, and evaporated in vacuo to obtain 1.8 g of product A-4-2, with a yield of 100%. The product was used directly in the next step without purification.

[0098] The synthesis method of A-4 refers to the method for synthesizing A-2 from A-2-3.

[0099] The following compounds were synthesized by the method for preparing the above intermediates using commercially available corresponding raw materials.

[0100] Table 1: Structures and syntheses of intermediates A-5 to A-12

[0101]

[0102]

[0103] Preparation of intermediate A-13:

[0104]

[0105] Compound A-13-1 (1.00 g, 5.55 mmol), THF (15 mL) and triisopropyl borate (1.25 g, 6.66 mmol) were added to the reaction bottle, the reaction solution was cooled to -70 ° C, and LDA (2M, 3.3 mL, 6.6 mmol) was added dropwise to the reaction solution. After the addition, the reaction solution was slowly warmed to room temperature, dilute hydrochloric acid was added to quench the reaction, and the mixture was extracted twice with ethyl acetate. The organic phases were combined, washed with saturated brine, and dried by spin drying. The residue was purified by silica gel column to obtain 838 mg of product A-13-2, with a yield of 67%.

[0106] The synthesis method of A-13 refers to the method for synthesizing A-2 from A-2-1.

[0107] Preparation of intermediate A-14:

[0108]

[0109] Compound A-14-1 (500 mg, 3.90 mmol), acetonitrile (5 mL), potassium carbonate (647 mg, 4.68 mmol) and deuterated iodomethane (566 mg, 3.90 mmol) were added to the reaction bottle, and the reaction solution was heated to 50 ° C and stirred overnight. The reaction solution was poured into water, acidified with dilute hydrochloric acid, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, and dried, and the residue was purified by silica gel column to obtain 432 mg of product A-14-2, yield: 76%.

[0110] The synthesis method of A-14 refers to the method for synthesizing A-2 from A-2-2.

[0111] Preparation of intermediate A-15:

[0112]

[0113] Compound A-2-3 (500 mg, 1.68 mmol) and dichloromethane (8 mL) were added to the reaction flask, the reaction solution was cooled to -70 ° C, and a dichloromethane solution of boron tribromide (1 M, 5 mL, 5.0 mmol) was added dropwise to the reaction solution. After the addition was completed, the reaction was kept warm for 2 hours. The reaction solution was quenched with water, extracted twice with dichloromethane, the organic phases were combined, dried by spin drying, and the residue was purified by silica gel column to obtain 390 mg of product A-15-1, with a yield of 82%.

[0114] Compound A-15-1 (200 mg, 0.706 mmol), DMF (2 mL), cyclopropane bromide (171 mg, 1.41 mmol), cesium carbonate (276 mg, 0.847 mmol) and sodium iodide (53 mg, 0.353 mmol) were added to the reaction bottle, and the reaction solution was heated to 150 ° C for 15 hours. The reaction solution was poured into water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, and dried, and the residue was purified by silica gel column to obtain 121 mg product A-15-2, yield: 53%.

[0115] The synthesis method of A-15 refers to the method for synthesizing A-2 from A-2-3.

[0116] Preparation of intermediate A-16:

[0117]

[0118] The synthesis method of A-16 refers to the method of synthesizing A-13 from A-13-1.

[0119] Preparation of intermediate A-17:

[0120]

[0121] Compound A-17-1 (250 mg, 1.76 mmol), dichloromethane (5 mL), p-bromophenylboric acid (706 mg, 3.52 mmol), copper acetate (320 mg, 1.76 mmol), pyridine (418 mg, 5.28 mmol) and 4A molecular sieve (powder, 500 mg) were added to the reaction bottle. The reaction solution was stirred at room temperature for two days under atmospheric environment. Silica gel was directly added to the reaction solution and the sample was spin-dried and purified by silica gel column to obtain 367 mg of product A-17-2, with a yield of 70%.

[0122] The synthesis method of A-17 refers to the method for synthesizing A-2 from A-2-3.

[0123] Preparation of intermediate A-18:

[0124]

[0125] Add compound A-18-1 (500 mg, 2.44 mmol), toluene (10 mL), cyclopropylboric acid (315 mg, 3.66 mmol), potassium phosphate (1036 mg, 4.88 mmol), tricyclohexylphosphine (68 mg, 0.244 mmol), palladium acetate (30 mg) and water (0.5 mL) to the reaction bottle. Replace the reaction liquid with nitrogen and heat it to 100 ° C to react overnight. After the reaction liquid is cooled, add silica gel directly to the sample and spin dry. Purify it on a silica gel column to obtain 316 mg of product A-18-2, with a yield of 78%.

[0126] The synthesis method of A-18-3 refers to the method for synthesizing A-15-1 from A-2-3.

[0127] The synthesis method of A-18 refers to the method for synthesizing A-2 from A-2-2.

[0128] The following compounds were synthesized by the method for preparing the above intermediates using commercially available corresponding raw materials.

[0129] Table 2: Structure and synthesis of intermediates A-19 and A-20

[0130]

[0131] Preparation of intermediate A-21:

[0132]

[0133] Compound A-21-1 (500 mg, 1.91 mmol) and tetrahydrofuran (8 mL) were added to the reaction bottle, the reaction solution was replaced with nitrogen, cooled in an ice-salt bath, and methylmagnesium bromide (1M tetrahydrofuran solution, 2.3 mL, 2.3 mmol) was added dropwise to the reaction solution, and the mixture was heated to room temperature for 1 hour. The reaction solution was quenched with saturated aqueous ammonium chloride solution, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered to obtain 535 mg of product A-21-1, with a yield of 100%. The product was used directly in the next step without further purification.

[0134] The synthesis method of A-21 refers to the method for synthesizing A-2 from A-2-3.

[0135] Preparation of intermediate A-22:

[0136]

[0137] The synthesis of A-2 refers to Journal of Medicinal Chemistry, 2020, vol. 63, #10, 5102-5118.

[0138] The following compounds were synthesized by the method for preparing the above intermediates using commercially available corresponding raw materials.

[0139] Table 3: Structures and syntheses of intermediates A-23 to A-25

[0140]

[0141]

[0142] Preparation of intermediate A-26:

[0143]

[0144] The synthesis method of A-26-2 refers to the method of synthesizing A-18-2 from A-18-1.

[0145] Compound A-26-2 (500 mg, 2.57 mmol), methanol (8 mL) and aqueous sodium hydroxide solution (1 M, 5.1 mL, 5.1 mmol) were added to the reaction flask, and the reaction solution was reacted at room temperature overnight. The reaction solution was diluted with water, acidified with dilute hydrochloric acid, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and dried to obtain 440 mg of product A-26-3, yield: 95%. The product was used directly in the next step without further purification.

[0146] Compound A-26-3 (200 mg, 1.11 mmol), DMF (2 mL), 4-aminophenylboronic acid pinacol ester (268 mg, 1.22 mmol) and DIEA (430 mg, 3.33 mmol) were added to the reaction bottle. HATU (633 mg, 1.67 mmol) was added to the reaction solution at one time, and the mixture was reacted at room temperature overnight. The reaction solution was quenched with water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous Na2SO4, evaporated in vacuo, and purified by silica gel column to obtain 219 mg product A-26, yield: 52%.

[0147] Preparation of intermediate A-27:

[0148]

[0149] Compound A-21-1 (1000 mg, 3.83 mmol), S-tert-butylsulfenamide (511 mg, 4.21 mmol) and 1,4-dioxane (10 mL) were added to the reaction bottle, the reaction liquid was replaced with nitrogen, and tetraethyl titanate (2184 mg, 9.58 mmol) was added to the reaction liquid. The reaction liquid was heated to 100 ° C and stirred for 5 hours. The reaction liquid was cooled, quenched with water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 882 mg of product A-27-1, yield: 63%.

[0150] Compound A-27-1 (882 mg, 2.42 mmol) and tetrahydrofuran (14 mL) were added to the reaction bottle, the reaction liquid was replaced with nitrogen, cooled in an ice-salt bath, methylmagnesium bromide (1M tetrahydrofuran solution, 2.9 mL, 2.9 mmol) was added dropwise to the reaction liquid, and the mixture was heated to room temperature for 1 hour. The reaction liquid was quenched with saturated aqueous ammonium chloride solution, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 630 mg of product A-27-2, with a yield of 68%.

[0151] Compound A-27-2 (630 mg, 1.66 mmol) and methanol (10 mL) were added to the reaction flask, and then a solution of hydrogen chloride in 1,4-dioxane (4 M, 6 mL) was added. The reaction solution was reacted at room temperature for 1 hour and then concentrated to dryness under reduced pressure. Water was added to the residue, the base was adjusted with aqueous sodium hydroxide solution, and the mixture was extracted twice with ethyl acetate. The organic phases were combined, washed with water, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 371 mg of product A-27-3, with a yield of 81%.

[0152] The synthesis method of A-27 refers to the method for synthesizing A-2 from A-2-3.

[0153] Preparation of intermediate A-28:

[0154]

[0155] The synthesis method of A-28 refers to the method for synthesizing A-2 from A-2-3.

[0156] Preparation of intermediate A-29:

[0157]

[0158] Compound A-29-1 (1000 mg, 5.17 mmol) and tetrahydrofuran (10 mL) were added to the reaction bottle, the reaction liquid was replaced with nitrogen, and cooled in an ice-water bath. Isopropylmagnesium chloride (1M tetrahydrofuran solution, 6.2 mL, 6.2 mmol) was added dropwise to the reaction solution. After the addition, the reaction solution was heated to room temperature and stirred for 1 hour, and a solution of tetrahydrofuran (4 mL) of p-fluorobenzaldehyde (770 mg, 6.20 mmol) was added dropwise to the reaction solution. After the addition, the reaction solution was stirred at room temperature for 1 hour, and the reaction solution was quenched with saturated aqueous ammonium chloride solution, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 612 mg product A-29-2, yield: 50%.

[0159] Compound A-29-2 (612 mg, 2.56 mmol), dichloromethane (12 mL) and Dess-Martin oxidant (1632 mg, 3.85 mmol) were added to the reaction bottle, and the reaction solution was reacted at room temperature for 1 hour. TLC showed that the reaction was complete. Silica gel was directly added to the reaction solution and the sample was spin-dried and purified on a silica gel column to obtain 495 mg of product A-29-3, with a yield of 82%.

[0160] The synthesis method of A-29 refers to the method for synthesizing A-27 from A-21-1.

[0161] Preparation of intermediate A-30:

[0162]

[0163] Compound A-30-1 (500 mg, 2.92 mmol), acetonitrile (5 mL), p-bromophenol (607 mg, 3.51 mmol) and potassium carbonate (485 mg, 3.51 mmol) were added to the reaction bottle, and the reaction solution was heated to 70 ° C and stirred overnight. The reaction solution was cooled, filtered, washed with ethyl acetate, and the filtrate was evaporated in vacuo and purified by silica gel column to obtain 742 mg of product A-30-2, yield: 97%.

[0164] The synthesis method of A-30 refers to the method for synthesizing A-2 from A-2-3.

[0165] Preparation of intermediate A-31:

[0166]

[0167] Compound A-31-1 (500 mg, 2.82 mmol), 5-bromo-2-chloropyrimidine (546 mg, 2.82 mmol) and N-methylpyrrolidone (5 mL) were added to the reaction bottle, and the reaction solution was heated to 150 ° C and stirred for 2 hours. The reaction solution was cooled, diluted with water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 580 mg of product A-31-2, yield: 62%.

[0168] The synthesis method of A-31 refers to the method for synthesizing A-2 from A-2-3.

[0169] The following compounds were synthesized by the method for preparing the above intermediates using commercially available corresponding raw materials.

[0170] Table 4: Structure and synthesis of intermediates A-32 to A-33

[0171]

[0172] Preparation of intermediate A-34:

[0173]

[0174] Compound A-34-1 (500 mg, 2.94 mmol), DMF (5 mL), dimethylhydroxylamine hydrochloride (344 mg, 3.53 mmol) and DIEA (1520 mg, 11.8 mmol) were added to the reaction flask. HBTU (1449 mg, 3.82 mmol) was added to the reaction solution at one time under stirring. The reaction solution was stirred at room temperature overnight, poured into water, extracted with ethyl acetate 4 times, the organic phases were combined, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 600 mg of product A-34-2, yield: 96%.

[0175] Compound p-bromoiodobenzene (876 mg, 3.10 mmol) and tetrahydrofuran (10 mL) were added to the reaction bottle, the reaction liquid was replaced with nitrogen, and the dry ice / ethanol bath was cooled to -70 ° C. n-Butyl lithium (2.5 M, 1.24 mL, 3.10 mmol) was added dropwise to the reaction solution, and after stirring for 30 minutes, a solution of A-34-2 (600 mg, 2.81 mmol) in tetrahydrofuran (3 mL) was added dropwise to the reaction solution. After the dropwise addition, the reaction solution was slowly warmed to room temperature and reacted for 1 hour. The reaction solution was quenched with water, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 360 mg product A-34-3, yield: 41%.

[0176] The synthesis method of A-34 refers to the method for synthesizing A-2 from A-2-3.

[0177] Preparation of intermediate A-35:

[0178]

[0179] The synthesis method of A-35-2 refers to the method of synthesizing A-34-3 from A-34-2.

[0180] The synthesis method of A-35-3 refers to the method of synthesizing A-29-3 from A-29-2.

[0181] The synthesis method of A-35 refers to the method for synthesizing A-2 from A-2-3.

[0182] The following compounds were synthesized by the method for preparing the above intermediates using commercially available corresponding raw materials.

[0183] Table 5: Structures and syntheses of intermediates A-36 to A-39

[0184]

[0185]

[0186] Preparation of intermediate A-40:

[0187]

[0188] Compound A-38 (100 mg, 0.309 mmol), DMF (1 mL), iodomethane (48 mg, 0.340 mmol) and potassium carbonate (51 mg, 0.371 mmol) were added to the reaction bottle. The reaction solution was heated to 80 ° C for 5 hours. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, evaporated in vacuo, and purified by preparing a silica gel plate to obtain 62 mg of product A-40, yield: 60%.

[0189] The following compounds were synthesized by the method for preparing the above intermediates using commercially available corresponding raw materials.

[0190] Table 6: Structures and syntheses of intermediates A-41 to A-47

[0191]

[0192]

[0193] Preparation of intermediate A-48:

[0194]

[0195] Compound A-15-1 (200 mg, 0.706 mmol), DMF (4 mL) and sodium difluorochloroacetate (215 mg, 1.41 mmol) were added to the reaction flask, the reaction liquid was replaced with nitrogen, and heated to 100 ° C for 6 hours. The reaction liquid was cooled, poured into water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 128 mg of product A-48-1, yield: 54%.

[0196] The synthesis method of A-48 refers to the method for synthesizing A-2 from A-2-3.

[0197] The following compounds were synthesized by the method for preparing the above intermediates using commercially available corresponding raw materials.

[0198] Table 7: Structures and syntheses of intermediates A-49 to A-53

[0199]

[0200] Preparation of intermediate A-54:

[0201]

[0202] Compound A-54-1 (500 mg, 3.89 mmol), 5-bromo-2-chloropyrimidine (752 mg, 3.89 mmol), DMF (5 mL) and potassium carbonate (645 mg, 4.67 mmol) were added to the reaction bottle, and the reaction solution was heated to 100 ° C for 4 hours. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 511 mg of product A-54-2, yield: 46%.

[0203] The synthesis method of A-54 refers to the method for synthesizing A-2 from A-2-3.

[0204] Preparation of intermediate A-55:

[0205]

[0206] Compound A-55-1 (1.00 g, 7.35 mmol), Pd / C (10%, 200 mg) and methanol (25 mL) were added to the reaction flask, the reaction solution was replaced with hydrogen, and then stirred overnight under hydrogen pressure (balloon). The reaction solution was filtered and the filtrate was directly dried to obtain 1.00 g of product A-55-2, yield: 99%. The product was used directly in the next step without purification.

[0207] Compound A-55-2 (800 mg, 5.79 mmol) and tetrahydrofuran (10 mL) were added to the reaction bottle, the reaction liquid was replaced with nitrogen, and the dry ice / ethanol bath was cooled to -70 ° C. n-Butyl lithium (2.5 M, 2.8 mL, 6.95 mmol) was added dropwise to the reaction solution, and after stirring for 30 minutes, a solution of trimethyl borate (723 mg, 6.95 mmol) in tetrahydrofuran (3 mL) was added dropwise to the reaction solution. After the dropwise addition, the reaction solution was slowly warmed to room temperature and reacted for 30 minutes. The reaction solution was quenched with dilute hydrochloric acid, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 430 mg of product A-55-3, yield: 41%.

[0208] The synthesis method of A-55 refers to the method of synthesizing A-2 from A-2-1.

[0209] Preparation of intermediate A-56:

[0210]

[0211] Compound A-56-1 (500 mg, 3.90 mmol), dibromomethane (1018 mg, 5.85 mmol), DMF (8 mL) and potassium carbonate (1348 mg, 9.75 mmol) were added to the reaction bottle, and the reaction solution was heated to 100 ° C for 4 hours. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 320 mg of product A-56-2, yield: 59%.

[0212] The synthesis method of A-56 refers to the method for synthesizing A-55 from A-55-2.

[0213] The following compounds were synthesized by the method for preparing the above intermediates using commercially available corresponding raw materials.

[0214] Table 8: Structures and syntheses of intermediates A-57 to A-59

[0215]

[0216] Preparation of intermediate A-60:

[0217]

[0218] Compound A-60-1 (500 mg, 3.31 mmol), p-bromophenol (685 mg, 3.96 mmol), NMP (10 mL) and cesium carbonate (3.20 g, 9.90 mmol) were added to the reaction bottle, and the reaction solution was heated to 80 ° C and reacted overnight. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 830 mg of product A-60-2, yield: 82%.

[0219] The synthesis method of A-60 refers to the method for synthesizing A-2 from A-2-3.

[0220] The following compounds were synthesized by the method for preparing the above intermediates using commercially available corresponding raw materials.

[0221] Table 9: Structures and syntheses of intermediates A-61 to A-63

[0222]

[0223] Preparation of intermediate A-64:

[0224]

[0225] Compound A-64-1 (1.192 g, 8.05 mmol), bromobenzene (10.1 g, 64.3 mmol) and aluminum chloride (2.15 g, 16.1 mmol) were added to the reaction flask, the reaction liquid was replaced with nitrogen, and heated to 90 ° C for 3 hours. The reaction liquid was cooled, poured into dilute hydrochloric acid, extracted 3 times with dichloromethane, and the organic phases were combined. The organic phase was extracted 3 times with sodium carbonate aqueous solution, the aqueous phase was adjusted to pH 3 with dilute hydrochloric acid, the precipitated solid was filtered, washed with water, the filter cake was collected, and vacuum dried to obtain 2.0 g of product A-64-2, yield: 81%. The product was used directly in the next step without further purification.

[0226] The synthesis method of A-64-3 refers to the method for synthesizing A-2 from A-2-3.

[0227] Compound A-64-3 (200 mg, 0.57 mmol), DMF (3 mL), ammonium chloride (152 mg, 2.85 mmol) and DIEA (220 mg, 1.71 mmol) were added to the reaction flask. HBTU (324 mg, 0.85 mmol) was added to the reaction solution at one time under stirring. The reaction solution was stirred at room temperature overnight, poured into water, extracted with ethyl acetate three times, the organic phases were combined, washed with saturated brine, evaporated in vacuo, and purified by silica gel preparation plate to obtain 70 mg of product A-64, yield: 35%.

[0228] The following compounds were synthesized by the method for preparing the above intermediates using commercially available corresponding raw materials.

[0229] Table 10: Structures and syntheses of intermediates A-65 to A-67

[0230]

[0231] Preparation of intermediate A-68:

[0232]

[0233] Compound A-68-1 (200 mg, 1.39 mmol), p-bromophenol (361 mg, 2.09 mmol), NMP (2 mL) and potassium carbonate (384 mg, 2.78 mmol) were added to the reaction bottle, and the reaction solution was heated to 180 ° C for 8 hours. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, evaporated in vacuo, and purified by silica gel preparation plate to obtain 60 mg product A-68-2, yield: 15%.

[0234] The synthesis method of A-68 refers to the method for synthesizing A-2 from A-2-3.

[0235] Preparation of intermediate A-69:

[0236]

[0237] The synthesis method of A-69 refers to the method for synthesizing A-1 from A-1-1 and A-1-2.

[0238] Preparation of intermediate A-70:

[0239]

[0240] Compound A-21-1 (200 mg, 0.766 mmol), triethylsilane (267 mg, 2.31 mmol), dichloromethane (4 mL) and trifluoromethanesulfonic acid (35 mg, 0.231 mmol) were added to the reaction bottle, and the reaction solution was stirred at room temperature overnight. The reaction solution was poured into water, extracted twice with ethyl acetate, the organic phases were combined, evaporated in vacuo, and purified by silica gel column to obtain 190 mg of product A-70-1, yield: 100%.

[0241] The synthesis method of A-70 refers to the method for synthesizing A-2 from A-2-3.

[0242] Preparation of intermediate A-71:

[0243]

[0244] The synthesis method of A-71 refers to the method of synthesizing A-54 from A-54-1.

[0245] Preparation of intermediate A-72:

[0246]

[0247] Add compound A-72-2 (2.00 g, 8.06 mmol), DMF (15 mL), A-72-1 (1.31 g, 8.06 mmol), DIEA (3.12 g, 24.2 mmol) and HATU (4.60 g, 12.1 mmol) to the reaction bottle. Heat the reaction solution to 60 ° C and react overnight. The reaction solution is cooled, poured into water, extracted twice with ethyl acetate, the organic phases are combined, washed with water, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 1.56 g product A-72, yield: 49%.

[0248] Preparation of intermediate A-73:

[0249]

[0250] Compound A-21-1 (300 mg, 1.15 mmol) and BAST (3 mL) were added to the reaction tube, the reaction tube was sealed, and heated to 90 ° C for overnight reaction. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, evaporated in vacuo, and purified by silica gel preparation plate to obtain 270 mg of product A-73-1, yield: 83%.

[0251] The synthesis method of A-73 refers to the method for synthesizing A-2 from A-2-3.

[0252] Preparation of intermediate A-74:

[0253]

[0254] Compound A-74-1 (1.00 g, 7.93 mmol), PMDTA (1.44 g, 8.32 mmol) and tetrahydrofuran (10 mL) were added to the reaction flask, the reaction liquid was replaced with nitrogen, and the dry ice / ethanol bath was cooled to -70 ° C. n-Butyl lithium (2.5 M, 3.3 mL, 8.30 mmol) was added dropwise to the reaction solution, and after stirring for 2 hours, dry ice was carefully added to the reaction solution, and the reaction solution was slowly warmed to room temperature. The reaction solution was quenched with dilute hydrochloric acid, extracted with dichloromethane 3 times, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated in vacuo to obtain 1.00 g of product A-74-2, yield: 74%. The product was used directly in the next step without further purification.

[0255] Compound A-74-2 (800 mg, 4.70 mmol) and dichloromethane (8 mL) were added to the reaction flask, the reaction liquid was replaced with nitrogen, and cooled in an ice-water bath. A dichloromethane solution of boron tribromide (17%, 27.7 g, 18.8 mmol) was added dropwise to the reaction liquid. After the addition, the reaction liquid was heated to room temperature for 30 minutes, cooled in an ice-water bath again, and methanol was slowly added dropwise to quench the reaction. After the reaction liquid was directly evaporated in vacuum, it was purified by a silica gel column to obtain 560 mg of product A-74-3, with a yield of 76%.

[0256] Add compound A-74-3 (560 mg, 3.59 mmol), DMF (5 mL), methylamine alcohol solution (30%, 557 mg, 5.38 mmol), DIEA (1392 mg, 10.8 mmol) and HATU (1775 mg, 4.67 mmol) to the reaction bottle. Stir the reaction solution at room temperature overnight. Pour the reaction solution into water and extract with ethyl acetate 6 times. Combine the organic phases, evaporate in vacuo, and purify by silica gel column to obtain 254 mg product A-74-4, yield: 42%.

[0257] The synthesis method of A-74 refers to the method of synthesizing A-17 from A-17-1.

[0258] Preparation of intermediate A-75:

[0259]

[0260] The synthesis method of A-75-1 refers to the method of synthesizing A-34-2 from A-34-1.

[0261] Compound A-75-1 (500 mg, 1.91 mmol) and tetrahydrofuran (8 mL) were added to the reaction flask, the reaction solution was replaced with nitrogen, and cooled in an ice-salt bath. Phenylmagnesium bromide in tetrahydrofuran solution (1 M, 2.3 mL, 2.3 mmol) was added dropwise to the reaction solution, and the reaction solution was slowly heated to room temperature and stirred for 1 hour. The reaction solution was quenched with dilute hydrochloric acid, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated in vacuo to obtain 538 mg of product A-75-2, yield: 100%. The product was used directly in the next step without further purification.

[0262] The synthesis method of A-75 refers to the method of synthesizing A-73 from A-21-1.

[0263] Preparation of intermediate A-76:

[0264]

[0265] Compound A-76-1 (200 mg, 1.10 mmol), p-bromophenol (228 mg, 1.32 mmol), NMP (2 mL) and cesium carbonate (538 mg, 1.65 mmol) were added to the reaction bottle, and the reaction solution was heated to 80 ° C and reacted overnight. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 280 mg of product A-76-2, yield: 76%.

[0266] Compound A-76-2 (200 mg, 0.597 mmol), sodium methoxide (161 mg, 2.98 mmol) and NMP (2 mL) were added to the reaction flask, and the reaction solution was heated to 80 ° C and reacted overnight. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 116 mg of product A-76-3, yield: 56%.

[0267] The synthesis method of A-76 refers to the method for synthesizing A-2 from A-2-3.

[0268] Preparation of intermediate A-77:

[0269]

[0270] The synthesis method of A-77-3 refers to the method of synthesizing A-34-3 from A-34-1.

[0271] The synthesis method of A-77 refers to the method of synthesizing A-73 from A-21-1.

[0272] Preparation of intermediate A-78:

[0273]

[0274] The synthesis method of A-78-2 refers to the method of synthesizing A-34-2 from A-34-1.

[0275] Compound A-78-2 (474 ​​mg, 1.73 mmol), zinc cyanide (305 mg, 2.59 mmol), DMA (5 mL), zinc powder (47 mg), dppf (94 mg) and Pd2(dba)3 (94 mg) were added to the reaction bottle, the reaction liquid was replaced with nitrogen, heated to 110°C and reacted overnight. The reaction liquid was cooled, poured into water, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, evaporated in vacuo, and purified by silica gel column to obtain 473 mg of product A-78-3, yield: 100%.

[0276] The synthesis method of A-78-4 refers to the method of synthesizing A-34-3 from A-34-2.

[0277] The synthesis method of A-78 refers to the method for synthesizing A-73 from A-21-1.

[0278] The following compounds were synthesized by the method for preparing the above intermediates using commercially available corresponding raw materials.

[0279] Table 10: Structures and syntheses of intermediates A-79 to A-81

[0280]

[0281] Preparation of intermediate A-82:

[0282]

[0283] Add dichloromethane (5mL) to the reaction bottle, replace with nitrogen, cool to -40 ° C in a dry ice / acetonitrile bath, add titanium tetrachloride (1453mg, 7.66mmol), then slowly drop a toluene solution of dimethyl zinc (1M, 7.7mL, 7.7mmol), add, and keep warm for 30 minutes. Add a dichloromethane (3mL) solution of compound A-21-1 (500mg, 1.91mmol) to the reaction solution, add, keep warm for 1 hour, then slowly warm to room temperature and stir overnight. The reaction solution is quenched with water, extracted twice with dichloromethane, the organic phases are combined, washed with water, evaporated in vacuo, and purified by silica gel column to obtain 326mg product A-82-1, yield: 62%.

[0284] The synthesis method of A-82 refers to the method for synthesizing A-2 from A-2-3.

[0285] The following compounds were synthesized by the method for preparing the above intermediates using commercially available corresponding raw materials.

[0286] Table 12: Structure and synthesis of intermediates A-83 to A-100

[0287]

[0288]

[0289]

[0290] Preparation of intermediate A-101:

[0291]

[0292] Add p-bromoiodobenzene (1.71 g, 6.03 mmol) and tetrahydrofuran (15 mL) to the reaction bottle, replace with nitrogen, cool to -70 ° C in a dry ice / ethanol bath, then slowly drop n-butyl lithium (2.5 M, 2.4 mL, 6.03 mmol), add after addition, keep warm for 30 minutes. Weigh A-101-1 (1.00 g, 5.74 mmol) and dissolve in tetrahydrofuran (5 mL), add dropwise to the reaction solution, and after 10 minutes, slowly warm the reaction solution to room temperature. The reaction solution is quenched with water, extracted twice with ethyl acetate, the organic phases are combined, washed with water, evaporated in vacuo, and purified by silica gel column to obtain 1.4 g product A-101-2, yield: 73%.

[0293] The synthesis method of A-101 refers to the method for synthesizing A-2 from A-2-3.

[0294] Preparation of intermediate A-102:

[0295]

[0296] A-101-2 (550 mg, 1.66 mmol) and dichloromethane (6 mL) were added to the reaction bottle, replaced with nitrogen, and DAST (402 mg, 2.49 mmol) was added under ice bath, and the reaction was kept warm for 2 hours. The reaction solution was quenched with sodium bicarbonate aqueous solution, extracted twice with dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, evaporated in vacuo, and purified by silica gel preparation plate to obtain 410 mg product A-102, yield: 74%.

[0297] The synthesis method of A-102 refers to the method for synthesizing A-2 from A-2-3.

[0298] Preparation of intermediate A-103:

[0299]

[0300] A-103-1 (500 mg, 1.50 mmol) and aqueous hydrobromic acid solution (5 mL) were added to the reaction bottle, cooled in an ice bath, sodium nitrite (645 mg, 9.35 mmol) was added, and then the reaction was kept warm for 20 minutes. CuBr (2.69 g, 18.75 mmol) in aqueous hydrobromic acid solution (5 mL) was added to the reaction solution, and the reaction solution was slowly heated to room temperature for 3 hours. The reaction solution was diluted with water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, evaporated in vacuo, and purified by silica gel column to obtain 620 mg of product A-103-2, yield: 89%.

[0301] A-103-2 (200 mg, 0.43 mmol) and tetrahydrofuran (5 mL) were added to the reaction bottle, replaced with nitrogen, cooled to -70 ° C in a dry ice / ethanol bath, and then n-butyl lithium (2.5 M, 0.17 mL, 0.43 mmol) was slowly added dropwise. After the addition was completed, the reaction was kept warm for 1 hour. The reaction solution was quenched with dilute hydrochloric acid, extracted twice with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, and evaporated in vacuo to obtain 170 mg of crude A-103-3, which was used directly in the next step without purification.

[0302] The synthesis method of A-103 refers to the method for synthesizing A-2 from A-2-3.

[0303] Preparation of intermediate B-1:

[0304]

[0305] Compound B-1-1 (2.00 g, 17.5 mmol), imidazole (1.43 g, 21.0 mmol), DMF solution (10 mL) were added to the reaction bottle, the reaction solution was replaced with nitrogen, TBDPSCl (5.30 g, 19.3 mmol) was added in an ice-water bath, the mixture was added after the addition, ice was removed, and the reaction mixture was stirred at room temperature for 16 h. TLC showed that the reaction was complete. The reaction solution was poured into water, the reaction solution was extracted twice with ethyl acetate, combined, washed twice with water, then washed with saturated NaCl solution, and finally dried with anhydrous Na2SO4. After direct vacuum evaporation, 6.68 g of product B-1-2 was obtained, which was used directly in the next step without purification.

[0306] Add a tetrahydrofuran solution (35 mL) of B-1-2 (6.68 g, 18.9 mmol) to the reaction flask, replace the reaction liquid with nitrogen, and add 9-BBN (0.5 M, 91 mL) dropwise under an ice-water bath. After the addition is complete, the reaction mixture is stirred at room temperature for 16 h. TLC shows that the raw material has reacted completely. The reaction solution is cooled again with an ice-water bath, and 10% NaOH solution (24 mL) and 30% H2O2 solution (12 mL) are slowly added. Stirring is continued for 1 h. TLC shows that the intermediate has reacted completely. Pour the reaction solution into water, extract twice with ethyl acetate, combine, then wash twice with water, wash with saturated NaCl solution, directly add silica gel to mix the sample, and then purify by silica gel column to obtain 6.49 g of product B-1-3, two-step yield: 100%.

[0307] Add compound B-1-3 (6.49 g, 17.5 mmol) in dichloromethane (50 mL) to the reaction flask, replace the reaction liquid with nitrogen, add Dess-Martin oxidant (11.14 g, 26.3 mmol) to the reaction flask in an ice-water bath, stir for 1.5 h, TLC shows that the reaction is complete. Directly add silica gel to mix the sample, and then purify by silica gel column to obtain 6.45 g product B-1-4, yield: 100%.

[0308] Add compound B-1-4 (6.45 g, 17.5 mmol), ethanol (926 mg, 20.1 mmol), DME (60 mL) and TosMIC (3.92 g, 20.1 mmol) to the reaction bottle, replace with nitrogen, and cool in an ice-water bath. Add t-BuOK (3.83 g, 34.1 mmol) to the reaction solution, stir for 30 min, then slowly warm to room temperature, and continue stirring for 1.5 h. TLC shows that the raw materials have reacted completely. Pour the reaction solution into saturated NH4Cl solution (350 mL), extract twice with ethyl acetate, combine, add silica gel to mix, and then purify by silica gel column to obtain 2.27 g product B-1-5 (TLC shows two spots), yield: 34%.

[0309] Add a dichloromethane solution (35 mL) of B-1-5 (2.27 g, 5.97 mmol) to the reaction flask, replace the reaction liquid with nitrogen, cool it in a dry ice-ethanol bath, slowly drop Dibal-H (1 M, 9 mL) into the reaction liquid, stir at this temperature for 1.5 h, TLC shows that the reaction is complete, add the reaction liquid to cold dilute hydrochloric acid (1 M, 10 mL), stir until there are no bubbles, then add DCM to extract twice, combine the organic phases, wash with saturated NaCl solution, and finally dry with anhydrous Na2SO4. The reaction liquid is directly evaporated in vacuo to obtain 2.22 g of product B-1-6. Yield: 97%.

[0310] Compound B-1-6 (2.22 g, 5.81 mmol), THF (60 mL), water (30 mL) and K2CO3 (4.82 g, 34.8 mmol) were added to the reaction flask, and then KMnO4 (3.67 g, 23.2 mmol) was added in batches. The reaction solution was stirred at room temperature for 30 min. TLC showed that the raw materials reacted completely. Dilute hydrochloric acid was added to the reaction solution to adjust the acidity, and then NaHSO3 solution was added until the reaction solution was colorless. It was extracted twice with ethyl acetate, the organic phases were combined, washed with saturated NaCl solution, and finally dried with anhydrous Na2SO4. The reaction solution was directly evaporated in vacuum to obtain 1.89 g of product B-1-7. Yield: 82%.

[0311] Add compound B-1-7 (1.89 g, 4.74 mmol), 3-chloropyrazine-2-methylamine dihydrochloride (1.03 g, 4.74 mmol) and DMF (20 mL) to the reaction bottle, replace with nitrogen, cool in an ice-water bath, add HATU (2.16 g, 5.69 mmol) and DIEA (3.06 g, 23.7 mmol) to the reaction solution. Stir the reaction solution for 30 min and remove the ice, continue stirring at room temperature for 30 min. TLC shows that the raw material reaction is complete. Pour the reaction solution into water, extract twice with ethyl acetate, combine, then wash twice with water, then wash with saturated NaCl solution, add silica gel to mix the sample, and then purify by silica gel column to obtain 1.94 g product B-1-8 (TLC shows two spots). Yield: 78%.

[0312] Compound B-1-8 (790 mg, 1.51 mmol), DMF (0.8 mL) and ethyl acetate (8 mL) were added to the reaction flask, replaced with nitrogen, and phosphorus oxychloride (1.39 g, 9.08 mmol) was added dropwise to the reaction solution under an ice-water bath, and the reaction solution was stirred for 1 h. TLC showed that the raw material reaction was complete. The reaction solution was poured into a NaHCO3 solution (4.5 g NaHCO3 / 30 mL H2O) to quench, extracted twice with ethyl acetate, combined, then washed twice with water, then washed with a saturated NaCl solution, and finally dried with anhydrous Na2SO4. After direct vacuum evaporation, 680 mg of product B-1-9 was obtained (TLC showed two spots). Yield: 89.0%.

[0313] Add a DMF solution (7 mL) of B-1-9 (680 mg, 1.34 mmol) to the reaction flask, replace the reaction liquid with nitrogen, add NBS (287 mg, 1.61 mmol) under an ice-water bath, stir for 40 min, TLC shows that the reaction is complete. Pour the reaction liquid into water to quench, extract twice with ethyl acetate, combine, then wash twice with water, then wash with saturated NaCl solution, add silica gel to mix, and then purify by silica gel column to obtain 298 mg of product B-1-10-A (the point with less polarity on TLC, comes out first) and 339 mg of product B-1-10-B (the point with more polarity on TLC, comes out later). Total yield: 81%.

[0314] Compound B-1-10-A (298 mg, 0.509 mmol), ammonia water (6 mL), and n-butanol solution (3 mL) were added to the jar, the reaction bottle was sealed, heated to 95 ° C, and stirred for 16 h. The reaction solution was cooled, vacuum-dried, and then purified by silica gel column to obtain 184 mg of product B-1-A, with a yield of 64%.

[0315] B-1-B was prepared by reacting B-1-10-B with aqueous ammonia using the method for synthesizing B-1-A.

[0316] Preparation of intermediate B-2:

[0317]

[0318] Weigh NaH (8.56 g, 357 mmol) and suspend it in anhydrous THF (80 mL), heat it to 40-45 ° C, add the THF solution of compound B-2-1 (22.85 g, 149 mmol) dropwise, keep the temperature and stir to react for 15 minutes. Add the THF solution of ethyl acrylate dropwise, add it and continue to react for 15 minutes. After cooling the reaction solution to room temperature, add ice water, adjust the pH to 3 with concentrated HCl, extract twice with ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate under reduced pressure, and purify by column chromatography to obtain compound B-2-2 as a colorless oil (37.6 g, 83%).

[0319] Compound B-2-2 (37.6 g, 120 mmol) and sodium chloride (20.97 g, 359 mmol) were added to DMSO (170 mL) and H2O (5 mL) and reacted at 160°C for 1.7 h. The reaction solution was cooled, added to ice water, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound B-2-3 was purified by column chromatography as a colorless oil (21.6 g, 75%).

[0320] Weigh compound B-2-3 (21.6 g, 89.2 mmol), ethylene glycol (6.64 g, 107 mmol) and p-toluenesulfonic acid monohydrate (169 mg, 0.89 mmol) into toluene (180 mL) and reflux at 120° C. for 4 h. The reaction solution was cooled, added to a saturated aqueous NaHCO3 solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography to obtain compound B-2-4 as a light yellow liquid (23.7 g, 93%).

[0321] Weigh LAH (6.47 g, 166 mmol) in a three-necked flask, add anhydrous THF (150 mL), replace with nitrogen, and add a solution of compound B-2-4 (23.7 g, 82.8 mmol) in THF (100 mL) under ice-salt bath cooling. After the addition is complete, slowly warm to room temperature and react for 5 hours. Slowly add H2O / THF (1:1, 30 mL) to the reaction under an ice-water bath, then add 5N sodium hydroxide aqueous solution (8 mL), and stir at room temperature overnight. Add DCM / MeOH (5:1, 250 mL) to the reaction flask to dilute the reaction solution, filter, and rinse with DCM / MeOH (5:1). Add 50 g of silica gel to the filtrate, stir for 15 min, filter, and rinse. The filtrate is concentrated under reduced pressure to obtain compound B-2-5 (16.7 g, 99%).

[0322] Weigh compound B-2-5 (16.7 g, 82.6 mmol) into a reaction flask, add pyridine (100 mL), add TsCl (34.6 g, 182 mmol) under an ice-water bath, and stir at room temperature overnight. Dilute the reaction solution with ethyl acetate, wash with 10% citric acid solution and saturated sodium chloride, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The crude product was slurried with ethanol to obtain compound B-2-6 as a white solid (35 g, 83%).

[0323] Weigh compound B-2-6 (35 g, 68.5 mmol) into a reaction flask, add 1N HCl solution (260 mL) and THF (300 mL) and react at 80 ° C for 5 h. The reaction solution was extracted with ethyl acetate, the organic phase was washed with water, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Column chromatography was used to purify compound B-2-7 (26.2 g, 82%).

[0324] Add compound 1,3-dithiane (2.1 g, 17.4 mmol) and anhydrous THF (40 mL) to the reaction bottle, replace with nitrogen, add n-butyl lithium (2.5 M, 8.5 mL) dropwise under dry ice-ethanol bath cooling, and warm to 0 ° C for 1 h. After cooling in a dry ice-ethanol bath, add a THF solution of compound B-2-7 (6.5 g, 13.9 mmol) dropwise, and warm to room temperature for 1 h. The reaction solution was quenched by adding saturated NH4Cl solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by column chromatography gave compound B-2-8 (6.77 g, 83%).

[0325] Add compound B-2-8 (6.77 g, 11.5 mmol), NaOH (1.38 g, 34.6 mmol), THF (170 mL) to the reaction flask and reflux at 70 ° C overnight. Cool the reaction solution to room temperature, add water, extract with ethyl acetate, dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify by column chromatography to obtain compound B-2-9 (3.7 g, 77%).

[0326] Compound B-2-9 (3.7 g, 8.92 mmol), acetonitrile (50 mL) and water (12.5 mL) were added to the reaction bottle, and NBS (5.56 g, 31.2 mmol) was added under an ice-water bath. After the addition was completed, the mixture was moved to room temperature for 3 h. The reaction solution was added to a saturated NaHCO3 solution, extracted with ethyl acetate, the organic phases were combined, the organic phases were backwashed with a saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude compound B-2-10 without further purification.

[0327] The crude compound B-2-10 obtained above was dissolved in ethanol, and NaBH4 (508 mg, 13.4 mmol) was added under an ice-water bath, and the mixture was moved to room temperature for 1 h. The reaction solution was quenched by adding saturated NH4Cl solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound B-2-11 (2.66 g, two-step yield 91%) was obtained by column chromatography purification.

[0328] Add compound B-2-11 (2.56 g, 7.84 mmol), DMAP (287 mg, 2.35 mmol), imidazole (1.06 g, 15.7 mmol) and DMF (15 mL) to the reaction bottle, then add TBDPSCl (2.59 g, 9.41 mmol) and react at room temperature for 0.5 h. Add the reaction solution into water, extract with ethyl acetate, combine the organic phases, backwash the organic phases with saturated NaCl solution, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify by column chromatography to obtain compound B-2-12 (4.0 g, 90%).

[0329] Compound B-2-12 (4.0 g, 7.08 mmol) and methanol (120 mL) were added to the reaction flask, and Mg (1.89 g, 77.9 mmol) was added under stirring at room temperature. After 30 minutes, the reaction was violently exothermic, and the reaction was stirred overnight. The reaction solution was added to a saturated NH4Cl solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound B-2-13 (2.4 g, 83%) was obtained by column chromatography.

[0330] Add compound B-2-13 (2.4 g, 5.85 mmol) and DMF (30 mL) to the reaction flask, add PDC (6.6 g, 17.6 mmol) under ice-water bath, after addition, move to room temperature to react for 2 h. Add ethyl acetate to the reaction flask to dilute the reaction solution, extract with water, combine the organic phases, backwash the organic phases with saturated NaCl solution, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify by column chromatography to obtain compound B-2-14 (1.99 g, 80%).

[0331] Add compound B-2-14 (1.99 g, 4.69 mmol), 3-chloropyrazine-2-methylamine dihydrochloride (1.02 g, 4.69 mmol) and DMF (10 mL) to the reaction bottle, then add HBTU (2.13 g, 5.62 mmol) and DIEA (2.42 g, 18.8 mmol) to the reaction solution, and react at room temperature for 1 h. Add the reaction solution to water, extract twice with ethyl acetate, combine the organic phases, backwash with saturated NaCl solution, dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure. Purify by column chromatography to obtain compound B-2-15 (1.99 g, 77%).

[0332] Compound B-2-15 (1.59 g, 2.89 mmol) was added to the reaction bottle, and DCM (30 mL) was added to dissolve. Pyridine (1.83 g, 23.1 mmol) and trifluoromethanesulfonic anhydride (4.89 g, 17.3 mmol) were added under nitrogen protection in an ice-water bath. After addition, the mixture was moved to room temperature for 4 h. The reaction solution was added to a saturated NaHCO3 solution, extracted with ethyl acetate, the organic phases were combined, the organic phases were backwashed with a saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude compound B-2-16 without further purification.

[0333] The crude compound B-2-16 obtained above was dissolved in DMF (8 mL), and NBS (566 mg, 3.18 mmol) was added to react at room temperature for 0.5 h. The reaction solution was added to NaHCO3 solution, extracted with ethyl acetate, the organic phases were combined, backwashed with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Column chromatography was used to purify the compound B-2-17 (1.43 g, two-step yield 81%).

[0334] Compound B-2-17 (1.43 g, 2.34 mmol), aqueous ammonia (20 mL), and n-butanol (8 mL) were added to the jar, and the reaction system was heated to 95°C and stirred for 16 h. The reaction solution was vacuum dried and purified by column chromatography to obtain compound B-2 (1.2 g, 87%).

[0335] Preparation of intermediate B-3:

[0336]

[0337] B-3-1 was prepared according to the method in the literature (Angew. Chem. Int. Ed. 2020, 59, 7161-7167).

[0338] Compound B-3-1 (1.25 g, 6.71 mmol), imidazole (548 mg, 8.06 mmol) and DMF (12 mL) were added to the reaction flask, and TBDPSCl (1.94 g, 7.05 mmol) was added to react at room temperature overnight. The reaction solution was added to water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound B-3-2 (2.85 g, 100%). No further purification was performed.

[0339] Compound B-3-2 (2.85 g, 6.71 mmol) was dissolved in ethanol (25 mL), and an aqueous solution (10 mL) of sodium hydroxide (403 mg, 10.1 mmol) was added. The reaction solution was heated to 60 ° C and reacted overnight. The reaction solution was cooled, added to water, and the acid was adjusted with dilute hydrochloric acid. It was extracted twice with ethyl acetate, and the organic phases were combined, washed with water, washed with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound B-3-3 (2.53 g,

[0340] 95%) without further purification.

[0341] The method for preparing B-3 from B-3-3 refers to the method for preparing B-2 from B-2-14.

[0342] Preparation of intermediate B-4:

[0343]

[0344] Compound B-4-1 (100 mg, 0.383 mmol), B-1-3 (170 mg, 0.460 mmol), triphenylphosphine (251 mg, 0.958 mmol) and THF (2 mL) were added to the reaction flask, the reaction liquid was replaced with nitrogen, and heated to 60°C. DIAD (194 mg, 0.958 mmol) was added dropwise to the reaction liquid, and the reaction was kept warm overnight. The reaction liquid was cooled, concentrated to dryness under reduced pressure, and purified by silica gel column to obtain compounds B-4-A (52 mg, a point with low polarity) and B-4-B (140 mg, a point with high polarity, containing triphenylphosphine oxide as an impurity). Total yield: 82%.

[0345] Preparation of intermediate B-5:

[0346]

[0347] Compound B-1-3 (500 mg, 1.35 mmol), TEA (273 mg, 2.70 mmol), DCM (5 mL) and p-toluenesulfonyl chloride (309 mg, 1.62 mmol) were added to the reaction bottle, and the reaction solution was stirred at room temperature for 2 h. TLC showed almost no reaction. DMAP (198 mg, 1.62 mmol) was added to the reaction solution and the reaction was allowed to proceed overnight. The reaction solution was added to water, acidified with dilute hydrochloric acid, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated NaCl solution, concentrated under reduced pressure, and purified by silica gel column to obtain compound B-5-1 (550 mg, 78%).

[0348] Compound B-5-1 (200 mg, 0.381 mmol), 3-bromo-4-chloro-1H-pyrazolo[4,3-C]pyridine (89 mg, 0.381 mmol), cesium carbonate (149 mg, 0.457 mmol) and DMA (2 mL) were added to the reaction bottle, and the reaction solution was heated to 100 ° C and stirred overnight. The reaction solution was cooled, added to water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated NaCl solution, concentrated under reduced pressure, and purified on a silica gel preparation plate to obtain compound B-5-2 (60 mg, 27%).

[0349] Add compound B-5-2 (60 mg, 0.103 mmol), ammonia (2 mL), and n-butanol (1 mL) to the sealed tube, heat the reaction system to 100 ° C, and stir overnight. The reaction solution is cooled, vacuum-dried, and purified on a silica gel preparation plate to obtain compound B-5 (38 mg, 66%).

[0350] Preparation of intermediate B-6:

[0351]

[0352] Compound B-1-10-B (200 mg, 0.342 mmol), THF (3 mL) and TBAF (1 M, 0.7 mL, 0.7 mmol) were added to the reaction bottle, and the reaction solution was stirred at room temperature for 4 h. The reaction solution was directly purified on a silica gel preparation plate to obtain compound B-6-1 (108 mg, 91%).

[0353] Compound B-6-1 is oxidized with PDC (refer to the synthesis of B-2-14 from B-2-13) to obtain B-6-2.

[0354] Compound B-6-2 (90 mg, 0.25 mmol), potassium carbonate (69 mg, 0.50 mmol), DMF (1 mL) and iodomethane (53 mg, 0.374 mmol) were added to the reaction bottle, and the reaction solution was stirred at room temperature overnight. The reaction solution was added to water, extracted twice with ethyl acetate, the organic phases were combined, washed with water, washed with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified on a silica gel preparation plate to obtain compound B-6-3 (80 mg, 85%).

[0355] Compound B-6-3 (80 mg, 0.21 mmol) and THF (2 mL) were added to the reaction bottle, replaced with nitrogen, and cooled in an ice-water bath. Tetraisopropyl titanate (28 mg, 0.10 mmol) was added to the reaction solution, and then ethylmagnesium bromide (0.6 mL, 0.6 mmol, 1 M) was slowly added dropwise. After the dropwise addition, the reaction solution was warmed to room temperature and stirred overnight. The reaction solution was poured into an aqueous ammonium chloride solution for quenching, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, concentrated under reduced pressure, and purified on a silica gel preparation plate to obtain 22 mg of compound B-6-4, with a yield of 28%.

[0356] The method for preparing B-6 from B-6-6 refers to the method for preparing B-2 from B-2-17.

[0357] Preparation of intermediate B-7:

[0358]

[0359] Compound B-6-3 (100 mg, 0.267 mmol) and THF (2 mL) were added to the reaction flask, replaced with nitrogen, and cooled in an ice-water bath. Methylmagnesium bromide (0.8 mL, 0.8 mmol, 1 M) was added dropwise to the reaction solution. After the addition, the reaction solution was warmed to room temperature and stirred overnight. The reaction solution was poured into an aqueous ammonium chloride solution for quenching, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, concentrated to dryness under reduced pressure, and purified on a silica gel preparation plate to obtain 65 mg of compound B-7-1, with a yield of 65%.

[0360] The method for preparing B-7 from B-7-1 refers to the method for preparing B-2 from B-2-17.

[0361] Preparation of intermediate B-8:

[0362]

[0363] Add compound B-8-1 (CAS: 652-67-5, 1.00 g, 6.84 mmol), imidazole (559 mg, 8.21 mmol) and DMF (15 mL) to the reaction bottle, then add TBDPSCl (1.88 g, 6.84 mmol), and react at room temperature overnight. Pour the reaction solution into water, extract twice with ethyl acetate, combine the organic phases, wash with water, wash with saturated NaCl solution, concentrate under reduced pressure, and purify on a silica gel column to obtain compound B-8-2 (1.63 g, 62%).

[0364] The method for preparing B-8 from B-8-2 refers to the method for preparing B-1-A(B) from B-1-3.

[0365] Preparation of intermediate B-9:

[0366]

[0367] Compound B-9-1 (8.0 g, 76.1 mmol), dioxane (120 mL) and RaneyNi (about 1 g) were added to the reaction bottle, the reaction solution was replaced with hydrogen, heated to 90 ° C under hydrogen bag pressure, stirred and reacted overnight, TLC showed that the raw materials were basically reacted. The reaction solution was cooled, filtered, and the filtrate was decompressed and dried to obtain 8.3 g of product B-9-2, yield: 100%. The product was used directly in the next step without purification.

[0368] B-9-2 is condensed with B-1-7, ring-closed, and brominated to obtain B-9. The specific method refers to the method for preparing B-1-10-A(B) from B-1-7.

[0369] Preparation of intermediate B-10:

[0370]

[0371] Add compound B-1-9 (200 mg, 0.395 mmol) and tetrahydrofuran (3 mL) to the reaction bottle, replace the reaction liquid with nitrogen, cool to -70 ° C in a dry ice / ethanol bath, and drop n-butyl lithium (2.5 M, 0.19 mL, 0.474 mmol) into the reaction liquid. After the drop is complete, keep warm for 30 minutes. Add iodomethane (112 mg, 0.790 mmol) to the reaction liquid, and after the drop is complete, slowly warm the reaction liquid to room temperature. Add ammonium chloride aqueous solution to quench the reaction, extract twice with ethyl acetate, combine the organic phases, concentrate under reduced pressure, and purify by column chromatography to obtain B-10-1 (160 mg, 78%).

[0372] B-10-1 is brominated with NBS and then ammonolyzed to obtain B-10. The specific method refers to the method for preparing B-1-A(B) from B-1-9.

[0373] Example 1: Preparation of Compound 1

[0374]

[0375] Compound B-1-B (205 mg, 0.362 mmol), A-1 (161 mg, 0.471 mmol), Na2CO3 (77 mg, 0.724 mmol), PdCl2 (dppf) (20 mg), dioxane (6 mL) and water (2 mL) were added to the reaction bottle, replaced with nitrogen, heated to 95 ° C for 2.5 h, and TLC showed that the reaction was complete. The reaction solution was diluted with ethyl acetate, and silica gel was directly added to stir the sample, and then purified by silica gel column to obtain 182 mg of product C-1-B, yield: 72%.

[0376] Compound C-1-B (182 mg, 0.260 mmol) and tetrahydrofuran (4 mL) were added to the reaction flask, TBAF (1 M, 0.39 mL) was added to the reaction solution, and the reaction solution was stirred at room temperature for 1.5 h. TLC showed that the reaction was complete. The reaction solution was directly purified by preparative silica gel plate (DCM / MeOH=15 / 1) to obtain 70 mg of product 1-B, yield: 58%.

[0377] The product structure was characterized by nuclear magnetic resonance and mass spectrometry, and the results are as follows:

[0378] 1 H NMR(400MHz,d6-DMSO)δ1.56-1.60(1H,m),1.94-2.07(2H,m),2.20-2.25(1H,m),3.28-3.31(1H,m),3.38-3 .42(2H,m),3.47-3.51(1H,m),3.77(1H,dd,J=11.8Hz,3.2Hz),4.11(1H,dd,J=11.8Hz,1.8Hz),4.59(1H,t, J=5.8Hz),6.03(2H,brs),7.07(1H,d,J=5.0Hz),7.20(1H,ddd,J=7.4Hz,4.9Hz,1.0Hz),7.63(2H,dd,J=10. 4Hz, 5.4Hz), 7.85-7.90 (1H, m), 7.98-8.02 (2H, m), 8.21 (1H, d, J = 8.4Hz), 8.41-8.43 (1H, m), 10.97 (1H, s).

[0379] MS (ESI) m / z (M+H) + :463.0.

[0380] 1-A was prepared by using A-1 and B-1-A using the method for synthesizing 1-B.

[0381] The product structure was characterized by nuclear magnetic resonance and mass spectrometry, and the results are as follows:

[0382] 1H NMR(400MHz,d6-DMSO)δ1.38-1.50(1H,m),1.73-1.75(1H,m),1.80-1.92(1H,m),2.12-2.15(1H,m ),3.36-3.47(3H,m),3.62(1H,t,J=11.0Hz),4.07-4.10(1H,m),4.69(1H,t,J=5.5Hz),6.02(2H,s) ,7.07(1H,d,J=5.0Hz),7.20(1H,dd,J=6.9Hz,5.2Hz),7.61(1H,t,J=7.9Hz),7.76(1H,d,J=5.0Hz) ,7.83-7.91(1H,m),7.95-8.04(2H,m),8.21(1H,d,J=8.4Hz),8.42(1H,d,J=3.8Hz),10.97(1H,s).

[0383] MS (ESI) m / z (M+H) + :463.1.

[0384] Example 2: Preparation of Compound 2

[0385]

[0386] 2-A is prepared by reacting A-5 with B-1-A using the method for synthesizing 1-B.

[0387] The product structure was characterized by nuclear magnetic resonance and mass spectrometry, and the results are as follows:

[0388] 1 H NMR(400MHz,d6-DMSO)δ1.41-1.51(1H,m),1.50-1.78(1H,m),1.85-1.97(1H,m),2.13-2.15(1H,m),3.3 5-3.49(4H,m),3.65(1H,t,J=11.0Hz),4.10(1H,ddd,J=11.0Hz,3.6Hz,1.6Hz),4.69(1H,t,J=5.6Hz),6 .13(2H,brs),7.09(1H,d,J=4.9Hz),7.19(1H,dd,J=6.9Hz,5.2Hz),7.76(3H,dd,J=9.5Hz,6.7Hz),7.83 -7.90(1H,m),8.16(2H,d,J=8.4Hz),8.23(1H,d,J=8.4Hz),8.41(1H,dd,J=4.8Hz,1.0Hz),10.84(1H,s).

[0389] MS (ESI) m / z (M+H) + :445.2.

[0390] Compound 2-A was separated by SFC to give 2-A-P1 (first eluting peak) and 2-A-P2 (latter eluting peak).

[0391] 2-B is prepared by reacting A-5 with B-1-B using the method for synthesizing 1-B.

[0392] The product structure was characterized by nuclear magnetic resonance and mass spectrometry, and the results are as follows:

[0393] 1 H NMR(400MHz,d6-DMSO)δ1.58-1.63(1H,m),1.95-2.02(1H,m),2.08-2.17(1H,m),2.24-2.28(1H,m),3.39 -3.51(4H,m),3.78(1H,dd,J=11.7,3.2Hz),4.10(1H,d,J=10.1Hz),4.60(1H,t,J=5.2Hz),6.14(2H,brs) ,7.09(1H,d,J=4.9Hz),7.18(1H,dd,J=6.9Hz,5.3Hz),7.63(1H,d,J=5.0Hz),7.76(2H,d,J=8.3Hz),7.84 -7.88(1H,m),8.16(2H,d,J=8.3Hz),8.23(1H,d,J=8.3Hz),8.41(1H,dd,J=4.8Hz,1.0Hz),10.84(1H,s).

[0394] MS (ESI) m / z (M+H) + :445.2.

[0395] Compound 2-A was separated by SFC to give 2-A-P1 (first eluting peak) and 2-A-P2 (latter eluting peak).

[0396] Preparative SFC conditions:

[0397] Instrument:SFC-80(Thar,Waters)

[0398] Column:CHIRALCEL OJ(30*250mm5μm)(Daicel)

[0399] Column temperature:35℃

[0400] Mobile phase:A=CO2 Co-Solvent B=ETOH

[0401] Cycle Time:12.5min Run Time:21min

[0402]

[0403] 2-B is prepared by reacting A-5 with B-1-B using the method for synthesizing 1-B.

[0404] The product structure was characterized by nuclear magnetic resonance and mass spectrometry, and the results are as follows:

[0405] 1 H NMR(400MHz,d6-DMSO)δ1.58-1.63(1H,m),1.95-2.02(1H,m),2.08-2.17(1H,m),2.24-2.28(1H,m),3.39 -3.51(4H,m),3.78(1H,dd,J=11.7,3.2Hz),4.10(1H,d,J=10.1Hz),4.60(1H,t,J=5.2Hz),6.14(2H,brs) ,7.09(1H,d,J=4.9Hz),7.18(1H,dd,J=6.9Hz,5.3Hz),7.63(1H,d,J=5.0Hz),7.76(2H,d,J=8.3Hz),7.84 -7.88(1H,m),8.16(2H,d,J=8.3Hz),8.23(1H,d,J=8.3Hz),8.41(1H,dd,J=4.8Hz,1.0Hz),10.84(1H,s).

[0406] MS (ESI) m / z (M+H) + :445.2.

[0407] Example 3: Preparation of Compound 3

[0408]

[0409] Compound 2-B (50 mg, 0.113 mmol), NCS (16.5 mg, 0.124 mmol) and glacial acetic acid (1 mL) were added to the reaction bottle, and the reaction solution was heated to 80°C for 2 h. The reaction solution was concentrated under reduced pressure, and an aqueous sodium bicarbonate solution was added, extracted twice with ethyl acetate, the organic phases were combined, washed with a saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by a silica gel preparation plate to obtain 28 mg of product 3, yield: 52%.

[0410] MS (ESI) m / z (M+H) + :479.2.

[0411] Examples 4 to 119: Preparation of Compounds 4 to 119

[0412] Compounds 4 to 119 were prepared by using different intermediates using the method for preparing compound 1-B or 3. The intermediate numbers, structural formulas, MS and 1 The H-NMR data are shown in Table 13.

[0413] Table 13: Structure, MS and 1 H-NMR data

[0414]

[0415]

[0416]

[0417]

[0418]

[0419]

[0420]

[0421]

[0422]

[0423]

[0424]

[0425]

[0426]

[0427]

[0428]

[0429]

[0430]

[0431]

[0432]

[0433]

[0434]

[0435]

[0436] Example 120: Preparation of Compound 120

[0437]

[0438] Compound B-1-4 (3.10 g, 8.41 mmol), methanol (31 mL), hydroxylamine hydrochloride (1.17 g, 16.8 mmol) and sodium acetate (2.07 g, 25.2 mmol) were added to the reaction flask. The reaction solution was stirred at room temperature overnight. Poured into water, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, concentrated under reduced pressure, and then purified by silica gel column to obtain 1.90 g of product 120-1, yield: 59%.

[0439] Compound 120-1 (1.90 g, 4.95 mmol) and tetrahydrofuran (20 mL) were added to the reaction bottle, cooled in an ice bath, and lithium aluminum tetrahydride (376 mg, 9.91 mmol) was added to the reaction solution in batches. The reaction solution was warmed to room temperature and reacted for 3 h. The reaction solution was cooled in an ice bath again, and water (380 mg), 15% NaOH aqueous solution (380 mg), and water (1.14 g) were slowly added dropwise to quench the reaction. The resulting suspension was filtered, washed with DCM / MeOH (10 / 1), and the filtrate was concentrated under reduced pressure and then purified by silica gel column to obtain 200 mg of product 120-2, yield: 31%.

[0440] Compound 2,4-dichloro-3-nitropyridine (294 mg, 1.52 mmol), DMF (2 mL), 120-2 (200 mg, 1.52 mmol) and triethylamine (231 mg, 2.29 mmol) were added to the reaction flask. The reaction solution was stirred at room temperature for 4 h. The reaction solution was poured into water and extracted with ethyl acetate three times. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 464 mg of product 120-3, with a yield of 100%. The product was not further purified.

[0441] Compound 120-3 (464 mg, 1.61 mmol), isopropanol (5 mL), bis-(4-methoxybenzyl)-amine (415 mg, 1.61 mmol) and triethylamine (212 mg, 2.10 mmol) were added to the reaction bottle. The reaction solution was heated to 95 ° C and stirred for 4 h. The reaction solution was cooled, concentrated to dryness under reduced pressure, and then purified by silica gel column to obtain 540 mg of product 120-4, yield: 66%.

[0442] Compound 120-4 (388 mg, 0.76 mmol), DMF (4 mL), imidazole (78 mg, 1.14 mmol), DMAP (10 mg, 0.076 mmol) and tert-butyldiphenylsilyl chloride (210 mg, 0.76 mmol) were added to the reaction bottle. The reaction solution was heated to 60 ° C and stirred overnight. TLC showed that a lot of raw materials remained. Imidazole (150 mg), DMAP (40 mg) and tert-butyldiphenylsilyl chloride (100 mg) were added to the reaction solution, and the temperature was raised to 80 ° C for 2 h. Tert-butyldiphenylsilyl chloride (200 mg) was continued to be added to the reaction solution. After 2 h, TLC showed that the reaction was complete. The reaction solution was cooled, poured into water, extracted with ethyl acetate 3 times, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by silica gel column to obtain 650 mg of product 120-5, yield: 100%.

[0443] Compound 120-5 (650 mg, 0.87 mmol), methanol / glacial acetic acid (5 mL / 5 mL) and iron powder (486 mg, 8.7 mmol) were added to the reaction flask. The reaction solution was stirred at room temperature for 4 h. The reaction solution was slowly poured into a NaHCO3 aqueous solution, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 612 mg of product 120-6, yield: 98%. The product was not further purified.

[0444] Compound 120-6 (612 mg, 0.85 mmol), acetonitrile (6 mL) and N,N'-carbonyldiimidazole (280 mg, 1.71 mmol) were added to the reaction flask. The reaction solution was heated to 80°C and stirred overnight. The reaction solution was cooled, concentrated to dryness under reduced pressure, and then purified by silica gel column to obtain 470 mg of product 120-7, with a yield of 74%.

[0445] Compound 120-7 (470 mg, 0.63 mmol), dichloromethane (15 mL), 4-phenoxyphenylboronic acid (271 mg, 1.27 mmol), acetic acid ketone (115 mg, 0.63 mmol), 4A molecular sieves (500 mg) and triethylamine (192 mg, 1.90 mmol) were added to the reaction bottle. The reaction solution was stirred at room temperature for 36 h. The reaction solution was filtered through diatomaceous earth and washed with ethyl acetate. The filtrate was concentrated to dryness under reduced pressure and then purified by silica gel column to obtain 240 mg of product 120-8, yield: 41%.

[0446] Compound 120-8 (260 mg, 0.29 mmol), dichloromethane (4 mL) and trifluoroacetic acid (4 mL) were added to the reaction flask. The reaction solution was heated to 50 ° C and stirred for 3 h. The reaction solution was cooled, concentrated to dryness under reduced pressure, and an aqueous NaHCO3 solution was added. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was dissolved in tetrahydrofuran (2 mL), TBAF (1 M, 0.2 mL) was added, and stirred at room temperature for 1 h. The reaction solution was directly purified on a silica gel preparation plate to obtain 40 mg of product 120, with a yield of 30%.

[0447] The product structure was characterized by nuclear magnetic resonance and mass spectrometry, and the results are as follows:

[0448] 1 H NMR(400MHz,d6-DMSO)δ1.37-1.53(1H,m),1.56-1.71(1H,m),1.81-1.95(1H,m),2.12-2 .29(1H,m),2.34-2.44(4H,m),3.40-3.54(1.5H,m),3.59-3.70(1H,m),3.77-4.02(1H,m ),4.17-4.39(1H,m),4.71(0.5H,t,J=5.7Hz),4.76-4.83(2H,m),6.94(0.5H,d,J=5.6Hz ),7.13(4H,t,J=8.5Hz),7.18-7.25(1.5H,m),7.40-7.48(4H,m),7.74(1H,t,J=5.6Hz).

[0449] MS (ESI) m / z (M+H) + :433.2.

[0450] Examples 121-138: Preparation of Compounds 121-138

[0451] Compounds 121 to 138 were prepared by using different intermediates using the method for preparing compound 1-B or 3. The intermediate numbers, structural formulas, MS and 1 The H-NMR data are shown in Table 14.

[0452] Table 14: Structures, MS and 1 H-NMR data

[0453]

[0454]

[0455]

[0456]

[0457]

[0458]

[0459]

[0460] Efficacy test

[0461] Test Example 1: In vitro BTK inhibition kinase activity test

[0462] 1: Compound preparation

[0463] Dissolve the compound powder in 100% DMSO to prepare a 10 mM stock solution and store it at -20 degrees in the dark.

[0464] 2: Kinase reaction process

[0465] (1) Prepare 1× Kinase buffer;

[0466] (2) Preparation of compound concentration gradient: The test compound was tested at a concentration of 1 μM and diluted to a 100% DMSO solution at 100 times the final concentration in the 384source plate. The compound was diluted 3 times to obtain 10 concentrations. 250 nL of the compound at 100 times the final concentration was transferred to the destination plate OptiPlate-384F using a dispenser Echo 550;

[0467] (3) Prepare a kinase solution with a final concentration of 2.5 times using 1× Kinase buffer;

[0468] (4) Add 10 μL of kinase solution at 2.5 times the final concentration to the compound wells and positive control wells respectively; add 10 μL of 1× Kinase buffer to the negative control wells;

[0469] (5) Centrifuge at 1000 rpm for 30 seconds, shake the reaction plate to mix, and incubate at room temperature for 10 minutes;

[0470] (6) Use 1× Kinase buffer to prepare a mixed solution of ATP and Kinase substrate 2 at 5 / 3 times the final concentration;

[0471] (7) Add 15 μL of a mixed solution of ATP and substrate at 5 / 3 times the final concentration to start the reaction;

[0472] (8) Centrifuge the 384-well plate at 1000 rpm for 30 seconds, shake to mix, and incubate at room temperature for 10 minutes;

[0473] (9) Add 30 μL of stop detection solution to stop the kinase reaction, centrifuge at 1000 rpm for 30 seconds, and shake to mix;

[0474] (10) Read the conversion rate using Caliper EZ Reader.

[0475] 3: Data Analysis

[0476] Calculation formula:

[0477]

[0478] Where: Conversion%_sample is the conversion rate reading of the sample; Conversion%_min: the mean of the negative control wells, representing the conversion rate reading of the wells without enzyme activity; Conversion%_max: the mean of the positive control well ratio, representing the conversion rate reading of the wells without compound inhibition.

[0479] Fitting the dose-effect curve: With the log value of the concentration as the X-axis and the percentage inhibition rate as the Y-axis, the log (inhibitor) vs. response-Variable slope of the analysis software GraphPad Prism5 was used to fit the dose-effect curve, thereby obtaining the IC50 value of each compound on the enzyme activity.

[0480] The calculation formula is Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope)).

[0481] The inhibitory activity of the compounds of the present invention on BTK wild type and BTK mutant C481S kinase is shown in Table 15:

[0482] IC50: A≤5nM; 5nM <B≤20nM;20nM<C≤100nM;100nM<D≤1000nM;

[0483] E>1000nM.

[0484] Table 15: Inhibitory activity of the compounds of the present invention on BTK and BTK-C481S kinases

[0485]

[0486]

[0487]

[0488]

[0489] “\” indicates that this test was not performed.

[0490] Test Example 2: Liver microsome stability test

[0491] 1: Add 10 μL of the test or control working solution and 80 μL of the microsome working solution (liver microsome protein concentration is 0.5 mg / mL) to the T0, T5, T10, T20, T30, T60 and NCF60 sample wells, and only add the microsome working solution to the Blank60 well. Then place the samples Blank60, T5, T10, T20, T30 and T60 except T0 and NCF60 in a 37°C water bath and pre-incubate for about 10 minutes;

[0492] 2: First add 300 μL of stop solution (containing 200 ng / mL tolbutamide and 200 ng / mL labetalol in acetonitrile) to the T0 sample and then add 10 μL of NADPH regeneration system working solution;

[0493] 3: After the pre-incubation of Blank60, T5, T10, T20, T30 and T60 plates, add 10 μL of NADPH regeneration system working solution to each sample well to start the reaction, and add 10 μL of 100 mM potassium phosphate buffer to the NCF60 sample well;

[0494] 4: After incubation for an appropriate time (e.g., 5, 10, 20, 30, and 60 minutes), add 300 μL of stop solution to each test sample well and control sample well on Blank60, T5, T10, T20, T30, T60, and NCF60 plates to terminate the reaction.

[0495] 5: All sample plates were shaken and centrifuged at 4000 rpm for 20 minutes. 100 μL of the test or reference supernatant was diluted into 300 μL of pure water for LC-MS / MS analysis.

[0496] 6: Data analysis, calculation of T based on first-order elimination kinetics 1 / 2 and CL int(mic) (μL / min / mg) value, the first-order elimination kinetic equation is:

[0497]

[0498] The results of stability tests on human and rat liver microsomes are shown in Table 16:

[0499] Table 16: Results of liver microsomal stability test of compounds of the present invention

[0500]

[0501]

[0502] Test Example 3: Pharmacokinetic Test

[0503] Each test compound was administered orally (10 mg / kg, 3 rats per group) once to SD rats for pharmacokinetic study. The test compound was dissolved with 5% DMSO + 10% solutol + 85% saline, and then vortexed for 1-2 minutes and ultrasonicated for 5-10 minutes to prepare a colorless, transparent and clear dosing solution. Animals were fasted overnight before oral administration, and food was resumed 4 hours after administration. After oral administration, SD rats were blood sampled from the eye sockets for pharmacokinetic samples. The collection time points were: 0.25h, 0.5h, 1h, 2h, 2.5h, 3h, 4h, 6h, 8h, and 10h after administration. Three whole blood samples were collected at each time point, and the collection volume was about 0.2-0.3mL. After blood samples were collected, they were immediately placed on ice and centrifuged to separate plasma within 15 minutes (centrifugation conditions: 8000rpm, 1 minute, room temperature). The collected plasma was stored at -20℃ before analysis. Take 20 μL of plasma sample into a 1.6 mL 96-well deep-well plate, add 200 μL of working internal standard solution (blank without internal standard is supplemented with the same volume of solvent), vortex mix for 1 min, centrifuge at 5800 rpm for 10 min, take 100 μL of supernatant and add it to a 96-well injection plate, and analyze by LC-MS / MS.

[0504] The pharmacokinetic test results of some compounds of the present invention are shown in Table 17 below:

[0505] Table 17 Pharmacokinetic test results of some compounds of the present invention

[0506]

[0507] Test Example 4: In vitro cell proliferation inhibition activity test

[0508] 1: Cell culture

[0509] The cells were cultured in 1640 medium supplemented with 10% inactivated FBS and 1% double antibody at 37°C and 5% CO2.

[0510] 2: Cell plating

[0511] (1) The cells are routinely cultured until the cell saturation reaches 80%-90%. When the cell quantity reaches the required level, the cells are harvested.

[0512] (2) Resuspend the cells in the appropriate culture medium, count the cells, and prepare a cell suspension of appropriate density.

[0513] (3) Add the cell suspension to a 96-well plate, 100 μL per well.

[0514] (4) The cells were cultured overnight in a 37°C, 5% CO2 incubator.

[0515] 3: Preparation of Compounds

[0516] (1) The test compounds were diluted with DMSO to a final concentration of 20 mM stock solution for later use.

[0517] (2) The stock solution was diluted 10-fold from 20 mM to 2 mM with DMSO, and then diluted 3-fold to 9 concentrations starting from 2 mM.

[0518] (3) The blank control wells were cells with 0.5% DMSO added, which served as high reading value control wells.

[0519] (4) Wells without cells but with culture medium were used as low reading control wells.

[0520] 4: Compound treatment of cells

[0521] (1) 24 hours after cell plating, add 99 μL of growth medium to each well, then add 1 μL of the compound prepared in step 4.3a), b), c), shake gently to ensure uniform mixing, and then place in a 37°C, 5% CO2 incubator.

[0522] (2) Place the cell plate in an incubator for 72 hours.

[0523] 5: CTG method detection

[0524] (1) The cell test plate was placed at room temperature for 30 minutes and 100 μL of culture medium was discarded from each well.

[0525] (2) Add 100 μL CTG reagent (CelltiterGlo kit) to each well, place on a fast shaker for 2 minutes, and place at room temperature away from light for 30 minutes.

[0526] (3) Read the chemiluminescent signal value using Envision instrument.

[0527] 6: Data Analysis

[0528] IC calculation using GraphPad Prism8 software 50 , the IC of the compound is obtained using the following nonlinear fitting formula 50 (half inhibitory concentration), the results are as follows:

[0529] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope))

[0530] X: log value of compound concentration, Y: inhibition rate (%inhibition)

[0531] Inhibition rate (%inhibition) = (high reading control reading - compound well reading) / (high reading control reading - low reading control reading) * 100

[0532] Table 18: Inhibitory activity of some compounds of the present invention on TMD8 cell proliferation

[0533]

[0534]

[0535] Table 19: Inhibitory activity of some compounds of the present invention on DOHH2 cell proliferation

[0536]

[0537] Table 20: Inhibitory activity of some compounds of the present invention on BT474 cell proliferation

[0538]

[0539] Table 21: Inhibitory activity of some compounds of the present invention on NCI-N87 cell proliferation

[0540]

[0541]

[0542] Test Example 5: HER2 kinase activity test

[0543] 1. Her2 kinase test steps

[0544] 1) Prepare 1× kinase reaction buffer: 1 volume of 5× kinase reaction buffer and 4 volumes of water, 1 mM dithiothreitol, 5 mM magnesium chloride, 1 mM manganese chloride, 12.5 mM SEB.

[0545] 2) Transfer 100 nl of the diluted compound working solution to each well of a reaction plate (784075, Greiner) using an Echo 550. Seal the reaction plate with a sealing film and centrifuge at 1000 g for 1 minute.

[0546] 3) Prepare 1 ng / μL Her2 kinase solution using 1× kinase reaction buffer.

[0547] 4) Add 5 μL of the above-prepared kinase solution to each well of the reaction plate. Seal the plate with a sealing film, centrifuge at 1000 g for 1 minute, and leave at room temperature for 10 minutes.

[0548] 5) Use 1× kinase reaction buffer to prepare a 2× kinase substrate and ATP mixture, where the 2× Her2 kinase substrate is 2 μM TK-substrate-biotin and 4 μM ATP.

[0549] 6) Add 5 μL of 2×TK-substrate-biotin and ATP mixture to the reaction plate and centrifuge at 1000 g for 30 seconds to start the reaction.

[0550] 7) Her2 kinase assay was carried out at room temperature for 50 minutes.

[0551] 8) Prepare a mixture of Sa-XL 665 (125 nM) and TK-antibody-Cryptate using HTRF detection buffer.

[0552] 9) Add 10 μL of Sa-XL 665 and TK-antibody-Cryptate mixture to each well, centrifuge at 1000 g for 30 seconds, and react at room temperature for 1 hour.

[0553] 10) Use Envision 2104 to read the fluorescence signals at 615 nm (Cryptate) and 665 nm (XL665).

[0554] 2. Data Analysis

[0555] 1) The inhibition percentage was calculated as follows:

[0556]

[0557] The average value of Ratio 665 / 615nm of all positive control wells in the whole plate.

[0558] The average value of Ratio 665 / 615nm of all negative control wells in the whole plate.

[0559] 2) Calculate IC50 and fit compound dose-effect curve:

[0560] The IC50 of the compound was obtained using the following nonlinear fitting formula using GraphPad 6.0.

[0561] Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope))

[0562] X: log value of compound concentration; Y: percentage of compound inhibition

[0563] Table 22: Inhibitory activity of some compounds of the present invention on HER2 kinase

[0564] Example HER2 IC50(nm) 130 4.35 145 1.69 146 3.92 152 4.19 155 9.44 Tucatinib 3.00

[0565] Test Example 6: Blood-brain barrier permeability test

[0566] Each test compound was subjected to a single oral pharmacokinetic study using SD rats at a dose of 10 mg / kg, with 9 animals in each group. The test compound was dissolved using 5% DMSO + 10% solutol + 85% saline, and a colorless, transparent, and clear dosing solution was prepared after vortexing for 1-2 minutes and ultrasonication for 5-10 minutes. The animals were fasted overnight before dosing. 3 SD rats were taken from each group at 1h, 2h, and 4h after dosing, and about 0.2-0.3mL of blood was collected from the eye sockets. After blood samples were collected, they were immediately placed on ice and centrifuged within 15 minutes to separate plasma (centrifugation conditions: 8000rpm, 1 minute, room temperature). The collected plasma was stored at -20°C before analysis. Cerebrospinal fluid and brain tissue were collected immediately after blood collection. Cerebrospinal fluid was extracted by puncturing the dura mater with a microinjector under direct vision. After chloral hydrate anesthesia, the head was fixed, the back hair was cut, a transverse incision (2 cm) was made at the line connecting the two ear roots, the muscle layer of the neck and the skull base was bluntly scraped to expose the foramen magnum, and about 100 μl of cerebrospinal fluid was taken with a 100 μl microinjector and stored at -20°C before analysis. The rats were immediately killed, decapitated, the brain tissue was dissected out, the surface capillaries were peeled off, weighed, and 3 times the amount of ice-cold physiological saline was added. The homogenizer was homogenized for 1 min and stored at -20°C before analysis. Take 20 μL of plasma sample and brain homogenate sample, add 200 μL of working internal standard solution (blank without internal standard, add the same volume of solvent), vortex mix for 1 min, centrifuge at 13500 rpm for 10 min, take 100 μL of supernatant, and analyze by LC-MS / MS. Take 20 μL of cerebrospinal fluid sample, add 60 μL of working internal standard solution (blank without internal standard, add the same volume of solvent), vortex mix for 1 min, centrifuge at 13500 rpm for 10 min, take 50 μL of supernatant, and analyze by LC-MS / MS.

[0567] Table 23: Blood-brain barrier permeability test results of some compounds of the present invention

[0568]

[0569] Test Example 7: TMD8 efficacy model test

[0570] Human diffuse large B-cell lymphoma TMD8 cells were cultured in vitro in a monolayer culture medium with RPMI1640 medium supplemented with 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin, and cultured in a 37°C 5% CO2 incubator. The cells were routinely digested and passaged twice a week using trypsin-EDTA. When the cell saturation reached 80%-90% and the number reached the required level, the cells were harvested, counted, and inoculated. 0.2 ml (1x10 7 TMD8 cells (with Matrigel, volume ratio of 1:1) were subcutaneously inoculated on the right back of each mouse, and the average tumor volume reached about 137 mm 3 The tumor diameter was measured with a vernier caliper twice a week. The formula for calculating the tumor volume was: V = 0.5a × b 2 , a and b represent the long and short diameters of the tumor, respectively.

[0571] Results Figure 1 and Figure 2 ;according to Figure 1 According to the TMD8 mouse subcutaneous transplant tumor efficacy model, under the same 10 mg / kg dosage condition, the tumor inhibition effects of the two compounds in Example 118 and Example 89-P1 were significantly better than those of the clinical phase II drug ARQ-531 and the marketed drug ibrutinib. Figure 2 , from the efficacy model of TMD8 mouse subcutaneous transplanted tumor, under the same dosage of 20 mg / kg, the inhibitory effects of Example 111-P1 and Example 125 on tumors were significantly better than those of Tirabrutinib. In particular, Example 111-P1, in terms of tumor inhibition rate, had a TGI of 93%, which was nearly 2 times that of Tirabrutinib, and almost completely controlled the growth of the tumor, with a very obvious efficacy advantage.

[0572] Test Example 8: DOHH-2-Luc Brain Tumor Efficacy Model Test

[0573] 1. Cell Culture

[0574] DOHH-2-luc tumor cells were cultured in vitro in RPMI 1640 medium containing 10% fetal bovine serum and 500ng / mL puromycin in an incubator at 37°C and 5% CO2. The medium was replenished or replaced every 2 to 3 days, and the number of passages did not exceed 4-5 times. Tumor cells in the logarithmic growth phase were used for inoculation of tumors in vivo.

[0575] 2. Tumor Cell Inoculation and Grouping

[0576] After intramuscular injection of Zota anesthesia, the animals were fixed on the operating table in a prone position. The skin on the top of the head was disinfected with iodine and 75% alcohol respectively. The skin was cut about 0.5 cm along the midline of the head to expose the coronal line and sagittal line. A brain locator was used to locate the position about 0.5-1.0 mm above the coronal line and about 2 mm to the right of the sagittal line, and a hole was drilled with a 1 mL syringe needle. The microinjector was vertically inserted into the location to a depth of 3 mm, and 3×10 DOHH-2-luc tumor cells were slowly injected (about 1 minute) 5 / 2μL suspension and leave the needle in place for 1 minute. After the needle is removed, the needle hole is quickly sealed with bone wax and the wound is sutured with a skin stapler. About 7 days after tumor inoculation, the animals were randomly divided into 5 groups, 5 in each group, based on animal weight and optical signal intensity at the tumor site.

[0577] 3. Imaging Analysis

[0578] The small animal in vivo imaging system IVIS Lumina III (Perkin Elmer) was used to image the mice 1-2 times a week according to their status, and the signal intensity of bioluminescence imaging (BLI, unit: photons / s) at the tumor cell inoculation site of the mice was monitored as the main indicator for evaluating tumor growth and drug efficacy. The specific operation was as follows:

[0579] After intraperitoneal injection of D-luciferin (15 mg / mL, 5 μL / g according to the weight of the experimental animal), the mice were anesthetized by inhalation of 1%-2% isoflurane. Ten minutes after the injection of D-luciferin, the animals were imaged using IVIS Lumina III. The data were analyzed and processed using Living Image software (Perkin Elmer) to calculate the optical signal intensity within the ROI (regions of interest) of each animal.

[0580] Results Figure 3 , Figure 4 ;according to Figure 3 In the study of DOHH2 tumor model in mouse brain, under the same dosage of 30 mg / kg (BID), the tumor inhibition effects of Example 111-P1 and Example 125 were significantly better than those of Tirabrutinib, and the efficacy advantage was very obvious, and no side effects were found after 21 days of administration.

[0581] Figure 4The fluorescence images of all the test animals after imaging are shown in Figure 1. The image uses color and area size to indicate the size of the tumor in the brain. The redder the color, the larger the tumor. As can be seen from the image, at the same dose, Example 111-P1 and Example 125 have very good inhibitory effects on tumors, with almost no red areas, indicating that the brain tumors of the animals in these two groups are very small; while all animals in the model group and Tirabrutinib group have large red areas, indicating that the tumors are very large.

[0582] As can be seen from the above examples, the compounds of the present invention as BTK protein kinase inhibitors have a structure of formula I, preferably a structure of formula II; they have strong inhibitory effects on wild-type BTK and mutant BTK (C481S), and have good pharmacokinetic properties, and can be used to prepare drugs for treating diseases caused by overexpression of BTK kinase. Some of the compounds are significantly better than the marketed BTK inhibitors Ibrutinib, Tirabrutinib and ARQ-531 in clinical phase II in the TMD8 subcutaneous tumor efficacy model experiment.

[0583] Some compounds of the present invention are significantly superior to the marketed drugs Tirabrutinib and Tucatinib in terms of blood-brain barrier permeability, liver microsome stability, and pharmacokinetic properties. In the DOHH-2-Luc brain efficacy model, some compounds have very good efficacy, which also verifies the brain permeability data. They can be used to prepare drugs for treating diseases caused by overexpression of BTK or HER2 kinases, especially brain diseases.

[0584] The compounds described above in the present invention or their stereoisomers, solvates, hydrates, pharmaceutically acceptable salts or cocrystals can be used to prepare drugs for treating any one or more of autoimmune diseases, inflammatory diseases, thromboembolic diseases, allergies, infectious diseases, proliferative disorders and cancers, and are expected to provide new and good treatment options.

[0585] The above descriptions are only preferred embodiments of the present invention. It should be pointed out that professionals in this technical field can implement various modifications to these embodiments without departing from the technical principles of the present invention, and these modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A compound as a BTK inhibitor, characterized in that Having the structure shown in Formula III, Formula IV, or a pharmaceutically acceptable salt thereof: Wherein, R1 is selected from hydrogen, amino or methyl; R2 is selected from hydrogen, halogen or methyl; R3, R4 are selected from hydrogen, methyl, ethyl or isopropyl; R6 is selected from hydrogen, fluorine, chlorine, bromine, trifluoromethyl, methyl, methoxy, trifluoromethoxy, difluoromethoxy; m is selected from 0, 1, 2; n is selected from 0 and 1; n1 is selected from 0, 1, 2; X is selected from n2 is selected from 0, 1, 2, 3, 4; R8 is independently selected from fluorine, chlorine, bromine, cyano, amino, C1-C3 alkyl, C1-C3 alkoxy, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, trifluoromethyl, trifluoromethoxy, difluoromethoxy, deuterated methoxy; But not including compounds (5-(8-amino-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; (5-(8-amino-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; (5-(8-amino-1-(2,3-difluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; (5-(8-amino-1-(4-(2,3-difluorophenoxy)phenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; and the trans isomers of:

2. The compound according to claim 1, characterized in that R8 is independently selected from fluoro, chloro, bromo, cyano, methoxy, cyclopropyl, ethyl, isopropyl.

3. A compound as a BTK inhibitor, characterized in that The compound structure is selected from one of the following:

4. A pharmaceutical composition, characterized in that The active ingredient of the pharmaceutical composition is selected from one or a combination of two or more of the compounds or pharmaceutically acceptable salts thereof according to claim 1.

5. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a protein kinase inhibitor, wherein the protein kinase inhibitor is a BTK inhibitor.

6. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating diseases caused by overexpression of BTK kinase.

7. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating any one or more of autoimmune diseases, inflammatory diseases, thromboembolic diseases, allergies, infectious diseases, proliferative disorders and cancers caused by overexpression of BTK kinase.

8. The use according to claim 7, characterized in that The disease is selected from the group consisting of arthritis, rheumatoid arthritis, urticaria, vitiligo, organ transplant rejection, ulcerative colitis, Crohn's disease, dermatitis, asthma, Sjögren's syndrome, systemic lupus erythematosus, multiple sclerosis, idiopathic thrombocytopenic purpura, rash, anti-neutrophil cytoplasmic antibody vasculitis, pemphigus, chronic obstructive pulmonary disease, psoriasis; mantle cell lymphoma, ovarian cancer, esophageal cancer, laryngeal cancer, glioblastoma, neuroblastoma, cell carcinoma, gastric cancer, hepatocellular carcinoma, glioma, melanoma, kidney cancer, bladder cancer, bile duct cancer, lymphoma, hairy cell carcinoma, nasopharyngeal cancer, pharyngeal cancer, colorectal cancer, rectal cancer, brain and central nervous system cancer, genitourinary tract cancer, lung cancer, small cell carcinoma, bone cancer, colon cancer, adenoma, follicular carcinoma, Hodgkin's leukemia, bronchial cancer, multiple myeloma, lymphocytic leukemia, myeloid leukemia, non-Hodgkin's lymphoma, primary macroglobulinemia.

9. The use according to claim 7, characterized in that: The disease is selected from: non-small cell lung cancer.

10. The use according to claim 7, characterized in that The disease is selected from the group consisting of: pemphigus vulgaris, prostate cancer, testicular cancer, endometrial cancer, uterine body cancer, cervical cancer, adenocarcinoma, acute myeloid leukemia, chronic myeloid leukemia, and chronic lymphoid leukemia.

11. The use according to claim 7, characterized in that: The disease is selected from the group consisting of breast cancer, lung adenocarcinoma, thyroid cancer, and pancreatic cancer.

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