Compounds as BTK inhibitors, their production methods and applications

Abstract

The present invention provides a compound as a BTK inhibitor, and a method for preparing and using the same, wherein the compound has the structure of Formula I, or in the form of its tautomer, meso isomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable hydrate, solvate, or salt. In the formula, A1, A2, A3, A4, A5, and A6 are each independently selected from C-R5 or N, at least one of which is N; and M is selected from a substituted or unsubstituted saturated hydrocarbon group or heterosaturated hydrocarbon group, a substituted or unsubstituted unsaturated cyclic group or heterocyclic group, or a substituted or unsubstituted monocyclic, bicyclic, or tricyclic aryl group or heteroaryl group. The BTK protein kinase inhibitor of the present invention has potent inhibitory activity against wild-type BTK and mutant BTK (C481S), favorable pharmacokinetic properties, and promising application prospects. TIFF2023533349000169.tif66170

Description

cross reference

[0001] This application claims priority to a Chinese patent application filed on July 15, 2020 with the China Patent Office, application number 202010679776.6, entitled "Compound as BTK inhibitor, and preparation method and use thereof", and also claims priority to a Chinese patent application filed on November 25, 2020 with the China Patent Office, application number 202011337022.9, entitled "Compound as BTK inhibitor, and preparation method and use thereof", the entire contents of which are incorporated herein by reference. [Technical field]

[0002] The present invention relates to the technical field of medicine, in particular to compounds as BTK protein kinase inhibitors and their preparation methods and uses. [Background technology]

[0003] Bruton's tyrosine protein kinase (BTK) is a member of the non-receptor protein tyrosine kinase Tec family, which is mainly expressed in various hematopoietic cell lines. The Tec family is the second largest family of human non-receptor kinases after the Src family, and includes BTK, BMX (etk), ITK, TEC, and TXK (RLK) as its main members. In 1993, BTK was identified as a defective protein in human X-linked agammaglobulinemia (X-1-inked agammaglobulinemia, XLA). BTK is a key regulator of the B cell receptor (BCR) signaling pathway, plays a key role in the activation, proliferation, differentiation, and survival of B cells, and is closely associated with various B cell tumors and autoimmune diseases.

[0004] The BTK structure contains five major domains: the PH domain (pleckstrin homology), the TH domain (Tec homology), the SH3 domain (Src homology 3), the SH2 domain (Src homology 2), and the SHl domain (Src homology 1). Activation (phosphorylation) of BTK occurs first at the activation ring of the SHl domain, and then at the SH2 and SH3 domains, which contain the major autophosphorylation sites. These SH domains also contain the nuclear localization signal (NLS) and nuclear export signal (NES) required for BTK to perform nucleocytoplasmic shuttling.

[0005] BTK plays an irreplaceable role in B lymphopoiesis and can control B cell development and differentiation by activating cell cycle positive regulators and differentiation factors, and can also control B cell survival and proliferation by regulating the expression of pro- and anti-apoptotic proteins. Persistent activation of BTK is a prerequisite for the development of chronic lymphocytic leukemia (CLL), and aberrant BCR-BTK signaling may promote the survival of activated B cell subtypes in diffuse large B cell lymphoma (DLBCL). Gain-of-function mutations of BTK have also been identified in colorectal cancer, acute lymphocytic leukemia (ALL), and chronic granulocytic leukemia (CML). Thus, it has been demonstrated that aberrant activation of BTK-dependent pathways is closely related to the development and progression of various tumors.

[0006] Currently, approved irreversible BTK inhibitors include, for example, ibrutinib, acalabrutinib, and zanubrutinib, all of which selectively form irreversible covalent bonds with the cysteine ​​residue (Cys-481) of BTK to inhibit BTK activity and treat related diseases. However, some cancer patients may develop resistance to first-generation BTK inhibitors, creating new unmet clinical needs. Research has shown that BTK-C481S mutation is one of the main mechanisms of drug resistance, so drugs that can target and inhibit BTK-C481S mutation are expected to provide a new treatment regimen. For example, ARQ-531 is an orally bioavailable, effective, and reversible dual BTK inhibitor against wild-type and C481S mutations, and initial clinical results of ARQ-531 have shown that it is effective in patients with C481S mutations. Summary of the Invention [Problem to be solved by the invention]

[0007] The present invention has been made in view of the above circumstances, and provides a compound as a BTK inhibitor, and a method for producing and using the same. The compound provided by the present invention can be used as a BTK protein kinase inhibitor, and has characteristics such as high inhibitory activity.

[0008] The present invention provides a compound having the structure shown in Formula I below, or in the form of a tautomer, meso isomer, racemate, enantiomeric isomer, diastereomeric isomer, or mixture thereof, a pharma- ceutically acceptable hydrate, solvate, or salt thereof.

[0009] [ka]

[0010] wherein A1, A2, A3, A4, A5, and A6 are each independently selected from C-R5 or nitrogen (N), and at least one of A1, A2, A3, A4, A5, and A6 is N; M is selected from a substituted or unsubstituted saturated hydrocarbon group or heterosaturated hydrocarbon group, a substituted or unsubstituted unsaturated cyclic group or heterocyclic group, and a substituted or unsubstituted monocyclic, bicyclic or tricyclic aryl group or heteroaryl group, each of which is independently selected from an aryl group or heteroaryl group substituted with an arbitrary group, an alkyl group or heteroalkyl group, a cycloalkyl group or heterocycloalkyl group, an unsaturated cyclic group or heterocyclic group, a phenoxy group, a halogen, a hydroxy group, a cyano group, an amino group, an ester group, a nitro group, a mercapto group, an amide group, a sulfonyl group, a phosphonyl group, an alkyloxyphosphorus group, an alkylsulfone group, and an alkylsulfoxide group, and the substitution substituent is an aryl group or heteroaryl group substituted with an arbitrary group, or a phenyl group substituted with an arbitrary group; Q is CR 10 R 11 , N.R. 12 , oxygen (O), sulfur (S), S(O), and S(O)2; R1, R2, R3, R4, R5, R 10 , R 11 , R 12are each independently selected from hydrogen, deuterium, halogen, a substituted or unsubstituted alkyl group or heteroalkyl group, a substituted or unsubstituted cycloalkyl group or heterocycloalkyl group, a substituted or unsubstituted unsaturated cyclic group or heterocyclic group, a substituted or unsubstituted aryl group or heteroaryl group, a hydroxy group, a cyano group, an amino group, an ester group, a nitro group, a mercapto group, an amido group, a sulfonyl group, a phosphonyl group, an alkyloxyphosphorus group, an alkylsulfone group, and an alkylsulfoxide group; or R3 and R4 together with the carbon atoms adjacent thereto form a substituted or unsubstituted C3-C10 cycloalkyl group or heterocycloalkyl group, and the substituent of the substitution is selected from a halogen, a hydroxy group, a cyano group, an amino group, a mercapto group, a nitro group, a carboxy group, a hydroxyamino group, an alkyl group, a cycloalkyl group, a heteroalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, an ester group, an acyl group, an amido group, a sulfonyl group, and a phosphonyl group; m is selected from integers of 0 to 6; n is selected from integers of 0 to 3.

[0011] The compounds of the present invention may be in any form having the structure of Formula I, including tautomers, meso isomers, racemates, enantiomers, diastereoisomers, or mixtures thereof, pharma- ceutically acceptable hydrates, solvates, or salts.

[0012] In the present invention, "selected from the group" generally means that the members of the group are in a parallel relationship. In the structure represented by formula I, preferably, three or four of A1, A2, A3, A4, A5, and A6 are N, and the position of R2 is not limited, and the para position of R1 is preferable. In the present invention, the substitution may be mono- or poly-substituted (e.g., di- or tri-substituted), 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 may have one or more carbon atoms substituted with a heteroatom, and the heteroatom is an atom other than carbon (C), such as oxygen, sulfur, nitrogen, or phosphorus (P). In addition, the halogen includes fluorine (F), chlorine (Cl), bromine (Br), and the like, and fluorine or chlorine is preferable. The above "C3 to C10" is an integer having a carbon number selected from 3 to 10. Hereinafter, the same expression will not be repeated.

[0013] In the present invention, the ring system formed by the bridging atom being bonded to the ring by chemical bond (shown in the following formula) means that the bridging atom can be connected to any available C atom on the ring to form any spiro ring or bridged ring structure compound. For example, the following formula shows that the bridging atom Q can be connected to any available C atom on the 6-membered ring, that is, the bridging atoms are connected to the same C atom to form a spiro compound, for example, the bridging atoms are connected to the second C atom or all are connected to the third C atom, and the bridging atoms are connected to different C atoms to form a bridged ring compound, for example, the bridging atoms are connected to the first and fourth C atoms or the second and fourth C atoms, respectively.

[0014] [ka]

[0015] Preferably, the compound has the structure shown in Formula II, or in the form of a tautomer, meso isomer, racemate, enantiomeric isomer, diastereomeric isomer, or mixture thereof, a pharma- ceutically acceptable hydrate, solvate, or salt thereof.

[0016] [ka]

[0017] In the formula, R1 is selected from hydrogen, halogen, a hydroxy group, a cyano group, an amino group, a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group, a substituted or unsubstituted C1-C6 heteroalkyl group, and a substituted or unsubstituted C3-C6 heterocycloalkyl group; R1 is selected from hydrogen, an amino group, a methyl group, an ethyl group, a methoxy group, a cyano group, a trifluoromethyl group, an isopropyl group, and a cyclopropyl group; R1 is further selected from hydrogen (H), an amino group (NH2), and a methyl group (CH3); R2 is selected from hydrogen, halogen, a hydroxy group, a cyano group, an amino group, a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group, a substituted or unsubstituted C1-C6 heteroalkyl group, and a substituted or unsubstituted C3-C6 heterocycloalkyl group; R2 is selected from hydrogen, fluorine, chlorine, bromine, a methyl group, an ethyl group, a methoxy group, a cyano group, a trifluoromethyl group, an isopropyl group, and a cyclopropyl group; R2 is further selected from hydrogen, chlorine, and a methyl group; R3 and R4 are selected from hydrogen, a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group, a substituted or unsubstituted C1-C6 heteroalkyl group, and a substituted or unsubstituted C3-C6 heterocycloalkyl group, or R3 and R4 together with the carbon atoms connected thereto form a substituted or unsubstituted C3-C6 cycloalkyl group or a heterocycloalkyl group containing N and O atoms; Further, R3 and R4 are selected from hydrogen, a methyl group, an ethyl group, an isopropyl group, and a cyclopropyl group, or R3 and R4 together with the carbon atom connected thereto form a cyclopropyl group, an azetidinyl group, an azacyclopentyl group, an azacyclohexyl group, an oxetanyl group, an oxacyclopentyl group, or an oxacyclohexyl group; R6 is selected from hydrogen, halogen, a hydroxyl group, a cyano group, an amino group, a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group, a substituted or unsubstituted C1-C6 heteroalkyl group, and a substituted or unsubstituted C3-C6 heterocycloalkyl group; R6 is selected from hydrogen, halogen, a cyano group, a substituted or unsubstituted C1-C3 alkyl group, and a substituted or unsubstituted C1-C3 alkyloxy group; R6 is selected from hydrogen, fluorine, chlorine, bromine, a trifluoromethyl group, a methyl group, a methoxy group, a trifluoromethoxy group, and a difluoromethoxy group; and R6 is hydrogen or fluorine.

[0018] m is selected from 0, 1, 2, 3; n is selected from 0, 1, 2; n1 is selected from 0, 1, 2, 3, 4; R7 is selected from a substituted or unsubstituted aryl group and a substituted or unsubstituted pyridyl group, the substituents of the substitution are independently selected from halogen, hydroxyl group, amino group, cyano group, alkyl group, heteroalkyl group, cycloalkyl group, and heterocycloalkyl group, the substituents are independently selected from fluorine, chlorine, bromine, cyano group, amino group, C1-C3 alkyl group, C1-C3 alkyloxy group, C3-C6 cycloalkyl group, and C3-C6 heterocycloalkyl group, the substituents are independently selected from fluorine, chlorine, bromine, cyano group, trifluoromethyl group, trifluoromethoxy group, difluoromethoxy group, methoxy group, deuterated methoxy group, cyclopropyl group, cyclopropylmethoxy group, ethyl group, isopropyl group, and isobutyl group, where the number of the substituents is an integer between 0 and 5,

[0019] [ka]

[0020] In some embodiments, X is selected from acceptable linking groups such as

[0021] [ka]

[0022] However, R9, R 13 are independently selected from hydrogen, halogen, a hydroxy group, an amino group, a cyano group, a C1-C3 alkyl group, a C1-C3 alkyloxy group, a C3-C6 cycloalkyl group, and a C3-C6 heterocycloalkyl group; or R and R 13 form, together with the carbon atoms connected thereto, a substituted or unsubstituted C3-C6 cycloalkyl group or a substituted or unsubstituted C3-C6 heterocycloalkyl group containing N or O; and R9 and R 13 are independently selected from hydrogen, fluorine, chlorine, cyano, methyl, ethyl, isopropyl, cyclopropyl, trifluoromethyl, and isobutyl; or R and R 13 together with the carbon atoms associated therewith form a cyclopropyl group, and R and R 13 is selected from hydrogen, fluorine, deuterium, chlorine, a methyl group, a hydroxyl group, and an amino group.

[0023] [ka]

[0024] [ka]

[0025] All compounds represented by the formula: R can be used as brain-penetrant BTK inhibitors or HER2 inhibitors. 13 Both are fluorine.

[0026] In some embodiments of the invention, the compound has the structure shown in Formula III, Formula IV, or in the form of a tautomer, meso-isomer, racemate, enantiomeric isomer, diastereomeric isomer, or mixture thereof, a pharma- ceutically acceptable hydrate, solvate, or salt thereof.

[0027] [ka]

[0028] In the formula, the structures of R1, R2, R3, R4, R6, and X are as described above, and m, n, and n1 are also as described above. For example, X is

[0029] [ka]

[0030] In formulas III to IV, n2 is selected from 0, 1, 2, 3, and 4; R8 is independently selected from hydrogen, halogen, hydroxyl, amino, cyano, alkyl, heteroalkyl, cycloalkyl, and heterocycloalkyl groups; R8 is independently selected from hydrogen, fluorine, chlorine, bromine, cyano, amino, C1-C3 alkyl, C1-C3 alkyloxy, C3-C6 cycloalkyl, and C3-C6 heterocycloalkyl; and the substituents are independently selected from hydrogen, fluorine, chlorine, bromine, cyano, trifluoromethyl, trifluoromethoxy, difluoromethoxy, methoxy, deuterated methoxy, cyclopropyl, cyclopropylmethoxy, ethyl, isopropyl, and isobutyl; provided that the number of the substituents is an integer between 0 and 5 (including the end points), and the multiple substituents may be the same or different; and in formula IV, the substituted or unsubstituted pyridyl group is not limited in its bonding position and may be bonded to the ortho position of N.

[0031] In some embodiments of the present invention, the N-containing fused ring of Formula II-IV is

[0032] [ka]

[0033] and the single bonds at both ends of the ring are bonds for linking. In the X structures in formulae II to IV, the curved single bond represents a bond for linking. The position of R6 is not limited. n1 is preferably 0, 1 or 2. When n=0, it is a 5-membered ring, and when n is 1, it is a 6-membered ring, which can be inferred.

[0034] Preferably, in formulas II to IV, R1 is an amino group, R2 is hydrogen or chlorine, R6 is hydrogen or monosubstituted fluorine, R7 in formula II is a substituted or unsubstituted phenyl group or pyridyl group, 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 group, or form a cyclopropyl group together with the carbon atom connected thereto.

[0035] Specifically, the structure of the compound described in the present application is selected from any one of the following (however, the one having a single bond at one end is a methyl group as shown in formula 5 of compound 5), and is also any of the compounds represented by 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, and has excellent performance.

[0036] [ka]

[0037] [ka]

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[0039] [ka]

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[0050] [ka]

[0051] [ka]

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[0053] [ka]

[0054] The present invention provides a pharmaceutical composition, the active ingredient of which is one or a combination of two or more selected from the above-mentioned compounds or their stereoisomers, solvates, hydrates, pharma- ceutically acceptable salts, or cocrystals. Furthermore, the present invention is not particularly limited with respect to the type of formulation of the pharmaceutical composition.

[0055] The present invention provides the use of the aforementioned compound or a stereoisomer, solvate, hydrate, pharma- ceutically acceptable salt or cocrystal thereof in the manufacture of a protein kinase inhibitor, wherein the kinase inhibitor is a BTK inhibitor or a HER2 inhibitor. Alternatively, the present invention provides the use of the aforementioned compound or a stereoisomer, solvate, hydrate, pharma-ceutically acceptable salt or cocrystal thereof in the manufacture of a medicament for treating a disease caused by overexpression of BTK kinase or HER2 kinase.

[0056] The present invention provides the use of the aforementioned compound or a stereoisomer, solvate, hydrate, pharma- ceutically acceptable salt or co-crystal thereof in the manufacture of a medicament for treating any one or more diseases selected from an autoimmune disease, an inflammatory disease, a thromboembolic disease, an allergy, an infectious disease, a proliferative disease and cancer.

[0057] Further, the diseases include arthritis, rheumatoid arthritis, urticaria, vitiligo, organ transplant rejection, ulcerative colitis, Crohn's disease, dermatitis, asthma, Sjogren's syndrome, systemic lupus erythematosus, multiple sclerosis, idiopathic platelet-reduced purpura, skin rash, antineutrophil cytoplasmic antibody-associated vasculitis, pemphigus, 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. , renal cancer, pancreatic cancer, lymphoma, hairy cell carcinoma, nasopharyngeal cancer, pharyngeal cancer, colon 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 carcinoma, lung adenocarcinoma, bone cancer, colon cancer, adenoma, pancreatic cancer, adenoma, thyroid cancer, follicular carcinoma, Hodgkin's leukemia, bronchial cancer, thyroid cancer, uterine cancer, cervical cancer, multiple myeloma, acute myeloid leukemia, chronic myeloid leukemia, lymphocytic leukemia, chronic lymphocytic leukemia, osteomyeloma, non-Hodgkin's lymphoma, primary macroglobulinemia.

[0058] In the prior art, the inhibitory activity of ARQ-531 has room for improvement, and its inhibitory activity against cells such as TMD8 and REC-1 is low, leading to problems such as excessive clinical doses and high side effects. In addition, ARQ-531 has poor selectivity, high inhibitory activity against TEC and EGFR, which is prone to side effects such as bleeding, diarrhea, and eczema. Furthermore, its pharmacokinetics is not very ideal, and in preclinical studies, the bioavailability was only 38% in dog PK experiments. Thus, ARQ-531 has a lot of room for improvement in terms of inhibitory activity, selectivity, and pharmacokinetics.

[0059] In the in vitro BTK inhibition and HER2 inhibition kinase activity assay in the embodiment of the present invention, the powder of the compound can be dissolved in 100% DMSO and prepared into a 10 mM stock solution, and frozen and stored at -20 degrees in the dark. During the kinase reaction, the test concentration of the test compound is 1 μM, and it is diluted into a 100% DMSO solution at 100 times the final concentration in a 384 source plate, and the compound is diluted 3 times to a concentration of 10. In addition, in the embodiment of the present invention, the compound was also used to perform experiments such as liver microsome metabolic stability, rat PK, rat brain penetration rate, and drug effect model. Compared with the existing clinical drug (ARQ-531), the compound of the present invention has advantages as a BTK protein kinase inhibitor in BTK, BTK (C481S) inhibitory activity, liver microsome metabolic stability, pharmacokinetics in rats, and toxicity. Compared with the existing commercially available drug tirabrutinib, the compound of the present invention, as a BTK protein kinase inhibitor, has advantages in BTK, BTK(C481S) inhibitory activity, cellular activity, liver microsomal metabolic stability, pharmacokinetics in rats, and rat blood-brain barrier permeability.

[0060] In an embodiment of the present invention, the target compound is designed and synthesized by the manufacturing process shown below. Specifically, intermediate A (also referred to as a boronic acid or boronic ester compound represented by formula A) and intermediate B (a bromo compound represented by formula B) are subjected to Suzuki reaction to synthesize intermediate C (an intermediate represented by formula C), which is then deprotected to obtain a compound having a structure represented by formula II. In a specific embodiment, intermediate C is obtained by coupling a commercially available boronic acid A or a self-made boronic ester A with a self-made bromo compound B under palladium catalysis, and intermediate C is deprotected to obtain the compound of the embodiment. Compared with the ARQ-531 drug in phase II clinical trials, the compound of the present invention has obvious improvements in the inhibitory activity of BTK and BTK (C481S), liver microsomal metabolic stability, and pharmacokinetics in rats.

[0061] Furthermore, the following synthetic methods according to the embodiments of the present invention are simple and have high yields.

[0062] [ka]

[0063] An embodiment of the present invention relates to a method for producing the above-mentioned BTK inhibitor, comprising:

[0064] [ka]

[0065] (where R1, R2, R3, R4, m, and n are as described above), for example,

[0066] [ka]

[0067] The present invention provides an intermediate compound having the structure:

[0068] Other intermediate compounds include:

[0069] [ka]

[0070] etc. [Brief description of the drawings]

[0071] [Figure 1] FIG. 1 shows the test results of some compounds of the present invention using the TMD8 drug effect model. [Diagram 2] FIG. 2 shows the test results of some compounds of the present invention using the TMD8 drug effect model. [Diagram 3] FIG. 3 shows the test results of some compounds of the present invention using a DOHH-2-Luc brain tumor drug effect model. [Figure 4] FIG. 4 shows fluorescence photographs of the test results of some compounds of the present invention using a DOHH-2-Luc brain tumor drug effect model. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0072] The following clearly and completely describes the technical solutions in the embodiments of the present invention, but it is clear that the described embodiments are only some of the embodiments of the present invention, and are not all of the embodiments. All other embodiments that can be obtained by those skilled in the art based on the embodiments of the present invention without creative efforts fall within the scope of the present invention.

[0073] In order to further understand the present invention, the compounds provided in the present application that can be used as BTK protein kinase inhibitors, as well as methods for producing and using the same, are specifically described below with reference to examples.

[0074] In an embodiment of the present invention, the structure of a compound can be determined by mass spectrometry (MS) or nuclear magnetic resonance (NMR) 1 The temperature was determined by a 1 H NMR (H NMR) instrument. The term "room temperature" refers to a temperature between 10° C. and 25° C. The chemical abbreviations have the following meanings: DMF: N,N-Dimethylformamide DIEA: N,N-Diisopropylethylamine HATU: O-(7-azabenzotriazol-1-yl)-N,N,N'-tetramethyluronium hexafluorophosphate PdCl2(dppf):1,1-bis(diphenylphosphino)ferrocene]dichloropalladium DCM: dichloromethane TEA: triethylamine TBDPSCl: Lithium bistrimethylsilylamide 9-BBN: 9-Borabicyclo[3.3.1]nonane Dess-Martin: Dess-Martin oxidizer DME: Dimethoxyethane TosMIC: p-Toluenesulfonylmethylisocyanide t-BuOK: Potassium tert-butoxide Dibal-H: Diisobutylaluminum hydride THF: Tetrahydrofuran NBS: N-bromosuccinimide TBAF: Tetrabutylammonium fluoride DMSO: Dimethyl sulfoxide LDA: Lithium diisopropylamide HBTU: Benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate NMP: N-methyl-2-pyrrolidone BAST: Bis(2-methoxyethyl)aminosulfur trifluoride PMDTA: Pentamethyldiethylenetriamine DMA: N,N-Dimethylacetamide dppf: 1,1'-bis(diphenylphosphino)ferrocene Pd2(dba)3: Tris(dibenzylideneacetone)dipalladium TsCl: 4-toluenesulfonyl chloride DMAP: 4-dimethylaminopyridine PDC: Pyridinium dichromate DIAD: diisopropyl azodicarboxylate; NCS: N-chlorosuccinimide.

[0075] Preparation of intermediate A-1:

[0076] [ka]

[0077] 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 flask, the reaction liquid was replaced with nitrogen, cooled to 0 ° C, HATU (24.2 g, 63.7 mmol) was added in several portions, the reaction mixture was slowly warmed to room temperature, and the reaction was stirred overnight, and TLC showed that the raw material reaction was complete. Water was added to the reaction system, and it was extracted twice with ethyl acetate, and the organic phase was combined, washed with water, washed with saturated saline, dried with anhydrous Na2SO4, evaporated in vacuum, and then purified with a silica gel column to obtain 11.9 g of product A-1-3, with a yield of 76%.

[0078] Compound A-1-3 (5.0 g, 16.9 mmol), dioxane (50 mL), bis(pinacolato)diboron (5.2 g, 20.3 mmol) and potassium acetate (2.5 g, 25.4 mmol) were added to the flask, 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, and the raw material reaction was complete by TLC. After cooling the reaction system, silica gel was directly added and stirred, and then purified with a silica gel column to obtain a crude product, which was slurried with petroleum ether to obtain 3.4 g of product A-1, with a yield of 62%.

[0079] Preparation of intermediate A-2:

[0080] [ka]

[0081] Compound A-2-1 (2.0 g, 11.8 mmol) and dioxane (20 mL) were added to a flask and cooled in an ice-water bath. Hydrogen peroxide (20 mL, 30%) was added dropwise to the reaction solution, and the reaction solution was stirred at room temperature overnight to stop the reaction. Water was added to the reaction system, and the mixture was extracted four times with ethyl acetate. The organic phases were combined, washed with water, washed with saturated saline, evaporated in vacuum, and then purified with a silica gel column to obtain 1.6 g of product A-2-2, with a yield of 96%.

[0082] 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 a flask, and the reaction solution was replaced with nitrogen, then heated to 140 ° C. and stirred overnight to react. The reaction system was cooled to room temperature, water was added, and the mixture was extracted three times with ethyl acetate. The organic phase was combined, washed with water, washed with saturated saline, evaporated in vacuum, and purified with a silica gel column to obtain 800 mg of product A-2-3, with a yield of 38%.

[0083] Compound A-2-3 (800 mg, 2.69 mmol), dioxane (16 mL), bis(pinacolato)diboron (821 mg, 3.23 mmol) and potassium acetate (528 mg, 5.38 mmol) were added to a flask, 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 cooling the reaction system, silica gel was directly added and mixed, and then purified with a silica gel column to obtain 520 mg of product A-2, with a yield of 56%.

[0084] Preparation of intermediate A-3:

[0085] [ka]

[0086] 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 sieves (1.66 g) were added to a flask, and the reaction solution was stirred at room temperature overnight to react. The reaction solution was filtered by suction, and silica gel was added to the filtrate and mixed, and then purified with a silica gel column to obtain 600 mg of product A-3-3, with a yield of 43%.

[0087] The synthesis method of A-3 was carried out in accordance with the synthesis method of A-2 from A-2-3. Preparation of intermediate A-4:

[0088] [ka]

[0089] 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 flask, and the reaction solution was heated to 100 ° C. and stirred overnight to react, and TLC showed that the raw material reaction was complete. Water was added to the reaction system, and it was extracted twice with ethyl acetate, and the organic phase was combined and washed with water, washed with saturated saline, dried with anhydrous Na2SO4, and evaporated in vacuum 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.

[0090] The synthesis method of A-4 was carried out in accordance with the synthesis method of A-2 from A-2-3.

[0091] The following compounds were synthesized using the corresponding commercially available starting materials by the methods for preparing the above intermediates.

[0092] [Table 1]

[0093] Preparation of intermediate A-13:

[0094] [ka]

[0095] 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 flask, the reaction solution was cooled to -70 ° C., and LDA (2 m, 3.3 mL, 6.6 mmol) was added dropwise to the reaction solution. After the addition was completed, 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 saline, spin-dried, and the residue was purified with a silica gel column to obtain 838 mg of product A-13-2, with a yield of 67%.

[0096] The synthesis method of A-13 was carried out in accordance with the synthesis method of A-2 from A-2-1. Preparation of intermediate A-14:

[0097] [ka]

[0098] 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 a flask, and the reaction solution was heated to 50° C. and stirred overnight. The reaction solution was poured into water, adjusted with dilute hydrochloric acid, extracted twice with ethyl acetate, and the combined organic phase was washed with saturated saline and spin-dried. The residue was purified on a silica gel column to obtain 432 mg of product A-14-2, with a yield of 76%.

[0099] The synthesis method of A-14 was carried out in accordance with the synthesis method of A-2 from A-2-2. Preparation of Intermediate A-15:

[0100] [ka]

[0101] Compound A-2-3 (500 mg, 1.68 mmol) and dichloromethane (8 mL) were added to the 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, and the reaction was allowed to proceed for 2 hours after completion of the addition. Water was added to the reaction solution to quench the reaction, and the reaction solution was extracted twice with dichloromethane. The organic phases were combined and spin-dried, and the residue was purified with a silica gel column to obtain 390 mg of product A-15-1, with a yield of 82%.

[0102] Compound A-15-1 (200 mg, 0.706 mmol), DMF (2 mL), bromocyclopropane (171 mg, 1.41 mmol), cesium carbonate (276 mg, 0.847 mmol) and sodium iodide (53 mg, 0.353 mmol) were added to a flask, and the reaction solution was heated to 150 ° C. and reacted for 15 hours. The reaction solution was poured into water, extracted twice with ethyl acetate, and the organic phases were combined, washed with water, washed with saturated saline, spin-dried, and the residue was purified with a silica gel column to obtain 121 mg of product A-15-2, with a yield of 53%.

[0103] The synthesis method of A-15 was based on the synthesis method of A-2 from A-2-3. Preparation of Intermediate A-16:

[0104] [ka]

[0105] The synthesis method of A-16 was based on the synthesis method of A-13 from A-13-1. Preparation of Intermediate A-17:

[0106] [ka]

[0107] Compound A-17-1 (250 mg, 1.76 mmol), dichloromethane (5 mL), p-bromophenylboronic acid (706 mg, 3.52 mmol), copper acetate (320 mg, 1.76 mmol), pyridine (418 mg, 5.28 mmol) and 4A molecular sieves (powder, 500 mg) were added to a flask, and the reaction solution was stirred at room temperature for 2 days under air conditions. Silica gel was added directly to the reaction solution, spin-dried and mixed, and purified with a silica gel column to obtain 367 mg of product A-17-2, with a yield of 70%.

[0108] The synthesis method of A-17 was based on the synthesis method of A-2 from A-2-3. Preparation of Intermediate A-18:

[0109] [ka]

[0110] Compound A-18-1 (500 mg, 2.44 mmol), toluene (10 mL), cyclopropylboronic 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) were added to a flask. The reaction mixture was purged with nitrogen, heated to 100°C and reacted overnight. After cooling the reaction mixture, silica gel was added directly, spin-dried, mixed and purified with a silica gel column to obtain 316 mg of product A-18-2, with a yield of 78%.

[0111] The synthesis method of A-18-3 was based on the synthesis method of A-15-1 from A-2-3.

[0112] The synthesis method of A-18 was carried out in accordance with the synthesis method of A-2 from A-2-2.

[0113] The following compounds were synthesized using the corresponding commercially available starting materials by the methods for preparing the above intermediates.

[0114] [Table 2]

[0115] Preparation of Intermediate A-21:

[0116] [ka]

[0117] Compound A-21-1 (500 mg, 1.91 mmol) and tetrahydrofuran (8 mL) were added to the flask, the reaction solution was replaced with nitrogen, and cooled in an ice-salt bath. Methylmagnesium bromide (1 M tetrahydrofuran solution, 2.3 mL, 2.3 mmol) was added dropwise to the reaction solution. After completion of the addition, the temperature was raised to room temperature and reacted for 1 hour. A saturated aqueous solution of ammonium chloride was added to the reaction solution to quench the reaction, and the mixture was extracted twice with ethyl acetate. The organic phases were combined, washed with water, washed with saturated saline, dried with anhydrous sodium sulfate, suction filtered and spin-dried to obtain 535 mg of product A-21-1, with a yield of 100%. The product was used in the next step as is without further purification.

[0118] The synthesis method of A-21 was based on the synthesis method of A-2 from A-2-3. Preparation of Intermediate A-22:

[0119] [ka]

[0120] For the synthesis of A-2, please refer to the literature Journal of medicinal Chemistry, 2020, vol. 63, #10, 5102-5118.

[0121] The following compounds were synthesized using the corresponding commercially available starting materials by the methods for preparing the above intermediates.

[0122] [Table 3]

[0123] Preparation of Intermediate A-26:

[0124] [ka]

[0125] The synthesis method of A-26-2 was based on the synthesis method of A-18-2 from A-18-1.

[0126] Compound A-26-2 (500 mg, 2.57 mmol), methanol (8 mL) and aqueous sodium hydroxide (1 m, 5.1 mL, 5.1 mmol) were added to a flask, and the reaction solution was allowed to react at room temperature overnight. The reaction solution was diluted with water, adjusted with dilute hydrochloric acid, extracted twice with ethyl acetate, and the combined organic phase was washed with water, washed with saturated saline, dried with anhydrous sodium sulfate, suction filtered and spin-dried to obtain 440 mg of product A-26-3, with a yield of 95%. The product was used directly in the next step without further purification.

[0127] 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 a flask. HATU (633 mg, 1.67 mmol) was added in one portion to the reaction solution, and the mixture was reacted at room temperature overnight. Water was added to the reaction solution to quench the reaction, and the mixture was extracted twice with ethyl acetate. The combined organic phase was washed with water, washed with saturated saline, dried over anhydrous Na2SO4, evaporated in vacuum, and then purified on a silica gel column to obtain 219 mg of product A-26, with a yield of 52%. Preparation of Intermediate A-27:

[0128] [ka]

[0129] Compound A-21-1 (1000 mg, 3.83 mmol), St-butylsulfinamide (511 mg, 4.21 mmol) and 1,4-dioxane (10 mL) were added to a flask, the reaction solution was replaced with nitrogen, and tetraethyl titanate (2184 mg, 9.58 mmol) was added to the reaction solution. The reaction solution was heated to 100 ° C. and stirred for 5 hours. The reaction solution was cooled, quenched by adding water, extracted twice with ethyl acetate, and the organic phases were combined, washed with water, washed with saturated saline, evaporated in vacuum, and then purified with a silica gel column to obtain 882 mg of product A-27-1, with a yield of 63%.

[0130] Compound A-27-1 (882 mg, 2.42 mmol) and tetrahydrofuran (14 mL) were added to a flask, the reaction solution was replaced with nitrogen, and cooled in an ice-salt bath. Methylmagnesium bromide (1 M tetrahydrofuran solution, 2.9 mL, 2.9 mmol) was added dropwise to the reaction solution. After completion of the addition, the temperature was raised to room temperature and reacted for 1 hour. A saturated aqueous solution of ammonium chloride was added to the reaction solution to quench the reaction, and the mixture was extracted twice with ethyl acetate. The organic phase was combined, washed with water, washed with saturated saline, evaporated in vacuum, and then purified with a silica gel column to obtain 630 mg of product A-27-2, with a yield of 68%.

[0131] Compound A-27-2 (630 mg, 1.66 mmol) and methanol (10 mL) were added to a flask, followed by the addition of a solution of hydrogen chloride in 1,4-dioxane (4 M, 6 mL). The reaction solution was reacted at room temperature for 1 hour, then concentrated and dried under reduced pressure. Water was added to the residue, adjusted with an aqueous sodium hydroxide solution, extracted twice with ethyl acetate, and the combined organic phase was washed with water and saturated saline, evaporated in vacuum, and then purified with a silica gel column to obtain 371 mg of product A-27-3, with a yield of 81%.

[0132] The synthesis method of A-27 was based on the synthesis method of A-2 from A-2-3. Preparation of Intermediate A-28:

[0133] [ka]

[0134] The synthesis method of A-28 was carried out in accordance with the synthesis method of A-2 from A-2-3. Preparation of Intermediate A-29:

[0135] [ka]

[0136] Compound A-29-1 (1000 mg, 5.17 mmol) and tetrahydrofuran (10 mL) were added to the flask, the reaction liquid was replaced with nitrogen, and cooled in an ice-water bath. Isopropyl magnesium chloride (1 M tetrahydrofuran solution, 6.2 mL, 6.2 mmol) was added dropwise to the reaction liquid. After the addition was completed, the reaction liquid was warmed to room temperature and stirred for 1 hour, and a solution of p-fluorobenzaldehyde (770 mg, 6.20 mmol) in tetrahydrofuran (4 mL) was added dropwise to the reaction liquid. After the addition was completed, the reaction liquid was stirred at room temperature for 1 hour, and a saturated aqueous ammonium chloride solution was added to the reaction liquid to quench the reaction, and the mixture was extracted twice with ethyl acetate. The organic phase was combined, washed with water, washed with saturated saline, evaporated in vacuum, and then purified with a silica gel column to obtain 612 mg of product A-29-2, with a yield of 50%.

[0137] 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 flask, and the reaction was allowed to react at room temperature for 1 hour, after which TLC showed the reaction was complete. Silica gel was added directly to the reaction, spin-dried, mixed, and purified by silica gel column to obtain 495 mg of product A-29-3, with a yield of 82%.

[0138] The synthesis method of A-29 was based on the synthesis method of A-27 from A-21-1. Preparation of intermediate A-30:

[0139] [ka]

[0140] 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 a flask, and the reaction solution was heated to 70° C. and stirred overnight. The reaction solution was cooled, suction filtered, washed with ethyl acetate, and the filtrate was evaporated in vacuum and then purified with a silica gel column to obtain 742 mg of product A-30-2, with a yield of 97%.

[0141] The synthesis method of A-30 was based on the synthesis method of A-2 from A-2-3. Preparation of Intermediate A-31:

[0142] [ka]

[0143] 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 a flask, 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, and the combined organic phase was washed with water and saturated saline, evaporated in vacuum, and then purified with a silica gel column to obtain 580 mg of product A-31-2, with a yield of 62%.

[0144] The synthesis method of A-31 was based on the synthesis method of A-2 from A-2-3.

[0145] The following compounds were synthesized using the corresponding commercially available starting materials by the methods for preparing the above intermediates.

[0146] [Table 4]

[0147] Preparation of Intermediate A-34:

[0148] [ka]

[0149] 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 a flask. HBTU (1449 mg, 3.82 mmol) was added to the reaction solution all at once while stirring. The reaction solution was stirred at room temperature overnight, poured into water, extracted four times with ethyl acetate, the organic phase was combined, washed with saturated saline, evaporated in vacuum, and then purified with a silica gel column to obtain 600 mg of product A-34-2, with a yield of 96%.

[0150] Compound p-bromoiodobenzene (876 mg, 3.10 mmol) and tetrahydrofuran (10 mL) were added to the flask, the reaction solution was replaced with nitrogen, and cooled to -70 ° C. in a dry ice / ethanol bath. 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 completion of the dropwise addition, the reaction solution was slowly warmed to room temperature and reacted for 1 hour. The reaction solution was quenched by adding water to the reaction solution, extracted twice with ethyl acetate, the organic phase was combined and washed with saturated saline, evaporated in vacuum, and then purified with a silica gel column to obtain 360 mg of product A-34-3, with a yield of 41%.

[0151] The synthesis method of A-34 was carried out in accordance with the synthesis method of A-2 from A-2-3. Preparation of Intermediate A-35:

[0152] [ka]

[0153] The synthesis method of A-35-2 was based on the synthesis method of A-34-3 from A-34-2. The synthesis method of A-35-3 was based on the synthesis method of A-29-3 from A-29-2. The synthesis method of A-35 was carried out in accordance with the synthesis method of A-2 from A-2-3. The following compounds were synthesized using the corresponding commercially available starting materials by the methods for preparing the above intermediates.

[0154] [Table 5]

[0155] Preparation of intermediate A-40:

[0156] [ka]

[0157] 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 a flask. The reaction solution was heated to 80° C. and reacted for 5 hours. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, and the combined organic phase was washed with water, washed with saturated saline, dried over anhydrous sodium sulfate, evaporated in vacuum, and then purified on a preparative silica gel plate to obtain 62 mg of product A-40, with a yield of 60%.

[0158] The following compounds were synthesized using the corresponding commercially available starting materials by the methods for preparing the above intermediates.

[0159] [Table 6]

[0160] Preparation of Intermediate A-48:

[0161] [ka]

[0162] Compound A-15-1 (200 mg, 0.706 mmol), DMF (4 mL) and sodium difluorochloroacetate (215 mg, 1.41 mmol) were added to a flask, the reaction solution was purged with nitrogen, and the mixture was heated to 100 ° C. and reacted for 6 hours. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, and the combined organic phase was washed with water and saturated saline, and evaporated in vacuum. The product was then purified with a silica gel column to obtain 128 mg of product A-48-1, with a yield of 54%.

[0163] The synthesis method of A-48 was carried out in accordance with the synthesis method of A-2 from A-2-3. The following compounds were synthesized using the corresponding commercially available starting materials by the methods for preparing the above intermediates.

[0164] [Table 7]

[0165] Preparation of intermediate A-54:

[0166] [ka]

[0167] 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 a flask, and the reaction solution was heated to 100 ° C. and reacted for 4 hours. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, and the combined organic phase was washed with water and saturated saline, and then evaporated in vacuum. The product A-54-2 (511 mg) was obtained by purification on a silica gel column, with a yield of 46%.

[0168] The synthesis method of A-54 was based on the synthesis method of A-2 from A-2-3. Preparation of Intermediate A-55:

[0169] [ka]

[0170] Compound A-55-1 (1.00 g, 7.35 mmol), Pd / C (10%, 200 mg) and methanol (25 mL) were added to the flask, and the reaction mixture was substituted with hydrogen, followed by stirring overnight under hydrogen pressure (balloon). The reaction mixture was filtered by suction, and the filtrate was directly spin-dried to obtain 1.00 g of product A-55-2, with a yield of 99%. The product was used in the next step without purification.

[0171] Compound A-55-2 (800 mg, 5.79 mmol) and tetrahydrofuran (10 mL) were added to a flask, the reaction solution was replaced with nitrogen, and cooled to -70 ° C. in a dry ice / ethanol bath. 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 boronate (723 mg, 6.95 mmol) in tetrahydrofuran (3 mL) was added dropwise to the reaction solution. After completion of the addition, the reaction solution was slowly warmed to room temperature and reacted for 30 minutes. The reaction was quenched with dilute hydrochloric acid, extracted twice with ethyl acetate, the organic phases were combined and washed with saturated saline, evaporated in vacuum, and then purified with a silica gel column to obtain 430 mg of product A-55-3, with a yield of 41%.

[0172] The synthesis method of A-55 was carried out in accordance with the synthesis method of A-2 from A-2-1. Preparation of Intermediate A-56:

[0173] [ka]

[0174] 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 a flask, and the reaction solution was heated to 100° C. and reacted for 4 hours. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, and the combined organic phase was washed with water and saturated saline, and then evaporated in vacuum. The product A-56-2 was obtained in an amount of 320 mg and purified by a silica gel column with a yield of 59%.

[0175] The synthesis method of A-56 was based on the synthesis method of A-55 from A-55-2. The following compounds were synthesized using the corresponding commercially available starting materials by the methods for preparing the above intermediates.

[0176] [Table 8]

[0177] Preparation of intermediate A-60:

[0178] [ka]

[0179] 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 a 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, and the combined organic phase was washed with water and saturated saline, and then evaporated in vacuum. The product A-60-2 (830 mg) was obtained by purification on a silica gel column, with a yield of 82%.

[0180] The synthesis method of A-60 was based on the synthesis method of A-2 from A-2-3. The following compounds were synthesized using the corresponding commercially available starting materials by the methods for preparing the above intermediates.

[0181] [Table 9]

[0182] Preparation of Intermediate A-64:

[0183] [ka]

[0184] Compound A-64-1 (1.192 g, 8.05 mmol), bromobenzene (10.1 g, 64.3 mmol) and aluminum trichloride (2.15 g, 16.1 mmol) were added to a flask, the reaction liquid was replaced with nitrogen, and the mixture was heated to 90 ° C. and reacted for 3 hours. The reaction liquid was cooled, poured into dilute hydrochloric acid, extracted three times with dichloromethane, and the organic phases were combined. The organic phase was extracted three times with aqueous sodium carbonate solution, the aqueous phase was adjusted to pH 3 with dilute hydrochloric acid, the precipitated solid was suction filtered, washed with water, the filter cake was collected, and dried in vacuum to obtain 2.0 g of product A-64-2, with a yield of 81%. The product was used directly in the next step without further purification.

[0185] The synthesis method of A-64-3 was based on the synthesis method of A-2 from A-2-3.

[0186] 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 a flask. HBTU (324 mg, 0.85 mmol) was added in one portion to the reaction solution while stirring. The reaction solution was stirred at room temperature overnight, poured into water, extracted three times with ethyl acetate, the combined organic phase was washed with saturated saline, evaporated in vacuum, and then purified on a silica gel preparative plate to give 70 mg of product A-64, with a yield of 35%.

[0187] The following compounds were synthesized using the corresponding commercially available starting materials by the methods for preparing the above intermediates.

[0188] [Table 10]

[0189] Preparation of Intermediate A-68:

[0190] [ka]

[0191] 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 a flask, and the reaction solution was heated to 180 ° C. and reacted for 8 hours. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, and the combined organic phase was washed with water, washed with saturated saline, dried with anhydrous sodium sulfate, evaporated in vacuum, and then purified with a silica gel separation plate to obtain 60 mg of product A-68-2, with a yield of 15%.

[0192] The synthesis method of A-68 was based on the synthesis method of A-2 from A-2-3.

[0193] Preparation of Intermediate A-69:

[0194] [ka]

[0195] The synthesis method of A-69 was carried out in accordance with the synthesis method of A-1 from A-1-1 and A-1-2. Preparation of intermediate A-70:

[0196] [ka]

[0197] 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 a flask, and the reaction solution was stirred at room temperature overnight. The reaction solution was poured into water, extracted twice with ethyl acetate, and the organic phases were combined and evaporated in vacuum, and then purified on a silica gel column to obtain 190 mg of product A-70-1, with a yield of 100%.

[0198] The synthesis method of A-70 was based on the synthesis method of A-2 from A-2-3. Preparation of Intermediate A-71:

[0199] [ka]

[0200] The synthesis method of A-71 was based on the synthesis method of A-54 from A-54-1. Preparation of Intermediate A-72:

[0201] [ka]

[0202] 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) were added to a flask. The reaction solution was heated to 60° C. and reacted overnight. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, and the combined organic phase was washed with water and saturated saline, evaporated in vacuum, and then purified with a silica gel column to obtain 1.56 g of product A-72, with a yield of 49%.

[0203] Preparation of intermediate A-73:

[0204] [ka]

[0205] 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 the mixture was heated to 90° C. and reacted overnight. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, and the combined organic phase was washed with saturated saline, dried over anhydrous sodium sulfate, evaporated in vacuum, and then purified with a silica gel preparative plate to obtain 270 mg of product A-73-1, with a yield of 83%.

[0206] The synthesis method of A-73 was carried out in accordance with the synthesis method of A-2 from A-2-3. Preparation of intermediate A-74:

[0207] [ka]

[0208] 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 flask, the reaction was purged with nitrogen and cooled to -70 ° C in a dry ice / ethanol bath. n-Butyl lithium (2.5 m, 3.3 mL, 8.30 mmol) was added dropwise to the reaction, and the mixture was stirred and kept warm for 2 hours. Dry ice was carefully added to the reaction, and the reaction was slowly warmed to room temperature. The reaction was quenched with dilute hydrochloric acid, extracted three times with dichloromethane, and the combined organic phase was washed with saturated saline, dried over anhydrous sodium sulfate and evaporated in vacuum to give 1.00 g of product A-74-2, with a yield of 74%. The product was used directly in the next step without further purification.

[0209] Compound A-74-2 (800 mg, 4.70 mmol) and dichloromethane (8 mL) were added to the flask, the reaction liquid was replaced with nitrogen, and cooled in an ice-water bath. A solution of boron tribromide in dichloromethane (17%, 27.7 g, 18.8 mmol) was added dropwise to the reaction liquid. After the addition was completed, the reaction liquid was warmed to room temperature and reacted for 30 minutes, cooled again in an ice-water bath, and methanol was slowly added dropwise to quench the reaction. The reaction liquid was directly evaporated in vacuum, and then purified with a silica gel column to obtain 560 mg of product A-74-3 in a yield of 76%.

[0210] Compound A-74-3 (560 mg, 3.59 mmol), DMF (5 mL), alcohol solution of methylamine (30%, 557 mg, 5.38 mmol), DIEA (1392 mg, 10.8 mmol) and HATU (1775 mg, 4.67 mmol) were added to a flask. The reaction was stirred at room temperature overnight. The reaction was poured into water and extracted six times with ethyl acetate, the organic phases were combined and evaporated in vacuum, and then purified on a silica gel column to give 254 mg of product A-74-4, with a yield of 42%.

[0211] The synthesis method of A-74 was based on the synthesis method of A-17 from A-17-1. Preparation of Intermediate A-75:

[0212] [ka]

[0213] The synthesis method of A-75-1 was based on the synthesis method of A-34-2 from A-34-1.

[0214] Compound A-75-1 (500 mg, 1.91 mmol) and tetrahydrofuran (8 mL) were added to the flask, the reaction liquid 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 liquid, and after the addition was completed, the reaction liquid was slowly warmed to room temperature and stirred for 1 hour. The reaction was quenched with dilute hydrochloric acid, extracted twice with ethyl acetate, and the combined organic phase was washed with water, washed with saturated saline, dried over anhydrous sodium sulfate, and evaporated in vacuum to give 538 mg of product A-75-2, with a yield of 100%. The product was used directly in the next step without further purification.

[0215] The synthesis method of A-75 was carried out in accordance with the synthesis method of A-73 from A-21-1. Preparation of Intermediate A-76:

[0216] [ka]

[0217] 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 a 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, and the combined organic phase was washed with water and saturated saline, evaporated in vacuum, and then purified with a silica gel column to obtain 280 mg of product A-76-2 in a yield of 76%.

[0218] Compound A-76-2 (200 mg, 0.597 mmol), sodium methanol (161 mg, 2.98 mmol) and NMP (2 mL) were added to a 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, and the combined organic phase was washed with water and saturated saline, and then evaporated in vacuum. The product A-76-3 (116 mg) was obtained by purification on a silica gel column, with a yield of 56%.

[0219] The synthesis method of A-76 was based on the synthesis method of A-2 from A-2-3. Preparation of Intermediate A-77:

[0220] [ka]

[0221] The synthesis method of A-77-3 was based on the synthesis method of A-34-3 from A-34-1.

[0222] The synthesis method of A-77 was based on the synthesis method of A-73 from A-21-1. Preparation of Intermediate A-78:

[0223] [ka]

[0224] The synthesis method of A-78-2 was based on the synthesis method of A-34-2 from A-34-1.

[0225] 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 a flask, and the reaction solution was replaced with nitrogen, heated to 110 ° C and reacted overnight. The reaction solution was cooled, poured into water, extracted twice with ethyl acetate, and the combined organic phase was washed with saturated saline, evaporated in vacuum, and then purified with a silica gel column to obtain 473 mg of product A-78-3, with a yield of 100%.

[0226] The synthesis method of A-78-4 was based on the synthesis method of A-34-3 from A-34-2. The synthesis method of A-78 was based on the synthesis method of A-73 from A-21-1. The following compounds were synthesized using the corresponding commercially available starting materials by the methods for preparing the above intermediates.

[0227] [Table 11]

[0228] Preparation of Intermediate A-82:

[0229] [ka]

[0230] Dichloromethane (5mL) was added to the flask, substituted with nitrogen, and cooled to -40°C in a dry ice / acetonitrile bath. Titanium tetrachloride (1453mg, 7.66mmol) was added, and then a toluene solution of dimethylzinc (1m, 7.7mL, 7.7mmol) was slowly added dropwise to the reaction solution, and the mixture was allowed to react while keeping warm for 30 minutes. A dichloromethane (3mL) solution of compound A-21-1 (500mg, 1.91mmol) was added dropwise to the reaction solution, and the mixture was allowed to react while keeping warm for 1 hour, and then the mixture was slowly heated to room temperature and stirred overnight. The reaction was quenched by adding water to the reaction solution, and the reaction was extracted twice with dichloromethane. The organic phase was combined and washed with water, evaporated in vacuum, and then purified with a silica gel column to obtain 326mg of product A-82-1, with a yield of 62%.

[0231] The synthesis method of A-82 was based on the synthesis method of A-2 from A-2-3. The following compounds were synthesized using the corresponding commercially available starting materials by the methods for preparing the above intermediates.

[0232] [Table 12] JPEG0007660800000084.jpg255167JPEG0007660800000085.jpg99170

[0233] Preparation of Intermediate A-101:

[0234] [ka]

[0235] Add p-bromoiodobenzene (1.71g, 6.03mmol) and tetrahydrofuran (15mL) to a flask, replace with nitrogen, and cool to -70℃ in a dry ice / ethanol bath. Then, slowly add n-butyllithium (2.5m, 2.4mL, 6.03mmol) and react for 30 minutes while keeping warm. Weigh out A-101-1 (1.00g, 5.74mmol) and dissolve it in tetrahydrofuran (5mL) and dropwise add it to the reaction solution. After 10 minutes, the reaction solution was slowly warmed to room temperature. Add water to the reaction solution to quench the reaction, extract twice with ethyl acetate, combine the organic phase, wash with water, evaporate in vacuum, and then purify with a silica gel column to obtain 1.4g of product A-101-2, with a yield of 73%.

[0236] The synthesis method of A-101 was based on the synthesis method of A-2 from A-2-3. Preparation of Intermediate A-102:

[0237] [ka]

[0238] A flask was charged with A-101-2 (550 mg, 1.66 mmol) and dichloromethane (6 mL), and the mixture was replaced with nitrogen. DAST (402 mg, 2.49 mmol) was added in an ice bath and the mixture was allowed to react for 2 hours. The reaction mixture was quenched by adding an aqueous solution of sodium bicarbonate, extracted twice with dichloromethane, and the organic phases were combined, dried over anhydrous sodium sulfate, evaporated in vacuum, and purified on a silica gel preparative plate to obtain 410 mg of product A-102, with a yield of 74%.

[0239] The synthesis method of A-102 was based on the synthesis method of A-2 from A-2-3. Preparation of intermediate A-103:

[0240] [ka]

[0241] A-103-1 (500 mg, 1.50 mmol) and an aqueous solution of hydrobromic acid (5 mL) were added to a flask, cooled in an ice bath, sodium nitrite (645 mg, 9.35 mmol) was added, and the mixture was allowed to react for 20 minutes while keeping the temperature constant. A hydrobromic acid aqueous solution (5 mL) of CuBr (2.69 g, 18.75 mmol) was added to the reaction solution, and the reaction solution was slowly warmed to room temperature and allowed to react for 3 hours. The reaction solution was diluted with water, extracted twice with ethyl acetate, and the organic phases were combined and washed with water, evaporated in vacuum, and then purified with a silica gel column to obtain 620 mg of product A-103-2, with a yield of 89%.

[0242] A flask was charged with A-103-2 (200 mg, 0.43 mmol) and tetrahydrofuran (5 mL), purged with nitrogen, cooled to -70 °C in a dry ice / ethanol bath, n-butyllithium (2.5 m, 0.17 mL, 0.43 mmol) was slowly added dropwise, and the reaction was allowed to proceed for 1 hour after the addition. The reaction was quenched with dilute hydrochloric acid, extracted twice with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, and evaporated in vacuum to give 170 mg crude product A-103-3, which was used in the next step without purification.

[0243] The synthesis method of A-103 was based on the synthesis method of A-2 from A-2-3. Preparation of Intermediate B-1:

[0244] [ka]

[0245] 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 flask, the reaction solution was purged with nitrogen, TBDPSCl (5.30 g, 19.3 mmol) was added in an ice-water bath, after the dropwise addition was completed, the ice was removed, and the reaction mixture was stirred at room temperature for 16 hours, after which TLC showed that the reaction was complete. The reaction solution was poured into water, and the reaction solution was extracted twice with ethyl acetate, combined, washed twice with water, then washed with saturated NaCl solution, and finally dried over anhydrous Na2SO4 and evaporated directly in vacuum to give 6.68 g of product B-1-2, which was used directly in the next step without purification.

[0246] A tetrahydrofuran solution (35 mL) of B-1-2 (6.68 g, 18.9 mmol) was added to the flask, the reaction liquid was replaced with nitrogen, and 9-BBN (0.5 m, 91 mL) was added dropwise under an ice-water bath. After the addition was completed, the reaction mixture was stirred at room temperature for 16 hours, and TLC showed that the reaction of the raw materials was complete. The reaction liquid was cooled again in an ice-water bath, and 10% NaOH solution (24 mL) and 30% H2O2 solution (12 mL) were slowly added, and stirring was continued for 1 hour, and TLC showed that the reaction of the intermediate was complete. The reaction liquid was poured into water, and the mixture was extracted twice with ethyl acetate, washed twice with water, washed with saturated NaCl solution, and mixed by directly adding silica gel, and then purified by silica gel column to obtain 6.49 g of product B-1-3, and the yield of the two steps was 100%.

[0247] Compound B-1-3 (6.49 g, 17.5 mmol) in dichloromethane (50 mL) was added to the flask, the reaction liquid was replaced with nitrogen, Dess-Martin oxidant (11.14 g, 26.3 mmol) was added to the flask in an ice-water bath, and after stirring for 1.5 hours, TLC showed that the reaction was complete. Silica gel was added directly and mixed, then purified by silica gel column to obtain 6.45 g of product B-1-4, with a yield of 100%.

[0248] 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) were added to a flask, substituted with nitrogen, and cooled in an ice-water bath. t-BuOK (3.83 g, 34.1 mmol) was added to the reaction solution, stirred for 30 minutes, then slowly warmed to room temperature and continued stirring for 1.5 hours. The raw material reaction was complete by TLC. The reaction solution was poured into a saturated NH4Cl solution (350 mL), extracted twice with ethyl acetate, combined, added silica gel, mixed, and then purified with a silica gel column to obtain 2.27 g of product B-1-5 (2 spots were shown on TLC), with a yield of 34%.

[0249] A dichloromethane solution (35 mL) of B-1-5 (2.27 g, 5.97 mmol) was added to the flask, the reaction liquid was purged with nitrogen, cooled in a dry ice-ethanol bath, Dibal-H (1 M, 9 mL) was slowly added dropwise to the reaction liquid, and stirred at this temperature for 1.5 hours. After TLC showed that the reaction was complete, the reaction liquid was added to cold dilute hydrochloric acid (1 M, 10 mL) and stirred until no bubbles remained, then extracted twice with DCM, the combined organic phase was washed with saturated NaCl solution, and finally dried over anhydrous Na2SO4, and the reaction liquid was directly evaporated in vacuum to give 2.22 g of product B-1-6. The yield was 97%.

[0250] 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 flask, then KMnO4 (3.67 g, 23.2 mmol) was added in portions, and the reaction solution was stirred at room temperature for 30 minutes. After TLC showed that the raw materials were completely reacted, the reaction solution was adjusted by adding dilute hydrochloric acid, and further NaHSO3 solution was added until the reaction solution became colorless, and the reaction solution was extracted twice with ethyl acetate, and the organic phases were combined and washed with saturated NaCl solution, and finally dried with anhydrous Na2SO4, and the reaction solution was directly evaporated in vacuum to obtain 1.89 g of product B-1-7. The yield was 82%.

[0251] 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) were added to a flask, substituted with nitrogen, cooled in an ice-water bath, and HATU (2.16 g, 5.69 mmol) and DIEA (3.06 g, 23.7 mmol) were added to the reaction solution. The reaction solution was stirred for 30 minutes, then the ice was removed and stirring was continued at room temperature for 30 minutes. The raw material reaction was complete according to TLC. The reaction solution was poured into water, extracted twice with ethyl acetate, combined, washed twice with water, and further washed with saturated NaCl solution, mixed with silica gel, and then purified with a silica gel column to obtain 1.94 g of product B-1-8 (2 spots were shown on TLC). The yield was 78%.

[0252] Compound B-1-8 (790 mg, 1.51 mmol), DMF (0.8 mL) and ethyl acetate (8 mL) were added to a flask, and the atmosphere was replaced with nitrogen. 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 hour. The raw material reaction was complete according to TLC. The reaction solution was poured into NaHCO3 solution (4.5 g NaHCO3 / 30 mL h2O) to quench it, and the mixture was extracted twice with ethyl acetate, washed twice with water, and then washed with saturated NaCl solution, and finally dried over anhydrous Na2SO4 and evaporated directly in vacuum to obtain 680 mg of product B-1-9 (2 spots were shown on TLC). The yield was 89.0%.

[0253] A DMF solution (7 mL) of B-1-9 (680 mg, 1.34 mmol) was added to the flask, the reaction solution was purged with nitrogen, NBS (287 mg, 1.61 mmol) was added under ice-water bath, and after stirring for 40 minutes, the reaction was complete by TLC. The reaction solution was quenched by pouring into water, and the two extractions with ethyl acetate were combined, washed twice with water, and further washed with saturated NaCl solution, mixed with silica gel, and then purified on a silica gel column to obtain 298 mg of product B-1-10-A (a spot with a low polarity appears first on TLC) and 339 mg of product B-1-10-B (a spot with a high polarity appears later on TLC). The total yield was 81%.

[0254] Compound B-1-10-A (298 mg, 0.509 mmol), aqueous ammonia (6 mL), and n-butanol solution (3 mL) were added to a high-pressure digestion vessel, and the flask was sealed and heated to 95° C. and stirred for 16 hours. The reaction solution was cooled and spin-dried in vacuum, and then purified on a silica gel column to obtain 184 mg of product B-1-A, with a yield of 64%.

[0255] According to the synthesis method of B-1-A, B-1-10-B was reacted with ammonia water to prepare B-1-B. Preparation of Intermediate B-2:

[0256] [ka]

[0257] Weigh out NaH (8.56 g, 357 mmol), suspend in anhydrous THF (80 mL), heat to 40-45 ° C, add dropwise a THF solution of compound B-2-1 (22.85 g, 149 mmol), and after completion of the dropwise addition, keep at this temperature while stirring and reacting for 15 minutes. Add dropwise a THF solution of ethyl acrylate, and continue the reaction for 15 minutes after the addition. After cooling the reaction solution to room temperature, add it to ice water, adjust the pH to 3 with concentrated HCl, add ethyl acetate and extract twice, combine the organic phase, dry with 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%).

[0258] Compound B-2-2 (37.6 g, 120 mmol), 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 hours. 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. Purification by column chromatography gave compound B-2-3 as a colorless oil (21.6 g, 75%).

[0259] Compound B-2-3 (21.6 g, 89.2 mmol), ethylene glycol (6.64 g, 107 mmol) and p-toluenesulfonic acid hydrate (169 mg, 0.89 mmol) were weighed and added to toluene (180 mL), and the mixture was refluxed for 4 hours at 120° C. to separate water. The reaction solution was cooled, added to a saturated NaHCO3 aqueous 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 pale yellow liquid (23.7 g, 93%).

[0260] LAH (6.47g, 166mmol) was weighed and placed in a three-neck flask, anhydrous THF (150mL) was added, the mixture was replaced with nitrogen, and a solution of compound B-2-4 (23.7g, 82.8mmol) in THF (100mL) was added dropwise under ice-salt bath cooling. After the addition was completed, the mixture was slowly heated to room temperature and reacted for 5 hours. H2O / THF (1:1, 30mL) was slowly added dropwise to the reaction in an ice-water bath, and 5N aqueous sodium hydroxide solution (8mL) was added and stirred at room temperature overnight. The reaction solution was diluted by adding DCM / MeOH (5:1, 250mL) to the flask, filtered, and rinsed with DCM / MeOH (5:1). 50g silica gel was added to the filtrate, stirred for 15 minutes, filtered, and rinsed. The filtrate was concentrated under reduced pressure to obtain compound B-2-5 (16.7g, 99%).

[0261] Compound B-2-5 (16.7 g, 82.6 mmol) was weighed and placed in a flask, pyridine (100 mL) was added, TsCl (34.6 g, 182 mmol) was added under ice water bath, and the mixture was stirred at room temperature overnight. The reaction solution was diluted with ethyl acetate, washed with 10% citric acid solution and saturated sodium chloride, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained crude product was slurried with ethanol to obtain compound B-2-6 as a white solid (35 g, 83%).

[0262] Compound B-2-6 (35 g, 68.5 mmol) was weighed and placed in a flask, and 1N HCl solution (260 mL) and THF (300 mL) were added and reacted at 80° C. for 5 hours. The reaction solution was extracted with ethyl acetate, and the organic phase was washed with water, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound B-2-7 (26.2 g, 82%) was obtained by purifying it by column chromatography.

[0263] Compound 1,3-dithiane (2.1 g, 17.4 mmol) and anhydrous THF (40 mL) were added to a flask, and the mixture was replaced with nitrogen. Under cooling in a dry ice-ethanol bath, n-butyllithium (2.5 M, 8.5 mL) was added dropwise. After completion of the addition, the mixture was heated to 0°C and reacted for 1 hour. After cooling in a dry ice-ethanol bath, a THF solution of compound B-2-7 (6.5 g, 13.9 mmol) was added dropwise. After completion of the addition, the mixture was heated to room temperature and reacted for 1 hour. The reaction solution was quenched by adding it 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-2-8 (6.77 g, 83%) was obtained by purifying it by column chromatography.

[0264] Compound B-2-8 (6.77 g, 11.5 mmol), NaOH (1.38 g, 34.6 mmol), and THF (170 mL) were added to a flask and refluxed at 70 ° C. overnight. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound B-2-9 (3.7 g, 77%) was obtained by purification using column chromatography.

[0265] Compound B-2-9 (3.7 g, 8.92 mmol), acetonitrile (50 mL) and water (12.5 mL) were added to a flask, and NBS (5.56 g, 31.2 mmol) was added under ice-water bath. After the addition, the mixture was transferred to room temperature and reacted for 3 hours. The reaction solution was added to a saturated NaHCO3 solution, extracted with ethyl acetate, and the organic phases were combined and back-extracted with a saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product of compound B-2-10. This crude product was not purified.

[0266] The crude product of compound B-2-10 obtained above was dissolved in ethanol, and NaBH4 (508 mg, 13.4 mmol) was added under ice water bath, and the mixture was transferred to room temperature and reacted for 1 hour. 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, 91% yield for two steps) was obtained by purifying it by column chromatography.

[0267] 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) were added to a flask, and TBDPSCl (2.59 g, 9.41 mmol) was added and reacted at room temperature for 0.5 hours. The reaction solution was added to water, extracted with ethyl acetate, and the organic phases were combined and back-extracted with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound B-2-12 (4.0 g, 90%) was obtained by purification using column chromatography.

[0268] Compound B-2-12 (4.0 g, 7.08 mmol) and methanol (120 mL) were added to a flask, and Mg (1.89 g, 77.9 mmol) was added while stirring at room temperature. After 30 minutes, the reaction became highly 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 purification by column chromatography.

[0269] Compound B-2-13 (2.4 g, 5.85 mmol) and DMF (30 mL) were added to a flask, and PDC (6.6 g, 17.6 mmol) was added under ice water bath. After the addition, the mixture was transferred to room temperature and reacted for 2 hours. Ethyl acetate was added to the flask to dilute the reaction solution, which was then extracted with water. The organic phases were combined and back-extracted with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound B-2-14 (1.99 g, 80%) was obtained by purification using column chromatography.

[0270] 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) were added to the flask, and HBTU (2.13 g, 5.62 mmol) and DIEA (2.42 g, 18.8 mmol) were added to the reaction solution, and the mixture was reacted at room temperature for 1 hour. The reaction solution was added to water, extracted twice with ethyl acetate, and the organic phases were combined and back-extracted with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound B-2-15 (1.99 g, 77%) was obtained by purification using column chromatography.

[0271] Compound B-2-15 (1.59 g, 2.89 mmol) was added to a flask, and DCM (30 mL) was added to dissolve the compound. 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, and the mixture was then transferred to room temperature and reacted for 4 hours. The reaction solution was added to a saturated NaHCO3 solution, extracted with ethyl acetate, and the organic phases were combined and back-extracted with a saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product of compound B-2-16. This crude product was not purified.

[0272] The crude product of compound B-2-16 obtained above was dissolved in DMF (8 mL), NBS (566 mg, 3.18 mmol) was added, and the mixture was reacted at room temperature for 0.5 hours. The reaction solution was added to NaHCO3 solution, extracted with ethyl acetate, and the organic phases were combined and back-extracted with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound B-2-17 (1.43 g, 81% yield for two steps) was obtained by purification using column chromatography.

[0273] Compound B-2-17 (1.43 g, 2.34 mmol), aqueous ammonia (20 mL), and n-butanol (8 mL) were added to a high-pressure digestion tank, and the reaction system was heated to 95° C. and stirred for 16 hours. The reaction solution was spin-dried in vacuum and purified by column chromatography to obtain compound B-2 (1.2 g, 87%). Preparation of intermediate B-3:

[0274] [ka]

[0275] B-3-1 was prepared with reference to the method in the literature (Angew. Chem. Int. Ed. 2020, 59, 7161-7167).

[0276] Compound B-3-1 (1.25 g, 6.71 mmol), imidazole (548 mg, 8.06 mmol) and DMF (12 mL) were added to a flask, and TBDPSCl (1.94 g, 7.05 mmol) was added and reacted at room temperature overnight. The reaction solution was added to water, extracted twice with ethyl acetate, and the combined organic phase was 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.

[0277] 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, adjusted with dilute hydrochloric acid, extracted twice with ethyl acetate, and the combined organic phase was 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, 95%). This compound was not further purified.

[0278] For the method of producing B-3 from B-3-3, reference was made to the method of producing B-2 from B-2-14. Preparation of Intermediate B-4:

[0279] [ka]

[0280] 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 a flask, and the reaction solution was replaced with nitrogen and heated to 60°C. DIAD (194 mg, 0.958 mmol) was added dropwise to the reaction solution and allowed to react overnight while keeping it warm. The reaction solution was cooled, concentrated and dried under reduced pressure, and purified with a silica gel column to obtain compounds B-4-A (52 mg, low polarity spot) and B-4-B (140 mg, high polarity spot, containing impurity triphenylphosphine oxide). The total yield was 82%. Preparation of Intermediate B-5:

[0281] [ka]

[0282] Compound B-1-3 (500 mg, 1.35 mmol), TEA (273 mg, 2.70 mmol), DCM (5 mL) and paratoluenesulfonyl chloride (309 mg, 1.62 mmol) were added to the flask, and the reaction solution was stirred at room temperature for 2 hours, after which TLC showed that the reaction had almost ceased. DMAP (198 mg, 1.62 mmol) was added to the reaction solution and reacted overnight. The reaction solution was added to water, adjusted with dilute hydrochloric acid, extracted twice with ethyl acetate, and the organic phases were combined and washed with water, washed with saturated NaCl solution, concentrated and dried under reduced pressure, and purified with a silica gel column to obtain compound B-5-1 (550 mg, 78%).

[0283] 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 a flask, and the reaction was heated to 100 °C and stirred overnight. The reaction was cooled, added to water, extracted twice with ethyl acetate, and the combined organic phase was washed with water and saturated NaCl solution, concentrated and dried under reduced pressure, and purified on a silica gel preparative plate to obtain compound B-5-2 (60 mg, 27%).

[0284] Compound B-5-2 (60 mg, 0.103 mmol), aqueous ammonia (2 mL), and n-butanol (1 mL) were added to a sealed tube, and the reaction system was heated to 100° C. and stirred overnight. The reaction solution was cooled, spun dry in vacuum, and purified on a silica gel preparative plate to give compound B-5 (38 mg, 66%). Preparation of Intermediate B-6:

[0285] [ka]

[0286] 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 flask, and the reaction was stirred at room temperature for 4 h. The reaction was then directly purified on a silica gel preparative plate to give compound B-6-1 (108 mg, 91%).

[0287] Compound B-6-1 was oxidized with PDC (see the synthesis of B-2-13 to B-2-14) to prepare B-6-2.

[0288] 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 a flask, and the reaction solution was stirred at room temperature overnight. The reaction solution was added to water, extracted twice with ethyl acetate, and the combined organic phase was washed with water, washed with saturated NaCl solution, dried over anhydrous sodium sulfate, suction filtered, concentrated and dried under reduced pressure, and purified on a silica gel preparative plate to obtain compound B-6-3 (80 mg, 85%).

[0289] Compound B-6-3 (80 mg, 0.21 mmol) and THF (2 mL) were added to a flask, which was then replaced with nitrogen and cooled in an ice-water bath. Tetraisopropyl orthotitanate (28 mg, 0.10 mmol) was added to the reaction solution, and ethylmagnesium bromide (0.6 mL, 0.6 mmol, 1 m) was slowly added dropwise. After the addition was completed, the reaction solution was warmed to room temperature and stirred overnight. The reaction solution was poured into an aqueous solution of ammonium chloride to quench the reaction, extracted twice with ethyl acetate, and the organic phases were combined and washed with saturated saline, concentrated and dried under reduced pressure, and purified on a silica gel separation plate to obtain 22 mg of compound B-6-4, with a yield of 28%.

[0290] For the method of producing B-6 from B-6-4, refer to the method of producing B-2 from B-2-17. Preparation of Intermediate B-7:

[0291] [ka]

[0292] Compound B-6-3 (100 mg, 0.267 mmol) and THF (2 mL) were added to a flask, which was then purged 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 was completed, the reaction solution was warmed to room temperature and stirred overnight. The reaction solution was quenched by pouring it into an aqueous solution of ammonium chloride, extracted twice with ethyl acetate, and the organic phases were combined and washed with saturated saline, concentrated and dried under reduced pressure, and purified on a silica gel separation plate to obtain 65 mg of compound B-7-1, with a yield of 65%.

[0293] For the method of producing B-7 from B-7-1, refer to the method of producing B-2 from B-2-17. Preparation of Intermediate B-8:

[0294] [ka]

[0295] Compound B-8-1 (CAS: 652-67-5, 1.00 g, 6.84 mmol), imidazole (559 mg, 8.21 mmol) and DMF (15 mL) were added to a flask, and TBDPSCl (1.88 g, 6.84 mmol) was further added and reacted at room temperature overnight. The reaction solution was poured into water, extracted twice with ethyl acetate, and the combined organic phase was washed with water and saturated NaCl solution, concentrated under reduced pressure, and then purified with a silica gel column to obtain compound B-8-2 (1.63 g, 62%).

[0296] For the method of producing B-8 from B-8-2, reference was made to the method of producing B-1-A(B) from B-1-3. Preparation of Intermediate B-9:

[0297] [ka]

[0298] Compound B-9-1 (8.0 g, 76.1 mmol), dioxane (120 mL) and Raney Ni (approximately 1 g) were added to the flask, and the reaction solution was substituted with hydrogen, heated to 90 ° C under hydrogen bag pressure, and reacted with stirring overnight, and TLC showed that the reaction of the raw materials was almost complete. The reaction solution was cooled, suction filtered, and the filtrate was spin-dried under reduced pressure to obtain 8.3 g of product B-9-2, with a yield of 100%. The product was used in the next step as it was without purification.

[0299] B-9-2 was condensed with B-1-7, ring-closed, and brominated to prepare B-9. For the specific method, refer to the method for producing B-1-10-A(B) from B-1-7. Preparation of Intermediate B-10:

[0300] [ka]

[0301] Compound B-1-9 (200 mg, 0.395 mmol) and tetrahydrofuran (3 mL) were added to the flask, the reaction solution was replaced with nitrogen, and the mixture was cooled to -70 ° C. in a dry ice / ethanol bath. n-Butyl lithium (2.5 m, 0.19 mL, 0.474 mmol) was added dropwise to the reaction solution. After the addition was completed, the mixture was allowed to react while keeping it warm for 30 minutes. Iodomethane (112 mg, 0.790 mmol) was added dropwise to the reaction solution, and after the addition was completed, the reaction solution was slowly warmed to room temperature. The reaction was quenched by adding an aqueous solution of ammonium chloride, extracted twice with ethyl acetate, and the organic phases were combined and concentrated and dried under reduced pressure. The mixture was purified by column chromatography to obtain B-10-1 (160 mg, 78%).

[0302] B-10-1 was brominated with NBS and then ammonolysis was carried out to prepare B-10. For the specific method, refer to the method for producing B-1-A(B) from B-1-9. Example 1: Preparation of Compound 1

[0303] [ka]

[0304] 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 a flask, purged with nitrogen, heated to 95 °C and reacted for 2.5 hours, after which TLC showed that the reaction was complete. The reaction solution was diluted with ethyl acetate, directly added and mixed with silica gel, and then purified with a silica gel column to obtain 182 mg of product C-1-B, with a yield of 72%.

[0305] Compound C-1-B (182 mg, 0.260 mmol) and tetrahydrofuran (4 mL) were added to the flask, and TBAF (1 M, 0.39 mL) was added to the reaction solution, and the reaction solution was stirred at room temperature for 1.5 hours. TLC showed that the reaction was complete. The reaction solution was directly purified on a preparative silica gel plate (DCM / MeOH=15 / 1) to give 70 mg of product 1-B, with a yield of 58%.

[0306] The structure of the product was characterized by nuclear magnetic resonance and mass spectroscopy, with the results being as follows: 1 H NMR (400 MHz, d6-DMSO) δ 1.56-1.60 (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 = 11.8 Hz, 3.2 Hz), 4.11 (1H, dd, J = 11.8 Hz, 1.8 Hz), 4.59 (1H, t, J = 5.8 Hz), 6.03 (2H, brs), 7.07 (1H, d, J = 5.0 Hz), 7.20 (1H, ddd, J = 7.4 Hz, 4.9 Hz, 1.0 Hz), 7.63 (2H, dd, J = 10.4 Hz, 5.4 Hz), 7.85-7.90 (1H, m), 7.98-8.02 (2H, m), 8.21 (1H, d, J = 8.4 Hz), 8.41-8.43 (1H, m), 10.97 (1H, s). MS(ESI) m / z (M+H) + :463.0.

[0307] 1-A was prepared using A-1 and B-1-A according to the synthesis method for 1-B.

[0308] The structure of the product was characterized by nuclear magnetic resonance and mass spectroscopy, with the results being as follows: 1H NMR (400 MHz, d6-DMSO) δ 1.38-1.50 (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 = 11.0 Hz), 4.07-4.10 (1H, m ), 4.69 (1H, t, J = 5.5 Hz), 6.02 (2H, s), 7.07 (1H, d, J = 5.0 Hz), 7.20 (1H, dd, J = 6.9 Hz, 5.2 Hz ), 7.61 (1H, t, J = 7.9 Hz), 7.76 (1H, d, J = 5.0 Hz ), 7.83-7.91 (1H, m), 7.95-8.04 (2H, m), 8.21 (1H, d, J = 8.4 Hz ), 8.42 (1H, d, J = 3.8 Hz ),10.97 (1H, s). MS(ESI) m / z (M+H) + :463.1. Example 2: Preparation of Compound 2

[0309] [ka]

[0310] 2-A was prepared by reacting A-5 with B-1-A in the same manner as in the synthesis of 1-B.

[0311] The structure of the product was characterized by nuclear magnetic resonance and mass spectroscopy, with the results being as follows: 1H NMR (400 MHz, d6-DMSO) δ 1.41-1.51 (1H, m), 1.50-1.78 (1H, m), 1.85-1.97 (1H, m), 2.13-2.15 (1H, m), 3.35-3.49 (4H, m), 3.65 (1H, t, J = 11.0 Hz), 4.10 (1H, ddd, J = 11.0 Hz, 3.6 Hz, 1.6 Hz), 4.69 (1H, t, J = 5.6 Hz), 6.13 (2H, brs), 7.09 (1H, d, J = 4.9 Hz), 7.19 (1H, dd, J = 6.9 Hz, 5.2 Hz), 7.76 (3H, dd, J = 9.5 Hz, 6.7 Hz), 7.83-7.90 (1H, m), 8.16 (2H, d, J = 8.4 Hz), 8.23 ​​(1H, d, J = 8.4 Hz), 8.41 (1H, dd, J = 4.8 Hz, 1.0 Hz), 10.84 (1H, s). MS(ESI) m / z (M+H) + :445.2.

[0312] Compound 2-A was separated by SFC to give 2-A-P1 (early peak) and 2-A-P2 (late peak).

[0313] Preparative SFC conditions: Instrument: SFC-80 (Thar, Waters) Column: CHIRALCEL OJ (30×250mm 5μm) (Daicel) Column temperature: 35℃ Fluid phase: A=CO2 cosolvent B= ETOH Cycle time: 12.5 min Operating time: 21 min

[0314] [Table 13]

[0315] 2-B was prepared by reacting A-5 with B-1-B in the same manner as in the synthesis of 1-B.

[0316] The structure of the product was characterized by nuclear magnetic resonance and mass spectroscopy, with the results being as follows: 1 H NMR (400 MHz, d6-DMSO) δ 1.58-1.63 (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, 3.2 Hz), 4.10 (1H, d, J = 10.1 Hz), 4.60 (1H, t, J = 5.2 Hz), 6.14 (2H, brs), 7.09 (1H, d, J = 4.9 Hz), 7.18 (1H, dd, J = 6.9 Hz, 5.3 Hz), 7.63 (1H, d, J = 5.0 Hz), 7.76 (2H, d, J = 8.3 Hz), 7.84-7.88 (1H, m), 8.16 (2H, d, J = 8.3 Hz), 8.23 ​​(1H, d, J = 8.3 Hz), 8.41 (1H, dd, J = 4.8 Hz, 1.0 Hz), 10.84 (1H, s). MS(ESI) m / z (M+H) + :445.2. Example 3: Preparation of Compound 3

[0317] [ka]

[0318] Compound 2-B (50 mg, 0.113 mmol), NCS (16.5 mg, 0.124 mmol) and glacial acetic acid (1 mL) were added to a flask, and the reaction solution was heated to 80° C. for 2 hours. The reaction solution was concentrated and dried under reduced pressure, and aqueous sodium bicarbonate solution was added, and extracted twice with ethyl acetate. The organic phases were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, suction filtered, concentrated and dried under reduced pressure, and purified on a silica gel preparative plate to give 28 mg of product 3, with a yield of 52%.

[0319] MS(ESI) m / z (M+H) + :479.2. Examples 4 to 119: Preparation of Compounds 4 to 119

[0320] Compounds 4 to 119 were prepared using different intermediates in the method for producing compound 1-B or 3, and the numbers, structural formulas, MS and 1 H-NMR data is shown in Table 14.

[0321] [Table 14] JPEG0007660800000104.jpg247170JPEG0007660800000105.jpg255162JPEG00076608000001 06.jpg255167JPEG0007660800000107.jpg255162JPEG0007660800000108.jpg244170JPEG000 7660800000109.jpg244170JPEG0007660800000110.jpg255167JPEG0007660800000111.jpg2 55162JPEG0007660800000112.jpg255159JPEG0007660800000113.jpg255157JPEG0007660800 000114.jpg237170JPEG0007660800000115.jpg255167JPEG0007660800000116.jpg255167JP EG0007660800000117.jpg255161JPEG0007660800000118.jpg255163JPEG0007660800000119. jpg255166JPEG0007660800000120.jpg249170JPEG0007660800000121.jpg255157JPEG000766 0800000122.jpg255158JPEG0007660800000123.jpg255164JPEG0007660800000124.jpg94170

[0322] Example 120: Preparation of Compound 120

[0323] [ka]

[0324] 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 a flask. The reaction was stirred at room temperature overnight. It was poured into water and extracted twice with ethyl acetate. The combined organic phase was washed with saturated brine, concentrated and dried under reduced pressure, and then purified by silica gel column to give 1.90 g of product 120-1, with a yield of 59%.

[0325] Compound 120-1 (1.90 g, 4.95 mmol) and tetrahydrofuran (20 mL) were added to the flask, cooled in an ice bath, and lithium aluminum hydride (376 mg, 9.91 mmol) was added to the reaction solution in several portions. The reaction solution was warmed to room temperature and reacted for 3 hours. The reaction solution was cooled again in an ice bath, and water (380 mg), 15% NaOH aqueous solution (380 mg), and water (1.14 g) were slowly added dropwise in succession to quench the reaction. The resulting suspension was suction filtered and washed with DCM / MeOH (10 / 1), and the filtrate was concentrated and dried under reduced pressure, and then purified on a silica gel column to obtain 200 mg of product 120-2, with a yield of 31%.

[0326] 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 a flask. The reaction was stirred at room temperature for 4 hours. The reaction was poured into water and extracted with ethyl acetate three times, the combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to dryness under reduced pressure to give 464 mg of product 120-3, the yield was 100%. The product was not further purified.

[0327] A flask was charged with 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). The reaction was heated to 95°C and stirred for 4 h. The reaction was cooled, concentrated to dryness under reduced pressure and then purified on a silica gel column to give 540 mg of product 120-4, with a yield of 66%.

[0328] Compound 120-4 (388 mg, 0.76 mmol), DMF (4 mL), imidazole (78 mg, 1.14 mmol), DMAP (10 mg, 0.076 mmol) and t-butyldiphenylchlorosilane (210 mg, 0.76 mmol) were added to the flask. The reaction was heated to 60°C and stirred overnight. TLC showed that a lot of starting material remained. Additional imidazole (150 mg), DMAP (40 mg) and t-butyldiphenylchlorosilane (100 mg) were added to the reaction, which was then heated to 80°C and reacted for 2 hours. Additional t-butyldiphenylchlorosilane (200 mg) was added to the reaction, and after 2 hours, TLC showed that the reaction was complete. The reaction mixture was cooled, poured into water, and extracted with ethyl acetate three times. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and dried under reduced pressure, and then purified on a silica gel column to give 650 mg of product 120-5, with a yield of 100%.

[0329] 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 a flask. The reaction was stirred at room temperature for 4 h. The reaction was slowly poured into aqueous NaHCO3 solution, extracted twice with ethyl acetate, and the combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure to give 612 mg of product 120-6, 98% yield. The product was not further purified.

[0330] A flask was charged with compound 120-6 (612 mg, 0.85 mmol), acetonitrile (6 mL) and N,N'-carbonyldiimidazole (280 mg, 1.71 mmol). The reaction was heated to 80 °C and stirred overnight. The reaction was cooled, concentrated to dryness under reduced pressure and then purified on a silica gel column to give 470 mg of product 120-7, with a yield of 74%.

[0331] A flask was charged with compound 120-7 (470 mg, 0.63 mmol), dichloromethane (15 mL), 4-phenoxyphenylboronic acid (271 mg, 1.27 mmol), copper acetate (115 mg, 0.63 mmol), 4A molecular sieves (500 mg) and triethylamine (192 mg, 1.90 mmol). The reaction was stirred at room temperature for 36 h. The reaction was filtered through diatomaceous earth and washed with ethyl acetate, and the filtrate was concentrated and dried under reduced pressure, then purified on a silica gel column to give 240 mg of product 120-8, with a yield of 41%.

[0332] Compound 120-8 (260 mg, 0.29 mmol), dichloromethane (4 mL) and trifluoroacetic acid (4 mL) were added to the flask. The reaction was heated to 50° C. and stirred for 3 h. The reaction was cooled, concentrated to dryness under reduced pressure, added with aqueous NaHCO3, extracted twice with ethyl acetate, the combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was dissolved in tetrahydrofuran (2 mL), added with TBAF (1M, 0.2 mL) and stirred at room temperature for 1 h. The reaction was directly purified on a silica gel prep plate to give 40 mg of product 120, with a yield of 30%.

[0333] The structure of the product was characterized by nuclear magnetic resonance and mass spectroscopy, with the results being as follows: 1H NMR (400 MHz, d6-DMSO) δ 1.37-1.53 ​​(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 = 5.7 Hz ), 4.76-4.83 (2H, m), 6.94 (0.5H, d, J = 5.6 Hz), 7.13 (4H, t, J = 8.5 Hz), 7.18-7.25 (1.5H, m), 7.40-7.48 (4H, m), 7.74 (1H, t, J = 5.6 Hz). MS(ESI) m / z (M+H) + :433.2.

[0334] Examples 121-138: Preparation of Compounds 121-138 Compounds 121 to 138 were prepared using different intermediates in the method for producing compound 1-B or 3, and the numbers, structural formulas, MS and 1 H-NMR data is shown in Table 15.

[0335] [Table 15] JPEG0007660800000127.jpg255170JPEG0007660800000128.jpg226170JPEG0007660800000129.jpg240170 JPEG0007660800000130.jpg255167JPEG0007660800000131.jpg242170JPEG0007660800000132.jpg255163

[0336] Drug efficacy test Test Example 1: Testing the activity of BTK-inhibited kinases in vitro 1. Compound Preparation The compound powder was dissolved in 100% DMSO to prepare a 10 mM stock solution, which was then frozen and stored at -20°C in the dark.

[0337] 2. Kinase reaction process (1) 1× kinase buffer was prepared.

[0338] (2) Preparation of compound concentration gradient: The test compound was diluted to a test concentration of 1 μM in 100% DMSO solution at 100 times the final concentration in the 384 source plate, and the compound was diluted 3 times to 10 concentrations. 250 nL of the compound at 100 times the final concentration was transferred to the destination plate OptiPlate-384F using a liquid separator Echo550.

[0339] (3) A kinase solution was prepared at 2.5 times the final concentration using 1× kinase buffer.

[0340] (4) 10 μL of a kinase solution at 2.5 times the final concentration was added to each of the compound wells and the positive control wells, and 10 μL of 1× kinase buffer was added to the negative control wells.

[0341] (5) After centrifugation at 1,000 rpm for 30 seconds, the reaction plate was shaken to mix uniformly and then incubated at room temperature for 10 minutes.

[0342] (6) A mixed solution of ATP and kinase substrate 2 was prepared at a final concentration of 5 / 3 in 1× kinase buffer.

[0343] (7) 15 μL of a mixed solution of ATP and substrate at 5 / 3 times the final concentration was added to initiate the reaction.

[0344] (8) The 384-well plate was centrifuged at 1,000 rpm for 30 seconds, shaken to mix uniformly, and then incubated at room temperature for 10 minutes.

[0345] (9) The kinase reaction was stopped by adding 30 μL of stop detection solution, centrifuged at 1000 rpm for 30 seconds, and shaken to mix uniformly.

[0346] (10)The conversion rate was read using the Caliper EZ Reader.

[0347] 3. Data Analysis Calculation formula:

[0348]

number

[0349] Where: %_sample is the conversion reading of the sample, %_min is the average of the negative control wells and represents the conversion reading of the wells without enzyme activity, and %_max is the average of the positive control wells and represents the conversion reading of the wells without compound inhibition.

[0350] Dose-response curve fitting The log value of concentration is plotted on the X-axis and the percentage inhibition rate on the Y-axis. A dose-response curve was fitted using the analysis software GraphPad Prism 5 [log(inhibitor) vs. response - variable slope] to calculate the IC50 value of each compound against the enzyme activity using the following formula.

[0351] Calculation formula: Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)×HillSlope)) Here, Y: inhibition rate (%), X: concentration, Top: maximum inhibition rate, Bottom: minimum inhibition rate, HillSlope: slope.

[0352] The inhibitory activity of the compounds of the present invention against BTK wild-type and BTK mutant C481S kinase is shown in Table 16. IC50:A≦5 nM; <B≦20 nM;20 nM<C≦100 nM;100 nM<D≦1000 nM;E> 1000 nM.

[0353] [Table 16] JPEG0007660800000135.jpg166170

[0354] Test Example 2: Experiment on metabolic stability of liver microsomes

[0355] 1. 10μL of test sample or control working solution and 80μL of microsome working solution (concentration of liver microsomal protein was 0.5mg / mL) were added to the wells of T0, T5, T10, T20, T30, T60 and NCF60 samples, and only microsome working solution was added to the blank 60 well. Then, the blank 60 samples, T5, T10, T20, T30 and T60 except for T0 and NCF60 were placed in a 37℃ water bath and pre-incubated for about 10 minutes.

[0356] 2. To the T0 sample, 300 μL of a stop solution (acetonitrile solution containing 200 ng / mL tolbutamide and 200 ng / mL labetalol) was first added, and then 10 μL of the NADPH regenerating system working solution was added.

[0357] 3. After pre-incubating Blank 60, T5, T10, T20, T30 and T60 in the incubation plate, 10 μL of NADPH regenerating system working solution was added to each sample well to start the reaction, and 10 μL of 100 mM potassium phosphate buffer was added to the NCF60 sample well.

[0358] 4. After incubation for an appropriate period of time (e.g., 5, 10, 20, 30 and 60 minutes), the reaction was stopped by adding 300 μL of stop solution to each test product sample well and control product sample well on the Blank 60, T5, T10, T20, T30, T60 and NCF60 plates.

[0359] 5. All sample plates were shaken well and centrifuged at 4000 rpm for 20 minutes, and 100 μL of the supernatant of each test sample or control sample was diluted with 300 μL of pure water and subjected to LC-MS / MS analysis.

[0360] 6. Data Analysis Based on First-order Elimination Kinetics 1 / 2and C.L. int(mic) The kinetics of first-order elimination was calculated by the following equation:

[0361]

number

[0362] The results of human and rat hepatic microsomal metabolic stability studies are shown in Table 17.

[0363] [Table 17]

[0364] Test Example 3: Pharmacokinetics test

[0365] For the pharmacokinetic study, each test compound was administered orally to SD rats once (10 mg / kg, 3 rats per group). The test compound was dissolved in 5% DMSO + 10% solutol + 85% saline, vortexed for 1-2 minutes, and sonicated for 5-10 minutes to prepare a colorless, transparent, clear administration solution. The animals were fasted overnight before oral administration and resumed eating 4 hours after administration. After oral administration to SD rats, pharmacokinetic samples were collected by orbital blood sampling. However, the collection times were 0.25 h, 0.5 h, 1 h, 2 h, 2.5 h, 3 h, 4 h, 6 h, 8 h, and 10 h after administration, and three whole blood samples were collected at each time point with a collection volume of approximately 0.2-0.3 mL. After blood samples were collected, they were placed on ice immediately and the plasma was centrifuged within 15 minutes (centrifugation conditions: 8000 rpm, 1 min, room temperature). The collected plasma was stored at −20°C until analysis. 20 μL of plasma samples were placed into a 1.6 mL 96-well deep-well plate, 200 μL of working internal standard solution was added (same volume of solvent was added instead of internal standard to the blank), vortex mixed for 1 min, centrifuged at 5800 rpm for 10 min, and 100 μL of the supernatant was added to a 96-well injection plate and subjected to LC-MS / MS for analysis.

[0366] The results of pharmacokinetic studies of some compounds of the invention are shown in Table 18 below.

[0367] [Table 18]

[0368] Test Example 4: Test for inhibitory activity against cell proliferation in vitro

[0369] 1.Cell culture The cells were cultured in 1640 medium supplemented with 10% inactivated FBS and 1% double antibody, and cultured at 37°C under 5% CO2 conditions.

[0370] 2. Cell Plating (1) The cells were cultured in the usual manner until the cell saturation reached 80% to 90%, and when the required number of cells was reached, the cells were harvested.

[0371] (2) Resuspend in the corresponding medium, count, and prepare a cell suspension of appropriate density.

[0372] (3) The cell suspension was added to a 96-well plate at 100 μL per well.

[0373] (4) The cells were cultured overnight in a 37°C, 5% CO2 incubator.

[0374] 3. Compound Preparation (1) Each test compound was diluted with DMSO to prepare a mother solution with a final concentration of 20 mM.

[0375] (2) The mother liquor was diluted 10-fold with DMSO from 20 mM to 2 mM, and then diluted 3-fold from 2 mM to give nine concentrations.

[0376] (3) Blank control wells contained cells and 0.5% DMSO and served as high readout control wells.

[0377] (4) Wells containing only medium without cells served as low reading control wells.

[0378] 4. Treatment of Cells with Compounds (1) 24 hours after plating the cells, the compounds were applied alone, supplemented with 99 μL of growth medium per well, then added 1 μL of the compound prepared in step 3, gently shaken to ensure uniform mixing, and then placed in a 37°C, 5% CO2 incubator.

[0379] (2) The cell plate was left in an incubator for 72 hours.

[0380] 5. Detection by CTG method (1) The cell measurement plate was left to stand at room temperature for 30 minutes, and 100 μL of medium per well was discarded.

[0381] (2) 100 μL of CTG reagent (CelltiterGlo kit) was added to each well, shaken for 2 minutes on a high-speed shaker, and then left at room temperature in the dark for 30 minutes.

[0382] (3) The chemiluminescence signal values ​​were read using an Envision instrument.

[0383] 6. Data Analysis IC with GraphPad Prism 8 software 50 Calculate the IC of the compound using the following nonlinear fitting equation: 50 (half maximal inhibitory concentration) was obtained and the results are shown in the table below.

[0384] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)×HillSlope) X: log value of compound concentration, Y: inhibition rate (%inhibition) % inhibition = (high readout control well reading - compound well reading) / (high readout control well reading - low readout control reading) x 100

[0385] [Table 19] [Table 20] [Table 21] [Table 22]

[0386] Test Example 5: HER2 kinase activity test

[0387] 1. Her2 kinase measurement procedure 1) Prepare 1x kinase reaction buffer: 1 volume of 5x kinase reaction buffer plus 4 volumes of water, 1 mM dithiothreitol, 5 mM magnesium chloride, 1 mM manganese chloride, 12.5 mM SEB.

[0388] 2) 100 nl of the diluted compound working solution was transferred to each well of a reaction plate (784075, Greiner) in an Echo 550. The reaction plate was sealed with sealing film and centrifuged at 1000 g for 1 minute.

[0389] 3) A 1 ng / μL Her2 kinase solution was prepared using 1× kinase reaction buffer.

[0390] 4) 5 μL of the kinase solution prepared above was added to each well of the reaction plate. The plate was sealed with a sealing film, centrifuged at 1000 g for 1 minute, and left at room temperature for 10 minutes.

[0391] 5) A mixture of 2x kinase substrate and ATP was prepared in 1x kinase reaction buffer, where the 2x Her2 kinase substrate was 2μM TK-substrate-biotin and 4μM ATP.

[0392] 6) 5 μL of a mixture of 2×TK-substrate-biotin and ATP was added to the reaction plate, and the plate was centrifuged at 1000 g for 30 seconds to initiate the reaction.

[0393] 7) For Her2 kinase measurement, the reaction was carried out at room temperature for 50 minutes.

[0394] 8) A mixture of Sa-XL665 (125 nM) and TK-antibody-cryptate was prepared in HTRF detection buffer.

[0395] 9) 10 μL of a mixture of Sa-XL665 and TK-antibody-cryptate was added to each well, centrifuged at 1000 g for 30 seconds, and reacted at room temperature for 1 hour.

[0396] 10) Fluorescence signals were read at 615 nm (cryptate) and 665 nm (XL665) using an Envision 2104.

[0397] 2. Data Analysis 1) The inhibition rate was calculated using the following formula.

[0398]

number

[0399] 2) IC50 calculation and compound dose-response curve fitting: The IC50 of the compound was obtained using the following nonlinear fitting equation in GraphPad 6.0:

[0400] Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)×HillSlope)) X: log value of compound concentration; Y: compound inhibition rate (%)

[0401] [Table 23]

[0402] Test Example 6: Blood-brain barrier permeability test

[0403] For each test compound, a single oral administration pharmacokinetic study was conducted using SD rats, with a dose of 10 mg / kg, and 9 animals per group. The test compound was dissolved in 5% DMSO + 10% solutol + 85% saline, vortexed for 1-2 minutes, and sonicated for 5-10 minutes to prepare a colorless, transparent, clear administration solution. The animals were fasted overnight before administration, and approximately 0.2-0.3 mL of blood was collected from the orbit of three SD rats in each group 1 h, 2 h, and 4 h after administration. The blood samples were placed on ice immediately after collection, and the plasma was centrifuged within 15 minutes (centrifugation conditions: 8000 rpm, 1 min, room temperature). The collected plasma was stored at -20°C until analysis. Cerebrospinal fluid and brain tissue were collected immediately after blood collection. Cerebrospinal fluid was extracted by dural puncture with a microinjector under direct vision, i.e., after anesthesia with chloral hydrate, the head was fixed, the hair was cut at the back, a transverse incision (2 cm) was made at the line connecting both ears, the muscle layer of the neck and base of the skull was bluntly cut to expose the foramen magnum, and approximately 100 μl of cerebrospinal fluid was collected with a 100 μl microinjector and stored at -20°C until analysis. Immediately afterwards, the rat was sacrificed, the head was cut, and the brain tissue was extracted, the capillaries on the surface were stripped, the weight was measured, 3 volumes of cold saline were added, homogenized with a homogenizer for 1 minute, and stored at -20°C until analysis. 200 μL of working internal standard solution was added to 20 μL of plasma sample and brain homogenized sample (except blank, where the same volume of solvent was added instead of internal standard), vortex mixed for 1 min, centrifuged at 13500 rpm for 10 min, 100 μL of supernatant was collected and analyzed by LC-MS / MS. 60 μL of working internal standard solution was added to 20 μL of cerebrospinal fluid sample (except blank, where the same volume of solvent was added instead of internal standard), vortex mixed for 1 min, centrifuged at 13500 rpm for 10 min, 50 μL of supernatant was collected and analyzed by LC-MS / MS.

[0404] [Table 24]

[0405] Test Example 7: Testing the TMD8 drug efficacy model

[0406] Human diffuse large B lymphoma TMD8 cells were cultured in a monolayer in vitro. The culture conditions were RPMI1640 medium supplemented with 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin, and the cells were cultured at 37°C in a 5% CO2 incubator. Subculture was performed twice a week by regular digestion with trypsin-EDTA. When the cell saturation reached 80%-90% and the required number was reached, the cells were harvested, counted, and inoculated. 0.2 ml (1x10 7 1) TMD8 cells (containing Matrigel, volume ratio 1:1) were inoculated subcutaneously into the right hind back of each mouse, and the average tumor volume was approximately 137 mm 3 When the tumor diameter reached 100 mm, the mice were divided into groups and administered the drug. The tumor diameter was measured twice a week with a caliper. The tumor volume was calculated as V = 0.5a × b 2 where a and b represent the long and short diameters of the tumor, respectively.

[0407] The results are shown in Figures 1 and 2. From the efficacy model of TMD8 mice with subcutaneously implanted tumors shown in Figure 1, it was found that under the same 10 mg / kg dose condition, the two compounds of Example 118 and Example 89-P1 had significantly better tumor inhibitory effects than the drug ARQ-531 in Phase II clinical trials and the commercially available drug ibrutinib. From the efficacy model of subcutaneously implanted tumors in TMD8 mice shown in Figure 2, it was found that under the same 20 mg / kg dose condition, the compounds of Example 111-P1 and Example 125 had significantly better tumor inhibitory effects than tirabrutinib. In particular, in Example 111-P1, in terms of tumor inhibition rate, the TGI of Example 111-P1 is 93%, which is about twice that of tirabrutinib, and it almost completely controls tumor growth and has a very significant drug efficacy advantage.

[0408] Test Example 8: Test of DOHH-2-Luc intracranial tumor efficacy model 1.Cell culture DOHH-2-luc tumor cells were cultured in vitro in RPMI1640 medium containing 10% fetal bovine serum and 500 ng / mL puromycin in an incubator at 37°C and 5% CO2. The medium was replenished or replaced every 2-3 days, and the number of passages was not more than 4-5. Tumor cells in the logarithmic growth phase were used for in vivo tumor inoculation.

[0409] 2. Tumor Cell Inoculation and Grouping After anesthetizing the animal with an intramuscular injection of Zoletil, the animal was fixed in a supine position on an operating table, and the skin on the top of the head was disinfected with iodine and 75% alcohol, respectively. Approximately 0.5 cm of the skin was incised along the midline of the head to expose the coronal and sagittal lines. Using a brain stereotaxic apparatus, a position was identified at a distance of approximately 0.5 to 1.0 mm above the coronal line and approximately 2 mm to the right of the sagittal line. A hole was made with a 1 mL injection needle, and a microinjector was inserted vertically at that position to a depth of 3 mm, and 3 × 10 DOHH-2-luc tumor cells were injected. 5 2 μL / mL suspension was slowly injected (approximately 1 min) and the needle was held for 1 min. After the needle was removed, the needle hole was promptly sealed with bone wax and the wound was sutured with a skin stapler. Approximately 7 days after tumor inoculation, the animals were randomly divided into 5 groups of 5 animals each based on the animal weight and the optical signal intensity at the tumor site.

[0410] 3. Image Analysis Using a small animal in vivo imaging system IVIS Lumina III (Perkin Elmer), the mice were imaged once or twice a week depending on their condition, and the signal intensity of bioluminescence imaging (BLI, unit: photons / s) at the inoculation site of mouse tumor cells was monitored as the main indicator for evaluating tumor growth and drug efficacy. The specific procedures were as follows:

[0411] After intraperitoneal injection of D-luciferin (15 mg / mL, 5 μL / g of experimental animal body weight) into mice, the animals were anesthetized by inhalation of 1% to 2% isoflurane, and D-luciferin was injected for 10 min, after which the animals were imaged by IVIS Lumina III. Data were analyzed by the Living Image software (Perkin Elmer) to calculate the optical signal intensity within the ROI (regions of interest) of each animal.

[0412] The results are shown in Figures 3 and 4. Figure 3 shows that in the study of mouse intracerebral DOHH2 tumor model, under the same dose condition of 30mg / kg (BID), the compounds of Example 111-P1 and Example 125 have significantly better tumor inhibitory effect than tirabrutinib, have very significant pharmacological advantage, and no side effects were observed after 21 days of administration.

[0413] Figure 4 is the fluorescence image of all the animals after imaging, where the size of the tumor in the brain is represented by color and area size, and the redder the color, the larger the tumor.These images show that at the same dose, the compounds of Example 111-P1 and Example 125 have very good tumor inhibition effect, and there are few red areas, indicating that the animals in these two groups have small tumors in the brain, whereas all the animals in the model group and tirabrutinib group have large red areas, indicating that the tumors are large.

[0414] From the above examples, it can be seen that the compounds of the present invention as BTK protein kinase inhibitors having the structure of formula I, preferably the structure of formula II, have strong inhibitory effects on wild-type BTK and mutant BTK (C481S), have good pharmacokinetic properties, and can be used to manufacture medicines for treating diseases caused by overexpression of BTK kinase. Some of these compounds are significantly superior to the listed BTK inhibitors ibrutinib, tirabrutinib, and ARQ-531 in phase II clinical trials in TMD8 subcutaneous tumor efficacy model experiments.

[0415] Some compounds of the present invention are significantly superior to commercially available drugs Tirabrutinib and Tucatinib in terms of blood-brain barrier permeability, liver microsome metabolic stability, pharmacokinetics, etc. The efficacy of some compounds is very good, as demonstrated by the data of DOHH-2-Luc brain efficacy model and blood-brain barrier permeability. The compounds of the present invention can be used to manufacture medicines for treating diseases caused by overexpression of BTK or HER2 kinase, particularly brain diseases.

[0416] The compounds according to the present invention or their stereoisomers, solvates, hydrates, pharma- ceutically acceptable salts or cocrystals can be used for the manufacture of medicaments for treating any one or more diseases selected from autoimmune diseases, inflammatory diseases, thromboembolic diseases, allergies, infectious diseases, proliferative diseases and cancer, and it is hoped to provide new and improved treatment regimens.

[0417] It should be noted that the above are merely preferred embodiments of the present invention, and those skilled in the art can make various modifications to these embodiments without departing from the technical principles of the present invention, and these modifications should also be considered as within the scope of the present invention to be protected.

Claims

1. A compound as a BTK inhibitor, characterized in having a structure represented by Formula III or Formula IV, or in the form of a tautomer, meso isomer, racemate, enantiomer, diastereoisomer, or mixture thereof, a pharma- ceutically acceptable hydrate, solvate, or salt thereof. 【Chemistry 1】 (Wherein, X is 【Chemistry 2】 wherein R 9 , R 13 are independently selected from hydrogen, a hydroxyl group, an amino group, a cyano group, and a C1-C3 alkyl group; R 1 is selected from hydrogen, an amino group, and a C1-C6 alkyl group; R2 is selected from hydrogen and halogen; R 3 and R 4 are selected from hydrogen, a C1-C6 alkyl group; R 6 is selected from hydrogen, halogen, and a C1-C6 alkyl group; R 8 are independently selected from hydrogen, fluorine, chlorine, bromine, a cyano group, an amino group, a C1-C3 alkyl group, a C1-C3 alkyloxy group, and a C3-C6 cycloalkyl group; m is selected from the integers 0, 1 and 2, with the proviso that when m is 0, there is no covalent bond between two adjacent ring carbon atoms of --(CH 2 ) m --; n is selected from the integers 0 and 1; n1 is selected from 0 and 1; n2 is selected from 0, 1, 2, and 3. (However, within the above range (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; (If any of the above is included, exclude the items that are included)

2. A compound characterized in that the structure of the compound is one selected from the following: 【Chemistry 3】 【Chemistry 4】 【Chemistry 5】 【Chemistry 6】 【Chemistry 7】 【Chemistry 8】 【Chemistry 9】 【Chemistry 10】 【Chemistry 11】 【Chemistry 12】 【Chemistry 13】 【Chemistry 14】 【Chemistry 15】 【Chemistry 16】 【Chemistry 17】

3. A pharmaceutical composition comprising, as an active ingredient, one or a combination of a plurality of compounds selected from the compound according to claim 1 or 2, or a stereoisomer, solvate, hydrate, or pharma- ceutically acceptable salt thereof.

4. 3. Use of a compound according to claim 1 or 2, or a stereoisomer, solvate, hydrate, or pharma- ceutically acceptable salt thereof, in the manufacture of a protein kinase inhibitor, further comprising said kinase inhibitor being a BTK inhibitor.

5. 3. Use of the compound of claim 1 or 2, or a stereoisomer, solvate, hydrate, or pharma- ceutically acceptable salt thereof, in the manufacture of a medicament for treating a disease caused by overexpression of BTK kinase.

6. Use of a compound according to claim 1 or 2, or a stereoisomer, solvate, hydrate, or pharma- ceutically acceptable salt thereof, in the manufacture of a medicament for treating any one or more diseases selected from an autoimmune disease, an inflammatory disease, a thromboembolic disease, an allergy, an infectious disease, a proliferative disease, and cancer.

7. The disease is selected from the group consisting of arthritis, rheumatoid arthritis, urticaria, vitiligo, organ transplant rejection, ulcerative colitis, Crohn's disease, dermatitis, asthma, Sjogren's syndrome, systemic lupus erythematosus, multiple sclerosis, idiopathic platelet-reduced purpura, skin rash, antineutrophil cytoplasmic antibody-associated vasculitis, pemphigus, 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, renal cancer, and pancreatic cancer.

7. The use according to claim 6, characterized in that the cancer is selected from the group consisting of lymphoma, hairy cell carcinoma, nasopharyngeal carcinoma, pharyngeal carcinoma, colon 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 carcinoma, lung adenocarcinoma, bone cancer, colon cancer, adenoma, pancreatic cancer, adenoma, thyroid cancer, follicular carcinoma, Hodgkin's leukemia, bronchial carcinoma, thyroid cancer, uterine cancer, cervical cancer, multiple myeloma, acute myeloid leukemia, chronic myeloid leukemia, lymphocytic leukemia, chronic lymphocytic leukemia, osteomyeloma, non-Hodgkin's lymphoma, and primary macroglobulinemia.

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