Multi-target anti-tumor compound and its preparation method and application

Abstract

A new class of small molecule compounds with multiple kinase inhibitory activity has a structure shown in general formula II. The compounds shown in general formula II have good efficacy and safety in preventing or treating various cancer indications such as lung cancer and lymphoma.

Description

[0001] This application claims the priority of the Chinese patent application filed with the State Intellectual Property Office of China on April 2, 2020, with application number 202010252669.5 and invention name “Multi-target anti-tumor compounds and preparation methods and applications thereof”, the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The present invention belongs to the field of pharmaceutical chemistry, and specifically relates to a new class of small molecule compounds with multiple kinase inhibitory activity and a preparation method thereof, as well as the use of such compounds in preventing and treating tumors. Background Art

[0003] Gene fusion is a driving factor for many malignant tumors. Structurally, they fuse with upstream genes and maintain a complete tyrosine kinase domain, independent of ligand dimerization, resulting in continuous activation of downstream signals to promote tumor cell growth and proliferation. With the continuous maturity and improvement of technologies such as second-generation sequencing and RT-PCR, many clinically relevant fusion genes have been detected. Including NTRK1, NTRK2, NTRK3 (encoding TRKA, TRKB and TRKC proteins), ALK, ROS1, etc.

[0004] Tropomyosin receptor kinase (TRK) is a class of nerve growth factor receptors, including three receptors, TRKA, TRKB and TRKC, encoded by NTRK1, NTRK2 and NTRK3 genes. They are widely expressed in neuronal tissues and are involved in the maintenance, signal transduction and survival of neuronal cells. TRK is a transmembrane receptor composed of an extracellular ligand binding domain, a transmembrane domain and an intracellular ATP binding domain. The extracellular domains of TRKA, TRKB and TRKC show a high degree of structural similarity, including two immunoglobulin-like motifs (Ig1 and Ig2), which are considered to contain the ligand binding site, and three leucine-rich 24-residue motifs (LRR1-3 motifs). The LRR1-3 motifs are flanked by two cysteine ​​clusters (C1 and C2) that are specific to TRK proteins and are not found in other subfamilies of receptor tyrosine kinases.

[0005] The TRKA is a high affinity receptor for nerve growth factor (hereinafter referred to as NGF), the TRKB is a high affinity receptor for brain-derived neurotrophic factor (BDNF) and neurotrophic factor (hereinafter referred to as NT)-4 / 5, and the TRKC is a high affinity receptor for NT-3. It is known that the activation of TRKA in peripheral nerves by NGF induces hyperalgesia, including nociceptive pain in osteoarthritis, chronic low back pain, rheumatoid arthritis, fractures, interstitial cystitis and chronic pancreatitis, neuropathic pain, and cancer pain accompanied by two of the above types of pain. At the same time, the literature shows that the activation of the BDNF / TrkB pathway is also involved in various types of pain, including inflammatory pain, neuropathic pain and surgical pain, so inhibitors of TRKA kinase and other TRK kinases can provide effective treatment for chronic pain states.

[0006] In addition, changes in the TRK signaling pathway, including gene fusion, protein overexpression or single nucleotide changes, have been found to be associated with many cancers. The cancers include neuroblastoma, ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, multiple myeloma, astrocytoma and medulloblastoma, glioma, melanoma, thyroid cancer, lung adenocarcinoma, large cell neuroendocrine tumors and colorectal cancer. Among them, the fusion of the NTRK gene is currently the most clear cause of cancer. TRK fusion protein can drive the spread and growth of TRK fusion tumors by activating PI3K, RAS / MAPK / ERK and PLC-γ pathways and triggering permanent signal cascades, independent of ligand dimerization and phosphorylation. The NTRK gene fusion can occur anywhere in the body, so TRK fusion cancers may occur in a variety of adult and pediatric solid tumors, including mammary analog secretory carcinoma (MASC), colon cancer, lung cancer, pancreatic cancer, thyroid cancer, and various sarcomas. NTRK gene fusions occur in approximately 1% to 3% of lung cancers, 0.5% to 1% of colon, breast, and thyroid cancers, and melanomas, and in more than 90% of salivary gland cancers, a type of juvenile breast secretory carcinoma, and infantile fibrosarcoma. Therefore, inhibitors of the TRK kinase family have utility in the treatment of cancer.

[0007] Anaplastic lymphoma kinase (ALK) is a 200kd receptor tyrosine kinase encoded by the ALK gene on chromosome 2p23. ALK belongs to the insulin receptor superfamily. ALK plays an important role in fetal development, but is not expressed in adult tissues other than brain tissue. ALK is considered to be an oncogene, and studies have found that it is abnormally expressed in B-cell lymphoma, inflammatory myofibroblastic tumor, some non-small cell lung cancer, renal cancer, colorectal cancer, and neuroblastoma. ALK gene rearrangement allows it to escape the control of the silent promoter, accompanied by transcription and translation activation protein of the fusion gene, and ALK fusion protein is mainly expressed in the cytoplasm, transducing mitosis, cell survival and anti-apoptosis signals to the nucleus through intracellular pathways to exert its effects. For example, in anaplastic large cell lymphoma (ALCL), chromosomal translocation causes NPM-ALK gene fusion. When fused with ALK, the dimerization domain of NPM causes constitutive dimerization and activation of ALK. At the same time, ALK recruits several adaptor proteins and stimulates multiple signaling pathways known to mediate tumor cell growth and survival, including STAT3, PLC-γ, RAS-ERK1,2, and PI3K-AKT, exerting its oncogenic activity. ALK translocation is also detected in about 5% of non-small cell lung cancer (NSCLC). Similar to the translocation of ALK in ALCL, the fusion protein in NSCLC shows constitutive ALK activity and promotes tumor growth and survival. In addition to chromosomal rearrangements, activating point mutations and amplifications have been reported in subsets of sporadic and familial neuroblastomas, which further expands the spectrum of tumors that depend on ALK activity. Tumors with abnormal ALK expression respond well to targeted small molecule inhibitors, such as crizotinib, which has been approved for clinical treatment of advanced and metastatic ALK-positive non-small cell lung cancer, and has good effects in inflammatory myofibroblastic tumor and anaplastic large cell lymphoma. Therefore, small molecule inhibitors of ALK are beneficial for therapeutic intervention in ALCL, NSCLC, neuroblastoma and other cancers that rely on ALK for growth and survival.

[0008] c-ROS proto-oncogene 1 (hereinafter referred to as ROS1) was originally discovered as a proto-oncogene and transforming sequence of the UR2 avian sarcoma virus. The ROS1 gene is located on chromosome 6q22.1, with a total of 46 exons and a total length of 127kb. The protein encoded by this gene is a type I integral membrane protein with tyrosine kinase activity and belongs to one of the members of the receptor tyrosine kinase family (RTK). The protein contains 2347 amino acids and acts as a growth or differentiation factor receptor. It consists of three domains: an extracellular domain containing an N-glycosylation site, a transmembrane domain encoded by exon 34, and a tyrosine kinase domain encoded by exons 36-41. ROS1 protein is most highly expressed in the kidney, followed by the cerebellum, peripheral nerve tissue, stomach, small intestine and colon, and is less expressed in other tissues.

[0009] The latest studies have shown that ROS1 is a new tumor-driving mutation gene, and its fusion is defined as a new molecular subtype in non-small cell lung cancer. The rearrangement of the ROS1 gene was first identified in human glioma cell lines, and was subsequently found in several other malignant tumors, such as cholangiocarcinoma, ovarian cancer, gastric cancer, and non-small cell lung cancer, among which the mutation frequency in non-small cell lung cancer is 1%-2%. When ROS1 fuses with other genes, the extracellular region is lost, the transmembrane region and the intracellular tyrosine kinase region are retained, and the rearrangement site mainly occurs in exons 32 to 36 of the ROS1 gene. ROS1 gene rearrangement produces a fusion protein with a constitutively active kinase domain, which activates downstream signaling pathways in cells that lead to oncogenic properties, including uncontrolled proliferation and resistance to cell death by prolonging tumor cell survival. These pathways include the Ras-ERK pathway for cell proliferation, as well as the JAK-STAT pathway and the PI3K / AKT pathway, which regulate cell survival (anti-apoptosis) and proliferation. ROS1 fusion protein can also activate the mTOR pathway, which is critical for regulating protein translation. Cancers with these pathways activated tend to be more aggressive, and invasion and metastasis lead to poor survival rates for patients. The ROS1 gene can fuse with multiple genes, the most important fusion partner of which is CD74. For example, in NSCLC, the ROS1 gene mainly fuses with SLC34A2 and CD74 and continuously activates the ROS1 tyrosine kinase region and downstream signaling pathways, thereby causing tumorigenesis.

[0010] RTK inhibitors have the potential to kill cancer cells that rely on dysregulated RTK activity without damaging normal tissues. Among them, TRK family protein tyrosine kinase inhibitors can be used to treat pain, inflammation, cancer and certain infectious diseases. Therefore, small molecule inhibitors targeting TRK, ALK, and ROS1 have promising therapeutic prospects. Summary of the invention

[0011] The present invention relates to a class of small molecule inhibitors targeting TRK, ALK, and ROS1, and specifically to a class of compounds shown in Formula II or their tautomers, mesomers, racemates, enantiomers, diastereomers, and mixtures thereof, and pharmaceutically acceptable salts, polymorphs, solvates, prodrugs, metabolites, isotope derivatives thereof, and pharmaceutical compositions comprising the compounds, which can be used to prevent or treat various types of cancers.

[0012]

[0013] Among them, Y 1 When it is carbon, Y 3 Nitrogen, Y 2 For carbon, Y 4 For carbon, Y 5 for carbon;

[0014] Y 1 When it is carbon, Y 2 For carbon, Y 3 Nitrogen, Y 4 For carbon, Y 5 for nitrogen;

[0015] Y 1 When Y is nitrogen, 4 Nitrogen, Y 2 For carbon, Y 3 For carbon, Y 5 for carbon;

[0016] Y 1 When Y is nitrogen, 2 Nitrogen, Y 3 For carbon, Y 4 For carbon, Y 5 for carbon;

[0017] R 0 for

[0018] X is carbon or nitrogen;

[0019] R 1 for -CF2-R 6 ,

[0020] R 2 For-NR a R b 、-SO2-NR a R b 、-S(O)-NR a R b 、-S(O)-(CH2)2-NR a Rb , 4-10 membered heterocyclic group, -C 4-10 Cycloalkyl, wherein the 4-10 membered heterocyclic group or the -C 4-10 Cycloalkyl includes compounds with monocyclic, bicyclic, tricyclic, bridged or spirocyclic structures;

[0021] The 4-10 membered heterocyclic group or -C 4-10 Cycloalkyl groups may be mono- or poly-substituted by: -OH, -C 1-3 Alkyl, -C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, 5-10 membered heteroaryl, -OC 1-4 Alkyl, -NR c R d 、-NH2、halogen、-C 1-3 haloalkyl, phenyl or cyano;

[0022] R 2 Preferably

[0023] R a and R b are independently hydrogen, -C 1-3 Alkyl, -C 3-6 Cycloalkyl, -C 1-4 Alkylene -OH, -C 1-3 Alkylene-C 3-9 Heterocycloalkyl, -C 1-3 Alkylene-C 5-6 Oxoheterocycloalkyl, -C 1-3 Alkylene-C 4-6 Cycloalkyl, -C(O)-(CH2) n -T, where the -C 3-9 Heterocycloalkyl includes monocyclic, bicyclic, tricyclic, bridged or spirocyclic compound groups, wherein the -C 3-9 Heterocycloalkyl may be mono- or poly-substituted by: 1-3 Alkyl, halogen, -C 1-3 haloalkyl, -OH, oxo or cyano;

[0024] R a and R b or together with the nitrogen atom to which they are attached, form an unsubstituted or substituted group or -NR c R d Substituted 4-9 membered heterocycloalkyl, T is -C 1-4 Alkyl, -C 1-3 Alkyl-substituted-C2-4 Alkyl, -C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, -C 1-3 Haloalkyl, -(CH2) n -NR c R d ;

[0025] R c and R d are independently hydrogen, -C 1-3 Alkyl, -C 1-4 Alkylene -OH, -C 2-4 Alkylene -OCH3, -(CH2)2-OC 1-3 alkyl;

[0026] R 3 -NH-C 4-8 Cycloalkyl, -NH-C 3-8 Heterocycloalkyl, -NH-CH2-C 3-8 Heterocycloalkyl or -OC 1-4 Alkyl; R 3 Preferably -OC 1-4 Alkyl, -NH-CH2-C 4-5 Heterocycloalkyl; T is -C 1-4 Alkyl; T 1 are -O, -S, -NH, -N-CH3;

[0027] R 4 is phenyl, heteroaryl, -NH-phenyl, wherein the phenyl group may be R 2 and / or R k Substituted, the heteroaryl group may be R 2 Replacement; R k Halogen or -OC 1-4 Alkyl; R 4 Preferably X1 is carbon or nitrogen;

[0028] R 6 is phenyl or 4-6 membered heterocycloalkyl, wherein the phenyl or 4-6 membered heterocycloalkyl is mono- or di-substituted by the following groups: halogen, -OH, -C 1-3 Alkyl, -C(CH3)2-OH or cyano; R 6 Preferably

[0029] R 7 Hydrogen, -C 1-3 Alkyl, -(CH2) n -OH, C 1-3 Haloalkyl, -(CH2)n -OC 1-3 Alkyl, -(CH2) n -OC(O)-C 1-3 Alkyl; R 7 Preferably, it is hydrogen, -(CH2) n -OH, -(CH2) n -F, -(CH2) n -OC 1-3 alkyl;

[0030] n is 1 or 2.

[0031] The compound provided by the present invention has a specific structure as follows:

[0032] N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0033] N-(6-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)imidazo[1,2-b]pyridazin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0034] N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0035] 5-(3,5-difluorophenyl)-3-(2'-(4-methylpiperazin-1-yl)-[2,4'-bipyridyl]-6-yl)-1H-indazole;

[0036] 6-(5-(3,5-difluorophenyl)-1H-indazol-3-yl)-N-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)pyridin-2-amine;

[0037] N-(5-(1-(3,5-difluorophenyl)cyclopropyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzoamide;

[0038] N-(5-((3,5-difluorophenyl)difluoromethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0039] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((4-methylpiperazin-1-yl)sulfonyl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0040] N-(5-(3,5-difluorobenzyl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0041] N-(5-(1-(3,5-difluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0042] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(oxetan-3-ylamino)benzamide;

[0043] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0044] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0045] (R)-N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0046] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(7-methyl-2,7-diazaspiro[3.5]nonan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0047] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(6-methyl-2,6-diazaspiro[3.3]heptan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0048] N-(5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0049] N-(6-(3,5-difluorobenzyl)imidazo[1,2-b]pyridazin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydropyran-4-yl)amino)-benzamide;

[0050] N-(5-((4-(2-hydroxypropan-2-yl)piperidin-1-yl)methyl)-1H-indazol-3-yl)-4-(4-methylpiperidin-1-yl)-2-((tetrahydropyran-4-yl)amino)benzamide;

[0051] N-(5-(3,5-difluorophenyl)-1H-indazol-3-yl)-4-(5-(dimethylamino)hexahydrocyclopenta[c]pyrrol-2(1H)-yl)-2((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0052] N-(5-(1-(3,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0053] 5-(3,5-difluorophenyl)-N-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)-1H-indazole-3-carboxamide;

[0054] N-(5-(1-(3,5-difluorophenyl)-2-methoxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0055] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-methyl-7-((tetrahydro-2H-pyran-4-yl)amino)-1,2,3,4-tetrahydroisoquinoline-6-carboxamide;

[0056] 4-((1R,3S,5s,7s)-5-aminoadamantan-2-yl)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide hydrochloride;

[0057] 4-(5-amino-adamantan-2-yl)-N-(5-(3,5-difluorobenzyl)-1H-indol-3-yl)-2-(piperidin-4-ylamino)benzamide;

[0058] (R)-N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0059] N-(5-(1-(2,5-difluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0060] Ethyl 2-(3,5-difluorophenyl)-2-(3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamido)-1H-indazol-5-yl)acetate;

[0061] N-(5-(1-(3,5-difluorophenyl)ethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0062] N-(5-(3-fluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0063] N-(5-(1-(3-fluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0064] N-(5-(1-(3-fluorophenyl)ethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0065] N-(5-(1-(2,5-difluorophenyl)ethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0066] N-(5-(1-(2,5-difluorophenyl)-2-methoxyethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0067] 2-(3-Fluorophenyl)-2-(3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)-1-hydro-indazol-5-yl)ethyl isobutyl ester;

[0068] N-(5-(3-fluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(5-methyl-2,5-diazabicyclo[2.2.1]hept-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0069] N-(5-(1-(3-fluorophenyl)-2-methoxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0070] N-(5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(5-methyl-2,5-diazabicyclo[2.2.1]hept-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0071] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(3-(dimethylamino)azetidin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0072] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(3-((2-methoxyethyl)(methyl)amino)azetidin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0073] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0074] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0075] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(dimethylamino)ethyl)thio)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0076] N-(5-(3-fluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0077] N-(5-(3-fluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0078] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((oxetan-3-ylmethyl)amino)benzamide;

[0079] N-(5-(3-fluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((oxetan-3-ylmethyl)amino)benzamide;

[0080] N-(5-(2,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((oxetan-3-ylmethyl)amino)benzamide;

[0081] N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0082] N-(5-(1-(3-fluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0083] N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-ethylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0084] N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-morpholinopiperidin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0085] N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0086] N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-cyclopropylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0087] N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-trifluoromethylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0088] N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0089] N-(5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-((1R,4R)-5-methyl-2,5-diazabicyclo[2.2.10.1]heptan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0090] N-(5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-((1s,4s)-5-methyl-2,5-diazabicyclo[2.2.10.1]heptan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0091] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-(2-(dimethylamino)ethyl)-1-carbonyl-6-((tetrahydro-2H-pyran-4-yl)amino)isoindoline-5-carboxamide;

[0092] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(N-methyl-2-morpholinoacetylamino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0093] N-(5-(1-(3,5-difluorophenyl)-2,2,2-trifluoro-ethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0094] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0095] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(3-fluoropyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0096] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(pyrrolidin-1-yl)ethyl)thio)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0097] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)--4-((2-(3,3-difluoropyrrolidin-1-yl)-ethyl)-(ethyl)-amino)--2-((tetrahydro-2H-pyran-4-yl)amino)-benzamide;

[0098] N-(5-(3,5-difluorobenzyl)--1H-indazol-3-yl)--4-((2-(3,4-difluoropyrrolidin-1-yl)-ethyl)(ethyl)-amino)--2-((tetrahydro-2H-pyran-4-yl)-amino)-benzamide;

[0099] N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(3,4-dimethylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0100] N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(3-methyl-4-ethylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0101] N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(2,4-dimethylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0102] N-(5-(3,5-difluorobenzyl(-1H-indazol-3-yl)-4-((2-(3,3,4,4-tetrafluoropyrrolidin-1-yl)-ethyl)-(ethyl)-amino)-2-((tetrahydro-2H-pyran-4-yl)-amino)-benzamide;

[0103] N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)-4-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)benzamide;

[0104] N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(2-methyl-4-ethylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0105] 4-(Cyclopropyl(2-(pyrrolidin-1-yl)-ethyl)-amino)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)--2-((tetrahydro-2H-pyran-4-yl)-amino)-benzamide;

[0106] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)--4-((2-(1,4-dioxo-7-azaspiro[4.4]nonan-1-yl)ethyl)-(ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)-amino)-benzamide;

[0107] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(3-oxopyrrolidin-1-yl)ethyl)(ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0108] N-(5-(2,5-difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0109] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(azetidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0110] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(pyrrolidin-1-yl)ethyl)sulfinyl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0111] N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methyl-4,7-diazaspiro[2.5]octan-7-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0112] N-(5-(1-(2,5-difluorophenyl(-2-fluoroethyl)-1H-indazol-3-yl)-4-(3,5-dimethylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0113] N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-cyanopyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0114] (R)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-hydroxypyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide;

[0115] (S)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-hydroxypyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide.

[0116] The present invention provides a method for preparing a compound represented by general formula II. The compound represented by general formula II is prepared by the following scheme:

[0117] Scheme 1, when Y 1 When it is carbon, Y 3 Nitrogen, Y 2 For carbon, Y 4 For carbon, Y 5 R is carbon; 1 For-CF2-R 6 The synthetic route is summarized as follows:

[0118]

[0119] The oxidizing agent is preferably a Dess-Martin oxidant, PCC, PDC or other oxidizing agents, the fluorinating agent is preferably a fluorinating agent such as BAST, DAST or other fluorinating agents, and the amide forming method is preferably a carboxylic acid reacting an acyl chloride with an amine to form an amide, or a carboxylic acid is activated with a condensing agent such as 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) / 1-hydroxybenzotriazole (HOBT), cyclohexylcarbodiimide (DCC), 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) to form an amide with an amine;

[0120] Option 2, when Y 1 When it is carbon, Y 3 Nitrogen, Y 2 For carbon, Y 4 For carbon, Y 5 R is carbon; 1 for

[0121] The synthetic route is summarized as follows:

[0122]

[0123] The oxidizing agent is preferably a Dess-Martin oxidant, PCC, PDC or other oxidizing agents; the three-membered ring forming method is preferably diethylzinc / diiodomethane, trimethylsulfoxide iodide or trimethylsulfoxide bromide / sodium hydrogen; the boronating agent is preferably borane, 9-BBN or the like; the oxidizing agent is preferably hydrogen peroxide; the fluorinating agent is preferably a fluorinating agent such as BAST or DAST; the amide forming method is preferably a carboxylic acid reacting an acyl chloride with an amine to form an amide, or a carboxylic acid is activated with a condensing agent such as 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) / 1-hydroxybenzotriazole (HOBT), cyclohexylcarbodiimide (DCC), 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) to form an amide with an amine;

[0124] Option 3, when Y 1 When it is carbon, Y 2 For carbon, Y 3 Nitrogen, Y 4 For carbon, Y 5 is nitrogen; or Y 1 When Y is nitrogen, 4 Nitrogen, Y 2 For carbon, Y 3 For carbon, Y 5 is carbon; or Y 1 When Y is nitrogen, 2 Nitrogen, Y 3 For carbon, Y 4 For carbon, Y 5 R is carbon; 1 -CH2-R 6 The synthetic route is summarized as follows:

[0125]

[0126] Wherein the halogen is preferably bromine, iodine, N-bromosuccinimide, N-iodosuccinimide, etc., and the coupling reaction is preferably Stille reaction;

[0127] Scheme 4, when Y 1 When it is carbon, Y 2 For carbon, Y 3 Nitrogen, Y 4 For carbon, Y 5 is carbon; or Y 1 for

[0128]

[0129] The nitrating agent is preferably nitric acid, potassium nitrate, sodium nitrate, etc.; the reducing agent is preferably iron powder, zinc powder, stannous chloride, hydrogen / palladium carbon, hydrogen / nickel, etc.; the amide forming method is preferably a method in which carboxylic acid is reacted with acyl chloride and amine to form amide, or carboxylic acid is activated with a condensing agent such as 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) / 1-hydroxybenzotriazole (HOBT), cyclohexylcarbodiimide (DCC), 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) to form amide with amine;

[0130] Scheme 5, when R 0 for R 1 -CH2-R 6 ; The synthetic route is summarized as follows:

[0131]

[0132] Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof.

[0133] Suitable acid addition salts are formed from acids which form non-toxic salts, examples of which include, but are not limited to, hydrochlorides, sulfates / bisulfates, nitrates, phosphates / hydrogenphosphates / dihydrogenphosphates, hydrobromides, hydroiodides, acetates, lactates, methanesulfonates, citrates, malates, maleates, fumarates, tartrates, salicylates, stearates, and the like.

[0134] Suitable base addition salts are formed from bases which form non-toxic salts, for example as alkali metal salts, alkaline earth metal salts or as ammonium salts, examples include, but are not limited to, sodium salts, potassium salts, calcium salts, magnesium salts, or salts with ammonia or organic amines such as ethylamine, ethanolamine, triethanolamine or amino acids.

[0135] For a review of suitable salts, see Stahl, PH and Wermuth, CG, "Handbook of Pharmaceutical Salts: Properties, Selection, and Use, 2 nd Revised Edition” (Wiley-VCH, 2011).

[0136] If the compounds of the formula (II) contain both acidic and basic groups in the molecule, the invention also includes inner salts or betaines (zwitterions) in addition to the salt forms mentioned.

[0137] Methods for preparing pharmaceutically acceptable salts of the compounds of the present invention are known to those skilled in the art, for example, by contacting these compounds with an organic or inorganic acid or base in a solvent or dispersant, or by anion exchange or cation exchange with other salts.

[0138] The compound of formula (II) may exist in a crystalline or amorphous form. In addition, some crystalline forms of the compound of formula (II) may exist in polymorphic forms, which are included within the scope of the present invention. Many conventional analytical techniques may be used including but not limited to single crystal X-ray powder diffraction (XRPD) patterns, infrared (IR) spectra, Raman spectroscopy, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) and solid state nuclear magnetic resonance (ssNMR) characterizations to distinguish the polymorphic forms of the compound.

[0139] The compounds of the present invention also include tautomeric forms. Tautomeric forms are produced by the exchange of a single bond with an adjacent double bond and the concomitant migration of a proton. Tautomeric forms include proton transfer tautomers, which are isomeric protonation states with the same empirical formula and total charge. Examples of proton transfer tautomers include keto-enol pairing, amide-imidic acid pairing, lactam-lactim pairing, enamine-imine pairing, and cyclic forms in which protons can occupy two or more positions of a heterocyclic system, for example, 1H- and 3H-imidazoles, 1H-, 2H- and 4H-1,2,4 triazoles, 1H- and 2H-isoindoles, and 1H- and 2H-pyrazoles. Tautomeric forms can be in equilibrium or spatially fixed to one form by appropriate substitution.

[0140] The present invention further relates to a compound represented by general formula (II) or its tautomers, mesomers, racemates, enantiomers, diastereomers, and mixtures thereof, and pharmaceutically acceptable salts thereof or pharmaceutical compositions containing the same, which are further used in combination with one or more other agents for regulating the immune system of mammals, anticancer agents or anti-inflammatory agents.

[0141] The present invention further relates to a compound represented by general formula (II) or its tautomers, mesomers, racemates, enantiomers, diastereomers, and mixtures thereof, and pharmaceutically acceptable salts thereof or pharmaceutical compositions comprising the same, which are used to treat or prevent the following cancers, including solid tumors (such as prostate cancer, kidney cancer, liver cancer, pancreatic cancer, gastric cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, melanoma, etc.), blood cancers (such as lymphoma, leukemia, etc.), skin cancers (such as cutaneous T-cell lymphoma, cutaneous B-cell lymphoma), etc.

[0142] The present invention also relates to a method for treating or preventing cancer, which comprises administering to a patient in need thereof a therapeutically effective amount of a compound represented by general formula (II) or its tautomers, mesomers, racemates, enantiomers, diastereomers, and mixtures thereof, and pharmaceutically acceptable salts thereof, or a pharmaceutical composition comprising the same, wherein the cancer comprises, for example, solid tumors (such as prostate cancer, kidney cancer, liver cancer, pancreatic cancer, gastric cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, melanoma, etc.), blood cancer (such as lymphoma, leukemia, etc.), skin cancer (such as cutaneous T-cell lymphoma, cutaneous B-cell lymphoma), etc.

[0143] The compositions of the present invention may be formulated in a conventional manner using one or more pharmaceutically acceptable carriers. Thus, the active compounds of the present invention may be formulated into dosage forms for oral, buccal, intranasal, parenteral (e.g., intravenous, intramuscular or subcutaneous) or rectal administration, or dosage forms suitable for administration by inhalation or insufflation. The compounds of the present invention may also be formulated into sustained release dosage forms.

[0144] Depending on the purpose of treatment, the pharmaceutical composition can be prepared into various types of dosage unit forms, such as tablets, pills, powders, liquids, suspensions, emulsions, granules, capsules, suppositories and injections (solutions and suspensions), or dosage forms suitable for administration by inhalation or insufflation.

[0145] In order to shape the pharmaceutical composition into tablet form, any excipient known and widely used in the art may be used. For example, carriers such as lactose, white sugar, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose and silicic acid; binders such as water, ethanol, propanol, ordinary syrup, glucose solution, starch solution, gelatin solution, carboxymethyl cellulose, shellac, methyl cellulose and potassium phosphate, polyvinyl pyrrolidone, etc.; disintegrants such as dry starch, sodium alginate, agar powder and kelp powder, sodium bicarbonate, calcium carbonate, fatty acid esters of polyethylene sorbitan, sodium lauryl sulfate, monoglyceride of stearic acid, starch and lactose, etc.; disintegration inhibitors such as white sugar, glyceryl tristearate, coconut oil and hydrogenated oil; adsorption promoters such as quaternary ammonium base and sodium lauryl sulfate, etc.; wetting agents such as glycerol and starch, etc.; adsorbents such as starch, lactose, kaolin, bentonite and colloidal silicic acid, etc.; and lubricants such as purified talc, stearate, boric acid powder and polyethylene glycol, etc. Ordinary coating materials can be selected as needed to make sugar-coated tablets, gelatin-film-coated tablets, enteric-coated tablets, film-coated tablets, double-layer film tablets and multi-layer tablets.

[0146] In order to shape the pharmaceutical composition in the form of pills, any excipient known and widely used in the art can be used, for example, carriers such as lactose, starch, coconut oil, hardened vegetable oil, kaolin and talc, etc.; binders such as gum arabic powder, tragacanth powder, gelatin and ethanol, etc.; disintegrants such as agar and kelp powder, etc.

[0147] In order to shape the pharmaceutical composition into a suppository form, any excipient known and widely used in the art may be used, for example, polyethylene glycol, coconut oil, higher alcohols, esters of higher alcohols, gelatin and semi-synthetic glycerides and the like.

[0148] In order to prepare the pharmaceutical composition in the form of injection, the solution or suspension can be sterilized (preferably by adding an appropriate amount of sodium chloride, glucose or glycerol, etc.) to prepare an injection with an osmotic pressure equal to that of blood. When preparing the injection, any commonly used carrier in the art can also be used. For example, water, ethanol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol and fatty acid esters of polyethylene sorbitan, etc. In addition, common solvents, buffers and analgesics, etc. can also be added.

[0149] For intranasal administration or administration by inhalation, the active compound of the invention is suitably released from a pump spray container which is received or squeezed or aspirated in the form of a solution or suspension, or released from a pressurized container or atomizer in the form of an aerosol, using a suitable propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve for metered release. The pressurized container or atomizer can contain a solution or suspension of the active compound. Capsules or cartridges (e.g., made of gelatin) for use in an inhaler or insufflator can be formulated to contain a powder mixture of the compound of the invention and a suitable powder base, such as lactose or starch.

[0150] Unless stated otherwise, the terms used in the specification and claims have the following meanings.

[0151] The term "stereoisomer" refers to an isomer formed due to at least one asymmetric center. In compounds with one or more (e.g., one, two, three, or four) asymmetric centers, racemates, racemic mixtures, mesomorphs, single enantiomers, diastereomeric mixtures, and individual diastereomers may occur. Specific individual molecules may also exist as geometric isomers (cis / trans).

[0152] The term "solvate" refers to a compound form that is physically bound to a solvent, usually by a solvolysis reaction. This physical binding includes hydrogen bonding. Conventional solvents include water, ethanol, methanol, acetic acid, etc. The compound of formula (II) can be prepared in a crystalline form and can be in the form of a solvate (e.g., a hydrated form). Suitable solvates include pharmaceutically acceptable solvates (e.g., hydrates), and further include stoichiometric solvates and non-stoichiometric solvates. In some cases, for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate will be able to dissociate. "Solvate" encompasses solution phase and dissociable solvates. Representative solvates include hydrates, ethanolates, and methanolates, etc.

[0153] The term "prodrug" refers to a derivative that is converted into the compound of the present invention by reaction with an enzyme, gastric acid, etc. under physiological conditions in vivo, for example, by oxidation, reduction, hydrolysis, etc., each catalyzed by an enzyme.

[0154] The term "metabolites" refers to all molecules derived from any compound of the invention in a cell or organism, preferably a human.

[0155] The term "isotopic derivative" refers to a compound that contains isotopes in unnatural proportions at one or more atoms of the compound. 2 H or D), tritium ( 3 H or T), carbon-13 ( 13 C), nitrogen-15 ( 15 N), oxygen-18 ( 18 O) etc.

[0156] The term "pharmaceutical composition" refers to a mixture containing one or more compounds of the present invention or their physiologically / pharmaceutically acceptable salts or prodrugs and other chemical components, such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitate the absorption of the active ingredient, and thus exert biological activity.

[0157] The carrier includes all pharmaceutical preparations that can be used for injection and non-injection administration in the pharmaceutical field, such as diluents, wetting agents, fillers, adhesives, lubricants, disintegrants, absorption promoters, surfactants, retardants, adsorbents, suspending agents, flocculants, deflocculants, emulsifiers, common bases, solubilizers, cosolvents, latent solvents, preservatives, flavoring agents, colorants, antioxidants, buffers, antibacterial agents, isotonicity regulators, pH regulators, metal ion complexing agents, hardeners, thickeners, etc.

[0158] The novel RTK inhibitors provided in the present application have the potential to cause cancer cells that rely on RTK activity disorders to die without damaging normal tissues. In addition, the TRK family protein tyrosine kinase inhibitors provided in the present application can be used to treat pain, inflammation, cancer and certain infectious diseases. The inventors unexpectedly found that the RTK inhibitors provided in the present application can effectively inhibit the activity of tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK), and proto-oncogene tyrosine kinase (ROS1), and have a strong inhibitory effect on the proliferation of tumor cells and cancer cells and tumor growth, and the inhibitory activity is better than that of Entrectinib in the prior art. In addition, the compounds of the present invention also have good metabolic characteristics and good bioavailability. DETAILED DESCRIPTION

[0159] In order to make the purpose, technical scheme, and advantages of the present invention clearer, the present invention is further described below in conjunction with embodiments, but these embodiments do not limit the scope of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention belong to the scope of protection of the present invention. The experimental methods in the embodiments of the present invention that do not specify specific conditions are usually carried out under conventional conditions, or under conditions recommended by raw materials or commodity manufacturers. Reagents that do not specify specific sources are conventional reagents purchased from the market.

[0160] Intermediate preparation method example:

[0161] 1. Preparation of 4-(4-methylpiperazine-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydropyran-4-yl)acetamido)benzoic acid I-1

[0162]

[0163] The first step is the preparation of tert-butyl 4-fluoro-2-nitro-benzoate I-1b

[0164] Compound 4-fluoro-2-nitro-benzoic acid I-1a (10 g, 54 mmol), Boc anhydride (23.6 g, 108 mmol) and DMAP (2 g, 16.2 mmol) were dissolved in a mixed solution of DCM (100 mL) and tert-butanol (100 mL), stirred overnight at room temperature, concentrated, diluted with 500 mL of ethyl acetate, washed with 1N HCl (500 mL*1), washed with water (500 mL*1), washed with saturated brine (500 mL*1), dried over anhydrous Na2SO4, filtered, concentrated to obtain the title compound I-1b (13 g, 54 mmol), with a yield of 100%.

[0165] Step 2 Preparation of tert-butyl 4-(4-methylpiperazine-1-yl)-2-nitrobenzoate I-1c

[0166] Compound 2 (13 g, 54 mmol) was added to 1-methylpiperazine (17 mL) and reacted at room temperature overnight. 400 mL of water was added and stirred for 1 hour, filtered, washed with water, and dried to obtain the title compound I-1c (16 g, 49.8 mmol) with a yield of 92.4%.

[0167] MS m / z(ESI):322[M+H] + .

[0168] Step 3 Preparation of tert-butyl 4-(4-methylpiperazine-1-yl)-2-aminobenzoate I-1d

[0169] Compound I-1c (16 g, 49.8 mmol) was dissolved in methanol (200 mL), and water (200 mL), ammonium chloride (10.7 g, 200 mmol) and zinc powder (35 g, 538 mmol) were added in sequence, and the mixture was reacted at room temperature overnight. 800 mL of water was added for dilution, and ethyl acetate (300 mL╳2) was added for extraction, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by Flash column chromatography (PE / EA, 0-60%) to obtain the title compound I-1d (5.9 g, 20.3 mmol) with a yield of 40.7%.

[0170] MS m / z(ESI):292[M+H] + .

[0171] Step 4 Preparation of tert-butyl 4-(4-methylpiperazine-1-yl)-2-(tetrahydropyran-4-yl)aminobenzoate I-1e

[0172] Compound I-1d (5 g, 17.2 mmol) was dissolved in DCM (100 mL), tetrahydropyran-4-one (5 g, 50 mmol) and acetic acid (5 mL) were added, and the mixture was stirred at room temperature for 30 minutes, and then sodium triacetoxyborohydride (15 g, 71.1 mmol) was added, and the mixture was stirred at room temperature overnight. The mixture was concentrated and purified by flash column chromatography (DCM / MeOH, 0-10%) to obtain the title compound I-1e (6 g, 16 mmol) with a yield of 93.1%.

[0173] MS m / z(ESI):376[M+H] + .

[0174] Step 5 Preparation of tert-butyl 4-(4-methylpiperazine-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydropyran-4-yl)acetamido)benzoate I-1f

[0175] Compound I-1e (5 g, 13.3 mmol) was dissolved in DCM (50 mL), and triethylamine (2.3 g, 23 mmol) and trifluoroacetic anhydride (4.2 g, 20 mmol) were added. After stirring in an ice bath for 20 minutes, 30 mL of water was added dropwise. The liquids were separated, and the organic phase was washed with saturated brine (30 mL), dried over anhydrous Na2SO4, filtered, concentrated, and purified by Flash column chromatography (DCM / MeOH, 0-5%) to obtain the title compound I-1f (5.3 g, 11.2 mmol) with a yield of 84.4%.

[0176] MS m / z(ESI):472[M+H] + .

[0177] Step 6 Preparation of 4-(4-methylpiperazine-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydropyran-4-yl)acetamido)benzoic acid I-1

[0178] Compound I-1f (5 g, 10.6 mmol) was dissolved in DCM (100 mL), trifluoroacetic acid (25 mL) was added, and the mixture was reacted at room temperature for 2 hours, concentrated, methyl tert-butyl ether (200 mL) was added, and stirring was continued for 15 minutes, filtered, washed with methyl tert-butyl ether, and dried to obtain the title compound I-1 (4.6 g, 11.1 mmol), with a yield of 97.3%.

[0179] 1 H NMR (400MHz, DMSO-d6) δ12.77(s,1H),10.03(s,1H),7.87(d,J=8.9Hz,1H),7.08(dd,J1=8.9,J2=2.6Hz,1H),6.94(d,J=2.6Hz,1 H),4.44(m,1H),4.06(s,2H),3.79(m,2H),3.59–3.04(m,8H),2.83(s,3H),1.92(m,1H),1.56(m,1H),1.47(m,1H),1.01(m,1H).

[0180] MS m / z(ESI):416[M+H]+.

[0181] 2. Preparation of 3-amino-5-((3,5-difluorophenyl)methyl)-1H-indazole I-2

[0182]

[0183] The first step is the preparation of (3-cyano-4-fluorophenyl)-(3,5-difluorophenyl)methanol I-2c

[0184] 3,5-Difluorobromobenzene I-2b (3.8 g, 20 mmol) was added dropwise in batches to a refluxing suspension of dry tetrahydrofuran (30 mL) containing magnesium chips (0.48 g, 20 mmol). After reflux for 3 hours, it was cooled to room temperature and added dropwise to a solution of I-2a (3 g, 20 mmol) in dry tetrahydrofuran (30 mL). The temperature did not exceed -15 °C. After the addition was completed, it was stirred for 30 minutes, raised to 0 °C, and saturated ammonium chloride solution was added to quench the reaction. Part of the tetrahydrofuran was evaporated under reduced pressure, extracted with ethyl acetate, dried, concentrated, and column chromatographed to obtain the product I-2c (3.6 g, 13.7 mmol) with a yield of 64%.

[0185] MS m / z(ESI):264[M+H] + .

[0186] Step 2 Preparation of (3-cyano-4-fluorophenyl)-(3,5-difluorophenyl)methane I-2d

[0187] Compound I-2c (5.26 g, 20 mmol) was dissolved in acetonitrile (100 mL), sodium iodide (30 g, 200 mmol) was added and heated to 65 ° C. Trimethylsilyl chloride (21.7 g, 200 mmol) was added in batches within 6 hours. The mixture was cooled to room temperature, sodium thiosulfate solution was added and stirred until colorless, and extracted with ethyl acetate. The white solid product I-2d (3.2 g, 13 mmol) was dried and concentrated, with a yield of 65%.

[0188] MS m / z(ESI):248[M+H] + .

[0189] Step 3 Preparation of 3-amino-5-((3,5-difluorophenyl)methyl)-1H-indazole I-2

[0190] Compound I-2d (3 g, 12 mmol) and 80% hydrazine hydrate (3.8 g, 60 mmol) were added to N-methylpyrrolidone (15 mL), heated at 150 ° C for 4 hours, cooled to room temperature, added 25 mL of water, stirred until solid precipitated, and the white solid product I-2 (2.2 g, 8.5 mmol) was filtered, with a yield of 71%.

[0191] MS m / z(ESI):260[M+H] + .

[0192] 1 H NMR (400MHz, DMSO-d6) δ11.28(s,1H),7.49(s,1H),7.17–7.05(m,2H),6.99(tt,J=9.4,2.4Hz,1H),6.94–6.85(m,2H),5.21(s,2H),3.96(s,2H).

[0193] Product preparation method example:

[0194] Example 1

[0195] Preparation of N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 1

[0196]

[0197] The first step is the preparation of 5-chloro-3-nitropyrazolo[1,5-a]pyrimidine 1b

[0198] 5-Chloropyrazolo[1,5-a]pyrimidine 1a (2 g, 13 mmol) was dissolved in H2SO4 (8 mL), KNO3 (2 g, 19.6 mmol) was added under ice bath, and the mixture was slowly heated to room temperature under stirring. The mixture was slowly poured into ice water, and a yellow solid precipitated. The solid was filtered, collected and dried to obtain 5-chloro-3-nitropyrazolo[1,5-a]pyrimidine 1b (2 g, 10.1 mmol), with a yield of 77%.

[0199] 1 H NMR (400MHz, DMSO-d6) δ9.43 (d, J = 7.2 Hz, 1H), 9.07 (s, 1H), 7.61 (d, J = 7.2 Hz, 1H).

[0200] Step 2 Preparation of 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-3-nitropyrazolo[1,5-a]pyrimidine 1d

[0201] 2-(2,5-difluorophenyl)pyrrolidine 1c (600 mg, 3.27 mmol) and triethylamine (990 mg, 9.81 mmol) were dissolved in THF, and a THF solution of 5-chloro-3-nitropyrazolo[1,5-a]pyrimidine 1b (650 mg, 3.27 mmol) was added dropwise at room temperature. After the addition was completed, stirring was continued at room temperature for 2 hours. Quenching was added, and the mixture was extracted with ethyl acetate three times. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography (PE:EA=1:1) to obtain 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-3-nitropyrazolo[1,5-a]pyrimidine 1d (0.9 g, 2.6 mmol) as a yellow solid with a yield of 79%.

[0202] MS m / z(ESI):346[M+H] + .

[0203] Step 3 Preparation of 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-amine 1e

[0204] 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-3-nitropyrazolo[1,5-a]pyrimidine 1d (400 mg, 1.15 mmol) and zinc powder (376 mg, 5.79 mmol) were dispersed in THF (5 mL), and 3 mL of saturated aqueous ammonium chloride solution was added. The mixture was stirred at room temperature for 1 hour, filtered, and the filtrate was extracted twice with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography (DCM:MeOH=10:1) to obtain 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidine-3-amine 1e (200 mg, 0.63 mmol) as a light yellow solid in a yield of 55%.

[0205] MS m / z(ESI):316[M+H] + .

[0206] Step 4 Preparation of 4-(4-methylpiperazine-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetylamino)benzoyl chloride 1f

[0207] 4-(4-Methylpiperazine-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoic acid I-1 (150 mg, 0.36 mmol) was dissolved in DCM, oxalyl chloride (229 mg, 1.8 mmol) and a drop of DMF were added, and the mixture was stirred at reflux for 1 hour. The reaction solution was concentrated and the crude product 1f was directly used in the next reaction.

[0208] Step 5 Preparation of 1 g of N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazine-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide

[0209] 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-amine 1e (100 mg, 0.31 mmol) and triethylamine (93 mg, 0.92 mmol) were dissolved in DCM, and a DCM solution of 4-(4-methylpiperazine-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoyl chloride 1f (165 mg, 0.38 mmol) was added dropwise at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated and 1 g of the crude product was directly used in the next reaction.

[0210] Step 6 Preparation of N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 1

[0211] Dissolve 1 g (150 mg, 0.21 mmol) of N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide in methanol and add 5N K2CO3 (4 mL), the reaction mixture was stirred at room temperature for 2 hours, the reaction solution was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography (DCM: MeOH = 10: 1) to give N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 1 (60 mg, 0.097 mmol) as an apricot solid, with a yield of 46%.

[0212] MS m / z(ESI):617[M+H] +

[0213] 1H NMR(400MHz,DMSO-d6)δ9.03(br,1H),8.49(br,1H),8.15(br,1H),7.92(s,1H),7.59(br,1H), 7.31–7.02(m,3H),6.83(br,1H),6.20(dd,J=8.9,2.2Hz,1H),6.10(d,J=2.3Hz,1H),5.34(br, 1H),3.94–3.85(m,1H),3.78(dt,J=11.6,3.9Hz,2H),3.68-3.53(m,2H),3.49–3.40(m,2H),3. 21(t,J=5.1Hz,4H),2.40(t,J=5.0Hz,4H),2.19(s,3H),2.02-1.81(m,6H),1.37–1.25(m,2H).

[0214] Example 2

[0215] Preparation of N-(6-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)imidazo[1,2-b]pyridazin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 2

[0216]

[0217] The same method as in Example 1 was used to prepare the compound, except that 6-chloro-3-nitroimidazo[1,2-b]pyridazine was used as the starting material instead of 1b.

[0218] MS m / z(ESI):617[M+H]+

[0219] 1 H NMR(400MHz,DMSO-d6)δ9.23(s,1H),7.59(br,4H),7.21-7.05(m,3H),6.91–6.8 6(m,1H),6.16(br,1H),6.07(br,1H),5.26(d,J=8.1Hz,1H),3.96–3.85(s,1H),3 .81(d,J=11.4Hz,2H),3.68–3.58(m,2H),3.44(t,J=10.7Hz,2H),3.20(t,J=5.1H z,4H),2.41(t,J=5.1Hz,4H),2.19(s,3H),2.01–1.79(m,6H),1.52-1.30(m,2H).

[0220] Example 3

[0221] Preparation of N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 3

[0222]

[0223] The first step is the preparation of tert-butyl-3-[di(tert-butoxycarbonyl)amino]-5-bromo-1H-indazole-1-carboxylate 3b

[0224] Compound 3a (1.0 g, 4.7 mmol), (Boc)2O (3.1 g, 14.1 mmol) and DMAP (0.57 g, 4.7 mmol) were added to DCM (20 ml) and reacted at room temperature overnight. The solvent was evaporated under reduced pressure, EA (50 ml) was added, and the mixture was washed with satNH4Cl (15 ml×2) and brine (20 ml) in sequence, dried over anhydrous sodium sulfate, concentrated, and purified by flash column (petroleum ether to petroleum ether / ethyl acetate = 20:1) to obtain white foam solid compound 3b (1.5 g) with a yield of 63%.

[0225] MS m / z(ESI):534[M+Na] +

[0226] Step 2 Preparation of 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-1-(N-tert-butyloxycarbonyl)-indazole-3-(di(tert-butyloxycarbonyl))amine 3c

[0227] Compound 3b (1.3 g, 2.5 mmol), compound 1c (0.38 g, 2.1 mmol) and Cs2CO3 (1.4 g, 4.2 mmol) were added to dioxane (30 ml), and Pd2(dba)3 (0.19 g, 0.21 mmol) and Ruphos (0.20 g, 0.42 mmol) were added in sequence, and the mixture was reacted at 92°C for 4 h. After cooling to room temperature, water (20 ml) was added, and the mixture was extracted with ethyl acetate (50 ml×2). The organic layers were combined, washed with brine (20 ml), dried over anhydrous sodium sulfate, concentrated, and purified by flash column (petroleum ether to petroleum ether / ethyl acetate = 5:1) to obtain a white foamy solid, compound 3c (0.8 g), with a yield of 62%.

[0228] MS m / z(ESI):615[M+1] +

[0229] Step 3 Preparation of 5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-amine 3d

[0230] Compound 3c (0.8 g, 1.3 mmol) was added to 4M HCl / dioxane (20 ml) and reacted at room temperature overnight. It was evaporated to dryness under reduced pressure to obtain a yellow solid, compound 3d (0.41 g), with a yield of 100%.

[0231] MS m / z(ESI):315[M+1] +

[0232] Step 4 Preparation of N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 3e

[0233] Compound 3d (50 mg, 0.16 mmol) and DIEA (83 mg, 0.64 mmol) were added to anhydrous THF (5 ml), cooled to -20°C, 1f (100 mg) was dissolved in anhydrous THF (1 ml), 0.7 ml was taken with a syringe and added dropwise to the above solution, and the reaction was continued for 2 h. Water (10 ml) was added, and the mixture was extracted with ethyl acetate (15 ml×2), the organic layers were combined, washed with brine (15 ml), dried over anhydrous sodium sulfate, concentrated, and purified by flash column (DCM~DCM / MeOH=6:1) to obtain a yellow solid, compound 3e (30 mg), with a yield of 26%.

[0234] Step 5 Preparation of N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 3

[0235] Compound 3e (30 mg, 0.042 mmol) was added to MeOH (6 ml) and TEA (ml), refluxed for 2 h, the solvent was evaporated under reduced pressure, ethyl acetate (20 ml) was added, washed with water (8 ml) and brine (10 ml) in sequence, dried over anhydrous sodium sulfate, concentrated, and purified by flash column (DCM ~ DCM / MeOH = 6:1) to give a nearly white solid, the final product 3 (15 mg).

[0236] MS m / z(ESI):616[M+1] +

[0237] 1H NMR (400MHz, DMSO-d6) δ12.32(s,1H),9.94(s,1H),8.28(d,J=7.7Hz,1H),7.76(d,J=9.0Hz,1H),7.3 1–7.19(m,2H),7.14–7.04(m,1H),6.87(m,1H),6.66(dd,J=9.1,2.3Hz,1H),6.49(d,J=2.2Hz,1H),6 .22(dd,J=9.0,2.3Hz,1H),6.14(d,J=2.3Hz,1H),4.91(d,J=8.3Hz,1H),3.83(m,2H),3.71(m,2H),3 .56–3.45(m,2H),3.27(m,5H),2.48–2.36(m,4H),2.24(s,3H),2.05–1.80(m,5H),1.42–1.25(m,3H).

[0238] Example 4

[0239] Preparation of 5-(3,5-difluorophenyl)-3-(2'-(4-methylpiperazin-1-yl)-[2,4'-bipyridyl]-6-yl)-1H-indazole 4

[0240]

[0241] Step 1 Preparation of 5-bromo-1-triphenyl-1H-indazole 4b

[0242] Compound 4a (23 g, 117 mmol) and triethylamine (28 g, 280 mmol) were dissolved in dichloromethane (200 ml) and THF (40 mL), and triphenylmethane (39 g, 140 mmol) was added to the solution. The reaction solution was stirred at room temperature overnight. The reaction solution was washed with water and saturated brine, respectively, and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 4b (51 g, 117 mmol), with a yield of 100%.

[0243] MS m / z(ESI):243[M-195] + .

[0244] Step 2 Preparation of (3,5-difluorophenyl)(1-triphenyl-1H-indazol-5-yl)methanol 4c

[0245] Compound 4b (13.2 g, 30 mmol) was dissolved in tetrahydrofuran (100 mL), the solution was cooled to -78 ° C, and n-butyl lithium (14.4 ml, 36 mmol, 2.5 M in hexane) was added dropwise to the solution. The reaction solution was stirred at -78 ° C for 1 h. Then a solution of 3,5-difluorobenzaldehyde (4.7 g, 33 mmol) in tetrahydrofuran (50 ml) was added dropwise to the reaction solution. Then the reaction solution was stirred at -78 ° C for 1 h. The reaction solution was quenched with saturated ammonium chloride solution and extracted with dichloromethane (100 ml*2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain the title compound 4c (6.5 g, 12.9 mmol) with a yield of 43%.

[0246] MS m / z(ESI):282[M-243+Na] + .

[0247] Step 3 Preparation of (3,5-difluorophenyl)-(1H-indazol-5-yl)methanol 4d

[0248] Compound 4c (6 g, 12 mmol) and sodium iodide (9 g, 60 mmol) were added to acetonitrile (150 ml), the solution was cooled to 0°C, trimethylsilyl chloride (6.5 g, 60 mmol) was added to the solution in batches, the reaction solution was stirred at room temperature for 2 hours, then quenched with sodium bisulfite at 0°C, spin-dried, added with water (50 ml), extracted with ethyl acetate (100 ml*2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain the title compound 4d (1.7 g, 33 mmol) with a yield of 55%.

[0249] MS m / z(ESI):261[M+H] + .

[0250] Step 4 Preparation of 5-(3,5-difluorophenyl)-1H-indazole 4e

[0251] Compound 4d (1.7 g, 6.54 mmol) was dissolved in triethylsilane (5 ml) and trifluoroacetic acid (20 ml), and the reaction solution was stirred at 60°C for 3 hours. The reaction solution was then dried by spin-drying and dissolved in ethyl acetate (80 ml), and washed with sodium bicarbonate solution. The organic phase was dried over anhydrous sodium sulfate, filtered and spin-dried to obtain compound 4e (915 mg, 3.75 mmol), with a yield of 57%.

[0252] MS m / z(ESI):245[M+H] + .

[0253] Step 5 Preparation of 3-bromo-5-(3,5-difluorophenyl)-1H-indazole 4f

[0254] Compound 1e (2.0 g, 8.2 mmol) was dissolved in acetonitrile (30 ml), and N-bromosuccinimide (2.9 g, 16.4 mmol) was added to the solution. The reaction solution was stirred at 82 °C for 2 hours. After cooling, the reaction solution was spin-dried to obtain a crude product, which was purified by column chromatography to obtain the title compound 4f (1.8 g, 5.57 mmol) with a yield of 68%.

[0255] MS m / z(ESI):325[M+H] + .

[0256] Step 6 Preparation of 4 g of 3-bromo-5-(3,5-difluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole

[0257] Compound 4f (1.4 g, 4.33 mmol) was dissolved in DMF (10 ml), the solution was cooled to 0°C, sodium hydrogen (208 mg, 5.2 mmol) was added to the solution, the reaction solution was stirred at 0°C for 15 minutes, chloromethyl trisilyl ethyl ether (866 mg, 5.2 mmol) was added, and then stirring was continued at room temperature for 2 hours, saturated ammonium chloride solution was added to the reaction solution, and extracted twice with ethyl acetate (50 ml). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain compound 4g (1.7 g, 3.75 mmol) with a yield of 87%.

[0258] MS m / z(ESI):453[M+H] + .

[0259] Step 7 Preparation of 5-(3,5-difluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-3-(trimethyltinyl)-1H-indazole 4h

[0260] Compound 4g (500 mg, 1.1 mmol), hexamethyltin (1.1 g, 3.3 mmol) and tetrakistriphenylphosphine palladium (127 mg, 0.11 mmol) were suspended in 1,4-dioxane (20 ml), and the reaction solution was stirred at 110 ° C for 18 h. The reaction solution was cooled to room temperature, KF solution (3.3 ml, 1 M) was added, and extracted with ethyl acetate (50 ml). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain compound 4h (480 mg, 0.89 mmol) with a yield of 81%.

[0261] MS m / z(ESI):539[M+H] + .

[0262] Step 8 Preparation of 5-(3,5-difluorophenyl)-3-(2'-(4-methylpiperazin-1-yl)-[2,4'-bipyridyl]-6-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole 4i

[0263] Compound 4h (200 mg, 0.37 mmol), 6-bromo-2'-(4-methylpiperazine-1-yl)-2,4'-bipyridine 4k (124 mg, 0.37 mmol), tetrakistriphenylphosphine palladium (46 mg, 0.04 mmol), cuprous iodide (15 mg, 0.08 mmol) and triethylamine (75 mg, 0.74 mmol) were suspended in DMF (6 ml), and the reaction solution was stirred at 100 ° C for 2 h under nitrogen protection. The reaction solution was cooled to room temperature, water (50 ml) was added, and extracted twice with dichloromethane (30 ml). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain compound 4i (140 mg, 0.22 mmol) with a yield of 60%.

[0264] MS m / z(ESI):627[M+H] + .

[0265] Step 9 Preparation of (5-(3,5-difluorophenyl)-3-(2'-(4-methylpiperazine-1-yl)-[2,4'-bipyridyl]-6-yl)-1H-indazol-1-yl)methanol 4j

[0266] Compound 4i (120 mg, 0.19 mmol) was dissolved in dichloromethane (8 ml), trifluoroacetic acid (4 ml) was added to the solution, and the reaction solution was stirred at room temperature for 2 hours. The reaction solution was spin-dried and the obtained product 4j was directly used for the next reaction.

[0267] MS m / z(ESI):527[M+H] + .

[0268] Step 10 Preparation of 5-(3,5-difluorophenyl)-3-(2'-(4-methylpiperazine-1-yl)-[2,4'-bipyridyl]-6-yl)-1H-indazole 4

[0269] Compound 4j (100 mg, 0.19 mmol) was dissolved in tetrahydrofuran (8 ml), 2N LiOH (4 ml) was added to the solution, and the reaction solution was stirred at room temperature overnight. Water (20 ml) was added, and the mixture was extracted twice with dichloromethane (50 ml). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain compound 4 (20 mg, 0.04 mmol) with a yield of 21%.

[0270] MS m / z(ESI):497[M+H]+ .

[0271] 1H NMR(400MHz,Chloroform-d)δ8.69–8.65(m,1H),8.33(dd,J=5.3,0.8Hz,1H),8.20(dd,J=7.9, 1.0Hz,1H),7.86(t,J=7.8Hz,1H),7.71(dd,J=7.8,0.9Hz,1H),7.57–7.51(m,1H),7.47(dd,J=8 .6,0.8Hz,1H),7.26(dd,J=5.3,1.3Hz,1H),7.20(dd,J=8.6,1.6Hz,1H),6.74–6.66(m,2H),6.6 2(tt,J=9.0,2.4Hz,1H),4.10(s,2H),3.73(t,J=5.0Hz,4H),2.56(t,J=5.0Hz,4H),2.36(s,3H)

[0272] Example 5

[0273] Preparation of 6-(5-(3,5-difluorophenyl)-1H-indazol-3-yl)-N-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)pyridin-2-amine 5

[0274]

[0275] Step 1 Preparation of 6-(5-(3,5-difluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-3-yl)-N-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)pyridin-2-amine 5b

[0276] Compound 4h (200 mg, 0.37 mmol), 6-bromo-nitrogen-(2-methoxy-4-(4-methylpiperazine-1-yl)phenyl)pyridin-2-amine 5a (140 mg, 0.37 mmol), tetrakistriphenylphosphine palladium (46 mg, 0.04 mmol), cuprous iodide (15 mg, 0.08 mmol) and triethylamine (75 mg, 0.74 mmol) were suspended in DMF (6 ml), and the reaction solution was stirred at 100 ° C for 2 h under nitrogen protection. The reaction solution was cooled to room temperature, water (50 ml) was added, and extracted twice with dichloromethane (30 ml). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain compound 5b (146 mg, 0.22 mmol) with a yield of 59%.

[0277] MS m / z(ESI):671[M+H] + .

[0278] Step 2 Preparation of (5-(3,5-difluorophenyl)-3-(6-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyridin-2-yl)-1H-indazol-1-yl)methanol 5c

[0279] Compound 5b (146 mg, 0.22 mmol) was dissolved in dichloromethane (8 ml), trifluoroacetic acid (2 ml) was added to the solution, the reaction solution was stirred at room temperature for 2 hours, the reaction solution was spin-dried, and the obtained product 5c was directly used for the next reaction.

[0280] MS m / z(ESI):571[M+H] + .

[0281] Step 10 Preparation of 6-(5-(3,5-difluorophenyl)-1H-indazol-3-yl)-N-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)pyridin-2-amine 5

[0282] Compound 5c (100 mg, 0.18 mmol) was dissolved in tetrahydrofuran (10 ml), 2N LiOH (10 ml) was added to the solution, and the reaction solution was stirred at room temperature for 4 hours. Water (20 ml) was added, and the mixture was extracted twice with dichloromethane (50 ml). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain compound 5 (40 mg, 0.074 mmol) with a yield of 42%.

[0283] MS m / z(ESI):541[M+H] + .

[0284] 1H NMR(400MHz,DMSO-d6)δ13.18(s,1H),8.26(s,1H),7.67–7.56(m,1H),7.4 9(d,J=8.5Hz,1H),7.42(d,J=7.4Hz,1H),7.37–7.29(m,1H),7.01(d,J=8.3 Hz,3H),6.82(d,J=8.5Hz,1H),6.65–6.55(m,1H),6.49–6.31(m,2H),4.53( s,1H),4.11(s,2H),3.77(s,3H),3.65(br,4H),3.07(br,4H),2.64(s,3H).

[0285] Example 6

[0286] Preparation of N-(5-(1-(3,5-difluorophenyl)cyclopropyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzoamide 6

[0287]

[0288]

[0289] The first step is the preparation of 1-(3-bromo-4-fluorophenyl)-1-(3,5-difluorophenyl)ethyl-1-ol 6c

[0290] Compound 6a (10 g, 51.8 mmol) was dissolved in tetrahydrofuran (150 mL), the solution was cooled to -78 ° C, n-butyl lithium solution (22.8 mL, 57 mmol) was added dropwise to the solution, and stirred at -78 ° C for 1 hour, and then compound 6b (11.2 g, 51.8 mmol) in tetrahydrofuran (50 mL) was added, and the reaction solution was stirred at room temperature overnight. The reaction solution was quenched with saturated ammonium chloride solution and extracted with ethyl acetate (200 mL). The organic phase was dried and then spin-dried to obtain a crude product of compound 6c (19 g).

[0291] MS m / z(ESI):313[M-17] + .

[0292] Step 2 Preparation of 2-bromo-4-(1-(3,5-difluorophenyl)vinyl)-1-fluorobenzene 6d

[0293] Compound 6c (19 g, 57.5 mmol) was dissolved in acetic acid (30 mL), and concentrated sulfuric acid (3 mL) was added dropwise under ice bath. The reaction solution was stirred at room temperature for 30 minutes. The reaction solution was added into ice water (200 mL), extracted with ethyl acetate (200 mL), the organic phase was washed with saturated sodium bicarbonate solution and brine, dried with anhydrous sodium sulfate, filtered and dried to obtain the initial product, and purified by silica gel column chromatography to obtain compound 6d (11.7 g, 37.4 mmol), with a yield of 65%.

[0294] Step 3 Preparation of 2-bromo-4-(1-(3,5-difluorophenyl)cyclopropyl)-1-fluorobenzene 6e

[0295] NaH (392 mg, 9.8 mmol) was dissolved in DMSO (10 mL), cooled to 0°C, (CH3)3SOI (5.87 g, 26.7 mmol) was added, and the reaction solution was stirred at room temperature for 2 hours. A solution of 6d (2.8 g, 8.9 mmol) in dimethyl sulfoxide (10 mL) was added to the reaction solution, and the reaction solution was stirred at room temperature for 1 hour. The reaction solution was quenched with saturated ammonium chloride solution, extracted with ethyl acetate (100 mL), and the organic phase was washed with saturated brine, dried and spin-dried to give a crude product, which was purified by column chromatography to give compound 6e (1.8 g, 5.5 mmol) with a yield of 62%.

[0296] Step 4 Preparation of 5-(1-(3,5-difluorophenyl)cyclopropyl)-2-fluorobenzonitrile 6f

[0297] Compound 6e (0.8 g, 2.45 mmol), zinc cyanide (286 mg, 2.45 mmol) and tetrakistriphenylphosphine palladium (288 mg, 0.25 mmol) were mixed in DMF (6 mL), replaced with nitrogen twice, and the reaction solution was reacted at 110 ° C in a microwave for 1 hour. After cooling, the reaction solution was poured into water (50 mL) and extracted with ethyl acetate (50 mL). The organic phase was dried and spin-dried to obtain a crude product, which was purified by column chromatography (PE / EA=30:1) to obtain a crude product of compound 6f (760 mg).

[0298] MS m / z(ESI):274[M+H] + .

[0299] Step 5 Preparation of 5-(1-(3,5-difluorophenyl)cyclopropyl)-1H-indazole-3-amine 6g

[0300] Compound 6f (680 mg, 2.49 mmol) and hydrazine hydrate (1.25 g, 24.9 mmol) were mixed in N-methylpyrrolidone (6 mL). The reaction solution was reacted at 120 ° C in a microwave reactor for 1 hour. After cooling, it was added to (30 mL) and extracted with ethyl acetate (50 mL). The organic phase was dried and spin-dried to obtain a crude product. After purification by column chromatography (PE / EA=30:1), compound 6g (500 mg, 1.75 mmol) was obtained with a yield of 71%.

[0301] MS m / z(ESI):286[M+H] + .

[0302] Step 6 Preparation of N-(5-(1-(3,5-difluorophenyl)cyclopropyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetyl)benzamide 6h

[0303] A solution of compound 6g (285 mg, 1.0 mmol) and DIPEA (387 mg, 3.0 mmol) in tetrahydrofuran (15 mL) was added dropwise to the tetrahydrofuran (5 mL) coolant of the above 1f, and the reaction was stirred at -20°C for 2 hours, and then continued to stir at room temperature for 2 hours. The reaction solution was diluted with dichloromethane (50 mL), washed with mixed brine, dried and spin-dried to obtain a crude product, which was purified by column chromatography (DCM / MeOH=10:1) to obtain compound 6h (300 mg, 0.44 mmol) with a yield of 44%.

[0304] MS m / z(ESI):683[M+H] + .

[0305] Step 7: N-(5-(1-(3,5-difluorophenyl)cyclopropyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 6

[0306] Compound 6h (300 mg, 0.44 mmol) was dissolved in a mixed solvent of methanol (10 mL) and water (5 mL), potassium carbonate (182 mg, 1.32 mmol) was added, the reaction solution was stirred at room temperature for 3 hours, water (20 ml) was added, and it was extracted with dichloromethane (50 ml). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain compound 6 (40 mg, 0.075 mmol), with a yield of 16%.

[0307] MS m / z(ESI):587[M+H] + .

[0308] 1 H NMR (400MHz, DMSO-d6) δ12.58(s,1H),10.05(s,1H),8.34(d,J=7.7Hz,1H),7.76(d,J=8.9H z,1H),7.49(s,1H),7.40–7.21(m,3H),7.01(t,J=8.0Hz,2H),6.21(dd,J=9.2,2.2Hz,1H), 6.11(d,J=2.4Hz,1H),3.87–3.74(m,2H),3.75–3.65(m,1H),3.50(t,J=10.6Hz,2H),3.23( d,J=5.1Hz,4H),2.41(t,J=4.8Hz,4H),2.20(s,3H),1.97–1.86(m,2H),1.36-1.14(m,6H).

[0309] Example 7

[0310] Preparation of N-(5-((3,5-difluorophenyl)difluoromethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 7

[0311]

[0312] Step 1 Preparation of 5-(3,5-difluorobenzoyl)-2-fluorobenzonitrile 7a

[0313] Compound 5-((3,5-difluorophenyl)(hydroxy)methyl)-2-fluorobenzonitrile I-2c (2 g, 7.6 mmol) was dissolved in DCM (20 mL), Dess Martin (6.4 g, 15.2 mmol) was added under ice bath, and the temperature was slowly raised to room temperature under stirring. The mixture was stirred at room temperature for 1 hour. The reaction solution was filtered, the filtrate was neutralized with saturated NaHCO3 aqueous solution, the mixture was extracted with EA, the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography (PE:EA=4:1) to obtain 5-(3,5-difluorobenzoyl)-2-fluorobenzonitrile 7a (1.1 g, 4.2 mmol), with a yield of 55%.

[0314] MS m / z(ESI):262[M+H] + .

[0315] Step 2 Preparation of 5-((3,5-difluorophenyl)difluoromethyl)-2-fluorobenzonitrile 7b

[0316] Compound 5-(3,5-difluorobenzoyl)-2-fluorobenzonitrile 7a (130 mg, 0.5 mmol) and BAST (1 mL) were mixed together, and the mixture was stirred at 110° C. for 3 hours. The reaction solution was cooled, quenched with water, extracted three times with DCM, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography (PE:EA=4:1) to obtain 5-((3,5-difluorophenyl)difluoromethyl)-2-fluorobenzonitrile 7b (100 mg, 0.35 mmol) as a colorless oil in a yield of 70%.

[0317] MS m / z(ESI):284[M+H] + .

[0318] Step 3 Preparation of 5-((3,5-difluorophenyl)difluoromethyl)-1H-indazol-3-amine 7c

[0319] 5-((3,5-difluorophenyl)difluoromethyl)-2-fluorobenzonitrile 7b (100 mg, 0.35 mmol) and hydrazine hydrate (100 mg, 3.5 mmol) were dispersed in THF (5 mL), and the mixture was stirred at 45 ° C for 2 hours. After the reaction solution was cooled, it was quenched with concentrated hydrochloric acid, THF was evaporated, water was added to dilute it, and then it was neutralized to neutrality with ammonia water. A yellow solid precipitated, which was filtered and dried to obtain a yellow solid 5-((3,5-difluorophenyl)difluoromethyl)-1H-indazole-3-amine 7c (100 mg, 0.33 mmol) as a light yellow solid with a yield of 96%.

[0320] MS m / z(ESI):296[M+H] + .

[0321] Step 4 Preparation of N-(5-((3,5-difluorophenyl)difluoromethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 7d

[0322] A THF solution of 4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetylamino)benzoyl chloride 1f (104 mg, 0.24 mmol) was added dropwise to a THF solution of 5-((3,5-difluorophenyl)difluoromethyl)-1H-indazol-3-amine 7c (80 mg, 0.24 mmol) and triethylamine (73 mg, 0.72 mmol) at -20°C. After the addition was complete, the reaction mixture was stirred at -20°C. The reaction mixture was stirred for 1 hour, and the reaction solution was warmed to room temperature, quenched with water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography to give N-(5-((3,5-difluorophenyl)difluoromethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 7d (60 mg, 0.08 mmol) as a yellow solid in a yield of 36%.

[0323] MS m / z(ESI):693[M+H] + .

[0324] Step 6 Preparation of N-(5-((3,5-difluorophenyl)difluoromethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 7

[0325] N-(5-((3,5-difluorophenyl)difluoromethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 7d (60 mg, 0.08 mmol) was dissolved in methanol (2 mL), and triethylamine (2 mL, was added. The reaction mixture was stirred at room temperature for 6 hours and concentrated. The crude product was purified by column chromatography (DCM:MeOH=10:1) to give N-(5-((3,5-difluorophenyl)difluoromethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 7 (5 mg, 0.008 mmol) as a light yellow solid in a yield of 10%.

[0326] MS m / z(ESI):597[M+H]+

[0327] 1 H NMR (400MHz, DMSO-d6) δ10.05(s,1H),8.67(s,1H),8.09–8.01(m,1H),7.71(s,1H),7.64–7. 52(m,2H),7.45–7.37(m,1H),7.35–7.29(m,2H),6.88(d,J=9.0Hz,1H),6.21–6.13(m,1H),6 .06(d,J=2.2Hz,1H),3.80–3.72(m,2H),3.65-3.57(m,1H),3.43(t,J=10.5Hz,2H),3.21(t, J=5.1Hz,4H),2.38(t,J=5.1Hz,4H),2.18(s,3H),1.86(d,J=12.9Hz,2H),1.32–1.23(m,2H).

[0328] Example 8

[0329] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((4-methylpiperazin-1-yl)sulfonyl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 8

[0330]

[0331]

[0332] Step 1 Preparation of ethyl 4-fluoro-2-nitrobenzoate 8b

[0333] Add 8a (9.2 g, 50 mmol), potassium carbonate (1.38 g, 100 mmol) and DMF (50 mL) to a dry reaction bottle, then drop iodoethane (9.4 g, 60 mmol). Stir overnight at room temperature after addition. TLC (petroleum ether: ethyl acetate = 2:1) shows that the reaction is complete. Add water and ethyl acetate to separate the liquids, wash the ethyl acetate layer with water several times, dry over anhydrous sodium sulfate, and evaporate to dryness under reduced pressure to obtain compound 8b (9.6 g, 45 mmol, 90%).

[0334] MS m / z(ESI):214[M+H] +

[0335] Step 2 Preparation of ethyl 4-((3-((2-ethylhexyl)oxy)-3-carbonylpropyl)thio)-2-nitrobenzoate 8d

[0336] Add compound 8b (2.14 g, 10 mmol), compound 8c (2.6 g, 12 mmol), DMF (10 mL) and triethylamine (2.0 g, 20 mmol) to the reaction flask. The mixture was reacted at 90-100°C for 4 hours. TLC (petroleum ether: ethyl acetate = 2:1) showed that the raw materials had reacted completely. Pour into water, extract with ethyl acetate, wash with water several times, dry over anhydrous sodium sulfate and purify with flash (petroleum ether: ethyl acetate = 5:1) to obtain compound 8d (3.0 g, 7.5 mmol, 75%).

[0337] MS m / z(ESI):434[M+Na] +

[0338] Step 3 Preparation of ethyl 4-mercapto-2-nitrobenzoate 8e

[0339] Compound 8d (0.41 g, 1.0 mmol) and ethanol (10 ml) were added to the reaction flask, and sodium ethoxide (0.200 g, 3 mmol) was added after the solution was clarified. The mixture was stirred at room temperature for 4 hours. TLC (petroleum ether: ethyl acetate = 2:1) showed that the reaction of the raw materials was complete. Citric acid was added for neutralization, and ethyl acetate was extracted 3 times. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and dried under reduced pressure to obtain 0.4 g of crude compound 8e, which was directly used in the next step.

[0340] Step 4 Preparation of Compound Ethyl 4-(Chlorosulfonyl)-2-nitrobenzoate 8f

[0341] Mix 2M HCl and 5 times the volume of acetonitrile solution, add NCS (0.79 g, 6 mmol), and ice bath to about 10°C. After the solution is clarified, add acetonitrile solution of compound 8e (0.4 g crude product) and stir at about 10°C for 2 hours. TLC (petroleum ether: ethyl acetate = 2:1) shows that the raw material has reacted. Add ether to dilute the reaction solution, and add water to split. Wash the organic phase with a small amount of dilute sodium bicarbonate aqueous solution, dry over anhydrous sodium sulfate, and evaporate to dryness under reduced pressure at low temperature to obtain a crude compound 8f (0.4 g).

[0342] Step 5 Preparation of 8 g of ethyl 4-((4-methylpiperazine-1-yl)sulfonyl)-2-nitrobenzoate

[0343] Compound 8f (0.4 g) was dissolved in dichloromethane, 1.0 mL of methylpiperazine was added, and the mixture was stirred at room temperature for 1 hour. TLC (dichloromethane: methanol = 10:1) showed that the reaction was complete. Silica gel was added and the sample was mixed. Flash purification (dichloromethane: methanol = 20:1) gave compound 8g (0.22 g, 0.8 mmol, 61%).

[0344] MS m / z(ESI):358[M+H]+

[0345] Step 6 Preparation of ethyl 2-amino-4-((4-methylpiperazin-1-yl)sulfonyl)benzoate 8h

[0346] Compound 8g (0.36 g, 1.0 mmol), reduced iron powder (0.28 g, 5 mmol), THF (10 ml), AcOH (0.5 mL), and water (0.5 ml) were added to the reaction flask. The reaction mixture was refluxed at 60 ° C for 2 hours. TLC (dichloromethane: methanol = 10: 1) showed that the reaction was complete. A large amount of ethyl acetate was added to dilute and filtered. After the filtrate was concentrated, ethyl acetate was added, and the mixture was washed with sodium bicarbonate aqueous solution. After separation, the organic phase was dried over anhydrous sodium sulfate. After reduced pressure concentration, compound 8h (0.26 g, 0.8 mmol, 80%) was obtained.

[0347] MS m / z(ESI):328[M+H] +

[0348] Step 7 Preparation of ethyl 4-((4-methylpiperazin-1-yl)sulfonyl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzoate 8i

[0349] Compound 8h (0.33 g, 1.0 mmol), TFA (1.0 ml) and NaBH(OAc)3 (0.88 g, 4.0 mmol) were added to a dry reaction bottle. After stirring at room temperature for 10 min, a dichloromethane solution of compound 4-dihydropyrone (0.5 g, 5 mmol) was slowly added. After 10 min, TLC (dichloromethane: methanol = 10:1) showed that the reaction was complete. The reaction solution was poured into a sodium bicarbonate aqueous solution, stirred for half an hour, and then separated. The organic phase was separated and washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. It was flash purified (dichloromethane: methanol = 10:1) to obtain compound 8i (0.28 g, 0.7 mmol, 70%).

[0350] MS m / z(ESI):412[M+H] +

[0351] Step 8 Preparation of 4-((4-methylpiperazin-1-yl)sulfonyl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzoic acid 8j

[0352] Compound 8i (4.11 g, 10.0 mmol) and THF (50 ml) were added to the reaction flask. After the solution was clarified, LiOH (4 M, 10 ml) and a small amount of water were added. After stirring overnight, TLC (dichloromethane: methanol = 20:1) showed that the reaction was complete. Citric acid was added for neutralization, and ethyl acetate was extracted twice. The organic phases were combined and washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 8j (3.07 g, 8.0 mmol, 80%).

[0353] MS m / z(ESI):384[M+H] +

[0354] Step 9 Preparation of 4-((4-methylpiperazine-1-yl)sulfonyl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoic acid 8k

[0355] Compound 8j (0.38 g, 1.0 mmol) was suspended in dichloromethane, TFAA was added until the solution was clear, and then the reaction solution was concentrated under reduced pressure. The obtained solid was dried with petroleum ether, filtered, added with water, stirred for 48 hours, filtered, and dried. The solid was removed under high temperature and reduced pressure to obtain compound 8k (0.33 g, 0.7 mmol, 70%).

[0356] Step 10 Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((4-methylpiperazin-1-yl)sulfonyl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 81

[0357] Under nitrogen protection, compound 8k (48 mg, 0.1 mmol), dichloromethane and a drop of DMF were added to the dried reaction bottle, and then oxalyl chloride (0.3 ml) was added. After stirring at room temperature for 4 hours, LCMS showed that the raw material was completely converted. The reaction solution was concentrated under reduced pressure and dried with dichloromethane. The resulting solid was dissolved in THF and cooled to -40°C (external temperature) under nitrogen protection. Then, a THF solution of compound I-2 (26 mg, 0.1 mmol) and DIPEA (0.1 ml) was slowly added, and the external temperature was kept at -40 to -30°C. After the addition, the reaction solution was allowed to heat up naturally, and LCMS showed that the main peak was the product point. The reaction solution was concentrated to a smaller volume and purified by PLC (dichloromethane: methanol = 10:1) to obtain compound 8l (30 mg, 0.04 mmol).

[0358] MS m / z(ESI):721[M+H] +

[0359] Step 11 Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((4-methylpiperazin-1-yl)sulfonyl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 8

[0360] Compound 81 (30 mg, 0.04 mmol) and methanol were added to the reaction flask. After the solution was clarified, a small amount of potassium carbonate aqueous solution was added. TLC (dichloromethane: methanol = 10: 1) showed that the reaction was complete. Ethyl acetate was added for dilution, and the organic phase was washed with a small amount of brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by PLC (dichloromethane: methanol = 10: 1) to obtain the target product 8 (20 mg, 0.03 mmol, 76%).

[0361] MS m / z(ESI):625[M+H] +

[0362] 1 H NMR (400MHz, DMSO-d6) δ12.76(s,1H),10.75(s,1H),8.06(d,J=8.2Hz,1H),7.94(d,J=7.9Hz ,1H),7.53(s,1H),7.41(d,J=8.6Hz,1H),7.25(dd,J=8.7,1.6Hz,1H),7.02–6.93(m,4H),6.8 9(dd,J=8.2,1.7Hz,1H),4.02(s,2H),3.80(dt,J=11.8,3.9Hz,2H),3.75-3.65(m,1H),3.55- 3.42(m,2H),2.95(br,4H),2.35(br,4H),2.14(s,3H),1.94-1.85(m,2H),1.45-1.34(m,2H).

[0363] Example 9

[0364] Preparation of N-(5-(3,5-difluorobenzyl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 9

[0365]

[0366] Step 1 Preparation of 4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 9a

[0367] 4-(4-Methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoyl chloride 1f (0.86 g, 2 mmol) was added to a mixed solvent of aqueous ammonia (5 mL) and dichloromethane, stirred overnight, the dichloromethane layer was separated, concentrated, and purified by column chromatography (DCM:MeOH=15:1) to obtain the product 9a (0.25 g, 0.8 mmol), with a yield of 39%.

[0368] MS m / z(ESI):319[M+H] +

[0369] Step 2 Preparation of 5-(3,5-difluorobenzyl)pyrazolo[1,5-a]pyrimidine 9d

[0370] Zinc powder (3 g, 46 mmol) was dispersed in THF (100 mL), and 0.2 mL of 1,2-dichloroethane was added. The mixture was stirred at 50 m°C for 1 hour under nitrogen protection. After cooling, 1-(bromomethyl)-3,5-difluorobenzene (7.2 g, 35 mmol) was added at 0°C, and then 0.2 mL of TMSCl was added. The reaction mixture was stirred at room temperature for 6 hours to obtain 9c. The reaction solution was slowly dripped into a THF solution of 5-chloropyrazolo[1,5-a]pyrimidine 9b (1.5 g, 9.8 mmol) at room temperature, and Pd(pph3)4 (1.1 g, 0.98 mmol) was added. The reaction mixture was stirred at 60°C for 1 hour under nitrogen protection, and saturated aqueous ammonium chloride was added to quench after cooling. The mixture was extracted with ethyl acetate three times, and the organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (PE:EA=5:1) to obtain 5-(3,5-difluorobenzyl)pyrazolo[1,5-a]pyrimidine 9d (2.2 g, 8.9 mmol) as a yellow solid in a yield of 90%.

[0371] MS m / z(ESI):246[M+H] +

[0372] Step 3 Preparation of 5-(3,5-difluorobenzyl)-3-iodopyrazolo[1,5-a]pyrimidine 9e

[0373] NIS (918 mg, 4 mmol) was added to 5-(3,5-difluorobenzyl)-3-iodopyrazolo[1,5-a]pyrimidine 9d (500 mg, 2 mmol) in THF, and the reaction mixture was reacted at RT for 2 hours. The mixture was quenched with sodium thiosulfate aqueous solution, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography to give 5-(3,5-difluorobenzyl)-3-iodopyrazolo[1,5-a]pyrimidine 9e (600 mg, 1.61 mmol) as a yellow solid in a yield of 80%.

[0374] MS m / z(ESI):372[M+H] +

[0375] Step 4 Preparation of N-(5-(3,5-difluorobenzyl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 9

[0376] A mixture of 5-(3,5-difluorobenzyl)-3-iodopyrazolo[1,5-a]pyrimidine 9e (100 mg, 0.27 mmol), 4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 9a (85 mg, 0.27 mmol), CuI (25 mg, 0.13 mmol), copper powder (8 mg, 0.13 mmol), K3PO4 (114 mg, 0.54 mmol) and N1,N2-dimethylethane-1,2-diamine (11 mg, 0.13 mmol) was dispersed in 4 mL of toluene, and the mixture was stirred at 110°C for 56 hours. The reaction solution was cooled and filtered. The filter cake was washed three times with dichloromethane. The filtrates were combined and concentrated. The crude product was purified by column chromatography (DCM:MeOH=10:1) to give N-(5-(3,5-difluorobenzyl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 9 (9 mg, 0.016 mmol) as a light yellow solid with a yield of 6%.

[0377] MS m / z(ESI):562[M+H] +

[0378] 1H NMR(400MHz,DMSO-d6)δ9.56(s,1H),8.91(d,J=7.2Hz,1H),8.27(s,1H),8.22(d,J=7.7Hz,1H), 7.74(d,J=9.0Hz,1H),7.12–7.04(m,3H),6.90(d,J=7.2Hz,1H),6.23(dd,J=8.9,2.2Hz,1H),6. 10(d,J=2.3Hz,1H),4.14(s,2H),3.86–3.71(m,2H),3.68–3.57(m,1H),3.49–3.40(m,2H),3.22 (t,J=5.0Hz,4H),2.40(t,J=5.1Hz,4H),2.19(s,3H),1.90(d,J=12.7Hz,2H),1.38–1.28(m,2H).

[0379] Example 10

[0380] Preparation of N-(5-(1-(3,5-difluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 10

[0381]

[0382]

[0383] Step 1 Preparation of (2-(3-bromo-4-fluorophenyl)-2-(3,5-difluorophenyl)ethyl)boronic acid 10a

[0384] A solution of compound 6d (8.9 g, 28.4 mmol) in tetrahydrofuran (150 mL) was added dropwise to a borane tetrahydrofuran complex (56.8 mL, 56.8 mmol) at 0°C. The reaction solution was stirred at room temperature for 2 hours, and then ice water (150 mL) was carefully added to the reaction solution. The reaction solution was stirred at room temperature for 3 hours. The reaction solution was concentrated and extracted with ethyl acetate (200 mL). The organic phase was washed with a saturated sodium bicarbonate solution and brine, respectively, and dried and then spin-dried to obtain a crude product. After column chromatography purification, a crude product of compound 10a (10 g) was obtained. MS m / z (ESI): 383 [M+23] + .

[0385] Step 2 Preparation of 2-(3-bromo-4-fluorophenyl)-2-(3,5-difluorophenyl)ethyl-1-ol 10b

[0386] Compound 10a (10 g, 27.8 mmol) was dissolved in tetrahydrofuran (60 mL), and 3M sodium hydroxide aqueous solution (60 mL) was added. The reaction solution was cooled to 0°C, and 30% hydrogen peroxide (30 mL) was slowly added dropwise. The reaction solution was stirred at room temperature for 3 hours, and then quenched with saturated sodium thiosulfate solution. It was extracted with ethyl acetate (200 mL), and the organic phase was washed with saturated sodium bicarbonate solution and brine, dried with anhydrous sodium sulfate, filtered and dried to obtain the initial product, and purified by silica gel column chromatography to obtain compound 10b (4 g, 12 mmol), with a yield of 43%.

[0387] MS m / z(ESI):315[M-17] + .

[0388] Step 3 Preparation of 4-(2-(benzyloxy)-1-(3,5-difluorophenyl)ethyl)-2-bromo-1-fluorobenzene 10c

[0389] Compound 10b (1 g, 3 mmol) was dissolved in tetrahydrofuran (15 mL), cooled to 0°C, sodium hydroxide (144 mg, 3.6 mmol) was added, and the reaction solution was stirred at 0°C for 1 hour. Benzyl bromide (615.6 mg, 3.6 mmol) was added to the reaction solution, and the reaction solution was stirred at 70°C for 5 hours. After cooling, the reaction solution was poured into water (50 mL), extracted with ethyl acetate (50 mL), and the organic phase was washed with saturated brine, dried and spin-dried to obtain a crude product, which was purified by column chromatography to obtain compound 10c (320 mg, 0.76 mmol) with a yield of 25%.

[0390] Step 4 Preparation of 5-(2-(benzyloxy)-1-(3,5-difluorophenyl)ethyl)-2-fluorobenzonitrile 10d

[0391] Compound 10c (160 mg, 0.38 mmol), zinc cyanide (45 mg, 0.38 mmol) and tetrakistriphenylphosphine palladium (46 mg, 0.04 mmol) were mixed in DMF (3 mL), replaced with nitrogen twice, and the reaction solution was reacted at 110 ° C. on a microwave for 1 hour. After cooling, the reaction solution was poured into water (30 mL) and extracted with ethyl acetate (30 mL). The organic phase was dried and spin-dried to obtain a crude product, which was purified by column chromatography (PE / EA=30:1) to obtain compound 10d (113 mg, 0.31 mmol) with a yield of 81%.

[0392] MS m / z(ESI):368[M+H] + .

[0393] Step 5 Preparation of 5-(2-(benzyloxy)-1-(3,5-difluorophenyl)ethyl)-1H-indazol-3-amine 10e

[0394] Compound 10d (230 mg, 0.63 mmol) and hydrazine hydrate (313 mg, 6.3 mmol) were mixed in N-methylpyrrolidone (6 mL), and the reaction solution was reacted at 120 ° C in a microwave reactor for 1 hour. After cooling, it was added to (30 mL) and extracted with ethyl acetate (50 mL). The organic phase was dried and spin-dried to obtain a crude product, which was purified by silica gel column chromatography to obtain a crude product of compound 10e (400 mg).

[0395] MS m / z(ESI):380[M+H] + .

[0396] Step 6 Preparation of N-(5-(2-(benzyloxy)-1-(3,5-difluorophenyl)ethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 10f

[0397] A solution of compound 10e (400 mg, 1.05 mmol) and DIPEA (387 mg, 3.0 mmol) in tetrahydrofuran (15 mL) was added dropwise to the cooling liquid of tetrahydrofuran (5 mL) of 1f, and the reaction was stirred at -20°C for 2 hours, and then continued to stir at room temperature for 2 hours. The reaction solution was diluted with dichloromethane (50 mL), washed with mixed brine, dried and spin-dried to obtain a crude product, which was purified by column chromatography (DCM / MeOH=10:1) to obtain compound 10f (370 mg, 0.48 mmol) with a yield of 45%.

[0398] MS m / z(ESI):777[M+H] + .

[0399] Step 7 Preparation of 10 g of N-(5-(2-(benzyloxy)-1-(3,5-difluorophenyl)ethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide

[0400] Compound 10f (170 mg, 0.22 mmol) was dissolved in a mixed solvent of methanol (10 mL) and water (5 mL), potassium carbonate (90.7 mg, 0.66 mmol) was added, the reaction solution was stirred at room temperature for 3 hours, water (20 ml) was added, and it was extracted with dichloromethane (50 ml). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 10g (82 mg, 0.12 mmol), with a yield of 55%.

[0401] MS m / z(ESI):681[M+H]+.

[0402] Step 8 Preparation of N-(5-(1-(3,5-difluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 10

[0403] Compound 10g (102mg, 0.15mmol) was dissolved in ethanol (10mL), 10% palladium carbon (20mg) and ammonium formate (190mg, 3mmol) were added, and the reaction solution was stirred at 90°C for 24 hours, cooled and filtered. The filtrate was concentrated to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 10 (12mg, 0.02mmol), with a yield of 16%.

[0404] MS m / z(ESI):591[M+H] + .

[0405] 1 H NMR (400MHz, DMSO-d6) δ12.59(s,1H),10.06(s,1H),8.30(d,J=7.7Hz,1H),7.75(d,J=9.1Hz,1H),7.47(s,1H),7.39 –7.34(m,1H),7.31–7.20(m,2H),7.00(t,J=8.8Hz,2H),6.20(dd,J=9.0,2.3Hz,1H),6.10(d,J=2.3Hz,1H),4.92(t,J =5.4Hz,1H),4.51(t,J=7.8Hz,1H),4.06(t,J=6.6Hz,2H),3.85–3.74(m,2H),3.72–3.64(m,1H),3.48(td,J=11.7,10 .9,2.5Hz,2H),3.23(t,J=5.1Hz,4H),2.40(t,J=5.0Hz,4H),2.19(s,3H),1.91(d,J=13.2Hz,2H),1.41-1.25(m,2H).

[0406] Embodiment 11

[0407] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(oxadiazine-3-ylamino)benzamide 11

[0408]

[0409] The first step is the preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-fluoro-2-nitrobenzamide 11b

[0410] Compound 4-fluoro-2-nitrobenzoic acid I-1a (3.7 g, 20 mmol) was suspended in DCM (60 mL), and oxalyl chloride (6.4 g, 50 mmol) was added under ice-water bath cooling, and stirred for 3 hours. 11a concentrated under reduced pressure was dissolved in tetrahydrofuran (40 mL), and DIEA (5.16 g, 40 mmol) was added. Compound I-2 (3.9 g, 15 mmol) was added under ice-water bath cooling, and stirred overnight. Ethyl acetate and water were added for extraction, and ethyl acetate layer was used for column chromatography to obtain product 11b (1.2 g, 2.8 mmol) with a yield of 14%.

[0411] MS m / z(ESI):427[M+H] + .

[0412] Step 2 Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-nitrobenzamide 11c

[0413] Compound N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-fluoro-2-nitrobenzamide 11b (200 mg, 0.47 mmol), 1-methylpiperazine (93 mg, 0.93 mmol) and DIPEA (181 mg, 1.41 mmol) were dissolved in DMSO (4 mL). The mixture was stirred at 100°C for 3 hours, cooled and quenched with water, extracted with ethyl acetate, the organic phases were combined, washed twice with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (DCM: MeOH = 10: 1) to obtain 200 mg of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-nitrobenzamide 11c as a yellow solid, with a yield of 84%.

[0414] MS m / z(ESI):507[M+H] +

[0415] Step 2 Preparation of 2-amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)benzamide 11d

[0416] Compound N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-nitrobenzamide 11c (200 mg, 0.39 mmol) was dissolved in methanol, Pd / C (10%) was added, and the mixture was stirred at room temperature overnight under hydrogen conditions. The reaction solution was filtered, and the filtrate was concentrated. 150 mg of crude 2-amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)benzamide 11d was directly used in the next step. The yield was 79%.

[0417] MS m / z(ESI):477[M+H] + .

[0418] Step 3 Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(oxadiazine-3-ylamino)benzamide 11

[0419] 2-amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)benzamide 11d (150 mg, 0.31 mmol) and oxetanone (67 mg, 0.93 mmol) were dispersed in DCM (5 mL), trifluoroacetic acid (0.3 mL) was added, and then tetramethylammonium triacetoxyborohydride (414 mg, 1.57 mmol) was added. The mixture was stirred at room temperature for 12 hours, quenched with saturated aqueous NaHCO3 solution, extracted with DCM, and the organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography to obtain 2.5 mg of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(oxetan-3-ylamino)benzamide 11, with a yield of 1.5%.

[0420] MS m / z(ESI):533[M+H] + .

[0421] 1H NMR(400MHz, Methanol-d4)δ7.78(d,J=8.9Hz,1H),7.60(s,1H),7.42(d,J=8.6Hz ,1H),7.23(dd,J=8.6,1.6Hz,1H),6.84–6.78(m,2H),6.74–6.67(m,1H),6.39(dd ,J=8.9,2.4Hz,1H),5.83(d,J=2.4Hz,1H),5.01(t,J=6.6Hz,2H),4.74(p,J=6.3H z,1H),4.53(t,J=6.1Hz,2H),4.06(s,2H),3.46(s,4H),3.00(s,4H),2.65(s,3H).

[0422] Example 12

[0423] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 12

[0424]

[0425] Step 1 Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-nitrobenzamide 12b

[0426] Compound N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-fluoro-2-nitrobenzamide 11b (200 mg, 0.47 mmol), 2-methyloctahydropyrrolo[3,4-c]pyrrole 12a (117 mg, 0.93 mmol) and DIPEA (181 mg, 1.41 mmol) were dissolved in DMSO (4 mL). The mixture was stirred at 150° C. for 3 hours, cooled, quenched with water, extracted with ethyl acetate, the organic phases were combined, washed twice with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (DCM:MeOH=10:1) to obtain 200 mg of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-nitrobenzamide 12b amine as a yellow solid. MS m / z(ESI):533[M+H] + Yield: 84%

[0427] Step 2 Preparation of 2-amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)benzamide 12c

[0428] The compound N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-nitrobenzamide 12b (200 mg, 0.37 mmol) was dissolved in methanol, Pd / C (10%) was added, and the mixture was stirred at room temperature overnight under hydrogen conditions. The reaction solution was filtered, and the filtrate was concentrated. 150 mg of crude 2-amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)benzamide 12c was directly used in the next step. The yield was 80%.

[0429] MS m / z(ESI):503[M+H] + .

[0430] Step 3 Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 12

[0431] 2-Amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)benzamide 12c (150 mg, 0.29 mmol) and tetrahydro-4H-pyran-4-one (87 mg, 0.87 mmol) were dispersed in DCM (5 mL), trifluoroacetic acid (0.3 mL) was added, followed by tetramethylammonium tetraacetoxyborohydride (414 mg, 1.57 mmol). The mixture was stirred at room temperature for 12 hours, quenched with saturated aqueous NaHCO3, extracted with DCM, the organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography to give 1.5 mg of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 12, yield 1.5%.

[0432] MS m / z(ESI):533[M+H]+.

[0433] 1H NMR (400MHz, Methanol-d4) δ7.70(d,J=8.9Hz,1H),7.57(s,1H),7.40(d,J=8.7Hz,1H),7.22(dd,J=8.7,1. 5Hz,1H),6.81(d,J=7.0Hz,2H),6.75–6.66(m,1H),6.08(dd,J=8.8,2.2Hz,1H),5.92(d,J=2.1Hz,1H),4.05 (s,2H),3.91(dt,J=11.6,4.0Hz,2H),3.65–3.61(m,1H),3.59–3.52(m,2H),3.48–3.34(m,4H),3.10–3.06 (m,2H),2.99–2.95(m,2H),2.62(d,J=10.3Hz,2H),2.43(s,3H),2.03(d,J=13.1Hz,2H),1.56–1.48(m,2H).

[0434] Example 13

[0435] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 13

[0436]

[0437] Using the same preparation method as Example 12 and using 2-(pyrrolidin-1-yl)ethylamine as raw material, Compound 13 can be obtained.

[0438] MS m / z(ESI):589[M+H] + .

[0439] 1 H NMR(400MHz, Methanol-d4)δ7.75(d,J=8.9Hz,1H),7.55(s,1H),7.41(d,J=8.6Hz,1H), 7.27–7.22(m,1H),6.81(d,J=6.9Hz,2H),6.74–6.67(m,1H),6.20(dd,J=9.1,2.4Hz,1H) ,6.03(d,J=2.3Hz,1H),4.06(s,2H),3.91(dt,J=12.1,4.3Hz,2H),3.79–3.64(m,3H),3 .62–3.53(m,2H),3.37–3.27(m,6H),3.05(s,3H),2.09–1.95(m,6H),1.58–1.47(m,2H).

[0440] Embodiment 14

[0441] Preparation of (R)-N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 14

[0442]

[0443] Using the same preparation method as in Example 1 and using (R)-2-(2,5-difluorophenyl)pyrrolidine as a raw material, compound 13 can be obtained.

[0444] MS m / z(ESI):617[M+H] + .

[0445] 1 H NMR(400MHz,DMSO-d6)δ9.03(br,1H),8.49(br,1H),8.15(br,1H),7.92(s,1H),7.59(br,1H), 7.31–7.02(m,3H),6.83(br,1H),6.20(dd,J=8.9,2.2Hz,1H),6.10(d,J=2.3Hz,1H),5.34(br, 1H),3.94–3.85(m,1H),3.78(dt,J=11.6,3.9Hz,2H),3.68-3.53(m,2H),3.49–3.40(m,2H),3. 21(t,J=5.1Hz,4H),2.40(t,J=5.0Hz,4H),2.19(s,3H),2.02-1.81(m,6H),1.37–1.25(m,2H).

[0446] Embodiment 15

[0447] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(7-methyl-2,7-diazaspiro[3.5]nonan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 15

[0448]

[0449] Using the same preparation method as Example 12, with 7-methyl-2,7-diazaspiro[3.5]nonane as the raw material, compound 15 can be obtained.

[0450] MS m / z(ESI):601[M+1]+

[0451] 1 H NMR (400MHz, DMSO-d6) δ12.64(s,1H),10.03(s,1H),8.39(d,J=7.5Hz,1H),7.78(d,J=8.8Hz, 1H),7.47(s,1H),7.40(d,J=8.6Hz,1H),7.25(dd,J=8.6,1.6Hz,1H),7.05–6.93(m,3H),5.71 (dd,J=8.7,2.1Hz,1H),5.61(d,J=2.1Hz,1H),4.04(s,2H),3.82(dt,J=11.6,4.0Hz,2H),3.6 6(s,4H),3.61–3.41(m,3H),2.78(br,4H),2.56(s,3H),1.97-1.85(m,6H),1.43–1.33(m,2H).

[0452] Example 16

[0453] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(6-methyl-2,6-diazaspiro[3.3]heptane-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 16

[0454]

[0455] Using the same preparation method as Example 12, with 6-methyl-2,6-diazaspiro[3.5]heptane as raw material, compound 16 can be obtained.

[0456] MS m / z(ESI):573[M+1] +

[0457] 1 H NMR(400MHz,Chloroform-d)δ8.44(s,1H),8.19(d,J=7.4Hz,1H),7.74(s,1H),7.50 (d,J=8.7Hz,1H),7.33(d,J=8.6Hz,1H),7.16(d,J=8.6Hz,1H),6.75–6.67(m,2H),6 .61(tt,J=9.0,2.3Hz,1H),5.72(d,J=8.5Hz,1H),5.57(d,J=2.1Hz,1H),4.09–3.90 (m,8H),3.58-3.45(m,7H),2.42(s,3H),2.00(d,J=13.0Hz,2H),1.66-1.53(m,2H).

[0458] Embodiment 17

[0459] Preparation of N-(5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 17

[0460]

[0461]

[0462] Step 1 Synthesis of 5-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridine 17b

[0463] 5-Bromo-1H-pyrazolo[3,4-c]pyridine 17a (1.0 g, 5.0 mmol, 1.0 eq) was added to anhydrous THF (20 ml), and NaH (0.4 g, 10.0 mmol, 2.0 eq) was added in batches under ice-water bath cooling. After the addition was completed, the mixture was refluxed for 1 h, cooled to room temperature, and SEMCl (1.3 ml, 7.5 mmol, 1.5 eq) was added, and the reaction was continued for 2 h. A small amount of MeOH was added to quench the mixture, and the solvent was evaporated, and EA (50 ml) was added. The mixture was washed with H2O (15 ml) and brine (15 ml) in sequence, dried over anhydrous sodium sulfate, concentrated, and purified by flash column (PE~PE / EA=1:1) to obtain a yellow product 17b (560 mg).

[0464] MS m / z(ESI):330[M+1] +

[0465] Step 2 Synthesis of 5-(3,5-difluorobenzyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridine 17d

[0466] Zn powder (5.0 g, 76.5 mmol, 12.0 eq) was added to anhydrous THF (50 ml), 1,2-dibromoethane (5 drops) was added, N2 was purged 5 times, refluxed for 30 min, cooled to room temperature, TMSCl (4 drops) was added, and the reaction was continued for 30 min. 3,5-difluorobenzyl bromide (5.0 ml, 38.6 mmol, 1.0 eq) was added dropwise under ice-water cooling, and the temperature was naturally raised to room temperature. The reaction was allowed to proceed overnight to obtain 17c.

[0467] Compound 17b (200 mg, 0.61 mmol, 1.0 eq) was dissolved in anhydrous THF (15 ml), Pd(PPh3)4 (76 mg, 0.061 mmol, 0.1 eq) was added, N2 gas was purged 5 times, the reaction solution of compound 17c (about 10 ml) was added dropwise, after addition, reflux reaction was carried out for 1 h. The solvent was evaporated under reduced pressure, EA (30 ml) and H2O (15 ml) were added, the insoluble matter was filtered, the EA layer was separated, washed with brine, dried over anhydrous sodium sulfate, concentrated, and purified by flash column (PE~PE / EA=1:1) to obtain yellow oil 17d (210 mg).

[0468] MS m / z(ESI):376[M+1] +

[0469] Step 3 Synthesis of 5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-c]pyridine 17e

[0470] Compound 17d (400 mg, 1.1 mmol, 1.0 eq) was dissolved in 1,4-dioxane (30 ml), 4N HCl (20 ml, 20.0 mmol, 19.0 eq) was added, and the mixture was reacted at 70°C for 5 h. The mixture was evaporated to dryness under reduced pressure to obtain 17e, which was used directly in the next step without purification.

[0471] MS m / z(ESI):246[M+1] +

[0472] Step 4 Synthesis of 5-(3,5-difluorobenzyl)-3-iodo-1H-pyrazolo[3,4-c]pyridine 17f

[0473] 17e was dissolved in DMF (8 ml), and anhydrous potassium carbonate (0.54 g, 3.9 mmol, 3.0 eq) and iodine (0.69 g, 2.7 mmol, 2.0 eq) were added in sequence, and the mixture was reacted at room temperature for 2 h. EA (50 ml) was added for dilution, and the mixture was washed with H2O (15 ml) and brine (15 ml×2) in sequence, dried over anhydrous sodium sulfate, concentrated, and purified by flash column (PE~PE / EA=2:1) to obtain yellow solid 17f (220 mg).

[0474] MS m / z(ESI):372[M+1] +

[0475] Step 5 Synthesis of 17 g of 5-(3,5-difluorobenzyl)-3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridine

[0476] Compound 17f (220 mg, 0.59 mmol, 1.0 eq) was dissolved in THF (10 ml), and NaH (48 mg, 1.2 mmol, 2.0 eq) was added. The mixture was refluxed for 1 h and cooled to room temperature. SEMCl (0.12 g, 0.71 mmol, 1.2 eq) was added under ice-water cooling and the reaction was continued for 2 h. Flash column purification (PE-PE / EA=1:1) was performed to obtain 17 g of a yellow oil, which solidified after standing (150 mg).

[0477] MS m / z(ESI):502[M+1] +

[0478] Step 6 Synthesis of N-(5-(3,5-difluorobenzyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 17h

[0479] Compound 17g (130 mg, 0.26 mmol, 1.0 eq), 4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 9a (83 mg, 0.26 mmol, 1.0 eq), Cu (17 mg, 0.26 mmol, 1.0 eq), CuI (50 mg, 0.26 mmol, 1.0 eq), N,N'-dimethylethylenediamine (23 mg, 0.26 mmol, 1.0 eq) and K3PO4 (0.17 g, 0.78 mmol, 3.0 eq) were added to toluene (10 ml), purged with N2 gas 5 times, and refluxed for 2 h. The reaction solution was directly mixed and purified by flash column (DCM~DCM / MeOH=10:1) to obtain yellow solid 17h (120 mg).

[0480] MS m / z(ESI):692[M+1] +

[0481] Step 7: Synthesis of N-(5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 17.

[0482] Compound 17h (84 mg, 0.12 mmol, 1.0 eq) was dissolved in DCM (8 ml), TFA (2 ml) was added, the mixture was reacted at room temperature for 2 h, evaporated to dryness under reduced pressure, THF (8 ml) and H2O (2 ml) were added, LiOH (0.48 g, 20.0 mmol) was added, the mixture was reacted at room temperature overnight, H2O (20 ml) was added, the mixture was extracted with EA (50 ml×2), the organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, concentrated, and purified by preparative purification (DCM / MeOH=15:1) to obtain nearly white solid 17 (5 mg).

[0483] MS m / z(ESI):562[M+1] +

[0484] 1 H NMR(400MHz,Chloroform-d)δ8.87(s,1H),8.55(s,1H),8.11(d,J=7.4Hz,1H),7.77(s,1H),7 .52(d,J=9.0Hz,1H),6.86–6.74(m,2H),6.61(tt,J=9.1,2.5Hz,1H),6.21(dd,J=9.0,2.2Hz,1 H),6.09(d,J=2.3Hz,1H),4.23(s,2H),3.99(dt,J=11.9,3.9Hz,2H),3.65–3.50(m,3H),3.38( t,J=5.1Hz,4H),2.68(t,J=5.0Hz,4H),2.44(s,3H),2.02(d,J=5.1Hz,2H),1.65-1.57(m,2H).

[0485] Embodiment 18

[0486] Preparation of N-(6-(3,5-difluorobenzyl)imidazo[1,2-b]pyridazin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydropyran-4-yl)amino)-benzamide 18

[0487]

[0488] Using the same preparation method as Example 9 and taking 6-chloro-imidazo[1,2-b]pyridazine as the raw material, compound 18 can be obtained.

[0489] MS m / z(ESI):562[M+H] + .

[0490] 1H NMR (400 MHz, trifluoroacetate, Methanol-d4) δ7.91 (d, J = 9.4 Hz, 1H), 7.85–7.71 (m, 2H), 7.13 (d, J = 9.4 Hz, 1H), 7.01–6.92 (m, 2H), 6.85–6.75 (m, 1H), 6.38 (dd, J = 9.0, 2.4 Hz, 1H), 6.29 (d, J = 2.4 Hz, 1H), 4.23 (s, 2H), 3.92 (dt, J = 11.9, 4.0 Hz, 2H), 3.74–3.65 (m, 1H), 3.62 -3.57(m,2H),3.42-3.24(m,8H),2.94(s,3H),2.01(d,J=4.3Hz,2H),1.59–1.49(m,2H).

[0491] Embodiment 19

[0492] Preparation of N-(5-((4-(2-hydroxypropane-2-yl)piperidin-1-yl)methyl)-1H-indazol-3-yl)-4-(4-methylpiperidin-1-yl)-2-((tetrahydropyran-4-yl)amino)benzamide 19

[0493]

[0494]

[0495] The first step is the preparation of 2-fluoro-5-hydroxybenzonitrile 19a

[0496] Sodium triacetylborohydride (4 g, 18.9 mmol) was added to a dichloromethane solution (30 mL) of 2-fluoro-5-formylbenzocyanide I-1a (1 g, 6.7 mmol), and two drops of acetic acid were added, followed by stirring at room temperature overnight. Water (30 mL*3) was added for washing, dried over anhydrous Na2SO4, filtered, and concentrated to obtain the title compound 19a (930 mg, 6.2 mmol) with a yield of 91.8%.

[0497] MS m / z(ESI):152[M+H]+.

[0498] Step 2 Preparation of 2-fluoro-5-bromomethylbenzonitrile 19b

[0499] 19a (700 mg, 4.6 mmol) was dissolved in dichloromethane (20 mL), and phosphorus tribromide (800 mg, 2.96 mmol) was added under ice bath. After stirring at room temperature for 2 hours, the mixture was cooled and ice bathed. Water (20 mL) was added and stirring was continued for 30 minutes. The mixture was allowed to stand and the layers were separated. Anhydrous Na2SO4 was added to the organic phase, which was dried, filtered and concentrated to give the crude title compound 19b which was used directly in the next step.

[0500] Step 3 Preparation of 2-fluoro-5-((4-(2-hydroxypropane-2-yl)piperidin-1-yl)methyl)benzonitrile 19d

[0501] Compound 19b (985 mg, 4.6 mmol) was dissolved in dichloromethane (20 mL), and diisopropylethylamine (2 g, 15.5 mmol) and 2-(4-piperidinyl)-2-propanol 19c (700 mg, 4.89 mmol) were added, and the mixture was stirred at room temperature overnight. The mixture was concentrated and purified by flash column chromatography (DCM / MeOH, 0-10%) to obtain the title compound 19d (510 mg, 1.85 mmol) with a yield of 39.9%.

[0502] MS m / z(ESI):277[M+H]+.

[0503] Step 4 Preparation of 2-(1-((3-amino-1-hydroindazol-5-yl)methyl)piperidin-4-yl)-2-propanol 19e

[0504] Compound 19d (510 mg, 1.85 mmol) was dissolved in ethanol (10 mL), hydrazine hydrate (10 mL) was added, and the mixture was heated to 100°C in a microwave oven for 2 hours and then concentrated and purified by Flash column chromatography (DCM / MeOH, 0-25%) to give the title compound 19e (320 mg, 1.11 mmol) in a yield of 60.1%.

[0505] MS m / z(ESI):289[M+H]+.

[0506] Step 5 Preparation of N-(5-((4-(2-hydroxypropane-2-yl)piperidin-1-yl)methyl)-1H-indazol-3-yl)-4-(4-methylpiperidin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydropyran-4-yl)acetamido)benzamide 19f

[0507] Compound 19e (836 mg, 1.93 mmol) was dissolved in dichloromethane (20 mL), and 1f (320 mg, 1.11 mmol) and diisopropylethylamine (500 mg, 3.88 mmol) were added under ice bath. After reacting at room temperature for 2 hours, the mixture was concentrated and purified by thin plate chromatography (DCM / MeOH, 25%) to obtain the title compound 19f (120 mg, 0.17 mmol) with a yield of 15.8%.

[0508] MS m / z(ESI):686[M+H]+.

[0509] Step 7 Preparation of N-(5-((4-(2-hydroxypropane-2-yl)piperidin-1-yl)methyl)-1H-indazol-3-yl)-4-(4-methylpiperidin-1-yl)-2-((tetrahydropyran-4-yl)amino)benzamide 19

[0510] Compound 19f (120 mg, 0.17 mmol) was dissolved in methanol (10 mL), potassium carbonate (1 g, 7.2 mmol) was added, and the mixture was stirred at room temperature overnight. The mixture was concentrated and purified by thin-plate chromatography (DCM / MeOH, 25%) to give the title compound 19 (10 mg, 0.017 mmol) in a yield of 9.7%.

[0511] MS m / z(ESI):590[M+H]+.

[0512] 1 H NMR (400MHz, DMSO-d6) δ12.62(s,1H),10.08(s,1H),8.25(d,J=7.7Hz,1H),7.76(d,J=9.1Hz,1H ),7.49–7.32(m,2H),7.27(d,J=8.6Hz,1H),6.25–6.16(m,1H),6.10(d,J=2.3Hz,1H),4.00(s,1H ),3.75(m,2H),3.64-3.45(m,5H),3.23(t,J=5.1Hz,4H),2.87(d,J=10.3Hz,2H),2.41(t,J=5.0 Hz, 4H), 2.19 (s, 3H), 1.87-1.75 (m, 3H), 1.59 (d, J = 11.7Hz, 2H), 1.30-1.19 (m, 4H), 0.96 (s, 6H).

[0513] Embodiment 20

[0514] Preparation of N-(5-(3,5-difluorophenyl)-1H-indazol-3-yl)-4-(5-(dimethylamino)hexahydrocyclopenta[c]pyrrol-2(1H)-yl)-2((tetrahydro-2H-pyran-4-yl)amino)benzamide 20

[0515]

[0516] By adopting the same preparation method as Example 12, with N,N-dimethyloctahydrocyclopenta[c]pyrrole-5-amine as the raw material, compound 20 can be obtained.

[0517] MS m / z(ESI):615[M+1] + .

[0518] 1H NMR (400MHz, DMSO-d6) δ12.58(s,1H),9.96(s,1H),8.32(d,J=7.5Hz,1H),7.73(d,J=9.0Hz,1H),7.44(s,1H),7.36(d, J=8.6Hz,1H),7.20(dd,J=8.6,1.6Hz,1H),7.04–6.88(m,3H),5.88(dd,J=8.9,2.1Hz,1H),5.74(d,J=2.2Hz,1H),4.00 (s,2H),3.77(dt,J=11.6,3.9Hz,2H),3.65-3.55(m,1H),3.50–3.40(m,2H),3.39–3.35(m,2H),3.19(dd,J=10.2,3.0H z,2H),2.65(d,J=11.8Hz,2H),2.48-2.37(m,1H),2.14-2.03(s,8H),1.91(dd,J=13.3,3.4Hz,2H),1.36-1.16(m,6H).

[0519] Embodiment 21

[0520] Preparation of N-(5-(1-(3,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 21

[0521]

[0522]

[0523] The first step is the preparation of 5-(1-(3,5-difluorophenyl)-2-hydroxyethyl)-2-fluorophenylacetonitrile 21a

[0524] Compound 10b (2 g, 6 mmol), zinc cyanide (702 mg, 6 mmol) and tetrakistriphenylphosphine palladium (693 mg, 0.6 mmol) were mixed in DMF (8 mL), replaced with nitrogen twice, and the reaction solution was reacted at 110 ° C in a microwave for 1 hour. After cooling, the reaction solution was poured into water (30 mL) and extracted with ethyl acetate (50×2 mL). The organic phase was dried and spin-dried to obtain a crude product, which was purified by column chromatography (PE / EA=30:1) to obtain compound 21a (1.5 g, 5.4 mmol) with a yield of 90%.

[0525] MS m / z(ESI):278[M+1] + .

[0526] Step 2 Preparation of 5-(1-(3,5-difluorophenyl)-2-fluoroethyl)-2-fluorophenylacetonitrile 21b

[0527] Compound 21a (1.5 g, 5.41 mmol) was dissolved in dichloromethane (30 mL), and the solution was cooled to 0°C. DAST (4.36 g, 27.0 mmol) was slowly added dropwise to the solution. The reaction solution was stirred at room temperature overnight, and then the reaction solution was slowly added dropwise to ice water (30 mL), extracted with dichloromethane (50 mL), and the organic phase was dried and spin-dried to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 21b (245 mg).

[0528] MS m / z(ESI):280[M+H] + .

[0529] Step 3 Preparation of 5-(1-(3,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-amine 21c

[0530] Compound 21b (245 mg, 0.88 mmol) and hydrazine hydrate (2.2 g, 44 mmol) were dissolved in ethanol (6 mL). The reaction solution was reacted at 90 ° C. in a microwave reactor for 4 hours, and the crude product was spin-dried to obtain a crude product. After purification by silica gel column chromatography (DCM / MeOH=15:1), compound 21c (94 mg) was obtained with a yield of 37%.

[0531] MS m / z(ESI):292[M+H] + .

[0532] Step 4 Preparation of N-(5-(1-(3,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 21d

[0533] Compound 1f (70 mg, 0.16 mmol) was dissolved in dichloromethane (10 mL) and added dropwise to a solution of compound 21c (48 mg, 0.16 mmol) and DIPEA (63 mg, 0.48 mmol) in tetrahydrofuran (10 mL) cooled to -20 ° C. The reaction was stirred at -20 ° C for 2 hours, and then continued to stir at room temperature for 2 hours. The reaction solution was diluted with dichloromethane (50 mL), washed with saturated brine, dried and spin-dried to give a crude product, which was purified by column chromatography (DCM / MeOH=15:1) to give compound 21d (370 mg, 0.48 mmol) with a yield of 45%.

[0534] MS m / z(ESI):689[M+H] + .

[0535] Step 5 Preparation of N-(5-(1-(3,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 21

[0536] Compound 21d (77 mg, 0.11 mmol) was dissolved in a mixed solvent of methanol (8 mL) and water (8 mL), potassium carbonate (77 mg, 0.55 mmol) was added, the reaction solution was stirred at room temperature for 3 hours, water (10 ml) was added, and it was extracted with dichloromethane (50 ml). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by silica gel column chromatography (DCM / MOH=15:1) to obtain compound 21 (25 mg, 0.042 mmol), with a yield of 38%.

[0537] MS m / z(ESI):593[M+1]+.

[0538] 1 H NMR (400MHz, DMSO-d6) δ12.70(s,1H),10.12(s,1H),8.31(d,J=7.7Hz,1H),7.77(d,J=9.1Hz,1H ),7.52(s,1H),7.45-7.23(m,3H),7.07(t,J=8.6Hz,2H),6.21(dd,J=9.0,2.3Hz,1H),6.11(d,J= 2.3Hz,1H),5.10(d,J=7.9Hz,1H),4.98(d,J=7.9Hz,1H),4.89-4.79(m,1H),3.84-3.60(m,3H), 3.60-3.40(m,2H),3.34(s,4H),2.59(s,4H),2.32(s,3H),2.00-1.82(m,2H),1.24-1.19(m,2H).

[0539] Embodiment 22

[0540] Preparation of 5-(3,5-difluorophenyl)-N-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)-1H-indazole-3-carboxamide 22

[0541]

[0542] Step 1 Preparation of 1-(3-methoxy-4-nitrophenyl)-4-methylpiperazine 22b

[0543] Compound 22a (5.1 g, 30 mmol) and N-methylpiperazine (6 g, 60 mmol) were dissolved in N-methylpyrrolidone (40 ml), and the solution was stirred at 100 °C for 2 hours. After cooling, it was poured into ice water (200 mL), and the solution was extracted with ethyl acetate (150 mL). The organic phase was washed with saturated brine, dried and spin-dried to obtain a crude product, which was purified by column chromatography to obtain compound 22b (6 g).

[0544] MS m / z(ESI):252[M+1] + .

[0545] Step 2 Preparation of 2-methoxy-4-(4-methylpiperazin-1-yl)aniline 22c

[0546] Compound 22b (6 g, 23.9 mmol) was dissolved in methanol (100 mL), palladium carbon (0.6 g) was added, and the reaction solution was stirred for 2 hours under a hydrogen atmosphere. The reaction solution was filtered and dried to obtain compound 22c (4.5 g, 20 mmol) with a yield of 85%.

[0547] MS m / z(ESI):222[M+1] + .

[0548] Step 3 Preparation of di-tert-butyl 5-bromo-1H-indazole-1,3-dicarboxylate 22e

[0549] To a solution of compound 22d (1 g, 4.1 mmol) and Boc2O (2.7 g, 12.4 mmol) in dichloromethane (10 mL) were added tert-butyl alcohol (10 mL) and DMAP (0.5 g, 4 mmol), and the system was stirred at room temperature overnight. The reaction solution was spin-dried, silica gel was added to mix the sample, and column purification was performed to obtain compound 22e (1.5 g, 3.73 mmol), with a yield of 91%.

[0550] MS m / z(ESI):397[M+1] + .

[0551] Step 4 Preparation of tert-butyl 5-bromo-1H-indazole-3-carboxylate 22f

[0552] K2CO3 (2 g, 14.5 mmol) was added to a solution of compound 22e (1.1 g, 2.8 mmol) in methanol (30 mL), and the system was stirred at room temperature overnight. Water (200 mL) was added to the reaction solution and the stirring was continued for 15 minutes, and the solution was filtered, washed with water, and dried to obtain compound 22f (0.8 g, 2.7 mmol) with a yield of 97%.

[0553] MS m / z(ESI):297[M+1] + .

[0554] Step 5 Preparation of 22 g of tert-butyl 5-bromo-1-trityl-1H-indazole-3-carboxylate

[0555] Triethylamine (0.55 g, 5.4 mmol) and trichlorobenzene (0.9 g, 3.2 mmol) were added to a dichloromethane (20 mL) solution of compound 22f (0.8 g, 2.7 mmol), and the system was stirred at room temperature overnight. The reaction solution was spin-dried, silica gel was added to mix the sample, and column purification was performed to obtain compound 22g (1.2 g, 2.2 mmol) with a yield of 83%.

[0556] MS m / z(ESI):539[M+1] + .

[0557] Step 6 Preparation of tert-butyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-trityl-1H-indazole-3-carboxylate 22h

[0558] A DMF (5 mL) solution of compound 22g (100 mg, 0.18 mmol), pinacol borate (100 mg, 0.39 mmol), Pd(dppf)Cl2 (30 mg, 0.04 mmol) and potassium acetate (50 mg, 0.5 mmol) was heated to 100°C for 3 hours, cooled to room temperature, filtered, diluted with ethyl acetate (50 mL), washed with saturated brine, dried, filtered and concentrated to give compound 22h (100 mg, 0.17 mmol) in a yield of 92%.

[0559] MS m / z(ESI):587[M+1] + .

[0560] Step 7 Preparation of tert-butyl 5-(3,5-difluorophenyl)-1-trityl-1H-indazole-3-carboxylate 22i

[0561] A toluene (10 mL) solution of compound 22h (500 mg, 0.85 mmol), 1-(bromomethyl)-3,5-difluorobenzene (500 mg, 2.41 mmol), Pd(PPh3)4 (200 mg, 0.17 mmol) and potassium phosphate (500 mg, 2.36 mmol) was heated to 100°C under nitrogen protection for 3 hours, then cooled to room temperature, filtered, diluted with ethyl acetate (50 mL), washed with saturated brine, dried, filtered, and concentrated to give a crude product, which was purified by silica gel column chromatography to give compound 22i (200 mg, 0.34 mmol) with a yield of 40%.

[0562] MS m / z(ESI):587[M+1] + .

[0563] Step 8 Preparation of tert-butyl 5-(3,5-difluorophenyl)-1-trityl-1H-indazole-3-carboxylate 22j

[0564] Trifluoroacetic acid (2 mL) was added to a solution of compound 22i (200 mg, 0.34 mmol) in dichloromethane (8 mL). The reaction solution was stirred at room temperature for 2 hours. The reaction solution was concentrated and dried to give compound 22j (60 mg, 0.20 mmol) in a yield of 61%.

[0565] MS m / z(ESI):307[M+1] + .

[0566] Step 9 Preparation of tert-butyl 5-(3,5-difluorophenyl)-N-(2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)-1H-indazole-3-carboxamide 22

[0567] Compound 22c (203 mg, 0.92 mmol) and compound 22j (220 mg, 0.76 mmol) were dissolved in dichloromethane (15 mL), HATU (433 mg, 1.14 mmol) and DIPEA (44 mg, 1.52 mmol) were added, and the reaction solution was stirred at room temperature for 2 hours. The reaction solution was diluted with dichloromethane (30 mL), washed with saturated brine, and the organic phase was dried and spin-dried to obtain a crude product, which was purified by column chromatography and then by reverse phase column purification to obtain compound 22 (55 mg, 0.11 mmol) with a yield of 15%.

[0568] MS m / z(ESI):492[M+1] + .

[0569] 1 H NMR (400MHz, DMSO-d6) δ13.66(s,1H),9.26(s,1H),8.14(d,J=8.8Hz,1H),8.06(s,1H),7.63–7.52(m,1H),7.34(dd,J=8.7,1.6Hz,1H), 7.09–6.90(m,3H),6.68(d,J=2.5Hz,1H),6.50(dd,J=8.9,2.5Hz,1H),4.10(s,2H),3.88(s,3H),3.17(s,4H),2.62(s,4H),2.33(s,3H).

[0570] Embodiment 23

[0571] Preparation of N-(5-(1-(3,5-difluorophenyl)-2-methoxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 23

[0572]

[0573] The first step is the preparation of 2-bromo-4-(1-(3,5-difluorophenyl)-2-methoxyethyl)-1-fluorobenzene 23a

[0574] Compound 10b (400 mg, 1.2 mmol) was dissolved in tetrahydrofuran (15 mL), cooled to 0°C, sodium hydroxide (58 mg, 1.44 mmol) was added, and the reaction solution was stirred at 0°C for 1 hour. Methyl iodide (198.8 mg, 1.44 mmol) was added to the reaction solution, and the reaction solution was stirred at room temperature overnight. The reaction solution was quenched with saturated ammonium chloride solution, extracted with ethyl acetate (30 mL), dried and spin-dried to give a crude product, which was purified by column chromatography to give compound 23a (330 mg, 0.96 mmol) with a yield of 80%.

[0575] MS m / z(ESI):345[M+H] + .

[0576] Step 2 Preparation of 5-(1-(3,5-difluorophenyl)-2-methoxyethyl)-2-fluorobenzonitrile 23b

[0577] Compound 23a (330 mg, 0.96 mmol), zinc cyanide (112 mg, 0.96 mmol) and tetrakistriphenylphosphine palladium (115 mg, 0.1 mmol) were mixed in DMF (4 mL), replaced with nitrogen twice, and the reaction solution was reacted at 110 ° C. on a microwave for 1 hour. After cooling, the reaction solution was poured into water (30 mL) and extracted with ethyl acetate (50 mL). The organic phase was dried and spin-dried to obtain a crude product, which was purified by column chromatography (PE / EA=30:1) to obtain compound 23b (200 mg, 0.69 mmol) with a yield of 72%.

[0578] MS m / z(ESI):292[M+H] + .

[0579] Step 3 Preparation of 5-(1-(3,5-difluorophenyl)-2-methoxyethyl)-1H-indazol-3-amine 23c

[0580] Compound 23b (200 mg, 0.69 mmol) and hydrazine hydrate (345 mg, 6.9 mmol) were mixed in N-methylpyrrolidone (6 mL), and the reaction solution was reacted at 90 ° C in a microwave reactor for 4 hours. After cooling, it was added to (30 mL) and extracted with ethyl acetate (50 mL). The organic phase was dried and spin-dried to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 23c (194 mg, 0.64 mmol).

[0581] MS m / z(ESI):304[M+H] + .

[0582] Step 4 Preparation of N-(5-(1-(3,5-difluorophenyl)-2-methoxy)ethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 23d

[0583] Compound 1f (100 mg, 0.33 mmol) was dissolved in tetrahydrofuran (5 mL), added to a solution of compound 23c (174.6 mg, 0.33 mmol) and DIPEA (85 mg, 0.66 mmol) in tetrahydrofuran (15 mL), and the reaction was stirred at -20 ° C for 2 hours, and then continued to stir at room temperature for 2 hours. The reaction solution was diluted with dichloromethane (50 mL), washed with saturated brine, dried and spin-dried to give a crude product, which was purified by column chromatography (DCM / MeOH=10:1) to give compound 23d (140 mg, 0.2 mmol) with a yield of 60%.

[0584] MS m / z(ESI):701[M+H] + .

[0585] Step 5 Preparation of N-(5-(1-(3,5-difluorophenyl)-2-methoxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 23

[0586] Compound 23d (140 mg, 0.2 mmol) was dissolved in a mixed solvent of methanol (10 mL) and water (5 mL), potassium carbonate (82.8 mg, 0.6 mmol) was added, the reaction solution was stirred at room temperature for 3 hours, water (20 ml) was added, and it was extracted with dichloromethane (30 ml). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 23 (20 mg, 0.033 mmol), with a yield of 17%.

[0587] MS m / z(ESI):605[M+H] + .

[0588] 1 H NMR (400MHz, DMSO-d6) δ12.63(s,1H),10.09(s,1H),8.32(d,J=7.7Hz,1H),7.76(d,J=9.0Hz,1H),7.49(s,1H ),7.37(d,J=8.7Hz,1H),7.34–7.19(m,2H),7.01(t,J=8.8Hz,2H),6.20(dd,J=9.0,2.3Hz,1H),6.10(d,J=2.4 Hz,1H),4.67(t,J=8.0Hz,1H),3.99(dt,J=28.0,8.9Hz,2H),3.86–3.73(m,2H),3.73–3.59(m,1H),3.54–3.41 (m,2H),3.24(d,J=2.6Hz,7H),2.40(t,J=5.0Hz,4H),2.19(s,3H),1.92(d,J=13.2Hz,2H),1.36-1.28(m,2H).

[0589] Embodiment 24

[0590] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-methyl-7-((tetrahydro-2H-pyran-4-yl)amino)-1,2,3,4-tetrahydroisoquinoline-6-carboxamide 24

[0591]

[0592] Step 1-(6-Bromo-1,2,3,4-tetrahydroisoquinolin-2-yl)-2,2,2-trifluoroethane-1-one 24b

[0593] 24a (2.12 g, 10 mmol) and 10 mL of trifluoroacetic anhydride were added to a dry reaction bottle, and stirred at room temperature for 1 hour. TLC showed that the raw materials had reacted completely. The reaction solution was concentrated under reduced pressure, diluted with ethyl acetate, and washed with cold sodium bicarbonate solution 3 times. The organic phase was separated and dried over anhydrous sodium sulfate, and then concentrated under pressure. The obtained oil 24b was directly used for the next reaction.

[0594] MS m / z(ESI):308[M+H]+

[0595] Step 2 Preparation of 1-(6-bromo-7-nitro-1,2,3,4-tetrahydroisoquinolin-2-yl)-2,2,2-trifluoroethane-1-one 24c

[0596] Add 50 mL of concentrated sulfuric acid to the reaction flask, cool to 0-5°C, add compound 24b (3.1 g, 10 mmol), and then add potassium nitrate (1.2 g, 12.0 mmol) in batches, keeping the internal temperature below 10°C. Stir overnight after addition. TLC shows that the raw materials have reacted. Pour the reaction solution into ice water, precipitate solids, filter, and obtain crude product 24c of sufficient purity for the next step reaction.

[0597] MS m / z(ESI):353[M+H]+

[0598] Step 3 Preparation of 7-nitro-2-(2,2,2-trifluoroacetyl)-1,2,3,4-tetrahydroisoquinoline-6-carbonitrile 24d

[0599] Add 24c (353 mg, 1.0 mmol), CuCN (180 mg, 2.0 mmol), and DMF (5 mL) to a microwave reaction bottle. The mixture was heated in a microwave at 130 ° C for 1 hour. TLC showed that the raw materials were basically reacted. The reaction solution was diluted with a large amount of ethyl acetate, filtered, and the filtrate was washed with ammonium chloride solution. The organic phase was dried over anhydrous sodium sulfate and concentrated to a small volume under reduced pressure. An appropriate amount of petroleum ether was added to slurry, and the crude product 24d obtained by filtration was directly used for the next step reaction.

[0600] MS m / z(ESI):300[M+H]+

[0601] Step 4 Preparation of 7-nitro-1,2,3,4-tetrahydroisoquinoline-6-carbonitrile 24e

[0602] 24d (300 mg, 1.0 mmol) was dissolved in 20 ml of methanol, and 10 ml of saturated potassium carbonate solution was added. The mixture was stirred for half an hour. TLC showed that the reaction of the raw material was complete. Most of the methanol in the reaction solution was concentrated under reduced pressure, and ethyl acetate and water were added to separate the organic phase. The organic phase was dried over anhydrous sodium sulfate and flash purified (dichloromethane: methanol = 20: 1) to obtain 24e (142 mg, 0.7 mmol, 70%).

[0603] MS m / z(ESI):204[M+H]+

[0604] Step 5 Preparation of 2-methyl-7-nitro-1,2,3,4-tetrahydroisoquinoline-6-carbonitrile 24f

[0605] Compound 24e (1.0 g, 5 mmol) was dissolved in 50 ml of dichloromethane, and paraformaldehyde (1.5 g, 50 mmol) and 5 ml of acetic acid were added, followed by NaBH(OAc)3 (2.12 g, 10 mmol). Stirring was continued at room temperature overnight, and TLC showed that most of the raw materials had reacted. Aqueous potassium carbonate solution was added to quench the reaction, and the organic phase was separated, and silica gel was added to mix the sample, and flash purification (dichloromethane: methanol = 20: 1) was performed to obtain 24f (650 mg, 3 mmol, 60%).

[0606] MS m / z(ESI):218[M+H]+

[0607] Step 6 Preparation of 24 g of 2-methyl-7-nitro-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid

[0608] Add 24f (650 mg, 3 mmol), 10 ml of hydrogen bromide solution (50% wt), and 5 ml of acetic acid to a microwave reaction bottle, and then heat in a microwave at 120°C for 1 hour. TLC shows that most of the raw materials have reacted. After the reaction solution is left at room temperature overnight, a large amount of solid precipitates, and the solid obtained by filtration is 24g (350 mg, 1.5 mmol, 50%).

[0609] MS m / z(ESI):237[M+H]+

[0610] Step 7 Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-methyl-7-nitro-1,2,3,4-tetrahydroisoquinoline-6-carboxamide 24h

[0611] Add compound 24g (100mg, 0.42mmol), dichloromethane and 1ml oxalyl chloride to a dry reaction bottle. Stir at room temperature for 4 hours, then concentrate under reduced pressure, dry with dichloromethane, add dry tetrahydrofuran and 0.2ml DIPEA, and cool to -40~-50℃ under nitrogen protection. Then slowly add a tetrahydrofuran solution of 5-(3,5-difluorobenzyl)-1H-indazole-3-amine (109mg, 0.42mmol). After the addition is complete, the temperature rises to room temperature naturally. TLC shows that most of the raw materials have reacted. Add silica gel to mix the sample, and flash purify (dichloromethane: methanol = 15:1) to obtain 24h (60mg, 0.12mmol, 28%).

[0612] MS m / z(ESI):478[M+H]+

[0613] Step 8 Preparation of 7-amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-methyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamide 24i

[0614] Add 24h (60 mg, 0.12 mmol), 5 ml tetrahydrofuran and 0.5 ml water to the reaction flask. Then add 65 mg reduced iron powder. After the addition, reflux at 60 ° C for 2 hours. TLC shows that the raw material has reacted. Add a large amount of ethyl acetate to dilute the reaction solution, filter the filtrate and purify it with flash (dichloromethane: methanol = 15: 1) to obtain 24i (30 mg, 0.067 mmol, 50%).

[0615] MS m / z(ESI):448[M+H]+

[0616] Step 9 Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-methyl-7-((tetrahydro-2H-pyran-4-yl)amino)-1,2,3,4-tetrahydroisoquinoline-6-carboxamide 24

[0617] Compound 24i (30 mg, 0.067 mmol) was dissolved in dichloromethane, and I-2 (100 mg, 1.0 mmol) and 0.2 ml of acetic acid were added. Then NaBH(OAc)3 (21.2 mg, 0.1 mmol) was added. Stir at room temperature overnight. Potassium carbonate solution was added, and the organic phase was separated. After concentration under reduced pressure, it was purified by preparative thin layer chromatography (dichloromethane: methanol = 10: 1) to obtain 24 (10 mg, 0.0188 mmol, 30%).

[0618] MS m / z(ESI):532[M+H]+

[0619] 1 H NMR (400MHz, DMSO-d6) δ12.70(s,1H),10.39(s,1H),7.67(s,1H),7.59(d,J=7.6H z,1H),7.51(s,1H),7.42(d,J=8.6Hz,1H),7.26(dd,J=8.5,1.6Hz,1H),7.06–6.9 1(m,3H),6.52(s,1H),4.04(s,2H),3.86-3.77(m,2H),3.62-3.39(m,5H),2.78-2 .70(m,2H),2.68-2.56(m,2H),2.35(s,3H),1.97-1.86(m,2H),1.38-1.27(m,2H).

[0620] Embodiment 25

[0621] Preparation of 4-((1R,3S,5s,7s)-5-aminoadamantan-2-yl)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide hydrochloride 25

[0622]

[0623] The first step is the preparation of ethyl 2-bromo-4-iodobenzoate 25b

[0624] 25a (3.28 g, 10 mmol) and 50 ml of DCM were added to a dry reaction bottle, and 10 ml of oxalyl chloride was added after the addition was complete. The mixture was stirred at room temperature for 1 hour after the addition was complete. TLC showed that the raw materials had reacted completely. The reaction solution was concentrated under reduced pressure, diluted with ethyl acetate, and washed with sodium bicarbonate solution 3 times. The organic phase was separated and dried over anhydrous sodium sulfate, and then concentrated under pressure. The obtained oily product 25b was directly used for the next step reaction.

[0625] MS m / z(ESI):357[M+H] +

[0626] Step 2 Preparation of benzyl (1s, 3R, 5S, 7s)-4-(3-bromo-4-(ethoxycarbonyl)phenyl)-4-hydroxyadamantane-1-carboxylate 25d

[0627] Add 25b (350 mg, 1.0 mmol) and 10 ml of tetrahydrofuran to the reaction flask, cool to -20 °C and then add i-PrMgCl (1.0 ml, 1 M), stir at this temperature for 0.5 hours, add 25c (284 mg, 1.0 mmol), and naturally warm overnight. TLC shows that the product is generated. Add ammonium chloride solution to quench the reaction. Add ethyl acetate to extract, and wash the organic phase with brine. After the organic phase is separated, add silica gel and mix the sample, flash purify (PE:EA=10:1) to obtain 25d (250 mg, 0.5 mmol, 50%).

[0628] MS m / z(ESI):497[M-OH] +

[0629] Step 3 Preparation of benzyl (1s, 3R, 5S, 7s)-4-(3-bromo-4-(ethoxycarbonyl)phenyl)adamantane-1-carboxylate 25e

[0630] Add 25d (500 mg, 1.0 mmol), 5 ml trifluoroacetic acid, and 1 ml Et3SiH to a single-mouth bottle. Stir at room temperature for 2 hours. TLC shows that the raw material has reacted completely. Concentrate most of the trifluoroacetic acid under reduced pressure, add DCM silica gel to mix the sample, and flash purify (PE:EA=10:1) to obtain 25e (495 mg, 1.0 mmol, 100%).

[0631] MS m / z(ESI):497[M+H] +

[0632] Step 4 Preparation of benzyl (1s, 3R, 5S, 7s)-4-(3-((tert-butoxycarbonyl)amino)-4-(ethoxycarbonyl)phenyl)adamantane-1-carboxylate 25f

[0633] Mix 25e (495 mg, 1.0 mmol), cesium carbonate (646 mg, 2.0 mmol), palladium acetate (2.2 mg, 0.1 mmol), XantPhos (5.78 mg, 0.1 mmol), tert-butyl carbamate (117 mg, 1.0 mmol), and add 10 ml of dioxane. Stir at 100 ° C for 1 hour. TLC shows that the raw materials have reacted. Add ethyl acetate to dilute the reaction solution and filter. The filtrate is flash purified (PE: EA = 10: 1) to obtain 25f (265 mg, 0.5 mmol, 50%).

[0634] MS m / z(ESI):534[M+H] +

[0635] Step 5 Preparation of 25 g of (1s, 3R, 5S, 7s)-4-(3-((tert-butoxycarbonyl)amino)-4-(ethoxycarbonyl)phenyl)adamantane-1-carboxylic acid

[0636] Compound 25f (1.06 g, 2.0 mmol) was dissolved in tetrahydrofuran, wet palladium carbon (500 mg) was added, and stirred overnight in a hydrogen atmosphere (hydrogen balloon). TLC showed that the raw material had reacted completely, and the mixture was filtered through celite, and the filtrate was concentrated to obtain 25 g (600 mg, 68%).

[0637] MS m / z(ESI):444[M+H] +

[0638] Step 6 Preparation of ethyl 4-((1R,3S,5s,7s)-5-(((benzyloxy)carbonyl)amino)adamantan-2-yl)-2-((tert-butoxycarbonyl)amino)benzoate 25h

[0639] Add 25g (444mg, 1.0mmol), 10ml toluene, DDPA (275mg, 1.0mmol), benzyl alcohol (216mg, 2.0mmol) and triethylamine (200mg, 2.0mmol) to a dry reaction bottle. The mixture was reacted at 120℃ for 4 hours. TLC showed that the raw materials were basically reacted. Ethyl acetate was added to dilute the reaction solution, washed with sodium bicarbonate, and the organic phase was separated and flash purified (PE:EA=15:1) to obtain 25h (400mg, 73%).

[0640] MS m / z(ESI):549[M+H] +

[0641] Step 7 Preparation of ethyl 2-amino-4-((1R,3S,5s,7s)-5-(((benzyloxy)carbonyl)amino)adamantan-2-yl)benzoate 25i

[0642] Compound 25h (548 mg, 1.0 mmol), 20 ml of dichloromethane, and then 2 ml of trifluoroacetic acid were added to a dry reaction bottle. After the addition, the mixture was stirred at room temperature overnight. TLC showed that the raw materials had reacted completely. The reaction solution was concentrated under reduced pressure to remove most of the trifluoroacetic acid and dichloromethane, and then diluted with ethyl acetate, washed with sodium bicarbonate solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 25i (450 mg, 100%).

[0643] MS m / z(ESI):449[M+H] +

[0644] Step 8 Preparation of ethyl 4-((1R,3S,5s,7s)-5-(((benzyloxy)carbonyl)amino)adamantan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzoate 25j

[0645] Tetrahydro-4H-pyran-4-one (200 mg, 2.0 mmol), 25i (450 mg, 1.0 mmol), dichloromethane, acetic acid (1.0 ml) and NaBH(OAc)3 (636 mg, 3.0 mmol) were added to the reaction bottle. The mixture was stirred at room temperature overnight, and then sodium bicarbonate was added to quench the reaction. After the organic phase was separated, silica gel was added to mix the sample, and flash purified (PE / EA=10:1) 25j (370 mg, 0.7 mmol, 70%).

[0646] MS m / z(ESI):533[M+H] +

[0647] Step 9 Preparation of 4-((1R,3S,5s,7s)-5-(((benzyloxy)carbonyl)amino)adamantan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzoic acid 25k

[0648] Compound 25j (532 mg, 1.0 mmol) was dissolved in 10 ml of tetrahydrofuran, and 2 ml of lithium hydroxide solution (10%) was added. The mixture was stirred at room temperature for 2 hours. TLC showed that the reaction of the raw material was complete. Most of the tetrahydrofuran was removed under reduced pressure, and an appropriate amount of water was added to dilute the residue. An appropriate amount of citric acid was added to neutralize the reaction solution, and ethyl acetate was added to extract for more than 3 times. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 25k (400 mg, 0.8 mmol, 80%).

[0649] MS m / z(ESI):505[M+H] +

[0650] Step 10 Preparation of 4-((1R,3S,5s,7s)-5-(((benzyloxy)carbonyl)amino)adamantan-2-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoic acid 25l

[0651] Add 25k (252 mg, 0.5 mmol), 0.5 ml trifluoroacetic anhydride and dichloromethane to a dry reaction flask. After stirring at room temperature for 1 hour, TLC shows that the raw materials have reacted. Add water and stir for half an hour, separate the organic phase, dry it with anhydrous sodium sulfate, and concentrate under reduced pressure to obtain 25l (300 mg, 0.5 mmol, 100%).

[0652] MS m / z(ESI):601[M+H] +

[0653] Step 11 Preparation of benzyl ((1s, 3R, 5S, 7s)-4-(4-((5-(3,5-difluorobenzyl)-1H-indazol-3-yl)carboxamide)-3-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)-acetylamino)phenyl)adamantan-1-yl)carbamate 25m

[0654] Add 25l (60mg, 0.1mmol), 5ml dichloromethane and 1ml oxalyl chloride to a dry reaction bottle. Stir at room temperature for 4-6 hours, concentrate the reaction solution to dryness under reduced pressure, dilute with tetrahydrofuran, cool to -40°C, and protect with nitrogen. Then add 0.2ml DIPEA, and then slowly add a THF solution of I-2 (26mg, 0.1mmol). After the addition, naturally warm to room temperature. TLC shows that most of the raw materials have reacted. Concentrate the reaction solution to an appropriate volume, and then purify it by preparative thin layer chromatography (DCM: MeOH = 10: 1) to obtain 25m (42mg, 0.05mmol, 50%).

[0655] MS m / z(ESI):842.9[M+H] +

[0656] Step 12 Preparation of benzyl((1s,3R,5S,7s)-4-(4-((5-(3,5-difluorobenzyl)-1H-indazol-3-yl)carboxamide)-3-((tetrahydro-2H-pyran-4-yl)amine)phenyl)adamantan-1-yl)carbamate 25n

[0657] Add 25m (42 mg, 0.05 mmol) and 3 ml of methanol to the reaction flask, add 0.5 ml of potassium carbonate solution after dissolving, TLC shows that the raw material has reacted completely after 10 minutes, concentrate under reduced pressure to remove methanol, add ethyl acetate to extract, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain 25n (30 mg, 0.04 mmol, 80%).

[0658] MS m / z(ESI):746.9[M+H] +

[0659] Step 13 4-((1R,3S,5s,7s)-5-aminoadamantan-2-yl)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide hydrochloride 25

[0660] 25n (30 mg, 0.04 mmol), 5 ml methanol and 30 mg Pd / C were added to the reaction flask, and then reacted in a hydrogen atmosphere for 2 hours. TLC showed that the raw materials had reacted completely. The reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure and diluted with a small amount of ethyl acetate. A solution of hydrogen chloride in ethyl acetate (2 M) was added, and solids precipitated. After filtration, pure 25 (10 mg, 0.015 mmol, 38%) was obtained.

[0661] MS m / z(ESI):612[M+H] +

[0662] 1 H NMR(400MHz, Methanol-d4)δ8.17(d,J=8.3Hz,1H),7.76–7.69(m,1H),7.53–7.45(m,2H),7.42(s ,1H),7.35(dd,J=8.7,1.5Hz,1H),6.83(h,J=4.1Hz,2H),6.74(tt,J=9.2,2.4Hz,1H),4.10(s,2H ),4.02-3.95(m,2H),3.90-3.78(m,1H),3.50-3.39(m,2H),3.20-3.11(m,1H),2.92-2.83(m,2H) ,2.25-2.08(m,4H),2.04-1.90(m,4H),1.88-1.69(m,4H),1.68-1.55(m,2H),1.35-1.26(m,1H).

[0663] Embodiment 26

[0664] Preparation of 4-(5-amino-adamantan-2-yl)-N-(5-(3,5-difluorobenzyl)-1H-indol-3-yl)-2-(piperidin-4-ylamino)benzamide 26

[0665]

[0666]

[0667] Step 1 Preparation of ethyl 4-((5-benzyloxycarbonylamino)adamantane)-2-yl)-2-((N-benzyloxycarbonylpiperidin-4-yl)amino)-benzoate 26a

[0668] 25i (310 mg, 0.69 mmol) and 4-Cbz-piperidone (cas: 19099-93-5, 806 mg, 3.46 mmol) were mixed and dissolved in 10 mL of acetic acid, and then sodium acetate borohydride (907 mg, 3.46 mmol) was added. Stir at room temperature overnight and the reaction was monitored by LCMS. The solvent was dried and the residue was separated by column and eluted with petroleum ether / ethyl acetate = 5 / 1 (V / V) to obtain the target compound 26a (340 mg) as a white solid in a yield of 74%.

[0669] LCMS: m / z=666.5 [M+H]+ Step 3 Preparation of 4-((5-benzyloxycarbonylamino)adamantane)-2-yl)-2-((N-benzyloxycarbonylpiperidin-4-yl)amino)-benzoic acid 26b

[0670] 26a (900 mg, 1.35 mmol) was dissolved in 15 mL of ethanol, and 8 mL of 4 M sodium hydroxide solution was added. The system was refluxed for 16 hours. The reaction was monitored by LCMS. After the reaction was completed, most of the ethanol in the solvent was removed by vortexing, and 10 mL of water was added to adjust the pH to 4, and solid precipitated. The solid was filtered out and washed with water to obtain the target compound 26b (780 mg) as a gray solid with a yield of 91%. It was used directly in the next step without further purification.

[0671] LCMS: m / z = 638.2 [M+H]+

[0672] Step 4 Preparation of 4-((5-benzyloxycarbonylamino)adamantane)-2-yl)-2-(N-(1-benzyloxycarbonylpiperidin-4-yl)-2,2,2-trifluoroacetylamino)-benzoic acid 26c

[0673] 26b (780 mg, 1.22 mmol) was dissolved in 20 mL of dichloromethane, and trifluoroacetic anhydride (1.2 eq) and triethylamine (1.5 eq) were added at 0°C. After the addition, the mixture was stirred at room temperature for three hours. The solvent was dried by spin drying to obtain the crude product 26c. It was used directly in the next step without purification.

[0674] LCMS: m / z=734.2[M+H]+

[0675] Step 5 Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((5-benzyloxycarbonylamino)adamantane)-2-yl)-2-(N-(1-benzyloxycarbonylpiperidin-4-yl)-2,2,2-trifluoroacetylamino)benzamide 26e

[0676] The crude product 26c (~1.22mmol) was dissolved in 20mL of dichloromethane, and 1mL of oxalyl chloride and 1 drop of DMF were added. The mixture was stirred at room temperature for 3 hours. After the reaction was completed, the solvent and excess oxalyl chloride were dried to obtain the acyl chloride 26d, which was dissolved in 10mL of anhydrous tetrahydrofuran for later use. 5-(3,5-difluorobenzyl)-1H-indazole-3-amine I-2 (316mg, 1.22mmol) was dissolved in 10mL of anhydrous tetrahydrofuran, and the above acyl chloride solution was added dropwise under ice-water cooling, and then triethylamine (0.33mL, 2.44mmol) was added. After the addition was completed, the mixture was stirred at room temperature overnight. The precipitated solid was filtered out, the filtrate was dried, and the residue was separated by column separation. The target compound 26e (410mg) was obtained as a yellow solid with a combined yield of 34% in the three steps.

[0677] LCMS: m / z = 975.2 [M+H] +

[0678] Step 6 Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((5-benzyloxycarbonylamino)adamantane)-2-yl)-2-((benzyloxycarbonyl)piperidin-4-ylamino)benzamide 26f

[0679] Add 26e (410 mg, 0.42 mmol) to a 20 mL sealed tube, dissolve in 5 mL methanol, and then add 1.12 mL triethylamine (20 eq). Seal the tube, heat and stir at 60°C for 1 hour. Monitor the reaction by LCMS. Spin dry the reaction system, and perform TLC separation using dichloromethane / methanol = 20 / 1 (V / V). Obtain the target compound 26f (240 mg) with a yield of 65%.

[0680] LCMS: m / z = 879.2 [M+H] +

[0681] Step 7 Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((5-amino)adamantanyl)-2-yl)-2-(piperidin-4-ylamino)benzamide 26

[0682] 26f (110 mg, 0.13 mmol) was added to a 50 mL single-mouth bottle, dissolved in 20 mL methanol, and 50 mg wet Pd / C (10%) was added. The air was replaced with a hydrogen balloon, and the mixture was stirred at room temperature for 20 hours under a hydrogen atmosphere. After the reaction was completed, Pd / C was filtered off, and the filtrate was dried to obtain the target compound 26 (40 mg) as a white solid with a yield of 50%.

[0683] LCMS: m / z = 611.2 [M+H]+

[0684] 1 HNMR(400MHz,MeOD)δ=8.03–8.05(d,J=8.0Hz,1H),7.88(s,1H),7.44–7.53(m,2 H),7.16–7.22(m,2H),6.73–6.85(m,3H),4.11(s,1H),3.97(brs,1H),3.57–3.5 9(d,J=6.8Hz,1H),3.42–3.46(m,1H),3.11–3.19(m,3H),2.83(s,2H),2.25–2.2 9(m,2H),2.12(s,4H),1.82–1.99(m,6H),1.57–1.62(m,2H),1.14–1.26(m,1H).

[0685] Embodiment 27

[0686] Preparation of (R)-N-(5-(2-(2,5-difluorophenyl)pyrrolidin-1-yl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 27

[0687]

[0688] Using the same preparation method as in Example 3 and using (R)-2-(2,5-difluorophenyl)pyrrolidine as a raw material, Compound 27 can be obtained.

[0689] MS m / z(ESI):616[M+1] +

[0690] 1H NMR (400MHz, DMSO-d6) δ12.32(s,1H),9.94(s,1H),8.28(d,J=7.7Hz,1H),7.76(d,J=9.0Hz,1H),7.3 1–7.19(m,2H),7.14–7.04(m,1H),6.87(m,1H),6.66(dd,J=9.1,2.3Hz,1H),6.49(d,J=2.2Hz,1H),6 .22(dd,J=9.0,2.3Hz,1H),6.14(d,J=2.3Hz,1H),4.91(d,J=8.3Hz,1H),3.83(m,2H),3.71(m,2H),3 .56–3.45(m,2H),3.27(m,5H),2.48–2.36(m,4H),2.24(s,3H),2.05–1.80(m,5H),1.42–1.25(m,3H).

[0691] Embodiment 28

[0692] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 28

[0693]

[0694] By adopting the same preparation method as Example 10 and using 2,5-difluorobromobenzene as raw material, Compound 28 can be obtained.

[0695] MS m / z(ESI):591[M+H] + .

[0696] 1H NMR (400MHz, DMSO-d6) δ12.61(s,1H),10.07(s,1H),8.28(d,J=7.7Hz,1H),7.75(d,J=9.1Hz,1H),7.42(s,1H),7.35(d, J=8.6Hz,1H),7.32–7.21(m,2H),7.13(td,J=9.3,4.6Hz,1H),7.04(td,J=8.7,8.2,4.2Hz,1H),6.20(dd,J=9.0,2.3Hz,1 H),6.10(d,J=2.4Hz,1H),4.90(t,J=5.2Hz,1H),4.38(t,J=7.4Hz,1H),4.03–3.84(m,2H),3.78(d,J=11.8Hz,2H),3.71 –3.58(m,1H),3.46(t,J=10.9Hz,2H),3.23(s,4H),2.43(s,4H),2.21(s,3H),1.90(d,J=12.7Hz,2H),1.41–1.25(m,2H).

[0697] Embodiment 29

[0698] Preparation of ethyl 2-(3,5-difluorophenyl)-2-(3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamido)-1H-indazol-5-yl)acetate 29

[0699]

[0700]

[0701] Step 1 5-(2-(Benzyloxy)-1-(3,5-difluorophenyl)ethyl)-3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4)tert-butyl)amino)benzamido)-1H-indazole-1-carboxylic acid tert-butyl ester 29a

[0702] A solution of compound 10g (1g, 1.47mmol) in DCM (50mL) was added dropwise to Boc2O (320mg, 56.8mmol) at 0°C, and the reaction solution was stirred at room temperature for 2 hours. Then, ice water (150mL) was carefully added to the reaction solution, and the reaction solution was continued to be stirred at room temperature for 3 hours. The reaction solution was concentrated and extracted with DCM (200mL). The organic phase was washed with saturated sodium bicarbonate solution and brine, respectively, and the crude product obtained after drying and spin drying was purified by column chromatography to obtain a crude product of compound 29a (1.0g, 1.28mmol).

[0703] MS m / z(ESI):781[M+H] + .

[0704] Step 2: tert-Butyl 5-(1-(3,5-difluorophenyl)-2-hydroxyethyl)-3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl))tert-butylamino)benzamido)-1H-indazole-1-carboxylate 29b

[0705] Compound 29a (1.0 g, 1.28 mmol) was dissolved in methanol (60 mL), and ammonium formate (400 mg, 6.4 mmol) and palladium carbon (100 mg) were added, and the reaction solution was heated to 90°C and refluxed for 2 h. The initial product was filtered and dried, and then purified by silica gel column chromatography to obtain compound 29b (500 mg, 0.72 mmol).

[0706] MS m / z(ESI):691[M+H] + .

[0707] Step 3 5-(2-acetoxy-1-(3,5-difluorophenyl)ethyl)-3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)tert-butyl)amino)benzamido)-1H-indazole-1-carboxylic acid tert-butyl ester 29c

[0708] Compound 29b (100 mg, 0.14 mmol) was dissolved in DCM (15 mL), cooled to 0°C, acetic anhydride (14 mg, 0.14 mmol) was added, and the reaction solution was stirred at 0°C for 1 hour. Water was then added to the reaction solution, extracted with DCM (50 mL), and the organic phase was washed with saturated brine, dried and spin-dried to give a crude product, which was purified by column chromatography to give compound 036c (60 mg, 0.082 mmol).

[0709] MS m / z(ESI):733[M+H] + .

[0710] Step 4: Ethyl 2-(3,5-difluorophenyl)-2-(3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamido)-1H-indazol-5-yl)acetate 29

[0711] Compound 29c (60 mg, 0.082 mmol) was dissolved in DCM (10 mL), and TFA (2 mL) was added. The mixture was stirred at room temperature for 1 h, and then concentrated directly. After purification by column chromatography, compound 29 trifluoroacetate (40 mg, 0.063 mmol) was obtained.

[0712] MS m / z(ESI):633[M+H]+ .

[0713] 1 H NMR (400MHz, DMSO-d6) δ12.70(s,1H),10.20(s,1H),8.32(dd,J=4.0,2.4Hz,1H),7.81(d,J=8.8Hz,1H),7.50 (s,1H),7.41(d,J=8.9Hz,1H),7.37–7.31(m,1H),7.24(d,J=7.2Hz,1H),7.10–7.01(m,2H),6.26(d,J=8.7Hz ,1H),6.17(s,1H),4.72(s,1H),4.65–4.59(m,1H),4.07–3.95(m,2H),3.84–3.75(m,2H),3.73-3.68(m,1H), 3.60–3.56(m,4H),3.50–3.45(m,2H),3.12–3.09(m,4H),2.82(s,3H),1.99–1.81(m,5H),1.37–1.29(m,2H).

[0714] Embodiment 30

[0715] Preparation of N-(5-(1-(3,5-difluorophenyl)ethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 30

[0716]

[0717] Using the same preparation method as Example 17 and using 1-(3,5-difluorophenyl)-1-bromoethane as raw material, Compound 30 can be obtained.

[0718] MS m / z(ESI):576[M+1] +

[0719] 1H NMR(400MHz,Chloroform-d)δ8.85(s,1H),8.63(s,1H),8.13(d,J=7.3Hz,1H),7.78(s,1H),7. 53(d,J=8.9Hz,1H),6.88–6.77(m,2H),6.58(tt,J=8.8,2.3Hz,1H),6.22(dd,J=9.0,2.2Hz,1H ),6.12–6.05(m,1H),4.37(q,J=7.1Hz,1H),3.99(d,J=11.7Hz,2H),3.66–3.45(m,3H),3.35(t ,J=5.2Hz,4H),2.63(t,J=5.0Hz,4H),2.40(s,3H),2.03(d,J=13.2Hz,2H),1.75–1.56(m,5H).

[0720] Embodiment 31

[0721] Preparation of N-(5-(3-fluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 31

[0722]

[0723] By adopting the same preparation method as Example 17 and using 3-fluorobenzyl bromide as the starting material, Compound 31 can be obtained.

[0724] MS m / z(ESI):544[M+1] +

[0725] 1H NMR (400MHz, DMSO-d6) δ13.17(s,1H),10.30(s,1H),8.90(s,1H),8.24(d,J=7.6Hz,1H),7.79(d,J=9.1Hz,1H),7 .50(s,1H),7.30(td,J=8.0,6.3Hz,1H),7.16–7.06(m,2H),6.99(td,J=8.7,2.8Hz,1H),6.24(dd,J=9.0,2.3Hz, 1H),6.13(d,J=2.4Hz,1H),4.18(s,2H),3.82(dt,J=11.7,4.0Hz,2H),3.67(t,J=8.9Hz,1H),3.54–3.43(m,2H), 3.28(t,J=5.0Hz,4H),2.46(d,J=5.2Hz,4H),2.24(s,3H),1.97–1.89(m,2H),1.36(ddd,J=13.4,9.9,3.8Hz,2H).

[0726] Embodiment 32

[0727] Preparation of N-(5-(1-(3-fluorophenyl)-2-hydroxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 32

[0728]

[0729] By adopting the same preparation method as Example 10 and using 3-fluorobromobenzene as raw material, compound 32 can be obtained.

[0730] MS m / z(ESI):573[M+H] + .

[0731] 1H NMR (400MHz, DMSO-d6) δ12.59(s,1H),10.05(s,1H),8.28(d,J=7.7Hz,1H),7.76(d,J=9.1Hz,1H),7.45(d,J=1.4Hz,1H),7.35(d,J= 8.6Hz,1H),7.31–7.19(m,2H),7.15–7.03(m,2H),7.00–6.89(m,1H),6.21(dd,J=9.0,2.3Hz,1H),6.11(d,J=2.3Hz,1H),4.81(t,J=5 .1Hz,1H),4.16(t,J=7.3Hz,1H),3.94(td,J=7.7,7.2,4.8Hz,2H),3.78(dt,J=11.6,4.1Hz,2H),3.66(dq,J=13.3,5.0Hz,1H),3.46 (td,J=11.0,2.5Hz,2H),3.24(d,J=5.5Hz,4H),2.47(d,J=3.1Hz,4H),2.23(s,3H),1.98–1.82(m,2H),1.32(dt,J=12.8,3.2Hz,2H).

[0732] Embodiment 33

[0733] Preparation of N-(5-(1-(3-fluorophenyl)ethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 33

[0734]

[0735] By adopting the same preparation method as Example 17, using 3,4-dihydro-2H-pyran and 1-(3-fluorophenyl)-1-bromoethane as raw materials, Compound 33 can be obtained.

[0736] MS m / z(ESI):558[M+1] +

[0737] 1H NMR(400MHz,Chloroform-d)δ8.87–8.81(m,1H),8.62(s,1H),8.12(d,J=16.7Hz,1H),7.76(d,J=1.2Hz,1H),7.53(d,J=8.9H z,1H),7.21(td,J=8.0,6.0Hz,1H),7.11–7.06(m,1H),7.02(dt,J=10.3,2.2Hz,1H),6.83(tdd,J=8.3,2.6,1.1Hz,1H),6.21( dd,J=9.1,2.2Hz,1H),6.09(d,J=2.3Hz,1H),4.41(q,J=7.1Hz,1H),3.99(d,J=11.7Hz,2H),3.58(q,J=13.0,10.8Hz,3H),3.3 5(t,J=5.1Hz,4H),2.66(t,J=5.0Hz,4H),2.42(s,3H),2.03(d,J=14.5Hz,2H),1.73(d,J=7.2Hz,3H),1.62(d,J=11.9Hz,2H).

[0738] Embodiment 34

[0739] Preparation of N-(5-(1-(2,5-difluorophenyl)ethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 34

[0740]

[0741] By adopting the same preparation method as Example 33, using 3,4-dihydro-2H-pyran and 1-(3-fluorophenyl)-1-bromoethane as raw materials, Compound 34 can be obtained.

[0742] MS m / z(ESI):576[M+1] +

[0743] 1H NMR(400MHz,Chloroform-d)δ8.85(s,1H),8.64(s,1H),8.17(d,J=7.3Hz,1H),7.79(s,1H),7.53(d,J= 8.9Hz,1H),7.04(ddd,J=9.2,5.8,3.2Hz,1H),6.91(td,J=9.1,4.6Hz,1H),6.80(ddd,J=8.4,6.2,3.5Hz ,1H),6.22–6.16(m,1H),6.08(d,J=2.3Hz,1H),4.70(q,J=7.2Hz,1H),3.98(d,J=11.7Hz,2H),3.67–3. 50(m,3H),3.43–3.30(m,4H),2.67(t,J=5.0Hz,4H),2.43(s,3H),2.07–1.98(m,2H),1.76–1.56(m,5H).

[0744] Embodiment 35

[0745] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-methoxyethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 35

[0746]

[0747] Step 1 5-Bromo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridine 35b

[0748] Compound 35a (10 g, 50.5 mol), DHP (7.6 g, 90.91 mmol), methanesulfonic acid (484 mg, 5.05 mmol) were dissolved in dichloromethane (200 mL) and stirred overnight at room temperature to complete the reaction. Saturated sodium bicarbonate was added and extracted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography to obtain the title compound 2 (7 g, 24.822 mmol).

[0749] MS m / z(ESI):282.1[M+1] +

[0750] Step 2 1-(Tetrahydro-2H-pyran-2-yl)-5-vinyl-1H-pyrazolo[3,4-c]pyridine 35c

[0751] Compound 35b (5.62 g, 20 mmol) and 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (6.16 g, 40 mmol) were dissolved in 1,4-dioxane (200 mL) and water (40 mL), and Pd(PPh3)4 (2.308 g, 2 mmol) and sodium carbonate (4.24 g, 40 mmol) were added. The mixture was heated at 100°C and stirred for 6 h under the protection of argon gas, and the reaction was completed. The reaction solution was added with water and extracted with ethyl acetate three times, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the title compound 35c (3.4 g, 14.847 mmol).

[0752] MS m / z(ESI):230.1[M+H] +

[0753] Step 3 5-(1,2-dibromoethyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridine 35d

[0754] Compound 35c (3.4 g, 14.847 mmol) was dissolved in acetic acid (30 mL), lithium bromide (1.547 g, 17.816 mmol) was added, and the mixture was stirred at room temperature for five minutes, and then sodium periodate (794 mg, 3.712 mmol) was added. After stirring at room temperature for 4 hours, the reaction was completed. Water and ethyl acetate were added to the reaction solution for extraction, and the organic phase was washed with saturated sodium carbonate and brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the title compound 35d (4 g, 10.28 mmol), with a yield of 69%.

[0755] MS m / z(ESI)390.1[M+1] +

[0756] Step 4: 5-(1-bromovinyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridine 35e

[0757] Compound 35d (4 g, 10.28 mmol) was dissolved in methanol (20 mL) and tetrahydrofuran (20 mL), and potassium carbonate (2.838 g, 20.56 mmol) was added. After stirring overnight at room temperature, the reaction was completed. The reaction solution was spin-dried and water was added, extracted three times with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the title compound 35e (1.4 g, 4.545 mmol).

[0758] MS m / z(ESI):310.1[M+H] +

[0759] Step 5 5-(1-(2,5-difluorophenyl)vinyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridine 35f

[0760] Compound 35e (1.4 g, 4.54 mmol) and (2,5-difluorophenyl)boric acid (1.436 mg, 9.09 mmol) were dissolved in 1,4-dioxane (40 mL) and water (10 mL), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (322 mg, 0.454 mmol) and sodium carbonate (963 mg, 9.09 mmol) were added. The mixture was heated at 100°C and stirred for 5 h under the protection of argon gas, and the reaction was completed. Water and ethyl acetate were added to the reaction solution for extraction, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the title compound 35f (1 g, 2.932 mmol).

[0761] MS m / z(ESI):342.2[M+1] +

[0762] Step 6: (2-(2,5-difluorophenyl)-2-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridin-5-yl)ethyl)boronic acid 35 g

[0763] Compound 35f (720 mg, 2.11 mmol) and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (1072 mg, 4.22 mmol) were dissolved in methanol (30 mL), and cuprous oxide (151 mg, 1.055 mmol), KH2PO4 (918 mg, 5.2775 mmol), and triphenylphosphine (274 mg, 1.055 mmol) were added and heated under reflux for 4 h. The reaction was completed. After column chromatography purification, the title compound 35g (320 mg, 0.826 mmol) was obtained with a yield of 39%.

[0764] MS m / z(ESI):388.3[M+H] +

[0765] Step 7: 2-(2,5-difluorophenyl)-2-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridin-5-yl)ethan-1-ol 35h

[0766] Compound 35g (320 mg, 0.826 mmol) was dissolved in tetrahydrofuran (6 mL), and 1M sodium hydroxide aqueous solution (6 mL) was added. The reaction solution was cooled to 0°C, and 30% hydrogen peroxide (3 mL) was slowly added dropwise. The reaction solution was stirred at room temperature for 3 hours, and then quenched with saturated sodium thiosulfate solution. It was extracted with ethyl acetate (150 mL), and the organic phase was washed with saturated sodium bicarbonate solution and brine, dried with anhydrous sodium sulfate, filtered and dried to obtain the initial product, and purified by silica gel column chromatography to obtain compound 35h (260 mg, 0.724 mmol) with a yield of 87%.

[0767] MS m / z(ESI):360.2[M+H] +

[0768] Step 8 5-(1-(2,5-difluorophenyl)-2-methoxyethyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridine 35i

[0769] Compound 35h (260 mg, 0.724 mmol) was dissolved in tetrahydrofuran (10 mL), and sodium hydride (87 mg, 2.1727 mmol) was added under the protection of argon in an ice bath, and iodomethane (308 mg, 2.1727 mol) was added after stirring for half an hour. The temperature was slowly raised to room temperature and stirred overnight to complete the reaction. The reaction solution was quenched with dilute hydrochloric acid, saturated sodium bicarbonate was added, and the mixture was extracted three times with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the title compound 35i (210 mg, 0.7266 mmol).

[0770] MS m / z(ESI):374.2[M+H] +

[0771] Step 9 5-(1-(2,5-difluorophenyl)-2-methoxyethyl)-1H-pyrazolo[3,4-c]pyridine 35j

[0772] Compound 35i (260 mg, 0.697 mmol) was dissolved in dichloromethane (4 mL), trifluoroacetic acid (4 mL) was added, and the reaction was completed after 2 h at room temperature. After concentration, the title compound 35j (210 mg) was purified by plate chromatography, which was not pure and was directly used for the next step.

[0773] MS m / z(ESI):290.1[M+H] +

[0774] Step 10 5-(1-(2,5-difluorophenyl)-2-methoxyethyl)-3-iodo-1H-pyrazolo[3,4-c]pyridine 35k

[0775] Compound 35j (210 mg, 0.7266 mmol) was dissolved in DMF (5 ml), K2CO3 (401 mg, 2.905 mmol) and 9a (367.6 mg, 1.4532 mmol) were added in sequence, and the mixture was reacted at room temperature for 3 h. EA (50 ml) was added for dilution, and the mixture was washed with H2O (25 ml) and brine (15 ml×2) in sequence, dried over anhydrous sodium sulfate, concentrated, and purified by column to obtain the title compound 35k (250 mg, 0.6 mmol), with a yield of 83%.

[0776] MS m / z(ESI):416.1[M+1] +

[0777] Step 11 5-(1-(2,5-difluorophenyl)-2-methoxyethyl)-3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridine 35l

[0778] Compound 35k (250 mg, 0.602 mol) and DHP (101 mg, 1.204 mmol) were dissolved in dichloromethane (20 mL). Methanesulfonic acid (116 mg, 1.204 mmol) was added dropwise under ice bath. After stirring at room temperature overnight, the reaction was completed. Saturated sodium bicarbonate was added and then extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the title compound 35l (160 mg, 0.32 mmol). The yield was 53%.

[0779] MS m / z(ESI):500.2[M+1] +

[0780] Step 12 5-(1-(2,5-difluorophenyl)-2-methoxyethyl)-3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridine 35m

[0781] Compound 351 (80 mg, 0.16 mmol), 4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide (76 mg, 0.24 mmol), Cu (10 mg, 0.16 mmol), CuI (31 mg, 0.16 mmol), N,N'-dimethylethylenediamine (85 mg, 0.96 mmol,) and K3PO4 (102 mg, 0.48 mmol) were added to toluene (15 ml) and purged with argon for 5 times. The reaction mixture was refluxed for 6 h. The reaction solution was directly mixed and purified by column to obtain compound 35m (48 mg, 0.069 mmol) with a yield of 43.6%.

[0782] MS m / z(ESI):690.6[M+1]+

[0783] Step 13 N-(5-(1-(2,5-difluorophenyl)-2-methoxyethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide HHT13056

[0784] Compound 13 (48 mg, 0.069 mmol) was dissolved in dichloromethane (4 mL), trifluoroacetic acid (4 mL) was added, and the reaction was completed after 2 h at room temperature. After spin drying, saturated sodium bicarbonate was added and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by preparative TLC to obtain the title compound HHT13056 (31 mg, 0.0512 mmol), with a yield of 73%.

[0785] MS m / z(ESI):606.5[M+1] +

[0786] 1 H NMR (400MHz, DMSO-d6) δ13.17(s,1H),10.29(s,1H),8.90(d,J=1.2Hz,1H),8.23(d,J=7.7Hz,1H),7.75(d,J=9.1 Hz,1H),7.50(d,J=1.2Hz,1H),7.31(s,1H),7.15(dd,J=9.3,4.7Hz,1H),7.06(d,J=4.1Hz,1H),6.24–6.19(m,1H ),6.10(d,J=2.4Hz,1H),4.70(t,J=7.3Hz,1H),4.02–3.91(m,2H),3.83–3.75(m,2H),3.71–3.63(m,1H),3.47(t ,J=10.5Hz,2H),3.23(d,J=10.2Hz,7H),2.42(s,4H),2.20(s,3H),1.90(d,J=12.6Hz,2H),1.31(d,J=9.6Hz,2H).

[0787] Embodiment 36

[0788] Preparation of 2-(3-fluorophenyl)-2-(3-(4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)-1H-indazol-5-yl)ethyl isobutyl ester 36

[0789]

[0790] Using the same preparation method as Example 29, with 3-fluorobromobenzene and isobutyric anhydride as raw materials, Compound 36 can be obtained.

[0791] MS m / z(ESI):643[M+1] + .

[0792] 1 H NMR (400MHz, DMSO-d6) δ12.62(s,1H),10.06(s,1H),8.26(d,J=7.7Hz,1H),7.75(d,J=9.1Hz,1H),7.52(s,1H),7.42–7.2 3(m,3H),7.20–7.08(m,2H),6.99(td,J=8.7,2.2Hz,1H),6.20(dd,J=9.0,2.3Hz,1H),6.10(d,J=2.3Hz,1H),4.58(qd,J=1 0.9,7.5Hz,2H),4.46(t,J=7.5Hz,1H),3.78(d,J=11.6Hz,2H),3.71–3.60(m,1H),3.46(t,J=10.9Hz,2H),3.23(t,J=5.0H z,4H),2.44–2.30(m,5H),2.20(s,3H),1.90(d,J=13.0Hz,2H),1.31(q,J=12.0,10.2Hz,2H),0.88(dd,J=7.0,2.0Hz,6H).

[0793] Embodiment 37

[0794] Preparation of N-(5-(3-fluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(5-methyl-2,5-diazabicyclo[2.2.1]hept-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 37

[0795]

[0796] By adopting the same preparation method as Example 17, using 3-fluorobromobenzene and 2-methyl-2,5-diazabicyclo[2.2.1]heptane as raw materials, Compound 37 can be obtained.

[0797] MS m / z(ESI):556[M+1] +

[0798] 1H NMR (400MHz, DMSO-d6) δ13.15(s,1H),10.21(s,1H),8.89(d,J=1.3Hz,1H),8.31(d,J=7.6Hz,1H),7.76(d,J=9.0 Hz,1H),7.49(d,J=1.3Hz,1H),7.30(td,J=7.9,6.3Hz,1H),7.16–7.07(m,2H),6.99(td,J=8.3,2.5Hz,1H),5.94( dd,J=8.8,2.2Hz,1H),5.77(d,J=2.2Hz,1H),4.50(s,1H),4.18(s,2H),3.88–3.76(m,2H),3.66(s,2H),3.55–3.4 4(m,3H),3.35-3.28(m,2H),2.91(d,J=9.8Hz,1H),2.43(s,3H),1.92(q,J=14.2,13.1Hz,4H),1.42–1.29(m,2H).

[0799] Embodiment 38

[0800] Preparation of N-(5-(1-(3-fluorophenyl)-2-methoxyethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 38

[0801]

[0802] Using the same preparation method as Example 23 and using 3-fluorobromobenzene as raw material, Compound 38 can be obtained.

[0803] MS m / z(ESI):587[M+1] + .

[0804] 1H NMR (400MHz, DMSO-d6) δ12.59(s,1H),10.05(s,1H),8.30(d,J=7.7Hz,1H),7.76(d,J=9.1Hz,1H),7.47(s ,1H),7.35(d,J=8.7Hz,1H),7.30–7.23(m,2H),7.18–7.03(m,2H),7.00–6.88(m,1H),6.21(dd,J=9.0,2. 3Hz,1H),6.10(d,J=2.3Hz,1H),4.36(t,J=7.4Hz,1H),3.97–3.72(m,4H),3.72–3.59(m,1H),3.52–3.42( m,2H),3.23(d,J=10.0Hz,7H),2.48(s,4H),2.22(s,3H),1.90(dt,J=13.0,4.2Hz,2H),1.42–1.24(m,2H).

[0805] Embodiment 39

[0806] Synthesis of N-(5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-(5-methyl-2,5-diazabicyclo[2.2.1]hept-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 39

[0807]

[0808] Using the same preparation method as Example 17 and using 2-methyl-2,5-diazabicyclo[2.2.1]heptane as raw material, Compound 39 can be obtained.

[0809] MS m / z(ESI):574[M+1] +

[0810] 1H NMR (400MHz, DMSO-d6) δ13.14(s,1H),10.19(s,1H),8.90(d,J=1.2Hz,1H),8.31(d,J=7.5Hz,1H ),7.75(d,J=9.0Hz,1H),7.53(s,1H),7.07–6.95(m,3H),5.93(d,J=8.7Hz,1H),5.76(s,1H),4.4 2(s,1H),4.19(s,2H),3.82(d,J=11.4Hz,2H),3.64(s,1H),3.48(t,J=10.8Hz,4H),3.32-3.28( m,2H),2.83(d,J=9.3Hz,1H),2.30(s,3H),1.92(q,J=12.5,12.0Hz,4H),1.35(d,J=11.5Hz,2H).

[0811] Embodiment 40

[0812] Synthesis of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(3-(dimethylamino)azetidin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 40

[0813]

[0814] By adopting the same preparation method as Example 12 and using N,N-dimethylazacyclo-3-amine as raw material, compound 40 can be obtained.

[0815] MS m / z(ESI):561[M+1] +

[0816] 1H NMR (400MHz, DMSO-d6) δ12.64(s,1H),10.05(s,1H),8.38(d,J=7.5Hz,1H),7.78(d,J=8.8Hz,1H),7.48(s,1 H),7.40(d,J=8.5Hz,1H),7.25(dd,J=8.6,1.6Hz,1H),7.06–6.92(m,3H),5.78–5.68(m,1H),5.63(d,J=2.2 Hz,1H),4.03(s,2H),3.97(t,J=7.4Hz,2H),3.81(dt,J=11.6,3.9Hz,2H),3.72–3.53(m,3H),3.47(td,J=11 .5,10.9,2.4Hz,2H),3.27-3.19(m,1H),2.17(s,6H),1.98–1.87(m,2H),1.34(tdd,J=13.4,8.6,3.6Hz,2H).

[0817] Embodiment 41

[0818] Synthesis of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(3-((2-methoxyethyl)(methyl)amino)azetidin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 41

[0819]

[0820] By adopting the same preparation method as Example 12 and using N-(2-methoxyethyl)-N-methylazepine-3-amine as raw material, Compound 41 can be obtained.

[0821] MS m / z(ESI):605[M+1] +

[0822] 1H NMR (400MHz, DMSO-d6) δ12.64(s,1H),10.04(s,1H),8.38(d,J=7.6Hz,1H),7.78(d,J=8.8Hz,1H),7.48(s,1 H),7.43–7.35(m,1H),7.25(dd,J=8.6,1.6Hz,1H),7.01(ddd,J=11.8,8.0,1.5Hz,3H),5.72(dd,J=8.7,2.1 Hz,1H),5.63(d,J=2.1Hz,1H),4.03(s,2H),3.96(t,J=7.5Hz,2H),3.80(dd,J=9.8,5.9Hz,2H),3.66-3.55( m,3H),3.52–3.37(m,5H),3.24(s,3H),2.46(t,2H),2.17(s,3H),1.93(d,J=12.8Hz,2H),1.38-1.28(m,2H).

[0823] Embodiment 42

[0824] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 42

[0825]

[0826] Using the same preparation method as Example 12 and using 2-(pyrrolidin-1-yl)ethylamine as raw material, Compound 42 can be obtained.

[0827] MS m / z(ESI):575[M+H]+.

[0828] 1 H NMR (400MHz, Methanol-d4) δ7.68(d,J=8.8Hz,1H),7.56(s,1H),7.40(d,J=8.6Hz,1H),7.25–7.19(m,1H),6.85–6.76(m,2H),6.75–6.66(m,1 H),6.07–5.95(m,2H),4.05(s,2H),3.91(d,J=11.7Hz,2H),3.69–3.44(m,6H),3.15(d,J=13.6Hz,5H),2.07–1.92(m,6H),1.54–1.45(m,2H).

[0829] Embodiment 43

[0830] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 43

[0831]

[0832]

[0833] Step 1 Synthesis of tert-butyl 4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-nitrobenzoate 43a

[0834] I-1b (500 mg, 2.1 mmol), 2N-ethyl-2-(pyrrolidin-1-yl)ethane-1-amine (587 mg, 4.1 mmol) and DIPEA (1060 mg, 8.3 mmol) were dissolved in NMP and stirred at 120°C for 2 hours. The mixture was cooled to room temperature, quenched with water, extracted with ethyl acetate, and the organic phases were combined and washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified by column chromatography (DCM: MeOH = 15:1) to obtain compound 43a (500 mg) as a yellow oil.

[0835] MS m / z(ESI):364(M+H) +

[0836] Step 2 Synthesis of tert-butyl 2-amino-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)benzoate 43b

[0837] 43a (500 mg, 1.37 mmol) was dissolved in methanol (14 mL), and 5 mL of saturated aqueous ammonium chloride solution and zinc powder (447 mg, 6.88 mmol) were added. The mixture was stirred at room temperature for 2 hours, filtered, and the filtrate was extracted with DCM. The organic phase was washed with water, dried over anhydrous sodium sulfate, and concentrated to give a crude product 43b (360 mg) as a white solid. The crude product was used directly in the next step.

[0838] MS m / z(ESI):334(M+H) +

[0839] Step 3 Synthesis of tert-butyl 4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzoate 43c

[0840] Tetramethylammonium triacetoxyborohydride (1130 mg, 4.32 mmol) was slowly added to a solution of compound 43b (360 mg, 1.08 mmol) in DCM (10 mL) and trifluoroacetic acid (1 mL), and the mixture was stirred at room temperature for 2 hours. Saturated aqueous NaHCO3 solution was added to quench the reaction. The system was extracted with DCM, the organic phase was washed with water, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by column chromatography (DCM: MeOH = 10: 1) to give compound 43c (300 mg) as a yellow solid in a yield of 66%.

[0841] MS m / z(ESI):418(M+H) +

[0842] Step 4 Synthesis of tert-butyl 4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoate 43d

[0843] Compound 43c (300 mg, 0.71 mmol) and triethylamine (286 mg, 2.84 mmol) were dispersed in DCM, trifluoroacetic anhydride (302 mg, 1.43 mmol) was added dropwise at room temperature, and the mixture was stirred at room temperature for 2 hours. Water was added to quench the reaction. The system was extracted with DCM, the organic phase was washed with water, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by column chromatography (DCM: MeOH = 15: 1) to obtain compound 43d (250 mg) as a yellow oil with a yield of 68%.

[0844] MS m / z(ESI):514(M+H) +

[0845] Step 5 Synthesis of 4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoic acid 43e

[0846] Compound 43d (250 mg, 0.48 mmol) was dispersed in DCM (5 mL), trifluoroacetic acid (5 mL) was added dropwise at room temperature, the mixture was stirred at room temperature for 2 hours, concentrated, and the crude compound 43e was directly used in the next reaction.

[0847] MS m / z(ESI):458(M+H) +

[0848] Step 6 Synthesis of 4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzoyl chloride 43f

[0849] Compound 43e (200 mg, 0.43 mmol) was dispersed in DCM (5 mL), and oxalyl chloride (273 mg, 2.15 mmol) and a drop of DMF were added dropwise at room temperature. The mixture was stirred at room temperature for 2 hours and concentrated. The crude compound 43f was directly used in the next reaction.

[0850] Step 7: Synthesis of 43 g of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide

[0851] Compound I-2 (98 mg, 0.37 mmol) and triethylamine (143 mg, 1.41 mmol) were dissolved in DCM, and a DCM solution of 43f (180 mg, 0.37) was added dropwise under an ice bath. After the addition was complete, the mixture was warmed to room temperature and stirred for 2 hours. Water was added to quench the mixture, and the mixture was extracted with DCM. The organic phase was washed with water, dried over anhydrous sodium sulfate, and concentrated. The crude compound 43g was directly used in the next reaction.

[0852] MS m / z(ESI):699(M+H) +

[0853] Step 8 Synthesis of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 43

[0854] Compound 43g (80 mg, 0.11 mmol) and triethylamine (151 mg, 1.5 mmol) were dissolved in methanol (2 mL), stirred at 60°C for 2 hours, concentrated, and the crude product was purified by column chromatography (DCM:MeOH=10:1) to give compound 43 (16.5 mg) as a white solid.

[0855] MS m / z(ESI):603(M+H) +

[0856] 1H NMR (400MHz, Methanol-d4) δ7.70(d,J=9.0Hz,1H),7.57(dd,J=1.7,0.8Hz,1H),7.40(dd,J=8.6,0.8Hz,1H),7.22 (dd,J=8.7,1.6Hz,1H),6.85–6.77(m,2H),6.70(tt,J=9.0,2.3Hz,1H),6.12(dd,J=9.0,2.5Hz,1H),5.95(d,J=2. 4Hz,1H),4.05(s,2H),3.92(dt,J=11.8,4.1Hz,2H),3.66(dt,J=9.4,5.1Hz,1H),3.60–3.50(m,4H),3.45(q,J=7. 1Hz,2H),2.80–2.64(m,6H),2.03(d,J=13.2Hz,2H),1.89–1.81(m,4H),1.59–1.46(m,2H),1.19(t,J=7.0Hz,3H).

[0857] Embodiment 44

[0858] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(dimethylamino)ethyl)thio)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 44

[0859]

[0860] Using the same preparation method as Example 43 and using 2-(dimethylamino)ethanethiol as raw material, Compound 44 can be obtained.

[0861] MS m / z(ESI):562(M+H) +

[0862] 1H NMR (400MHz, DMSO-d6) δ12.68(s,1H),10.37(s,1H),8.08(d,J=7.6Hz,1H),7.79(d,J=8.5Hz,1H),7. 46(s,1H),7.38(d,J=8.5Hz,1H),7.23(dd,J=8.6,1.5Hz,1H),7.01–6.90(m,3H),6.61(d,J=1.7Hz,1 H),6.48(dd,J=8.4,1.7Hz,1H),4.01(s,2H),3.84–3.73(m,2H),3.64(d,J=7.5Hz,1H),3.48–3.38(m ,2H),3.15–3.05(m,2H),2.54–2.49(m,2H),2.17(s,6H),1.89(d,J=12.5Hz,2H),1.41–1.26(m,2H).

[0863] Embodiment 45

[0864] Preparation of N-(5-(3-fluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 45

[0865]

[0866] Using the same preparation method as Example 43 and using 3-fluorobromobenzene as raw material, Compound 45 can be obtained.

[0867] LCMS (ESI-MS) m / z: 585 (M+H + ).

[0868] 1H NMR (400MHz, Methanol-d4) δ7.70(d,J=9.0Hz,1H),7.55(dd,J=1.7,0.9Hz,1H),7.38(dd,J=8.7,0.9Hz,1H),7.28–7 .19(m,2H),7.03(d,J=7.6Hz,1H),6.96–6.82(m,2H),6.12(dd,J=9.0,2.4Hz,1H),5.94(d,J=2.4Hz,1H),4.05(s,2H ),3.92(dt,J=11.9,4.1Hz,2H),3.67(dt,J=9.4,5.1Hz,1H),3.61–3.50(m,4H),3.45(q,J=7.0Hz,2H),2.83–2.69(m ,6H),2.03(d,J=13.1Hz,2H),1.87(q,J=4.9,3.7Hz,4H),1.52(ddt,J=17.7,9.8,4.1Hz,2H),1.19(t,J=7.0Hz,3H).

[0869] Embodiment 46

[0870] Preparation of N-(5-(3-fluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 46

[0871]

[0872] By adopting the same preparation method as Example 45 and using N-methyl-2-(pyrrolidin-1-yl)ethylamine as raw material, Compound 46 can be obtained.

[0873] LCMS (ESI-MS) m / z: 571 (M+H + ).

[0874] 1H NMR (400MHz, Methanol-d4) δ7.71(d,J=9.0Hz,1H),7.55(q,J=0.9Hz,1H),7.38(dd,J=8.7,0.9Hz,1 H),7.27–7.20(m,2H),7.03(d,J=7.8Hz,1H),6.95–6.83(m,2H),6.14(dd,J=9.1,2.4Hz,1H),5.97(d ,J=2.4Hz,1H),4.05(s,2H),3.92(dt,J=11.9,4.0Hz,2H),3.68(dt,J=9.5,5.2Hz,1H),3.62–3.53(m ,4H),3.03(s,3H),2.84–2.71(m,6H),2.03(d,J=13.8Hz,2H),1.89–1.84(m,4H),1.57–1.47(m,2H).

[0875] Embodiment 47

[0876] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((oxetan-3-ylmethyl)amino)benzamide 47

[0877]

[0878] By adopting the same preparation method as Example 11 and using oxetane-3-ylcarboxaldehyde as raw material, compound 47 can be obtained.

[0879] LCMS (ESI-MS) m / z: 547.4 (M+H + ).

[0880] 1 H NMR (400MHz, DMSO-d6) δ12.64(s,1H),10.10(s,1H),8.21(t,J=5.2Hz,1H),7.79(d,J=9.0Hz, 1H),7.47(s,1H),7.44–7.36(m,1H),7.24(dd,J=8.6,1.6Hz,1H),7.08–6.89(m,3H),6.25(dd ,J=9.0,2.3Hz,1H),6.10(d,J=2.3Hz,1H),4.66(dd,J=7.7,6.0Hz,2H),4.32(t,J=5.9Hz,2H) ,4.02(s,2H),3.43(dd,J=7.2,5.2Hz,2H),3.33–3.10(m,5H),2.49–2.36(m,4H),2.22(s,3H).

[0881] Embodiment 48

[0882] Preparation of N-(5-(3-fluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((oxetan-3-ylmethyl)amino)benzamide 48

[0883]

[0884] Using the same preparation method as Example 47 and using 3-fluorobromobenzene as raw material, Compound 48 can be obtained.

[0885] MS m / z(ESI):529[M+H] +

[0886] 1 H NMR (400MHz, DMSO-d6) δ12.62(s,1H),10.09(s,1H),8.21(t,J=5.3Hz,1H),7.79(d,J=9.0Hz,1H),7.45(s, 1H),7.39(dd,J=8.6,0.8Hz,1H),7.34–7.25(m,1H),7.22(dd,J=8.6,1.6Hz,1H),7.11–7.02(m,2H),7.01–6 .95(m,1H),6.26(dd,J=9.0,2.3Hz,1H),6.10(d,J=2.4Hz,1H),4.67(dd,J=7.7,6.0Hz,2H),4.32(t,J=5.9H z,2H),4.02(s,2H),3.44(dd,J=7.2,5.2Hz,2H),3.31-3.25(m,4H),3.21(m,1H),2.45(m,4H),2.24(s,3H).

[0887] Embodiment 49

[0888] Preparation of N-(5-(2,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((oxetan-3-ylmethyl)amino)benzamide 49

[0889]

[0890] Using the same preparation method as Example 47 and using 3-fluorobromobenzene as raw material, Compound 49 can be obtained.

[0891] MS m / z(ESI):547[M+H] +

[0892] 1H NMR (400MHz, DMSO-d6) δ12.64(s,1H),10.10(s,1H),8.21(t,J=5.2Hz,1H),7.79(d,J=9.0Hz,1 H),7.53–7.35(m,2H),7.28–7.12(m,3H),7.12–7.01(m,1H),6.26(dd,J=9.0,2.3Hz,1H),6.10 (d,J=2.4Hz,1H),4.67(dd,J=7.7,5.9Hz,2H),4.32(t,J=5.9Hz,2H),4.02(s,2H),3.44(dd,J= 7.2,5.2Hz,2H),3.29(t,J=5.0Hz,4H),3.25–3.13(m,1H),2.44(t,J=5.0Hz,4H),2.23(s,3H).

[0893] Embodiment 50

[0894] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 50

[0895]

[0896] By adopting the same preparation method as Example 21 and using 2,5-difluorobromobenzene as raw material, compound 50 can be obtained.

[0897] MS m / z(ESI):593[M+H] + .

[0898] 1H NMR (400MHz, DMSO-d6) δ12.67(s,1H),10.07(s,1H),8.28(d,J=7.7Hz,1H),7.76(d,J=9.0Hz,1H),7.50(d,J=1.5Hz,1H ),7.43–7.34(m,2H),7.29(dd,J=8.7,1.6Hz,1H),7.19(td,J=9.3,4.7Hz,1H),7.15–7.04(m,1H),6.21(dd,J=9.0,2.3H z,1H),6.10(d,J=2.3Hz,1H),5.10-4.86m,2H),4.72(dt,J=14.8,7.3Hz,1H),3.78(dt,J=11.6,4.1Hz,2H),3.65(q,J= 8.7Hz,1H),3.46(tt,J=11.8,2.3Hz,2H),3.26(s,4H),2.46(s,4H),2.25(s,3H),1.96–1.81(m,2H),1.35-1.22(m,2H).

[0899] Embodiment 51

[0900] Preparation of N-(5-(1-(3-fluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 51

[0901]

[0902] By adopting the same preparation method as Example 21 and using 3-fluorobromobenzene as raw material, Compound 51 can be obtained.

[0903] MS m / z(ESI):575[M+1] + .

[0904] 1H NMR (400MHz, DMSO-d6) δ12.65(s,1H),10.06(s,1H),8.27(d,J=7.7Hz,1H),7.76(d,J=9.1Hz,1H),7.53(d,J=1.6H z,1H),7.39(d,J=8.7Hz,1H),7.35–7.26(m,2H),7.22–7.14(m,2H),7.01(td,J=8.7,2.3Hz,1H),6.21(dd,J=9.0,2 .3Hz,1H),6.10(d,J=2.3Hz,1H),5.10-4.75(m,2H),4.55(dt,J=14.8,7.1Hz,1H),3.85–3.56(m,3H),3.52–3.41( m,2H),3.23(d,J=5.6Hz,4H),2.43(d,J=5.8Hz,4H),2.22(s,3H),1.89(dd,J=10.8,6.8Hz,2H),1.50–1.24(m,2H).

[0905] Embodiment 52

[0906] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-ethylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 52

[0907]

[0908] Using the same preparation method as Example 50 and using N-ethylpiperazine as raw material, Compound 52 can be obtained.

[0909] MS m / z(ESI):607[M+1] +

[0910] 1H NMR (400MHz, DMSO-d6) δ12.67(s,1H),10.08(s,1H),8.27(d,J=7.6Hz,1H),7.76(d,J=9.0Hz,1H),7.50(s,1H),7.44–7.37(m,2H),7 .29(dd,J=8.7,1.7Hz,1H),7.19(td,J=9.3,4.7Hz,1H),7.10(ddd,J=8.9,7.5,3.5Hz,1H),6.20(dd,J=9.0,2.3Hz,1H),6.09(d,J=2 .3Hz,1H),5.10–4.86(m,2H),4.72(dt,J=14.8,7.3Hz,1H),3.87–3.72(m,2H),3.64(dd,J=10.6,6.5Hz,1H),3.46(ddt,J=13.0,11. 2,2.2Hz,2H),3.25(s,4H),2.48(s,4H),2.34(q,2H),1.96–1.83(m,2H),1.30(dtd,J=13.5,9.8,4.0Hz,2H),1.01(t,J=7.1Hz,3H).

[0911] Embodiment 53

[0912] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-morpholinopiperidin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 53

[0913]

[0914] Using the same preparation method as Example 50 and using 4-(morpholin-4-yl)piperidine as raw material, Compound 53 can be obtained.

[0915] MS m / z(ESI):663[M+H] + .

[0916] 1H NMR (400MHz, DMSO-d6) δ12.68–12.62(m,1H),10.04(s,1H),8.28(d,J=7.6Hz,1H),7.74(d,J=9.1Hz,1H),7.50(d,J=1.5Hz ,1H),7.45–7.34(m,2H),7.29(dd,J=8.7,1.6Hz,1H),7.19(td,J=9.3,4.7Hz,1H),7.10(ddd,J=12.1,7.8,3.5Hz,1H),6.20 (dd,J=9.1,2.3Hz,1H),6.09(d,J=2.3Hz,1H),5.10-4.80(m,2H),4.72(dt,J=14.8,7.2Hz,1H),3.81(dd,J=29.3,12.1Hz,4 H),3.69–3.39(m,7H),3.30-3.25(m,2H),2.75(t,J=12.3Hz,2H),2.48-2.28(m,3H),2.01-1.78(m,4H),1.47–1.11(m,4H).

[0917] Embodiment 54

[0918] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 54

[0919]

[0920] Using the same preparation method as Example 50 and using 1-(tetrahydro-2H-pyran-4-yl)piperazine as raw material, Compound 54 can be obtained.

[0921] MS m / z(ESI):663[M+H] + .

[0922] 1H NMR (400MHz, DMSO-d6) δ12.67(s,1H),10.08(s,1H),8.27(d,J=7.6Hz,1H),7.76(d,J=9.0Hz,1H),7.50(s,1H),7.45–7.35(m,2H),7.29( dd,J=8.7,1.6Hz,1H),7.19(td,J=9.3,4.7Hz,1H),7.11(tt,J=8.9,3.5Hz,1H),6.20(dd,J=8.9,2.2Hz,1H),6.09(d,J=2.4Hz,1H),5.10 -4.80(m,2H),4.72(dt,J=14.9,7.3Hz,1H),3.87(dd,J=10.7,4.1Hz,2H),3.78(d,J=11.9Hz,2H),3.64(d,J=8.4Hz,1H),3. 52–3.39(m,2H),3.30–3.14(m,6H),2.58(s,4H),2.38(s,1H),1.96–1.84(m,2H),1.71(d,J=12.4Hz,2H),1.46–1.21(m,4H).

[0923] Embodiment 55

[0924] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-cyclopropylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 55

[0925]

[0926] Using the same preparation method as Example 50 and using 4-cyclopropylpiperazine as raw material, Compound 55 can be obtained.

[0927] MS m / z(ESI):619[M+1] +

[0928] 1H NMR (400MHz, DMSO-d6) δ12.70(s,1H),10.11(s,1H),8.31(d,J=7.6Hz,1H),7.79(d,J=9.1Hz,1H),7.53(s,1H),7.47–7. 39(m,2H),7.32(dd,J=8.7,1.6Hz,1H),7.29–7.10(m,2H),6.23(dd,J=9.1,2.2Hz,1H),6.12(d,J=2.3Hz,1H),5.13–4.86 (m,2H),4.75(dt,J=14.8,7.3Hz,1H),3.81(d,J=11.7Hz,2H),3.75–3.62(m,1H),3.49(t,J=11.0Hz,2H),3.22(t,J=5.0H z,4H),2.65(t,J=4.9Hz,4H),1.93(d,J=12.5Hz,2H),1.66(tt,J=6.7,3.7Hz,1H),1.40–1.27(m,2H),0.48–0.32(m,4H).

[0929] Embodiment 56

[0930] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-trifluoromethylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 56

[0931]

[0932] Using the same preparation method as Example 50 and using 4-trifluoromethylpiperazine as raw material, Compound 56 can be obtained.

[0933] MS m / z(ESI):647[M+1] +

[0934] 1H NMR (400MHz, DMSO-d6) δ12.68(s,1H),10.12(s,1H),8.28(d,J=7.7Hz,1H),7.79(d,J=9.0Hz,1H),7.50(d,J=1.6H z,1H),7.44–7.36(m,2H),7.29(dd,J=8.8,1.6Hz,1H),7.22–7.17(m,1H),7.10(td,J=8.2,3.7Hz,1H),6.26–6.21( m,1H),6.15(d,J=2.3Hz,1H),5.14–4.83(m,2H),4.72(dt,J=14.7,7.1Hz,1H),3.79(d,J=11.6Hz,2H),3.65(s,1H ),3.46(t,J=10.9Hz,2H),3.33(t,J=5.0Hz,4H),2.98(t,J=5.0Hz,4H),1.92(t,J=13.1Hz,2H),1.36–1.26(m,2H).

[0935] Embodiment 57

[0936] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 57

[0937]

[0938] Using the same preparation method as Example 50 and using N-methyl-2-(pyrrolidin-1-yl)ethylamine as raw material, Compound 57 can be obtained.

[0939] MS m / z(ESI):621(M+H) +

[0940] 1H NMR (400MHz, Methanol-d4) δ7.72–7.59(m,2H),7.42(dd,J=8.8,0.8Hz,1H),7.30(dd,J=8.9,1.6Hz,1H),7.18(ddd,J=9. 1,5.7,3.1Hz,1H),7.07(td,J=9.3,4.5Hz,1H),6.98(ddd,J=12.2,8.3,3.6Hz,1H),6.14(dd,J=9.2,2.4Hz,1H),5.96(d, J=2.3Hz,1H),5.09–4.88(m,2H),4.84–4.71(m,1H),3.93(dt,J=11.7,4.1Hz,2H),3.68(dt,J=9.4,5.1Hz,1H),3.57(td, J=9.7,9.2,3.0Hz,4H),3.02(s,3H),2.80–2.58(m,6H),2.04(d,J=13.3Hz,2H),1.85(t,J=4.5Hz,4H),1.59-1.48(m,2H).

[0941] Embodiment 58

[0942] Preparation of N-(5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-((1R,4R)-5-methyl-2,5-diazabicyclo[2.2.10.1]heptan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 58

[0943]

[0944] Using the same preparation method as Example 39 and using (1R, 4R)-2-methyl-2,5-diazabicyclo[2.2.1]heptane as the raw material, Compound 58 can be obtained.

[0945] MS m / z(ESI):574[M+1] +

[0946] 1H NMR (400MHz, DMSO-d6) δ13.14(s,1H),10.19(s,1H),8.90(d,J=1.2Hz,1H),8.31(d,J=7.5Hz,1H ),7.75(d,J=9.0Hz,1H),7.53(s,1H),7.07–6.95(m,3H),5.93(d,J=8.7Hz,1H),5.76(s,1H),4.4 2(s,1H),4.19(s,2H),3.82(d,J=11.4Hz,2H),3.64(s,1H),3.48(t,J=10.8Hz,4H),3.32-3.28( m,2H),2.83(d,J=9.3Hz,1H),2.30(s,3H),1.92(q,J=12.5,12.0Hz,4H),1.35(d,J=11.5Hz,2H).

[0947] Embodiment 59

[0948] Preparation of N-(5-(3,5-difluorobenzyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-4-((1s,4s)-5-methyl-2,5-diazabicyclo[2.2.10.1]heptan-2-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 59

[0949]

[0950] Using the same preparation method as Example 39 and using (1S,4S)-2-methyl-2,5-diazabicyclo[2.2.1]heptane as raw material, Compound 59 can be obtained.

[0951] MS m / z(ESI):574[M+1] +

[0952] 1H NMR (400MHz, DMSO-d6) δ13.14(s,1H),10.19(s,1H),8.90(d,J=1.2Hz,1H),8.31(d,J=7.5Hz,1H ),7.75(d,J=9.0Hz,1H),7.53(s,1H),7.07–6.95(m,3H),5.93(d,J=8.7Hz,1H),5.76(s,1H),4.4 2(s,1H),4.19(s,2H),3.82(d,J=11.4Hz,2H),3.64(s,1H),3.48(t,J=10.8Hz,4H),3.32-3.28( m,2H),2.83(d,J=9.3Hz,1H),2.30(s,3H),1.92(q,J=12.5,12.0Hz,4H),1.35(d,J=11.5Hz,2H).

[0953] Embodiment 60

[0954] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-(2-(dimethylamino)ethyl)-1-carbonyl-6-((tetrahydro-2H-pyran-4-yl)amino)isoindoline-5-carboxamide 60

[0955]

[0956]

[0957] Step 1: Dimethyl 2-((tetrahydro-2H-pyran-4-yl)amino)terephthalate 60b

[0958] To a dry reaction flask, 60a (2.09 g, 10 mmol) and 50 ml of DCM were added, followed by sodium acetate borohydride (4.24 g, 20 mmol), 4-tetrahydropyrone (2.2 g, 22 mmol) and 5 mL of acetic acid. The reaction was stirred at room temperature overnight, and TLC showed that the reaction was complete. Potassium carbonate solution was added and stirred for 30 minutes, the organic phase was separated, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a crude product 60b which was used directly in the next step.

[0959] MS m / z(ESI):294[M+H]+

[0960] Step 2 Dimethyl 2-bromo-5-((tetrahydro-2H-pyran-4-yl)amino)terephthalate 60c

[0961] Add 60b (2.93 g, 10.0 mmol) and 100 ml of acetonitrile to the reaction flask, then add NBS (1.78 g, 10.0 mmol), and stir overnight at room temperature. TLC shows that the starting material has reacted. Concentrate the reaction solution under reduced pressure, pour into water, stir for 0.5 hours, filter and dry the resulting solid, which is 60c (3.2 g, 8.6 mmol).

[0962] MS m / z(ESI):373[M+2]+

[0963] 1H NMR(400MHz,DMSO-d6)δ7.97(s,1H),7.77–7.69(d,J=8.0,1H),7.17(s,1H),3.85(s,3H), 3.84(s,3H),3.78-3.66(m,1H),3.53-3.41(m,2H),1.96-1.85(m,2H),1.50-1.33(m,2H).

[0964] Step 3 Dimethyl 2-cyano-5-((tetrahydro-2H-pyran-4-yl)amino) terephthalate 60d

[0965] Add 60c (1.86 g, 5 mmol), CuCN (890 mg, 10 mmol) and DMF (30 ml) to a single-mouth bottle, heat to 150 ° C, maintain at this temperature for 1 hour, cool to room temperature, pour into ethyl acetate-ammonia system and stir, clarify and separate. The organic phase is washed three times with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 60d (1.27 g, 4.0 mmol).

[0966] MS m / z(ESI):319[M+H]+

[0967] Step 4: Methyl 1-carbonyl-6-((tetrahydro-2H-pyran-4-yl)amino)isoindoline-5-carboxylate 60e

[0968] 60d (319 mg, 1.0 mmol) was dissolved in THF-MeOH system, 100 mg Raney-Ni was added, and the mixture was stirred overnight under hydrogen atmosphere. TLC showed that the reaction was complete, and the filtrate was filtered and concentrated under reduced pressure. The obtained solid was slurried with EtOAc-PE system to obtain 60e (230 mg, 0.8 mmol).

[0969] MS m / z(ESI):291[M+H]+

[0970] Step 5: Methyl 2-(2-bromoethyl)-1-carbonyl-6-((tetrahydro-2H-pyran-4-yl)amino)isoindoline-5-carboxylate 60f

[0971] Compound 60e (1.45 g, 5.0 mmol) was dissolved in DMF, NaH (0.8 g, 60%, 20 mmol, 4 eq) was added and stirred at room temperature for 0.5 h, then dibromoethane (9.25 g, 50 mmol) was added and stirred at room temperature for 4 h, poured into an aqueous ammonium chloride solution, extracted three times with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, silica gel was added and mixed, and flash purified (DCM: MeOH = 20: 1) to obtain 60f (0.79 g, 2.0 mmol).

[0972] MS m / z(ESI):433[M+H]+

[0973] Step 6: Methyl 2-(2-(dimethylamino)ethyl)-1-carbonyl-6-((tetrahydro-2H-pyran-4-yl)amino)isoindoline-5-carboxylate 60g

[0974] Add 60f (433 mg, 1.0 mmol), dimethylamine hydrochloride (400 mg, 5.0 mmol), potassium carbonate (1.38 g, 10.0 mmol) and DMF (20 ml) to a dry reaction bottle. After stirring overnight, add dichloromethane to dilute the reaction solution, filter the filtrate, concentrate under reduced pressure to remove the high-boiling DMF, and flash purify (DCM: MeOH = 20: 1) to obtain 60g (180 mg, 0.5 mmol).

[0975] MS m / z(ESI):362[M+H]+

[0976] Step 7: 2-(2-(dimethylamino)ethyl)-1-carbonyl-6-((tetrahydro-2H-pyran-4-yl)amino)isoindoline-5-carboxylic acid 60h

[0977] Add compound 60g (361mg, 1.0mmol), 10ml tetrahydrofuran, and then add 5ml lithium hydroxide solution (2M) to a dry reaction bottle, and react at 80°C for 2 hours. TLC shows that the raw materials have reacted. Concentrate the reaction solution under reduced pressure to remove most of the tetrahydrofuran, add hydrochloric acid to neutralize the reaction solution, and use preparative HPLC to obtain 60h (300mg, 0.86mmol).

[0978] MS m / z(ESI):348[M+H]+

[0979] Step 8 2-(2-(dimethylamino)ethyl)-1-carbonyl-6-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetylamino)isoindoline-5-carboxylic acid 60i

[0980] 60h (200 mg, 0.57 mmol), dichloromethane and 0.5 ml of trifluoroacetic anhydride were added to the reaction flask. After a few minutes, the reaction solution turned from turbid to clear, indicating the completion of the reaction. The reaction solution was concentrated under reduced pressure, and a small amount of water was added to the residue. The solution was concentrated to dryness using a high vacuum pump to obtain 60i (252 mg, 0.57 mmol) as a solid.

[0981] MS m / z(ESI):444[M+H]+

[0982] Step 9: N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-(2-(dimethylamino)ethyl)-1-carbonyl-6-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)isoindoline-5-carboxamide 60j

[0983] Add 60i (221 mg, 0.5 mmol), 25 ml of dichloromethane and 1 ml of oxalyl chloride to a dry reaction flask. Stir at room temperature for 4-6 hours, concentrate the reaction solution to dryness under reduced pressure, dilute with tetrahydrofuran, cool to -40°C, and protect with nitrogen. Then add 1 ml of DIPEA, and then slowly add a THF solution of I-2 (130 mg, 0.5 mmol). After the addition, naturally warm to room temperature. TLC shows that most of the raw materials have reacted. Concentrate the reaction solution to an appropriate volume, and then purify it by preparative thin layer chromatography (DCM: MeOH = 10: 1) to obtain 60j (171 mg, 0.25 mmol, 50%).

[0984] MS m / z(ESI):685[M+H]+

[0985] Step 10: N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-(2-(dimethylamino)ethyl)-1-carbonyl-6-((tetrahydro-2H-pyran-4-yl)amino)isoindoline-5-carboxamide 60

[0986] 13051l (120mg, 0.175mmol) and 10ml methanol were added to the reaction flask, and 0.5ml potassium carbonate solution was added after dissolution. After 10min, TLC showed that the raw material had reacted completely. The methanol was concentrated under reduced pressure, and ethyl acetate was added for extraction, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by preparative thin layer chromatography (DCM:MeOH=10:1) to obtain 60 (70mg, 0.12mmol).

[0987] MS m / z(ESI):589[M+H]+

[0988] 1H NMR(400MHz,Chloroform-d)δ10.86(s,1H),9.85(s,1H),7.77(s,1H),7.63(s,1H),7.57 (d,J=7.3Hz,1H),7.32(d,J=8.7Hz,1H),7.17(dd,J=8.6,1.6Hz,1H),7.01(s,1H),6.76–6 .67(m,2H),6.65–6.56(m,1H),4.29(s,2H),4.04(s,2H),3.95(m,2H),3.72(t,J=6.2Hz,2 H),3.49(m,3H),2.63(t,J=6.2Hz,2H),2.33(s,6H),1.98(d,J=13.3Hz,2H),1.52(m,2H).

[0989] Embodiment 61

[0990] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(N-methyl-2-morpholinoacetylamino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 61

[0991]

[0992] Step 1 N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(methylamino)-2-nitrobenzamide 61a

[0993] Compound 11b (500 mg, 1.17 mmol), methylamine hydrochloride (157 mg, 2.3 mmol) and DIPEA (603 mg, 4.68 mmol) were dissolved in DMSO (10 mL). The mixture was stirred at 130°C for 3 hours, cooled, quenched with water, extracted with ethyl acetate, the organic phases were combined, washed twice with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography (DCM: MeOH = 15: 1) to obtain 61a 400 mg as a yellow solid.

[0994] MS m / z(ESI):438[M+H] +

[0995] Step 2 N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(N-methyl-2-morpholinoacetylamino)-2-nitrobenzamide 61b

[0996] Compound 61a (150 mg, 0.34 mmol), 2-morpholinoacetic acid (50 mg, 0.34 mmol), HATU (130 mg, 0.34 mmol) and triethylamine (68 mg, 0.68 mmol) were dissolved in DMF (10 mL). The mixture was stirred at room temperature for 2 hours, quenched by adding water, extracted with ethyl acetate, the organic phases were combined, washed twice with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography (DCM: MeOH = 10: 1) to give 61b (120 mg) as a yellow solid.

[0997] MS m / z(ESI):565[M+H] + .

[0998] Step 3 2-amino-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(N-methyl-2-morpholinoacetamido)benzamide 61c

[0999] Disperse 61b (120 mg, 0.21 mmol) N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(N-methyl-2-morpholinoacetylamino)-2-nitrobenzamide in ethyl acetate (10 mL), add 10% Pd / C (15 mg), and stir the mixture under hydrogen at room temperature for 2 hours, filter, and concentrate. The crude product 61c is used directly in the next step.

[1000] MS m / z(ESI):535[M+H] + .

[1001] Step 4: N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(N-methyl-2-morpholinoacetylamino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 61

[1002] Tetramethylammonium triacetoxyborohydride (86 mg, 0.73 mmol) was slowly added to a DCM (3 mL) solution of compound 61c (60 mg, 0.11 mmol), tetrahydro-4H-pyran-4-one (16 mg, 0.16 mmol) and trifluoroacetic acid (1 mL), and the mixture was stirred at room temperature for 2 hours. Saturated aqueous NaHCO3 solution was added to quench the reaction, extracted with DCM, the organic phases were combined, washed twice with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography to obtain 61 (4 mg) as a white solid.

[1003] MS m / z(ESI):619[M+H] + .

[1004] 1 H NMR(400MHz,Chloroform-d)δ8.34(d,J=8.7Hz,1H),8.17(d,J=7.4Hz,1H),8.12(s ,1H),7.81(s,1H),7.44(d,J=8.8Hz,1H),7.35(d,J=8.7Hz,1H),6.65(dd,J=33.6,8 .1Hz,3H),5.92(d,J=8.8Hz,1H),5.79(s,1H),4.09–3.91(m,6H),3.81(s,4H),3.62 -3.50(m,3H),2.89(s,3H),2.74(s,4H),2.03(d,J=13.1Hz,2H),1.62-1.51(m,2H).

[1005] Embodiment 62

[1006] Preparation of N-(5-(1-(3,5-difluorophenyl)-2,2,2-trifluoro-ethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 62

[1007]

[1008] First step 1-(3,5-difluorophenyl)-2,2,2-trifluoroethyl-1-ol 62b

[1009] To a DMF (50 mL) solution of compound 62a (5.0 g, 35.2 mmol), CF3-TMS (6.0 g, 42.3 mmol) and potassium carbonate (9.7 g, 70.4 mmol) were added. The reaction solution was stirred overnight at room temperature. Water was added to the reaction solution, and then extracted with ethyl acetate (200 mL). The organic phase was washed three times with saturated brine, dried and spin-dried to obtain the crude product, which was purified by column chromatography to obtain compound 62b (7.0 g, 33.0 mmol).

[1010] Step 2 1-(3,5-difluorophenyl)-2,2,2-trifluoroethyl 4-methylbenzenesulfonate 62c

[1011] Compound 62b (7 g, 33.0 mmol) was dissolved in tetrahydrofuran (100 mL), NaH (1.6 g, 39.6 mmol) was added in an ice bath, stirred for 0.5 h, and Tos-Cl (8.1 g, 39.6 mmol) was slowly added. The reaction solution was stirred at room temperature for 2 hours, poured into ice water, extracted with ethyl acetate (200 mL), and the organic phase was washed with saturated sodium bicarbonate solution and brine, dried with anhydrous sodium sulfate, filtered and spin-dried to obtain the initial product, which was purified by silica gel column chromatography to obtain compound 62c (7.0 g, 19.1 mmol).

[1012] Step 3 5-(1-(3,5-difluorophenyl)-2,2,2-trifluoroethyl)-2-fluorobenzonitrile 62e

[1013] Compound 62c (1 g, 2.73 mmol) was dissolved in 1,4-dioxane (15 mL), and compound 62d (900 mg, 5.46 mmol), Xphos-Pd-G2 (110 mg, 0.14 mmol) and potassium phosphate (926 mg, 4.37 mmol) were added, and the reaction solution was reacted at 100°C under microwave for 6 hours, with N2 protection. After cooling, the reaction solution was poured into water (50 mL), extracted with ethyl acetate (50 mL), and the organic phase was washed with saturated brine, dried and spin-dried to obtain a crude product, which was purified by column chromatography to obtain compound 62e (200 mg, 0.63 mmol).

[1014] Step 4 5-(1-(3,5-difluorophenyl)-2,2,2-trifluoro-ethyl)-1H-indazol-3-amine 62f

[1015] Compound 62e (400 mg, 1.26 mmol) and hydrazine hydrate (313 mg, 6.3 mmol) were mixed in N-methylpyrrolidone (6 mL), and the reaction solution was reacted at 120 ° C in a microwave reactor for 1 hour. After cooling, it was added to (30 mL) and extracted with ethyl acetate (50 mL). The organic phase was dried and spin-dried to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 62f (250 mg, 0.76 mmol).

[1016] MS m / z(ESI):328[M+H] + .

[1017] Step 5: N-(5-(1-(3,5-difluorophenyl)-2,2,2-trifluoro-ethyl)-1H-indazol-3-yl)-4-(4-methylpiperazine-1-yl)-2-(2-, 2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamido)benzamide 62g

[1018] Compound 62f (500 mg, 1.2 mmol) was suspended in dichloromethane (30 mL), cooled to 0°C, oxalyl chloride (254 mg, 2.0 mmol) was added dropwise to the solution, the reaction solution was stirred at room temperature overnight, spin-dried, and then dichloromethane (10 mL) was added and then spin-dried. The product was dissolved in tetrahydrofuran (5 mL), cooled to -20°C, and a solution of compound 1f (250 mg, 0.76 mmol) and DIPEA (196 mg, 1.52 mmol) in tetrahydrofuran (15 mL) was added dropwise to the above coolant. The reaction was stirred at -20°C for 2 hours, and then continued to stir at room temperature for 2 hours. The reaction solution was diluted with dichloromethane (50 mL), washed with mixed brine, dried and spin-dried to obtain a crude product, which was purified by column chromatography (DCM / MeOH=10:1) to obtain compound 62g (50 mg, 0.069 mmol).

[1019] MS m / z(ESI):725[M+H] + .

[1020] Step 6: N-(5-(1-(3,5-difluorophenyl)-2,2,2-trifluoro-ethyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 62

[1021] Compound 62g (50 mg, 0.069 mmol) was dissolved in a mixed solvent of methanol (10 mL) and water (5 mL), potassium carbonate (90.7 mg, 0.66 mmol) was added, the reaction solution was stirred at room temperature for 3 hours, water (20 ml) was added, and it was extracted with dichloromethane (50 ml). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 62 (10 mg, 0.016 mmol).

[1022] MS m / z(ESI):629[M+H]+.

[1023] 1H NMR (400MHz, DMSO-d6) δ12.78(s,1H),10.17(s,1H),8.28(d,J=7.9Hz,1H),7.76–7.74(m ,1H),7.48–7.45(m,1H),7.41–7.37(m,1H),7.27–7.11(m,4H),6.23–6.19(m,1H),6.12–6 .08(m,1H),5.50–5.44(m,1H),3.81–3.73(m,2H),3.69–3.63(m,1H),3.50–3.42(m,2H), 3.25–3.20(m,4H),2.42–2.36(m,4H),2.19(s,3H),1.93–1.87(m,2H),1.32–1.26(m,2H).

[1024] Embodiment 63

[1025] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 63

[1026]

[1027] By adopting the same preparation method as Example 43 and using 3,5-difluorobromobenzene as raw material, Compound 63 can be obtained.

[1028] MS m / z(ESI):603[M+H]+.

[1029] 1H NMR (400MHz, Methanol-d4) δ7.70(d,J=9.0Hz,1H),7.57(d,J=1.4Hz,1H),7.39(dd,J=8.7,0.8Hz,1H),7.25(dd, J=8.5,1.6Hz,1H),7.04(td,J=9.1,4.6Hz,1H),6.98–6.86(m,2H),6.12(dd,J=9.1,2.4Hz,1H),5.95(d,J=2.4Hz, 1H),4.06(s,2H),3.93(dt,J=11.9,4.1Hz,2H),3.67(dd,J=9.0,4.7Hz,1H),3.62–3.51(m,4H),3.45(q,J=7.0Hz ,2H),2.86–2.69(m,6H),2.03(d,J=13.5Hz,2H),1.87(t,J=5.4Hz,4H),1.60–1.45(m,3H),1.19(t,J=7.0Hz,3H).

[1030] Embodiment 64

[1031] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(3-fluoropyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 64

[1032]

[1033] Step 1 tert-Butyl 4-(ethyl(2-hydroxyethyl)amino)-2-nitrobenzoate 64b

[1034] Compound I-1b (3.0 g, 12.4 mmol, 1.0 eq) and 2-(ethylamino)ethane-1-ol (64a) (3.3 g, 37.2 mmol, 3.0 eq) were placed in a mixed solvent of NMP (10 ml), and DIPEA (8.6 g, 49.6 mmol, 4.0 eq) was added, and the mixture was reacted at 105° C. for 8 h.

[1035] The mixture was cooled to room temperature, ethyl acetate (80 ml) and H2O (20 ml) were added for extraction, and the organic layer was washed with brine (20 ml), dried over anhydrous sodium sulfate, concentrated, and purified by flash column to obtain a yellow oil 64b (4.4 g).

[1036] MS m / z(ESI):311[M+1] +

[1037] Step 2 tert-Butyl 4-(ethyl(2-oxoethyl)amino)-2-nitrobenzoate 64c

[1038] Compound 64b (3.4 g, 11.0 mmol, 1.0 eq) was dissolved in DCM (80 ml), dess-martin (5.6 g, 13.2 mmol, 1.2 eq) was added, and the mixture was reacted at room temperature for 3 h. Sodium bicarbonate and sodium thiosulfate aqueous solution were added to quench the mixture, and the organic layer was separated, dried over anhydrous sodium sulfate, and concentrated to obtain 64c which was directly used in the next step.

[1039] MS m / z(ESI):309[M+1] +

[1040] Step 3 4-(2-(3-(3-fluoropyrrolidin-1-yl)ethyl)amino)-2-nitrobenzoic acid tert-butyl ester 64d

[1041] 3-Fluoropyrrolidine hydrochloride (1.1 g, 8.7 mmol, 1.5 eq) was added to DCM (50 ml), and DIPEA (1.5 ml, 8.7 mmol, 1.5 eq) and compound 64c (1.8 g, 5.8 mmol, 1.0 eq) were added in sequence. After reacting at room temperature for 30 min, sodium acetate borohydride (3.7 g, 17.4 mmol, 3.0 eq) was added and the reaction was continued for 1 h.

[1042] After neutralization by adding aqueous sodium bicarbonate solution, the mixture was extracted with DCM (50 ml×2). The organic layers were combined, dried over anhydrous sodium sulfate, concentrated, and purified by flash column to obtain a yellow oil 64d (1.7 g).

[1043] MS m / z(ESI):382[M+1] +

[1044] Step 4: N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(3-fluoropyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 64

[1045] Using the same preparation method as Example 43, compound 63 can be obtained by replacing 43a with 64d as the starting material.

[1046] MS m / z(ESI):621[M+1] +

[1047] 1H NMR (400MHz, DMSO-d6) δ12.62(s,1H),9.97(s,1H),8.35(d,J=7.3Hz,1H),7.77(d,J=9.1Hz,1H),7.47(s,1H),7.43–7.36(m, 1H),7.25(dd,J=8.6,1.6Hz,1H),7.06–6.92(m,3H),5.99(dd,J=9.1,2.3Hz,1H),5.83(d,J=2.3Hz,1H),5.21(dt,J=56.1,5. 6Hz,1H),4.03(s,2H),3.83(dt,J=11.7,3.9Hz,2H),3.65–3.53(m,1H),3.51–3.36(m,5H),2.99–2.82(m,2H),2.74–2.55(m, 3H),2.36(q,J=7.9Hz,1H),2.14(ddq,J=27.6,14.1,6.8Hz,1H),2.01–1.78(m,3H),1.45–1.28(m,2H),1.12(t,J=6.9Hz,3H).

[1048] Embodiment 65

[1049] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(pyrrolidin-1-yl)ethyl)thio)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 65

[1050]

[1051] Using the same preparation method as Example 44 and using 2-(1-pyrrolidinyl)ethanethiol as the raw material, Compound 65 can be obtained.

[1052] MS m / z(ESI):592(M+H) +

[1053] 1H NMR (400MHz, Methanol-d4) δ7.74(d,J=8.3Hz,1H),7.57(dd,J=1.6,0.8Hz,1H),7.41(dd,J=8.6,0.8Hz,1H),7. 24(dd,J=8.6,1.6Hz,1H),6.84–6.78(m,2H),6.75–6.67(m,2H),6.61(dd,J=8.3,1.8Hz,1H),4.06(s,2H),3.92 (dt,J=11.8,4.0Hz,2H),3.69(td,J=9.4,4.6Hz,1H),3.56(ddd,J=11.9,10.2,2.5Hz,2H),3.20–3.13(m,2H),2 .86–2.79(m,2H),2.72–2.64(m,4H),2.05–1.96(m,2H),1.88–1.80(m,4H),1.50(dtd,J=13.7,10.0,4.2Hz,2H).

[1054] Embodiment 66

[1055] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(3,3-difluoropyrrolidin-1-yl)ethyl)(ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 66

[1056]

[1057] Using the same preparation method as Example 64 and using 3,3-difluoropyrrolidine hydrochloride as raw material, Compound 66 can be obtained.

[1058] MS m / z(ESI):639[M+1] +

[1059] 1H NMR (400MHz, DMSO-d6) δ12.59(s,1H),9.95(s,1H),8.33(d,J=7.3Hz,1H),7.74(d,J=9.1Hz,1H),7.44(d,J=1.5Hz,1H),7.37(dd,J =8.6,0.8Hz,1H),7.22(dd,J=8.6,1.6Hz,1H),7.03–6.89(m,3H),5.96(dd,J=9.0,2.3Hz,1H),5.79(d,J=2.4Hz,1H),4.00(s,2H),3 .80(dt,J=11.6,4.0Hz,2H),3.63–3.51(m,1H),3.41(ddt,J=15.6,10.2,5.1Hz,6H),2.94(t,J=13.4Hz,2H),2.74(t,J=6.9Hz,2H) ,2.59(t,J=7.3Hz,2H),2.21(tq,J=13.8,6.8Hz,2H),1.97–1.87(m,2H),1.34(tdd,J=13.5,9.7,3.9Hz,2H),1.08(t,J=6.9Hz,3H).

[1060] Embodiment 67

[1061] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(3,4-difluoropyrrolidin-1-yl)ethyl)(ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 67

[1062]

[1063] Using the same preparation method as Example 64 and using 3,4-difluoropyrrolidine hydrochloride as raw material, Compound 67 can be obtained.

[1064] MS m / z(ESI):639[M+1] +

[1065] 1H NMR (400MHz, DMSO-d6) δ12.61(s,1H),9.97(s,1H),8.36(d,J=7.3Hz,1H),7.77(d,J=9.1Hz,1H),7.47(s,1H), 7.40(dd,J=8.5,0.8Hz,1H),7.25(dd,J=8.6,1.6Hz,1H),7.05–6.92(m,3H),5.99(dd,J=9.1,2.3Hz,1H),5.83 (d,J=2.4Hz,1H),5.27–5.03(m,2H),4.04(s,2H),3.83(dt,J=11.6,4.0Hz,2H),3.67–3.54(m,1H),3.52–3.35 (m,6H),3.07–2.82(m,4H),2.66(t,J=7.2Hz,2H),2.01–1.90(m,2H),1.45–1.31(m,2H),1.11(t,J=6.9Hz,3H).

[1066] Embodiment 68

[1067] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(3,4-dimethylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 68

[1068]

[1069] By adopting the same preparation method as Example 50 and using 1,2-dimethylpiperazine as raw material, Compound 68 can be obtained.

[1070] MS m / z(ESI):607[M+H] + .

[1071] 1H NMR (400MHz, DMSO-d6) δ12.67(s,1H),10.08(s,1H),8.30(d,J=7.7Hz,1H),7.76(d,J=9.1Hz,1H),7.50(d,J=1.6Hz,1H),7.45–7.35(m ,2H),7.30(dd,J=8.7,1.7Hz,1H),7.20(td,J=9.3,4.7Hz,1H),7.15–7.06(m,1H),6.22(dd,J=9.1,2.3Hz,1H),6.09(d,J=2.3Hz,1H),5 .10(t,J=8.7Hz,0H),5.03–4.93(m,1H),4.87(dd,J=9.3,6.4Hz,0H),4.73(dt,J=14.8,7.2Hz,1H),3.84–3.73(m,2H),3.69(d,J=13.8H z,3H),3.47(ddt,J=11.8,9.8,2.3Hz,2H),2.82(s,2H),2.22(s,4H),1.90(d,J=13.4Hz,2H),1.40–1.25(m,2H),1.05(d,J=6.1Hz,3H).

[1072] Embodiment 69

[1073] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(3-methyl-4-ethylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 69

[1074]

[1075] Using the same preparation method as Example 50 and using 1-ethyl-2-methylpiperazine as raw material, Compound 69 can be obtained.

[1076] MS m / z(ESI):621[M+H] + .

[1077] 1H NMR (400MHz, DMSO-d6) δ12.67(s,1H),10.07(s,1H),8.29(d,J=7.7Hz,1H),7.76(d,J=9.1Hz,1H),7.50(d,J=1.5Hz,1H),7.45–7.34(m,2H),7.30(d d,J=8.7,1.6Hz,1H),7.20(td,J=9.3,4.7Hz,1H),7.15–7.07(m,1H),6.2 1(dd,J=9.1,2.3Hz,1H),6.08(d,J=2.4Hz,1H),5.10(t,J=8.7Hz,1H),4.9 8(dd,J=9.1,6.2Hz,1H),4.87(dd,J=9.3,6.4Hz,1H),4.73(dt,J=14.8,7 .2Hz,1H),3.85–3.73(m,2H),3.72–3.53(m,3H),3.47(ddt,J=11.8,9.7,2 .3Hz,2H),2.95–2.68(m,3H),2.57(t,J=10.5Hz,1H),2.43(s,1H),2.29(d ,J=12.2Hz,2H),2.00–1.79(m,2H),1.39–1.25(m,2H),1.11–0.87(m,6H).

[1078] Embodiment 70

[1079] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(2,4-dimethylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 70

[1080]

[1081] By adopting the same preparation method as Example 50 and using 1,3-dimethylpiperazine as raw material, Compound 69 can be obtained.

[1082] MS m / z(ESI):607[M+H] + .

[1083] 1H NMR (400MHz, DMSO-d6) δ12.68(s,1H),10.07(s,1H),8.32(d,J=7.6Hz,1H),7.79 (d,J=9.1Hz,1H),7.54(d,J=1.5Hz,1H),7.46–7.37(m,2H),7.32(dd,J=8.7,1.7 Hz,1H),7.23(td,J=9.3,4.7Hz,1H),7.18–7.09(m,1H),6.20(dd,J=9.2,2.3Hz, 1H),6.06(d,J=2.3Hz,1H),5.13(t,J=8.7Hz,0H),5.01(dd,J=9.4,6.9Hz,1H),4. 90(dd,J=9.3,6.4Hz,1H),4.76(dt,J=14.7,7.2Hz,1H),4.15(s,1H),3.81(dd,J =11.6,3.5Hz,2H),3.66(dd,J=11.5,6.0Hz,1H),3.50(t,J=11.2Hz,3H),3.00(t d,J=12.0,3.5Hz,1H),2.84(d,J=10.9Hz,1H),2.69(d,J=11.1Hz,1H),2.20(s,4 H),2.07–1.84(m,1H),1.34(ddt,J=12.1,8.8,4.7Hz,2H),1.10(d,J=6.5Hz,3H).

[1084] Embodiment 71

[1085] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(3,3,4,4-tetrafluoropyrrolidin-1-yl)ethyl)(ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 71

[1086]

[1087] Using the same preparation method as Example 64 and using 3,3-difluoropyrrolidine hydrochloride as raw material, Compound 66 can be obtained.

[1088] MS m / z(ESI):639[M+1] +

[1089] 1H NMR (400MHz, DMSO-d6) δ12.61(s,1H),9.98(s,1H),8.36(d,J=7.3Hz,1H),7.77(d,J=9.1Hz,1H),7.47( s,1H),7.40(dd,J=8.6,0.8Hz,1H),7.25(dd,J=8.6,1.6Hz,1H),7.00(ddd,J=12.1,8.0,2.0Hz,3H),6.0 0(dd,J=9.0,2.3Hz,1H),5.82(d,J=2.4Hz,1H),4.04(s,2H),3.83(dt,J=11.6,4.0Hz,2H),3.51–3.37( m,6H),3.26(s,3H),2.70(t,J=7.2Hz,2H),1.99–1.89(m,2H),1.43–1.31(m,2H),1.11(t,J=6.9Hz,3H).

[1090] Embodiment 72

[1091] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)-4-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)benzamide 72

[1092]

[1093] Using the same preparation method as Example 50 and using 1,3-dimethylpiperazine as raw material, Compound 72 can be obtained.

[1094] MS m / z(ESI):661[M+H] + .

[1095] 1H NMR (400MHz, DMSO-d6) δ12.67(s,1H),10.08(s,1H),8.27(d,J=7.7Hz,1H),7.77(d,J=9.0Hz,1H),7.51(d,J=1.5Hz,1H),7.45–7.34(m,2H) ,7.29(dd,J=8.7,1.7Hz,1H),7.20(td,J=9.3,4.7Hz,1H),7.15–7.07(m,1H),6.21(dd,J=9.0,2.3Hz,1H),6.11(d,J=2.3Hz,1H),5.10(t,J= 8.7Hz,1H),4.98(dd,J=9.4,6.9Hz,1H),4.87(dd,J=9.3,6.4Hz,1H),4.73(dt,J=14.7,7.3Hz,1H),3.79(d,J=11.6Hz,2H),3.66(dd,J=10.9 ,6.6Hz,1H),3.47(ddt,J=13.3,11.5,2.4Hz,2H),3.28–3.13(m,7H),2.72(t,J=5.0Hz,4H),1.99–1.81(m,2H),1.32(qd,J=9.7,4.9Hz,2H).

[1096] Embodiment 73

[1097] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(2-methyl-4-ethylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 73

[1098]

[1099] Using the same preparation method as Example 50 and using 1-ethyl-3-methylpiperazine as raw material, Compound 73 can be obtained.

[1100] MS m / z(ESI):621[M+H] + .

[1101] 1H NMR(400MHz,DMSO-d6)δ12.72–12.63(m,1H),10.07(d,J=10.0Hz,1H),8.3 7–8.25(m,1H),7.84–7.70(m,1H),7.59–7.49(m,1H),7.48–7.36(m,2H),7 .36–7.28(m,1H),7.26–7.07(m,2H),6.23–6.15(m,1H),6.09–6.00(m,1H) ,5.16–5.07(m,1H),4.99(dd,J=9.8,6.9Hz,1H),4.87(dd,J=9.4,6.4Hz,0 H),4.73(dt,J=14.6,7.4Hz,1H),4.15(s,1H),3.79(d,J=11.7Hz,2H),3.6 5(d,J=7.5Hz,1H),3.55–3.40(m,3H),2.95(q,J=13.2,12.3Hz,2H),2.76( d,J=11.1Hz,1H),2.33(s,2H),2.15(d,J=10.9Hz,1H),1.95(d,J=20.6Hz, 3H), 1.34 (td, J=8.8, 8.3, 4.7Hz, 2H), 1.05 (tdd, J=14.0, 9.9, 6.9Hz, 6H).

[1102] Embodiment 74

[1103] 4-(Cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 74

[1104]

[1105]

[1106] Step 1 Synthesis of 4-fluoro-2-nitrobenzoic acid benzyl ester 74a

[1107] 4-Fluoro-2-nitrobenzoic acid (20 g, 108 mmol) was dissolved in DMF (150 mL), benzyl bromide (20.3 g, 119 mmol) was added dropwise, potassium carbonate (41.4 g, 300 mmol) was added and reacted at 90°C for 3-5 hours. Water was added to quench, and the mixture was extracted with dichloromethane, washed with water (100 mL) and saturated brine (100 mL) in sequence, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to obtain the title compound 74a (23 g, 83.6 mmol) with a yield of 77.4%.

[1108] Step 2 Synthesis of benzyl 2-nitro-4-((2-(pyrrolidin-1-yl)ethyl)amino)benzoate 74b

[1109] Compound 74a (14.3 g, 125.4 mmol) was dissolved in NMP (100 mL), 50 mL of diisopropylethylamine (DIPEA) was added and stirred for 0.5 hours, and then compound 13079-a (23 g, 83.6 mmol) was added and stirred at room temperature for 3-5 hours. Water was added to quench, and dichloromethane was extracted. The mixture was washed with water (150 mL) and saturated brine (150 mL) in sequence, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (DCM: MeOH = 98: 2) to obtain the title compound 74b (22 g, 59.6 mmol) with a yield of 71.3%.

[1110] MS m / z(ESI):370.2[M+H] + .

[1111] Step 3 Synthesis of benzyl 4-(cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-nitrobenzoate 74c

[1112] Compound 74b (20 g, 54.2 mmol) was dissolved in toluene (100 mL), and cyclopropylboronic acid (13.9 g, 162.6 mmol) and pyridine (12.8 g, 162.6 mmol) were added. Cu(OAc)2 (10.84 g, 54.2 mmol) and Cs2CO3 (8.8 g, 27.1 mmol) were added and stirred for 0.5 hours, and then heated to 140°C with an oxygen balloon for 8 hours. Water was added to quench, and dichloromethane was extracted. The mixture was washed with water (150 mL) and saturated brine (150 mL) in turn, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (DCM: MeOH = 96: 4) to obtain the title compound 74c (6.0 g, 14.7 mmol) with a yield of 27.1%.

[1113] MS m / z(ESI):410.3[M+H] + .

[1114] Step 4 Synthesis of benzyl 2-amino-4-(cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)benzoate 74d

[1115] Compound 74c (4.6 g, 11.2 mmol) was dissolved in a 5:1 mixture of methanol and water (25 mL), stirred evenly, and then zinc powder (13.8 g, 211 mmol) and ammonium chloride (13.8 g, 258 mmol) were added and stirred at room temperature for about 3 hours. The mixture was filtered and the filtrate was dried by spin drying. The crude product was purified by column chromatography (DCM: MeOH = 95: 5) to obtain the title compound 74d (3.0 g, 7.9 mmol) with a yield of 70.6%.

[1116] MS m / z(ESI):380.3[M+H] + .

[1117] Step 5 Synthesis of benzyl 4-(cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzoate 74e

[1118] Compound 74d (2.9 g, 7.6 mmol) was dissolved in DCM (25 mL), and 1 mL of trifluoroacetic acid was added under stirring. After stirring, tetrahydro-2H-pyran-4-one was added and stirred at room temperature for 0.5 hours. NaBH(OAc)3 was added and stirred for 5 hours. Saturated ammonium chloride solution was added dropwise to quench. The mixture was extracted with dichloromethane, washed with water (100 mL) and saturated brine (100 mL) for many times, dried over anhydrous sodium sulfate, filtered, concentrated, and column chromatography (DCM:MeOH=97:3) was performed to obtain the title compound 74e (2.0 g, 4.3 mmol) with a yield of 56.8%.

[1119] MS m / z(ESI):464.4[M+H] + .

[1120] Step 6 Synthesis of benzyl 4-(cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamide)benzoate 74f

[1121] Compound 74e (2.0 g, 4.3 mmol) was dissolved in anhydrous DCM (25 mL), and 4 mL of diisopropylethylamine (DIPEA) was added and stirred evenly. Trifluoroacetic anhydride was added dropwise under ice-water cooling, and then the temperature was slowly raised to room temperature and stirred for about 3 hours. Saturated ammonium chloride solution was added dropwise to quench. The mixture was extracted with dichloromethane, washed with water (100 mL) and saturated brine (100 mL) several times, dried over anhydrous sodium sulfate, filtered, concentrated, and column chromatography (DCM: MeOH = 98: 2) was performed to obtain the title compound 74f (1.0 g, 1.8 mmol) with a yield of 41.6%.

[1122] MS m / z(ESI):560.4[M+H] + .

[1123] Step 7 Synthesis of 74 g of 4-(cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamide)benzoic acid

[1124] Compound 74f (1.0 g, 1.8 mmol) was dissolved in methanol (20 mL), and Pd / C (1.0 g, 9.4 mmol) was added. After hydrogen was replaced three times, the mixture was reacted at room temperature for 1-3 hours under a hydrogen environment. After the reaction was complete, the mixture was filtered and the reaction solution was spin-dried for use to obtain a crude product 74 g (0.6 g, 1.27 mmol) with a yield of 71%.

[1125] MS m / z(ESI):470.4[M+H] + .

[1126] Step 8 Synthesis of 4-(cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamide)benzoyl chloride 74h

[1127] Compound 74g (0.6g, 1.27mmol) was dissolved in dichloromethane, and one drop of N,N-dimethylformamide (DMF) was added under stirring, followed by dropwise addition of oxalyl chloride (0.4g, 1.84mmol) and reaction at room temperature for 1-2 hours. After the reaction was complete, the mixture was dried and set aside to obtain the title compound 74h.

[1128] Step 9 Synthesis of 4-(cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-(2,2,2-trifluoro-N-(tetrahydro-2H-pyran-4-yl)acetamide)benzamide 74i

[1129] Compound I-5 (390 mg, 1.5 mmol) was dissolved in anhydrous DCM (15 mL), and 1 mL of diisopropylethylamine (DIPEA) was added and stirred evenly. Compound 74h (600 mg, 1.2 mmol) was added dropwise under ice-water cooling, and then the temperature was slowly raised to room temperature and stirred for about 1 hour. Saturated ammonium chloride solution was added dropwise to quench. Extraction with dichloromethane, drying over anhydrous sodium sulfate, filtration, concentration, and column chromatography (DCM: MeOH = 98: 2) gave the title compound 74i (240 mg, 0.33 mmol), with a yield of 22.5%.

[1130] MS m / z(ESI):711.4[M+H] + .

[1131] Step 10 Synthesis of 4-(cyclopropyl(2-(pyrrolidin-1-yl)ethyl)amino)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 74

[1132] Compound 74i (240 mg, 0.33 mmol) was dissolved in anhydrous methanol (10 mL), 1 mL of triethylamine was added and stirred evenly, and then refluxed at 80 ° C for 1-3 hours. Then the temperature was slowly raised to room temperature and stirred for about 1 hour. Saturated ammonium chloride solution was added dropwise to quench. Extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, concentrated, and column chromatography (DCM: MeOH = 95: 5) to obtain the title compound 74 (61 mg, 0.10 mmol), with a yield of 30%.

[1133] MS m / z(ESI):615.5[M+H] + .

[1134] Embodiment 75

[1135] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(1,4-dioxo-7-azaspiro[4.4]nonan-1-yl)ethyl)(ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 75

[1136]

[1137] Using the same preparation method as Example 64 and using 3-pyrrolidone ethylene acetal as raw material, Compound 75 can be obtained.

[1138] MS m / z(ESI):661[M+1] +

[1139] 1H NMR (400MHz, DMSO-d6) δ12.60(s,1H),9.96(s,1H),8.35(d,J=7.3Hz,1H),7.76(d,J=9.1Hz,1H) ,7.47(s,1H),7.40(d,J=8.6Hz,1H),7.24(dd,J=8.6,1.6Hz,1H),7.05–6.93(m,3H),5.99(dd,J= 9.0,2.3Hz,1H),5.82(d,J=2.4Hz,1H),4.04(s,2H),3.83(tt,J=6.3,2.7Hz,6H),3.59(s,1H),3. 51–3.35(m,6H),2.71–2.53(m,6H),2.02–1.85(m,4H),1.43–1.32(m,2H),1.11(t,J=6.9Hz,3H).

[1140] Embodiment 76

[1141] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(3-oxopyrrolidin-1-yl)ethyl)(ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 76

[1142]

[1143] Compound 75 (120 mg, 0.2 mmol) was added to 2-butanone (10 ml), 6M HCl (2.5 ml) was added, and the mixture was refluxed for 3 h, and the solvent was evaporated under reduced pressure. Sodium bicarbonate was added for neutralization, and the mixture was extracted with DCM (50 ml × 2). The organic layers were combined and subjected to column chromatography (DCM: MeOH = 98:2) to obtain compound 76 (40 mg).

[1144] MS m / z(ESI):617[M+1] +

[1145] 1H NMR (400MHz, DMSO-d6) δ12.58(s,1H),9.94(s,1H),8.33(d,J=7.3Hz,1H),7.74(d,J=9.1Hz,1H),7.44(s,1H),7. 37(d,J=8.6Hz,1H),7.22(dd,J=8.5,1.6Hz,1H),7.03–6.89(m,3H),5.98(dd,J=9.3,2.3Hz,1H),5.82(d,J=2.7Hz ,1H),4.01(s,2H),3.84–3.75(m,2H),3.57(d,J=8.7Hz,1H),3.50–3.35(m,5H),2.99(s,2H),2.91(t,J=6.9Hz,2H ),2.68(t,J=7.3Hz,2H),2.31(t,J=6.9Hz,2H),1.92(d,J=12.9Hz,2H),1.42–1.28(m,2H),1.10(t,J=6.9Hz,3H).

[1146] Embodiment 77

[1147] Preparation of N-(5-(2,5-difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(pyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 77

[1148]

[1149] Using the same preparation method as Example 46 and using 2,5-difluorobromobenzene as raw material, Compound 77 can be obtained.

[1150] MS m / z(ESI):589[M+H] + .

[1151] 1H NMR (400MHz, Methanol-d4) δ7.71(d,J=9.0Hz,1H),7.57(s,1H),7.40(dd,J=8.6,0.8Hz,1H),7.25(dd,J =8.8,1.6Hz,1H),7.05(td,J=9.0,4.5Hz,1H),6.99–6.87(m,2H),6.14(dd,J=9.1,2.4Hz,1H),5.97(d,J =2.3Hz,1H),4.06(s,2H),3.93(dt,J=11.7,4.0Hz,2H),3.69(dt,J=9.4,5.1Hz,1H),3.63–3.52(m,4H), 3.03(s,3H),2.80–2.63(m,6H),2.04(d,J=12.7Hz,2H),1.86(s,4H),1.53(ddd,J=18.7,9.3,4.5Hz,3H).

[1152] Embodiment 78

[1153] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(ethyl(2-(azetidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 78

[1154]

[1155] Using the same preparation method as Example 64 and using azetidine hydrochloride as raw material, Compound 78 can be obtained.

[1156] MS m / z(ESI):589[M+1] +

[1157] 1H NMR (400MHz, DMSO-d6) δ12.59(d,J=7.7Hz,1H),9.98(s,1H),8.35(d,J=7.4Hz,1H),7.75(d,J=9.0Hz ,1H),7.47–7.33(m,2H),7.22(dd,J=8.6,1.6Hz,1H),6.97(tdd,J=12.8,7.7,4.2Hz,3H),5.99(d,J= 9.3Hz,1H),5.82(d,J=2.4Hz,1H),4.01(s,2H),3.85–3.74(m,2H),3.60(s,3H),3.50–3.32(m,8H),2 .80–2.74(m,1H),2.14(s,2H),1.92(d,J=13.1Hz,2H),1.42–1.29(m,2H),1.08(q,J=7.6,7.0Hz,3H).

[1158] Embodiment 79

[1159] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-((2-(pyrrolidin-1-yl)ethyl)sulfinyl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 79

[1160]

[1161] Compound 65 (100 mg) was dissolved in DCM (10 ml), and m-CPBA (29 mg, 0.17 mmol, 1.0 eq) was added under ice-water bath cooling, and the reaction was continued for 4 h. Sodium bicarbonate aqueous solution was added for neutralization, and DCM (15 ml × 2) was used for extraction. The organic layers were combined, concentrated, and purified by preparative plate to obtain 79 (HHT0159-101-1A, 50 mg).

[1162] MS m / z(ESI):608[M+1] +

[1163] 1H NMR (400MHz, DMSO-d6) δ12.98(s,1H),10.44(s,1H),8.09(d,J=7.8Hz,1H),7.81(d, J=8.5Hz,1H),7.50–7.34(m,2H),7.23(d,J=8.6Hz,1H),7.03–6.87(m,2H),6.83(s, 1H),6.55(d,J=8.4Hz,1H),4.00(s,2H),3.84–3.52(m,5H),3.51–3.40(m,6H),3.24 (d,J=8.0Hz,2H),2.25–2.07(m,2H),1.86(q,J=9.9,7.6Hz,3H),1.38–1.14(m,3H).

[1164] Embodiment 80

[1165] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(4-methyl-4,7-diazaspiro[2.5]octan-7-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 80

[1166]

[1167] By adopting the same preparation method as Example 50 and using 4-methyl-4,7-diazaspiro[2.5]octane as raw material, Compound 80 can be obtained.

[1168] MS m / z(ESI):619[M+H] + .

[1169] 1H NMR (400MHz, DMSO-d6) δ12.66(s,1H),10.06(s,1H),8.31(d,J=7.7Hz,1H),7.75(d,J=9.1Hz,1H),7.51(s,1H),7.44–7.34(m,2H),7.30(dd,J=8.7, 1.6Hz,1H),7.20(td,J=9.3,4.7Hz,1H),7.11(tt,J=8.0,3.5Hz,1H),6.19 (dd,J=9.2,2.2Hz,1H),6.06(d,J=2.4Hz,1H),5.10(t,J=8.7Hz,1H),4.99 (t,J=8.2Hz,1H),4.91–4.83(m,0H),4.73(dt,J=14.8,7.2Hz,1H),3.78( d,J=11.7Hz,3H),3.72–3.60(m,1H),3.47(t,J=10.9Hz,2H),3.24(t,J=5. 4Hz,2H),3.05(s,2H),2.89(t,J=5.2Hz,2H),2.27(s,3H),1.91(d,J=13.3 Hz,2H),1.39–1.24(m,2H),0.61(q,J=4.4,3.8Hz,2H),0.54–0.43(m,2H).

[1170] Embodiment 81

[1171] Preparation of N-(5-(1-(2,5-difluorophenyl)-2-fluoroethyl)-1H-indazol-3-yl)-4-(3,5-dimethylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 81

[1172]

[1173] Using the same preparation method as Example 50 and using 4-methyl-4,7-diazaspiro[2.5]octane as raw material, Compound 81 can be obtained.

[1174] MS m / z(ESI):607[M+H] + .

[1175] 1H NMR (400MHz, DMSO-d6) δ12.67(s,1H),10.08(s,1H),8.30(d,J=7.7Hz,1H),7.76(d,J=9.0Hz,1H),7.50(s,1H),7.43–7.34(m,2H),7.2 9(dd,J=8.7,1.6Hz,1H),7.19(td,J=9.3,4.7Hz,1H),7.10(ddd,J=8.9,7.5,3.5Hz,1H),6.22(dd,J=9.0,2.3Hz,1H),6.08(d,J=2.3Hz ,1H),5.10(t,J=8.7Hz,0H),4.98(t,J=8.2Hz,1H),4.86(dd,J=9.3,6.4Hz,0H),4.72(dt,J=14.7,7.2Hz,1H),3.82–3.62(m,5H),3.48 (tt,J=11.7,2.3Hz,2H),2.29(t,J=11.4Hz,2H),1.89(dd,J=11.4,6.9Hz,2H),1.32(tq,J=9.3,5.6,4.7Hz,2H),1.06(d,J=6.3Hz,6H).

[1176] Embodiment 82

[1177] Preparation of N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-cyanopyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 82

[1178]

[1179] Using the same preparation method as Example 64, using N-methyl-ethanolamine and 3-cyanopyrrolidine hydrochloride as raw materials, Compound 82 can be obtained.

[1180] MS m / z(ESI):614[M+H] + .

[1181] 1H NMR(400MHz, Methanol-d4)δ7.70(d,J=9.0Hz,1H),7.57(d,J=1.5Hz,1H),7.42–7.37(m,1H),7.22(dd,J=8.7,1.7 Hz,1H),6.85–6.78(m,2H),6.74–6.67(m,1H),6.14(dd,J=9.1,2.4Hz,1H),5.96(d,J=2.4Hz,1H),4.06(s,2H),3.9 2(dt,J=12.0,4.1Hz,2H),3.73–3.63(m,1H),3.63–3.50(m,4H),3.23–3.13(m,1H),3.03(s,3H),2.91–2.79(m,3H ),2.77–2.69(m,2H),2.68–2.58(m,1H),2.26(ddd,J=10.1,7.9,5.5Hz,2H),2.11–1.98(m,3H),1.58–1.46(m,2H).

[1182] Embodiment 83

[1183] Preparation of (R)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-hydroxypyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 83

[1184]

[1185] The first step is the synthesis of tert-butyl 2-nitro-4-(ethyl(2-(p-toluenesulfonyloxy)ethyl)amino)benzoate 83b

[1186] Compound 83a (500 mg, 1.69 mmol) and TEA (338 mg, 3.38 mmol) were dispersed in DCM, and a DCM solution of p-toluenesulfonyl chloride (385 mg, 2.0 mmol) was slowly added at room temperature. The mixture was stirred for 4 hours at room temperature, quenched with water, extracted with DCM, and the organic phase was washed with water, dried over anhydrous sodium sulfate, and concentrated. The crude product was flash purified (PE:EA=1:1) to give 83b (700 mg) as a light yellow solid in a yield of 91%.

[1187] MS m / z(ESI):451[M+H]+.

[1188] Step 2 Synthesis of (R)-tert-butyl 2-nitro-4-(ethyl(2-(3-hydroxypyrrolidin-1-yl)ethyl)amino)benzoate 83c

[1189] Compound 83b (300 mg, 0.66 mmol), (R)-pyrrolidin-3-ol (114 mg, 1.32 mmol) and cesium carbonate (860 mg, 2.64 mmol) were dissolved in DMF (30 mL). The mixture was stirred at room temperature for 12 hours, quenched with water, extracted twice with DCM, the organic phase was washed with water, dried, concentrated, and the crude product was flash purified (DCM: MeOH = 15: 1) to give 83c (200 mg) as a yellow solid. Yield 83%

[1190] MS m / z(ESI):366[M+H]+.

[1191] Step 3 Synthesis of (R)-tert-butyl 2-nitro-4-(ethyl(2-(3-acetoxypyrrolidin-1-yl)ethyl)amino)benzoate 83d

[1192] Compound 83c (150 mg, 0.41 mmol) and TEA (123 mg, 1.23 mmol) were dissolved in DCM, acetyl chloride (48 mg, 0.61 mmol) was added under ice bath, the mixture was stirred at room temperature for 2 hours, quenched by adding water, extracted with DCM, the organic phase was washed with water, dried, concentrated, and the crude product was flash purified (PE:EA=1:1) to obtain 160 mg of compound 83d as a yellow solid. Yield 95%.

[1193] MS m / z(ESI):408[M+H]+.

[1194] Step 4 Synthesis of (R)-tert-butyl 2-amino-4-(ethyl(2-(3-acetoxypyrrolidin-1-yl)ethyl)amino)benzoate 83e

[1195] Compound 83d (160 mg, 0.39 mmol) was dissolved in methanol (10 mL), and zinc powder (127 mg, 1.9 mmol) and saturated ammonium chloride solution (10 mL) were added. The mixture was stirred at room temperature for 2 hours, filtered after the reaction was complete, and the filtrate was extracted with DCM. The organic phase was washed with water, dried, and concentrated to obtain crude compound 83e, which was directly used in the next step.

[1196] MS m / z(ESI):378[M+H]+.

[1197] Step 5 Synthesis of (R)-tert-butyl 2-((tetrahydro-2H-pyran-4-yl)amino)-4-(ethyl(2-(3-acetoxypyrrolidin-1-yl)ethyl)amino)benzoate 83f

[1198] Tetramethylammonium triacetoxyborohydride (512 mg, 1.95 mmol) was slowly added to a solution of compound 83e (150 mg, 0.39 mmol) and tetrahydropyranose ketone (116 mg, 1.16 mmol) in DCM (10 mL) and trifluoroacetic acid (1 mL), and the mixture was stirred at room temperature for 2 hours. Saturated aqueous NaHCO3 solution was added to quench the reaction. The system was extracted with DCM, the organic phase was washed with water, dried over anhydrous sodium sulfate, concentrated, and the crude product 83f was used directly in the next step.

[1199] MS m / z(ESI):462[M+H]+.

[1200] Step 6: Synthesis of 83 g of (R)-tert-butyl 2-(N-(tetrahydro-2H-pyran-4-yl)-trifluoroacetylamino)-4-(ethyl(2-(3-acetoxypyrrolidin-1-yl)ethyl)amino)benzoate

[1201] Compound 83f (150 mg, 0.32 mmol) and TEA (128 mg, 1.28 mmol) were dispersed in DCM, trifluoroacetic anhydride (136 mg, 0.64 mmol) was added dropwise at room temperature, and the mixture was stirred at room temperature for 2 hours. Water was added to quench the reaction. The system was extracted with DCM, the organic phase was washed with water, dried over anhydrous sodium sulfate, and concentrated, and the crude product 83 g was directly used in the next reaction.

[1202] MS m / z(ESI):558[M+H]+.

[1203] Step 7 Synthesis of (R)-2-(N-(tetrahydro-2H-pyran-4-yl)-trifluoroacetylamino)-4-(ethyl(2-(3-acetoxypyrrolidin-1-yl)ethyl)amino)benzoic acid 83h

[1204] Compound 83g (130 mg, 0.23 mmol) was dispersed in DCM (5 mL), trifluoroacetic acid (5 mL) was added dropwise at room temperature, the mixture was stirred at room temperature for 2 hours, concentrated, and the crude compound 83h was directly used in the next reaction.

[1205] MS m / z(ESI):502[M+H]+.

[1206] Step 8 Synthesis of (R)-2-(N-(tetrahydro-2H-pyran-4-yl)-trifluoroacetylamino)-4-(ethyl(2-(3-acetoxypyrrolidin-1-yl)ethyl)amino)benzoyl chloride 83i

[1207] Compound 83h (100 mg, 0.19 mmol) was dispersed in DCM (5 mL), and oxalyl chloride (273 mg, 2.15 mmol) and a drop of DMF were added dropwise at room temperature. The mixture was stirred at room temperature for 2 hours and concentrated. The crude compound 83i was directly used in the next reaction.

[1208] Step 9 Synthesis of (R)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-hydroxypyrrolidin-1-yl)ethyl)amino)-2-(N-(tetrahydro-2H-pyran-4-yl)-trifluoroacetylamino)benzamide 83j

[1209] Compound 5-(3,5-difluorobenzyl)-1H-indazol-3-amine (50 mg, 0.19 mmol) and DIPEA (73 mg, 0.57 mmol) were dissolved in DCM, and a DCM solution of 83i (100 mg, 0.19) was added dropwise under an ice bath. After the addition was complete, the mixture was warmed to room temperature and stirred for 2 hours. Water was added to quench the mixture, and the mixture was extracted with DCM. The organic phase was washed with water, dried over anhydrous sodium sulfate, and concentrated. The crude product was flash purified (DCM: MeOH = 10:1) to obtain 83j (40 mg) as a light yellow solid. The yield was 28%.

[1210] MS m / z(ESI):743[M+H]+.

[1211] Step 10 Synthesis of (R)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-hydroxypyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 83

[1212] Compound 83j (40 mg, 0.053 mmol) was dissolved in methanol (2 mL), and LiOH solution (5 mL) was added. The mixture was stirred at room temperature for 2 hours. After the reaction, DCM was used for extraction. The organic phase was dried and concentrated. The crude product was purified by column chromatography (DCM: MeOH = 10:1) to give compound 83 (5 mg) as a white solid. The yield was 15%.

[1213] MS m / z(ESI):605[M+H]+.

[1214] 1H NMR (400MHz, Methanol-d4) δ7.71(d,J=9.0Hz,1H),7.57(s,1H),7.40(d,J=8.6Hz,1H),7.22(dd,J=8.6,1.6Hz,1H),6 .81(d,J=7.3Hz,2H),6.75–6.66(m,1H),6.14(dd,J=9.1,2.4Hz,1H),5.97(d,J=2.2Hz,1H),4.37(t,J=6.5Hz,1H),4.0 5(s,2H),3.92(dt,J=11.8,4.1Hz,2H),3.68(dt,J=9.5,5.2Hz,1H),3.61–3.51(m,3H),3.03(s,3H),2.95–2.83(m,2H) ,2.81–2.60(m,4H),2.21–2.09(m,1H),2.04(d,J=13.2Hz,2H),1.82–1.71(m,1H),1.53(ddt,J=13.8,9.5,4.9Hz,2H).

[1215] Embodiment 84

[1216] Preparation of (S)-N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(methyl(2-(3-hydroxypyrrolidin-1-yl)ethyl)amino)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide 84

[1217]

[1218] Using the same preparation method as Example 83 and using (S)-3-hydroxypyrrolidine as the starting material, Compound 84 can be obtained.

[1219] MS m / z(ESI):605[M+H] + .

[1220] 1H NMR (400MHz, Methanol-d4) δ7.71(d,J=9.0Hz,1H),7.57(s,1H),7.40(d,J=8.6Hz,1H),7.22(dd,J=8.6,1.6Hz,1H),6 .81(d,J=7.3Hz,2H),6.75–6.66(m,1H),6.14(dd,J=9.1,2.4Hz,1H),5.97(d,J=2.2Hz,1H),4.37(t,J=6.5Hz,1H),4.0 5(s,2H),3.92(dt,J=11.8,4.1Hz,2H),3.68(dt,J=9.5,5.2Hz,1H),3.61–3.51(m,3H),3.03(s,3H),2.95–2.83(m,2H) ,2.81–2.60(m,4H),2.21–2.09(m,1H),2.04(d,J=13.2Hz,2H),1.82–1.71(m,1H),1.53(ddt,J=13.8,9.5,4.9Hz,2H).

[1221] Biological Experiment Example:

[1222] Experiment 1: Determination of in vitro enzymatic activity of compounds

[1223] 1. Experimental Purpose and Methods

[1224] The purpose of this experiment is to test the inhibitory effect of the compounds of the present invention on tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK), and proto-oncogene tyrosine kinase (ROS1). According to the half inhibitory concentration IC 50 The compounds were evaluated for their in vitro activity.

[1225] 2. Experimental Plan

[1226] 2.1 Compound preparation and assay plate preparation

[1227] The test compound was diluted to 50 μM with dimethyl sulfoxide (DMSO), transferred to a 96-well plate and diluted in a 3-fold gradient to 10 concentrations. 100 μL of DMSO was added to two empty wells of the 96-well plate and the plate was marked as the source plate. Then 10 μL of the compound was transferred from the source plate to a new 96-well plate (middle plate). 90 μL of 1x kinase buffer was added to each well of the middle plate. The compounds were mixed in the middle plate for 10 minutes on a shaker. Then 5 μL was transferred from each well of the middle plate to a 384-well plate with 2 replicates for each concentration.

[1228] 2.2 Kinase reaction

[1229] Add 10 microliters (μL) of 2.5x kinase solution (containing tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK) or proto-oncogene tyrosine kinase (ROS1) protein at final concentrations of 2.5 nM, 1.25 nM, and 0.3 nM, respectively) to each well of a 384-well assay plate and incubate at room temperature for 10 minutes (min). Then, 10 μL of 2.5x substrate peptide solution (1x kinase alkaline buffer containing hydroxyfluorescein (FAM)-labeled substrate peptide and adenosine triphosphate (ATP)) was added to each well. At the same time, a control group was set up (the maximum value (Max) well was the DMSO-free group; the minimum value (Min) well was the group without tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK) or proto-oncogene tyrosine kinase (ROS1) protein). After incubation at 28°C for a certain period of time, 25 μL of stop buffer was added to stop the kinase reaction. The amount of substrate and product was determined using the Caliper instrument, and the conversion rate of the product was calculated, and then the inhibition rate was calculated.

[1230] 2.3 Data processing and statistics

[1231] The data were fitted with a curve and the half inhibitory concentration (IC 50 ) value. Inhibition percentage = (maximum (Max) well conversion rate - compound conversion rate per well) / (maximum (Max) well conversion rate - minimum (min) well conversion rate) * 100. "Maximum (max)" represents the dimethyl sulfoxide (DMSO) group without compound; "minimum (min)" represents the group without tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK) or proto-oncogene tyrosine kinase (ROS1) protein.

[1232] 2.4 Test results and conclusions

[1233] The specific IC values ​​of the compounds of the present invention for the activity determination of tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK), and proto-oncogene tyrosine kinase (ROS1) 50 The values ​​are shown in the following Table 1. As can be seen from Table 1, the compounds of the present invention can effectively inhibit the activities of tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK), and proto-oncogene tyrosine kinase (ROS1) at the in vitro biochemical experimental level.

[1234] Table 1. Activity results of compounds against tropomyosin receptor kinase A (TRKA), anaplastic lymphoma kinase (ALK), and proto-oncogene tyrosine kinase (ROS1)

[1235]

[1236]

[1237]

[1238] Experiment 2: Analysis of the inhibitory activity of the compounds of the present invention on proliferation of human anaplastic large cell lymphoma cells (Karpas299 cells)

[1239] 1. Experimental Purpose and Methods

[1240] In this experiment, the cell viability was detected by luminescence method ( The inhibitory activity of the compounds of the present invention on the proliferation of human anaplastic large cell lymphoma cells (Karpas299 cells) was tested by the (CTG) method. The half inhibitory concentration IC 50 Compounds were evaluated for cellular activity.

[1241] 2. Experimental Plan

[1242] 2.1 Cell culture

[1243] Human anaplastic large cell lymphoma cells (Karpas299) were ordered from Nanjing Kebai Biotechnology Co., Ltd. and cultured using Losvi cell culture medium (RPMI 1640) (Corning, 09919004) plus 10% fetal bovine serum (Gibco, 10817010) and 1% penicillin / streptomycin double antibody (Corning, 30002297). After observation under a microscope, it was determined that the cells were in good condition. The cells were transferred to a 15 ml (mL) centrifuge tube and centrifuged at 1000 rpm for 5 minutes. The supernatant was discarded, complete culture medium was added, and the suspension was pipetted into a single cell suspension, which was then cultured in a 37°C, 5% carbon dioxide (CO2) incubator (Thermo Fisher, 3111).

[1244] 2.2 Compound preparation and compound plate preparation

[1245] The compound to be tested was prepared into a 10 millimole per liter (mM) working solution with dimethyl sulfoxide (DMSO). The highest concentration during the test was 1 micromole per liter (μM), and the dilution was performed in a ratio of 1:3, with a total of 9 concentration gradients.

[1246] 2.3 Experimental Procedure

[1247] Day 1: Cell plating

[1248] Take the above cell suspension and count the cells using a cell counter. Use complete culture medium to adjust the cell suspension to the required cell density and then inoculate it into a 96-well plate, 90 microliters (μL) per well to make the cell number 10,000 / well, set up a T0 plate, add 90 microliters (μL) of complete culture medium to the blank control, and place it in a 37°C, 5% carbon dioxide (CO2) incubator (Thermo Fisher, 3111) overnight.

[1249] Day 2: T0 board readings

[1250] CTG analysis was performed after adding 10 μL of medium containing solvent to each well. The CTG reagent (Promega, G7573) was melted and the cell plate was equilibrated to room temperature for 30 minutes. 50 μL of CTG solution was added to each well and shaken on an orbital shaker for 2 minutes to lyse the cells. The cell plate was placed at room temperature for 10 minutes to stabilize the luminescence signal and the luminescence value was read using Envision (purchased from PerkinElmer).

[1251] Day 2: Add the drug to be tested

[1252] Add 10 μL of the diluted test compound solution to a 96-well cell culture plate, set up 3 replicates for each concentration, and set up a negative control group (solvent control group without compound and blank control group without cell but culture medium). Culture the cells for 72 hours (h) at 37°C, 5% CO2, and 95% humidity.

[1253] Day 5: Test plate readings (72 hours after drug treatment)

[1254] Thaw CTG reagent (Promega, G7573) and equilibrate the cell plate to room temperature for 30 minutes, add 50 μL of CTG solution to each well, shake on an orbital shaker for 2 minutes to lyse the cells, place the cell plate at room temperature for 10 minutes to stabilize the luminescence signal, and read the luminescence value using Envision (purchased from PerkinElmer).

[1255] 2.4 Data processing and statistics

[1256] The cell proliferation inhibition rate is expressed by the formula: 1-(V sample -V 空白 ) / (V control -V 空白 )×100%. Where V sample is the reading value of the drug treatment group, V 空白 V is the reading value of the blank control containing no cells but only culture medium, control The reading value of the solvent control group without compound was used. GraphPad Prism 5 software was used to draw the S-shaped dose-survival rate curve using a nonlinear regression model and calculate the half-inhibitory concentration (IC) of the compound on cell proliferation. 50 )value.

[1257] 2.5 Test results and conclusions

[1258] The results showed that the compounds in the examples had a strong inhibitory effect on the proliferation of human anaplastic large cell lymphoma cells (Karpas299). The results are shown in Table 2.

[1259] Experiment 3: Analysis of the proliferation inhibitory activity of the compounds of the present invention on the stable transgenic mouse primary B cell line BaF3 LMNA-NTRK1

[1260] 1. Experimental Purpose and Methods

[1261] The purpose of this experiment is to test the inhibitory activity of the compounds of the present invention on the proliferation of the mouse original B cell stable transgenic line BaF3 LMNA-NTRK1. According to the half inhibitory concentration IC 50 Compounds were evaluated for cellular activity.

[1262] 2. Experimental Plan

[1263] 2.1 Cell culture

[1264] The mouse primary B cell stable strain BaF3 LMNA-NTRK1 was ordered from Kangyuan Broad Biotechnology (Beijing) Co., Ltd. and cultured using Losvi cell culture medium (RPMI 1640) (Corning, 09919004) plus 10% fetal bovine serum (Gibco, 10817010) and 1% penicillin / streptomycin double antibody (Corning, 30002297). After observation under a microscope, it was determined that the cells were in good condition. The cells were transferred to a 15 ml (mL) centrifuge tube and centrifuged at 1000 rpm for 5 minutes. The supernatant was discarded, complete culture medium was added, and the suspension was made into a single cell suspension by pipetting. The suspension was then cultured in a 37°C, 5% carbon dioxide (CO2) incubator (Thermo Fisher, 3111).

[1265] 2.2 Compound preparation and compound plate preparation

[1266] The compound to be tested was prepared into a 10 millimolar (mM) working solution with dimethyl sulfoxide (DMSO). The highest concentration during the test was 1 micromole (μM), and the dilution was performed at 1:3, with a total of 9 concentration gradients.

[1267] 2.3 Experimental Procedure

[1268] Day 1: Cell plating

[1269] Take the above cell suspension and count the cells using a cell counter. Use complete culture medium to adjust the cell suspension to the required cell density and then inoculate it into a 96-well plate, 90 microliters (μL) per well to make the cell number 10,000 / well, set up a T0 plate, add 90 microliters (μL) of complete culture medium to the blank control, and place it in a 37°C, 5% carbon dioxide (CO2) incubator (Thermo Fisher, 3111) overnight.

[1270] Day 2: T0 board readings

[1271] CTG analysis was performed after adding 10 μL of medium containing solvent to each well. The CTG reagent (Promega, G7573) was melted and the cell plate was equilibrated to room temperature for 30 minutes. 50 μL of CTG solution was added to each well and shaken on an orbital shaker for 2 minutes to lyse the cells. The cell plate was placed at room temperature for 10 minutes to stabilize the luminescence signal and the luminescence value was read using Envision (purchased from PerkinElmer).

[1272] Day 2: Add the drug to be tested

[1273] 10 μL of the diluted test compound solution was added to a 96-well cell culture plate, with 3 replicates for each concentration and a negative control group (solvent control group without compound and blank control group without cells but culture medium) set up. Cell culture was performed for 72 hours (h) at 37°C, 5% CO2, and 95% humidity.

[1274] Day 5: Test plate readings (72 hours after drug treatment)

[1275] Thaw CTG reagent (Promega, G7573) and equilibrate the cell plate to room temperature for 30 minutes, add 50 μL of CTG solution to each well, shake on an orbital shaker for 2 minutes to lyse the cells, place the cell plate at room temperature for 10 minutes to stabilize the luminescence signal, and read the luminescence value using Envision (purchased from PerkinElmer).

[1276] 2.4 Data processing and statistics

[1277] The cell proliferation inhibition rate is expressed by the formula: 1-(V sample -V 空白 ) / (V control -V 空白 )×100%. Where V sample is the reading value of the drug treatment group, V 空白 V is the reading value of the blank control containing no cells but only culture medium, control The reading value of the solvent control group without compound was used. GraphPad Prism 5 software was used to draw the S-shaped dose-survival rate curve using a nonlinear regression model and calculate the half-inhibitory concentration (IC) of the compound on cell proliferation. 50 )value.

[1278] 2.5 Test results and conclusions

[1279] The results showed that the compounds of the examples had a strong inhibitory effect on the proliferation of the mouse primary B cell stable transgenic line BaF3 LMNA-NTRK1. The results are shown in Table 2.

[1280] Experiment 4: Analysis of the inhibitory activity of the compounds of the present invention on the proliferation of the mouse primary B cell stable transgenic line BaF3 SLC34A2-ROS1

[1281] 1. Experimental Purpose and Methods

[1282] The purpose of this experiment is to test the inhibitory activity of the compounds of the present invention on the proliferation of the mouse original B cell stable transgenic line BaF3 SLC34A2-ROS1. According to the half inhibitory concentration IC 50 Compounds were evaluated for cellular activity.

[1283] 2. Experimental Plan

[1284] 2.1 Cell culture

[1285] The mouse original B cell stable cell line BaF3 SLC34A2-ROS1 stable cell line was ordered from Kangyuan Broad Biotechnology (Beijing) Co., Ltd. and cultured using Losvi cell culture medium (RPMI 1640) (Corning, 09919004) plus 10% fetal bovine serum (Gibco, 10817010) and 1% penicillin / streptomycin double antibody (Corning, 30002297). After observation under a microscope, it was determined that the cells were in good condition. The cells were transferred to a 15 ml (mL) centrifuge tube and centrifuged at 1000 rpm for 5 minutes. The supernatant was discarded, complete culture medium was added, and the suspension was made into a single cell suspension by pipetting. The suspension was then cultured in a 37°C, 5% carbon dioxide (CO2) incubator (Thermo Fisher, 3111).

[1286] 2.2 Compound preparation and compound plate preparation

[1287] The compound to be tested was prepared into a 10 millimolar (mM) working solution with dimethyl sulfoxide (DMSO). The highest concentration during the test was 1 micromole (μM), and the dilution was performed at 1:3, with a total of 9 concentration gradients.

[1288] 2.3 Experimental Procedure

[1289] Day 1: Cell plating

[1290] Take the above cell suspension and count the cells using a cell counter. Use complete culture medium to adjust the cell suspension to the required cell density and then inoculate it into a 96-well plate, 90 microliters (μL) per well to make the cell number 10,000 / well, set up a T0 plate, add 90 microliters (μL) of complete culture medium to the blank control, and place it in a 37°C, 5% CO2 incubator (Thermo Fisher, 3111) overnight.

[1291] Day 2: T0 board readings

[1292] CTG analysis was performed after adding 10 μL of medium containing solvent to each well. The CTG reagent (Promega, G7573) was melted and the cell plate was equilibrated to room temperature for 30 minutes. 50 μL of CTG solution was added to each well and shaken on an orbital shaker for 2 minutes to lyse the cells. The cell plate was placed at room temperature for 10 minutes to stabilize the luminescence signal and the luminescence value was read using Envision (purchased from PerkinElmer).

[1293] Day 2: Add the drug to be tested

[1294] 10 μL of the diluted test compound solution was added to a 96-well cell culture plate, with 3 replicates for each concentration and a negative control group (solvent control group without compound and blank control group without cells but culture medium) set up. Cell culture was performed for 72 hours (h) at 37°C, 5% CO2, and 95% humidity.

[1295] Day 5: Test plate readings (72 hours after drug treatment)

[1296] Thaw CTG reagent (Promega, G7573) and equilibrate the cell plate to room temperature for 30 minutes, add 50 μL of CTG solution to each well, shake on an orbital shaker for 2 minutes to lyse the cells, place the cell plate at room temperature for 10 minutes to stabilize the luminescence signal, and read the luminescence value using Envision (purchased from PerkinElmer).

[1297] 2.4 Data processing and statistics

[1298] The cell proliferation inhibition rate is expressed by the formula: 1-(V sample -V 空白 ) / (V control -V 空白 )×100%. Where V sample is the reading value of the drug treatment group, V 空白 V is the reading value of the blank control containing no cells but only culture medium, control The reading value of the solvent control group without compound was used. GraphPad Prism 5 software was used to draw the S-shaped dose-survival rate curve using a nonlinear regression model and calculate the half-inhibitory concentration (IC) of the compound on cell proliferation. 50 )value.

[1299] 2.5 Test results and conclusions

[1300] The results showed that the compounds of the examples had a strong inhibitory effect on the proliferation of the mouse primary B cell stable line BaF3 SLC34A2-ROS1. The results are shown in Table 2.

[1301] Table 2. Inhibitory effects of compounds on cell proliferation

[1302]

[1303]

[1304]

[1305]

[1306] Experiment 5: Pharmacokinetics of the compounds of the present invention in rats

[1307] 1. Summary

[1308] Male Sprague Dawley (SD) rats weighing 200-300 g and aged for 8 weeks were used as experimental animals. Liquid chromatography-mass spectrometry / mass spectrometry (LC / MS / MS) was used to determine the drug concentrations in plasma at different times after intravenous and oral administration of the compound of Example 2, the compound of Example 4, the compound of Example 6, the compound of Example 9, the compound of Example 10, the compound of Example 14, the compound of Example 16, the compound of Example 17, the compound of Example 18, the compound of Example 20, the compound of Example 21, the compound of Example 23, the compound of Example 24, the compound of Example 29, the compound of Example 31, the compound of Example 32, the compound of Example 33, the compound of Example 35, the compound of Example 37, the compound of Example 38, the compound of Example 39, the compound of Example 40, the compound of Example 42, the compound of Example 44, the compound of Example 45, the compound of Example 54, the compound of Example 55, the compound of Example 59, the compound of Example 61, the compound of Example 67, the compound of Example 70, the compound of Example 73 and the compound of Example 84, and the relevant pharmacokinetic parameters were calculated. The pharmacokinetic behavior of the compound of the present invention in rats was studied to evaluate its pharmacokinetic characteristics.

[1309] 2. Experimental Plan

[1310] 2.1 Experimental compounds

[1311] Determine the compound of Example 2, the compound of Example 4, the compound of Example 6, the compound of Example 9, the compound of Example 10, the compound of Example 14, the compound of Example 16, the compound of Example 17, the compound of Example 18, the compound of Example 20, the compound of Example 21, the compound of Example 23, the compound of Example 24, the compound of Example 29, the compound of Example 31, the compound of Example 32, the compound of Example 33, the compound of Example 35, the compound of Example 37, the compound of Example 38, the compound of Example 39, the compound of Example 40, the compound of Example 42, the compound of Example 44, the compound of Example 45, the compound of Example 54, the compound of Example 55, the compound of Example 59, the compound of Example 61, the compound of Example 67, the compound of Example 70, the compound of Example 73, and the compound of Example 84.

[1312] 2.2 Preparation of compounds

[1313] A certain amount of compound was weighed and dissolved in physiological saline with 10% Tween 80 for intravenous administration or in water with 0.5% methylcellulose for oral administration to prepare a uniform solution.

[1314] 2.3 Blood collection time and sample processing

[1315] The above compound was administered intravenously and orally to rats at a dose of 10 mg / kg. Blood was collected through the eye socket at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours after administration. 0.2 ml (mL) of blood was collected each time and placed in an EDTA-2K anticoagulant tube. The plasma was separated by centrifugation at 4°C, 6000 rpm for 10 minutes and stored at -80°C.

[1316] 2.4 Experimental results and conclusions

[1317] The pharmacokinetic parameters of the compound of the present invention after administration are shown in the following Table 3. As shown in Table 3, the compound of the present invention has good metabolic characteristics and good bioavailability.

[1318] Table 3. Pharmacokinetic parameters of SD rats after intravenous and oral administration of the example compounds

[1319]

[1320]

[1321] Experiment 6. Pharmacodynamics of compounds of Examples 4, 6, 17, 20, 21, 23, 24, 29, 37, 39, 40, 45, 69, and 82 of the present invention in a mouse transplant tumor model of human anaplastic large cell lymphoma cells (Karpas299 cells)

[1322] 1. Summary

[1323] Using 8-week-old female severe combined immunodeficient (CB-17SCID) mice weighing 16g-18g as experimental animals, the drug efficacy of Example 4, Example 6, Example 17, Example 20, Example 21, Example 23, Example 24, Example 29, Example 37, Example 39, Example 40, Example 45, Example 69, and Example 82 on mouse tumors after oral administration to transplanted tumor mice was determined. The effects of the inventive compounds on tumor growth were explored.

[1324] 2. Experimental Plan

[1325] 2.1 Experimental compounds

[1326] The compound of Example 4, the compound of Example 6, the compound of Example 17, the compound of Example 20, the compound of Example 21, the compound of Example 23, the compound of Example 24, the compound of Example 29, the compound of Example 37, the compound of Example 39, the compound of Example 40, the compound of Example 45, the compound of Example 69, and the compound of Example 82.

[1327] 2.2 Preparation of compounds

[1328] A certain amount of the compound was weighed and dissolved in 10% Tween 80 physiological saline solution to prepare a uniform solution.

[1329] 2.3 Cell culture

[1330] Human anaplastic large cell lymphoma cells (Karpas299 cells) were ordered from Nanjing Kebai Biotechnology Co., Ltd. and cultured using Losvi cell culture medium (RPMI 1640) (Corning, 09919004) plus 10% fetal bovine serum (Gibco, 10817010) and 1% penicillin / streptomycin double antibody (Corning, 30002297). After observation under a microscope, it was determined that the cells were in good condition. The cells were transferred to a 15 ml (mL) centrifuge tube and centrifuged at 1000 rpm for 5 minutes. The supernatant was discarded, complete culture medium was added, and the suspension was pipetted into a single cell suspension, which was then cultured in a 37°C, 5% carbon dioxide (CO2) incubator (Thermo Fisher, 3111).

[1331] 2.4 Experimental Procedure

[1332] Under sterile conditions, human anaplastic large cell lymphoma cells (Karpas299 cells) in the logarithmic growth phase were mixed with Matrigel (Corning, 8274014) and then transplanted subcutaneously on the right side of the back of female severe combined immunodeficient (CB-17SCID) mice. Each mouse was inoculated with 3×10 6 After inoculation, the tumor volume grew to 150 cubic millimeters (mm 3 ), the mice were randomly divided into 15 groups, 6 mice in each group for in vivo efficacy experiments, and the negative control group was given an equal amount of solvent. The specific design is shown in Table 4.

[1333] Table 4. In vivo efficacy test plan of compounds

[1334]

[1335] 2.5 Experimental results and conclusions

[1336] The mouse tumor volume was calculated according to the formula = length × width × width / 2 (mm 3 ), the formula of tumor inhibition rate = 1-(tumor volume of the administration group at the end point - tumor volume of the administration group when grouped) / (tumor volume of the control group at the end point - tumor volume of the control group when grouped) × 100%, the tumor growth inhibition rates of Example compounds 4, 6, 17, 20, 21, 23, 24, 29, 37, 39, 40, 45, 69, and 82 at a dosage concentration of 10 mg / kg were 96.25%, 99.59%, 96.94%, 97.21%, respectively. %, 98.49%, 95.18%, 95.35%, 96.67%, 96.49%, 98.11%, 95.66%, 97.23%, 96.41%, 97.53%; indicating that Example Compounds 4, 6, 17, 20, 21, 23, 24, 29, 37, 39, 40, 45, 69, and 82 of the present invention have a strong tumor growth inhibitory effect in the human anaplastic large cell lymphoma cell (Karpas299 cell) xenograft model.

[1337] Experiment 7: Pharmacodynamics of compounds of Examples 2, 9, 10, 14, 16, 18, 29, 33, 42, 45, 55, 59, 72, and 80 of the present invention in the mouse original B cell stable transgenic strain BaF3 LMNA-NTRK1 mouse transplant tumor model

[1338] 1. Summary

[1339] Female severe combined immunodeficient (CB-17SCID) mice aged 8 weeks weighing 16g-18g were used as experimental animals to measure the efficacy of Example 2, Example 9, Example 10, Example 14, Example 16, Example 18, Example 29, Example 33, Example 42, Example 45, Example 55, Example 59, Example 72, and Example 80 on mouse tumors after oral administration to transplanted tumor mice. The effects of the inventive compounds on tumor growth were explored.

[1340] 2. Experimental Plan

[1341] 2.1 Experimental compounds

[1342] The compound of Example 2, the compound of Example 9, the compound of Example 10, the compound of Example 14, the compound of Example 16, the compound of Example 18, the compound of Example 29, the compound of Example 33, the compound of Example 42, the compound of Example 45, the compound of Example 55, the compound of Example 59, the compound of Example 72, and the compound of Example 80.

[1343] 2.2 Preparation of compounds

[1344] A certain amount of the compound was weighed and dissolved in 10% Tween 80 physiological saline solution to prepare a uniform solution.

[1345] 2.3 Cell culture

[1346] The mouse primary B cell stable strain BaF3 LMNA-NTRK1 was ordered from Kangyuan Broad Biotechnology (Beijing) Co., Ltd. and cultured using Losvi cell culture medium (RPMI 1640) (Corning, 09919004) plus 10% fetal bovine serum (Gibco, 10817010) and 1% penicillin / streptomycin double antibody (Corning, 30002297). After observation under a microscope, it was determined that the cells were in good condition. The cells were transferred to a 15 ml (mL) centrifuge tube and centrifuged at 1000 rpm for 5 minutes. The supernatant was discarded, complete culture medium was added, and the suspension was made into a single cell suspension by pipetting. The suspension was then cultured in a 37°C, 5% carbon dioxide (CO2) incubator (Thermo Fisher, 3111).

[1347] 2.4 Experimental Procedure

[1348] Under sterile conditions, the logarithmic growth phase mouse primary B cell stable strain BaF3 LMNA-NTRK1 was mixed with Matrigel (Corning, 8274014) and then transplanted subcutaneously on the right back of female severe combined immunodeficient (CB-17SCID) mice. Each mouse was inoculated with 5×10 6After inoculation, the tumor volume grew to 150 cubic millimeters (mm 3 ), the mice were randomly divided into 15 groups, 6 mice in each group for in vivo efficacy experiments, and the negative control group was given an equal amount of solvent. The specific design is shown in Table 5.

[1349] Table 5. In vivo efficacy test plan of compounds

[1350]

[1351]

[1352] 2.5 Experimental results and conclusions

[1353] The mouse tumor volume was calculated according to the formula = length × width × width / 2 (mm 3 ), tumor inhibition rate formula = 1-(tumor volume of the administration group at the end point - tumor volume of the administration group when grouped) / (tumor volume of the control group at the end point - tumor volume of the control group when grouped) × 100%, Example compounds 2, 9, 10, 14, 16, 18, 29, 33, 42, 45, 55, 59, 72, 80 at a dosage concentration of 10 mg / kg had tumor growth inhibition rates of 96.25%, 97.49%, 96.34%, 97.17%, respectively. %, 98.23%, 97.35%, 95.25%, 96.46%, 96.20%, 97.42%, 97.23%, 98.36%, 96.34%, 98.06%; indicating that the example compounds 2, 9, 10, 14, 16, 18, 29, 33, 42, 45, 55, 59, 72, and 80 of the present invention have a strong tumor growth inhibitory effect in the mouse original B cell stable line BaF3LMNA-NTRK1 xenograft model.

[1354] Experiment 8. Pharmacodynamics of compounds of Examples 4, 9, 14, 17, 18, 21, 29, 33, 42, 45, 59, 61, 76, and 83 of the present invention in a mouse proto-B cell stable transgenic strain Ba / F3 SLC34A2-ROS1 mouse transplant tumor model

[1355] 1. Summary

[1356] Female severe combined immunodeficient (CB-17SCID) mice aged 8 weeks weighing 16g-18g were used as experimental animals to measure the efficacy of Example 4, Example 9, Example 14, Example 17, Example 18, Example 21, Example 29, Example 33, Example 42, Example 45, Example 59, Example 61, Example 76, and Example 83 on mouse tumors after oral administration to transplanted tumor mice. The effects of the inventive compounds on tumor growth were explored.

[1357] 2. Experimental Plan

[1358] 2.1 Experimental compounds

[1359] The compound of Example 4, the compound of Example 9, the compound of Example 14, the compound of Example 17, the compound of Example 18, the compound of Example 21, the compound of Example 29, the compound of Example 33, the compound of Example 42, the compound of Example 45, the compound of Example 59, the compound of Example 61, the compound of Example 76, and the compound of Example 83.

[1360] 2.2 Preparation of compounds

[1361] A certain amount of the compound was weighed and dissolved in 10% Tween 80 physiological saline solution to prepare a uniform solution.

[1362] 2.3 Cell culture

[1363] Mouse pro-B cell stable strain Ba / F3 SLC34A2-ROS1 was ordered from Kangyuan Broad Biotechnology (Beijing) Co., Ltd. and cultured using Losvi cell culture medium (RPMI 1640) (Corning, 09919004) plus 10% fetal bovine serum (Gibco, 10817010) and 1% penicillin / streptomycin double antibody (Corning, 30002297). After observation under a microscope, it was determined that the cells were in good condition. The cells were transferred to a 15 ml (mL) centrifuge tube and centrifuged at 1000 rpm for 5 minutes. The supernatant was discarded, complete culture medium was added, and the suspension was made into a single cell suspension by pipetting. The suspension was then cultured in a 37°C, 5% carbon dioxide (CO2) incubator (Thermo Fisher, 3111).

[1364] 2.4 Experimental Procedure

[1365] Under sterile conditions, the mouse primary B cell stable strain Ba / F3 SLC34A2-ROS1 in the logarithmic growth phase was mixed with Matrigel (Corning, 8274014) and then transplanted subcutaneously on the right side of the back of female severe combined immunodeficient (CB-17SCID) mice. Each mouse was inoculated with 1*106 After inoculation, the tumor volume grew to 150 cubic millimeters (mm 3 ), the mice were randomly divided into 15 groups, 6 mice in each group for in vivo efficacy experiments, and the negative control group was given an equal amount of solvent. The specific design is shown in Table 6.

[1366] Table 6. In vivo efficacy test plan of compounds

[1367]

[1368]

[1369] 2.5 Experimental results and conclusions

[1370] The mouse tumor volume was calculated according to the formula = length × width × width / 2 (mm 3 ), tumor inhibition rate formula = 1-(tumor volume of the administration group at the end point - tumor volume of the administration group when grouped) / (tumor volume of the control group at the end point - tumor volume of the control group when grouped) × 100%, Example compounds 4, 9, 14, 17, 18, 21, 29, 33, 42, 45, 59, 61, 76, 83 at a dosage concentration of 10 mg / kg had tumor growth inhibition rates of 92.13%, 96.72%, 94.36%, 91.17%, respectively. , 97.28%, 96.46%, 94.03%, 96.22%, 95.37%, 95.24%, 97.80%, 96.69%, 97.32%, 96.27%; indicating that the example compounds 4, 9, 14, 17, 18, 21, 29, 33, 42, 45, 59, 61, 76, and 83 of the present invention have a strong tumor growth inhibitory effect in the mouse original B cell stable line Ba / F3SLC34A2-ROS1 xenograft model.

[1371] Experiment 9. Acute toxicity test on rats of compounds of Example 11, Example 15, Example 24, Example 45, Example 59, Example 62, Example 77 and Example 81 of the present invention

[1372] 1. Experimental Purpose and Methods

[1373] The purpose of this experiment is to test the toxic effects of compounds in rats.

[1374] Rats were given a single oral administration of 100 mg / kg, 200 mg / kg and 500 mg / kg of Example 11, Example 15, Example 24, Example 45, Example 59, Example 62, Example 77 and Example 81 of the present invention, and observed for 14 days, and the animal deaths, poisoning reactions, weight changes, diet, appearance, behavior, etc. were recorded. At the end point, the animals were dissected, and the organs were removed for histopathological examination.

[1375] 2. Experimental results and conclusions

[1376] The median lethal dose (LD50) of the compounds of Examples 11, 15, 24, 45, 59, 62, 77 and 81 of the present invention is greater than 500 mg / kg, and the safety is good. Compared with the rats in the control group, the rats in the administration group had no abnormal weight and behavior within 14 days from the date of administration, and the compounds of Examples 11, 15, 24, 45, 59, 64, 70 and 82 of the present invention did not show obvious toxicity.

[1377] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

[1378] References:

[1379] 1.Menichincheri M, Ardini E, Magnaghi P, et al. Discovery of Entrectinib: anew 3-aminoindazole as a potent Anaplastic Lymphoma Kinase (ALK), c-rosOncogene1Kinase (ROS1), and Pan-Tropomyosin Receptor Kinases (Pan-TRKs) inhibitor [J]. Journal of Medicinal Chemistry,2016,59,3392-3408.

Claims

1. A compound represented by general formula (II) and a pharmaceutically acceptable salt thereof, in, Y 1 is carbon, Y 3 Nitrogen, Y 2 For carbon, Y 4 For carbon, Y 5 for carbon; R 0 for R 1 for R 2 for R 3 -NH-C 4-8 Cycloalkyl, -NH-C 3-8 Heterocycloalkyl, -NH-CH2-C 3-8 Heterocycloalkyl, -OC 1-4 alkyl; R 6 is phenyl or 4-6 membered heterocycloalkyl, wherein the phenyl or 4-6 membered heterocycloalkyl is mono- or di-substituted by the following groups: halogen, -OH, -C 1-3 Alkyl, -C(CH3)2-OH or cyano; R 7 -CH2-OH, -CH2-OC 1-3 Alkyl, -CH2-OC(O)-C 1-3 Alkyl, -CH2-F, -CH2-OCH3.

2. The compound and pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R 6 It is phenyl or 4-6 membered heterocycloalkyl, wherein the phenyl is substituted by fluorine, and the 4-6 membered heterocycloalkyl is substituted by -OH or -C(CH3)2-OH.

3. The compound and pharmaceutically acceptable salt thereof according to claim 2, characterized in that: R 3 for -OC 1-4 Alkyl, -NH-CH2-C 4-5 Heterocycloalkyl; T 1 is -O, -S, -NH; R 6 for 4. A compound and a pharmaceutically acceptable salt thereof, characterized in that: The compound is: 。 5. A pharmaceutical composition, characterized in that The invention comprises a therapeutically effective amount of the compound according to any one of claims 1 to 4 and a pharmaceutically acceptable salt thereof, and optionally one or more pharmaceutically acceptable carriers and / or diluents.

6. Use of the compound according to any one of claims 1 to 4 and its pharmaceutically acceptable salt or the pharmaceutical composition according to claim 5 in the preparation of a medicament for preventing and / or treating cancer mediated by TRK, ALK or ROS1.

7. The use according to claim 6, characterized in that The cancer is lung cancer, bone marrow cancer, squamous cell carcinoma, thyroid follicle cancer, prostate cancer, breast cancer, bladder cancer, lung cancer, stomach cancer, colorectal cancer, lymphoma or leukemia.

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