N, 2, 3-trimethyl-N-condensed aryl-2H-indazole-6-amine compound as well as preparation method and application thereof

Abstract

The invention belongs to the field of medical biology, and discloses an N, 2, 3-trimethyl-N-fused aryl-2H-indazole-6-amine compound with a structural formula as shown in a formula I in the specification. The N, 2, 3-trimethyl-N-fused aryl-2H-indazole-6-amine compound disclosed by the invention can be used for effectively inhibiting in-vitro proliferation of a plurality of tumor cells; the action mechanism is similar to that of colchicine, and aggregation of microtubules can be inhibited; the potential development and application values are realized.

Description

Technical Field

[0001] The present invention belongs to the field of chemical medicine, and particularly relates to an N,2,3-trimethyl-N-fused aryl-2H-indazole-6-amine compound, a preparation method thereof and uses thereof. Background Art

[0002] Microtubules are composed of α-tubulin and β-tubulin heterodimers and are the main components of the cytoskeleton, playing an important role in maintaining cell morphology, cell division, signal transduction, material transportation and other processes. Compounds acting on the colchicine binding site can inhibit the further assembly of tubulin dimers into microtubules, thereby directly affecting cell mitosis, arresting cells at the G2 / M phase, and having good proliferation inhibitory activity against a variety of tumor drug-resistant cells. In recent years, it has also been found that compounds acting on the colchicine binding site can disrupt the formed tumor blood vessels and have good clinical effects when combined with targeted tumor angiogenesis inhibitors, so they have attracted the attention of researchers. At the same time, colchicine shows anti-inflammatory effects clinically and is used for the secondary prevention of acute gout and cardiovascular diseases.

[0003] In the literature Journal of Medicinal Chemistry 2008,51,4632~4640, the compound pazopanib (A) was reported. It is a vascular endothelial growth factor receptor (VEGFR) inhibitor and is clinically used for the treatment of advanced renal cell carcinoma, soft tissue sarcoma, epithelial ovarian cancer and non-small cell lung cancer. The report of this compound has attracted attention to related structures, especially the indazole ring in drug design.

[0004] In the literature Chinese Chemical Letters 2014,25,989~994, the compound B was reported. By introducing a 5- or 6-membered cycloalkane at the 4th and 5th positions of the pyrimidine ring, it was found that the compound lost the inhibitory activity against tumor angiogenesis-related kinase receptors such as VEGFR, FGFR, PDGFR, and c-KIT.

[0005] In the literature Journal of Medicinal Chemistry 2009,52,2341~2351, the compound C (Verubulin) was reported. It has significant proliferation inhibitory activity against a variety of tumor cells in vitro, and the IC 50 value is between 1 and 10 nM. However, due to the relatively large toxicity of compound C in phase I and phase II clinical trials, its further development is limited. However, as the first discovered microtubule aggregation inhibitor containing a quinazoline structure, it has attracted extensive research in the industry.

[0006] Compound D was reported in the literature ChemMedChem 2014, 9(4), 847 - 854. It was obtained by replacing the 4 - methoxybenzene ring in compound C with a 6 - substituted - N - methylindole ring and showed strong activities similar to those of compound C in various in vitro tumor cell proliferation inhibition activity tests. The literature Journal of Medicinal Chemistry 2021, 64, 12964 - 12977 obtained compound E by introducing a methyl group at the 2 - position of the indole ring in compound D. The proliferation inhibition activities against MCF - 7 and doxorubicin - resistant MCF - 7 / ADR breast cancer cells were comparable, and the IC 50 values were both 0.9 nM. In the literature European Journal of Medicinal Chemistry 2021, 223, 113656, by further modifying compound E, replacing the 2 - methylquinazoline ring with a 2 - methylquinoline ring and introducing a hydroxymethyl group at the 2 - position of the indole ring to obtain compound F, the proliferation inhibition activity (IC 50 value) against HCT116 tumor cells in vitro reached 0.08 nanomolar.

[0007] Compound G was reported in the literature European Journal of Medicinal Chemistry 2019, 168, 176 - 188. It was obtained by replacing the 2 - methylquinazoline ring and 4 - methoxybenzene ring in compound C with a 2 - methylquinoline ring and a 4 - substituted - N - methylcarbazole ring respectively, and showed sub - nanomolar level activities in various in vitro tumor cell proliferation inhibition activity tests. Especially, the IC 50 value for inhibiting the proliferation of the HCT116 tumor cell line reached 0.07 nanomolar.

[0008] The New England Journal of Medicine 2020, 383, 1838 - 1847 reported that in a randomized trial involving patients with chronic coronary artery disease, the risk of cardiovascular events in patients taking 0.5 mg of colchicine once a day was significantly lower than that in patients taking placebo.

[0009] The structural formulas of the above - mentioned compounds A - G are as follows:

[0010]

[0011] Currently, cancer is the disease that causes the largest number of deaths among all diseases. The drugs used in clinical practice have characteristics such as large adverse reactions and easy development of drug resistance. There is an urgent need to develop new drugs that are safe, effective, low - toxic and can overcome drug resistance.

[0012] Therefore, those skilled in the art are committed to developing a class of N,2,3-trimethyl-N-fused aryl-2H-indazole-6-amine compounds, their preparation methods and uses, which are of great significance for enhancing drug specificity, effectiveness, reducing toxic side effects and preventing drug resistance. At the same time, the compounds in the present invention bind to the same position on tubulin as colchicine and may also have potential anti-inflammatory activity. Summary of the Invention

[0013] Aiming at the deficiencies existing in the above-mentioned prior art, the present invention provides a compound with the structure of formula I, its preparation method and application. This class of compounds is a novel N,2,3-trimethyl-N-fused aryl-2H-indazole-6-amine small molecule tubulin aggregation inhibitor. Further experiments have confirmed that it has significant inhibitory effects on the proliferation of human non-small cell lung cancer A549 cells, human liver cancer Huh-7 cells, human cervical cancer Hela cells, human glioma U251 cells, human liver cancer HepG-2 cells, etc. No relevant activity data have been reported. The present invention is of great significance for enhancing drug specificity, effectiveness, reducing toxic side effects and preventing drug resistance. At the same time, the compounds discovered in the present invention bind to the same position on tubulin as colchicine and may also have potential anti-inflammatory activity.

[0014] To achieve the above object, the present invention adopts the following technical solutions:

[0015] In the first aspect of the present invention, there is provided a class of compounds or their pharmaceutically acceptable salts, which have the structure shown in formula I:

[0016]

[0017] Wherein,

[0018] X is N or CH;

[0019] R 1 is H, halogen, cyano, carboxyl, C 1～6 alkyl, halo C 1～6 alkyl, C 3～12 cycloalkyl, halo C 3～12 cycloalkyl, hydroxy, hydroxy-substituted C 1～6 alkyl, hydroxy-substituted C 3～12 cycloalkyl, C 1～6 alkoxy, C 3～12 cycloalkoxy, amino, C 1～6 alkylamino, C 3～12 cycloalkylamino, C 2～6 azacycloalkylamino, halogen-substituted C 2～6 azacycloalkylamino, hydroxy-substituted C 1～6 alkylamino, hydroxy-substituted C 3～12 cycloalkylamino, hydroxy-substituted C2～6 Azacycloalkylamino, amino-substituted C 1～6 Alkylamino, amino-substituted C 3～12 Cycloalkylamino, amino-substituted C 2～6 Azacycloalkylamino, hydroxy-substituted C 1～6 Dialkylamino, amino-substituted C 1～6 Dialkylamino, heterocycloalkyl, aryl, arylamino, heteroaryl or heteroarylamino, wherein the aryl or heteroaryl ring optionally contains 0 to 2 N atoms, or simultaneously contains 0 to 1 nitrogen atom and oxygen atom, or simultaneously contains 0 to 1 nitrogen atom and sulfur atom; the aryl or heteroaryl is optionally substituted by 1 or 2 R 5 substituents;

[0020] R 2 、R 3 、R 4 are each independently H, C 1～4 alkyl, C 1～4 alkoxy, halogen, cyano, hydroxy, C 1～4 alkylamino, C 1～4 dialkylamino, C 3～6 cycloalkyl or C 3～6 cycloalkylamino;

[0021] R 5 is independently H, C 1～4 alkyl, C 1～4 alkoxy, halogen, cyano, hydroxy, C 1～4 alkylamino, C 1～4 dialkylamino, N-hydroxycarbamoyl-substituted C 0～5 alkyl, N-hydroxycarbamoyl-substituted C 0～5 alkoxy, N-hydroxycarbamoyl-substituted C 0～5 alkylamino, hydrazinocarbonyl-substituted C 0～5 alkyl, hydrazinocarbonyl-substituted C 0～5 alkoxy, hydrazinocarbonyl-substituted C 0～5 alkylamino, C 3～6 cycloalkyl, C 3～6 cycloalkylamino, C 3～6 cyclic amino group containing 1 to 2 heteroatoms, C 3～6 cycloalkylamino group containing 1 to 2 heteroatoms, C 1～6 alkylsulfone, C 1～6 alkylaminomethanesulfonyl or C 1～6 heterocycloalkylaminomethanesulfonyl.

[0022] The second aspect of the present invention relates to a method for preparing the compound of the above general formula I or a pharmaceutically acceptable salt thereof.

[0023] The third aspect of the present invention relates to a pharmaceutical composition comprising at least one of the above-mentioned compounds of general formula I or a pharmaceutically acceptable salt thereof and one or more pharmaceutical carriers or excipients.

[0024] The fourth aspect of the present invention relates to the use of the compound of general formula I or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of a disease or disorder such as a tumor, especially cancer, or for reducing the severity of said disease or disorder, wherein said disease is selected from lung cancer, breast cancer, prostate cancer, gastric cancer, nasopharyngeal cancer, colorectal cancer, ovarian cancer, multiple myeloma, melanoma, glioblastoma multiforme, lymphoma, blood cancer, kidney cancer, liver tumor, sarcoma and thyroid cancer, as well as various other solid tumors.

[0025] The fifth aspect of the present invention relates to a method for treating a disease or disorder such as a tumor, inflammation, which is cancer, or for reducing the severity of said disease or disorder, said method comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of general formula I or a pharmaceutically acceptable salt thereof, wherein said disease is selected from lung cancer, breast cancer, prostate cancer, gastric cancer, nasopharyngeal cancer, colorectal cancer, ovarian cancer, multiple myeloma, melanoma, glioblastoma multiforme, lymphoma, blood cancer, kidney cancer, liver tumor, sarcoma and thyroid cancer, as well as various other solid tumors.

[0026] The sixth aspect of the present invention relates to at least one of the above-mentioned compounds of general formula I or a pharmaceutically acceptable salt thereof, said compound or its pharmaceutically acceptable salt being used for the treatment of a disease or disorder such as a tumor, especially cancer, or for reducing the severity of said disease or disorder, wherein said disease is selected from lung cancer, breast cancer, prostate cancer, gastric cancer, nasopharyngeal cancer, colorectal cancer, ovarian cancer, multiple myeloma, melanoma, glioblastoma multiforme, lymphoma, blood cancer, kidney cancer, liver tumor, sarcoma and thyroid cancer, as well as various other solid tumors.

[0027] Substituent definitions

[0028] As used herein, the term "halogen" means fluorine, chlorine, bromine or iodine. Preferred halogen groups are fluorine, chlorine or bromine.

[0029] As used herein, the term "cyano" means -CN.

[0030] As used herein, the term "carboxyl" means -C(O)OH.

[0031] As used herein, the term "alkyl" refers to a saturated straight-chain or branched-chain monovalent hydrocarbon group having 1 to 6 carbon atoms, preferably having 1 to 6, 1 to 4 or 1 to 3 carbon atoms. Typical examples of "alkyl" include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, tert-pentyl, neopentyl, hexyl, etc.

[0032] The term "cycloalkyl" as used herein means a saturated cyclic hydrocarbon group having 3 to 12 carbon atoms and having a monocyclic or bicyclic or multiple fused rings (including fused and bridged ring systems), preferably having 3 to 10, 3 to 8, 5 to 8, 3 to 6 or 5 to 6 carbon atoms. Typical examples of "cycloalkyl" include, but are not limited to, monocyclic structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 1-methylcyclopropyl, 2-methylcyclopentyl, 2-methylcyclooctyl, etc., bicyclic structures such as bicyclo[2.2.1]heptyl, etc., and polycyclic structures such as adamantyl, etc.

[0033] The term "hydroxy" as used herein means -OH.

[0034] The term "alkoxy" as used herein means the group -OR 6 , where R 6 is selected from alkyl and cycloalkyl as defined herein. Typical examples of "alkoxy" include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, etc.

[0035] The term "amino" as used herein means -NH2.

[0036] The term "alkylamino" as used herein means the group -NHR 6 , where R 6 is selected from alkyl and cycloalkyl as defined herein. Typical examples of "alkylamino" include, but are not limited to, methylamino, ethylamino, propylamino, butylamino, cyclopropylamino, etc.

[0037] The term "azacycloalkylamino" as used herein means the group where n = 1 to 4, and R 7 is selected from alkyl and halogen as defined herein. Typical examples of "azacycloalkylamino" include, but are not limited to, aziridine, 2-methylaziridine, azetidine, 2-methylazetidine, 3-methylazetidine, pyrrolidine, 2-methylpyrrolidine, 3-methylpyrrolidine, 2,5-dimethylpyrrolidine, 3,3-dimethylpyrrolidine, 3,3-difluoropyrrolidine, piperidine, 2-methylpiperidine, 3-methylpiperidine, 4-methylpiperidine, 2,3-dimethylpiperidine, 2,4-dimethylpiperidine, 2,5-dimethylpiperidine, 2,6-dimethylpiperidine, 4,4-difluoropiperidine, etc.

[0038] The term "dialkylamino" as used herein means the group -NR 8 R 9 , where R 8 and R 9 are each independently selected from alkyl and cycloalkyl as defined herein. Typical examples of "dialkylamino" include, but are not limited to, dimethylamino, diethylamino, dipropylamino, dibutylamino, etc.

[0039] As used herein, the term "heterocycloalkyl" means a cycloalkyl as defined herein containing one, two or more heteroatoms independently selected from N, O and S. Typical examples of "heterocycloalkyl" include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperazinyl, N-methylpiperazinyl, thiazinyl, piperidinyl, morpholinyl, and the like.

[0040] As used herein, the term "aryl" means an unsaturated aromatic carbocyclic group of 5 to 14 carbon atoms having a single ring or two or more fused rings. The aryl preferably has 5 to 10, 5 to 8 or 5 to 6 carbon atoms. Typical examples of "aryl" include, but are not limited to, phenyl, naphthyl, anthracenyl, and the like.

[0041] As used herein, the term "heteroaryl" means a heteroaromatic ring group having 5 to 14 ring members, including monocyclic heteroaromatic rings and polycyclic aromatic rings, wherein the monocyclic aromatic ring is fused to one or more other aromatic rings. The heteroaryl has one or two or more heteroatoms selected from O, S or N. Also included within the scope of the term "heteroaryl" as used herein are groups in which the aromatic ring is fused to one or more non-aromatic rings (carbocyclic or heterocyclic), wherein the linking group or point is on the aromatic ring or non-aromatic ring. The heteroaryl preferably has 5 to 10 ring members, more preferably 5 to 6 ring members. Typical examples of "heteroaryl" include, but are not limited to, furanyl, benzofuranyl, imidazolyl, triazolyl, indolyl, tetrazolyl, pyridyl, pteridinyl, pyrimidinyl, triazolyl, quinolinyl, isoquinolinyl, quinazolinyl, quinoxalinyl, and the like.

[0042] As used herein, the term "N-hydroxycarbamoyl-substituted C 0～5 alkyl" means the group -(CH2) n C(O)NHOH, wherein n = 0 to 5.

[0043] As used herein, the term "N-hydroxycarbamoyl-substituted C 0～5 alkoxy" means the group -O(CH2) n C(O)NHOH, wherein n = 0 to 5.

[0044] As used herein, the term "N-hydroxycarbamoyl-substituted C 0～5 alkylamino" means the group -NH(CH2) n C(O)NHOH, wherein n = 0 to 5.

[0045] As used herein, the term "hydrazinylcarbonyl-substituted C 0～5 alkyl" means the group -(CH2) n C(O)NHNH2, wherein n = 0 to 5.

[0046] As used herein, the term "hydrazinylcarbonyl-substituted C0～5 "Alkoxy" means the group -O(CH2) n C(O)NHNH2, where n = 0 to 5.

[0047] As used herein, the term "hydrazinocarbonyl-substituted C 0～5 alkylamine" means the group -NH(CH2) n C(O)NHNH2, where n = 0 to 5.

[0048] As used herein, the term "C 1～6 alkylsulfone" means the group -S(O)2R 10 . Where R 10 is selected from alkyl and cycloalkyl as defined herein. "C 1～6 alkylsulfone" typical examples include, but are not limited to, methylsulfone, ethylsulfone, vinylsulfone, cyanoethylsulfone, etc.

[0049] As used herein, the term "C 1～6 alkylaminomethanesulfonyl" means the group -S(O)2NH R 11 . Where R 11 is selected from alkyl and cycloalkyl as defined herein.

[0050] As used herein, the term "C 1～6 heterocycloalkylaminomethanesulfonyl" means the group -S(O)2NH R 11 . Where R 11 is selected from heterocycloalkyl, aryl or heteroaryl as defined herein.

[0051] The groups defined by each of the above terms herein may also optionally be mono- or polysubstituted by -OH, -NH2, C 1～4 alkyl, C 1～4 alkylamino, C 1～4 alkoxy or halogen.

[0052] When the compound names used herein are inconsistent with the chemical structural formulas, the chemical structural formulas shall prevail.

[0053] According to a preferred embodiment of the present invention, X in formula I is N or C.

[0054] According to a preferred embodiment of the present invention, R in formula I 1 is H, halogen, cyano, carboxyl, C 1～6 alkyl, halo C 1～6 alkyl, C 3～12 cycloalkyl, halo C 3～12 cycloalkyl, hydroxy, hydroxy-substituted C 1～6 alkyl, hydroxy-substituted C 3～12 cycloalkyl, C 1～6 alkoxy, C3～12 Cycloalkyloxy, amino, C 1～6 alkylamino, C 3～12 cycloalkylamino, C 2～6 azacycloalkylamino, halogen-substituted C 2～6 azacycloalkylamino, hydroxy-substituted C 1～6 alkylamino, hydroxy-substituted C 3～12 cycloalkylamino, hydroxy-substituted C 2～6 azacycloalkylamino, amino-substituted C 1～6 alkylamino, amino-substituted C 3～12 cycloalkylamino, amino-substituted C 2～6 azacycloalkylamino, hydroxy-substituted C 1～6 dialkylamino, amino-substituted C 1～6 dialkylamino, heterocycloalkyl, aryl, arylamino, heteroaryl or heteroarylamino, wherein the aryl or heteroaryl ring optionally contains 0 to 2 N atoms, or simultaneously contains 0 to 1 nitrogen atom and oxygen atom, or simultaneously contains 0 to 1 nitrogen atom and sulfur atom; the aryl or heteroaryl is optionally substituted by 1 or 2 R 5 substituents;

[0055] According to a preferred embodiment of the present invention, R 2 , R 3 , R 4 are each independently H, C 1～4 alkyl, C 1～4 alkyloxy, halogen, cyano, hydroxy, C 1～4 alkylamino, C 1～4 dialkylamino, C 3～6 cycloalkyl or C 3～6 cycloalkylamino;

[0056] According to a preferred embodiment of the present invention, R 5 is a substituent on the aromatic ring of an aryl, arylamino, arylbenzyl, heteroaryl, heteroarylamino or heteroarylbenzyl substituted by R 1 , and can independently be H, C 1～4 alkyl, C 1～4 alkyloxy, halogen, cyano, hydroxy, C 1～4 alkylamino, C 1～4 dialkylamino, N-hydroxycarbamoyl-substituted C 0～5 alkyl, N-hydroxycarbamoyl-substituted C 0～5 alkyloxy, N-hydroxycarbamoyl-substituted C 0～5 alkylamino, hydrazinylcarbonyl-substituted C 0～5 alkyl, hydrazinylcarbonyl-substituted C 0～5 alkyloxy, hydrazinylcarbonyl-substituted C 0～5 alkylamino, C 3～6Naphthenyl, C 3～6 Naphthenylamino, C 3～6 Cyclic amino group containing 1 to 2 heteroatoms, C 3～6 Naphthenylamino group containing 1 to 2 heteroatoms, C 1～6 Alkyl sulfone, C 1～6 Alkylaminomethanesulfonyl or C 1～6 Heterocyclic alkylaminomethanesulfonyl.

[0057] According to a preferred embodiment of the present invention, in general formula I,

[0058] X is N or C;

[0059] R 1 is H, halogen, cyano, carboxyl, C 1～6 alkyl, halogenated C 1～6 alkyl, C 3～12 naphthenyl, halogenated C 3～12 naphthenyl, hydroxy, hydroxy-substituted C 1～6 alkyl, hydroxy-substituted C 3～12 naphthenyl, C 1～6 alkoxy, C 3～12 naphthenoxy, amino, C 1～6 alkylamino, C 3～12 naphthenylamino, C 2～6 azacyclic naphthenylamino, halogen-substituted C 2～6 azacyclic naphthenylamino, hydroxy-substituted C 1～6 alkylamino, hydroxy-substituted C 3～12 naphthenylamino, hydroxy-substituted C 2～6 azacyclic naphthenylamino, amino-substituted C 1～6 alkylamino, amino-substituted C 3～12 naphthenylamino, amino-substituted C 2～6 azacyclic naphthenylamino, hydroxy-substituted C 1～6 dialkylamino, amino-substituted C 1～6 dialkylamino, heterocyclic alkyl, aryl, arylamino, heteroaryl or heteroarylamino, wherein the aryl or heteroaryl ring optionally contains 0 to 2 N atoms, or simultaneously contains 0 to 1 nitrogen atom and oxygen atom, or simultaneously contains 0 to 1 nitrogen atom and sulfur atom; the aryl or heteroaryl is optionally substituted by 1 or 2 R 5 substituents;

[0060] R 2 、R 3 、R 4 are each independently H, C 1～4 alkyl, C 1～4 alkoxy, halogen, cyano, hydroxy, C 1～4 alkylamino, C 1～4 dialkylamino, C 3～6Cycloalkyl or C 3～6 Cycloalkylamino;

[0061] R 5 is a substituent on the aromatic ring of an aryl, arylamino, arylbenzyl, heteroaryl, heteroarylamino or heteroarylbenzyl substituted on R 1 and may independently be H, C 1～4 alkyl, C 1～4 alkoxy, halogen, cyano, hydroxy, C 1～4 alkylamino, C 1～4 dialkylamino, N-hydroxycarbamoyl-substituted C 0～5 alkyl, N-hydroxycarbamoyl-substituted C 0～5 alkoxy, N-hydroxycarbamoyl-substituted C 0～5 alkylamino, hydrazinylcarbamoyl-substituted C 0～5 alkyl, hydrazinylcarbamoyl-substituted C 0～5 alkoxy, hydrazinylcarbamoyl-substituted C 0～5 alkylamino, C 3～6 cycloalkyl, C 3～6 cycloalkylamino, C 3～6 cyclic amino group containing 1 to 2 heteroatoms, C 3～6 cycloalkylamino group containing 1 to 2 heteroatoms, C 1～6 alkylsulfone, C 1～6 alkylaminomethanesulfonyl or C 1～6 heterocycloalkylaminomethanesulfonyl.

[0062] According to a preferred embodiment of the present invention, in general formula I,

[0063] R 1 is H, halogen, C 1～3 alkyl, halo C 1～3 alkyl, C 3～6 cycloalkyl, halo C 3～6 cycloalkyl, hydroxy, hydroxy-substituted C 1～3 alkyl, hydroxy-substituted C 3～6 cycloalkyl, C 1～3 alkoxy, C 3～6 cycloalkoxy, amino, C 1～3 alkylamino, C 3～6 cycloalkylamino, C 2～5 azacycloalkylamino, halo-substituted C 2～5 azacycloalkylamino, hydroxy-substituted C 1～3 alkylamino, hydroxy-substituted C 3～6 cycloalkylamino, hydroxy-substituted C 2～5 azacycloalkylamino, amino-substituted C 1～3 alkylamino, amino-substituted C 3～6 cycloalkylamino, amino-substituted C 2～5Azacycloalkylamino, hydroxy-substituted C 1～3 Dialkylamino, amino-substituted C 1～3 Dialkylamino, heterocycloalkyl, aryl, arylamino, heteroaryl or heteroarylamino, wherein the aryl or heteroaryl ring optionally contains 0 to 2 N atoms, or simultaneously contains 0 to 1 nitrogen atom and oxygen atom, or simultaneously contains 0 to 1 nitrogen atom and sulfur atom; the aryl or heteroaryl is optionally substituted by 1 or 2 R 5 Substituted;

[0064] R 2 、R 3 、R 4 Are each independently H, C 1～3 Alkyl, C 1～3 Alkoxy, halogen, cyano, hydroxy, C 1～3 Alkylamino, C 1～3 Dialkylamino, C 3～6 Cycloalkyl or C 3～6 Cycloalkylamino;

[0065] R 5 Is a substituent on the aromatic ring of the aryl, arylamino, arylbenzyl, heteroaryl, heteroarylamino or heteroarylbenzyl substituted by R 1 , and can independently be H, C 1～3 Alkyl, C 1～3 Alkoxy, halogen, cyano, hydroxy, C 1～3 Alkylamino, C 1～3 Dialkylamino, N-hydroxycarbamoyl-substituted C 0～3 Alkyl, N-hydroxycarbamoyl-substituted C 0～3 Alkoxy, N-hydroxycarbamoyl-substituted C 0～3 Alkylamino, hydrazinylcarbonyl-substituted C 0～3 Alkyl, hydrazinylcarbonyl-substituted C 0～3 Alkoxy, hydrazinylcarbonyl-substituted C 0～3 Alkylamino, hydrazinylcarbonyl-substituted C 0～3 Alkylamino, C 3～6 Cycloalkyl, C 3～6 Cycloalkylamino, C 2～5 Azacycloalkylamino, C 1～6 Alkylsulfone, C 1～6 Alkylaminomethanesulfonyl or C 1～6 Heterocycloalkylaminomethanesulfonyl.

[0066] According to a preferred embodiment of the present invention, in the general formula I,

[0067] R 1 Is H, halogen, C 3～6 Cycloalkyl, halo C 3～6 Cycloalkyl, hydroxy-substituted C 1～3Alkyl, hydroxy-substituted C 3～6 Cycloalkyl, amino, C 1～3 Alkylamine, C 3～6 Cycloalkylamine, C 2～5 Azacycloalkylamine, halogen-substituted C 2～5 Azacycloalkylamine, hydroxy-substituted C 1～3 Alkylamino, hydroxy-substituted C 3～6 Cycloalkylamine, hydroxy-substituted C 2～5 Azacycloalkylamine, amino-substituted C 1～3 Alkylamino, amino-substituted C 3～6 Cycloalkylamine, amino-substituted C 2～5 Azacycloalkylamine, hydroxy-substituted C 1～3 Dialkylamino, amino-substituted C 1～3 Dialkylamino, heterocycloalkyl, aryl, arylamine, heteroaryl or heteroarylamine, wherein the aryl or heteroaryl ring optionally contains 0 to 2 N atoms, or simultaneously contains 0 to 1 nitrogen atom and oxygen atom, or simultaneously contains 0 to 1 nitrogen atom and sulfur atom; the aryl or heteroaryl is optionally substituted by 1 or 2 R 5 Substituted;

[0068] R 2 、R 3 、R 4 Are each independently H, C 1～3 Alkyl, C 1～3 Alkoxy, halogen, cyano, hydroxy, C 1～3 Alkylamino, C 1～3 Dialkylamino, C 3～6 Cycloalkyl or C 3～6 Cycloalkylamine;

[0069] R 5 Is the substituent on the aromatic ring of the aryl, arylamine, arylbenzyl, heteroaryl, heteroarylamine or heteroarylbenzyl substituted on R 1 And can independently be H, C 1～3 Alkyl, C 1～3 Alkoxy, halogen, cyano, hydroxy, C 1～3 Alkylamino, C 1～3 Dialkylamino, C 1～6 Alkylsulfone, C 1～6 Alkylaminomethylsulfonyl or C 1～6 Heterocycloalkylaminomethylsulfonyl.

[0070] Preferably, the compounds of general formula I of the present invention are selected from:

[0071]

[0072]

[0073]

[0074] The corresponding chemical name is:

[0075] N,2,3-Trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1)

[0076] N,2,3-Trimethyl-N-(2-methylquinazolin-4-yl)-2H-indazol-6-amine (Compound 2)

[0077] N,2,3-Trimethyl-N-(2-trifluoromethylquinazolin-4-yl)-2H-indazol-6-amine (Compound 3)

[0078] N,2,3-Trimethyl-N-(2-methyl-8-fluoroquinazolin-4-yl)-2H-indazol-6-amine (Compound 4)

[0079] N,2,3-Trimethyl-N-(2-methylquinolin-4-yl)-2H-indazol-6-amine (Compound 5)

[0080] N,2,3-Trimethyl-N-(2-trifluoromethylquinolin-4-yl)-2H-indazol-6-amine (Compound 6)

[0081] N,2,3-Trimethyl-N-(2-trifluoromethyl-8-fluoroquinolin-4-yl)-2H-indazol-6-amine (Compound 7)

[0082] N,2,3-Trimethyl-N-(6,7-dimethoxyquinazolin-4-yl)-2H-indazol-6-amine (Compound 8)

[0083] N,2,3-Trimethyl-N-(quinazolin-4-yl)-2H-indazol-6-amine (Compound 9)

[0084] N,2,3-Trimethyl-N-(2-methylaminoquinazolin-4-yl)-2H-indazol-6-amine (Compound 10)

[0085] N,2,3-Trimethyl-N-(2-cyclopropylaminoquinazolin-4-yl)-2H-indazol-6-amine (Compound 11)

[0086] N,2,3-Trimethyl-N-(2-(3-hydroxypropylamino)quinazolin-4-yl)-2H-indazol-6-amine (Compound 12)

[0087] N,2,3-Trimethyl-N-(2-(trans-4-hydroxycyclohexylamino)quinazolin-4-yl)-2H-indazol-6-amine (Compound 13)

[0088] N,2,3-Trimethyl-N-(2-(4-methylpiperazin-1-yl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 14)

[0089] N,2,3-Trimethyl-N-(2-(4-trifluoromethylpiperidin-1-yl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 15)

[0090] N,2,3-Trimethyl-N-(2-(3,3-difluoropyrrolidin-1-yl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 16)

[0091] N,2,3-Trimethyl-N-(2-(4,4-difluoropiperidin-1-yl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 17)

[0092] N,2,3-Trimethyl-N-(2-(4-cyanoanilino)quinazolin-4-yl)-2H-indazol-6-amine (Compound 18)

[0093] N,2,3-Trimethyl-N-(2-(4-(2-methoxyethoxy)anilino)quinazolin-4-yl)-2H-indazol-6-amine (Compound 19)

[0094] N,2,3-Trimethyl-N-(2-((3-carbamoylsulfonyl-4-methyl)anilino)quinazolin-4-yl)-2H-indazol-6-amine (Compound 20)

[0095] N,2,3-Trimethyl-N-(2-(4-morpholinosulfonanilino)quinazolin-4-yl)-2H-indazol-6-amine (Compound 21)

[0096] N,2,3-Trimethyl-N-(2-(4-cyanophenyl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 22)

[0097] N,2,3-Trimethyl-N-(2-(3-cyanophenyl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 23)

[0098] N,2,3-Trimethyl-N-(2-(3-fluoro-4-cyanophenyl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 24)

[0099] N,2,3-Trimethyl-N-(2-(4-methylsulfonylphenyl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 25)

[0100] N,2,3-Trimethyl-N-(2-(3-furyl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 26)

[0101] N,2,3-Trimethyl-N-(2-(benzofuran-2-yl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 27)

[0102] N,2,3-Trimethyl-N-(2-(2-fluoro-5-pyridyl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 28) The compounds of general formula I of the present invention can be prepared through the following reaction routes:

[0103]

[0104] wherein R 1 、R 2 、R 3 、R 4 and X are as defined in claim 1.

[0105] In the presence of a base or an acid, a compound of general formula II substituted with a chlorine atom is reacted with a compound of general formula III (R 6 is H) 2,3-dimethyl-6-aminoindazole in a solvent, and then a methyl group is introduced onto the secondary amine N atom through a methylation reagent to obtain compounds 1-8 of general formula I; or in the presence of a base or an acid, a compound of general formula II substituted with a chlorine atom is reacted with a compound of general formula III (R 6 is methyl) N,2,3-trimethyl-6-aminoindazole in a solvent to obtain compounds 1-8 of general formula I. Compound 1 is subjected to dehalogenation, or substitution with a fatty amine / aromatic amine, or coupling with an aromatic ring compound to obtain compounds 9-28 of general formula I.

[0106] More specifically,

[0107] Step (1) reaction: In the presence of 2 - 10 equivalents of a base (such as sodium hydride, cesium carbonate, sodium carbonate, sodium bicarbonate, triethylamine, pyridine, N,N - dimethylaminopyridine, N,N - diisopropylethylamine, potassium carbonate, sodium tert - butoxide, cesium carbonate, potassium carbonate) or a catalytic amount to 2 equivalents of an acid (such as hydrochloric acid, sulfuric acid, hydrobromic acid or acetic acid), and optionally in the presence of auxiliary reagents such as cuprous halide, bis(diphenylphosphino)ferrocene palladium dichloride / 1,1'-bis(diphenylphosphino)ferrocene, palladium acetate, X - Phos, etc., a substituted chloro - compound (general formula II) is reacted with 2,3 - dimethyl - 6 - aminoindazole (general formula III, R = H) in a solvent (such as DMF, acetonitrile, ethanol, tert - butanol, acetone, isopropanol, methanol, THF, toluene, 1,4 - dioxane or DMSO) at a temperature below 200 °C (for example, controlling the temperature with an ice / water bath, oil bath or microwave), and the reaction is carried out for 5 minutes to 24 hours to obtain a secondary amine compound. Further, it is dissolved in a solvent (such as DMF, acetonitrile, acetone, THF, toluene, 1,4 - dioxane or DMSO), a base (such as sodium hydride, cesium carbonate, sodium carbonate, sodium bicarbonate, triethylamine, pyridine, potassium carbonate, sodium tert - butoxide, cesium carbonate, potassium carbonate) and a methylation reagent (such as methyl iodide, dimethyl sulfate) are added, and the reaction is carried out at a temperature below 200 °C (for example, controlling the temperature with an ice / water bath, oil bath or microwave) for 5 minutes to 24 hours to obtain compounds 1 - 8 of general formula I;

[0108] Alternatively, in the presence of 2 - 10 equivalents of a base (such as sodium hydride, cesium carbonate, sodium carbonate, sodium bicarbonate, triethylamine, pyridine, N,N - dimethylaminopyridine, N,N - diisopropylethylamine, potassium carbonate, sodium tert - butoxide, cesium carbonate, potassium carbonate) or a catalytic amount to 2 equivalents of an acid (such as hydrochloric acid, sulfuric acid, hydrobromic acid or acetic acid), and optionally in the presence of auxiliary reagents such as cuprous halide, bis(diphenylphosphino)ferrocene palladium dichloride / 1,1'-bis(diphenylphosphino)ferrocene, palladium acetate, X - Phos, etc., a substituted chloro - compound (general formula II) is reacted with N,2,3 - trimethyl - 6 - aminoindazole (general formula III, R = Me) in a solvent (such as DMF, acetonitrile, ethanol, tert - butanol, acetone, isopropanol, methanol, THF, toluene, 1,4 - dioxane or DMSO) at a temperature below 200 °C (for example, controlling the temperature with an ice / water bath, oil bath or microwave), and the reaction is carried out for 5 minutes to 24 hours to obtain compounds 1 - 8 of general formula I.

[0109] Step (2) reaction: Dissolve compound 1 in an organic solvent (such as DMF, acetonitrile, ethanol, tert-butanol, acetone, isopropanol, methanol, THF, toluene, 1,4-dioxane or DMSO), add palladium on carbon in a mass ratio of 10% - 100% or a catalyst containing palladium metal and platinum metal, and under a hydrogen atmosphere or in the presence of ammonium formate, at a temperature below 100 °C (for example, control the temperature with a water bath, oil bath or microwave), react for 5 minutes to 24 hours, and catalytically hydrogenate and reduce to obtain compound 9 of general formula I;

[0110] Alternatively, in the presence of 2 - 10 equivalents of a base (such as cesium carbonate, sodium carbonate, triethylamine, pyridine, N,N-dimethylaminopyridine, N,N-diisopropylethylamine, potassium carbonate, sodium tert-butoxide, cesium carbonate, potassium carbonate) or a catalytic amount to 2 equivalents of an acid (such as hydrochloric acid, sulfuric acid, hydrobromic acid or acetic acid), and optionally in the presence of auxiliary reagents such as cuprous halide, bis(diphenylphosphino)ferrocene dichloropalladium / 1,1'-bis(diphenylphosphino)ferrocene, palladium acetate, X-Phos, etc., react compound 1 with the corresponding alkylamine, arylamine or arylboronic acid in a solvent (such as DMF, acetonitrile, ethanol, tert-butanol, acetone, isopropanol, methanol, THF, toluene, 1,4-dioxane or DMSO) at a temperature below 200 °C (for example, control the temperature with an ice / water bath, oil bath or microwave), react for 5 minutes to 24 hours, to obtain compounds 10 - 28 of general formula I.

[0111] The compounds of the present invention show strong inhibitory activity in various cancer cell tests. As described below, the compounds show inhibitory activity equivalent to or better than that of the positive control drug paclitaxel in the inhibitory activity tests of human non-small cell lung cancer A549 cells, human liver cancer Huh-7 cells, human cervical cancer Hela cells, human glioma U251 cells, and human liver cancer HepG-2 cells. It is particularly noteworthy that some of these compounds also show strong inhibitory activity against drug-resistant KB-VIN cells. Therefore, the series of compounds of the present invention have broad anti-cancer properties, and in-depth research on them is expected to develop new drugs for the treatment of various tumors or cancers.

[0112] The compounds of the present invention can be used either in their own form or in the form of their pharmaceutically acceptable salts or solvates. The pharmaceutically acceptable salts of the compounds of general formula I include salts formed with pharmaceutically acceptable inorganic acids or organic acids, or with pharmaceutically acceptable inorganic bases or organic bases. Examples of suitable acid addition salts include salts formed with hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, perchloric acid, fumaric acid, acetic acid, propionic acid, succinic acid, glycolic acid, formic acid, lactic acid, maleic acid, tartaric acid, citric acid, pamoic acid, malonic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, hydroxynaphthoic acid, hydroiodic acid, malic acid, tannic acid, etc. Examples of suitable base addition salts include salts formed with sodium, lithium, potassium, magnesium, aluminum, calcium, zinc, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucosamine, procaine, etc. When referring to the compounds of the present invention herein, it includes the compounds of general formula I and their pharmaceutically acceptable salts or solvates.

[0113] According to the present invention, the pharmaceutical composition comprises the compound of general formula I of the present invention and a conventional pharmaceutical carrier or excipient. The pharmaceutical composition can be administered by, for example, oral or parenteral routes. The pharmaceutical composition of the present invention can be prepared into various dosage forms by conventional methods in the art, including but not limited to tablets, capsules, solutions, suspensions, granules or injections, etc., and administered by, for example, oral or parenteral routes.

[0114] It should be further noted that the dosage and usage method of the compounds of the present invention depend on many factors, including the age, weight, gender, natural health status, nutritional status, activity intensity of the compound, administration time, metabolic rate, severity of the disease and the subjective judgment of the treating physician. The preferred dosage is between 0.01 and 100 mg / kg body weight / day.

[0115] Advantages of the present invention:

[0116] (1) The N,2,3-trimethyl-N-fused aryl-2H-indazol-6-amine microtubule protein aggregation inhibitors discovered in the present invention have novel structures and are of great significance for enhancing the specificity and effectiveness of drugs, reducing toxic and side effects and preventing drug resistance, etc.

[0117] (2) The N,2,3-trimethyl-N-fused aryl-2H-indazol-6-amine compounds discovered in the present invention are novel microtubule protein aggregation inhibitors, which have strong proliferation inhibitory activity against human non-small cell lung cancer A549 cells, human liver cancer Huh-7 cells, human cervical cancer Hela cells, human glioma U251 cells, and human liver cancer HepG-2 cells. The mechanism of action is similar to that of colchicine, and it can inhibit microtubule protein polymerization, having good potential for development and application. Description of the Drawings

[0118] Figure 1 Results of the in vitro tubulin aggregation inhibition test for the compound of formula I. Detailed implementation mode

[0119] The following examples are used to further illustrate the present invention, but it does not mean that the present invention is limited thereto.

[0120] Method 1: Dissolve 4-chloro-2,5,6,7,8-substituted quinazoline (1 equivalent) or 4-chloro-2,5,6,7,8-substituted quinoline (1 equivalent) and 2,3-dimethyl-6-amino-2H-indazole (1.1 equivalents) in isopropanol, add 1 drop of concentrated hydrochloric acid, and react at room temperature or under reflux for 12 hours. After the reaction is completed, pour the reactant into a small amount of ice water, adjust the pH to 7 with 50 g·L -1 NaHCO3 solution, precipitate the solid, filter by suction and dry to obtain N,2,3-trimethyl-N-(2,5,6,7,8-substituted quinazolin-4-yl)-2H-indazol-6-amine or N,2,3-trimethyl-N-(2,5,6,7,8-substituted quinolin-4-yl)-2H-indazol-6-amine. Dissolve the obtained secondary amine product (1 equivalent) in N,N-dimethylformamide (2 mL·mmol -1 )), cool to 0 °C in an ice bath, add sodium hydride (60% dispersed in mineral oil, 2 - 3 equivalents), stir for 5 min, then add methyl iodide (1.5 - 3 equivalents), stir for 30 minutes, and then warm to room temperature and continue the reaction for 2 hours. Pour the reactant into an appropriate amount of ice water, precipitate the solid, filter by suction and dry. The crude product is separated by silica gel or neutral alumina column chromatography (eluent: gradient elution of ethyl acetate and petroleum ether, 5% - 100% ethyl acetate) to obtain N,2,3-trimethyl-N-(2,5,6,7,8-substituted quinazolin-4-yl)-2H-indazol-6-amine compounds or N,2,3-trimethyl-N-(2,5,6,7,8-substituted quinolin-4-yl)-2H-indazol-6-amine compounds.

[0121] Example 1: N,2,3-Trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1, Method 1)

[0122]

[0123] 2,4-Dichloroquinazoline (3.82 g, 20 mmol), 2,3-dimethyl-6-amino-2H-indazole (3.22 g, 20 mmol), gave 4.85 g of 2,3-dimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine, yield: 75%. 2,3-Dimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (3.23 g, 10 mmol), sodium hydride (60% dispersed in mineral oil, 0.81 g, 20 mmol), methyl iodide (0.94 mL, 12 mmol). The crude product was recrystallized from MeOH / AcOEt (1:10) to give 2.21 g of a pale yellow solid, namely Compound 1, yield: 65%; melting point: 239 - 241 °C. 1 H NMR (600 MHz, CDCl3) δ 7.75 (d, J = 8.4 Hz, 1H), 7.59 (dd, J = 9.0, 0.6 Hz, 1H), 7.55 - 7.50 (m, 1H), 7.47 (d, J = 0.6 Hz, 1H), 7.06 (dd, J = 8.4, 0.6 Hz, 1H), 6.90 (m, 1H), 6.82 (dd, J = 8.4, 1.8 Hz, 1H), 4.12 (s, 3H), 3.70 (s, 3H), 2.64 (s, 3H). HR-ESI-MS m / z 338.117 3 [M+H] + (Calculated value 338.116 7, C 18 H 17 ClN5).

[0124] Example 2: N,2,3-Trimethyl-N-(2-methylquinazolin-4-yl)-2H-indazol-6-amine (Compound 2, Method 1)

[0125]

[0126] 2-Methyl-4-chloroquinazoline (178 mg, 1 mmol), 2,3-dimethyl-6-amino-2H-indazole (162 mg, 1 mmol), gave 245 mg of 2,3-dimethyl-N-(2-methylquinazolin-4-yl)-2H-indazol-6-amine, yield: 81%. 2,3-Dimethyl-N-(2-methylquinazolin-4-yl)-2H-indazol-6-amine (152 mg, 0.5 mmol), sodium hydride (60% dispersed in mineral oil, 40 mg, 1 mmol), methyl iodide (47 μL, 0.75 mmol), gave 101 mg of a yellow solid, namely Compound 2, yield: 65%; melting point: 185 - 186 °C. 11H NMR (600 MHz, CDCl3) δ 7.77 (d, J = 8.4 Hz, 1H), 7.52 (d, J = 9.0 Hz, 1H), 7.51 - 7.47 (m, 1H), 7.41 (d, J = 1.2 Hz, 1H), 7.15 (d, J = 8.4 Hz, 1H), 6.90 - 6.86 (m, 1H), 6.79 (dd, J = 8.4, 1.2 Hz, 1H), 4.09 (s, 3H), 3.69 (s, 3H), 2.76 (s, 3H), 2.61 (s, 3H). HR-ESI-MS m / z 318.171 8 [M+H] + (Calcd. for 318.171 3, C 19 H 20 N5).

[0127] Example 3: N,2,3-Trimethyl-N-(2-trifluoromethylquinazolin-4-yl)-2H-indazol-6-amine (Compound 3, Method 1)

[0128]

[0129] 2-Trifluoromethyl-4-chloro-quinazoline (58 mg, 0.25 mmol), 2,3-dimethyl-6-amino-2H-indazole hydrochloride (49 mg, 0.25 mmol), gave 2,3-dimethyl-N-(2-trifluoromethylquinazolin-4-yl)-2H-indazol-6-amine 76 mg, yield: 85%. 2,3-Dimethyl-N-(2-trifluoromethylquinazolin-4-yl)-2H-indazol-6-amine (66 mg, 0.19 mmol), sodium hydride (60% dispersed in mineral oil, 30 mg, 0.75 mmol), iodomethane (23 μL, 0.38 mmol), gave a yellow solid 55 mg, namely Compound 3, yield: 81%; melting point: 178 - 180 °C. 1 1H NMR (600 MHz, CDCl3) δ 7.94 (d, J = 8.4 Hz, 1H), 7.60 - 7.62 (m, 2H), 7.50 (s, 1H), 7.15 (d, J = 8.4 Hz, 1H), 7.03 (t, J = 8.4 Hz, 1H), 6.84 (dd, J = 9.0, 1.8 Hz, 1H), 4.14 (s, 3H), 3.75 (s, 3H), 2.66 (s, 3H). HR-ESI-MS m / z 372.144 0 [M+H] + (Calcd. for 372.143 1, C 19 H 16 F3N5).

[0130] Example 4: N,2,3-Trimethyl-N-(2-methyl-8-fluoroquinazolin-4-yl)-2H-indazol-6-amine (Compound 4, Method 1)

[0131]

[0132] 2-Methyl-4-chloro-8-fluoroquinazoline (70 mg, 0.36 mmol), 2,3-dimethyl-6-amino-2H-indazole (93 mg, 0.57 mmol), gave 114 mg of 2,3-dimethyl-N-(2-methyl-8-fluoroquinazolin-4-yl)-2H-indazol-6-amine, yield: 99%. 2,3-Dimethyl-N-(2-methyl-8-fluoroquinazolin-4-yl)-2H-indazol-6-amine (93 mg, 0.29 mmol), sodium hydride (60% dispersed in mineral oil, 35 mg, 0.87 mmol), methyl iodide (54 μL, 0.87 mmol), gave 78 mg of a yellow solid, namely Compound 3, yield: 80%; melting point: 87 - 89 °C. 1 H NMR (600 MHz, CDCl3) δ 7.55 (d, J = 9.0 Hz, 1H), 7.42 (s, 1H), 7.23 - 7.18 (m, 1H), 6.89 (d, J = 8.4 Hz, 1H), 6.82 - 6.76 (m, 2H), 4.10 (s, 3H), 3.69 (s, 3H), 2.82 (s, 3H), 2.62 (s, 3H); HR-ESI-MS m / z 336.161 7 [M + H] + (Calculated value 336.161 9, C 19 H 19 FN5).

[0133] Example 5: N,2,3-Trimethyl-N-(2-methylquinolin-4-yl)-2H-indazol-6-amine (Compound 5, Method 1)

[0134]

[0135] 2-Methyl-4-chloroquinoline (180 mg, 1.01 mmol), 2,3-dimethyl-6-amino-2H-indazole hydrochloride (240 mg, 1.21 mmol), gave 2,3-dimethyl-N-(2-methylquinolin-4-yl)-2H-indazol-6-amine 289 mg, yield: 96%. 2,3-Dimethyl-N-(2-chloroquinolin-4-yl)-2H-indazol-6-amine (91 mg, 0.3 mmol), sodium hydride (60% dispersed in mineral oil, 24 mg, 0.6 mmol), iodomethane (28 μL, 0.45 mmol), gave 65 mg of a white solid, namely Compound 5, yield: 68%; melting point: 189-191 °C. 1 H NMR (600 MHz, CDCl3) δ 8.02 (d, J = 5.4 Hz, 1H), 7.79 (d, J = 7.8 Hz, 1H), 7.58 (t, J = 8.4 Hz, 1H), 7.33 (d, J = 9.0 Hz, 1H), 7.25 (t, J = 7.8 Hz, 1H), 7.11 (s, 1H), 6.99 (s, 1H), 6.57 (dd, J = 9.0, 1.2 Hz, 1H), 4.04 (s, 3H), 3.48 (s, 4H), 2.71 (s, 3H), 2.55 (s, 3H). HR-ESI-MS m / z 317.177 2 [M+H] + (Calculated value 317.176 1,

[0136] C 20 H 21 N4).

[0137] Example 6: N,2,3-Trimethyl-N-(2-trifluoromethylquinolin-4-yl)-2H-indazol-6-amine (Compound 6, Method 1)

[0138]

[0139] 2-Trifluoromethyl-4-chloroquinoline (222 mg, 0.96 mmol), 2,3-dimethyl-6-amino-2H-indazole (129 mg, 0.8 mmol), gave 2,3-dimethyl-N-(2-trifluoromethylquinolin-4-yl)-2H-indazol-6-amine 182 mg, yield: 64%. 2,3-Dimethyl-N-(2-trifluoromethylquinolin-4-yl)-2H-indazol-6-amine (53 mg, 0.15 mmol), sodium hydride (60% dispersed in mineral oil, 12 mg, 0.3 mmol), iodomethane (17 μL, 0.23 mmol), gave 47 mg of a pale yellow solid, namely Compound 6, yield: 85%; melting point: 87 - 89 °C. 11H NMR (500 MHz, DMSO) δ 8.04 (d, J = 8.0 Hz, 1H), 7.70 - 7.76 (m, 1H), 7.60 (m, 2H), 7.47 (s, 1H), 7.34 - 7.32 (m, 1H), 7.04 (d, J = 1.5 Hz, 1H), 6.76 (dd, J = 8.5, 1.5 Hz, 1H), 3.97 (s, 3H), 3.56 (s, 3H), 2.56 (s, 3H). HR-ESI-MS m / z [M+H] + 371.1485 (calcd. 371.1478, C 20 H 18 F3N4).

[0140] Example 7: N,2,3-Trimethyl-N-(2-trifluoromethyl-8-fluoroquinolin-4-yl)-2H-indazol-6-amine (Compound 7, Method 1)

[0141]

[0142] At room temperature, 2-trifluoromethyl-4-chloro-8-fluoroquinoline (100 mg, 0.4 mmol), 2,3-dimethyl-6-amino-2H-indazole (78 mg, 0.48 mmol), cesium carbonate (182 mg, 0.56 mmol), palladium acetate (9 mg, 0.04 mmol), X-Phos (15 mg, 0.08 mmol), diatomaceous earth (200 mg), 0.5 mL of tert-butanol and 3 mL of toluene were successively added to a microwave reaction tube. The air was removed under reduced pressure, and then the mixture was heated by microwave to 150 °C for 30 min under closed conditions. A small amount of dichloromethane was added to the reaction mixture, and the insoluble matter was removed by filtration. The filtrate was evaporated to dryness under reduced pressure. The crude product was purified by Flash column chromatography (neutral alumina column, AcOEt:DCM = 0 - 25%) to obtain 112 mg of 2,3-dimethyl-N-(2-trifluoromethyl-8-fluoroquinolin-4-yl)-2H-indazol-6-amine, with a yield of 75%. 2,3-Dimethyl-N-(2-trifluoromethyl-8-fluoroquinolin-4-yl)-2H-indazol-6-amine (72 mg, 0.2 mmol) was dissolved in 3 mL of DMF, and potassium carbonate (83 mg, 0.6 mmol) and methyl iodide (24 μL, 0.4 mmol) were added. The reaction was carried out overnight at room temperature. The reaction mixture was poured into 50 mL of ice water, and the precipitate was separated by filtration, dried. The crude product was purified by Flash column chromatography (neutral alumina column, DCM = 100%) to obtain 59 mg of a yellow solid, which is Compound 7, with a yield of 76%; melting point: 167 - 169 °C. 11H NMR (600 MHz, CDCl3) δ 7.49 (d, J = 9.0 Hz, 1H), 7.44 (d, J = 9.0 Hz, 1H), 7.39 (s, 1H), 7.32 (t, J = 9.0 Hz, 1H), 7.22 - 7.17 (m, 2H), 6.66 (dd, J = 9.0, 1.8 Hz, 1H), 4.09 (s, 3H), 3.58 (s, 3H), 2.59 (s, 3H). HR-ESI-MS m / z 389.139 4 [M + H] + (calcd 389.138 4, C 20 H 17 F4N4).

[0143] Example 8: N,2,3-Trimethyl-N-(6,7-dimethoxyquinazolin-4-yl)-2H-indazol-6-amine (Compound 8, Method 1)

[0144]

[0145] 4-Chloro-6,7-dimethoxy-quinazoline (134 mg, 0.6 mmol) and 2,3-dimethyl-6-amino-2H-indazole (98 mg, 0.6 mmol) were dissolved in 4 mL of i-PrOH, 1 drop of concentrated hydrochloric acid was added, and the mixture was heated to 120 °C by microwave for 30 min. The reaction solution was cooled to room temperature, and insoluble substances were precipitated. The precipitate was filtered by suction and dried to obtain 168 mg of 2,3-dimethyl-N-(6,7-dimethoxyquinazolin-4-yl)-2H-indazol-6-amine hydrochloride, yield: 96%. 2,3-Dimethyl-N-(6,7-dimethoxyquinazolin-4-yl)-2H-indazol-6-amine hydrochloride (70 mg, 0.2 mmol) was dissolved in 3 mL of DMF, cooled to 0 °C in an ice bath, and then sodium hydride (60% dispersed in mineral oil, 24 mg, 0.6 mmol) was added. After stirring for 5 min, methyl iodide (25 μL, 0.4 mmol) was added. After stirring for 30 min, the temperature was raised to room temperature and the reaction was continued for 2 h. The reaction mixture was poured into about 50 mL of ice water, and solid substances were precipitated. The precipitate was filtered by suction and dried. The crude product was separated by flash neutral alumina column chromatography (AcOEt:MeOH = 97:3) to obtain 40 mg of a yellow solid, which was Compound 8, yield: 55%; melting point: 163 - 166 °C. 11H NMR (600 MHz, d-DMSO) δ 8.63 (s, 1H), 7.76 (d, J = 9.0 Hz, 1H), 7.23 (s, 1H), 7.13 (s, 1H), 6.90 (dd, J = 9.0, 1.8 Hz, 1H), 6.42 (s, 1H), 4.02 (s, 3H), 3.85 (s, 3H), 3.56 (s, 3H), 2.96 (s, 3H), 2.60 (s, 3H). HR-ESI-MS m / z 364.178 2 [M+H] + (Calcd. for 364.176 8, C 20 H 22 N5O2).

[0146] Example 9: N,2,3-Trimethyl-N-(quinazolin-4-yl)-2H-indazol-6-amine (Compound 9)

[0147]

[0148] At room temperature, N,2,3-Trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1, 51 mg, 0.15 mmol) was dissolved in a mixed solvent of 16 mL MeOH / THF (1:1 / v:v), then palladium on carbon (50 mg, 10% W / W) and diisopropylethylamine (53 μL, 0.3 mmol) were added, and the mixture was stirred overnight under a hydrogen atmosphere. The palladium on carbon was filtered off through diatomaceous earth, and the filtrate was evaporated to dryness under reduced pressure. The crude product was separated by flash column chromatography on neutral alumina (AcOEt:PE = 0 - 75%) to obtain 45 mg of a gray solid, which is Compound 9, yield: 99%; melting point: 210 - 212 °C; 1 1H NMR (500 MHz, CDCl3) δ (ppm) 8.75 (s, 1H) 7.76 - 7.72 (m, 2H), 7.63 - 7.59 (m, 1H), 7.33 (d, J = 1.0 Hz, 1H), 7.11 - 7.03 (m, 2H), 6.86 (dd, J = 9.0, 1.5 Hz, 1H), 4.02 (s, 3H), 3.59 (s, 3H), 2.61 (s, 3H); HR-ESI-MS m / z 304.157 0 [M+H] + (Calcd. for 304.155 7, C 18 H 18 N5).

[0149] Example 10: N,2,3-Trimethyl-N-(2-methylaminoquinazolin-4-yl)-2H-indazol-6-amine (Compound 10)

[0150]

[0151] Suspend N,2,3-trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1, 51 mg, 0.15 mmol) and methylamine hydrochloride (60 mg, 0.9 mmol) in 3 mL of i-PrOH, and carry out a microwave reaction for 30 min under closed conditions. Pour the reaction solution into about 30 mL of ice water, precipitate a white solid, filter by suction, and dry. The crude product is separated by flash neutral alumina column chromatography (AcOEt:DCM:MeOH = 120:24:1) to obtain 41 mg of a yellow solid, namely Compound 10, yield: 82%; melting point: 281 - 282 °C; 1 H NMR (600 MHz, CDCl3) δ 7.51 (d, J = 9.0 Hz, 1H), 7.45 (d, J = 8.4 Hz, 1H), 7.41 (s, 1H), 7.36 - 7.32 (m, 1H), 7.03 (d, J = 8.4 Hz, 1H), 6.79 (dd, J = 9.0, 1.2 Hz, 1H), 6.59 (t, J = 7.8 Hz, 1H), 5.16 (brs, 1H), 4.09 (s, 3H), 3.61 (s, 3H), 3.13 (d, J = 5.1 Hz, 3H), 2.61 (s, 3H). HR-ESI-MS m / z 333.183 6 [M + H] + (Calcd. 333.182 2, C 19 H 21 N6).

[0152] Example 11: N,2,3-Trimethyl-N-(2-cyclopropylaminoquinazolin-4-yl)-2H-indazol-6-amine (Compound 11)

[0153]

[0154] Dissolve N,2,3-trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1, 68 mg, 0.2 mmol) in 2 mL of N,N-dimethylacetamide (DMAc), add cyclopropylamine (1 mL, in excess), and heat by microwave to 150 °C for 30 min. Pour the reaction mixture into about 50 mL of ice water, extract with ethyl acetate (30 mL * 3), combine the organic phases, wash with saturated brine, add anhydrous sodium sulfate and dry overnight, remove ethyl acetate under reduced pressure, and separate the crude product by flash neutral alumina column chromatography (AcOEt:PE = 0 - 50%) to obtain 51 mg of a yellow solid. Yield: 71%; melting point: 268 - 270 °C; 11H NMR (600 MHz, CDCl3) δ (ppm) 7.52 (d, J = 8.4 Hz, 1H), 7.49 (d, J = 8.4 Hz, 1H), 7.43 (s, 1H), 7.37 - 7.33 (m, 1H), 7.01 (d, J = 8.4 Hz, 2H), 6.80 (dd, J = 9.0, 1.8 Hz, 1H), 6.63 - 6.59 (m, 1H), 4.10 (s, 3H), 3.63 (s, 3H), 2.97 - 2.94 (m, 1H), 2.61 (s, 3H), 0.87 - 0.83 (m, 2H), 0.67 - 0.63 (m, 2H); HR-ESI-MS m / z 359.198 5 [M+H] + (Calcd 359.197 9, C 21 H 23 N6).

[0155] Method 2: Dissolve N,2,3-trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1, 1 equiv) and the corresponding amine (3 - 4 equiv) in N,N-dimethylacetamide (DMAc) or isopropanol, add diisopropylethylamine (1 mL, in excess), and heat under microwave at 100 - 200 °C for 30 min. Pour the reaction mixture into about 50 mL of ice water, extract with AcOEt (30 mL * 3), combine the organic phases, wash with saturated brine, add anhydrous sodium sulfate and dry overnight, remove AcOEt under reduced pressure, and separate the crude product by flash neutral alumina column chromatography (AcOEt:MeOH = 98:2) to obtain the target product.

[0156] Example 12: N,2,3-Trimethyl-N-(2-(3-hydroxypropylamino)quinazolin-4-yl)-2H-indazol-6-amine (Compound 12, Method 2)

[0157]

[0158] N,2,3-Trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1, 101 mg, 0.3 mmol), 3-aminopropanol (68 mg, 0.9 mmol), DMAc (3 mL), gave 73 mg of a yellow solid. Yield: 65%; Melting point: 87 - 89 °C. 1HNMR(600MHz,CDCl3)δ7.51(d,J=9.0Hz,1H),7.43~7.40(m,2H),7.35~7.31(m,1H),7.00(d,J=8.4Hz,1H),6.78(dd,J=9.0,1.8Hz,1H),6.61~6.57(m,1H),4.09(s,3H),3.74~3.68(m,4H),3.58(s,3H),2.61(s,3H),1.82~1.70(m,2H); HR-ESI-MS m / z 377.208 4[M+H] + (Calcd. 377.201 2, C 21 H 25 N6O).

[0159] Example 13: N,2,3-Trimethyl-N-(2-(trans-4-hydroxycyclohexylamino)quinazolin-4-yl)-2H-indazol-6-amine (Compound 13, Method 2)

[0160]

[0161] N,2,3-Trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1, 51 mg, 0.15 mmol), trans-4-aminocyclohexanol (69 mg, 0.6 mmol), 2 mL of DMAc, gave 34 mg of a pale yellow solid. Yield: 55%; Melting point: 137 - 140 °C. 1 H NMR(600MHz,CDCl3)δ(ppm)7.52(d,J=9.0Hz,1H), 7.41(d,J=9.0Hz,2H), 7.33(t,J=7.8Hz,1H), 6.99(d,J=8.4Hz,2H), 6.80(d,J=9.0Hz,1H), 6.59(t,J=7.8Hz,1H), 5.30(s,1H), 4.10(s,3H), 4.02~3.99(m,1H), 3.73~3.69(m,1H), 3.61(s,3H), 2.62(s,3H), 2.26~2.24(m,2H),2.07~2.04(m,2H),1.54~1.47(m,2H),1.40~1.34(m,2H); HR-ESI-MS m / z 417.240 4[M+H] + (Calcd. 417.239 7, C 24 H 29 N6O).

[0162] Example 14: 2-(4-Methylpiperazin-1-yl)-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 14, Method 2)

[0163]

[0164] 2-Chloro-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 1, 51 mg, 0.15 mmol), 4-methylpiperazine (30 mg, 0.3 mmol), 2 mL i-PrOH, gave 42 mg of a pale yellow solid. Yield: 70%; Melting point: 179 - 181 °C. 1 H NMR (600 MHz, CDCl3) δ 7.50 (d, J = 9.0 Hz, 1H), 7.41 (s, 1H), 7.33 (t, J = 7.2 Hz, 1H), 6.99 (d, J = 8.4 Hz, 1H), 6.78 (dd, J = 8.4, 1.8 Hz, 1H), 6.58 (t, J = 7.8 Hz, 1H), 4.05 - 4.09 (m, 7H), 3.61 (s, 3H), 2.60 - 2.62 (m, 7H), 2.42 (s, 3H); HR-ESI-MS m / z 402.240 8 [M+H] + (Calcd 402.240 1, C 23 H 28 N7).

[0165] Method 3: Suspend 2-chloro-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 1, 1 equiv) and the corresponding amine (2 - 4 equiv) in 5 mL of isopropyl alcohol (i-PrOH), add 1 drop of concentrated sulfuric acid, and heat to 120 °C under microwave irradiation for 30 min. Add a small amount of Et3N to the reaction mixture, remove i-PrOH under reduced pressure, and separate the crude product by flash column chromatography on neutral alumina to obtain the target product.

[0166] Example 15: 2-(4-Trifluoromethylpiperidin-1-yl)-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 15, Method 3)

[0167]

[0168] 2-Chloro-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 1, 51 mg, 0.15 mmol), 4-trifluoromethylpiperidine (46 mg, 0.3 mmol), gave 44 mg of a light pink solid. Yield: 65%; Melting point: 191 - 193 °C. 1HNMR(500MHz, DMSO) δ(ppm) 7.71 (d, J = 8.5Hz, 1H), 7.37 - 7.32 (m, 2H), 7.27 (s, 1H), 6.89 - 6.83 (m, 2H), 6.63 - 6.59 (m, 1H), 4.97 (d, J = 13.0Hz, 2H), 4.02 (s, 3H), 3.52 (s, 3H), 2.94 (t, J = 12.0Hz, 2H), 2.60 (s, 3H), 1.92 (d, J = 12.0Hz, 2H), 1.47 (td, J = 12.5, 4.0Hz, 2H). HR-ESI-MS m / z 455.216 9 [M + H] + (Calcd. 455.216 6, C 24 H 26 F3N6).

[0169] Example 16: 2-(3,3-Difluoropyrrolidin-1-yl)-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 16, Method III)

[0170]

[0171] 2-Chloro-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 1, 51 mg, 0.15 mmol), 3,3-difluoropyrrolidine hydrochloride (43 mg, 0.3 mmol), gave 39 mg of a yellow solid. Yield: 64%; Melting point: 195 - 196 °C. 1 H NMR(600MHz, CDCl3) δ(ppm) 7.53 (d, J = 7.2Hz, 1H), 7.42 (s, 1H), 7.36 (brs, 1H), 7.01 (brs, 1H), 6.78 (d, J = 9.0Hz, 1H), 6.61 (brs, 1H), 4.17 - 4.00 (m, 7H), 3.63 (s, 3H), 2.62 (s, 3H), 2.52 - 2.49 (m, 2H); HR-ESI-MS m / z 409.195 1 [M + H] + (Calcd. 409.194 7, C 22 H 23 F2N6).

[0172] Example 17: 2-(3,3-Difluoropiperidin-1-yl)-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 17, Method III)

[0173]

[0174] 2-Chloro-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 1, 68 mg, 0.2 mmol), 4,4-difluoropiperazine (48 mg, 0.4 mmol), gave 58 mg of a yellow solid. Yield: 69%; Melting point: 191 - 192 °C. 1 H NMR (600 MHz, DMSO) δ (ppm) 7.72 (d, J = 9.0 Hz, 1H), 7.39 - 7.34 (m, 2H), 7.28 (s, 1H), 6.90 - 6.85 (m, 2H), 6.65 - 6.61 (m, 1H), 4.03 - 4.00 (m, 7H), 3.53 (s, 3H), 2.61 (s, 3H), 2.09 - 2.02 (m, 4H); HR-ESI-MS m / z 423.211 8 [M+H] + (Calculated value 423.210 3, C 23 H 25 F2N6).

[0175] Example 18: 2-(4-Cyanoanilino)-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 18, Method III)

[0176]

[0177] 2-Chloro-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 1, 52 mg, 0.15 mmol), 4-cyanoaniline (35 mg, 0.3 mmol), gave 36 mg of a pale yellow solid. Yield: 57%; Melting point: 223 - 225 °C. 1 H NMR (600 MHz, d-CDCl3) δ 7.94 (d, J = 9.0 Hz, 2H), 7.58 - 7.63 (m, 4H), 7.48 - 7.44 (m, 2H), 7.05 (d, J = 8.4 Hz, 1H), 6.84 (dd, J = 7.8, 1.8 Hz, 1H), 6.76 (t, J = 7.8 Hz, 1H), 4.12 (s, 3H), 3.70 (s, 3H), 2.64 (s, 3H). HR-ESI-MS m / z 420.194 1 [M+H] + (Calculated value 420.193 1, C 25 H 22 N7).

[0178] Example 19: 2-[4-(2-Methoxyethoxy)anilino]-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 19, Method III)

[0179]

[0180] 2-Chloro-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 1, 51 mg, 0.15 mmol), 4-(2-methoxyethoxy)aniline (100 mg, 0.6 mmol), gave 58 mg of a light brown solid. Yield: 83%; Melting point: 160 - 162 °C. 1 H NMR (500 MHz, d-DMSO) δ (ppm) 9.16 (s, 1H), 7.85 (d, J = 9.0 Hz, 2H), 7.72 (d, J = 8.5 Hz, 1H), 7.43 - 7.37 (m, 2H), 7.29 (d, J = 1.0 Hz, 1H), 6.95 (d, J = 9.0 Hz, 1H), 6.91 - 6.86 (m, 3H), 6.70 - 6.86 (m, 1H), 4.06 (t, J = 5.0 Hz, 2H), 4.02 (s, 3H), 3.65 (t, J = 5.0 Hz, 2H), 3.58 (s, 3H), 3.32 (s, 3H), 2.60 (s, 3H). HR-ESI-MS m / z 469.235 3 [M+H] + (Calculated value 469.2347, C 27 H 29 N6O2).

[0181] Example 20: 2-(3-Aminomethanesulfonyl-4-methyl)-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 20, Method III)

[0182]

[0183] 2-Chloro-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 1, 51 mg, 0.15 mmol), 5-amino-2-methylbenzenesulfonamide (56 mg, 0.3 mmol), gave 43 mg of a white solid. Yield: 59%; Melting point: 215 - 217 °C. 11H NMR (600 MHz, d-DMSO) δ (ppm) 9.55 (s, 1H), 8.78 (s, 1H), 7.88 (d, J = 8.4 Hz, 1H), 7.73 (d, J = 8.4 Hz, 1H), 7.49 - 7.42 (m, 2H), 7.31 (s, 1H), 7.28 - 7.25 (m, 3H), 6.96 (d, J = 9.0 Hz, 1H), 6.89 (d, J = 9.0 Hz, 1H), 6.723 (t, J = 7.8 Hz, 1H), 4.03 (s, 3H), 3.61 (s, 3H), 2.61 (s, 3H), 2.53 (s, 3H). HR-ESI-MS m / z 488.186 6 [M+H] + (Calcd 488.186 3, C 25 H 26 N7O2S).

[0184] Example 21: 2-(4-Morpholinosulfonanilino)-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 21, Method III)

[0185]

[0186] 2-Chloro-4-(N,2,3-trimethyl-2H-indazol-6-ylamino)quinazoline (Compound 1, 51 mg, 0.15 mmol), 4-(Morpholinosulfonyl)aniline (73 mg, 0.3 mmol), gave 25 mg of a pale pink solid. Yield: 31%; Melting point: 259 - 261 °C. 1 1H NMR (600 MHz, d-DMSO) δ (ppm) 9.91 (s, 1H), 8.24 (d, J = 8.4 Hz, 2H), 7.75 (d, J = 8.4 Hz, 1H), 7.65 (d, J = 9.0 Hz, 2H), 7.54 (d, J = 8.4 Hz, 1H), 7.49 (t, J = 7.2 Hz, 1H), 7.36 (s, 1H), 7.00 (d, J = 9.0 Hz, 1H), 6.91 (d, J = 9.0 Hz, 1H), 6.79 (t, J = 7.2 Hz, 1H), 4.03 (s, 3H), 3.63 - 3.64 (m, 7H), 2.86 (m, 4H), 2.62 (s, 3H). HR-ESI-MS m / z 544.213 7 [M+H] + (Calcd 544.212 5, C 28 H 30 N7O3S).

[0187] Method 4: Palladium acetate (10% equiv) and X-Phos (20% equiv) were dissolved in a mixed solvent of 3 - 10 mL of 1,4-dioxane and water (5:1 v:v). Stir for 15 min at room temperature under N2 protection, then successively add N,2,3-trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1, 1 equiv), arylboronic acid (1.2 - 2 equiv) and cesium carbonate (1.4 - 4 equiv), and react for 30 min - 8 h under heating or microwave conditions. Pour the reaction mixture into an appropriate amount of ice water, precipitate, filter by suction, dry, and separate the crude product by Flash column chromatography to obtain the target product.

[0188] Example 22: N,2,3-Trimethyl-N-(2-(4-cyanophenyl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 22, Method 4)

[0189]

[0190] N,2,3-Trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1, 51 mg, 0.15 mmol), 4-cyanophenylboronic acid (30 mg, 0.2 mmol), palladium acetate (2 mg), X-Phos (9.5 mg), Cs2CO3 (130 mg, 0.4 mmol), 5 mL of 1,4-dioxane, 1 mL of water. React at 80 °C for 12 h. The crude product was separated by Flash neutral alumina column chromatography (AcOEt:PE = 0 - 50%) to obtain 37 mg of a yellow solid. Yield: 61%; Melting point: 218 - 220 °C. 1 H NMR (600 MHz, CDCl3) δ (ppm) 8.81 (s, 2H), 7.82 (d, J = 8.4 Hz, 2H), 7.60 (t, J = 8.4 Hz, 2H), 7.49 (s, 1H), 7.17 (d, J = 7.8 Hz, 1H), 6.98 (t, J = 7.2 Hz, 1H), 6.86 (dd, J = 8.4, 1.5 Hz, 1H), 4.12 (s, 3H), 3.84 (s, 3H), 2.65 (s, 3H). HR-ESI-MS m / z 405.183 4 [M + H] + (Calculated value 405.182 2, C 25 H 21 N6).

[0191] Example 23: N,2,3-Trimethyl-N-(2-(3-cyanophenyl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 23, Method 4)

[0192]

[0193] N,2,3-Trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1, 101 mg, 0.3 mmol), 3-cyanophenylboronic acid (55 mg, 0.36 mmol), palladium acetate (5.0 mg, 0.02 mmol), X-Phos (21 mg, 0.04 mmol), Cs2CO3 (138 mg, 0.42 mmol), 1,4-dioxane 3 mL, water 0.6 mL. Reacted by microwave heating at 120 °C for 1 h. The crude product was separated by Flash column chromatography (silica gel column, AcOEt∶(DCM∶PE = 1∶1) = 0 - 36%), and 101 mg of yellow solid was obtained. Yield: 83%; Melting point: 200 - 202 °C. 1 H NMR (600 MHz, CDCl3) δ 8.96 (s, 1H), 8.92 (brs, 1H), 7.92 (s, 1H), 7.77 (d, J = 7.2 Hz, 1H), 7.64 (t, J = 7.2 Hz, 1H), 7.60 - 7.59 (m, 2H), 7.48 (s, 1H), 7.18 (d, J = 8.4 Hz, 1H), 6.96 (t, J = 7.8 Hz, 1H), 6.87 (dd, J = 8.4, 1.8 Hz, 1H), 4.12 (s, 3H), 3.83 (s, 3H), 2.65 (s, 3H). HR-ESI-MS m / z 405.183 4 [M+H] + (Calcd 405.182 2, C 25 H 21 N6).

[0194] Example 24: N,2,3-Trimethyl-N-(2-(3-fluoro-4-cyanophenyl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 24, Method Four)

[0195]

[0196] N,2,3-Trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1, 67 mg, 0.2 mmol), 3-fluoro-4-cyanobenzeneboronic acid (40 mg, 0.24 mmol), palladium(II) acetate (4.5 mg, 0.02 mmol), X-Phos (19 mg, 0.04 mmol), Cs2CO3 (91 mg, 0.28 mmol), 1,4-dioxane 3 mL, water 0.6 mL. The reaction was heated to 120 °C by microwave for 1 h. The crude product was separated by flash column chromatography (silica gel column, AcOEt∶PE = 0 - 30%), and 65 mg of yellow solid was obtained. Yield: 77%; Melting point: 255 - 256 °C. 1 H NMR(600MHz,CDCl3)δ8.58(brs,1H),8.51(d,J=10.2Hz,1H),7.91(brs,1H),7.76(t,J=7.2Hz,1H),7.61~7.56(m,2H),7.47(s,1H),7.18(d,J=8.4Hz,1H),6.98(t,J=7.8Hz,1H),6.86(dd,J=9.0,1.8Hz,1H),4.12(s,3H),3.81(s,3H),2.65(s,3H).HR-ESI-MS m / z 423.174 0[M+H] + (Calculated value 423.172 8,C 25 H 20 FN6).

[0197] Example 25: N,2,3-Trimethyl-N-(2-(4-methylsulfonylphenyl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 25, Method Four)

[0198]

[0199] N,2,3-Trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1, 67 mg, 0.2 mmol), 4-methylsulfonylbenzeneboronic acid (48 mg, 0.24 mmol), palladium(II) acetate (4.5 mg, 0.02 mmol), X-Phos (20 mg, 0.04 mmol), Cs2CO3 (91 mg, 0.28 mmol), 1,4-dioxane 3 mL, water 0.6 mL. The reaction was heated to 120 °C by microwave for 1 h. The crude product was separated by flash column chromatography (silica gel column, AcOEt∶(DCM∶PE = 1∶1) = 0 - 50%), and 59 mg of yellowish brown solid was obtained. Yield: 64%; Melting point: 273 - 275 °C. 11H NMR (600 MHz, CDCl3) δ 8.86 (d, J = 7.2 Hz, 2H), 8.10 (d, J = 8.4 Hz, 2H), 7.59 (t, J = 7.8 Hz, 2H), 7.49 (s, 1H), 7.19 (d, J = 7.8 Hz, 1H), 6.98 (t, J = 7.8 Hz, 1H), 6.87 (dd, J = 9.0, 1.8 Hz, 1H), 4.12 (s, 3H), 3.84 (s, 3H), 3.12 (s, 3H), 2.65 (s, 3H). HR-ESI-MS m / z 458.165 6 [M+H] + (Calcd. for 458.1645, C 25 H 24 N5O2S).

[0200] Example 26: N,2,3-Trimethyl-N-(2-(3-furyl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 26, Method Four)

[0201]

[0202] N,2,3-Trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1, 101 mg, 0.3 mmol), pinacol 3-furylboronate (69 mg, 0.36 mmol), palladium(II) acetate (5.0 mg, 0.02 mmol), X-Phos (21 mg, 0.04 mmol), Cs2CO3 (137 mg, 0.42 mmol), 1,4-dioxane 3 mL, water 0.6 mL. The reaction mixture was heated to 120 °C by microwave irradiation for 30 min. The crude product was purified by flash column chromatography (silica gel column, AcOEt∶(DCM∶PE = 1∶1) = 0 - 40%) to give 67 mg of a grayish brown solid. Yield: 91%; mp: 181 - 183 °C. 1 1H NMR (600 MHz, CDCl3) δ 8.36 (s, 1H), 7.82 (s, 1H), 7.59–7.48 (m, 3H), 7.45 (s, 1H), 7.23 (s, 1H), 7.16 (d, J = 8.5 Hz, 1H), 6.89 (t, J = 7.6 Hz, 1H), 6.83 (dd, J = 8.8, 1.7 Hz, 1H), 4.10 (s, 3H), 3.75 (s, 3H), 2.63 (s, 3H). HR-ESI-MS m / z 370.167 1 [M+H] + (Calcd. for 370.166 2, C 22 H 20 N5O).

[0203] Example 27: N,2,3-Trimethyl-N-(2-(benzofuran-2-yl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 27, Method Four)

[0204]

[0205] N,2,3-Trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1, 67 mg, 0.2 mmol), benzofuran-2-boronic acid (39 mg, 0.24 mmol), palladium acetate (4.5 mg, 0.02 mmol), X-Phos (19 mg, 0.04 mmol), Cs2CO3 (92 mg, 0.28 mmol), 1,4-dioxane 3 mL, water 0.6 mL. Heated by microwave to 120 °C and reacted for 30 min. The crude product was separated by Flash column chromatography (silica gel column, AcOEt∶(DCM∶PE = 1∶1) = 0 - 30%), and 65 mg of light yellow solid was obtained. Yield: 77%; Melting point: 253 - 255 °C. 1 1H NMR (600 MHz, CDCl3) δ 8.06 (brs, 1H), 7.85 (brs, 1H), 7.73 - 7.70 (m, 2H), 7.59 - 7.55 (m, 2H), 7.49 (s, 1H), 7.40 (t, J = 7.8 Hz, 1H), 7.30 (t, J = 7.8 Hz, 1H), 7.18 (d, J = 8.4 Hz, 1H), 6.97 - 6.93 (m, 1H), 6.87 (dd, J = 8.4, 1.8 Hz, 1H), 4.11 (s, 3H), 3.83 (s 3H), 2.64 (s, 3H). HR-ESI-MS m / z 420.182 9 [M+H] + (Calcd 420.181 9, C 26 H 22 N5O).

[0206] Example 28: N,2,3-Trimethyl-N-(2-(3-furyl)quinazolin-4-yl)-2H-indazol-6-amine (Compound 28, Method Four)

[0207]

[0208] N,2,3-Trimethyl-N-(2-chloroquinazolin-4-yl)-2H-indazol-6-amine (Compound 1, 68 mg, 0.2 mmol), 2-fluoro-5-pyridineboronic acid (34 mg, 0.24 mmol), palladium acetate (4.5 mg, 2% mmol), X-Phos (19 mg, 4% mmol), Cs2CO3 (92 mg, 0.28 mmol), 1,4-dioxane 3 mL, water 0.6 mL. Reacted by microwave heating at 120 °C for 1 h. The crude product was separated by Flash column chromatography (silica gel column, AcOEt∶DCM = 0 - 40%), and 50 mg of yellow solid was obtained. Yield: 63%; Melting point: 253 - 254 °C. 1 H NMR (600 MHz, CDCl3) δ 9.45 (s, 1H), 9.03 (brs, 1H), 7.90 (brs, 1H), 7.60 - 7.56 (m, 2H), 7.48 (s, 1H), 7.17 (d, J = 7.2 Hz, 1H), 7.08 (d, J = 7.2 Hz, 1H), 6.95 (t, J = 7.8 Hz, 1H), 6.86 (dd, J = 9.0 & 1.8 Hz, 1H), 4.12 (s, 3H), 3.81 (s, 3H), 2.65 (s, 3H). HR-ESI-MS m / z 399.173 9 [M+H] + (Calculated value 399.172 8, C 23 H 20 FN6).

[0209] Example 29: In vitro tumor cell proliferation inhibition test of Compound of Formula 1

[0210] 1. Test method:

[0211] Human non-small cell lung cancer A549 cells, human liver cancer Huh-7 cells, human cervical cancer Hela cells, human glioma U251 cells, and human liver cancer HepG-2 cells were treated with different concentrations of the Compound of Formula I (prepared in Examples 1 - 28), incubated in an incubator at 37 °C and 5% CO2 for 72 h, and the inhibition rate of the compound on tumor cells was measured by the CCK-8 method. The positive control drug was colchicine, and the results are shown in Table 1.

[0212] 2. Test results:

[0213] Table 1 Proliferation inhibition activities of the compounds of the present invention against A549, Huh-7, Hela, U251, and HepG-2 tumor cells

[0214]

[0215] Note: ND: Not tested; A549: Human lung cancer cells; Huh-7: Human liver cancer cells; Hela: Human cervical cancer cells; U251: Human glioma cells; HepG-2: Human liver cancer cells.

[0216] Table 1

[0217] Example 30: In vitro tubulin aggregation inhibition experiment of Compound 2

[0218] 1. Test drugs: Compound 2 prepared in Example 2, Combretastatin (CA-4), and dimethyl sulfoxide (DMSO, control).

[0219] 2. Test method:

[0220] Mix different concentrations of porcine brain tubulin in PEM buffer (100 mM PIPES, 1 mM MgCl2, and 1 mM EGTA) containing 1 mM GTP and 5% glycerol. Using a SPECTRAMAX 190 spectrophotometer, monitor the tubulin polymerization process at 37 °C and 340 nm, and calculate the relative absorbance value.

[0221] 3. Test results:

[0222] See Figure 1 As shown, it can be seen from Figure 1 that Compound 2 can inhibit tubulin aggregation, similar to the mode of action of CA-4. The IC 50 value of Compound 2 for inhibiting tubulin aggregation is 0.78 ± 0.01 μM.

[0223] In the description of this specification, the description referring to terms such as "one embodiment", "some embodiments", "example", "specific example", or "some examples" means that the specific features, structures, materials, or characteristics described in connection with that embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, without contradiction, those skilled in the art can combine and combine the different embodiments or examples described in this specification and the features of different embodiments or examples.

[0224] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the protection scope of the present invention.

Claims

1. N,2,3-trimethyl-N-fused aryl-2H-indazol-6-amine compounds, characterized in that, Its structural formula is shown in Formula I: Wherein, X is N or CH; R 1 is H, halogen, cyano, carboxyl, C 1～6 alkyl, halo C 1～6 alkyl, C 3～12 cycloalkyl, halo C 3～12 cycloalkyl, hydroxyl, hydroxyl-substituted C 1～6 alkyl, hydroxyl-substituted C 3～12 cycloalkyl, C 1～6 alkoxy, C 3～12 cycloalkoxy, amino, C 1～6 alkylamino, C 3～12 cycloalkylamino, C 2～6 azacycloalkylamino, halogen-substituted C 2～6 azacycloalkylamino, hydroxyl-substituted C 1～6 alkylamino, hydroxyl-substituted C 3～12 cycloalkylamino, hydroxyl-substituted C 2～6 azacycloalkylamino, amino-substituted C 1～6 alkylamino, amino-substituted C 3～12 cycloalkylamino, amino-substituted C 2～6 azacycloalkylamino, hydroxyl-substituted C 1～6 dialkylamino, amino-substituted C 1～6 dialkylamino, heterocycloalkyl, aryl, arylamino, heteroaryl or heteroarylamino, wherein the aryl or heteroaryl ring optionally contains 0 to 2 N atoms, or simultaneously contains 0 to 1 nitrogen atom and oxygen atom, or simultaneously contains 0 to 1 nitrogen atom and sulfur atom; the aryl or heteroaryl is optionally substituted by 1 or 2 R 5 substituents; R 2 、R 3 、R 4 are each independently H, C 1～4 alkyl, C 1～4 alkoxy, halogen, cyano, hydroxy, C 1～4 alkylamino, C 1～4 dialkylamino, C 3～6 cycloalkyl or C 3～6 cycloalkylamino; R 5 independently H, C 1～4 alkyl, C 1～4 alkoxy, halogen, cyano, hydroxy, C 1～4 alkylamino, C 1～4 dialkylamino, N-hydroxycarbamoyl-substituted C 0～5 alkyl, N-hydroxycarbamoyl-substituted C 0～5 alkoxy, N-hydroxycarbamoyl-substituted C 0～5 alkylamino, hydrazinocarbonyl-substituted C 0～5 alkyl, hydrazinocarbonyl-substituted C 0～5 alkoxy, hydrazinocarbonyl-substituted C 0～5 alkylamino, C 3～6 cycloalkyl, C 3～6 cycloalkylamino, C 3～6 cyclic amino group containing 1 to 2 heteroatoms, C 3～6 cycloalkylamino group containing 1 to 2 heteroatoms, C 1～6 alkylsulfone, C 1～6 alkylaminomethanesulfonyl or C 1～6 heterocycloalkylaminomethanesulfonyl.

2. The N,2,3-trimethyl-N-fused aryl-2H-indazol-6-amine compound according to claim 1, characterized in that: X is N or CH; R 1 is H, halogen, cyano, carboxyl, C 1～6 alkyl, halo C 1～6 alkyl, C 3～12 cycloalkyl, halo C 3～12 cycloalkyl, hydroxy, hydroxy-substituted C 1～6 alkyl, hydroxy-substituted C 3～12 cycloalkyl, C 1～6 alkoxy, C 3～12 cycloalkoxy, amino, C 1～6 alkylamino, C 3～12 cycloalkylamino, C 2～6 azacycloalkylamino, halo-substituted C 2～6 azacycloalkylamino, hydroxy-substituted C 1～6 alkylamino, hydroxy-substituted C 3～12 cycloalkylamino, hydroxy-substituted C 2～6 azacycloalkylamino, amino-substituted C 1～6 alkylamino, amino-substituted C 3～12 cycloalkylamino, amino-substituted C 2～6 azacycloalkylamino, hydroxy-substituted C 1～6 dialkylamino, amino-substituted C 1～6 dialkylamino, heterocycloalkyl, aryl, arylamino, heteroaryl or heteroarylamino, wherein the aryl or heteroaryl ring optionally contains 0 to 2 N atoms, or simultaneously contains 0 to 1 nitrogen atom and oxygen atom, or simultaneously contains 0 to 1 nitrogen atom and sulfur atom; the aryl or heteroaryl is optionally substituted by 1 or 2 R 5 substituents; R 2 、R 3 、R 4 are each independently H, C 1～4 alkyl, C 1～4 alkoxy, halogen, cyano, hydroxy, C 1～4 alkylamino, C 1～4 dialkylamino, C 3～6 cycloalkyl or C 3～6 cycloalkylamino; R 5 independently H, C 1～4 alkyl, C 1～4 alkoxy, halogen, cyano, hydroxy, C 1～4 alkylamino, C 1～4 dialkylamino, N-hydroxycarbamoyl-substituted C 0～5 alkyl, N-hydroxycarbamoyl-substituted C 0～5 alkoxy, N-hydroxycarbamoyl-substituted C 0～5 alkylamino, hydrazinocarbonyl-substituted C 0～5 alkyl, hydrazinocarbonyl-substituted C 0～5 alkoxy, hydrazinocarbonyl-substituted C 0～5 alkylamino, C 3～6 cycloalkyl, C 3～6 cycloalkylamino, C 3～6 cyclic amino group containing 1 to 2 heteroatoms, C 3～6 cycloalkylamino group containing 1 to 2 heteroatoms, C 1～6 alkylsulfone, C 1～6 alkylaminomethanesulfonyl or C 1～6 heterocycloalkylaminomethanesulfonyl.

3. The N,2,3-trimethyl-N-fused aryl-2H-indazol-6-amine compound according to claim 1, characterized in that, Its structural formula is:

4. The N,2,3-trimethyl-N-fused aryl-2H-indazol-6-amine compound according to claim 1, wherein: The drug uses the compound or its pharmaceutically acceptable salt as the drug active component.

5. Preparation method of 5.N,2,3-trimethyl-N-fused aryl-2H-indazol-6-amine compounds, applied to the N,2,3-trimethyl-N-fused aryl-2H-indazol-6-amine compounds described in any one of claims 1 to 4, characterized in that, The preparation method is carried out according to the following reaction route: wherein R 6 is hydrogen or methyl, and optionally converting the resulting compound of formula I into a pharmaceutically acceptable salt thereof.

6. The preparation method according to claim 5, characterized in that, The preparation method comprises the following steps: in the presence of a base or an acid, reacting a compound in which R in general formula II 1 is substituted by a chlorine atom with a compound of general formula III (R 6 is H), 2,3-dimethyl-6-aminoindazole, in a solvent, and then introducing a methyl group onto the secondary amine N atom through a methylation reagent to obtain compounds 1 to 8 of general formula I; Or in the presence of a base or an acid, reacting a compound substituted with a chlorine atom of general formula II with a compound of general formula III (R 6 is methyl) N,2,3-trimethyl-6-aminoindazole in a solvent to obtain compounds 1 to 8 of general formula I, and subjecting compound 1 to dehalogenation, or substitution with a fatty amine / aromatic amine, or coupling with an aromatic ring compound to obtain compounds 9 to 28 of general formula I.

7. Use of the N,2,3-trimethyl-N-fused aryl-2H-indazole-6-amine compound according to any one of claims 1 to 3, characterized in that: The amine compound is used in combination with a pharmaceutically acceptable carrier.

8. The use according to claim 7, wherein: The use of the amine compound in anti-tumor drugs.

9. The use according to claim 8, characterized in that: The tumor is any one of blood cancer, ovarian cancer, prostate cancer, testicular cancer, melanoma, pancreatic cancer, lymphoma, breast cancer, gastric cancer, brain cancer, kidney cancer, lung cancer, liver cancer or colon cancer.

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