RIPK1 kinase target inhibitor as well as preparation method and application thereof

Abstract

The invention discloses an RIPK1 kinase target inhibitor as well as a preparation method and application thereof, and relates to the technical field of biological medicines. The preparation method comprises the following steps: respectively obtaining a quinazoline acid amide derivative (fragment A) and different benzo five-membered heterocyclic-2-amide derivatives (fragments B, C, D and E) through chemical synthesis, and carrying out coupling reaction on the quinazoline acid amide derivative and the different benzo five-membered heterocyclic-2-amide derivatives to obtain a target product as shown in a formula (I) or a formula (II). The derivatives taking the conjugates as the skeleton have a good RIPK1 inhibition effect, have a good programmed necrosis resistance effect, an acute ischemic stroke resistance effect and an acute colitis resistance effect, and are expected to have a good application prospect in the fields of neurodegenerative diseases, inflammations and autoimmune diseases.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a RIPK1 kinase target inhibitor and a preparation method and application thereof. Background Art

[0002] Programmed necrosis is a highly regulated process involving various signaling pathways, with core proteins including receptor-interacting serine / threonine-protein kinase 1 (RIPK1), RIPK3, mixed lineage kinase domain-like pseudokinase (MLKL), and cysteine ​​protease 8 (Caspase-8). Programmed necrosis mediates immune responses by activating RIPK1, RIPK3, and MLKL. RIPK1 contributes to the production of cytokines and DAMPs mediated by nuclear factor-κB (NF-κB) kinase subunit α (IKK-α) or TBK1 (TANK-binding kinase 1). MLKL maximizes the release of the above factors by forming plasma membrane pores. This process ultimately leads to morphological changes in cells, with cell swelling, plasma membrane rupture, and outflow of contents. RIPK1 is widely involved in regulating cell death and survival signaling pathways, especially playing an important role in inflammatory response and immune regulation.

[0003] During the onset of acute ischemic stroke (AIS), cerebral blood flow in the infarct area is significantly reduced. Sugar, oxygen supply and energy depletion can induce multiple signal cascades, causing programmed necrosis of nearby neurons, which is an important pathway of cell death. Its morphological characteristics are cell swelling, plasma membrane rupture and eventual release of cell contents, which are important pathological features of AIS. Degterev et al. first reported that programmed necrosis is involved in cerebral ischemia, abnormal expression of phosphorylated RIPK1, promoting inflammation and aggravating brain damage. When RIPK1 is inhibited, the degree of brain damage can be significantly reduced. The first selective programmed necrosis inhibitor Necrostatin-1 (Nec-1) was discovered. In addition, intraventricular administration of Nec-1 can significantly reduce infarct volume, indicating that Nec-1 has therapeutic potential for AIS. More and more studies have shown that the development of highly active and highly selective inhibitors targeting RIPK1 can be a very important therapeutic target for the treatment of acute ischemic stroke, providing a new approach for the treatment of this disease.

[0004] Studies have shown that RIPK1 is associated with a variety of neurological diseases, systemic inflammatory diseases, tumors, sepsis, etc., and phosphorylated RIPK1 will also be abnormally expressed. The application of RIPK1 inhibitors is expected to provide a new approach for the treatment of neurological diseases, systemic inflammatory diseases, tumors, sepsis, etc. Summary of the invention

[0005] Based on this, the purpose of the present invention is to overcome the deficiencies of the above-mentioned prior art and provide a RIPK1 kinase target inhibitor and a preparation method and application thereof.

[0006] To achieve the above object, the technical solution adopted by the present invention is: a RIPK1 kinase target inhibitor, the general structural formula of the RIPK1 kinase target inhibitor is shown in formula (I) or formula (II):

[0007]

[0008] Wherein, X, Y, and Z are all nitrogen atoms; or, Z is a carbon atom, X and Y are one of a nitrogen atom and a sulfur atom, and the atoms of X and Y are different; or, Z is a carbon atom, and X and Y are both nitrogen atoms; or, Y and Z are carbon atoms, and X is an oxygen, nitrogen, or sulfur atom;

[0009] Among them, R 1 is selected from one of hydrogen, methyl, ethyl, propyl, isopropyl, phenyl, and substituted phenyl; R 2 and R 3 Each is independently selected from an alkyl group having less than 6 carbon atoms or a halogen-substituted product thereof, a cycloalkyl group having less than 6 carbon atoms or a halogen-substituted product thereof, a phenyl group or a halogen-substituted product thereof, a benzyl group, a substituted benzyl group, a methylene group substituted with an aromatic heterocycle or a halogen-substituted product thereof;

[0010] Wherein, G in formula (II) is one of CH2, NH, and O.

[0011] Preferably, X, Y, and Z are nitrogen atoms at the same time to form a triazole ring, or Z is a carbon atom, X and Y are one of a nitrogen atom and a sulfur atom, and the atoms of X and Y are different to form a thiazole ring; or Z is a carbon atom, and X and Y are both nitrogen atoms to form an imidazole ring; or Y and Z are carbon atoms, and X is an oxygen, nitrogen or sulfur atom to form a furan ring, a pyrrole ring or a thiophene ring.

[0012] Preferably, in the general structural formula such as formula (I) or formula (II), X, Y, and Z are nitrogen atoms at the same time to form a triazole ring; or Z is a carbon atom, and X and Y are one of a nitrogen atom and a sulfur atom, and the atoms of X and Y are selected differently to form a thiazole ring.

[0013] Preferably, the structural formula of the RIPK1 kinase target inhibitor is selected from at least one of the following:

[0014]

[0015]

[0016]

[0017] Preferably, the RIPK1 kinase target inhibitor is selected from at least one of the following: (R)-6-(2-isobutyramido-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide, (S)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide, (R)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline- 4-carboxamide, (S)-6-(2-isobutylamido-[1,2,4]triazolyl[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide, (S)-2-methyl-N-(1-phenylethyl)-6-(2-neoamido-[1,2,4]triazolo[1,5-a]pyridin-7-yl)quinazoline-4-carboxamide, (S)-2-methyl-6-(2-pentanamido-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-N-(1-phenylethyl)quinazoline-4-carboxamide, (S)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)- quinazoline-4-carboxamide, (S)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-(pyridin-2-yl)ethyl)quinazoline-4-carboxamide, (S)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-(pyridin-3-yl)ethyl)quinazoline-4-carboxamide, (S)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-(pyridin-4-yl)ethyl)quinazoline-4-carboxamide, 1,5-a]pyridin-7-yl)-N-(1-(3-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide, (S)-6-(2-(cyclopentylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide, (S)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-N-(1-(2-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide, (S)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-N-(1-(2-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide

[0018] quinazoline-4-carboxamide, (S)-6-(2-(cyclohexylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(4-(trifluoromethyl)benzyl)quinazoline-4-carboxamide, (S)-6-(2-(cyclohexylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide, (S)-6-(2-(cyclohexylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide, (S)-6-(2-butylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(4-(trifluoromethyl)benzyl)quinazoline-4-carboxamide, quinazoline-4-carboxamide, 6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide, (S)-6-(2-(cyclobutanecarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(4-(trifluoromethoxy)benzyl)quinazoline-4-carboxamide, (S)-6-(2-(cyclobutanecarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide, 6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-( 3-(trifluoromethyl)benzyl)quinazoline-4-carboxamide, 6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(3-(trifluoromethoxy)benzyl)quinazoline-4-carboxamide, (S)-2-methyl-6-(2-(3-oxocyclobutane-1-carboxamido)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-N-(1-phenylethyl)quinazoline-4-carboxamide, (S)-N-(1-(4-fluorophenyl)ethyl)-2-methyl-6-(2-(3-oxocyclobutane-1-carboxamido)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)quinazoline Quinazoline-4-carboxamide, (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide, (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide, (S)-N-(1-(4-fluorophenyl)ethyl)-6-(2-isobutyramidobenzo[d]thiazol-6-yl)-2-methylquinazoline-4-carboxamide, (S)-N-(1-(4-fluorophenyl)ethyl)-2-methyl-6-(2-pentanamidobenzo[d]thiazol-6-yl)quinazoline-4-carboxamide,(S)-N-(1-(4-fluorophenyl)ethyl)-2-methyl-6-(2-(3-methylbutyramido)benzo[d]thiazol-6-yl)quinazoline-4-carboxamide, (S)-N-(1-(4-fluorophenyl)ethyl)-2-methyl-6-(2-pivalamidobenzo[d]thiazol-6-yl)quinazoline-4-carboxamide, (S)-6-(2-(cyclobutanecarboxamido)benzo[d]thiazol-6-yl)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide, (S)-6-(2-(cyclopentanecarboxamido)benzo[d]thiazol-6-yl)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide amine, (S)-6-(2-(cyclohexanecarboxamido)benzo[d]thiazol-6-yl)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide, (S)-6-(2-benzylaminobenzo[d]thiazol-6-yl)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide, (S)-6-(2-butyrylaminobenzo[d]thiazol-6-yl)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide, (S)-N-(1-(4-fluorophenyl)ethyl)-2-methyl-6-(2-propionamidobenzo[d]thiazol-6-yl)quinazoline-4-carboxamide, (S)-6-(2- acetylaminobenzo[d]thiazol-6-yl)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide, N-benzyl-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methylquinazoline-4-carboxamide, (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(1-phenylpropyl)quinazoline-4-carboxamide, 6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-phenylquinazoline-4-carboxamide, 6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-phenylethylquinazoline-4-carboxamide Quinazoline-4-carboxamide, (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(tetrahydro-2H-pyran-4-yl)quinazoline-4-carboxamide, (S)-N-(1-cyclohexylethyl)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methylquinazoline-4-carboxamide, (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(1-(p-tolyl)ethyl)quinazoline-4-carboxamide, (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(1-(pyridin-4-yl)ethyl)quinazoline-4-carboxamide,(S)-6-(2-(Cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-N-(1-(4-methoxyphenyl)ethyl)-2-methylquinazoline-4-carboxamide, (S)-6-(2-(Cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(2-methyl-1-phenylpropyl)quinazoline-4-carboxamide, (R)-6-(2-(Cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide, (S)-6-(2-(Cyclopropanecarboxamido)benzo[d]thiazol-6-yl) -2-methyl-N-(1-(4-nitrophenyl)ethyl)quinazoline-4-carboxamide, (S)-6-((2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)methyl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide, (S)-6-((2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)amino)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide, (S)-6-((2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)oxy)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide.

[0019] Preferably, the RIPK1 kinase target inhibitor is selected from (S)-6-(2-(cyclopropylcarboxamido)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide, (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide, (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide,

[0020] (S)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide, (S)-N-(1-(4-fluorophenyl)ethyl)-2-methyl-6-(2-propionamidobenzo[d]thiazol-6-yl)quinazoline-4-carboxamide, (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(1-phenylpropyl)quinazoline Quinazoline-4-carboxamide, (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(1-(p-tolyl)ethyl)quinazoline-4-carboxamide, (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-N-(1-(4-methoxyphenyl)ethyl)-2-methylquinazoline-4-carboxamide.

[0021] The inventors found in the actual experimental process that the above-mentioned RIPK1 kinase target inhibitors have better comprehensive effects, and the superior compounds come from the comprehensive results of enzymology and cell experiments.

[0022] In addition, the present invention provides a method for preparing the RIPK1 kinase target inhibitor, comprising the following steps:

[0023] (1) Synthesis of 6-bromoquinazoline acid amide derivatives: 5-bromoisatin is used as a raw material, subjected to alkaline hydrolysis, and then diluted hydrochloric acid is added to adjust the pH to neutral to obtain a sodium salt of the indigo carboxylate hydrolyzate; the sodium salt of the indigo carboxylate hydrolyzate is subjected to a three-component reaction with acetaldehyde and ammonium acetate to obtain a sodium salt of quinazoline carboxylate; in the presence of a condensing agent, the sodium salt of quinazoline carboxylate is condensed with an amine compound to generate a 6-bromoquinazoline amide derivative, which is fragment A;

[0024] (2) At least one of the following (2a) to (2d):

[0025] (2a) Preparation of fragment B: Using a 7- or 6-brominated 2-amino-benzo five-membered heterocyclic compound as a raw material, adding an acyl chloride or a carboxylic acid, and in the presence of an acid binding agent or a condensing agent, preparing a 7- or 6-brominated benzo five-membered heterocyclic-2-amino acylated derivative as fragment B;

[0026] (2b) Preparation of fragment C: Using a 2-amino-benzo five-membered heterocyclic compound brominated at the 6-position as a raw material, adding a carboxylic acid, and in the presence of a condensing agent, preparing a benzo five-membered heterocyclic 2-aminoacylated derivative substituted with a bromomethyl group at the 6-position, namely, fragment C;

[0027] (2c) preparing fragment D, using 2-amino-benzo five-membered heterocycle substituted with nitro at the 6-position as a raw material, adding carboxylic acid, in the presence of a condensing agent, preparing a benzo five-membered heterocycle substituted with nitro at the 6-position-2-aminoacylated derivative, adding a nitro reducing agent, preparing the corresponding benzo five-membered heterocycle substituted with amino at the 6-position-2-aminoacylated derivative, namely fragment D;

[0028] (2d) preparing fragment E, using 2-amino-benzo five-membered heterocycle substituted with 6-hydroxyl group as raw material, adding phenolic hydroxyl protecting agent to protect the hydroxyl group, adding carboxylic acid, preparing 6-silyl ether protected benzo five-membered heterocycle-2-amino acylated derivative in the presence of condensing agent, adding deprotecting agent to prepare the corresponding 6-phenolic hydroxyl substituted benzo five-membered heterocycle-2-amino acylated derivative, namely fragment E;

[0029] (3) At least one of the following (3a) to (3c):

[0030] (3a) reacting the fragment B prepared in step (2) with pinacol diboron in the presence of an organic base A and a palladium catalyst to prepare an aryl borate of the fragment B, and then reacting the fragment A prepared in step (1) in the presence of an organic base B, a palladium catalyst and a phosphine ligand to obtain a RIPK1 kinase target inhibitor represented by formula (I); the organic base A is potassium acetate, and the organic base B is at least one of Cs2CO3 and K3PO4;

[0031] (3b) reacting the fragment A prepared in step (1) with pinacol diboron in the presence of an organic base A and a palladium catalyst to prepare the corresponding aryl borate, and then reacting it with the fragment C prepared in step (2) in the presence of an organic base B, a palladium catalyst and a phosphine ligand to obtain a RIPK1 kinase target inhibitor represented by formula (II);

[0032] (3c) reacting the fragment D or fragment E prepared in step (2) with the fragment A prepared in step (1) in the presence of an organic base B, a palladium catalyst, and a phosphine ligand to obtain a RIPK1 kinase target inhibitor represented by formula (II).

[0033] Preferably, in step (2), the synthesis route of fragment A is as follows (i):

[0034]

[0035] Preferably, in step (2), the synthesis route of fragment B is as follows (ii):

[0036]

[0037]

[0038] Preferably, in step (2), the synthesis route of fragment C is as follows (iv):

[0039]

[0040] Preferably, in step (2), the synthesis route of fragment D is as follows (v):

[0041]

[0042] Preferably, in step (2), the synthesis route of fragment E is as follows (vi):

[0043]

[0044] Preferably, in step (1), the base is at least one of sodium hydroxide and potassium hydroxide;

[0045] And / or, the condensing agent is one of mixture A and mixture B, the mixture A is a mixture of N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate and N-methylimidazole, and the molar ratio of the mixture of N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate and N-methylimidazole is 1:2-3; the mixture B is a mixture of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 1-hydroxybenzotriazole, and triethylamine, and the molar ratio of them is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride:1-hydroxybenzotriazole:triethylamine=1:1:1;

[0046] and / or, the amine compound is (R)-1-phenylethylamine, (S)-1-phenylethylamine, (S)-1-phenylpropylamine, (S)-2-methyl-1-phenylpropylamine, (S)-1-cyclohexylethylamine, (S)-1-(4-methylphenyl)ethylamine, (S)-1-(4-methoxyphenyl)ethylamine, (S)-1-(4-nitrophenyl)ethylamine, (S)-1-(pyridin-2-yl)ethylamine, (S)- At least one of 1-(pyridin-3-yl)ethylamine, (S)-1-(pyridin-4-yl)ethylamine, (S)-1-(2-fluorophenyl)ethylamine, (S)-1-(3-fluorophenyl)ethylamine, (S)-1-(4-fluorophenyl)ethylamine, aniline, benzylamine, phenethylamine, tetrahydro-2H-pyran-4-amine, 4-trifluoromethylbenzylamine, 3-trifluoromethylbenzylamine, 4-trifluoromethoxybenzylamine, and 3-trifluoromethoxybenzylamine;

[0047] And / or, the molar ratio of 5-bromoinsigmatine and base is 5-bromoinsigmatine:base=1:1-3;

[0048] And / or, the molar ratio of the sodium salt of isatin hydrolysate, acetaldehyde and ammonium acetate is: sodium salt of isatin hydrolysate: acetaldehyde: ammonium acetate = 1:1-3:2-5;

[0049] And / or, the molar ratio of the quinazoline carboxylic acid sodium salt, the amine compound, and the condensing agent is 1:1:2-3;.

[0050] Preferably, in steps (2a) to (2d):

[0051] The acyl chloride is at least one of n-butyryl chloride, isobutyryl chloride, n-valeryl chloride, pivaloyl chloride, cyclopropylcarbonyl chloride, cyclobutylcarbonyl chloride, cyclopentylcarbonyl chloride, cyclohexylcarbonyl chloride and 3-oxocyclobutylcarbonyl chloride;

[0052] and / or, the carboxylic acid is at least one of acetic acid, propionic acid, n-butyric acid, isobutyric acid, n-valeric acid, isovaleric acid, pivalic acid, cyclopropanecarboxylic acid, cyclobutanecarboxylic acid, cyclopentanecarboxylic acid, cyclohexanecarboxylic acid, and benzoic acid;

[0053] And / or, the acid binding agent is at least one of triethylamine and N,N-diisopropylethylamine;

[0054] And / or, the condensing agent is one of mixture A and mixture B, the mixture A is a mixture of N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate and N-methylimidazole, and the molar ratio of the mixture of N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate and N-methylimidazole is 1:2-3; the mixture B is a mixture of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 1-hydroxybenzotriazole, and triethylamine, and the molar ratio of them is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride:1-hydroxybenzotriazole:triethylamine=1:1:1;

[0055] And / or, the nitro reducing agent is stannous chloride;

[0056] And / or, the phenolic hydroxyl protecting agent is a mixture of tert-butyldimethylchlorosilane and imidazole, and the molar ratio of tert-butyldimethylchlorosilane to imidazole is 1:1-2;

[0057] And / or, the deprotection agent is tetrabutylammonium fluoride.

[0058] Preferably, in step (2a), the molar ratio of 2-amino-benzo five-membered heterocyclic compound, acyl chloride and acid binding agent is 2-amino-benzo five-membered heterocyclic compound: acyl chloride: acid binding agent = 1:1-2:1-3;

[0059] And / or, in the steps (2a), (2b), (2c) and (2d), the molar ratio of the 2-amino-benzo five-membered heterocyclic compound, the carboxylic acid and the condensing agent is 2-amino-benzo five-membered heterocyclic compound: carboxylic acid: condensing agent = 1:1:1.1-1.5;

[0060] And / or, in the step (2c), the molar ratio of the benzo five-membered heterocyclic 2-aminoacylated derivative substituted with nitro at the 6-position and the nitro reducing agent is benzo five-membered heterocyclic 2-aminoacylated derivative substituted with nitro at the 6-position: nitro reducing agent = 1:3-6;

[0061] And / or, in the step (2d), the molar ratio of the 2-amino-benzo five-membered heterocyclic ring substituted with the 6-hydroxyl group to the phenolic hydroxyl protecting agent is 1:1-3;

[0062] And / or, in the step (2d), the molar ratio of the 6-silyl ether protected benzo five-membered heterocyclic 2-aminoacylated derivative and the deprotection agent is 6-silyl ether protected benzo five-membered heterocyclic 2-aminoacylated derivative: deprotection agent = 1:2-5.

[0063] In the steps (3a), (3b) and (3c), the palladium catalyst is at least one of Pd(dppf)Cl2 and Pd(dppf)Cl2·CH2Cl2, and the phosphine ligand is PCy3;

[0064] And / or, in the step (3a), the molar ratio of the fragment B prepared in the step (2), the biphenyl pinacol ester, the palladium catalyst, and the organic base A is fragment B: biphenyl pinacol ester: palladium catalyst: organic base A = 1: 2-3: 0.01-0.3: 2-5;

[0065] And / or, in the step (3a), the molar ratio of the aryl borate of fragment B, fragment A, palladium catalyst, phosphine ligand, and organic base B is aryl borate of fragment B: fragment A: palladium catalyst: phosphine ligand: organic base B = 1: 1: 0.01-0.2: 0.01-0.2: 2-5;

[0066] And / or, in the step (3b), the molar ratio of the fragment A prepared in the step (1), the bipyraclostrobin, the palladium catalyst, and the organic base A is fragment A: bipyraclostrobin: palladium catalyst: organic base A = 1: 2-3: 0.01-0.3: 2-5;

[0067] And / or, in the step (3b), the molar ratio of the aryl borate of fragment A, fragment C, palladium catalyst, phosphine ligand, and organic base B is aryl borate of fragment A: fragment C: palladium catalyst: phosphine ligand: organic base B = 1: 1: 0.01-0.2: 0.01-0.2: 2-5;

[0068] And / or, in the step (3c), the molar ratio of fragment A prepared in step (1), fragment D or fragment E prepared in step (2), palladium catalyst, phosphine ligand, and organic base B is fragment A: fragment D / fragment E: palladium catalyst: phosphine ligand: organic base B = 1: 1: 0.01-0.2: 0.01-0.2: 2-5.

[0069] In the step (3), the fragment B prepared in the step (2) is reacted with biboronic acid pinacol ester in the presence of an organic base A and a palladium catalyst to prepare an aryl borate ester of the fragment B, and then reacted with the fragment A prepared in the step (1) in the presence of an organic base B, a palladium catalyst and a phosphine ligand to obtain the RIPK1 kinase target inhibitor represented by formula (I). Specifically, the synthetic routes are as follows: formula (vii) and formula (viii);

[0070]

[0071] Preferably, the fragment A prepared in step (1) is coupled with the fragments C, D, and E prepared in step (2) to obtain the RIPK1 kinase target inhibitor represented by formula (II). Specifically, the synthetic route is as follows:

[0072]

[0073] The present invention provides a pharmaceutical composition, wherein the pharmaceutical composition contains the RIPK1 kinase target inhibitor.

[0074] Preferably, the pharmaceutical composition is prepared into any one dosage form of injection, solid oral preparation, and sustained-release preparation.

[0075] Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0076] The present invention also provides the use of the RIPK1 kinase target inhibitor or the pharmaceutical composition in the preparation of drugs for treating neurodegenerative diseases, inflammation, autoimmune diseases, and sepsis.

[0077] Preferably, the inflammatory drug comprises an anti-acute colitis drug.

[0078] The present invention provides a medicine, which contains the RIPK1 kinase target inhibitor or the pharmaceutical composition.

[0079] Compared with the prior art, the beneficial effects of the present invention are as follows: the present invention obtains quinazole acid amide derivatives (fragment A) and different benzo five-membered heterocyclic-2-amide derivatives (fragments B, C, D and E) respectively by chemical synthesis, and couples the two to obtain the target product of formula (I) or formula (II). These conjugates are skeleton derivatives with good RIPK1 inhibition, good anti-programmed necrosis effect, anti-acute ischemic stroke effect and acute colitis effect, and are expected to have good application prospects in the fields of neurodegenerative diseases, inflammation and autoimmune diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0080] Figure 1 The effect of compound 1-23 on HT-29 cell viability, Note: Compared with the blank group, *p<0.05, **p<0.01, ***p<0.001, data are expressed as Mean±SEM;

[0081] Figure 2 The graph shows the protective effect of compound 2 on TSZ-induced programmed necrosis in HT-29, U937, L929, and HT-22 (AD) cells; Note: data are expressed as Mean±SEM.

[0082] Figure 3 The protective effect of compound 2 on TSZ-induced programmed necrosis in HT-29 (A), U937 (B), L929 (C), and HT-22 (D) cells is dose-dependent; Note: *p<0.05, **p<0.01, ***p<0.001, data are expressed as Mean±SEM;

[0083] Figure 4 The microscopic images show that compound 2 protects four types of cells from programmed necrosis;

[0084] Figure 5 Fluorescence staining of compound 2 protecting L929 cells from programmed necrosis;

[0085] Figure 6 This is a fluorescent staining image showing that compound 2 protects HT-29 cells from programmed necrosis;

[0086] Figure 7 Zea Longa (A) and MNSS (B) scores of compound 2 in rat MCAO model; Note: compared with the model group, *p<0.05, **p<0.01, ***p<0.001; n=9-12, data are expressed as Mean±SEM;

[0087] Figure 8 The statistical graphs of brain TTC staining (A) and percentage of cerebral infarction volume of animals in each group; Note: Compared with the model group, **p<0.01, ***p<0.001; n=9-12, data are expressed as Mean±SEM;

[0088] Fig. 9 This is the change diagram of serum SOD, MDA, TNF-α and IL-1β levels in rats 24h after MCAO reperfusion;

[0089] Fig.10 It is the oral and intravenous drug-time curve of compound 2;

[0090] Fig.11 For compound 2 1 H NMR spectrum. DETAILED DESCRIPTION

[0091] In order to better illustrate the purpose, technical scheme and advantages of the present invention, the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments. Its purpose is to understand the content of the present invention in detail, rather than to limit the present invention. All other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention. The experimental reagents and instruments designed for the implementation and comparative examples of the present invention are all commonly used ordinary reagents and instruments, unless otherwise specified, and can be obtained from commercial channels. In the implementation and comparative examples, the experimental methods used are conventional methods unless otherwise specified; and unless otherwise specified, the raw materials used in parallel experiments are the same batch of raw materials, and all the raw materials used in the present invention are commercially available and can be purchased from any one of Aladdin, McLean or Anage.

[0092] The embodiments of the present invention provide 51 methods for preparing RIPK1 kinase target inhibitors as shown in Examples 1 to 51. The structural formulas of the 51 RIPK1 kinase target inhibitors are as follows: Formula 1 to Formula 51. The following examples are only part of the embodiments of the present invention and should not be used to limit the scope of protection of the present invention. The target product can also be obtained by using other conditions in the preparation method of the pyridine amide compound described above in the present invention. However, due to space limitations, this article will not list them one by one.

[0093]

[0094]

[0095]

[0096] Example 1: Synthesis of (R)-6-(2-isobutyramido-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide

[0097] (1) Synthesis of sodium 6-bromo-2-methylquinazoline-4-carboxylate:

[0098] 5-Bromoindigo carmine (45.2 g, 200 mmol) was placed in a beaker and 2000 mL of water was added to stir the suspension, and then NaOH (8.8 g, 220 mmol) was added and stirred for 3 h to obtain the sodium salt of the indigo carmine hydrolyzate, and 2N HCl solution was slowly added dropwise to adjust the pH to 7. The aqueous solution was spin-dried and vacuum dried to obtain 52.0 g of sodium 2-(2-amino-5-bromophenyl)-2-oxoacetate, with a yield of 97%.

[0099] Sodium 6-bromo-2-methylquinazoline-4-carboxylate (51.9 g, 195 mmol), ammonium acetate (30.1 g, 390 mmol), ethanol (500 mL), and acetaldehyde-tetrahydrofuran solution (9.0 g, 205 mmol) were added to a 1000 mL round-bottom flask in sequence, and heated to 90°C under balloon protection to react. After 24 h, half the volume of the solvent was dried by spin-drying, and the mixture was allowed to stand at -18°C for 1 h to precipitate. The filter cake was washed with ethanol and ethyl acetate in sequence, and dried to obtain 22.7 g of sodium 6-bromo-2-methylquinazoline-4-carboxylate as a brown powder with a yield of 40%. 1 HNMR (400MHz, DMSO-d6) δ8.44(s,1H),7.98(dd,J=8.9,2.3Hz,1H),7.78(d,J=8.9Hz,1H),2.69(s,3H).

[0100] (2) Synthesis of (R)-6-bromo-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide:

[0101] Sodium 6-bromo-2-methylquinazoline-4-carboxylate (578 mg, 2.0 mmol), small molecule aromatic amine (2.2 mmol), dichloromethane (20 mL) and N-methylimidazole (411 mg, 5.0 mmol) were added to a 100 mL round-bottom flask in sequence, mixed thoroughly, and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (729.5 mg, 2.6 mmol) was slowly added under an ice-water bath. After the addition was complete, the ice bath was removed, the temperature was raised to room temperature, and the reaction was continued for another 2 h. The reaction was terminated after TLC monitoring of the reaction was complete. The reaction solution was extracted with ethyl acetate, the organic layer was dried, spin-dried, and purified by column chromatography to obtain compound (R)-6-bromo-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide as an off-white powder with a yield of 56%. 1 H NMR(400MHz,Chloroform-d)δ9.73(d,J=2.2Hz,1H),8.53(d,J=8.2Hz,1H),7.94(dd,J=9.0,2.2Hz,1H),7.83(d,J=9.0Hz,1 H),7.46-7.43(m,2H),7.39(t,J=7.6Hz,2H),7.35-7.26(m,1H),5.33(p,J=7.1Hz,1H),2.89(s,3H),1.68(d,J=6.9Hz,3H).

[0102] (3) Synthesis of N-(7-bromo-[1,2,4]triazolo[1,5-a]pyridin-2-yl)isobutyramide:

[0103] 7-Bromo-[1,2,4]triazolo[1,5-a]pyridine-2-amine (2.13 g, 10.0 mmol) was placed in a 100 mL round-bottom flask, and dichloromethane (50 mL) and triethylamine (1.11 g, 11.0 mmol) were added in sequence. The mixture was stirred under an ice-water bath, and isobutyryl chloride (10.1 mmol) was slowly added dropwise. After 2 h, the reaction was monitored by TLC. When the raw material point disappeared, the reaction was terminated. The dichloromethane was dried by spin drying, and ethyl acetate was added for extraction. The mixture was dried again by spin drying, and purified by column chromatography. The column chromatography product was added to a round-bottom flask, and 7M ammonia-methanol solution (50.0 mmol) was added under an ice-water bath, and stirred for 0.5 h. The reaction was monitored by TLC until it was converted into a monoacylated product. The solution was spin dried, ethyl acetate / petroleum ether was added to make a pulp, filtered, the filter cake was washed with water, and the filter cake was dried to obtain the compound N-(7-bromo-[1,2,4]triazolo[1,5-a]pyridin-2-yl)isobutyramide as a white powder with a yield of 60%. 1 H NMR (400MHz, DMSO-d6) δ10.78(s,1H),8.81(d,J=7.6Hz,1H),8.03(d,J=1.5Hz,1H),7.29(dd,J=7.1,2.1Hz,1H),2.52(s,1H),1.10(d,J=6.8Hz,6H).

[0104] (4) Synthesis of N-(7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)isobutyramide:

[0105] N-(7-Bromo-[1,2,4]triazolo[1,5-a]pyridin-2-yl)isobutyramide (707.8 mg, 2.50 mmol), bipyraclostrobin (952.3 mg, 3.75 mmol), and KOAc (736.1 mg, 7.50 mmol) were placed in a 100 mL completely dry round-bottom flask, and Pd(dppf)Cl2 (91.5 mg, 0.13 mmol) and anhydrous dioxane (30 mL) were added in sequence; the gas in the flask and spherical condenser was fully replaced with argon using a three-way valve and a balloon, and refluxed at 90°C for 12 h. After the reaction was completed as monitored by TLC, silica gel was added, the solvent was evaporated, and column chromatography was directly performed for purification to obtain N-(7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)isobutyramide as a light brown powder with a yield of 68%.

[0106] (5) (R)-6-(2-isobutylamido-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline

[0107] -Synthesis of 4-formamide:

[0108] (R)-6-Bromo-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide (370.3 mg, 1.0 mmol), N-(7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)isobutyramide (330.2 mg, 1.0 mmol), Pd(dppf)Cl2·CH2Cl2 (81.7 mg, 0.1 mmol), PCy3 (14.0 mg, 0.05 mmol), Cs2CO3 (814.6 mg, 2.50 mmol) and anhydrous dioxane (30 mL) were added into a 100 mL round-bottom flask in sequence. The gas in the flask and spherical condenser was fully replaced with argon using a three-way valve and a balloon, and then refluxed at 110°C for 3 h. After the reaction was completed as monitored by TLC, the solution was directly sanded in the original flask and purified by column chromatography to obtain (R)-6-(2-isobutyramido-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide.

[0109] The powder prepared in this example is a light yellow powder, and the structural formula is shown in Formula 1 above.

[0110] Yield: 75%, melting point: >220°C, purity: 97.9%.

[0111] Test method: The yield was calculated by weighing the mass of the purified target compound in ChemDraw software. The melting point of the target compound was tested in parallel three times on an automatic melting point instrument (OptiMelt MPA100, Stanford Research Systems, USA). The purity of the target compound was tested using UPLC. Chromatographic conditions: The compound purity was determined in an ultra-high performance liquid chromatograph equipped with an Agilent C8 column (2.1mm×50mm, 1.8μm), using a diode array (DAD) detector. The chromatographic conditions were initial MeOH / H2O=70 / 30, gradient elution, and the MeOH ratio gradually increased to 100% within 10min; the aqueous phase contained 10mM NH4OAc, and the pH was adjusted to 9 with ammonia water; the flow rate was 1.0mL / min; the retention time was 25min; the chromatographic temperature was set to 30℃; and the injection volume was 10μL each time.

[0112] The subsequent testing methods are the same and will not be described in detail here.

[0113] 1H NMR (400MHz, DMSO-d6) δ10.80(s,1H),9.47(d,J=8.2Hz,1H),8.96(d,J=7.1Hz,1H),8.71(d ,J=2.1Hz,1H),8.50(dd,J=8.9,2.2Hz,1H),8.11(d,J=8.9Hz,1H),8.01(d,J=1.9Hz,1H),7. 50(d,J=7.2Hz,2H),7.39(dt,J=7.1,5.1Hz,3H),7.30(t,J=7.3Hz,1H),5.30(p,J=7.1Hz,1 H),2.87-2.49(m,4H),1.56(d,J=7.0Hz,3H),1.13(d,J=6.8Hz,6H).HRMS(m / z):calculated for C 28 H 28 O2N7[M+H] + 494.22990, found 494.23013.

[0114] Example 2: Synthesis of (S)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide

[0115] The raw materials are cyclopropylcarbonyl chloride and (S)-1-phenylethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0116] The product prepared in this embodiment is a white powder, and its structural formula is shown in Formula 2 above.

[0117] Physical, chemical and spectral data: Yield: 87%, melting point: >278°C, purity: 96.9%. 1 H NMR (400MHz, DMSO-d6) δ11.12(s,1H),9.45(d,J=8.2Hz,1H),8.95(d,J=7.0Hz,1H),8.71 (d,J=2.1Hz,1H),8.49(dd,J=8.9,2.2Hz,1H),8.10(d,J=8.9Hz,1H),8.01(s,1H),7.50( d,J=7.4Hz,2H),7.45-7.34(m,3H),7.30(t,J=7.3Hz,1H),5.31(p,J=7.2Hz,1H),2.87(s ,3H),2.10(s,1H),1.56(d,J=7.0Hz,3H),0.86(d,J=5.4Hz,4H).HRMS(m / z):calculated for C28 H 26 O2N7[M+H] + 492.21425, found 492.21448.

[0118] Taking the compound 2 prepared in this example as an example, 1 H NMR spectrum is used for illustration.

[0119] Compound 2 1 H spectrum Fig.11 As shown, the d peaks at δ=11.12 and 8.95 in the low field region are integrated to 1, corresponding to the two protons on the amide bond at the triazole pyridine acylation and quinazoline amide, respectively; the interval of δ=9.46-7.28 corresponds to 11 protons in the aromatic region; the p peak at δ=5.31 is integrated to 1, corresponding to the protons on the chiral carbon; the s peak at δ=2.87 is integrated to 3, corresponding to the methyl at the 2-position of the quinazoline ring; the s peak at δ=2.10 is integrated to 1, corresponding to the proton of the cyclopropyl in the ortho position of the acyl group; the d peak at δ=1.56 is integrated to 3, corresponding to the methyl at the α-position of the chiral carbon; the d peak at δ=0.85 is integrated to 4, corresponding to the two methylene groups on the cyclopropyl.

[0120] Example 3: Synthesis of (R)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide

[0121] The raw materials are cyclopropylcarbonyl chloride and (R)-1-phenylethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0122] The product prepared in this embodiment is a white powder, and its structural formula is shown in Formula 3 above.

[0123] Physical, chemical and spectral data: Yield: 85%, melting point: >275°C, purity: 97.6%. 1H NMR (400MHz, DMSO-d6) δ11.12(s,1H),9.45(d,J=8.2Hz,1H),8.95(d,J=7.1Hz,1H), 8.71(s,1H),8.49(d,J=8.7Hz,1H),8.10(d,J=8.9Hz,1H),8.01(s,1H),7.50(d,J=7. 5Hz,2H),7.45-7.35(m,3H),7.30(t,J=7.3Hz,1H),5.30(p,J=7.2Hz,1H),2.87(s,3H ),2.10(s,1H),1.56(d,J=7.0Hz,3H),0.86(d,J=6.2Hz,4H).HRMS(m / z):calculated for C 28 H 26 O2N7[M+H] + 492.21425, found 492.21433.

[0124] Example 4: Synthesis of (S)-6-(2-isobutyramido-[1,2,4]triazolyl[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide

[0125] The raw materials are isobutyryl chloride and (S)-1-phenylethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0126] The product prepared in this embodiment is a white powder, and its structural formula is shown in Formula 4 above.

[0127] Physical, chemical and spectral data: white powder, yield: 78%, melting point: 241.3-242.6°C, purity: 97.2%. 1 HNMR(400MHz,DMSO-d6)δ10.78(s,1H),9.45(d,J=8.1Hz,1H),8.96(d,J=7.1Hz,1H),8 .71(d,J=2.1Hz,1H),8.49(dd,J=8.9,2.2Hz,1H),8.10(d,J=8.8Hz,1H),8.01(s,1H),7 .50(d,J=7.1Hz,2H),7.46-7.35(m,3H),7.30(t,J=7.3Hz,1H),5.31(p,J=7.2Hz,1H),2 .95-2.76(m,4H),1.56(d,J=7.0Hz,3H),1.13(d,J=6.8Hz,6H).HRMS(m / z):calculated forC 28 H 28O2N7[M+H] + 494.22990, found 494.23006.

[0128] Example 5: Synthesis of (S)-2-methyl-N-(1-phenylethyl)-6-(2-neoamido-[1,2,4]triazolo[1,5-a]pyridin-7-yl)quinazoline-4-carboxamide

[0129] The raw materials are pivaloyl chloride and (S)-1-phenylethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0130] The obtained product in this embodiment is a white powder, and its structural formula is shown in Formula 5 above.

[0131] Physical, chemical and spectral data: Yield: 85%, melting point: 251.1-251.9°C, purity: 99.5%. 1 H NMR (400MHz, DMSO-d6) δ10.30(s,1H),9.44(d,J=8.2Hz,1H),8.95(d,J=7.1Hz,1H) ,8.74(d,J=2.1Hz,1H),8.48(dd,J=8.9,2.1Hz,1H),8.10(d,J=8.9Hz,1H),8.00(s ,1H),7.51(d,J=7.6Hz,2H),7.44-7.35(m,3H),7.30(t,J=7.3Hz,1H),5.31(p,J=7 .2Hz,1H),2.87(s,3H),1.57(d,J=7.0Hz,3H),1.27(s,9H).HRMS(m / z):calculated for C 29 H 30 O2N7[M+H] + 508.24555, found 508.24574.

[0132] Example 6: Synthesis of (S)-2-methyl-6-(2-pentanamido-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-N-(1-phenylethyl)quinazoline-4-carboxamide

[0133] The raw materials are n-valeryl chloride and (S)-1-phenylethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0134] The product prepared in this embodiment is a white powder, and its structural formula is shown in Formula 6 above.

[0135] Physical, chemical and spectral data: Yield: 64%, melting point: >250°C, purity: 98.6%.1 H NMR (400MHz, DMSO-d6) δ10.79(s,1H),9.45(d,J=8.2Hz,1H),8.95(d,J=7.1Hz,1H),8.71(d,J =2.1Hz,1H),8.48(dd,J=8.9,2.1Hz,1H),8.10(d,J=8.8Hz,1H),8.00(s,1H),7.51(d,J=7.4H z,2H),7.45-7.34(m,3H),7.30(t,J=7.3Hz,1H),5.31(p,J=7.2Hz,1H),2.87(s,3H),2.48(s, 2H),1.66-1.53(m,5H),1.35(h,J=7.4Hz,2H),0.91(t,J=7.3Hz,3H).HRMS(m / z):calculated for C 29 H 30 O2N7[M+H] + 508.24555, found 508.24562.

[0136] Example 7: Synthesis of (S)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-(pyridin-2-yl)ethyl)quinazoline-4-carboxamide

[0137] The raw materials are cyclopropylcarbonyl chloride and (S)-1-(2-pyridyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0138] The obtained product in this embodiment is a white powder, and its structural formula is shown in Formula 7 above.

[0139] Physical, chemical and spectral data: Yield: 51%, melting point: >262°C, purity: 97.4%. 1H NMR (400MHz, DMSO-d6) δ11.12 (s, 1H), 9.51 (d, J = 7.7Hz, 1H), 9.09-8.92 (m, 2H), 8.60 (d ,J=4.9Hz,1H),8.51(d,J=8.7Hz,1H),8.17-8.02(m,2H),7.84(t,J=7.4Hz,1H),7.55(d ,J=7.9Hz,1H),7.47(d,J=7.1Hz,1H),7.39-7.27(m,1H),5.33(p,J=7.1Hz,1H),2.88(s ,3H),2.10(s,1H),1.57(d,J=7.0Hz,3H),0.86(d,J=5.6Hz,4H).HRMS(m / z):calculated for C 27 H 25 O2N8[M+H] + 493.20950, found 493.20966.

[0140] Example 8: Synthesis of (S)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-(pyridin-3-yl)ethyl)quinazoline-4-carboxamide

[0141] The raw materials are cyclopropylcarbonyl chloride and (S)-1-(3-pyridyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0142] The product prepared in this embodiment is a white powder, and its structural formula is shown in Formula 8 above.

[0143] Physical, chemical and spectral data: Yield: 48%, melting point: >272°C, purity: 98.1%. 1 H NMR (400MHz, DMSO-d6) δ11.12(s,1H),9.57(d,J=8.0Hz,1H),8.95(d,J=7.1Hz,1H ),8.74(dd,J=12.1,2.2Hz,2H),8.59-8.43(m,2H),8.10(d,J=8.8Hz,1H),8.01(s, 1H),7.92(d,J=8.0Hz,1H),7.49-7.35(m,2H),5.35(p,J=7.2Hz,1H),2.88(s,3H), 2.09(s,1H),1.60(d,J=7.0Hz,3H),0.86(d,J=6.0Hz,4H).HRMS(m / z):calculated for C 27 H 25O2N8[M+H] + 493.20950, found 493.20965.

[0144] Example 9: Synthesis of (S)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-(pyridin-4-yl)ethyl)quinazoline-4-carboxamide

[0145] The raw materials are cyclopropylcarbonyl chloride and (S)-1-(4-pyridyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0146] The product prepared in this embodiment is a white powder, and its structural formula is shown in Formula 9 above.

[0147] Physical, chemical and spectral data: Yield: 53%, melting point: >249°C, purity: 99.3%. 1 H NMR (400MHz, DMSO-d6) δ11.12(s,1H),9.58(d,J=7.8Hz,1H),8.95(d,J=7.1Hz,1H),8.82(d ,J=2.1Hz,1H),8.57(d,J=6.0Hz,2H),8.51(dd,J=9.0,2.1Hz,1H),8.12(d,J=8.9Hz,1H),8 .05(s,1H),7.49(d,J=6.1Hz,2H),7.43(dd,J=7.1,2.0Hz,1H),5.29(p,J=7.2Hz,1H),2.89 (s,3H),2.09(s,1H),1.57(d,J=7.1Hz,3H),0.85(d,J=6.1Hz,4H).HRMS(m / z):calculated forC 27 H 25 O2N8[M+H] + 493.20950, found 493.20964.

[0148] Example 10: Synthesis of (S)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-N-(1-(3-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide

[0149] The raw materials are cyclopropylcarbonyl chloride and (S)-1-(3-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0150] The powder prepared in this example is a light yellow powder, and the structural formula is shown in Formula 10 above.

[0151] Physical, chemical and spectral data: Yield: 68%, melting point: >266°C, purity: 97.2%. 1 H NMR (400MHz, DMSO-d6) δ11.12(s,1H),9.50(d,J=8.2Hz,1H),8.92(d,J=7.1Hz, 1H),8.74(s,1H),8.47(d,J=8.9Hz,1H),8.09(d,J=8.8Hz,1H),7.98(s,1H),7. 50-7.30(m,4H),7.13(t,J=8.5Hz,1H),5.33(p,J=7.2Hz,1H),2.88(s,3H),2.1 0(s,1H),1.57(d,J=7.1Hz,3H),0.87(d,J=4.3Hz,4H).HRMS(m / z):calculated for C 28 H 25 O2N7F[M+H] + 510.20483, found 510.20498.

[0152] Example 11: Synthesis of (S)-6-(2-(cyclopentylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide

[0153] The raw materials are cyclopentylcarbonyl chloride and (S)-1-phenylethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0154] The product prepared in this embodiment is a white powder, and the structural formula is shown in the above formula 11.

[0155] Physical and chemical data and spectral data: white powder, yield: 50%, melting point: >271°C, purity: 98.9%. 1H NMR (400MHz, DMSO-d6) δ10.80(s,1H),9.45(d,J=8.2Hz,1H),8.93(d,J=7.1Hz,1H),8.72( d,J=1.9Hz,1H),8.45(dd,J=8.9,2.2Hz,1H),8.07(d,J=8.8Hz,1H),7.96(d,J=1.5Hz,1H) ,7.52(d,J=7.3Hz,2H),7.40(t,J=7.5Hz,2H),7.37-7.26(m,2H),5.32(p,J=7.2Hz,1H),3 .03(s,1H),2.86(s,3H),1.97-1.84(m,2H),1.81-1.53(m,9H).HRMS(m / z):calculatedfor C 30 H 30 O2N7[M+H] + 520.24555, found 520.24572.

[0156] Example 12: Synthesis of (S)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-N-(1-(2-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide

[0157] The raw materials are cyclopropylcarbonyl chloride and (S)-1-(2-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0158] The powder prepared in this example is a light yellow powder, and the structural formula is shown in Formula 12 above.

[0159] Physical and chemical data and spectral data: light yellow powder, yield: 56%, melting point: >272°C, purity: 92.7%. 1H NMR (400MHz, DMSO-d6) δ11.11(s,1H),9.53(d,J=7.9Hz,1H),8.97(d,J=7.0Hz,1H),8.71 (d,J=2.1Hz,1H),8.51(dd,J=8.9,2.2Hz,1H),8.11(d,J=8.9Hz,1H),8.06-8.00(m,1H),7 .64-7.53(m,1H),7.44-7.33(m,2H),7.29-7.19(m,2H),5.53(p,J=7.2Hz,1H),2.88(s,3 H),2.09(s,1H),1.56(d,J=7.1Hz,3H),0.86(d,J=5.8Hz,4H).HRMS(m / z):calculatedfor C 28 H 25 O2N7F[M+H] + 510.20483, found 510.20496.

[0160] Example 13: Synthesis of (S)-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide

[0161] The raw materials are cyclopropylcarbonyl chloride and (S)-1-(4-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0162] The powder prepared in this example is a light yellow powder, and the structural formula is shown in Formula 13 above.

[0163] Physical, chemical and spectral data: Yield: 58%, melting point: >267°C, purity: 98.8%. 1 H NMR (400MHz, DMSO-d6) δ11.11(s,1H),9.45(d,J=8.2Hz,1H),8.93(d,J=7.1Hz,1H ),8.78(s,1H),8.49(d,J=8.9Hz,1H),8.17-7.97(m,2H),7.54(t,J=6.9Hz,2H),7 .39(d,J=7.2Hz,1H),7.21(t,J=8.7Hz,2H),5.31(p,J=6.5Hz,1H),2.87(s,3H),2 .10(s,1H),1.56(d,J=7.0Hz,3H),0.86(d,J=6.2Hz,4H).HRMS(m / z):calculated for C 28 H 25O2N7F[M+H] + 510.20483, found 510.20499.

[0164] Example 14: Synthesis of N-benzyl-6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methylquinazoline-4-carboxamide

[0165] The raw materials are cyclopropylcarbonyl chloride and benzylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0166] The product prepared in this embodiment is a white powder, and the structural formula is shown in Formula 14 above.

[0167] Physical, chemical and spectral data: Yield: 43%, melting point: >295°C, purity: 90.6%. 1 H NMR (400MHz, DMSO-d6) δ11.14(s,1H),9.65(t,J=6.2Hz,1H),9.07(d,J=2.1Hz ,1H),8.96(d,J=7.1Hz,1H),8.51(dd,J=8.9,2.2Hz,1H),8.18-8.03(m,2H),7. 49-7.42(m,3H),7.38(t,J=7.5Hz,2H),7.29(t,J=7.2Hz,1H),4.61(d,J=6.2Hz ,2H),2.87(s,3H),2.09(s,1H),0.85(d,J=6.2Hz,4H).HRMS(m / z):calculated for C 27 H 24 O2N7[M+H] + 478.19860, found 478.19875.

[0168] Example 15: Synthesis of 6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(4-(trifluoromethyl)benzyl)quinazoline-4-carboxamide

[0169] The raw materials are cyclopropylcarbonyl chloride and 4-trifluoromethylbenzylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0170] The product prepared in this embodiment is a white powder, and its structural formula is shown in Formula 15 above.

[0171] Physical, chemical and spectral data: Yield: 47%, melting point: >314°C, purity: 93.0%. 1H NMR (400MHz, DMSO-d6) δ11.13(s,1H),9.78(s,1H),9.19(s,1H),8.95(d,J=7.3Hz,1H),8.53(d,J=8.9Hz,1H),8.21-8.00(m,2H),7.75(d,J=8.1Hz ,2H),7.65(d,J=8.0Hz,2H),7.49(d,J=7.3Hz,1H),4.70(d,J=6.4Hz,2H),2.89(s,3H),2.09(s,1H),0.85(d,J=6.3Hz,4H).HRMS(m / z):calculated for C 28 H 23 O2N7F3[M+H] + 546.18598, found 546.18617.

[0172] Example 16: Synthesis of (S)-6-(2-(cyclohexylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide

[0173] The raw materials are cyclohexylcarbonyl chloride and (S)-1-phenylethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0174] The product prepared in this embodiment is a white powder, and its structural formula is shown in Formula 16 above.

[0175] Physical, chemical and spectral data: Yield: 58%, melting point: 278.2-278.8°C, purity: 97.0%. 1 H NMR (400MHz, DMSO-d6) δ10.72(s,1H),9.45(d,J=8.2Hz,1H),8.94(d,J=7.1Hz,1H),8.71(d,J=2.1Hz,1 H),8.48(dd,J=8.9,2.1Hz,1H),8.10(d,J=8.9Hz,1H),7.99(s,1H),7.50(d,J=7.4Hz,2H),7.46-7.33( m,3H),7.30(t,J=7.3Hz,1H),5.31(p,J=7.2Hz,1H),2.87(s,3H),2.57(s,1H),1.88-1.72(m,4H),1.66 (d,J=11.0Hz,1H),1.57(d,J=7.0Hz,3H),1.49-1.37(m,2H),1.35-1.20(m,3H).HRMS(m / z):calculated for C31 H 32 O2N7[M+H] + 534.26120, found 534.26132.

[0176] Example 17: Synthesis of (S)-6-(2-butylamino-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide

[0177] The raw materials are n-butyryl chloride and (S)-1-phenylethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0178] The product prepared in this embodiment is a white powder, and its structural formula is shown in Formula 17 above.

[0179] Physical, chemical and spectral data: Yield: 52%, melting point: >259°C, purity: 98.8%. 1 H NMR (400MHz, DMSO-d6) δ10.80(s,1H),9.45(d,J=8.2Hz,1H),8.95(d,J=7.1Hz,1H),8.71(s,1H),8.48(d,J=9.0Hz,1H),8.19-7.91(m, 2H),7.60-7.25(m,6H),5.50-5.14(m,1H),2.86(s,3H),2.46(s,2H),1.76-1.49(m,5H),0.94(t,J=7.3Hz,3H).HRMS(m / z):calculated for C 28 H 28 O2N7[M+H] + 494.22990, found 494.22997.

[0180] Example 18: Synthesis of 6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(4-(trifluoromethoxy)benzyl)quinazoline-4-carboxamide

[0181] The raw materials are cyclopropylcarbonyl chloride and 4-trifluoromethoxybenzylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0182] The product prepared in this embodiment is a white powder, and the structural formula is shown in Formula 18 above.

[0183] Physical, chemical and spectral data: Yield: 45%, melting point: >305°C, purity: 95.9%. 1H NMR (400MHz, DMSO-d6) δ11.11(s,1H),9.70(d,J=7.6Hz,1H),9.18(s,1H),8.96(s,1H),8.52(s,1H),8.11(s,2H ),7.60-7.46(m,3H),7.37(s,2H),4.63(s,2H),2.88(s,3H),2.09(s,1H),0.85(s,4H).HRMS(m / z):calculated for C 28 H 23 O3N7F3[M+H] + 562.18090, found 562.18108.

[0184] Example 19: Synthesis of (S)-6-(2-(cyclobutanecarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide

[0185] The raw materials are cyclobutylcarbonyl chloride and (S)-1-phenylethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0186] The product prepared in this embodiment is a white powder, and the structural formula is shown in Formula 19 above.

[0187] Physical, chemical and spectral data: Yield: 51%, melting point: 257.2-257.9°C, purity: 98.8%. 1 H NMR (400MHz, DMSO-d6) δ10.68(s,1H),9.46(d,J=8.1Hz,1H),9.07-8.90(m,1H),8.71(s,1H),8.50(d,J=8.8Hz,1H),8.20-7.96(m,2H),7.61-7.25( m,6H),5.31(s,1H),3.47(s,1H),2.87(s,3H),2.31-2.09(m,4H),2.02-1.89(m,1H),1.89-1.76(m,1H),1.68-1.45(m,3H).HRMS(m / z):calculated for C 29 H 28 O2N7[M+H] + 506.22990, found 506.23007.

[0188] Example 20: Synthesis of 6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(3-(trifluoromethyl)benzyl)quinazoline-4-carboxamide

[0189] The raw materials are cyclopropylcarbonyl chloride and 3-trifluoromethylbenzylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0190] The product prepared in this embodiment is a white powder, and its structural formula is shown in Formula 20 above.

[0191] Physical, chemical and spectral data: Yield: 46%, melting point: >299°C, purity: 94.0%. 1 H NMR (400MHz, DMSO-d6) δ11.11(s,1H),9.75(s,1H),9.11(s,1H),8.96(d,J=7.1Hz,1H),8.52(d,J=8.5Hz,1H),8.20-8.00(m,2H),7.82-7.72(m ,2H),7.71-7.57(m,2H),7.47(d,J=7.2Hz,1H),4.70(d,J=6.2Hz,2H),2.88(s,3H),2.09(s,1H),0.85(d,J=6.0Hz,4H).HRMS(m / z):calculated for C 28 H 23 O2N7F3[M+H] + 546.18598, found 546.18608.

[0192] Example 21: Synthesis of 6-(2-(cyclopropylcarboxamide)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-2-methyl-N-(3-(trifluoromethoxy)benzyl)quinazoline-4-carboxamide

[0193] The raw materials are cyclopropylcarbonyl chloride and 3-trifluoromethoxybenzylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0194] The product prepared in this embodiment is a white powder, and the structural formula is shown in Formula 21 above.

[0195] Physical, chemical and spectral data: Yield: 47%, melting point: >289°C, purity: 96.0%. 1H NMR (400MHz, DMSO-d6) δ11.11(s,1H),9.72(s,1H),9.21-8.87(m,2H),8.52(s,1H),8.24-7.97(m,2H),7. 57-7.34(m,4H),7.28(s,1H),4.65(s,2H),2.87(s,3H),2.07(s,1H),0.85(s,4H).HRMS(m / z):calculated for C 28 H 23 O3N7F3[M+H] + 562.18090, found 562.18096.

[0196] Example 22: Synthesis of (S)-2-methyl-6-(2-(3-oxocyclobutane-1-carboxamido)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)-N-(1-phenylethyl)quinazoline-4-carboxamide

[0197] The raw materials are 3-oxocyclobutanecarbonyl chloride and (S)-1-phenylethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0198] The powder prepared in this example is a light yellow powder, and the structural formula is shown in Formula 22 above.

[0199] Physical, chemical and spectral data: Yield: 54%, melting point: >210°C. 1 H NMR (400MHz, DMSO-d6) δ11.18(s,1H),9.46(d,J=8.3Hz,1H),8.97(t,J=6.5Hz,1H), 8.72(s,1H),8.49(d,J=9.0Hz,1H),8.06(m,2H),7.51(d,J=7.6Hz,2H),7.39(d,J=7 .2Hz,3H),7.30(t,J=7.0Hz,1H),5.30(q,J=7.2Hz,1H),3.57(s,1H),3.13-3.02(m, 2H),2.87(s,3H),2.44-2.21(m,2H),1.57(d,J=6.8Hz,3H).HRMS(m / z):calculated for C 29 H 24 O3N7[MH] - 518.19461, found 518.19502.

[0200] Example 23: Synthesis of (S)-N-(1-(4-fluorophenyl)ethyl)-2-methyl-6-(2-(3-oxocyclobutane-1-carboxamido)-[1,2,4]triazolo[1,5-a]pyridin-7-yl)quinazoline-4-carboxamide

[0201] The raw materials are 3-oxocyclobutanecarbonyl chloride and (S)-1-(4-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1) to (5) are the same as those in Example 1 (1) to (5).

[0202] The powder prepared in this example is a light yellow powder, and the structural formula is shown in Formula 23 above.

[0203] Physical, chemical and spectral data: Yield: 56%, melting point: >257°C. 1 H NMR (400MHz, DMSO-d6) δ11.18(s,1H),9.46(d,J=8.2Hz,1H),8.96(t,J=6.5Hz,1H),8.79(s,1H ),8.50(d,J=10.3Hz,1H),8.11(d,J=8.9Hz,1H),8.06(d,J=6.5Hz,1H),7.54(dd,J=8.4,5.6Hz, 2H),7.41(td,J=7.3,2.0Hz,1H),7.21(t,J=8.8Hz,2H),5.31(p,J=7.2Hz,1H),3.57(s,1H),3.3 3(s,1H),3.08(m,2H),2.87(s,3H),2.33(m,2H),1.56(d,J=7.0Hz,3H).HRMS(m / z):calculated forC 29 H 23 O3N7F[MH] - 536.18519, found 536.18557.

[0204] Example 24: Synthesis of (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide

[0205] The raw materials are cyclopropanecarboxylic acid, 2-amino-6-bromobenzothiazole, and (S)-1-phenylethylamine, which are reacted in equimolar amounts, and the remaining steps (1) to (5) are the same as those in Example 1. Among them, step (3) is modified as follows:

[0206] (3) Synthesis of N-(6-bromobenzo[d]thiazol-2-yl)cyclopropanecarboxamide:

[0207] 2-Amino-6-bromobenzothiazole (2.29 g, 10.0 mmol), small molecule carboxylic acid (11.0 mmol), dichloromethane (50 mL) and N-methylimidazole (2.05 g, 25.0 mmol) were added to a 100 mL round-bottom flask in sequence, mixed thoroughly, and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (3.65 g, 13 mmol) was slowly added under an ice-water bath. After the addition was complete, the ice bath was removed, the temperature was raised to room temperature, and the reaction was continued for another 2 h. The reaction was terminated after TLC monitoring of the reaction was complete. The reaction solution was extracted with ethyl acetate, the organic layer was dried, spin-dried, and purified by column chromatography to obtain the compound N-(6-bromobenzo[d]thiazol-2-yl)cyclopropanecarboxamide as a light pink powder with a yield of 85%.

[0208] The product prepared in this embodiment is a white powder, and its structural formula is shown in Formula 24 above.

[0209] Physicochemical and spectral data: Yield: 76%. 1 H NMR (400MHz, DMSO-d6) δ12.73(s,1H),9.44(d,J=8.2Hz,1H),8.55(d,J=2.1Hz,1H),8.39(dd,J =8.9,2.1Hz,1H),8.29(d,J=1.9Hz,1H),8.06(d,J=8.9Hz,1H),7.86(d,J=8.5Hz,1H),7.72(dd, J=8.4,1.9Hz,1H),7.50(d,J=7.4Hz,2H),7.39(t,J=7.5Hz,2H),7.30(t,J=7.3Hz,1H),5.30(p, J=7.2Hz,1H),2.85(s,3H),2.04(p,J=6.3Hz,1H),1.55(d,J=7.1Hz,3H),0.99(d,J=5.3Hz,4H).

[0210] Example 25: Synthesis of (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide

[0211] The raw materials are cyclopropanecarboxylic acid and (S)-1-(4-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0212] The product prepared in this embodiment is a white powder, and the structural formula is shown in Formula 25 above.

[0213] Physicochemical and spectral data: Yield: 75%. 1H NMR (400MHz, DMSO-d6) δ12.74(s,1H),9.44(s,1H),8.57(s,1H),8.36(d,J=18.3Hz,2H),8.06(d,J=8.7Hz,1H),7.85(d,J=8.2Hz,1H),7.69( d,J=8.2Hz,1H),7.59-7.42(m,2H),7.19(d,J=8.5Hz,2H),5.46-5.07(m,1H),2.85(s,3H),2.02(s,1H),1.54(s,3H),0.96(d,J=19.2Hz,4H).

[0214] Example 26: Synthesis of (S)-N-(1-(4-fluorophenyl)ethyl)-6-(2-isobutyramidobenzo[d]thiazol-6-yl)-2-methylquinazoline-4-carboxamide

[0215] The raw materials are isobutyric acid and (S)-1-(4-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0216] The powder prepared in this example is a light yellow powder, and the structural formula is shown in Formula 26 above.

[0217] Physicochemical and spectral data: Yield: 80%. 1 H NMR (400MHz, DMSO-d6) δ12.42(s,1H),9.45(d,J=8.2Hz,1H),8.58(s,1H),8.46-8.31(m,2H),8.07(d,J=8.9Hz,1H),7.85(d,J=8.6Hz,1H),7.70( d,J=8.6Hz,1H),7.60-7.49(m,2H),7.20(t,J=8.7Hz,2H),5.30(p,J=7.1 Hz,1H),2.92-2.77(m,4H),1.54(d,J=7.1Hz,3H),1.18(d,J=6.8Hz,6H).

[0218] Example 27: Synthesis of (S)-N-(1-(4-fluorophenyl)ethyl)-2-methyl-6-(2-pentanamidobenzo[d]thiazol-6-yl)quinazoline-4-carboxamide

[0219] The raw materials are n-pentanoic acid and (S)-1-(4-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0220] The product prepared in this example is a white powder, and its structural formula is shown in Formula 27 above.

[0221] Physicochemical and spectral data: Yield: 67%. 1 H NMR (400MHz, DMSO-d6) δ12.41(s,1H),9.45(d,J=8.2Hz,1H),8.58(d,J=2.1Hz,1H),8.46-8 .31(m,2H),8.07(d,J=8.8Hz,1H),7.84(d,J=8.4Hz,1H),7.69(dd,J=8.5,1.9Hz,1H),7.60 -7.48(m,2H),7.21(t,J=8.8Hz,2H),5.31(p,J=7.2Hz,1H),2.86(s,3H),2.56-2.52(m,2H) ,1.63(p,J=7.5Hz,2H),1.54(d,J=7.0Hz,3H),1.35(h,J=7.4Hz,2H),0.92(t,J=7.3Hz,3H).

[0222] Example 28: Synthesis of (S)-N-(1-(4-fluorophenyl)ethyl)-2-methyl-6-(2-(3-methylbutyramido)benzo[d]thiazol-6-yl)quinazoline-4-carboxamide

[0223] The raw materials are isovaleric acid and (S)-1-(4-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0224] The product prepared in this example is a yellow powder, and its structural formula is shown in Formula 28 above.

[0225] Physicochemical and spectral data: Yield: 74%. 1H NMR (400MHz, DMSO-d6) δ12.41(s,1H),9.45(d,J=8.2Hz,1H),8.58(d,J=2.1Hz,1H),8.40(dd,J =8.8,2.1Hz,1H),8.35(d,J=1.9Hz,1H),8.07(d,J=8.8Hz,1H),7.85(d,J=8.4Hz,1H),7.70(dd, J=8.5,1.9Hz,1H),7.58-7.50(m,2H),7.27-7.16(m,2H),5.30(p,J=7.2Hz,1H),2.86(s,3H),2. 41(d,J=7.2Hz,2H),2.14(dp,J=13.5,6.7Hz,1H),1.54(d,J=7.1Hz,3H),0.96(d,J=6.7Hz,6H).

[0226] Example 29: Synthesis of (S)-N-(1-(4-fluorophenyl)ethyl)-2-methyl-6-(2-pivalamidobenzo[d]thiazol-6-yl)quinazoline-4-carboxamide

[0227] The raw materials are pivalic acid and (S)-1-(4-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0228] The product prepared in this example is a yellow powder, and its structural formula is shown in Formula 29 above.

[0229] Physicochemical and spectral data: Yield: 78%. 1 H NMR (400MHz, DMSO-d6) δ12.16(s,1H),9.46(d,J=8.2Hz,1H),8.57(d,J=2.1Hz,1H) ,8.41(dd,J=8.9,2.1Hz,1H),8.36(d,J=1.9Hz,1H),8.07(d,J=8.8Hz,1H),7.85(d, J=8.4Hz,1H),7.70(dd,J=8.5,1.9Hz,1H),7.53(dd,J=8.5,5.6Hz,2H),7.20(t,J= 8.7Hz,2H),5.30(p,J=7.2Hz,1H),2.86(s,3H),1.54(d,J=7.0Hz,3H),1.30(s,9H).

[0230] Example 30: Synthesis of (S)-6-(2-(cyclobutanecarboxamido)benzo[d]thiazol-6-yl)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide

[0231] The raw materials are cyclobutanecarboxylic acid and (S)-1-(4-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0232] The product prepared in this embodiment is a white powder, and its structural formula is shown in Formula 30 above.

[0233] Physicochemical and spectral data: Yield: 75%. 1 H NMR (400MHz, DMSO-d6) δ12.31(s,1H),9.45(d,J=8.2Hz,1H),8.58(d,J=2.0Hz,1H), 8.48-8.25(m,2H),8.07(d,J=8.8Hz,1H),7.84(d,J=8.4Hz,1H),7.69(dd,J=8.5,1. 9Hz,1H),7.62-7.45(m,2H),7.33-7.10(m,2H),5.31(p,J=7.2Hz,1H),3.44(p,J=8. 3Hz,1H),2.86(s,3H),2.38-2.14(m,4H),2.07-1.80(m,2H),1.55(d,J=7.0Hz,3H).

[0234] Example 31: Synthesis of (S)-6-(2-(cyclopentanecarboxamido)benzo[d]thiazol-6-yl)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide

[0235] The raw materials are cyclopentanecarboxylic acid and (S)-1-(4-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0236] The powder prepared in this example is a yellow powder, and its structural formula is shown in Formula 31 above.

[0237] Physicochemical and spectral data: Yield: 66%. 1H NMR (400MHz, DMSO-d6) δ12.43(s,1H),9.46(d,J=8.2Hz,1H),8.57(d,J=2.1Hz,1 H),8.48-8.28(m,2H),8.07(d,J=8.9Hz,1H),7.84(d,J=8.4Hz,1H),7.70(dd,J=8 .5,1.9Hz,1H),7.63-7.44(m,2H),7.21(dd,J=10.1,7.7Hz,2H),5.30(p,J=7.2H z,1H),3.01(p,J=7.9Hz,1H),2.86(s,3H),2.00-1.88(m,2H),1.83-1.51(m,9H).

[0238] Example 32: Synthesis of (S)-6-(2-(cyclohexanecarboxamido)benzo[d]thiazol-6-yl)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide

[0239] The raw materials are cyclohexanecarboxylic acid and (S)-1-(4-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0240] The product prepared in this example is a yellow powder, and the structural formula is shown in Formula 32 above.

[0241] Physicochemical and spectral data: Yield: 81%. 1 H NMR (400MHz, DMSO-d6) δ12.37(s,1H),9.45(d,J=8.1Hz,1H),8.57(d,J=2.1Hz,1H),8.44-8.27(m,2H) ,8.07(d,J=8.9Hz,1H),7.84(d,J=8.4Hz,1H),7.69(dd,J=8.5,1.9Hz,1H),7.60-7.47(m,2H),7.32-7 .14(m,2H),5.30(p,J=7.3Hz,1H),2.85(s,3H),2.61-2.54(m,1H),1.88(d,J=12.7Hz,2H),1.77(d,J= 11.5Hz,2H),1.66(d,J=11.0Hz,1H),1.54(d,J=7.0Hz,3H),1.45(q,J=11.5Hz,2H),1.36-1.16(m,3H).

[0242] Example 33: Synthesis of (S)-6-(2-benzylaminobenzo[d]thiazol-6-yl)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide

[0243] The raw materials are benzoic acid and (S)-1-(4-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0244] The product prepared in this example is a yellow powder, and its structural formula is shown in Formula 33 above.

[0245] Physicochemical and spectral data: Yield: 70%. 1 H NMR (400MHz, DMSO-d6) δ12.98(s,1H),9.46(d,J=8.2Hz,1H),8.60(s,1H),8.39(d,J=7.8Hz,2H),8.17(d,J=7.6Hz,2H),8.07(d,J=8.8Hz,1H), 7.90(d,J=8.3Hz,1H),7.75-7.66(m,2H),7.64-7.51(m,4H),7.22(t,J=8.7Hz,2H),5.32(p,J=7.2Hz,1H),2.86(s,3H),1.56(d,J=7.1Hz,3H).

[0246] Example 34: Synthesis of (S)-6-(2-butyrylaminobenzo[d]thiazol-6-yl)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide

[0247] The raw materials are n-butyric acid and (S)-1-(4-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0248] The product prepared in this embodiment is a white powder, and the structural formula is shown in Formula 34 above.

[0249] Physicochemical and spectral data: Yield: 66%. 1H NMR(400MHz,DMSO-d6)δ12.41(s,1H),9.45(d,J=8.1Hz,1H),8.58(s,1H),8.44-8.31 (m,2H),8.07(d,J=8.8Hz,1H),7.84(d,J=8.4Hz,1H),7.69(dd,J=8.5,1.9Hz,1H),7.5 4(dd,J=8.4,5.5Hz,2H),7.20(t,J=8.8Hz,2H),5.31(p,J=7.2Hz,1H),2.86(s,3H),2 .53-2.51(m,2H),1.67(h,J=7.4Hz,2H),1.54(d,J=7.0Hz,3H),0.94(t,J=7.4Hz,3H).

[0250] Example 35: Synthesis of (S)-N-(1-(4-fluorophenyl)ethyl)-2-methyl-6-(2-propionamidobenzo[d]thiazol-6-yl)quinazoline-4-carboxamide

[0251] The raw materials are propionic acid and (S)-1-(4-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0252] The product prepared in this embodiment is a white powder, and the structural formula is shown in Formula 35 above.

[0253] Physicochemical and spectral data: Yield: 66%. 1 H NMR (400MHz, DMSO-d6) δ12.39 (s, 1H), 9.45 (d, J = 8.2Hz, 1H), 8.58 (s, 1H), 8.4 2-8.30(m,2H),8.06(d,J=8.8Hz,1H),7.84(d,J=8.4Hz,1H),7.69(d,J=8.5Hz, 1H),7.54(dd,J=8.4,5.5Hz,2H),7.21(t,J=8.7Hz,2H),5.31(p,J=7.2Hz,1H) ,2.86(s,3H),2.59-2.52(m,2H),1.55(d,J=7.0Hz,3H),1.14(t,J=7.5Hz,3H).

[0254] Example 36: Synthesis of (S)-6-(2-acetylaminobenzo[d]thiazol-6-yl)-N-(1-(4-fluorophenyl)ethyl)-2-methylquinazoline-4-carboxamide

[0255] The raw materials are acetic acid and (S)-1-(4-fluorophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0256] The product prepared in this embodiment is a white powder, and the structural formula is shown in Formula 36 above.

[0257] Physicochemical and spectral data: Yield: 69%. 1 H NMR (400MHz, DMSO-d6) δ12.43(s,1H),9.45(d,J=8.2Hz,1H),8.58(d,J=2.1Hz,1H),8.46-8.30(m,2H),8.07(d,J=8.8Hz,1H),7.85(d,J=8.4Hz,1 H),7.69(dd,J=8.4,1.9Hz,1H),7.61-7.50(m,2H),7.20(t,J=8.8Hz,2H) ,5.31(p,J=7.3Hz,1H),2.86(s,3H),2.24(s,3H),1.55(d,J=7.0Hz,3H).

[0258] Example 37: Synthesis of N-benzyl-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methylquinazoline-4-carboxamide

[0259] The raw materials are cyclopropanecarboxylic acid and benzylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0260] The product prepared in this embodiment is a white powder, and its structural formula is shown in Formula 37 above.

[0261] Physicochemical and spectral data: Yield: 62%. 1H NMR (400MHz, DMSO-d6) δ12.73(s,1H),9.60(t,J=6.3Hz,1H),8.93(d,J=2.0Hz,1H ),8.45-8.31(m,2H),8.06(d,J=8.8Hz,1H),7.86(d,J=8.4Hz,1H),7.77(dd,J=8. 5,1.9Hz,1H),7.45(d,J=7.4Hz,2H),7.38(t,J=7.5Hz,2H),7.29(t,J=7.2Hz,1H) ,4.61(d,J=6.2Hz,2H),2.85(s,3H),2.04(p,J=6.2Hz,1H),0.99(d,J=6.1Hz,4H).

[0262] Example 38: Synthesis of (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(1-phenylpropyl)quinazoline-4-carboxamide

[0263] The raw materials are cyclopropanecarboxylic acid and (S)-1-phenylpropylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0264] The product prepared in this embodiment is a white powder, and the structural formula is shown in Formula 38 above.

[0265] Physicochemical and spectral data: Yield: 71%. 1 H NMR (400MHz, DMSO-d6) δ12.74(s,1H),9.40(d,J=8.5Hz,1H),8.46(d,J=2.0Hz,1H),8.39(dd,J= 8.8,2.1Hz,1H),8.26(s,1H),8.06(d,J=8.8Hz,1H),7.85(d,J=8.4Hz,1H),7.69(dd,J=8.5,1.9 Hz,1H),7.48(d,J=7.4Hz,2H),7.39(t,J=7.5Hz,2H),7.30(t,J=7.3Hz,1H),5.05(q,J=7.9Hz,1 H),2.85(s,3H),2.04(p,J=6.2Hz,1H),1.88(ddp,J=20.7,13.7,7.1Hz,2H),1.05-0.91(m,7H).

[0266] Example 39: Synthesis of 6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-phenylquinazoline-4-carboxamide

[0267] The raw materials are cyclopropanecarboxylic acid and aniline, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0268] The product prepared in this example is a yellow powder, and its structural formula is shown in Formula 39 above.

[0269] Physicochemical and spectral data: Yield: 76%. 1 H NMR (400MHz, DMSO-d6) δ12.72(s,1H),10.93(s,1H),8.87(d,J=2.1Hz,1H),8.48-8.37(m,2H),8.10(d,J=8.9Hz,1H),7.93-7. 82(m,4H),7.43(dd,J=8.5,7.4Hz,2H),7.25-7.16(m,1H),2.91(s,3H),2.03(ddd,J=9.8,8.0,5.7Hz,1H),1.05-0.93(m,4H).

[0270] Example 40: Synthesis of 6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-phenethylquinazoline-4-carboxamide

[0271] The raw materials are cyclopropanecarboxylic acid and phenethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0272] The product prepared in this embodiment is a yellow powder, and the structural formula is shown in the above formula 40.

[0273] Physicochemical and spectral data: Yield: 65%. 1 H NMR (400MHz, DMSO-d6) δ12.74(s,1H),9.11(t,J=5.9Hz,1H),8.88(d,J=2.1Hz, 1H),8.45-8.28(m,2H),8.04(d,J=8.8Hz,1H),7.88(d,J=8.4Hz,1H),7.79(dd, J=8.4,1.9Hz,1H),7.34-7.24(m,4H),7.19-7.11(m,1H),3.65(q,J=6.6Hz,2H) ,2.94(t,J=7.4Hz,2H),2.84(s,3H),2.05(p,J=6.2Hz,1H),1.14-0.90(m,4H).

[0274] Example 41: Synthesis of 6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(tetrahydro-2H-pyran-4-yl)quinazoline-4-carboxamide

[0275] The raw materials are cyclopropanecarboxylic acid and 4-aminotetrahydropyran, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0276] The powder prepared in this example is a light yellow powder, and the structural formula is shown in Formula 41 above.

[0277] Physicochemical and spectral data: Yield: 78%. 1 H NMR (400MHz, DMSO-d6) δ12.73(s,1H),8.98(d,J=8.0Hz,1H),8.83(d,J=2.0Hz,1H),8.40(dt,J= 5.5,2.9Hz,2H),8.07(d,J=8.9Hz,1H),7.88(d,J=8.5Hz,1H),7.82(dd,J=8.5,1.9Hz,1H),4.16 (ddp,J=11.5,8.3,4.2Hz,1H),3.98-3.86(m,2H),3.44(td,J=11.7,2.1Hz,2H),2.85(s,3H),2. 10-1.99(m,1H),1.87(dt,J=12.8,2.7Hz,2H),1.66(qd,J=11.9,4.4Hz,2H),1.09-0.89(m,4H).

[0278] Example 42: Synthesis of (S)-N-(1-cyclohexylethyl)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methylquinazoline-4-carboxamide

[0279] The raw materials are cyclopropanecarboxylic acid and (S)-1-cyclohexylethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0280] The product prepared in this example is a yellow powder, and the structural formula is shown in Formula 42 above.

[0281] Physicochemical and spectral data: Yield: 79%. 1H NMR (400MHz, DMSO-d6) δ12.74(s,1H),8.82-8.66(m,2H),8.45-8.34(m,2H),8.06(d,J=8.9Hz,1H),7.87(d,J=8.4Hz,1H),7.80(dd,J=8.4,1.9H z,1H),3.99(dt,J=8.7,6.8Hz,1H),2.84(s,3H),2.09-1.99(m,1H),1.9 0-1.68(m,4H),1.66-1.42(m,2H),1.27-1.03(m,8H),1.02-0.96(m,4H).

[0282] Example 43: Synthesis of (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(1-(p-tolyl)ethyl)quinazoline-4-carboxamide

[0283] The raw materials are cyclopropanecarboxylic acid and (S)-1-(4-methylphenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0284] The product prepared in this example is a yellow powder, and its structural formula is shown in Formula 43 above.

[0285] Physicochemical and spectral data: Yield: 62%. 1 H NMR (400MHz, DMSO-d6) δ12.74(s,1H),9.38(d,J=8.2Hz,1H),8.55(d,J=2.1Hz,1H),8.39( dd,J=8.9,2.1Hz,1H),8.32(d,J=1.9Hz,1H),8.06(d,J=8.9Hz,1H),7.85(d,J=8.4Hz,1H), 7.71(dd,J=8.5,1.9Hz,1H),7.42-7.34(m,2H),7.18(d,J=7.8Hz,2H),5.26(p,J=7.2Hz,1 H),2.85(s,3H),2.29(s,3H),2.10-1.98(m,1H),1.52(d,J=7.0Hz,3H),1.07-0.92(m,4H).

[0286] Example 44: Synthesis of (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(1-(pyridin-4-yl)ethyl)quinazoline-4-carboxamide

[0287] The raw materials are cyclopropanecarboxylic acid and (S)-1-(pyridin-4-yl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as the modified (3) of Example 24.

[0288] The product prepared in this example is a yellow powder, and the structural formula is shown in Formula 44 above.

[0289] Physicochemical and spectral data: Yield: 75%. 1 H NMR (400MHz, DMSO-d6) δ12.74(s,1H),9.57(d,J=7.9Hz,1H),8.62(d,J=2.1Hz,1H),8. 60-8.55(m,2H),8.41(dd,J=8.9,2.1Hz,1H),8.34(d,J=1.9Hz,1H),8.08(d,J=8.8Hz,1 H),7.86(d,J=8.4Hz,1H),7.73(dd,J=8.4,2.0Hz,1H),7.53-7.45(m,2H),5.28(p,J=7. 1Hz, 1H), 2.88 (s, 3H), 2.04 (p, J = 6.4Hz, 1H), 1.55 (d, J = 7.1Hz, 3H), 1.13-0.88 (m, 4H).

[0290] Example 45: Synthesis of (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-N-(1-(4-methoxyphenyl)ethyl)-2-methylquinazoline-4-carboxamide

[0291] The raw materials are cyclopropanecarboxylic acid and (S)-1-(4-methoxyphenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0292] The product prepared in this example is a yellow powder, and the structural formula is shown in Formula 45 above.

[0293] Physicochemical and spectral data: Yield: 79%. 1H NMR (400MHz, DMSO-d6) δ12.75 (s, 1H), 9.35 (d, J = 8.3Hz, 1H), 8.56 (s, 1H), 8.43-8 .28(m,2H),8.05(d,J=8.8Hz,1H),7.84(d,J=8.4Hz,1H),7.67(dd,J=8.4,1.8Hz, 1H),7.42(d,J=8.3Hz,2H),6.94(d,J=8.2Hz,2H),5.26(p,J=7.2Hz,1H),3.74(s, 3H), 2.84 (s, 3H), 2.04 (p, J = 6.3Hz, 1H), 1.53 (d, J = 7.0Hz, 3H), 1.07-0.94 (m, 4H).

[0294] Example 46: Synthesis of (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(2-methyl-1-phenylpropyl)quinazoline-4-carboxamide

[0295] The raw materials are cyclopropanecarboxylic acid and (S)-2-methyl-1-phenylpropan-1-amine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4) and (5) are the same as (1), (2), (4) and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0296] The product prepared in this example is a yellow powder, and its structural formula is shown in Formula 46 above.

[0297] Physicochemical and spectral data: Yield: 71%. 1 H NMR (400MHz, DMSO-d6) δ12.75(s,1H),9.41(d,J=9.0Hz,1H),8.37(d,J=10.9Hz,2H),8.21(s,1H) ,8.05(d,J=8.6Hz,1H),7.84(d,J=8.4Hz,1H),7.65(d,J=9.3Hz,1H),7.47(d,J=7.7Hz,2H),7.39( t,J=7.5Hz,2H),7.31(t,J=6.8Hz,1H),4.86(t,J=9.0Hz,1H),2.85(s,3H),2.17(dp,J=8.8,6.6H z,1H),2.05(p,J=6.2Hz,1H),1.07(d,J=7.0Hz,3H),1.00(d,J=6.0Hz,4H),0.81(d,J=6.6Hz,3H).

[0298] Example 47: Synthesis of (R)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide

[0299] The raw materials are cyclopropanecarboxylic acid and (R)-1-phenylethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0300] The product prepared in this embodiment is a white powder, and its structural formula is shown in Formula 47 above.

[0301] Physicochemical and spectral data: Yield: 63%. 1 H NMR (400MHz, DMSO-d6) δ12.74(s,1H),9.45(d,J=8.2Hz,1H),8.54(d,J=2.1Hz,1H),8.40(dd,J= 8.9,2.1Hz,1H),8.30(d,J=1.9Hz,1H),8.07(d,J=8.8Hz,1H),7.86(d,J=8.4Hz,1H),7.73(dd,J =8.5,1.9Hz,1H),7.55-7.46(m,2H),7.39(dd,J=8.4,6.8Hz,2H),7.33-7.27(m,1H),5.29(p,J= 7.2Hz,1H),2.85(s,3H),2.04(dt,J=12.5,6.2Hz,1H),1.55(d,J=7.1Hz,3H),1.04-0.95(m,4H).

[0302] Example 48: Synthesis of (S)-6-(2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)-2-methyl-N-(1-(4-nitrophenyl)ethyl)quinazoline-4-carboxamide

[0303] The raw materials are cyclopropanecarboxylic acid and (S)-1-(4-nitrophenyl)ethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), (4), and (5) are the same as (1), (2), (4), and (5) of Example 1, and step (3) is the same as (3) after modification of Example 24.

[0304] The product prepared in this example is a yellow powder, and its structural formula is shown in Formula 48 above.

[0305] Physicochemical and spectral data: Yield: 74%. 1H NMR (400MHz, DMSO-d6) δ12.74(s,1H),9.64(d,J=7.8Hz,1H),8.61(d,J=2.1Hz,1H),8. 40(dd,J=8.9,2.1Hz,1H),8.37(d,J=1.9Hz,1H),8.29-8.23(m,2H),8.08(d,J=8.8Hz, 1H),7.82(d,J=8.5Hz,1H),7.80-7.74(m,2H),7.69(dd,J=8.4,1.9Hz,1H),5.40(p,J= 7.1Hz,1H),2.88(s,3H),2.15-1.97(m,1H),1.58(d,J=7.0Hz,3H),1.02-0.94(m,4H).

[0306] Example 49: Synthesis of (S)-6-((2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)methyl)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide

[0307] The raw materials are cyclopropanecarboxylic acid, 6-(bromomethyl)benzo[d]thiazol-2-amine, and (S)-1-phenylethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), and (4) are the same as (1), (2), and (5) of Example 1, and step (3) is the same as (3) of Example 24 after modification.

[0308] The structural formula prepared in this example is shown in Formula 49 above.

[0309] Example 50: Synthesis of (S)-6-((2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)amino)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide

[0310] The raw materials are cyclopropanecarboxylic acid, 6-nitrobenzo[d]thiazol-2-amine, and (S)-1-phenylethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), and (5) are the same as those in Example 1 (1), (2), and (5), and step (3) is the same as the modified step (3) in Example 24. Among them, step (4) is modified as follows:

[0311] (4) Synthesis of N-(6-aminobenzo[d]thiazol-2-yl)cyclopropanecarboxamide:

[0312] N-(6-nitrobenzo[d]thiazol-2-yl)cyclopropanecarboxamide and stannous chloride were reacted in a methanol / chloroform mixed solvent under reflux or at room temperature overnight. After the reaction was completed, the solvent was partially dried by TLC monitoring, the pH was adjusted to alkaline, the reaction solution was extracted with dichloromethane, the organic layer was dried, and the compound N-(6-aminobenzo[d]thiazol-2-yl)cyclopropanecarboxamide was obtained by drying, which can be directly used in the next step without further purification.

[0313] The structural formula prepared in this example is shown in Formula 50 above.

[0314] Example 51: Synthesis of (S)-6-((2-(cyclopropanecarboxamido)benzo[d]thiazol-6-yl)oxy)-2-methyl-N-(1-phenylethyl)quinazoline-4-carboxamide

[0315] The raw materials are cyclopropanecarboxylic acid, 6-hydroxybenzo[d]thiazol-2-amine, and (S)-1-phenylethylamine, which are reacted in equimolar amounts. The remaining steps (1), (2), and (4) are the same as (1), (2), and (5) of Example 1, and step (3) is the same as the modified (3) of Example 24. Among them, the condensation reaction of step (3) needs to protect the hydroxyl group with tert-butyldimethylsilyl chloride before the reaction, and tetrabutylammonium fluoride is used to remove the protecting group after the condensation, and the modification is as follows:

[0316] Protection of the hydroxyl group in 2-aminobenzo[d]thiazol-6-ol:

[0317] 2-aminobenzo[d]thiazole-6-ol, tert-butyldimethylsilyl chloride and imidazole were stirred in N,N-dimethylformamide overnight. After the reaction was completed, the product of the protective agent was extracted with ethyl acetate under TLC monitoring. The organic layer was dried and spin-dried to obtain the hydroxyl-protected compound 6-((tert-butyldimethylsilyl)oxy)benzo[d]thiazole-2-amino.

[0318] Removal of hydroxyl protecting group:

[0319] N-(6-((tert-butyldimethylsilyl)oxy)benzo[d]thiazol-2-yl)cyclopropanecarboxamide was added to a tetrabutylammonium fluoride tetrahydrofuran solution and stirred overnight at room temperature. After the reaction was completed by TLC monitoring, part of the tetrahydrofuran was dried and the deprotected hydroxyl product was extracted with dichloromethane. The organic layer was dried and dried to obtain the deprotected compound N-(6-hydroxybenzo[d]thiazol-2-yl)cyclopropanecarboxamide.

[0320] The structural formula prepared in this example is shown in Formula 51 above.

[0321] Example 52: Inhibitory effect of compounds 1-23 on RIPK1

[0322] The inhibitory effect of the compounds on RIPK1 and RIPK3 was determined by using the ADP-Glo ​​Kinase Assay Kit and the selectivity index was calculated. The selected RIPK1 positive control was GSK2982772 and the RIPK3 positive control was GSK-872. Based on this, the inhibitory ability of compounds 1-23 on RIPK1 was tested and the measured IC 50 Compounds with a value less than 200 nM were further tested for their inhibitory ability against RIPK3, and the selectivity index (SI) was calculated.

[0323] According to the instructions of the ADP-Glo ​​Kinase Assay Kit (Cat. No.: #V9102), read the fluorescence intensity and calculate the inhibition rate. The fluorescence intensity of the compound well is recorded as Lum compound The fluorescence intensity of the control well is recorded as Lum vehicle The fluorescence intensity of the blank control well is recorded as Lum blank , the inhibition rate calculation formula is as follows:

[0324] Inhibition rate = (Lum vehicle -Lum compound ) / (Lum vehicle -Lum blank )×100%.

[0325] The structural formula of compound 1-23 is shown in formula (III), and the research results are shown in Table 1.

[0326]

[0327] Nonlinear regression fitting was performed based on the scatter plots from GraphPad Prizm and IC was calculated 50 value.

[0328] The results of the study are as shown in Table 1. ① Positive control and IC reported in the literature 50 The values ​​are close, indicating that the modeling is successful and the experimental results are credible; ② The IC values ​​of compounds 2, 7, 9, 11, 12, and 14 for RIPK1 50 Value less than 200nM (IC 50 =122.4~188.2nM). Further experimental results showed that the above six compounds had no inhibitory effect on RIPK3 at the maximum test concentration of 10μM, and the calculated SI values ​​were all greater than 50. Among them, the SI values ​​of compounds 2 and 15 were close to or greater than 80, proving that compounds 1 and 15 had good selective inhibitory effects on RIPK1.

[0329] Example 53: Inhibitory effect of compounds 24-48 on RIPK1

[0330] The inhibitory effect of the compound on RIPK1 and RIPK3 was determined by using ADP-Glo ​​Kinase Assay Kit and the selectivity index was calculated. The implementation scheme is the same as Example 52. ADP-Glo ​​Kinase Assay Kit (Cat. No.: #V9102).

[0331] The structural formula of compound 24-48 is shown in formula (IV), and the research results are shown in Table 2.

[0332]

[0333] The results of the study are as follows: As shown in Table 2, ① positive control and IC reported in the literature 50 The values ​​are close, indicating that the modeling is successful and the experimental results are credible; ② The IC values ​​of compounds 24, 25, 27, 28, 30, 31, 32, 33, 34, 35, 36, 38, 43, 45, 48 for RIPK1 50 Further experimental results showed that the SI values ​​of compounds 25, 27, 28, 30, 31, 32, 33, 34, 35, 36, 38, 43, and 45 for RIPK3 were all greater than 75, proving that these compounds all had good selective inhibitory effects on RIPK1.

[0334] Example 54: Protective effect of compound 1-23 on HT-29 cells

[0335] (1) Cell culture

[0336] The HT-29 cells used in the experiment were cultured in RPMI 1640 medium containing 10% FBS; the HT-22 cells were cultured in DMEM high-glucose medium containing 10% FBS and grown at 37° C. in a 5% CO 2 environment. All cells were in the logarithmic growth phase.

[0337] (2) TSZ method to establish a programmed cell necrosis model

[0338] HT-29 cells were digested with trypsin, centrifuged, and counted at 1×10 4 The cells were evenly inoculated in a 96-well plate at a density of 10 cells / well. After the cells were completely attached to the wall, the culture medium was replaced and TSZ modeling reagent was added to make the concentrations in the wells 10 ng / mL TNF-α, 500 nM Smac-mimetic BV6, and 20 μM Z-VAD (OMe) -FMK, respectively. Each well contained 100 μL of culture medium. After 24 hours, CCK-8 detection reagent was added and the OD value was detected after incubation in the incubator for 2-3 hours.

[0339] (3) CCK-8 cytotoxicity assay

[0340] The cell plating operation is as in step (2); the culture medium containing different concentrations of the compound to be tested prepared in advance is added to the wells, and after continuing to culture for 24 hours, the cell survival rate is determined by the CCK-8 method.

[0341] (4) CCK-8 assay to detect cell viability and calculate EC 50 value

[0342] The cell plating operation is as in step (2). The prepared culture medium containing different concentrations of the compound to be tested is added to the wells. After pre-incubation for 30 minutes, TSZ is added. After 24 hours, the cell viability is determined using the CCK-8 method. Nonlinear regression fitting is performed based on the scatter plots of GraphPadPrizm, and the EC 50 value.

[0343] The research results show that: as shown in Table 1, Figure 1-Figure 4 As shown, compounds 2, 10, 12, 13, 15, and 23 performed well, with their EC 50 The values ​​were in the range of 66 to 103 nM. Compounds 6-10, 14, 15, and 19 all showed toxicity to the normal growth of HT-29 cells. Compound 2 had a better protective effect and lower cytotoxicity.

[0344] Example 55: Protective effect of compounds 23-48 on HT-29 cells

[0345] The implementation scheme is the same as Example 24. HT-22 cells were used to induce programmed necrosis model by TSZ method, and CCK-8 cytotoxicity detection and cell survival rate were performed to evaluate the toxicity of compounds 23-48 and their protective effects on HT-29 cells.

[0346] The results show that, as shown in Table 2, the EC values ​​of compounds 24, 25, 35, 38, 42, 43, 44, 46, and 48 50 The value was less than 35 nM, which had a good protective effect on HT-29 cells.

[0347] Example 56: Protective effect of compound 2 on HT-22, HT-29, L929 and U937 cells

[0348] HT-22, HT-29, L929 and U937 cells in the logarithmic growth phase were plated in 24-well plates and allowed to adhere overnight in a 37°C incubator containing 5% CO2. After pre-incubation with compound 2 for 30 minutes, HT-22, HT-29 and U937 cells were treated with TSZ for 6 hours; L929 cells were treated with TZ for 4 hours, and the cell morphology was observed under a microscope and photographed.

[0349] The research results are as follows Figure 2-Figure 4As shown, the EC of compound 2 on HT-22, HT-29, L929 and U937 cells 50 The EC values ​​of L929 cells were 38.45±2.96nM, 71.61±3.32nM, 24.84±4.14nM and 152.9±34.56nM, respectively. 50 =24.84±4.14 nM, EC of HT-22 cells 50 =38.45±2.96 nM; Compound 2 effectively restored cell viability in TSZ-induced necrosis models of different cell lines.

[0350] Example 57: Fluorescence staining experiment of compound 2 protecting cells

[0351] HT-29 or L929 cells were digested with trypsin, centrifuged and counted, and then seeded in a 24-well plate at a certain density. After the cells adhered to the wall and grew, new culture medium with 1, 10, and 100 nM compound concentrations was added to the drug-treated group, and an equal volume of RPMI 1640 complete culture medium containing 10% FBS was added to the control group and the model group. After pre-incubation for 30 minutes, TSZ was added to the drug-treated group and the model group for a certain period of time except for the control group. The culture medium was discarded, and Hoechst33342 and PI dyes were added to the cells after rinsing with PBS, and then placed in a 37°C constant temperature incubator and incubated in the dark for 30 minutes. After the incubation, the cells were rinsed again with PBS, and new blank culture medium was added. The staining of Hoechst33342 and PI in the cells was observed under a fluorescence microscope and photographed.

[0352] The research results show that: Figure 5 , Figure 6 As shown in the figure, compared with the control group, the blue fluorescence of the TSZ modeling group was enhanced, but after administration, the blue fluorescence between the dose groups was weakened but the difference was not obvious. This is because the membrane permeability increases after the cells undergo programmed necrosis, resulting in more Hoechst 33342 dye entering the cell nucleus to color DNA. The red fluorescence of PI staining is more obvious in the TSZ group than in the control group, and the red fluorescence of L929 cells almost disappears after administration; the red fluorescence of HT-29 cells also weakens in a dose-dependent manner after administration. The above can show that compound 2 can play a protective role by protecting cells from programmed necrosis.

[0353] Example 58: Protective effect of compound 2 on MCAO rats

[0354] 1. Experimental methods:

[0355] About 72 male SD rats were randomly divided into 6 groups according to body weight: A. sham operation group, B. model control group, C. low-dose group of compound 2 (0.5 mg / kg), D. medium-dose group of compound 2 (1.5 mg / kg), E. high-dose group of compound 2 (4.5 mg / kg), and F. positive control group. There were 12 rats in each group. Each group was first intravenously injected once 0.5 h before modeling, and then once again after reperfusion, for a total of 2 times. Except for the control group, the remaining groups were prepared with MCAO models. Neurological scores were performed 24 h later, and blood was taken after the scoring to detect the levels of SOD, MDA, IL-1β, and TNF-α. Brain sections were taken for TTC staining to observe cerebral ischemia and infarction.

[0356] 2. Research results:

[0357] ① Zea Longa neurological score: The neurological score of the animals was performed 24 hours after surgery according to the Zea Longa 4-point scoring standard. The results showed that compared with the model control group, the neurological scores of the animals in each drug-treated group tended to be alleviated, among which the high-dose group of the test sample and the positive control group were significantly reduced (p < 0.05 or p < 0.01), see Table 3, Figure 7 .

[0358] ②mNSS neurological score: The neurological score of the animals was evaluated 24 hours after surgery according to the 18-point mNSS scoring standard. The results showed that the neurological scores of the animals in each drug-treated group were significantly reduced compared with those in the model control group (p < 0.05 or p < 0.001), as shown in Table 3. Figure 7 .

[0359] ③ Percentage of cerebral infarction volume: Compared with the model control group, the percentage of cerebral infarction volume in each drug-treated group showed a decreasing trend, among which the percentage of cerebral infarction volume in the medium- and high-dose groups was significantly reduced (p < 0.01 or p < 0.001), as shown in Table 4. Figure 8 .

[0360] ④ Results of serum antioxidant indexes: Compared with the model control group, the serum SOD content of animals in each drug-treated group tended to increase, and the positive control group increased significantly (p < 0.05); compared with the model control group, the serum MDA content of animals in each drug-treated group tended to decrease, and the medium- and high-dose groups of the tested samples and the positive control group decreased significantly (p < 0.05 or p < 0.01); compared with the model control group, the serum TNF-ɑ and IL-1β content of animals in each drug-treated group tended to decrease, and the medium- and high-dose groups of the tested samples and the positive control group decreased significantly (p < 0.05 or p < 0.01), see Table 5, Fig. 9 .

[0361] Under the experimental conditions, the compound has a significant improvement effect on acute cerebral ischemia-reperfusion injury in rats, can significantly reduce the neurobehavioral score of cerebral ischemia injury, and reduce the volume of cerebral infarction. Its mechanism of action may be related to its antioxidant and anti-inflammatory effects.

[0362] Example 59: Acute toxicity of compound 2

[0363] SPF-grade male Kunming mice weighing 18-22 g were purchased from the Experimental Animal Center of the School of Pharmacy, Sun Yat-sen University. The experimental animal certificate number is: 44008500033846, and the experimental animal use license number is: SYXK (Guangdong) 2021-0029.

[0364] The maximum dissolution concentration of the drug solution was prepared to be 2 mg / ml, and a larger dose of 0.4 mL / 10 g was given by intraperitoneal injection to mice. The high, medium and low doses were set downward by a coefficient of 0.7, which were 160, 112 and 78.4 mg / kg respectively.

[0365] 16 mice were randomly divided into 4 groups according to their weight, 4 mice in each group, including high-dose group of compound 2 (160 mg / kg), medium-dose group of compound 2 (112 mg / kg), low-dose group of compound 2 (78.4 mg / kg), and vehicle control group. The mice were intraperitoneally injected (2 times, 1 hour apart), and the animals were observed immediately after administration for 12 hours and then for 7 consecutive days to record the general condition and body weight of the mice.

[0366] The results of the study showed that, as shown in Table 6, compared with the vehicle control group, the body weight of animals in the high and medium dose groups (160 and 112 mg / kg) of compound 2 showed a slight decrease within 7 days after administration, but no statistical difference was observed. Compound 2 did not show any significant abnormality in other general conditions of the animals in each group, such as diet, appearance, behavior, secretions, excretions, etc.

[0367] Under the experimental conditions, compound 2 did not produce obvious acute toxicity to Kunming mice when administered intraperitoneally within the maximum concentration and maximum dosage range of 160 mg / kg. Only a trend of weight loss was observed, and no obvious abnormal changes in state or death were observed. It is inferred that the LD 50 The dose is above 160mg / kg.

[0368] Example 60: Pharmacokinetics of Compound 2

[0369] SPF-grade SD male rats weighing 180-200 g were purchased from the Experimental Animal Center of the School of Pharmacy, Sun Yat-sen University. The experimental animal qualification certificate number is: No. 44007200122654; the experimental animal use license number is: SYXK (Guangdong) 2016-0029.

[0370] Rats were randomly divided into 2 groups, 6 rats in each group, including intravenous injection group (0.5 mg / kg) and oral group (20 mg / kg). The rats were fasted within 15 h before the experiment, and were deprived of water within 2 h after administration, and fasted within 4 h. In the oral administration group, about 0.5 mL of blood was collected from the orbital venous plexus before administration and 15 min, 30 min, 45 min, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after administration, and EDTA anticoagulated; in the intravenous administration group, about 0.5 mL of blood was collected from the orbital venous plexus before administration and 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after administration, and EDTA anticoagulated; all blood samples were stored at 4 ° C. Centrifuged at 4000 r / min for 10 min at 4 ° C, and plasma was collected and stored in a refrigerator at -80 ° C. The concentration of compound 2 in plasma samples was detected by LC / MS, and pharmacokinetic parameters were calculated using PKSolver 2.0.

[0371] The formula for calculating oral absolute bioavailability is as follows:

[0372] F(%)=AUC 0-inf口服 *D 静注 / AUC 0-inf静注 *D 口服

[0373] AUC is the area under the drug-time curve, and D is the administered dose.

[0374] The research results show that: Fig.10 As shown in Table 7, compound 2 reached a peak plasma concentration of 30.53 ng / mL 2 hours after oral administration, with an in vivo half-life of 8.03 hours and an area under the drug-time curve of 307.90 ng / mL*h. After intravenous injection of compound 2, the peak plasma concentration was 462.57 ng / mL, the area under the drug-time curve was 2065.12 ng / mL*h, and the half-life was reduced to 6.39 hours compared with oral administration. The calculated absolute oral bioavailability F = 14.90%.

[0375] Example 61: Brain targeting effect experiment of compound 2

[0376] Six SPF male Kunming mice weighing 22-26 g were purchased from the Experimental Animal Center of the School of Pharmacy, Sun Yat-sen University.

[0377] Compound 2 was injected intravenously (2 mg / kg). No food was allowed within 15 hours before the experiment, no water was allowed within 2 hours after administration, and no food was allowed within 4 hours. About 0.5 mL of blood was collected 0.5 hours and 2 hours after administration, and the whole brain was distributed. The serum was anticoagulated with EDTA; the whole brain was stored at -80°C. The concentration of compound 2 in plasma samples was detected by LC / MS, and the drug brain / blood distribution ratio was calculated.

[0378] The research results are shown in Table 8. The brain / blood ratios of compound 2 at the two time points of 0.5h and 2h were 0.45 and 0.48 respectively, indicating that compound 2 can be quickly and evenly distributed in plasma and brain tissue within 0.5h after intravenous injection; this indicates that compound 2 is distributed quickly in the body and has the ability to pass through the blood-brain barrier.

[0379] Example 62: In vitro pharmacokinetic studies of compounds 25, 35, 38, 43, and 45

[0380] [Intermediate solution]: intermediate solution obtained by diluting the sample with methanol; [Stop solution]: internal standard + methanol;

[0381] [Liver microparticle working solution]: liver microparticle stock solution + phosphate buffer (ice bath environment);

[0382] [Incubation system]: methanol + water + phosphate buffer (ice bath environment);

[0383] [Incubation working solution]: Add [intermediate solution] to [incubation system] and mix well (in ice bath environment);

[0384] 0min reaction tube: directly add [incubation working solution] + [liver microparticle working solution]; incubate the remaining [incubation working solution] at 37℃ for 5min, then take the [incubation working solution] and mix it with the [liver microparticle working solution], mix it immediately and keep incubating at 37℃, take it out at 5, 15, 30, 60 minutes and add the stop solution (4 bottles at each time point) and vortex to mix it to quickly stop the reaction. After stopping the reaction, keep the sample in an ice bath until detection; after low-temperature centrifugation at 10000rpm, take the supernatant and detect it with triple quadrupole ultra-performance liquid chromatography tandem mass spectrometry, and calculate the half-life based on the results.

[0385] The research results are shown in Table 2. The half-life of the fluorine-substituted compounds is significantly prolonged, indicating that different side chain substitutions have a significant effect on the metabolism of the compounds.

[0386] Example 63: Experiment on the anti-acute colitis effect of compound 24

[0387] (1) Methods: 90 C57BL / 6J mice were randomly divided into 9 groups according to body weight, with 10 mice in each group, namely A. blank control group, B. model control group, C. low-dose (5 mg / kg) group of compound 24, D. medium-dose (10 mg / kg) group of compound 24, E. high-dose (20 mg / kg) group of compound 24, and F. positive drug control group (sulfasalazine colon-soluble capsule, SASP, 200 mg / kg). After the quarantine period, except for the blank control group, the remaining groups were given 3.5% DSS, and the mice were allowed to drink freely for 7 days to induce an acute ulcerative colitis model. The blank and model control groups were given solvent (2% Tween 80) by gavage, and each drug group was given low, medium, and high concentrations of the corresponding sample by gavage every day. At the same time, the positive drug group was given SASP by gavage every day until the end of the experiment. The activity signs of the mice, weight changes, stool consistency and bleeding were recorded every day. After 7 days, the mice were completely anesthetized and 1 mL of blood was collected from their eye sockets. The mice were then euthanized by cervical dislocation and samples were taken for relevant index detection and observation.

[0388] (2) Data analysis: The experimental data were statistically processed using GraphPad Prism 9.2 biostatistics software: quantitative data were expressed as Mean ± SD and analyzed using analysis of variance combined with Dunnett's multiple comparison method; categorical data were analyzed using the Kruskal-Wallis rank sum test combined with Dunnett's multiple comparison method; P < 0.05 was considered statistically significant when comparing between groups.

[0389] (3) Research results:

[0390] ① As shown in the results of Table 9, compared with the blank control group, the model control, each drug-treated and positive control group mice had bloody stools starting from the second day of modeling. As the number of modeling days increased, the degree of bloody stools gradually increased (P<0.05), indicating that the model was successfully prepared; compared with the model control group, on the fifth day of modeling, the scores of bloody stools in the drug-treated groups and the positive control group showed a downward trend, but no significant difference was observed (P>0.05); on the sixth day of modeling, the scores of bloody stools in the drug-treated groups and the positive control group showed a downward trend, among which the scores of bloody stools in the high-dose group of compound 24 and the positive drug control group were significantly reduced (P<0.05); on the seventh day of modeling, the scores of bloody stools in the drug-treated groups and the positive control group showed a downward trend, among which the scores of bloody stools in the positive drug control group were significantly reduced (P<0.05).

[0391] ② Effect on the disease activity index (DAI) score: As shown in the results of Table 10, compared with the blank control group, the model control, each drug-treated and positive control group mice had thinner stool consistency and bloody stool starting from the 3rd day of modeling. As the number of modeling days increased, the DAI score gradually increased (P<0.05), indicating that the model was successfully prepared; compared with the model control group, on the 4th and 5th days of modeling, the DAI scores of the drug-treated groups and the positive control group showed a downward trend, but no significant difference was found (P>0.05); on the 6th day of modeling, the DAI scores of the drug-treated groups and the positive control group showed a downward trend, among which the DAI scores of the high-dose compound 24 group and the positive drug control group were significantly reduced (P<0.05); on the 7th day of modeling, the DAI scores of the drug-treated groups and the positive control group showed a downward trend, among which the DAI scores of the high-dose compound 24 group and the positive drug control group were significantly reduced (P<0.05).

[0392] ③ Effect on spleen weight and colon length: As shown in Table 11, compared with the blank control group, the spleen weight and spleen coefficient of the model control group were significantly increased (P<0.05), and the colon length was significantly shortened (P<0.05), indicating that the model was successfully prepared; compared with the model control group, the spleen weight and spleen coefficient of each drug-treated group and the positive control group showed a downward trend, but no significant difference was found (P>0.05); compared with the model control group, the colon length of each drug-treated group and the positive control group showed an upward trend, among which the positive drug control group showed a significant increase (P<0.05).

[0393] Under the conditions of this experiment, compound 24 had a certain improvement effect on DSS-induced acute ulcerative colitis in mice.

[0394] Tables 1-11 during the testing process of the present invention are as follows.

[0395] Table 1 Evaluation of biological activities of compounds 1-23

[0396]

[0397]

[0398] Note: Selectivity Index=RIPK3 IC 50 / RIPK1 IC 50 .

[0399] Table 2 Evaluation of biological activities of compounds 24-48

[0400]

[0401]

[0402]

[0403] Note: R 1 Methyl, Selectivity Index = RIPK3 IC 50 / RIPK1 IC 50 .

[0404] Table 3 Neurological function scores of animals in each group

[0405] Grouping Number of animals (N) Longa Rating MNSS score Sham operation group 12 0.00±0.00 0.00±0.00 Model control group 11 1.73±0.79 7.45±3.39 2 Low dose group 10 1.30±0.95 5.20±2.74* 2 Medium dose group 11 1.27±0.47 5.09±1.87* 2 High dose group 11 1.00±0.00** 3.82±0.40*** Positive control group 9 1.00±0.00* 3.78±0.97***

[0406] Note: Compared with the model group, *p<0.05, **p<0.01, ***p<0.001; n=9-12, data are expressed as Mean±SEM.

[0407] Table 4 Percentage of cerebral infarction volume in each group of animals

[0408] Grouping Number of animals (N) Percentage of cerebral infarction volume (%) Sham operation group 12 0.00±0.00 Model control group 11 24.38±5.16 2 Low dose group 10 20.60±6.15 2 Medium dose group 11 18.32±4.07** 2 High dose group 11 16.51±2.66*** Positive control group 9 19.65±4.03

[0409] Note: Compared with the model group, **p<0.01, ***p<0.001; n=9-12, data are expressed as Mean±SEM.

[0410] Table 5 Serum SOD, MDA, TNF-α and IL-1β levels in rats 24 h after MCAO reperfusion

[0411]

[0412]

[0413] Note: Compared with the model group, **p<0.01, ***p<0.001, ****p<0.0001; n=9-12, data are expressed as Mean±SEM.

[0414] Table 6 Effects on the body weight of Kunming mice within 7 days

[0415] Group Body weight before medication (g) 1 day after medication (g) 3 days after medication (g) 5 days after medication (g) 7 days after medication (g) 2 High dose group 26.70±2.40 25.65±2.58 27.05±2.18 27.33±2.07 28.58±2.06 2 Medium dose group 26.85±1.14 27.10±0.57 27.25±1.70 27.35±1.38 28.10±1.53 2 Low dose group 26.73±0.78 26.50±1.46 28.28±1.28 28.45±1.00 29.20±1.30 2. Solvent control group 26.30±0.57 27.85±1.33 28.55±0.99 28.70±1.00 29.35±1.22

[0416] Note: n=4, Data are expressed as Mean ± SEM.

[0417] Table 7 Pharmacokinetic parameters of compound 2 after oral administration and intravenous injection

[0418]

[0419] Table 8 Brain / blood distribution ratio of compound 2

[0420]

[0421] Note: ***The concentration is abnormal and is quite different from the data in the same group. This group of data is not adopted.

[0422] Table 9 Effects on the degree of blood in stool in mice (n=10, )

[0423]

[0424]

[0425] Note: Compared with the blank control group #: P < 0.05, compared with the model control group *: P < 0.05.

[0426] Table 10 Effects on DAI in mice (n=10, )

[0427]

[0428] Note: Compared with the blank control group #: P < 0.05, compared with the model control group *: P < 0.05.

[0429] Table 11 Effects on spleen coefficient and colon length in mice (n=10, )

[0430]

[0431] Note: Compared with the blank control group #: P < 0.05, compared with the model control group *: P < 0.05.

[0432] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.

Claims

1. A RIPK1 kinase target inhibitor, characterized in that The general structural formula of the RIPK1 kinase target inhibitor is shown in Formula (I) or Formula (II): wherein X, Y, and Z are all nitrogen atoms; or, Z is a carbon atom, X and Y are one of a nitrogen atom and a sulfur atom, and the atoms of X and Y are different; or, Z is a carbon atom, and X and Y are both nitrogen atoms; or, Y and Z are carbon atoms, and X is an oxygen, nitrogen, or sulfur atom; R 1 is selected from one of hydrogen, methyl, ethyl, propyl, isopropyl, phenyl, and substituted phenyl; R 2 and R 3 Each is independently selected from an alkyl group having 6 or less carbon atoms or a halogen-substituted product thereof, a cycloalkyl group having 6 or less carbon atoms or a halogen-substituted product thereof, a phenyl group or a halogen-substituted product thereof, a benzyl group, a substituted benzyl group, a methylene group substituted with an aromatic heterocycle or a halogen-substituted product thereof; G in formula (II) is one of CH2, NH, and O.

2. The RIPK1 kinase target inhibitor according to claim 1, characterized in that The structural formula of the RIPK1 kinase target inhibitor is selected from at least one of the following:

3. A method for preparing the RIPK1 kinase target inhibitor according to any one of claims 1 to 2, characterized in that: The steps include: (1) Synthesis of 6-bromoquinazolinecarboxylic acid amide derivatives: 5-bromoisatin is used as a raw material, subjected to alkaline hydrolysis, and then diluted hydrochloric acid is added to adjust the pH to neutral to obtain a sodium salt of the isatin hydrolysis product; the sodium salt of the isatin hydrolysis product is subjected to a three-component reaction with acetaldehyde and ammonium acetate to obtain a sodium salt of quinazolinecarboxylic acid; the sodium salt of quinazolinecarboxylic acid is condensed with an amine compound in the presence of a condensing agent to produce a 6-bromoquinazolinecarboxylic acid amide derivative, which is fragment A; (2) At least one of the following (2a) to (2d): (2a) Preparation of Fragment B: Using a 7- or 6-brominated 2-amino-benzo-pentacyclic compound as a starting material, adding an acyl chloride or a carboxylic acid, and in the presence of an acid binding agent or a condensing agent, preparing a 7- or 6-brominated benzo-pentacyclic-2-aminoacylated derivative, which is Fragment B; (2b) Preparation of Fragment C: Using a 2-amino-benzo-pentacyclic compound brominated at the 6-position as a raw material, a carboxylic acid is added, and in the presence of a condensing agent, a benzo-pentacyclic 2-aminoacylated derivative substituted with a bromomethyl group at the 6-position is prepared, which is Fragment C; (2c) preparing fragment D, using a 2-amino-benzo five-membered heterocycle substituted with a nitro group at the 6-position as a raw material, adding a carboxylic acid, in the presence of a condensing agent, to prepare a benzo five-membered heterocycle substituted with a nitro group-2-aminoacylated derivative, and adding a nitro reducing agent to prepare the corresponding benzo five-membered heterocycle substituted with an amino group-2-aminoacylated derivative, which is fragment D; (2d) preparing fragment E, using a 2-amino-benzo five-membered heterocycle substituted with a hydroxyl group at the 6-position as a starting material, adding a phenolic hydroxyl protecting agent to protect the hydroxyl group, adding a carboxylic acid, and preparing a benzo five-membered heterocycle-2-amino acylated derivative protected with a silyl ether at the 6-position in the presence of a condensing agent, and adding a deprotecting agent to prepare the corresponding benzo five-membered heterocycle-2-amino acylated derivative substituted with a phenolic hydroxyl group at the 6-position, which is fragment E; (3) At least one of the following (3a) to (3c): (3a) reacting the fragment B prepared in step (2) with pinacol diboron in the presence of an organic base A and a palladium catalyst to prepare an aryl borate of the fragment B, which is then reacted with the fragment A prepared in step (1) in the presence of an organic base B, a palladium catalyst, and a phosphine ligand to obtain a RIPK1 kinase target inhibitor represented by formula (I); the organic base A is potassium acetate, and the organic base B is at least one of Cs2CO3 and K3PO4; (3b) reacting the fragment A prepared in step (1) with pinacol diboron in the presence of an organic base A and a palladium catalyst to prepare the corresponding aryl borate, and then reacting it with the fragment C prepared in step (2) in the presence of an organic base B, a palladium catalyst, and a phosphine ligand to obtain a RIPK1 kinase target inhibitor represented by formula (II); (3c) reacting the fragment D or fragment E prepared in step (2) with the fragment A prepared in step (1) in the presence of an organic base B, a palladium catalyst, and a phosphine ligand to obtain a RIPK1 kinase target inhibitor represented by formula (II).

4. The method for preparing the RIPK1 kinase target inhibitor according to claim 3, wherein: In the step (1): The base is at least one of sodium hydroxide and potassium hydroxide; And / or, the condensing agent is one of mixture A and mixture B, wherein mixture A is a mixture of N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate and N-methylimidazole, and the molar ratio of the mixture of N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate and N-methylimidazole is 1:2-3; and mixture B is a mixture of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 1-hydroxybenzotriazole, and triethylamine, and the molar ratio of these is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride:1-hydroxybenzotriazole:triethylamine=1:1:1; and / or, the amine compound is (R)-1-phenylethylamine, (S)-1-phenylethylamine, (S)-1-phenylpropylamine, (S)-2-methyl-1-phenylpropylamine, (S)-1-cyclohexylethylamine, (S)-1-(4-methylphenyl)ethylamine, (S)-1-(4-methoxyphenyl)ethylamine, (S)-1-(4-nitrophenyl)ethylamine, (S)-1-(pyridin-2-yl)ethylamine, (S)- At least one of 1-(pyridin-3-yl)ethylamine, (S)-1-(pyridin-4-yl)ethylamine, (S)-1-(2-fluorophenyl)ethylamine, (S)-1-(3-fluorophenyl)ethylamine, (S)-1-(4-fluorophenyl)ethylamine, aniline, benzylamine, phenethylamine, tetrahydro-2H-pyran-4-amine, 4-trifluoromethylbenzylamine, 3-trifluoromethylbenzylamine, 4-trifluoromethoxybenzylamine, and 3-trifluoromethoxybenzylamine; And / or, the molar ratio of 5-bromoisidine and base is 5-bromoisidine:base=1:1-3; And / or, the molar ratio of the sodium salt of isatin hydrolysate, acetaldehyde, and ammonium acetate is: sodium salt of isatin hydrolysate: acetaldehyde: ammonium acetate = 1:1-3:2-5; And / or, the molar ratio of the quinazoline carboxylic acid sodium salt, the amine compound, and the condensing agent is 1:1:2-3;.

5. The method for preparing the RIPK1 kinase target inhibitor according to claim 3, wherein: In the steps (2a) to (2d): The acyl chloride is at least one of n-butyryl chloride, isobutyryl chloride, n-valeryl chloride, pivaloyl chloride, cyclopropylcarbonyl chloride, cyclobutylcarbonyl chloride, cyclopentylcarbonyl chloride, cyclohexylcarbonyl chloride, and 3-oxocyclobutylcarbonyl chloride; and / or the carboxylic acid is at least one of acetic acid, propionic acid, n-butyric acid, isobutyric acid, n-valeric acid, isovaleric acid, pivalic acid, cyclopropanecarboxylic acid, cyclobutanecarboxylic acid, cyclopentanecarboxylic acid, cyclohexanecarboxylic acid, and benzoic acid; And / or, the acid binding agent is at least one of triethylamine and N,N-diisopropylethylamine; And / or, the condensing agent is one of mixture A and mixture B, wherein mixture A is a mixture of N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate and N-methylimidazole, and the molar ratio of the mixture of N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate and N-methylimidazole is 1:2-3; and mixture B is a mixture of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 1-hydroxybenzotriazole, and triethylamine, and the molar ratio of these is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride:1-hydroxybenzotriazole:triethylamine=1:1:1; and / or, the nitro reducing agent is stannous chloride; And / or, the phenolic hydroxyl protecting agent is a mixture of tert-butyldimethylchlorosilane and imidazole, and the molar ratio of tert-butyldimethylchlorosilane to imidazole is 1:1-2; And / or, the deprotection reagent is tetrabutylammonium fluoride.

6. The method for preparing the RIPK1 kinase target inhibitor according to claim 3, wherein: In the step (2a), the molar ratio of the 2-amino-benzo five-membered heterocyclic compound, the acyl chloride, and the acid-binding agent is 2-amino-benzo five-membered heterocyclic compound: acyl chloride: acid-binding agent = 1:1-2:1-3; And / or, in steps (2a), (2b), (2c), and (2d), the molar ratio of the 2-amino-benzo five-membered heterocyclic compound, carboxylic acid, and condensing agent is 2-amino-benzo five-membered heterocyclic compound: carboxylic acid: condensing agent = 1:1:1.1-1.5; And / or, in the step (2c), the molar ratio of the benzo-5-membered heterocyclic-2-aminoacylated derivative substituted with a nitro group at the 6-position and the nitro reducing agent is 1:3-6. And / or, in the step (2d), the molar ratio of the 2-amino-benzo five-membered heterocycle substituted with the 6-hydroxyl group to the phenolic hydroxyl protecting agent is 1:1-3; And / or, in the step (2d), the molar ratio of the 6-silyl ether protected benzo five-membered heterocycle-2-aminoacylated derivative and the deprotection reagent is 6-silyl ether protected benzo five-membered heterocycle-2-aminoacylated derivative: deprotection reagent = 1:2-5.

7. The method for preparing the RIPK1 kinase target inhibitor according to claim 3, wherein: In the steps (3a), (3b) and (3c), the palladium catalyst is at least one of Pd(dppf)Cl2 and Pd(dppf)Cl2·CH2Cl2, and the phosphine ligand is PCy3; And / or, in the step (3a), the molar ratio of the fragment B prepared in the step (2), the pinacol diborate, the palladium catalyst, and the organic base A is: fragment B: pinacol diborate: palladium catalyst: organic base A = 1: 2-3: 0.01-0.3: 2-5; And / or, in step (3a), the molar ratio of the aryl borate of fragment B, fragment A, palladium catalyst, phosphine ligand, and organic base B is aryl borate of fragment B: fragment A: palladium catalyst: phosphine ligand: organic base B = 1:1:0.01-0.2:0.01-0.2:2-5; And / or, in the step (3b), the molar ratio of the fragment A prepared in step (1), the pinacol diborate, the palladium catalyst, and the organic base A is: fragment A:pinacol diborate:palladium catalyst:organic base A=1:2-3:0.01-0.3:2-5; And / or, in step (3b), the molar ratio of the aryl borate of fragment A, fragment C, palladium catalyst, phosphine ligand, and organic base B is aryl borate of fragment A: fragment C: palladium catalyst: phosphine ligand: organic base B = 1:1:0.01-0.2:0.01-0.2:2-5; And / or, in the step (3c), the molar ratio of fragment A prepared in step (1), fragment D or fragment E prepared in step (2), palladium catalyst, phosphine ligand, and organic base B is fragment A: fragment D / fragment E: palladium catalyst: phosphine ligand: organic base B = 1:1:0.01-0.2:0.01-0.2:2-5.

8. A pharmaceutical composition, characterized in that The pharmaceutical composition contains the RIPK1 kinase target inhibitor according to any one of claims 1 to 2.

9. Use of the RIPK1 kinase target inhibitor according to any one of claims 1 to 2 or the pharmaceutical composition according to claim 8 in the preparation of a medicament for treating neurodegenerative diseases, inflammation, autoimmune diseases, and sepsis.

10. A drug, characterized in that The drug contains the RIPK1 kinase target inhibitor according to any one of claims 1 to 2 or the pharmaceutical composition according to claim 8.

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