Cathepsin K inhibitor, its manufacturing method and application

Abstract

The structure is represented by formula (0) and / or formula (II): As shown in JPEG2025503082000036.jpg42160, the present invention relates to a cathepsin K inhibitor in which a pyrimidine cyano group or a ketone group is an important electrophilic group and which exhibits a good inhibitory effect against cathepsin K. Also provided are a production method and application thereof. The provided cathepsin K inhibitor has high inhibitory effect and selectivity, and is expected to be applied to the treatment of diseases including thyroid disease, cardiovascular disease, bone disease, and gum disease.

Description

[Technical field]

[0001] The present invention relates to the field of medicinal chemistry, in particular to compounds that inhibit the activity of cathepsin K, their preparation and their use in the manufacture of medicaments for the treatment of diseases associated with or characterized by the activity of cathepsin K, such as osteoporosis. [Background technology]

[0002] Osteoporosis, abbreviated as osteoporosis, is a metabolic disorder disease in which the total body bone mass is reduced, bone density is reduced, and the patient's risk of fracture is increased. With the progress of population aging, the health problems related to the disease are becoming more and more serious, and the probability of fracture in osteoporosis patients is significantly increased, which places a heavy burden on society and families and has a serious negative impact on people's lives. The mechanism of osteoporosis is due to the imbalance between osteoclast-mediated bone resorption and osteoblast-mediated bone formation. Currently, the commonly used osteoporosis treatment drugs on the market include bone resorption inhibitors such as bisphosphonates and bone formation promoters, but these two drugs have certain defects. Long-term use of bone resorption inhibitors will have a negative effect on the differentiation and proliferation of bone cells, and will further cause the occurrence of "low bone transformation state", while bone formation promoters will increase the probability of patients suffering from osteosarcoma.

[0003] Since all drugs currently available on the market have certain deficiencies, the current research direction is to develop new osteoporosis treatment drugs that have a therapeutic effect on osteoporosis without adversely affecting the proliferation and differentiation of osteoblasts and osteoclasts, and can avoid the occurrence of low bone turnover state.

[0004] Cathepsin K (Cat K) belongs to the papain family of cysteine ​​proteases, and is abundant in osteoclasts. It is currently a new target for osteoporosis treatment. Human bone matrix is ​​composed of 25% water, 25% organic matrix, and 50% mineral matrix, of which 90% of the organic component is type I collagen. The degradation of type I collagen is an important process of osteoclast-mediated bone resorption, and Cat K plays a dominant role in the degradation of type I collagen. Cat K inhibitors can block the degradation of matrix collagen, thereby inhibiting the resorption of mature osteoclasts. At the same time, Cat K inhibitors can maintain the survival number of osteoclasts, which ensures that the binding signal between osteoclasts and osteoblasts is fully maintained, thereby avoiding the occurrence of "low bone conversion state". Cat K also plays an important role in the pathogenic process of thyroid disease, cardiovascular disease, and gingival disease. The diseases characterized by abnormal expression or activation of Cat K include thyroid disease, cardiovascular disease, bone disease and gingival disease, specifically hyperthyroidism, atherosclerosis, myocardial hypertrophy, heart failure, osteoporosis, osteoarthritis, rheumatoid arthritis, gingivitis and periodontitis. In recent years, there has been an increasing number of studies on Cat K, and it can cause an increase in Cat K secreted by endothelial cells in pathological conditions such as coronary artery sclerosis. Cat K is closely related to the development of myocardial hypertrophy and heart failure. Garg et al. (Garg G, Pradeep AR, Thorat MK, et al. Effect of nonsurgical periodontal therapy on crevicular fluid levels of Cathepsin K in periodontitis. Arch Oral Biol, 54: 1046-1051) demonstrated that the level of Cat K in gingival crevicular fluid before and after baseline treatment in patients with periodontitis decreased after baseline treatment, along with a decrease in clinical gingival index, periodontal probing depth and attachment loss in patients with periodontitis. Cat K should be further studied and paid attention to in periodontitis treatment as a marker of bone resorption in periodontitis.

[0005] At present, the activity of Cat K inhibitor and various inhibitors reported in clinical research stage are all greatly improved.However, there is still no drug on the market for this target, the main reason is that the compound has low selectivity and has high inhibitory activity against other subtypes, such as B type and S type, which leads to the occurrence of drug side effects.Therefore, aiming at reducing side effects, the development and research of new Cat K inhibitors that are efficient and selective is the main research direction at present.

[0006] In view of the fact that Cat B and Cat K belong to the same cysteine ​​protease, Cat L is an acidophilic protease, and Cat L is difficult to obtain, the cyano-substituted pyrimidine compound designed and synthesized in the present invention has good selectivity when its activity against K, B and S is measured, and is expected to be applied to the treatment of diseases related to or characterized by Cat K activity. Summary of the Invention

[0007] The present invention provides novel ketone compounds and / or nitrile compounds acting on Cat K and pharma- ceutically acceptable salts thereof, as well as methods for producing the same and applications thereof.

[0008] The present invention relates to a compound having the general formula: [ka] [In the formula 0, X is C or N, R1 is selected from H, a substituted or unsubstituted C1-10 alkyl group, and a substituted or unsubstituted C3-C10 cycloalkyl group, wherein the substituent is selected from a halogen, an amino group, a cyano group, a hydroxyl group, an aldehyde group, a carboxyl group, and a sulfone group; R * is selected from halogen or a C1-6 alkyl group; Y is a cyclic group, which is located at any position of the aromatic ring to which it is linked, and which is bonded to the aromatic ring by one or two carbon atoms, and which may be a C3-10 cycloalkyl group, a C6-12 aromatic ring, or a C5-12 heterocycle; the C6-12 aromatic ring includes a C6-12 aromatic ring and a C6-12 aromatic heterocycle, the C6-12 aromatic heterocycle contains at least one heteroatom, the C5-12 heterocycle is a saturated or unsaturated heterocycle, the heterocycle contains 1-3 heteroatoms, the heteroatom may be O, N or S; e is 0 or 1, and when e is 0, R5 is directly bonded to any position of the aromatic ring; R5 is selected from H, halogen, an amino group, a cyano group, a C1-10 alkyl group, a C1-10 alkoxy group, a C3-10 cycloalkyl group, a substituted or unsubstituted C3-10 heterocycloalkyl group, -S(O)2R2, -C(O)R2, -NR3R4, -C(O)NHR7, -SR6, and -OR6; The substituents of the C3-10 heterocycloalkyl group are selected from a hydroxy group and a C1-10 alkyl group; The C1-10 alkyl group may be further substituted with a hydroxy group, -C(O)R2; R2 is selected from H, an amino group, a halogen atom, a substituted or unsubstituted C1-6 alkyl group, a C1-6 alkoxy group, a C3-8 cycloalkyl group, or a C3-8 heterocycloalkyl group, and the substituent of the C1-6 alkyl group, C1-6 alkoxy group, C3-8 cycloalkyl group, or C3-8 heterocycloalkyl group is a C1-6 alkyl group; R3 and R4 are each independently selected from H and -C(O)R8, or R3 and R4 together with the N to which they are attached form a 4-8 membered ring containing at least one N, and R8 is selected from a C1-6 alkyl group substituted with a piperazinyl group or a methylpiperazine group; R6 is selected from C1-6 alkyl groups; R7 is selected from a C1-6 alkyl group, which may be further substituted with -C(O)R9, and R9 is selected from a piperazinyl group or a methylpiperazine group; said C3-10 heterocycloalkyl group or C3-8 heterocycloalkyl group contains 1-3 heteroatoms, said heteroatoms may be O, N or S; The halogen is for mono- or polysubstitution and is selected from F, Cl, Br, I. The present invention provides a nitrile compound having a structure represented by the following formula:

[0009] When t is 0 and R* is H, the compound of formula 0 has formula I: [ka] In formula I, X is C or N, R1 is selected from H, a substituted or unsubstituted C1-10 alkyl group, and a substituted or unsubstituted C3-C10 cycloalkyl group, wherein the substituent is selected from a halogen, an amino group, a cyano group, a hydroxyl group, an aldehyde group, a carboxyl group, and a sulfone group; Y is a cyclic group, which is located at any position of the aromatic ring to which it is linked and may be a C3-10 cycloalkyl group, a C6-12 aromatic ring, or a C5-12 heterocycle; the C6-12 aromatic ring includes a C6-12 aromatic ring and a C6-12 aromatic heterocycle, the C6-12 aromatic heterocycle contains at least one heteroatom, the C5-12 heterocycle is a saturated or unsaturated heterocycle, the heterocycle contains 1-3 heteroatoms, the heteroatom may be O, N or S; R5 is selected from H, a halogen, an amino group, a cyano group, a C1-10 alkyl group, a C1-10 alkoxy group, a C3-10 cycloalkyl group, -S(O)2R2, -C(O)R2, -NR3R4, -SR6, and -OR6, wherein R2 is selected from H, an amino group, a halogen, a C1-6 alkyl group, a C1-6 alkoxy group, and a C3-8 cycloalkyl group, wherein R3 and R4 together with the N to which they are attached form a 4-8 membered ring containing at least one N, and R6 is selected from a C1-6 alkyl group; The halogen is for mono- or polysubstitution and is selected from F, Cl, Br, I. As expressed by:

[0010] Further, X is C; R1 is selected from a C1-6 alkyl group, a C4-C8 cycloalkyl group; Y is a cyclic group, which is located at the para position of the aromatic ring and may be a C4-8 cycloalkyl group, a C6-10 aromatic ring, or a C5-10 heterocycle; the C6-10 aromatic ring includes a C6-10 aromatic ring and a C6-10 aromatic heterocycle, the C6-10 aromatic heterocycle containing at least one heteroatom, the C5-10 heterocycle is a saturated or unsaturated heterocycle, the heterocycle containing 1-3 heteroatoms, the heteroatom may be O, N or S; R5 is selected from H, a halogen, an amino group, a cyano group, a C1-6 alkyl group, a C1-6 alkoxy group, a C3-8 cycloalkyl group, -S(O)2R2, -C(O)R2, -NR3R4, -SR6, and -OR6, wherein R2 is selected from H, an amino group, a halogen, a C1-6 alkyl group, a C1-6 alkoxy group, and a C3-8 cycloalkyl group, wherein R3 and R4 together with the N to which they are attached form a 5-8 membered ring containing at least one N, and wherein R6 is selected from a C1-6 alkyl group.

[0011] The halogen is for mono- or polysubstitution and is selected from F, Cl, Br, I.

[0012] Further, X is C; R1 is selected from a C1-6 alkyl group and a C5-8 cycloalkyl group; Y is a cyclic group, which is located at the para position of the aromatic ring and may be a C5-8 cycloalkyl group, a C6-8 aromatic ring, or a C5-8 heterocycle; the C6-8 aromatic ring includes a C6-8 aromatic ring and a C6-8 aromatic heterocycle, the C6-8 aromatic heterocycle containing at least one heteroatom, the C5-8 heterocycle is a saturated or unsaturated heterocycle, the heterocycle containing 1-3 heteroatoms, the heteroatom may be O, N or S; R5 is selected from H, a halogen, an amino group, a cyano group, a C1-3 alkyl group, a C1-3 alkoxy group, -S(O)2R2, -C(O)R2, -NR3R4, -SR6, and -OR6, wherein R2 is selected from an amino group, a halogen, and a C1-3 alkyl group, wherein R3 and R4 together with the N to which they are attached form a 5-8 membered ring containing at least one N, and wherein R6 is selected from a C1-3 alkyl group.

[0013] The halogen is for mono- or polysubstitution and is selected from F, Cl, Br, I.

[0014] In some embodiments, R1 is selected from a neopentyl group and a cyclohexyl group.

[0015] In some embodiments, Y is [ka] , a phenyl group, a pyridyl group, a thienyl group, and a thiazolyl group.

[0016] In some preferred embodiments, Y is [ka] , a phenyl group, and a thiazolyl group.

[0017] In some embodiments, R5 is selected from H, F, Cl, a cyano group, a methyl group, a methylthio group, a methoxy group, a methylsulfonyl group, a methylcarbamoyl group, and a methylpiperazine group.

[0018] In some preferred embodiments, R5 is selected from a methyl group and a methylpiperazine group.

[0019] The present invention relates to a compound having the structure shown in the general formula: [ka] [In the formula, R1 and R2 are each independently selected from H, a halogen, a cyano group, an amino group, a substituted or unsubstituted C1-6 alkyl group, and a substituted or unsubstituted C1-6 alkoxy group, and the substituted C1-6 alkyl group or the substituted C1-6 alkoxy group is further substituted with at least one halogen or hydroxy group; t is a chemical bond, [ka] At least one selected from the following may be used. The present invention provides a keto-based cathepsin K inhibitor comprising the compound:

[0020] Further, R1 and R2 are each independently selected from H, a halogen, a C1-3 alkyl group, and a C1-3 alkoxy group, and the C1-3 alkyl group or the C1-3 alkoxy group is further substituted with at least one halogen.

[0021] Further, R1 and R2 are each independently selected from H, halogen, a C1-3 alkyl group, and a C1-3 alkoxy group, and the C1-3 alkyl group or the C1-3 alkoxy group is further substituted with at least one F.

[0022] The C1-3 alkyl group is a methyl group, an ethyl group, a propyl group, or an isopropyl group.

[0023] The C1-3 alkoxy group is a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, a monofluoromethoxy group, a difluoromethoxy group, or a trifluoromethoxy group.

[0024] The C1-3 alkoxy group is a methoxy group or a trifluoromethyl group.

[0025] The C1-3 alkoxy group is a trifluoromethyl group.

[0026] In some embodiments, R1 and R2 are each independently selected from H, F, Cl, Br, a methoxy group, and a trifluoromethoxy group.

[0027] In some preferred embodiments, R1, R2 are each independently selected from H and F.

[0028] The halogen is selected from F, Cl, Br, I.

[0029] The compound provided by the present invention is 1) 2-[(2,2-dimethylpropyl){[4-(4-methylpiperazin-1-yl)phenyl]methyl}amino]pyrimidine-4-carbonitrile, 2) 2-[(2,2-dimethylpropyl)[(4-phenylphenyl)methyl]amino]pyrimidine-4-carbonitrile, 3) 2-[(2,2-dimethylpropyl)({4-[4-(methylthio)phenyl]phenyl}methyl)amino]pyrimidine-4-carbonitrile, 4) 2-[(2,2-dimethylpropyl){[4-(4-fluorophenyl)phenyl]methyl}amino]pyrimidine-4-carbonitrile, 5) 2-[(2,2-dimethylpropyl){[4-(4-methoxyphenyl)phenyl]methyl}amino]pyrimidine-4-carbonitrile, 6) 2-[(2,2-dimethylpropyl)({4-[4-(methylsulfonyl)phenyl]phenyl}methyl)amino]pyrimidine-4-carbonitrile, 7) 2-({[4-(5-cyanothiophen-2-yl)phenyl]methyl}(2,2-dimethylpropyl)amino)pyrimidine-4-carbonitrile, 8) 2-({[4-(6-chloropyridin-3-yl)phenyl]methyl}(2,2-dimethylpropyl)amino)pyrimidine-4-carbonitrile, 9) 2-({[4-(3,4-dichlorophenyl)phenyl]methyl}(2,2-dimethylpropyl)amino)pyrimidine-4-carbonitrile, 10) 2-[(2,2-dimethylpropyl){[4-(5-methylthiophen-2-yl)phenyl]methyl}amino]pyrimidine-4-carbonitrile, 11) 4-(4-{[(4-cyanopyrimidin-2-yl)(2,2-dimethylpropyl)amino]methyl}phenyl)methylbenzoate, 12) 2-({[4-(4-chloro-3-fluorophenyl)phenyl]methyl}(2,2-dimethylpropyl)amino)pyrimidine-4-carbonitrile, 13) 2-[(2,2-dimethylpropyl)({4-[4-(4-methylpiperazin-1-yl)phenyl]phenyl}methyl)amino]pyrimidine-4-carbonitrile, 14) 2-[(cyclohexylmethyl)[(4-phenylphenyl)methyl]amino]pyrimidine-4-carbonitrile, 15) 2-[(cyclohexylmethyl)({4-[4-(4-methylpiperazin-1-yl)phenyl]phenyl}methyl)amino]pyrimidine-4-carbonitrile, 16) 2-[(cyclohexylmethyl)({4-[4-(methylsulfonyl)phenyl]phenyl}methyl)amino]pyrimidine-4-carbonitrile, 17) 2-(neopentyl(4-(thiazol-4-yl)benzyl)amino)pyrimidine-4-carbonitrile, 18) 2-((4-(2-(4-methylpiperazin-1-yl)thiazol-4-yl)benzyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 19) (1R,2R)-2-(8-fluoro-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-2-carbonyl)-N-(4-oxotetrahydrofuran-3-yl)cyclohexane-1-formamide; 20) (1R,2R)-2-(2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 21) (1R,2R)-2-(8-fluoro-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-2-carbonyl)-N-((S)-4-oxotetrahydrofuran-3-yl)cyclohexane-1-formamide; 22) (1R,2R)-2-(6,8-difluoro-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-2-carbonyl)-N-((S)-4-oxotetrahydrofuran-3-yl)cyclohexane-1-formamide; 23) (1R,2R)-2-(8-chloro-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-((S)-4-oxotetrahydrofuran-3-yl)cyclohexane-1-formamide; 24) (1R,2R)-2-(8-bromo-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-((S)-4-oxotetrahydrofuran-3-yl)cyclohexane-1-formamide; 25) (1R,2R)-2-(6-(trifluoromethoxy)-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 26) (1R,2R)-2-(6-(trifluoromethoxy)-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(S)-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 27) (1R,2R)-2-(6-fluoro-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(S)-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 28) (1R,2R)-2-(6-fluoro-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(S)-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 29) (1R,2R)-2-(6-methoxy-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(S)-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 30) (1R,2R)-2-(6-methoxy-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 31) (1R,2R)-2-(8-fluoro-6-methoxy-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(S)-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 32) (1R,2R)-2-(8-fluoro-6-methoxy-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 33) 2-((4-chlorobenzyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 34) 2-((4-bromobenzyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 35) 2-((naphthalen-2-ylmethyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 36) 2-(neopentyl(quinolin-2-ylmethyl)amino)pyrimidine-4-carbonitrile, 37) 2-(((4'-(4-(2-hydroxyethyl)piperazin-1-yl)-[1,1'-biphenyl]-4-yl)methyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 38) 2-(((4'-(4-hydroxypiperidin-1-yl)-[1,1'-biphenyl]-4-yl)methyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 39) 2-(((2-methyl-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)methyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 40) 2-(((2-chloro-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)methyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 41) 2-(neopentyl(4-(4-(2-oxopropyl)piperazin-1-yl)benzyl)amino)pyrimidine-4-carbonitrile, 42) 2-(isobutyl((4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)methyl)amino)pyrimidine-4-carbonitrile, 43) N-(4-((4-cyanopyrimidin-2-yl)(neopentyl)amino)methyl)phenyl)-2-(4-methylpiperazin-1-yl)acetamide, 44) 2-((4-(4-methylpiperazine-1-carbonyl)benzyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 45) Contains the specific structure 4-(((4-cyanopyrimidin-2-yl)(neopentyl)amino)methyl)-N-(2-(4-methylpiperazin-1-yl)-2-oxoethyl)benzamide.

[0030] Another object of the present invention is to provide a process for the preparation of the above compounds and pharma- ceutically acceptable salts thereof, the process for the preparation of formula I comprising the steps of: [ka] This includes the steps: where R1, R5, X, and Y are as defined above; L is a halogen selected from F, Cl, and Br; Step 1): Reacting intermediate T3 and compound T4 to produce compound T5; Reaction conditions: carried out in alkaline medium, The base is an inorganic base and is at least one selected from cesium carbonate, sodium carbonate, potassium carbonate, sodium hydrogen carbonate, potassium hydrogen carbonate, aqueous ammonia, sodium hydride, sodium hydroxide, potassium hydroxide, and calcium hydroxide; The solvent is at least one selected from water, methanol, ethanol, glycerin, propylene glycol, formamide, acetonitrile, n-butanol, dioxane, dichloromethane, chloroform, acetone, dimethylsulfoxide, dimethylformamide, ethyl acetate, and tetrahydrofuran; Step 2): Reacting compound T5 with compound T6 to produce a compound of formula I; Reaction conditions: Heat reaction in an alkaline solvent, The base and the solvent are as defined in step 1), The heating temperature is 40-100°C.

[0031] The process for preparing the compound of formula II comprises: [ka] This includes the steps: Step 1) reacting compound P1 with (3aR,7aS)-hexahydroisobenzofuran-1,3-dione to produce compound P2; Reaction conditions: the reaction is carried out in a solvent, and the solvent is a polar solvent, which is at least one selected from water, methanol, ethanol, glycerin, propylene glycol, formamide, acetonitrile, n-butanol, dioxane, dichloromethane, acetone, dimethylsulfoxide, dimethylformamide, ethyl acetate, and tetrahydrofuran; Step 2) reacting compound P2 with 4-aminotetrahydrofuran-3-ol to produce compound P3; Reaction conditions: carried out in a polar solvent containing a base, the base being at least one selected from pyridine, 2-methylpyridine, 2,6-lutidine, 4-dimethylaminopyridine, triethylamine, diethylamine, N,N-diisopropylethylamine, dimethylisopropylamine, piperidine, 1-methylpiperidine, 1-methylpyrrolidine, the polar solvent being as defined according to step 1; Step 3) Oxidizing compound P3 to produce a compound of formula I; Reaction conditions: under oxidizing agent conditions, the oxidizing agent being an inorganic oxidizing agent or an organic oxidizing agent or a mixed oxidizing agent; The inorganic oxidizing agent is at least one selected from sodium dichromate, chromic acid, manganese dioxide, cerium ammonium nitrate, potassium permanganate, potassium perferrate, bromine alone, iodine alone, sodium hypochlorite, sodium chlorite, sodium bromate, sodium periodate, and diiodine pentoxide; the organic oxidizing agent is at least one selected from 2-iodobenzoic acid, tetramethylpiperidine oxide, benzoyl peroxide, and cyclohexanone peroxide; and the mixed oxidizing agent is at least one selected from chlorochromate pyridine salt, chromate pyridine anhydride complex, pyridine dichromate, and sulfur trioxide pyridine complex; Further, the oxidizing agent is preferably a pyridinium chlorochromate salt, a pyridinium chromate anhydride complex, or a pyridinium sulfur trioxide complex.

[0032] The present invention further provides compositions containing said Cathepsin K inhibitors, comprising a therapeutically effective amount of one or more of said compounds or pharma- ceutically acceptable salts thereof.

[0033] The present invention further provides the application of said cathepsin K inhibitor in the manufacture of drugs for treating diseases targeting cathepsin K.

[0034] Furthermore, the diseases targeting cathepsin K include thyroid diseases, cardiovascular diseases, bone diseases, gum diseases and tumors.

[0035] Additionally, the thyroid disease includes hyperthyroidism.

[0036] Furthermore, said cardiovascular diseases include atherosclerosis, myocardial hypertrophy, and heart failure.

[0037] Further, the bone disease includes osteoporosis, osteoarthritis, and rheumatoid arthritis.

[0038] Further, the gum disease includes gingivitis and periodontitis.

[0039] Furthermore, said tumors include tumor invasion and tumor metastasis.

[0040] Furthermore, the disease targeted by cathepsin K is osteoporosis.

[0041] Compared with the prior art, the advantages and technical effects of the present invention are as follows:

[0042] The compounds of the present invention have high inhibitory rates against Cat K enzyme and IC 50 The compound has good inhibitory activity against Cat K enzyme, with an IC value of 10 μM and an inhibition rate of more than 90% against Cat K enzyme. 50 is less than 50 nM.

[0043] The compounds of the present invention have high selectivity. At the levels of 1 μM and 10 μM, the inhibition rates against Cat B enzyme and Cat S enzyme were both <50%, and the IC 50 All values ​​were >10 μM, indicating that the compounds of the present invention have high selectivity for the Cat K enzyme.

[0044] The compounds of the present invention have high drug formability.

[0045] The cathepsin K inhibitors can be used to treat diseases characterized by abnormal expression or activation of cathepsin K, including thyroid disease, cardiovascular disease, bone disease and oral disease, specifically hyperthyroidism, atherosclerosis, myocardial hypertrophy, heart failure, osteoporosis, osteoarthritis, rheumatoid arthritis, gingivitis and periodontitis. The compounds of the present invention can inhibit the generation of osteoclasts and can be used to treat osteoporosis.

[0046] Terms of the invention: 1. Abbreviations and Definitions DMF: N,N-dimethylformamide DMSO: Dimethyl sulfoxide DMAP: 4-dimethylaminopyridine DCM: dichloromethane HATU: 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate DIPEA: N,N-diisopropylethylamine 2. Other terms The term "C1-10 alkyl group" refers to a straight or branched chain hydrocarbon group containing 1-10 carbon atoms and derived by removing one hydrogen atom on a single carbon atom of a parent alkyl group, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, neopentyl, n-hexyl, etc. The term "C1-6 alkyl" refers to any straight or branched chain group containing 1-6 carbon atoms.

[0047] The term "C1-10 alkoxy group" includes an -O-C1-10 alkyl group, meaning that the C1-10 alkyl group is attached to an oxygen atom.

[0048] The term "C1-6 alkoxy group" includes -O-C1-10 alkyl.

[0049] The term "C3-10 cycloalkyl group" refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon group composed solely of carbon and hydrogen atoms, which may include fused, bridged, or spiro ring systems, having 3-10 carbon atoms, and which may be saturated or unsaturated and connected to the remainder of the molecule by a single bond through any suitable carbon atom. Unless otherwise specified herein, the carbon atoms in a cycloalkyl group may be optionally oxidized. Illustrative examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like.

[0050] The term "C6-12 membered aromatic ring" refers to a planar ring system having conjugation, where the bonds between the atoms are not alternating with discrete single and double bonds, but are covered by a delocalized π-electron cloud, and has 6-12 atoms which may be carbon, nitrogen, sulfur, or oxygen atoms. Unless otherwise specified in this specification, the aromatic ring may be a monocyclic, bicyclic, tricyclic or higher ring system.

[0051] The terms "C1-10", "C1-C10" and the like refer to groups containing 1-10 carbon atoms.

[0052] The term "C6-12 aromatic heterocycle" refers to a monocyclic or bicyclic aromatic ring containing 6-12 ring atoms, where 1, 2, 3, 4 ring atoms are selected from nitrogen, sulfur, or oxygen, and the nitrogen or sulfur in the ring may be oxidized. For the purposes of the present invention, the aromatic heterocycle is preferably a stable 5-10 membered aromatic group containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur, more preferably a stable 5-8 membered aromatic group containing 1-2 heteroatoms selected from nitrogen, oxygen, and sulfur. Examples of aromatic groups include, but are not limited to, thienyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, oxadiazolyl, isoxazolyl, pyridyl, pyrimidine, pyrazinyl, and pyridazinyl groups.

[0053] The term "C5-12 heterocycle" refers to a saturated, unsaturated or partially saturated monocyclic or bicyclic ring containing 5-12 ring atoms, in which one, two or three ring atoms are selected from nitrogen, sulfur or oxygen, said rings may be linked by carbon or nitrogen, -CH2- groups in the rings may be replaced by -C(O) groups, nitrogen or sulfur atoms in the rings may be oxidized to form N-oxides or S-oxides, and -NH- in the rings may be replaced by acetyl, formyl, methyl or methanesulfonyl groups.

[0054] Unless otherwise defined, all scientific terms herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0055] It should be understood that the above brief description and the following detailed description are for the purpose of understanding only and are not intended to limit the subject matter of the present invention. In addition, the use of the term "comprises" and other forms thereof, such as "comprises", "contains" and "containing", is not limiting.

[0056] The section titles used herein are for organizational purposes only and should not be construed as limiting the subject matter. All or any portion of the documents cited herein, including but not limited to patents, patent applications, articles, books, operating manuals, and papers, are hereby incorporated by reference in their entirety.

[0057] The pharmaceutical composition according to the present invention comprises a compound of formula I or formula II according to the first aspect of the present invention, or an optical isomer, a pharma- ceutically acceptable salt or a prodrug thereof.

[0058] In this application, unless otherwise specified, the term "pharmaceutical acceptable salt" refers to a salt that is suitable for contact with a subject's tissue and does not cause undesirable side effects, and the salts in this application are mainly pharmaceutical acceptable acid addition salts and pharmaceutical acceptable base addition salts.

[0059] "Pharmaceutically acceptable acid addition salt" refers to a salt formed with an inorganic or organic acid which is capable of retaining the biological effectiveness of the free base without other adverse effects. Inorganic acid salts include hydrochlorides, hydrobromides, sulfates, nitrates, phosphates, etc., and organic acid salts include formates, acetates, 2,2-dichloroacetates, trifluoroacetates, propionates, caproates, caprylates, caprates, undecylenates, glycolates, gluconates, lactates, sebacates, adipates, glutarates, malonates, oxalates, maleates, succinates, fumarates, hydrochlorides, citrates, palmitates, stearates, oleates, cinnamates, laurates, malates, glutamates, pyroglutamates, aspartates, benzoates, methanesulfonates, benzenesulfonates, p-toluenesulfonates, alginates, ascorbates, salicylates, 4-aminosalicylates, naphthalenedisulfonates, etc. These salts can be produced by conventional methods known in the art.

[0060] "Pharmaceutically acceptable base addition salt" refers to a salt formed with an inorganic or organic base that can retain the biological effectiveness of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, aluminum salts, etc. Preferred inorganic salts are ammonium salts, sodium salts, potassium salts, calcium salts and magnesium salts. Salts derived from organic bases include primary amines, secondary amines, and tertiary amines, substituted amines include natural substituted amines, cyclic amines, and basic ion exchange resins, such as, but not limited to, ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosylamine, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like. Preferred organic bases include diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. These salts can be prepared by conventional methods known in the art.

[0061] The present invention further includes prodrugs of the above compounds, and in this application, the term "prodrug" refers to a compound that can be converted to the biologically active compound of the present invention under physiological conditions or by solvent decomposition. Thus, the term "prodrug" refers to a pharma- ceutically acceptable metabolic precursor of the compound of the present invention. When administered to an individual in need thereof, the prodrug may not have activity, but is converted to the active compound of the present invention in the body. Prodrugs are usually rapidly converted in the body, and the generation of the parent compound of the present invention is achieved, for example, by hydrolysis in blood. Prodrug compounds usually offer the advantage of solubility, tissue compatibility, or sustained release in the mammalian body. For specific methods of producing prodrugs, see Saulnier, MG, et al., Bioorg. Med. Chem. Lett. 1994, 4, 1985-1990; Greenwald, RB, et al., J. Med. Chem. 2000, 43, 475.

[0062] As used herein, the term "pharmaceutical acceptable" refers to a substance (e.g., a carrier or diluent) that does not affect the biological activity or properties of the compounds of the invention, and is relatively non-toxic, i.e., the substance may be administered to an individual without eliciting an adverse biological response or interacting in an inappropriate manner with any components included in the composition.

[0063] As used herein, the term "treatment" and other similar synonyms include: (i) preventing the disease or disorder from manifesting in a mammal, particularly where such mammal is susceptible to, but has not been diagnosed with, the disease or disorder; (ii) inhibiting the disease or disorder, i.e., inhibiting its progression; (iii) alleviating a disease or disorder, i.e., eliminating the symptoms of that disease or disorder; or (iv) It has the meaning of alleviating symptoms caused by the disease or disorder. [Brief description of the drawings]

[0064] [Figure 1] Effect of compound K18 on body weight in ICR mice, (A) is a single-dose experiment, (B) is a multiple-dose experiment. [Diagram 2] Histopathological changes in the liver, kidney and heart of mice treated with the vehicle group and with multiple doses (50 mg / kg) and single doses (500 mg / kg) of compound K18. Scale: 100 μM. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0065] Example 1: 2-[(2,2-dimethylpropyl){[4-(4-methylpiperazin-1-yl)phenyl]methyl}amino]pyrimidine-4-carbonitrile (K1) [ka] Compound M1 (500 mg, 2.47 mmol) was dissolved in 8 mL of anhydrous tetrahydrofuran, cooled to 0° C., and NaH (178 mg, 7.41 mmol) was added in batches while stirring, and the mixture was reacted at room temperature for 2 h. Compound M2 (732 mg, 2.72 mmol) was added while stirring, and the mixture was reacted at room temperature for 2 h. After the reaction was completed, 10 mL of water was added to quench the reaction, tetrahydrofuran was evaporated under reduced pressure, and the mixture was extracted twice with water. The organic phases were combined and dried over anhydrous sodium sulfate. Compound K1 was obtained as a white solid by column chromatography purification (petroleum ether: ethyl acetate = 20: 1) in a yield of 60.3%. 1 H NMR(400MHz,DMSO-d6) δ8.60(d,J=4.7Hz,1H),7.12(d,J=4.7Hz,1H),7.01(d,J=8.1Hz,2H),6.84(d,J=8.3Hz,2H),4.80( ESI-MS m / z:379.5[M+H] + .

[0066] Example 2: 2-[(2,2-dimethylpropyl)[(4-phenylphenyl)methyl]amino]pyrimidine-4-carbonitrile (K2) [ka] Intermediate M4 was prepared by referring to the synthesis method in Example 1.

[0067] Intermediate M4 (291 mg, 0.81 mmol) was dissolved in a mixed solution of 1 mL of water and 9 mL of DMF, and phenylboranediol (108 mg, 0.89 mmol) and potassium carbonate (223 mg, 1.616 mmol) were added in that order while stirring. The mixture was evacuated and heated to 80°C under argon gas protection for 4 hours. After the reaction was completed, the mixture was returned to room temperature, diluted with 20 mL of water, extracted twice with ethyl acetate, and the organic phases were combined, washed twice with saturated saline, and dried over anhydrous sodium sulfate. Compound K2 was obtained as a white solid by column chromatography (petroleum ether:ethyl acetate=12:1) in a yield of 54.7%. 1 H NMR(400MHz,DMSO-d6) δ8.65(s,1H),7.69-7.56(m,4H),7.45(q,J=7.2Hz,2H),7.36(t,J=6.7Hz,1H) ,7.30-7.15(m,3H),4.96(s,2H),3.58(s,2H),0.99(d,J=13.6Hz,9H);ESI-MS m / z:357.4[M+H] + .

[0068] Example 3: 2-[(2,2-dimethylpropyl)({4-[4-(methylthio)phenyl]phenyl}methyl)amino]pyrimidine-4-carbonitrile (K3) Referring to the synthesis method of Example 2, the raw material phenylborane diol was replaced with 4-methylsulfanylphenylborane diol to obtain compound K3 as a white solid with a yield of 48.6%. 1H NMR(400MHz,DMSO-d6) δ8.63(s,1H),7.63-7.55(m,4H),7.35-7.30(m,2H),7.23(d,J=7.9Hz,2H),7.17(d, ESI-MS m / z:403.4[M+H] + .

[0069] Example 4: 2-[(2,2-dimethylpropyl){[4-(4-fluorophenyl)phenyl]methyl}amino]pyrimidine-4-carbonitrile (K4) Referring to the synthesis method of Example 2, the raw material phenylboranediol was replaced with 4-fluorophenylboranediol to obtain compound K4 as a white solid, with a yield of 47.1%. 1 H NMR(400MHz,DMSO-d6) δ8.63(s,1H),7.62(d,J=20.0Hz,4H),7.49(s,2H),7.24(s,2H),7.17(s,1H),4.95(s,2H),3.58(s,2H),0.97(s,9H);ESI-MS m / z:375.4[M+H] + .

[0070] Example 5: 2-[(2,2-dimethylpropyl){[4-(4-methoxyphenyl)phenyl]methyl}amino]pyrimidine-4-carbonitrile (K5) Referring to the synthesis method of Example 2, the raw material phenylboranediol was replaced with 4-methoxyphenylboranediol to obtain compound K5 as a white solid, with a yield of 44.6%. 1 H NMR(400MHz,DMSO-d6) δ8.63(s,1H),7.55(t,J=8.7Hz,4H),7.20(d,J=7.8Hz,2H),7.16(d,J=4.7Hz,1H), 7.00(d,J=8.3Hz,2H),4.93(s,2H),3.78(s,3H),3.56(s,2H),0.97(s,9H);ESI-MS m / z:387.4[M+H] + .

[0071] Example 6: 2-[(2,2-dimethylpropyl)({4-[4-(methylsulfonyl)phenyl]phenyl}methyl)amino]pyrimidine-4-carbonitrile (K6) Referring to the synthesis method of Example 2, the raw material phenylboranediol was replaced with 4-methanesulfonylphenylboranediol to obtain compound K6 as a white solid, with a yield of 53.7%. 1 H NMR(400MHz,DMSO-d6) δ8.62(s,1H),7.98(d,J=8.5Hz,2H),7.90(d,J=8.2Hz,2H),7.69(d,J=7.9Hz,2H),7.29(d,J= 7.8Hz,2H),7.17(d,J=4.7Hz,1H),4.97(s,2H),3.59(s,2H),3.24(s,3H),0.97(s,9H);ESI-MS m / z:435.4[M+H] + .

[0072] Example 7: 2-({[4-(5-cyanothiophen-2-yl)phenyl]methyl}(2,2-dimethylpropyl)amino)pyrimidine-4-carbonitrile (K7) Referring to the synthesis method of Example 2, the raw material phenylboranediol was replaced with 5-(dihydroxyboryl)thiophene-2-carbonitrile to obtain compound K7 as a white solid with a yield of 31.7%. 1 H NMR(400MHz,DMSO-d6) δ8.62(s,1H),7.97(d,J=4.0Hz,1H),7.69(d,J=7.9Hz,2H),7.64(d,J=4.0Hz,1H),7.25 (d,J=7.9Hz,2H),7.17(d,J=4.7Hz,1H),4.93(s,2H),3.58(s,2H),0.96(s,9H);ESI-MS m / z:388.4[M+H] + .

[0073] Example 8: 2-({[4-(6-chloropyridin-3-yl)phenyl]methyl}(2,2-dimethylpropyl)amino)pyrimidine-4-carbonitrile (K8) Referring to the synthesis method of Example 2, the raw material phenylboranediol was replaced with (2-chloropyridin-5-yl)borondiol to obtain K8 compound as a white solid with a yield of 34.5%. 1 H NMR(400MHz,DMSO-d6) δ8.71(s,1H),8.63(s,1H),8.13(d,J=8.8Hz,1H),7.68(d,J=7.9Hz,2H),7.59(d,J=8.4Hz,1H ),7.28(d,J=7.8Hz,2H),7.18(d,J=4.7Hz,1H),4.96(s,2H),3.59(s,2H),0.97(s,9H);ESI-MS m / z:392.4[M+H] + .

[0074] Example 9: 2-({[4-(3,4-dichlorophenyl)phenyl]methyl}(2,2-dimethylpropyl)amino)pyrimidine-4-carbonitrile (K9) Referring to the synthesis method of Example 2, the raw material phenylboranediol was replaced with (1,2-dichlorobenzene-4-yl)borondiol to obtain K9 compound as a white solid with a yield of 52.9%. 1 H NMR(400MHz,DMSO-d6) δ8.62(s,1H),7.89(d,J=2.1Hz,1H),7.67(s,1H),7.65-7.61(m,3H),7.24(d,J= 8.0Hz,2H),7.16(d,J=4.7Hz,1H),4.94(s,2H),3.58(s,2H),0.96(s,9H);ESI-MS m / z:425.4[M+H] + .

[0075] Example 10: 2-[(2,2-dimethylpropyl){[4-(5-methylthiophen-2-yl)phenyl]methyl}amino]pyrimidine-4-carbonitrile (K10) Referring to the synthesis method of Example 2, the raw material phenylboranediol was replaced with (5-methylthiophen-2-yl)borondiol to obtain K10 compound as a white solid with a yield of 34.8%. 1H NMR(400MHz,DMSO-d6) δ8.62(s,1H),7.49(d,J=8.0Hz,2H),7.23(d,J=3.6Hz,1H),7.19-7.12(m,3H),6.79(dd, ESI-MS m / z:377.4[M+H] + .

[0076] Example 11: Methyl 4-(4-{[(4-cyanopyrimidin-2-yl)(2,2-dimethylpropyl)amino]methyl}phenyl)benzoate (K11) Referring to the synthesis method of Example 2, the raw material phenylboranediol was replaced with methyl 4-(dihydroxyboryl)benzoate to obtain K11 compound as a white solid, with a yield of 44.7%. 1 H NMR(400MHz,DMSO-d6) δ8.63(s,1H),8.01(d,J=8.4Hz,2H),7.79(d,J=8.4Hz,2H),7.68(d,J=7.9Hz,2H),7.27(d,J= 7.9Hz,2H),7.17(d,J=4.7Hz,1H),4.96(s,2H),3.87(s,3H),3.59(s,2H),0.97(s,9H);ESI-MS m / z:415.4[M+H] + .

[0077] Example 12: 2-({[4-(4-chloro-3-fluorophenyl)phenyl]methyl}(2,2-dimethylpropyl)amino)pyrimidine-4-carbonitrile (K12) Referring to the synthesis method of Example 2, the raw material phenylboranediol was replaced with (1-chloro-2-fluorobenzene-4-yl)borondiol to obtain K12 compound as a white solid with a yield of 51.8%. 1H NMR(400MHz,DMSO-d6) δ8.62(s,1H),7.71(dd,J=11.0,2.1Hz,1H),7.64(t,J=8.2Hz,3H),7.52(dd,J=8.4,2.1Hz,1H ),7.25(d,J=7.9Hz,2H),7.16(d,J=4.7Hz,1H),4.95(s,2H),3.58(s,2H),0.96(s,9H);ESI-MS m / z:409.4[M+H] + .

[0078] Example 13: 2-[(2,2-dimethylpropyl)({4-[4-(4-methylpiperazin-1-yl)phenyl]phenyl}methyl)amino]pyrimidine-4-carbonitrile (K13) Referring to the synthesis method of Example 2, the raw material phenylboranediol was replaced with 4-(4-methyl-1-piperazinyl)phenylboranediol to obtain K13 compound as a white solid with a yield of 68.7%. 1 H NMR(400MHz, CDCl3) δ8.42(d,J=4.7Hz,1H),7.47(d,J=8.1Hz,4H),7.17(s,2H),6.98(d,J=8.8Hz,2H),6.74(d,J=4.7Hz,1H ),4.99(s,2H),3.58(s,2H),3.29-3.23(m,4H),2.59(t,J=5.0Hz,4H),2.36(s,3H),1.01(s,9H);ESI-MS m / z:455.6[M+H] + .

[0079] Example 14: 2-[(cyclohexylmethyl)[(4-phenylphenyl)methyl]amino]pyrimidine-4-carbonitrile (K14) Referring to the synthesis method of Example 2, K14 compound was obtained as a white solid with a yield of 53.1%. 1H NMR(400MHz,DMSO-d6) δ8.65(s,1H),7.61(m,4H),7.44(m,2H),7.32(m,3H),7.15(m,1H),4.88(s,2 H),3.46(m,2H),1.81(s,1H),1.62(m,5H),1.14(m,3H),0.97(m,2H);ESI-MS m / z:383.5[M+H] + .

[0080] Example 15: 2-[(cyclohexylmethyl)({4-[4-(4-methylpiperazin-1-yl)phenyl]phenyl}methyl)amino]pyrimidine-4-carbonitrile (K15) Referring to the synthesis method of Example 2, K15 compound was obtained as a white solid with a yield of 60.7%. 1 H NMR(400MHz,DMSO-d6) δ8.63(s,1H),7.51(dd,J=15.1,8.1Hz,4H),7.23(d,J=7.7Hz,2H),7.14(d,J=4.7Hz,1H),6.99(d,J=8.4Hz,2H),4.85(s,2H) ),3.16(s,4H),2.45(s,4H),2.22(s,3H),1.81(s,1H),1.62(d,J=16.2Hz,5H),1.15(s,3H),0.97(d,J=11.8Hz,2H);ESI-MS m / z:481.7[M+H] + .

[0081] Example 16: 2-[(cyclohexylmethyl)({4-[4-(methylsulfonyl)phenyl]phenyl}methyl)amino]pyrimidine-4-carbonitrile (K16) Referring to the synthesis method of Example 2, K16 compound was obtained as a white solid with a yield of 49.3%. 1H NMR(400MHz,DMSO-d6) δ8.64(s,1H),7.98(d,J=8.5Hz,2H),7.91(d,J=8.3Hz,2H),7.70(d,J=7.8Hz,2H),7.35(d,J=7.9Hz,2H),7.15(d,J=4.7Hz,1H),4.90(s, ESI-MS m / z:461.6[M+H] + .

[0082] Example 17: 2-(Neopentyl(4-(thiazol-4-yl)benzyl)amino)pyrimidine-4-carbonitrile (K17) Referring to the synthesis method of Example 2, K17 compound was obtained as a white solid with a yield of 32.9%. 1 H NMR(400MHz,DMSO-d6) δ9.17(s,1H),8.62(s,1H),8.09(s,1H),7.91(s,2H),7.22(s,3H),4.93(s,2H),3.57(s,2H),0.96(s,9H);ESI-MS m / z:364.3[M+H] + .

[0083] Example 18: 2-((4-(2-(4-methylpiperazin-1-yl)thiazol-4-yl)benzyl)(neopentyl)amino)pyrimidine-4-carbonitrile (K18) By referring to the synthesis method of Example 2, a white solid K18 compound was obtained with a yield of 48.1%. 1 H NMR(400MHz,DMSO-d6) δ8.61(s,1H),7.75(t,J=8.6Hz,2H),7.26-7.07(m,4H),4.91(d,J=8.2Hz,2 H),3.56(s,2H),3.43(s,4H),2.42(s,4H),2.22(s,3H),0.95(s,9H);ESI-MS m / z:462.6[M+H] + .

[0084] Example 19 (1R,2R)-2-(8-fluoro-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-2-carbonyl)-N-(4-oxotetrahydrofuran-3-yl)cyclohexane-1-formamide (K19) [ka] Compound 1 (530 mg, 2.34 mmol) and compound 2 (300 mg, 1.95 mmol) were weighed and placed in a 100 mL eggplant flask, and 25 mL of DMF was added to dissolve the compound, followed by stirring at room temperature for 12 h. After completion of the reaction, the reaction solution was diluted with DCM, washed with 10% hydrochloric acid, dried over anhydrous sodium sulfate, and then separated by column chromatography (dichloromethane:methanol = 50:1) to obtain intermediate 3 as a yellowish white solid.

[0085] Intermediate 3 (100 mg, 0.29 mmol) and HATU (121 mg, 0.32 mmol) were weighed and placed in a 25 mL eggplant flask, 10 mL of acetonitrile was added, DIPEA (191 μL, 1.16 mmol) and compound 4 (33 mg, 0.32 mmol) were added, and the mixture was reacted for 10 min. After the reaction was completed, the mixture was dried and purified by column chromatography (dichloromethane:methanol = 50:1) to obtain intermediate 5 as a yellowish white solid.

[0086] The intermediate 5 was placed in a 25mL two-neck flask, protected with N2, and dissolved by adding 7mL of anhydrous DCM and DIPEA (386μL, 0.92mmol). The mixture was cooled to -15℃ in a low-temperature cold trap, and sulfur trioxide pyridine (146mg, 0.88mmol) was weighed and dissolved in 1mL of anhydrous DMSO to form a mixture, which was added to the reaction solution in the two-neck flask at -15℃ and reacted at low temperature for 1h. After the reaction was completed, the mixture was dried and purified by column chromatography (petroleum ether:ethyl acetate:methanol=16:4:1) to obtain compound K19 as a white solid product, with a yield of 68%. The MP146-148℃ was cooled to -15℃, and the mixture was cooled to -15℃ for 1h. 1H NMR(400MHz,DMSO-d6) δ10.98(s,1H),7.75(dd,J=16.0,8.3Hz,1H),7.39-7.09(m,2H),6.85(dtd,J=9.1,6.1,2.9Hz,1H),4.84-4.32(m,2H),4.1 4-3.73(m,3H),3.12-2.89(m,2H),2.86-2.63(m,2H),2.18(dt,J=17.4,7.6Hz,1H),1.97-1.54(m,7H),1.53-1.07(m,6H); 13 C NMR(101MHz,DMSO) δ215.55,175.04,173.87,170.82,158.33,156.03,135.42,132.87,126.01,112.14,108.84,106.89,103.17,102.58,60.23,56.09,49.0 7,46.42,43.18,42.55,41.83,41.43,39.37,35.69,30.44,29.57,29.16,25.70,24.61,23.49,21.22,18.18,14.55;MS(ESI):426.4[M+H] + .

[0087] Example 20 (1R,2R)-2-(2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide (K20) The preparation method of this embodiment refers to the preparation steps of Example 19, and replaces compound 1 with 2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indol-2-onium, and finally obtains compound K20 as a white solid with a yield of 71%. Its MP is 137-140°C, 1H NMR(400MHz,DMSO-d6) δ10.88(s,1H),8.30-8.01(m,1H),7.48(d,J=7.6Hz,0H),7.38(d,J=7.7Hz,1H),7.32-7.24(m,1H),7.08-6.91(m,2H) ),4.78-4.46(m,2H),4.26-3.52(m,7H),3.09-2.81(m,2H),2.74-2.54(m,2H),1.86-1.61(m,4H),1.40-1.14(m,4H); 13 C NMR(101MHz,CDCl3) δ211.38,211.31,175.85,175.79,174.54,174.16,136.00,131.28,125.61,121.72,119.67,117.50,110.85,106.62,70.07,69.9 0,69.71,69.49,54.76,54.69,46.33,46.17,43.39,42.96,40.07,39.87,29.27,28.86,25.27,24.49,23.29;MS(ESI):410.4[M+H] + .

[0088] Example 21 (1R,2R)-2-(8-fluoro-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-2-carbonyl)-N-((S)-4-oxotetrahydrofuran-3-yl)cyclohexane-1-formamide (K21) [ka] The preparation method of this example refers to the preparation steps of Example 19, in which compound 6 is used to replace compound 4, and compound K21 is obtained as a white solid product with a yield of 67%. 1 H NMR(400MHz,DMSO) δ11.01(s,1H),8.15-8.02(m,1H),7.32-7.14(m,2H),6.86(tdd,J=9.2,4.8,2.6Hz,1H),4.7 4-4.42(m,2H),4.34-3.38(m,7H),3.05-2.53(m,4H),1.92-1.57(m,4H),1.36-1.23(m,4H).

[0089] Example 22 (1R,2R)-2-(6,8-difluoro-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-2-carbonyl)-N-((S)-4-oxotetrahydrofuran-3-yl)cyclohexane-1-formamide (K22) The preparation method of this example refers to the preparation steps of Example 19, and the compound K22 is obtained as a white solid product with a yield of 66%. 1 H NMR(400MHz,DMSO) δ11.48(s,1H),8.30-8.02(m,1H),7.21-7.05(m,1H),6.88(d,J=12.6Hz,1H),4.70(q,J= 13.9Hz, 2H), 4.51-3.36 (m, 7H), 3.06-2.86 (m, 2H), 2.74-2.57 (m, 2H), 1.71-1.27 (m, 8H).

[0090] Example 23 (1R,2R)-2-(8-chloro-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-((S)-4-oxotetrahydrofuran-3-yl)cyclohexane-1-formamide (K23) The preparation method of this example refers to the preparation steps of Example 19, and the compound K23 is obtained as a white solid product with a yield of 65%. 1 H NMR(400MHz,DMSO) δ11.12(d,J=4.3Hz,1H),8.30-8.05(m,1H),7.63-7.42(m,1H),7.30(dd,J=8.6,1.9Hz,1H),7.03(ddt,J=7.9,3 .6,1.9Hz,1H),4.78-4.49(m,2H),4.48-3.39(m,7H),3.07-2.82(m,2H),2.79-2.57(m,2H),1.86-1.11(m,8H).

[0091] Example 24 (1R,2R)-2-(8-bromo-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-((S)-4-oxotetrahydrofuran-3-yl)cyclohexane-1-formamide (K24) The preparation method of this example refers to the preparation steps of Example 19, and compound K24 was obtained as a yellowish white solid with a yield of 69%. 1 H NMR(400MHz,DMSO) δ11.14(d,J=5.1Hz,1H),8.38-7.94(m,1H),7.80-7.51(m,1H),7.38-7.05(m,3H),4.88-4.43(m,2 H),4.35-3.45(m,7H),3.20-2.86(m,2H),2.87-2.58(m,2H),1.83-1.66(m,4H),1.36-1.19(m,4H).

[0092] Example 25 (1R,2R)-2-(6-(trifluoromethoxy)-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide (K25) The preparation method of this example refers to the preparation steps of Example 19, and the compound K25 is obtained as a white solid product with a yield of 40%. 1 H NMR(300MHz,DMSO) δ11.49(s,1H),8.32-8.05(m,1H),7.57-7.39(m,1H),7.04(d,J=5.4Hz,2H),4.83-4.55(m,2H),4.40 -3.44(m,7H),3.06-2.82(m,2H),2.78-2.54(m,2H),1.81(d,J=15.7Hz,4H),1.26(d,J=12.3Hz,4H).

[0093] Example 26 (1R,2R)-2-(6-(trifluoromethoxy)-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(S)-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide (K26) The preparation method of this example refers to the preparation steps of Example 19, and compound K26 was obtained as a white solid with a yield of 42%. 1H NMR(300MHz,DMSO) δ11.49(s,1H),8.29-8.03(m,1H),7.56-7.39(m,1H),7.04(d,J=5.3Hz,2H),4.82-4.64(m,2 H),4.57-3.39(m,7H),3.07-2.89(m,2H),2.78-2.54(m,2H),1.72(s,4H),1.33-1.23(m,4H).

[0094] Example 27 (1R,2R)-2-(6-fluoro-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(S)-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide (K27) The preparation method of this example refers to the preparation steps of Example 19, and compound K27 was obtained as a white solid with a yield of 66%. 1 H NMR(400MHz,DMSO) δ11.48(s,1H),8.30-8.05(m,1H),7.56-7.41(m,1H),7.04(d,J=6.5Hz,2H), 4.80-4.54(m,2H),4.54-3.38(m,7H),3.06-2.53(m,4H),1.91-1.25(m,8H).

[0095] Example 28 (1R,2R)-2-(6-fluoro-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(S)-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide (K28) The preparation method of this example refers to the preparation steps of Example 19, and compound K28 was obtained as a white solid with a yield of 51%. 1 H NMR(400MHz,DMSO) δ11.37(s,1H),8.34-8.04(m,1H),7.38-7.20(m,1H),6.96-6.88(m,2H),4.78-4.52(m, 2H), 4.51-3.44 (m, 7H), 3.08-2.90 (m, 2H), 2.81-2.57 (m, 2H), 1.73 (m, 4H), 1.25 (m, 4H).

[0096] Example 29 (1R,2R)-2-(6-Methoxy-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(S)-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide (K29) The preparation method of this example refers to the preparation steps of Example 19, and compound K29 ​​was obtained as a white solid with a yield of 63%. 1 H NMR(400MHz,DMSO) δ10.96(d,J=4.1Hz,1H),8.31-7.99(m,1H),7.12-6.83(m,2H),6.63(dd,J=7.6,5.7Hz,1H),4.77-4.40(m,2H) ,4.37-3.92(m,3H),3.89(s,3H),3.87-3.42(m,4H),3.04-2.54(m,5H),1.91-1.61(m,5H),1.35-1.13(m,4H).

[0097] Example 30 (1R,2R)-2-(6-Methoxy-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide (K30) The preparation method of this example refers to the preparation steps of Example 19, and compound K30 was obtained as a white solid with a yield of 43%. 1 H NMR(300MHz,DMSO) δ10.96(s,1H),8.33-8.02(m,1H),7.15-6.83(m,2H),6.63(dd,J=7.7,4.2Hz,1H),4.74-4.50(m,2H),4.4 9-3.88(m,7H),3.87-3.38(m,3H),3.13-2.76(m,2H),2.75-2.54(m,2H),1.91-1.72(m,4H),1.30(m,4H).

[0098] Example 31 (1R,2R)-2-(8-fluoro-6-methoxy-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(S)-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide (K31) The preparation method of this example refers to the preparation steps of Example 19, and the compound K31 is obtained as a white solid product with a yield of 68%. 1 H NMR(400MHz,DMSO) δ11.07(d,J=3.9Hz,1H),8.30-8.02(m,1H),6.94-6.72(m,1H),6.56(ddt,J=11.5,3.9,2.0Hz,1H),4.78-4 .46(m,2H),4.45-3.88(m,7H),3.87-3.46(s,3H),3.16-2.52(m,4H),2.05-1.45(m,4H),1.44-1.09(m,4H).

[0099] Example 32 (1R,2R)-2-(8-fluoro-6-methoxy-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide (K32) The preparation method of this example refers to the preparation steps of Example 19, and compound K32 was obtained as a white solid with a yield of 58%. 1 H NMR(400MHz,DMSO) δ11.07(s,1H),8.30-8.03(m,1H),6.91-6.75(m,1H),6.60-6.52(m,1H),4.71-4.45(m,2H),4 .45-3.88(m,7H),3.87-3.44(s,3H),3.04-2.75(m,2H),2.70-2.54(m,2H),1.82-1.26(m,8H).

[0100] Example 33 2-((4-chlorobenzyl)(neopentyl)amino)pyrimidine-4-carbonitrile (K33) The preparation method of this example refers to the preparation steps of Example 1. 4-chlorobenzyl bromide is used as the raw material to obtain a white solid product with a yield of 55%. 1 H NMR(400MHz,DMSO-d6) δ8.62(s,1H),7.36(d,J=8.4Hz,2H),7.21-7.15(m,3H),4.88(s,2H),3.56(s,2H),0.95(s,9H). 13 C NMR(101MHz,CDCl3) δ162.69,159.34,141.29,136.28,132.79,128.68,128.36,116.23,112.24,57.96,51.89,34.75,28.64.HRMS(ESI) for C 17 H 19 ClN4[M+H] +: calcd,315.1371;found,315.1370.

[0101] Example 34 2-((4-Bromobenzyl)(neopentyl)amino)pyrimidine-4-carbonitrile (K34) The preparation method of this example refers to the preparation steps of Example 1. 4-bromobenzyl bromide is used as the raw material to obtain a white solid product with a yield of 55%. 1 H NMR(400MHz,DMSO-d6) δ8.61(s,1H),7.48(d,J=8.2Hz,2H),7.17(d,J=4.7Hz,1H),7.11(d,J=8.0Hz,2H),4.85(s,2H),3.55(s,2H),0.94(s,9H). 13 C NMR(75MHz,CDCl3) δ162.68,159.35,141.29,136.82,131.63,128.84,120.85,116.23,112.26,57.98,51.96,34.75,28.63.HRMS(ESI) for C 17 H 19 BrN4[M+H] + :calcd,359.0866;found,359.0865.

[0102] Example 35 2-((Naphthalen-2-ylmethyl)(neopentyl)amino)pyrimidine-4-carbonitrile (K35) The preparation method of this example refers to the preparation steps of Example 1, and a white solid product is obtained with a yield of 50%. 1 H NMR(400MHz,DMSO-d6) δ8.60(s,1H),7.89-7.80(m,3H),7.62(s,1H),7.51-7.44(m,2H),7.34(d,J =8.5Hz,1H),7.17(d,J=4.7Hz,1H),5.08(s,2H),3.63(s,2H),0.98(s,9H). 13 C NMR(101MHz,CDCl3) δ162.90,159.35,141.32,135.21,133.33,132.64,128.40,127.70,126.1 9,125.72,125.29,116.33,112.11,57.91,52.50,34.81,28.71.HRMS(ESI) for C 21 H 22 N4[M+H] + :calcd,331.1917;found,331.1919.

[0103] Example 36 2-(Neopentyl(quinolin-2-ylmethyl)amino)pyrimidine-4-carbonitrile (K36) The preparation method of this example refers to the preparation steps of Example 1, and a white solid product is obtained with a yield of 27%. 1 H NMR(300MHz,DMSO-d6) δ8.75-8.44(m,1H),8.28(d,J=8.6Hz,1H),7.94-7.86(m,2H),7.77-7.66(m,1H),7.60-7.50 (m,1H),7.31(d,J=8.6Hz,1H),7.15(d,J=4.7Hz,1H),5.12(s,2H),3.71(s,2H),0.99(s,9H). 13C NMR(101MHz,CDCl3) δ162.93,159.29,158.69,147.93,141.24,136.59,129.57,129.02,127.59,12 7.21,126.15,118.62,116.28,112.29,59.41,55.82,34.71,28.58.HRMS(ESI) for C 20 H 21 N5[M+H] + :calcd,332.1870;found,332.1880.

[0104] Example 37 2-(((4'-(4-(2-hydroxyethyl)piperazin-1-yl)-[1,1'-biphenyl]-4-yl)methyl)(neopentyl)amino)pyrimidine-4-carbonitrile (K37) The preparation method of this example refers to the preparation steps of Example 1, and a white solid product is obtained with a yield of 53%. 1 H NMR(400MHz, CDCl3) δ8.45(d,J=4.7Hz,1H),7.56-7.45(m,4H),7.20(s,2H),7.00(d,J=8.8Hz,2H),6.77(d,J=4.7Hz,1H),5.02(s,2H),3.75(t, J=5.3Hz,2H),3.60(s,2H),3.38-3.27(m,4H),3.13(q,J=7.4Hz,2H),2.86-2.77(m,4H),2.72(t,J=5.3Hz,2H),1.03(s,9H). 13 C NMR(101MHz,CDCl3) δ162.87,159.34,150.14,141.26,139.61,135.89,132.25,127.67,127.30,126.64,116.33 ,116.29,112.03,77.39,59.58,57.57,52.90,52.03,48.64,45.91,34.76,28.68.HRMS(ESI) for C 29 H 36 NO[M+H] + :calcd,485.3023;found,485.3013.

[0105] Example 38 2-(((4'-(4-hydroxypiperidin-1-yl)-[1,1'-biphenyl]-4-yl)methyl)(neopentyl)amino)pyrimidine-4-carbonitrile (K38) The preparation method of this example refers to the preparation steps of Example 1, and a white solid product is obtained with a yield of 46%. 1 H NMR(300MHz,DMSO-d6) δ8.63(s,1H),7.56-7.43(m,4H),7.23-7.13(m,3H),7.03-6.93(m,2H),4.93(s,2H),4.71(d,J=4. 2Hz,1H),3.69-3.50(m,5H),2.95-2.81(m,2H),1.87-1.74(m,2H),1.54-1.39(m,2H),0.97(s,9H). 13 C NMR(101MHz,CDCl3) δ162.88,159.33,150.43,141.28,139.75,135.74,131.56,127.62,127.29,126.59, 116.51,116.34,112.02,67.88,57.70,52.03,47.07,34.77,34.09,28.69.HRMS(ESI) for C 28 H 33 NO[M+H] + :calcd,456.2758;found,456.2747.

[0106] Example 39 2-(((2-methyl-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)methyl)(neopentyl)amino)pyrimidine-4-carbonitrile (K39) The preparation method of this example refers to the preparation steps of Example 1, and a white solid product is obtained with a yield of 54%. 1H NMR(300MHz, CDCl3) δ8.46(d,J=4.7Hz,1H),7.23(d,J=8.7Hz,2H),7.18-7.13(m,1H),6.98(d,J=8.7Hz,4H),6.77(d,J=4.7Hz,1H ),5.00(s,2H),3.61(s,2H),3.35-3.25(m,4H),2.63(t,J=5.0Hz,4H),2.40(s,3H),2.27(s,3H),1.03(s,9H). 13 C NMR(101MHz,CDCl3) δ162.91,159.20,149.88,141.20,140.52,136.01,135.74,132.76,130.05,129.97,128.62,124 .12,116.35,115.39,111.97,57.80,55.15,51.90,48.85,46.13,34.78,28.71,20.74.HRMS(ESI) for C 29 H 36 N6[M+H] + :calcd,469.3074;found,469.3064.

[0107] Example 40 2-(((2-chloro-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)methyl)(neopentyl)amino)pyrimidine-4-carbonitrile (K40) The preparation method of this example refers to the preparation steps of Example 1, and a white solid product is obtained with a yield of 51%. 1 H NMR(300MHz, CDCl3) δ8.46(s,1H),7.36(d,J=8.2Hz,2H),7.28-7.19(m,2H),7.12-6.93(m,3H),6.80(d,J=4.6Hz, 1H),4.98(s,2H),3.61(s,2H),3.31(s,4H),2.62(s,4H),2.39(s,3H),1.03(s,9H).HRMS(ESI) for C 28 H 33 ClN6[M+H] + :calcd,489.2528;found,489.2519.

[0108] Example 41 2-(Neopentyl(4-(4-(2-oxopropyl)piperazin-1-yl)benzyl)amino)pyrimidine-4-carbonitrile (K41) [ka] Referring to the preparation steps of Example 1, a white solid product K41-1 was obtained, with a yield of 64%. 1 H NMR(400MHz,DMSO-d6) δ8.63(d,J=4.7Hz,1H),7.11(d,J=4.7Hz,1H),7.04(d,J=8.1Hz,2H),6.91(d,J=8.1Hz,2H),4 .81(s,2H),3.44(s,2H),3.37(t,J=4.9Hz,4H),3.14-3.04(m,4H),1.42(s,9H),0.94(s,9H).

[0109] Intermediate K41-1 (330 mg, 1.41 mg) was added to 10 mL of cold 4N hydrochloric acid / dioxane solution and reacted in an ice bath for 30 minutes. After detecting the end of the reaction by TLC, the reaction solution was evaporated under reduced pressure, and a small amount of toluene was added until the reaction solution disappeared. The residue was then dissolved in 10 mL of DMF, potassium carbonate (487 mg, 3.53 mmol) and chloroacetone (195 mg, 2.11 mmol) were added in sequence, and the reaction solution was heated to 80 ° C. and kept warm for 2 hours to react. After detecting the end of the reaction by TLC, it was cooled to room temperature. 20 mL of water was added to the reaction solution, extracted with ethyl acetate (20 mL x 3), and the organic phase was combined and washed with dilute hydrochloric acid (10 mL x 2) and saturated sodium chloride solution (20 mL x 2) in sequence, and then dried over anhydrous sodium sulfate for 4 hours. After suction filtration and concentration of the filtrate under reduced pressure, column chromatography purification (petroleum ether:ethyl acetate=1:1) was performed to give 249 mg of a pale yellow solid K41, with a total yield of 42% for the two steps. 1H NMR(400MHz,DMSO-d6) δ8.61(d,J=4.7Hz,1H),7.13(dd,J=4.7,1.6Hz,1H),7.02(d,J=8.4Hz,2H),6.85(d,J=8.3Hz,2H),4.81(s,2H),3 .52(s,2H),3.28-3.21(m,2H),3.16-3.03(m,4H),2.59-2.52(m,4H),2.16-2.02(m,3H),0.95(s,9H).HRMS(ESI) for C 24 H 32 NO[M+H] + :calcd,421.2710;found,421.2704.

[0110] Example 42 2-(isobutyl((4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)methyl)amino)pyrimidine-4-carbonitrile (K42) Referring to the preparation steps of Example 1, a white solid product was obtained, with a yield of 46%. 1 H NMR(300MHz,DMSO-d6) δ8.66(s,1H),7.59-7.46(m,4H),7.25(d,J=7.9Hz,2H),7.17(d,J=4.6Hz,1H),7.00(d,J=8.6Hz,2H),4.88(s,2H) ,3.47-3.40(m,2H),3.23-3.12(m,4H),2.49-2.42(m,4H),2.23(s,3H),2.19-2.08(m,1H),0.88(d,J=6.6Hz,6H). 13 C NMR(101MHz,CDCl3) δ162.18,159.56,150.53,141.56,139.92,135.68,131.81,127.95,127.65,126.64, 116.40,116.05,111.87,55.10,54.33,50.61,48.86,46.20,26.78,20.24.HRMS(ESI) for C 27 H 32 N6[M+H] + :calcd,441.2761;found,441.2753.

[0111] Example 43 N-(4-((4-cyanopyrimidin-2-yl)(neopentyl)amino)methyl)phenyl)-2-(4-methylpiperazin-1-yl)acetamide (K43) Referring to the preparation steps of Example 1, a white solid product was obtained, with a yield of 43%. 1 H NMR(300MHz, CDCl3) δ9.09(s,1H),8.43(d,J=4.7Hz,1H),7.50(d,J=8.5Hz,2H),7.22-7.08(m,2H),6.76(d,J=4.7 Hz,1H),4.94(s,2H),3.56(s,2H),3.15(s,2H),2.79-2.41(m,8H),2.36(s,3H),1.01(s,9H). 13 C NMR(101MHz,CDCl3) δ168.12,162.76,159.26,141.22,136.45,133.68,127.80,119.67,116.29, 112.03,61.72,57.71,54.99,52.95,51.98,45.63,34.72,28.64.HRMS(ESI) for C 24 H 33 NO[M+H] + :calcd,436.2819;found,436.2811.

[0112] Example 44 2-((4-(4-methylpiperazine-1-carbonyl)benzyl)(neopentyl)amino)pyrimidine-4-carbonitrile (K44) [ka] Intermediate K44-1 was dissolved in 10 mL of acetonitrile, and DIPEA (2.13 g, 16.48 mmol) was added, and the reaction solution was heated to 80° C. and reacted for 16 hours. The end of the reaction was detected by TLC, and then cooled to room temperature. 20 mL of water was added to the reaction solution, and the solution was extracted with ethyl acetate (20 mL x 3). The organic phases were combined and washed with dilute hydrochloric acid (10 mL x 2) and saturated sodium chloride solution (20 mL x 2) in that order, and then dried over anhydrous sodium sulfate for 4 hours. The mixture was filtered by suction, and the filtrate was concentrated under reduced pressure, and then purified by column chromatography (petroleum ether: ethyl acetate = 12: 1) to obtain white solid K44-2, with a yield of 55%. 1 H NMR(300MHz,DMSO-d6) δ8.59(s,1H),7.89(d,J=8.2Hz,2H),7.28(d,J=8.0Hz,2H),7.18(d,J=4.7Hz,1H),4.96(s,2H),3.83(s,3H),3.60(s,2H),0.96(s,9H).

[0113] Intermediate K44-2 (700 mg, 2.07 mmol) was added to 8 mL of a mixed solution of methanol and water (3:1), lithium hydroxide (99 mg, 4.14 mmol) was added, and then the mixture was reacted in an ice bath for 4 hours. After detecting the completion of the reaction by TLC, the reaction solution was concentrated, and then the pH was adjusted to about 2 with 1N hydrochloric acid solution in an ice bath to produce a large amount of white solid, which was suction filtered, the filter cake was washed with ethyl acetate and a small amount of ethanol, and dried overnight in a vacuum environment to obtain a crude carboxylic acid product, which was used directly in the next step of the reaction. The obtained carboxylic acid was added to 10 mL of anhydrous tetrahydrofuran, and HATU (787 mg, 2.07 mmol), triethylamine (524 mg, 5.18 mmol) and N-methylpiperazine (249 mg, 2.48 mmol) were added in order, and the mixture was reacted at room temperature for 2 hours. After detecting the completion of the reaction by TLC, the reaction solution was diluted with 40mL of ethyl acetate, washed with saturated sodium carbonate solution (15mL x 2), dilute hydrochloric acid solution (15mL x 2) and saturated sodium chloride solution (15mL x 2) in sequence, and then dried over anhydrous sodium sulfate for 4 hours. After suction filtration and concentrating the filtrate under reduced pressure, column chromatography purification (petroleum ether: ethyl acetate = 1:2) was performed to obtain 480mg of white solid of compound K44, the yield of the two steps was 57%. 1 H NMR(300MHz, CDCl3) δ8.44(s,1H),7.35(d,J=7.8Hz,2H),7.18(d,J=7.8Hz,2H),6.79(d,J=4.7 Hz,1H),4.99(s,2H),3.83(s,2H),3.57(s,4H),2.37(s,7H),1.01(s,9H). 13 C NMR(101MHz,CDCl3) δ170.13,162.74,159.36,141.25,139.65,134.38,127.39,127.24,126.98,116.22 ,112.25,58.07,55.16,54.67,52.19,47.50,45.91,41.93,34.71,28.62.HRMS(ESI) for C 23 H 30 NO[M+H] + :calcd,407.2554;found,407.2547.

[0114] Example 45 4-(((4-cyanopyrimidin-2-yl)(neopentyl)amino)methyl)-N-(2-(4-methylpiperazin-1-yl)-2-oxoethyl)benzamide (K45) Referring to the preparation steps of Example 44, a white solid product was obtained, and the yield was 34%. 1 H NMR(300MHz,DMSO-d6) δ8.72-8.47(m,2H),7.77(d,J=8.0Hz,2H),7.23(d,J=8.0Hz,2H),7.18(d,J=4.7Hz,1H),4.94(s,2H) ),3.59(s,2H),3.18-3.02(m,6H),2.95-2.80(m,2H),2.70(s,3H),1.89-1.69(m,3H),0.96(s,9H). 13 C NMR(101MHz,CDCl3) δ167.71,162.65,159.41,141.54,141.10,132.81,127.62,126.93,116.25,112.29 ,58.21,54.59,52.36,45.88,44.51,43.65,34.67,33.18,28.56,27.22.HRMS(ESI) for C 25 H 34 NO[M+H] + :calcd,435.2867;found,435.2859.

[0115] Example 46 Inhibitory effect of compounds on Cat K enzyme Reagent information: Cat K inhibitor screening kit: Lot number, 6L23K01500; Supplier, BiVision. Drug preparation: Compounds were dissolved in DMSO and prepared into 10 mM stock solutions. When used, prepare the desired concentration with Buffer.

[0116] Ref-03 is the comparative compound, and the general formula is: [ka] It is.

[0117] Experimental Method 1. Primary drug screening Cathepsin K (CTSK, EC 3.4.22.38) is a lysosomal cysteine ​​protease that is involved in osteoclast bone remodeling and resorption and can further degrade collagen, gelatin and elastin. Biovision's Cathepsin K Inhibitor Screening Kit utilizes the ability of active Cathepsin K to cleave a synthesized AFC-based peptide substrate to release AFC, which can be easily quantified using a fluorometer or fluorescence microplate reader. In the presence of a Cathepsin K-specific inhibitor, reduced / elimination of the degradation of the substrate causes a decrease or complete loss of AFC fluorescence. Such a simple, high-throughput, adaptable assay kit can be used to screen / research / characterize potential inhibitors of Cathepsin K.

[0118]

number

[0119] 20μL of buffer, Cat K inhibitor, and test compound (1μM, 10μM) were added to a 96-well plate to form EC well, IC well, and S well, respectively, and 50μL of Cat K Enzyme Solution was added to each well and incubated at room temperature for 10-15min to construct the enzyme-inhibitor complex, and 30μL of Cat K Substrate Solution was added to each well and incubated at room temperature for 30-60min, and the final reaction volume was 100μL. At 30-60min, two time points T1 and T2 were selected to detect the fluorescence absorption value (Ex / Em=400 / 505nm), and the fluorescence absorption value was defined as RFU1 and RFU2, and the inhibition rate (%) of the test compound against Cat K enzyme was calculated.

[0120] Slope = (RFU2-RFU1) / (T2-T1) Inhibition rate (%) = (EC slope - S slope) / EC slope x 100 2.IC 50 measurement The screening method is the same as 1.1. The concentration of the test compound is set between 0.1 nM and 10 μM, and 4-5 concentrations are selected for detection and IC 50 Draw a curve and IC 50 values ​​were calculated.

[0121] Data processing: All data were statistically analyzed using Graph pad.

[0122] Experimental Results The results are shown in Table 1.

[0123] [Table 1]

[0124] The compounds of the present invention have high inhibitory rates against Cat K enzyme and IC 50 The IC values ​​of some of the compounds, e.g., K13 and K15, were low and showed good inhibitory activity against the Cat K enzyme. 50 The IC value is less than 10 nM, and the IC of the other parts of the compound, e.g., K1, K18-K22, 50 All of the values ​​were less than 100 nM, and therefore the compounds of the present invention have good inhibitory effects on the Cat K enzyme.

[0125] Example 47 Inhibitory effect of compounds on Cat B enzyme and Cat S enzyme Cathepsin B Screening 10 μL of buffer, Cat B inhibitor, and test compound (1 μM, 10 μM) were added to a 96-well plate to form EC well, IC well, and S well, respectively, and 50 μL of Cat B Enzyme Solution was added to each well and incubated at room temperature for 10-15 min to construct the enzyme-inhibitor complex, and 40 μL of Cat B Substrate Solution was added to each well and incubated at room temperature for 30-60 min, and the final reaction volume was 100 μL. At 30-60 min, two time points T1 and T2 were selected to detect the fluorescence absorption value (Ex / Em=400 / 505 nm), and the fluorescence absorption value was defined as RFU1 and RFU2, and the inhibition rate (%) of the test compound against Cat B enzyme was calculated.

[0126] Slope = (RFU2-RFU1) / (T2-T1) Inhibition rate (%) = (EC slope - S slope) / EC slope x 100

[0127] I C 50 measurement The screening method was the same as above, and the concentration of the test compound was set between 0.1 nM and 10 μM, and 4-5 concentrations were selected for detection, and the IC 50 Draw a curve and IC 50 values ​​were calculated.

[0128] Data processing: All data were statistically analyzed using Graph pad.

[0129] Cathepsin S screening 10 μL of buffer, Cat S inhibitor, and test compound (1 μM, 10 μM) were added to a 96-well plate to form EC well, IC well, and S well, respectively, and 50 μL of Cat B Enzyme Solution was added to each well and incubated at room temperature for 10-15 min to construct the enzyme-inhibitor complex, and 40 μL of Cat S Substrate Solution was added to each well and incubated at room temperature for 30-60 min, and the final reaction volume was 100 μL. At 30-60 min, two time points T1 and T2 were selected to detect the fluorescence absorption value (Ex / Em=400 / 505 nm), and the fluorescence absorption value was set as RFU1 and RFU2, and the inhibition rate (%) of the test compound against Cat S enzyme was calculated.

[0130] Slope = (RFU2-RFU1) / (T2-T1) Inhibition rate (%) = (EC slope - S slope) / EC slope x 100

[0131] I C 50 measurement The screening method was the same as above, and the concentration of the test compound was set between 0.1 nM and 10 μM, and 4-5 concentrations were selected for detection, and the IC 50 Draw a curve and IC 50 values ​​were calculated.

[0132] Data processing: All data were statistically analyzed using Graph pad.

[0133] The results of the ratios of the inhibitory activities of the compounds against Cat S, Cat B, and Cat K are shown in Table 2.

[0134] [Table 2]

[0135] Some of the above compounds show selectivity for Cat K far more than 1000 times higher than the other two enzymes. In this experiment, the test value of the positive control Odanacatib is slightly lower than that reported in the literature. However, in comparison, the Cat K inhibitor of the present invention still significantly improves the selectivity for Cat K, especially compensating for its lack of selectivity for Cat K and Cat S.

[0136] Example 48 Intrinsic clearance and half-life of compounds in rat liver microsomes 1. Sample Solution Preparation Centrifuge tubes containing the test substance powders K2 (molecular formula C23H27N3O4, 4.6 mg), K3 (molecular formula C23H26FN3O4, 5.0 mg), and K4 (molecular formula C23H25F2N3O4, 5.6 mg) were taken, and 1 mL of dichloromethane was added to each to dissolve them. After ultrasonic treatment for 30 minutes, K2 stock solution with mass concentrations of 4.6 mg / mL, K3 stock solution with mass concentrations of 5.0 mg / mL, and K4 stock solution with mass concentrations of 5.6 mg / mL were obtained. These were sealed and stored in a refrigerator at -20°C until ready for use. Just before use, the samples were serially diluted with methanol to the corresponding mass concentrations to obtain K2 series standard sample working solutions of 0.02, 0.05, 0.22, 0.49, 0.96, 2.37, and 4.90 ng / mL, K3 series standard sample working solutions of 0.02, 0.05, 0.10, 0.19, 1.00, 2.01, and 4.93 ng / mL, and K4 series standard sample working solutions of 0.10, 0.20, 0.53, 0.96, 2.42, 5.10, and 10.10 ng / mL, respectively.

[0137] 2. Test Method Using a pipette, 5μL of the test drug, 2μL of rat liver microsomes, and 468μL of PBS buffer were taken to prepare a premix solution, which was then incubated in a water bath at 37℃ for 5min. After the incubation was completed, 47.5μL of premix solution, 100μL of ice-cold acetonitrile, and 2.5μL of PBS buffer were taken into another new tube and gently mixed evenly to obtain the control group. The remaining premix solution was added with 22.5μL of NADPH solution to start the reaction, lightly blown, and immediately 50μL of premix solution and 100μL of acetonitrile were taken into another new tube to end the reaction. The incubation solution was then placed in a water bath pot and incubated, and 50μL was taken at 5min, 10min, 20min, 30min, 45min, 60min, and 90min, respectively, and 100μL of acetonitrile was added to end the reaction, and the mixture was centrifuged at 12000r / min for 10min, and the supernatant was taken and detected by the equipment.

[0138] Chromatographic conditions Chromatography column: Hypersil GOLD 100*2.1mm 3 μm liquid chromatography column Mobile phase: 0.1% formic acid in water (A) 100% acetonitrile (B) Gradient elution (0-1 min: 50% B, 1-5 min: 50% B-90% B, 5-7 min: 90% B, 7-7.1 min: 50% B, 7.1-10 min: 50% B) Flow rate: 0.3 ml / min Column temperature: 40°C Sample supply amount: 5 μL Mass spectrometry conditions Electrospray ion source (ESI); ion spray voltage: 4500 V, ion source temperature: 450° C., air curtain air pressure: 35 psi, and declustering potential was 80 V for both.

[0139] 3. Preparation of calibration curve and consideration of lower limit of quantification The sample solution was analyzed by supplying samples under the above chromatography and mass spectrometry conditions, and the peak areas were recorded. A linear regression was performed using the weighting method (weighting coefficient is 1 / x2) with the mass concentration of the test substance as the horizontal axis (x) and the peak area of ​​the test substance as the vertical axis (y). The results showed that the linear range of K2 mass concentration detection was 0.02-4.90 ng / mL, with a lower limit of quantification of 0.02 ng / mL, the linear range of K3 mass concentration detection was 0.02-4.93 ng / mL, with a lower limit of quantification of 0.02 ng / mL, and the linear range of K4 mass concentration detection was 0.10-10.10 ng / mL, with a lower limit of quantification of 0.10 ng / mL. Statistical analysis was performed from the results to determine the clearance (Cl int ) and half-life (T 1 / 2 ) was calculated.

[0140] The results are shown in Table 3. [Table 3]

[0141] The above compounds all have lower intrinsic clearance in rat liver microsomes than Ref-03, among which the clearance of compound K4 is 2 times lower, and the clearance of other compounds is smaller than that of Ref-03. As for half-life, the half-life of Ref-03 is 0.6 hours, while the half-life of the compounds of the present invention is longer than 0.6 hours. Therefore, the metabolic stability of the compounds of the present invention is higher.

[0142] Example 49 Inhibitory effect of compounds on osteoclasts 1. Detection of cell activity (toxicity) 1 x 10 RAW264.7 cells 5The cells were seeded in a 96-well plate at a density of 100 μL / well and cultured in DMEM medium containing 10% FBS and 1% penicillin / streptomycin. The cells were cultured in a constant temperature incubator at 37°C and 5% CO2. After 24 hours of culture, different concentrations of inhibitor working solution were added to the treatment group, and an equal volume of PBS working solution was added to the control group. After continued culture for 2 days, the culture supernatant was removed, washed three times with PBS, and the cell viability was detected using a CCK8 cell proliferation detection kit. 100 μL of CCK8-containing PBS was added to each well, and incubated at 37°C for 2 hours in the dark, after which the absorbance at 450 nm was detected using a microplate reader. CCK8 was added to the medium without cells or drugs to create a blank group.

[0143] The survival rate was calculated according to the following formula: Cell viability (%)=[(mean absorbance of treatment group−mean absorbance of blank wells) / (mean absorbance of control group−mean absorbance of blank group)]×100.

[0144] The results are shown in Table 4. [Table 4]

[0145] As can be seen from Table 5, the cell viability of RAW264.7 cells in the compound K29 ​​at concentrations of 1 μM and 10 μM is 96.06% and 98.89%, respectively, and the cell viability of RAW264.7 cells in the compound K31 at concentrations of 1 μM and 10 μM is 85.84% and 82.99%, respectively. As can be seen, compound K29 ​​has no obvious cytotoxicity to RAW264.7 cells at concentrations of 1 μM and 10 μM, and compound K31 has cytotoxicity to RAW264.7 cells at concentrations of 1 μM and 10 μM.

[0146] 2. Osteoclast differentiation RAW264.7 cells were cultured in a 96-well plate at 1 x 10 5The cells were seeded at a density of 100 / well and cultured in DMEM medium containing 10% fetal bovine serum and 1% penicillin / streptomycin. The cells were cultured in a constant temperature incubator at 37°C and 5% CO2. 100ng / mL RANKL induction solution was added to each well in the treatment group and model group to culture osteoclasts, and the solution (containing RANKL) was replaced every 2-3 days. After the third liquid replacement, inhibitor working solutions of different concentrations were added to the treatment group, and an equal volume of PBS working solution was added to the model group and cultured for 24h. The control group was added with the same total volume of PBS working solution without adding RANKL induction solution or inhibitor working solution. The culture supernatant was removed, washed three times with PBS, and the cells were stained with a TRAP staining kit. Multinuclear osteoclasts were observed and counted under an optical microscope.

[0147] The results are shown in Table 5. [Table 5]

[0148] As can be seen from Table 5, compared with the model group, compound K29 ​​at concentrations of 1 μM and 10 μM and K31 at concentrations of 100 nM and 500 nM all tend to inhibit osteoclast formation, among which, K29 significantly inhibits osteoclast formation at a concentration of 10 μM. Referring to Tables 4 and 5, compound K29 ​​has no cytotoxicity and has good effect of inhibiting osteoclast formation.

[0149] Example 50 In vivo toxicity studies The experimental subjects were male ICR mice aged 6-8 weeks. After one week of adaptation, they were randomly divided into groups according to body weight. Different concentrations of the compounds were then prepared and administered intraperitoneally (ip) in single and multiple dose experiments, and the vehicle in the same group was used as a reference.

[0150] As shown in Figure 1, when the single dose was 500mg / kg and 300mg / kg, the mice were observed on the 10th day, and no deaths or obvious toxic reactions occurred. During that time, the mice were weighed daily, and no significant difference was observed in the weight of the treated group compared to the vehicle group (P>0.05). In the multiple-dose experiment, the doses of 50mg / kg and 25mg / kg were selected and administered once a day. After one week of continuous administration, the mice were observed, and no deaths or obvious toxic reactions occurred. Compared with the vehicle group, the weights of the treated groups showed no significant difference (P>0.05). As shown in Figure 2, histopathological studies were performed on the mice in the single-dose and multiple-dose maximum dose groups, and no obvious pathological changes were observed in the mice treated with compound K18 compared to the vehicle group. The safe doses of 50mg / kg and 25mg / kg meet the safety requirements for subsequent in vivo pharmacodynamic studies.

[0151] The above results demonstrate that compound K18 has low toxicity in the body and a certain degree of safety.

[0152] Example 51 Anti-osteoporosis in vivo drug efficacy experiment Dexamethasone was subcutaneously injected into male C57BL / 6 mice to establish an osteoporosis mouse model. After 8 days of model establishment, the model mice were intraperitoneally injected with 12mg / kg, 24mg / kg, and 36mg / kg doses, and the Cat K inhibitor Balicatib was used as a positive control. After daily administration for 6 weeks, the mice were anesthetized and dissected, and serum was preserved. The content of the bone resorption biomarker CTX in the blood was detected by ELISA, and one side of the femur was taken to measure bone density. The therapeutic effect of the Cat K inhibitor on osteoporosis was evaluated by comparing with the vehicle group. The effect of the preliminary experiment was obvious. Now, the formal experiment is proceeding.

[0153] Example 52: Anti-bone metastasis tumor in vivo drug efficacy experiment The therapeutic effect of the compound of the present invention on bone metastasis tumors was further studied by establishing a prostate cancer bone metastasis model by injecting C4-2B cells into the tibia of NOD-SCID mice. Four weeks after the model was established, the model mice were intraperitoneally injected with 25 mg / kg and 50 mg / kg doses, and zoledronic acid, a common anti-bone metastasis drug, was used as a positive control. The compound was administered once a day for four consecutive weeks, during which the changes in the body weight and tumor volume of the mice were observed and recorded. After the administration was completed, the mice were killed and the content of prostate specific antigen (PSA) in the plasma was measured by ELISA, and then the bone density of the mouse tibia was measured and pathological sections were prepared to measure the pathological changes in the bone tissue. Based on these test results, the inhibitory effect of Cat K inhibitor on the progression of prostate cancer bone metastasis tumors and the protective effect on bone tissue were evaluated.

[0154] The test results preliminary demonstrated that compound K18 could inhibit the migration, invasion and adhesion processes of C4-2B cells in a concentration-dependent manner, indicating that Cat K inhibitors could effectively exert antitumor activity by affecting the tumor metastasis process.

[0155] Example 53 Absolute bioavailability experiment by oral administration to rats 1 equipment Acquity UPLC ultra-high efficiency liquid chromatography system (Waters), TSQ Endura triple quaternary rod MS (Thermo), KQ-500E ultrasonic cleaner (Kunshan Ultrasonic Equipment Co., Ltd.), 3K15 tabletop high-speed centrifuge (SIGMA, Germany), XW-80A vortex mixer (Shanghai Precision Industry Co., Ltd.) 2. Chemicals and Reagents Test target substance: K29 (molecular weight 439.21, mass 27 mg), K31 (molecular weight 457.20, mass 14 mg + 20 mg).

[0156] Reagents: HS-15 was an analytical reagent, sodium carboxymethylcellulose (CMC-Na) was pharmaceutical grade, formic acid, methanol, and acetonitrile were all chromatographic grade, and water was purified water.

[0157] 3. Test Method 3.1 Chromatography and mass spectrometry conditions 3.1.1 Chromatographic conditions Chromatography column: Thermo Scientific 100*2.1mm 3μm liquid chromatography column Mobile phase: 5 mM ammonium acetate in water (A), 100% acetonitrile (B) Gradient elution (0-0.5min: 20%B, 0.5-1.5min: 20%-90%B, 1.5-4min: 90%B, 4-4.1min: 90%-20%B, 4.1-4.5min: 20%B) Flow rate: 0.25mL / min Column temperature: 40℃ Sample supply amount: 2μL 3.1.2 Mass spectrometry conditions Electrospray ion source (ESI); ion spray voltage: 3500 V, ion source temperature: 300° C., auxiliary gas: 45 Arb, sheath gas 10 Arb.

[0158] 3.2 Preparation of dosing solutions Intravenous intraperitoneal administration solution: Prepared in 0.5% HS-15 saline solution, concentration is 0.2 mg / mL. Suspension for intragastric administration: The sample is ground and suspended in a 0.5% aqueous CMC-Na solution to give a concentration of 0.5 mg / mL.

[0159] 3.3 Animal treatment and sample processing 3.3.1 Administration and blood sampling Each administration method for each compound was tested using two SD rats.

[0160] The two compounds K29 and K31 were administered intravenously and intragastrically. The intravenous administration volume was 5mL / kg, and 0.2mL of blood was collected from the jugular vein at 2min, 10min, 30min, 1h, 2h, 4h, 7.5h, and 24h after administration. The intragastric administration volume was 10mL / kg, and 0.2mL of blood was collected from the jugular vein at 5min, 15min, 30min, 1h, 2h, 4h, 8h, and 24h after administration. All blood samples were anticoagulated with heparin and centrifuged at 4000g for 10min. The supernatant plasma was collected and frozen for testing.

[0161] 3.3.2 Sample processing 50 μL of plasma sample was taken, 150 μL of acetonitrile was added, mixed uniformly by vortexing, centrifuged at 9100 g for 10 min, and 2 μL of the supernatant was taken as a sample to be detected by the equipment.

[0162] The test results are shown in Table 6-10. [Table 6]

[0163] [Table 7]

[0164] [Table 8]

[0165] [Table 9]

[0166] [Table 10]

[0167] As can be seen from Table 10, the oral bioavailabilities of compounds K29 and K31 were 62.1% and 51.0%, respectively, and the oral bioavailability was good.

Claims

1. Formula 0 containing a structure represented by the following general formula: 【Chemistry 1】 [In the formula, X is C or N; R １ is selected from H, a substituted or unsubstituted C1-10 alkyl group, and a substituted or unsubstituted C3-C10 cycloalkyl group, wherein the substituent is selected from a halogen, an amino group, a cyano group, a hydroxy group, an aldehyde group, a carboxyl group, and a sulfone group; R ＊ is selected from halogen or a C1-6 alkyl group; Y is a cyclic group, which is located at any position of the aromatic ring to which it is connected, and which is bonded to the aromatic ring via one or two carbon atoms, and which may be a C3-10 cycloalkyl group, a C6-12 aromatic ring, or a C5-12 heterocycle; the C6-12 aromatic ring includes a C6-12 aromatic ring and a C6-12 aromatic heterocycle, the C6-12 aromatic heterocycle contains at least one heteroatom, the C5-12 heterocycle is a saturated heterocycle or an unsaturated heterocycle, the heterocycle contains 1-3 heteroatoms, and the heteroatom may be O, N or S; t is 0 or 1, and when t is 0, R ５ is located at any position on the aromatic ring to which it is bonded, R ５ represents H, halogen, an amino group, a cyano group, a C1-10 alkyl group, a C1-10 alkoxy group, a C3-10 cycloalkyl group, a substituted C3-10 heterocycloalkyl group, -S(O) ２ R ２ , -C(O)R ２ , -NR ３ R ４ , —C(O)NHR ７ , -SR ６ , -OR ６ is selected from the substituents of the C3-10 heterocycloalkyl group are selected from a hydroxy group and a C1-10 alkyl group; The C1-10 alkyl group may further include a hydroxy group, a —C(O)R ２ may be substituted with The R ２ is selected from H, an amino group, a halogen atom, a substituted or unsubstituted C1-6 alkyl group, a C1-6 alkoxy group, a C3-8 cycloalkyl group, or a C3-8 heterocycloalkyl group, and the substituent of the C1-6 alkyl group, C1-6 alkoxy group, C3-8 cycloalkyl group, or C3-8 heterocycloalkyl group is a C1-6 alkyl group; The R ３ , R ４ are each independently H, —C(O)R ８ or R ３ , R ４ form a 4-8 membered ring containing at least one N together with the N to which they are attached, and ８ is selected from a C1-6 alkyl group substituted with a piperazinyl group or a methylpiperazine group; R ６ is selected from C1-6 alkyl groups; R ７ is selected from a C alkyl group, and the C alkyl group is further selected from -C(O)R ９ and R ９ is selected from a piperazinyl group or a methylpiperazine group, the C3-10 heterocycloalkyl group or the C3-8 heterocycloalkyl group contains 1-3 heteroatoms, which may be O, N or S; The halogen may be mono- or polysubstituted and is selected from F, Cl, Br, I. or a pharmaceutically acceptable salt or optical isomer thereof.

2. Formula I or Formula II containing a structure represented by the following general formula: 【Chemistry 2】 wherein X is C or N; R １ is selected from H, a substituted or unsubstituted C1-10 alkyl group, and a substituted or unsubstituted C3-C10 cycloalkyl group, wherein the substituent is selected from a halogen, an amino group, a cyano group, a hydroxy group, an aldehyde group, a carboxyl group, and a sulfone group; Y is a cyclic group, and is located at any position of the aromatic ring to which it is bonded, and may be a C3-10 cycloalkyl group, a C6-12 aromatic ring, or a C5-12 heterocycle; the C6-12 aromatic ring includes a C6-12 aromatic ring and a C6-12 aromatic heterocycle, the C6-12 aromatic heterocycle contains at least one heteroatom, the C5-12 heterocycle is a saturated heterocycle or an unsaturated heterocycle, the heterocycle contains 1-3 heteroatoms, and the heteroatom may be O, N or S; R ５ represents H, halogen, an amino group, a cyano group, a C1-10 alkyl group, a C1-10 alkoxy group, a C3-10 cycloalkyl group, -S(O) ２ R ２ , -C(O)R ２ , -NR ３ R ４ , -SR ６ , -OR ６ wherein R ２ is selected from H, an amino group, a halogen atom, a C alkyl group, a C alkoxy group, and a C cycloalkyl group, and said R ３ , R ４ form a 4-8 membered ring containing at least one N together with the N to which they are attached, and ６ is selected from C1-6 alkyl groups; The halogen may be mono- or polysubstituted and is selected from F, Cl, Br, I, In formula II, R １ , R ２ are each independently selected from H, a halogen, a cyano group, an amino group, a substituted or unsubstituted C1-6 alkyl group, and a substituted or unsubstituted C1-6 alkoxy group, wherein the substituted C1-6 alkyl group or C1-6 alkoxy group is further substituted with at least one halogen or hydroxy group; and t in formula II is a chemical bond; 【Transformation 3】 may be one or more selected from the following] or a pharmaceutically acceptable salt or optical isomer thereof.

3. In formula I, X is C or N; R １ is selected from a C1-10 alkyl group and a C3-C10 cycloalkyl group; Y is a cyclic group, and may be a C3-10 cycloalkyl group, a C6-12 aromatic ring, or a C5-12 heterocycle; the C6-12 aromatic ring includes a C6-12 aromatic ring and a C6-12 aromatic heterocycle, the C6-12 aromatic heterocycle contains at least one heteroatom, the C5-12 heterocycle is a saturated heterocycle or an unsaturated heterocycle, the heterocycle contains 1-3 heteroatoms, and the heteroatom may be O, N or S; R ５ represents H, halogen, an amino group, a cyano group, a C1-10 alkyl group, a C1-10 alkoxy group, a C3-10 cycloalkyl group, -S(O) ２ R ２ , -C(O)R ２ , -NR ３ R ４ , -SR ６ , -OR ６ wherein R ２ is selected from H, an amino group, a halogen atom, a C alkyl group, a C alkoxy group, and a C cycloalkyl group, and said R ３ , R ４ form a 4-8 membered ring containing at least one N together with the N to which they are attached, and ６ is selected from C1-6 alkyl groups; The halogen may be mono- or polysubstituted and is selected from F, Cl, Br, I, In formula I, R １ , R ２ are each independently selected from H, halogen, a substituted or unsubstituted C1-3 alkyl group, and a substituted or unsubstituted C1-3 alkoxy group, and the substituted C1-3 alkyl group or C1-3 alkoxy group is further substituted with at least one F.

3. The compound according to claim 2, or a pharmaceutically acceptable salt or optical isomer thereof.

4. In formula I, Y is 【Chemistry 4】 phenyl, pyridyl, thienyl, and thiazolyl groups, and in formula II, R １ , R ２ are each independently selected from H, F, Cl, Br, a methyl group, a methoxy group, and a trifluoromethoxy group; and / or The compound according to claim 2, characterized in that in formula I, R 5 is selected from H, F, Cl, a cyano group, a methyl group, a methylthio group, a methoxy group, a methylsulfonyl group, a methylcarbonyl group, and a methylpiperazine group, and in formula II, R 1 and R 2 are each independently selected from H and F, or a pharmaceutically acceptable salt or optical isomer thereof.

5. in particular, 1) 2-[(2,2-dimethylpropyl){[4-(4-methylpiperazin-1-yl)phenyl]methyl}amino]pyrimidine-4-carbonitrile, 2) 2-[(2,2-dimethylpropyl)[(4-phenylphenyl)methyl]amino]pyrimidine-4-carbonitrile, 3) 2-[(2,2-dimethylpropyl)({4-[4-(methylthio)phenyl]phenyl}methyl)amino]pyrimidine-4-carbonitrile, 4) 2-[(2,2-dimethylpropyl){[4-(4-fluorophenyl)phenyl]methyl}amino]pyrimidine-4-carbonitrile, 5) 2-[(2,2-dimethylpropyl){[4-(4-methoxyphenyl)phenyl]methyl}amino]pyrimidine-4-carbonitrile, 6) 2-[(2,2-dimethylpropyl)({4-[4-(methylsulfonyl)phenyl]phenyl}methyl)amino]pyrimidine-4-carbonitrile, 7) 2-({[4-(5-cyanothiophen-2-yl)phenyl]methyl}(2,2-dimethylpropyl)amino)pyrimidine-4-carbonitrile, 8) 2-({[4-(6-chloropyridin-3-yl)phenyl]methyl}(2,2-dimethylpropyl)amino)pyrimidine-4-carbonitrile, 9) 2-({[4-(3,4-dichlorophenyl)phenyl]methyl}(2,2-dimethylpropyl)amino)pyrimidine-4-carbonitrile, 10) 2-[(2,2-dimethylpropyl){[4-(5-methylthiophen-2-yl)phenyl]methyl}amino]pyrimidine-4-carbonitrile, 11) methyl 4-(4-{[(4-cyanopyrimidin-2-yl)(2,2-dimethylpropyl)amino]methyl}phenyl)benzoate; 12) 2-({[4-(4-chloro-3-fluorophenyl)phenyl]methyl}(2,2-dimethylpropyl)amino)pyrimidine-4-carbonitrile, 13) 2-[(2,2-dimethylpropyl)({4-[4-(4-methylpiperazin-1-yl)phenyl]phenyl}methyl)amino]pyrimidine-4-carbonitrile, 14) 2-[(cyclohexylmethyl)[(4-phenylphenyl)methyl]amino]pyrimidine-4-carbonitrile, 15) 2-[(cyclohexylmethyl)({4-[4-(4-methylpiperazin-1-yl)phenyl]phenyl}methyl)amino]pyrimidine-4-carbonitrile, 16) 2-[(cyclohexylmethyl)({4-[4-(methylsulfonyl)phenyl]phenyl}methyl)amino]pyrimidine-4-carbonitrile, 17) 2-(neopentyl(4-(thiazol-4-yl)benzyl)amino)pyrimidine-4-carbonitrile, 18) 2-((4-(2-(4-methylpiperazin-1-yl)thiazol-4-yl)benzyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 19) (1R,2R)-2-(8-fluoro-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-2-carbonyl)-N-(4-oxotetrahydrofuran-3-yl)cyclohexane-1-formamide; 20) (1R,2R)-2-(2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 21) (1R,2R)-2-(8-fluoro-2,3,4,9-tetrahydro-1H-pyrido-[3,4-b]indole-2-carbonyl)-N-((S)-4-oxotetrahydrofuran-3-yl)cyclohexane-1-formamide; 22) (1R,2R)-2-(6,8-difluoro-2,3,4,9-tetrahydro-1H-pyrido-[3,4-b]indole-2-carbonyl)-N-((S)-4-oxotetrahydrofuran-3-yl)cyclohexane-1-formamide; 23) (1R,2R)-2-(8-chloro-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-((S)-4-oxotetrahydrofuran-3-yl)cyclohexane-1-formamide; 24) (1R,2R)-2-(8-bromo-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-((S)-4-oxotetrahydrofuran-3-yl)cyclohexane-1-formamide; 25) (1R,2R)-2-(6-(trifluoromethoxy)-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 26) (1R,2R)-2-(6-(trifluoromethoxy)-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(S)-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 27) (1R,2R)-2-(6-fluoro-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(S)-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 28) (1R,2R)-2-(6-fluoro-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(S)-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 29) (1R,2R)-2-(6-methoxy-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(S)-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 30) (1R,2R)-2-(6-methoxy-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 31) (1R,2R)-2-(8-fluoro-6-methoxy-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(S)-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 32) (1R,2R)-2-(8-fluoro-6-methoxy-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-2-carbonyl)-N-(4-oxotetrahydrofuran-3-yl)-cyclohexane-1-formamide; 33) 2-((4-chlorobenzyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 34) 2-((4-bromobenzyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 35) 2-((naphthalen-2-ylmethyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 36) 2-(neopentyl(quinolin-2-ylmethyl)amino)pyrimidine-4-carbonitrile, 37) 2-(((4'-(4-(2-hydroxyethyl)piperazin-1-yl)-[1,1'-biphenyl]-4-yl)methyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 38) 2-(((4'-(4-hydroxypiperidin-1-yl)-[1,1'-biphenyl]-4-yl)methyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 39) 2-(((2-methyl-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)methyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 40) 2-(((2-chloro-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)methyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 41) 2-(neopentyl(4-(4-(2-oxopropyl)piperazin-1-yl)benzyl)amino)pyrimidine-4-carbonitrile, 42) 2-(isobutyl((4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)methyl)amino)pyrimidine-4-carbonitrile, 43) N-(4-((4-cyanopyrimidin-2-yl)(neopentyl)amino)methyl)phenyl)-2-(4-methylpiperazin-1-yl)acetamide; 44) 2-((4-(4-methylpiperazine-1-carbonyl)benzyl)(neopentyl)amino)pyrimidine-4-carbonitrile, 45) A compound selected from the group consisting of 4-(((4-cyanopyrimidin-2-yl)(neopentyl)amino)methyl)-N-(2-(4-methylpiperazin-1-yl)-2-oxoethyl)benzamide, or a pharmaceutically acceptable salt or optical isomer thereof.

6. A pharmaceutical composition comprising the compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt or optical isomer thereof.

7. A pharmaceutical composition described in claim 6 for treating a disease targeting cathepsin K.

8. The pharmaceutical composition according to claim 7, wherein the diseases targeting cathepsin K include tumors, thyroid diseases, cardiovascular diseases, bone diseases and gum diseases, preferably the thyroid diseases include hyperthyroidism, preferably the cardiovascular diseases include atherosclerosis, myocardial hypertrophy and heart failure, preferably the bone diseases include osteoporosis, osteoarthritis and rheumatoid arthritis, preferably the gum diseases include gingivitis and periodontitis, and preferably the disease targeting cathepsin K is osteoporosis.

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