Tripterine derivative with osteoporosis treatment activity as well as preparation method and application of tripterine derivative

Abstract

The invention discloses a tripterine derivative with osteoporosis treatment activity and a preparation method and application thereof, and belongs to the technical field of medicines, and the tripterine derivative is any one of general formulas I-VIII shown in the specification. According to the tripterine derivative and the preparation method thereof provided by the invention, aromatization modification is carried out on an A ring of tripterine, and different types of heterocyclic rings are introduced to the left side of the A ring, so that not only is the structural diversity of the tripterine enriched, but also the structure-function relationship is perfected by carrying out structural derivatization on the C-29 site. On the basis of retaining the osteoporosis treatment activity of the tripterine, the cytotoxicity of the tripterine is remarkably reduced, the practical application value of the tripterine serving as an osteoporosis treatment agent is effectively improved, and the tripterine derivative can be applied to treatment of osteoporosis.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a tripterygium wilfordii derivative having activity in treating osteoporosis, and a preparation method and application thereof. Background Art

[0002] Osteoporosis is a systemic bone disease caused by various factors. Its main characteristics are decreased bone density and quality, damage to bone microstructure, and increased bone brittleness, which can lead to fractures. Osteoporosis is mainly divided into primary and secondary types. Primary osteoporosis can be further divided into postmenopausal osteoporosis (type I), senile osteoporosis (type II), and idiopathic osteoporosis. Currently, drug therapy remains the main treatment for osteoporosis.

[0003] Celastrol (tripterine) is a pentacyclic triterpenoid compound with a quinone methyl structure isolated from the root of Tripterygium wilfordii. It has a wide range of biological activities, including anti-osteoporosis, antioxidant, anti-cancer, neuroprotective, anti-viral, anti-obesity, and anti-Alzheimer's disease. The C2 position of the A ring and the C6 position of the B ring of tripterine are highly sensitive to nucleophilic addition reactions. Its quinone methyl structure can undergo Michael addition reactions with the sulfhydryl group of protein cysteine ​​to form Michael addition complexes, which may be the main mechanism for tripterine to exert its activity and produce toxicity. At present, domestic research mainly focuses on the structural modification of the 29-position carboxyl group of tripterine, but these modifications have not effectively improved the problem of tripterine's strong side effects. The present invention designed and synthesized a series of tripterine derivatives. After cell-based activity evaluation, it was proved that these compounds all have good anti-osteoporosis activity. Summary of the Invention

[0004] The present invention aims to provide low-toxic, anti-osteoporosis tripterygium derivatives and their preparation methods. Modification of the A / B rings of tripterygium reduces the cytotoxicity of tripterygium while retaining its anti-osteoporosis activity, thereby enhancing the practical application value of tripterygium as an anti-osteoporosis ingredient.

[0005] The structure of tripterygium wilfordii is:

[0006]

[0007] The technical solutions of the present invention are as follows:

[0008] The tripterygium wilfordii derivative of the present invention is any one of the following general formulas I to VIII:

[0009]

[0010] In general formulas I to VIII, R1 is H, a C1-C6 straight or branched chain alkyl, a cycloalkyl, a 5-12 membered heterocyclic group or a heterocyclic aromatic group, wherein the heterocyclic group or heterocyclic aromatic group contains 1-3 heteroatoms selected from O, S, and N.

[0011] R2 and R3 are independently OH, a halogen atom, a C1-C6 straight or branched alkyl group, a cycloalkyl group, a substituted or unsubstituted 5-12 membered heterocyclic group, a heterocyclic aryl group or a substituted phenyl group, and the substituents are independently C1-C6 straight or branched alkyl group or C1-C6 alkoxy group;

[0012] Alternatively, R2 and R3 together form a cyclic structure, and the formed cyclic structure is a cycloalkyl group, a substituted or unsubstituted phenyl group, a substituted or unsubstituted heterocyclic group or a heterocyclic aromatic group, wherein the substituents are independently OH, a C1-C6 straight-chain or branched alkyl group, a C1-C6 alkoxy group, a halogen atom, a cyano group, or an amino group, and the heterocyclic group or heterocyclic aromatic group contains 1-3 heteroatoms selected from O, S, and N.

[0013] R4 is OH, a halogen atom, a C1-C6 straight or branched alkyl group, a cycloalkyl group, a substituted or unsubstituted 5-12 membered heterocyclic group, a heterocyclic aromatic group or a phenyl group; the substituents are independently C1-C6 straight or branched alkyl group, a C1-C6 alkoxy group, a phenyl group, a cycloalkyl group, a halogen atom, or two adjacent carbon atoms together form a cyclic structure, the cyclic structure may contain 1-3 heteroatoms, and the heteroatoms are selected from O, S, and N.

[0014] R5 and R6 are independently H, C1-C6 straight or branched alkyl, hydroxyl-substituted alkyl, N,N-dimethylamino-substituted alkyl, halogen-substituted alkyl, amino-substituted alkyl, substituted or unsubstituted phenyl, substituted or unsubstituted benzyl, and any one of the following structural formulas; the substituents are independently halogen atoms, C1-C6 straight or branched alkyl, C1-C6 alkoxy, hydroxyl or amino;

[0015]

[0016] Or R5, R6 and the commonly connected N atom form a ring structure, and the formed ring structure includes:

[0017]

[0018] R7 and R8 are independently H, C1-C6 straight or branched alkyl, hydroxyl-substituted alkyl, N,N-dimethylamino-substituted alkyl, halogen-substituted alkyl, amino-substituted alkyl, substituted or unsubstituted phenyl, substituted or unsubstituted benzyl, and any one of the following structural formulas; the substituents are independently halogen atoms, C1-C6 straight or branched alkyl, C1-C6 alkoxy, hydroxyl or amino;

[0019]

[0020] Or R7, R8 and the commonly connected N atom form a ring structure, and the formed ring structure includes:

[0021]

[0022] The halogen atom in the present invention is selected from F, Cl, Br and I.

[0023] Furthermore, the tripterygium wilfordii derivative is any one of the following compounds:

[0024]

[0025]

[0026]

[0027] The present invention also provides a method for preparing the above-mentioned tripterygium wilfordii derivative, comprising the following preparation route:

[0028] Route 1: Preparation of triptolide derivatives represented by general formula I and VII, including compounds 1A, 1A-1, 1-1 to 1-31, 4-1 to 4-5

[0029]

[0030] Using tripterygium wilfordii 1 as the starting material, it reacts with iodomethane to obtain intermediate 2;

[0031] Add methanol and hydrochloric acid to intermediate 2, adjust the pH to 1-2, and react at room temperature to obtain intermediate 3;

[0032] Manganese dioxide is added to intermediate 3 to undergo oxidation reaction to obtain intermediate 4:

[0033] Intermediate 4 reacts with ethylenediamine under reflux to obtain compound 1A;

[0034] Compound 1A is used to prepare compound 1A-1, specifically comprising: (1) adding sodium hydride to a reaction flask, adding hexamethylphosphoramide (HMPA) under nitrogen protection, slowly adding propyl mercaptan dropwise at 0°C, and stirring at room temperature; (2) adding compound 1A to the reaction flask, adding the reaction solution of step (1) under nitrogen protection, and reacting at room temperature to obtain compound 1A-1;

[0035] 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) and N,N-diisopropylethylamine (DIPEA) are added to compound 1A-1 and reacted with the corresponding amine at room temperature to obtain the tripterygium wilfordii derivative shown in general formula VII, or R1OH is added and reacted at room temperature to obtain the tripterygium wilfordii derivative shown in general formula I.

[0036] In route 1, R1, R5 and R6 are as defined above. Further, R1, R5 and R6 are the corresponding groups at the corresponding positions of compounds 1A, 1A-1, 1-1 to 1-31, and 4-1 to 4-5.

[0037] Route 2: Preparation of compounds 2B and 2B-1

[0038]

[0039] Intermediate 3 was prepared by the method of route 1, and then a base was added and reacted with dibromomethane under nitrogen protection to prepare compound 2B; referring to the method of preparing compound 1A-1 from compound 1A in route 1, compound 2B-1 was prepared from compound 2B.

[0040] Route 3: Preparation of compounds 2C and 2C-1

[0041]

[0042] Intermediate 3 was prepared by the method of route 1, and then a base was added and reacted with dibromoethane under nitrogen protection to prepare compound 2C; referring to the method of preparing compound 1A-1 from compound 1A in route 1, compound 2C-1 was prepared from compound 2C.

[0043] Route 4: Preparation of compounds 3A and 3A-1

[0044]

[0045] Intermediate 4 was prepared by the method of route 1, and then reacted with glycine, ammonium acetate, and acetic acid to prepare compound 3A. Referring to the method of preparing compound 1A-1 from compound 1A in route 1, compound 3A was prepared from compound 3A to obtain compound 3A-1.

[0046] Route 5: Preparation of triptolide derivatives represented by formula IX

[0047]

[0048] The intermediate 4 is prepared by the method of route 1, and then reacted with a substituted o-diamino group in anhydrous ethanol to prepare the tripterygium wilfordii derivative shown in the general formula IX.

[0049] In route five, R2 and R3 are defined as described above. Further, R2 and R3 are corresponding groups of compounds 4C-4W at corresponding positions.

[0050] Route six: preparation of compounds of general formula VI, compounds 4C-4W

[0051]

[0052] The intermediate 4 is prepared by the method of route one, and then reacted with a substituted aldehyde compound, ammonium acetate and acetic acid to prepare the triptolide derivative of general formula VI when R1 is methyl. Further, R4 is the corresponding group of compounds 4C-4W at the corresponding position.

[0053] Route seven: preparation of compounds of general formula VI-1, compounds 4F-1

[0054]

[0055] The triptolide derivative of general formula VI is prepared by the method of route six, and then reacted in a system of sodium hydride, hexamethylphosphorus triamide and propanethiol at room temperature under N2 protection to prepare the compound of general formula VI-1. Further, when VI is compound 4F, the prepared compound is 4F-1, and R4 is the corresponding group of compound 4F-1 at the corresponding position.

[0056] Route eight: preparation of compound 2-22

[0057]

[0058] The compound 1A-1 is prepared by the method of route one; the compound 1A-1 is reacted with DPPA to prepare the compound 1A-2; the compound 1A-2 and DIPEA are dissolved in a solvent, 1-Boc-4-(4-piperidyl)-piperazine is added, and then reacted to prepare the intermediate 1A-3, and then deprotected by adding an acid to prepare the compound 2-22.

[0059] A pharmaceutical composition comprising the triptolide derivative or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. The administration mode of the pharmaceutical composition includes injection administration, oral administration, and dosage forms include oral administration dosage forms, injection administration dosage forms, respiratory tract administration dosage forms, and skin administration dosage forms.

[0060] The triptolide derivative of the present application changes both A / B rings of triptolide, retains the anti-osteoporosis activity, and significantly reduces the side effects of triptolide, and thus can be applied to inhibit osteoclast differentiation and treat osteoporosis diseases.

[0061] Therefore, the present invention also provides the use of the tripterine derivative or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising the tripterine derivative in the preparation of a drug for treating osteoporosis.

[0062] Beneficial effects of the present invention:

[0063] The present invention provides a tripterine derivative and a preparation method thereof. By aromatizing the A ring of tripterine and introducing different types of heterocycles to the left of the A ring, the tripterine's structural diversity is enriched. Furthermore, structural derivatization is performed at the C-29 position to improve the structure-activity relationship. While retaining the osteoporosis therapeutic activity of tripterine, the derivative significantly reduces its cytotoxicity, effectively enhancing its practical application as an osteoporosis therapeutic agent and finding application in the treatment of osteoporosis. DETAILED DESCRIPTION

[0064] The technical solutions of the present invention are further described below through examples. Typical compounds of the present invention include the compounds of the present invention or pharmaceutically acceptable salts thereof.

[0065] Example 1: Synthesis of Compounds 1A, 1A-1, and 1-1

[0066] Synthesis of Intermediate 2: In a 250 mL, eggplant-shaped flask, celastrol 1 (10.0 g, 22.0 mmol) and 100 mL of N,N-dimethylformamide were added, and sodium bicarbonate was added with stirring. After 10 minutes, iodomethane (4.1 mL, 0.067 mol) was added. The reaction was stirred at room temperature for 48 hours. After the reaction, the reaction solution was poured into cold water, filtered, and the filter cake was dried under vacuum at 50°C to obtain 9.7 g of red solid intermediate 2, with a yield of 95.4%;

[0067] Synthesis of Intermediate 3: In a 100 mL eggplant-shaped flask, intermediate 2 (4.0 g, 8.62 mmol) was added, followed by 50 mL of methanol. The solid partially dissolved, and 5 mL of hydrochloric acid was added with stirring. The pH of the reaction solution was about 1-2. The reaction was allowed to proceed at room temperature for 12 h. The reaction solution gradually changed from red to light yellow, and a large amount of solid precipitated. The reaction was monitored by TLC. After the reaction was completed, the reaction solution was poured into cold water and filtered. The filter cake was washed twice with n-hexane and dried under reduced pressure at 30°C to obtain light yellow solid intermediate 3 (3.56 g, 7.67 mmol) in a yield of 89%.

[0068] Synthesis of Intermediate 4: To a 100 mL eggplant-shaped flask were added Intermediate 3 (5.0 g, 10.78 mmol), 50 mL of dichloromethane, and manganese dioxide (1.87 g, 21.55 mmol). The mixture was stirred at room temperature. After completion of the reaction, the insoluble matter was removed by filtration. After concentrating the solvent, the residue was recrystallized from ethanol to obtain a red solid Intermediate 4 (4.57 g, 9.43 mmol) in a yield of 87.5%.

[0069] Synthesis of Compound 1A: Intermediate 4 (2.0 g, 4.33 mmol) was added to a 100 mL eggplant-shaped flask. 30 mL of ethanol was added. The solid did not completely dissolve. Ethylenediamine (286 mg, 4.76 mmol) was added and the mixture was refluxed. After approximately 5 h, the starting material disappeared. The reaction solution was returned to room temperature and allowed to stand for precipitation. The product was filtered to obtain Compound 1A (1.48 g, 3.03 mmol) as a white solid in a yield of 70%.

[0070] 1 H NMR(400MHz,Chloroform-d)δ8.73(d,J=1.9Hz,1H),8.70(d,J=1.9Hz,1H),7.82(s,1H) ,6.81(d,J=10.0Hz,1H),6.66(d,J=10.0Hz,1H),5.84(dd,J=6.9,1.8Hz,1H),3.54(s,3H ),2.76(s,3H),2.33–1.96(m,5H),1.93–1.71(m,4H),1.50–1.39(m,2H),1.37–1.29(m,1 H),1.20(s,3H),1.08(s,3H),1.05(s,3H),1.02(s,3H),0.99–0.92(m,1H),0.89(s,3H);

[0071] Synthesis of compound 1A-1: (1) Sodium hydride (4.0 g, 100 mmol) and hexamethylphosphoric acid triamide 50 mL were added to a 100 mL eggplant-shaped flask, stirred for 20 min under N2 protection, and propanethiol (3.8 g, 50 mmol) was slowly added dropwise at 0°C. After the addition was complete, the mixture was reacted at room temperature for 2 h and allowed to stand for use; (2) Compound 1A (2.0 g, 4.13 mmol) was added to a 100 mL eggplant-shaped flask, and 10 mL of the reaction solution of step (1) was added under N2 protection. The mixture was reacted at room temperature for 12 h. After the reaction was completed, water was added at 0°C to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phases were combined, and the organic layer was washed with saturated copper sulfate solution 4 times, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was concentrated. The residue was subjected to silica gel column chromatography to obtain white solid compound 1A-1 (1.26 g, 2.68 mmol) with a yield of 65%.

[0072] 1H NMR(400MHz,Chloroform-d)δ13.62(s,1H),8.75(d,J=1.8Hz,1H),8.72(d,J=1.9Hz,1H),7.73(s ,1H),6.71(d,J=10.0Hz,1H),6.43(d,J=9.9Hz,1H),5.72(d,J=6.4Hz,1H),2.60(s,3H),2.55(d, J=15.9Hz,1H),2.43–2.29(m,2H),2.14(td,J=14.0,4.3Hz,1H),1.98–1.80(m,4H),1.54–1.42(m ,2H),1.37(s,3H),1.31–1.24(m,2H),1.11(s,3H),1.04(s,3H),1.02(s,3H),1.03–0.93(m,4H);

[0073] Synthesis of compound 1-1: Compound 1A-1 (40 mg, 0.085 mmol) and 10 mL of dichloromethane were added to an eggplant-shaped flask, followed by diethylamine (6.2 mg, 0.085 mmol) and N,N-diisopropylethylamine (7 mg, 0.09 mmol) and the mixture was reacted at room temperature. After the reaction was completed, water was added, extracted with dichloromethane, and the mixture was washed with water and saturated brine in sequence. The organic solvents were combined and dried over anhydrous sodium sulfate. The solvent was concentrated and the mixture was purified by silica gel column chromatography to obtain compound 1-1 (35.7 mg, 0.068 mmol) as a white solid in a yield of 80.0%.

[0074] 1 H NMR(400MHz,)δ8.73(d,J=1.9Hz,1H),8.69(d,J=1.9Hz,1H),7.81(s,1H),6.82(d,J=10.0H z,1H),6.64(d,J=10.0Hz,1H),5.81(dd,J=6.8,1.9Hz,1H),3.87–3.56(m,1H),3.40–3.21( m,2H),3.18–3.01(m,1H),2.76(s,3H),2.45–2.32(m,1H),2.27–2.00(m,4H),1.99–1.86(m ,1H),1.83–1.63(m,3H),1.55–1.10(m,14H),1.06(s,3H),1.05(s,3H),1.03–0.92(m,5H).

[0075] Example 2: Synthesis of Compound 1-2

[0076] The synthesis method was the same as that in Example 1, except that diethylamine in the last step was replaced with ethanolamine to obtain a light yellow solid compound 1-2 (26 mg, 0.051 mmol) in a yield of 59.6%.

[0077] 1 H NMR(400MHz,Chloroform-d)δ8.72(d,J=1.9Hz,1H),8.67(d,J=1.9Hz,1H),7.76(s,1H),6 .78(d,J=10.1Hz,1H),6.61(d,J=9.9Hz,1H),6.21(t,J=5.4Hz,1H),5.81(dd,J=6.9,1.8H z,1H),3.70–3.61(m,2H),3.39–3.24(m,2H),3.08–2.81(m,1H),2.73(s,3H),2.29–1.99( m,5H),1.88–1.56(m,6H),1.50–1.29(m,2H),1.21(s,3H),1.10(s,3H),1.05–0.96(m,9H).

[0078] Example 3: Synthesis of Compound 1-3

[0079] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with N,N-dimethylethylenediamine to obtain a light yellow solid compound 1-3 (22 mg, 0.040 mmol) in a yield of 47.5%;

[0080] 1 H NMR(400MHz,Chloroform-d)δ8.72(d,J=1.9Hz,1H),8.69(d,J=1.9Hz,1H),7.80(s,1H),6. 79(d,J=10.1Hz,1H),6.62(d,J=10.0Hz,1H),6.55–6.39(m,1H),5.80(dd,J=6.9,1.8Hz,1H) ,3.26–3.12(m,2H),2.75(s,3H),2.47–2.34(m,2H),2.29–2.00(m,10H),1.93–1.72(m,4H) ,1.60–1.25(m,4H),1.20(s,3H),1.10(s,3H),1.04(s,3H),1.03(s,3H),1.02–0.94(m,4H).

[0081] Example 4: Synthesis of Compound 1-4

[0082] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with piperidine to obtain a light yellow solid compound 1-4 (31 mg, 0.057 mmol) with a yield of 67.1%.

[0083] 1 H NMR(400MHz,Chloroform-d)δ8.73(d,J=1.9Hz,1H),8.70(d,J=1.9Hz,1H),7. 82(s,1H),6.82(d,J=10.0Hz,1H),6.64(d,J=10.0Hz,1H),5.82(dd,J=6.8,1.9 Hz,1H),4.45–2.89(m,4H),2.76(s,3H),2.50–2.19(m,2H),2.17–1.72(m,7H), 1.61–1.26(m,12H),1.11(s,3H),1.05(s,3H),1.05(s,3H),0.99–0.92(m,4H).

[0084] Example 5: Synthesis of Compound 1-5

[0085] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with morpholine to obtain a light yellow solid compound 1-5 (28 mg, 0.052 mmol) in a yield of 61.0%;

[0086] 1 H NMR(400MHz,Chloroform-d)δ8.73(d,J=1.9Hz,1H),8.70(d,J=1.9Hz,1H),7.82(s ,1H),6.80(d,J=10.0Hz,1H),6.65(d,J=10.0Hz,1H),5.83(dd,J=6.7,1.9Hz,1H),3 .99–3.11(m,8H),2.77(s,3H),2.48–2.36(m,1H),2.22–1.92(m,5H),1.86–1.69(m ,3H),1.53–1.28(m,6H),1.11(s,3H),1.05(s,3H),1.05(s,3H),1.00–0.92(m,4H).

[0087] Example 6: Synthesis of Compounds 1-6

[0088] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with piperazine to obtain a light yellow solid compound 1-6 (34 mg, 0.063 mmol) with a yield of 74.0%;

[0089] 1H NMR(400MHz,Chloroform-d)δ8.73(d,J=1.9Hz,1H),8.70(d,J=1.9Hz,1H),7.81(s,1H),6.80(d,J=10.0Hz,1H),6.64(d,J=10.0Hz,1H),5.82(d d,J=6.7,1.9Hz,1H),4.01–3.09(m,4H),2.84–2.74(m,6H),2.42(d,J=1 3.9Hz,1H),2.25–1.71(m,10H),1.51–1.24(m,6H),1.15–0.90(m,13H).

[0090] Example 7: Synthesis of Compound 1-7

[0091] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with N-methylpiperazine to obtain a light yellow solid compound 1-7 (21 mg, 0.039 mmol) in a yield of 45.4%.

[0092] 1 H NMR (400MHz, Chloroform-d) δ8.73(d,J=1.9Hz,1H),8.70(d,J=1.9Hz,1H),7.80(s,1H),6.80(d,J=10.1Hz,1H),6.64(d,J=10.0Hz,1H),5.80(dd ,J=6.8,1.9Hz,1H),4.12–2.99(m,4H),2.76(s,3H),2.45–1.69(m,16H) ,1.54–1.29(m,6H),1.11(s,3H),1.05(s,3H),1.04(s,3H),0.94(m,4H).

[0093] Example 8: Synthesis of Compound 1-8

[0094] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with benzylamine to obtain a light yellow solid compound 1-8 (12 mg, 0.02 mmol) in a yield of 26.1%.

[0095] 1H NMR(400MHz,Chloroform-d)δ8.73(d,J=1.8Hz,1H),8.70(d,J=1.9Hz,1H),7.81(s,1H),7.26–7. 14(m,5H),6.78(d,J=10.1Hz,1H),6.66(d,J=10.0Hz,1H),5.91(t,J=5.4Hz,1H),5.81(dd,J=6.8 ,1.8Hz,1H),4.38–4.24(m,2H),2.78(s,3H),2.21–2.01(m,5H),1.91–1.72(m,4H),1.64–1.52(m ,1H),1.49–1.29(m,2H),1.22(s,3H),1.09(s,3H),1.05(s,3H),1.02(s,3H),1.00–0.91(m,4H).

[0096] Example 9: Synthesis of Compound 1-9

[0097] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with 4-methylbenzylamine to obtain a light yellow solid compound 1-9 (38 mg, 0.067 mmol) in a yield of 78.9%.

[0098] 1 H NMR(400MHz,Chloroform-d)δ8.73(d,J=1.9Hz,1H),8.69(d,J=1.9Hz,1H),7.81(s,1H),7.18–6.94(m,4 H),6.79(d,J=10.1Hz,1H),6.66(d,J=10.0Hz,1H),5.88(t,J=5.3Hz,1H),5.80(dd,J=6.9,1.8Hz,1H),4. 35–4.19(m,2H),2.77(s,3H),2.23(s,3H),2.21–2.01(m,5H),1.89–1.72(m,4H),1.62–1.51(m,1H),1.49 –1.42(m,1H),1.37–1.29(m,1H),1.22(s,3H),1.09(s,3H),1.05(s,3H),1.02(s,3H),0.99–0.92(m,4H).

[0099] Example 10: Synthesis of Compound 1-10

[0100] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with p-fluorobenzylamine to obtain a light yellow solid compound 1-10 (40 mg, 0.069 mmol) in a yield of 80.8%.

[0101] 1 H NMR(400MHz,)δ8.73(d,J=1.9Hz,1H),8.70(d,J=1.8Hz,1H),7.81(s,1H),7.22–7.12(m,2H),6.96–6.86(m,2H), 6.74(d,J=10.1Hz,1H),6.66(d,J=10.0Hz,1H),5.97(t,J=5.6Hz,1H),5.80(dd,J=6.8,1.8Hz,1H),4.35(dd,J=14 .5,5.8Hz,1H),4.20(dd,J=14.5,5.2Hz,1H),2.78(s,3H),2.25–1.95(m,5H),1.89–1.70(m,4H),1.65–1.55(m,1 H),1.49–1.41(m,1H),1.37–1.30(m,1H),1.21(s,3H),1.09(s,3H),1.04(s,3H),1.02(s,3H),0.99–0.82(m,4H).

[0102] Example 11: Synthesis of Compound 1-11

[0103] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with p-chlorobenzylamine to obtain a light yellow solid compound 1-11 (39 mg, 0.066 mmol) in a yield of 78.1%.

[0104] 1 H NMR(400MHz,Chloroform-d)δ8.73(d,J=1.9Hz,1H),8.69(d,J=1.9Hz,1H),7.81(s,1H),7.21(d,J=8.4Hz,2H),7. 13(d,J=8.4Hz,2H),6.75(d,J=10.1Hz,1H),6.68(d,J=10.0Hz,1H),6.01(t,J=5.7Hz,1H),5.80(dd,J=6.9,1.8Hz ,1H),4.35(dd,J=14.6,5.9Hz,1H),4.20(dd,J=14.7,5.2Hz,1H),2.78(s,3H),2.20–1.95(m,5H),1.91–1.74(m,4 H),1.59(m,1H),1.45(m,1H),1.32(m,1H),1.22(s,3H),1.09(s,3H),1.04(s,3H),1.02(s,3H),0.99–0.88(m,4H).

[0105] Example 12: Synthesis of Compounds 1-12

[0106] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with p-bromobenzylamine to obtain a light yellow solid compound 1-12 (26 mg, 0.041 mmol) in a yield of 48.3%;

[0107] 1 H NMR(400MHz,Chloroform-d)δ8.73(d,J=1.9Hz,1H),8.69(d,J=1.9Hz,1H),7.81(s,1H),7.36(d,J=8.3Hz,2H),7. 08(d,J=8.4Hz,2H),6.75(d,J=10.1Hz,1H),6.68(d,J=10.0Hz,1H),6.03(t,J=5.7Hz,1H),5.80(dd,J=6.9,1.8Hz ,1H),4.33(dd,J=14.7,5.9Hz,1H),4.18(dd,J=14.7,5.2Hz,1H),2.78(s,3H),2.22–1.96(m,5H),1.89–1.72(m,4 H),1.59(m,1H),1.45(m,1H),1.32(m,1H),1.22(s,3H),1.09(s,3H),1.04(s,3H),1.02(s,3H),0.99–0.88(m,4H).

[0108] Example 13: Synthesis of Compounds 1-13

[0109] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with 4-methoxybenzylamine to obtain a light yellow solid compound 1-13 (15 mg, 0.025 mmol) in a yield of 29.7%.

[0110] 1H NMR(400MHz,Chloroform-d)δ8.73(d,J=1.9Hz,1H),8.70(d,J=1.9Hz,1H),7.81(s,1H),7.13(d,J=8.6Hz ,2H),6.84–6.72(m,3H),6.66(d,J=10.0Hz,1H),5.86(t,J=5.3Hz,1H),5.80(dd,J=6.9,1.8Hz,1H),4.28 (dd,J=14.3,5.3Hz,1H),4.19(dd,J=14.3,5.1Hz,1H),3.68(s,3H),2.78(s,3H),2.19–2.00(m,5H),1.89 –1.73(m,4H),1.59(m,1H),1.45(m,1H),1.21(m,4H),1.09(s,3H),1.04(s,3H),1.02(s,3H),0.97(m,4H).

[0111] Example 14: Synthesis of Compounds 1-14

[0112] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with 4-methylaminopyridine to obtain a light yellow solid compound 1-14 (31 mg, 0.55 mmol) in a yield of 64.9%;

[0113] 1 H NMR(400MHz,)δ8.73(d,J=1.9Hz,1H),8.70(d,J=1.8Hz,1H),8.49(d,J=5.9Hz,1H),7.13(d,J=6.0Hz,1H),6.7 8(d,J=10.0Hz,1H),6.67(d,J=10.0Hz,1H),6.20(t,J=5.8Hz,1H),5.81(dd,J=6.9,1.8Hz,1H),4.41(dd,J=15 .7,6.1Hz,1H),4.26(dd,J=15.7,5.4Hz,1H),2.77(s,3H),2.23–2.02(m,5H),1.92–1.75(m,4H),1.67–1.59(m ,1H),1.54–1.44(m,1H),1.34(m,1H),1.26(s,3H),1.11(s,3H),1.05(s,3H),1.03(s,3H),1.02–0.94(m,4H).

[0114] Example 15: Synthesis of Compound 1-15

[0115] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with 4-aminopiperidine to obtain a light yellow solid compound 1-15 (23 mg, 0.042 mmol) in a yield of 48.9%.

[0116] 1 H NMR (400MHz, DMSO-d6) δ8.84(s,1H),8.81(s,1H),7.75(s,1H),6.86(d,J=10.0Hz,1H),6.70(d,J=9.9Hz,1H),5.85(d,J=6.4Hz,1H),4.38–3.56 (m,4H),2.99–2.65(m,5H),2.31–1.85(m,7H),1.80–1.62(m,5H),1.47– 1.26(m,3H),1.21(s,3H),1.08(m,5H),0.98(s,6H),0.89–0.70(m,4H).

[0117] Example 16: Synthesis of Compound 1-16

[0118] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with 4-aminopiperidine to obtain a light yellow solid compound 1-16 (28 mg, 0.051 mmol) in a yield of 60.4%;

[0119] 1 H NMR (400MHz, DMSO-d6) δ8.84(d,J=2.1Hz,1H),8.81(d,J=1.8Hz,1H),7.76(s,1H),7.50–7.32 (m,1H),6.86(d,J=10.0Hz,1H),6.68(d,J=9.8Hz,1H),5.85(d,J=6.4Hz,1H),3.71–3.45(m,2H ),3.13–2.90(m,2H),2.75–2.60(m,4H),2.29–2.11(m,3H),2.06–1.93(m,2H),1.88–1.63(m,6 H),1.54–1.20(m,6H),1.10(s,3H),1.05(s,3H),0.98(s,3H),0.97(s,3H),0.93–0.80(m,4H).

[0120] Example 17: Synthesis of Compound 1-17

[0121] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with cyclohexylmethylamine to obtain a light yellow solid compound 1-17 (17 mg, 0.031 mmol) with a yield of 36.3%;

[0122] 1 H NMR(400MHz,Chloroform-d)δ8.72(d,J=1.9Hz,1H),8.69(d,J=1.8Hz,1H),7.80(s,1H),6.79 (d,J=10.1Hz,1H),6.63(d,J=10.0Hz,1H),5.81(dd,J=6.9,1.8Hz,1H),5.75(t,J=5.7Hz,1H), 3.10–3.00(m,1H),2.93–2.82(m,1H),2.75(s,3H),2.26–2.02(m,5H),1.89–1.58(m,10H),1. 47–1.16(m,9H),1.10(s,3H),1.04(s,3H),1.03(s,3H),1.01–0.96(m,4H),0.91–0.80(m,2H).

[0123] Example 18: Synthesis of Compound 1-18

[0124] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with N-hydroxyethylpiperazine to obtain a light yellow solid compound 1-18 (35 mg, 0.060 mmol) in a yield of 71.0%;

[0125] 1 H NMR(400MHz,Chloroform-d)δ8.73(d,J=1.9Hz,1H),8.70(d,J=1.9Hz,1H),7. 81(s,1H),6.80(d,J=10.1Hz,1H),6.65(d,J=10.0Hz,1H),5.82(dd,J=6.7,1.9 Hz,1H),4.15–3.01(m,6H),2.76(s,3H),2.59–1.90(m,12H),1.87–1.68(m,3H) ,1.53–1.24(m,7H),1.11(s,3H),1.05(s,3H),1.05(s,3H),0.98–0.86(m,4H).

[0126] Example 19: Synthesis of Compound 1-19

[0127] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with dimethylaminoethylpiperazine to obtain a light yellow solid compound 1-19 (15 mg, 0.025 mmol) in a yield of 29.1%.

[0128] 1H NMR(400MHz,Chloroform-d)δ8.73(d,J=1.9Hz,1H),8.70(d,J=1.8Hz,1H),7.8 0(s,1H),6.80(d,J=10.1Hz,1H),6.64(d,J=9.9Hz,1H),5.80(dd,J=6.7,1.9Hz, 1H),4.00–2.99(m,4H),2.76(s,3H),2.55–2.32(m,8H),2.26–1.88(m,12H),1.8 6–1.69(m,3H),1.52–1.28(m,6H),1.11(s,3H),1.04(s,6H),1.01–0.86(m,4H).

[0129] Example 20: Synthesis of Compound 1-20

[0130] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with 4-morpholinyl-2-ethylpiperazine to obtain a light yellow solid compound 1-20 (20 mg, 0.030 mmol) in a yield of 35.7%.

[0131] 1 H NMR(400MHz,Chloroform-d)δ8.74(d,J=1.8Hz,1H),8.70(d,J=1.8Hz,1H),7. 81(s,1H),6.80(d,J=10.0Hz,1H),6.64(d,J=10.0Hz,1H),5.80(dd,J=6.7,1.7 Hz,1H),4.22–2.92(m,8H),2.76(s,3H),2.42(m,12H),2.18–1.93(m,5H),1.84 –1.64(m,3H),1.51–1.25(m,7H),1.11(s,3H),1.04(s,6H),1.00–0.87(m,4H).

[0132] Example 21: Synthesis of Compound 1-21

[0133] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with 1-(1-methylpiperidin-4-yl)piperazine to obtain a light yellow solid compound 1-21 (23 mg, 0.036 mmol) in a yield of 42.3%;

[0134] 1H NMR (400 MHz, Chloroform-d) δ 8.73 (d, J = 1.9 Hz, 1H), 8.70 (d, J = 1.9 Hz, 1H), 7.81 (s, 1H), 6.79 (d, J = 10.1 Hz, 1H), 6.63 (d, J = 10.0 Hz, 1H), 5.81 (dd, J = 6.8, 1.8 Hz, 1H), 4.08 - 2.78 (m, 6H), 2.76 (s, 3H), 2.42 (m, 4H), 2.25 - 2.02 (m, 7H), 1.95 - 1.67 (m, 8H), 1.58 - 1.22 (m, 10H), 1.11 (s, 3H), 1.04 (s, 6H), 0.92 (s, 4H).

[0135] Example 22: Synthesis of compound 1-22

[0136] The synthesis was carried out as in example 1, except that diethylamine was replaced by 4-piperazepane piperidine in the last step to give compound 1-22 (33 mg, 0.053 mmol) as a light yellow solid in 62.4% yield;

[0137] 13 C NMR (101 MHz, CDC13) δ 176.1, 144.6, 143.6, 143.3, 142.8, 142.5, 142.3, 141.4, 133.5, 130.6, 125.1, 120.8, 119.9, 62.2, 49.6, 45.9, 45.8, 44.1, 40.8, 40.4, 39.5, 38.7, 36.8, 34.2, 34.0, 34.0, 31.3, 31.3, 30.7, 27.9, 27.8, 23.9, 23.1, 20.2, 19.6, 12.2.

[0138] Example 23: Synthesis of compound 1-23

[0139] The synthesis was carried out as in example 1, except that diethylamine was replaced by N-aminoethylpiperazine in the last step to give compound 1-23 (30 mg, 0.052 mmol) as a light yellow solid in 60.6% yield;

[0140] 1H NMR(400MHz,Chloroform-d)δ8.72(d,J=1.9Hz,1H),8.70(d,J=1.9Hz,1H),7.80(s,1H),6.75( d,J=10.0Hz,1H),6.62(d,J=10.0Hz,1H),6.53–6.39(m,1H),5.79(dd,J=6.8,1.8Hz,1H),3.34 –3.06(m,2H),2.93(m,4H),2.75(s,3H),2.63–2.32(m,7H),2.25–1.97(m,5H),1.92–1.70(m,4 H),1.66–1.30(m,3H),1.19(s,3H),1.11(s,3H),1.04(s,3H),1.03(s,3H),1.00–0.89(m,4H).

[0141] Example 24: Synthesis of Compound 1-24

[0142] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with 4-methyl-1-piperazineethylamine to obtain a light yellow solid compound 1-24 (41 mg, 0.069 mmol) in a yield of 81.4%;

[0143] 1 H NMR(400MHz,Chloroform-d)δ8.73(d,J=1.9Hz,1H),8.69(d,J=1.9Hz,1H),7.80(s,1H),6 .76(d,J=10.1Hz,1H),6.62(d,J=10.0Hz,1H),6.58–6.40(m,1H),5.79(dd,J=6.7,1.8Hz, 1H),3.33–3.15(m,2H),2.75(s,3H),2.64–2.41(m,8H),2.29(s,3H),2.25–1.65(m,10H), 1.64–1.21(m,4H),1.19(s,3H),1.10(s,3H),1.05(s,3H),1.03(s,3H),1.02–0.95(m,4H).

[0144] Example 25: Synthesis of Compound 1-25

[0145] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with N-aminoethylpiperazine to obtain a light yellow solid compound 1-25 (13 mg, 0.021 mmol) in a yield of 25.6%.

[0146] 1H NMR(400MHz,Chloroform-d)δ8.69(d,J=1.8Hz,1H),8.66(d,J=1.8Hz,1H),7.79( s,1H),6.77(d,J=10.1Hz,1H),6.59(d,J=10.0Hz,1H),5.82(dd,J=6.4Hz,1.8Hz,1 H),4.25–2.74(m,8H),2.71(s,3H),2.56–2.22(m,6H),2.21–1.86(m,6H),1.82–1. 65(m,3H),1.51–1.29(m,6H),1.11(s,3H),1.04(s,6H),1.03(s,3H),0.90(m,4H).

[0147] Example 26: Synthesis of Compound 1-26

[0148] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with 2,2,2-trifluoroethylamine to obtain a light yellow solid compound 1-26 (33 mg, 0.061 mmol) in a yield of 71.4%;

[0149] 1 H NMR(400MHz,)δ8.73(d,J=1.9Hz,1H),8.70(d,J=1.8Hz,1H),7.80(s,1H),6.79(d, J=10.0Hz,1H),6.65(d,J=9.9Hz,1H),5.93(t,J=6.2Hz,1H),5.81(dd,J=6.9,1.8Hz ,1H),3.98–3.69(m,2H),2.76(s,3H),2.27–1.99(m,5H),1.92–1.75(m,4H),1.61–1 .35(m,3H),1.22(s,3H),1.11(s,3H),1.05(s,3H),1.03(s,3H),0.99–0.89(m,4H).

[0150] Example 27: Synthesis of Compound 1-27

[0151] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with isobutylamine to obtain a light yellow solid compound 1-27 (16 mg, 0.030 mmol) in a yield of 35.6%.

[0152] 1H NMR(400MHz,Chloroform-d)δ8.72(d,J=1.9Hz,1H),8.69(d,J=1.9Hz,1H),7.80(s,1H),6.79 (d,J=10.1Hz,1H),6.64(d,J=10.0Hz,1H),5.81(dd,J=6.9,1.9Hz,1H),5.72(t,J=5.7Hz,1H), 3.09–2.85(m,2H),2.76(s,3H),2.25–2.01(m,5H),1.88–1.61(m,6H),1.49–1.29(m,2H),1.2 0(s,3H),1.10(s,3H),1.05(s,3H),1.03(s,3H),1.02–0.96(m,4H),0.87(s,3H),0.85(s,3H).

[0153] Example 28: Synthesis of Compound 1-28

[0154] The synthesis method was the same as that in Example 1, except that diethylamine in the last step was replaced with ethylenediamine to obtain a light yellow solid compound 1-28 (22 mg, 0.043 mmol) in a yield of 50.0%;

[0155] 1 H NMR (400MHz, DMSO-d6) δ8.84(d,J=1.9Hz,1H),8.82(d,J=1.9Hz,1H),7.89–7.58( m,2H),6.85(d,J=10.1Hz,1H),6.69(d,J=10.0Hz,1H),5.85(d,J=6.4Hz,1H),3.2 5–2.99(m,2H),2.78–2.62(m,4H),2.24–2.08(m,3H),2.03–1.56(m,4H),1.47–1. 19(s,3H),1.12(s,3H),1.06(s,3H),0.98(s,3H),0.97(s,3H),0.89–0.79(m,5H).

[0156] Example 29: Synthesis of Compound 1-29

[0157] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with 1-(2-pyrimidinyl)piperazine to obtain a light yellow solid compound 1-29 (32 mg, 0.052 mmol) in a yield of 61.6%.

[0158] 1H NMR (400 MHz, Chloroform-d) δ 8.71 (d, J = 1.8 Hz, 1H), 8.67 (d, J = 1.8 Hz, 1H), 8.24 (d, J = 4.7 Hz, 2H), 7.78 (s, 1H), 6.79 (d, J = 10.0 Hz, 1H), 6.63 (d, J = 10.0 Hz, 1H), 6.44 (t, J = 4.7 Hz, 1H), 5.77 (dd, J = 6.7, 1.9 Hz, 1H), 4.65 - 2.96 (m, 8H), 2.74 (s, 3H), 2.45 (d, J = 13.9 Hz, 1H), 2.33 - 1.98 (m, 4H), 1.97 - 1.68 (m, 4H), 1.57 - 1.22 (m, 6H), 1.13 (s, 3H), 1.04 (s, 6H), 1.00 - 0.90 (m, 4H).

[0159] Example 30: Synthesis of compound 1-31

[0160] The synthesis was performed as in Example 1, except that diethylamine was replaced by N-(2-aminoethyl)morpholine in the last step, to give compound 1-31 (29 mg, 0.049 mmol) as a light yellow solid in 58.2% yield;

[0161] 1 H NMR (400 MHz, Chloroform-d) δ 8.72 (d, J = 1.9 Hz, 1H), 8.70 (d, J = 1.9 Hz, 1H), 7.80 (s, 1H), 6.74 (d, J = 10.1 Hz, 1H), 6.61 (d, J = 10.0 Hz, 1H), 6.37 (t, J = 4.5 Hz, 1H), 5.79 (dd, J = 6.8, 1.8 Hz, 1H), 3.71 (td, J = 4.8, 2.6 Hz, 4H), 3.32 - 3.06 (m, 2H), 2.75 (s, 3H), 2.52 - 2.33 (m, 5H), 2.21 - 1.96 (m, 5H), 1.91 - 1.73 (m, 4H), 1.63 (dd, J = 15.5, 4.4 Hz, 1H), 1.48 (dt, J = 14.1, 3.6 Hz, 1H), 1.36 - 1.26 (m, 2H), 1.19 (s, 3H), 1.11 (s, 3H), 1.04 (s, 3H), 1.03 (s, 3H), 1.01 - 0.95 (m, 4H).

[0162] Example 31: Synthesis of compound 4-1

[0163] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with glycine carboxylpiperazinepiperidine (prepared by amide condensation reaction of Boc-glycine with 4-piperazinepiperidine, followed by removal of the Boc protecting group with trifluoroacetic acid) to obtain compound 4-1 as a pale yellow solid (39 mg, 0.058 mmol) in a yield of 67.8%;

[0164] 1 H NMR(400MHz,Chloroform-d)δ8.72(s,1H),8.69(s,1H),7.81(s,1H),7.03–6.89(m,1H),6.86–6 .74(m,1H),6.69–6.54(m,1H),5.82(t,J=5.8Hz,1H),4.52(t,J=13.7Hz,1H),4.00–3.81(m,2H) ,3.69(d,J=13.6Hz,1H),3.01–2.60(m,9H),2.54–2.31(m,5H),2.30–1.99(m,6H),1.94–1.71(m ,6H),1.66–1.29(m,5H),1.21(s,3H),1.10(s,3H),1.04(s,3H),1.02(s,3H),1.00–0.87(m,4H).

[0165] Example 32: Synthesis of Compound 4-2

[0166] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with 3-aminopropionylpiperazinepiperidine (prepared by amide condensation reaction of Boc-beta-alanine and 4-piperazinepiperidine, followed by removal of the Boc protecting group with trifluoroacetic acid) to obtain a pale yellow solid compound 4-2 (41 mg, 0.059 mmol) in a yield of 69.2%.

[0167] 1 H NMR(400MHz,Chloroform-d)δ8.85–8.58(m,2H),7.83(s,1H),6.82(d,J=10.1Hz,1H), 6.60(d,J=9.7Hz,1H),6.49(t,J=6.2Hz,1H),5.81(m,1H),4.48(m,1H),3.80–3.45(m, 2H),3.40–3.19(m,1H),2.94–2.71(m,7H),2.58–1.96(m,14H),1.95–1.66(m,6H),1.5 0–1.23(m,6H),1.16(s,3H),1.07(s,3H),1.04(s,3H),1.01(s,3H),0.96–0.87(m,4H).

[0168] Example 33: Synthesis of Compound 4-3

[0169] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with 4-aminobutyrylpiperazinepiperidine (prepared by amide condensation reaction of N-Boc-γ-aminobutyric acid and 4-piperazinepiperidine, followed by removal of the Boc protecting group using trifluoroacetic acid) to obtain a light yellow solid compound 4-3 (43 mg, 0.060 mmol) in a yield of 70.6%.

[0170] 1 H NMR(400MHz,Chloroform-d)δ8.72(d,J=1.9Hz,1H),8.69(d,J=1.9Hz,1H),6.95–6.73(m,2H) ,6.63(d,J=10.0Hz,1H),5.80(m,1H),4.63(d,J=13.2Hz,1H),3.97–3.73(m,1H),3.14(m,2H), 2.99–2.81(m,5H),2.75(s,3H),2.61–2.41(m,5H),2.36(m,3H),2.28–2.01(m,6H),1.90–1.7 1(m,8H),1.60–1.28(m,5H),1.17(s,3H),1.09(s,3H),1.04(s,3H),1.02(s,3H),0.97(m,4H).

[0171] Example 34: Synthesis of Compound 4-5

[0172] The synthesis method was the same as that in Example 1, except that the diethylamine in the last step was replaced with 1-aminocyclopropane-1-carboxylpiperazinepiperidine (prepared by amide condensation reaction of Boc-1-aminocyclopropylcarboxylic acid with 4-piperazinepiperidine, followed by removal of the Boc protecting group with trifluoroacetic acid) to obtain compound 4-5 as a pale yellow solid (42 mg, 0.060 mmol) in a yield of 70.4%;

[0173] 1H NMR(400MHz,Chloroform-d)δ8.73(d,J=1.9Hz,1H),8.70(d,J=1.9Hz,1H),7.80(s,1H ),6.81(d,J=10.0Hz,1H),6.66(d,J=10.0Hz,1H),6.55(s,1H),5.78(d,J=6.4Hz,1H),2 .76(s,3H),2.71–2.56(m,5H),2.27–2.11(m,7H),2.08–1.69(m,11H),1.66–1.41(m,4 H),1.35–1.11(m,8H),1.11–1.06(m,5H),1.05(s,3H),1.02(s,3H),1.00–0.88(m,4H).

[0174] Example 35: Synthesis of Compounds 2B and 2B-1

[0175] Intermediate 3 was prepared by the method in Example 1; intermediate 3 (1.0 g, 2.10 mmol) was added to an eggplant-shaped flask, potassium carbonate (725 mg, 5.25 mmol) was added, and N,N-dimethylformamide 5 mL and dibromomethane (399 mg, 2.31 mmol) were added under nitrogen protection. The reaction was allowed to react at room temperature and monitored by TLC. After the reaction was completed, water was added, extracted with ethyl acetate, washed with water and saturated brine, dried over anhydrous sodium sulfate, and separated by silica gel column chromatography to obtain compound 2B;

[0176] 1 H NMR(400MHz,Chloroform-d)δ6.68(s,1H),6.42(d,J=10.0Hz,1H),6.20(d,J=9 .9Hz,1H),5.91(d,J=1.4Hz,1H),5.87(d,J=1.4Hz,1H),5.56(dd,J=6.8,1.8Hz, 1H),3.53(s,3H),2.31–1.91(m,8H),1.87–1.63(m,4H),1.59–1.34(m,3H),1.19 (s,3H),1.05(s,3H),1.03(s,3H),0.95(s,3H),0.92–0.88(m,1H),0.87(s,3H).

[0177] Synthesis of compound 2B-1: (1) Sodium hydride (4.0 g, 100 mmol) and hexamethylphosphoric acid triamide 50 mL were added to a 100 mL eggplant-shaped flask, stirred for 20 min under N2 protection, and propanethiol (3.8 g, 50 mmol) was slowly added dropwise at 0°C. After the addition was complete, the mixture was reacted at room temperature for 2 h and allowed to stand for use; (2) Compound 2B (2.0 g, 4.20 mmol) was added to a 100 mL eggplant-shaped flask, and 10 mL of the reaction solution of step (1) was added under N2 protection. The mixture was reacted at room temperature for 12 h. After the reaction was completed, water was added at 0°C to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phases were combined, and the organic layer was washed with saturated copper sulfate solution 4 times, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was concentrated. The residue was subjected to silica gel column chromatography to obtain white solid compound 2B-1 (0.89 g, 1.93 mmol) with a yield of 46%.

[0178] 1 H NMR(400MHz,Chloroform-d)δ6.67(s,1H),6.42(d,J=10.0Hz,1H),6.19(d,J=9.9Hz,1H ),5.91(d,J=1.5Hz,1H),5.88(d,J=1.5Hz,1H),5.55(dd,J=6.9,1.8Hz,1H),2.22(d,J= 14.8Hz,2H),2.16(s,3H),2.09–1.96(m,2H),1.79–1.67(m,3H),1.45–1.37(m,2H),1.3 1–1.26(m,3H),1.22(s,3H),1.05(s,3H),1.02(s,3H),1.01(s,3H),0.96–0.90(m,4H).

[0179] Example 36: Synthesis of Compounds 2C and 2C-1

[0180] Intermediate 3 was prepared by the method in Example 1; intermediate 3 (1.0 g, 2.10 mmol) was added to an eggplant-shaped flask, potassium carbonate (725 mg, 5.25 mmol) was added, and N,N-dimethylformamide 5 mL and 1,2-dibromoethane (399 mg, 2.31 mmol) were added under nitrogen protection. The reaction was allowed to react at room temperature and monitored by TLC. After the reaction was completed, water was added, extracted with ethyl acetate, washed with water and saturated brine, dried over anhydrous sodium sulfate, and separated by silica gel column chromatography to obtain compound 2C;

[0181] 1H NMR(400MHz,Chloroform-d)δ6.69(s,1H),6.42(d,J=10.0Hz,1H),6.26(d,J=9.9Hz,1H),5.54(dd,J=6.7,1.8Hz,1H),4.32–4.18(m,4H),3.52(s, 3H),2.28–1.91(m,8H),1.83–1.65(m,4H),1.49–1.34(m,3H),1.18(s,3H) ),1.05(s,3H),1.02(s,3H),0.95(s,3H),0.91–0.88(m,1H),0.86(s,3H).

[0182] Synthesis of compound 2C-1: (1) Sodium hydride (4.0 g, 100 mmol) and hexamethylphosphoric triamide 50 mL were added to a 100 mL eggplant-shaped flask, stirred for 20 min under N2 protection, and propanethiol (3.8 g, 50 mmol) was slowly added dropwise at 0°C. After the addition was complete, the mixture was reacted at room temperature for 2 h and allowed to stand for use; (2) Compound 2C (2.0 g, 4.08 mmol) was added to a 100 mL eggplant-shaped flask, and 10 mL of the reaction solution of step (1) was added under N2 protection. The mixture was reacted at room temperature for 12 h. After the reaction was completed, water was added at 0°C to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phases were combined, and the organic layer was washed with saturated copper sulfate solution 4 times, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was concentrated. The residue was subjected to silica gel column chromatography to obtain white solid compound 2C-1 (0.932 g, 1.96 mmol) with a yield of 48%.

[0183] 1 H NMR(400MHz,Chloroform-d)δ6.68(s,1H),6.43(d,J=10.0Hz,1H),6.26(d,J=10.0Hz,1H),5.59–5.44(m,1H),4.33–4.15(m,4H),2.22(d,J=15.0Hz,2H ),2.13(s,3H),2.08–1.98(m,2H),1.80–1.69(m,4H),1.45–1.38(m,2H),1. 31–1.21(m,5H),1.05(s,3H),1.02(s,3H),1.00(s,3H),0.96–0.88(m,4H).

[0184] Example 37: Synthesis of Compounds 3A and 3A-1

[0185] Intermediate 4 was prepared according to the procedure described in Example 1. A 100 mL jar was charged with intermediate 4 (1.0 g, 2.10 mmol), ammonium acetate (6.66 g, 86.6 mmol), acetic acid 60 mL, glycine (649 mg, 8.7 mmol), and the reaction was stirred at room temperature. The reaction was monitored by TLC. Upon completion, the reaction was quenched with water. The organic layer was extracted with ethyl acetate, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to afford compound 3A;

[0186] 1 H NMR (400 MHz, Chloroform-d) δ 8.02 (s, 1H), 7.52 (s, 1H), 6.62 (d, J = 10.0 Hz, 1H), 6.41 (d, J = 9.9 Hz, 1H), 5.66 (dd, J = 6.8, 1.8 Hz, 1H), 3.53 (s, 3H), 2.47 (s, 3H), 2.27 - 2.00 (m, 5H), 1.89 - 1.70 (m, 4H), 1.48 - 1.38 (m, 2H), 1.36 - 1.26 (m, 1H), 1.20 (s, 3H), 1.07 (s, 3H), 1.02 (s, 3H), 0.99 (s, 3H), 0.97 - 0.91 (m, 1H), 0.89 (s, 3H).

[0187] Synthesis of compound 3A-1: (1) A 100 mL jar was charged with sodium hydride (4.0 g, 100 mmol), hexamethylphosphoramide 50 mL, and stirred for 20 min under N2protection. Propyl mercaptan (3.8 g, 50 mmol) was added slowly at 0 °C. The reaction was stirred at room temperature for 2 h. The reaction was used directly. (2) A 100 mL jar was charged with compound 3A (2.0 g, 4.23 mmol), and the reaction solution from step (1) 10 mL was added at room temperature. The reaction was stirred for 12 h. Upon completion, the reaction was quenched with water at 0 °C. The organic layer was extracted with ethyl acetate, washed with saturated copper sulfate solution 4 times, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography to afford compound 3A-1 (1.11 g, 2.41 mmol) as a white solid in 57% yield;

[0188] 1H NMR(400MHz,Chloroform-d)δ8.06(s,1H),7.49(s,1H),6.60(d,J=10.0Hz,1H),6.35(d,J=9.9Hz,1H),5.62(dd,J=6.9,1.8Hz,1H),2 .43(s,3H),2.31–1.97(m,5H),1.86–1.69(m,4H),1.47–1.35(m,2H),1.34–1.17(m,4H),1.06(s,3H),1.00(s,6H),0.99–0.89(m,4H).

[0189] Example 38: Synthesis of Compound 1B

[0190] Intermediate 4 (2.0 g, 4.33 mmol) was added to a 100 mL eggplant-shaped flask. 30 mL of ethanol was added. The solid did not completely dissolve. 1,2-diaminocyclohexane (543 mg, 4.76 mmol) was added and the mixture was refluxed. After about 5 h, the starting material disappeared. The reaction solution was returned to room temperature and allowed to stand for precipitation. The product was filtered to obtain compound 1B (1.25 g, 2.33 mmol) as a white solid in a yield of 53.8%.

[0191] 1 H NMR(400MHz,Chloroform-d)δ7.70(s,1H),6.73(d,J=10.0Hz,1H),6.61(d,J= 10.0Hz,1H),5.79(d,J=6.6Hz,1H),3.54(s,3H),3.12(s,4H),2.72(s,3H),2.3 3–1.96(m,9H),1.93–1.71(m,4H),1.51–1.38(m,2H),1.36–1.25(m,1H),1.20( s,3H),1.07(s,3H),1.02(s,3H),1.00(s,3H),0.97–0.92(m,1H),0.89(s,3H).

[0192] Example 39: Synthesis of Compound 1C

[0193] The synthesis method was the same as that of Example 38, except that 1,2-diaminocyclohexane was replaced with o-phenylenediamine to obtain a light yellow solid compound 1C (1.23 g, 2.31 mmol) in a yield of 53.4%.

[0194] 1H NMR (400MHz, Chloroform-d) δ8.25–8.15(m,2H),7.95(s,1H),7.82–7.71(m,2H),6.87(d,J=10.0Hz,1H),6.74(d,J=10.0Hz,1H),5.92(d,J=6.5 Hz,1H),3.55(s,3H),2.90(s,3H),2.29–2.04(m,5H),1.94–1.73(m,4H) ,1.46(m,3H),1.21(s,3H),1.09(s,6H),1.04(s,3H),0.99–0.85(m,4H).

[0195] Example 40: Synthesis of Compound 1D

[0196] The synthesis method was the same as that of Example 38, except that 1,2-diaminocyclohexane was replaced with 4,5-diamino-o-diphenylnitrile to obtain a light yellow solid compound 1D (1.48 g, 2.53 mmol) in a yield of 58.4%.

[0197] 1 H NMR(400MHz,Chloroform-d)δ8.66(s,1H),8.58(s,1H),7.89(s,1H),6.96(d, J=10.0Hz,1H),6.70(d,J=10.0Hz,1H),5.92(dd,J=6.8,1.8Hz,1H),3.48(s,3H ),2.79(s,3H),2.24–1.84(m,6H),1.81–1.65(m,3H),1.43–1.25(m,3H),1.14( s,3H),1.03(s,3H),1.02(s,3H),0.98(s,3H),0.94–0.86(m,1H),0.80(s,3H).

[0198] Example 41: Synthesis of Compound 1E

[0199] The synthesis method was the same as that of Example 38, except that 1,2-diaminocyclohexane was replaced with 4,5-difluoro-o-phenylenediamine to obtain a light yellow solid compound 1E (1.41 g, 2.47 mmol) in a yield of 57.0%.

[0200] 1H NMR (400MHz, CDCl3) δ7.98–7.82(m,3H),6.89(d,J=10.0Hz,1H),6.73(d,J= 10.0Hz,1H),5.91(dd,J=6.9,1.9Hz,1H),3.55(s,3H),2.85(s,3H),2.32–1. 99(m,5H),1.95–1.74(m,4H),1.52–1.40(m,2H),1.37–1.31(m,1H),1.21(s, 3H),1.09(s,3H),1.08(s,3H),1.04(s,3H),0.99–0.93(m,1H),0.89(s,3H).

[0201] Example 42: Synthesis of Compound 1F

[0202] The synthesis method was the same as that in Example 1, except that 1,2-diaminocyclohexane in the last step was replaced with 4,5-dichloro-o-phenylenediamine to obtain a light yellow solid compound 1F (1.57 g, 2.61 mmol) in a yield of 60.2%.

[0203] 1 H NMR(400MHz,Chloroform-d)δ8.56(s,1H),8.49(s,1H),7.88(s,1H),6.91(d,J= 10.0Hz,1H),6.73(d,J=10.0Hz,1H),5.92(dd,J=6.9,1.8Hz,1H),3.54(s,3H),2 .83(s,3H),2.33–1.97(m,5H),1.95–1.71(m,4H),1.51–1.38(m,2H),1.38–1.29 (m,1H),1.21(s,3H),1.08(s,6H),1.04(s,3H),1.00–0.91(m,1H),0.88(s,3H).

[0204] Example 43: Synthesis of Compound 1G

[0205] The synthesis method was the same as that in Example 1, except that 1,2-diaminocyclohexane in the last step was replaced with 4,5-dibromo-o-phenylenediamine to obtain a light yellow solid compound 1G (2.06 mg, 2.99 mmol) in a yield of 69.0%.

[0206] 1H NMR (400MHz, CDCl3) δ8.35(s,1H),8.28(s,1H),7.89(s,1H),6.90(d,J=10.0H z,1H),6.73(d,J=10.0Hz,1H),5.92(dd,J=6.9,1.8Hz,1H),3.54(s,3H),2.84 (s,3H),2.34–1.97(m,5H),1.96–1.68(m,4H),1.52–1.38(m,2H),1.37–1.29( m,1H),1.21(s,3H),1.08(s,6H),1.04(s,3H),1.00–0.92(m,1H),0.88(s,3H).

[0207] Example 44: Synthesis of Compound 1H

[0208] The synthesis method was the same as that in Example 1, except that 1,2-diaminocyclohexane in the last step was replaced with 4,5-dimethyl-o-phenylenediamine to obtain a light yellow solid compound 1H (1.37 g, 2.43 mmol) in a yield of 56.2%.

[0209] 1 H NMR(400MHz,Chloroform-d)δ7.96(s,1H),7.91(s,1H),7.90(s,1H),6.83(d,J=10.0H z,1H),6.72(d,J=10.0Hz,1H),5.89(dd,J=6.9,1.8Hz,1H),3.54(s,3H),2.88(s,3H),2 .54(s,3H),2.53(s,3H),2.34–1.97(m,5H),1.94–1.65(m,4H),1.50–1.38(m,2H),1.3 8–1.26(m,1H),1.20(s,3H),1.08(s,6H),1.03(s,3H),0.99–0.92(m,1H),0.89(s,3H).

[0210] Example 45: Synthesis of Compound 4C

[0211] Intermediate 4 was prepared by the method in Example 1; intermediate 4 (1.0 g, 2.10 mmol) was added to an eggplant-shaped flask, followed by ammonium acetate (6.66 g, 86.6 mmol), 60 mL of acetic acid, and p-tert-butylbenzaldehyde (706 mg, 4.36 mmol). The reaction was carried out at 80° C. and monitored by TLC. After the reaction was completed, water was added, extracted with ethyl acetate, washed with water and saturated brine, dried over anhydrous sodium sulfate, and separated by silica gel column chromatography to obtain compound 4C (768 mg, 1.27 mmol) in a yield of 60.5%.

[0212] 1 H NMR(400MHz,Chloroform-d)δ8.17(d,J=8.4Hz,2H),7.53(d,J=8.3Hz,2H),7.50(s,1H ),6.60(d,J=10.0Hz,1H),6.42(d,J=10.0Hz,1H),5.67(dd,J=7.0,1.8Hz,1H),3.53(s, 3H),2.52(s,3H),2.28–2.00(m,5H),1.85–1.70(m,4H),1.49–1.40(m,3H),1.37(s,9H) ),1.20(s,3H),1.07(s,3H),1.05(s,3H),1.00(s,3H),0.97–0.92(m,1H),0.90(s,3H).

[0213] Example 46: Synthesis of Compound 4D

[0214] The synthesis method was the same as that of Example 45, except that p-tert-butylbenzaldehyde was replaced with p-phenylbenzaldehyde to obtain a pale yellow solid compound 4D (820 mg, 1.31 mmol) in a yield of 62.5%.

[0215] 1 H NMR(400MHz,Chloroform-d)δ8.32(d,J=8.4Hz,2H),7.75(d,J=8.4Hz,2H),7.67(d,J=7.4Hz,2H),7 .53(s,1H),7.48(d,J=7.6Hz,2H),7.43–7.35(m,1H),6.62(d,J=10.0Hz,1H),6.43(d,J=9.9Hz,1H) ,5.68(d,J=6.5Hz,1.8Hz,1H),3.54(s,3H),2.54(s,3H),2.26–1.99(m,5H),1.89–1.70(m,4H),1.4 8–1.38(m,3H),1.20(s,3H),1.08(s,3H),1.05(s,3H),1.01(s,3H),0.97–0.95(m,1H),0.91(s,3H).

[0216] Example 47: Synthesis of Compound 4F

[0217] The synthesis method was the same as that of Example 45, except that p-tert-butylbenzaldehyde was replaced with benzaldehyde to obtain a light yellow solid compound 4F (632 mg, 1.15 mmol) in a yield of 54.9%.

[0218] 1H NMR (400MHz, Chloroform-d) δ8.25 (dd, J=6.7, 3.3Hz, 2H), 7.65–7.38 (m, 4H), 6. 61(d,J=9.9Hz,1H),6.42(d,J=10.1Hz,1H),5.67(dd,J=6.5Hz,1.8Hz,1H),3.53( s,3H),2.52(s,3H),2.27–1.96(m,5H),1.89–1.66(m,4H),1.47–1.27(m,3H),1.2 0(s,3H),1.07(s,3H),1.04(s,3H),1.00(s,3H),0.97–0.94(m,1H),0.91(s,3H).

[0219] Example 48: Synthesis of Compound 4G

[0220] The synthesis method was the same as that of Example 45, except that p-tert-butylbenzaldehyde was replaced with cyclohexylcarboxaldehyde to obtain a light yellow solid compound 4G (647 mg, 1.17 mmol) in a yield of 55.5%;

[0221] 1 H NMR(400MHz,Chloroform-d)δ7.42(s,1H),6.57(d,J=10.0Hz,1H),6.38(d,J=9.9Hz,1H),5.61(dd,J=6.9,1.8Hz,1H),3.52(s,3H),2.93(m,1H),2.43 (s,3H),2.30–1.94(m,8H),1.89–1.63(m,8H),1.48–1.24(m,6H),1.19(s,3 H),1.06(s,3H),1.02(s,3H),0.98(s,3H),0.97–0.91(m,1H),0.89(s,3H).

[0222] Example 49: Synthesis of Compound 4H

[0223] The synthesis method was the same as that of Example 45, except that p-tert-butylbenzaldehyde was replaced with p-methoxybenzaldehyde to obtain a pale yellow solid compound 4H (704 mg, 1.22 mmol) in a yield of 57.9%;

[0224] 1H NMR(400MHz,Chloroform-d)δ8.19(d,J=8.8Hz,1H),7.48(s,1H),7.02(d,J=8.9Hz,1H ),6.59(d,J=10.0Hz,1H),6.41(d,J=9.9Hz,1H),5.66(dd,J=6.9,1.8Hz,1H),3.89(s, 3H),3.53(s,3H),2.51(s,3H),2.40–1.91(m,5H),1.94–1.64(m,4H),1.44–1.25(m,3H ),1.20(s,3H),1.07(s,3H),1.04(s,3H),1.00(s,3H),0.96–0.93(m,1H),0.91(s,3H).

[0225] Example 50: Synthesis of Compound 4I

[0226] The synthesis method was the same as that of Example 50, except that p-tert-butylbenzaldehyde was replaced with 4-pyridinecarboxaldehyde to obtain a light yellow solid compound 4I (635 mg, 1.15 mmol) in a yield of 55.0%;

[0227] 1 H NMR(400MHz,Chloroform-d)δ8.82(d,J=4.9Hz,2H),8.08(d,J=4.9Hz,2H),7.55(s, 1H),6.66(d,J=10.0Hz,1H),6.43(d,J=9.8Hz,1H),5.68(dd,J=6.8Hz,1.8Hz,1H),3 .54(s,3H),2.53(s,3H),2.30–1.96(m,5H),1.91–1.68(m,4H),1.46–1.28(m,3H),1 .20(s,3H),1.07(s,3H),1.04(s,3H),1.00(s,3H),0.97–0.93(m,1H),0.90(s,3H).

[0228] Example 51: Synthesis of Compound 4J

[0229] The synthesis method was the same as that of Example 45, except that p-tert-butylbenzaldehyde was replaced with p-methylbenzaldehyde to obtain a light yellow solid compound 4J (666 mg, 1.18 mmol) in a yield of 56.3%.

[0230] 1H NMR (400 MHz, Chloroform-d) δ 8.13 (d, J = 7.7 Hz, 2H), 7.50 (s, 1H), 7.32 (d, J = 7.7 Hz, 2H), 6.60 (d, J = 10.0 Hz, 1H), 6.42 (d, J = 10.1 Hz, 1H), 5.66 (dd, J = 6.7 Hz, 1.8 Hz, 1H), 3.53 (s, 3H), 2.51 (s, 3H), 2.44 (s, 3H), 2.29 - 1.94 (m, 5H), 1.88 - 1.69 (m, 4H), 1.44 - 1.27 (m, 3H), 1.20 (s, 3H), 1.07 (s, 3H), 1.04 (s, 3H), 1.00 (s, 3H), 0.97 - 0.93 (m, 1H), 0.90 (s, 3H).

[0231] Example 52: Synthesis of compound 4K

[0232] The synthesis was carried out as in Example 45, except that p-tert- butylbenzaldehyde was replaced by p-propylaldehyde, to give compound 4K (527 mg, 1.05 mmol) as a light yellow solid in 50.1% yield;

[0233] 1 H NMR (400 MHz, Chloroform-d) δ 8.13 (d, J = 7.7 Hz, 2H), 7.50 (s, 1H), 7.32 (d, J = 7.7 Hz, 2H), 6.60 (d, J = 10.0 Hz, 1H), 6.42 (d, J = 10.1 Hz, 1H), 5.66 (dd, J = 6.7 Hz, 1.8 Hz, 1H), 3.53 (s, 3H), 2.51 (s, 3H), 2.44 (s, 3H), 2.29 - 1.94 (m, 5H), 1.88 - 1.69 (m, 4H), 1.44 - 1.27 (m, 3H), 1.20 (s, 3H), 1.07 (s, 3H), 1.04 (s, 3H), 1.00 (s, 3H), 0.97 - 0.93 (m, 1H), 0.90 (s, 3H).

[0234] Example 53: Synthesis of compound 4L

[0235] The synthesis was carried out as in Example 45, except that p-tert- butylbenzaldehyde was replaced by p-propylaldehyde, to give compound 4K (527 mg, 1.05 mmol) as a light yellow solid in 50.1% yield;

[0236] 1H NMR(400MHz,Chloroform-d)δ7.41(s,1H),6.57(d,J=10.0Hz,1H),6.38(d,J=9.9Hz,1H),5.63(d,J=6.3Hz,1.8Hz,1H),3.53(s,3H),2.88(t,J=7.5Hz,2H ),2.43(s,3H),2.27–1.70(m,11H),1.47–1.26(m,3H),1.19(s,3H),1.06(s, 3H),1.04(m,3H),1.02(s,3H),0.98(s,3H),0.96–0.91(m,1H),0.89(s,3H).

[0237] Example 54: Synthesis of Compound 4M

[0238] The synthesis method was the same as that of Example 45, except that p-tert-butylbenzaldehyde was replaced with p-chlorobenzaldehyde to obtain a light yellow solid compound 4M (713 mg, 1.22 mmol) in a yield of 58.3%;

[0239] 1 H NMR(400MHz,Chloroform-d)δ8.18(d,J=8.6Hz,2H),7.55–7.45(m,3H),6.62(d ,J=10.0Hz,1H),6.42(d,J=10.0Hz,1H),5.67(dd,J=6.7,1.7Hz,1H),3.53(s,3 H),2.51(s,3H),2.26–1.97(m,5H),1.88–1.73(m,4H),1.44–1.27(m,3H),1.20 (s,3H),1.07(s,3H),1.04(s,3H),1.00(s,3H),0.97–0.92(m,1H),0.90(s,3H).

[0240] Example 55: Synthesis of Compound 4Q

[0241] The synthesis method was the same as that of Example 45, except that p-tert-butylbenzaldehyde was replaced with cyclopropylcarboxaldehyde to obtain a light yellow solid compound 4Q (552 mg, 1.08 mmol) in a yield of 51.3%;

[0242] 1H NMR (400 MHz, Chloroform-d) δ 7.34 (s, 1H), 6.55 (d, J = 10.0 Hz, 1H), 6.36 (d, J = 10.0 Hz, 1H), 5.61 (dd, J = 6.8, 1.8 Hz, 1H), 3.53 (s, 3H), 2.40 (s, 3H), 2.30 - 1.96 (m, 6H), 1.85 - 1.68 (m, 4H), 1.47 - 1.36 (m, 2H), 1.32 - 1.21 (m, 3H), 1.19 (s, 3H), 1.17 - 1.10 (m, 2H), 1.06 (s, 3H), 1.01 (d, J = 1.8 Hz, 3H), 0.98 (s, 3H), 0.96 - 0.90 (m, 1H), 0.89 (s, 3H).

[0243] Example 56: Synthesis of compound 4U

[0244] The synthesis was carried out as in Example 45, except that p-tert- butylbenzaldehyde was replaced by isobutyraldehyde, to give compound 4U (449 mg, 0.87 mmol) as a light yellow solid in 41.5% yield;

[0245] 1 H NMR (400 MHz, Chloroform-d) δ 7.34 (s, 1H), 6.55 (d, J = 10.0 Hz, 1H), 6.36 (d, J = 10.0 Hz, 1H), 5.61 (dd, J = 6.8, 1.8 Hz, 1H), 3.53 (s, 3H), 2.40 (s, 3H), 2.30 - 1.96 (m, 6H), 1.85 - 1.68 (m, 4H), 1.47 - 1.36 (m, 2H), 1.32 - 1.21 (m, 3H), 1.19 (s, 3H), 1.17 - 1.10 (m, 2H), 1.06 (s, 3H), 1.01 (d, J = 1.8 Hz, 3H), 0.98 (s, 3H), 0.96 - 0.90 (m, 1H), 0.89 (s, 3H).

[0246] Example 57: Synthesis of compound 4W

[0247] The synthesis was carried out as in Example 45, except that p-tert- butylbenzaldehyde was replaced by isobutyraldehyde, to give compound 4U (449 mg, 0.87 mmol) as a light yellow solid in 41.5% yield;

[0248] 1H NMR(400MHz,Chloroform-d)δ9.52(s,2H),9.34(s,1H),7.55(s,1H),6.67(d, J=10.0Hz,1H),6.43(d,J=9.9Hz,1H),5.69(dd,6.4Hz,1.8Hz,1H),3.54(s,3H) ,2.53(s,3H),2.32–1.97(m,5H),1.89–1.70(m,4H),1.45–1.27(m,3H),1.20( s,3H),1.08(s,3H),1.04(s,3H),1.01(s,3H),0.98–0.92(m,1H),0.90(s,3H).

[0249] Example 58: Synthesis of Compound 4F-1

[0250] Synthesis of compound 4F-1: (1) Sodium hydride (4.0 g, 100 mmol) and hexamethylphosphoric acid triamide 50 mL were added to a 100 mL eggplant-shaped flask, stirred for 20 min under N2 protection, and propanethiol (3.8 g, 50 mmol) was slowly added dropwise at 0°C. After the addition was complete, the mixture was reacted at room temperature for 2 h and allowed to stand for use; (2) Compound 4F (2.0 g, 3.64 mmol) was added to a 100 mL eggplant-shaped flask, and 10 mL of the reaction solution of step (1) was added under N2 protection. The mixture was reacted at room temperature for 12 h. After the reaction was completed, water was added at 0°C to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phases were combined, and the organic layer was washed with saturated copper sulfate solution 4 times, washed with water, washed with saturated brine, and dried over anhydrous sodium sulfate. The solvent was concentrated, and the residue was subjected to silica gel column chromatography to obtain white solid compound 4F-1 (1.07 g, 2.00 mmol) with a yield of 55.0%;

[0251] 1 H NMR(400MHz, CDCl3)δ11.28(s,1H),8.39–8.06(m,2H),7.69–7.38(m,4H),6. 56(d,J=10.0Hz,1H),6.29(d,J=9.9Hz,1H),5.65(dd,J=6.8,1.7Hz,1H),2.44 –2.22(m,6H),2.14(td,J=13.9,4.1Hz,1H),2.03–1.73(m,5H),1.53–1.38(m ,2H),1.28(m,4H),1.09(s,3H),1.06(s,3H),1.03(s,3H),1.02–0.91(m,4H).

[0252] Example 59: Synthesis of Compound 2-22

[0253] Synthesis of compound 1A-2: 1A-1 (4.7 g, 100 mmol), 50 mL of toluene, and diphenylphosphoryl azide (DPPA, 2.75 g, 100 mmol) were added to a 100 mL eggplant-shaped flask. The mixture was reacted at 110°C for 8 h under N2 protection. After the reaction, the mixture was extracted with ethyl acetate. The organic phases were combined, washed with water and saturated brine, and dried over anhydrous sodium sulfate. The solvent was concentrated, and the residue was purified by silica gel column chromatography to obtain compound 1A-2 (4.2 g, 90 mmol) as a white solid in a yield of 90%; ESI-MS (m / z) = 468.3 [M+H] + .

[0254] Synthesis of compound 2-22: Compound 1A-2 (100 mg, 0.214 mmol) and DIPEA (55 mg, 0.43 mmol) were dissolved in 10 mL of dichloromethane and stirred at room temperature for 30 min. 1-Boc-4-(4-piperidinyl)-piperazine was then added. After the reaction was complete, dichloromethane was removed by rotary evaporation. The crude product was dissolved in ethyl acetate solution in a single-necked flask, and then 4M HCl / ethyl acetate solution (1 mL) was slowly added. The reaction mixture was stirred at room temperature for 3 h, and ethyl acetate was evaporated. Water was then added to the residue, and the solution was alkalized to pH 10, and the target compound precipitated as a solid. The resulting solid was filtered, dried, and purified to obtain the target compound 2-22 (78 mg, 0.124 mmol) with a yield of 58%;

[0255] 1 H NMR (400MHz, CDCl3) δ8.71(d,J=2.0Hz,1H),8.70(d,J=1.9Hz,1H),7.82(d,J=3.0Hz,1H),6.8 4(d,J=10.1Hz,1H),6.68(d,J=9.9Hz,1H),5.84(dd,J=6.9Hz,J=1.9Hz,1H),4.21(s,1H),4.04 –2.88(m,8H),2.77(s,3H),2.72–2.20(m,9H),2.11–1.73(m,7H),1.65–1.49(m,3H),1.44(s,3 H),1.39–1.26(m,3H),1.10(s,3H),1.09(s,3H),1.06(s,3H),1.05(s,3H),1.05–0.93(m,4H).

[0256] Test Example 1: Osteoclastogenesis Inhibitory Activity Test Method

[0257] RAW264.7 cells in the logarithmic growth phase were seeded at a density of 2000 cells / well in a 96-well plate. After incubation at 37°C, 5% CO2 for 12 hours, 50 ng of RANKL and 0.1 mL of different concentrations of tripterine derivatives as shown in Table 1 were added to each well. An equal volume of blank culture medium was added to the control group, with three replicate wells per group. After 96 hours of incubation in the incubator, the culture medium was discarded and TRAP staining solution was added. After 1 hour, the supernatant was discarded, and the osteoclast area was calculated to calculate the IC 50 The results are shown in Table 1.

[0258] Table 1 Inhibitory activity of tripterygium wilfordii derivatives on osteoclastogenesis (μM)

[0259]

[0260]

[0261] As shown in Table 1, all compounds of the present invention have moderate or good inhibitory activity on osteoclast differentiation and are expected to become new drugs for the treatment of osteoporosis.

Claims

1. A tripterygium wilfordii derivative or a pharmaceutically acceptable salt thereof having activity in treating osteoporosis, characterized in that: Any one of the following general formulas I to VIII: R1 is H, C1-C6 straight or branched chain alkyl, cycloalkyl, 5-12 membered heterocyclic group or heterocyclic aromatic group, wherein the heterocyclic group or heterocyclic aromatic group contains 1-3 heteroatoms selected from O, S, and N; R2 and R3 are independently OH, a halogen atom, a C1-C6 straight or branched alkyl group, a cycloalkyl group, a substituted or unsubstituted 5-12 membered heterocyclic group, a heterocyclic aryl group or a substituted phenyl group, and the substituents are independently C1-C6 straight or branched alkyl group or C1-C6 alkoxy group; or R2 and R3 together form a cyclic structure, the formed cyclic structure is a cycloalkyl group, a substituted or unsubstituted phenyl group, a substituted or unsubstituted heterocyclic group or a heterocyclic aromatic group, the substituents are independently OH, a C1-C6 straight-chain or branched alkyl group, a C1-C6 alkoxy group, a halogen atom, a cyano group, or an amino group, the heterocyclic group or heterocyclic aromatic group contains 1-3 heteroatoms selected from O, S, and N; R4 is OH, a halogen atom, a C1-C6 straight or branched alkyl group, a cycloalkyl group, a substituted or unsubstituted 5-12 membered heterocyclic group, a heterocyclic aryl group, or a phenyl group; the substituents are independently C1-C6 straight or branched alkyl group, C1-C6 alkoxy group, phenyl group, cycloalkyl group, a halogen atom, or two adjacent carbon atoms together form a cyclic structure, which may contain 1-3 heteroatoms selected from O, S, and N; R5 and R6 are independently H, C1-C6 straight or branched alkyl, hydroxyl-substituted alkyl, N,N-dimethylamino-substituted alkyl, halogen-substituted alkyl, amino-substituted alkyl, substituted or unsubstituted phenyl, substituted or unsubstituted benzyl, and any one of the following structural formulas; the substituents are independently halogen atoms, C1-C6 straight or branched alkyl, C1-C6 alkoxy, hydroxyl or amino; Or R5, R6 and the commonly connected N atom form a ring structure, and the formed ring structure includes: R7 and R8 are independently H, C1-C6 straight or branched alkyl, hydroxyl-substituted alkyl, N,N-dimethylamino-substituted alkyl, halogen-substituted alkyl, amino-substituted alkyl, substituted or unsubstituted phenyl, substituted or unsubstituted benzyl, and any one of the following structural formulas; the substituents are independently halogen atoms, C1-C6 straight or branched alkyl, C1-C6 alkoxy, hydroxyl or amino; Or R7, R8 and the commonly connected N atom form a ring structure, and the formed ring structure includes:

2. The tripterygium wilfordii derivative or a pharmaceutically acceptable salt thereof having activity in treating osteoporosis according to claim 1, characterized in that: Any one of the following compounds:

3. A method for preparing the tripterine derivative or a pharmaceutically acceptable salt thereof having activity in treating osteoporosis according to claim 2, characterized in that: The following preparation routes are included: Route 1: Preparation of compounds 1A, 1A-1, 1-1 to 1-31, 4-1 to 4-5 Using tripterygium wilfordii 1 as the starting material, it reacts with iodomethane to obtain intermediate 2; Add methanol and hydrochloric acid to intermediate 2, adjust the pH to 1-2, and react at room temperature to obtain intermediate 3; Manganese dioxide is added to intermediate 3 to undergo oxidation reaction to obtain intermediate 4: Intermediate 4 reacts with ethylenediamine under reflux to obtain compound 1A; Compound 1A is used to prepare compound 1A-1, specifically comprising: (1) adding sodium hydride to a reaction flask, adding HMPA under nitrogen protection, slowly adding propyl mercaptan dropwise at 0°C, and stirring at room temperature; (2) adding compound 1A to the reaction flask, adding the reaction solution of step (1) under nitrogen protection, and reacting at room temperature to obtain compound 1A-1; To compound 1A-1, HATU and DIPEA are added and reacted with the corresponding amine at room temperature to obtain the tripterygium derivative represented by the general formula VII, or R1OH is added and reacted at room temperature to obtain the tripterygium derivative represented by the general formula I; In route 1, R1, R5 and R6 are the corresponding groups in the corresponding positions of compounds 1A, 1A-1, 1-1 to 1-31, 4-1 to 4-5; route 2: preparation of compounds 2B and 2B-1 The intermediate 3 was prepared by the method of route 1, and then a base was added and reacted with dibromomethane under nitrogen protection to prepare compound 2B; referring to the method of preparing compound 1A-1 from compound 1A in route 1, compound 2B-1 was prepared from compound 2B; Route 3: Preparation of compounds 2C and 2C-1 The intermediate 3 was prepared by the method of route 1, and then a base was added and reacted with dibromoethane under nitrogen protection to prepare compound 2C; referring to the method of preparing compound 1A-1 from compound 1A in route 1, compound 2C-1 was prepared from compound 2C; Route 4: Preparation of compounds 3A and 3A-1 Intermediate 4 was prepared by the method of route 1, and then reacted with glycine, ammonium acetate, and acetic acid to prepare compound 3A. Referring to the method of preparing compound 1A-1 from compound 1A in route 1, compound 3A was prepared to obtain compound 3A-1. Route 5: Preparation of compounds 1B-1H Intermediate 4 is prepared by the method of route 1, and then reacted with a substituted o-diamino group in anhydrous ethanol to prepare a compound represented by the general formula IX; R2 and R3 are the corresponding groups at the corresponding positions of compounds 1B-1H; Route 6: Preparation of compounds 4C-4W The intermediate 4 is prepared by the method of route 1, and then reacted with a substituted aldehyde compound, ammonium acetate, and acetic acid to obtain a compound of formula VI; R4 is the corresponding group at the corresponding position of compound 4C-4W; Route 7: Preparation of compound 4F-1 Compound 4F is reacted in a system of sodium hydride, hexamethylphosphoric triamide, and propanethiol at room temperature under N2 protection to obtain compound 4F-1, where R4 is the corresponding group at the corresponding position of compound 4F-1; Route 8: Preparation of compound 2-22 Compound 1A-1 was prepared by the method of route 1; compound 1A-1 was reacted with DPPA to prepare compound 1A-2; compound 1A-2 and DIPEA were dissolved in a solvent, 1-Boc-4-(4-piperidinyl)-piperazine was added, and the intermediate 1A-3 was prepared by reaction, and acid was added for deprotection to prepare compound 2-22.

4. A pharmaceutical composition, characterized in that It comprises the tripterygium wilfordii derivative having osteoporosis treating activity according to claim 1 or 2 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

5. The pharmaceutical composition according to claim 4, characterized in that The administration methods of the pharmaceutical composition include injection and oral administration, and the dosage forms include oral administration dosage forms, injection dosage forms, respiratory tract administration dosage forms, and skin administration dosage forms.

6. Use of the tripterine derivative or a pharmaceutically acceptable salt thereof having activity for treating osteoporosis according to claim 1 or 2 in the preparation of a drug for treating osteoporosis.

7. Use of the pharmaceutical composition according to claim 4 or 5 in the preparation of a drug for treating osteoporosis.

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