Stilbene compound, pharmaceutical composition and application of stilbene compound

Abstract

The invention provides a stilbene compound, a pharmaceutical composition and application of the stilbene compound and relates to the technical field of pharmaceutical chemistry, a main framework of the compound is stilbene with ureido, and the stilbene compound is synthesized by expanding different substituent groups on two sides of the ureido and synthesizing a stilbene side framework containing a natural product dominant structure. A series of high-activity stilbene compounds are obtained, and the IC50 value of the high-activity stilbene compounds for inhibiting soluble epoxide hydrolase reaches a nanomole level, which indicates that the stilbene compounds have an efficient inhibiting effect on soluble epoxide hydrolase inhibiting enzyme; therefore, the inhibition effect on inflammation can be realized by inhibiting the soluble epoxide hydrolase inhibitory enzyme, and the application prospect is good.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical chemistry, and in particular to a stilbene compound, a pharmaceutical composition and application thereof. Background Art

[0002] Inflammation is a very common symptom and a complex pathophysiological process involving multiple enzymes, mediators and receptors. The typical physiological characteristics of inflammation are redness, swelling, fever, pain, and even loss of organ function. Existing studies have shown that the arachidonic acid (AA) pathway produces active mediators that regulate a series of different inflammatory processes. Cyclooxygenase (COX), lipoxygenase (LOX) and cytochrome P450 are the three main pathways for regulating arachidonic acid.

[0003] Epoxyeicosatrienoic acids (EETs) produced by the cytochrome P450 pathway are beneficial arachidonic acid metabolites and important signaling molecules in organisms. They have the functions of regulating gene expression, ion transport, anti-apoptosis, vasodilation, and anti-inflammatory. There are many pathways for the degradation of EETs in organisms, among which soluble epoxide hydrolase (sEH) hydrolyzes them into dihydroxydihydroxy

[0004]

[0005] Dihydroxyeicosatrienoic acids (DHETs) are the main metabolic pathway. Decreased EETs concentration and increased DHETs concentration in the body can induce inflammation. Therefore, inhibiting the activity of sEH and thus increasing the concentration of EETs in the body can effectively treat inflammation and related diseases. Current studies have also shown that sEH is closely related to cardiovascular, central nervous system and metabolic diseases.

[0006] So far, AR9281, GSK2256294, and EC5026 have entered the clinical trial stage, but no sEH inhibitor drugs are currently on the market. A large number of compounds in natural products also show inhibitory effects on sEH, such as natural ureas, triterpenes, flavonoids, and phenylpropionic acid. Unfortunately, the IC values ​​of many compounds reported as sEH inhibitors are low. 50 The values ​​were all at the micromolar level and were not valuable for further preclinical studies.

[0007] Therefore, an IC targeting the anti-inflammatory target soluble epoxide hydrolase is urgently needed.50 The values ​​were all at nanomolar levels for novel framework-soluble epoxide hydrolase inhibitory compounds. Summary of the invention

[0008] The present invention provides a stilbene compound, a pharmaceutical composition and its application to solve the IC of existing soluble epoxide hydrolase inhibitory compounds. 50 The problem is that the value is higher and the drugability is poor.

[0009] In a first aspect, the present invention provides a diphenylethylene compound having a general structural formula of the following formula I:

[0010]

[0011] In the formula I, R 1 , R 2 The groups are combined to form methylenedioxy, or independently selected from methoxy, hydroxyl, hydrogen and any one of the following R1-R9 groups, the structural formula of the R1-R9 groups is as follows:

[0012]

[0013] R 3 Any one group selected from methoxy and hydrogen;

[0014] R 4 Selected from C 1-4 Alkyl, tetrahydropyranyl, N-(C 1-4 Any one of 4-(4-(6-(4-(6-(4-(2 ...

[0015] X and Y are each independently selected from any one of methyl, hydrogen, fluorine and chlorine.

[0016] Based on the first aspect, when the urea group of the compound is located at the para position of the diphenylethylene skeleton, the general structural formula is as shown in Formula II:

[0017]

[0018] In the formula II, R 1 , R 2 , R 3 , R 4 , X, and Y are the same as those defined in claim 1.

[0019] Based on the first aspect, when the urea group of the compound is located at the meta position of the diphenylethylene skeleton, and X=Y=H, the general structural formula is as shown in Formula Ш:

[0020]

[0021] In the formula, R 1 , R 2 , R 3 , R 4 , X, and Y are the same as those defined in claim 1.

[0022] Based on the first aspect, the compound according to claim 1 is characterized in that R 1 , R 2 Combined to form methylenedioxy, or independently selected from methoxy, hydrogen, any one of R1-R9;

[0023] R 3 Any one group selected from methoxy and hydrogen;

[0024] R 4 Any one group selected from isopropyl, isobutyl, tetrahydropyranyl, N-acetylpiperidinyl, N-propionylpiperidinyl and 4-trifluoromethoxyphenyl.

[0025] Based on the first aspect, X of the compound is selected as hydrogen, and Y of the compound is selected as hydrogen.

[0026] Based on the first aspect, the compound R 1 , R 2 Select methylenedioxy, R 3 Select hydrogen, R 4 is tetrahydropyranyl, X is hydrogen, and Y is hydrogen; or

[0027] The R 1 Select methoxy, R 2 Select hydrogen, R 3 is methoxy, R 4 is tetrahydropyranyl, X is hydrogen, and Y is hydrogen; or

[0028] The R 1 Select any one of the groups R1-R9, R 2 Select hydrogen, R 3 Select hydrogen, R 4 is tetrahydropyranyl, X is hydrogen, and Y is hydrogen; or

[0029] The R 2 Select any one of the groups R1-R9, R 1 Select hydrogen, R 3 Select hydrogen, R 4 Select tetrahydropyranyl, X is hydrogen, and Y is hydrogen.

[0030] Furthermore, the structural formula of the compound is:

[0031]

[0032] Based on the first aspect, the specific structural formula of the compound is:

[0033]

[0034]

[0035] In a second aspect, the present invention provides a distilbene compound pharmaceutical composition, which comprises at least one of the compounds described in any one of the first aspect above, their isotope-labeled substances, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds thereof.

[0036] In a third aspect, the present invention provides a use of the diphenylethylene compound as described in any one of the first aspect or the diphenylethylene compound pharmaceutical composition as described in the second aspect in the preparation of anti-inflammatory drugs.

[0037] The present invention includes the following advantages:

[0038] (1) The present invention provides a stilbene compound, which is a derivative of a novel stilbene skeleton, and the IC50 of the compound for soluble epoxy compound hydrolases is at the nanomolar level, which is higher than the micromolar level of natural products;

[0039] (2) The skeleton of the diphenylethylene compound prepared by the present invention contains the dominant structure of natural products - diphenylethylene, which can improve the drugability of the soluble epoxy compound hydrolase inhibitory drug composition. DETAILED DESCRIPTION

[0040] The following examples are provided for a better understanding of the present invention, but are not intended to limit the best mode of implementation, nor to limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior arts shall fall within the protection scope of the present invention.

[0041] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the prior art can be used. The various chemicals used in the present invention are commercially available, and the nuclear magnetic resonance and mass spectrometers used to determine the structures of the following compounds are provided by the Analysis and Testing Center of Beijing Institute of Technology.

[0042] The first aspect of the present invention provides a diphenylethylene compound, wherein the diphenylethylene compound has a general structure as shown in Formula I below:

[0043]

[0044] In the formula I, R 1 , R 2 The groups are combined to form methylenedioxy, or independently selected from methoxy, hydroxyl, hydrogen and any one of the following R1-R9 groups, the structural formula of the R1-R9 groups is as follows:

[0045]

[0046] R 3 Any one group selected from methoxy and hydrogen;

[0047] R 4 Selected from C 1-4 Alkyl, tetrahydropyranyl, N-(C 1-4 Any one of 4-(4-(6-(4-(6-(4-(2 ...

[0048] X and Y are each independently selected from any one of methyl, hydrogen, fluorine and chlorine.

[0049] C 1-4 Alkyl refers to a straight or branched saturated monovalent hydrocarbon group having 1 to 4 carbon atoms. 1-4 The alkyl group means a straight chain or branched chain alkyl group having 1, 2, 3 or 4 carbon atoms, and the alkyl group is, for example, methyl, ethyl, propyl, butyl, isopropyl, isobutyl, sec-butyl, tert-butyl or the like.

[0050] C 1-4 Alkylformyl represents RC(=O)-, where R is C 1-4 The alkylformyl group is, for example, acetyl, propionyl, butyryl, valeryl, isobutyryl, and the like.

[0051] In the second aspect, an embodiment of the present invention further provides a distilbene compound pharmaceutical composition, which comprises at least one of the compounds of any one of the above-mentioned first aspect, its isotope-labeled substances, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds.

[0052] In a third aspect, an embodiment of the present invention further provides a use of the diphenylethylene compound described in the first aspect or the diphenylethylene compound pharmaceutical composition described in the second aspect in the preparation of an anti-inflammatory drug.

[0053] An embodiment of the present invention also provides a method for preventing and / or treating inflammation-related diseases, comprising administering to a patient a preventive or therapeutically effective amount of at least one of the compounds of formula (I), its racemates, stereoisomers, tautomers, isotope-labeled substances, solvates, polymorphs, pharmaceutically acceptable salts, prodrug compounds thereof, or pharmaceutical compositions.

[0054] In some alternative embodiments, the patient is a mammal, preferably a human.

[0055] In the embodiments of the present invention, the pharmaceutical composition also includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents and absorption delaying agents for pharmaceutically active substances, which are reagents and materials known in the art. In addition to any conventional media or agents that are incompatible with the compound, any conventional media or agents can be considered for use in the pharmaceutical composition, and supplementary compounds can also be added to the composition.

[0056] In the embodiments of the present invention, various pharmaceutically acceptable acids can form pharmaceutically acceptable salts on the basic nitrogen of the compound. The acid can be an inorganic acid or an organic acid; the inorganic acid is hydrochloric acid, sulfuric acid, hydrobromic acid, hydroiodic acid or phosphoric acid; the organic acid is acetic acid, malonic acid, methanesulfonic acid, succinic acid, p-toluenesulfonic acid, citric acid, maleic acid, fumaric acid, malic acid or citric acid.

[0057] The prodrug compounds in this specification are administered with a certain structure that is not disclosed, but are metabolized or converted into the compounds disclosed in this article in the human body and exert pharmacological effects as active ingredients. The conventional method for preparing prodrugs can be found in "Design of Prodrugs" (H.Bundgaad, Elsevier, 1985).

[0058] The patient refers to any animal including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates in the present embodiment, and most preferably humans.

[0059] The therapeutically effective amount refers to the amount of active compound or drug that causes the biological or medical response that the researcher, veterinarian, physician or other clinician is seeking in a tissue, system, animal, individual or human, and includes one or more of the following: (1) Preventing disease: for example, preventing a disease, disorder or condition in an individual who is susceptible to the disease, disorder or condition but has not yet experienced or developed the pathology or symptoms of the disease. (2) Inhibiting disease: for example, inhibiting a disease, disorder or condition in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder or condition (i.e., preventing further development of the pathology and / or symptoms). (3) Alleviating disease: for example, alleviating a disease, disorder or condition in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder or condition (i.e., reversing the pathology and / or symptoms).

[0060] In order to enable those skilled in the art to further understand the technical solution of the present invention, a distilbene compound, a pharmaceutical composition and applications thereof provided by the present invention are further described below by way of specific embodiments.

[0061] The following is a brief description of the synthesis method of the stilbene compound of the present invention. In the synthesis examples listed below, the synthesis of the intermediates mainly involves Heck reaction, reduction reaction and acylation reaction.

[0062] Specifically, the synthesis methods of representative specific compounds of the present invention are shown below.

[0063] Synthesis Example 1-Synthesis of Compound C1:

[0064]

[0065] The synthetic route of compound C1-1a is as follows:

[0066]

[0067] Methyltriphenylphosphonium bromide (60mmol, 21.42g) was dissolved in 120mL of dry tetrahydrofuran solution, potassium tert-butoxide (100mmol, 11.2g) was added, and the mixture was reacted at 0°C for 10min under argon protection, and then piperonal (40mmol, 6g) was added to the reaction system, and the mixture was stirred for 5h. After the reaction was completed, saturated ammonium chloride solution was added to the reaction system to terminate the reaction, and the mixture was extracted with dichloromethane, the organic phases were combined, washed with saturated brine, concentrated by rotary evaporation, and dried to obtain crude piperonal. After column chromatography (100-200 mesh silica gel, mobile phase V (petroleum ether): V (ethyl acetate) = 15:1), a colorless transparent liquid compound piperonal (Compound C1-1a) was obtained.

[0068] The synthetic route of compound C1-1b is as follows:

[0069]

[0070] Dissolve piperidine (compound C1-1a) (13.5mmol, 1.71mL) in 50mL dry DMF, add potassium acetate (20.25mmol, 1.99g), palladium acetate (0.621mmol, 0.14g), tetrabutylammonium bromide (18.23mmol, 5.88g) and 4-nitro-1-iodobenzene (11.5mmol, 2.86g). The reaction system was replaced with argon for 5 times and stirred at 80°C for 5h under argon protection. After the reaction was completed, distilled water was added to terminate the reaction, extracted with ethyl acetate, and the upper organic phase was washed with water and saturated brine in turn, concentrated by rotary evaporation, and dried to obtain a solid) (compound C1-1b).

[0071] The synthetic route of compound C1-1 is as follows:

[0072]

[0073] Compound C1-1b was dissolved in ethanol / water (1:1), and ammonium chloride (16.1mmol, 0.86g) and iron powder (25.7mmol, 1.44g) were added, and stirred at 95°C for 5h. After the reaction was completed, it was extracted with ethyl acetate, and the upper organic phase was washed with water, saturated sodium bicarbonate solution, and saturated sodium chloride solution in sequence, concentrated by rotary evaporation, and dried to obtain a solid compound. After column chromatography (200-300 mesh silica gel, mobile phase V (petroleum ether): V (ethyl acetate) = 10:1), a light yellow powder solid compound C1-1 (1.68g, yield 61.2%) was obtained.

[0074] The synthetic route of compound C1 is as follows:

[0075]

[0076] Compound C1-1 (0.418mmol, 0.1g) was dissolved in 10mL of dichloromethane, 4-dimethylaminopyridine DMAP (0.84mmol, 0.102g) was added, and finally carbonyldiimidazole CDI (0.84mmol, 0.136g) was added, and the mixture was reacted at room temperature for 6h under an argon atmosphere. After the reaction was completed, the solid was concentrated by rotary evaporation. The obtained solid compound was redissolved in dry DMF, isobutylamine (1.045mmol, 0.1mL) was added, and stirred at room temperature for 5h. After the reaction was completed, distilled water was added to the reaction solution to terminate the reaction, and the mixture was extracted twice with ethyl acetate. The organic phases were combined and washed with water, saturated ammonium chloride solution, and saturated sodium chloride solution in turn, concentrated by rotary evaporation, and dried to obtain a light yellow solid (compound C1) (0.103g, yield 72.84%).

[0077] HRMS(ESI)(M+H) + m / z 339.1696,calcd for C 20 H 23 N2O3 + 339.1703.mp237.1-239.6℃

[0078] 1 H NMR (400MHz, DMSO-d6) δ8.48(s,1H),7.39(q,4H),7.23(d,J=1.7Hz,1H),7.00(t,2H),6.98(dd,J=8.1,1.7Hz,1H),6.89(d,J =8.0Hz,1H),6.20(t,J=5.9Hz,1H),6.02(s,2H),2.92(t,J=6.3Hz,2H),1.69(dt,J=13.4,6.7Hz,1H),0.87(d,J=6.7Hz,6H). 13CNMR(100MHz,DMSO-d6)δ155.59,148.31,147.02,140.46,132.51,130.50,12 7.21,126.12,121.57,118.02,108.85,105.55,101.45,46.98,28.94,20.48.

[0079] Example 2-Synthesis of Compound C2:

[0080]

[0081] The synthetic route of compound C2 is as follows:

[0082]

[0083] Compound C1-1 (0.418mmol, 0.1g) was dissolved in 10mL of dichloromethane, 4-dimethylaminopyridine (0.84mmol, 0.102g) was added, and finally carbonyldiimidazole (0.84mmol, 0.136g) was added, and the mixture was reacted at room temperature for 6h under an argon atmosphere. After the reaction was completed, the mixture was concentrated by rotary evaporation to obtain a solid. The obtained solid compound was redissolved in dry DMF, 4-aminomethyltetrahydropyran (1.045mmol, 0.12mL) was added, and the mixture was stirred at room temperature for 5h. After the reaction was completed, distilled water was added to the reaction solution to terminate the reaction, and the mixture was extracted twice with ethyl acetate. The organic phases were combined and washed with water, saturated ammonium chloride solution, and saturated sodium chloride solution in turn, concentrated by rotary evaporation, and dried to obtain a light yellow solid (compound C2) (0.122g, yield 76.73%). HRMS (ESI) (M+H) + m / z 381.1801,calcd for C 22 H 25 N2O4 + 381.1809.mp257.7-259.3℃

[0084] 1H NMR (400MHz, DMSO-d6) δ8.51 (s, 1H), 7.44–7.35 (m, 4H), 7.22 (d, J = 1.7Hz, 1H), 7. 07–6.94(m,3H),6.89(d,J=8.0Hz,1H),6.25(t,J=5.9Hz,1H),6.02(s,2H),3.85( ddd,J=11.3,4.5,1.9Hz,2H),3.30–3.16(m,2H),3.00(t,J=6.2Hz,2H),1.64(ddt ,J=11.3,8.2,4.2Hz,1H),1.56(ddd,J=13.0,4.0,1.9Hz,2H),1.25–1.10(m,2H). 13 C NMR(100MHz,DMSO-d6)δ155.64,148.30,147.03,140.41,132.50,130.55,127.20,12 7.06,126.15,121.57,118.07,108.85,105.56,101.45,67.24,45.24,35.82,30.78.

[0085] Example 3-Synthesis of Compound C3:

[0086]

[0087] The synthetic route of compound C3 is as follows:

[0088]

[0089] Compound C1-1 (0.418mmol, 0.1g) was dissolved in 10mL of dichloromethane, 4-dimethylaminopyridine (0.84mmol, 0.102g) was added, and finally carbonyldiimidazole (0.84mmol, 0.136g) was added, and the mixture was reacted at room temperature for 6h under an argon atmosphere. After the reaction was completed, the solid was concentrated by rotary evaporation. The obtained solid compound was redissolved in dry DMF, isopentylamine (1.045mmol, 0.123mL) was added, and stirred at room temperature for 5h. After the reaction was completed, distilled water was added to the reaction solution to terminate the reaction, and the mixture was extracted twice with ethyl acetate. The organic phases were combined and washed with water, saturated ammonium chloride solution, and saturated sodium chloride solution in turn, concentrated by rotary evaporation, and dried to obtain a white solid (compound C3) (0.117g, yield 79.59%). HRMS (ESI) (M+H) + m / z353.1851,calcd for C 21 H 25 N2O3 +353.1860.mp216.5-218.2℃

[0090] 1 H NMR (400MHz, DMSO-d6) δ8.46(s,1H),7.46–7.31(m,4H),7.22(d,J=1.7Hz,1H),6.98(d,J=14.9Hz,3H),6.89(d,J=8.0Hz,1 H), 6.10 (t, J = 5.7Hz, 1H), 6.02 (s, 2H), 3.16–3.04 (m, 2H), 1.66–1.53 (m, 1H), 1.33 (q, J = 7.1Hz, 2H), 0.89 (d, J = 6.6Hz, 6H). 13 C NMR(100MHz,DMSO-d6)δ155.51,148.31,147.03,140.47,132.52,130.51,127.19,12 7.09,126.13,121.55,118.08,108.84,105.57,101.45,39.25,37.73,25.61,22.87.

[0091] Example 4-Synthesis of Compound C4:

[0092]

[0093] The synthetic route of compound C4-1 is as follows:

[0094]

[0095] Compound C1-1a (1.875mmol, 0.24mL) was dissolved in 5mL of dry DMF, potassium acetate (2.813mmol, 0.28g), palladium acetate (0.0938mmol, 0.021g), tetrabutylammonium bromide (2.531mmol, 0.816g) and 2-fluoro-4-iodoaniline (1.688mmol, 0.4g) were added, and the reaction system was replaced with argon three times and reacted at 80°C for 5h. After the reaction was completed, distilled water was added to the reaction system to terminate the reaction, and after extraction with ethyl acetate, the reaction was washed with water, saturated sodium bicarbonate, and saturated sodium chloride, respectively, concentrated by rotary evaporation, and dried to obtain a dark brown solid compound. After column chromatography (200-300 mesh silica gel, mobile phase V (petroleum ether): V (ethyl acetate) = 10:1), a brown-yellow powder solid (compound C4-1) (0.26g, yield 59.87%) was obtained.

[0096] The synthetic route of compound C4 is as follows:

[0097]

[0098] Compound C4-1 (0.389mmol, 0.1g) was dissolved in 10mL of dichloromethane, 4-dimethylaminopyridine (0.778mmol, 0.095g) was added, and finally carbonyldiimidazole (0.778mmol, 0.126g) was added, and the mixture was reacted at room temperature for 6h under an argon atmosphere. After the reaction, the mixture was concentrated by rotary evaporation to obtain solid compound C4-2. The obtained solid compound C4-2 was redissolved in dry DMF, isobutylamine (0.973mmol, 0.097mL) was added, and stirred at room temperature for 5h. After the reaction, distilled water was added to the reaction solution to terminate the reaction, and the mixture was extracted twice with ethyl acetate. The organic phases were combined and washed with water, saturated ammonium chloride solution, and saturated sodium chloride solution in turn, concentrated by rotary evaporation, and dried to obtain a white solid (compound C4) (0.098g, yield 70.71%). HRMS (ESI) (M+H) + m / z 357.1603,calcd for C 20 H 22 FN2O3 + 357.1609.226.9-229.8℃

[0099] 1 H NMR (400MHz, DMSO-d6) δ8.32(d,J=2.7Hz,1H),8.13(t,J=8.7Hz,1H),7.40(dd,J=13.0,1.9Hz,1H),7.27–7.20(m,2H),7.11–6.96(m ,3H),6.90(d,J=8.0Hz,1H),6.67(t,J=5.8Hz,1H),6.03(s,2H),2.97–2.91(m,2H),1.69(hept,J=6.7Hz,1H),0.88(d,J=6.7Hz,6H). 13 C NMR(100MHz,DMSO-d6)δ155.24,153.29,150.90,148.34,147.32,132.12,131.55,131.48,128.06,127.96,127.77 ,126.02,126.00,123.16,123.14,121.91,120.13,112.39,112.20,108.88,105.63,101.53,46.99,28.87,20.45.

[0100] Example 5-Synthesis of Compound C5:

[0101]

[0102] The synthetic route of compound C5-1 is as follows:

[0103]

[0104] C1-1a (1.875mmol, 0.24mL) was dissolved in 5mL dry DMF, potassium acetate (2.813mmol, 0.28g), palladium acetate (0.0938mmol, 0.021g), tetrabutylammonium bromide (2.531mmol, 0.816g) and 3-fluoro-4-iodoaniline (1.688mmol, 0.4g) were added, and the reaction system was replaced with argon five times and reacted at 80°C for 5h. After the reaction was completed, distilled water was added to the reaction system to terminate the reaction, and after extraction with ethyl acetate, the reaction was washed with water, saturated sodium bicarbonate, and saturated sodium chloride, respectively, concentrated by rotary evaporation, and dried to obtain a dark brown solid compound. After column chromatography (200-300 mesh silica gel, mobile phase V (petroleum ether): V (ethyl acetate) = 10:1), a brown powder solid (compound C5-1) (0.23g, yield 52.96%) was obtained.

[0105] The synthetic route of compound C5 is as follows:

[0106]

[0107] Compound C5-1 (0.389mmol, 0.1g) was dissolved in 10mL of dichloromethane, 4-dimethylaminopyridine (0.778mmol, 0.095g) was added, and finally carbonyldiimidazole (0.778mmol, 0.126g) was added, and the mixture was reacted at room temperature for 6h under an argon atmosphere. After the reaction was completed, the solid was concentrated by rotary evaporation. The obtained solid compound was redissolved in dry DMF, isobutylamine (0.973mmol, 0.097mL) was added, and stirred at room temperature for 5h. After the reaction was completed, distilled water was added to the reaction solution to terminate the reaction, and the mixture was extracted twice with ethyl acetate. The organic phases were combined and washed with water, saturated ammonium chloride solution, and saturated sodium chloride solution in turn, concentrated by rotary evaporation, and dried to obtain a white solid (compound C5) (0.102g, yield 73.60%). HRMS (ESI) (M+H) + m / z357.1593,calcd for C 20 H 22 FN2O3 + 357.1609.mp222.1-225.1℃

[0108] 1H NMR (400MHz, DMSO-d6) δ8.70(s,1H),7.55(t,J=8.8Hz,1H),7.48(dd,J=14.0,2.1Hz,1H),7.24(d,J=1.7Hz,1H),7.08–6.97(m,4H ),6.90(d,J=8.0Hz,1H),6.28(t,J=5.8Hz,1H),6.03(s,2H),2.96–2.90(m,2H),1.70(hept,J=6.7Hz,1H),0.87(d,J=6.8Hz,6H). 13 C NMR (100MHz, DMSO-d6) δ161.47,159.03,155.39,148.37,147.37,141.91,141.79,132.26,128.71,128.66,127.84,127.79 ,121.87,119.20,119.17,117.64,117.51,114.02,113.99,108.87,105.73,104.85,104.57,101.54,46.98,28.88,20.45.

[0109] Example 6-Synthesis of Compound C6:

[0110]

[0111] The synthetic route of compound C6-1a is as follows:

[0112]

[0113] C1-1a (1.875mmol, 0.24mL) was dissolved in 5mL of dry DMF, potassium acetate (2.813mmol, 0.28g), palladium acetate (0.0938mmol, 0.021g), tetrabutylammonium bromide (2.531mmol, 0.816g) and 3-methyl-4-iodonitrobenzene (1.688mmol, 0.444g) were added, and the reaction system was protected by argon and reacted at 80°C for 5h. After the reaction was completed, distilled water was added to the reaction system to terminate the reaction, and after extraction with ethyl acetate, the reaction was washed with water, saturated sodium bicarbonate, and saturated sodium chloride, respectively, concentrated by rotary evaporation, and dried to obtain a dark brown solid compound (C6-1a).

[0114] The synthetic route of compound C6-1 is as follows:

[0115]

[0116] The obtained solid compound C6-1a was dissolved in ethanol / water (1:1), and ammonium chloride (4.22mmol, 0.226g) and iron powder (6.752mmol, 0.377g) were added, and stirred at 95°C for 5h. After the reaction was completed, it was extracted with ethyl acetate, and the upper organic phase was washed with water, saturated sodium bicarbonate solution, and saturated sodium chloride solution in turn, and concentrated by rotary evaporation. After column chromatography (200-300 mesh silica gel, mobile phase V (petroleum ether): V (ethyl acetate) = 10:1), a brown powder solid (compound C6-1) (0.24g, yield 56.13%) was obtained.

[0117] The synthetic route of compound C6 is as follows:

[0118]

[0119] Compound C6-1 (0.395mmol, 0.1g) was dissolved in 10mL of dichloromethane, 4-dimethylaminopyridine (0.79mmol, 0.097g) was added, and finally carbonyldiimidazole (0.79mmol, 0.128g) was added, and the mixture was reacted at room temperature for 6h under an argon atmosphere. After the reaction was completed, the solid was concentrated by rotary evaporation. The obtained solid compound was redissolved in dry DMF, isobutylamine (0.988mmol, 0.098mL) was added, and stirred at room temperature for 5h. After the reaction was completed, distilled water was added to the reaction solution to terminate the reaction, and the mixture was extracted twice with ethyl acetate. The organic phases were combined and washed with water, saturated ammonium chloride solution, and saturated sodium chloride solution in turn, concentrated by rotary evaporation, and dried to obtain a white solid (compound C6) (0.083g, yield 59.62%).

[0120] HRMS(ESI)(M+H) + m / z 353.1851,calcd for C 21 H 25 N2O3 + 353.1860.mp190.6-192.8℃

[0121] 1 H NMR(400MHz, DMSO-d6)δ8.37(s,1H),7.48(d,J=8.5Hz,1H),7.27(d,J=1.7Hz,1H),7.26–7.11(m,3H),7.00(dd,J=8.2,1.7Hz,1H), 6.94–6.83(m,2H),6.16(t,J=5.8Hz,1H),6.02(s,2H),2.92(t,2H),2.33(s,3H),1.69(hept,J=6.7Hz,1H),0.87(d,J=6.7Hz,5H). 13C NMR(100MHz,DMSO-d6)δ155.60,148.30,147.04,140.25,136.29,132.78,129.29,127.45,12 5.80,124.56,121.68,119.45,115.94,108.81,105.82,101.45,46.99,28.95,20.48,20.25.

[0122] Example 7-Synthesis of Compound C7:

[0123]

[0124] The synthetic route of compound C7-1 is as follows:

[0125]

[0126] C1-1a (1.875mmol, 0.24mL) was dissolved in 5mL of dry DMF, potassium acetate (2.813mmol, 0.28g), palladium acetate (0.0938mmol, 0.021g), tetrabutylammonium bromide (2.531mmol, 0.816g) and 2-methyl-4-iodoaniline (1.688mmol, 0.39g) were added, and the reaction system was protected by argon and reacted at 80°C for 5h. After the reaction, distilled water was added to the reaction system to terminate the reaction, and after extraction with ethyl acetate, the reaction was washed with water, saturated sodium bicarbonate, and saturated sodium chloride, respectively, concentrated by rotary evaporation, and dried to obtain a dark brown solid compound. After column chromatography (200-300 mesh silica gel, mobile phase V (petroleum ether): V (ethyl acetate) = 10:1), a brown-yellow powder solid (compound C7-1) (0.23g, yield 53.47%) was obtained.

[0127] The synthetic route of compound C7 is as follows:

[0128]

[0129] Compound C7-1 (0.395mmol, 0.1g) was dissolved in 10mL of dichloromethane, 4-dimethylaminopyridine (0.79mmol, 0.097g) was added, and finally carbonyldiimidazole (0.79mmol, 0.128g) was added, and the mixture was reacted at room temperature for 6h under argon protection. After the reaction was completed, the solid was concentrated by rotary evaporation. The obtained solid compound was redissolved in dry DMF, isobutylamine (0.988mmol, 0.098mL) was added, and stirred at room temperature for 5h. After the reaction was completed, distilled water was added to the reaction solution to terminate the reaction, and the mixture was extracted twice with ethyl acetate. The organic phases were combined and washed with water, saturated ammonium chloride solution, and saturated sodium chloride solution in turn, concentrated by rotary evaporation, and dried to obtain a white solid (compound C7) (0.096g, yield 68.96%).

[0130] HRMS(ESI)(M+H) + m / z 353.1862,calcd for C 21 H 25 N2O3 + 353.1860.mp220.6-222.2℃

[0131] 1 H NMR (400MHz, DMSO-d6) δ7.88(d,J=8.4Hz,1H),7.65(s,1H),7.32(d,J=2.1Hz,1H),7.27(dd,J=8.5,2.2Hz,1H),7.22(d,J=1.7Hz,1H),7.07–6.93(m, 3H),6.89(d,J=8.0Hz,1H),6.64(t,J=5.8Hz,1H),6.02(s,2H),2.93(t,J= 6.2Hz, 2H), 2.20 (s, 3H), 1.70 (hept, J = 6.7Hz, 1H), 0.89 (d, J = 6.7Hz, 6H). 13 C NMR(100MHz,DMSO-d6)δ155.75,148.31,147.04,138.33,132.51,130.96,128.43,127.09,12 6.63,126.33,124.77,121.56,120.32,108.85,105.57,101.45,47.08,28.95,20.54,18.46.

[0132] Example 8-Synthesis of Compound C8:

[0133]

[0134] The synthetic route of compound C8-1a is as follows:

[0135]

[0136] C1-1a (1.875mmol, 0.24mL) was dissolved in 5mL of dry DMF, potassium acetate (2.813mmol, 0.28g), palladium acetate (0.0938mmol, 0.021g), tetrabutylammonium bromide (2.531mmol, 0.816g) and 3-chloro-4-iodinitrobenzene (1.688mmol, 0.478g) were added, and the reaction system was protected by argon and reacted at 80°C for 5h. After the reaction was completed, distilled water was added to the reaction system to terminate the reaction, and after extraction with ethyl acetate, the reaction was washed with water, saturated sodium bicarbonate, and saturated sodium chloride, respectively, concentrated by rotary evaporation, and dried to obtain a dark brown solid (compound C8-1a).

[0137] The synthetic route of compound C8-1 is as follows:

[0138]

[0139] The obtained solid compound C8-1a was dissolved in ethanol / water (1:1), and ammonium chloride (4.22mmol, 0.226g) and iron powder (6.752mmol, 0.377g) were added, and stirred at 95°C for 5h. After the reaction was completed, it was extracted with ethyl acetate, and the upper organic phase was washed with water, saturated sodium bicarbonate solution, and saturated sodium chloride solution in turn, and concentrated by rotary evaporation. After column chromatography (200-300 mesh silica gel, mobile phase V (petroleum ether): V (ethyl acetate) = 10:1), a yellow powder solid (compound C8-1) (0.21g, yield 45.45%) was obtained.

[0140] The synthetic route of compound C8 is as follows:

[0141]

[0142] Compound C8-1 (0.365mmol, 0.1g) was dissolved in 10mL of dichloromethane, 4-dimethylaminopyridine (0.79mmol, 0.097g) was added, and finally carbonyldiimidazole (0.79mmol, 0.128g) was added, and the mixture was reacted at room temperature for 6h under an argon atmosphere. After the reaction was completed, the solid was concentrated by rotary evaporation. The obtained solid compound was redissolved in dry DMF, isobutylamine (0.988mmol, 0.098mL) was added, and stirred at room temperature for 5h. After the reaction was completed, distilled water was added to the reaction solution to terminate the reaction, and the mixture was extracted twice with ethyl acetate. The organic phases were combined and washed with water, saturated ammonium chloride solution, and saturated sodium chloride solution in turn, concentrated by rotary evaporation, and dried to obtain a white solid (compound C8) (0.113g, yield 83.09%).

[0143] HRMS(ESI)(M+H) + m / z 274.0624,calcd for C 15 H 13 ClNO2 + 274.0629.

[0144] 1 H NMR(400MHz,DMSO-d6)δ7.49(d,J=8.6Hz,1H),7.18–7.09(m,2H),6.97(dd,J=8.1,1.7Hz,1H),6 .93–6.85(m,2H),6.62(d,J=2.3Hz,1H),6.55(dd,J=8.5,2.3Hz,1H),6.02(s,2H),5.57(s,2H). 13 C NMR(100MHz,DMSO-d6)δ150.04,148.31,147.04,133.31,132.56,127.53, 126.31,122.94,122.17,121.29,113.88,113.79,108.93,105.65,101.48.

[0145] Example 9-Synthesis of Compound C9:

[0146]

[0147] The synthetic route of compound C9-1a is as follows:

[0148]

[0149] The synthesis steps of compound C9-1a are similar to those of compound C1-1a. For details, please refer to the synthesis steps of compound C1-1a.

[0150] The synthetic route of compound C9-1b is as follows:

[0151]

[0152] The synthesis steps of compound C9-1b are similar to those of compound C1-1b. For details, please refer to the synthesis steps of compound C1-1b.

[0153] The synthetic route of compound C9-1 is as follows:

[0154]

[0155] The synthesis steps of compound C1-1 are similar to those of compound C1-1. For details, please refer to the synthesis steps of compound C1-1.

[0156] The synthetic route of compound C9 is as follows:

[0157]

[0158] Compound C9-1 (0.314mmol, 0.08g) was dissolved in 10mL of dichloromethane, 4-dimethylaminopyridine (0.627mmol, 0.077g) was added, and finally carbonyldiimidazole (0.627mmol, 0.102g) was added, and the mixture was reacted at room temperature for 6h under an argon atmosphere. After the reaction was completed, the solid was concentrated by rotary evaporation. The obtained solid compound was redissolved in dry DMF, isobutylamine (1.568mmol, 0.16mL) was added, and stirred at room temperature for 5h. After the reaction was completed, distilled water was added to the reaction solution to terminate the reaction, and the mixture was extracted twice with ethyl acetate. The organic phases were combined and washed with water, saturated ammonium chloride solution, and saturated sodium chloride solution in turn, concentrated by rotary evaporation, and dried to obtain a light yellow solid (compound C9) (0.084g, yield 75.43%). HRMS (ESI) (M+H) + m / z 355.2007,calcd for C 21 H 27 N2O3 + 355.2016.mp166.6-168.3℃

[0159] 1 H NMR (400MHz, DMSO-d6) δ8.49(s,1H),7.49–7.35(m,4H),7.17(d,J=16.5Hz,1H),7.00(d,J=16.4Hz,1H),6.73(d,J=2.3Hz,2H), 6.38(t,J=2.2Hz,1H),6.20(t,J=5.8Hz,1H),3.77(s,6H),2.97–2.87(m,2H),1.69(hept,J=6.7Hz,1H),0.88(d,J=6.6Hz,6H). 13 C NMR (100MHz, DMSO-d6) δ161.12,155.58,140.85,139.98,130.16,129.30,127.55,126.33,118.02,104.62,99.92,55.65,47.00,28.94,20.47.

[0160] Example 10-Synthesis of Compound C10:

[0161]

[0162] The synthetic route of compound C10 is as follows:

[0163]

[0164] Compound C9-1 (0.2mmol, 0.05g) was added to a 100ml round-bottom flask, and stirred and dissolved with 10ml of anhydrous dichloromethane. CDI (0.4mmol, 0.065g) and DMAP (0.4mmol, 0.05g) were added, and the reaction was continued overnight for 12h. After the reaction, the solvent was dried to obtain a white powder, and DMF (10ml) was added, and stirred and dissolved. 4-aminomethyltetrahydropyran (0.5mmol, 0.06g) was added and the reaction was continued for 6h. After the reaction, distilled water was added to terminate the reaction, and the mixture was extracted with ethyl acetate, the organic phase was separated, washed with saturated brine, the solvent was dried, and recrystallized with ethyl acetate / petroleum ether system to obtain a white solid (compound C10) (0.054g, yield 68.09%).

[0165] HRMS(ESI)(M+H) + m / z 397.2118,calcd for C 23 H 29 N2O4 + 397.2122.mp188.4-190.2℃

[0166] 1 H NMR(400MHz, DMSO-D6)δ:8.57(s,1H),7.57–7.37(m,4H),7.17(d,J=16.4Hz,1H),7.01(d,J= 16.4Hz,1H),6.73(d,J=2.3Hz,2H),6.38(t,J=2.2Hz,1H),6.29(t,J=5.9Hz,1H),3.85(ddd, J=11.3,4.5,1.8Hz,2H),3.77(s,6H),3.27(td,J=11.7,2.1Hz,2H),3.00(t,J=6.2Hz,2H),1 .65(dqt,J=10.9,6.9,3.3Hz,1H),1.56(ddd,J=12.9,4.0,1.9Hz,2H),1.25–1.10(m,2H).13C NMR(100MHz,DMSO)δ161.11,155.62,140.81,139.97,130.17,129.62,129.29,12 7.54,126.33,118.03,106.89,104.60,99.91,67.24,55.64,45.23,35.82,30.78.

[0167] Example 11-Synthesis of Compound C11:

[0168]

[0169] The synthetic formula of compound C11-1 is as follows:

[0170]

[0171] In an ice bath, 4-aminomethylpiperidine (2.4mmol, 0.274g), 20ml of anhydrous dichloromethane, pyridine (2.64mmol, 0.213ml), DMAP (0.24mmol, 0.29g) were added to a 100ml round-bottom flask in sequence, and then propionic anhydride (2.64mmol, 0.339ml) was dissolved in 10ml of anhydrous dichloromethane and added dropwise to the reaction system. The reaction was terminated after 1h under TLC iodine display monitoring, and the solvent was spin-dried to obtain a colorless transparent liquid mixture. After column chromatography (200-300 mesh silica gel), the mobile phase was (V (DCM): V (ethanol): V (ammonia water) = 10: 1: 0.1), and a colorless transparent liquid 1-(4-(aminomethyl)piperidin-1-yl)propane-1-one (compound C11-1) (0.18g, yield 44.05%) was obtained for use.

[0172] The synthetic route of compound C11 is as follows:

[0173]

[0174] Compound C9-1 (0.2mmol, 0.05g) was added to a 100ml round-bottom flask, and stirred and dissolved with 10ml of anhydrous dichloromethane, and CDI (0.4mmol, 0.065g) and DMAP (0.4mmol, 0.05g) were added, and the reaction was continued overnight for 12h. After the reaction, the solvent was dried to obtain white powder C11-2, and DMF (10ml) was added, and stirred to dissolve, and the spare C11-1 (0.5mmol, 0.085g) was added, and the reaction was continued for 6h. After the reaction, distilled water was added to terminate the reaction, and the mixture was extracted with ethyl acetate, and the organic phase was separated, washed with saturated brine, the solvent was dried, and recrystallized with ethyl acetate / petroleum ether system to obtain a white solid (compound C11) (0.07g, yield 77.7%).

[0175] HRMS(ESI)(M+H) + m / z 452.2505,calcd for C 26 H 34 N3O4 + 452.2553.mp80.1-81.5℃

[0176] 1H NMR (400MHz, DMSO-D6) δ: 8.53 (s, 1H), 7.57–7.37 (m, 4H), 7.17 (d, J = 16.4Hz, 1H), 7.01 (d, J=16.3Hz,1H),6.73(d,J=2.3Hz,2H),6.38(t,J=2.2Hz,1H),6.28(t,J=5.9Hz,1H),4.39(d ,J=13.0Hz,1H),3.85(d,J=13.7Hz,1H),3.77(s,5H),3.04–2.89(m,3H),2.50(m,1H),2.2 9(q,J=7.4Hz,2H),1.71–1.61(m,3H),1.08(td,J=12.7,4.1Hz,1H),0.98(t,J=7.4Hz,4H). 13 C NMR(100MHz,DMSO)δ:171.44,161.11,155.60,140.78,139.96,130.20,129.29,127.54,126 .35,118.04,104.61,99.91,55.64,45.14,44.80,41.39,36.88,30.52,29.73,26.09,10.01.

[0177] Example 12-Synthesis of Compound C12:

[0178]

[0179]

[0180] The synthetic route of compound C12 is as follows:

[0181]

[0182] Dissolve methyltriphenylphosphonium bromide (60mmol, 21.42g) in dry tetrahydrofuran solution, add potassium tert-butoxide (100mmol, 11.2g), stir at 0℃ for 10min under argon protection, add 3-hydroxy-4-methoxybenzaldehyde (40mmol, 6.086g) to the reaction system, and continue stirring for 5h. After the reaction is completed, add saturated ammonium chloride solution to the reaction system, extract with dichloromethane three times, combine the organic phases, wash with saturated sodium chloride solution, concentrate by rotary evaporation, and obtain colorless transparent liquid compound 3-hydroxy-4-methoxystyrene by column chromatography (200-300 mesh silica gel, mobile phase V (petroleum ether): V (ethyl acetate) = 15:1) for use. Using the method of compound C1, 4-iodoaniline (2.283mmol, 0.5g) was reacted with CDI and DMAP to connect the carbonyl imidazole group and then reacted with isobutylamine. After column chromatography (200-300 mesh silica gel, mobile phase V (petroleum ether): V (ethyl acetate) = 10:1), it was dried to obtain a white solid compound C12-1 (0.584g, 80.41%). 1HNMR (400MHz, DMSO-d6) δ8.50 (s, 1H), 7.56–7.48 (m, 2H), 7.28–7.20 (m, 2H), 6.20 (t, J = 5.8Hz, 1H), 2.91 (dd, J = 6.7, 5.8Hz, 2H), 1.77–1.59 (m, J = 6.7Hz, 1H), 0.86 (d, J = 6.7Hz, 6 H). Using the method for synthesizing compound C1, compound C12-1 (0.5mmol, 0.16g) and 3-hydroxy-4-methoxystyrene (0.1mL, 0.63mmol) underwent Heck coupling reaction, and the resulting mixture was purified by column chromatography (200-300 mesh silica gel, V (dichloromethane): V (ethanol) = 10:0.1) to obtain a white solid compound (compound C12) (0.086g, 50.59%).

[0183] HRMS(ESI)(M+H) + m / z 341.1865,calcd for C 20 H 25 N2O3 + 341.1860.mp209.7-212.3℃.

[0184] 1H NMR (400MHz, DMSO-d6) δ8.96(s,1H),8.49(s,1H),7.39(q,J=8.9Hz,4H),7.00(d,J=1.9Hz,1H),6.97–6.83(m,4 H), 6.21 (t, J = 5.9Hz, 1H), 3.77 (s, 3H), 2.92 (t, J = 6.3Hz, 2H), 1.69 (hept, J = 6.7Hz, 1H), 0.87 (d, J = 6.7Hz, 7H). 13 C NMR(100MHz,DMSO-d6)δ155.62,147.81,147.07,140.31,131.01,130.65,12 7.12,126.37,118.47,118.02,113.16,112.71,56.11,46.99,28.94,20.48.

[0185] Example 13-Synthesis of Compound C13:

[0186]

[0187] The synthetic route of compound C13-1 is as follows:

[0188]

[0189] After adding p-nitrostyrene (6.705mmol, 1g), iron powder (26.82mmol, 1.5g), and ammonium chloride (16.733mmol, 0.9g) to a 100ml round-bottom flask, 30ml of water was added and stirred, and refluxed at 95°C for 2h. After the reaction was completed, it was extracted with ethyl acetate, and the residue was removed by suction filtration. The liquid was washed with saturated sodium bicarbonate, and the organic phase was washed with saturated brine after separation, and the solvent was dried by spin drying. Then, column chromatography (200-300 mesh silica gel, mobile phase V (petroleum ether): V (ethyl acetate) = 10:1) was performed to obtain colorless liquid p-aminostyrene (0.7g, yield 87.72%). Add p-aminostyrene (5.875mmol, 0.7g) and DMAP (11.7mmol, 1.435g) to a 100ml round-bottom flask, add 20ml dichloromethane DCM and stir to dissolve, finally add CDI (11.71mmol, 1.9g), react overnight, spin dry the solvent, add 20ml N,N-dimethylformamide DMF, add 4-aminomethyltetrahydropyran (20.1mmol, 2.315g), react for 6h. Add water, extract with ethyl acetate, wash the organic phase with saturated brine, spin dry, and recrystallize from ethyl acetate / petroleum ether to obtain a white solid C13-1a (1.02g, yield 65.4%). The obtained white solid C13-1a (1.536mmol, 0.4g), m-iodoaniline (1.706mmol, 0.374g), tetrabutylammonium bromide (2.303mmol, 0.742g), potassium acetate (2.559mmol, 0.251g), palladium acetate (0.0853mmol, 0.02g) were added to a pressure bottle in sequence, and 10ml of DMF was added. Under argon protection, the mixture was reacted at 80°C for 5h. After the reaction, distilled water was added to terminate the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine and dried. The solid was subjected to column chromatography (200-300 mesh silica gel, V (DCM): V (methanol) = 10:0.1) to obtain a pink solid (compound C13-1) (0.2g, yield 37.04%).

[0190] The synthetic route of compound C13 is as follows:

[0191]

[0192] Add 2-picolinic acid (0.2736mmol, 0.034g) to a 100ml round-bottom flask, use 30ml DMF, add diisopropylethylamine DIEA (0.684mmol, 0.088g), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate HATU (0.2736mmol, 0.104g), stir to dissolve, and finally add compound C13-1 (0.228mmol, 0.08g). After the reaction is completed, add distilled water to terminate the reaction, extract with ethyl acetate, separate the organic phase, wash with saturated ammonium chloride, saturated sodium bicarbonate, and saturated brine respectively, spin dry the solvent, and recrystallize with ethyl acetate / petroleum ether system to obtain a white solid (compound C13) (0.084g, yield 80.77%).

[0193] HRMS(ESI)(M+H) + m / z 457.2228,calcd for C 27 H 29 N4O3 + 457.2234.mp232.4-235.2℃

[0194] 1 H NMR (400MHz, DMSO-d6) δ10.60(s,1H),8.76(d,J=4.7Hz,1H),8.51(s,1H),8.18(d,J=7. 8Hz,1H),8.14–8.05(m,2H),7.79(d,J=7.7Hz,1H),7.69(dd,J=7.6,4.8Hz,1H),7.48(d ,J=8.4Hz,2H),7.44–7.28(m,4H),7.11(q,J=16.4Hz,2H),6.24(t,J=5.9Hz,1H),3.89– 3.81(m,2H),3.27–3.13(m,2H),3.00(t,J=6.2Hz,2H),1.57(m,3H),1.25–1.07(m,2H).

[0195] 13 C NMR (100MHz, DMSO) δ162.93,155.66,150.32,148.92,140.89,139.13,138.65,138.46,130.14,129.46, 129.14,127.57,127.44,126.15,122.82,122.62,119.60,118.11,118.04,67.25,45.24,35.84,30.80.

[0196] Example 14-Synthesis of Compound C14:

[0197]

[0198] The synthetic route of compound C14 is as follows:

[0199]

[0200] Add 2-pyridineacetic acid hydrochloride (0.2736mmol, 0.047g) to a 100ml round-bottom flask, use 30ml DMF, add DIEA (0.684mmol, 0.088g), HATU (0.2736mmol, 0.104g), stir to dissolve, and finally add compound C13-1 (0.228mmol, 0.08g). After the reaction is completed, add distilled water to terminate the reaction, extract with ethyl acetate, separate the liquid to obtain the organic phase, wash with saturated ammonium chloride, saturated sodium bicarbonate, and saturated brine respectively, spin dry the solvent, and recrystallize with ethyl acetate / petroleum ether system to obtain a white solid (compound C14) (0.062g, yield 57.94%).

[0201] HRMS(ESI)(M+H) + m / z 471.2396,calcd for C 28 H 31 N4O3 + 471.2391.mp193.7-195.2℃

[0202] 1 H NMR (400 MHz, DMSO-d6) 1 H NMR (400MHz, DMSO-d6) δ10.25(s,1H),8.51(d,J=4.6Hz,2H),7.87–7.72(m,2H),7.43(dt,J=22.0,8.2Hz,6H),7.33–7.21(m,3H),7.05(s,2H),6. 24(t,J=5.9Hz,1H),3.85(d,J=13.5Hz,4H),3.27(td,J=12.0,2.4Hz,2H) ,3.00(t,J=6.3Hz,2H),1.70–1.52(m,3H),1.17(tt,J=12.2,6.0Hz,2H). 13C NMR (100MHz, DMSO) δ168.71,156.50,155.60,149.45,140.81,139.99,138.39,137.02,130.15,129.47,128.91 ,127.56,126.27,124.46,122.38,121.78,118.60,118.06,117.14,67.24,55.37,46.36,45.25,35.81,30.79.

[0203] Example 15-Synthesis of Compound C15:

[0204]

[0205] The synthetic route of compound C15 is as follows:

[0206]

[0207] Add 3-pyridineacetic acid hydrochloride (0.2736mmol, 0.047g) to a 100ml round-bottom flask, use 30ml DMF, add DIEA (0.684mmol, 0.088g), HATU (0.2736mmol, 0.104g), stir to dissolve, and finally add compound C13-1 (0.228mmol, 0.08g). After the reaction is completed, add distilled water to terminate the reaction, extract with ethyl acetate, separate the liquid to obtain the organic phase, wash with saturated ammonium chloride, saturated sodium bicarbonate, and saturated brine respectively, spin dry the solvent, and recrystallize with ethyl acetate / petroleum ether system to obtain a white solid (compound C15) (0.046g, yield 42.99%).

[0208] HRMS(ESI)(M+H) + m / z 471.2371,calcd for C 28 H 31 N4O3 + 471.2391.mp198.2-200.3℃

[0209] 1H NMR (400MHz, DMSO-d6) δ10.29(s,1H),8.65(s,1H),8.55(d,J=2.3Hz,1H),8.47(dd,J= 4.8,1.7Hz,1H),7.82–7.74(m,2H),7.50–7.34(m,6H),7.31–7.21(m,2H),7.04(s,2H), 6.34(t,J=5.9Hz,1H),3.85(ddd,J=11.4,4.5,1.9Hz,2H),3.73(s,2H),3.27(td,J=11 .7,2.1Hz,2H),3.00(t,J=6.2Hz,2H),1.71–1.53(m,3H),1.18(qd,J=12.1,4.7Hz,2H).

[0210] 13 C NMR (100MHz, DMSO) δ169.07,155.66,150.68,148.27,140.88,139.90,138.42,137.23,132.12,130.08, 129.49,128.96,127.56,126.18,123.85,121.86,118.64,118.00,117.19,67.24,45.24,35.83,30.80.

[0211] Example 16-Synthesis of Compound C16:

[0212]

[0213] The synthetic route of compound C16 is as follows:

[0214]

[0215] Add 4-pyridineacetic acid hydrochloride (0.2736mmol, 0.047g) to a 100ml round-bottom flask, use 30ml DMF, add DIEA (0.684mmol, 0.088g), HATU (0.2736mmol, 0.104g), stir to dissolve, and finally add compound C13-1 (0.228mmol, 0.08g). After the reaction is completed, add distilled water to terminate the reaction, extract with ethyl acetate, separate the liquid to obtain the organic phase, wash with saturated ammonium chloride, saturated sodium bicarbonate, and saturated brine respectively, spin dry the solvent, and recrystallize with ethyl acetate / petroleum ether system to obtain a white solid (compound C16) (0.038g, yield 35.51%).

[0216] HRMS(ESI)(M+H) +m / z 471.2386,calcd for C 28 H 31 N4O3 + 471.2391.mp212.6-214.8℃

[0217] 1 H NMR (400MHz, DMSO-d6) δ10.27(s,1H),8.57–8.45(m,3H),7.79(t,J=1.8Hz,1H),7.50–7.23(m,9H),7.05(s,2H),6.27(t,J=6.0Hz,1H),3.85(d dd,J=11.2,4.5,1.9Hz,2H),3.72(s,2H),3.27(td,J=11.6,2.0Hz,2H),3.00(t,J=6.2Hz,2H),1.72–1.52(m,3H),1.18(qd,J=12.1,4.6Hz,2H). 13 C NMR (100MHz, DMSO) δ168.35,155.61,149.96,145.17,140.85,139.81,138.44,130.11,129.51,128 .98,127.57,126.19,125.13,121.92,118.65,118.04,117.22,67.24,45.24,42.90,35.82,30.79.

[0218] Example 17-Synthesis of Compound C17:

[0219]

[0220] The synthetic route of compound C17 is as follows:

[0221]

[0222] Add 2-chloropyridine-5-acetic acid (0.2736mmol, 0.047g) to a 100ml round-bottom flask, add 30ml DMF, add DIEA (0.684mmol, 0.088g), HATU (0.2736mmol, 0.104g), stir to dissolve, and finally add compound C13-1 (0.228mmol, 0.08g). After the reaction is completed, add distilled water to terminate the reaction, extract with ethyl acetate, separate the liquid to obtain the organic phase, wash with saturated ammonium chloride, saturated sodium bicarbonate, and saturated brine respectively, spin dry the solvent, and recrystallize with ethyl acetate / petroleum ether system to obtain a white solid (compound C17) (0.046g, yield 41.74%).

[0223] HRMS(ESI)(M+H) + m / z 505.1972,calcd for C 28 H 30 ClN4O3 + 505.2001.mp207.7-208.9℃

[0224] 1 H NMR (400MHz, DMSO-d6) δ10.27(s,1H),8.54(s,1H),8.37(d,J=2.5Hz,1H),7.87–7.77(m,2H ),7.45(ddd,J=28.3,13.0,8.4Hz,6H),7.33–7.22(m,2H),7.05(s,2H),6.27(t,J=5.8Hz,1H ),3.85(ddd,J=11.4,4.7,1.9Hz,2H),3.75(s,2H),3.27(td,J=11.7,2.1Hz,2H),3.00(t,J =6.1Hz,2H),1.71–1.61(m,1H),1.56(dd,J=14.2,3.4Hz,2H),1.18(qd,J=12.0,4.6Hz,2H). 13 C NMR (100MHz, DMSO) δ168.69,155.60,150.84,149.05,141.15,140.84,139.81,138.42,131.67,130.09, 129.53,128.95,127.58,126.18,124.32,121.91,118.63,118.02,117.18,67.24,45.24,35.81,30.78.

[0227] Example 18-Synthesis of Compound C18:

[0228]

[0229] The synthetic route of compound C18 is as follows:

[0230]

[0231] Add N-indoleacetic acid (0.2736mmol, 0.048g) to a 100ml round-bottom flask, use 30ml DMF, add DIEA (0.684mmol, 0.088g), HATU (0.2736mmol, 0.104g), stir to dissolve, and finally add compound C13-1 (0.228mmol, 0.08g). After the reaction is completed, add distilled water to terminate the reaction, extract with ethyl acetate, separate the liquid to obtain the organic phase, wash with saturated ammonium chloride, saturated sodium bicarbonate, and saturated brine respectively, spin dry the solvent, and recrystallize with ethyl acetate / petroleum ether system to obtain a pink solid (compound C18) (0.038g, yield 35.51%).

[0232] HRMS(ESI)(M+Na) + m / z 531.2370,calcd for C 31 H 32 N4O3Na + 531.2367.mp228.5-230.2℃

[0233] 1 H NMR (400MHz, DMSO-d6) δ10.44(s,1H),8.59(s,1H),7.83(t,J=1.8Hz,1H),7.57(d,J=7.8Hz,1 H),7.49–7.36(m,7H),7.35–7.23(m,2H),7.13(ddd,J=8.2,7.0,1.2Hz,1H),7.04(d,J=3.3Hz ,3H),6.50–6.42(m,1H),6.30(t,J=5.8Hz,1H),5.08(s,2H),3.85(ddd,J=11.4,4.5,1.8Hz,2 H),3.26(td,J=11.7,2.0Hz,2H),2.99(t,J=6.2Hz,2H),1.71–1.52(m,3H),1.24–1.14(m,2H). 13 C NMR (100MHz, DMSO) δ166.94,155.62,140.87,139.58,138.49,136.91,130.46,130.05,129.60,129.00,128.63,127.58,12 6.11,122.08,121.60,120.80,119.59,118.55,117.99,117.03,110.24,101.26,67.24,49.65,45.23,35.82,30.79,0.58.

[0234] Example 19-Synthesis of Compound C19:

[0235]

[0236] The synthetic route of compound C19 is as follows:

[0237]

[0238] Add 6-quinolineacetic acid (0.2736mmol, 0.051g) to a 100ml round-bottom flask, use 30ml DMF, add DIEA (0.684mmol, 0.088g), HATU (0.2736mmol, 0.104g), stir to dissolve, and finally add compound C13-1 (0.228mmol, 0.08g). After the reaction is completed, add distilled water to terminate the reaction, extract with ethyl acetate, separate the liquid to obtain the organic phase, wash with saturated ammonium chloride, saturated sodium bicarbonate, and saturated brine respectively, spin dry the solvent, and recrystallize with ethyl acetate / petroleum ether system to obtain a light yellow solid (compound C19) (0.064g, yield 54.24%).

[0239] HRMS(ESI)(M+H) + m / z 521.2551,calcd for C 32 H 33 N4O3 + 521.2547.mp182.7-184.3℃

[0240] 1 H NMR (400MHz, DMSO-d6) δ10.30(s,1H),8.87(dt,J=4.6,2.3Hz,1H),8.51(s,1H),8.36(dd,J=8.3,1.8Hz,1 H),8.03–7.95(m,1H),7.91(d,J=2.0Hz,1H),7.85–7.72(m,2H),7.53(dd,J=8.3,4.2Hz,1H),7.49–7.35(m ,5H),7.32–7.23(m,2H),7.05(s,2H),6.25(t,J=5.9Hz,1H),3.89(s,2H),3.85(ddd,J=11.5,4.5,1.8Hz, 2H), 3.26 (td, J=11.7, 2.1Hz, 2H), 3.00 (t, J=6.3Hz, 2H), 1.70–1.52 (m, 3H), 1.18 (qd, J=12.1, 4.5Hz, 2H). 13CNMR(100MHz,DMSO)δ169.38,155.59,150.65,147.28,140.81,139.97,138.40,136.16,134.85,131.78,130.12,129.51,12 9.27,128.92,128.25,128.16,127.58,126.22,122.04,121.84,118.65,118.04,117.18,67.23,45.24,43.61,35.80,30.78.

[0241] Example 20-Synthesis of Compound C20:

[0242]

[0243] The synthetic route of compound C20 is as follows:

[0244]

[0245] Add N-carbazoleacetic acid (0.2736mmol, 0.069g) to a 100ml round-bottom flask, use 30ml DMF, add DIEA (0.684mmol, 0.088g), HATU (0.2736mmol, 0.104g), stir to dissolve, and finally add compound C13-1 (0.228mmol, 0.08g). After the reaction is completed, add distilled water to terminate the reaction, extract with ethyl acetate, separate the liquid to obtain the organic phase, wash with saturated ammonium chloride, saturated sodium bicarbonate, and saturated brine respectively, spin dry the solvent, and recrystallize with ethyl acetate / petroleum ether system to obtain a white solid (compound C20) (0.086g, yield 67.7%).

[0246] HRMS(ESI)(M+H) + m / z 559.2712,calcd for C 32 H 33 N4O3 + 559.2704.mp292.2-293.4℃

[0247] 1H NMR (400MHz, DMSO-d6) δ10.76 (s, 1H), 8.90 (s, 1H), 8.18 (dt, J = 7.8, 0.9Hz, 2H), 7.87 (t,J=1.8Hz,1H),7.63(d,J=8.2Hz,2H),7.49–7.36(m,6H),7.34–7.19(m,5H),7.03( s,2H),6.52(t,J=5.9Hz,1H),5.34(s,2H),3.84(ddd,J=11.5,4.5,1.8Hz,2H),3.26( td,J=11.7,2.0Hz,2H),2.99(t,J=6.1Hz,2H),1.68–1.53(m,3H),1.26–1.10(m,2H). 13 CNMR(100MHz,DMSO)δ166.91,155.73,141.24,140.98,139.66,138.48,129.93,129.57,128.98,127.55,126.19 ,126.03,122.73,122.11,120.65,119.51,118.54,117.88,116.95,109.86,67.24,46.42,45.21,35.85,30.80.

[0248] Example 21-Synthesis of Compound C21:

[0249]

[0250] The synthetic route of compound C21 is as follows:

[0251]

[0252] C13-1a (1.536mmol, 0.4g), p-iodoaniline (1.706mmol, 0.374g), tetrabutylammonium bromide (2.303mmol, 0.742g), potassium acetate (2.559mmol, 0.251g), palladium acetate (0.0853mmol, 0.02g) were added into a pressure bottle in sequence, and 10ml of DMF was added. Under argon protection, the mixture was reacted at 80°C for 5h. After the reaction, distilled water was added to terminate the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine and dried to obtain a solid. The solid was purified by column chromatography (200-300 mesh silica gel, V(DCM):V(methanol)=10:0.1) to obtain a white solid 36 (0.24g, yield 44.44%). 2-pyridineacetic acid hydrochloride (0.2736mmol, 0.047g) was added to a 100ml round-bottom flask, and DIEA (0.684mmol, 0.088g) and HATU (0.2736mmol, 0.104g) were added with 30ml DMF, stirred to dissolve, and finally the above white solid (0.228mmol, 0.08g) was added. After the reaction was completed, distilled water was added to terminate the reaction, extracted with ethyl acetate, separated to obtain an organic phase, washed with saturated ammonium chloride, saturated sodium bicarbonate, and saturated brine, respectively, the solvent was spin-dried, and recrystallized with ethyl acetate / petroleum ether system to obtain a white solid (Compound C21) (0.070g, yield 65.42%).

[0253] HRMS(ESI)(M+H) + m / z 471.2397,calcd for C 28 H 31 N4O3 + 471.2391.mp200.3-202.5℃

[0254] 1 H NMR(400MHz,DMSO-d6)δ10.33(s,1H),8.74(s,1H),8.53–8.48(m,1H),7.76(td,J=7 .7,1.9Hz,1H),7.62(d,J=8.5Hz,2H),7.54–7.36(m,7H),7.27(ddd,J=7.6,4.9,1.2 Hz,1H),7.09–6.96(m,2H),6.41(t,J=5.9Hz,1H),3.86(m,4H),3.28(dd,J=11.8,2. 1Hz,2H),2.99(t,J=6.2Hz,2H),1.65(m,2H),1.60–1.51(m,2H),1.25–1.10(m,2H). 13CNMR(100MHz,DMSO)δ168.54,156.53,155.72,149.45,140.60,138.73,137.02,133.00,130.46,12 7.47,127.25,126.99,125.89,124.44,122.37,119.69,118.01,67.25,46.35,45.23,35.85,30.80.

[0255] Example 22-Synthesis of Compound C22:

[0256]

[0257] The synthetic route of compound C22-1 is as follows:

[0258]

[0259] Reagents and conditions of the reaction: a. 3-iodoaniline, tetrabutylammonium bromide, palladium acetate, potassium acetate, argon, 85°C; b. HATU, DIEA, DMF, 2-pyridineacetic acid; c. iron powder, ammonium chloride, water, 90°C;

[0260] 3-iodoaniline (13.5mmol, 3.348g), p-nitrostyrene (16.2mmol, 2.416g), potassium acetate (20.25mmol, 1.99g), palladium acetate (0.621mmol, 0.14g), tetrabutylammonium bromide (18.23mmol, 5.88g) were added to a 100ml pressure bottle in sequence, and 20ml dry DMF was added to dissolve. The reaction system was replaced with argon for 5 times and stirred at 80°C for 5h under argon protection. After the reaction was completed, distilled water was added to terminate the reaction, and the mixture was extracted with ethyl acetate. The upper organic phase was washed with water and saturated brine in sequence, the solvent was dried by spin drying, and then subjected to column chromatography (100-200 mesh silica gel, mobile phase pure DCM) to obtain a yellow solid (compound C22-1a).

[0261] To a 100 ml round-bottom flask, add the above yellow solid C22-1a (0.2736 mmol, 0.057 g), stir and dissolve with 30 ml dry DCM, add DIEA (0.684 mmol, 0.088 g), HATU (0.2736 mmol, 0.104 g), and finally add 2-aminomethylpyridine (0.228 mmol, 0.08 g). After the reaction, distilled water was added to terminate the reaction, extracted with ethyl acetate, separated to obtain an organic phase, washed with saturated ammonium chloride, saturated sodium bicarbonate, and saturated brine, respectively, and the solvent was dried by spin drying. The obtained solid mixture C22-1b was placed in a 100 ml round-bottom flask, iron powder (26.82 mmol, 1.5 g), ammonium chloride (16.733 mmol, 0.9 g), and 30 ml of water were added for stirring, and refluxed at 95°C for 2 h. After the reaction, it was extracted with ethyl acetate, and the residue was removed by suction filtration. The liquid was washed with saturated sodium bicarbonate, and the organic phase was washed with saturated brine after separation, and the solvent was dried by spin drying. Then, column chromatography (200-300 mesh silica gel, mobile phase V (DCM): V (methanol) = 10: 0.1) was performed to obtain a white solid (compound C22-1) (0.073 g, 81.1%).

[0262] 1 H NMR(400MHz, DMSO-d6)δ10.21(s,1H),8.51(ddd,J=4.9,1.9,1.0Hz,1H),7.81–7.72(m,2H),7.41(dt,J=7.9,1.2Hz,2H),7.31–7.2 2(m,4H),7.19(dt,J=7.8,1.4Hz,1H),6.97(d,J=16.3Hz,1H),6.85(d,J=16.3Hz,1H),6.60–6.52(m,2H),5.30(s,2H),3.86(s,2H).

[0263] The synthetic route of compound C22 is as follows:

[0264]

[0265] Compound C22-1 (0.209mmol, 0.05g) was added to a 100ml round-bottom flask, and stirred and dissolved with 10ml of anhydrous dichloromethane. CDI (0.418mmol, 0.068g) and DMAP (0.418mmol, 0.051g) were added, and the reaction was allowed to proceed overnight for 12h. After the reaction, the solvent was dried by spin drying to obtain a white powder. DMF (10ml) was added, and the mixture was stirred and dissolved. 1-(4-(aminomethyl)piperidin-1-yl)propane-1-one (0.5mmol, 0.036g) was added and the reaction was continued for 6h. After the reaction, distilled water was added to terminate the reaction, and the mixture was extracted with ethyl acetate. The organic phase was separated, washed with saturated brine, the solvent was dried by spin drying, and recrystallized with an ethyl acetate / petroleum ether system to obtain a white solid (compound C22) (0.061g, yield 79.14%).

[0266] HRMS(ESI)(M+H) + m / z 526.2813,calcd for C 31 H 36 N5O3 + 526.2813.mp183.2-184.8℃.

[0267] 1 H NMR(400MHz,DMSO-d6)δ10.28(d,J=5.1Hz,1H),8.64–8.43(m,2H),7.87–7.73 (m,2H),7.57–7.36(m,6H),7.35–7.22(m,3H),7.08(d,J=18.9Hz,2H),6.26(t, J=5.9Hz,1H),4.39(d,J=12.9Hz,1H),3.87(d,J=2.6Hz,3H),3.04–2.89(m,2H) ,2.48(s,1H),2.30(q,J=7.4Hz,2H),1.71–1.61(m,3H),0.98(t,J=7.4Hz,3H). 13 C NMR (100MHz, DMSO) δ171.43,168.73,156.50,155.57,149.47,140.78,139.99,138.37,137.04,130.13,129.50,128.88,127.58,1 26.25,124.49,122.40,121.76,118.71,118.55,118.02,117.09,46.35,45.13,44.79,41.38,36.87,30.52,29.73,26.09,10.02.

[0268] Example 23-Synthesis of Compound C23:

[0269]

[0270] The synthetic route of compound C23 is as follows:

[0271]

[0272] Compound C22-1 (0.209mmol, 0.05g) was added to a 100ml round-bottom flask, and stirred and dissolved with 10ml of anhydrous dichloromethane, and CDI (0.418mmol, 0.068g) and DMAP (0.418mmol, 0.051g) were added, and the reaction was allowed to proceed overnight for 12h. After the reaction, the solvent was dried by spin drying to obtain a white powder, and DMF (10ml) was added, and the mixture was stirred and dissolved. 4-trifluoromethoxybenzylamine (0.5mmol, 0.096g) was added and the reaction was continued for 6h. After the reaction, distilled water was added to terminate the reaction, and the mixture was extracted with ethyl acetate, and the organic phase was separated and washed with saturated brine, the solvent was dried by spin drying, and the mixture was recrystallized with an ethyl acetate / petroleum ether system to obtain a white solid (compound C23) (0.066g, yield 57.9%).

[0273] HRMS(ESI)(M+H) + m / z 547.1962,calcd for C 31 H 36 N5O3 + 547.1952.mp220.5-221.8℃.

[0274] 1 H NMR (400MHz, DMSO-d6) δ10.26(s,1H),8.73(s,1H),8.52(dd,J=4.8,1.9Hz,1H),7.83(d,J=2.2Hz,1H),7.77(td,J=7.7,1.9Hz,1H),7.52–7.40( m,7H),7.41(d,J=2.2Hz,1H),7.34(d,J=8.3Hz,2H),7.33–7.22(m,3H),7.06(s,2H),6.73(t,J=6.0Hz,1H),4.34(d,J=5.9Hz,2H),3.87(s,2H). 13C NMR (100MHz, DMSO) δ168.73,156.50,155.55,149.46,147.57,140.62,140.51,139.99,138.35,137.03,130.34,129. 50,129.37,128.85,127.57,126.37,124.48,122.39,121.77,121.45,119.31,118.58,118.23,117.11,46.35,42.52.

[0275] Example 24-Synthesis of Compound C24:

[0276]

[0277] The synthetic route of compound C24 is as follows:

[0278]

[0279] 3-iodobenzoic acid (13.5mmol, 3.348g), p-nitrostyrene (16.2mmol, 2.416g), potassium acetate (20.25mmol, 1.99g), palladium acetate (0.621mmol, 0.14g), tetrabutylammonium bromide (18.23mmol, 5.88g) were added to a 100ml pressure bottle in sequence, and 20ml dry DMF was added to dissolve. The reaction system was replaced with argon for 5 times and stirred at 80°C for 5h under argon protection. After the reaction was completed, distilled water was added to terminate the reaction, and the mixture was extracted with ethyl acetate. The upper organic phase was washed with water and saturated brine in sequence, the solvent was dried by spin drying, and then subjected to column chromatography (100-200 mesh silica gel, mobile phase pure DCM) to obtain red solid C24-1a. To a 100 ml round-bottom flask, C24-1a (0.2736 mmol, 0.048 g) was added, dissolved by stirring with 30 ml of dry DCM, and DIEA (0.684 mmol, 0.088 g), HATU (0.2736 mmol, 0.104 g) were added, and finally 2-aminomethylpyridine (0.228 mmol, 0.08 g) was added. After the reaction, distilled water was added to terminate the reaction, extracted with ethyl acetate, separated to obtain an organic phase, washed with saturated ammonium chloride, saturated sodium bicarbonate, and saturated brine, respectively, and the solvent was dried by spin drying. The obtained solid mixture C24-1b was placed in a 100 ml round-bottom flask, iron powder (26.82 mmol, 1.5 g), ammonium chloride (16.733 mmol, 0.9 g), and 30 ml of water were added for stirring, and refluxed at 95 ° C for 2 h. After the reaction, it was extracted with ethyl acetate, and the filter residue was removed by suction. The liquid was washed with saturated sodium bicarbonate, and the organic phase was washed with saturated brine after separation, and the solvent was dried by spin drying. Then, column chromatography (200-300 mesh silica gel, mobile phase V (DCM): V (methanol) = 10:0.1) was performed to obtain a white solid C24-1c. Compound C24-1c (0.209mmol, 0.05g) was added to a 100ml round-bottom flask, and stirred and dissolved with 10ml of anhydrous dichloromethane. CDI (0.418mmol, 0.068g) and DMAP (0.418mmol, 0.051g) were added, and the reaction was allowed to proceed overnight for 12h. After the reaction, the solvent was dried to obtain a white powder. DMF (10ml) was added, and the mixture was stirred and dissolved. 4-aminomethyltetrahydropyran (0.5mmol, 0.036g) was added and the reaction was continued for 6h. After the reaction, distilled water was added to terminate the reaction, and the mixture was extracted with ethyl acetate. The organic phase was separated, washed with saturated brine, the solvent was dried to obtain a white solid (compound C24) (0.054g, yield 77.14%).

[0280] HRMS(ESI)(M+H) + m / z 471.2402,calcd for C 28 H31 N4O3 + 471.2391.mp212.4-214.1℃

[0281] 1 H NMR(400MHz, DMSO-d6)δ9.47–9.43(s,1H),9.30(t,J=6.0Hz,1H),8.52(dt,J=4.9,1.4Hz,1H),8 .18(s,1H),7.84–7.67(m,3H),7.52–7.41(m,5H),7.36(d,J=8.0Hz,1H),7.33–7.25(m,2H),7.1 4(d,J=16.4Hz,1H),6.89(d,J=6.3Hz,1H),4.60(d,J=5.9Hz,2H),3.84(ddd,J=11.3,4.6,1.7Hz ,2H),3.26(td,J=11.7,1.9Hz,2H),2.99(t,J=6.0Hz,2H),1.69–1.55(m,3H),1.25–1.10(m,2H). 13 C NMR (100MHz, DMSO) δ166.82,159.34,155.93,149.26,141.31,138.16,137.22,135.04,129.85,129.77, 129.56,129.19,127.58,126.42,125.38,125.07,122.54,121.40,117.75,67.26,45.17,35.92,30.83.

[0282] Example 25-Synthesis of Compound C25:

[0283]

[0284] The synthetic route of compound C25-1 is as follows:

[0285]

[0286] C1-1a (6.75mmol, 1g), m-iodoaniline (6.075mmol, 1.33g), tetrabutylammonium bromide (9.113mmol, 2.938g), potassium acetate (10.125mmol, 1g), palladium acetate (0.3375mmol, 0.076g) were added into a pressure bottle in sequence, and 10ml of DMF was added. Under argon protection, the mixture was reacted at 80°C for 5h. After the reaction, distilled water was added to terminate the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine and dried to obtain a solid. The solid was purified by column chromatography (200-300 mesh silica gel, V (petroleum ether): V (ethyl acetate) = 10:1) to obtain a yellow solid (compound C25-1) (0.9g, yield 63.7%).

[0287] The synthetic route of compound C25 is as follows:

[0288]

[0289] Compound C25-1 (0.209mmol, 0.05g) was added to a 100ml round-bottom flask, and stirred and dissolved with 10ml of anhydrous dichloromethane. CDI (0.418mmol, 0.068g) and DMAP (0.418mmol, 0.051g) were added, and the reaction was allowed to proceed overnight for 12h. After the reaction, the solvent was dried by spin drying to obtain a white powder. DMF (10ml) was added, and the mixture was stirred and dissolved. Isobutylamine (0.5mmol, 0.036g) was added and the reaction was continued for 6h. After the reaction, distilled water was added to terminate the reaction, and the mixture was extracted with ethyl acetate. The organic phase was separated, washed with saturated brine, the solvent was dried by spin drying, and recrystallized with an ethyl acetate / petroleum ether system to obtain a white solid (compound C25) (0.054g, yield 77.14%).

[0290] HRMS(ESI)(M+H) + m / z 339.1702,calcd for C 20 H 23 N2O3 + 339.1703.mp219.8-221.6℃. 1H NMR (400MHz, DMSO-d6) δ8.40(s,1H),7.59(t,J=1.9Hz,1H),7.28(d,J=1.7Hz,1 H),7.24(dt,J=8.1,1.6Hz,1H),7.20(t,J=7.7Hz,1H),7.10(dt,J=7.6,1.6Hz,1 H),7.03(d,J=12.6Hz,3H),6.90(d,J=8.0Hz,1H),6.20(t,J=5.9Hz,1H),6.03(s ,2H),2.93(t,J=6.3Hz,2H),1.77–1.63(m,J=6.7Hz,1H),0.88(d,J=6.6Hz,6H). 13 C NMR(101MHz,DMSO)δ155.76,148.36,147.43,141.40,138.08,132.04,129.37,128.40, 127.50,122.08,119.58,117.29,115.85,108.85,105.89,101.54,47.01,28.96,20.48.

[0291] Example 26-Synthesis of Compound C26:

[0292]

[0293] The synthetic route of compound C26 is as follows:

[0294]

[0295] Compound C25-1 (0.209mmol, 0.05g) was added to a 100ml round-bottom flask, and stirred and dissolved with 10ml of anhydrous dichloromethane, and CDI (0.418mmol, 0.068g) and DMAP (0.418mmol, 0.051g) were added, and the reaction was continued overnight for 12h. After the reaction, the solvent was dried to obtain a white powder, and DMF (10ml) was added, and stirred and dissolved. 4-aminomethyltetrahydropyran (0.5mmol, 0.06g) was added and reacted for 6h. After the reaction, distilled water was added to terminate the reaction, and the mixture was extracted with ethyl acetate, and the organic phase was separated, washed with saturated brine, the solvent was dried, and recrystallized with ethyl acetate / petroleum ether system to obtain a white solid (compound C26) (0.066g, yield 83.1%).

[0296] HRMS(ESI)(M+H) + m / z 381.1803,calcd for C 22 H 25 N2O4 +381.1809.mp207.0-208.6℃

[0297] 1 H NMR(400MHz, DMSO-D6)δ:8.42(s,1H),7.59(t,J=1.8Hz,1H),7.29(d,J=1.6Hz,1H),7.27–7.16( m,2H),7.10(dt,J=7.3,1.6Hz,1H),7.04(d,J=11.7Hz,3H),6.91(d,J=8.0Hz,1H),6.24(t,J=5.9 Hz,1H),6.04(s,2H),3.85(m,J=11.5,4.6,1.9Hz,2H),3.27(td,J=11.7,2.1Hz,2H),3.00(t,J=6 .2Hz,2H),1.66(m,J=10.6,6.6,3.2Hz,1H),1.56(m,J=12.9,4.0,1.9Hz,2H),1.25–1.11(m,2H). 13 C NMR(100MHz,DMSO)δ:155.78,148.36,147.43,141.34,138.07,132.02,129.38,128.40,127 .47,122.11,119.61,117.31,115.87,108.86,105.87,101.54,67.25,45.25,35.84,30.80.

[0298] Example 27-Synthesis of Compound C27:

[0299]

[0300] The synthetic route of compound C27 is as follows:

[0301]

[0302] The specific steps include: adding compound C25-1 (0.209mmol, 0.05g) to a 100ml round-bottom flask, stirring and dissolving with 10ml of anhydrous dichloromethane, adding CDI (0.418mmol, 0.068g), DMAP (0.418mmol, 0.051g), reacting overnight for 12h, spinning the solvent to dryness after the reaction to obtain a white powder, adding DMF (10ml), stirring and dissolving, adding isopentylamine (0.5mmol, 0.044g), and reacting for 6h. After the reaction, adding distilled water to terminate the reaction, extracting with ethyl acetate, separating the organic phase, washing with saturated brine, spinning the solvent to dryness, and recrystallizing with ethyl acetate / petroleum ether system to obtain a white solid (compound C27) (0.053g, yield 72.0%).

[0303] HRMS(ESI)(M+H) + m / z 353.1844,calcd for C 21 H 25 N2O3 + 353.1860.mp208.1-212.2℃

[0304] 1 H NMR(400MHz, DMSO-D6)δ:8.46(s,1H),7.59(t,J=1.9Hz,1H),7.29(d,J=1.8H z,1H),7.26–7.16(m,2H),7.09(dt,J=7.3,1.6Hz,1H),7.03(d,J=12.1Hz,3H ),6.91(d,J=8.0Hz,1H),6.15(t,J=5.6Hz,1H),6.03(s,2H),3.16–3.06(m,2 H),1.69–1.54(m,J=6.7Hz,1H),1.33(q,J=7.0Hz,2H),0.90(d,J=6.6Hz,6H). 13 C NMR (100MHz, DMSO) δ155.70,148.35,147.42,141.42,138.04,132.03,129.35,128.37, 127.50,122.10,119.54,117.29,115.86,108.85,105.87,101.54,37.71,25.61,22.88.

[0305] Example 28-Synthesis of Compound C28:

[0306]

[0307] The synthetic route of compound C28-1a is as follows:

[0308]

[0309] 3,5-dimethoxybenzaldehyde (6.02mmol, 1g), methyltriphenylphosphine bromide (9.03mmol, 3.226g), potassium tert-butoxide (10.03mmol, 1.126g) were added to a 100ml round-bottom flask, and then 30ml of tetrahydrofuran was added to stir and dissolve, and the mixture was reacted for 2h under argon protection in an ice bath. After the reaction, the mixture was quenched with a saturated ammonium chloride solution, extracted with dichloromethane, and the organic phase was washed with saturated brine after separation. The solvent was dried to obtain an orange liquid. The mixture was then subjected to column chromatography (100-200 mesh silica gel, mobile phase V (petroleum ether): V (ethyl acetate) = 15:1) to obtain a colorless transparent liquid (compound C28-1a) (0.8g, yield 80.97%).

[0310] The synthetic route of compound C28-1 is as follows:

[0311]

[0312] C28-1a (5.483mmol, 0.9g), m-iodoaniline (4.935mmol, 1.081g), tetrabutylammonium bromide (7.402mmol, 2.386g), potassium acetate (8.225mmol, 0.807g), palladium acetate (0.274mmol, 0.062g) were added into a pressure bottle in sequence, and 10ml of DMF was added. Under argon protection, the mixture was reacted at 80°C for 5h. After the reaction, distilled water was added to terminate the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine and dried to obtain a solid. The solid was purified by column chromatography (200-300 mesh silica gel, V (petroleum ether): V (ethyl acetate) = 10:1) to obtain a brown viscous liquid (compound C28-1) (0.68g, yield 53.97%). 1 H NMR(400MHz, DMSO-D6)δ7.11(d,J=16.4Hz,2H),7.07–6.95(m,2H),6.80–6.72(m,2H) ,6.50(ddd,J=7.9,2.3,1.0Hz,2H),6.40(t,J=2.2Hz,2H),5.07(s,2H),3.78(s,6H).

[0313] The synthetic route of compound C28 is as follows:

[0314]

[0315] Compound C28-1 (0.2mmol, 0.05g) was added to a 100ml round-bottom flask, and stirred and dissolved with 10ml of anhydrous dichloromethane. CDI (0.418mmol, 0.068g) and DMAP (0.418mmol, 0.051g) were added, and the reaction was allowed to proceed overnight for 12h. After the reaction, the solvent was dried to obtain a white powder. DMF (10ml) was added, and the mixture was stirred and dissolved. Isobutylamine (0.5mmol, 0.037g) was added and the reaction was continued for 6h. After the reaction, distilled water was added to terminate the reaction, and the mixture was extracted with ethyl acetate. The organic phase was separated, washed with saturated brine, the solvent was dried to obtain a white solid (compound C28) (0.053g, yield 74.76%).

[0316] HRMS(ESI)(M+H) + m / z 355.2022,calcd for C 21 H 27 N2O3 + 355.2106.mp160.1-163.5℃.

[0317] 1 H NMR(400MHz,)δ8.74(s,1H),7.68(t,J=1.9Hz,1H),7.30–7.17(m,3H),7.17–6.99(m,2H),6.78 (d,J=2.2Hz,2H),6.49–6.38(m,2H),3.78(s,6H),2.93(t,J=6.2Hz,2H),0.88(d,J=6.7Hz,6H). 13 CNMR(100MHz,DMSO)δ165.88,160.64,146.31,144.27,142.46,134.55,134.14 ,133.35,124.49,122.33,120.71,109.65,105.22,60.44,51.72,33.74,25.26.

[0318] Example 29-Synthesis of Compound C29:

[0319]

[0320] The synthetic route of compound C29 is as follows:

[0321]

[0322] Compound C28-1 (0.2mmol, 0.05g) was added to a 100ml round-bottom flask, and stirred and dissolved with 10ml of anhydrous dichloromethane. CDI (0.4mmol, 0.065g) and DMAP (0.4mmol, 0.05g) were added, and the reaction was continued overnight for 12h. After the reaction, the solvent was dried to obtain a white powder, and DMF (10ml) was added, and stirred and dissolved. 4-aminomethyltetrahydropyran (0.5mmol, 0.06g) was added and the reaction was continued for 6h. After the reaction, distilled water was added to terminate the reaction, and the mixture was extracted with ethyl acetate, and the organic phase was separated and washed with saturated brine, the solvent was dried to obtain a white solid (compound C29) (0.062g, yield 78.18%).

[0323] HRMS(ESI)(M+H) + m / z 397.2123,calcd for C 23 H 29 N2O4 + 397.2122.mp159.4-162.8℃

[0324] 1 H NMR (400MHz, DMSO-d6) δ8.43(s,1H),7.68(s,1H),7.26–7.17(m,3H),7.14(t,J=4.7Hz,1H ),7.06(d,J=16.4Hz,1H),6.78(d,J=2.2Hz,2H),6.42(d,J=2.3Hz,1H),6.26(t,J=5.9Hz, 1H),3.85(ddd,J=11.3,4.7,1.8Hz,2H),3.78(s,6H),3.29–3.21(m,2H),3.01(t,J=6.2Hz ,2H),1.66(dqd,J=10.6,6.7,2.7Hz,1H),1.60–1.52(m,2H),1.18(qd,J=12.1,4.5Hz,2H). 13 C NMR (100MHz, DMSO) δ161.15,155.80,141.39,139.51,137.77,129.77,129.42,128 .67,119.91,117.72,116.14,104.95,100.46,67.25,55.70,45.26,35.85,30.80.

[0325] The technical effects and advantages of the present invention are verified below by applying the stilbene compound of the present invention to inhibit soluble epoxide hydrolase.

[0326] Specifically, the verification method of the stilbene compound in the embodiment of the present invention in inhibiting soluble epoxide hydrolase is to incubate the stilbene compound in the embodiment of the present invention with a sample containing soluble epoxide hydrolase and the endogenous hydrolysis substrate 14,15-EET of the hydrolase, and detect the content of the hydrolysis product 14,15-DHET, and use its relative production amount to reflect the inhibitory effect of the compound on the soluble epoxide hydrolase. The specific steps are as follows:

[0327] 1:10 (mg / μL) pre-cooled PBS (phosphate buffer saline) (pH=7.4) was added to the collected brain tissue samples of adult (weight 25-30g) male C57BL / 6J mice, and the samples were disrupted with a homogenizer (Bertin Precellys 24-Dual, Bertin Technologies) for 30 seconds (4°C, 4500rpm), and then centrifuged at 9000g for 15 minutes at 4°C in a small low-temperature centrifuge, and the supernatant was diluted 20 times. In an EP (eppendorf) tube, 160 μL PBS, 20 μL tissue fluid diluted 20 times, and 2 μL of a compound with a concentration gradient of 10000, 5000, 1000, 500, 100, 50, 10, 1, 0.1, and 0.01 μg / mL (final concentration 100, 50, 10, 5, 1, 0.5, 0.1, 0.01, 0.001, and 0.0001 μg / mL) were added and incubated at room temperature for 15 minutes. The negative control was PBS without tissue diluent but with solvent added (180 μL PBS plus 2 μL DMSO), and the positive control was the same concentration sample diluent plus an equal volume of solvent (160 μL PBS plus 20 μL tissue fluid diluted 20 times plus 2 μL DMSO). After the room temperature incubation, all samples were placed on ice, and the enzyme reaction substrate (10 μL 14,15-EET (10 μg / mL)) was quickly added and mixed, and then incubated in a shaker at 37°C. After the incubation, the samples were placed on ice and 10 μL 800 nM t-TUCB (No. 6757, Tocris Bioscience, UK, Bristol) was quickly added to terminate the reaction, and the final volume of the reaction system was 200 μL. In order to detect the enzyme activity, the concentration of the reaction product 14,15-DHET was detected using high performance liquid chromatography-mass spectrometry. The pretreatment is as follows: 1. Add an equal volume (200 μL) of a mixture of methanol and acetonitrile (50 / 50, v / v) to the sample, the mixture contains 0.4% acetic acid and internal standards 11,12-EET-d11 and 11,12-DHET-d11 (20 ng / mL); 2. Mix with a homogenizer at 4500 rpm for 10 minutes and place at -20°C for 2 hours to precipitate the protein; 3. After the sample is centrifuged for 10 minutes (14000 rpm), the supernatant is collected for UPLC-MS / MS analysis. The relative enzyme activity of sEH is calculated by comparing the amount of 14,15-DHET produced at a certain concentration of the test compound with that without an inhibitor, and the inhibition rate is 100% minus the relative enzyme activity.

[0328] The inhibition rate %@5 μg / mL is the inhibition rate of the compound on sEH at 5 μg / mL.

[0329] Among them, the positive control compounds are TPPU and EC5026.

[0330] The verification results of the diphenylethylene compounds prepared by the present invention are shown in Table 1:

[0331] Table 1: Inhibitory activity of the stilbene compounds prepared in this application on soluble epoxide hydrolases:

[0332]

[0333]

[0334]

[0335] The results of the positive control compounds are shown in Table 2:

[0336] Table 2. Inhibitory activity of positive control compounds on soluble epoxidase under the same conditions

[0337]

[0338] Table 3. Inhibitory activity of weakly active control compounds on soluble epoxide enzymes under the same conditions

[0339]

[0340]

[0341] As can be seen from Tables 1 and 2, the IC50 of the positive control compounds TPPU and EC5026 against soluble epoxide hydrolase (sEH) are 44 nm or 19 nm, respectively, but the stilbene skeleton compounds prepared by the present invention have comparable or better sEH inhibitory activity (especially when the urea group is in the para position of the stilbene, that is, the IC50 of all compounds in the general formula II is less than 9 nm), indicating that the stilbene skeleton can enhance the interaction with sEH. WS-54 and WS-82 are known compounds, but the inhibitory activity of sEH has not been reported. As can be seen from Table 3, the inhibition rates of the control compounds WS-54 and WS-82 against sEH inhibition are reduced to about 12% and 91%, respectively, and the IC50 of the control compound WS-82 reaches 644 nm, indicating that the urea group with NH on both sides in the general formula is one of the characteristic structures for inhibiting sEH. Compared with the results of the inhibition rate and IC50 of the control compounds XWS-50 and XWS-65 on sEH, it is shown that a methylene group must be retained between the R4 group and the NH group of the urea group in the general formula to obtain a better inhibition rate and a lower IC50 for sEH. Therefore, the present invention introduces the diphenylethylene described in the general formula I to obtain a new type of sEH inhibitory compound, which has a good inhibition rate for sEH, and the lowest IC50 is about 1nm.

[0342] In summary, the main skeleton of the diphenylethylene compound prepared by the present invention is diphenylethylene with a urea group. By expanding different substituents on the urea side and growing different groups on the diphenylethylene side skeleton containing the dominant structure of natural products, a series of highly active diphenylethylene compounds are obtained, and the IC50 value of the compound for inhibiting sEH by the highly active diphenylethylene compounds reaches the nanomolar level, indicating that the diphenylethylene compounds have a high efficiency inhibitory effect on sEH, thereby achieving the inhibitory effect on inflammation by inhibiting sEH, and have good application prospects.

[0343] The above is a detailed introduction to a diphenylethylene compound, a pharmaceutical composition and its application provided by the present invention. Specific examples are used herein to illustrate the principle and implementation mode of the present invention. The description of the above embodiments is only used to help understand the method of the present invention and its core idea. At the same time, for a person skilled in the art, according to the idea of ​​the present invention, there will be changes in the specific implementation mode and the scope of application. In summary, the content of this specification should not be understood as a limitation on the present invention.

Claims

1. A stilbene compound, characterized in that: The diphenylethylene compound has the following general structural formula I: In the formula I, R 1 , R 2 The groups are combined to form methylenedioxy, or independently selected from methoxy, hydroxyl, hydrogen and any one of the following R1-R9 groups, the structural formula of the R1-R9 groups is as follows: R 3 Any one group selected from methoxy and hydrogen; R 4 Selected from C 1-4 Alkyl, tetrahydropyranyl, N-(C 1-4 Any one of 4-(4-(6-(4-(6-(4-(2 ... X and Y are each independently selected from any one of methyl, hydrogen, fluorine and chlorine.

2. The compound according to claim 1, characterized in that When the urea group of the compound is located at the para position of the diphenylethylene skeleton, the general structural formula is as shown in Formula II: In the formula II, R 1 , R 2 , R 3 , R 4 , X, and Y are the same as those defined in claim 1.

3. The compound according to claim 1, characterized in that When the urea group of the compound is located at the meta position of the diphenylethylene skeleton, and X=Y=H, the general structural formula is as shown in Formula Ш: In the formula, R 1 , R 2 , R 3 , R 4 , X, and Y are the same as those defined in claim 1.

4. The compound according to claim 1, characterized in that The R 1 , R 2 Combined to form methylenedioxy, or independently selected from methoxy, hydrogen, any one of R1-R9; R 3 Any one group selected from methoxy and hydrogen; R 4 Any one group selected from isopropyl, isobutyl, tetrahydropyranyl, N-acetylpiperidinyl, N-propionylpiperidinyl and 4-trifluoromethoxyphenyl.

5. The compound according to claim 1, characterized in that X of the compound is selected as hydrogen, and Y of the compound is selected as hydrogen.

6. The compound according to claim 1, characterized in that The R 1 , R 2 Select methylenedioxy, R 3 Select hydrogen, R 4 is tetrahydropyranyl, X is hydrogen, and Y is hydrogen; or The R 1 Select methoxy, R 2 Select hydrogen, R 3 is methoxy, R 4 is tetrahydropyranyl, X is hydrogen, and Y is hydrogen; or The R 1 Select any one of the groups R1-R9, R 2 Select hydrogen, R 3 Select hydrogen, R 4 is tetrahydropyranyl, X is hydrogen, and Y is hydrogen; or The R 2 Select any one of the groups R1-R9, R 1 Select hydrogen, R 3 Select hydrogen, R 4 Select tetrahydropyranyl, X is hydrogen, and Y is hydrogen.

7. The compound according to claim 6, characterized in that The structural formula of the compound is:

8. The compound according to claim 1, characterized in that The specific structural formula of the compound is:

9. A pharmaceutical composition of a stilbene compound, characterized in that: The distilbene compound pharmaceutical composition comprises at least one of the compound according to any one of claims 1 to 8, its isotope label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof.

10. Use of the stilbene compound according to any one of claims 1 to 8, or the stilbene compound pharmaceutical composition according to claim 9, in the preparation of anti-inflammatory drugs.

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