Biaryl hydrocarbon compounds and their preparation methods and applications

Abstract

The present invention belongs to the field of pharmaceutical chemistry and discloses biaryl hydrocarbon compounds, their preparation methods, and applications. The biaryl hydrocarbon compounds have the structure represented by the general formula (I): #imgabs0#, wherein the aromatic ring is any one of an arbitrarily substituted aromatic ring, an arbitrarily substituted five-membered or six-membered aromatic heterocycle, and an arbitrarily substituted naphthalene ring. Pharmacological experiments on the biaryl hydrocarbon compounds provided by the present invention have shown that the compounds inhibit tubulin polymerization, arrest cells in the G2 / M phase, and induce apoptosis in tumor cells. These compounds are useful in treating malignant tumors caused by an imbalance in microtubule homeostasis.

Description

Technical Field

[0001] The present invention belongs to the field of medicinal chemistry and relates to a class of biaryl hydrocarbon compounds as microtubule polymerization inhibitors, in particular to biaryl hydrocarbon compounds and preparation methods and applications thereof. Background Art

[0002] Microtubules perform a variety of biological functions in eukaryotic cells, such as cell division and proliferation, cytoskeleton maintenance, and cell motility. In tumor cells, microtubules are abnormally activated, so disrupting their dynamic equilibrium has become a therapeutic approach. Based on their mode of action, microtubule-targeted drugs can be divided into microtubule polymerization promoters (such as paclitaxel) and microtubule polymerization inhibitors (such as vincristine). Currently, microtubule-targeted drugs are marketed, but they face significant toxic side effects, poor physicochemical properties, and the development of multidrug resistance during clinical use. The colchicine site is located at the junction of microtubule α and β proteins. Colchicine site inhibitors are microtubule polymerization inhibitors that interfere with the dynamic polymerization of microtubules and inhibit cell mitosis. They can also act as vascular disruptors, destroying blood vessels in tumors and inhibiting the supply of oxygen and nutrients to the tumor. At the same time, colchicine site inhibitors still have inhibitory activity against drug-resistant tumor cells. Currently, various structural types such as colchicine derivatives, CA-4 derivatives, chalcone analogs, etc. have been studied and reported as colchicine site inhibitors (Curr. Med. Chem., 2020, 27(40): 6787-6814). However, so far, no colchicine site inhibitors have been marketed. Therefore, the development of colchicine site inhibitors has important practical significance for tumor treatment, especially the treatment of drug-resistant tumors. Summary of the Invention

[0003] The invention discloses a class of biaryl hydrocarbon compounds, and provides a specific preparation method of the compounds and the pharmaceutical application of the compounds as microtubule polymerization inhibitors.

[0004] To achieve the above object, the technical solution of the present invention is implemented as follows:

[0005] Biaryl hydrocarbon compounds, the general structural formula of which is shown in formula (I):

[0006]

[0007] The aromatic ring is any one of an optionally substituted aromatic ring, an optionally substituted five-membered or six-membered aromatic heterocycle, or an optionally substituted naphthalene ring.

[0008] Furthermore, the aromatic ring in the biaryl hydrocarbon compound is Any one of the following; where R 1is any one of H, F, Cl, Br, CH3, CH2CH3, OH, OCH3, CF3, OCF3, CN, CH2OH, NH2, NHCH3 and N(CH3)2, R 1 It is any one of mono-, di-, and tri-substituted.

[0009] Furthermore, the biaryl hydrocarbon compound is a compound having the following structure:

[0010]

[0011] The preparation method of the above-mentioned biaryl hydrocarbon compound is as follows:

[0012]

[0013] The aromatic ring is any one of an optionally substituted aromatic ring, an optionally substituted five-membered or six-membered aromatic heterocycle, and an optionally substituted naphthalene ring.

[0014] The specific preparation steps are:

[0015] (1) Preparation of Compound III

[0016] Compound II is used as a starting material to undergo a nucleophilic reaction with glycine methyl ester hydrochloride under alkaline conditions (potassium carbonate or cesium carbonate) and in acetonitrile or N,N-dimethylformamide (DMF) as a solvent to generate compound III;

[0017] (2) Preparation of Compound IV

[0018] Compound III undergoes a reduction reaction with SnCl2 and further undergoes a cyclization reaction to generate compound IV;

[0019] (3) Preparation of Compound V

[0020] Compound IV is acylated with 3,4,5-trimethoxybenzoyl chloride in the presence of DMAP in anhydrous tetrahydrofuran as solvent to produce compound V.

[0021] (4) Preparation of Compound Ⅰ

[0022] Compound V and aromatic boronic acid VI are reacted under alkaline conditions (sodium carbonate, potassium carbonate or cesium carbonate) and a palladium catalyst (PdCl2(dppf), Pd(OAc)2 or Pd(PPh3)4) in 1,4-dioxane, tetrahydrofuran or ethylene glycol dimethyl ether as solvent to undergo Suzuki reaction to obtain the target product I.

[0023] In the step (4), the aromatic boronic acid VI is any one of 2-thiopheneboronic acid, 3-thiopheneboronic acid, furan-2-boronic acid, furan-3-boronic acid, 2-hydroxyphenylboronic acid, 3-hydroxyphenylboronic acid, 4-hydroxyphenylboronic acid, 2-hydroxymethylphenylboronic acid, 3-hydroxymethylphenylboronic acid, 4-hydroxymethylphenylboronic acid, 2-chlorophenylboronic acid, 3-chlorophenylboronic acid, 4-chlorophenylboronic acid, 2-methylphenylboronic acid, 3-methylphenylboronic acid, 4-methylphenylboronic acid, 2-cyanophenylboronic acid, 3-cyanophenylboronic acid, 4-cyanophenylboronic acid, 2-fluorophenylboronic acid, 3-fluorophenylboronic acid, 4-fluorophenylboronic acid, 2-methoxyphenylboronic acid, 3-methoxyphenylboronic acid, 4-methoxyphenylboronic acid, phenylboronic acid, 2-naphthaleneboronic acid, 1-naphthaleneboronic acid and 3,5-dichlorophenylboronic acid.

[0024] A pharmaceutical composition comprising the aforementioned biaryl hydrocarbon compound and a pharmaceutically acceptable carrier. The compound can be added with a pharmaceutically acceptable carrier to form common pharmaceutical preparations such as tablets, capsules, syrups, suspensions, and injections. Common pharmaceutical excipients such as flavorings, sweeteners, liquid or solid fillers, or diluents can also be added.

[0025] When the aromatic ring is Any one of R 1 is any one of H, F, Cl, Br, CH3, CH2CH3, OH, OCH3, CF3, OCF3, CN, CH2OH, NH2, NHCH3 and N(CH3)2, R 1 The biaryl hydrocarbon compounds are not limited to the compounds prepared above. Other biaryl hydrocarbon compounds not listed in the present invention can also act on the colchicine site based on the bioisosteric principle in medicinal chemistry and molecular docking results, and are used as microtubule polymerization inhibitors.

[0026] The use of the compound of formula (I) and its stereoisomers, hydrates, solvates, or crystals in the preparation of microtubule polymerization inhibitors is also within the scope of the present invention. The microtubule polymerization inhibitors are used to prepare drugs for treating malignant tumors, such as cervical cancer, liver cancer, lung cancer, and breast cancer.

[0027] The beneficial effects produced by the present invention are:

[0028] (1) The present invention discloses a biaryl hydrocarbon compound represented by the general formula (I). Pharmacological experiments show that the compound (I) of the present invention has an inhibitory effect on tumor cell proliferation by inhibiting microtubule polymerization, interfering with cell mitosis, and arresting cells in the G2 / M phase. Specifically, some compounds exhibit nanomolar inhibitory effects, among which compounds I-2, I-4, and I-5 exhibit strong inhibitory activity against two types of tumor cells (HeLa and HepG2), slightly weaker than the positive drugs Colchicine and CA-4, and better than DOX.

[0029] (2) The compounds disclosed in the present invention can be used to treat malignant tumors caused by excessive microtubule activation.

[0030] (3) The present invention also discloses a method for preparing the biaryl hydrocarbon compound. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0032] Figure 1 The inhibitory effect of compound Ⅰ-2 on microtubule polymerization.

[0033] Figure 2 Figure 1 is the blocking effect of compound Ⅰ-2 on the cell cycle. A is a flow cytometric graph of the effects of different concentrations of Ⅰ-2 on the cell cycle. B is a statistical graph of the effects of Ⅰ-2 on the cell cycle.

[0034] Figure 3 The docking diagram of compound I-2 and microtubules (Figure A); the superposition diagram of compound I-2 and colchicine (Figure B). DETAILED DESCRIPTION

[0035] The following will clearly and completely describe the technical solutions of the present invention in conjunction with the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0036] Example 1

[0037] The synthesis of 7-(thiophen-2-yl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-1) in this example is as follows:

[0038]

[0039] (1) Synthesis of (4-bromo-2-nitrophenyl)glycine methyl ester (Ⅲ-1)

[0040] 4-Bromo-1-fluoro-2-nitrobenzene (II-1) (5.00 g, 22.70 mmol), glycine methyl ester hydrochloride (3.14 g, 24.97 mmol), and potassium carbonate (6.27 g, 45.40 mmol) were added sequentially to a 100 mL round-bottom flask. Dissolved in 50 mL of anhydrous acetonitrile, the mixture was stirred at room temperature for 24 hours, and the reaction was monitored by TLC. After completion of the reaction, the mixture was evaporated under reduced pressure and purified by column chromatography (PE:EA = 2:1) to yield 5.80 g of a brownish-red solid in 88.4% yield. Melting point: 132.5-133°C. 1 H NMR (300MHz, CDCl3) δ8.38 (s, 1H), 8.36 (d, J = 2.4Hz, 1H), 7.53 (dd, J = 9.0, 2.4Hz, 1H), 6.61 (d, J = 9.0Hz, 1H), 4.10 (d, J = 5.3Hz, 1H), 3.83 (s, 3H).

[0041] (2) Synthesis of 7-bromo-3,4-dihydroquinoxaline-2(1H)-one (Ⅳ-1)

[0042] Intermediate III-1 (2.00 g, 6.92 mmol) and stannous chloride (7.80 g, 41.52 mmol) were added sequentially to a 100 mL round-bottom flask and dissolved in 30 mL of ethyl acetate. Once dissolved, 1 mL of water was added and the mixture was heated to 50°C for 24 hours. The reaction was monitored by TLC. Upon completion, the mixture was cooled to room temperature, washed with water, dried over anhydrous sodium sulfate, and evaporated under reduced pressure. Purification by column chromatography (PE:EA = 2:1) afforded 1.3 g of a light brown solid with a yield of 82.7%. Melting point: 255.7-256.9°C. 1 H NMR (300MHz, DMSO-d6) δ10.34(s,1H),6.96-6.82(m,2H),6.60(d,J=8.3Hz,1H),6.15(s,1H),3.74(d,J=1.6Hz,2H).

[0043] (3) Synthesis of 7-bromo-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (V-1)

[0044] Intermediate IV-1 (1.00 g, 4.40 mmol) and DMAP (108 mg, 0.88 mmol) were dissolved in 15 mL of anhydrous tetrahydrofuran. Under nitrogen, a solution of 3,4,5-trimethoxybenzoyl chloride (1.50 g, 6.60 mmol) in anhydrous tetrahydrofuran was added dropwise under an ice bath. The mixture was allowed to react at room temperature for 24 hours. The reaction was monitored by TLC. Upon completion, the product was evaporated under reduced pressure and purified by column chromatography (DCM:MeOH = 30:1) to afford 912 mg of a brownish-yellow solid in a 49.2% yield. Melting point: 213.9-214.3°C. 1 H NMR (300MHz, DMSO-d6) δ10.89(s,1H),7.19(d,J=2.0Hz,1H),7.04(dd,J=8.6,1. 7Hz,1H),6.89-6.78(m,1H),6.71(s,2H),4.39(s,2H),3.71(s,3H),3.69(s,6H). 13 C NMR(75MHz,DMSO-d6)δ168.52,167.04,153.01,139.98,133.28,129.59,126.74,1 26.47,124.65,118.84,117.80,106.69,60.64,56.41,48.37.HRMS(ESI)cald.for C 18 H 17 N2O5BrNa + [M+Na] + :443.02131,found:443.02150.

[0045] (4) Synthesis of 7-(thiophen-2-yl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-1)

[0046] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 2-thiopheneboronic acid (73.7 mg, 0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then stirred at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. Purification by column chromatography (DCM:MeOH = 50:1) afforded 66 mg of a pale white solid in a 32.4% yield. Melting point: 207.6-208.9°C. 1H NMR(300MHz, CDCl3)δ9.62(s,1H),7.25-7.20(m,2H),7.17-7.13(m,1H),7.04-6.98 (m,2H),6.69(d,J=7.8Hz,1H),6.61(s,2H),4.57(s,2H),3.80(s,3H),3.64(s,6H). 13 C NMR (75MHz, CDCl3) δ168.73,168.70,153.00,142.46,140.61,132.62,130.25,128.61,128.29,12 6.64,125.56,125.00,123.76,120.62,113.46,106.56,61.06,56.19,47.89.HRMS(ESI)cald.for C 22 H 20 N2O5SNa + [M+Na] + :447.09851,found:447.09852.

[0047] Example 2

[0048] The synthesis of 7-(thiophen-3-yl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-2) in this example is as follows:

[0049]

[0050] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0051] (2) Synthesis of 7-(thiophen-3-yl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-2)

[0052] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 3-thiopheneboronic acid (73.7 mg, 0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-2), yielding 78 mg of a pale white solid in a 38.3% yield. Melting point: 204.5-206.2°C. 1 H NMR (300MHz, CDCl3) δ9.94 (s, 1H), 7.44-7.37 (m, 2H), 7.31 (dd, J = 5.0, 1.2Hz, 1H), 7.23 (d, J = 1.7Hz, 1H) ,7.07(dd,J=8.5,1.7Hz,1H),6.77(d,J=7.6Hz,1H),6.68(s,2H),4.65(s,2H),3.87(s,3H),3.71(s,6H). 13 C NMR (75MHz, CDCl3) δ168.98,168.71,152.99,140.63,140.51,134.05,130.24,128.68,126.79,12 6.41,125.98,124.92,121.10,120.95,114.12,106.62,61.03,56.19,47.90.HRMS(ESI)cald.for C 22 H 20 N2O5SNa + [M+Na] + :447.09851,found:447.09850.

[0053] Example 3

[0054] The synthesis of 7-(furan-2-yl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-3) in this example is as follows:

[0055]

[0056] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0057] (2) Synthesis of 7-(furan-2-yl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-3)

[0058] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), furan-2-boronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-3), yielding 82 mg of a pale white solid in a 41.8% yield. Melting point: 193.6-194.7°C. 1 H NMR (300MHz, DMSO-d6) δ10.87(s,1H),7.77-7.73(m,1H),7.34(d,J=1.5Hz,1H),7.19(d,J=8.7Hz,1H),6. 90(s,1H),6.84(d,J=3.3Hz,1H),6.71(s,2H),6.60-6.56(m,1H),4.41(s,2H),3.69(s,3H),3.66(s,6H). 13 C NMR(75MHz,DMSO-d6)δ168.46,167.08,152.99,152.63,143.54,139.93,132.06,129.90,128.15,1 26.46,125.13,117.59,112.63,111.04,106.71,106.54,60.64,56.39,48.53.HRMS(ESI)cald.for C 22 H 20 N2O6Na + [M+Na] + :431.12136,found:431.12158.

[0059] Example 4

[0060] The synthesis of 7-(furan-3-yl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-4) in this example is as follows:

[0061]

[0062] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0063] (2) Synthesis of 7-(furan-3-yl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-4)

[0064] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), furan-3-boronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-4), yielding 86 mg of a pale white solid in a 43.8% yield. Melting point: 194.3-195.5°C. 1 H NMR(300MHz,DMSO-d6)δ10.81(s,1H),8.08(s,1H),7.76-7.73(m,1H),7.19(d,J=1.3Hz,1H), 7.09(d,J=8.1Hz,1H),6.92-6.79(m,2H),6.71(s,2H),4.40(s,2H),3.69(s,3H),3.66(s,6H). 13 C NMR(75MHz,DMSO-d6)δ168.43,167.12,152.97,144.96,139.86,139.74,132.04,129.97,129.93,1 26.19,125.48,125.05,119.59,113.27,109.02,106.70,60.64,56.38,48.43.HRMS(ESI)cald.for C 22 H 20 N2O6Na + [M+Na] + :431.12136,found:431.12188.

[0065] Example 5

[0066] The synthesis of 7-(2-hydroxyphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-5) in this example is as follows:

[0067]

[0068] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0069] (2) Synthesis of 7-(2-hydroxyphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-5)

[0070] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 2-hydroxyphenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-5), yielding 98 mg of a pale white solid in a 47% yield. Melting point: 149.8-152.1°C. 1 H NMR (300MHz, DMSO-d6) δ10.84(s,1H),9.60(s,1H),7.30(s,1H),7.18(t,J=7.1Hz,2H ),7.04-6.93(m,2H),6.92-6.82(m,2H),6.72(s,2H),4.45(s,2H),3.80-3.54(m,9H). 13 C NMR(75MHz,DMSO-d6)δ168.35,167.29,154.76,152.93,139.95,136.70,131.18,130.41,129.79,129.16,1 27.20,125.93,124.29,122.88,119.88,117.21,116.49,106.91,60.64,56.36,48.57.HRMS(ESI)cald.forC 24 H 22 N2O6Na + [M+Na] + :457.13701found:457.13710.

[0071] Example 6

[0072] The synthesis of 7-(3-hydroxyphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-6) in this example is as follows:

[0073]

[0074] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0075] (2) Synthesis of 7-(3-hydroxyphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-6)

[0076] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 3-hydroxyphenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-6), yielding 103 mg of a pale white solid in a 49.3% yield. Melting point: 145.4-146.9°C. 1 H NMR(300MHz,DMSO-d6)δ10.89(s,1H),9.59(s,1H),7.27-7.21(m,2H),7.08(d,J=8.7Hz,1H), 7.00-6.91(m,3H),6.77(d,J=8.2Hz,1H),6.72(s,2H),4.43(s,2H),3.70(s,3H),3.67(s,6H). 13 C NMR(75MHz,DMSO-d6)δ168.43,167.18,158.30,152.95,141.02,139.89,138.31,132.04,130.54,129.81,1 26.71,125.12,120.36,117.58,115.14,114.36,113.59,106.75,60.65,60.23,56.34.HRMS(ESI)cald.for C 24 H 22 N2O6Na + [M+Na] + :457.13701found:457.13734.

[0077] Example 7

[0078] The synthesis of 7-(4-hydroxyphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-7) in this example is as follows:

[0079]

[0080] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0081] (2) Synthesis of 7-(4-hydroxyphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-7)

[0082] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 4-hydroxyphenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-7), yielding 99 mg of a pale white solid in a 47.4% yield. Melting point: 231.6-233.1°C. 1 H NMR (300MHz, DMSO-d6) δ10.90(s,1H),9.68(s,1H),7.44(d,J=8.6Hz,2H),7.26(d,J=1.6Hz,1H), 7.11(d,J=7.9Hz,1H),6.89(d,J=8.6Hz,2H),6.76(s,2H),4.47(s,2H),3.74(s,3H),3.71(s,6H). 13 CNMR(75MHz,DMSO-d6)δ168.39,167.19,157.78,152.94,139.81,138.29,132.01,130.31,129.92, 127.93,125.81,125.06,119.78,116.27,113.66,106.70,60.63,56.33,48.38.HRMS(ESI)cald.for C 24 H 22 N2O6Na + [M+Na] + :457.13701found:457.13790.

[0083] Example 8

[0084] The synthesis of 7-(2-(hydroxymethyl)phenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-8) in this example is as follows:

[0085]

[0086] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0087] (2) Synthesis of 7-(2-(hydroxymethyl)phenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-8)

[0088] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 2-hydroxymethylphenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-8), yielding 91 mg of a pale white solid in a 42.3% yield. Melting point: 158.8-160.6°C. 1 H NMR(300MHz, CDCl3)δ9.70(s,1H),7.55-7.49(m,1H),7.40-7.29(m,2H),7.21-7.15(m,1H), 7.11(s,1H),6.83(s,2H),6.67(s,2H),4.58(s,2H),4.54(s,2H),3.86(s,3H),3.72(s,6H). 13 C NMR (75MHz, CDCl3) δ168.88,168.65,152.98,140.64,139.83,138.91,137.75,129.84,129.71,129.16,128.6 0,128.25,128.01,126.57,124.42,123.90,117.23,106.64,62.95,61.03,56.19,48.03.HRMS(ESI)cald.for C 25 H 24 N2O6Na + [M+Na]+ :471.15266found:471.15362.

[0089] Example 9

[0090] The synthesis of 7-(3-(hydroxymethyl)phenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-9) in this example is as follows:

[0091]

[0092] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0093] (2) Synthesis of 7-(3-(hydroxymethyl)phenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-9)

[0094] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 3-hydroxymethylphenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-9), yielding 94 mg of a pale white solid in a 43.6% yield. Melting point: 161.2-162.6°C. 1 H NMR (300MHz, DMSO-d6) δ10.85(s,1H),7.55(s,1H),7.46-7.41(m,2H),7.35-7.29(m,2H),7.15(d,J=7.3Hz,1H) ,6.98(s,1H),6.74(s,2H),5.27(t,J=5.6Hz,1H),4.57(d,J=5.5Hz,2H),4.44(s,2H),3.71(s,3H),3.68(s,6H). 13C NMR(75MHz,DMSO-d6)δ168.48,167.15,152.99,143.83,139.99,139.39,138.29,132.12,129.85,129.26, 126.76,126.22,125.15,124.77,120.44,114.42,106.81,63.27,60.65,56.39,48.60.HRMS(ESI)cald.for C 25 H 24 N2O6Na + [M+Na] + :471.15266,found:471.15293.

[0095] Example 10

[0096] The synthesis of 7-(4-(hydroxymethyl)phenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-10) in this example is as follows:

[0097]

[0098] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0099] (2) Synthesis of 7-(4-(hydroxymethyl)phenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-10)

[0100] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 4-hydroxymethylphenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-10), yielding 96 mg of a pale white solid in a 44.6% yield. Melting point: 197.3-198.6°C. 1H NMR (300MHz, DMSO-d6) δ10.84(s,1H),7.53(d,J=8.2Hz,2H),7.41(d,J=8.0Hz,2H),7.31(d,J=1.7Hz,1H),7.15(dd,J=8.6, 1.3Hz,1H),6.96(s,1H),6.72(s,2H),5.22(t,J=5.8Hz,1H),4.54(d,J=5.6Hz,2H),4.43(s,2H),3.71(s,3H),3.67(s,6H). 13 C NMR (75MHz, DMSO-d6) δ168.45,167.14,152.98,142.56,139.98,138.09,137.96,132.12,129.85,129. 52,127.57,126.63,126.51,125.14,114.30,106.83,63.01,60.66,56.39,48.57.HRMS(ESI)cald.forC 25 H 24 N2O6Na + [M+Na] + :471.15266,found:471.15301.

[0101] Example 11

[0102] The synthesis of 7-(2-chlorophenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-11) in this example is as follows:

[0103]

[0104] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0105] (2) Synthesis of 7-(2-chlorophenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-11)

[0106] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 2-chlorophenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-11), yielding 87 mg of a pale white solid in a 40% yield. Melting point: 169.1-170.2°C. 1 H NMR (300MHz, CDCl3) δ9.15 (s, 1H), 7.50-7.43 (m, 1H), 7.33-7.27 (m, 3H), 7.06 (d, J = 1.7Hz, 1H), 6.9 0(dd,J=8.3,1.7Hz,1H),6.81(d,J=8.2Hz,1H),6.68(s,2H),4.65(s,2H),3.86(s,3H),3.72(s,6H). 13 CNMR (126MHz, CDCl3) δ168.63,168.50,152.94,140.68,138.77,137.56,132.32,130.99,130.11,129.42,1 29.15,128.39,127.06,127.04,124.37,124.28,117.26,106.72,61.00,56.14,47.80.HRMS(ESI)cald.for C 24 H 21 N2O5ClNa + [M+Na] + :475.10312,found:475.10439.

[0107] Example 12

[0108] The synthesis of 7-(3-chlorophenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-12) in this example is as follows:

[0109]

[0110] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0111] (2) Synthesis of 7-(2-chlorophenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-11)

[0112] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 3-chlorophenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. Upon completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-12), yielding 92 mg of a pale white solid in a 42.3% yield. Melting point: 169.3-170.5°C. 1 H NMR (300MHz, CDCl3) δ9.08 (s, 1H), 7.51-7.48 (m, 1H), 7.41-7.32 (m, 3H), 7.15 (d, J = 1.7Hz, 1H), 7.0 5(dd,J=8.4,1.7Hz,1H),6.87(d,J=8.2Hz,1H),6.68(s,2H),4.63(s,2H),3.88(s,3H),3.72(s,6H). 13 CNMR(75MHz, CDCl3)δ168.73,168.29,153.00,141.11,140.72,137.92,134.91,130.23,130.18,128.50,1 27.93,127.28,126.94,125.08,124.97,121.77,114.62,106.57,61.01,56.16,48.01.HRMS(ESI)cald.for C 24 H 21 N2O5ClNa + [M+Na] + :475.10312,found:475.10399.

[0113] Example 13

[0114] The synthesis of 7-(4-chlorophenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-13) in this example is as follows:

[0115]

[0116] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0117] (2) Synthesis of 7-(4-chlorophenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-13)

[0118] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 4-chlorophenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. Upon completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-13), yielding 112 mg of a pale white solid in a 51.5% yield. Melting point: 209.7-210.9°C. 1 H NMR (300MHz, DMSO-d6) δ10.88(s,1H),7.59(d,J=8.4Hz,2H),7.52(d,J=8.4Hz,2H),7.28(s,1H) ,7.16(d,J=8.2Hz,1H),7.03-6.91(m,1H),6.73(s,2H),4.42(s,2H),3.70(s,3H),3.67(s,6H). 13 C NMR(75MHz,DMSO-d6)δ168.51,167.10,152.99,139.98,138.41,136.72,132.97,132.18,129.81,1 29.49,128.56,127.09,125.23,120.44,114.38,106.78,60.65,56.39,48.59.HRMS(ESI)cald.for C 24 H 21 N2O5ClNa + [M+Na] + :475.10312,found:475.10393.

[0119] Example 14

[0120] The synthesis of 7-(2-methylphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-14) in this example is as follows:

[0121]

[0122] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0123] (2) Synthesis of 7-(2-methylphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-14)

[0124] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 2-methylphenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-14), yielding 101 mg of a pale white solid in a 48.6% yield. Melting point: 190.4-192.6°C. 1 H NMR(300MHz,DMSO-d6)δ10.86(s,1H),7.31-7.23(m,3H),7.17-7.12(m,1H),7.01-6.97(m ,1H),6.90-6.77(m,2H),6.72(s,2H),4.46(s,2H),3.70(s,3H),3.68(s,6H),2.20(s,3H). 13 C NMR (75MHz, DMSO-d6) δ168.42,167.29,152.94,140.73,139.86,139.21,135.05,131.46,130.90,129.80,129. 66,128.01,126.52,126.26,124.46,122.81,116.85,106.75,60.63,56.29,48.32,20.44.HRMS(ESI)cald.for C 25 H 24 N2O5Na + [M+Na] + :455.15774,found:455.15788.

[0125] Example 15

[0126] The synthesis of 7-(3-methylphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-15) in this example is as follows:

[0127]

[0128] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0129] (2) Synthesis of 7-(3-methylphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-15)

[0130] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 3-methylphenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-15), yielding 93 mg of a pale white solid in a 44.8% yield. Melting point: 169.9-171.2°C. 1 H NMR(300MHz, CDCl3)δ8.91(s,1H),7.34-7.29(m,3H),7.21-7.15(m,2H),7.06(dd,J=8.5,1.8H z,1H),6.82(d,J=8.8Hz,1H),6.67(s,2H),4.63(s,2H),3.87(s,3H),3.71(s,6H),2.41(s,3H). 13 C NMR (75MHz, CDCl3) δ168.69,168.32,152.99,140.70,139.64,139.32,138.71,130.07,128.92,128.72,128.6 4,127.59,126.71,124.96,123.95,121.84,114.61,106.70,61.02,56.17,47.99,21.55.HRMS(ESI)cald.for C 25 H 24 N2O5Na + [M+Na] + :455.15774,found:455.15882.

[0131] Example 16

[0132] The synthesis of 7-(4-methylphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-16) in this example is as follows:

[0133]

[0134] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0135] (2) Synthesis of 7-(4-methylphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-16)

[0136] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 4-methylphenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-16), yielding 98 mg of a pale white solid in a 47.2% yield. Melting point: 246.5-248.2°C. 1 H NMR(300MHz, CDCl3)δ8.36(s,1H),7.41(d,J=7.9Hz,2H),7.26-7.22(m,2H),7.13-7.10(m,1H),7.06(dd,J =8.2,1.6Hz,1H),6.83(d,J=7.5Hz,1H),6.67(s,2H),4.62(s,2H),3.87(s,3H),3.71(s,6H),2.39(s,3H). 13 C NMR (75MHz, CDCl3) δ168.67,168.35,152.98,140.71,139.46,137.92,136.44,130.11,129.72,128.65,126.6 6,126.52,126.18,124.98,124.77,121.59,114.37,106.72,61.01,56.17,48.00,21.13.HRMS(ESI)cald.for C 25 H24 N2O5Na + [M+Na] + :455.15774,found:455.15835.

[0137] Example 17

[0138] The synthesis of 2-(3-oxo-1-(3,4,5-trimethoxybenzoyl)-1,2,3,4-tetrahydroquinoxaline-6-yl)benzyl cyanide (I-17) in this example is as follows:

[0139]

[0140] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0141] (2) Synthesis of 2-(3-oxo-1-(3,4,5-trimethoxybenzoyl)-1,2,3,4-tetrahydroquinoxalin-6-yl)benzyl cyanide (Ⅰ-17)

[0142] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 2-cyanophenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-17), yielding 89 mg of a pale white solid in a 41.8% yield. Melting point: 230-232.2°C. 1 H NMR (300MHz, DMSO-d6) δ11.01 (s, 1H), 7.96 (d, J = 7.6Hz, 1H), 7.84-7.76 (m, 1H), 7.64-7.53 ( m,2H),7.21(s,1H),7.07-6.94(m,2H),6.73(s,2H),4.47(s,2H),3.69(s,3H),3.68(s,6H). 13C NMR (75MHz, DMSO-d6) δ168.51,167.24,152.96,144.06,139.91,135.63,134.38,134.12,131.78,130.32,129. 58,128.91,127.83,125.00,122.58,118.72,116.56,110.54,106.76,60.62,56.34,48.30.HRMS(ESI)cald.for C 25 H 21 N3O5Na + [M+Na] + :466.13734,found:466.13773.

[0143] Example 18

[0144] The synthesis of 3-(3-oxo-1-(3,4,5-trimethoxybenzoyl)-1,2,3,4-tetrahydroquinoxaline-6-yl)benzyl cyanide (I-18) in this example is as follows:

[0145]

[0146] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0147] (2) Synthesis of 3-(3-oxo-1-(3,4,5-trimethoxybenzoyl)-1,2,3,4-tetrahydroquinoxalin-6-yl)benzyl cyanide (Ⅰ-18)

[0148] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 3-cyanophenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. Upon completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-18), yielding 91 mg of a pale white solid in a 42.7% yield. Melting point: 191.1-192.2°C. 1H NMR (300MHz, CDCl3) δ9.27(s,1H),7.82-7.78(m,1H),7.77-7.72(m,1H),7.66(d,J=7.6Hz,1H),7.56(d,J=7.7Hz,1H),7.1 6(d,J=1.6Hz,1H),7.06(dd,J=8.4,1.1Hz,1H),6.93(d,J=8.5Hz,1H),6.69(s,2H),4.64(s,2H),3.88(s,3H),3.73(s,6H). 13 C NMR (75MHz, CDCl3) δ168.84,168.30,153.11,140.93,140.65,136.89,131.28,131.15,130.46,130.37,129.90 ,128.46,127.82,125.29,121.76,118.54,114.69,113.30,106.64,61.04,56.24,48.08.HRMS(ESI)cald.forC 25 H 21 N3O5Na + [M+Na] + :466.13734,found:466.13796.

[0149] Example 19

[0150] The synthesis of 4-(3-oxo-1-(3,4,5-trimethoxybenzoyl)-1,2,3,4-tetrahydroquinoxaline-6-yl)benzyl cyanide (I-19) in this example is as follows:

[0151]

[0152] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0153] (2) Synthesis of 4-(3-oxo-1-(3,4,5-trimethoxybenzoyl)-1,2,3,4-tetrahydroquinoxalin-6-yl)benzyl cyanide (Ⅰ-19)

[0154] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 4-cyanophenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-19), yielding 85 mg of a pale white solid in a 39.9% yield. Melting point: 205.7-206.9°C. 1 H NMR (300MHz, DMSO-d6) δ10.93(s,1H),7.93(d,J=8.3Hz,2H),7.78(d,J=8.3Hz,2H),7.36(d,J=1.8Hz,1 H),7.26(dd,J=8.4,1.6Hz,1H),7.07-6.98(m,1H),6.74(s,2H),4.43(s,2H),3.70(s,3H),3.67(s,6H). 13 C NMR(75MHz,DMSO-d6)δ167.05,153.01,144.02,140.03,135.99,133.47,132.27,129.75,127.9 2,127.65,125.32,123.65,120.90,114.79,110.62,106.78,60.66,56.41.HRMS(ESI)cald.for C 25 H 21 N3O5Na + [M+Na] + :466.13734,found:466.13897.

[0155] Example 20

[0156] The synthesis of 7-(2-fluorophenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-20) in this example is as follows:

[0157]

[0158] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0159] (2) Synthesis of 7-(2-fluorophenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-20)

[0160] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 2-fluorophenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-20), yielding 79 mg of a pale white solid in a 37.7% yield. Melting point: 217.9-218.2°C. 1 H NMR(300MHz,DMSO-d6)δ10.91(s,1H),7.51-7.41(m,2H),7.36-7.27(m,2H),7.2 3(s,1H),7.06-6.93(m,2H),6.72(s,2H),4.45(s,2H),3.70(s,3H),3.68(s,6H). 13 C NMR(75MHz,DMSO-d6)δ168.47,167.21,159.44(d, 1 J C-F =244.30Hz),152.96,139.98,132.89,131.78,130.89(d, 3 J C-F =3.14Hz),130.25(d, 3 J C-F =8.44Hz),129.68,127.77(d, 2 J C-F =13.07Hz),127.07,125.51(d, 4 J C-F =3.47Hz),124.92,122.53,116.74,116.65(d, 2 J C-F =22.00Hz),106.85,60.65,56.34,48.45.HRMS(ESI)cald.for C 24 H 21 N2O5FNa + [M+Na] +:459.13267,found:459.13327.

[0161] Example 21

[0162] The synthesis of 7-(3-fluorophenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-21) in this example is as follows:

[0163]

[0164] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0165] (2) Synthesis of 7-(3-fluorophenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-21)

[0166] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 3-fluorophenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-21), yielding 76 mg of a pale white solid in a 36.2% yield. Melting point: 180.6-182.3°C. 1 H NMR(300MHz, CDCl3)δ9.41(s,1H),7.45-7.35(m,1H),7.32-7.26(m,1H),7.24-7.16(m,2H),7 .10-7.01(m,2H),6.86(d,J=7.4Hz,1H),6.68(s,2H),4.64(s,2H),3.87(s,3H),3.72(s,6H). 13 C NMR(75MHz,CDCl3)δ168.77,168.54,163.21(d, 1 J C-F =245.18Hz),153.04,141.58(d, 3 J C-F =7.71Hz),140.80,138.08,130.55(d, 3 J C-F=8.49Hz),130.27,128.54,127.29,125.06,122.49(d, 4 J C-F =2.71Hz),121.76,114.77(d, 2 J C-F =21.04Hz),114.73,113.79(d, 2 J C-F =21.98Hz),106.68,61.03,56.20,48.00.HRMS(ESI)cald.for C 24 H 21 N2O5FNa + [M+Na] + :459.13267,found:459.13326.

[0167] Example 22

[0168] The synthesis of 7-(4-fluorophenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-22) in this example is as follows:

[0169]

[0170] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0171] (2) Synthesis of 7-(4-fluorophenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-22)

[0172] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 4-fluorophenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-22), yielding 71 mg of a pale white solid in a 33.9% yield. Melting point: 204.4-205.9°C. 1H NMR (300MHz, CDCl3) δ9.86 (s, 1H), 7.54-7.42 (m, 2H), 7.20-7.07 (m, 3H), 7.02 (d, J = 8 .1Hz,1H),6.84(d,J=7.1Hz,1H),6.68(s,2H),4.65(s,2H),3.87(s,3H),3.71(s,6H). 13 C NMR (75MHz, CDCl3) δ168.93,168.75,162.76(d, 1 J C-F =246.23Hz),153.02,140.76,138.47,135.49(d, 4 J C-F =2.99Hz),130.31,128.60,128.47(d, 3 J C-F =8.09Hz),126.75,124.98,121.64,115.93(d, 2 J C-F =21.50Hz),114.69,106.71,61.02,56.19,47.96.HRMS(ESI)cald.for C 24 H 21 N2O5FNa + [M+Na] + :459.13267,found:459.13319.

[0173] Example 23

[0174] The synthesis of 7-(2-methoxyphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-23) in this example is as follows:

[0175]

[0176] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0177] (2) Synthesis of 7-(2-methoxyphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-23)

[0178] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 2-methoxyphenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-23), yielding 81 mg of a pale white solid in a 37.6% yield. Melting point: 181.3-183.1°C. 1 H NMR(300MHz, CDCl3)δ8.96(s,1H),7.39-7.30(m,1H),7.24-7.20(m,1H),7.15(s,1H),7.05-6.93 (m,3H),6.77(d,J=7.5Hz,1H),6.67(s,2H),4.63(s,2H),3.86(s,3H),3.80(s,3H),3.71(s,6H). 13 C NMR (75MHz, CDCl3) δ168.60,168.50,156.32,152.89,140.60,136.94,130.45,129.38,129.30,128.85,128.6 0,126.33,124.34,124.31,120.98,117.27,111.27,106.82,61.02,56.11,55.52,47.88.HRMS(ESI)cald.for C 24 H 24 N2O6Na + [M+Na] + :471.15266,found:471.15328.

[0179] Example 24

[0180] The synthesis of 7-(3-methoxyphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-24) in this example is as follows:

[0181]

[0182] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0183] (2) Synthesis of 7-(3-methoxyphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-24)

[0184] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 3-methoxyphenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-24), yielding 79 mg of a pale white solid in a 36.7% yield. Melting point: 187.8-189.3°C. 1 H NMR(300MHz, CDCl3)δ9.12(s,1H),7.40-7.30(m,1H),7.18(s,1H),7.12-7.01(m,3H),6.91( d,J=7.4Hz,1H),6.83(d,J=7.8Hz,1H),6.68(s,2H),4.63(s,2H),3.86(s,6H),3.71(s,6H). 13 C NMR (75MHz, CDCl3) δ168.71,168.38,160.08,153.01,140.85,140.72,139.32,130.13,130.05,128.63,126.9 1,124.95,121.83,119.32,114.73,113.09,112.83,106.70,61.02,56.18,55.41,48.00.HRMS(ESI)cald.for C 24 H 24 N2O6Na + [M+Na] + :471.15266,found:471.15332.

[0185] Example 25

[0186] The synthesis of 7-(4-methoxyphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-25) in this example is as follows:

[0187]

[0188] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0189] (2) Synthesis of 7-(4-methoxyphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-25)

[0190] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 4-methoxyphenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-25), yielding 85 mg of a pale white solid in a 39.4% yield. Melting point: 230.1-232.3°C. 1 H NMR (300MHz, CDCl3) δ9.82 (s, 1H), 7.45 (d, J = 7.8Hz, 2H), 7.18 (s, 1H), 7.06-6.91 (m, 3H ),6.86-6.74(m,1H),6.68(s,2H),4.64(s,2H),3.87(s,3H),3.84(s,3H),3.70(s,6H). 13 C NMR (75MHz, CDCl3) δ168.97,168.69,159.63,152.97,140.66,139.13,131.81,130.25,128.70,127.9 0,126.15,124.90,121.28,114.43,114.29,106.73,61.01,56.18,55.42,47.95.HRMS(ESI)cald.forC 24 H 24 N2O6Na + [M+Na] + :471.15266,found:471.15306.

[0191] Example 26

[0192] The synthesis of 7-phenyl-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-26) in this example is as follows:

[0193]

[0194] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0195] (2) Synthesis of 7-phenyl-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-26)

[0196] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), phenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. Upon completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-26), yielding 92 mg of a pale white solid in a 45.8% yield. Melting point: 211.6-213.2°C. 1 H NMR (300MHz, CDCl3) δ9.43 (s, 1H), 7.54-7.49 (m, 2H), 7.48-7.40 (m, 2H), 7.40-7.36 (m, 1H), 7.20 (d, J = 1.7Hz ,1H),7.08(dd,J=8.4,1.6Hz,1H),6.83(d,J=8.3Hz,1H),6.68(s,2H),4.65(s,2H),3.87(s,3H),3.71(s,6H). 13 C NMR (75MHz, CDCl3) δ168.72,168.69,152.97,140.62,139.49,139.33,130.17,129.03,128.61,12 7.97,126.86,126.75,125.00,121.82,114.74,106.63,61.05,56.16,47.95.HRMS(ESI)cald.for C 24 H 22 N2O5Na + [M+Na] + :441.14209,found:441.14336.

[0197] Example 27

[0198] The synthesis of 7-(naphthalen-1-yl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-27) in this example is as follows:

[0199]

[0200] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0201] (2) Synthesis of 7-(naphthalen-1-yl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (I-27)

[0202] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 1-naphthaleneboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. Upon completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-27), yielding 105 mg of a pale white solid in a 45.4% yield. Melting point: 133.5-134.2°C. 1 H NMR (300MHz, DMSO-d6) δ10.87(s,1H),8.02(d,J=7.4Hz,1H),7.97(d,J=8.2Hz,1H),7.75(d,J=8.2Hz,1H),7.62- 7.48(m,3H),7.40(d,J=6.9Hz,1H),7.16-7.12(m,1H),7.02-6.90(m,2H),6.76(s,2H),4.50(s,2H),3.72(s,9H). 13 C NMR(75MHz,DMSO-d6)δ168.50,167.31,153.02,140.09,138.95,137.94,133.89,131.74,131.21,129.81,128.93,128.34,1 27.08,126.84,126.77,126.52,126.04,125.34,124.78,123.53,117.61,106.94,60.68,56.42,48.44.HRMS(ESI)cald.forC 28 H 24 N2O5Na +[M+Na] + :491.15774,found:491.15947.

[0203] Example 28

[0204] The synthesis of 7-(naphthalen-2-yl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-28) in this example is as follows:

[0205]

[0206] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0207] (2) Synthesis of 7-(naphthalen-2-yl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (I-28)

[0208] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 2-naphthaleneboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-28), yielding 112 mg of a pale white solid in a 49.8% yield. Melting point: 166.9-168.2°C. 1 H NMR (300MHz, CDCl3) δ9.37(s,1H),7.98(s,1H),7.94-7.84(m,3H),7.65(dd,J=8.4,1.1Hz,1H),7.55-7.47(m,2H),7.33( d,J=1.4Hz,1H),7.21(dd,J=8.4,1.1Hz,1H),6.88(d,J=8.0Hz,1H),6.70(s,2H),4.67(s,2H),3.88(s,3H),3.72(s,6H). 13C NMR (75MHz, CDCl3) δ168.75,168.60,153.01,140.65,139.37,136.58,133.55,132.83,130.25,128.80,128.65,128.24,12 7.72,126.84,126.67,126.42,125.72,125.09,124.89,122.08,114.93,106.65,61.06,56.19,48.00.HRMS(ESI)cald.for C 28 H 24 N2O5Na + [M+Na] + :491.15774,found:491.15909.

[0209] Example 29

[0210] The synthesis of 7-(3,5-dichlorophenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-29) in this example is as follows:

[0211]

[0212] (1) The preparation methods of compound III-1, compound IV-1 and compound V-1 are the same as those in Example 1.

[0213] (2) Synthesis of 7-(naphthalen-2-yl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (I-28)

[0214] Under nitrogen, compound V-1 (200 mg, 0.48 mmol), 3,5-dichlorophenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-29), yielding 89 mg of a pale white solid in a 38% yield. Melting point: 228.3-230.1°C. 1H NMR (300MHz, CDCl3) δ9.69 (s, 1H), 7.41-7.33 (m, 3H), 7.15 (s, 1H), 7.02 (d, J = 8.0H z,1H),6.89(d,J=8.0Hz,1H),6.68(s,2H),4.64(s,2H),3.88(s,3H),3.73(s,6H). 13 C NMR (75MHz, CDCl3) δ168.84,168.64,153.09,142.31,140.89,136.55,135.57,130.41,128.46,12 7.82,127.78,125.35,125.14,121.76,114.77,106.63,61.04,56.23,48.02.HRMS(ESI)cald.for C 24 H 20 N2O5Cl2Na + [M+Na] + :509.06415,found:509.06572.

[0215] Example 30

[0216] The synthesis of 6-phenyl-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-30) in this example is as follows:

[0217]

[0218] (1) Synthesis of (5-bromo-2-nitrophenyl)glycine methyl ester (Ⅲ-2)

[0219] The difference from Example 1 is that II-2 is 2-fluoro-4-bromonitrobenzene. The rest of the operation is the same as that of Intermediate III-1, and 6.1 g of light yellow solid is obtained with a yield of 93%. Melting point: 103.3-104.4°C. 1 H NMR (300MHz, CDCl3) δ8.44(br,1H),8.06(d,J=9.5Hz,1H),6.90-6.81(m,3H),4.09(d,J=5.2Hz,2H),3.85(s,3H).

[0220] (2) Synthesis of 6-bromo-3,4-dihydroquinoxaline-2(1H)-one (Ⅳ-2)

[0221] The same operation as that of Intermediate IV-1 was used to obtain 1.2 g of a brown solid with a yield of 76.4%. Melting point: 172.3-173.6°C. 1H NMR (300MHz, DMSO-d6) δ10.36(br,1H),6.82-6.79(m,1H),6.74-6.69(m,1H),6.67-6.61(m,1H),6.22(br,1H),3.76(s,2H).

[0222] (3) Synthesis of 6-bromo-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (V-2)

[0223] The same operation as that of Intermediate V-1 was used to obtain 833 mg of a brown-yellow solid with a yield of 44.9%. Melting point: 182.0-183.2°C. 1 H NMR (300MHz, CDCl3) δ9.15 (s, 1H), 7.29-7.25 (m, 1H), 7.00 (s, 1H), 6.86 (d, J = 8.3Hz, 1H), 6.65 (s, 2H), 4.55 (s, 2H), 3.88 (s, 3H), 3.75 (s, 6H). 13 C NMR (75MHz, CDCl3) δ168.71,167.93,153.15,141.22,129.03,128.93,128.75,12 7.99,127.59,117.53,115.27,106.70,61.08,56.35,47.96.HRMS(ESI)cald.for C 18 H 17 N2O5BrNa + [M+Na] + :443.02131,found:443.02178.

[0224] (4) Synthesis of 6-phenyl-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-30)

[0225] Phenylboronic acid was selected as aromatic boronic acid VI, and the operation process was the same as that of target compound I-1 to obtain 72 mg of light white solid with a yield of 35.8%. Melting point: 107.8-109.2℃. 1 H NMR(300MHz, CDCl3)δ8.87(s,1H),7.36-7.28(m,3H),7.15-7.07(m,2H),7.03(d ,J=8.8Hz,1H),6.95(s,1H),6.70(s,2H),4.66(s,2H),3.86(s,3H),3.71(s,6H). 13C NMR (75MHz, CDCl3) δ168.65,168.11,153.12,140.77,139.62,136.48,128.86,128.84,127.7 9,127.59,126.69,124.87,123.63,116.53,106.83,60.99,56.31,47.68.HRMS(ESI)cald.for C 24 H 22 N2O5Na + [M+Na] + :441.14209,found:441.14261.

[0226] Example 31

[0227] The synthesis of 6-(3-hydroxyphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxaline-2(1H)-one (I-31) in this example is as follows:

[0228]

[0229] (1) The preparation methods of compound III-2, compound IV-2 and compound V-2 are the same as those in Example 30.

[0230] (2) Synthesis of 6-(3-hydroxyphenyl)-4-(3,4,5-trimethoxybenzoyl)-3,4-dihydroquinoxalin-2(1H)-one (Ⅰ-31)

[0231] Under nitrogen, compound V-2 (200 mg, 0.48 mmol), 3-hydroxyphenylboronic acid (0.576 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (35.1 mg, 0.048 mmol), and potassium carbonate (132.7 mg, 0.96 mmol) were dissolved in 1,4-dioxane (10 mL). 0.1 mL of water was added and the mixture was allowed to fully dissolve. The reaction mixture was then allowed to react at 90°C for 24 h. The reaction was monitored by TLC. After completion, the mixture was cooled to room temperature and evaporated under reduced pressure. The mixture was then purified by column chromatography (DCM:MeOH = 50:1) to afford compound (I-31), yielding 94 mg of a white solid in a 45% yield. Melting point: 131.3-133.2°C. 1H NMR (300MHz, CDCl3) δ9.29 (s, 1H), 7.27-7.25 (m, 2H), 7.20-7.14 (m, 1H), 7.00 (d, J = 8.2Hz, 1H), 6.80- 6.74(m,2H),6.71(d,J=7.9Hz,1H),6.68(s,2H),6.33(s,1H),4.67(s,2H),3.91(s,3H),3.69(s,6H). 13 C NMR (75MHz, CDCl3) δ168.79,168.30,156.32,153.45,141.22,140.08,136.24,130.08,129.39,128.91,12 7.42,124.89,123.47,118.60,116.54,114.67,114.02,106.65,61.15,56.35,47.36.HRMS(ESI)cald.for C 24 H 22 N2O6Na + [M+Na] + :457.13701,found:457.13720.

[0232] Application Examples

[0233] Below are some pharmacological experiments and results described in the present invention:

[0234] Application Example 1

[0235] The compounds described herein were tested for their inhibitory activity against human cervical cancer cells (HeLa) and human hepatocellular carcinoma cells (HepG2). All cell lines were cryopreserved and passaged in our laboratory. Tumor cells in the logarithmic growth phase were seeded at 4,000 to 5,000 cells / well in a 96-well plate. After 24 hours, various concentrations of the target compound or control drug (compound ABT-751 was selected as the positive control) were added and incubated at 37°C, 5% CO₂ for 72 hours. 50 μL of MTT solution (5 mg / mL) was added to each well. After a further 4 hours of incubation, the culture medium and MTT solution were discarded. 100 μL of DMSO solution was added to each well. After shaking, the absorbance (OD) was measured at 570 nm using a microplate reader. The cell inhibition rate was calculated as follows: Growth inhibition rate = (OD control - OD experimental) / (OD control - OD blank) × 100% (OD control, OD experimental, and OD blank represent the average absorbance values ​​of each group, respectively). The IC₅₀ values ​​of the compounds were then fitted using GraphPad software.

[0236] Experimental results

[0237] The inhibitory activity of the target compounds on tumor cell proliferation is shown in Table 1. Some compounds showed nanomolar-level inhibitory effects. Among them, compounds I-2, I-4, and I-5 showed strong inhibitory activity against both tumor cells, which was slightly weaker than the positive drugs Colchicine and CA-4, but better than DOX.

[0238] Table 1. Anti-cell proliferation activity of compounds

[0239]

[0240] Application Example 2: Microtubule Polymerization Inhibition Experiment

[0241] The inhibitory effects of the compounds on tubulin polymerization were investigated using a tubulin polymerization assay kit (BL011P, Cytoskeleton, USA). Compound I-2 at varying concentrations and the positive controls colchicine and paclitaxel were added to a 96-well plate and incubated at 37°C for 1 min. Tubulin reagent (2.0 mg / mL porcine brain tubulin, 80 mM PIPES pH 6.9, 2.0 mM MgCl2, 0.5 mM EGTA, 1.0 mM GTP, and 15% glycerol) was then added to each well and mixed thoroughly. The compounds were detected every 60 s for 80 min using a multi-function microplate reader (excitation: 360 nm; emission: 450 nm).

[0242] Experimental results

[0243] The inhibitory effect of compound Ⅰ-2 on microtubule polymerization is as follows Figure 1 As shown in the figure, it can be seen that compound I-2 inhibits the polymerization of tubulin like the positive drug Colchicine, while the positive drug Paclitaxel can promote the polymerization of tubulin, indicating that compound I-2 is a tubulin polymerization inhibitor.

[0244] Application Example 3: Cell Cycle Experiment

[0245] HeLa cells were evenly seeded in 6-well plates at appropriate concentrations and incubated at 37°C and 5% CO2 for 24 h. Different concentrations of the test compound were added to each well and incubated for another 24 h. The cells were collected and centrifuged. The supernatant was discarded, 500 μL of pre-cooled PBS solution was added, pipetted evenly, and slowly dripped vertically into 10 mL of pre-cooled 70% (v / v) ethanol. Place at -20°C for fixation for 48 h. Centrifuge, discard the supernatant, add 1 mL of pre-cooled PBS solution, centrifuge, and discard the supernatant. 500 μL of stain (50 μg / mL of PI and 100 μg / mL of RNaseA) was added to each sample.

[0246] Incubate at 4°C in the dark for 1 hour, centrifuge, and discard the dye. Add 1 mL of pre-chilled PBS, centrifuge, and discard the supernatant. Resuspend in 500 μL of PBS, filter through a 300-mesh sieve, and analyze using a flow cytometer at 580 nm.

[0247] Experimental results

[0248] The cell cycle arrest effect of compound Ⅰ-2 is as follows Figure 2 As shown, it can be seen that compound I-2 can arrest the cell cycle at the G2 / M phase at 20 nM, with the amount reaching 66.74%, and its inhibitory effect shows a certain concentration dependence.

[0249] Application Example 4: Docking diagram of Ⅰ-2 and microtubules

[0250] Molecular docking was performed using Maestro 11.5 software to investigate the binding mode of the compounds with microtubules. First, the cocrystal complex of colchicine and microtubules (PDB: 1SA0) was downloaded from the PDB database. Protein preparation options were used to hydrogenate the protein, remove water, add missing residues or fragments, and perform energy minimization. A mesh was then generated using the receptor mesh generation option, following standard procedures recommended by the software. The compounds were hydrogenated, 3D structures were generated, ionized, and optimized using the OPLS3 force field. Molecular docking was then performed using the standard exact mode in Glide Docking.

[0251] Experimental results

[0252] The docking diagram of compound I-2 and microtubules is shown in Figure 3A. In the figure, the 3,4,5-trimethoxyphenyl group occupies the hydrophobic pocket on β-tubulin, one of the oxygen atoms forms a hydrogen bond with βCys241, the tetrahydroquinoxalinone occupies the hydrophobic pocket at the junction of α and β-tubulin, and the thiophene ring extends to the solvent accessible area at the junction of α and β-tubulin. The superposition diagram of compound I-2 and colchicine is shown in Figure 3A. Figure 3 As shown in B, it can be seen that Ⅰ-2 overlaps well with colchicine, and the thiophene ring extends additionally to the junction of α and β tubulin.

[0253] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A biaryl hydrocarbon compound, characterized in that: The general structural formula is shown in formula (I): Among them, the aromatic ring is Any one of the following; where R 1 is any one of H, F, Cl, Br, CH3, CH2CH3, OH, OCH3, CF3, OCF3, CN, CH2OH, NH2, NHCH3 and N(CH3)2, R 1 It is any one of mono-, di-, and tri-substituted.

2. The biaryl hydrocarbon compound according to claim 1, characterized in that The biaromatic hydrocarbon compound is a compound having the following structure:

3. The method for preparing the biaryl hydrocarbon compound according to claim 2, characterized in that: Here are the steps: (1) Compound II undergoes a nucleophilic reaction to generate compound III; (2) Compound III undergoes a reduction reaction with SnCl2 and further undergoes a cyclization reaction to generate compound IV; (3) Compound IV undergoes acylation to generate compound V; (4) Compound V reacts with aromatic boronic acid VI under alkaline conditions and a palladium catalyst to undergo a Suzuki reaction to produce the target product I; Among them, the structural formula of compound II is The structural formula of compound III is The structural formula of compound IV is The structural formula of compound V is 4. The method for preparing the compound according to claim 3, wherein The specific steps for generating compound III in step (1) are: performing a nucleophilic reaction between compound II, glycine methyl ester hydrochloride and a base in a solvent to generate compound III; the specific preparation method of compound V in step (3) is: performing an acylation reaction between compound IV and 3,4,5-trimethoxybenzoyl chloride under the catalysis of DMAP and using anhydrous tetrahydrofuran as solvent to generate compound V.

5. The method for preparing the compound according to claim 4, characterized in that: In the step (4), the aromatic boronic acid VI is 2-thiopheneboronic acid, 3-thiopheneboronic acid, furan-2-boronic acid, furan-3-boronic acid, 2-hydroxyphenylboronic acid, 3-hydroxyphenylboronic acid, 4-hydroxyphenylboronic acid, 2-hydroxymethylphenylboronic acid, 3-hydroxymethylphenylboronic acid, 4-hydroxymethylphenylboronic acid, 2-chlorophenylboronic acid, 3-chlorophenylboronic acid, 4-chlorophenylboronic acid, 2-methylphenylboronic acid, 3-methylphenylboronic acid, 4-methylphenylboronic acid, 2-cyanophenylboronic acid, The palladium catalyst is any one of PdCl2(dppf), Pd(OAc)2 and Pd(PPh3)4.

6. A pharmaceutical composition, characterized in that: Comprising the biaryl hydrocarbon compound according to claim 1 or 2 and a pharmaceutically acceptable carrier.

7. Use of the pharmaceutical composition according to claim 6 in the preparation of a microtubule polymerization inhibitor.

8. The use according to claim 7, characterized in that The microtubule polymerization inhibitor is a drug for treating malignant tumors.

9. The use according to claim 8, characterized in that The malignant tumor is cervical cancer, liver cancer, lung cancer or breast cancer.

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