An inhibitor targeting a homodimer, and its preparation method and application

Abstract

The present invention discloses a quinazolinone-fused triazole ring derivative with a structural general formula (I), and its preparation method is as follows: Using 4-chloro-2-fluorobenzamide as the initial raw material, through six-step reactions, the class I derivative of formula (1) is obtained; by changing R1 of the YM-1 derivative, the synthesized novel compound has an obvious inhibitory effect on the hepatocellular carcinoma cell line HepG2. In addition, this class of compounds targets the MAT2A homodimer and has a significant inhibitory effect on the MAT2A enzyme, where the IC 50 minimum is 0.34 μM.

Description

Technical Field

[0001] The present invention relates to a drug and its preparation method and application, and particularly to a quinazolinone-fused triazole ring derivative targeting homodimer and its preparation method and application. Background Art

[0002] Methionine adenosyltransferase (MAT) is a key enzyme that catalyzes the synthesis of S-adenosylmethionine (SAM) from ATP and Met, and plays an important role in the life process of mammals. MAT2A is an important enzyme for synthesizing non-hepatic endogenous SAM. SAM is the main methyl donor in the methylation processes of nucleic acids, polyamines, and histones. Methylation modification can regulate the activity and expression levels of DNA, RNA, and proteins, and thus control cell growth, differentiation, and death. Studies have shown that MAT2A forms functional homodimers in its purified active form and is widely present in the cytoplasm and nucleus. Compounds co-crystallized with MAT2A protein in the S-adenosylmethionine (MAT2A·SAM) complex, and it was observed that two compounds bind to the interface of the exclusive MAT2A dimer, one molecule for each MAT2A monomer. MAT2A is highly expressed in tumors such as liver cancer, gastric cancer, colon cancer, glioblastoma, and leukemia, and is related to the growth and proliferation of cancer cells. MAT2A participates in various cell metabolic pathways and protein translation regulation through SAM, promotes the growth and proliferation of cancer cells, and accelerates the development of cancer. In addition, studies have shown that downregulating the expression of MAT2A can effectively inhibit the growth of cancer cells during the occurrence and development of hepatocellular carcinoma.

[0003] There are various ways to treat tumors. Surgical treatment, radiotherapy, and chemotherapy are the most commonly used treatment means for human tumor treatment at present, and chemotherapy is still the first-line treatment method for many tumor indications. During the use of traditional chemotherapy drugs, small molecule anti-tumor drugs usually diffuse throughout the body by oral administration or intravenous administration to kill tumor cells in the diseased area. However, during the process of killing tumor cells, it is inevitable to cause damage to normal cells, resulting in obvious toxic and side effects. Due to the obvious drawbacks of traditional chemotherapy methods, new tumor treatment methods such as targeted therapy and gene therapy have developed rapidly in recent years. At present, there is no MAT2A inhibitor on the market, and only some compounds have entered the clinical trial stage. Therefore, it is extremely important to develop MAT2A inhibitors with high activity, low toxicity, multiple structural types, and good physicochemical properties for anti-cancer clinical research.

[0004] According to the drug design strategies of molecular hybridization and bioisosteres, the present invention constructs a novel backbone of quinazolinone-fused triazole ring structure. Through the in vitro and in vivo MAT2A enzyme inhibitory activity tests, compound I7 has good enzyme IC 50= 0.34 ± 0.04 μM, and the inhibition rate of compound I2 against the HepG2 cell line was 54%. This study provides a new design idea for the synthesis of novel backbone MAT2A homodimer inhibitors. SUMMARY OF THE INVENTION

[0005] OBJECT OF THE INVENTION: The first object of the present invention is to provide a quinazolinone-fused triazole ring derivative having an obvious inhibitory effect on MAT2A homodimer; the second object is to provide a preparation method of the quinazolinone-fused triazole ring derivative; the third object is to provide an application of the quinazolinone-fused triazole ring derivative.

[0006] TECHNICAL SOLUTION: The quinazolinone-fused triazole ring derivative of the present invention has the chemical structural formula:

[0007]

[0008] wherein R1 is n is 0 to 6, X is H or a 5- to 7-membered aliphatic heterocycle, the ring heteroatoms are N, O, Y is N, S or a halogen, and L is H, a halogen or R2 is a halogen, an alkoxy group or a hydroxyl group.

[0009] Preferably, the is H,

[0010] Preferably, the is -Cl, -NH2, -SH,

[0011] Preferably, the quinazolinone-fused triazole ring derivative is selected from any of the following compounds:

[0012]

[0013] The preparation method of the LYW-1 derivative of the present invention includes the following steps:

[0014] (1)(2) 4-Chloro-2-fluorobenzamide reacts with oxalyl chloride, and then the benzamide is acylated, and then aniline is added for condensation reaction to obtain compound II;

[0015] (3) Under low-temperature conditions, using the organic base KHMDS as a catalyst, compound II undergoes an intramolecular cyclization reaction to obtain compound III;

[0016] (4) Under the catalysis of the organic nitrogen base DIPEA, compound III reacts with the chlorinating reagent phosphorus oxychloride to obtain compound IV;

[0017] (5) Compound IV reacts with hydrazine monohydrate to obtain compound V;

[0018] (6) Under the condition of high-temperature reflux, compound V undergoes an intramolecular cyclization reaction to obtain compound VI;

[0019] (7) Finally, part of compound VI is deprotected or undergoes a substitution reaction to obtain the quinazolinone-fused triazole ring derivative I; The synthetic route is as follows:

[0020]

[0021] wherein R1 is n is 0 to 6, X is H or a 5- to 7-membered aliphatic heterocycle, the ring heteroatoms are N, O, Y is N, S or a halogen, L is H, a halogen or R2 is a halogen, an alkoxy group or a hydroxyl group.

[0022] Preferably, in step (1), the reaction solvent is selected from 1,2-dichloroethane, acetonitrile, N,N-dimethylformamide, dimethyl sulfoxide or toluene; the reaction temperature is 70 °C to 100 °C.

[0023] Preferably, in step (2), the reaction solvent is selected from 1,2-dichloroethane, N,N-dimethylformamide, acetonitrile, acetone or toluene; the reaction temperature is 20 °C to 35 °C.

[0024] Preferably, in step (3), the reaction solvent is selected from one or more of tetrahydrofuran, N,N-dimethylformamide, acetonitrile, ethyl acetate, toluene or acetone; the reaction temperature is -40 °C to -80 °C.

[0025] Preferably, in step (4), the reduction reaction solvent is selected from one or more of acetonitrile, 1,2-dichloroethane, chloroform, tetrahydrofuran, toluene, N,N-dimethylformamide, dimethyl sulfoxide; the reaction temperature is 90 °C to 120 °C.

[0026] Preferably, in step (5), the reaction solvent is selected from one or more of short-chain fatty acids, ethanol, dichloromethane, pyridine, acetonitrile, polyphosphoric acid, chloroform, acetone, tetrahydrofuran, N,N-dimethylformamide or dioxane; the reaction reagent is selected from bromoacetonitrile, oxalyl chloride, carbon disulfide or α-nitroarylacetone; reflux reaction.

[0027] Preferably, in step (6) and / or (7), the reaction satisfies any one of the following conditions: the reaction solvent is selected from one or more of dichloromethane, dioxane, tetrahydrofuran, methanol or ethyl acetate; the acid is selected from trifluoroacetic acid, hydrochloric acid; the base is selected from potassium hydroxide, triethylamine or cesium carbonate; the catalyst is selected from potassium iodide, sodium iodide or copper(I) iodide; the reaction temperature is 20 °C to 35 °C.

[0028] Use of the quinazolinone-fused triazole ring derivatives of the present invention in the preparation of a medicament for treating cancer, wherein the cancer is a cancer with overexpression of MAT2A.

[0029] Beneficial effects: Compared with the prior art, the present invention has the following remarkable advantages: (1) The quinazolinone-fused triazole ring derivatives are modified for R 1 and R 2 to synthesize novel compounds targeting homodimers, which have obvious inhibitory effects on liver tumor cells. When acting on the liver cancer cell line HepG2, its IC 50 is as low as 0.34 μM at minimum; (2) The preparation method designs and synthesizes new quinazolinone-fused triazole ring derivatives, and in vitro studies have found that they can significantly inhibit the occurrence and development of liver cancer cells. Detailed implementation manners

[0030] Example 1

[0031] The quinazolinone-fused triazole ring derivative of the present invention has the chemical name of 8-chloro-6-phenyl-[1,2,4]triazolo[4,3-c]quinazolin-5(6H)-one (I1), and the synthesis route is as follows:

[0032]

[0033] (1)-(2) Synthesis of intermediate II

[0034] Oxalyl chloride (3.22 mL, 38.02 mmol) was added to a solution of 4-chloro-2-fluorobenzamide (6.0 g, 57.6 mmol) in dichloroethane (120 mL) at room temperature. Then the reaction mixture was stirred at 80 °C for 30 minutes, and the reaction was monitored by TLC until completion. After the reaction mixture was cooled to room temperature, aniline (6.31 mL, 69.14 mmol) was added to the reaction solution. The reaction solution continued to react at room temperature for 1 hour. The reaction mixture was cooled with ice water and then slowly poured into a saturated aqueous sodium bicarbonate solution. Then it was stirred for 10 minutes. The precipitate was filtered and collected, dried under vacuum, and purified by silica gel column chromatography to obtain intermediate II. The yield was 65%, and it was a white solid; 1 1H NMR (400 MHz, DMSO-d6) δ 11.14 (s, 1H, CONHCO), 10.40 (s, 1H, Ar-NH-CO), 7.74 (t, J = 8.0 Hz, 1H, Ar-H), 7.65 (dd, J = 10.3, 2.0 Hz, 1H, Ar-H), 7.60 - 7.55 (m, 2H, Ar-H), 7.46 (dd, J = 8.4, 2.0 Hz, 1H, Ar-H), 7.40 - 7.33 (m, 2H, Ar-H), 7.16 - 7.09 (m, 1H, Ar-H). MS (ESI) m / z 293.1 [M + H]+ .

[0035] (3) Synthesis of Intermediate III

[0036] Dissolve Intermediate II (16.8 g, 43.0 mmol) in anhydrous THF solution (100 mL), and then slowly add it dropwise to a solution of KHMDS (126 mL, 126 mmol) under a nitrogen atmosphere. Stir at -78 °C for 1 hour, and then restore to room temperature and stir for 3.5 hours. Quench the reaction with saturated aqueous ammonium chloride solution to obtain a solid precipitate. Then filter and wash with water to obtain the crude product. Stir the crude product in water for 0.5 hours, then filter, wash with water, and dry in vacuo at 50 °C to obtain Intermediate III. Yield 62%, light yellow solid; 1 H NMR (400 MHz, DMSO-d6) δ 11.82 (s, 1H, CONHCO), 8.04 (d, J = 8.4 Hz, 1H, Ar-H), 7.66 - 7.56 (m, 3H, Ar-H), 7.49 - 7.43 (m, 2H, Ar-H), 7.32 (dd, J = 8.4, 1.9 Hz, 1H, Ar-H), 6.32 (d, J = 1.9 Hz, 1H, Ar-H). MS (ESI) m / z 273.0 [M+H] + .

[0037] (4) Synthesis of Intermediate IV

[0038] Add N,N-diisopropylethylamine (4.28 mL, 24.57 mmol) and phosphorus oxychloride (1.12 ml, 12.29 mmol) to a solution of Intermediate III1 (1.34 g, 4.91 mmol) in acetonitrile (0.1 M, 50 mL) at 0 °C. Stir at 100 °C for 4 hours, and monitor the end of the reaction by TLC. Cool the reaction solution to room temperature, pour it into ice water, extract with ethyl acetate, separate the organic phase, wash with water and saturated brine, then dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure at 10 °C to obtain Intermediate IV. Yield 95%, brown solid, purity 97.05%; MS (ESI) m / z 291.0 [M+H] + .

[0039] (5) Synthesis of Intermediate V

[0040] Intermediate IV (1.43 g, 4.91 mmol) was dissolved in anhydrous ethanol (20.0 mL, 4.91 mmol) solution. Hydrazine monohydrate (2.38 mL, 49.12 mmol) was slowly added dropwise at 4 °C, and the reaction was continuously stirred for 1 hour. Then it was restored to room temperature and stirred for another 1 hour. The reaction was monitored by TLC until completion. It was filtered and washed with water 2 - 3 times to obtain Intermediate V. The yield was 79%, off-white solid, purity 95.76%, m.p. 335.9 - 337.1 °C; 1 H NMR (400 MHz, DMSO-d6) δ 7.83 (d, J = 8.6 Hz, 1H, Ar-H), 7.65 - 7.57 (m, 2H, Ar-H), 7.56 - 7.49 (m, 1H, Ar-H), 7.43 - 7.36 (m, 2H, Ar-H), 7.15 - 7.01 (m, 1H, Ar-H), 6.09 (s, 1H, Ar-H). MS (ESI) m / z 287.1 [M+H] + ; HRMS (ESI) calcd for C 14 H 11 ClN4O [M+H] + 287.0694, found 287.0699.

[0041] (6) Synthesis of the target product I1

[0042] Intermediate V (100 mg, 348.77 μmol) was stirred with formic acid (1.5 mL) at 120 °C for 2 hours. The reaction was monitored by TLC until completion. Then the reaction solution was poured into ice water, filtered, washed with water, dried under reduced pressure, and purified by silica gel column chromatography to obtain product I1. The yield was 85%, light yellow solid, purity 100.00%, m.p. 275.9 - 276.9 °C; 1 H NMR (400 MHz, DMSO-d6) δ 9.47 (s, 1H, N=CH), 8.37 (d, J = 8.4 Hz, 1H, Ar-H), 7.75 - 7.62 (m, 3H, Ar-H), 7.54 (ddd, J = 18.1, 8.5, 1.8 Hz, 3H, Ar-H), 6.45 (d, J = 1.9 Hz, 1H, Ar-H). 13 C NMR (101 MHz, DMSO-d6) δ 147.4, 144.0, 140.2, 139.5, 136.6, 136.1, 131.0, 130.4, 129.6, 126.0, 124.7, 116.2, 109.9. MS (ESI) m / z 297.1 [M+H] + ; HRMS (ESI) calcd for C 15H9ClN4O[M+H] + 297.0538, found 297.0536.

[0043] Example 2

[0044] The quinazolinone-fused triazole ring derivatives of the present invention, with the chemical name 8-chloro-3-methyl-6-phenyl-[1,2,4]triazolo[4,3-c]quinazolin-5(6H)-one (I2), have the following synthetic route:

[0045]

[0046] Steps (1)-(5) are the same as in Example 1.

[0047] On the basis of step (6) of Example 1, formic acid was replaced with acetic acid, and other conditions remained unchanged to obtain the target product I2. Intermediate V (100 mg, 348.77 μmol) was stirred with acetic acid (1.5 mL) at 120 °C for 2 hours, and the reaction was monitored by TLC until completion. Then the reaction mixture was poured into ice water, filtered, washed with water, dried under reduced pressure, and purified by silica gel column chromatography to obtain product I2. The yield was 80%, a white solid, with a purity of 99.26%, m.p. 303.9 - 305.3 °C; 1 H NMR (400 MHz, DMSO-d6) δ 8.29 (d, J = 8.4 Hz, 1H, Ar-H), 7.73 - 7.61 (m, 3H, Ar-H), 7.56 - 7.51 (m, 2H, Ar-H), 7.46 (dd, J = 8.4, 1.9 Hz, 1H, Ar-H), 6.37 (d, J = 1.9 Hz, 1H, Ar-H), 2.77 (s, 3H, N = CCH3). 13 C NMR (101 MHz, DMSO-d6) δ 145.0, 148.1, 145.3, 140.3, 136.3, 136.2, 130.9 130.3, 129.6, 125.7, 124.5, 116.0, 110.3, 13.5. MS(ESI) m / z 311.1 [M+H] + ; HRMS(ESI) calcd for C 16 H 11 ClN4O [M+H] + 311.0694, found 311.0696.

[0048] Example 3

[0049] The quinazolinone-fused triazole ring derivatives of the present invention, with the chemical name 8-chloro-3-ethyl-6-phenyl-[1,2,4]triazolo[4,3-c]quinazolin-5(6H)-one (I3), have the following synthetic route:

[0050]

[0051] Steps (1) to (5) are the same as in Example 1.

[0052] On the basis of step (6) of Example 1, formic acid was replaced with butyric acid, and other conditions remained unchanged to obtain the target product I3. The yield was 70%, a light yellow solid, the purity was 99.15%, m.p. 234.9 - 236.1 °C; 1 H NMR (300 MHz, Chloroform-d) δ 8.42 (d, J = 8.4 Hz, 1H, Ar-H), 7.76 - 7.60 (m, 3H, Ar-H), 7.42 - 7.31 (m, 3H, Ar-H), 6.62 (d, J = 1.8 Hz, 1H, Ar-H), 3.35 (q, J = 7.5 Hz, 2H, N = CCH2), 1.47 (t, J = 7.5 Hz, 3H, CH3). 13 C NMR (75 MHz, Chloroform-d) δ 139.1, 137.9, 135.2, 130.9, 130.3, 128.8, 125.8, 125.1, 116.5, 109.5, 29.7, 20.9, 12.1. MS (ESI) m / z 325.1 [M + H] + ; HRMS (ESI) calcd for C 17 H 13 ClN4O [M + H] + 325.0851, found 325.0851.

[0053] Example 4

[0054] The quinazolinone-fused triazole ring derivatives of the present invention, with the chemical name 8-chloro-6-phenyl-3-propyl-[1,2,4]triazolo[4,3-c]quinazolin-5(6H)-one (I4), have the following synthetic route:

[0055]

[0056] On the basis of step (6) of Example 1, formic acid was replaced with valeric acid, and other conditions remained unchanged to obtain the target product I4. The yield was 64%, a white solid, the purity was 97.28%, m.p. 197.9 - 199.0 °C; 11H NMR (300 MHz, Chloroform-d) δ 8.42 (d, J = 8.4 Hz, 1H, Ar-H), 7.76 - 7.60 (m, 3H, Ar-H), 7.37 (ddd, J = 10.6, 8.3, 1.9 Hz, 3H, Ar-H), 6.62 (d, J = 1.9 Hz, 1H, Ar-H), 3.35 - 3.21 (m, 2H, N=CCH2), 1.91 (h, J = 7.4 Hz, 2H, CH2), 1.06 (t, J = 7.4 Hz, 3H, CH3). 13 13C NMR (75 MHz, Chloroform-d) δ 154.0, 139.1, 137.9, 135.2, 130.9, 130.3, 128.8, 125.8, 125.1, 116.5, 109.5, 29.7, 29.0, 21.4, 13.9. MS (ESI) m / z 339.1 [M+H] + ; HRMS (ESI) calcd for C 18 H 15 ClN4O [M+H] + 339.1007, found 339.1009.

[0057] Example 5:

[0058] The quinazolinone-fused triazole ring derivative of the present invention, with the chemical name 3-amino-8-chloro-6-phenyl-[1,2,4]triazolo[4,3-c]quinazolin-5(6H)-one (I5), has the following synthetic route:

[0059]

[0060] Based on the step (5) of Example 1, intermediate V (80 mg, 279.02 μmol) and bromoacetonitrile (29.55 mg, 279.02 μmol) were dissolved in ethanol (5 mL) solution, stirred at 100 °C for 2 hours, and the reaction was monitored by TLC until completion. Then the reaction solution was poured into ice water, filtered, washed with water, dried under reduced pressure, and recrystallized from ethanol to obtain product I5. Yield 91%, white solid, purity 99.67%, m.p. 336.3 - 338.1 °C; 1 1H NMR (400 MHz, DMSO-d6) δ 8.10 (d, J = 8.4 Hz, 1H, Ar-H), 7.72 - 7.58 (m, 3H, Ar-H), 7.57 - 7.51 (m, 2H, Ar-H), 7.38 (dd, J = 8.4, 2.0 Hz, 1H, Ar-H), 6.79 (s, 2H, N=CNH2), 6.29 (d, J = 2.0 Hz, 1H, Ar-H).13 C NMR(101MHz, DMSO-d6) δ 153.3, 146.4, 142.5, 139.9, 136.0, 134.9, 130.9, 130.2, 129.8, 124.6, 124.4, 116.0, 111.3. MS(ESI) m / z 311.9 [M+H] + ; HRMS(ESI) calcd for C 15 H 10 ClN5O [M+H] + 312.0647, found 312.0648.

[0061] Example 6:

[0062] The quinazolinone-fused triazole ring derivative of the present invention, with the chemical name of N-(8-chloro-5-oxo-6-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-c]quinazolin-3-yl)acetamide (I6), has the following synthetic route:

[0063]

[0064] Based on Example 5, the product I5 (50.0 mg, 160.40 μmol) was added dropwise to a solution of oxalyl chloride (17.2 μL, 240.59 μmol) and triethylamine (33.4 μL, 240.59 μmol) in dichloromethane (1.5 mL), and stirred at room temperature for 1.5 hours. The reaction was monitored by TLC until completion. Then, it was diluted with water, washed with saturated sodium bicarbonate and saturated brine respectively, the organic phase was separated, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the product I6. The yield was 72%, white solid, purity 95.15%, m.p. 254.5 - 256.3 °C; 1 H NMR(300MHz, DMSO-d6) δ 10.61(s, 1H, CONH), 8.31(d, J = 8.4Hz, 1H, Ar-H), 7.68(qd, J = 7.6, 6.5, 3.6Hz, 3H, Ar-H), 7.51(ddd, J = 10.3, 8.3, 1.9Hz, 3H, Ar-H), 6.38(d, J = 1.9Hz, 1H, Ar-H), 2.09(s, 3H, COCH3). 13 C NMR(75MHz, DMSO-d6) δ 170.4, 147.4, 145.1, 144.1, 140.3 136.5, 136.0, 131.0, 130.4, 129.6, 125.5, 124.7, 116.1, 110.1, 23.2. MS(ESI) m / z 354.0 [M+H]+ ; HRMS(ESI) calcd for C 17 H 12 ClN5O2 [M+H] + 354.0752, found 354.0754.

[0065] Example 7

[0066] The quinazolinone-fused triazole ring derivative of the present invention, with the chemical name of 8-chloro-3-mercapto-6-phenyl-[1,2,4]triazolo[4,3-c]quinazolin-5(6H)-one (I7), has the following synthetic route:

[0067]

[0068] On the basis of step (5) of Example 1, intermediate V (120.0 mg, 418.53 μmol) and carbon disulfide (31.9 mg, 418.53 μmol) were added to a pyridine (4 mL) solution, stirred at 120 °C for 2.5 hours, and the reaction was monitored by TLC until completion. After cooling to room temperature, the reaction solution was poured into ice water, and the crude product was obtained by suction filtration. The product I7 was obtained by silica gel column chromatography. The yield was 61%, a white solid, with a purity of 96.31%, m.p. 310.9 - 313.1 °C; 1 H NMR (300 MHz, DMSO-d6) δ 14.38 (s, 1H, N=C-SH), 8.07 (d, J = 8.4 Hz, 1H, Ar-H), 7.65 (dq, J = 14.3, 6.9 Hz, 3H, Ar-H), 7.54 - 7.45 (m, 2H, Ar-H), 7.37 (dd, J = 8.4, 2.0 Hz, 1H, Ar-H), 6.24 (d, J = 1.9 Hz, 1H, Ar-H). 13 C NMR (75 MHz, DMSO-d6) δ 165.7, 145.5, 144.7, 141.4, 136.8, 136.4, 131.0, 130.2, 129.6, 125.6, 124.1, 115.7, 109.7. MS(ESI) m / z 329.0 [M+H] + ; HRMS(ESI) calcd for C 15 H9ClN4OS [M+H] + 329.0258, found 329.0259.

[0069] Example 8

[0070] The quinazolinone-fused triazole ring derivatives of the present invention, with the chemical name 8-chloro-3-(methylthio)-6-phenyl-[1,2,4]triazolo[4,3-c]quinazolin-5(6H)-one (I8), have the following synthetic route:

[0071]

[0072] Based on Example 7, the product I7 (30.0 mg, 91.25 μmol) and potassium hydroxide (5.12 mg, 91.25 μmol) were added to a methanol (1.5 mL) solution, and then dimethyl sulfate (8.64 μL, 91.25 μmol) was added dropwise. The mixture was stirred at room temperature for 5 hours, and the reaction was monitored by TLC until completion. After cooling to room temperature, the reaction solution was poured into ice water, and the crude product was obtained by filtration. The target product I8 was obtained by silica gel column chromatography. Yield: 82%, white solid, purity: 98.71%, m.p. 253.0 - 254.7 °C; 1 H NMR (400 MHz, Chloroform-d) δ 8.31 (d, J = 8.5 Hz, 1H, Ar-H), 7.74 - 7.61 (m, 3H, Ar-H), 7.39 (dt, J = 8.5, 1.9 Hz, 3H, Ar-H), 6.71 (d, J = 1.8 Hz, 1H, Ar-H), 2.79 (s, 3H, SCH3). MS (ESI) m / z 343.0 [M+H] + ; HRMS (ESI) calcd for C 16 H 11 ClN4OS [M+H] + 343.0415, found 343.0416.

[0073] Example 9

[0074] The quinazolinone-fused triazole ring derivatives of the present invention, with the chemical name 8-chloro-3,6-diphenyl-[1,2,4]triazolo[4,3-c]quinazolin-5(6H)-one (I9), have the following synthetic route:

[0075]

[0076] On the basis of step (5) of Example 1, polyphosphoric acid (1.5 mL) and α-nitroacetophenone (5.12 mg, 91.25 μmol) were stirred at 130 °C, and then Intermediate V (30.0 mg, 91.25 μmol) was added in batches. Stirring was continued at 130 °C for 5 hours, and the reaction was monitored by TLC until completion. After cooling to room temperature, the reaction was quenched with ammonia water (25%), extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The product I9 was obtained by silica gel column chromatography. The yield was 85%, a light yellow solid, with a purity of 100.00%, m.p. 236.5 - 236.9 °C; 1 H NMR (300 MHz, Chloroform-d) δ 8.55 (d, J = 8.4 Hz, 1H, Ar-H), 7.88 - 7.79 (m, 2H, Ar-H), 7.70 - 7.59 (m, 3H, Ar-H), 7.52 - 7.35 (m, 6H, Ar-H), 6.66 (d, J = 1.8 Hz, 1H, Ar-H). 13 C NMR (75 MHz, Chloroform-d) δ 152.7, 148.4, 144.3, 139.0, 138.3, 135.3, 130.8, 130.6, 130.4, 130.3, 128.8, 127.9, 126.5, 126.0, 125.2, 116.5, 109.3. MS (ESI) m / z 372.9 [M+H] + ; HRMS (ESI) calcd for C 21 H 13 ClN4O [M+H] + 373.0851, found 373.0853.

[0077] Example 10

[0078] The quinazolinone-fused triazole ring derivative of the present invention, with the chemical name 8-chloro-3-(chloromethyl)-6-phenyl-[1,2,4]triazolo[4,3-c]quinazolin-5(6H)-one (I 10 ), and its synthetic route is shown as follows:

[0079]

[0080] On the basis of step (5) of Example 1, Intermediate V (100 mg, 348.77 μmol) and chloroacetyl chloride (27.76 μL, 348.77 μmol) were dissolved in glacial acetic acid (1.2 mL), and the mixture was refluxed at 120 °C for 3 hours. The reaction was monitored by LC-MS until completion, and the excess acetic acid was removed by distillation under reduced pressure. The target product I was obtained by silica gel column chromatography. 10Yield: 74%, white solid, purity: 95.60%, m.p. 256.7 - 257.9 °C; 1 1H NMR (400 MHz, Chloroform-d) δ 8.48 (d, J = 8.4 Hz, 1H, Ar-H), 7.76 - 7.62 (m, 3H, Ar-H), 7.44 - 7.38 (m, 3H, Ar-H), 6.69 (d, J = 1.8 Hz, 1H, Ar-H), 5.21 (s, 2H, CH2Cl). MS (ESI) m / z 345.0 [M+H] + ; HRMS (ESI) calcd for C 16 H 10 Cl2N4O [M+H] + 345.0304, found 345.0305.

[0081] Example 11

[0082] The quinazolinone-fused triazole ring derivative of the present invention, with the chemical name 8-chloro-3-(morpholinomethyl)-6-phenyl-[1,2,4]triazolo[4,3-c]quinazolin-5(6H)-one (I 11 ), and its synthetic route is as follows:

[0083]

[0084] Based on Example 10, the product I 10 (60.0 mg, 173.82 μmol) and potassium iodide (28.5 mg, 173.82 μmol) were dissolved in tetrahydrofuran (1.2 mL) solution, then morpholine (1.5 eq, 260.73 μmol) was added, and the reaction was stirred overnight at room temperature. The reaction was monitored by TLC until completion. Then, it was evaporated to dryness under reduced pressure, washed with saturated brine, extracted with ethyl acetate, the organic phase was separated, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the target product I 11 . Yield: 62%, off-white solid, purity: 96.08%, m.p. 139.9 - 141.1 °C; 1 1H NMR (400 MHz, Chloroform-d) δ 8.42 (d, J = 8.4 Hz, 1H, Ar-H), 7.72 - 7.59 (m, 3H, Ar-H), 7.41 - 7.37 (m, 2H, Ar-H), 7.34 (dd, J = 8.5, 1.9 Hz, 1H, Ar-H), 6.60 (d, J = 1.8 Hz, 1H, Ar-H), 4.19 (s, 2H, N = CCH2N), 2.66 (t, J = 5.3 Hz, 4H, O(CH2)2), 1.57 (q, J = 5.7 Hz, 4H, N(CH2)2).13 13C NMR (101 MHz, Chloroform-d) δ 150.4, 148.1, 144.7, 139.2, 138.0, 135.2, 130.9, 130.3, 128.8, 125.8, 125.1, 116.5, 109.3, 54.5, 53.2, 25.8, 24.0. MS (ESI) m / z 394.1 [M+H] + ; HRMS (ESI) calcd for C 20 H 18 ClN5O2 [M+H] + 396.1222, found 396.1257.

[0085] Example 12

[0086] The quinazolinone-fused triazole ring derivative of the present invention, with the chemical name 8-chloro-6-phenyl-3-(piperidin-1-ylmethyl)-[1,2,4]triazolo[4,3-c]quinazolin-5(6H)-one (I 12 ), and its synthetic route is as follows:

[0087]

[0088] Based on Example 11, piperidine was used to replace morpholine, and other conditions remained unchanged to obtain the target product I 12 . The yield was 75%, a light yellow solid, with a purity of 99.05%, m.p. 259.7 - 260.3 °C; 1 1H NMR (400 MHz, Chloroform-d) δ 8.45 (d, J = 8.4 Hz, 1H, Ar-H), 7.75 - 7.62 (m, 3H, Ar-H), 7.44 - 7.34 (m, 3H, Ar-H), 6.63 (d, J = 1.8 Hz, 1H, Ar-H), 4.25 (s, 2H, N = CCH2N), 3.73 (t, J = 4.7 Hz, 4H, O(CH2)2), 2.75 (s, 4H, (CH2)2). 13 13C NMR (75 MHz, Chloroform-d) δ 144.68, 139.24, 138.20, 135.15, 130.92, 130.38, 128.79, 125.89, 125.19, 116.58, 109.26, 66.77, 53.50, 52.96. MS (ESI) m / z 396.0 [M+H] + ; HRMS (ESI) calcd for C 21 H 20 ClN5O [M+H]+ 394.1429, found 394.1433.

[0089] Example 13

[0090] The quinazolinone-fused triazole ring derivative of the present invention, with the chemical name 8-chloro-6-phenyl-3-(pyrrolidin-1-ylmethyl)-[1,2,4]triazolo[4,3-c]quinazolin-5(6H)-one (I 13 ), and its synthetic route is as follows:

[0091]

[0092] Based on Example 11, morpholine was replaced with pyrrolidine, and other conditions remained unchanged to obtain the target product I 13 . The yield was 62%, a yellow solid, with a purity of 99.05%, m.p. 171.5 - 173.1 °C; 1 H NMR (300 MHz, Chloroform-d) δ 8.43 (dd, J = 8.5, 2.4 Hz, 1H, Ar-H), 7.72 - 7.61 (m, 3H, Ar-H), 7.42 - 7.34 (m, 3H, Ar-H), 6.61 (d, J = 1.8 Hz, 1H, Ar-H), 4.41 (s, 2H, N = CCH2N), 2.86 - 2.77 (m, 4H, N(CH2)2), 1.83 (q, J = 3.4 Hz, 4H, CH2CH2). 13 C NMR (75 MHz, Chloroform-d) δ 148.1, 144.8, 139.2, 138.1, 135.2, 130.8, 130.3, 128.8, 125.9, 125.1, 116.5, 109.3, 54.3, 50.5, 23.6. MS(ESI) m / z 380.1 [M+H] + ; HRMS(ESI) calcd for C 20 H 18 ClN5O [M+H] + 380.1273, found 380.1274.

[0093] Example 14

[0094] The quinazolinone-fused triazole ring derivative of the present invention, with the chemical name 8-chloro-6-phenyl-3-(piperazin-1-ylmethyl)-[1,2,4]triazolo[4,3-c]quinazolin-5(6H)-one (I 14 ), and its synthetic route is as follows:

[0095]

[0096] On the basis of Example 11, tert-butyl piperazine-1-carboxylate was used to replace morpholine, and other conditions remained unchanged to obtain the crude product. Then, it was dissolved in dichloromethane (2 mL) solution with trifluoroacetic acid (1.5 eq), and stirred at room temperature for 2 hours. The reaction was monitored by TLC until completion. The reaction solution was diluted with water, washed with saturated aqueous sodium bicarbonate and saturated brine respectively, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the target product I. 14 . The yield was 57%, a yellow solid, with a purity of 99.05%, m.p. 175.3 - 176.8 °C; 1 1H NMR (300 MHz, Chloroform-d) δ 8.43 (d, J = 8.4 Hz, 1H, Ar-H), 7.76 - 7.61 (m, 3H, Ar-H), 7.44 - 7.32 (m, 3H, Ar-H), 6.62 (d, J = 1.8 Hz, 1H, Ar-H), 4.24 (s, 2H, N = CCH2N), 3.20 - 3.03 (m, 4H, N(CH2)2), 2.92 (t, J = 4.9 Hz, 4H, N(CH2)2'), 1.99 (s, 1H, NH). 13 13C NMR (75 MHz, Chloroform-d) δ 149.5, 148.4, 144.6, 139.2, 138.3, 135.1, 131.0, 130.4, 128.7, 125.9, 125.3, 116.6, 109.2, 52.4, 50.8, 44.2, 29.7. MS (ESI) m / z 396.0 [M + H] + ; HRMS (ESI) calcd for C 20 H 19 ClN6O [M + H] + 395.1382, found 395.1390.

[0097] Example 15

[0098] The quinazolinone-fused triazole ring derivative of the present invention, with the chemical name 3,8-dichloro-6-phenyl-[1,2,4]triazolo[4,3-c]quinazolin-5(6H)-one (I 15 ), and its synthetic route is shown as follows:

[0099]

[0100] On the basis of Example 1, 1-chloropyrrolidine-2,5-dione (33.75 mg, 252.77 μmol) was added to a chloroform solution of Product I (50.00 mg, 168.51 μmol), and the reaction was stirred overnight at 75 °C. The reaction was monitored by TLC until completion. Then, the solvent was evaporated under reduced pressure, washed with saturated brine, extracted with ethyl acetate, the organic phase was separated, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the target product I was obtained by silica gel column chromatography. 15 . The yield was 65%, a light beige solid, the purity was 99.05%, m.p. 238.1 - 239.4 °C; 1 H NMR (300 MHz, Chloroform-d) δ 8.41 (d, J = 8.4 Hz, 1H, Ar-H), 7.77 - 7.60 (m, 3H, Ar-H), 7.39 (ddd, J = 7.6, 3.5, 1.7 Hz, 3H, Ar-H), 6.65 (d, J = 1.8 Hz, 1H, Ar-H). 13 C NMR (75 MHz, Chloroform-d) δ 153.9, 145.5, 143.5, 139.3, 138.7, 134.7, 131.0, 130.5, 128.7, 125.6, 125.5, 116.8. MS (ESI) m / z 331.0 [M+H] + ; HRMS (ESI) calcd for C 15 H8Cl2N4NaO [M+Na] + 352.9967, found 352.2785.

[0101] (1) Enzyme activity assay of Compound I

[0102] The enzyme protein was diluted to 0.5 μM in buffer (50 mM Tris, pH 8.0, 50 mM KCl, 15 mM MgCl2, 0.3 mM EDTA, 0.005% [w / v] BSA). The test compound was prepared at 50× the desired final concentration in 100% DMSO. 1 μL volume of the compound dilution was added to 40 μL of the enzyme dilution, and the mixture was equilibrated at 25 °C for 60 minutes. Then, 10 μL of the substrate mixture (500 μM ATP, pH 7.0, 400 μM L-methionine in 1× buffer) was added for enzyme activity assay, and the mixture was incubated at 25 °C for another 60 minutes. The reaction was stopped (e.g., by adding cold deionized water to stop the reaction and dilute the sample), and the amount of released phosphate released by the enzyme through the production of SAM was measured using the PiColorLock kit. The absorbance (OD value) of the transparent microplate was measured at a wavelength of 620 nm on a microplate reader to calculate the inhibition rate, and the dose-response relationship graph was plotted by Graphpad prism to obtain the IC50 The inhibition rate is calculated by the following formula:

[0103] Inhibition rate = [(As - A0) - (At - A0)] / (As - A0) × 100%; where A0, At, and As represent the absorbance of the phosphate kit only, the test compound, and the control without the test compound, respectively.

[0104] Table 1 Inhibitory activity of compound I on MAT2A enzyme

[0105]

[0106]

[0107]

[0108] -- It means that there are no test results yet;

[0109] b Inhibition rate of the compound on HepG2 cells at a concentration of 50 nM;

[0110] c Concentration of 50% inhibitory effect of the compound on enzyme activity, and the data are the mean ± SD from at least 3 parallel experiments;

[0111] d Enzyme inhibition rate of the compound at a concentration of 3 μM.

[0112] Based on the drug design strategy of skeleton fusion technology, according to the drug structure optimization strategy of molecular hybridization and bioisosteres, the present invention has developed a series of novel inhibitor compounds I targeting MAT2A homodimer. In vitro studies have found that it can significantly inhibit the occurrence and development of cancer cells with high expression of MAT2A, such as liver cancer, etc.; it provides new ideas for the synthesis of new skeleton MAT2A homodimer inhibitors, and provides the possibility for the research and development of small molecule anti-tumor drugs targeting MAT2A homodimer, and has good application prospects.

[0113] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A quinazolinone-fused triazole ring derivative, characterized in that, The chemical structural formula is as follows: wherein R1 is n is from 0 to 6, X is H or a 5- to 7-membered aliphatic heterocycle; when X is a 5- to 7-membered aliphatic heterocycle, is being -Cl, -NH2, -SH, 2. The quinazolinone-fused triazole ring derivative according to claim 1, characterized in that, The said is H, 3. Use of a quinazolinone-fused triazole ring derivative according to any one of claims 1 to 2 in the preparation of an anti-tumor drug for treating overexpression of MAT2A.

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