Quinoline compounds and their applications

Abstract

The present invention relates to the field of pharmaceutical chemistry, and in particular to a class of quinoline compounds or pharmaceutically acceptable salts thereof, isomers thereof, solvates thereof, compositions thereof and medical uses thereof. Pharmacological experiments have shown that the quinoline compounds of the present invention have an effect on KRAS G12D At a concentration of 10 µM, compounds C1, C3, C4, C6, C11, C12, C13, C16, C22 and C23 of the present invention have good affinity for KRAS G12D ‑cRaf has a certain inhibitory effect. In particular, compounds C1 and C12 have a certain inhibitory effect on KRAS G12D ‑cRaf showed strong inhibitory effects.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry, and in particular to a class of quinoline compounds or pharmaceutically acceptable salts thereof, isomers thereof, solvates thereof, pharmaceutical compositions thereof and medical uses thereof. Background Art

[0002] RAS is one of the most frequently mutated oncogenes. It cycles between active (GTP-bound) and inactive (GDP-bound) states, acting as a molecular switch. RAS is a type of small GTPase that plays an important role in normal cell growth, development, proliferation, apoptosis, and cell signaling during movement. RAS activity is mainly regulated by guanine nucleotide exchange factors (GEFs) accelerating GDP dissociation to promote GTP binding, and GTPase activating proteins (GAPs) stimulating GTP hydrolysis to drive RAS to GDP binding. The three common RAS genes are KRAS, HRAS, and NRAS, of which KRAS mutations are the most common, mainly occurring in pancreatic cancer and lung cancer, accounting for about 85% of human cancers.

[0003] The KRAS molecule is activated when it binds to GTP and is inactivated when it binds to GDP. A variety of KRAS mutations lead to differential distribution of mutants in cancer. KRAS mutations cause RAS to bind to GTP, continuously activating downstream signaling pathways, leading to the occurrence and progression of tumors. The key to improving KRAS-driven cancers is to target the dysregulation of the interaction between KRAS mutants and effector proteins. The high incidence of KRAS mutations and its important role in many cancers make it a potential drug target. However, the surface of KRAS is smooth, there is no pocket for suitable inhibitor binding, and the binding affinity of KRAS to GTP / GDP reaches the picomolar level. Therefore, it is often considered an "undruggable target" and it is difficult to find inhibitors targeting the KRAS protein. Targeting KRAS G12C The launch of drugs AMG510 and MRTX849 has broken the concept that KRAS is "undruggable" and has also promoted the development of targeted KRAS G12D Research on inhibitors of KRAS G12D Mutations are predominant in pancreatic ductal adenocarcinoma and colorectal cancer. The small molecule binding site p110 in its structure is a potential allosteric regulatory site for targeting KRAS. G12D It provides a new strategy for the development of small molecule drugs.

[0004] Based on this research idea, the researchers discovered a new type of quinoline compound that has a positive effect on KARS. G12D It has good selectivity and anti-tumor cell proliferation activity. Summary of the invention

[0005] The technical problem solved by the present invention is to provide a new type of quinoline compound and a pharmaceutically acceptable salt thereof, a preparation method thereof, a pharmaceutical composition and an application thereof.

[0006] In order to solve the technical problem of the present invention, the present invention provides the following technical solutions:

[0007] The first aspect of the technical solution of the present invention is to provide a compound represented by the general formula (I) or a pharmaceutically acceptable salt, isomer or solvate thereof:

[0008]

[0009] Formula (I)

[0010] Where R is:

[0011]

[0012] Preferably, said R is: or .

[0013] The second aspect of the technical solution of the present invention is to provide a pharmaceutical composition comprising the compound described in the first aspect or its pharmaceutically acceptable salt, isomer, or solvate thereof, or its pharmaceutically acceptable salt and one or more pharmaceutical carriers and / or diluents, in any clinically or pharmaceutically acceptable dosage form, preferably an oral preparation or an injection. The pharmaceutical composition contains 0.01g to 10g of the compound represented by the general formula (I) in a physiologically effective amount, which can be 0.01g, 0.015g, 0.02g, 0.025g, 0.03g, 0.04g, 0.05g, 0.1g, 0.125g, 0.2g, 0.25g, 0.3g, 0.4g, 0.5g, 0.6g, 0.75g, 1g, 1.25g, 1.5g, 1.75g, 2g, 2.5g, 3g, 4g, 5g, 6g, 7g, 8g, 9g, 10g, etc.

[0014] Any compound of the present invention or a pharmaceutically acceptable salt, isomer or solvate thereof can be administered orally or parenterally to a patient in need of such treatment.

[0015] When used for parenteral administration, it can be made into an injection. When it is made into an injection, it can be produced by conventional methods in the existing pharmaceutical field, and aqueous solvents or non-aqueous solvents can be selected. The most commonly used aqueous solvent is water for injection, and 0.9% sodium chloride solution or other suitable aqueous solutions can also be used; the commonly used non-aqueous solvent is vegetable oil, mainly soybean oil for injection, and other aqueous solutions of ethanol, propylene glycol, polyethylene glycol, etc. When preparing an injection, no additives may be added, or appropriate additives may be added according to the properties of the drug, such as osmotic pressure regulators, pH regulators, solubilizers, fillers, antioxidants, antibacterial agents, emulsifiers, suspending agents, etc. When used for oral administration, it can be made into conventional solid preparations, such as tablets, capsules, pills, granules, etc.; it can also be made into oral liquid preparations, such as oral solutions, oral suspensions, syrups, etc. When making oral preparations, appropriate fillers, binders, disintegrants, lubricants, etc. may be added.

[0016] The compounds described in the present invention can be added with pharmaceutically acceptable carriers to prepare common pharmaceutical preparations, such as tablets, capsules, powders, syrups, liquids, suspensions, injections, and can be added with common pharmaceutical excipients such as flavors, sweeteners, liquid or solid fillers or diluents.

[0017] The clinical dosage of the compound of the present invention is 0.01-1000 mg / day, and may deviate from this range depending on the severity of the disease or the dosage form.

[0018] The third aspect of the technical solution of the present invention is to provide a method for preparing a drug for inhibiting KRASG12D activity in cells comprising the compound described in the first aspect or a pharmaceutically acceptable salt, isomer, or solvate thereof, and the pharmaceutical composition described in the second aspect.

[0019] The third aspect of the technical solution of the present invention is to provide a method for preparing a drug for treating KRAS G12D-related cancers comprising the compound described in the first aspect or a pharmaceutically acceptable salt, isomer, or solvate thereof and the pharmaceutical composition described in the second aspect.

[0020] The KRAS G12DThe relevant cancer is selected from the group consisting of angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma, myxoma, rhabdomyosarcoma, fibroma, lipoma, teratoma, lung cancer, bronchogenic carcinoma, alveolar carcinoma, bronchial adenoma, lymphoma, esophageal cancer, gastric cancer, pancreatic cancer, colorectal cancer, small intestine cancer, large intestine cancer, rectal cancer, kidney cancer, lymphoma, leukemia, bladder and urethra cancer, prostate cancer, testicular cancer, liver cancer, bile duct cancer, hepatoblastoma, hepatocellular adenoma, gallbladder cancer, bile duct cancer, bone cancer, fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, malignant lymphoma, multiple myeloma, malignant Giant cell tumors Chordoma, osteochondroma, benign enchondroma, chondroblastoma, chondromyxoid fibroma, glioma, germ cell tumor, glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors, spinal neurofibroma, endometrial carcinoma, malignant teratoma, vulvar squamous cell carcinoma, fallopian tube carcinoma, myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, multiple myeloma, malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi sarcoma, lipoma, hemangioma, cutaneous fibroma.

[0021] Preferably, the cancer is lung cancer, colorectal cancer or pancreatic cancer.

[0022] More preferably, the lung cancer is non-small cell lung cancer or small cell lung cancer.

[0023] Alternatively, more preferably, the cancer is pancreatic cancer.

[0024] Alternatively, more preferably, the cancer is colorectal cancer.

[0025] Beneficial technical effects:

[0026] The advantages of the present invention are:

[0027] (1) At a concentration of 10 μM, compounds C1, C3, C4, C6, C11, C12, C13, C16, C22 and C23 of the present invention have an inhibitory effect on KRAS G12D -cRaf has a certain inhibitory effect. In particular, compounds C1 and C12 have a certain inhibitory effect on KRAS G12D -cRaf showed strong inhibitory effects.

[0028] (2) Compounds with good single concentration inhibitory effects on KRAS were further tested. G12D -cRaf IC 50 Among them, C1 had the best activity, as low as 2.04 μM.

[0029] (3) The novel quinoline compounds designed and synthesized by the present invention have certain growth inhibition effects on tumor cells, IC 50The values ​​were basically around 10~30μM. Among them, C12 showed good anti-proliferative activity in 4 types of tumor cells, laying the foundation for the development of anti-cancer drugs. DETAILED DESCRIPTION

[0030] The present invention will be further described below in conjunction with specific implementation modes so that those skilled in the art can better understand the present invention, but the present invention is not limited thereto.

[0031] Unless otherwise specified, the reagents involved in the embodiments of the present invention are all commercially available products and can be purchased through commercial channels.

[0032] The compounds of the general formula (I) of the present invention can be prepared by the following method:

[0033]

[0034] R is:

[0035] .

[0036] Aniline (1) and dimethyl succinate were dissolved in methanol and subjected to substitution reaction under light-shielding conditions to generate aniline diester compound W1 with a high yield of 90%. Compound W1 was cyclized under the catalysis of PPA to generate quinolinone compound W2 with a yield of 50%. Compound W2 was catalyzed by NaBH in a mixed solvent of methanol and acetonitrile (1:1). 4 Reduction was performed to generate the key intermediate hydroxyquinoline compound W3 with a yield of 48%. Compound W3 underwent substitution reaction with an electrophilic reagent to obtain the target compound CR1.

[0037] Example 1 Synthesis of 2-(Hydroxymethyl)quinoline-4-fluorobenzenesulfonate (C1):

[0038]

[0039] Aniline (1) (42.7 mL, 469.0 mmol) was dissolved in methanol (50 mL), and dimethyl acetylenedicarboxylate (86.5 mL, 704.0 mmol) was added. The mixture was reacted at room temperature for 24 h. The raw material disappeared after TLC detection (developing solvent: ethyl acetate-petroleum ether = 1:10). The solvent was evaporated from the reaction solution under reduced pressure, and the crude product was purified by silica gel column chromatography (mobile phase: ethyl acetate-petroleum ether = 1:10) to obtain a yellow oily aniline quinoline compound W1. Synthesis of dimethyl 2-(phenylamino)maleate (W1) (25 g, 2.4 mmol) was added to PPA (100 g) and stirred at 25 oC, react for 3 h, and the raw material disappears when detected by TLC (developing solvent: ethyl acetate-petroleum ether = 1:1). The organic phase was washed with cold saturated sodium bicarbonate solution and ice water, filtered, and then some impurities were washed with ether. The filter cake was dried and the crude product was separated and purified by silica gel column chromatography (mobile phase: ethyl acetate-petroleum ether = 1:1) to obtain a white solid quinolinone compound W2. 4-Oxo-1,4-dihydroquinoline-2-carboxylic acid methyl ester (W2) (1 g, 4.9 mmol) was dissolved in MeOH (50 mL) and heated at 0 o C under the condition of adding NaBH 4 (1g, 24.6 mmoL), this reaction was carried out at 25 o C, react for 4 h. The organic phase was quenched with ice-saturated ammonium chloride aqueous solution, extracted with ethyl acetate, washed with saturated sodium chloride, dried with anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (mobile phase: methanol-dichloromethane = 1:15) to obtain a white solid hydroxyquinoline compound (W3). 2-(Hydroxymethyl)quinolin-4(1H)-one (W3) (1 mmol) was dissolved in acetonitrile (2 mL), anhydrous potassium carbonate was slowly added, and the electrophilic reagent 4-fluorobenzenesulfonyl chloride was slowly added under nitrogen protection, and the reaction was heated to reflux for 12 h. The reaction solution was quenched with water, extracted with dichloromethane, and the organic phase was washed with saturated brine, dried with anhydrous magnesium sulfate, filtered, and the solvent was evaporated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (mobile phase: dichloromethane: methanol = 30:1) to obtain the target compound (C1). The product is a white powdery solid, yield: 67%, mp: 85-87 o C. 1 H NMR (800 MHz, Methanol- d 4 ) δ 8.04 (m, 2H), 7.97 (d, J IR (KBr, cm -1): 554.65, 572.88, 691.89, 763.52, 772.76, 779.27, 842.77,874.24, 965.91, 1052.22, 1069.44, 1158.52, 1192.49, 1383.60, 1494.31,1508.26, 1590.12, 1601.07, 3212.10. HRMS (ESI): m / z Calculate for C 16 H 13 FNO 4 S + [M+H] + :334.0549, found: 334.0531.

[0040] The preparation methods of the compounds in Examples 2-10 are the same as those in Example 1, except that different electrophilic reagents are used. The specific electrophilic reagents used in each example are described in the corresponding example:

[0041] Example 2 Synthesis of 2-(hydroxymethyl)quinoline-4-(trifluoromethyl)benzenesulfonate (C3):

[0042]

[0043] The electrophilic reagent used is 4-trifluoromethylbenzenesulfonyl chloride. The product is a white powdery solid, yield: 67%, mp: 117-119 o C. 1 H NMR (800 MHz, DMSO-d6) δ 8.27 (d, J = 8.3 Hz, 1H), 8.11 (d,J = 8.4 Hz, 1H), 7.98 (t, J = 7.7 Hz, 1H), 7.80 (d, J = 8.1 Hz, 2H), 7.70 (d,J = 8.1 Hz, 3H), 6.94 (s, 1H), 4.85 (s, 2H). IR (KBr, cm -1):429.47, 558.83,604.12, 711.75, 731.70, 750.74, 768.97, 790.09, 852.42, 874.18, 888.40,968.26, 1014.66, 1045.56, HRMS (ESI): m / z Calculate for C 16 H 13 F 3 NO 4 S + [M+H] + : 384.0517, found: 384.0509.

[0044] Example 3 Synthesis of (4-(isopentyloxy)quinoline)methanol (C4):

[0045]

[0046] The electrophilic reagent used is 1-bromoisopentane. The product is a white powdery solid, yield: 67%, mp: 73-74 o C; 1 H NMR (800 MHz, DMSO- d 6 ) δ 8.12 – 8.08 (m, 1H), 7.87 (d, J = 8.3 Hz, 1H),7.72 – 7.68 (m, 1H), 7.53 – 7.49 (m, 1H), 7.14 (s, 1H), 5.53 (t, J = 5.9 Hz,1H), 4.68 – 4.65 (m, 2H), 4.29 (t, J = 6.5 Hz, 2H), 1.91 (p, J = 13.4, 6.7 Hz,1H), 1.79 (q, J = 6.6 Hz, 2H), 0.99 (d, J = 6.7 Hz, 6H). IR (KBr, cm -1): 770.98,853.06, 986.31, 1078.53, 1111.79, 1134.27, 1156.41, 1322.62, 1363.41,1429.16, 1471.23, 1508.98, 1569.80, 1590.85, 2870.13, 2931.57, 2956.46,3168.20. HRMS (ESI): m / z Calculate for C 15 H 20 NO 2 + [M+H] + : 246.1494, found: 246.1479.

[0047] Example 4 Synthesis of (4-(4,4,4-trifluorobutoxy)quinoline)methanol (C6):

[0048]

[0049] The electrophilic reagent used is 1-bromo-4,4,4-trifluorobutane. The product is a white powdery solid, yield: 67%, mp: 74-76 o C. 1 H NMR (800 MHz, DMSO- d 6 ) δ 8.17 (d, J = 8.2 Hz, 1H), 7.88 (d, J =8.4 Hz, 1H), 7.74 – 7.70 (m, 1H), 7.52 (t, J = 7.5 Hz, 1H), 7.12 (s, 1H), 5.56(t, J = 5.9 Hz, 1H), 4.69 – 4.65 (m, 2H), 4.33 (t, J = 6.1 Hz, 2H), 2.61 – 2.53(m, 2H), 2.15 – 2.09 (m, 2H). IR (KBr, cm -1): 616.00, 647.77, 765.69, 835.61,852.50, 981.74, 1012.20, 1048.42, 1083.89, 1113.60, 1150.23, 1192.94,1235.78, 1256.67, 1342.68, 1357.10, 1384.41, 1430.47, 1451.35, 1470.44,1513.54, 1574.17, 1599.64, 1620.25, 2924.98, 3133.89, 3404.36. HRMS (ESI): m / z Calculate for C 14 H 15 F 3 NO 2 + [M+H] + : 286.1055, found: 286.1043.

[0050] Example 5 Synthesis of (4-((3,4-dichlorophenyl)oxy)quinoline)methanol (C11):

[0051]

[0052] The electrophilic reagent used is 3,4-dichlorobenzyl chloride. The product is a white powdery solid, yield: 67%, mp: 120-123 o C. 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.17 (d, J = 8.2 Hz, 1H), 7.94 – 7.84 (m, 2H), 7.78 – 7.70 (m, 2H), 7.63 – 7.50 (m, 2H), 7.24 (s, 1H), 5.59 (t, J = 4.7 Hz,1H), 5.43 (s, 2H), 4.69 (d, J = 4.4 Hz, 2H). IR (KBr, cm -1): 661.51, 757.10,809.39, 977.30, 1030.54, 1078.70, 1113.85, 1133.77, 1157.65, 1235.17,1350.88, 1372.70, 1402.89, 1425.90, 1472.33, 1510.16, 1569.38, 1597.19,1621.51, 2909.49, 3140.71. HRMS (ESI): m / z Calculate for C 17 H 15 Cl 2 NO 2 + [M+H] + :334.0402, found: 334.0392.

[0053] Example 6 Synthesis of (4-((3,5-bis(trifluoromethyl)benzyl)oxy)quinoline)methanol (C12):

[0054]

[0055] The electrophilic reagent used is 3,5-bis(trifluoromethyl)benzyl chloride. The product is a white powdery solid, yield: 67%, mp: 124-125 o C. 1 H NMR (800 MHz, DMSO- d 6 ) δ 8.33 (s, 2H), 8.18 – 8.14 (m, 2H), 7.91 (d, J = 8.3 Hz, 1H), 7.74 (t, J = 7.6 Hz, 1H), 7.55 (t, J = 7.5 Hz, 1H), 7.29(s, 1H), 5.62 – 5.60 (m, 3H), 4.70 (d, J = 5.8 Hz, 2H). IR (KBr, cm -1): 683.39,703.71, 765.85, 777.88, 843.81, 890.20, 973.19, 1065.40, 1121.13, 1186.71,1204.75, 1237.95, 1280.30, 1349.80, 1370.87, 1430.20, 1468.74, 1512.46,1572.54, 1599.27, 1621.98, 3397.16. HRMS (ESI): m / z Calculate for C 19 H 14 F 6 NO 2 + [M+H] + :402.0929, found: 402.0889.

[0056] Example 7 Synthesis of (4-((3-fluorobenzyl)oxy)quinoline)methanol (C13):

[0057]

[0058] The electrophilic reagent used is m-fluorobenzyl chloride. The product is a white powdery solid, yield: 67%, mp: 116-118 o C. 1 H NMR (800 MHz, DMSO- d 6 ) δ 8.25 – 8.21 (m, 1H), 8.09 (d, J = 8.4 Hz, 1H),8.03 (s, 1H), 7.87 – 7.80 (m, 5H), 7.74 (s, 1H), 7.68 – 7.64 (m, 1H), 5.57(s, 2H), 3.93 (t, J = 5.9 Hz, 2H), 3.11 (s, 2H). IR (KBr, cm -1): 681.89, 760.98,788.20, 843.10, 863.29, 924.93, 978.05, 1023.67, 1042.66, 1085.14, 1116.93,1135.19, 1160.32, 1235.29, 1249.42, 1284.32, 1307.79, 1357.59, 1386.67,1424.87, 1458.62, 1492.14, 1509.84, 1572.29, 1597.58, 1619.43, 2851.69,2923.79, 3195.16. HRMS (ESI): m / z Calculate for C 17 H 15 FNO 2 + [M+H] + : 284.1087, found:284.1063.

[0059] Example 8 Synthesis of 2-(Hydroxymethyl)quinoline-3,4,5-trifluorobenzenesulfonate (C16):

[0060]

[0061] The electrophilic reagent used is 3,4,5-trifluorobenzenesulfonyl chloride. The product is a white powdery solid, yield: 67%, mp: 106-108 o C. 1 H NMR (800 MHz, DMSO- d 6 ) δ 8.30 – 8.26 (m, 1H), 8.13 (d, J = 8.5Hz, 1H), 8.01 – 7.97 (m, 1H), 7.71 (t, J = 7.6 Hz, 1H), 7.42 – 7.36 (m, 2H), 6.98 (s, 1H), 4.87 (s, 2H). IR (KBr, cm -1): 592.44, 619.10, 683.69, 761.96,774.53, 792.64, 865.18, 880.36, 922.29, 964.71, 1039.86, 1053.90, 1080.35,1178.93, 1327.89, 1351.83, 1370.69, 1420.99, 1441.22, 1506.77, 1524.45,1563.03, 1604.66, 1625.45, 3202.03. HRMS (ESI): m / z Calculate for C 16 H 11 F 3 NO 4 S + [M+H] + :370.0361, found: 370.0334.

[0062] Example 9 Synthesis of 2-(Hydroxymethyl)quinoline-4-cyanobenzenesulfonate (C22):

[0063]

[0064] The electrophilic reagent used is 4-cyanobenzenesulfonyl chloride. The product is a white powdery solid, yield: 57%, mp: 128-130 o C. 1 H NMR (800 MHz, DMSO- d 6 ) δ 8.30 (d, J = 8.3 Hz, 1H), 8.18 (d, J = 8.5Hz, 1H), 8.05 – 8.01 (m, 1H), 7.81 (d, J = 8.4 Hz, 2H), 7.75 (d, J = 8.3 Hz, 3H),7.09 – 7.06 (m, 1H), 4.90 (s, 2H). IR (KBr, cm -1): 517.32, 569.63, 635.71,678.39, 755.44, 773.52, 790.25, 812.11, 843.99, 873.45, 889.60, 958.22,966.32, 1016.10, 1035.75, 1068.12, 1090.28, 1147.07, 1180.55, 1190.72,1292.37, 1322.56, 1381.38, 1500.81, 1602.61, 2242.19, 2922.21, 3084.90,3105.65, 3525.70. HRMS (ESI): m / z Calculate for C 17 H 13 N 2 O 4 S + [M+H] + : 341.0596, found:341.0573.

[0065] Example 10 Synthesis of 2-(Hydroxymethyl)quinoline-4-bromobenzenesulfonate (C23):

[0066]

[0067] The electrophilic reagent used is 4-bromobenzenesulfonyl chloride. The product is a white powdery solid, yield: 73%, mp: 118-120 o C. 1 H NMR (800 MHz, DMSO- d 6 ) δ 8.28 (d, J = 8.2 Hz, 1H), 8.16 – 8.12 (m, 1H), 7.99 (dd, J = 14.9, 7.9 Hz, 1H), 7.73 – 7.69 (m, 1H), 7.54 (d, J = 8.4 Hz, 2H),7.53 – 7.49 (m, 2H), 6.99 (s, 1H), 4.87 (d, J = 8.0 Hz, 2H). IR (KBr, cm -1):421.72, 533.29, 563.42, 575.37, 604.69, 699.82, 735.05, 760.52, 775.46,828.73, 870.74, 890.78, 967.24, 1010.12, 1022.85, 1048.76, 1153.13, 1176.33,1191.10, 1335.23, 1384.98, 1410.77, 1469.18, 1503.48, 1560.86, 1570.39,1604.60, 1620.46, 2935.93, 3094.62, 3281.08. HRMS (ESI): m / z Calculate for C 16 H 13 BrNO 4 S + [M+H] + : 393.9749, found: 393.9728.

[0068] Some biological activity tests and results of the compounds of the present invention are as follows:

[0069] Experimental Example 1: Effects of the compounds of the present invention on KRAS G12D -cRaf affinity evaluation

[0070] 1.1. Experimental methods:

[0071] 0.1 μL of compound solution was transferred to a 384-well plate. The initial concentration of the test compound was 1 μM, and the test compound was diluted 3-fold to 10 concentrations. 2.5 μL of KRAS was added to each well. G12D , centrifuged at 1000 rpm for 1 min, added 2.5 μL cRAF to each well, centrifuged at 1000 rpm for 1 min, and incubated at 25°C for 15 min. Add 5 μL SA-Tb&GST-XL665 to the 384-well plate, centrifuged at 1000 rpm for 1 min, and incubated at 25°C for 60 min. HTRF signals (665 / 620 nm) were read using a BMG microplate reader. Inhibition rate or IC was calculated using GraphPadPrism software 50 value.

[0072] 1.2. Experimental results

[0073] 1. Single concentration inhibition rate results

[0074] Table 1 Effects of target compounds on KRAS G12D -cRaf affinity (10 μM)

[0075]

[0076] The preliminary screening results showed that at a concentration of 10 µM, compounds C1, C3, C4, C6, C11, C12, C13, C16, C22 and C23 of the present invention had an effective effect on KRAS G12D -cRaf has a certain inhibitory effect. In particular, compounds C1 and C12 have a certain inhibitory effect on KRAS G12D -cRaf showed strong inhibitory effects.

[0077] 2. Effects of CR series compounds on KRAS G12D -cRaf IC 50 value

[0078] Table 2 Effects of CR series compounds on KRAS G12D -cRaf IC 50 value

[0079]

[0080] The compounds with better single concentration inhibitory effects were further tested for KRAS G12D -cRaf IC 50 Among them, C1 had the best activity, as low as 2.04 μM.

[0081] Experimental Example 2: Determination of the inhibitory effect of target compounds on tumor cell growth

[0082] 2.1 MTT assay method

[0083] The compounds of the present invention are dissolved in dimethyl sulfoxide and prepared at 1 mg / mL in RPMI-1640 medium containing 10% fetal bovine serum for later use. When group administration is performed, the compounds are diluted to the required concentration by a certain multiple using the medium.

[0084] NCIH-1385 cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum and passaged every 2-3 days. The cell culture incubator was set at: CO 2 Concentration: 5%, Temperature: 37 o C, Cells in the logarithmic growth phase were used for the experiment.

[0085] PANC-1 cells were cultured in high-glucose DMEM medium containing 10% fetal bovine serum and passaged every 2-3 days. The cell culture incubator was set at: CO 2 Concentration: 5%, Temperature: 37 o C, Cells in the logarithmic growth phase were used for the experiment.

[0086] AsPc-1 cells were cultured in high-glucose DMEM medium containing 10% fetal bovine serum and passaged every 2-3 days. The cell culture incubator was set at: CO 2 Concentration: 5%, Temperature: 37 o C, Cells in the logarithmic growth phase were used for the experiment.

[0087] LOVO cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum and passaged every 2-3 days. The cell culture incubator setting conditions were: CO 2 Concentration: 5%, Temperature: 37 o C, Cells in the logarithmic growth phase were used for the experiment.

[0088] NCIH-1385 cells, PANC-1 cells, AsPc-1 cells, LOVO cells (8 × 10 3 After 24 hours of incubation, the substance to be measured was added and incubated for 48 hours. After the cell supernatant was removed, 100 μL of MTT aqueous solution with a final concentration of 0.5 mg / mL was added to each well, and the culture plate was placed in an incubator to continue incubating for 4 hours. After the cell supernatant was removed, 100 μL of DMSO was added, and the culture plate was shaken and the absorbance (OD) at 490 nm was measured with an enzyme marker. Cell activity % = OD value of the drug group / OD value of the blank group * 100%.

[0089] 2.2 Measurement results

[0090] Table 3 IC values ​​of preferred compounds against tumor cell proliferation 50

[0091]

[0092] Note: CDDP is the positive control drug cisplatin.

[0093] The novel quinoline compounds designed and synthesized by the present invention have certain growth inhibition effects on tumor cells. 50 The values ​​were basically around 10~30μM. Among them, C12 showed good anti-proliferative activity in 4 types of tumor cells, laying the foundation for the development of anti-cancer drugs.

Claims

1. A compound represented by general formula (I) or a pharmaceutically acceptable salt thereof, Formula (I) in, R is: 、 、 、 、 、 、 、 。 2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The R is: or .

3. A pharmaceutical composition comprising the compound according to any one of claims 1 to 2 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

4. The compound according to any one of claims 1 to 2 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 3 in the preparation of a drug for inhibiting KRAS in cells G12D Active pharmaceutical application.

5. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 2 or the pharmaceutical composition according to claim 3 in the preparation of KRAS G12D Application in drugs for related cancers.

6. The use according to claim 5, characterized in that: The KRAS G12D The relevant cancer is selected from: angiosarcoma, rhabdomyosarcoma, myxoma, rhabdomyoma, fibroma, teratoma, lung cancer, bronchogenic carcinoma, bronchial adenoma, esophageal cancer, gastric cancer, pancreatic cancer, colorectal cancer, small intestine cancer, large intestine cancer, rectal cancer, kidney cancer, leukemia, bladder and urethra cancer, prostate cancer, testicular cancer, liver cancer, gallbladder cancer, bone cancer, malignant fibrous histiocytoma, chondrosarcoma, malignant lymphoma, malignant giant cell tumor chordoma, osteochondroma, benign chondroma, chondroblastoma, chondromyxoid fibroma, glioma, germ cell tumor, glioblastoma multiforme, oligodendroglioma, neurothecoma, retinoblastoma, congenital tumors, spinal neurofibroma, endometrial cancer, malignant teratoma, vulvar squamous cell carcinoma, fallopian tube cancer, malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi sarcoma, lipoma, hemangioma.

7. The use according to claim 6, characterized in that: The cancer is lung cancer, colorectal cancer or pancreatic cancer.

8. The use according to claim 7, characterized in that: The lung cancer is non-small cell lung cancer or small cell lung cancer.

9. The use according to claim 7, characterized in that: The cancer is pancreatic cancer.

10. The use according to claim 7, characterized in that: The cancer is colorectal cancer.

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