Substituted quinolinone compounds targeting KRASG12D and their applications

Abstract

The present invention relates to the field of medicinal chemistry, and in particular to a class of drugs targeting KRAS G12D Substituted quinolinone compounds and their applications. Pharmacological experiments have shown that the compounds synthesized by the present invention have an effect on KRAS G12D The affinity of the mutants was better than that of the lead compound KAL-21404358. Compounds WY1-014 and WY1-062 had ideal inhibitory effects on human lung cancer (A549), human melanoma (A375) and human colon cancer (LOVO) cells. Compound WY1-062 had better antitumor activity, which was better than cisplatin.

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and in particular to a class of drugs targeting KRAS G12D Substituted quinolinone compounds and applications thereof. Background Art

[0002] RAS protein belongs to the small GTPase family and is closely related to the differentiation, proliferation and survival of downstream cells. RAS protein can convert from an inactive GDP-bound conformation to an active GTP-bound state, so that the signal is normally transmitted to downstream cells. In tumor cells, RAS protein loses its inherent GTPase activity due to point mutations, thereby maintaining the GTP active state, causing various downstream signals to be abnormally activated. There are three main RAS subtypes: HRAS, KRAS and NRAS. Among them, KRAS is the most common mutation subtype, present in 22% of human tumors, and is a potential drug target. However, because it lacks a small molecule binder pocket and has a high affinity for GTP and GDP, the development of targeted drugs for KRAS is a challenging task. Currently, there are many drugs targeting KRAS. G12C The research on inhibitors of RAS has made great progress. These achievements have not only transformed RAS protein from an undruggable to a druggable target, but also promoted the study of other subtypes such as KRAS. G12D Research on inhibitors of KRAS G12D As the most common mutation in KRAS, accounting for about 40% of KRAS mutations, it is also challenging. In recent years, it has been found that there is a binding pocket P110 on the surface of KRAS. KRAS inhibitors can directly target the P110 pocket of KRAS protein, preventing it from binding to GTP and maintaining the state of binding to GDP. The discovery of this site may break the KRAS G12D The phenomenon of “undruggability”.

[0003] The compound KAL-21404358 can specifically bind to the binding pocket P110 on the surface of KRAS, and after binding, it can affect the interaction between KRAS and B-Raf and disrupt the KRAS-B-Raf, Akt and Erk signaling pathways. This discovery and the development of related inhibitors will further break the KRAS, especially the KRAS mutant KRAS G12D On this basis, the P110 site may be an allosteric regulatory site for KRAS, providing a theoretical basis for the development of targeted drugs. The structure of the lead compound KAL-21404358 in this project is as follows:

[0004]

[0005] In this study, KAL-21404358 was used as the lead compound to synthesize a series of novel substituted quinolinone compounds, which have the potential to inhibit KRAS G12D The mutant has better affinity and anti-tumor cell proliferation activity. Summary of the invention

[0006] The technical problem solved by the present invention is to provide a class of targeted KRAS and its mutant KRAS G12C 、KRAS G12D Substituted quinolinone compounds, preparation methods and pharmaceutical compositions thereof, and use thereof in the preparation of drugs for preventing and / or treating diseases associated with abnormal function of KRAS and its mutants.

[0007] In order to solve the technical problem of the present invention, the present invention provides the following technical solutions:

[0008] The first aspect of the technical solution of the present invention is to provide a compound as represented by general formula (I) or a pharmaceutically acceptable salt, isomer or solvate thereof.

[0009]

[0010] Formula (I)

[0011] Where R is:

[0012] , .

[0013] Preferably, said R is: .

[0014] As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by a compound of the present invention and an acid or base that is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by a compound of the present invention and an acid. Suitable acids for forming salts include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, and naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid.

[0015] Another preferred salt is a salt of the compound of the present invention and a base, such as an alkali metal salt (e.g., sodium salt or potassium salt), an alkaline earth metal salt (e.g., magnesium salt or calcium salt), an ammonium salt (e.g., lower alkanolammonium salt and other pharmaceutically acceptable amine salts), for example, methylamine salt, ethylamine salt, propylamine salt, dimethylamine salt, trimethylamine salt, diethylamine salt, triethylamine salt, tert-butylamine salt, ethylenediamine salt, hydroxyethylamine salt, dihydroxyethylamine salt, trihydroxyethylamine salt, and amine salts formed from morpholine, piperazine, and lysine, respectively.

[0016] The term "solvate" refers to a complex in which the compound of the present invention is coordinated with solvent molecules to form a specific ratio.

[0017] The second aspect of the technical solution of the present invention is to provide a method for preparing the compound described in the first aspect. The compound of general formula (I) of the present invention can be prepared by the following method:

[0018]

[0019] Where R is:

[0020] , .

[0021] Aniline (1) and dimethyl acetylenedicarboxylate were used as raw materials, dissolved in methanol, and reacted at room temperature in the dark for 24 h to obtain compound (2); compound (2) and PPA were heated under reflux for 3 h to obtain compound (3); compound (3) was dissolved in NMP, reacted with NaH for 1 h, and then SEMCl was added to react for 3 h to obtain compound (4); compound (4) was added with DBAL-H at -60 °C and reacted for 6 h to obtain intermediate compound (5); intermediate compound (5) was subjected to reductive amination with different amines to obtain WY1 type target compounds.

[0022] The third aspect of the technical solution of the present invention is to provide a pharmaceutical composition comprising the compound described in the first aspect or a pharmaceutically acceptable salt, isomer or solvate thereof, and a pharmaceutically acceptable carrier.

[0023] The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 10-1000 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

[0024] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers and wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0025] The pharmaceutical composition is in the form of injection, capsule, tablet, pill, powder or granule.

[0026] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administrations include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0027] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0028] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0029] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0030] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0031] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0032] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0033] Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0034] The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds (such as anti-tumor drugs).

[0035] The treatment method of the present invention can be used alone or in combination with other treatment methods or therapeutic drugs.

[0036] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 50 to 1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

[0037] The fourth aspect of the technical solution of the present invention is to provide a method for preparing a drug for treating and / or preventing KRAS comprising the compound of the first aspect and the pharmaceutical composition of the third aspect. G12D Application of drugs in mutation-mediated diseases.

[0038] Among them, the KRAS G12D Diseases mediated by mutations include cancers and inflammatory diseases. The cancers include colon cancer, rectal cancer, lung cancer, breast cancer, liver cancer, stomach cancer, lung adenocarcinoma, esophageal cancer, skin cancer, melanoma, etc. Preferably, the cancer is colon cancer, rectal cancer, lung cancer, melanoma. The lung cancer is non-small cell lung cancer or small cell lung cancer.

[0039] Compared with the prior art, the present invention has the following main advantages:

[0040] KRAS G12D Mutant affinity experiments showed that the compounds synthesized by the present invention have a strong affinity for KRAS G12D The affinity of the mutants is better than that of the lead compound KAL-21404358. The results of tumor cell inhibition showed that compounds WY1-014 and WY1-062 have ideal inhibitory effects on human lung cancer (A549), human melanoma (A375) and human colon cancer (LOVO) cells. Compound WY1-062 has better anti-tumor activity, which is better than cisplatin, laying the foundation for the development of corresponding anti-cancer drugs. DETAILED DESCRIPTION

[0041] The present invention's various embodiments specifically describe the preparation methods of the compounds of the present invention, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combinations can be easily carried out by those skilled in the art to which the present invention belongs.

[0042] Unless otherwise specified, the raw materials and reagents used in the process for preparing the compounds of the present invention and the reagents used in the pharmacological experiments of the present invention can be purchased through commercial channels.

[0043] The experimental methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are calculated by weight.

[0044] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0045] Example 1

[0046] Synthesis of (S)-2-((2-(Hydroxymethyl)pyrrolidin-1-yl)methyl)quinolin-4(1H)-one (WY1-XN019)

[0047]

[0048] Aniline (1) (469.0 mmol) was dissolved in methanol (50 mL), and dimethyl acetylene dicarboxylate (704 mmol) was added. The reaction mixture was allowed to react for 24 h at room temperature. The solvent was evaporated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain a yellow oily compound, 2-(phenylamino) maleic acid dimethyl ester (2). (2) (2.4 mmol) was added to PPA (100 g) and the mixture was stirred at 25 °C for 24 h. o C, react for 3 h. The organic phase was washed with cold saturated sodium bicarbonate solution, ice water, suction filtered, and some impurities were washed with ether. The filter cake was dried and the obtained crude product was separated and purified by silica gel column chromatography to obtain a white solid compound 4-oxo-1,4-dihydroquinoline-2-carboxylic acid methyl ester (3). 3 (9.8 mmol) was dissolved in NMP (5 mL) and heated at 0. o Sodium hydride (14.8 mmol) was added at 25 o C, react for 1 h, add SEMCl (2 mL), and o C for 3 h. The organic phase was quenched with ice water, extracted with ethyl acetate, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The crude product was separated and purified by silica gel column chromatography to obtain a white solid compound 4-((2-(trimethylsilyl)ethoxy)methoxy)quinoline-2-carboxylic acid methyl ester (4). (4) (9.8 mmol) was dissolved in dichloromethane (50 mL) and heated to -60 °C. o C, DIBAL-H (19.6 mmol) was added, -60 o C for 6 h. The organic phase was quenched with saturated ammonium chloride, extracted with dichloromethane, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The crude product was separated and purified by silica gel column chromatography to obtain a white solid compound 4-((2-(trimethylsilyl)ethoxy)methoxy)quinoline-2-carboxaldehyde (5). (5) (1 mmol) was dissolved in DCM (10 mL), and the amine reagent L-prolinol (1.2 mmol) was added dropwise, and a few drops of glacial acetic acid were added. Sodium triacetoxyborohydride was added under nitrogen protection, and the reaction was carried out at room temperature for 2 h. The reaction solution was quenched with saturated ammonium chloride solution, extracted with dichloromethane, and the organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the solvent was evaporated under reduced pressure. The crude product was separated and purified by silica gel column chromatography to obtain the target compound WY1-XN019. The product was a colorless oily liquid with a yield of 30%. 1 H NMR (600 MHz, DMSO- d 6 ) δ 11.32 (s, 1H), 8.05 (dt, J= 7.9, 1.1 Hz,1H), 7.64 (ddd, J = 13.5, 8.7, 7.2 Hz, 2H), 7.29 (ddd, J = 8.0, 6.5, 1.4 Hz, 1H), 6.08 (s, 1H), 4.62 (s, 1H), 3.43 (s, 2H), 3.07 (d, J = 118.9 Hz, 2H), 2.73 (s,1H), 2.50 (p, J = 1.9 Hz, 2H), 1.98 – 1.56 (m, 4H). IR (KBr, cm -1 ): 3416, 3063, 2959, 2874, 2818, 1634, 1609, 1518, 1474, 1445, 1383, 1354, 1317, 1250, 1134, 1090, 1061, 1032, 993, 870, 853, 793, 762, 675. HRMS (ESI): m / z Calculate for C 15 H 19 N 2 O 2 + (M+H) + :259.1447 found: 259.1438.

[0049] The preparation methods of the compounds in Examples 2-20 are the same as those in Example 1, except that different amine reagents are used for the synthesis. The specific amine reagents used in each example are described in the corresponding example:

[0050] Example 2

[0051] Synthesis of 2-((3-methoxyphenyl)amino)methyl)quinolin-4(1H)-one (WY1-XN026)

[0052]

[0053] The amine used is m-aminoanisole. The product is a white solid, yield: 67%, 1 H NMR (400 MHz, DMSO- d 6 ) δ 11.57 (s, 1H), 8.03 (dd, J = 8.1, 1.3 Hz, 1H), 7.65 – 7.58 (m, 2H), 7.28(ddd, J= 8.1, 6.3, 1.7 Hz, 1H), 7.04 – 6.94 (m, 1H), 6.31 (t, J = 6.2 Hz, 1H), 6.25 – 6.17 (m, 3H), 6.06 (s, 1H), 4.28 (d, J = 5.5 Hz, 2H), 3.64 (s, 3H). IR(KBr, cm -1 ): 3319, 3067, 2918, 2833, 1724, 1647, 1601, 1541, 1508, 1468, 1431,1350, 1312, 1298, 1242, 1213, 1157, 1134, 1121, 1040, 1009, 989, 918, 824,758, 733, 683, 669, 604. HRMS (ESI): m / z Calculate for C 17 H 17 N 2 O 2 + (M+H) + : 281.1290found: 281.1283. mp 187.9 o C.

[0054] Example 3

[0055] Synthesis of 2-(((2-methoxyphenyl)amino)methyl)quinolin-4(1H)-one (WY1-014)

[0056]

[0057] The amine reagent used is o-anisidine. The product is a white solid with a yield of 67%. 1 H NMR (400 MHz, DMSO- d 6 ) δ 11.57 (s, 1H), 8.02 (dd, J = 8.1, 1.4 Hz, 1H), 7.63 – 7.56 (m, 2H), 7.28 (ddd, J = 8.1, 6.5, 1.5 Hz, 1H), 6.84 (dd, J = 8.0, 1.3 Hz, 1H), 6.71 (td, J =7.7, 1.4 Hz, 1H), 6.58 (td,J = 7.7, 1.5 Hz, 1H), 6.47 (dd, J = 7.8, 1.5 Hz, 1H), 6.04 (s, 1H), 5.74 (d, J = 8.2 Hz, 1H), 4.33 (d, J = 4.0 Hz, 2H), 3.82 (s, 3H).IR (KBr, cm -1 ): 3433, 3063, 2924, 2853, 1641, 1597, 1549, 1470, 1506, 1445, 1418, 1354, 1321, 1302, 1250, 1223, 1177, 1136, 1128, 1051, 1028, 986, 847, 760, 727. HRMS (ESI): m / z Calculate for C 17 H 17 N 2 O 2 + (M+H) + : 281.1290. found: 281.1278. mp 188.3 o C.

[0058] Example 4

[0059] Synthesis of 2-(((2-(Hydroxymethyl)phenyl)amino)methyl)quinolin-4(1H)-one (WY1-016)

[0060]

[0061] The amine reagent used is o-aminobenzyl alcohol. The product is a white solid, yield: 30%, 1 H NMR (600 MHz, DMSO- d 6 ) δ (ppm): 1 H NMR (400 MHz, DMSO- d 6 ) δ 11.59 (s, 1H), 8.04 (dd, J = 8.2,1.4 Hz, 1H), 7.68 – 7.54 (m, 2H), 7.28 (ddd, J = 8.1, 6.7, 1.3 Hz, 1H), 7.14(dd, J= 7.5, 1.6 Hz, 1H), 7.03 (td, J = 7.8, 1.6 Hz, 1H), 6.60 (t, J = 7.3 Hz,1H), 6.50 (d, J = 8.1 Hz, 1H), 6.08 (s, 1H), 5.86 (t, J = 6.2 Hz, 1H), 5.22 (t, J =5.4 Hz, 1H), 4.55 (d, J = 5.3 Hz, 2H), 4.39 (d, J = 6.1 Hz, 2H). IR (KBr, cm -1 ):2922, 2850, 1636, 1609, 1564, 1512, 1445, 1303, 1350, 1327, 1265, 1236, 1138, 1015, 982, 953, 928, 858, 793, 752, 735, 671, 637, 617. HRMS (ESI): m / z Calculate for C 17 H 17 N 2 O 2 + (M+H) + :281.1290. found: 281.1278. mp 213.4 o C.

[0062] Example 5

[0063] Synthesis of 2-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)quinolin-4(1H)-one (WY1-017)

[0064]

[0065] The amine reagent used is N-hydroxyethylpiperazine. The product is a white solid with a yield of 67%. 1 H NMR (400 MHz, DMSO- d 6 ) δ 11.50 (s, 1H), 8.04 (d, J = 8.0 Hz, 1H), 7.63 (dt, J = 15.1, 8.1 Hz,2H), 7.28 (t, J= 7.4 Hz, 1H), 6.04 (s, 1H), 3.52 (d, J = 21.1 Hz, 4H), 2.13 –1.79 (m, 2H), 1.29 (d, J = 46.9 Hz, 8H). IR (KBr, cm -1 ): 3144, 3046, 2805, 2006, 1991, 1769, 1760, 1715, 1633, 1557, 1539, 1506, 1470, 1454, 1447, 1404, 1140, 1123, 1082, 986, 947, 797, 760. HRMS (ESI): m / z Calculate for C 16 H 22 N 3 O 2 + (M+H) + : 288.1712 found:288.1694. mp 203.3 o C.

[0066] Example 6

[0067] Synthesis of 2-((Bis(2-hydroxyethyl)amino)methyl)quinolin-4(1H)-one (WY1-021)

[0068]

[0069] The amine reagent used is diethanolamine. The product is a white solid, with a yield of 30%. 1 H NMR (400 MHz, DMSO- d 6 ) δ 11.19 (s, 1H), 8.05 (d, J = 8.0 Hz, 1H), 7.62 (d, J = 3.8 Hz, 2H), 7.29(ddd, J = 8.1, 4.9, 3.2 Hz, 1H), 6.04 (s, 1H), 4.62 (t, J = 5.4 Hz, 2H), 3.75 (s,2H), 3.51 (q, J = 5.6 Hz, 4H), 3.37 (s, 4H). IR (KBr, cm -1):3116, 3057, 2924,2852, 1716, 1699, 1625, 1597, 1558, 1489, 1471, 1361, 1319, 1246, 1138, 1074,840, 752, 567, 459.HRMS (ESI): m / z Calculate for C 14 H 19 N 2 O 3 + (M+H) + : 263.1396 found:263.1389. mp 215.4 o C.

[0070] Example 7

[0071] Synthesis of 2-((Benzyl(2-hydroxyethyl)amino)methyl)quinolin-4(1H)-one (WY1-022)

[0072]

[0073] The amine reagent used is N-phenylethanolamine. The product is a white solid, with a yield of 30%. 1 H NMR (400 MHz, DMSO- d 6 ) δ11.75 (s, 1H), 8.03 (dd, J = 8.1, 1.4 Hz, 1H), 7.62 (ddd, J = 8.4, 6.9,1.5 Hz, 1H), 7.53 (d, J = 8.3 Hz, 1H), 7.29 (t, J = 7.5 Hz, 1H), 7.15 (dd, J = 8.6,7.0 Hz, 2H), 6.71 (d, J = 8.2 Hz, 2H), 6.63 (t, J = 7.2 Hz, 1H), 5.91 (s, 1H), 5.19 (s, 1H), 4.60 (s, 2H), 3.70 (d, J = 5.7 Hz, 2H), 3.60 (t, J = 5.7 Hz, 2H).IR(KBr, cm -1): 3063, 2924, 2856, 1716, 1697, 1683, 1637, 1595, 1458, 1396, 1386, 1246, 1232, 1163, 1134, 1101, 1070, 1033, 993, 879, 848, 765, 746. HRMS (ESI): m / z Calculate for C 18 H 19 N 2 O 2 + (M+H) + : 295.1447 found: 295.1443. mp 216.3 o C.

[0074] Example 8

[0075] Synthesis of 2-((3-methoxybenzyl)amino)methyl)quinolin-4(1H)-one (WY1-023)

[0076]

[0077] The amine reagent used is 3-methoxybenzylamine. The product is a white solid, yield: 30%, 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.09 – 8.01 (m, 1H), 7.61 (dd, J = 5.0, 1.8 Hz, 2H), 7.34 – 7.18 (m,2H), 6.99 – 6.88 (m, 2H), 6.79 (dd, J = 8.2, 2.6 Hz, 1H), 6.09 (s, 1H), 3.78 –3.66 (m, 7H). IR (KBr, cm -1 ): 3319, 2918, 2849, 2791, 2773, 1734, 1718, 1647,1638, 1597, 1560, 1543, 1508, 1501, 1491, 1474, 1468, 1460, 1439, 1354, 1315,1283, 1261, 1244, 1152, 1134, 1040, 841, 777, 760, 693, 530. HRMS (ESI): m / z Calculate for C 18 H19 N 2 O 2 + (M+H) + : 295.1447 found: 295.1443. mp 188.6 o C.

[0078] Example 9

[0079] Synthesis of 2-((4-methoxybenzyl)amino)methyl)quinolin-4(1H)-one (WY1-024)

[0080]

[0081] The amine reagent used is 4-methoxybenzylamine. The product is a white solid, with a yield of 30%. 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.08 – 8.04 (m, 1H), 7.61 (dd, J = 5.9, 1.6 Hz, 2H), 7.28 (dd, J =8.3, 6.4 Hz, 3H), 6.91 – 6.85 (m, 2H), 6.09 (s, 1H), 3.72 (s, 3H), 3.67 (d, J =7.4 Hz, 4H). IR (KBr, cm -1 ): 3065, 2928, 2833, 1724, 1630, 1560, 1553, 1510,1470, 1443, 1354, 1317, 1302, 1246, 1177, 1134, 1109, 1032, 995, 816, 760,675. HRMS (ESI): m / z Calculate for C 18 H 19 N 2 O 2 + (M+H) + : 295.1447 found: 295.1443. mp188.9 o C.

[0082] Example 10

[0083] Synthesis of 2-((2-(2-hydroxyethyl)phenyl)amino)methyl)quinolin-4(1H)-one (WY1-026)

[0084]

[0085] The amine reagent used is 2-aminophenylethanol. The product is a white solid with a yield of 30%. 1 H NMR (400 MHz, DMSO- d 6 ) δ 11.54 (s, 1H), 8.02 (dd, J = 8.1, 1.5 Hz, 1H), 7.71 – 7.50 (m, 2H), 7.27 (ddd, J = 8.1, 6.8, 1.2 Hz, 1H), 7.07 – 6.90 (m, 2H), 6.55 (td, J = 7.4, 1.1Hz, 1H), 6.50 – 6.37 (m, 1H), 6.03 (d, J = 1.6 Hz, 1H), 5.88 (s, 1H), 4.79 (t, J = 5.0 Hz, 1H), 4.33 (d, J = 6.1 Hz, 2H), 3.67 (td, J = 6.8, 4.9 Hz, 2H), 2.75 (t, J = 6.9 Hz, 2H). IR (KBr, cm -1 ): 3074, 2988, 2988, 2928, 1716, 1697, 1682, 1645, 1606, 1557, 1518, 1458, 1439, 1244, 1058, 1012, 849, 841, 810, 762, 743. HRMS (ESI): m / z calcdfor C 18 H 19 N 2 O 2 + (M+H) + : 295.1447 found: 295.1434. mp 220.3 o C.

[0086] Embodiment 11

[0087] Synthesis of 2-((4-methylpiperidin-1-yl)methyl)quinolin-4(1H)-one (WY1-032)

[0088]

[0089] The amine reagent used is 4-methylpiperidinium. The product is a white solid with a yield of 30%. 1 H NMR (400 MHz, DMSO- d 6 ) δ 11.41 (s, 1H), 8.04 (dd, J = 8.1, 1.4 Hz, 1H), 7.69 – 7.58 (m, 2H),7.28 (t, J = 7.4 Hz, 1H), 6.03 (s, 1H), 2.88 – 2.79 (m, 2H), 2.04 (t, J = 11.5Hz, 2H), 1.63 – 1.55 (m, 2H), 1.37 – 1.19 (m, 5H), 0.89 (d, J = 6.4 Hz, 3H). IR(KBr, cm -1 ): 3119, 3065, 2951, 2918, 2870, 2814, 1628, 1603, 1558, 1528, 1504, 1472, 1443, 1368, 1339, 1317, 1281, 1250, 1153, 1139, 1114, 1082, 1028, 978, 878, 841, 822, 770, 704, 675, 563, 465. HRMS (ESI): m / z Calculate for C 16 H 21 N 2 O + (M+H) + : 257.1654 found:257.1648. mp 210.7 o C.

[0090] Example 12

[0091] Synthesis of 2-((diisobutylamino)methyl)quinolin-4(1H)-one (WY1-033)

[0092]

[0093] The amine reagent used is diisobutylamine. The product is a white solid, yield: 30%, 1 H NMR (600 MHz, DMSO- d 6 ) δ 11.23 (s, 1H), 8.05 (dd,J = 8.2, 1.5 Hz, 1H), 7.71 – 7.55 (m, 2H), 7.29(ddd, J = 8.1, 6.6, 1.4 Hz, 1H), 6.15 (s, 1H), 3.52 (s, 2H), 2.18 (d, J = 7.1 Hz,4H), 1.77 (dt, J = 13.4, 6.8 Hz, 2H), 0.87 (d, J = 6.6 Hz, 12H). IR (KBr, cm -1 ):3082, 3067, 2953, 2926, 2868, 2795, 1634, 1595, 1558, 1508, 1474, 1447, 1387, 1354, 1314, 126 1, 1148, 1200, 1165, 1136, 1096, 1078, 1026, 993, 951, 858, 841, 818, 756, 716, 667, 637, 590, 546. HRMS (ESI): m / z Calculate for C 18 H 27 N 2 O + (M+H) + : 287.2123 found: 287.2120.mp 171.2 o C.

[0094] Example 13

[0095] Synthesis of 2-((Dimethylamino)methyl)quinolin-4(1H)-one (WY1-061)

[0096]

[0097] The amine reagent used is dimethylamine. The product is a white solid, with a yield of 67%. 1 H NMR (600 MHz, DMSO- d 6 )δ 11.46 (s, 1H), 8.04 (dd, J = 8.1, 1.5 Hz, 1H), 7.80 – 7.64 (m, 1H), 7.61(ddd, J = 8.5, 6.9, 1.5 Hz, 1H), 7.28 (ddd, J= 8.0, 6.9, 1.1 Hz, 1H), 6.01 (s,1H), 3.43 (s, 2H), 2.25 (s, 6H). IR (KBr, cm -1 ): 3063, 2967, 2940, 2818, 2770, 1636, 1597, 1557, 1508, 1472, 1441, 1350, 1317, 1267, 1252, 1 238, 1167, 1144, 1134, 1094, 1042, 993, 982, 889, 870, 853, 833, 760, 750, 716, 673, 573, 476. HRMS (ESI): m / z Calculate for C 12 H 15 N 2 O + (M+H) + : 203.1184 found: 203.1183. mp 115.3 o C.

[0098] Embodiment 14

[0099] Synthesis of 4-((4-oxo-1,4-dihydroquinolin-2-yl)methyl)amino)methyl)benzonitrile (WY1-062)

[0100]

[0101] The amine reagent used is 4-cyanobenzylamine. The product is a white solid, yield: 67%, 1 H NMR (600 MHz, DMSO- d 6 ) δ 8.05 (d, J IR (KBr, cm) -1):3277, 3073, 2984, 2957, 2924, 2862, 2824, 2351, 2320, 2220, 1726, 1659, 1641,1632, 1549, 1512, 1503, 1495, 1485, 1443, 1414, 1356, 1331, 1240, 1165, 1138,1092, 1016, 999, 974, 849, 816, 797, 762, 706, 675, 654. HRMS (ESI): m / z calcdfor C 18 H 16 N 3 O + (M+H) + : 290.1293 found: 290.1295. mp 184.4 o C.

[0102] Embodiment 15

[0103] Synthesis of 2-((Cyclohexylamino)methyl)quinolin-4(1H)-one (WY1-063)

[0104]

[0105] The amine reagent used is cyclohexylamine. The product is a white solid, with a yield of 67%. 1 H NMR (600 MHz, DMSO- d 6 )δ 8.07 (d, J = 8.0 Hz, 1H), 7.71 – 7.60 (m, 2H), 7.35 (t, J = 7.5 Hz, 1H), 6.29(s, 1H), 4.13 (d, J = 9.9 Hz, 2H), 2.39 (q, J = 1.9 Hz, 1H), 2.05 (d, J = 10.6 Hz,1H), 1.79 – 1.72 (m, 2H), 1.62 – 1.57 (m, 1H), 1.34 – 1.12 (m, 7H). IR (KBr,cm -1): 3069, 2922, 2851, 2658, 1639, 1595, 1547, 1499, 1470, 1443, 1354, 1317, 1240, 1136, 1107, 1026, 995, 962, 955, 887, 835, 754, 675. HRMS (ESI): m / z Calculate for C 16 H 21 N 2 O + (M+H) + : 257.1654 found: 257.1653.

[0106] Example 16

[0107] Synthesis of 2-((3-bromophenyl)amino)methyl)quinolin-4(1H)-one (WY1-065)

[0108]

[0109] The amine reagent used is 3-bromobenzylamine. The product is a white solid, yield: 67%, 1 H NMR (600 MHz, DMSO- d 6 ) δ 9.73 (s, 1H), 8.10 (dt, J = 8.3, 1.9 Hz, 1H), 7.78 (d, J = 1.9 Hz, 1H),7.72 (tt, J = 7.2, 1.8 Hz, 1H), 7.64 (dq, J = 8.4, 3.5, 3.0 Hz, 2H), 7.53 (d, J =7.6 Hz, 1H), 7.41 (dt, J = 12.5, 7.8 Hz, 2H), 6.42 (d, J = 11.0 Hz, 1H), 4.35 –4.23 (m, 4H). IR (KBr, cm -1): 2968, 2804, 2750, 2631, 2394, 2349, 1670, 1612,1571, 1554, 1530, 1472, 1447, 1360, 1339, 1317, 1213, 1192, 1130, 1076, 1032,968, 900, 878, 856, 843, 833, 792, 783, 764, 723, 694, 679, 671, 646.

[0110] Embodiment 17

[0111] Synthesis of 2-((3-chlorobenzyl)amino)methyl)quinolin-4(1H)-one (WY1-066)

[0112]

[0113] The amine reagent used is 3-chlorobenzylamine. The product is a white solid, yield: 67%, 1 H NMR (600 MHz, DMSO- d 6 ) δ 8.12 – 7.96 (m, 1H), 7.61 (dd, J = 5.7, 1.6 Hz, 2H), 7.46 (d, J = 1.8 Hz,1H), 7.41 – 7.20 (m, 4H), 6.08 (s, 1H), 3.71 (d, J = 29.0 Hz, 4H). IR (KBr, cm -1 ): 3435, 3269, 3067, 2920, 2851, 2367, 2345, 1638, 1630, 1597, 1560, 1543,1508, 1501, 1491, 1474, 1460, 1439, 1431, 1354, 1317, 1248, 1200, 1165, 1136,1026, 997, 961, 843, 783, 760, 698, 679. HRMS (ESI): m / z Calculate for C 17 H 16 C1N 2 O + (M+H) + : 299.0951 found: 299.0948. mp 205.3 o C.

[0114] Embodiment 18

[0115] Synthesis of 2-((4-chlorobenzyl)amino)methyl)quinolin-4(1H)-one (WY1-067)

[0116]

[0117] The amine reagent used is p-chlorobenzylamine. The product is a white solid, yield: 67%, 1 H NMR (600 MHz, DMSO- d 6 ) δ 8.04 (d, J = 8.0 Hz, 1H), 7.61 (d, J = 4.0 Hz, 2H), 7.38 (d, J = 5.6 Hz, 4H),7.28 (dd, J = 8.3, 3.9 Hz, 1H), 6.08 (s, 1H), 3.69 (d, J = 22.5 Hz, 4H). IR (KBr,cm -1 ): 3260, 3061, 2924, 2855, 2826, 1634, 1593, 1537, 1493, 1470, 1441, 1406,1352, 1331, 1306, 1290, 1242, 1167, 1138, 1088, 1015, 997, 961,874, 843, 818,804, 758, 677, 657. HRMS (ESI): m / z Calculate for C 17 H 16 C1N 2 O + (M+H) + : 299.0951 found:299.0946. mp 205.5 o C.

[0118] Embodiment 19

[0119] Synthesis of 2-((2-fluorobenzyl)amino)methyl)quinolin-4(1H)-one (WY1-068)

[0120]

[0121] The amine reagent used is 2-fluorobenzylamine. The product is a white solid, yield: 67%,1 H NMR (600 MHz, DMSO- d 6 ) δ 8.05 (dd, J = 8.2, 2.6 Hz, 1H), 7.61 (d, J = 4.0 Hz, 2H), 7.51 (td, J = 7.6,1.8 Hz, 1H), 7.33 – 7.25 (m, 2H), 7.22 – 7.12 (m, 2H), 6.10 (d, J = 3.4 Hz,1H), 3.74 (d, J = 30.7 Hz, 4H). IR (KBr, cm -1 ): 3065, 2922, 2830, 1716, 1697, 1681, 1668, 1645, 1560, 1551, 1508, 1473, 1456, 1439, 1396, 1354, 1317, 1234, 1179, 1136, 849, 756. HRMS (ESI): m / z Calculate for C 17 H 16 FN 2 O + (M+H) + : 283.1247 found: 283.1247. mp207.3 o C.

[0122] Embodiment 20

[0123] Synthesis of 2-([1,4'-bipiperidinyl]-1'-methyl)quinolin-4(1H)-one (WY1-091)

[0124]

[0125] The amine reagent used is 4-piperidinylpiperidine. The product is a white solid with a yield of 67%. 1 H NMR (400 MHz, DMSO- d 6 ) δ 12.08 (s, 1H), 8.07 (t, J = 7.8 Hz, 1H), 7.94 (d, J = 8.5 Hz, 1H),7.70 (ddd, J= 16.9, 6.8, 3.6 Hz, 2H), 6.63 (s, 1H), 4.23 (d, J = 6.6 Hz, 1H), 3.97 (s, 2H), 2.05 – 0.78 (m, 17H), 0.08 – -0.09 (m, 1H). IR (KBr, cm -1 ):2961, 2930, 2874, 2857, 1730, 1601, 1564, 1470, 1385, 1287, 1123, 1074, 962,941, 839, 787, 743, 704, 638, 534. HRMS (ESI): m / z Calculate for C 20 H 28 N 3 O + (M+H) + :326.2232 found:326.2236. mp 206.8 o C.

[0126] The pharmacological tests and results of the compounds of the present invention are as follows:

[0127] Experimental Example 1: Effects of the compounds of the present invention on KRAS G12D Mutant affinity testing.

[0128] 1.1 Experimental method: Weigh an appropriate amount of the test compound and add DMSO to prepare a 25 μM solution. Pipette an appropriate amount of the solution into a 96-well plate and dilute it to 200, 100, 50, 25, 12.5, 6.25, and 3.125 μM in sequence. At the same time, set DMSO without the test compound as a blank control.

[0129] Compound KRAS was analyzed by Biacore T200 surface plasmon resonance G12D Mutant affinity test. The K of the compound's affinity to the protein was measured. D value.

[0130] 1.2 Experimental Results

[0131] Table 1 Affinity of target compounds to KRAS

[0132]

[0133] Note: KAL-21404358 is the lead compound.

[0134] K D The smaller the value, the more related to KRASG12D The smaller the concentration of compound required for mutant affinity, the more likely the compound will bind to KRAS. G12D As shown in Table 1, the compounds synthesized by the present invention have a stronger affinity for KRAS G12D The affinity of the mutants was better than that of the lead compound KAL-21404358.

[0135] Experimental Example 2: Inhibitory effect of target compound on tumor cell growth

[0136] 2.1MTT Experiment Principle and Method

[0137] MTT is 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide, which can determine the survival status of cells and is particularly suitable for detecting chemical substances with cytotoxic functions. Succinate dehydrogenase in living cells can make MTT form blue-purple crystalline formazan, and this crystal is precipitated in the protein. In dead cells, there is no such enzyme. This color reaction is mainly carried out on living cells. After the formazan in the cells is decomposed by dimethyl sulfoxide (DMSO), its absorbance is measured by an enzyme reader, which can indirectly reflect the number of living bacteria. In short, the content of blue-purple crystalline formazan is positively correlated with the number of living bacteria.

[0138] A549 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.

[0139] A375 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.

[0140] 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.

[0141] A549 cells, A375 cells or LOVO cells (8 × 10 3After 24 hours of incubation, the compound to be measured and cisplatin were added and incubated for 48 hours. After the supernatant of the cultured cells was removed, 100 μL of MTT solution with a final concentration of 0.5 mg / mL was added to each well, and the culture plate was placed in an incubator for another 4 hours. After the supernatant of the cells 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-labeled instrument. Cell activity % = OD value of the drug group / OD value of the blank group * 100%.

[0142] 2.2 Experimental Results

[0143] Table 2-1 Inhibitory effect of target compounds (10 μM) on tumor cells

[0144]

[0145] Note: DDP is the positive control drug cisplatin. KAL-21404358 is the lead compound.

[0146] By observing the above experimental data, it was found that the three compounds WY1-014, WY1-062 and KAL-21404358 in the series of target compounds all had good inhibitory effects on human lung cancer (A549), human melanoma (A375) and human colon cancer (LOVO) cells. And from the data, it can be seen that the inhibitory effects of WY1-014, WY1-062 and KAL-21404358 on the corresponding tumor cells are better than cisplatin. Then the IC of the preferred compounds on tumor cells was tested. 50 .

[0147] Table 2-2 IC of some compounds on tumor cells 50

[0148]

[0149] Note: DDP is the positive control drug cisplatin. KAL-21404358 is the lead compound.

[0150] It can be seen from the experimental results that compound WY1-062 has excellent anti-tumor activity, which is better than cisplatin.

[0151] 2.3 Results and Discussion

[0152] KRAS G12D Mutant affinity experiments showed that the compounds synthesized by the present invention have a strong affinity for KRAS G12DThe affinity of the mutants is better than that of the lead compound KAL-21404358. The results of tumor cell inhibition showed that compounds WY1-014 and WY1-062 have ideal inhibitory effects on human lung cancer (A549), human melanoma (A375) and human colon cancer (LOVO) cells. Compound WY1-062 has better anti-tumor activity, which is better than cisplatin, laying the foundation for the development of corresponding 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: .

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 for use in the preparation of a drug for treating and / or preventing KRAS G12D Application of drugs in diseases mediated by mutations, described by KRAS G12D The mutation-mediated disease is cancer, including colon cancer, lung cancer, and melanoma.

5. The use according to claim 4, characterized in that: The lung cancer is non-small cell lung cancer or small cell lung cancer.

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