Pan-KRAS inhibitors and their medical uses

Abstract

The present invention relates to the field of medicinal chemistry, to pan-KRAS inhibitors and their medical uses, and in particular to a class of KRAS-targeted and mutant KRAS G12C 、KRAS G12D The results of affinity pharmacological tests on KRAS and its mutants show that the compounds of the present invention have good affinity for KRAS wild type and its KRAS G12C Mutant, KRAS G12D The mutants showed better affinity than the lead compound for KRAS wild-type and its KRAS G12C Mutant, KRAS G12D Mutated targets have potential inhibitory effects.

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, to pan-KRAS inhibitors and their medical uses, and in particular to a class of KRAS-targeted and mutant KRAS G12C 、KRAS G12D Compounds and their medical uses. 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 switch from the inactive GDP-bound conformation to the active GTP-bound state, so that the signal is normally transmitted to the downstream cells. In tumor cells, RAS protein loses its inherent GTPase activity due to point mutations, thereby maintaining the GTP active state, causing abnormal activation of various downstream signals. There are three main types of RAS mutations: HRAS, KARS and NRAS. Among them, KRAS is the most common mutation in the RAS family, present in 22% of human tumors, especially in lung cancer, colorectal cancer and pancreatic cancer. KRAS regulates its activity through the exchange mechanism of GTP and GDP. In the activated state (KRAS-GTP), it binds to downstream effector molecules to initiate signal transduction. In the inactive state (KRAS-GDP), it is regulated by GTPase activating proteins (GAPs). The activation of KRAS involves the MAPK / ERK pathway and the PI3K / AKT pathway, which play an important role in cell growth, differentiation and survival. KRAS mutations cover multiple different sites, among which G12C, G12D and G12R mutations are the most common. KRAS and its mutants are potential drug targets. But on the one hand, it lacks a small molecule binder pocket, and on the other hand, it has a high affinity for GTP and GDP, so the development of drugs targeting KRAS and its mutants is a challenging task. In recent years, it has been discovered that there is a binding pocket P110 on the surface of KRAS. KRAS inhibitors can directly target the P110 pocket of the KRAS protein, preventing it from binding to GTP and maintaining the state of binding to GDP. The discovery of this site may break the long-term "undruggability" of KRAS.

[0003] Currently, research on KRAS mutations mainly focuses on the impact of different mutation types on disease prognosis and targeted therapies for these mutations. G12C and KRAS G12DThe development of mutation inhibitors aims to improve the survival and quality of life of patients. In addition, the application of immunotherapy in patients with KRAS mutations is also a research area that has attracted much attention, especially in patients with non-small cell lung cancer. There are many antagonists targeting KRAS developed at home and abroad, but their low selectivity or poor biological activity limits their further in-depth research. So far, there are only two antagonists targeting KRAS. G12C Drugs are on the market, but without KRAS and its KRAS G12D Inhibitors are on the market, so the discovery of novel and highly effective KRAS inhibitors is of great research significance.

[0004] The compound KAL-21404358 can specifically bind to the binding pocket P110 on the surface of KRAS, and after binding, it can disrupt the interaction between KRAS−B-Raf and destroy the KRAS−B-Raf as well as the Akt and Erk signaling pathways. This discovery and the development of related inhibitors will further break the cycle of 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:

[0005]

[0006] As the incidence of cancer is increasing year by year, its pathological mechanism is very complex, therefore, it is urgent to find KRAS inhibitors with diverse structures to alleviate cancer. The present invention uses KAL-21404358 as a lead compound and discovers a new type of substituted quinoline compound, which has good selectivity and anti-tumor cell proliferation activity against KRAS and its mutants. Summary of the invention

[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 shown in the general formula (I):

[0009]

[0010] Formula (I)

[0011] R is:

[0012] , .

[0013] Preferably, the 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 pharmaceutical composition comprising the compound described in the first aspect or a pharmaceutically acceptable salt, isomer or solvate thereof, and a pharmaceutically acceptable carrier.

[0018] 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.

[0019] "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.

[0020] The pharmaceutical composition is in the form of injection, capsule, tablet, pill, powder or granule.

[0021] 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.

[0022] 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.

[0023] 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.

[0024] 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.

[0025] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0026] 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.

[0027] 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.

[0028] 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.

[0029] The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds (such as anti-tumor drugs).

[0030] The treatment method of the present invention can be used alone or in combination with other treatment methods or therapeutic drugs.

[0031] 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.

[0032] The third aspect of the technical solution of the present invention is to provide use of the compound described in the first aspect and the pharmaceutical composition described in the second aspect in the preparation of a drug for treating and / or preventing diseases mediated by KRAS and its mutations.

[0033] Alternatively, a method for treating and / or preventing KRAS is provided, comprising the compound of the first aspect and the pharmaceutical composition of the second aspect. G12C Application of drugs in mutation-mediated diseases.

[0034] Alternatively, a method for treating and / or preventing KRAS is provided comprising the compound of the first aspect and the pharmaceutical composition of the second aspect. G12D Application of drugs in mutation-mediated diseases.

[0035] Diseases mediated by KRAS and its mutations include cancer and inflammatory diseases.

[0036] The cancer includes colon cancer, rectal cancer, lung cancer, and melanoma. The lung cancer is non-small cell lung cancer and small cell lung cancer.

[0037] Since the compounds of the present invention have excellent anti-cancer activity, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to treat, prevent and alleviate cancer-related diseases.

[0038] Compared with the prior art, the present invention has the following main advantages:

[0039] (1) The affinity of the compounds of the present invention to KRAS and its mutants is shown in Table 1. The synthetic WY2-036, WY2-038, WY2-041, WY2-044, WY2-050, WY2-069, and WY2-073 have a strong affinity to wild-type KRAS and its mutants. G12C Mutant, KRAS G12D The affinity of the mutants was better than that of the lead compound KAL-21404358, especially compounds WY2-038, WY2-041 and WY2-069 had the best affinity.

[0040] (2) The compounds WY2-038, WY2-069 and the lead compound KAL-21404358 described in the present invention have good inhibitory effects on human lung cancer (A549), human melanoma (A375) and human colon cancer (LOVO) cells. Among them, WY2-069 and the lead compound KAL-21404358 have better inhibitory effects on the corresponding cancer cells than cisplatin, and WY2-069 has the best inhibitory effect.

[0041] (3) The IC of the compounds on tumor cells was tested 50 The experimental results showed that compound WY2-069 has excellent anti-tumor activity, which is better than cisplatin, and has value in the development of KRAS-targeted anti-tumor drugs. DETAILED DESCRIPTION

[0042] 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.

[0043] 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.

[0044] 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.

[0045] 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.

[0046] Example 1

[0047] Synthesis of 4-(Benzyloxy)quinolin-2-yl)methanol (WY2-036):

[0048]

[0049] Methyl 4-oxo-1,4-dihydroquinoline-2-carboxylate (3) (4.9 mmol) was dissolved in MeOH (50 mL) and heated at 0. o NaBH4 (24.6 mmol) was added at 25 o C for 4 h. The organic phase was quenched with ice-saturated aqueous ammonium chloride solution, 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 2-(hydroxymethyl)quinolin-4(1H)-one (7). Compound 7 (1 mmol) was dissolved in acetonitrile (2 mL), anhydrous potassium carbonate was added, and the halogenated hydrocarbon reagent benzyl bromide was slowly added under nitrogen protection, and the reaction was refluxed for 12 h. The reaction solution was quenched with water, 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 compound WY2-36. The product is a white solid, with a yield of 30%. 1 HNMR (400 MHz, DMSO- d 6) δ 8.15 (dd,J = 8.3, 1.4 Hz, 1H), 7.90 (d, J = 8.4 Hz,1H), 7.71 (ddd, J = 8.4, 6.8, 1.5 Hz, 1H), 7.58 (d, J = 7.2 Hz, 2H), 7.55 – 7.36(m, 4H), 7.27 (s, 1H), 5.58 (s, 1H), 5.40 (s, 2H), 4.70 (s, 2H). IR (KBr, cm -1 ): 3093, 3061, 3034, 2924, 2853, 1620, 1595, 1568, 1557, 1508, 1487, 1470, 1464, 1454, 1427, 1391, 1323, 1 234, 1177, 1153, 1136, 1103, 1078, 1032, 1001, 984, 957, 910, 872, 851, 827, 781, 758, 745, 719, 696, 675. HRMS (ESI): m / z Calculate for C 17 H 16 NO2 + (M+H) + :266.1181, found: 266.1180. mp 123.5 o C.

[0050] The preparation methods of the compounds in Examples 2-7 are the same as those in Example 1, except that different halogenated hydrocarbon reagents are used. The specific halogenated hydrocarbon used in each example is described in the corresponding example.

[0051] Example 2

[0052] Synthesis of 4-((2-(hydroxymethyl)quinolin-4-yl)oxy)methyl)benzonitrile (WY2-038):

[0053]

[0054] The halogenated hydrocarbon used is 4-cyanobenzyl bromide. The product is a white solid, with a yield of 30%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.20 (dd, J = 8.3, 1.4 Hz, 1H), 7.91 (t, J= 7.9 Hz, 3H), 7.81 – 7.69(m, 3H), 7.54 (ddd, J = 8.2, 6.9, 1.2 Hz, 1H), 7.23 (s, 1H), 5.57 (t, J = 5.8 Hz,1H), 5.52 (s, 2H), 4.68 (d, J = 5.2 Hz, 2H). IR (KBr, cm -1 ): 3072, 2928, 2847, 2230, 1595, 1570, 1443, 1383, 1159, 1123, 1115, 1067, 972, 870, 856, 817, 754, 646, 546.HRMS (ESI): m / z Calculate for C 18 H 15 N2O2 + (M+H) + : 291.1134, found: 291.1132. mp187.7 o C.

[0055] Example 3

[0056] Synthesis of (4-((4-bromophenyl)oxy)quinolin-2-yl)methanol (WY2-041):

[0057]

[0058] The halogenated hydrocarbon used is p-bromobenzyl bromide. The product is a white solid, with a yield of 67%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.15 (dd, J = 8.3, 1.4 Hz, 1H), 7.89 (d, J = 8.4 Hz, 1H), 7.81 – 7.68 (m,1H), 7.68 – 7.57 (m, 2H), 7.60 – 7.42 (m, 3H), 7.23 (s, 1H), 5.55 (t, J = 5.8Hz, 1H), 5.39 (s, 2H), 4.67 (d, J = 5.4 Hz, 2H). IR (KBr, cm -1): 2922, 2851, 1620, 1572, 1595, 1508, 1489, 1427, 1406, 1379, 1354, 1302 , 1173, 1157, 1138, 1109, 1070, 1013, 986, 872, 847, 804, 764, 545, 473. HRMS (ESI): m / z Calculate for C 17 H 15 BrNO2 + (M+H) + :344.0286, found: 344.0281. mp 104.5 o C.

[0059] Example 4

[0060] Synthesis of (4-(4-bromobutoxy)quinolin-2-yl)methanol (WY2-044):

[0061]

[0062] The halogenated hydrocarbon used is 1,4-dibromobutane. The product is a white solid, yield: 67%, 1 H NMR (400 MHz, DMSO- d 6) δ 8.12 (dd, J = 8.4, 1.4 Hz, 1H), 7.89 (d, J = 8.4 Hz, 1H), 7.71 (ddd, J =8.4, 6.8, 1.5 Hz, 1H), 7.52 (ddd, J = 8.2, 6.8, 1.2 Hz, 1H), 7.13 (s, 1H), 5.58(s, 1H), 4.68 (s, 2H), 4.30 (t, J = 5.9 Hz, 2H), 3.66 (t, J = 6.3 Hz, 2H), 2.30 –1.82 (m, 4H). IR (KBr, cm -1): 3445, 3227, 3055, 2957, 2920, 2851, 2567, 2006,1923, 1879, 1730, 1715, 1643, 1597, 1495, 1472, 1433, 1412, 1383, 1366, 1325,1290, 1246, 1186, 1161, 1121, 1082, 968, 947, 889, 862, 826, 767, 617, 606.HRMS (ESI): m / z Calculate for C 14 H 17 BrNO2 + (M+H) + : 310.0443, found: 310.0445. mp124.3 o C.

[0063] Example 5

[0064] Synthesis of (4-(allyloxy)quinolin-2-yl)methanol (WY2-050):

[0065]

[0066] The halogenated hydrocarbon used is 3-bromopropylene. The product is a white solid, yield: 67%, 1 H NMR (600 MHz, DMSO- d 6) δ 8.15 (dd, J = 8.4, 1.4 Hz, 1H), 7.94 – 7.83 (m, 1H), 7.71 (ddd, J = 8.4,6.8, 1.5 Hz, 1H), 7.51 (ddd, J = 8.1, 6.8, 1.2 Hz, 1H), 7.14 (s, 1H), 6.27 –6.11 (m, 1H), 5.74 (s, 1H), 5.56 – 5.50 (m, 1H), 5.36 (dq, J = 10.6, 1.5 Hz,1H), 4.85 (dt, J = 5.2, 1.6 Hz, 2H), 4.68 (s, 2H). IR (KBr, cm -1): 2988, 2957, 2943, 2916, 2872, 1597, 15570, 1558, 1541, 1503, 1472, 1464, 1456, 1422, 1408, 1395, 1379, 1362, 1350, 13 41, 1323, 1300, 1263, 1234, 1179, 1155, 1138, 1109, 1080, 1032, 1020, 988, 949, 924, 895, 864, 845, 827, 758, 735, 598. HRMS (ESI): m / z Calculate for C 13 H 14 NO2 + (M+H) + :216.1025, found: 216.1021. mp 113.5 o C.

[0067] Example 6

[0068] Synthesis of (4-((4-(trifluoromethyl)benzyl)oxy)quinolin-2-yl)methanol (WY2-069):

[0069]

[0070] The halogenated hydrocarbon used is 4-trifluoromethylbenzyl bromide. The product is a white solid, yield: 67%, 1 H NMR (600 MHz, DMSO- d 6) δ 8.25 – 8.17 (m, 1H), 8.01 – 7.64 (m, 6H), 7.58 – 7.49 (m, 1H), 7.25 (s, 1H), 5.57 (s, 1H), 5.53 (s, 2H), 4.67 (d, J = 5.7 Hz, 2H). IR (KBr,cm -1 ): 2924, 2853, 1620, 1597, 1573, 1510, 1456, 1420, 1387, 1358, 1325, 1236, 1163, 1109, 1067, 1018, 988, 972, 841, 826, 766, 721, 592, 577. HRMS (ESI): m / z Calculate for C 18 H 15 F3NO2 + (M+H) +: 334.1055, found: 334.1054. mp 72.8 o C.

[0071] Example 7

[0072] Synthesis of (4-((4-methoxybenzyl)oxy)quinolin-2-yl)methanol (WY2-073):

[0073]

[0074] The halogenated hydrocarbon used is 4-methoxybenzyl bromide. The product is a white solid, yield: 67%, 1 H NMR (400 MHz, DMSO- d 6) δ 8.13 (d, J = 8.2 Hz, 1H), 7.95 (d, J = 8.5 Hz, 1H), 7.74 (dt, J = 21.6,7.7 Hz, 2H), 7.51 (d, J = 8.3 Hz, 2H), 7.32 (s, 1H), 7.01 (d, J = 8.4 Hz, 2H), 5.36 (s, 2H), 4.72 (s, 2H), 3.97 (s, 1H), 3.78 (s, 3H). IR (KBr, cm -1 ): 2955,2924, 2851, 2398, 2378, 2251, 1738, 1726, 1657, 1639, 1618, 1610, 1596, 1572,1562, 1553, 1528, 1512, 1501, 1493, 1476, 1468, 1450, 1441, 1369, 1342, 1321,1302, 1254, 1236, 1177, 1157, 1138, 1107, 1080, 1032, 980, 935, 881, 858,826, 806, 768, 615. HRMS (ESI): m / z Calculate for C 18 H 18 NO3 + (M+H) + : 296.1287, found:296.1283. mp 105.4 o C.

[0075] Some pharmacological tests and results of the compounds of the present invention are as follows:

[0076] Experimental Example 1: Effects of the compounds of the present invention on KRAS wild-type, KRAS G12C Mutant, KRAS G12D Mutant affinity testing.

[0077] 1.1 Experimental Principle:

[0078] Surface Plasmon Resonance (SPR) principle: A beam of P-polarized light is incident on the end face of a prism within a certain angle range, and a surface plasmon wave will be generated at the interface between the prism and the metal film (Au or Ag). When the propagation constant of the incident light wave matches the propagation constant of the surface plasmon wave, it causes the free electrons in the metal film to resonate, which is called surface plasmon resonance. During analysis, a layer of biomolecule recognition film is first fixed on the surface of the chip, and then the sample to be tested flows over the surface of the chip. If there are molecules in the sample that can interact with the biomolecule recognition film on the surface of the chip, it will cause the refractive index of the gold film surface to change, and ultimately lead to a change in the SPR angle. By monitoring the change in the SPR angle, the concentration, affinity, kinetic constant and specificity of the analyte can be obtained. Here we use molecular interaction technology to detect the synthesized compounds and KRAS protein to obtain the binding ability of the compounds with KRAS protein.

[0079] 1.2 Experimental method: Weigh an appropriate amount of the compound to be tested 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 compound to be tested as a blank control.

[0080] The Biacore T200 surface plasmon resonance spectrometer was used to analyze the interactions between the compounds and KRAS wild-type and KRAS G12C Mutant, KRAS G12D Mutant affinity test, measuring the affinity of the compound to the protein K D value.

[0081] 1.3 Experimental Results

[0082] Table 1 Affinity of compounds for KRAS

[0083]

[0084] Note: KAL-21404358 is the lead compound.

[0085] The affinity of the synthesized compounds to KRAS and its mutants is shown in Table 1. The synthetic WY2-036, WY2-038, WY2-041, WY2-044, WY2-050, WY2-069, and WY2-073 have a strong affinity to KRAS wild type and its KRAS G12C Mutant, KRAS G12D The affinity of the mutants is better than that of the lead compound KAL-21404358. In particular, compounds WY2-038, WY2-041, and WY2-069 have the best affinity and are the experimental basis for the development of new drugs.

[0086] Experimental Example 2: Inhibitory effect of target compound on tumor cell growth

[0087] 2.1 MTT experimental principle and method

[0088] MTT is 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide. It 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 marker, which can indirectly reflect the number of living cells. In short, the content of blue-purple crystalline formazan is positively correlated with the number of living cells.

[0089] 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 the following conditions: CO2 concentration: 5%, temperature: 37 o C, Cells in the logarithmic growth phase were used for the experiment.

[0090] 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: CO2 concentration: 5%, temperature: 37 o C, Cells in the logarithmic growth phase were used for the experiment.

[0091] LOVO 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: CO2 concentration: 5%, temperature: 37 o C, Cells in the logarithmic growth phase were used for the experiment.

[0092] A549 cells, A375 cells or LOVO cells (8 × 10 3Cells were inoculated into 96-well plates (100 μL / well / 100 μL). After incubation for 24 h, the compounds and cisplatin at the concentration to be tested were added and incubated for another 48 h. After the supernatant of the cultured cells was removed, 100 μL of MTT solution containing a final concentration of 0.5 mg / mL was added to each well. The culture plate was placed in an incubator and incubated for another 4 h. After the supernatant of the cells was removed, 100 μL of DMSO was added. After the culture plate was shaken, the absorbance (OD) at 490 nm was measured using an enzyme marker. Cell activity % = OD value of the drug group / OD value of the blank group * 100%.

[0093] 2.2 Experimental Results

[0094] Table 2-1 Inhibitory effect of target compounds (10 μM) on cancer cells

[0095]

[0096] Note: DDP is the positive control drug cisplatin. KAL-21404358 is the lead compound.

[0097] By observing the above experimental data, it was found that the representative compounds WY2-038, WY2-069 and the lead compound KAL-21404358 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 WY2-069 and the lead compound KAL-21404358 have better inhibitory effects on the corresponding cancer cells than cisplatin, among which WY2-069 has the best inhibitory effect, so the IC of the compound on tumor cells was further tested. 50 .

[0098] Table 2-2 IC of some compounds on tumor cells 50

[0099]

[0100] Note: DDP is the positive control drug cisplatin. KAL-21404358 is the lead compound.

[0101] From the experimental results, it can be seen that compound WY2-069 has excellent anti-tumor activity, which is better than cisplatin, and has value in the research and development of KRAS-targeted anti-tumor drugs.

[0102] 2.3 Results and Discussion

[0103] KRAS affinity experiments show that most of the synthesized new substituted quinoline compounds have ideal affinity for KRAS and its mutant isomers. Compounds with targeted affinity have potential inhibitory effects on targets and are the experimental basis for the test of anti-tumor cell proliferation activity of the preferred compounds. Among them, compounds WY2-038, WY2-041 and WY2-069 have the best affinity for KRAS and its mutants. The inhibitory effect on tumor cell proliferation shows that compound WY2-069 has a good inhibitory effect on human lung cancer (A549), human melanoma (A375) and human colon cancer (LOVO) cells, has excellent anti-tumor activity, and is superior to cisplatin, laying the foundation for the development of corresponding targeted anticancer drugs.

Claims

1. A compound represented by general formula (I) or a pharmaceutically acceptable salt thereof, Formula (I) 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. Use of 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 medicament for treating and / or preventing a disease mediated by KRAS and its mutations, wherein the disease mediated by KRAS and its mutations is cancer, and the cancer is colon cancer, lung cancer, or melanoma.

5. 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 G12C Application of drugs in diseases mediated by mutations, such as KRAS G12C The mutation-mediated disease is cancer, including colon cancer, lung cancer, and melanoma.

6. 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, such as KRAS G12D The mutation-mediated disease is cancer, including colon cancer, lung cancer, and melanoma.

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