KRAS protein inhibitors and their use in preparing drugs for treating cancer

Abstract

The present invention belongs to the field of medicinal chemistry, and particularly relates to KRAS protein inhibitors and their use in the preparation of drugs for treating cancer. The compound shown by Formula I or the stereoisomer, tautomer, N-oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound shown by Formula I provided by the present invention can effectively stabilize the G-quadruplex structure in the KRAS promoter region, thereby down-regulating the transcription level of KRAS mRNA and effectively inhibiting the expression of KRAS protein, and can be used as a KRAS inhibitor. At the same time, this compound can be used as a highly effective inhibitor against all KRAS mutation types and can inhibit the proliferation of tumor cells. The structure of the compound shown by Formula I is as follows, wherein R1, R2, R3, and R4 are each independently selected from optionally substituted C 1‑10 alkoxy, and at least one of R1, R2, R3, and R4 is a hydroxyl group, #imgabs0#

Description

Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and in particular, the present invention relates to KRAS protein inhibitors and uses thereof in preparing drugs for treating cancer. Background Art

[0002] Tumor is one of the diseases that seriously endangers human health worldwide. In recent years, with the development of disciplines such as chemical biology, molecular oncology, and molecular pharmacology, the research and development of new anti-tumor drugs has made great progress. However, facing the most serious threat to human life and health, solid tumors account for more than 90% of malignant tumors, there is still a lack of highly effective and specific drugs. On the one hand, this reflects the difficulty of anti-tumor drug research and development, and on the other hand, it also means that the pace of research on the discovery and application of new anti-tumor targets needs to be accelerated.

[0003] The discovery of Ras family genes is a key milestone in tumor research (Cancer Res. 2012, 72: 2457-2467). Ras genes are common driver genes in malignant tumors. Mutations in Ras genes can cause cells to permanently "turn on" switches, leading to uncontrolled malignant proliferation and division of cells, resulting in carcinogenesis (Nat. Rev. Cancer, 2018: 767-777). To date, approximately 30% of human cancers are associated with Ras gene mutations, and most occur in pancreatic cancer, lung cancer, and colon cancer. The Ras gene family includes three genes: NRAS, HRAS, and KRAS, and Ras mutations mainly occur in the KRAS subtype (Nat. Rev. Drug Discov. 2014, 13: 828-851). KRAS is a key mediator of signal cascades that promote cell growth and proliferation, accounting for 85% of all Ras-related tumors. KRAS gene mutations are mainly concentrated at codons 12, 13 and 61, of which codon 12 mutations account for more than 80%, including G12A, G12C, G12D, G12R, G12S and G12V mutations. KRAS G12C mutations account for about 44% of all KRAS mutations and are most common in non-small cell lung cancer (Science, 2017, 355(6330): 1158-1163).

[0004] The protein encoded by KRAS in the cell switches between inactive and activated states. When KRAS binds to guanosine diphosphate (GDP), it is inactive. When it binds to guanosine triphosphate (GTP), it is inactive and activates downstream signaling pathways, including MAPK signaling pathway and PI3K signaling pathway. KRAS protein plays an important role in regulating cell growth, proliferation, differentiation, apoptosis and other life activities (Cell, 2012, 149: 656-670). KRAS gene mutation frequency is high. As early as decades ago, researchers have identified KRAS as an important therapeutic target for cancer. Unfortunately, as a GTPase, KRAS has a high binding affinity with GTP at the picomolar level. Moreover, the concentration of KRAS substrate GTP in cells is high, which makes it difficult to develop nucleotide competitive inhibitors that directly target the GTP pocket (Cancer Cell, 2016, 29 (3): 251-253). In addition, the surface of KRAS protein is very smooth, and it is difficult to find other small molecule binding pockets except the GTP binding site (Nat Chem Biol, 2014, 10(8): 613-622). Therefore, in the past few decades, the development of drugs targeting KRAS has been repeatedly frustrated. KRAS is also one of the most famous "undruggable" targets.

[0005] In 2013, a research team led by Professor Kevan M. Shokat of the University of California, San Francisco (UCSF) discovered a series of compounds that specifically bind to KRAS G12C mutants (Nature, 2013, 503: 548-551). Through crystallographic studies of the complexes of compounds bound to KRAS G12C mutants, the team discovered that there is a "pocket" on the KRAS G12C mutant protein that can bind to small molecule drugs. After binding to this "pocket", small molecule compounds can "lock" the KRAS G12C mutant in an inactive conformation. Based on this research result, several targeted KRAS G12C inhibitors have been promoted to clinical research and application. AMG510 is the first oral KRAS G12C inhibitor to enter clinical research. The research results of Canon and other research groups show that AMG510 can significantly reduce the size of KRAS G12C mutant tumors (Nature, 2019, 575: 217-223).

[0006] As of today, there are several KRAS G12C targeted drugs under development that are in the clinical research stage, including: Adagrasib (MRTX849), Sotorasib (AMG510), JNJ-74699157 (ARS-3248), LY3499446 and BI 1701963. However, KRAS G12C inhibitors still have some fatal weaknesses: (1) Singleness: As mentioned above, KRAS can be divided into multiple types of mutations. The KRAS inhibitors currently under development only target G12C mutations, and have no inhibitory effect on other mutation types (G12A, G12D, G12R, G12S and G12V); (2) Drug resistance: Although many cancer patients treated with KRAS G12C inhibitors have observed clinical benefits, existing clinical data show that the above-mentioned KRAS G12C inhibitors have serious drug resistance problems during clinical use, and multi-drug combination therapy is required to effectively inhibit tumor proliferation. Therefore, exploring new targets for inhibiting KRAS and designing and developing highly effective inhibitors for all KRAS subtypes have always been the hot spots and difficulties in the research of new KRAS inhibitors. Summary of the invention

[0007] The present invention aims to solve one of the technical problems in the related art at least to a certain extent. To this end, one object of the present invention is to provide a compound shown in Formula I or a stereoisomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound shown in Formula I for use in the preparation of a drug for treating cancer. The compound provided by the present invention can effectively stabilize the G-quadruplex structure of the KRAS promoter region, thereby downregulating the transcription level of KRAS mRNA and effectively inhibiting the expression of KRAS protein, and can be used as a KRAS inhibitor. At the same time, the compound can be used as a highly effective inhibitor for all KRAS mutation types, and can inhibit the proliferation of tumor cells.

[0008] To this end, the first aspect of the present invention provides a use of a compound in the preparation of a drug for treating cancer. According to an embodiment of the present invention, the cancer is associated with upregulation of KRAS gene expression or KRAS gene mutation,

[0009] The compound is a compound of Formula I or a stereoisomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound of Formula I:

[0010]

[0011] Among them, R 1 , R 2 , R 3 , R 4are independently selected from optionally substituted C 1-10 Alkoxy, and R 1 , R 2 , R 3 , R 4 At least one of them is a hydroxyl group.

[0012] The KRAS promoter region contains a nuclease hypersensitive element (NHE) located upstream of the transcription start site. The G-rich sequence 32R (5′-GGAGGGGGACAAGGGAGAAGGGTGTGGCGGGA-3′) in the KRAS NHE region can fold to form an intramolecular parallel G-quadruplex under human physiological conditions. Small molecule ligands that can stabilize the KRAS G-quadruplex can effectively regulate the KRAS signaling pathway and are new hot targets for the development of new KRAS inhibitors.

[0013] The inventors found that the compound shown in Formula I can effectively stabilize the G-quadruplex structure in the KRAS promoter region, thereby downregulating the transcription level of KRAS mRNA and effectively inhibiting the expression of KRAS protein. The target of this series of compounds is the guanine (G)-rich DNA sequence in the promoter region of the KRAS gene. Regardless of whether there is a mutation at exon 12, the active ligand molecule can interact with the G-quadruplex structure, thereby downregulating the transcription level of the KRAS gene and inhibiting the expression of the KRAS protein.

[0014] Compared with existing inhibitors that only target KRAS G12 C mutations, this series of compounds has inhibitory effects on tumor cells of all KRAS mutation types. It is very promising to be developed into a new type of KRAS protein inhibitor and has good application prospects.

[0015] According to an embodiment of the present invention, in the compound represented by formula I: R 1 , R 2 , R 3 , R 4 are independently selected from optionally substituted C 1-6 Alkoxy.

[0016] According to an embodiment of the present invention, in the compound represented by formula I: R 1 , R 2 , R 3 , R 4 are independently selected from C 1-4 Alkoxy.

[0017] When R 1 , R 2 , R 3 , R4 are independently selected from C 1-4 Alkoxy, and R 1 , R 2 , R 3 , R 4 When at least one of the groups is a hydroxyl group, the compound can effectively stabilize the G-quadruplex structure of the KRAS promoter region, effectively inhibit the transcription of KRAS mRNA and have a significant inhibitory effect on the expression of KRAS protein.

[0018] According to an embodiment of the present invention, in the compound represented by formula I: R 1 , R 2 , R 3 , R 4 are independently selected from C 1-4 Alkoxy and the C 1-4 The alkoxy group is substituted with one or more halogens.

[0019] According to an embodiment of the present invention, the KRAS gene mutations include G12A, G12C, G12D, G12R, G12S and G12V mutations.

[0020] According to an embodiment of the present invention, the cancer includes at least one selected from pancreatic cancer, lung cancer and colon cancer. A second aspect of the present invention provides the use of a compound in the preparation of a drug for treating cancer. According to an embodiment of the present invention, the cancer is associated with upregulation of KRAS gene expression or KRAS gene mutation, and the compound is a compound shown in the following formula II-VII or a stereoisomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound shown in formula II-VII:

[0021]

[0022] The compound according to the embodiment of the present invention can effectively stabilize the G-quadruplex structure of the KRAS promoter region, inhibit the transcription of KRAS mRNA and have a significant inhibitory effect on the expression of KRAS protein, and also have an inhibitory effect on the proliferation of tumor cells of all KRAS mutation types.

[0023] According to an embodiment of the present invention, the KRAS gene mutations include G12A, G12C, G12D, G12R, G12S and G12V mutations.

[0024] According to an embodiment of the present invention, the cancer includes at least one selected from pancreatic cancer, lung cancer and colon cancer. The third aspect of the present invention provides the use of the compound in the use described in the first aspect or the second aspect in the preparation of a KRAS inhibitor. According to an embodiment of the present invention, the present invention provides the use of a compound in the preparation of a KRAS inhibitor, the compound being a compound of formula I or a stereoisomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound of formula I:

[0025]

[0026] Among them, R 1 , R 2 , R 3 , R 4 are independently selected from optionally substituted C 1-10 Alkoxy, and R 1 , R 2 , R 3 , R 4 At least one of them is a hydroxyl group.

[0027] According to an embodiment of the present invention, in the compound represented by formula I: R 1 , R 2 , R 3 , R 4 are independently selected from optionally substituted C 1-6 Alkoxy.

[0028] According to an embodiment of the present invention, in the compound represented by formula I: R 1 , R 2 , R 3 , R 4 are independently selected from C 1-4 Alkoxy.

[0029] According to an embodiment of the present invention, in the compound represented by formula I: R 1 , R 2 , R 3 , R 4 are independently selected from C 1-4 Alkoxy and the C 1-4 The alkoxy group is substituted with one or more halogens.

[0030] According to an embodiment of the present invention, the present invention provides the use of a compound in the preparation of a KRAS inhibitor, wherein the compound is a compound represented by the following formula II-VII or a stereoisomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound represented by formula II-VII:

[0031]

[0032] The fourth aspect of the present invention provides a KRAS inhibitor. According to an embodiment of the present invention, the KRAS inhibitor comprises the compound for the use described in the first aspect or the second aspect.

[0033] According to an embodiment of the present invention, the KRAS inhibitor provided by the present invention contains a compound represented by Formula I or a stereoisomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound represented by Formula I:

[0034]

[0035] Among them, R 1 , R 2 , R 3 , R 4 are independently selected from optionally substituted C 1-10 Alkoxy, and R 1 , R 2 , R 3 , R 4 At least one of them is a hydroxyl group.

[0036] According to an embodiment of the present invention, in the compound represented by formula I: R 1 , R 2 , R 3 , R 4 are independently selected from optionally substituted C 1-6 Alkoxy.

[0037] According to an embodiment of the present invention, in the compound represented by formula I: R 1 , R 2 , R 3 , R 4 are independently selected from C 1-4 Alkoxy.

[0038] According to an embodiment of the present invention, in the compound represented by formula I: R 1 , R 2 , R 3 , R 4 are independently selected from C 1-4 Alkoxy and the C 1-4 The alkoxy group is substituted with one or more halogens.

[0039] According to an embodiment of the present invention, the KRAS inhibitor provided by the present invention contains the following compounds represented by Formula II-VII or stereoisomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, pharmaceutically acceptable salts or prodrugs of the compounds represented by Formula II-VII:

[0040]

[0041] The fifth aspect of the present invention provides use of the compound in the use described in the first aspect or the second aspect in the preparation of a drug for inhibiting tumor cell proliferation.

[0042] According to an embodiment of the present invention, the tumor cell is a KRAS mutant tumor cell.

[0043] According to an embodiment of the present invention, the tumor cell is selected from at least one of lung cancer cells, colon cancer cells and pancreatic cancer cells.

[0044] According to an embodiment of the present invention, the present invention provides the use of a compound in the preparation of a drug for inhibiting tumor cell proliferation, wherein the compound is a compound represented by Formula I or a stereoisomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound represented by Formula I:

[0045]

[0046] Among them, R 1 , R 2 , R 3 , R 4 are independently selected from optionally substituted C 1-10 Alkoxy, and R 1 , R 2 , R 3 , R 4 At least one of them is a hydroxyl group.

[0047] According to an embodiment of the present invention, in the compound represented by formula I: R 1 , R 2 , R 3 , R 4 are independently selected from optionally substituted C 1-6 Alkoxy.

[0048] According to an embodiment of the present invention, in the compound represented by formula I: R 1 , R 2 , R 3 , R 4 are independently selected from C 1-4 Alkoxy.

[0049] According to an embodiment of the present invention, in the compound represented by formula I: R 1 , R 2 , R 3 , R 4 are independently selected from C 1-4 Alkoxy and the C 1-4 The alkoxy group is substituted with one or more halogens.

[0050] According to an embodiment of the present invention, the present invention provides the use of a compound in the preparation of a drug for inhibiting tumor cell proliferation, wherein the compound is a compound represented by the following formula II-VII or a stereoisomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound represented by formula II-VII:

[0051]

[0052] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] The above and / or additional aspects and advantages of the present invention will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

[0054] Figure 1 The results of measuring the mRNA transcription level of the KRAS gene by fluorescence quantitative PCR in Example 2 of the present invention are shown, wherein Figure A and Figure B respectively show the KRAS mRNA levels in the blank group, compound III, IV and VII groups incubated for 24 hours and 72 hours, respectively, ***, p<0.001;

[0055] Figure 2 The Western blot results of the blank group, control group, compound III, compound IV and compound VII groups in Example 3 of the present invention are shown;

[0056] Figure 3 The grayscale graph of Western blot results of the blank group, control group, compound III, compound IV and compound VII groups in Example 3 of the present invention is shown, *, p<0.05. DETAILED DESCRIPTION

[0057] The present invention is described below with reference to specific embodiments. It should be noted that these embodiments are merely illustrative and do not limit the present invention in any way.

[0058] Unless otherwise specified, the reagents used in the experiments of the embodiments are commercially available.

[0059] In addition, the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of the features. In the description of the present disclosure, "plurality" means at least two, such as two, three, etc., unless otherwise clearly and specifically defined.

[0060] use

[0061] According to a specific embodiment of the present invention, the present invention provides the use of a compound in the preparation of a drug for treating cancer, wherein the cancer is associated with up-regulation of KRAS gene expression or KRAS gene mutation, and the compound is a compound of Formula I or a stereoisomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound of Formula I;

[0062]

[0063] Among them, R 1 , R 2 , R 3 , R 4 are independently selected from optionally substituted C 1-10 Alkoxy, and R 1 , R 2 , R 3 , R 4 At least one of them is a hydroxyl group (for example, R 1 , R 2 , R 3 , R 4 According to another embodiment of the present invention, in the compound represented by formula I: R 1 , R 2 , R 3 , R 4 are independently selected from optionally substituted C 1-6 Alkoxy, or R 1 , R 2 , R 3 , R 4 are independently selected from optionally substituted C 1-6 Alkoxy, or, R 1 , R 2 , R 3 , R 4 are independently selected from C 1-4 Alkoxy and the C 1-4 The alkoxy group is substituted by one or more halogens, wherein the halogens are fluorine, chlorine, bromine or iodine.

[0064] According to an embodiment of the present invention, the types of KRAS gene mutations include but are not limited to G12A, G12C, G12D, G12R, G12S and G12V mutations, and other forms of KRAS gene mutations are also within the scope of the present invention.

[0065] According to a preferred embodiment of the present invention, the present invention provides the use of a compound of formula I in the preparation of a drug for treating cancer, wherein the cancer is associated with up-regulation of KRAS gene expression or KRAS gene mutation, wherein R 1 , R 2 , R 3 , R 4 are independently selected from C 1-4 Alkoxy, and R 1 , R 2 , R 3 , R 4 At least one of them is a hydroxyl group, for example, R 1 , R 2 , R 3 , R 4 Any one of them is hydroxyl group, and the other three are C 1-4 Alkoxy; or R 1 , R 2 , R 3 , R 4 Any two of them are hydroxyl groups, and the other two are C 1-4 Alkoxy; or R 1 , R 2 , R 3 , R 4 Any three of them are hydroxyl groups, and the other one is C 1-4 Alkoxy; or R 1 , R 2 , R 3 , R 4 All are hydroxyl groups, and the KRAS gene mutation types include G12A, G12C, G12D, G12R, G12S and G12V mutations.

[0066] According to a specific embodiment of the present invention, the present invention provides the use of a compound of formula II-VII or a stereoisomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of a compound of formula II-VII in the preparation of a drug for treating cancer, wherein the cancer is associated with up-regulation of KRAS gene expression or KRAS gene mutation,

[0067]

[0068] Pharmaceutical composition

[0069] According to one embodiment of the present invention, the pharmaceutical composition provided by the present invention comprises: the aforementioned compound; and a pharmaceutically acceptable excipient, carrier, excipient, solvent or a combination thereof. The pharmaceutical composition can be used as a highly effective inhibitor for all KRAS mutation types, and can inhibit the proliferation of tumor cells. KRAS mutation types include but are not limited to G12A, G12C, G12D, G12R, G12S and G12V mutations.

[0070] According to one embodiment of the present invention, the compound represented by Formula I-VII or the stereoisomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound represented by Formula I-VII can be used to prepare a drug for treating cancer, wherein the cancer is associated with upregulation of KRAS gene expression or KRAS gene mutation.

[0071] According to one embodiment of the present invention, the types of cancer associated with KRAS gene mutations also refer to cancers caused by KRAS gene mutations, including but not limited to pancreatic cancer, lung cancer and colon cancer, non-small cell lung cancer, lung cancer, lung adenocarcinoma, adenocarcinoma, etc. All cancers associated with KRAS gene mutations reported in the art are included in the protection scope of the present invention. KRAS gene mutation types include but are not limited to G12A, G12C, G12D, G12R, G12S and G12V mutations, and other KRAS gene mutation forms are also included in the scope of the present invention. The compounds provided by the present invention can have inhibitory effects on various types of KRAS mutations, overcome the defects of existing KRAS inhibitors that only have inhibitory effects on single mutations, and can effectively inhibit all KRAS mutation subtypes.

[0072] According to one embodiment of the present invention, the upregulation of KRAS gene expression refers to an increase in the mRNA level of the KRAS gene or an increase in the expression of the KRAS protein. The compounds provided by the present invention can stabilize the G-quadruplex structure of the KRAS gene promoter, thereby downregulating the transcription level of KRAS mRNA and effectively inhibiting the expression of the KRAS protein.

[0073] According to one embodiment of the present invention, the compound shown in Formula I-VII or the stereoisomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the compound shown in Formula I-VII can be used to prepare a drug for inhibiting tumor cell proliferation. The tumor cell is a KRAS mutant tumor cell, and the KRAS mutant tumor cell includes but is not limited to non-small cell lung cancer cells, lung cancer cells, lung adenocarcinoma cells, adenocarcinoma cells, such as human non-small cell lung cancer H1792 cells, H358 cells, H23 cells, human lung adenocarcinoma H1437 cells, H1299 cells, human lung cancer Calu-1 cells, and human adenocarcinoma A549 cells.

[0074] According to one embodiment of the present invention, the pharmaceutical composition provided by the present invention can be introduced into the body such as muscle, intradermal, subcutaneous, intravenous, mucosal tissue by injection, spraying, nasal drops, eye drops, penetration, absorption, physical or chemical mediation; or introduced into the body after being mixed or wrapped with other substances, but is not limited thereto.

[0075] The drug for treating cancer / inhibiting tumor cell proliferation with the compound represented by Formula I-VII or its pharmaceutically acceptable salt as the active ingredient may further include one or more pharmaceutically acceptable carriers when necessary. The carriers include conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants, etc. in the pharmaceutical field.

[0076] The drug for treating cancer / inhibiting tumor cell proliferation with the compound represented by Formula I-VII or its pharmaceutically acceptable salt as active ingredient can be prepared into various forms such as injection, tablet, powder, granule, capsule, oral liquid, ointment, cream, etc. The above-mentioned various dosage forms of drugs can be prepared according to conventional methods in the pharmaceutical field.

[0077] Definitions and explanations of terms

[0078] Unless otherwise specified, the definitions of groups and terms recorded in the specification and claims of this application, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, definitions of specific compounds in examples, etc., can be arbitrarily combined and combined with each other. The group definitions and compound structures after such combination and combination shall fall within the scope recorded in the specification of this application.

[0079] Unless otherwise defined, all technical terms herein have the same meaning as commonly understood by those skilled in the art to which the subject matter of the claims belongs. Unless otherwise indicated, all patents, patent applications, and public materials cited in the entire text of this article are incorporated herein by reference in their entirety.

[0080] Unless otherwise indicated, conventional methods within the technical scope of the art, such as mass spectrometry, NMR, IR and UV / Vis spectroscopy and pharmacological methods, are used. Unless specifically defined, the terms used herein in the relevant descriptions of analytical chemistry, organic synthetic chemistry, and drugs and medicinal chemistry are known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, drug preparation, formulation and delivery, and in the treatment of patients. For example, the manufacturer's instructions for the use of the kit can be used, or the reaction and purification can be carried out in a manner known in the art or in accordance with the description of this application. The above techniques and methods can usually be implemented according to conventional methods well known in the art, according to the descriptions in the multiple summary and more specific documents cited and discussed in this specification. In this specification, groups and substituents thereof can be selected by those skilled in the art to provide stable structural parts and compounds. When a substituent is described by a conventional chemical formula written from left to right, the substituent also includes a chemically equivalent substituent obtained when the structural formula is written from right to left. For example, -CH 3 O is equivalent to -OCH 3 .

[0081] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0082] The term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable salt of a non-toxic acid or base, including salts of inorganic acids and bases, organic acids and bases. Salts derived from inorganic bases include, but are not limited to, metal salts formed by Al, Ca, Li, Mg, K, Na and Zn; salts derived from organic bases include, but are not limited to, salts of primary, secondary or tertiary amines, including naturally occurring substituted or unsubstituted amines, cyclic amines and basic ion exchange resins, such as ammonium, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, caffeine, procaine, choline, betaine, benzylpenicillin, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, N-ethylpiperidine or organic salts formed by polyamine resins; Salts derived from inorganic and organic acids include, but are not limited to, organic salts formed from sulfuric, phosphoric, nitric, hydrobromic, hydrochloric, formic, acetic, propionic, benzenesulfonic, benzoic, phenylacetic, salicylic, alginic, anthranilic, camphoric, citric, ethylenesulfonic, formic, fumaric, furoic, gluconic, glucuronic, glutamic, glycolic, isethionic, lactic, maleic, malic, mandelic, mucic, pamoic, pantothenic, stearic, succinic, sulfanilic, tartaric, p-toluenesulfonic, malonic, 2-hydroxypropionic, oxalic, glycolic, glucuronic, galacturonic, citric, lysine, arginine, aspartic, cinnamic, p-toluenesulfonic, methanesulfonic, ethanesulfonic, or trifluoromethanesulfonic acids.

[0083] In addition to pharmaceutically acceptable salts, other salts are contemplated by the present invention. They may serve as intermediates in the purification of compounds or in the preparation of other pharmaceutically acceptable salts or may be useful in the identification, characterization or purification of the compounds of the present invention.

[0084] The term "stereoisomer" refers to isomers produced by different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, diastereomers and conformers. The stereochemical definitions and conventions used in the present invention are generally defined in accordance with SP Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994.

[0085] Depending on the choice of raw materials and methods, the compounds of the present invention may exist in the form of one of the possible isomers or a mixture thereof, for example as a pure optical isomer, or as a mixture of isomers, such as a racemic and diastereomeric mixture, depending on the number of asymmetric carbon atoms. When describing optically active compounds, prefixes D and L or R and S are used to represent the absolute configuration of the molecule with respect to the chiral center (or multiple chiral centers) in the molecule. The prefixes D and L or (+) and (–) are symbols for the rotation of plane polarized light caused by the specified compound, where (–) or L indicates that the compound is left-handed. Compounds prefixed with (+) or D are right-handed. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. Specific stereoisomers may also be referred to as enantiomers, and mixtures of the isomers are generally referred to as mixtures of enantiomers. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which may occur when there is no stereoselectivity or stereospecificity in a chemical reaction or process. Many geometric isomers of olefins, C=N double bonds, etc. may also exist in the compounds described herein, and all such stable isomers are contemplated in the present invention. When the compounds described herein contain olefinic double bonds, unless otherwise specified, such double bonds include both E and Z geometric isomers. If the compound contains a disubstituted cycloalkyl group, the cycloalkyl substituents may be in the cis- or trans- configuration.

[0086] When the bonds to the chiral carbon in the formula of the present invention are depicted as straight lines, it should be understood that both the (R) and (S) configurations of the chiral carbon and the enantiomerically pure compounds and mixtures thereof produced therefrom are included within the scope of the general formula. The graphic representation of racemates or enantiomerically pure compounds herein is from Maehr, J. Chem. Ed. 1985, 62: 114-120. Unless otherwise indicated, the absolute configuration of a stereocenter is indicated by a wedge-shaped bond and a dashed bond.

[0087] Optically active (R)- or (S)-isomers can be prepared using chiral synthons or chiral preparations, or resolved using conventional techniques. Compounds of the invention containing asymmetrically substituted carbon atoms can be separated in optically active form or racemic form. Resolution of a racemic mixture of a compound can be carried out by any of a number of methods known in the art. Exemplary methods include fractional recrystallization using a chiral resolution acid that is an optically active salified organic acid. Suitable resolving agents for fractional recrystallization methods are, for example, optically active acids, such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, or various optically active camphorsulfonic acids such as the D and L forms of β-camphorsulfonic acid. Other resolving agents suitable for fractional crystallization methods include α-methyl-benzylamine (e.g., S and R forms or diastereomeric pure forms), 2-phenylglycinol, norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane, etc. The resolution of the racemic mixture can also be carried out by eluting on a column filled with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). High performance liquid chromatography (HPLC) can also be used to carry out supercritical fluid chromatography (SFC). The selection of specific methods and elution conditions, the selection of chromatographic columns can be selected by those skilled in the art according to the structure of the compound and the test results. Further, optically pure starting materials or reagents of known configurations can also be used to obtain any enantiomer or diastereomer of the compounds described in the present invention through stereo organic synthesis.

[0088] The term "tautomer" refers to functional group isomers resulting from the rapid movement of an atom in a molecule between two positions. The compounds of the present invention may exhibit tautomerism. Tautomeric compounds may exist in two or more interconvertible species. Prototropic tautomers arise from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium, and attempts to separate a single tautomer usually produce a mixture whose physicochemical properties are consistent with a mixture of compounds. The position of equilibrium depends on the chemical characteristics within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates; while in phenols, the enol form predominates. The present invention encompasses all tautomeric forms of the compounds.

[0089] The term "solvate" means that the compound of the present invention or a salt thereof includes a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent forces between molecules. When the solvent is water, it is a hydrate.

[0090] The term "prodrug" refers to a compound of the present invention that can be converted into a biologically active compound under physiological conditions or by solvolysis. The prodrug of the present invention is prepared by modifying the functional groups in the compound, and the modification can be removed by conventional operations or in vivo to obtain the parent compound. The prodrug includes a compound formed by connecting a hydroxyl or amino group in the compound of the present invention to any group. When the prodrug of the compound of the present invention is administered to a mammalian subject, the prodrug is cleaved to form a free hydroxyl group and a free amino group, respectively.

[0091] The compounds of the invention may contain unnatural proportions of atomic isotopes on one or more of the atoms that make up the compound. For example, the compounds may be labeled with radioactive isotopes, such as deuterium ( 2 H), tritium ( 3 H), iodine-125( 125 I) or C-14( 14 C) All isotopic variations of the compounds of the present invention, whether radioactive or not, are included within the scope of the present invention.

[0092] The term "C 1 -C 10 "Alkoxy" is understood to mean -O-(C 1 -C 10 alkyl), where "C 1 -C 10 “Alkyl” is understood to mean a linear or branched, saturated, monovalent hydrocarbon radical having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. The alkyl radical is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc. or their isomers. In particular, the radical has 1, 2, 3, 4, 5, 6 carbon atoms (“C 1 -C 6 alkyl), for example methyl, ethyl, propyl, butyl, isopropyl, isobutyl, sec-butyl, tert-butyl, more particularly, the radical having 1, 2 or 3 carbon atoms (“C 1 -C 3 alkyl"), for example methyl, ethyl, n-propyl or isopropyl.

[0093] The term "pharmaceutical composition" refers to a mixture of one or more compounds described herein or their physiologically / pharmaceutically acceptable salts or prodrugs with other chemical components, such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of a compound to an organism.

[0094] With respect to a drug or pharmacologically active agent, the term "effective dose", "effective amount" or "therapeutically effective amount" refers to a sufficient amount of a drug or agent that is non-toxic but can achieve the desired effect. For oral dosage forms of the present invention, an "effective amount" of an active substance in a composition refers to the amount required to achieve the desired effect when used in combination with another active substance in the composition. The determination of the effective amount varies from person to person, depending on the age and general condition of the recipient, and also on the specific active substance. The appropriate effective amount in each case can be determined by a person skilled in the art based on routine experiments.

[0095] The terms "active ingredient," "therapeutic agent," "active substance," or "active agent" refer to a chemical entity that is effective in treating a target disorder, disease, or condition.

[0096] The term "excipient" refers to a pharmaceutically acceptable inert ingredient. Examples of the term "excipient" include, but are not limited to, binders, disintegrants, lubricants, glidants, stabilizers, fillers, and diluents. Excipients can enhance the handling characteristics of a pharmaceutical formulation, i.e., make the formulation more suitable for direct compression by increasing fluidity and / or adhesion. Examples of typical "pharmaceutically acceptable carriers" suitable for the above-mentioned preparations are: sugars, such as lactose, sucrose, mannitol and sorbitol; starches, such as corn starch, tapioca starch and potato starch; cellulose and its derivatives, such as sodium carboxymethylcellulose, ethyl cellulose and methyl cellulose; calcium phosphates, such as dicalcium phosphate and tricalcium phosphate; sodium sulfate; calcium sulfate; polyvinyl pyrrolidone; polyvinyl alcohol; stearic acid; alkaline earth metal salts of stearic acid, such as magnesium stearate and calcium stearate; stearic acid; vegetable oils, such as peanut oil, cottonseed oil, sesame oil, olive oil and corn oil; nonionic, cationic and anionic surfactants; ethylene glycol polymers; fatty alcohols; and cereal hydrolyzed solids and other non-toxic compatible fillers, binders, disintegrants, buffers, preservatives, antioxidants, lubricants, colorants and the like excipients commonly used in pharmaceutical preparations.

[0097] The scheme of the present disclosure will be explained below in conjunction with the examples. Those skilled in the art will appreciate that the following examples are only used to illustrate the present disclosure and should not be considered to limit the scope of the present disclosure. Where specific techniques or conditions are not indicated in the examples, the techniques or conditions described in the literature in this area or the product instructions are used. Where the manufacturers of reagents or instruments are not indicated, they are all conventional products that can be obtained commercially.

[0098] The synthetic routes of the tetrahydropalmatine derivatives shown in formula II, formula IV and formula VII used in the examples are referred to the following documents: Zhong-Ze Ma, Wei Xu, Niels H. Jensen, Bryan L. Roth, Lee-Yuan Liu-Chen and David YW Lee, Molecules 2008, 13, 2303-2312; Ying-Hong Li, Peng Yang, Wei-Jia Kong, Yan-Xiang Wang, Chang-Qin Hu, Zeng-Yan Zuo, Yue-Ming Wang, Hong Gao, Li-Mei Gao, Yan-Chun Feng, Na-Na Du, Ying Liu, Dan-Qing Song and Jian-Dong Jiang, J. Med. Chem. 2009, 52, 492-501. Formula III, formula V and formula VI can all be purchased from Chembest (Shanghai, China).

[0099] Example 1: Evaluation of the Stability of Compounds on KRAS G-quadruplex

[0100] 1. Obtaining KRAS G-quadruplex

[0101] There is a guanine (G)-rich DNA sequence in the promoter region of the KRAS gene, which can form a special DNA structure - G quadruplex (32R) (Nucleic Acids Res., 2020, 48, No. 16, 9336-9345).

[0102] G-quadruplex sequence (32R): 5'-GGAGGGGGACAAGGGAGAAGGGTGTGGCGGGA-3', SEQ ID NO: 1.

[0103] The nucleic acid sequence is artificially synthesized and dissolved in PBS buffer to form a G-quadruplex structure.

[0104] 2. Stability evaluation

[0105] a) Experimental instruments:

[0106] The circular dichroism spectrometer model is Jasco-815 from JASCO Corporation.

[0107] b) Experimental process:

[0108] Before the circular dichroism (CD) experiment, high-purity nitrogen was used for deoxygenation for no less than 5 minutes, and during the experiment, high-purity nitrogen was continuously introduced as a protective gas to ensure that no ozone was produced. Before CD experimental data collection, the baseline was corrected with the corresponding buffer solution. The cuvette used was a double-glossy double-frosted quartz cuvette with an optical path of 1 cm.

[0109] CD experimental parameters: scanning wavelength range 200nm-400nm; scanning speed: 500nm / min; room temperature; slit width 5nm, take the average value of 5 scanning results for data collection. Heating range 25-90℃, heating speed 2℃ / min.

[0110] The CD values ​​at 260 nm were measured to determine the melting points (T) of KRAS G-quadruplexes before and after the addition of different compounds obtained in Example 1. m ) to evaluate the ability of the compounds to stabilize the KRAS G-quadruplex structure.

[0111] c) Experimental results:

[0112] The results of the stability evaluation of compound II-VII on the KRAS G-quadruplex structure are shown in Table 1 below:

[0113] Table 1 Evaluation of the stability of a series of compounds on KRAS G-quadruplex

[0114]

[0115] The above experimental results show that the series of compounds II-VII all have a good stabilizing effect on the KRAS G-quadruplex structure and can be used as KRAS G-quadruplex stabilizers. Among them, compounds III, IV and VII have the best stabilizing effect and are more preferred choices for new KRAS G-quadruplex stabilizers.

[0116] Example 2: Inhibitory effect of compounds on KRAS mRNA transcription

[0117] (1) Experimental parameters and process:

[0118] The above compounds III, IV and VII with the best stabilization effect on KRAS G-quadruplex were used to evaluate the KRAS mRNA transcription inhibition effect. Among them, the human lung cancer cell line A549 was purchased from the Shanghai Institute of Cell Biology, Chinese Academy of Sciences. The experimental process is as follows:

[0119] The control group (blank group) cells (A549 cells without compound + medium) were cultured for 24 and 72 hours, respectively. The experimental group cells were cultured in medium containing compounds (compounds III, IV and VII) with a final concentration of 50 μM for 24 and 72 hours, respectively, and the cells (approximately 5×10 7 The total RNA in the cells was extracted by Trizol method, and then reverse transcribed into cDNA after concentration calibration. The mRNA transcription level of KRAS gene was further determined by fluorescence quantitative PCR.

[0120] Culture medium composition: RPMI 1640 medium (HyClone) containing 10% (v / v) fetal bovine serum (Gibco), 0.1 mg / mL streptomycin (Sigma-Aldrich) and 100 U / mL penicillin (Sigma-Aldrich)

[0121] Fluorescence quantitative PCR primers:

[0122] KRAS-For: 5'-ACACAAACAGGCTCAGGACT-3', SEQ ID NO: 2;

[0123] KRAS-Rev: 5'-TGTCGGATCTCCCTCACCAA-3', SEQ ID NO: 3;

[0124] β-actin-For: 5'-AGCACTGTGTTGGCGTACAG-3', SEQ ID NO: 4;

[0125] β-actin-Rev: 5'-TCCCTGGAGAAGAGCTACGA-3', SEQ ID NO: 5.

[0126] (2) Experimental results:

[0127] Fluorescence quantitative PCR Figure 1 As shown in Figures A and B, Figure A shows that after incubation of A549 cells with compounds III, IV and VII for 24 hours, the mRNA transcription level of the KRAS gene was significantly downregulated, and there was a significant difference with the blank group. This downward trend is proportional to the incubation time. It can be seen that after incubation of A549 cells with compounds III, IV and VII shown in Figure B for 72 hours, the mRNA transcription level of the KRAS gene was further downregulated. Among the three compounds, compound VII has the best inhibitory effect on the transcription level of KRAS.

[0128] Example 3: Inhibitory effect of compounds on KRAS protein expression

[0129] (1) Experimental parameters and process:

[0130] The above compounds III, IV and VII with the best stabilization effect on KRAS G-quadruplex were used to perform an inhibitory effect evaluation experiment on KRAS protein expression to evaluate the inhibitory effect of the compounds on KRAS protein expression. The experimental process is as follows:

[0131] a) Preparation of Western blot experimental samples using A549 cells. The blank control group was cultured with normal A549 cells, the negative control group was cultured in a medium containing 1% v / v DMSO, and the experimental group was cultured in a medium containing different compounds (compounds III, IV and VII) with a final concentration of 50 μM. After incubation with the compound or negative control reagent DMSO for 72 hours, the cells (approximately 5×10 7 indivual);

[0132] b) Cell protein was extracted using RIPA cell lysis buffer, and the total protein content was calibrated using the BCA method;

[0133] c) Prepare SDS-PAGE gel, with the upper layer being 5% concentrated gel and the lower layer being 8% separation gel. Electrophoresis conditions: Constant voltage of concentrated gel at 60V for about 20min; Constant voltage of separation gel at 60-120V, and the stopping time of electrophoresis is determined by pre-staining protein markers;

[0134] d) After electrophoresis, the membrane was transferred using the wet transfer method. The transfer conditions were: 300 mA constant current; 0.22 μm pore size PVDF membrane; transfer time 90-150 min;

[0135] e) Blocking: The membrane was completely immersed in 5% skim milk-TBST and incubated on a horizontal shaker for 1 h (RT);

[0136] f) Primary antibody incubation: dilute the primary antibody with 5% skim milk-TBST, incubate overnight at 4°C on a horizontal shaker, and wash the membrane the next day: wash three times with TBST, 15 min each time;

[0137] g) Secondary antibody incubation: dilute the secondary antibody with 5% skim milk-TBST, goat anti-rabbit IgG (H+L) HRP or goat anti-mouse IgG (H+L) and rabbit anti-goat IgG (H+L) HRP 1:10000, incubate at room temperature for 1 hour. Wash the membrane: wash the membrane 3 times with TBST, 15 minutes each time;

[0138] h) Exposure using ECL luminescent liquid chemiluminescence colorimetry, with exposure time varying from 2 to 30 minutes, using a developing device for development, performing image analysis on the resulting development image, and calculating the grayscale value.

[0139] (2) Experimental results:

[0140] The inhibitory effects of the series of compounds (compounds III, IV and VII) on KRAS protein, such as Figure 2 and 3 As shown, the experimental results show that compared with the blank control group, the series of compounds all have an inhibitory effect on the expression of KRAS protein, especially compounds III and VII significantly inhibit the expression of KRAS protein, indicating that the compounds of the present invention are a new type of KRAS protein inhibitors.

[0141] Example 4: Experimental study on the inhibition of proliferation of different tumor cells by a series of compounds

[0142] (1) Experimental process

[0143] Cell lines: Cell lines include tumor cells with and without G12C mutations, all of which were purchased from Shanghai Union Medical College Cell Research Center. The names and codes of the cells are shown in Table 2 below:

[0144] Table 2

[0145] Rich in G12C mutations No G12C mutation H1792: Human non-small cell lung cancer H1437: Human lung adenocarcinoma cells H358: Human non-small cell lung cancer H1299: Human lung adenocarcinoma cells H23: Human non-small cell lung cancer A549: human adenocarcinoma cells Calu-1: human lung cancer cells

[0146] Screening method: MTT method;

[0147] Duration of action: 72 hours;

[0148] The specific experimental method is as follows (the following uses A549 human adenocarcinoma cells as an example, and the experimental methods for other types of cancer cells are similar):

[0149] Human lung cancer cells A549 growing in logarithmic growth were digested with 0.25% trypsin and then prepared into cell suspensions using DMEM complete culture medium containing 10% (volume percentage) fetal bovine serum. The tumor cells were diluted to 1.0×10 5 pc / mL, and then inoculated into 96-well culture plates, 200 μL per well, and incubated at 37°C, 5% CO 2 Culture in an incubator for 24 h, and then pour out the culture medium from each well.

[0150] The above-mentioned tetrahydropalmatine derivatives (compounds II, III, IV, V, VI, VII) were dissolved in DMSO and diluted to the required concentration with DMEM complete culture medium, the concentration of each tetrahydropalmatine derivative (compound II, III, IV, V, VI, VII) was 50 μg / mL, and the DMSO content was 1% (volume percentage); each tetrahydropalmatine derivative (compound II, III, IV, V, VI, VII) was added to the culture wells of the above-mentioned tumor cells A-549, and 5 wells were inoculated, and 200 μL was added to each well; the control group was respectively added with DMEM complete culture medium containing 1% (volume percentage) DMSO to the culture wells of the above-mentioned tumor cells to a volume of 200 μL in the wells, and 5 wells were also inoculated; the blank group was added with 200 μL of DMEM complete culture medium to the empty culture wells without tumor cells; it was placed at 37°C, 5% CO 2 Culture in an incubator for 72h.

[0151] After 3 days, 20 μL of freshly prepared 5 mg / mL methyl thiazolyl tetrazolium (MTT) solution was added to each well and incubated at 37°C with 5% CO. 2 Continue to culture for 4 h.

[0152] Carefully discard the supernatant, add 200 μL DMSO to each well, and place at room temperature for 15-20 minutes, shaking every 5 minutes.

[0153] The optical density (OD value) at a wavelength of 490 nm was measured on an enzyme-labeled instrument, and the tumor cell growth inhibition rate was calculated according to the formula.

[0154]

[0155] The experiment was repeated three times.

[0156] (2) Experimental results

[0157] The results of the inhibitory effects of various derivatives of tetrahydropalmatine (compounds II, III, IV, V, VI, VII) on different tumor cells are shown in Table 2.

[0158] Table 2 Inhibitory effects of compounds II, III, IV, V, VI, and VII on different human tumor cells

[0159]

[0160] The above results show that the derivatives of tetrahydropalmatine (compounds II, III, IV, V, VI, VII) have good inhibitory effects on various tumor cell lines and have great prospects for medical development.

[0161] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", "some implementation schemes" or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, unless they are contradictory.

[0162] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. Use of the compound in the preparation of a drug for treating cancer, It is characterized in that The cancer is associated with upregulation of KRAS gene expression or KRAS gene mutation, Wherein, the compound is a compound represented by Formula I or a stereoisomer or a pharmaceutically acceptable salt of the compound represented by Formula I: Among them, R 1 , R 2 , R 3 , R 4 are independently selected from methoxy, and R 1 , R 2 , R 3 , R 4 At least one of the molecules is a hydroxyl group, and the cancer is lung cancer.

2. The use according to claim 1, It is characterized in that The KRAS gene mutations include G12A, G12C, G12D, G12R, G12S and G12V mutations.

3. Use of the compound in the preparation of a drug for treating cancer, It is characterized in that The cancer is associated with upregulation of KRAS gene expression or KRAS gene mutation, Wherein, the compound is a compound represented by the following formula II-VII or a stereoisomer or a pharmaceutically acceptable salt of the compound represented by formula II-VII: The cancer is lung cancer.

4. The use according to claim 3, It is characterized in that The KRAS gene mutations include G12A, G12C, G12D, G12R, G12S and G12V mutations.

5. Use of the compound according to any one of claims 1 to 4 in the preparation of a KRAS inhibitor.

6. Use of the compound according to any one of claims 1 to 4 in the preparation of a medicament for inhibiting the proliferation of tumor cells, wherein the tumor cells are KRAS mutant tumor cells, and the KRAS mutant tumor cells are lung cancer cells.

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