Pharmaceutical composition containing PRMT5 inhibitor and krasg12c inhibitor

Abstract

A pharmaceutical composition comprising a protein arginine methyltransferase 5 (PRMT5) inhibitor and a KRASG12C (Kirsten rat sarcoma viral oncogene homolog glycine-to-cysteine) inhibitor. The pharmaceutical composition can be used for treating various cancers, comprising solid tumors. The combined product can be used for treating any number of diseases associated with PRMT5 and / or KRASG12C.

Description

Contains PRMT5 inhibitors and KRAS G12C Pharmaceutical compositions of inhibitors Technical Field

[0001] The present invention belongs to the field of medicine, and specifically relates to a method for protecting PRMT5 inhibitors and KRAS G12C Pharmaceutical compositions of inhibitors. Background Art

[0002] Cancer is a leading cause of death worldwide. Popular treatments, such as chemotherapy and immunotherapy, are limited in that their cytotoxic effects are not restricted to cancer cells and can also cause adverse side effects in normal tissues.

[0003] The rat sarcoma (RAS) proto-oncogene has been identified as an oncogenic driver in cancers such as non-small cell lung cancer (NSCLC) and colorectal cancer (CRC). The RAS family includes three closely related genes that express guanosine triphosphatase (GTPase) enzymes responsible for regulating cell proliferation and survival. The RAS proteins, Kirsten rat sarcoma viral oncogene homolog (KRAS), Harvey rat sarcoma viral oncogene homolog (HRAS), and neuroblastoma RAS viral oncogene homolog (NRAS), can be activated by mutations at codons 12, 13, or 61, leading to cancer in humans. Different tumor types are associated with mutations in certain isoforms of RAS1, with KRAS being the most frequently mutated isoform in most cancers. Although the role of KRAS mutations in human cancers has been known for decades, anti-cancer therapies targeting KRAS mutations have been unsuccessful, primarily because the protein was considered refractory to inhibition by small molecules. Until recently, the U.S. Sotorasib, marketed under the name , is the first inhibitor of the RAS GTPase family and is indicated for adult patients with locally advanced or metastatic NSCLC harboring a KRAS G12C mutation who have received at least one prior systemic therapy. While sotorasib monotherapy appears to produce significant anti-tumor activity, combination therapies are desirable to further improve the treatment options available to patients.

[0004] PRMT5 is a type II arginine methyltransferase that regulates important cellular functions, including cell cycle progression, apoptosis, and the DNA damage response, by symmetrically dimethylating proteins involved in transcription and signal transduction. However, data from genome-wide genetic perturbation screens using shRNA revealed a selective requirement for PRMT5 activity in MTAP-deleted cancer cell lines (Kruykov et al., 2016; Marjon et al., 2016; Markarov et al., 2016). The accumulation of MTA caused by MTAP loss in these cell lines partially inhibits PRMT5, making these cells selectively sensitive to additional PRMT5 inhibition.

[0005] Certain PRMT5 inhibitors have been developed, but they do not show selectivity for MTAP-deficient cancer cell lines. This lack of selectivity could be explained by the inhibitors’ mechanism of action, as they are either SAM-uncompetitive or SAM-competitive inhibitors and therefore independent of MTAP (Kruykov et al., 2016; Marjon et al., 2016 and Markarov et al., 2016).

[0006] By using inhibitors that bind PRMT5 non-competitively or cooperatively with MTA, selectivity for MTAP-deficient / MTA-accumulating cells can be improved. PRMT5 inhibitors that bind non-competitively or cooperatively with MTA will exhibit increased binding to PRMT5 in the presence of MTA compared to the binding of the same inhibitor in the absence of MTA. Consequently, such inhibitors will bind with significantly greater potency in the presence of high concentrations of MTA and, therefore, lead to preferential inhibition of PRMT5 in MTA-accumulating cells relative to normal cells.

[0007] Despite many recent advances in cancer treatment, there remains a need for more effective and / or enhanced treatments for those affected by cancer. Summary of the Invention

[0008] In order to solve the technical problem of the present invention, the present invention provides a pharmaceutical composition comprising a PRMT5 inhibitor having a first active substance and a KRAS inhibitor having a second active substance. G12C Inhibitor compound, wherein the PRMT5 inhibitor of the first active substance has the following structure of formula (I) or formula (II):

[0009] Wherein, in formula (I) or formula (II),

[0010] R1 is selected from H, halogen, C1-C6 alkyl, halo(C1-C6 alkyl), CN;

[0011] R2 is selected from H, C1-C6 alkyl, halo(C1-C6 alkyl), C3-C6 cycloalkyl;

[0012] R3 represents H, halogen, C1-C6 alkyl, halo(C1-C6 alkyl), halo(C1-C6 alkoxy), or SF5;

[0013] In formula II, R4 represents hydrogen or C1-C6 alkyl; X represents CR5 or N;

[0014] Wherein, R5 represents hydrogen, halogen, C1-C6 alkyl, halo(C1-C6 alkyl), hydroxyl, -NH2, or CN.

[0015] In the preferred technical solution of the present disclosure, wherein, in the structure of formula (I) or formula (II):

[0016] R1 is selected from hydrogen, halogen, C1-C6 alkyl, halo(C1-C6 alkyl);

[0017] R2 is selected from hydrogen, C1-C6 alkyl, halo(C1-C6 alkyl), C3-C6 cycloalkyl;

[0018] R3 represents hydrogen, halogen, halo(C1-C6 alkyl), halo(C1-C6 alkoxy), or SF5;

[0019] R4 represents hydrogen or methyl;

[0020] In formula II, X represents CH or N.

[0021] In the preferred technical solution of the present disclosure, R1 is selected from hydrogen or fluorine.

[0022] In the preferred technical solution of the present disclosure, R2 is selected from cyclopropyl or methyl.

[0023] In the preferred technical solution of the present disclosure, R3 represents CF3.

[0024] In the preferred technical solution of the present disclosure, R4 represents hydrogen or methyl.

[0025] In the preferred technical solution of the present disclosure, X represents N.

[0026] In the preferred technical solution of the present disclosure, the PRMT5 inhibitor of the first active substance is selected from the following compounds or any combination thereof:

[0027] In the preferred technical solution disclosed herein, the KRAS G12CThe inhibitor is selected from any one of the following compounds or their pharmaceutical salts: Adagrasib Sotorasib( and ), Divarasib (GDC-6036), Glecirasib (JAB-21822), LY3537982 (Eli Lilly and company), Garsorasib (D-1553), HBI-2438 (Huyabio International), Opnurasib (JDQ443), HS-10370 (Jiangsu Hansoh pharmaceutical company), IBI-351 (GFH925), BI-1823911 (Boehringer Ingelheim), JNJ-74699157 (ARS3248, Johnson & Johnson), RM-018 (Revolution Medicines), ARS-2102, RMC-4998, ASP2453, BI-2865, AZD4747, BI-0474, MK-108, BEBT-607, YL-15293, D3S-001, ABSK071, LY3499446, RMC-6291, SY-5933, GEC-255, RMC-4630 (SAR442720), BPI-421286, or any combination thereof.

[0028] In the preferred technical solution disclosed herein, the KRAS G12C The inhibitor was selected from the Protac degrader LC2 (CAS: 2502156-03-6).

[0029] In the preferred technical solution disclosed herein, the KRAS G12C Inhibitor selected from Adagrasib Sotorasib( and ), Opnurasib (JDQ443), LY3537982 (Eli Lilly and company) or a pharmaceutically acceptable salt thereof.

[0030] In addition, the present disclosure also provides a method for treating cancer or tumor, comprising administering the pharmaceutical composition of the present invention to an individual in need thereof.

[0031] In the preferred technical scheme of the present disclosure, the tumor or cancer is selected from: glioblastoma multiforme, brain cancer, prostate cancer, pancreatic cancer, mantle cell lymphoma, non-Hodgkin's lymphoma and diffuse large B-cell lymphoma, acute myeloid leukemia, acute lymphoblastic leukemia, multiple myeloma, non-small cell lung cancer, small cell lung cancer, breast cancer, triple-negative breast cancer, gastric cancer, colorectal cancer, ovarian cancer, bladder cancer, hepatocellular carcinoma, esophageal cancer, bile duct cancer, mesothelioma, laryngeal cancer, melanoma, malignant peripheral nerve sheath tumor, osteosarcoma, myxochondrosarcoma, soft tissue sarcoma, oropharyngeal squamous cell carcinoma, chronic myeloid leukemia, epidermal squamous cell carcinoma, nasopharyngeal carcinoma, neuroblastoma, endometrial cancer, head and neck cancer and cervical cancer.

[0032] As will be understood by those of ordinary skill in the art, in any embodiment disclosed herein, any feasible combination of a compound of formula (I) or a pharmaceutically acceptable salt, solvate or prodrug thereof and a compound of formula (II) or a pharmaceutically acceptable salt, solvate or prodrug thereof is included in the present invention, as long as such combination is capable of producing some synergistic effect in the treatment of a subject in need of such treatment.

[0033] When any compound is used in the present invention, it includes any pharmaceutically acceptable form thereof, including but not limited to isomers, tautomers, salts, solvates, polymorphs, prodrugs, etc. It should be understood that the term "compound" includes any and all such forms, whether or not explicitly stated, although sometimes only certain terms are explicitly stated, such as "salt" and "prodrug".

[0034] Unless expressly defined otherwise, all terms used herein have the ordinary meaning as would be interpreted or understood by one of ordinary skill in the art.

[0035] The terms "a", "an" or "the" used herein refer to both the singular and the plural forms. Generally, when a singular or plural form of a noun is used, it refers to both the singular and the plural forms of the noun.

[0036] When the term "about" is applied to a parameter, it means that the parameter can vary within ±10%, preferably within ±5%, including any number from the lower limit to the upper limit. When the term "about" is applied to a range, it applies to both the lower and upper limits of the range. As will be understood by those skilled in the art, when a parameter is not critical, a number is generally given for illustrative purposes only and is not limiting.

[0037] "Alkoxy" refers to a group -OR, where R is alkyl as defined herein. Representative examples include methoxy, ethoxy, propoxy, isopropoxy, sec-butoxy, tert-butoxy, and the like.

[0038] "Alkyl" refers to a group derived from a straight or branched chain saturated hydrocarbon by removing a hydrogen from one of the saturated carbons. Alkyl groups preferably contain 1 to 8 carbon atoms, sometimes preferably 1 to 6 carbon atoms, and sometimes even more preferably 1 to 4 carbon atoms. Representative examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, and the like. "Lower alkyl," "lower alkoxy," or "lower haloalkyl" refers to an alkyl or alkyl moiety having one to four, sometimes preferably one to three or one to two carbon atoms.

[0039] As used herein, the term "cyano" refers to -CN.

[0040] The term "cycloalkyl" as used herein refers to a group derived from a monocyclic saturated carbocyclic ring by removing a hydrogen atom from the saturated carbocyclic ring, preferably having 3 to 8, more preferably 3 to 6 carbon atoms. Representative examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclopentyl, and cyclohexyl.

[0041] As used herein, the terms "halo" and "halogen" refer to F, Cl, Br, or I.

[0042] The term "haloalkyl" as used herein refers to an alkyl group substituted with at least one halogen atom. A haloalkyl group can be an alkyl group in which all hydrogen atoms are substituted with halogens. Representative examples of haloalkyl groups include, but are not limited to, trifluoromethyl, fluoromethyl, difluoromethyl, bromomethyl, 1-chloroethyl, perchloroethyl, 2-fluoroethyl, etc.

[0043] The term "heterocyclyl" as used herein refers to a 3 to 10-membered monocyclic or bicyclic non-aromatic group containing one or more, preferably 1 to 3, heteroatoms independently selected from nitrogen (N), oxygen and sulfur (S, S(O) or S(O)2) in the non-aromatic ring. The heterocyclyl of the present disclosure can be connected to the parent molecular moiety through a carbon atom or a nitrogen atom in the group. The heterocyclyl group can be saturated or unsaturated, for example, containing one or more double bonds in the ring. Unless otherwise stated, the valence of the group can be located on any atom of any ring within the group where the valence rules permit. Examples may include, but are not limited to, azetidinyl, pyrrolidinyl, 2-oxopyrrolidinyl, 2,5-dihydro-1H-pyrrolyl, piperidinyl, 4-piperidinyl, morpholinyl, piperazinyl, 2-oxopiperazinyl, tetrahydropyranyl, tetrahydrofuranyl, 2-oxopiperidinyl, thiomorpholinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, etc.

[0044] When any group, such as "cycloalkyl" or "heterocyclyl" is referred to as "substituted or unsubstituted" or "optionally substituted", unless otherwise specified, it means that the group is substituted or not substituted by 1 to 5, sometimes preferably 1 to 3 or 1 to 2 substituents independently selected from halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl and cyano.

[0045] The term "solvate" as used herein refers to a physical association of a compound of the invention with one or more, preferably one to three, solvent molecules (whether organic or inorganic). This physical association includes hydrogen bonding. In some cases, the solvate is capable of separation, for example when one or more, preferably one to three, solvent molecules are incorporated into the crystal lattice of a crystalline solid. Exemplary solvates include, but are not limited to, hydrates, ethanolates, methanolates, and isopropanolates. Solvation methods are generally known in the art.

[0046] "Prodrugs" refer to compounds that can be converted in vivo to produce the active parent compound under physiological conditions, such as by hydrolysis in the blood. Common examples include, but are not limited to, ester and amide forms of compounds having an active form with a carboxylic acid moiety. Amides and esters of the compounds of the present invention can be prepared according to conventional methods. In particular, in the present invention, prodrugs can also be formed by acylation of the amino group or nitrogen atom in the heterocyclyl ring structure, where the acyl group can hydrolyze in vivo. Such acyl groups include, but are not limited to, C1-C6 acyl groups, preferably C1-C4 acyl groups, more preferably C1-C2 (formyl or acetyl) groups, or benzoyl groups.

[0047] As used herein, the term "subject" refers to a human or other mammal, such as a monkey, dog, cat, horse, etc. The term is intended to encompass and is sometimes interchangeable with "patient."

[0048] As used herein, the terms "administering" or "administering" refer to providing a compound or pharmaceutical composition to a subject having or at risk for a disease or condition to be treated or prevented.

[0049] Any route of administration is suitable for the present invention. In one embodiment, the compounds of the present invention can be administered to a subject in a solid dosage form such as a tablet, capsule, or the like. In one embodiment, the compounds of the present invention can be administered to a subject by intravenous injection. In another embodiment, the compounds of the present invention can be administered to a subject by any other suitable systemic delivery method, such as oral, parenteral, intranasal, sublingual, rectal, or transdermal administration.

[0050] As used herein, the term "therapeutically effective amount" refers to that amount of a compound or composition that will elicit the desired or intended biological or medical response in a subject that is being sought by a physician, veterinarian, or researcher. The therapeutically effective amount of the compound and the specific pharmaceutically acceptable carrier will vary depending on, for example, the age, weight, sex of the subject, the mode of administration, and the disease or condition being treated.

[0051] The term "pharmaceutically acceptable" when used before a compound, salt, prodrug, composition, or carrier means that such compound, salt, prodrug, composition, or carrier is suitable for administration to a subject for treatment without causing intolerable side effects to the subject considering the desired treatment.

[0052] As used herein, the term "pharmaceutically acceptable carrier" refers to a substance that is compatible with the compounds used in the present invention and can be used to administer the compounds in the methods of the present invention, and is preferably non-toxic, or inert and pharmaceutically acceptable. Pharmaceutically acceptable carriers can be solid, liquid or gaseous substances, including any and all dry powders, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents, absorption delaying agents, and the like. Examples of such carriers include oils such as corn oil, buffers such as phosphate buffered saline (PBS), saline, polyethylene glycol, glycerol, polypropylene glycol, dimethyl sulfoxide, amides such as dimethylacetamide, proteins such as albumin, detergents such as Tween 80, monosaccharides and oligosaccharides such as glucose, lactose, cyclodextrins, starch, and the like.

[0053] As described herein, some embodiments of the compounds of the present invention can contain basic functional groups, such as amino or alkylamino, and therefore can form pharmaceutically acceptable salts with pharmaceutically acceptable acids. In this respect, the term "pharmaceutically acceptable salts" refers to relatively nontoxic inorganic and organic acid addition salts of the compounds of the present invention. These salts can be prepared on site during the administration of carriers or dosage form production processes, or by reacting the purified compounds of the present invention in free alkali form with suitable organic or inorganic acids alone, and separating the salts so formed in subsequent purification processes to prepare. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, toluenesulfonate, citrate, maleate, fumarate, succinate.

[0054] The formulations used in the present invention may also contain stabilizers, preservatives, buffers, antioxidants or other additives known to those skilled in the art. The use of such media and agents for pharmaceutically active substances is well known in the art.

[0055] The terms "synergistic" and the like as used herein refer to an effect caused by a combination of two or more agents that is greater than the cumulative effect of the two or more agents used alone. This synergistic effect of combination therapy includes higher efficacy, lower side effects, or both. In some embodiments, the synergistic effect includes a significant reduction in the side effects of the two therapeutic inhibitors due to a reduction in the dosage of the two therapeutic inhibitors, while the overall therapeutic efficacy remains at approximately the same or improved levels. In some embodiments, the synergistic effect includes a significant improvement in the efficacy of inhibiting cancer cell proliferation, while the side effects caused by the two drugs remain at approximately the same or lower levels. The synergistic effect allows the use of a lower dose of a single drug to effectively treat the disease. In general, the synergistic combination of two or more drugs can lead to improvements in disease treatment compared to monotherapy.

[0056] Combination therapy can allow the use of a lower dose of a first therapeutic agent, such as a PRMT5 inhibitor, or a second therapeutic agent, such as a MAT2A inhibitor, or lower doses of both therapeutic agents than would normally be required when either agent is used alone. The present invention encompasses any and all such "synergistic" effects.

[0057] The pharmaceutical composition may comprise a PRMT5 inhibitor and a KRAS G12C The total amount of the inhibitor is 0.01% to 99% by weight of the total composition, preferably 0.1% to 80% by weight of the total composition, more preferably 0.1% to 50% by weight of the total composition. G12C The weight ratio between the inhibitors may be in the range of 1:20 to 20:1, sometimes preferably 1:15 to 15:1, sometimes more preferably 1:10 to 10:1.

[0058] For systemic administration, the daily dosage range for adult human treatment of a PRMT5 inhibitor is about 0.01 to about 150 mg / kg, preferably about 0.05 to about 100 mg / kg, and sometimes more preferably about 0.1 to about 50 mg / kg.

[0059] The present disclosure provides pharmaceutical compositions that can be used to treat and / or prevent various cancers that may include or exclude the following cancers: glioblastoma multiforme, brain cancer, prostate cancer, pancreatic cancer, mantle cell lymphoma, non-Hodgkin lymphoma and diffuse large B-cell lymphoma, acute myeloid leukemia, acute lymphoblastic leukemia, multiple myeloma, non-small cell lung cancer, small cell lung cancer, breast cancer, triple-negative breast cancer, gastric cancer, colorectal cancer, ovarian cancer, bladder cancer, hepatocellular carcinoma, esophageal cancer, bile duct cancer, mesothelioma, laryngeal cancer, melanoma, malignant peripheral nerve sheath tumor, osteosarcoma, myxochondrosarcoma, soft tissue sarcoma, oropharyngeal squamous cell carcinoma, chronic myeloid leukemia, epidermal squamous cell carcinoma, nasopharyngeal carcinoma, neuroblastoma, endometrial cancer, head and neck cancer, and cervical cancer.

[0060] In a preferred technical solution of the present disclosure, the cancer is metastatic cancer.

[0061] In a preferred technical solution of the present disclosure, the metastatic cancer is brain metastatic cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] Figure 1A-1B: Anti-tumor inhibition curves and inhibition rates of compound A, Divarasib, and their combination in the LU99 MTAP(- / -) deficient cell model

[0063] Figure 2A-2B: Anti-tumor inhibition curves and inhibition rates of compound C, Divarasib, and their combination in the LU99 MTAP(- / -) deficient cell model Specific embodiments

[0064] 1. Experimental compounds

[0065] The compound used in this study, Divarasib, was developed by Genentech USA and has the following chemical structure:

[0066] The chemical name of Compound A used in this test is (S)-4-amino-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide, which has the following chemical structure:

[0067] The chemical name of compound B used in this test is: 4-amino-N-cyclopropyl-7-fluoro-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide, which has the following chemical structure:

[0068] The chemical name of compound C used in this test is: (R)-4-amino-N-cyclopropyl-7-fluoro-N-(1-(5-trifluoromethyl)pyridin-2-yl)ethyl)imidazo[1,5-a]quinoxaline-8-carboxamide, which has the following chemical structure:

[0069] The chemical name of compound D used in this test is: (S)-4-amino-7-fluoro-N-methyl-N-(6-trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide, which has the following chemical structure:

[0070] The chemical name of compound E used in this test is: (S)-4-amino-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]pyrido[3,4-e]pyrazine-8-carboxamide, which has the following chemical structure:

[0071] The chemical name of compound F used in this test is: (S)-4-amino-N-(methyl-d3)-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide, which has the following chemical structure:

[0072] The chemical name of compound G used in this test is: (S)-4-amino-N-methyl-N-(6-(pentafluoro-λ6-sulfane)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline 8-carboxamide, which has the following chemical structure:

[0073] The chemical name of compound H used in this test is: (S)-4-amino-N-methyl-N-(6-(perfluoroethane)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide, which has the following chemical structure:

[0074] The chemical name of Compound I used in this test is: (S)-4-amino-7-fluoro-N-methyl-N-(6-trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide, which has the following chemical structure:

[0075] The chemical name of compound J used in this test is: (S)-4-amino-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide-1-d, which has the following chemical structure:

[0076] The chemical name of compound K used in this test is: (S)-4-amino-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide-3-d, which has the following chemical structure:

[0077] The chemical name of compound L used in this test is: (S)-4-amino-N-(methyl-d3)-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]pyrido[3,4-e]pyrazine-8-carboxamide, which has the following chemical structure:

[0078] The chemical name of the compound M used in this test is: (S)-4-amino-7-cyano-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide, which has the following chemical structure:

[0079] The chemical name of the compound N used in this test is: (R)-4-amino-N-methyl-d3-N-(1-(5-trifluoromethyl)pyridin-2-yl)ethyl)imidazo[1,5-a]quinoxaline-8-carboxamide, which has the following chemical structure:

[0080] The chemical name of Compound O used in this test is (R)-4-amino-N-methyl-N-(1-(5-(trifluoromethyl)pyridin-2-yl)ethyl)imidazo[1,5-a]quinoxaline-8-carboxamide, which has the following chemical structure:

[0081] Among them, the preparation of compound A, compound D, compound E, compound F, compound G, compound H, compound I, compound J, compound K, compound L and compound M refers to international patent application PCT / CN2023 / 120923;

[0082] Among them, the preparation of compound B, compound C, compound N and compound O refers to international patent application PCT / CN2023 / 111604, which is incorporated herein by reference in its entirety, as if they were fully cited herein.

[0083] 2. Combination of compounds in the NCI-H838 MTAP(- / -) deficient cell model

[0084] Cell lines and cell culture:

[0085] LU99 MTAP(- / -) cell line was purchased from Beijing Kangyuan Bochuang Biotechnology Co., Ltd. The cells were cultured in a cell culture medium containing 90% RPMI1640 and 10% fetal bovine serum at 37°C and 5% CO2. Experiments were performed only when the cells were in the logarithmic growth phase.

[0086] Combination drug testing:

[0087] Adjust the cell density using complete medium and then seed the cells in a 96-well cell culture plate. Culture the cells in the 96-well plate at 37°C and 5% CO2. Add the two test compounds to the cell plate using gradient dilutions. Continue culturing the cells in the 96-well plate at 37°C and 5% CO2 for 144 hours. Thaw the CTG reagent and equilibrate the cell plate to room temperature for 30 minutes. Add an equal volume of CTG solution to each well. Shake on an orbital shaker for 5 minutes to lyse the cells. Place the cell plate at room temperature for 20 minutes to stabilize the luminescence signal. Read the luminescence value and collect the data.

[0088] Data analysis: Cell inhibition rate (%) = 100-(Lum test drug-Lum culture medium control) / (Lum solvent control-Lum culture medium control) × 100%

[0089] Calcusyn was used to analyze the combination index (CI) values.

[0090] Example 1: Testing of single-drug compounds in the LU99 MTAP(- / -) deficient cell model

[0091] Table 1

[0092] Table 1 shows the IC values ​​of each compound in LU99 MTAP(- / -) deficient cells. 20 and IC 50 , and the maximum inhibition rate is given.

[0093] Example 2: Compound A combined with Divarasib in the LU99 MTAP(- / -) deficient cell model

[0094] The experimental results of the combination of Compound A and Divarasib on LU99 MTAP(- / -) deficient cells are shown in Table 2 for inhibition rate analysis, Figure 1A for inhibition curves, and Figure 1B for inhibition rates:

[0095] Table 2

[0096] FIG1B shows the inhibition rates of compound A (7.8 nM) and divarasib (0.7 nM) alone and in combination, indicating that the two compounds have a synergistic effect under these conditions.

[0097] The analysis of the CI combined index is shown in Table 3:

[0098] Table 3

[0099] Table 3 shows the combined effect index CI value of Compound A and Divarasib.

[0100] Example 3 Compound B and Divarasib combined in LU99 MTAP (- / -) deficient cell model

[0101] The experimental results of the combination of Compound B and Divarasib in the LU99 MTAP(- / -) deficient cell model are shown in Table 4 for inhibition analysis, Figure 2A for inhibition curves, and Figure 2B for inhibition rates.

[0102] Table 4

[0103] FIG2B shows the inhibition rates of Divarasib (0.74 nM) and Compound C (7.8 nM) alone and in combination, indicating that under this condition, the two compounds have an additive effect.

[0104] Analysis of CI combined index

[0105] Table 5

[0106] Table 5 shows the combined effect index CI value of compound C and Divarasib.

[0107] Conclusion: The combination of compound A and compound C with the CDK4 / 6 inhibitor Divarasib showed obvious synergistic-additive effects.

[0108] Although the preferred embodiment has been described above, it will be apparent to those skilled in the art that modifications may be made without departing from the invention. Such modifications are considered to be possible variations within the scope of the invention.

Claims

1. A pharmaceutical composition comprising a PRMT5 inhibitor having a first active substance and a KRAS inhibitor having a second active substance G12C Inhibitor compounds, wherein The PRMT5 inhibitor of the first active substance has the following structure: Wherein, in formula (I) or formula (II), R1 is selected from H, halogen, C1-C6 alkyl, halo(C1-C6 alkyl), CN; R2 is selected from H, C1-C6 alkyl, halo(C1-C6 alkyl), C3-C6 cycloalkyl; R3 represents H, halogen, C1-C6 alkyl, halo(C1-C6 alkyl), halo(C1-C6 alkoxy), or SF5; In formula II, R4 represents hydrogen or C1-C6 alkyl; X represents CR5 or N; Wherein, R5 represents hydrogen, halogen, C1-C6 alkyl, halo(C1-C6 alkyl), hydroxyl, -NH2, or CN.

2. The pharmaceutical composition according to claim 1, wherein In the structure of formula (I) or formula (II): R1 is selected from hydrogen, halogen, C1-C6 alkyl, halo(C1-C6 alkyl); R2 is selected from hydrogen, C1-C6 alkyl, halo(C1-C6 alkyl), C3-C6 cycloalkyl; R3 represents hydrogen, halogen, halo(C1-C6 alkyl), halo(C1-C6 alkoxy), or SF5; R4 represents hydrogen or methyl; In formula II, X represents CH or N.

3. The pharmaceutical composition according to claim 1, wherein R1 is selected from hydrogen or fluorine.

4. The pharmaceutical composition according to claim 1, wherein R2 is selected from cyclopropyl or methyl.

5. The pharmaceutical composition according to claim 1, wherein R3 represents CF3.

6. The pharmaceutical composition according to claim 1, wherein R4 represents hydrogen or methyl.

7. The pharmaceutical composition according to claim 1, wherein X represents N.

8. The pharmaceutical composition according to claim 1, wherein The PRMT5 inhibitor of the first active substance is selected from the following compounds or any combination thereof:

9. The pharmaceutical composition according to any one of claims 1 to 8, wherein the second active substance is KRAS G12C The inhibitor is selected from any one of the following compounds or their pharmaceutical salts: Sotorasib( and ), Divarasib (GDC-6036), Glecirasib (JAB-21822), LY3537982 (Eli Lilly and company), Garsorasib (D-1553), HBI-2438 (Huyabio International), Opnurasib (JDQ443), HS-10370 (Jiangsu Hansoh pharmaceutical company), IBI-351 (GFH925), BI-1823911 (Boehringer Ingelheim), JNJ-74699157 (ARS3248, Johnson & Johnson), RM-018 (Revolution Medicines), ARS-2102, RMC-4998, ASP2453, BI-2865, AZD4747, BI-0474, MK-108, BEBT-607, YL-15293, D3S-001, ABSK071, LY3499446, RMC-6291, SY-5933, GEC-255, RMC-4630 (SAR442720), BPI-421286, or any combination thereof.

10. The pharmaceutical composition according to claim 9, wherein the second active substance is KRAS G12C The inhibitor was selected from the Protac degrader LC2 (CAS: 2502156-03-6).

11. The pharmaceutical composition according to claim 9, wherein the second active substance is KRAS G12C Inhibitors selected from Sotorasib( and ), Opnurasib (JDQ443), LY3537982 (Eli Lilly and company), or a pharmaceutically acceptable salt thereof.

12. A method for treating cancer or tumors, comprising administering the pharmaceutical composition of any one of claims 1 to 11 to a subject in need thereof.

13. The method according to claim 12, wherein: The tumor or cancer is selected from the group consisting of glioblastoma multiforme, brain cancer, prostate cancer, pancreatic cancer, mantle cell lymphoma, non-Hodgkin lymphoma and diffuse large B-cell lymphoma, acute myeloid leukemia, acute lymphoblastic leukemia, multiple myeloma, non-small cell lung cancer, small cell lung cancer, breast cancer, triple-negative breast cancer, gastric cancer, colorectal cancer, ovarian cancer, bladder cancer, hepatocellular carcinoma, esophageal cancer, bile duct cancer, mesothelioma, laryngeal cancer, melanoma, malignant peripheral nerve sheath tumor, osteosarcoma, myxochondrosarcoma, soft tissue sarcoma, oropharyngeal squamous cell carcinoma, chronic myeloid leukemia, epidermal squamous cell carcinoma, nasopharyngeal carcinoma, neuroblastoma, endometrial cancer, head and neck cancer and cervical cancer.

14. The method according to claim 12, wherein: The cancer is a metastatic cancer.

15. The method according to claim 14, wherein The metastatic cancer is brain metastatic cancer.

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