Pharmaceutical composition containing PRMT5 inhibitor and mat2a inhibitor
Patent Information
- Application Number
- NZ835434
- Authority / Receiving Office
- NZ · NZ
- Patent Type
- Applications
- Current Assignee / Owner
- Priority Date
- 2024-03-04
- Filing Date
- 2025-01-24
- Publication Date
- 2025-07-31
AI Technical Summary
The existing PRMT5 inhibitors lack selectivity in MTAP-deleted cancer cell lines, resulting in a greater toxic effect on normal cells and cannot effectively target cancer cells accumulated by MTA.
Using a pharmaceutical composition containing a PRMT5 inhibitor and a MAT2A inhibitor, the MAT2A inhibitor is used to regulate the MTA level in cells, enhance the selective binding of the PRMT5 inhibitor, and improve the therapeutic effect on MTAP-deleted cancer cells through synergistic effects.
It significantly improves the selective inhibitory effect of MTAP-deleted cancer cells, reduces the toxicity to normal cells, and enhances the effectiveness and safety of cancer treatment.
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Abstract
Description
Pharmaceutical composition comprising a PRMT5 inhibitor and a MAT2A inhibitor Technical Field
[0001] The present invention belongs to the field of medicine, and specifically relates to a pharmaceutical composition for protecting a PRMT5 inhibitor and a MAT2A inhibitor. 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] Methionine adenosyltransferase 2A (MAT2A) is an enzyme that generates S-adenosylmethionine (SAM) from methionine (Met) and adenosine triphosphate (ATP). SAM is the primary methyl donor in cells for the methylation of a variety of substrates, including DNA, RNA, and proteins. One methyltransferase that utilizes SAM as a methyl donor is protein arginine N-methyltransferase 5 (PRMT5). Although PRMT5 activity requires SAM, PRMT5 is competitively inhibited by 5'-methylthioadenosine (MTA). Because MTA is part of the methionine salvage pathway, cellular MTA levels are kept low during the initiation of methylthioadenosine phosphorylase (MTAP).
[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 MAT2A inhibitor compound having a second active substance, 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 of the present disclosure, the MAT2A inhibitor of the second active substance is selected from any one of the following compounds or their pharmaceutical salts: IDE397, AG-270, S095033, ISM-3412 or any combination thereof.
[0028] In the preferred technical solution of the present disclosure, the MAT2A inhibitor of the second active substance is selected from IDE397, AG-270 or pharmaceutically acceptable salts thereof.
[0029] In the preferred technical solution of the present disclosure, the MAT2A inhibitor of the second active substance is selected from IDE397 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] In a preferred technical solution of the present disclosure, the cancer is metastatic cancer.
[0033] In a preferred technical solution of the present disclosure, the metastatic cancer is brain metastatic cancer.
[0034] 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.
[0035] 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".
[0036] 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.
[0037] 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.
[0038] 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.
[0039] "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.
[0040] "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.
[0041] As used herein, the term "cyano" refers to -CN.
[0042] 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.
[0043] As used herein, the terms "halo" and "halogen" refer to F, Cl, Br, or I.
[0044] 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.
[0045] 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.
[0046] 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.
[0047] 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.
[0048] "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.
[0049] 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."
[0050] 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.
[0051] 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.
[0052] 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.
[0053] 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.
[0054] 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.
[0055] 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.
[0056] 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.
[0057] 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.
[0058] 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.
[0059] The pharmaceutical composition may comprise a PRMT5 inhibitor and a MAT2A inhibitor for use in the methods of the present invention in a total amount of 0.01% to 99% by weight of the total composition, preferably 0.1% to 80% by weight of the total composition, and more preferably 0.1% to 50% by weight of the total composition. The weight ratio between the PRMT5 inhibitor and the MAT2A inhibitor may be in the range of 1:20 to 20:1, sometimes preferably 1:15 to 15:1, and sometimes more preferably 1:10 to 10:1.
[0060] 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.
[0061] 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. BRIEF DESCRIPTION OF THE DRAWINGS
[0062] Figure 1A-1B: Anti-tumor inhibition curves and inhibition rates of compound A, IDE397, and their combination in the NCI-H838 MTAP(- / -) deficient cell model
[0063] Figure 2A-2B: Anti-tumor inhibition curves and inhibition rates of compound B, IDE397, and their combination in the H838 MTAP(- / -) deficient cell model
[0064] Figure 3A-3B: Anti-tumor inhibition curves and inhibition rates of compound C, IDE397, and their combination in the H838 MTAP(- / -) deficient cell model
[0065] Figure 4A-4B: Anti-tumor inhibition curves and inhibition rates of compound D, IDE397, and their combination in the H838 MTAP(- / -) deficient cell model
[0066] Figure 5A-5B: Anti-tumor inhibition curves and inhibition rates of compound A, AG270, and their combination in the HCT116 MTAP(- / -) deficient cell model
[0067] Figure 6A-6B: Anti-tumor inhibition curves and inhibition rates of compound A, IDE397, and their combination in the HCT116 MTAP(- / -) deficient cell model Specific embodiments
[0068] 1. Experimental compounds
[0069] The compound IDE397 used in this study is compound A in IDEAYABIOSCIENCES patent WO2022 / 256806A1, which has the following chemical structure which are incorporated herein by reference in their entirety, as if fully incorporated herein.
[0070] The compound used in this study, AG-270, is Example 153 in Agios pharmaceuticals, Inc. patent WO2018 / 045071A1, which has the following chemical structure
[0071] 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:
[0072] 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:
[0073] 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:
[0074] 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:
[0075] 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:
[0076] 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:
[0077] 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:
[0078] 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:
[0079] 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:
[0080] 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:
[0081] 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:
[0082] 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:
[0083] 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:
[0084] 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:
[0085] 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:
[0086] 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;
[0087] 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.
[0088] 2. Combination of compounds in the NCI-H838 MTAP(- / -) deficient cell model
[0089] Cell lines and cell culture:
[0090] NCI-H838 MTAP(- / -)-deficient cells were purchased from Kangyuan Bochuang (China); cell culture medium, fetal bovine serum, and penicillin-streptomycin were purchased from Thermo Fisher Scientific (USA); and the Cell-Titer Glo assay kit was purchased from Promega (USA). Cells were cultured in 90% RPMI1640 medium containing 10% fetal bovine serum at 37°C and 5% CO2. Experiments were performed only when cells were in the logarithmic growth phase. Combination drug testing:
[0091] 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.
[0092] Data Analysis:
[0093] Cell inhibition rate (%) = 100-(Lum test drug-Lum culture medium control) / (Lum solvent control-Lum culture medium control) × 100%
[0094] Calcusyn was used to analyze the combination index (CI) values.
[0095] Example 1: Single-drug compound testing in the NCI-H838 MTAP(- / -) deficient cell model
[0096] Table 1
[0097] Table 1 shows the IC values of each compound in NCI-H838 MTAP(- / -) deficient cells. 20 and IC 50 , and the maximum inhibition rate is given.
[0098] Example 2: Compound A and IDE397 combined in NCI-H838 MTAP(- / -) deficient cell model
[0099] The experimental results of the combination of compound A and IDE397 on H838 are shown in Table 2 for inhibition rate analysis, Figure 1A for inhibition curves, and Figure 1B for inhibition rates:
[0100] Table 2
[0101] FIG1B shows the inhibition rates of compound A (37 nM) and IDE397 (12 nM) alone and in combination, indicating that under these conditions, the two compounds have a very strong synergistic effect.
[0102] The analysis of the CI combined index is shown in Table 3:
[0103] Table 3
[0104] Table 3 shows the combined effect index CI value of compound A and IDE397.
[0105] Example 3 Compound B and IDE397 combined in NCI-H838 MTAP(- / -) deficient cell model
[0106] The experimental results of the combination of compound B and IDE397 in the NCI-H838 MTAP(- / -) deficient cell model are shown in Table 4 for inhibition analysis, Figure 2A for inhibition curves, and Figure 2B for inhibition rates.
[0107] Table 4
[0108] FIG2B shows the inhibition rates of compound B (37 nM) and IDE397 (12 nM) alone and in combination, indicating that under this condition, the two compounds have a strong synergistic effect.
[0109] Analysis of CI combined index
[0110] Table 5
[0111] Table 5 shows the combined effect index CI value of compound B and IDE397.
[0112] Example 4: Combination of Compound C and IDE397 in the NCI-H838 MTAP(- / -) Deficient Cell Model
[0113] The experimental results of the combination of compound C and IDE397 on NCI-H838 MTAP(- / -) deficient cells are shown in Table 6 for inhibition analysis, Figure 3A for inhibition curves, and Figure 3B for inhibition rates.
[0114] Table 6
[0115] FIG3B shows the inhibition rates of compound C (37 nM) and IDE397 (12 nM) alone and in combination, indicating that under this condition, the two compounds have a strong synergistic effect.
[0116] The analysis of the CI combined index is shown in Table 7
[0117] Table 7
[0118] Table 7 above shows the combined effect index CI value of compound C and IDE397.
[0119] Example 5: Compound D and IDE397 combined in NCI-H838 MTAP(- / -) deficient cell model
[0120] The experimental results of the combination of compound D and IDE397 on NCI-H838 MTAP(- / -) deficient cells are shown in Table 8 for inhibition analysis, Figure 4A for inhibition curves, and Figure 4B for inhibition rates:
[0121] Table 8
[0122] FIG4B shows the inhibition rates of compound D (37 nM) and IDE397 (12 nM) alone and in combination, indicating that under this condition, the two compounds have a strong synergistic effect.
[0123] The analysis of the CI combined index is shown in Table 9:
[0124] Table 9
[0125] Table 9 shows the combined effect index CI value of compound D and IDE397.
[0126] 3. Combination of compounds in HCT116 MTAP(- / -) deficient cells
[0127] Cell lines and cell culture:
[0128] HCT116 MTAP(- / -)-deficient cells were provided by Beijing Kangyuan Bochuang Company. 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.
[0129] Combination drug testing:
[0130] Adjust the cell density with complete medium and then seed the cells in a 384-well cell culture plate. Culture the cells in the 384-well plate at 37°C and 5% CO2. Add the two test compounds to the cell plate using a gradient dilution. Continue culturing the cells in the 384-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.
[0131] Data Analysis
[0132] Cell inhibition rate (%) = 100-(Lum test drug-Lum culture medium control) / (Lum solvent control-Lum culture medium control) × 100%
[0133] Calcusyn was used to analyze the combination index (CI) values.
[0134] Example 6: The results of single-drug testing of the compound on HCT116 MTAP(- / -) deficient cells are shown in Table 10
[0135] Table (10)
[0136] Table 10 shows the IC values of each compound on HCT116 MTAP(- / -) deficient cells. 20 and IC 50 , and the maximum inhibition rate is given.
[0137] Example 7: Analysis of the inhibitory rate of the combined use of compound A and AG270 on HCT116 MTAP(- / -) deficient cells:
[0138] The results of the combined use of compound A and AG270 on HCT116 MTAP(- / -) deficient cells are shown in Table 11 for inhibition analysis, Figure 5A for inhibition curves, and Figure 5B for inhibition rates:
[0139] Table 11
[0140] FIG5B shows the inhibition rates of compound A (4.1 nM) and AG270 (12.3 nM) alone and in combination, indicating that under this condition, the two compounds have a certain synergistic effect.
[0141] The analysis of the CI combined index is shown in Table 12:
[0142] Table 12
[0143] Conclusion: The combined use of compound A and AG270 on HCT116 MTAP(- / -) deficient cells showed a CI value ranging from 0.2 to 2.7, indicating a synergistic to additive effect.
[0144] Example 8: Analysis of the inhibitory rate of the combined use of compound A and IDE397 on HCT116 MTAP(- / -) deficient cells:
[0145] The experimental results of the combination of Compound A and AIDE397 on HCT116 MTAP(- / -) deficient cells are shown in Table 13 for inhibition analysis, Figure 6A for inhibition curves, and Figure 6B for inhibition rates:
[0146] Table 13
[0147] FIG6B shows the inhibition rates of compound A (4.1 nM) and IDE397 (4.1 nM) alone and in combination, indicating that under this condition, the two compounds have a certain synergistic effect.
[0148] The analysis of the CI combined index is shown in Table 14:
[0149] Table 14
[0150] Conclusion: The CI values of compound A and IDE397 in HCT116 MTAP(- / -) deficient cells ranged from 0.2 to 2.9, indicating that the two compounds had a synergistic effect.
[0151] Summary: Comparing the combined effects of compound A with IDE397 and AG270, it was concluded that compound A and IDE397 had a stronger synergistic effect.
Claims
1. A pharmaceutical composition comprising a PRMT5 inhibitor having a first active substance and a MAT2A inhibitor compound having a second active substance, wherein, The PRMT5 inhibitor of the first active substance described has the following structural formula (I) or formula (II): Among them, 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), 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), hydroxy, -NH2, 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), SF5; R4 represents hydrogen, 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 inhibitors of the first active substance are selected from the following compounds or any combination thereof:
9. The pharmaceutical composition according to any one of claims 1-8, wherein the MAT2A inhibitor of the second active substance is selected from any one of the following compounds or its pharmaceutically acceptable salts: IDE397, AG-270, S095033, ISM-3412 or any combination thereof.
10. The pharmaceutical composition according to claim 9, wherein the MAT2A inhibitor of the second active substance is selected from IDE397, AG-270 or its pharmaceutically acceptable salts.
11. The pharmaceutical composition according to claim 9, wherein the MAT2A inhibitor of the second active substance is selected from IDE397 or its pharmaceutically acceptable salts.
12. A method for treating cancer or tumor, comprising administering the pharmaceutical composition according to any one of claims 1-11 to an individual in need.
13. The method according to claim 12, wherein, The tumor or cancer is selected from: 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, cholangiocarcinoma, 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 metastatic cancer.
15. The method according to claim 14, wherein, The metastatic cancer is brain metastatic cancer.