Combination therapy comprising MAT2A inhibitor and topoisomerase inhibitor, splice inhibitor sulfonamide or KIF18 inhibitor

Abstract

Provided herein are combinations of a methionine adenylyltransferase II alpha (MAT2A) inhibitor and a topoisomerase inhibitor, splicing inhibitor sulfonamide, or a KIF18 inhibitor. Also provided are methods of using such combinations to treat diseases or conditions, such as cancer.

Description

[0001] Cross-references to related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 495,911, filed April 13, 2023, and U.S. Provisional Application No. 63 / 598,799, filed November 14, 2023, the entire contents of which are incorporated herein by reference and for all purposes. Background Technology

[0003] Cancer is a leading cause of death worldwide. The limitation of mainstream treatments (such as chemotherapy and immunotherapy) is that their cytotoxic effects are not limited to cancer cells but can also cause adverse side effects in normal tissues.

[0004] Methionine adenosine transferase 2A (MAT2A) is an enzyme that uses methionine (Met) and adenosine triphosphate (ATP) to generate s-adenosine methionine (SAM). SAM is a major methyl donor in cells used for methylation of various substrates, including DNA, RNA, and proteins. One methyltransferase that uses SAM as a methyl donor is protein arginine N-methyltransferase 5 (PRMT5). Although SAM is essential for PRMT5 activity, PRMT5 is competitively inhibited by 5'-methionine (MTA). Because MTA is part of the methionine rescue pathway, cellular MTA levels remain low during the initiation of methionine phosphorylase (MTAP).

[0005] MTAP is located at a locus on chromosome 9, which is typically absent in cells from patients with cancers of several originating tissues, including the central nervous system, pancreas, esophagus, bladder, and lung (cBioPortal database). MTAP deficiency leads to the accumulation of MTA, and MTAP-deficient cells are more dependent on SAM production, and thus dependent on MAT2A activity, compared to MTAP-expressing cells. In a shRNA cell line screening of approximately 400 cancer cell lines, MAT2A knockdown resulted in a greater proportion of MTAP-deficient cells losing viability compared to MTAP-expressing WT cells (see McDonald et al., 2017). Cell 170, 577-592). Furthermore, inducible knockdown of the MAT2A protein reduces tumor growth in vivo (see Marjon et al., 2016). Cell Reports 15(3), 574-587). These results suggest that MAT2A inhibitors may offer a novel treatment option for cancer patients, including those with MTAP-deficient tumors.

[0006] DNA topoisomerases are enzymes found in the cell nucleus that catalyze the breaking and rejoining of DNA strands, thereby controlling the topological state of DNA. Recent studies have also shown that topoisomerases are involved in regulating template supercoiling during RNA transcription. There are two main types of mammalian topoisomerases. DNA topoisomerase I catalyzes changes in the topological state of double-stranded DNA through a transient single-strand break-bind cycle. Conversely, mammalian topoisomerase II alters the topological structure of DNA by inducing a transient double-strand break in the enzyme bridge, followed by the strand passing through and resealing. The antitumor activity associated with topoisomerase toxins is related to their ability to stabilize enzyme-DNA cleavable complexes. This drug-induced stabilization of the enzyme-DNA cleavable complex effectively converts the enzyme into a cytotoxic agent.

[0007] Indisulam is an aryl sulfonamide drug with selective anticancer activity. Its mechanism of action and the basis of its selectivity are not fully understood. Many cancer cell lines from hematopoietic and lymphoid lineages are sensitive to indisulam, and this sensitivity is correlated with DCAF15 expression levels. Two other clinically tested sulfonamides, tasisulam and CQS, have the same mechanism of action as indisulam. These drugs are often referred to as SPLAMs (splicing inhibitor sulfonamides).

[0008] Kinesins are molecular motors that play crucial roles in cell division and the transport of intracellular vesicles and organelles. Mitotic kinesins function in multiple ways, including spindle assembly, chromosome segregation, centromere separation, and dynamics (reviewed in O. Rath and F. Kozielski, Nature Review Cancer, 12:527-39, 2012). Based on sequence homology within their motor domains, human kinesins are divided into 14 subfamilies. The ATPase activity of this domain drives unidirectional movement along microtubules (MTs). The non-motor domains of these proteins are responsible for cargo attachment. This cargo can include a variety of membrane-bound organelles, signal transduction scaffold systems, and chromosomes. Kinesins use the energy from ATP hydrolysis to move their cargo along polarized microtubules. Therefore, kinesins are often referred to as "positive-terminal" or "negative-terminal" directional motors.

[0009] The KIF18A gene belongs to the kinesin-8 subfamily and is a positive-terminal orientation motor. KIF18A is thought to affect the dynamics of the positive ends of kinetochore microtubules, thereby controlling proper chromosome positioning and spindle tension. Depletion of human KIF18A leads to spindle elongation, increased metaphase chromosome oscillations, and activation of mitotic spindle assembly checkpoints in HeLa cervical cancer cells (MI Mayr et al., Current Biology 17, 488-98, 2007). KIF18A is overexpressed in various types of cancer, including but not limited to colon cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head cancer, cervical cancer, cervical cancer, and ovarian cancer. Furthermore, gene deletion, knockdown, or inhibition of KIF18A affects the mitotic spindle in cancer cell lines. Specifically, inhibition of KIF18A has been found to induce mitotic cell arrest, a known vulnerability that can promote cell death in mitosis through apoptosis, mitotic catastrophe or multipolar-driven lethality, or death following interphase slippage. Therefore, people have developed a strong interest in finding inhibitors of the KIF18A protein.

[0010] Despite recent advances in cancer treatment, more effective and / or enhanced treatments are still needed for individuals affected by cancer. Summary of the Invention

[0011] This article provides a combination product comprising a methionine adenosyltransferase IIα (MAT2A) inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor. The combination product can be used to treat a variety of cancers, including solid tumors. The combination product can also be used to treat a variety of diseases or conditions that can be treated by inhibiting MAT2A. The combination product can also be used to treat MTAP-deficient tumors.

[0012] In one implementation, this document provides a combination of a MAT2A inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor.

[0013] In one embodiment, this document provides a pharmaceutical composition comprising a therapeutically effective amount of a methionine adenosyltransferase IIα (MAT2A) inhibitor, and a second pharmaceutical composition comprising a therapeutically effective amount of a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor. In some embodiments, the second pharmaceutical composition comprises a topoisomerase inhibitor. In some embodiments, the second pharmaceutical composition comprises SPLAM. In some embodiments, the second pharmaceutical composition comprises a KIF18 inhibitor.

[0014] In one embodiment, this document provides a method for treating cancer in a subject in need, the method comprising administering to the subject a combination comprising a MAT2A inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor, thereby treating the subject's cancer.

[0015] In one embodiment, this document provides a method for treating cancer in a subject in need, the method comprising administering to the subject a combination comprising a MAT2A inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM) or a KIF18 inhibitor, and at least one pharmaceutically acceptable carrier, thereby treating the subject's cancer.

[0016] In yet another implementation, the cancer is characterized by reduced or absent expression of the methionine phosphorylase (MTAP) gene, absence of the MTAP gene, reduced function of the MTAP protein, reduced or absent levels of the MTAP protein, accumulation of MTA, or a combination thereof.

[0017] In one embodiment, this document provides a method for treating cancer in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a MAT2A inhibitor and a therapeutically effective amount of a pharmaceutical composition comprising a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor, thereby treating the subject's cancer.

[0018] In one embodiment, this document provides a method for treating a disease or condition treatable by inhibiting MAT2A in a subject of need, the method comprising administering to the subject a combination comprising a methionine adenosyltransferase IIα (MAT2A) inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor, thereby treating the subject's disease or condition. In one embodiment, the disease or condition is cancer.

[0019] In one embodiment, this document provides a method for treating a disease or condition treatable by inhibiting MAT2A in a subject of need, the method comprising administering to the subject a combination comprising a MAT2A inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM) or a KIF18 inhibitor, and at least one pharmaceutically acceptable carrier, thereby treating the subject's disease or condition. In one embodiment, the disease or condition is cancer.

[0020] In one embodiment, the MAT2A inhibitor is a compound of formula (I):

[0021] (I)

[0022] Or a pharmaceutically acceptable salt thereof, wherein the variables of formula (I) are defined below.

[0023] In another embodiment, the MAT2A inhibitor is compound A having the following structural formula:

[0024] Compound A

[0025] Or its pharmaceutically acceptable salt.

[0026] In another embodiment, the MAT2A inhibitor is compound A1 having the following structural formula:

[0027] Compound A1

[0028] Or its pharmaceutically acceptable salt.

[0029] MAT2A inhibitors used in the combination therapies described herein are described in WO 2020 / 123395 (PCT / US19 / 65260). Common and specific compounds described in the patent applications are incorporated herein by reference and may be used to treat the cancers described herein.

[0030] In one implementation, the KIF18 inhibitor is compound B:

[0031] Compound B

[0032] Or a pharmaceutically acceptable salt thereof. The chemical name of compound B is N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxyethyl)sulfonamido)-2-(6-azaspiro[2.5]oct-6-yl)benzamide. Compound B is also known as "sovilnesib" or "AMG650". Compound B also includes its pharmaceutically acceptable salt.

[0033] In another implementation, SPLAM stands for Indisuran:

[0034] Indisuran

[0035] Or its pharmaceutically acceptable salt. The chemical name of Indisulan is N-(3-chloro-1H-indol-7-yl)benzene-1,4-disulfonamide. Indisulan also includes its pharmaceutically acceptable salt.

[0036] In another implementation, SPLAM is E7820:

[0037] E7820

[0038] Or its pharmaceutically acceptable salts. The chemical name of E7820 is 3-cyano-N-(3-cyano-4-methyl-1H-indol-7-yl)benzenesulfonamide. E7820 also includes its pharmaceutically acceptable salts.

[0039] In another embodiment, SPLAM is chloroquinoxaline sulfonamide:

[0040] Chloroquinoxaline sulfonamide

[0041] Or its pharmaceutically acceptable salts. The chemical name of chloroquinoxaline sulfonamide is 4-amino-N-(5-chloroquinoxal-2-yl)benzenesulfonamide, also known as "CQS". Chloroquinoxaline sulfonamide also includes its pharmaceutically acceptable salts.

[0042] In another implementation, SPLAM stands for Tassisuland:

[0043] Tassisuran

[0044] Or its pharmaceutically acceptable salts. The chemical name of tassisulan is N-((5-bromothiophen-2-yl)sulfonyl)-2,4-dichlorobenzamide. Tassisulan also includes its pharmaceutically acceptable salts.

[0045] In one implementation, the topoisomerase inhibitor is a type I topoisomerase inhibitor.

[0046] In another embodiment, the type I topoisomerase inhibitor is 10-hydroxycamptothecin:

[0047] 10-Hydroxycamptothecin

[0048] Or its pharmaceutically acceptable salts. The chemical name of 10-hydroxycamptothecin is (S)-4-ethyl-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indolazino[1,2-b]quinoline-3,14(4H)-dione. 10-hydroxycamptothecin also includes its pharmaceutically acceptable salts.

[0049] In another embodiment, the type I topoisomerase inhibitor is irinotecan:

[0050] Irinotecan

[0051] Or its pharmaceutically acceptable salts. Irinotecan's chemical name is (S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolazino[1,2-b]quinoline-9-yl[1,4'-bipiperidine]-1'-carboxylate. Irinotecan also includes its pharmaceutically acceptable salts.

[0052] In another embodiment, the type I topoisomerase inhibitor is topotecan:

[0053] Topotecan

[0054] Or its pharmaceutically acceptable salts. The chemical name of topotecan is (S)-10-((dimethylamino)methyl)-4-ethyl-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indolazino[1,2-b]quinoline-3,14(4H)-dione. Topotecan also includes its pharmaceutically acceptable salts.

[0055] In one implementation, the topoisomerase inhibitor is a type II topoisomerase inhibitor.

[0056] In another embodiment, the type II topoisomerase inhibitor is daunorubicin:

[0057] daunorubicin

[0058] Or its pharmaceutically acceptable salts. The chemical name of daunorubicin is (8S,10S)-8-acetyl-10-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,8,11-trihydroxy-1-methoxy-7,8,9,10-tetrahydrotetrabenzo-5,12-dione. Daunorubicin also includes its pharmaceutically acceptable salts.

[0059] In another embodiment, the type II topoisomerase inhibitor is doxorubicin:

[0060] Doxorubicin

[0061] Or its pharmaceutically acceptable salts. The chemical name of doxorubicin is (8S,10S)-10-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,8,11-trihydroxy-8-(2-hydroxyacetyl)-1-methoxy-7,8,9,10-tetrahydrotetraphenyl-5,12-dione. Doxorubicin also includes its pharmaceutically acceptable salts.

[0062] In another embodiment, the type I topoisomerase inhibitor is etoposide:

[0063] Etoposide

[0064] Or its pharmaceutically acceptable salts. The chemical name of etoposide is (5R,5aR,8aR,9S)-9-(((2R,6R,7R,8R,8aS)-7,8-dihydroxy-2-methylhexahydropyrano[3,2-d][1,3]dioxin-6-yl)oxy)-5-(4-hydroxy-3,5-dimethoxyphenyl)-5,8,8a,9-tetrahydrofurano[3',4':6,7]naphtho[2,3-d][1,3]dioxane-6(5aH)-one. Etoposide also includes its pharmaceutically acceptable salts. Attached Figure Description

[0065] Figures 1A-1 to 1A-5 and Figures 1B-1 to 1B-2 Compound B (sowecoxib) was demonstrated as a single agent in five MTAP-deficient cell lines. Figures 1A-1 to 1A-5 ) and two wild-type cell lines ( Figure 1B-1 and 1B-2 Growth inhibition in ).

[0066] Figures 2A-1 to 2G Compound A and 10-hydroxycamptothecin were shown in thirteen (13) species. MTAP Growth inhibition in defective cell lines.

[0067] Figures 3A to 3G-2 Compound A and 10-hydroxycamptothecin were shown in thirteen (13) species. MTAP Loewe synergistic effect in defective cell lines.

[0068] Figures 4A-1 to 4G Compound A and irinotecan were shown in thirteen (13) species. MTAP Growth inhibition in defective cell lines.

[0069] Figures 5A to 5G-2 Compound A and irinotecan were shown in thirteen (13) species. MTAPLoewe synergistic effect in defective cell lines.

[0070] Figures 6A-1 to 6G Compound A and topotecan were shown in thirteen (13) species. MTAP Growth inhibition in defective cell lines.

[0071] Figures 7A to 7G-2 Compound A and topotecan were shown in thirteen (13) species. MTAP Loewe synergistic effect in defective cell lines.

[0072] Figures 8A-1 to 8G Compound A and daunorubicin were shown in thirteen (13) species. MTAP Growth inhibition in defective cell lines.

[0073] Figures 9A to 9G-2 Compound A and daunorubicin were shown in thirteen (13) species. MTAP Loewe synergistic effect in defective cell lines.

[0074] Figures 10A-1 to 10G Compound A and doxorubicin were shown in thirteen (13) species. MTAP Growth inhibition in defective cell lines.

[0075] Figures 11A to 11G-2 Compound A and doxorubicin were shown in thirteen (13) species. MTAP Loewe synergistic effect in defective cell lines.

[0076] Figures 12A-1 to 12G Compound A and etoposide were shown in thirteen (13) species. MTAP Growth inhibition in defective cell lines.

[0077] Figures 13A to 13G-2 Compound A and etoposide were shown in thirteen (13) species. MTAP Loewe synergistic effect in defective cell lines.

[0078] Figure 14 The antitumor activity of compound A and irinotecan, alone or in combination, was demonstrated in the RT-112 / 84 bladder CDX model.

[0079] Figure 15 The changes in individual tumor volume after administration of irinotecan alone or in combination of compound A and irinotecan in the RT-112 / 84 CDX model are shown.

[0080] Figure 16 The antitumor activity of compound A and irinotecan in the gastric CDX model MKN45 was demonstrated.

[0081] Figure 17The antitumor activity of compound A and irinotecan in the gastric CDX model LMSU was demonstrated. Detailed Implementation

[0082] This article provides a combination therapy comprising a methionine adenosyltransferase IIα (MAT2A) inhibitor or a pharmaceutically acceptable salt thereof, and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof. This combination therapy can be used to treat a variety of cancers. In another aspect, this combination therapy can be used to treat any of MAT2A-related diseases. In yet another aspect, this combination therapy can be used to treat diseases or conditions that can be treated by inhibiting MAT2A.

[0083] definition

[0084] The definitions of the various terms used herein are listed below. Unless otherwise limited individually in particular cases or as part of a larger group, these definitions apply to the terms used in this specification and claims.

[0085] Unless otherwise defined, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art. Typically, the nomenclature used herein, as well as laboratory procedures in cell culture, molecular genetics, organic chemistry, and peptide chemistry, are those well-known and commonly used in the art.

[0086] As used herein, the articles “a” and “an” refer to one (or a kind) or more than one (or a kind) (i.e., at least one (or a kind)) grammatical object. For example, “an element” means one element or more elements. Furthermore, the use of the term “including” and other forms such as “include,” “includes,” and “included” is not restrictive.

[0087] As used herein, the term “about” will be understood by those skilled in the art and will vary to some extent depending on the context of its use. As used herein, when referring to measurable values ​​(such as quantity, duration, etc.), the term “about” is intended to cover variations of ±20% or ±10% relative to a specified value, including ±5%, ±1%, and ±0.1%, as such variations are suitable for performing the disclosed methods.

[0088] As used in the specification and claims, the term "comprising" may include embodiments "consisting of" and "substantially consisting of". As used herein, the terms "comprising", "including", "having", "having", "may", "containing", and variations thereof are intended as open transitional phrases, terms, or words that require the presence of a specified ingredient / step and allow the presence of other ingredients / steps.

[0089] It should be noted that ratios, concentrations, amounts, and other numerical data may be expressed in range format herein. It should be understood that such range format is used for convenience and brevity, and therefore should be interpreted flexibly as including not only the numerical values ​​explicitly listed as the boundaries of the range, but also all individual numerical values ​​or subranges contained within that range, as if each numerical value and subrange were explicitly listed. Furthermore, the phrase "about 'x' to 'y'" includes "about 'x' to about 'y'".

[0090] As used herein, the terms “combination,” “therapeutic combination,” “drug combination,” or “combination product” refer to a fixed combination in the form of a single dose unit, or a non-fixed combination in the form of a single dose, or a kit for combination administration, wherein two or more therapeutic agents may be administered simultaneously independently or separately at time intervals.

[0091] The term "combination therapy" refers to the administration of two or more therapeutic agents to treat the therapeutic symptoms or conditions described in this disclosure. The administration encompasses the combined administration of these therapeutic agents in a substantially simultaneous manner, such as in a single formulation having a fixed proportion of the active ingredients or in a separate formulation of each active ingredient (e.g., capsules and / or intravenous formulations). Furthermore, the administration also encompasses the use of each type of therapeutic agent sequentially or individually at approximately simultaneous or at different times. Regardless of whether the active ingredient is administered as a single formulation or in separate formulations, the drugs are administered to the same patient as part of the same course of treatment. In any case, the treatment regimen will provide a beneficial effect, such as a synergistic effect, in the treatment of the disease or condition described herein.

[0092] As used herein, the term “treating / treatment” refers to suppressing disease; for example, suppressing the disease, illness, or symptom of an individual who is experiencing or exhibiting the pathology or symptomology of a disease, illness, or symptom (i.e., preventing further development of the pathology and / or symptomology), or improving disease; for example, improving the disease, illness, or symptom of an individual who is experiencing or exhibiting the pathology or symptomology of a disease, illness, or symptom (i.e., reversing the pathology and / or symptomology), such as reducing the severity of the disease.

[0093] As used herein, the term "prevent" means the absence of the development of a symptom or disease (if the development of said symptom or disease has not yet occurred), or the absence of further development of the symptom or disease (if said symptom or disease has already developed). The ability of the combination therapies presented herein to prevent some or all of the symptoms associated with the symptom or disease is also considered.

[0094] As used herein, the terms “patient,” “individual,” or “subject” refer to a human or a non-human mammal. Non-human mammals include, for example, livestock and pets, such as sheep, cattle, pigs, dogs, cats, and marine mammals. Preferably, the patient, subject, or individual is a human.

[0095] As used herein, the terms "effective amount," "pharmaceutical effective amount," and "therapeutic effective amount" refer to an amount of an agent that is non-toxic but sufficient to provide the desired biological outcome. This outcome may be the reduction or alleviation of signs, symptoms, or causes of disease, or any other desired alteration of a biological system. The appropriate therapeutic amount in any individual case can be determined by a person skilled in the art using routine laboratory methods.

[0096] As used herein, the term “pharmaceutically acceptable” means that a material (such as a carrier or diluent) does not eliminate the biological activity or properties of a compound and is relatively non-toxic, i.e., can be administered to an individual without causing undesirable biological effects or interacting in a harmful manner with any of the components of a composition containing it.

[0097] As used herein, the term "pharmaceutically acceptable salt" refers to a derivative of a disclosed compound in which the parent compound is modified by converting an existing acidic or basic moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic acid or organic acid salts having basic residues such as amines; alkali metal salts or organic salts having acidic residues such as carboxylic acids; etc. Pharmaceutically acceptable salts described herein include, for example, conventional non-toxic salts of parent compounds formed from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts discussed herein can be synthesized by conventional chemical methods from parent compounds containing basic or acidic moieties. Typically, such salts are prepared by reacting the free acidic or basic form of these compounds with a stoichiometric amount of a suitable base or acid in water, in an organic solvent, or in a mixture of both; non-aqueous media such as diethyl ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are typically used. The phrase "pharmaceutically acceptable salt" is not limited to a single salt or a 1:1 salt. For example, "pharmaceutically acceptable salt" also includes disalts, such as dihydrochlorides. A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th edition, Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is incorporated herein by reference in its entirety.

[0098] As used herein, the term "composition" or "pharmaceutical composition" refers to a mixture of at least one compound with a pharmaceutically acceptable carrier. The pharmaceutical composition facilitates administration to a patient or subject. Various techniques for administering compounds exist in the art, including but not limited to intravenous, oral, aerosol, parenteral, ocular, pulmonary, and topical administration.

[0099] As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or carrier, such as a liquid or solid filler, stabilizer, dispersant, suspending agent, diluent, excipient, thickener, solvent, or encapsulating material, that is involved in carrying or transporting a compound useful to a patient and enabling the compound to perform its intended function. Typically, such constructs are delivered or transported from one organ or body part to another. Each carrier must be "acceptable," meaning it is compatible with other components of the formulation, including the compounds disclosed herein, and is harmless to the patient. Examples of materials that can serve as pharmaceutically acceptable carriers include: sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; powdered tragacanth gum; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerol, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; surfactants; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol; phosphate buffer solutions; and other non-toxic compatible substances used in pharmaceutical preparations.

[0100] As used herein, "pharmaceutically acceptable carrier" also includes any and all coatings, antimicrobial and antifungal agents, and absorption delayers that are compatible with the activity of the compounds disclosed herein and are physiologically acceptable to patients. Complementary active compounds may also be incorporated into the composition. Other additional ingredients that may be included in the pharmaceutical composition are known in the art and described, for example, in Remington's Pharmaceutical Sciences (Genaro, Ed., MackPublishing Co., 1985, Easton, PA), references which are incorporated herein by reference.

[0101] As used herein, the term "single formulation" refers to a single carrier or mediator formulated to deliver a therapeutically effective amount of two therapeutic agents to a patient. A single mediator is designed to deliver a therapeutically effective amount of each agent together with any pharmaceutically acceptable carrier or excipient. In some embodiments, the mediator is a tablet, capsule, pill, or patch. In other embodiments, the mediator is a solution or suspension.

[0102] As used in this article, "methionine adenosyltransferase II α inhibitor" or "MAT2A inhibitor" refers to an agent that regulates MAT2A activity or inhibits the production of S-adenosylmethionine (SAM) by methionine adenosyltransferase 2A (MAT2A).

[0103] The combinations of agents described herein can exhibit synergistic effects. As used herein, the term "synergistic effect" or "synergy" refers to the effect produced by two agents (e.g., MAT2A inhibitors and topoisomerase inhibitors, splicing inhibitors sulfonamides (SPLAM), or KIF18 inhibitors) that, for example, slow the symptomatic progression of cancer or its symptoms, is greater than the simple additive effect of each agent when administered alone. Synergistic effects can be calculated using suitable methods, such as the Sigmoid-Emax equation (Holford, NHG and Scheiner, LB, Clin. Pharmacokinet. 6:429-453 (1981)), the Loewe additive equation (Loewe, S. and Muischnek, H., Arch. Exp. PatholPharmacol. 114: 313-326 (1926)), and the intermediate-effect equation (Chou, TC and Talalay, P., Adv. Enzyme Regul. 22: 27-55 (1984)). These equations can be applied to experimental data to generate corresponding graphs to help assess the effects of drug combinations. The corresponding graphs associated with the equations mentioned above are concentration-response curves, equivalence curves, and combination exponential curves.

[0104] In one embodiment, this document provides a combination therapy comprising a therapeutically effective amount of a MAT2A inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor. The “therapeuticly effective amount” of the combination of agents (i.e., the combination of a MAT2A inhibitor with a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor) is an amount sufficient to provide an observable improvement in baseline clinically observable signs and symptoms relative to the condition treated with said combination.

[0105] "Alkyl" refers to a straight-chain saturated monovalent hydrocarbon group having one to six carbon atoms (i.e., C1-6 means one to six carbon atoms) or a branched saturated monovalent hydrocarbon group having three to six carbon atoms (i.e., C3-6 means three to six carbon atoms). Alkyl groups can include any number of carbon atoms, such as C1-C6. 1-2 C 1-3 C 1-4 C 1-5 C 1-6 C 2-3 C 2-4 C 2-5 C 2-6 C 3-4 C 3-5 C 3-6C 4-5 C 4-6 and C 5-6 Examples of alkyl groups include methyl, ethyl, propyl, 2-propyl, butyl, pentyl, etc. Those skilled in the art will recognize that the term "alkyl" may include "alkylene" groups.

[0106] "Amino" refers to -NH2.

[0107] "Cycloalkyl" refers to a monocyclic monovalent hydrocarbon group with three to six carbon atoms (e.g., C64). 3-6 Cycloalkyl groups (which may be saturated or contain a double bond) can include any number of carbons, such as C60, C70, C80, C9 ... 3-6 C 4-6 and C 5-6 Partially unsaturated cycloalkyl groups have one or more bis(alkyl groups) in the ring, but the cycloalkyl group is not aromatic. Saturated monocyclic cycloalkyl rings include, for example, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0108] "Halogen" refers to fluorine, chlorine, bromine or iodine, with fluorine or chlorine being preferred.

[0109] "Halogenated alkyl" means an alkyl group as defined above that is substituted with one to five halogen atoms (such as fluorine or chlorine), including groups substituted with different halogens, such as -CH2Cl, -CF3, -CHF2, -CH2CF3, -CF2CF3, -CF(CH3)2, etc. When an alkyl group is substituted only with fluorine, it may be referred to as a fluoroalkyl group in this application. As for alkyl groups, halogenated alkyl groups may have any suitable number of carbon atoms, such as C 1-6 .

[0110] Combination products

[0111] This document provides a combination product comprising a MAT2A inhibitor or a pharmaceutically acceptable salt thereof; and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof. This combination product is also referred to herein as a combination therapy. The combination product can be used to treat a variety of cancers, including solid tumors. In another aspect, the combination product can be used to treat any of MAT2A-related diseases. In another aspect, the combination product can be used to treat diseases or conditions that can be treated by inhibiting MAT2A. In another aspect, the combination product can be used to treat MTAP-deficient tumors.

[0112] In one embodiment, this document provides a combination of a MAT2A inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor. In some embodiments, this document provides a combination of a MAT2A inhibitor and a topoisomerase inhibitor. In some embodiments, this document provides a combination of a MAT2A inhibitor and SPLAM. In some embodiments, this document provides a combination of a MAT2A inhibitor and a KIF18 inhibitor.

[0113] This disclosure provides a MAT2A inhibitor. In one embodiment, the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I); in X is CH or N; R 3 Halogenated, C 1-6 Halogenated alkyl or C 3-6 cycloalkyl; R 2 For -NR 4 R 5 ; R 4 It is hydrogen or C 1-6 alkyl; R 5 For hydrogen, C 1-6 Alkyl or C 3-6 cycloalkyl; and R 1 It is a phenyl group, wherein the phenyl group is substituted by 0-2 halogen groups.

[0114] In one embodiment, X in formula (I) and its sub-implementations is CH. In another embodiment, X in formula (I) and its sub-implementations is N.

[0115] In another embodiment, R in formula (I) and its sub-implementations 3 Halogenated or C 1-6 Halogenated alkyl. In one embodiment, R in formula (I) and its sub-implementations 3 It is a halogen group. In one embodiment, R in formula (I) and its sub-implementations 3 C 1-6 Halogenated alkyl. In one embodiment, R in formula (I) and its sub-implementations 3 C 3-6 Cycloalkyl. In one embodiment, R in formula (I) and its sub-implementations 3It is chlorine, fluorine, bromine, -CH2Cl, -CF3, -CHF2, -CH2CF3, -CF2CF3, or -CF(CH3)2. In one embodiment, R in formula (I) and its sub-implementations 3 It is chlorine or -CF3. In one embodiment, R in formula (I) and its sub-implementations 3 For chlorine. In one embodiment, R in formula (I) and its sub-implementations 3 It is -CF3.

[0116] In another embodiment, R in formula (I) and its sub-implementations 4 For H. In one implementation, R in formula (I) and its sub-implementations 4 C 1-3 Alkyl group. In one embodiment, R in formula (I) and its sub-implementations 4 It can be methyl, ethyl, propyl or isopropyl.

[0117] In another embodiment, R in formula (I) and its sub-implementations 5 For H. In one implementation, R in formula (I) and its sub-implementations 5 C 1-3 Alkyl group. In one embodiment, R in formula (I) and its sub-implementations 5 It is methyl, ethyl, propyl, or isopropyl. In one embodiment, R in formula (I) and its sub-implementations 5 C 3-6 Cycloalkyl.

[0118] In some implementations of formula (I) and its sub-implementations, R 4 and R 5 Each is H. In some embodiments of equation (I) and its sub-implementations, R 4 and R 5 One of them is H, and the other is C. 1-3 Alkyl group. In some embodiments of formula (I) and its sub-implementations, R 4 and R 5 One of them is H, and the other is methyl, ethyl, propyl, or isopropyl. In some embodiments of formula (I) and its sub-implementations, R 4 and R 5 Independently for C 1-3 alkyl.

[0119] In another embodiment, R in formula (I) and its sub-implementations 2 -NH2, -NHC 1-3 Alkyl or -N(C) 1-3Alkyl)2. In one embodiment, R of formula (I) and its sub-implementations 2 It is NH2, -NHMe, or -N(Me)2. In one embodiment, R in formula (I) and its sub-implementations 2 For NH2. In one embodiment, R in formula (I) and its sub-implementations 2 It is -NHMe.

[0120] In one implementation, R in formula (I) and its sub-implementations 1 It is a phenyl group, which is either unsubstituted or substituted with 1-2 halogen groups. In another embodiment, R in formula (I) and its sub-implementations... 1 The phenyl group is unsubstituted. In one embodiment, R in formula (I) and its sub-implementations... 1 The phenyl group is substituted with one halogen group. In one embodiment, R in formula (I) and its sub-implementations 1 R is a phenyl group substituted with one halogen group selected from fluorine and chlorine. In one embodiment, R in formula (I) and its sub-implementations 1 The phenyl group is substituted with chlorine. In one embodiment, R in formula (I) and its sub-implementations... 1 It is a phenyl group substituted with two halogen groups.

[0121] In another embodiment, the MAT2A inhibitor is selected from the group consisting of compounds in Table 1 or pharmaceutically acceptable salts thereof.

[0122] Table 1.

[0123] In another embodiment, the MAT2A inhibitor is compound A:

[0124] Compound A

[0125] Or its pharmaceutically acceptable salt.

[0126] In another embodiment, the MAT2A inhibitor is compound A1 having the following structural formula:

[0127] Compound A1

[0128] Or its pharmaceutically acceptable salt.

[0129] The preparation and activity of the MAT2A inhibitor described herein are disclosed in PCT / US2019 / 065260 (WO 2020 / 123395), the entire contents of which are hereby incorporated by reference.

[0130] In one embodiment, the combination product comprises a MAT2A inhibitor and a KIF18 inhibitor. In another embodiment, the KIF18 inhibitor is sovexib or a pharmaceutically acceptable salt thereof. The preparation and activity of sovexib are disclosed in PCT / US2019 / 038169 (WO 2020 / 132648), the entire contents of which are hereby incorporated herein by reference in their entirety.

[0131] In another embodiment, the combination product includes a MAT2A inhibitor and SPLAM.

[0132] In another embodiment, SPLAM is indisulan or a pharmaceutically acceptable salt thereof. The preparation and activity of E7820 are disclosed in PCT / JP1994 / 001487 (WO 1995 / 007276) and US 5,767,283, the entire contents of which are hereby incorporated herein by reference.

[0133] In one embodiment, SPLAM is E7820 or a pharmaceutically acceptable salt thereof. The preparation and activity of E7820 are disclosed in PCT / JP2000 / 001071 (WO 2000 / 050395) and US 6,469,043, the entire contents of which are hereby incorporated herein by reference.

[0134] In another embodiment, SPLAM is chloroquinoxaline sulfonamide or a pharmaceutically acceptable salt thereof. The preparation and activity of chloroquinoxaline sulfonamide are disclosed in PCT / US2000 / 000191 (WO 2000 / 040269), the entire contents of which are hereby incorporated herein by reference in their entirety.

[0135] In another embodiment, SPLAM is tacisulan or a pharmaceutically acceptable salt thereof. In another embodiment, SPLAM is sodium tacisulan. The preparation and activity of tacisulan are disclosed in PCT / US2002 / 031568 (WO 2003 / 035629), the entire contents of which are hereby incorporated by reference in their entirety.

[0136] In one implementation, the combined product includes a MAT2A inhibitor and a topoisomerase inhibitor.

[0137] In another embodiment, the topoisomerase inhibitor is a type I topoisomerase inhibitor.

[0138] In another embodiment, the type I topoisomerase inhibitor is 10-hydroxycamptothecin. The preparation and activity of 10-hydroxycamptothecin are disclosed in PCT / US1990 / 005986 (WO 1991 / 005556), the entire contents of which are hereby incorporated herein by reference in their entirety.

[0139] In another embodiment, the type I topoisomerase inhibitor is irinotecan. The preparation and activity of irinotecan are disclosed in PCT / US1992 / 009864 (WO 1993 / 09782), the entire contents of which are hereby incorporated herein by reference in their entirety.

[0140] In one embodiment, the type I topoisomerase inhibitor is topotecan. The preparation and activity of topotecan are disclosed in PCT / US1992 / 001029 (WO 1992 / 014470), the entire contents of which are hereby incorporated by reference in their entirety.

[0141] In one embodiment, the type I topoisomerase inhibitor is hexylresorcinol, ethatecane, delutec, and belotecone, or a pharmaceutically acceptable salt thereof.

[0142] In another embodiment, the topoisomerase inhibitor is a type II topoisomerase inhibitor.

[0143] In another embodiment, the type II topoisomerase inhibitor is daunorubicin. The preparation and activity of daunorubicin are disclosed in US 4,138,480, the entire contents of which are hereby incorporated by reference in their entirety.

[0144] In another embodiment, the type II topoisomerase inhibitor is doxorubicin. The preparation and activity of doxorubicin are disclosed in US 4,138,480, the entire contents of which are hereby incorporated by reference in their entirety.

[0145] In one embodiment, the type II topoisomerase inhibitor is etoposide. The preparation and activity of etoposide are disclosed in PCT / GB1983 / 000257 (WO 1984 / 001506), the entire contents of which are hereby incorporated herein by reference in their entirety.

[0146] Treatment

[0147] In one embodiment, this document provides a method for treating cancer in a subject in need, the method comprising administering to the subject a MAT2A inhibitor and a KIF18 inhibitor, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I); in X is either N or CH; R 3 C 1-6 Halogenated alkyl, halogenated or C 3-6 cycloalkyl; R 2 For -NR 4 R 5 ; R 4 It is hydrogen or C 1-6 alkyl; R 5 For hydrogen, C 1-6 Alkyl or C 3-6 cycloalkyl; and R 1 It is a phenyl group, wherein the phenyl group is substituted by 0-2 halogen groups.

[0148] In another embodiment, this document provides a method for treating cancer in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and a therapeutically effective amount of a KIF18 inhibitor, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0149] In one embodiment, the KIF18 inhibitor is a KIF18A inhibitor. In another embodiment, the KIF18A inhibitor is sovexib or a pharmaceutically acceptable salt thereof. In yet another embodiment, the MAT2A inhibitor is administered simultaneously or sequentially with the KIF18 inhibitor.

[0150] In another embodiment, this document provides a method for treating cancer in a subject in need, the method comprising administering to the subject a MAT2A inhibitor and a splicing inhibitor sulfonamide (SPLAM), wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0151] In another embodiment, this document provides a method for treating cancer in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and a therapeutically effective amount of a splice inhibitor sulfonamide (SPLAM), wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0152] In another embodiment, SPLAM is indisulan or a pharmaceutically acceptable salt thereof. In another embodiment, SPLAM is selected from the group consisting of E7820, chloroquine sulfonamide, and tacisulan or a pharmaceutically acceptable salt thereof. In one embodiment, a MAT2A inhibitor is administered concurrently or sequentially with SPLAM.

[0153] In another embodiment, this document provides a method for treating cancer in a subject in need, the method comprising administering to the subject a MAT2A inhibitor and a topoisomerase inhibitor, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0154] In another embodiment, this document provides a method for treating cancer in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and a therapeutically effective amount of a topoisomerase inhibitor, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0155] In another embodiment, this document provides a method for treating cancer in a subject in need, the method comprising administering to the subject a MAT2A inhibitor and an antibody-drug conjugate (ADC) comprising a topoisomerase inhibitor, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0156] In another embodiment, the topoisomerase inhibitor is a type II topoisomerase inhibitor. In another embodiment, the type I topoisomerase inhibitor is selected from the group consisting of 10-hydroxycamptothecin, irinotecan, and topotecan, or pharmaceutically acceptable salts thereof. In another embodiment, the type I topoisomerase inhibitor is selected from the group consisting of hexylresorcinol, ezetidine, delutec, and belotecidine, or pharmaceutically acceptable salts thereof.

[0157] In another embodiment, the ADC containing a topoisomerase inhibitor is a type I topoisomerase inhibitor. In another embodiment, the ADC containing a topoisomerase inhibitor is trastuzumab (fam-trastuzumabderuxtecan-nxki). In another embodiment, the ADC containing a topoisomerase inhibitor is AZD8205. In another embodiment, the ADC containing a topoisomerase inhibitor is DS-1062 (also known as datopotamabderuxtecan).

[0158] In one embodiment, the topoisomerase inhibitor is a type II topoisomerase inhibitor. In another embodiment, the type II topoisomerase inhibitor is selected from the group consisting of daunorubicin, doxorubicin, and etoposide, or pharmaceutically acceptable salts thereof. In another embodiment, the type II topoisomerase inhibitor is acridine (m-AMSA) or a pharmaceutically acceptable salt thereof. In another embodiment, the MAT2A inhibitor is administered simultaneously or sequentially with the topoisomerase inhibitor.

[0159] In one embodiment of MAT2A of reference formula (I), X is N. In another embodiment, X is CH.

[0160] In another embodiment of MAT2A of reference formula (I), R 4 It is hydrogen and R 5 It is hydrogen or C 1-3 Alkyl group. In another embodiment, R 5 It can be hydrogen or methyl.

[0161] In one implementation of MAT2A of reference formula (I), R 1 It is a phenyl group that has been substituted with chlorine.

[0162] In another embodiment of MAT2A of reference formula (I), R 3 C 1-3 Halogenated alkyl or halogenated. In another embodiment, R 3 It is trifluoromethyl or chlorine. In another embodiment, R 3 It is cyclopropyl.

[0163] In one embodiment, the MAT2A inhibitor is selected from the group consisting of compounds in Table 1 or pharmaceutically acceptable salts thereof. In another embodiment, the MAT2A inhibitor is compound A or a pharmaceutically acceptable salt thereof. In yet another embodiment, the MAT2A inhibitor is compound A1 or a pharmaceutically acceptable salt thereof.

[0164] In one embodiment, this document provides a method for treating cancer in a subject in need, the method comprising administering to the subject a combination comprising a MAT2A inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM) or a KIF18 inhibitor, and at least one pharmaceutically acceptable carrier, thereby treating the subject's cancer. In some embodiments, this document provides a method for treating cancer in a subject in need, the method comprising administering to the subject a combination comprising a MAT2A inhibitor and a topoisomerase inhibitor, and at least one pharmaceutically acceptable carrier, thereby treating the subject's cancer. In some embodiments, this document provides a method for treating cancer in a subject in need, the method comprising administering to the subject a combination comprising a MAT2A inhibitor and a splicing inhibitor sulfonamide (SPLAM), and at least one pharmaceutically acceptable carrier, thereby treating the subject's cancer. In some embodiments, this document provides a method for treating cancer in a subject in need, the method comprising administering to the subject a combination comprising a MAT2A inhibitor and a KIF18 inhibitor, and at least one pharmaceutically acceptable carrier, thereby treating the subject's cancer.

[0165] In another embodiment, the cancer is characterized by reduced or absent expression of the methionine phosphorylase (MTAP) gene, absence of the MTAP gene, reduced function of the MTAP protein, reduced or absent levels of the MTAP protein, accumulation of MTA, or a combination thereof.

[0166] In one embodiment, this document provides a method of treating cancer in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a MAT2A inhibitor and a therapeutically effective amount of a pharmaceutical composition comprising a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor, thereby treating the subject's cancer. In some embodiments, this document provides a method of treating cancer in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a MAT2A inhibitor and a therapeutically effective amount of a pharmaceutical composition comprising a topoisomerase inhibitor, thereby treating the subject's cancer. In some embodiments, this document provides a method of treating cancer in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a MAT2A inhibitor and a therapeutically effective amount of a pharmaceutical composition comprising a splicing inhibitor sulfonamide (SPLAM), thereby treating the subject's cancer. In some embodiments, this document provides a method of treating cancer in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a MAT2A inhibitor and a therapeutically effective amount of a pharmaceutical composition comprising a KIF18 inhibitor, thereby treating the subject's cancer.

[0167] In one embodiment, the use of a combination of a MAT2A inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor is provided for the manufacture of a pharmaceutical agent. In one embodiment, the MAT2A inhibitor is compound A. In one embodiment, the MAT2A inhibitor is compound A1. In one embodiment, the KIF18 inhibitor is sovexib. In another embodiment, the SPLAM is indisulan. In another embodiment, the SPLAM is selected from the group consisting of E7820, chloroquine sulfonamide, and tacisulan, or pharmaceutically acceptable salts thereof. In another embodiment, the topoisomerase inhibitor is selected from the group consisting of 10-hydroxycamptothecin, irinotecan, and topotecan, or pharmaceutically acceptable salts thereof. In another embodiment, the topoisomerase inhibitor is selected from the group consisting of daunorubicin, doxorubicin, and etoposide, or pharmaceutically acceptable salts thereof.

[0168] In one embodiment of the method, the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I); The variables are defined above.

[0169] In another embodiment of the method, the MAT2A inhibitor is selected from the group consisting of compounds in Table 1 or pharmaceutically acceptable salts thereof.

[0170] In another embodiment of the method, the MAT2A inhibitor is compound A or a pharmaceutically acceptable salt thereof.

[0171] In another embodiment of the method, the MAT2A inhibitor is compound A1 or a pharmaceutically acceptable salt thereof.

[0172] In one embodiment of the method, the cancer is selected from the group consisting of: leukemia, glioma, lung cancer, esophageal cancer, MTAP-deficient pancreatic ductal adenocarcinoma (PDAC), melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, anal cancer, gastric cancer, colon cancer, colorectal cancer, soft tissue sarcoma, non-Hodgkin lymphoma, gastric cancer, esophageal gastric cancer, malignant peripheral nerve sheath tumor, mesothelioma, salivary gland tumor, urothelial carcinoma, gastrointestinal cancer, and sarcoma.

[0173] In another embodiment, a product is provided comprising a MAT2A inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor as a combination formulation for simultaneous, single, or sequential medical use. In one embodiment, a product is provided comprising a MAT2A inhibitor and a topoisomerase inhibitor as a combination formulation for simultaneous, single, or sequential medical use. In one embodiment, a product is provided comprising a MAT2A inhibitor and a splicing inhibitor sulfonamide (SPLAM) as a combination formulation for simultaneous, single, or sequential medical use. In another embodiment, a product is provided comprising a MAT2A inhibitor and a KIF18 inhibitor as a combination formulation for simultaneous, single, or sequential medical use. In one embodiment, the MAT2A inhibitor is a compound of formula (I). In one embodiment, the MAT2A inhibitor is a compound in Table 1. In one embodiment, the MAT2A inhibitor is compound A. In one embodiment, the MAT2A inhibitor is compound A1. In one embodiment, the KIF18 inhibitor is sovexicob. In another embodiment, SPLAM is indisulan. In another embodiment, SPLAM is selected from the group consisting of E7820, chloroquine sulfonamide, and tacisulan, or pharmaceutically acceptable salts thereof. In another embodiment, the topoisomerase inhibitor is selected from the group consisting of 10-hydroxycamptothecin, irinotecan, and topotecan, or pharmaceutically acceptable salts thereof. In another embodiment, the topoisomerase inhibitor is selected from the group consisting of daunorubicin, doxorubicin, and etoposide, or pharmaceutically acceptable salts thereof.

[0174] In another embodiment, the cancer is selected from the group consisting of: leukemia, glioma, melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, anal cancer, gastric cancer, colon cancer, colorectal cancer, soft tissue sarcoma, non-Hodgkin's lymphoma, gastric cancer, esophageal cancer, esophageal cancer, malignant peripheral nerve sheath tumor, and mesothelioma.

[0175] In one embodiment, the cancer is mesothelioma. In one embodiment, the cancer is non-small cell lung cancer. In another embodiment, the cancer is non-squamous non-small cell lung cancer. In one embodiment, the cancer is colon or rectal cancer. In one embodiment, the cancer is adenocarcinoma of the colon or rectum. In one embodiment, the cancer is breast cancer. In one embodiment, the cancer is adenocarcinoma of the breast. In one embodiment, the cancer is stomach cancer. In one embodiment, the cancer is gastric adenocarcinoma. In one embodiment, the cancer is pancreatic cancer. In one embodiment, the cancer is pancreatic adenocarcinoma. In one embodiment, the cancer is bladder cancer.

[0176] In one implementation, the cancer is characterized by MTAP ineffectiveness.

[0177] In one implementation, the cancer is characterized as MTAP-deficient.

[0178] In another embodiment, the cancer is a solid tumor. In another embodiment, the cancer is an MTAP-deficient solid tumor. In another embodiment, the cancer is a metastatic MTAP-deficient solid tumor.

[0179] In another embodiment, the cancer is metastatic.

[0180] In another embodiment, the cancer is a solid malignant tumor.

[0181] In another embodiment, the cancer is MTAP-deficient lung cancer or MTAP-deficient pancreatic cancer, including MTAP-deficient NSCLC, MTAP-deficient pancreatic ductal adenocarcinoma (PDAC), or MTAP-deficient esophageal cancer.

[0182] In another implementation, the cancer is a tumor with a deletion of the MTAP gene.

[0183] In any embodiment of this document, the cancer is a solid tumor or a hematologic malignancy. In one embodiment, the tumor is characterized by a lack of MTAP. In another embodiment, the tumor is characterized by normal expression of MTAP.

[0184] In another embodiment, the cancer is NSCLC, mesothelioma, squamous cell carcinoma of the head and neck, salivary gland tumor, urothelial carcinoma, sarcoma, or ovarian cancer. In another embodiment, the cancer is NSCLC, esophageal or gastric cancer, or pancreatic cancer. In yet another embodiment, the cancer is bladder cancer or gastrointestinal cancer.

[0185] In another embodiment, the cancer is characterized by reduced or absent MTAP gene expression, absence of the MTAP gene, reduced function of the MTAP protein, reduced or absent levels of the MTAP protein, MTA accumulation, or a combination thereof.

[0186] In another embodiment, the cancer is characterized by reduced or absent MTAP gene expression.

[0187] In another embodiment, the cancer is characterized by reduced MTAP protein function.

[0188] In another embodiment, the cancer is characterized by reduced or absent MTAP protein levels.

[0189] In another embodiment, the cancer is characterized by MTA accumulation.

[0190] In another embodiment, the cancer is a tumor with PRKC fusion.

[0191] In one embodiment, this article provides a method for inhibiting tumor growth or slowing the tumor growth rate in a subject with MTAP-deficient cancer, the method comprising administering a therapeutically effective amount of a MAT2A inhibitor and a therapeutically effective amount of a topoisomerase inhibitor.

[0192] In another embodiment, this document provides a method for inhibiting tumor growth or slowing tumor growth rate in a subject with MTAP-deficient cancer, the method comprising administering a therapeutically effective amount of a MAT2A inhibitor and a therapeutically effective amount of SPLAM.

[0193] In another embodiment, this document provides a method for inhibiting tumor growth or slowing the tumor growth rate in a subject with MTAP-deficient cancer, the method comprising administering a therapeutically effective amount of a MAT2A inhibitor and a therapeutically effective amount of a KIF18 inhibitor.

[0194] Tumor growth is typically measured by changes in tumor volume acquired from a first time point to a second time point. In one embodiment, tumor growth is measured by changes in tumor volume from the first time point to the second time point. In some embodiments, the tumor volume at the second time point does not increase compared to the first time point. In some embodiments, the tumor volume decreases from the first time point to the second time point.

[0195] In one embodiment, this document provides a method for treating cancer in a subject of need, the method comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and a therapeutically effective amount of a topoisomerase inhibitor, wherein the subject has not previously received MAT2A inhibitor treatment. In some embodiments, the treatment reduces tumor growth rate compared to treatment with a MAT2A inhibitor alone for a similar time period.

[0196] In another embodiment, this document provides a method for treating cancer in a subject of need, the method comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and a therapeutically effective amount of SPLM, wherein the subject has not previously received MAT2A inhibitor treatment. In some embodiments, the treatment reduces the tumor growth rate compared to treatment with a MAT2A inhibitor alone for a similar time period.

[0197] In another embodiment, this document provides a method for treating cancer in a subject of need, the method comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and a therapeutically effective amount of KIF8, wherein the subject has not previously received MAT2A inhibitor treatment. In some embodiments, the treatment reduces the tumor growth rate compared to treatment with a MAT2A inhibitor alone for a similar time period.

[0198] In one embodiment, the MAT2A inhibitor and the topoisomerase inhibitor, the splicing inhibitor sulfonamide (SPLAM), or the KIF18 inhibitor are in separate dosage forms. In another embodiment, the MAT2A inhibitor and the topoisomerase inhibitor are in separate dosage forms. In yet another embodiment, the MAT2A inhibitor and the KIF18 inhibitor are in separate dosage forms.

[0199] In another embodiment, the MAT2A inhibitor and the topoisomerase inhibitor, the splicing inhibitor sulfonamide (SPLAM), or the KIF18 inhibitor are in the same dosage form. In one embodiment, the MAT2A inhibitor and the topoisomerase inhibitor are in the same dosage form. In one embodiment, the MAT2A inhibitor and the splicing inhibitor sulfonamide (SPLAM) are in the same dosage form. In another embodiment, the MAT2A inhibitor and the KIF18 inhibitor are in the same dosage form.

[0200] In another embodiment, the treatment comprises substantially simultaneous administration of a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof. In another embodiment, the treatment comprises administration at different times of a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof.

[0201] In another embodiment, an individual is given a MAT2A inhibitor or a pharmaceutically acceptable salt thereof, followed by the administration of a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof. In one embodiment, an individual is given a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof, followed by the administration of a MAT2A inhibitor or a pharmaceutically acceptable salt thereof.

[0202] In some embodiments, the MAT2A inhibitor and the topoisomerase inhibitor are administered simultaneously. In some embodiments, the MAT2A inhibitor and the topoisomerase inhibitor are administered sequentially. In some embodiments, the MAT2A inhibitor is administered before the topoisomerase inhibitor. In some embodiments, the MAT2A inhibitor is administered after the topoisomerase inhibitor.

[0203] In some embodiments, the MAT2A inhibitor and SPLAM are administered simultaneously. In some embodiments, the MAT2A inhibitor and SPLAM are administered sequentially. In some embodiments, the MAT2A inhibitor is administered before SPLAM. In some embodiments, the MAT2A inhibitor is administered after SPLAM.

[0204] In some embodiments, the MAT2A inhibitor and the KIF18 inhibitor are administered simultaneously. In some embodiments, the MAT2A inhibitor and the KIF18 inhibitor are administered sequentially. In some embodiments, the MAT2A inhibitor is administered before the KIF18 inhibitor. In some embodiments, the MAT2A inhibitor is administered after the KIF18 inhibitor.

[0205] In another embodiment, the method includes administering a MAT2A inhibitor to a subject in need.

[0206] In another embodiment, the method includes administering a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor to a subject in need.

[0207] In one embodiment, a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof are administered orally. In one embodiment, a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a topoisomerase inhibitor or a pharmaceutically acceptable salt thereof are each administered orally. In one embodiment, a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a SPLAM or a pharmaceutically acceptable salt thereof are each administered orally. In one embodiment, a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a KIF18 inhibitor or a pharmaceutically acceptable salt thereof are each administered orally.

[0208] In another implementation, the cancer to be treated is selected from the group consisting of: leukemia, glioma, melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, anal cancer, gastric cancer, colon cancer, colorectal cancer, soft tissue sarcoma, non-Hodgkin's lymphoma, gastric cancer, esophageal cancer, esophageal cancer, malignant peripheral nerve sheath tumor, and mesothelioma.

[0209] In one aspect, this article provides a MAT2A inhibitor or a pharmaceutically acceptable salt thereof, and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM) or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof, for treatment.

[0210] In one implementation, a MAT2A inhibitor or a pharmaceutically acceptable salt thereof, and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof are used to treat cancer in a subject in need.

[0211] Exemplary durations of treatment associated with the treatment methods disclosed herein include: approximately one week; approximately two weeks; approximately three weeks; approximately four weeks; approximately five weeks; approximately six weeks; approximately seven weeks; approximately eight weeks; approximately nine weeks; approximately ten weeks; approximately eleven weeks; approximately twelve weeks; approximately thirteen weeks; approximately fourteen weeks; approximately fifteen weeks; approximately sixteen weeks; approximately seventeen weeks; approximately eighteen weeks; approximately nineteen weeks; approximately twenty-one weeks; approximately twenty-two weeks; approximately twenty-three weeks; approximately twenty-four weeks; approximately seven months; approximately eight months; approximately nine months; approximately ten months; approximately eleven months; approximately twelve months; approximately thirteen months; approximately fourteen months; approximately fifteen months; approximately sixteen months; approximately seventeen months; approximately eighteen months; approximately nineteen months; approximately twenty months; approximately twenty-one months; approximately twenty-two months; approximately twenty-three months; approximately twenty-four months; approximately thirty months; approximately three years; approximately four years; and approximately five years.

[0212] In one embodiment of the method, the method involves administering a therapeutically effective amount of a combination or composition comprising the compounds provided herein or pharmaceutically acceptable salts thereof to a subject requiring treatment (including subjects identified as needing treatment) (including, but not limited to, humans or animals).

[0213] In another embodiment of the method, the treatment comprises co-administering a certain amount of a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a certain amount of a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor or a pharmaceutically acceptable salt thereof. In one embodiment, a certain amount of a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a certain amount of a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor or a pharmaceutically acceptable salt thereof are in a single formulation or unit dosage form. In other embodiments, a certain amount of a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a certain amount of a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor or a pharmaceutically acceptable salt thereof are in a separate formulation or unit dosage form.

[0214] In the foregoing methods, treatment may include substantially simultaneous administration of a certain amount of a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a certain amount of a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor or a pharmaceutically acceptable salt thereof, or administration of a certain amount of a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a certain amount of a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor or a pharmaceutically acceptable salt thereof at different times. In some embodiments of the foregoing methods, the certain amount of a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and / or the certain amount of a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor or a pharmaceutically acceptable salt thereof is administered at a dose that is ineffective when administered alone as a combination of the MAT2A inhibitor or a pharmaceutically acceptable salt thereof and one or both of the topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor or a pharmaceutically acceptable salt thereof, but effective when the amounts are used in combination.

[0215] Pharmaceutical Composition

[0216] In one aspect, this article provides a pharmaceutical composition comprising a MAT2A inhibitor or a pharmaceutically acceptable salt thereof, a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM) or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier.

[0217] In one embodiment, a pharmaceutical composition is provided comprising a therapeutically effective amount of a MAT2A inhibitor or a pharmaceutically acceptable salt thereof, and a second pharmaceutical composition comprising a therapeutically effective amount of a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof.

[0218] In another embodiment, this document provides a combination product comprising a first pharmaceutical composition comprising a therapeutically effective amount of a MAT2A inhibitor or a pharmaceutically acceptable salt thereof, and a second pharmaceutical composition comprising a therapeutically effective amount of a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof.

[0219] In some embodiments, the second pharmaceutical composition comprises a topoisomerase inhibitor. In some embodiments, the second pharmaceutical composition comprises SPLAM. In some embodiments, the second pharmaceutical composition comprises a KIF18 inhibitor.

[0220] In one embodiment, the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I); The variables are defined above.

[0221] In another embodiment, the MAT2A inhibitor is selected from the group consisting of compounds in Table 1 or pharmaceutically acceptable salts thereof.

[0222] In another embodiment, the MAT2A inhibitor is compound A:

[0223] Compound A

[0224] Or its pharmaceutically acceptable salt.

[0225] In another embodiment, the MAT2A inhibitor is compound A1:

[0226] Compound A1

[0227] Or its pharmaceutically acceptable salt.

[0228] In one embodiment, the KIF18 inhibitor is sovexib. In another embodiment, SPLAM is indisulan. In another embodiment, SPLAM is selected from the group consisting of E7820, chloroquine sulfonamide, and tacisulan, or pharmaceutically acceptable salts thereof. In another embodiment, the topoisomerase inhibitor is selected from the group consisting of 10-hydroxycamptothecin, irinotecan, and topotecan, or pharmaceutically acceptable salts thereof. In another embodiment, the topoisomerase inhibitor is selected from the group consisting of daunorubicin, doxorubicin, and etoposide, or pharmaceutically acceptable salts thereof.

[0229] In another embodiment, this document provides a combination product comprising a first pharmaceutical composition comprising a therapeutically effective amount of compound A or a pharmaceutically acceptable salt thereof; and a second pharmaceutical composition comprising a therapeutically effective amount of a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof. In some embodiments, the second pharmaceutical composition comprises a topoisomerase inhibitor. In some embodiments, the second pharmaceutical composition comprises SPLAM. In some embodiments, the second pharmaceutical composition comprises a KIF18 inhibitor.

[0230] In one embodiment, the pharmaceutical composition is used to treat a patient's cancer. In one embodiment, the pharmaceutical composition is used to treat a patient's solid tumor. In one embodiment, the pharmaceutical composition is used to treat a patient's solid malignant tumor. In another embodiment, the cancer is MTAP-deficient lung cancer or MTAP-deficient pancreatic cancer, including MTAP-deficient NSCLC, MTAP-deficient PDAC, or MTAP-deficient esophageal cancer.

[0231] In any embodiment of this document, the cancer is a solid tumor or a hematologic malignancy. In another embodiment, the cancer is NSCLC, mesothelioma, squamous cell carcinoma of the head and neck, salivary gland tumor, urothelial carcinoma, sarcoma, or ovarian cancer. In another embodiment, the cancer is NSCLC, esophageal or gastric cancer, or pancreatic cancer. In another embodiment, the cancer is bladder cancer or gastrointestinal cancer. In another embodiment, the cancer is bladder cancer (urothelial carcinoma) or gastrointestinal cancer. In one embodiment, the cancer is bladder cancer. In another embodiment, the cancer is urothelial carcinoma. In another embodiment, the cancer is gastrointestinal cancer.

[0232] In one embodiment, the pharmaceutical composition is used to treat a patient with mesothelioma. In one embodiment, the pharmaceutical composition is used to treat a patient with non-small cell lung cancer. In one embodiment, the pharmaceutical composition is used to treat a patient with non-squamous non-small cell lung cancer. In one embodiment, the pharmaceutical composition is used to treat a patient with colon cancer. In one embodiment, the pharmaceutical composition is used to treat a patient with rectal cancer. In one embodiment, the pharmaceutical composition is used to treat a patient with colonic adenocarcinoma or rectal adenocarcinoma. In one embodiment, the pharmaceutical composition is used to treat a patient with breast cancer. In one embodiment, the pharmaceutical composition is used to treat a patient with breast adenocarcinoma. In one embodiment, the pharmaceutical composition is used to treat a patient with gastric cancer. In one embodiment, the pharmaceutical composition is used to treat a patient with gastric adenocarcinoma. In one embodiment, the pharmaceutical composition is used to treat a patient with pancreatic cancer. In one embodiment, the pharmaceutical composition is used to treat a patient with pancreatic adenocarcinoma. In one embodiment, the pharmaceutical composition is used to treat a patient with bladder cancer.

[0233] Application / Dosage / Formulation

[0234] Administering the combination includes administering the combination as a single formulation or unit dosage form, administering individual agents of the combination simultaneously but separately, or administering individual agents of the combination sequentially via any suitable route. The dosage of a single agent in the combination may require more frequent administration of one or more of the other agents in the combination. Therefore, to allow for proper dosing, packaged pharmaceutical products may include one or more dosage forms containing a combination of agents, and one or more dosage forms containing one agent of a combination but without the other agents in the combination.

[0235] The actual dose level of the active ingredient in a pharmaceutical composition can be varied to obtain an amount of the active ingredient that effectively achieves the desired therapeutic response for a particular patient, composition, and administration mode without being toxic to the patient.

[0236] In particular, the selected dose level will depend on a variety of factors, including the activity of the specific compound used, the timing of administration, the rate of excretion of the compound, the duration of treatment, other drugs, compounds or materials used in combination with the compound, the age, sex, weight, condition, general health status and medical history of the patient being treated, and similar factors well known in the medical field.

[0237] A physician or veterinarian with the skills in the art can easily determine and prescribe an effective amount of the desired pharmaceutical composition. For example, a physician or veterinarian may begin administering the pharmaceutical composition at a level lower than required to achieve the desired therapeutic effect, and gradually increase the dose until the desired effect is achieved.

[0238] In certain embodiments, it is particularly advantageous to formulate the compound in unitary dosage form for ease of administration and uniform dosage. As used herein, unitary dosage form refers to a physically discrete unit suitable as a single dose for a patient to be treated; each unit contains a predetermined amount of the disclosed compound bound to the desired pharmaceutical medium in a manner calculated to produce the desired therapeutic effect. The unitary dosage form is determined by and directly depends on (a) the unique properties of the disclosed compound and the specific therapeutic effect to be achieved, and (b) the inherent limitations in the field of coagulating / formulating such a disclosed compound for the treatment of a patient's pain, depressive disorder, or drug addiction.

[0239] In one embodiment, the compound provided herein is formulated using one or more pharmaceutically acceptable excipients or carriers. In one embodiment, the pharmaceutical composition provided herein comprises a therapeutically effective amount of the disclosed compound and a pharmaceutically acceptable carrier.

[0240] The optimal proportions, individual doses, and combined doses and concentrations of pharmaceutical compounds that produce efficacy without toxicity are determined based on the kinetics of the utilization of the active ingredient against the target site and using methods known to those skilled in the art.

[0241] Routes of administration for any of the compositions discussed herein include oral, nasal, rectal, vaginal, parenteral, buccal, sublingual, or topical. The compounds can be formulated for administration via any suitable route, such as oral or parenteral, such as transdermal, transmucosal (e.g., sublingual, translingual, (trans)buccal, (trans)urethral, ​​vaginal (e.g., vaginal and perivallary), (internal)nasal, and (trans)rectal), intravesical, intrapulmonary, intraduodenal, intragastric, intrathecal, intrathecal, subcutaneous, intramuscular, intradermal, intraarterial, intravenous, intrabronchial, inhalation, and topical administration. In one embodiment, the preferred route of administration is oral.

[0242] Suitable compositions and dosage forms include, for example, tablets, capsules, granules, pills, gel capsules, lozenges, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pills, pastes, lozenges, creams, ointments, lotions, tablets, suppositories, liquid sprays for nasal or oral administration, dry powder or nebulized formulations for inhalation, and compositions and formulations for intravesical administration. It should be understood that formulations and compositions are not limited to the specific formulations and compositions described herein.

[0243] For oral applications, tablets, sugar-coated pills, liquids, drops, suppositories or capsules, capsules, and gel capsules are particularly suitable. Compositions intended for oral use can be prepared according to any method known in the art, and such compositions may contain one or more agents selected from the group consisting of inert, non-toxic pharmaceutically effective excipients suitable for the manufacture of tablets. Such excipients include, for example, inert diluents such as lactose; granulating and disintegrants such as corn starch; binders such as starch; and lubricants such as magnesium stearate. Tablets may be uncoated or may be coated using known techniques for aesthetic purposes or to delay the release of the active ingredient. Formulations for oral use may also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert diluent.

[0244] For parenteral administration, the disclosed compounds can be formulated for injection or infusion, such as intravenous, intramuscular, or subcutaneous injection or infusion, or for administration by bolus dose or continuous infusion. Suspensions, solutions, or emulsions in oily or aqueous media may be used, optionally containing other formulations such as suspending agents, stabilizers, or dispersants.

[0245] Reagent test kit

[0246] In one aspect, this disclosure provides a kit for treating cancer, comprising a MAT2A inhibitor or a pharmaceutically acceptable salt thereof, and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM) or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof.

[0247] In some embodiments, the kit includes a pharmaceutical product comprising a pharmaceutical composition comprising a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or diluent; and a pharmaceutical composition comprising a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM) or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or diluent.

[0248] In some embodiments, the kit comprises a pharmaceutical composition containing a MAT2A inhibitor or a pharmaceutically acceptable salt thereof; a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM) or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or diluent.

[0249] In another embodiment, a pharmaceutical kit is provided. The kit includes a sealed container approved for storing a pharmaceutical composition containing one of the aforementioned pharmaceutical compositions. In some embodiments, the sealed container minimizes contact between air and the ingredients, such as a vacuum bottle. In other embodiments, the sealed container is a sealed tube. The kit should include instructions for use of the composition and information about the composition.

[0250] In certain embodiments, the combined compounds can be administered on the same schedule, either by administering a single formulation or unit containing all the compounds in the combination, or by administering a separate formulation or unit containing one of the compounds in the combination. However, some compounds used in the combination may be administered more frequently than once daily, or at a different frequency than the other compounds in the combination. Therefore, in one embodiment, the kit contains in a container a formulation or unit containing all the compounds in the combination, and an additional formulation or unit containing one of the compounds in the inclusion agent combination, without any additional active compounds, and includes instructions for administering the formulation on a fixed schedule.

[0251] The kits described herein include prescription information, such as prescriptions to patients or healthcare providers, or labels on packaged pharmaceutical preparations. Prescription information may include, for example, information regarding the efficacy, dosage and administration, contraindications, and adverse reactions of the pharmaceutical preparation.

[0252] In all the foregoing cases, the compound combinations of the present invention may be applied alone, as a mixture, or together with other active agents.

[0253] The kits provided herein are designed to maintain the conditions necessary for properly maintaining the components contained therein (e.g., refrigeration or freezing). The kits may include labels or instructions for use, including identification information for the components and instructions for use (e.g., dosing parameters, clinical pharmacology of the active ingredient, including mechanism of action, pharmacokinetics and pharmacodynamics, adverse reactions, contraindications, etc.).

[0254] The components of the kit can be packaged in individual containers, and all different containers can be contained in a single package. Labels or instructions may include manufacturer information such as batch number and expiration date. Labels or instructions may be integrated into the physical structure containing the components, contained separately within the physical structure, or affixed to components of the kit (e.g., ampoules, syringes, or vials).

[0255] Non-limiting exemplary implementation ： Part A - Non-limiting exemplary embodiments 1 to 112 In the further embodiments 1 to 112 described below, this disclosure includes: Implementation Scheme 1. In Implementation Scheme 1, a method for treating cancer in a subject of need is provided, the method comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and administering to the subject a therapeutically effective amount of a KIF18 inhibitor.

[0256] Implementation Scheme 2. In Implementation Scheme 2, a method for treating cancer in a subject in need is provided, the method comprising administering a therapeutically effective amount of a MAT2A inhibitor to the subject, wherein the subject has already received treatment with a KIF18 inhibitor.

[0257] Implementation Scheme 3. In Implementation Scheme 3, a method for treating cancer in a subject of need is provided, the method comprising administering a therapeutically effective amount of a MAT2A inhibitor to the subject, wherein the subject is simultaneously receiving a KIF18 inhibitor.

[0258] Implementation Scheme 4. In Implementation Scheme 4, a method according to any one of Implementation Schemes 1 to 3 is provided, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I); in X is either N or CH; R 3 C 1-6 Halogenated alkyl, halogenated or C 3-6 cycloalkyl; R 2 For -NR 4 R 5 ; R 4 It is hydrogen or C 1-6 alkyl; R 5 For hydrogen, C 1-6 Alkyl or C 3-6 cycloalkyl; and R 1 It is a phenyl group, wherein the phenyl group is substituted by 0-2 halogen groups.

[0259] Implementation Scheme 5. In Implementation Scheme 5, a method according to any one of Implementation Schemes 1 to 4 is provided, wherein the KIF18 inhibitor is a KIF18A inhibitor.

[0260] Implementation Scheme 6. In Implementation Scheme 6, a method according to any one of Implementation Schemes 1 to 5 is provided, wherein the KIF18A inhibitor is sovexib (compound B) or a pharmaceutically acceptable salt thereof.

[0261] Implementation Scheme 7. In Implementation Scheme 7, a method according to any one of Implementation Schemes 1 to 6 is provided, wherein the MAT2A inhibitor and the KIF18 inhibitor are administered simultaneously or sequentially.

[0262] Implementation Scheme 8. In Implementation Scheme 8, a method for treating cancer in a subject of need is provided, the method comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and administering to the subject a therapeutically effective amount of a splicing inhibitor sulfonamide (SPLAM).

[0263] Implementation Scheme 9. In Implementation Scheme 9, a method for treating cancer in a subject of need is provided, the method comprising administering a therapeutically effective amount of a MAT2A inhibitor to the subject, wherein the subject has already received treatment with the splice inhibitor sulfonamide (SPLAM).

[0264] Implementation Scheme 10. In Implementation Scheme 10, a method for treating cancer in a subject of need is provided, the method comprising administering a therapeutically effective amount of a MAT2A inhibitor to the subject, wherein the subject simultaneously receives a splicing inhibitor sulfonamide (SPLAM).

[0265] Implementation Scheme 11. In Implementation Scheme 11, a method according to any one of Implementation Schemes 8 to 10 is provided, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I); in X is either N or CH; R 3 C 1-6 Halogenated alkyl, halogenated or C 3-6 cycloalkyl; R 2 For -NR 4 R 5 ; R 4 It is hydrogen or C 1-6 alkyl; R 5 For hydrogen, C 1-6 Alkyl or C 3-6 cycloalkyl; and R 1 It is a phenyl group, wherein the phenyl group is substituted by 0-2 halogen groups.

[0266] Implementation Scheme 12. In Implementation Scheme 12, a method according to any one of Implementation Schemes 8 to 11 is provided, wherein the SPLAM is indisulan or a pharmaceutically acceptable salt thereof.

[0267] Implementation Scheme 13. In Implementation Scheme 13, a method according to any one of Implementation Schemes 8 to 11 is provided, wherein the SPLAM is selected from the group consisting of: E7820, chloroquine sulfonamide, and tacisulan or a pharmaceutically acceptable salt thereof.

[0268] Implementation Scheme 14. In Implementation Scheme 14, a method according to any one of Implementation Schemes 8 to 13 is provided, wherein the MAT2A inhibitor is administered simultaneously or sequentially with SPLAM.

[0269] Implementation Scheme 15. In Implementation Scheme 15, a method for treating cancer in a subject of need is provided, the method comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and administering to the subject a therapeutically effective amount of a topoisomerase inhibitor.

[0270] Implementation Scheme 16. In Implementation Scheme 16, a method for treating cancer in a subject in need is provided, the method comprising administering a therapeutically effective amount of a MAT2A inhibitor to the subject, wherein the subject has already received treatment with a topoisomerase inhibitor.

[0271] Implementation Scheme 17. In Implementation Scheme 17, a method for treating cancer in a subject of need is provided, the method comprising administering a therapeutically effective amount of a MAT2A inhibitor to the subject, wherein the subject simultaneously receives a topoisomerase inhibitor.

[0272] Implementation Scheme 18. In Implementation Scheme 18, a method according to any one of Implementation Schemes 15 to 17 is provided, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I); in X is either N or CH; R 3 C 1-6 Halogenated alkyl, halogenated or C 3-6 cycloalkyl; R 2 For -NR 4 R 5 ; R 4 It is hydrogen or C 1-6 alkyl; R 5 For hydrogen, C 1-6 Alkyl or C 3-6 cycloalkyl; and R 1 It is a phenyl group, wherein the phenyl group is substituted by 0-2 halogen groups.

[0273] Implementation Scheme 19. In Implementation Scheme 19, a method according to any one of Implementation Schemes 15 to 18 is provided, wherein the topoisomerase inhibitor is a type I topoisomerase inhibitor.

[0274] Implementation Scheme 20. In Implementation Scheme 20, a method according to Implementation Scheme 19 is provided, wherein the type I topoisomerase inhibitor is selected from the group consisting of 10-hydroxycamptothecin, irinotecan and topotecan, or pharmaceutically acceptable salts thereof.

[0275] Implementation Scheme 20A. In Implementation Scheme 20A, the method according to Implementation Scheme 19 is provided, wherein the type I topoisomerase inhibitor is selected from the group consisting of: hexylresorcinol, ethatecan, delutecan and belotecone, or pharmaceutically acceptable salts thereof.

[0276] Implementation Scheme 21. In Implementation Scheme 21, a method according to any one of Implementation Schemes 15 to 18 is provided, wherein the topoisomerase inhibitor is a type II topoisomerase inhibitor.

[0277] Implementation Scheme 22. In Implementation Scheme 22, the method according to Implementation Scheme 21 is provided, wherein the type II topoisomerase inhibitor is selected from the group consisting of: daunorubicin, doxorubicin and etoposide, or pharmaceutically acceptable salts thereof.

[0278] Implementation Scheme 23. In Implementation Scheme 23, a method according to any one of Implementation Schemes 15 to 22 is provided, wherein the MAT2A inhibitor and the topoisomerase inhibitor are administered simultaneously or sequentially.

[0279] Implementation Scheme 24. In Implementation Scheme 24, a method according to any one of Implementation Schemes 4 to 7, 11 to 14 and 18 to 23 is provided, wherein X is N.

[0280] Implementation Scheme 25. In Implementation Scheme 25, a method according to any one of Implementation Schemes 4 to 7, 11 to 14 and 18 to 23 is provided, wherein X is CH.

[0281] Implementation Scheme 26. In Implementation Scheme 26, a method according to any one of Implementation Schemes 4 to 7, 11 to 14 and 18 to 25 is provided, wherein R 4 It is hydrogen and R 5 It is hydrogen or C 1-3 alkyl.

[0282] Implementation Scheme 27. In Implementation Scheme 27, a method according to any one of Implementation Schemes 4 to 7, 11 to 14 and 18 to 26 is provided, wherein R 5 It can be hydrogen or methyl.

[0283] Implementation Scheme 28. In Implementation Scheme 28, a method according to any one of Implementation Schemes 4 to 7, 11 to 14 and 18 to 27 is provided, wherein R 1 It is a phenyl group that has been substituted with chlorine.

[0284] Implementation Scheme 29. In Implementation Scheme 29, a method according to any one of Implementation Schemes 4 to 7, 11 to 14 and 18 to 28 is provided, wherein R 3 C 1-3 Halogenated alkyl or halogenated.

[0285] Implementation Scheme 30. In Implementation Scheme 30, a method according to any one of Implementation Schemes 4 to 7, 11 to 14 and 18 to 29 is provided, wherein R 3 It is trifluoromethyl or chlorine.

[0286] Implementation Scheme 31. In Implementation Scheme 31, a method according to any one of Implementation Schemes 4 to 7, 11 to 14 and 18 to 28 is provided, wherein R 3 It is cyclopropyl.

[0287] Implementation Scheme 32. In Implementation Scheme 32, a method according to any one of Implementation Schemes 1 to 31 is provided, wherein the MAT2A inhibitor is selected from the group consisting of compounds in Table 1 or pharmaceutically acceptable salts thereof.

[0288] Implementation Scheme 33. In Implementation Scheme 33, a method according to any one of Implementation Schemes 1 to 32 is provided, wherein the MAT2A inhibitor is compound A or a pharmaceutically acceptable salt thereof.

[0289] Implementation Scheme 34. In Implementation Scheme 34, a method according to any one of Implementation Schemes 1 to 32 is provided, wherein the MAT2A inhibitor is compound A1 or a pharmaceutically acceptable salt thereof.

[0290] Implementation Scheme 35. In Implementation Scheme 35, a method according to any one of Implementation Schemes 1 to 34 is provided, wherein the cancer is selected from the group consisting of: leukemia, glioma, lung cancer, esophageal cancer, MTAP-deficient pancreatic ductal adenocarcinoma (PDAC), melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, anal cancer, gastric cancer, colon cancer, colorectal cancer, soft tissue sarcoma, non-Hodgkin lymphoma, gastric cancer, esophageal cancer, esophageal cancer, malignant peripheral nerve sheath tumor, mesothelioma, salivary gland tumor, urothelial carcinoma, gastrointestinal cancer, and sarcoma.

[0291] Implementation Scheme 36. In Implementation Scheme 36, a method according to any one of Implementation Schemes 1 to 35 is provided, wherein the cancer is a solid tumor or a hematologic cancer.

[0292] Implementation Scheme 37. In Implementation Scheme 37, a method according to any one of Implementation Schemes 1 to 36 is provided, wherein the cancer is a solid tumor.

[0293] Implementation Scheme 38. In Implementation Scheme 38, a method according to any one of Implementation Schemes 1 to 37 is provided, wherein the cancer is a solid malignant tumor.

[0294] Implementation Scheme 38A. In Implementation Scheme 38A, a method according to any one of Implementation Schemes 1 to 37 is provided, wherein the cancer is characterized by reduced or absent expression of the MTAP gene, absence of the MTAP gene, reduced function of the MTAP protein, reduced or absent level of the MTAP protein, accumulation of MTA, or a combination thereof.

[0295] Implementation Scheme 39. In Implementation Scheme 39, a method is provided according to any one of Implementation Schemes 1 to 38 and 38A, wherein the cancer is characterized by the deletion of the MTAP gene.

[0296] Implementation Scheme 40. In Implementation Scheme 40, the use of a MAT2A inhibitor in the manufacture of a medicament for the treatment of cancer is provided, wherein the MAT2A inhibitor is administered simultaneously or sequentially with a KIF18 inhibitor.

[0297] Implementation Scheme 41. In Implementation Scheme 41, the KIF18 inhibitor described in Implementation Scheme 40 is a KIF18A inhibitor.

[0298] Implementation Scheme 42. In Implementation Scheme 42, the KIF18A inhibitor described in Implementation Scheme 41 is, wherein the KIF18A inhibitor is sovexib (compound B) or a pharmaceutically acceptable salt thereof.

[0299] Implementation Scheme 43. In Implementation Scheme 43, the use of a MAT2A inhibitor in the manufacture of a medicament for the treatment of cancer is provided, wherein the MAT2A inhibitor is administered simultaneously or sequentially with SPLAM.

[0300] Implementation Scheme 44. In Implementation Scheme 44, the SPLAM described in Implementation Scheme 43, wherein the SPLAM is indisulan or a pharmaceutically acceptable salt thereof.

[0301] Implementation Scheme 45. In Implementation Scheme 45, the SPLAM described in Implementation Scheme 43, wherein the SPLAM is selected from the group consisting of: E7820, chloroquine sulfonamide, and tacisulan or a pharmaceutically acceptable salt thereof.

[0302] Implementation Scheme 46. In Implementation Scheme 46, the use of a MAT2A inhibitor in the manufacture of a medicament for the treatment of cancer is provided, wherein the MAT2A inhibitor is administered simultaneously or sequentially with a topoisomerase inhibitor.

[0303] Implementation Scheme 47. In Implementation Scheme 47, the topoisomerase inhibitor described in Implementation Scheme 46, wherein the topoisomerase inhibitor is a type I topoisomerase inhibitor.

[0304] Implementation Scheme 48. In Implementation Scheme 48, the type I topoisomerase inhibitor described in Implementation Scheme 47, wherein the type I topoisomerase inhibitor is selected from the group consisting of 10-hydroxycamptothecin, irinotecan and topotecan, or pharmaceutically acceptable salts thereof.

[0305] Implementation Scheme 48A. In Implementation Scheme 48A, the use according to Implementation Scheme 47 is provided, wherein the type I topoisomerase inhibitor is selected from the group consisting of: hexylresorcinol, ethatecan, delutecan and belotecone, or pharmaceutically acceptable salts thereof.

[0306] Implementation Scheme 49. In Implementation Scheme 49, the topoisomerase inhibitor described in Implementation Scheme 48, wherein the topoisomerase inhibitor is a type II topoisomerase inhibitor.

[0307] Implementation Scheme 50. In Implementation Scheme 50, the type II topoisomerase inhibitor described in Implementation Scheme 49, wherein the type II topoisomerase inhibitor is selected from the group consisting of: daunorubicin, doxorubicin and etoposide, or pharmaceutically acceptable salts thereof.

[0308] Implementation Scheme 51. In Implementation Scheme 51, the use described in any one of Implementation Schemes 40 to 50 is provided, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I); in X is either N or CH; R 3 C 1-6 Halogenated alkyl, halogenated or C 3-6 cycloalkyl; R 2 For -NR 4 R 5 ; R4 It is hydrogen or C 1-6 alkyl; R 5 For hydrogen, C 1-6 Alkyl or C 3-6 cycloalkyl; and R 1 It is a phenyl group, wherein the phenyl group is substituted by 0-2 halogen groups.

[0309] Implementation Scheme 52. In Implementation Scheme 52, the use of the MAT2A inhibitor described in Implementation Scheme 51 is provided, wherein the MAT2A inhibitor is a compound of formula (I) as defined in any one of Implementation Schemes 24 to 31 or a pharmaceutically acceptable salt thereof.

[0310] Implementation Scheme 53. In Implementation Scheme 53, the use of the MAT2A inhibitor described in any one of Implementation Schemes 40 to 52 is provided, wherein the MAT2A inhibitor is selected from the group consisting of compounds in Table 1 or pharmaceutically acceptable salts thereof.

[0311] Implementation Scheme 54. In Implementation Scheme 54, the use of the MAT2A inhibitor described in any one of Implementation Schemes 40 to 53 is provided, wherein the MAT2A inhibitor is compound A or a pharmaceutically acceptable salt thereof.

[0312] Implementation Scheme 55. In Implementation Scheme 55, the use of the MAT2A inhibitor described in any one of Implementation Schemes 40 to 53 is provided, wherein the MAT2A inhibitor is a group consisting of compound A1 or a pharmaceutically acceptable salt thereof.

[0313] Implementation Scheme 56. In Implementation Scheme 56, a MAT2A inhibitor for treating cancer is provided, wherein the MAT2A inhibitor is administered simultaneously or sequentially with a KIF18 inhibitor.

[0314] Implementation Scheme 57. In Implementation Scheme 57, the KIF18 inhibitor described in Implementation Scheme 57 is a KIF18A inhibitor.

[0315] Implementation Scheme 58. In Implementation Scheme 58, the KIF18A inhibitor described in Implementation Scheme 57 is, wherein the KIF18A inhibitor is sovexib (compound B) or a pharmaceutically acceptable salt thereof.

[0316] Implementation Scheme 59. In Implementation Scheme 59, a MAT2A inhibitor for treating cancer is provided, wherein the MAT2A inhibitor is administered simultaneously or sequentially with SPLAM.

[0317] Implementation Scheme 60. In Implementation Scheme 60, the SPLAM described in Implementation Scheme 59, wherein the SPLAM is indisulan or a pharmaceutically acceptable salt thereof.

[0318] Implementation Scheme 61. In Implementation Scheme 61, the SPLAM described in Implementation Scheme 59, wherein the SPLAM is selected from the group consisting of: E7820, chloroquine sulfonamide, and tacisulan or a pharmaceutically acceptable salt thereof.

[0319] Implementation Scheme 62. In Implementation Scheme 62, a MAT2A inhibitor for treating cancer is provided, wherein the MAT2A inhibitor is administered simultaneously or sequentially with a topoisomerase inhibitor.

[0320] Implementation Scheme 63. In Implementation Scheme 63, the topoisomerase inhibitor described in Implementation Scheme 62, wherein the topoisomerase inhibitor is a type I topoisomerase inhibitor.

[0321] Implementation Scheme 64. In Implementation Scheme 64, the type I topoisomerase inhibitor described in Implementation Scheme 63, wherein the type I topoisomerase inhibitor is selected from the group consisting of 10-hydroxycamptothecin, irinotecan and topotecan, or pharmaceutically acceptable salts thereof.

[0322] Implementation Scheme 64A. In Implementation Scheme 64A, the use according to Implementation Scheme 63 is provided, wherein the type I topoisomerase inhibitor is selected from the group consisting of: hexylresorcinol, ethatecan, delutecan and belotecone, or pharmaceutically acceptable salts thereof.

[0323] Implementation Scheme 65. In Implementation Scheme 65, the topoisomerase inhibitor described in Implementation Scheme 62, wherein the topoisomerase inhibitor is a type II topoisomerase inhibitor.

[0324] Implementation Scheme 66. In Implementation Scheme 66, the use of the type II topoisomerase inhibitor described in Implementation Scheme 65, wherein the type II topoisomerase inhibitor is selected from the group consisting of: daunorubicin, doxorubicin and etoposide, or pharmaceutically acceptable salts thereof.

[0325] Implementation Scheme 67. In Implementation Scheme 67, the use of the MAT2A inhibitor described in any one of Implementation Schemes 56 to 66 is provided, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I); in X is either N or CH; R 3 C 1-6 Halogenated alkyl, halogenated or C3-6 cycloalkyl; R 2 For -NR 4 R 5 ; R 4 It is hydrogen or C 1-6 alkyl; R 5 For hydrogen, C 1-6 Alkyl or C 3-6 cycloalkyl; and R 1 It is a phenyl group, wherein the phenyl group is substituted by 0-2 halogen groups.

[0326] Implementation Scheme 68. In Implementation Scheme 68, the use of the MAT2A inhibitor described in Implementation Scheme 67 is provided, wherein the MAT2A inhibitor is a compound of formula (I) as defined in any one of Implementation Schemes 24 to 31 or a pharmaceutically acceptable salt thereof.

[0327] Implementation Scheme 69. In Implementation Scheme 69, the use of the MAT2A inhibitor described in any one of Implementation Schemes 56 to 68 is provided, wherein the MAT2A inhibitor is selected from the group consisting of compounds in Table 1 or pharmaceutically acceptable salts thereof.

[0328] Implementation Scheme 70. In Implementation Scheme 70, the use of the MAT2A inhibitor described in any one of Implementation Schemes 56 to 69 is provided, wherein the MAT2A inhibitor is compound A or a pharmaceutically acceptable salt thereof.

[0329] Implementation Scheme 71. In Implementation Scheme 71, the use of the MAT2A inhibitor described in any one of Implementation Schemes 56 to 69 is provided, wherein the MAT2A inhibitor is compound A1 or a pharmaceutically acceptable salt thereof.

[0330] Implementation Scheme 72. In Implementation Scheme 72, a use according to any one of Implementation Schemes 40 to 71 is provided, wherein the cancer is selected from the group consisting of: leukemia, glioma, lung cancer, esophageal cancer, MTAP-deficient pancreatic ductal adenocarcinoma (PDAC), melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, anal cancer, gastric cancer, colon cancer, colorectal cancer, soft tissue sarcoma, non-Hodgkin lymphoma, gastric cancer, esophageal gastric cancer, malignant peripheral nerve sheath tumor, mesothelioma, salivary gland tumor, urothelial carcinoma, gastrointestinal cancer, and sarcoma.

[0331] Implementation Scheme 73. In Implementation Scheme 73, the use according to any one of Implementation Schemes 40 to 72 is provided, wherein the cancer is a solid tumor or a hematologic cancer.

[0332] Implementation Scheme 74. In Implementation Scheme 74, the use according to any one of Implementation Schemes 40 to 73 is provided, wherein the cancer is a solid tumor.

[0333] Implementation Scheme 75. In Implementation Scheme 75, the use according to any one of Implementation Schemes 40 to 74 is provided, wherein the cancer is a solid malignant tumor.

[0334] Implementation Scheme 76. In Implementation Scheme 76, the use according to any one of Implementation Schemes 40 to 75 is provided, wherein the cancer is characterized by reduced or absent expression of the MTAP gene, absence of the MTAP gene, reduced function of the MTAP protein, reduced or absent level of the MTAP protein, accumulation of MTA, or a combination thereof.

[0335] Implementation Scheme 77. In Implementation Scheme 77, a method according to any one of Implementation Schemes 40 to 76 is provided, wherein the cancer is characterized by the deletion of the MTAP gene.

[0336] Implementation Scheme 78. In Implementation Scheme 78, a method is provided to inhibit tumor growth or slow the tumor growth rate in a subject with MTAP-deficient cancer, the method comprising administering a MAT2A inhibitor and a KIF18 inhibitor, SPLAM, or a topoisomerase inhibitor.

[0337] Implementation Scheme 79. In Implementation Scheme 79, the method according to Implementation Scheme 78 is provided, wherein tumor growth is measured by changes in tumor volume from a first time point to a second time point.

[0338] Implementation Scheme 80. In Implementation Scheme 80, the method according to Implementation Scheme 79 is provided, wherein the tumor volume at the second time point does not increase compared to the first time point.

[0339] Implementation Scheme 81. In Implementation Scheme 81, the method according to Implementation Scheme 80 is provided, wherein the tumor volume decreases between a first time point and a second time point.

[0340] Implementation Scheme 82. In Implementation Scheme 82, a method for treating cancer in a subject of need is provided, the method comprising administering to the subject a MAT2A inhibitor and a KIF18 inhibitor, SPLAM or a topoisomerase inhibitor, wherein the subject has previously undergone a cancer treatment regimen without the use of a MAT2A inhibitor.

[0341] Implementation Scheme 83. In Implementation Scheme 83, the method according to Implementation Scheme 82 is provided, wherein treatment with MAT2A inhibitors and KIF18 inhibitors, SPLAM or topoisomerase inhibitors reduces tumor growth rate compared to treatment with MAT2A inhibitors alone for a similar time period.

[0342] Implementation Scheme 84. In Implementation Scheme 84, a combination product is provided comprising a MAT2A inhibitor and an inhibitor selected from the group consisting of a KIF18 inhibitor, SPLAM and a topoisomerase inhibitor.

[0343] Implementation Scheme 85. In Implementation Scheme 85, a combination product is provided, comprising a first pharmaceutical composition and a second pharmaceutical composition, the first pharmaceutical composition comprising a therapeutically effective amount of a MAT2A inhibitor, and the second pharmaceutical composition comprising a therapeutically effective amount of an inhibitor selected from the group consisting of a KIF18 inhibitor, SPLAM, and a topoisomerase inhibitor.

[0344] Implementation Scheme 86. In Implementation Scheme 86, the MAT2A inhibitor described in Implementation Scheme 84 or 85 is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I); in X is either N or CH; R 3 C 1-6 Halogenated alkyl, halogenated or C 3-6 cycloalkyl; R 2 For -NR 4 R 5 ; R 4 It is hydrogen or C 1-6 alkyl; R 5 For hydrogen, C 1-6 Alkyl or C 3-6 cycloalkyl; and R 1 It is a phenyl group, wherein the phenyl group is substituted by 0-2 halogen groups.

[0345] Implementation Scheme 87. In Implementation Scheme 87, the MAT2A inhibitor described in Implementation Scheme 86 is a compound of formula (I) as defined in any one of Implementation Schemes 24 to 31, or a pharmaceutically acceptable salt thereof.

[0346] Implementation Scheme 88. In Implementation Scheme 88, the MAT2A inhibitor described in any one of Implementation Schemes 84 to 87 is selected from the group consisting of compounds in Table 1 or pharmaceutically acceptable salts thereof.

[0347] Example 89. In Example 89, the MAT2A inhibitor described in any one of Examples 84 to 88 is compound A or a pharmaceutically acceptable salt thereof.

[0348] Implementation Scheme 90. In Implementation Scheme 90, the MAT2A inhibitor described in any one of Implementation Schemes 84 to 88 is compound A1 or a pharmaceutically acceptable salt thereof.

[0349] Implementation Scheme 91. In Implementation Scheme 91, the KIF18 inhibitor described in any one of Implementation Schemes 84 to 90 is a KIF18A inhibitor.

[0350] Implementation Scheme 92. In Implementation Scheme 92, the KIF18A inhibitor described in Implementation Scheme 91 is, wherein the KIF18A inhibitor is sovexib (compound B) or a pharmaceutically acceptable salt thereof.

[0351] Implementation Scheme 93. In Implementation Scheme 93, the SPLAM of any one of Implementation Schemes 84 to 90, wherein the SPLAM is indisulan or a pharmaceutically acceptable salt thereof.

[0352] Implementation Scheme 94. In Implementation Scheme 94, the SPLAM of any one of Implementation Schemes 84 to 90, wherein the SPLAM is selected from the group consisting of: E7820, chloroquine sulfonamide and tacisulan or a pharmaceutically acceptable salt thereof.

[0353] Implementation Scheme 95. In Implementation Scheme 95, the topoisomerase inhibitor of any one of Implementation Schemes 84 to 90, wherein the topoisomerase inhibitor is a type I topoisomerase inhibitor.

[0354] Implementation Scheme 96. In Implementation Scheme 96, the type I topoisomerase inhibitor described in Implementation Scheme 95, wherein the type I topoisomerase inhibitor is selected from the group consisting of 10-hydroxycamptothecin, irinotecan and topotecan, or pharmaceutically acceptable salts thereof.

[0355] Example 96A. In Example 96A, the type I topoisomerase inhibitor described in Example 95 is selected from the group consisting of: hexylresorcinol, ethatecan, delutecan, and belotecone, or pharmaceutically acceptable salts thereof.

[0356] Implementation Scheme 96B. In Implementation Scheme 96B, the type I topoisomerase inhibitor described in Implementation Scheme 95 is irinotecan.

[0357] Implementation Scheme 97. In Implementation Scheme 97, the topoisomerase inhibitor of any one of Implementation Schemes 84 to 90, wherein the topoisomerase inhibitor is a type II topoisomerase inhibitor.

[0358] Implementation Scheme 98. In Implementation Scheme 98, the use of the type II topoisomerase inhibitor described in Implementation Scheme 97, wherein the type II topoisomerase inhibitor is selected from the group consisting of: daunorubicin, doxorubicin and etoposide, or pharmaceutically acceptable salts thereof.

[0359] Implementation Scheme 99. In Implementation Scheme 99, a method for treating cancer in a subject of need is provided, the method comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and administering to the subject a therapeutically effective amount of an antibody-drug conjugate (ADC) comprising a topoisomerase inhibitor, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0360] Implementation Scheme 100. In Implementation Scheme 100, the ADC containing the topoisomerase inhibitor is a type I topoisomerase inhibitor.

[0361] Implementation Scheme 101. Implementation Scheme 101, the ADC containing a topoisomerase inhibitor is trastuzumab.

[0362] Implementation scheme 102. Implementation scheme 102, the ADC containing the topoisomerase inhibitor is AZD8205.

[0363] Implementation scheme 103. Implementation scheme 103, wherein the ADC containing the topoisomerase inhibitor is DS-1062.

[0364] Implementation Scheme 104. In Implementation Scheme 104, the MAT2A inhibitor described in any one of Implementation Schemes 99 to 103 is selected from the group consisting of the compounds in Table 1 or their pharmaceutically acceptable salts.

[0365] Implementation Scheme 105. In Implementation Scheme 105, the MAT2A inhibitor described in any one of Implementation Schemes 99 to 104 is compound A or a pharmaceutically acceptable salt thereof.

[0366] Implementation Scheme 106. In Implementation Scheme 106, the MAT2A inhibitor described in any one of Implementation Schemes 99 to 104 is compound A1 or a pharmaceutically acceptable salt thereof.

[0367] Implementation Scheme 107. In Implementation Scheme 107, a method according to any one of Implementation Schemes 99 to 106 is provided, wherein the cancer is selected from the group consisting of: leukemia, glioma, lung cancer, esophageal cancer, MTAP-deficient pancreatic ductal adenocarcinoma (PDAC), melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, anal cancer, gastric cancer, colon cancer, colorectal cancer, soft tissue sarcoma, non-Hodgkin lymphoma, gastric cancer, esophageal gastric cancer, malignant peripheral nerve sheath tumor, mesothelioma, salivary gland tumor, urothelial carcinoma, gastrointestinal cancer, and sarcoma.

[0368] Implementation Scheme 108. In Implementation Scheme 108, a method according to any one of Implementation Schemes 99 to 107 is provided, wherein the cancer is a solid tumor or a hematologic cancer.

[0369] Implementation Scheme 109. In Implementation Scheme 109, a method according to any one of Implementation Schemes 99 to 108 is provided, wherein the cancer is a solid tumor.

[0370] Implementation Scheme 110. In Implementation Scheme 110, a method according to any one of Implementation Schemes 99 to 109 is provided, wherein the cancer is a solid malignant tumor.

[0371] Implementation Scheme 111. In Implementation Scheme 111, a method according to any one of Implementation Schemes 99 to 110 is provided, wherein the cancer is characterized by reduced or absent MTAP gene expression, absence of the MTAP gene, reduced function of the MTAP protein, reduced or absent level of the MTAP protein, accumulation of MTA, or a combination thereof.

[0372] Implementation Scheme 112. In Implementation Scheme 112, a method according to any one of Implementation Schemes 1 to 111 is provided, wherein the cancer is characterized by the deletion of the MTAP gene.

[0373] For the above embodiments, when referring to previous embodiments, the reference will include embodiments with letter designations or combinations thereof. For example, a reference to embodiments 40 to 50 will include embodiments 40, 41, 42, 43, 44, 45, 46, 47, 48, 48A, 49, and 50. Part B - Additional Non-Limiting Exemplary Embodiments 1 to 40

[0374] Implementation Scheme 1. In Implementation Scheme 1, a method for treating cancer in a subject in need is provided, the method comprising administering a MAT2A inhibitor to the subject and administering a therapeutically effective amount of a topoisomerase inhibitor to the subject, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I); in X is either N or CH; R 3 C 1-6 Halogenated alkyl, halogenated or C 3-6 cycloalkyl; R 2 For -NR 4 R 5 ; R 4 It is hydrogen or C 1-6 alkyl; R 5 For hydrogen, C 1-6 Alkyl or C 3-6 cycloalkyl; and R 1 It is a phenyl group, wherein the phenyl group is substituted by 0-2 halogen groups.

[0375] Implementation Scheme 2. In Implementation Scheme 2, the method according to Implementation Scheme 1 is provided, wherein the topoisomerase inhibitor is a type I topoisomerase inhibitor.

[0376] Implementation Scheme 3. In Implementation Scheme 3, the method according to Implementation Scheme 2 is provided, wherein the type I topoisomerase inhibitor is selected from the group consisting of 10-hydroxycamptothecin, irinotecan and topotecan, or pharmaceutically acceptable salts thereof.

[0377] Implementation Scheme 4. In Implementation Scheme 4, the method according to Implementation Scheme 1 is provided, wherein the topoisomerase inhibitor is a type II topoisomerase inhibitor.

[0378] Implementation Scheme 5. In Implementation Scheme 5, the method according to Implementation Scheme 4 is provided, wherein the type II topoisomerase inhibitor is selected from the group consisting of: daunorubicin, doxorubicin and etoposide, or pharmaceutically acceptable salts thereof.

[0379] Implementation Scheme 6. In Implementation Scheme 6, a method according to any one of Implementation Schemes 1 to 5 is provided, wherein the MAT2A inhibitor and the topoisomerase inhibitor are administered simultaneously or sequentially.

[0380] Implementation Scheme 7. In Implementation Scheme 7, a method for treating cancer in a subject of need is provided, the method comprising administering a MAT2A inhibitor to the subject and administering to the subject a therapeutically effective amount of a splice inhibitor sulfonamide (SPLAM), wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I); in X is either N or CH; R 3 C 1-6 Halogenated alkyl, halogenated or C 3-6 cycloalkyl; R 2 For -NR 4 R 5 ; R 4 It is hydrogen or C 1-6 alkyl; R 5 For hydrogen, C 1-6 Alkyl or C 3-6 cycloalkyl; and R 1 It is a phenyl group, wherein the phenyl group is substituted by 0-2 halogen groups.

[0381] Implementation Scheme 8. In Implementation Scheme 8, the method according to Implementation Scheme 7 is provided, wherein the SPLAM is indisulan or a pharmaceutically acceptable salt thereof.

[0382] Implementation Scheme 9. In Implementation Scheme 9, the method according to Implementation Scheme 7 is provided, wherein the SPLAM is selected from the group consisting of: E7820, chloroquine sulfonamide, and tacisulan or a pharmaceutically acceptable salt thereof.

[0383] Implementation Scheme 10. In Implementation Scheme 10, a method according to any one of Implementation Schemes 7 to 9 is provided, wherein the MAT2A inhibitor is administered simultaneously or sequentially with SPLAM.

[0384] Implementation Scheme 11. In Implementation Scheme 11, a method for treating cancer in a subject of need is provided, the method comprising administering a MAT2A inhibitor to the subject and administering a therapeutically effective amount of a KIF18 inhibitor to the subject, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I); in X is either N or CH; R 3 C 1-6 Halogenated alkyl, halogenated or C 3-6 cycloalkyl; R 2 For -NR 4 R 5 ; R 4 It is hydrogen or C 1-6 alkyl; R 5 For hydrogen, C 1-6Alkyl or C 3-6 cycloalkyl; and R 1 It is a phenyl group, wherein the phenyl group is substituted by 0-2 halogen groups.

[0385] Implementation Scheme 12. In Implementation Scheme 12, the method according to Implementation Scheme 11 is provided, wherein the KIF18 inhibitor is a KIF18A inhibitor.

[0386] Implementation Scheme 13. In Implementation Scheme 13, the method according to Implementation Scheme 12 is provided, wherein the KIF18A inhibitor is sovexib or a pharmaceutically acceptable salt thereof.

[0387] Implementation Scheme 14. In Implementation Scheme 14, a method according to any one of Implementation Schemes 11B to 13B is provided, wherein the MAT2A inhibitor and the KIF18 inhibitor are administered simultaneously or sequentially.

[0388] Implementation Scheme 15. In Implementation Scheme 15, a method according to any one of Implementation Schemes 1 to 14 is provided, wherein X is N.

[0389] Implementation Scheme 16. In Implementation Scheme 16, a method according to any one of Implementation Schemes 1 to 14 is provided, wherein X is CH.

[0390] Implementation Scheme 17. In Implementation Scheme 17, a method according to any one of Implementation Schemes 1 to 16 is provided, wherein R 4 It is hydrogen and R 5 It is hydrogen or C 1-3 alkyl.

[0391] Implementation Scheme 18. In Implementation Scheme 18, a method according to any one of Implementation Schemes 1 to 17 is provided, wherein R 5 It can be hydrogen or methyl.

[0392] Implementation Scheme 19. In Implementation Scheme 19, a method according to any one of Implementation Schemes 1 to 18 is provided, wherein R 1 It is a phenyl group that has been substituted with chlorine.

[0393] Implementation Scheme 20. In Implementation Scheme 20, a method according to any one of Implementation Schemes 1 to 19 is provided, wherein R 3 C 1-3 Halogenated alkyl or halogenated.

[0394] Implementation Scheme 21. In Implementation Scheme 21, a method according to any one of Implementation Schemes 1 to 20 is provided, wherein R 3 It is trifluoromethyl or chlorine.

[0395] Implementation Scheme 22. In Implementation Scheme 22, a method according to any one of Implementation Schemes 1 to 20 is provided, wherein R 3 It is cyclopropyl.

[0396] Implementation Scheme 23. In Implementation Scheme 23, a method according to any one of Implementation Schemes 1 to 22 is provided, wherein the MAT2A inhibitor is selected from the group consisting of compounds in Table 1 or pharmaceutically acceptable salts thereof.

[0397] Implementation Scheme 24. In Implementation Scheme 24, a method according to any one of Implementation Schemes 1 to 23 is provided, wherein the MAT2A inhibitor is compound A:

[0398] Compound A

[0399] Or its pharmaceutically acceptable salt.

[0400] Implementation Scheme 25. In Implementation Scheme 25, a method according to any one of Implementation Schemes 1 to 23 is provided, wherein the MAT2A inhibitor is compound A1:

[0401] Compound A1

[0402] Or its pharmaceutically acceptable salt.

[0403] Implementation Scheme 26. In Implementation Scheme 26, a method according to any one of Implementation Schemes 1 to 25 is provided, wherein the cancer is selected from the group consisting of: leukemia, glioma, lung cancer, esophageal cancer, MTAP-deficient pancreatic ductal adenocarcinoma (PDAC), melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, anal cancer, gastric cancer, colon cancer, colorectal cancer, soft tissue sarcoma, non-Hodgkin lymphoma, gastric cancer, esophageal cancer, esophageal cancer, malignant peripheral nerve sheath tumor, mesothelioma, salivary gland tumor, urothelial carcinoma, gastrointestinal cancer, and sarcoma.

[0404] Implementation Scheme 27. In Implementation Scheme 27, a method according to any one of Implementation Schemes 1 to 26 is provided, wherein the cancer is a solid tumor or a hematologic cancer.

[0405] Implementation Scheme 28. In Implementation Scheme 28, a method according to any one of Implementation Schemes 1 to 27 is provided, wherein the cancer is a solid malignant tumor.

[0406] Implementation Scheme 29. In Implementation Scheme 29, a method according to any one of Implementation Schemes 1 to 28 is provided, wherein the cancer is characterized by reduced or absent expression of the MTAP gene, absence of the MTAP gene, reduced function of the MTAP protein, reduced or absent level of the MTAP protein, accumulation of MTA, or a combination thereof.

[0407] Implementation Scheme 30. In Implementation Scheme 30, the use of a MAT2A inhibitor in the manufacture of a medicament for the treatment of cancer is provided, wherein the MAT2A inhibitor is administered together with a topoisomerase inhibitor.

[0408] Implementation Scheme 31. In Implementation Scheme 31, the use of a MAT2A inhibitor in the manufacture of a medicament for the treatment of cancer is provided, wherein the MAT2A inhibitor is administered in combination with SPLAM.

[0409] Implementation Scheme 32. In Implementation Scheme 32, the use of a MAT2A inhibitor in the manufacture of a medicament for the treatment of cancer is provided, wherein the MAT2A inhibitor is administered in combination with a KIF18 inhibitor.

[0410] Implementation Scheme 33. In Implementation Scheme 33, the use described in any one of Implementation Schemes 30 to 32 is provided, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0411] Implementation Scheme 34. In Implementation Scheme 34, the use described in Implementation Scheme 33B is provided, wherein the MAT2A inhibitor is compound A or a pharmaceutically acceptable salt thereof, or compound A1 or a pharmaceutically acceptable salt thereof.

[0412] Implementation Scheme 35. In Implementation Scheme 35, a method is provided to inhibit tumor growth or slow the tumor growth rate in a subject with MTAP-deficient cancer, the method comprising administering a MAT2A inhibitor and a topoisomerase inhibitor, SPLAM, or a KIF18 inhibitor.

[0413] Implementation Scheme 36. In Implementation Scheme 36, the method according to Implementation Scheme 35 is provided, wherein tumor growth is measured by changes in tumor volume from a first time point to a second time point.

[0414] Implementation Scheme 37. In Implementation Scheme 37, the method according to Implementation Scheme 36 is provided, wherein the tumor volume at the second time point does not increase compared to the first time point.

[0415] Implementation Scheme 38. In Implementation Scheme 38, the method according to Implementation Scheme 37 is provided, wherein the tumor volume decreases between a first time point and a second time point.

[0416] Implementation Scheme 39. In Implementation Scheme 39, a method for treating cancer in a subject of need is provided, the method comprising administering to the subject a MAT2A inhibitor and a topoisomerase inhibitor, SPLAM or KIF18 inhibitor, wherein the subject has previously undergone a cancer treatment regimen without the use of a MAT2A inhibitor.

[0417] Implementation Scheme 40. In Implementation Scheme 40, the method according to Implementation Scheme 39 is provided, wherein treatment with a MAT2A inhibitor and a topoisomerase inhibitor, SPLAM or KIF18 inhibitor reduces the tumor growth rate compared with treatment with a MAT2A inhibitor alone for a similar time period.

[0418] Those skilled in the art will recognize or be able to determine numerous equivalents of the particular procedures, embodiments, claims, and examples described herein using only conventional experiments. Such equivalents are considered to be within the scope of this disclosure and are covered by the appended claims. For example, it should be understood that modifications to reaction conditions, including but not limited to reaction time, reaction size / volume, and experimental reagents such as solvents, catalysts, pressure, atmospheric conditions (e.g., nitrogen atmosphere), and reducing / oxidizing agents, using art-recognized alternatives and employing only conventional experiments, are all within the scope of this application.

[0419] It should be understood that wherever values ​​and ranges are provided herein, all values ​​and ranges covered by such values ​​and ranges are intended to be covered within the scope of this disclosure. Furthermore, all values ​​falling within these ranges, as well as the upper or lower limits of the ranges of values, are also covered by this application.

[0420] The following examples further illustrate various aspects of this disclosure. However, they in no way constitute a limitation on the teachings of this disclosure.

[0421] Example

[0422] The compounds and methods disclosed herein are further illustrated by the following examples, which should not be considered as further limitations. Unless otherwise indicated, the practice of this disclosure will utilize conventional techniques of organic synthesis, cell biology, cell culture, and molecular biology, which are within the scope of the art.

[0423] Example 1: In vitro synergistic growth inhibition of an equimolar combination of MAT2A inhibitor and KIF18A inhibitor

[0424] Materials and methods

[0425] In a group of 5 MTAPTen-day proliferation assays were performed in defective and two wild-type non-small cell lung cancer and bladder cancer cell lines. Optimal cell seeding for all cell lines was determined by evaluating growth at different seeding densities in 384-well plates to identify conditions allowing for 10 days of proliferation. Cells were then seeded at the optimal seeding density and treated with 20-point, two-fold dilution series of compound A, sovexib (“Compound B”), or an equimolar combination of Compound A and Compound B. The concentrations of Compound A and Compound B, tested alone or in combination, ranged from 0.038 nM to 20,000 nM. One plate of cells was harvested upon addition of the compounds to quantify the cell number at the start of the combination (T0). For cell quantification, harvested cells were lysed with Promega CellTiter-Glo (CTG) reagent according to the manufacturer’s protocol, and chemiluminescence signals were detected on a Synergy Neo plate reader. CTG estimated cell number by detecting cellular ATP levels. Cells were cultured with the drug combination at 37°C and 5% CO2 for 10 days. Cells were then lysed with CTG, and chemiluminescence signals were measured.

[0426] analyze

[0427] The CTG values ​​obtained after 10 days of treatment were subtracted from the background values ​​and expressed as a percentage of the T0 value. The data were plotted against compound concentrations and fitted with a four-parameter equation to generate dose-response curves for each single agent and equimolar combinations. Growth ICx (gICx) values ​​ranging from gIC30 to gIC100 were interpolated from the fitted curves. Synergistic growth inhibition was assessed by determining the combination index (CI) values ​​at various points during titration using a non-restrictive equation [Chou, 1983; Chou, 1981], where A equals the gIC value of compound A, B equals the gICx value of compound B, and A+B or B+A equals the gICx value of the equimolar combination. A combination index value less than 1.0 was considered synergistic.

[0428]

[0429] result

[0430] In 5 / 5 MTAP In the defective cell lines, synergistic growth inhibition of the combination of compound A and compound B was observed at multiple gICx points during the combinatorial titration process (Tables 3A-3E). Combination synergy was also observed in half of the wild-type cell lines (Tables 4A-4B). Although considered synergistic, the combination index value in the wild-type TCCSUP cell lines was ≥0.7. Compared to the wild-type cell lines, MTAPDefective cell lines showed higher sensitivity to combinations, with combination index values ​​as low as 0.01.

[0431] Tables 3A-3E: Synergistic growth inhibition of five MTAP-deficient cell lines by equimolar combinations of compounds A and B.

[0432] Using a non-repulsive combinatorial index equation, the synergistic effect of equimolar combinations of compounds A and B was measured within the gICx range of gIC30-gIC100. The combinatorial index values ​​are shown as: NCI-H838 (3A), SW900 (3B), RT112 / 84 (3C), UMUC5 (3D), and UMUC11 (3E).

[0433] Table 3A: NCI-H838

[0434] Table 3B: SW900

[0435] Table 3C: RT112 / 84

[0436] Table 3D: UMUC5

[0437] Table 3E: UMUC11

[0438] Tables 4A-4B: Synergistic growth inhibition of two wild-type cell lines by equimolar combinations of compounds A and B.

[0439] The synergistic effect of equimolar combinations of compounds A and B was measured using a non-repulsive combinatorial index equation within the gICx range of gIC30-gIC100. The combinatorial index values ​​are shown as NCI-H520 (4A) and TCCSUP (4B).

[0440] Table 4A: NCI-H520

[0441] Table 4B: TCCSUP

[0442] Example 2: In vitro synergistic growth inhibition of a combination of KIF18A inhibitor and a fixed concentration of MAT2A inhibitor

[0443] Materials and methods

[0444] In a group of 5 MTAP Ten-day proliferation assays were performed in defective and two wild-type non-small cell lung cancer and bladder cancer cell lines. Optimal cell seeding for all cell lines was determined by evaluating growth at different seeding densities in 384-well plates to identify conditions allowing for 10 days of proliferation. Cells were then seeded at the optimal seeding density and treated with a combination of compound B in a 20-point, two-fold dilution series with a fixed concentration of 1000 nM of compound A. Single-drug titrations of compounds A and B were included for comparison. The concentrations of compounds A and B, tested alone or in combination, ranged from 0.038 nM to 20,000 nM. One plate of cells was harvested upon addition of the compounds to quantify the cell count at the start of the combination (T0). For cell quantification, the harvested cells were lysed with Promega CellTiter-Glo (CTG) reagent according to the manufacturer's protocol, and chemiluminescence signals were detected as described in Example 1 above. Cells and drug combinations were cultured at 37°C and 5% CO2 for 10 days. Cells were then lysed with CTG, and chemiluminescence signals were measured.

[0445] analyze

[0446] The CTG values ​​obtained after 10 days of treatment were subtracted from the background values ​​and expressed as a percentage of the T0 value set at 100%. The data were plotted against compound concentrations and fitted with a four-parameter equation to generate dose-response curves for each single agent. The percentage of inhibition for each combination and single drug concentration was determined compared to the DMSO control. In addition, the overall net growth or death of cells was calculated and reported as a growth-death index (GDI) value ranging from -100 to 100, with the midpoint (0) representing the number of cells at the start of the combination (T0).

[0447] Synergistic growth inhibition was assessed at each point using the Bliss independence model for the titration of compound B in combination with 1000 nM compound A. The observed growth inhibition at each point was compared with the inhibition predicted based on the additive activity of each single agent. Predicted inhibition was calculated using (Ea + Eb) - (Ea) The Bliss score is calculated using Eb), where E equals the effect (inhibition) of each individual agent (a and b). A difference greater than 10% between the observed and predicted percentage of inhibition is considered synergistic. The Bliss score is calculated only for combinations causing >20% growth inhibition (GDI value <80). Because the Bliss score is based on growth inhibition, scores within the cytotoxic dose range may not accurately reflect synergistic cytotoxicity.

[0448] result

[0449] In 5 / 5 MTAP In defective cell lines, synergistic growth inhibition of the combination of compound B and 1000 nM compound A was observed at multiple titration points, as assessed by Bliss independence (Tables 5A-5E). Due to the drastic changes in the dose-response curves observed with single-agent compound B, the window for observing synergistic effects in this assay design is narrow for some cell lines. For example, 5000 nM of single-agent compound B was sufficient to maximally inhibit cell growth in the UMUC5 cell line (…). Figure 1A-3 Therefore, the dose range for observed synergistic effects of the combination in UMUC5 is below 5000 nM of compound B. Although the window is limited, it is within 5 / 5 of the range. MTAP In defective cell lines, synergistic effects of the combination were observed at doses below those required for maximum single-agent inhibition. In these cell lines, synergistic growth inhibition was observed in the combination of compound B and 1000 nM compound A over a wide dose range.

[0450] Enhanced growth inhibition by the combination was also observed in half of the wild-type cell lines within a limited dose range of 39–156 nM for compound B (Tables 6A–6B). Although synergistic effects in this cell line were calculated using Bliss independence, the change in potency in the dose-response curves was not as pronounced as in… MTAP Not as large as those observed in defective cell lines. Figures 1A-1 to 1A-5 and Figures 1B-1 to 1B-2 In this study, growth inhibition was expressed as a percentage of the DMSO control after titration with a single agent of compound B. Five [specific compounds] were generated using four-parameter fitting. MTAP Defective cell lines ( Figures 1A-1 to 1A-5 ) and two wild-type cell lines ( Figure 1B-1 and 1B-2 The dose-response curve of ).

[0451] Tables 5A-5E: Synergistic growth inhibition of five MTAP-deficient cell lines by the combination of compound B and 1000 nM compound A.

[0452] The synergistic effect of compound B in the titration of 1000 nM compound A was measured using Bliss independence analysis. A Bliss score >10 was considered "...". The terms “” indicate that the combination is considered to have a synergistic effect. The Bliss score applies to combinations that exhibit a growth inhibition rate greater than 20% (GDI < 80). Synergistic effects reported using Bliss may be underrepresented in the cytotoxic dose range. Growth and death index values ​​for combination titration and for each single agent compound A (Cmpd A) and compound B (Cmpd B) are shown, ranging from -100 to 100, with the midpoint (0) representing the cell number at the start of the combination (T0). Bliss and GDI values ​​for five MTAP-deficient cell lines are shown: NCI-H838 (5A), SW900 (5B), RT112 / 84 (5C), UMUC5 (5D), and UMUC11 (5E).

[0453] Table 5A: NCI-H838

[0454] Table 5B: SW900

[0455] Table 5C: RT112 / 84

[0456] Table 5D: UMUC5

[0457] Table 5E: UMUC11

[0458] Tables 6A-6B: Synergistic growth inhibition of two wild-type cell lines by the combination of compound B and 1000 nM compound A.

[0459] The synergistic effect of compound B in the titration of 1000 nM compound A was measured using Bliss independence analysis. A Bliss score >10 was considered "...". The terms "+" indicate that the combination is considered to have a synergistic effect. Bliss scores were calculated only for combinations exhibiting a growth inhibition rate greater than 20%. Synergistic effects reported using Bliss may be underrepresented across the cytotoxic dose range. Growth and death index values ​​for combination titration and for each individual agent, compound A (Cmpd A) and compound B (Cmpd B), are shown, ranging from -100 to 100, with the midpoint (0) representing the cell number at the start of the combination (T0). Bliss and GDI values ​​for five MTAP-deficient cell lines are shown: NCI-H520 (6A), TCCSUP (6B).

[0460] Table 6A: NCI-H520

[0461] Table 6B: TCCSUP

[0462] Example 3: Proliferation screening to identify the combined benefits of MAT2A inhibition and indisulan

[0463] Materials and methods

[0464] Ten-day proliferation screening was performed on 15 pancreatic cancer, non-small cell lung cancer, and bladder cancer cell lines: 10 MTAP Ineffective cell lines and 5 MTAP Wild-type cell lines. Optimal seeding for all cell lines was determined by evaluating growth at different seeding densities in 384-well plates to identify conditions allowing for 10 days of proliferation. Cells were then seeded at the optimal seeding density in the presence of either 20–150 nM compound A or DMSO mediator controls. Cells were cultured at 37°C and 5% CO2 for 4 days to allow the pretreatment compounds to bind to the target compounds. While maintaining the pretreatment conditions, cells were also treated with an 11-point, triple-titration series of compounds from a chemiluminescence library of 424 compounds. The concentrations of the combined compounds ranged from 0.2 nM to 14,679 nM. One plate of cells was harvested upon addition of the combined compounds to quantify the cell count at the start of the combination (T0). For cell quantification, harvested cells were lysed with CellTiter-Glo (CTG) (Promega) reagent according to the manufacturer's protocol, and chemiluminescence signals were detected on a Synergy Neo plate reader (ThermoFisher, Serial No. 140715A). CTG estimates cell number by detecting cellular ATP levels. Cells were incubated with the drug combination at 37°C and 5% CO2 for 6 days, for a total of 10 days including pretreatment. Cells were then lysed with CTG and chemiluminescent signals were measured. The CTG values ​​obtained after 10 days of treatment were subtracted from the background values ​​and expressed as a percentage of the T0 value, and plotted as a compound concentration graph. Data were fitted with a four-parameter equation to generate concentration-response curves. For each combination titration, the growth IC50 values ​​and maximum growth inhibition of cells pretreated with DMSO and compound A were compared. Combination hits were based on observations of a growth IC50 change >2-fold and / or a growth inhibition reduction greater than 20%.

[0465] result

[0466] In 10 MTAP Ineffective cell lines and 5 MTAP In wild-type cell lines, 424 combinations of compounds with compound A were tested. Of these compounds, indisulan was identified as the most consistent combination hit in the screening, ranking in the top 7 out of 10. MTAP In the defective cell lines and 1 / 5 of the wild-type cell lines, the growth IC50 changed ≥2-fold after pretreatment with compound A (Table 7).

[0467] Table 7: Growth IC50 of 10 cancer cell lines pretreated with and without compound A pretreatment 50 value

[0468] The growth IC of indisulan and compound A is shown. 50 And the multiple change value. In 7 / 10 MTAP Growth IC was observed in defective cell lines and 1 / 5 of wild-type cell lines. 50 The change in multiple is ≥ 2 times (in " The NCI-H838 cell line was tested at two concentrations of compound A (20 nM and 150 nM). Under both conditions, the growth IC50 was [value missing]. 50 The changes in the values ​​were all ≥2 times.

[0469]

[0470] Example 4: In vitro synergistic growth inhibition of equimolar combinations of MAT2A inhibitors and four structurally different splicing inhibitors, sulfonamides.

[0471] Materials and methods

[0472] In a group of 5 MTAP Ten-day proliferation assays were performed in defective and two wild-type non-small cell lung cancer and bladder cancer cell lines. Optimal cell seeding for all cell lines was determined by evaluating growth at different seeding densities in 384-well plates to identify conditions allowing for 10 days of proliferation. Cells were then seeded at the optimal seeding density and treated with 20-point, two-fold dilution series of compound A, the splicing inhibitor sulfonamide (SPLAM), or equimolar combinations of compound A and SPLAM. The tested combinations included combinations of compound A with four structurally different SPLAMs. The concentrations of compound A and each SPLAM tested alone or in combination ranged from 0.04 nM to 20,000 nM. One plate of cells was harvested upon addition of the compound to quantify the cell number at the start of the combination (T0). For cell quantification, harvested cells were lysed with Promega CellTiter-Glo (CTG) reagent according to the manufacturer's protocol, and chemiluminescent signals were detected on a Synergy Neo plate reader. CTG estimated cell number by detecting cellular ATP levels. Cells and the drug combination were cultured at 37°C and 5% CO2 for 10 days. Cells were then lysed using CTG and chemiluminescence signals were measured.

[0473] analyze

[0474] The CTG values ​​obtained after 10 days of treatment were subtracted from the background values ​​and expressed as a percentage of the T0 value. The data were plotted against compound concentrations and fitted with a four-parameter equation to generate dose-response curves for each single agent and equimolar combinations. Growth ICx (gICx) values ​​ranging from gIC30 to gIC100 were interpolated from the fitted curves. Synergistic growth inhibition was assessed by determining the combination index (CI) values ​​at various points during titration using the following non-restrictive equations [Chou, 1983; Chou, 1981], where A equals the gICx value of compound A, B equals the gICx value of SPLAM, and A+B or B+A equals the gICx value of the equimolar combination. A combination index value less than 1.0 was considered synergistic.

[0475]

[0476] result

[0477] In a group MTAP In the defective cell lines, synergistic growth inhibition of compound A in combination with each of the four different SPLAMs was observed at multiple gICx points during the combinatorial titration process (Tables 8A-8E, 9A-9E, 10A-10E, 11A-11E, and 12A-12E). Combinatorial synergy was also observed in both wild-type cell lines, particularly with the more potent SPLAMs indisulan and E7820 (Tables 8F-8G, 9F-9G, 10F-10G, and 11F-11G). Although considered synergistic, the combinatorial index values ​​in the wild-type cell lines were higher than those in the 8A-8E-9E-10E-11E-12E-12E-12E-12E-12E-13E-14E-15E-12E-13E-14E-15E-12E-13E-14E-15E-16E-17E-12E-13E-14E-15E-16E-17E-18E-19 ...6E-17E-18E-19E-12E-13E-16E-17E-18E-19E-12E-13E-16E-17E-18E-19E-12E-13E-16E-17E-18E-19E-12E-1 MTAP Values ​​observed in defective cell lines. Compared to wild-type cell lines, MTAP Defective cell lines showed higher sensitivity to the combination and greater growth inhibition.

[0478] Table 8A-8G: Synergistic growth inhibition of equimolar combinations of compound A and indisulan on five MTAP-deficient and two wild-type cell lines.

[0479] The synergistic effect of equimolar combinations of compound A and indisulan was measured using a non-exclusive combinatorial index equation within the gICx range of gIC30-gIC100. Combinatorial index values ​​for MTAP-deficient cell lines (8A-8E) and wild-type cell lines (8F-8G) are shown.

[0480] Table 8A: NCI-H838

[0481] Table 8B: SW900

[0482] Table 8C: RT112 / 84

[0483] Table 8D: UMUC5

[0484] Table 8E: UMUC11

[0485] Table 8F: NCI-H520

[0486] Table 8G: TCCSUP

[0487] Tables 9A-9G: Synergistic growth inhibition of equimolar combinations of compound A and E7820 on five MTAP-deficient and two wild-type cell lines.

[0488] The synergistic effect of equimolar combinations of compounds A and E7820 was measured using a non-exclusive combinatorial index equation within the gICx range of gIC30-gIC100. Combinatorial index values ​​for MTAP-deficient cell lines (9A-9E) and wild-type cell lines (9F-9G) are shown.

[0489] Table 9A: NCI-H838

[0490] Table 9B: SW900

[0491] Table 9C: RT112 / 84

[0492] Table 9D: UMUC5

[0493] Table 9E: UMUC11

[0494] Table 9F: NCI-H520

[0495] Table 9G: TCCSUP

[0496] Table 10A-10G: Synergistic growth inhibition of equimolar combinations of compound A and chloroquine sulfonamide on five MTAP-deficient and two wild-type cell lines.

[0497] The synergistic effect of equimolar combinations of compound A and chloroquine sulfonamide was measured using a non-exclusive combinatorial index equation within the gICx range of gIC30-gIC100. Combination index values ​​for MTAP-deficient cell lines (10A-10E) and wild-type cell lines (10F-10G) are shown.

[0498] Table 10A: NCI-H838

[0499] Table 10B: SW900

[0500] Table 10C: RT112 / 84

[0501] Table 10D: UMUC5

[0502] Table 10E: UMUC11

[0503] Table 10F: NCI-H520

[0504] Table 10G: TCCSUP

[0505] Table 11A-11G: Synergistic growth inhibition of equimolar combinations of compound A and tacisulan on five MTAP-deficient and two wild-type cell lines.

[0506] The synergistic effect of equimolar combinations of compound A and tacisulan was measured using a non-exclusive combinatorial index equation within the gICx range of gIC30-gIC100. Combinatorial index values ​​for MTAP-deficient cell lines (11A-11E) and wild-type cell lines (11F-11G) are shown.

[0507] Table 11A: NCI-H838

[0508] Table 11B: SW900

[0509] Table 11C: RT112 / 84

[0510] Table 11D: UMUC5

[0511] Table 11E: UMUC11

[0512] Table 11F: NCI-H520

[0513] Table 11G: TCCSUP

[0514] Tables 12A-12E: Synergistic growth inhibition of five MTAP-deficient cell lines by the combination of indisulan and 1000 nM compound A

[0515] The synergistic effect of indisulan in the titration of 1000 nM compound A was measured by Bliss independence analysis. A Bliss score >10 was considered "...". “” indicates and is considered to have a synergistic effect. The Bliss score applies to combinations that exhibit a growth inhibition rate greater than 20% (GDI < 80). Synergistic effects reported using Bliss may be underrepresented in the cytotoxic dose range. The growth and death index values ​​for combination titration and each single agent compound A (Cmpd A) and Indisulan are shown, ranging from -100 to 100, with the midpoint (0) being the cell number at the start of the combination (T0). Scores less than 0 are indicated by “”. “” indicates and represents cytotoxic response. Bliss and GDI values ​​for five MTAP-deficient cell lines are shown: NCI-H838 (12A), SW900 (12B), RT112 / 84 (12C), UMUC5 (12D), and UMUC11 (12E).

[0516] Table 12A: NCI-H838

[0517] Table 12B: SW900

[0518] Table 12C: RT112 / 84

[0519] Table 12D: UMUC5

[0520] Table 12E: UMUC11

[0521] Example 5: In vitro synergistic growth inhibition of a combination of four splicing inhibitors, sulfonamides, and a fixed concentration of MAT2A inhibitor.

[0522] Materials and methods

[0523] In a group of 5 MTAP Ten-day proliferation assays were performed in defective and two wild-type non-small cell lung cancer and bladder cancer cell lines. Optimal cell seeding for all cell lines was determined by evaluating growth at different seeding densities in 384-well plates to identify conditions allowing for 10 days of proliferation. Cells were then seeded at the optimal seeding density and treated with each of four structurally different SPLAMs in a 20-point, 2-fold dilution series, combined with a fixed concentration of 1000 nM of compound A. Single drug titrations of compound A and each SPLAM were included for comparison. The concentration range of each SPLAM tested alone or in combination was from 0.04 nM to 20,000 nM. One plate of cells was harvested upon addition of the compound to quantify the cell count at the start of the combination (T0). For cell quantification, the harvested cells were lysed with Promega CellTiter-Glo (CTG) reagent according to the manufacturer's protocol, and the chemiluminescence signal was detected as described in Example 1 above. Cells and drug combinations were cultured at 37°C and 5% CO2 for 10 days. Cells were then lysed with CTG, and the chemiluminescence signal was measured.

[0524] analyze

[0525] The CTG values ​​obtained after 10 days of treatment were subtracted from the background values ​​and expressed as a percentage of the T0 value set at 100%. The data were plotted against compound concentrations and fitted with a four-parameter equation to generate dose-response curves for each single agent. The percentage of inhibition for each combination and single drug concentration was determined compared to the DMSO control. In addition, the overall net growth or death of cells was calculated and reported as a growth-death index (GDI) value ranging from -100 to 100, with the midpoint (0) representing the number of cells at the start of the combination (T0).

[0526] Synergistic growth inhibition was assessed at each point using the Bliss independence model for the SPLAM titration combined with 1000 nM compound A. The observed growth inhibition at each point was compared with the predicted inhibition based on the additive activity of each single agent dose. Predicted inhibition was calculated using (Ea + Eb) - (Ea) The Bliss score is calculated using Eb), where E equals the effect (inhibition) of each individual agent (a and b). A difference greater than 10% between the observed percentage of inhibition and the predicted percentage of inhibition is considered synergistic. Bliss scores are reported only for combinations causing >20% growth inhibition (GDI value <80). Because Bliss is based on growth inhibition calculations, scores within the cytotoxic dose range may not accurately reflect synergistic cytotoxicity.

[0527] result

[0528] In 7 / 7 cell lines, as assessed by Bliss independence, synergistic growth inhibition was observed at multiple titration points in combinations of four structurally different SPLAMs with 1000 nM compound A (Tables 13A-13B, 14A-14E, 15A-15B, 16A-16E, 17A-17B, 18A-18E, and 19A-19B). In the context of each SPLAM, MTAP Defective cell lines showed greater growth inhibition and synergistic effects than wild-type cell lines at lower SPLAM doses. Only... MTAP Combined cytotoxicity was observed in the defective NCI-H838, SW900, and RT112 / 84 cell lines, but not in any wild-type cell lines.

[0529] Tables 13A-13B: Synergistic growth inhibition of two wild-type cell lines by the combination of indisulan and 1000 nM compound A

[0530] The synergistic effect of indisulan in the titration of 1000 nM compound A was measured by Bliss independence analysis. A Bliss score >10 was considered "...". The Bliss score indicates a synergistic effect. The Bliss score applies to combinations exhibiting a growth inhibition rate greater than 20% (GDI < 80). Synergistic effects reported using Bliss may be underrepresented in the cytotoxic dose range. Growth and death index values ​​for combination titration and each individual agent compound A (Cmpd A) and Indisulan are shown, ranging from -100 to 100, with the midpoint (0) representing the cell number at the start of the combination (T0). Scores less than 0 are highlighted in black and represent a cytotoxic response. Bliss and GDI values ​​for five MTAP-deficient cell lines are shown: NCI-H520 (13A), TCCSUP (13B).

[0531] Table 13A: NCI-H520

[0532] Table 13B: TCCSUP

[0533] Tables 14A-14E: Synergistic growth inhibition of five MTAP-deficient cell lines by the combination of E7820 and 1000 nM compound A

[0534] The synergistic effect of E7820 in the titration of 1000 nM compound A was measured by Bliss independence analysis. A Bliss score >10 was considered "..." The Bliss score indicates that the combination exhibits a growth inhibition rate greater than 20% (GDI < 80). Synergistic effects reported using Bliss may be underrepresented in the cytotoxic dose range. Growth and death index values ​​for combination titration and each individual agent compound A (Cmpd A) and Indisulan are shown, ranging from -100 to 100, with the midpoint (0) representing the cell count at the start of the combination (T0). A score less than 0 is indicated by the " “” indicates and represents cytotoxic response. Bliss and GDI values ​​for five MTAP-deficient cell lines are shown: NCI-H838 (14A), SW900 (14B), RT112 / 84 (14C), UMUC5 (14D), and UMUC11 (14E).

[0535] Table 14A: NCI-H838

[0536] Table 14B: SW900

[0537] Table 14C: RT112 / 84

[0538] Table 14D: UMUC5

[0539] Table 14E: UMUC11

[0540] Tables 15A-15B: Synergistic growth inhibition of two wild-type cell lines by the combination of E7820 and 1000 nM compound A.

[0541] The synergistic effect of E7820 in the titration of 1000 nM compound A was measured by Bliss independence analysis. A Bliss score >10 was considered "..." The Bliss score indicates that the combination exhibits a growth inhibition rate greater than 20% (GDI < 80). Synergistic effects reported using Bliss may be underrepresented in the cytotoxic dose range. Growth and death index values ​​for combination titration and each individual agent compound A (Cmpd A) and Indisulan are shown, ranging from -100 to 100, with the midpoint (0) representing the cell count at the start of the combination (T0). A score less than 0 is indicated by the " “” indicates and represents cytotoxic response. Bliss and GDI values ​​for five MTAP-deficient cell lines are shown: NCI-H520 (15A), TCCSUP (15B).

[0542] Table 15A: NCI-H520

[0543] Table 15B: TCCSUP

[0544] Tables 16A-16E: Synergistic growth inhibition of five MTAP-deficient cell lines by the combination of chloroquine sulfonamide and 1000 nM compound A

[0545] The synergistic effect of chloroquine sulfonamide in the titration of 1000 nM compound A was measured using Bliss independence analysis. A Bliss score >10 was considered "significant". The Bliss score indicates that the combination exhibits a growth inhibition rate greater than 20% (GDI < 80). Synergistic effects reported using Bliss may be underrepresented in the cytotoxic dose range. Growth and death index values ​​for combination titration and each individual agent compound A (Cmpd A) and Indisulan are shown, ranging from -100 to 100, with the midpoint (0) representing the cell count at the start of the combination (T0). A score less than 0 is indicated by the " “” indicates and represents the cytotoxic response. Bliss and GDI values ​​for five MTAP-deficient cell lines are shown: NCI-H838 (16A), SW900 (16B), RT112 / 84 (16C), UMUC5 (16D), and UMUC11 (16E).

[0546] Table 16A: NCI-H838

[0547] Table 16B: SW900

[0548] Table 16C: RT112 / 84

[0549] Table 16D: UMUC5

[0550] Table 16E: UMUC11

[0551] Tables 17A-17B: Synergistic growth inhibition of two wild-type cell lines by the combination of chloroquine sulfonamide and 1000 nM compound A.

[0552] The synergistic effect of chloroquine sulfonamide in the titration of 1000 nM compound A was measured using Bliss independence analysis. A Bliss score >10 was considered "significant". The Bliss score indicates that the combination exhibits a growth inhibition rate greater than 20% (GDI < 80). Synergistic effects reported using Bliss may be underrepresented in the cytotoxic dose range. Growth and death index values ​​for combination titration and each individual agent compound A (Cmpd A) and Indisulan are shown, ranging from -100 to 100, with the midpoint (0) representing the cell count at the start of the combination (T0). A score less than 0 is indicated by the " “” indicates and represents the cytotoxic response. Bliss and GDI values ​​for five MTAP-deficient cell lines are shown: NCI-H520 (17A), TCCSUP (17B).

[0553] Table 17A: NCI-H520

[0554] Table 17B: TCCSUP

[0555] Tables 18A-18E: Synergistic growth inhibition of five MTAP-deficient cell lines by the combination of tacisulan and 1000 nM compound A

[0556] The synergistic effect of Tasisulan in the titration of 1000 nM compound A was measured using Bliss independence analysis. A Bliss score >10 was considered "...". The Bliss score indicates that the combination exhibits a growth inhibition rate greater than 20% (GDI < 80). Synergistic effects reported using Bliss may be underrepresented in the cytotoxic dose range. Growth and death index values ​​for combination titration and each individual agent compound A (Cmpd A) and Indisulan are shown, ranging from -100 to 100, with the midpoint (0) representing the cell count at the start of the combination (T0). A score less than 0 is indicated by the " “” indicates and represents cytotoxic response. Bliss and GDI values ​​for five MTAP-deficient cell lines are shown: NCI-H838 (18A), SW900 (18B), RT112 / 84 (18C), UMUC5 (18D), and UMUC11 (18E).

[0557] Table 18A: NCI-H838

[0558] Table 18B: SW900

[0559] Table 18C: RT112 / 84

[0560] Table 18D: UMUC5

[0561] Table 18E: UMUC11

[0562] Tables 19A-19B: Synergistic growth inhibition of two wild-type cell lines by the combination of tacisulan and 1000 nM compound A.

[0563] The synergistic effect of Tasisulan in the titration of 1000 nM compound A was measured using Bliss independence analysis. A Bliss score >10 was considered "...". The Bliss score indicates that the combination exhibits a growth inhibition rate greater than 20% (GDI < 80). Synergistic effects reported using Bliss may be underrepresented in the cytotoxic dose range. Growth and death index values ​​for combination titration and each individual agent compound A (Cmpd A) and tacisulan are shown, ranging from -100 to 100, with the midpoint (0) representing the cell count at the start of the combination (T0). A score less than 0 is indicated by the " “” indicates and represents the cytotoxic response. Bliss and GDI values ​​for five MTAP-deficient cell lines are shown: NCI-H520 (19A), TCCSUP (19B).

[0564] Table 19A: NCI-H520

[0565] Table 19B: TCCSUP

[0566] Example 6: MAT2A and topoisomerase inhibitors in vitro MTAP Defect model provides combined benefits

[0567] Materials and methods

[0568] Use a set of 13 types MTAP This study evaluated the combined effects of a MAT2A inhibitor (compound A) and a topoisomerase inhibitor (10-hydroxycamptothecin, irinotecan, topotecan, daunorubicin, doxorubicin, and etoposide) in defective non-small cell lung cancer (NSCLC), bladder cancer, pancreatic cancer, gastric cancer, esophageal cancer, and head and neck squamous cell carcinoma (HNSCC) cancer cell lines. To determine the optimal concentration range for each compound in each cell line for the combined screening, cells were seeded at a density of 150 cells / well (compound A) or 500 cells / well (topoisomerase inhibitors) in 384-well plates. After 24 hours, cells were treated with a 9-point titration of each compound as a single agent and cultured at 37°C and 5% CO2 for 6 days. For the combined screening, cells were seeded at 150 cells / well in 384-well plates. After 24 hours, cells were co-treated with a 5-point titration of each compound in an optimized 6×6 dose matrix and cultured at 37°C and 5% CO2 for 6 days. Cells were then lysed using Promega CellTiter-Glo (CTG) 2.0 reagent according to the manufacturer's protocol, and chemiluminescence signals were measured using an EnVision plate reader (PerkinElmer) to quantify cell numbers. A plate of untreated cells was harvested at compound addition (T0 or time zero) to quantify cell numbers using CTG. Each data point was run three times in a technically reproducible manner.

[0569] The percentage of growth inhibition is calculated as follows: If T <V0:100 (1-(T-V0) / V0) If T≥V0:100 (1-(T-V0) / (V-V0)) Where T is the signal measurement of the test item, V is the control measurement of the mordant treatment, and V0 is the mordant control measurement at T0. The percentage of growth inhibition was used to generate dose-response curves and GI values ​​for single-drug activity and synergistic effects of drug combinations using Chalice Analyzer software (Horizon). 50 Calculations. Growth inhibition of 100% and >100% represents cell arrest and cytotoxicity, respectively.

[0570] Synergistic growth inhibition was assessed using the Loewe additive model. The observed growth inhibition at each dose was compared to the predicted inhibition based on the additive activity of any drug in combination with itself. The Loewe additive model was calculated as follows: (X / X) I ) +(Y / Y I I = 1 Loewe , where X I and Y I This refers to the single-drug effective concentration of the observed combined effect I. A difference of ≥20% between observed and predicted growth inhibition is considered synergistic.

[0571] The synergy score is calculated to quantify the strength of the synergy, as shown below: Synergy score = log f X log f Y Σ max(0,I 数据 (I) 数据 – I Loewe ) The inhibition score for each component agent and combination point in the matrix was calculated relative to the median of all untreated / codiluted control wells. A synergistic score > 2.22 was considered synergistic because it corresponded to high growth inhibition and exceeded the Loewe additive model by ≥ 20% at multiple dose points.

[0572] result

[0573] In a group of 13 MTAP In defective cell lines, combinations of compound A and six different topoisomerase inhibitors were tested. Synergistic growth inhibition was observed in each combination, but to varying degrees across the entire cell line group. For example, in 10 / 13 cells… MTAP In defective cell lines, the combination of compound A and 10-hydroxycamptothecin enhanced the observed growth inhibition compared to the single agent and showed a strong synergistic effect (determined by a synergistic score > 2.22). Figures 2A-1 to 2G , Figures 3A to 3G-2 (and Table 20).

[0574] In 8 / 13 MTAPIn defective cell lines, the combination of compound A and irinotecan also showed enhanced growth inhibition compared to the single agent, and exhibited a synergistic effect. Figures 4A-1 to 4G , Figures 5A to 5G-2 (and Table 20). In 6 / 13 MTAP In defective cell lines, the combination of compound A and topotecan also enhanced the growth inhibition observed with the single agent, showing a synergistic effect. Figures 6A-1 to 6G , Figures 7A to 7G-2 (and Table 20). In 12 / 13 MTAP In defective cell lines, the combination of compound A and daunorubicin showed enhanced growth inhibition and strong synergistic effects. Figures 8A-1 to 8G , Figures 9A to 9G-2 (and Table 20). In 8 / 13 MTAP In defective cell lines, the combination of compound A and doxorubicin showed enhanced growth inhibition and strong synergistic effects. Figures 10A-1 to 10G , Figures 11A to 11G-2 (and Table 20). In 5 / 13 MTAP In defective cell lines, the combination of compound A and etoposide showed enhanced growth inhibition and strong synergistic effects. Figures 12A-1 to 12G , Figures 13A to 13G-2 (and Table 20).

[0575] In the growth inhibition plot, growth inhibition is presented as the percentage of T0 for each cell line in a 6×6 dose matrix. 50-100% growth inhibition is indicated by… The figure indicates that 100% growth inhibition represents cell growth arrest. Growth inhibition greater than 100% is highlighted in dark gray and represents cytotoxicity. In the Loewe synergistic plot, the synergistic effect of compound A and 10-hydroxycamptothecin at all tested concentrations was measured using the Loewe additive model for each cell line. Values ​​>20 are indicated by the figure. This indicates that it has a synergistic effect.

[0576] Table 20: Combinations of Compound A with 10-hydroxycamptothecin (10-HCPT), irinotecan, topotecan, daunorubicin, doxorubicin, or etoposide in 13 different formulations. MTAP Synergistic effect score in defective cell lines.

[0577]

[0578] A synergistic score > 2.22 is considered to indicate synergistic effect.

[0579] Example 7: Antitumor activity of compound A and irinotecan in RT112 / 84 mouse xenograft model

[0580] All mouse studies were approved by the Institutional Animal Care and Use Committee (IACUC) in accordance with guidelines from the National Institutes of Health (NIH). Mice were housed under pathogen-free conditions and provided with food and water at will.

[0581] The antitumor effects of compound A and irinotecan hydrochloride as single agents or in combination were evaluated in a RT112 / 84 human bladder tumor cell-derived xenograft (CDX) model. Cells were expanded in RPMI-1640 (Gibco, catalog number 11875093) containing 10% fetal bovine serum. These cells were mycoplasma-free and validated by STR analysis. Five million cells in logarithmic growth phase were resuspended in Hanks balanced salt solution containing 50% Matrigel and subcutaneously implanted into the flank of each recipient female Cr1:NU-Foxn1nu mouse. Mice were housed in miniature isolation cages with corn cob bedding and additional enrichment consisting of sterile nesting material (Innovive) and Bio-huts (Bio-Serv). Water (Innovive) and diet (Teklad Global 19% Protein Extruded Diet 2919, irradiated) were provided at will. The environment is maintained at a 12-hour light cycle of approximately 68-72℉ and 40%-60% relative humidity.

[0582] Tumor volume (TV) is calculated using the following formula: TV (mm) 3 = (width × width × length) / 2. Tumor growth inhibition (TGI) is calculated as follows: [(TV control) 最终 TV therapy 最终 ) / (TV comparison) 最终 – TV Comparison 初始 [×100]. Statistical significance of TV was analyzed using GraphPadPrism version 10.0.3. Repeated measures two-way ANOVA and Tukey multiple comparisons were used, and p-values ​​were derived from data collected on day 14 (the end date of the mediator group), and were considered statistically significant if less than 0.05. Tumor regression was calculated as the percentage change in tumor volume of the final tumor volume measurement on day 73 compared to the initial tumor volume on day 1. Tumor regression was defined as a reduction in TV on day 73 compared to day 1. The tumor regression rate was defined as the percentage of tumor regression in each group of animals.

[0583] The average tumor volume at the start of drug administration was approximately 249 mm. 3Ten mice were randomly assigned to each treatment group. Mice were administered the mediator, 10 mg / kg of compound A orally once daily for 3 consecutive days followed by a 4-day rest period, and then intraperitoneally (IP) with 5 mg / kg of compound B, or a combination of compound A and compound B. The mediator group consisted of a combination of mediator A (for compound A, sterile water containing 0.5% 400 cps methylcellulose and 0.5% Tween-80) and mediator B (for irinotecan hydrochloride, saline).

[0584] result

[0585] Treatment with compound A alone resulted in a 53% TGI, while treatment with irinotecan hydrochloride alone resulted in an 83% TGI. The combination of compound A and irinotecan hydrochloride resulted in a 90% TGI, as shown in Table 21. Figure 14 As shown in Table 21, the combination of compound A and irinotecan hydrochloride increased the number of mice with tumor regression, producing a 50% response rate on day 73, compared to a 30% response rate for irinotecan hydrochloride alone. Figure 15 As shown.

[0586] Table 21: Overview of the antitumor activities of compounds A and B in RT112 / 84

[0587] Example 8: Antitumor activity of compound A and irinotecan in the MKN45 mouse xenograft model of gastric cancer

[0588] All mouse studies were approved by the Institutional Animal Care and Use Committee (IACUC) in accordance with guidelines from the National Institutes of Health (NIH). Mice were housed under pathogen-free conditions and provided with food and water at will.

[0589] The antitumor effects of compound A and irinotecan hydrochloride as single agents or in combination were evaluated in a MKN45 (RCB1001) human gastric tumor cell-derived xenograft (CDX) model. MKN45 cells were expanded in RPMI1640 containing 10% fetal bovine serum. Cells were mycoplasma-free and validated by STR analysis. Ten million cells in logarithmic growth phase were resuspended in Hanks balanced salt solution containing 50% Matrigel and subcutaneously implanted into the flank of each recipient female Cr1:NU-Foxn1nu mouse. Mice were housed in miniature isolation cages with corn cob bedding and additional enrichment consisting of sterile nesting material (Innovive) and Bio-huts (Bio-Serv). Water (Innovive) and diet (Teklad Global 19% Protein Extruded Diet 2919, irradiated) were provided at will. The environment is maintained at a 12-hour light cycle of approximately 68-72℉ and 40%-60% relative humidity.

[0590] Tumor volume (TV) is calculated using the following formula: TV (mm) 3 = (width × width × length) / 2. Tumor growth inhibition (TGI) is calculated as follows: [(TV control) 最终 TV therapy 最终 ) / (TV comparison) 最终 – TV Comparison 初始 [(×100)] The statistical significance of TV was analyzed using GraphPadPrism version 10.2.2. Repeated measures two-way ANOVA and Dunnet multiple comparisons were used, and p-values ​​were derived from data collected at the mediator group termination date, and a p-value less than 0.05 was considered statistically significant.

[0591] The average tumor volume at the start of drug administration was approximately 190 mm. 3 Six mice were randomly assigned to each treatment group. Mice were administered the mediator, compound A at 10 mg / kg, orally once daily for 3 consecutive days followed by a 4-day rest period, and then intraperitoneally (IP) with compound B at 5 mg / kg or 10 mg / kg, or a combination of compound A and compound B. The mediator group consisted of a combination of mediator A (for compound A, sterile water containing 0.5% 400 cps methylcellulose and 0.5% Tween-80) and mediator B (for irinotecan hydrochloride, saline).

[0592] result

[0593] Compound A alone resulted in a 56% TGI, while irinotecan hydrochloride alone resulted in a TGI ranging from 49% to 85%. The combination of compound A and irinotecan hydrochloride resulted in a 103% TGI, as shown in Table 22. Figure 16 As shown in Table 22, the combination of compound A and irinotecan hydrochloride increased the number of mice with tumor regression, producing an 83% tumor regression rate by day 25, compared to 0% to 17% for irinotecan hydrochloride alone.

[0594] Table 22: Overview of the antitumor activities of compounds A and B in MKN45

[0595] Example 9: Antitumor activity of compound A and irinotecan in LMSU mouse xenograft model of gastric cancer

[0596] All mouse studies were approved by the Institutional Animal Care and Use Committee (IACUC) in accordance with guidelines from the National Institutes of Health (NIH). Mice were housed under pathogen-free conditions and provided with food and water at will.

[0597] The antitumor effects of compound A and irinotecan hydrochloride as single agents or in combination were evaluated in an LMSU (RCB1062) human gastric tumor cell-derived xenograft (CDX) model. LMSU was expanded in HamF10 cells containing 10% fetal bovine serum. Cells were mycoplasma-free and validated by STR analysis. Ten million cells in logarithmic growth phase were resuspended in Hanks balanced salt solution containing 50% Matrigel and subcutaneously implanted into the flank of each recipient female Cr1:NU-Foxn1nu mouse. Mice were housed in miniature isolation cages with corn cob bedding and additional enrichment consisting of sterile nesting material (Innovive) and Bio-huts (Bio-Serv). Water (Innovive) and diet (Teklad Global 19% Protein Extruded Diet 2919, irradiated) were provided at will. The environment is maintained at a 12-hour light cycle of approximately 68-72℉ and 40%-60% relative humidity.

[0598] Tumor volume (TV) is calculated using the following formula: TV (mm) 3 = (width × width × length) / 2. Tumor growth inhibition (TGI) is calculated as follows: [(TV control) 最终 TV therapy 最终 ) / (TV comparison) 最终 – TV Comparison 初始[(×100)] The statistical significance of TV was analyzed using GraphPadPrism version 10.2.2. Repeated measures two-way ANOVA and Dunnet multiple comparisons were used, and p-values ​​were derived from data collected at the mediator group termination date, and a p-value less than 0.05 was considered statistically significant.

[0599] The average tumor volume at the start of drug administration was approximately 185 mm. 3 Seven mice were randomly assigned to each treatment group. Mice were administered the mediator, 10 mg / kg of compound A orally once daily for 3 consecutive days followed by a 4-day rest period, and then intraperitoneally (IP) with 5 mg / kg or 10 mg / kg of compound B, or a combination of compound A and compound B. The mediator group consisted of a combination of mediator A (for compound A, sterile water containing 0.5% 400 cps methylcellulose and 0.5% Tween-80) and mediator B (for irinotecan hydrochloride, saline).

[0600] result

[0601] Compound A alone did not result in a significant TGI, while irinotecan hydrochloride alone resulted in a TGI of 42% to 80%. The combination of compound A and irinotecan hydrochloride resulted in a 97% TGI, as shown in Table 23. Figure 17 As shown in Table 23, the combination of compound A and irinotecan hydrochloride increased the number of mice with tumor regression, producing a 29% tumor regression rate on day 25, compared to 0% to 14% tumor regression rate produced by irinotecan hydrochloride alone.

[0602] Table 23: Overview of the antitumor activities of compounds A and B in LMSU

[0603] Specific embodiments of the invention are described herein, including the best mode of carrying out the invention known to the inventors. Variations of the disclosed embodiments will be apparent to those skilled in the art upon reading the foregoing description, and it is expected that such variations will be appropriately employed by those skilled in the art. Therefore, the invention is intended to be practiced in a manner different from that specifically described herein, and the invention includes all modifications and equivalents of the subject matter set forth in the appended claims as permitted by applicable law. Furthermore, unless otherwise indicated herein or clearly contradicted by the context, the invention covers any combination of the foregoing elements in all its possible variations.

[0604] All patent applications, patents, and print publications cited herein are incorporated herein in their entirety, except for any definitions, subject matter disclaimers, or denials, and unless the incorporated material is inconsistent with the express disclosure herein, in which case the language of this disclosure shall prevail.

[0605] Other embodiments are described in the following claims.

Claims

1. A method of treating cancer in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and administering to the subject a therapeutically effective amount of a topoisomerase inhibitor, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I)； in X is either N or CH; R 3 C 1-6 Halogenated alkyl, halogenated or C 3-6 cycloalkyl; R 2 For -NR 4 R 5 ; R 4 It is hydrogen or C 1-6 alkyl; R 5 For hydrogen, C 1-6 Alkyl or C 3-6 cycloalkyl; and R 1 It is a phenyl group substituted with 0-2 halogen groups.

2. The method according to claim 1, wherein the topoisomerase inhibitor is a type I topoisomerase inhibitor.

3. The method according to claim 2, wherein the type I topoisomerase inhibitor is selected from the group consisting of: 10-hydroxycamptothecin, irinotecan, hexylresorcinol, esaxatecan, delutec, belotecone, and topotecan, or pharmaceutically acceptable salts thereof.

4. The method of claim 3, wherein the type I topoisomerase inhibitor is irinotecan or a pharmaceutically acceptable salt thereof.

5. The method according to claim 1, wherein the topoisomerase inhibitor is a type II topoisomerase inhibitor.

6. The method of claim 5, wherein the type II topoisomerase inhibitor is selected from the group consisting of daunorubicin, doxorubicin, and etoposide, or pharmaceutically acceptable salts thereof.

7. The method according to any one of claims 1 to 6, wherein the MAT2A inhibitor is administered simultaneously or sequentially with the topoisomerase inhibitor.

8. A method of treating cancer in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and administering to the subject a therapeutically effective amount of a splicing inhibitor sulfonamide (SPLAM), wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I)； in X is either N or CH; R 3 C 1-6 Halogenated alkyl, halogenated or C 3-6 cycloalkyl; R 2 For -NR 4 R 5 ; R 4 It is hydrogen or C 1-6 alkyl; R 5 For hydrogen, C 1-6 Alkyl or C 3-6 cycloalkyl; and R 1 It is a phenyl group substituted with 0-2 halogen groups.

9. The method of claim 8, wherein the SPLAM is indisulan or a pharmaceutically acceptable salt thereof.

10. The method of claim 8, wherein the SPLAM is selected from the group consisting of E7820, chloroquine sulfonamide, and tacisulan, or a pharmaceutically acceptable salt thereof.

11. The method according to any one of claims 8 to 10, wherein the MAT2A inhibitor is administered simultaneously or sequentially with the SPLAM.

12. A method of treating cancer in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and administering to the subject a therapeutically effective amount of a KIF18 inhibitor, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: (I)； in X is either N or CH; R 3 C 1-6 Halogenated alkyl, halogenated or C 3-6 cycloalkyl; R 2 For -NR 4 R 5 ; R 4 It is hydrogen or C 1-6 alkyl; R 5 For hydrogen, C 1-6 Alkyl or C 3-6 cycloalkyl; and R 1 It is a phenyl group substituted with 0-2 halogen groups.

13. The method of claim 12, wherein the KIF18 inhibitor is a KIF18A inhibitor.

14. The method of claim 13, wherein the KIF18A inhibitor is sovexib or a pharmaceutically acceptable salt thereof.

15. The method of any one of claims 12 to 14, wherein the MAT2A inhibitor and the KIF18 inhibitor are administered simultaneously or sequentially.

16. The method according to any one of claims 1 to 15, wherein X is N.

17. The method according to any one of claims 1 to 15, wherein X is CH.

18. The method according to any one of claims 1 to 17, wherein R 4 It is hydrogen and R 5 It is hydrogen or C 1-3 alkyl.

19. The method according to any one of claims 1 to 18, wherein R 5 It can be hydrogen or methyl.

20. The method according to any one of claims 1 to 19, wherein R 1 It is a phenyl group that has been substituted with chlorine.

21. The method according to any one of claims 1 to 20, wherein R 3 C 1-3 Halogenated alkyl or halogenated.

22. The method according to any one of claims 1 to 21, wherein R 3 It is trifluoromethyl or chlorine.

23. The method according to any one of claims 1 to 21, wherein R 3 It is cyclopropyl.

24. The method according to any one of claims 1 to 23, wherein the MAT2A inhibitor is selected from the group consisting of compounds in Table 1 or pharmaceutically acceptable salts thereof.

25. The method according to any one of claims 1 to 24, wherein the MAT2A inhibitor is compound A: Compound A Or its pharmaceutically acceptable salt.

26. The method according to any one of claims 1 to 24, wherein the MAT2A inhibitor is compound A1: Compound A1 Or its pharmaceutically acceptable salt.

27. The method according to any one of claims 1 to 26, wherein the cancer is selected from the group consisting of: leukemia, glioma, lung cancer, esophageal cancer, MTAP-deficient pancreatic ductal adenocarcinoma (PDAC), melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, anal cancer, gastric cancer, colon cancer, colorectal cancer, soft tissue sarcoma, non-Hodgkin lymphoma, gastric cancer, esophageal gastric cancer, malignant peripheral nerve sheath tumor, mesothelioma, salivary gland tumor, urothelial carcinoma, gastrointestinal cancer, and sarcoma.

28. The method according to any one of claims 1 to 27, wherein the cancer is a solid tumor or a hematologic cancer.

29. The method according to any one of claims 1 to 28, wherein the cancer is a solid malignant tumor.

30. The method according to any one of claims 1 to 29, wherein the cancer is characterized by reduced or absent MTAP gene expression, absence of the MTAP gene, reduced function of the MTAP protein, reduced or absent level of the MTAP protein, MTA accumulation, or a combination thereof.

31. Use of a MAT2A inhibitor in the manufacture of a medicament for treating cancer, wherein the MAT2A inhibitor is used in combination with a topoisomerase inhibitor.

32. Use of a MAT2A inhibitor in the manufacture of a medicament for treating cancer, wherein the MAT2A inhibitor is used in combination with SPLAM.

33. Use of a MAT2A inhibitor in the manufacture of a medicament for treating cancer, wherein the MAT2A inhibitor is used in combination with a KIF18 inhibitor.

34. The use as claimed in any one of claims 31 to 33, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

35. The use as described in claim 34, wherein the MAT2A inhibitor is compound A or a pharmaceutically acceptable salt thereof, or compound A1 or a pharmaceutically acceptable salt thereof.

36. A method for inhibiting tumor growth or slowing tumor growth rate in a subject with MTAP-deficient cancer, the method comprising administering a therapeutically effective amount of a MAT2A inhibitor and a therapeutically effective amount of a topoisomerase inhibitor, SPLAM, or KIF18 inhibitor.

37. The method of claim 36, wherein tumor growth is measured by changes in tumor volume from a first time point to a second time point.

38. The method of claim 37, wherein the tumor volume at the second time point has not increased compared to the first time point.

39. The method of claim 38, wherein the tumor volume decreases from the first time point to the second time point.

40. A method of treating cancer in a subject of need, the method comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and a therapeutically effective amount of a topoisomerase inhibitor, SPLAM, or a KIF18 inhibitor. The subjects in question had not previously received MAT2A inhibitor treatment.

41. The method of claim 40, wherein the treatment reduces the tumor growth rate compared to treatment with the MAT2A inhibitor alone for a similar time period.

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