Combination therapy including MAT2A inhibitors and topoisomerase inhibitors, splicing inhibitor sulfonamides, or KIF18 inhibitors.

Abstract

Provided herein are combinations of methionine adenosyltransferase II alpha (MAT2A) inhibitors with topoisomerase inhibitors, splicing inhibitor sulfonamides, or KIF18 inhibitors. Methods of using such combinations to treat diseases or disorders, such as cancer, are also provided.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims the interests of U.S. Provisional Application No. 63 / 495,911, filed on 13 April 2023, and U.S. Provisional Application No. 63 / 598,799, filed on 14 November 2023, each of which is incorporated herein by reference in whole for all purposes. [Background technology]

[0002] Cancer is a leading cause of death worldwide. The limitations of current therapeutic approaches, such as chemotherapy and immunotherapy, are that their cytotoxic effects are not limited to cancer cells, but can also cause harmful side effects in normal tissues.

[0003] Methionine adenosyltransferase 2A (MAT2A) is an enzyme that uses methionine (Met) and adenosine triphosphate (ATP) to produce s-adenosylmethionine (SAM). SAM is the primary intracellular methyl donor used to methylate several substrates, including DNA, RNA, and proteins. One methylase that utilizes SAM as a methyl donor is protein arginine N-methyltransferase 5 (PRMT5). PRMT5 activity requires SAM, but PRMT5 is competitively inhibited by 5'-methylthioadenosine (MTA). Since MTA is part of the methionine salvage pathway, intracellular MTA levels are kept low in processes initiated by methylthioadenosine phosphorylase (MTAP).

[0004] MTAP is located at a locus on chromosome 9 that is often deleted in cells from cancer patients originating from several tissues, including the central nervous system, pancreas, esophagus, bladder, and lungs (cBioPortal database). Loss of MTAP leads to MTA accumulation, and MTAP-deficient cells are more dependent on SAM production, and consequently on MAT2A activity, compared to cells expressing MTAP. In a screening of shRNA cell lines across approximately 400 cancer cell lines, knockdown of MAT2A in MTAP-deficient cells resulted in a higher rate of survival reduction compared to MTAP WT cells (see McDonald et.al. 2017 Cell 170, 577-592). Furthermore, inducible knockdown of the MAT2A protein reduced tumor growth in vivo (see Marjon et.al., 2016 Cell Reports 15(3), 574-587). These results suggest that MAT2A inhibitors may offer a new treatment option for cancer patients, including those with MTAP-deficient tumors.

[0005] DNA topoisomerases are enzymes found in the cell nucleus that regulate the topological state of DNA by catalyzing the cleavage and rejoining of DNA strands. Recent studies suggest that topoisomerases are also involved in regulating template supercoils during RNA transcription. There are two main classes of mammalian topoisomerases. DNA topoisomerase I catalyzes changes in the topological state of double-stranded DNA by performing transient single-strand cleavage-rejoining cycles. In contrast, mammalian topoisomerase II alters the DNA topology by inducing transient enzymatic cross-linking double-strand breaks, followed by traversal and resealing. The antitumor activity associated with topoisomerase-toxic drugs is related to their ability to stabilize the enzymatic DNA-cleavable complex. This drug-inducible stabilization of the enzymatic DNA-cleavable complex effectively converts the enzyme into a cytotoxic form.

[0006] Indislam is an aryl sulfonamide drug with selective anticancer activity. Its mechanism of action and the basis of its selectivity are unknown. Many cancer cell lines derived from the hematopoietic and lymphoid systems are sensitive to indislam, and this sensitivity correlates with DCAF15 expression levels. Two other sulfonamides clinically tested, tasislam and CQS, have the same mechanism of action as indislam. This class of drugs is commonly referred to as SPLAM (splicing inhibitor sulfonamides).

[0007] Kinesin is a molecular motor that plays a crucial role in cell division and the transport of intracellular vesicles and organelles. Mitotic kinesin plays a role in spindle formation, chromosome segregation, centrosome segregation, and several aspects of dynamics (as outlined in O. Rath and F. Kozielski, Nature Review Cancer, 12:527-39, 2012). Human kinesins are classified into 14 subfamilies based on sequence homology within the “motor domain.” The ATPase activity of this domain drives unidirectional movement along microtubules (MTs). The non-motor domains of these proteins are responsible for cargo attachment. The “cargo” can include any one of several different membranous organelles, signaling scaffolds, and chromosomes. Kinesin uses the energy from ATP hydrolysis to move cargo along polar microtubules. Therefore, kinesin is often referred to as a “plus-end” or “minus-end” directional motor.

[0008] The KIF18A gene belongs to the kinesin-8 subfamily and is a plus-end oriented motor. KIF18A is thought to influence the dynamics at the plus end of kinetochore microtubules, thereby regulating correct chromosome arrangement and spindle tension. Depletion of human KIF18A leads to spindle elongation, increased chromosomal oscillations during metaphase, and activation of the mitotic spindle formation checkpoint in HeLa cervical cancer cells (MIMayr, et al., Current Biology 17, 488-98, 2007). KIF18A is overexpressed in various types of cancer, including but not limited to colorectal cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer, and ovarian cancer. Furthermore, deletion, knockdown, or inhibition of the KIF18A gene affects the mitotic spindle apparatus in cancer cell lines. In particular, inhibition of KIF18A has been shown to induce mitotic cell arrest, which is a known vulnerability that can promote mitotic cell death through apoptosis, mitotic collapse, or multipolar lethality or death after interphase mitotic slippage. Therefore, there is strong interest in finding inhibitors of the KIF18A protein.

[0009] Despite significant recent advances in cancer treatment, individuals suffering from the effects of cancer still need more effective and / or better treatments. [Overview of the project]

[0010] Provided herein are combination products comprising a methionine adenosyltransferase II alpha (MAT2A) inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor. These combination products are useful for the treatment of various cancers, including solid tumors. They are also useful for the treatment of various diseases or disorders treatable by inhibiting MAT2A. Furthermore, these combination products are useful for the treatment of MAT2A-deficient tumors.

[0011] In one embodiment, the herein provides a combination of a MAT2A inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor.

[0012] In one embodiment, provided herein are a pharmaceutical composition comprising a therapeutically effective amount of a methionine adenosyltransferase II alpha (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.

[0013] In one embodiment, the foregoing provides a method for treating cancer in a subject in need thereof, 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 cancer in the subject.

[0014] In one embodiment, the foregoing provides a method for treating cancer in a subject in need of such treatment, comprising administering to the subject a combination comprising a MAT2A inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor, together with at least a pharmaceutically acceptable carrier, thereby treating the cancer in the subject.

[0015] In yet another embodiment, the cancer is characterized by decreased or deleted methylthioadenosine phosphorylase (MTAP) gene expression, deletion of the MTAP gene, impaired function of the MTAP protein, decreased or deleted levels of the MTAP protein, accumulation of MTA, or a combination thereof.

[0016] In one embodiment, the foregoing provides a method for treating cancer in a subject in need thereof, 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 cancer in the subject.

[0017] In one embodiment, the foregoing provides a method for treating a disease or disorder treatable by inhibiting MAT2A in a subject requiring such treatment, comprising administering to the subject a combination comprising a methionine adenosyltransferase II alpha (MAT2A) inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor, thereby treating the disease or disorder of the subject. In one embodiment, the disease or disorder is cancer.

[0018] In one embodiment, the foregoing provides a method for treating a disease or disorder treatable by inhibiting MAT2A in a subject requiring such treatment, comprising administering to the subject a combination comprising a MAT2A inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor, together with at least a pharmaceutically acceptable carrier, thereby treating the disease or disorder of the subject. In one embodiment, the disease or disorder is cancer.

[0019] In one embodiment, the MAT2A inhibitor is a compound of formula I: [ka] or a pharmaceutically acceptable salt thereof, where the variable part of formula (I) is defined as follows:

[0020] In another embodiment, the MAT2A inhibitor is compound A having the following structural formula: [ka] or a pharmaceutically acceptable salt thereof.

[0021] In another embodiment, the MAT2A inhibitor is compound A1 having the following structural formula: [ka] or a pharmaceutically acceptable salt thereof.

[0022] MAT2A inhibitors for use in the combination therapies described herein are described in WO2020 / 123395 (PCT / US19 / 65260). The general and specific compounds described herein are incorporated herein by reference and can be used to treat the cancers described herein.

[0023] In one embodiment, the KIF18 inhibitor is compound B: [ka] or its pharmaceutically acceptable salt. The chemical name of compound B is N-(2-(4,4-difluoropiperidine-1-yl)-6-methylpyrimidine-4-yl)-4-((2-hydroxyethyl)sulfonamide)-2-(6-azaspiro[2.5]octan-6-yl)benzamide. Compound B is also known as "sovilnesib" or "AMG650". Compound B also includes its pharmaceutically acceptable salt.

[0024] In another embodiment, SPLAM is: [ka] or its pharmaceutically acceptable salts. The chemical name of indislam is N-(3-chloro-1H-indole-7-yl)benzene-1,4-disulfonamide. Indislam also includes its pharmaceutically acceptable salts.

[0025] In yet another embodiment, SPLAM is E7820: [ka] or its pharmaceutically acceptable salt. The chemical name of E7820 is 3-cyano-N-(3-cyano-4-methyl-1H-indole-7-yl)benzenesulfonamide. E7820 also includes its pharmaceutically acceptable salt.

[0026] In yet another embodiment, SPLAM is a chloroquinoxaline sulfonamide: [ka] or its pharmaceutically acceptable salt. The chemical name of chloroquinoxalinesulfonamide is 4-amino-N-(5-chloroquinoxalines-2-yl)benzenesulfonamide, also known as "CQS". Chloroquinoxalinesulfonamide also includes its pharmaceutically acceptable salts.

[0027] In another embodiment, SPLAM is Tasilam: [ka] or its pharmaceutically acceptable salt. The chemical name of tasislam is N-((5-bromothiophen-2-yl)sulfonyl)-2,4-dichlorobenzamide. Tasislam also includes its pharmaceutically acceptable salt.

[0028] In one embodiment, the topoisomerase inhibitor is a type I topoisomerase inhibitor.

[0029] In another embodiment, the type I topoisomerase inhibitor is 10-hydroxycamptothecin: [ka] 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]indolidino[1,2-b]quinoline-3,14(4H)-dione. 10-hydroxycamptothecin also includes its pharmaceutically acceptable salts.

[0030] In yet another embodiment, the type I topoisomerase inhibitor is irinotecan: [ka] or its pharmaceutically acceptable salt. The chemical name of irinotecan is (S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-9-yl[1,4'-bipiperidine]-1'-carboxylate. Irinotecan also includes its pharmaceutically acceptable salt.

[0031] In yet another embodiment, the type I topoisomerase inhibitor is topotecan: [ka] or its pharmaceutically acceptable salt. The chemical name of topotecan is (S)-10-((dimethylamino)methyl)-4-ethyl-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione. Topotecan also includes its pharmaceutically acceptable salt.

[0032] In one embodiment, the topoisomerase inhibitor is a type II topoisomerase inhibitor.

[0033] In another embodiment, the type II topoisomerase inhibitor is daunorubicin: [ka] 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-tetrahydrotetracene-5,12-dione. Daunorubicin also includes its pharmaceutically acceptable salts.

[0034] In yet another embodiment, the type II topoisomerase inhibitor is doxorubicin: [ka] 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-tetrahydrotetracene-5,12-dione. Doxorubicin also includes its pharmaceutically acceptable salts.

[0035] In yet another embodiment, the type II topoisomerase inhibitor is etoposide: [ka] 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-tetrahydrofloo[3',4':6,7]naphtho[2,3-d][1,3]dioxol-6(5aH)-one. Etoposide also includes its pharmaceutically acceptable salts. [Brief explanation of the drawing]

[0036] [Figure 1A-1]This study demonstrates the growth inhibition of compound B (sovylnesib) as a monotherapy in MTAP-deficient cell lines. [Figure 1A-2] This study demonstrates the growth inhibition of compound B (sovylnesib) as a monotherapy in MTAP-deficient cell lines. [Figure 1A-3] This study demonstrates the growth inhibition of compound B (sovylnesib) as a monotherapy in MTAP-deficient cell lines. [Figure 1A-4] This study demonstrates the growth inhibition of compound B (sovylnesib) as a monotherapy in MTAP-deficient cell lines. [Figure 1A-5] This study demonstrates the growth inhibition of compound B (sovylnesib) as a monotherapy in MTAP-deficient cell lines. [Figure 1B-1] This study demonstrates growth inhibition by compound B (sovilnesib) as a monotherapy in wild-type cell lines. [Figure 1B-2] This study demonstrates growth inhibition by compound B (sovilnesib) as a monotherapy in wild-type cell lines. [Figure 2A-1] This study demonstrates growth inhibition of compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 2A-2] This study demonstrates growth inhibition of compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 2B-1] This study demonstrates growth inhibition of compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 2B-2] This study demonstrates growth inhibition of compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 2C-1] This study demonstrates growth inhibition of compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 2C-2] This study demonstrates growth inhibition of compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 2D-1] This study demonstrates growth inhibition of compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 2D-2] This study demonstrates growth inhibition of compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 2E-1] This study demonstrates growth inhibition of compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 2E-2] This study demonstrates growth inhibition of compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 2F-1] This study demonstrates growth inhibition of compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 2F-2] This study demonstrates growth inhibition of compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 2G] This study demonstrates growth inhibition of compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 3A] This study demonstrates the Loewe synergy between compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 3B-1] This study demonstrates the Loewe synergy between compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 3B-2] This study demonstrates the Loewe synergy between compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 3C-1] This study demonstrates the Loewe synergy between compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 3C-2] This study demonstrates the Loewe synergy between compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 3D-1] This study demonstrates the Loewe synergy between compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 3D-2] This study demonstrates the Loewe synergy between compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 3E-1] This study demonstrates the Loewe synergy between compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 3E-2] This study demonstrates the Loewe synergy between compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 3F-1] This study demonstrates the Loewe synergy between compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 3F-2] This study demonstrates the Loewe synergy between compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 3G-1] This study demonstrates the Loewe synergy between compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 3G-2] This study demonstrates the Loewe synergy between compound A and 10-hydroxycamptothecin in MTAP-deficient cell lines. [Figure 4A-1] This study demonstrates growth inhibition of compound A and irinotecan in MTAP-deficient cell lines. [Figure 4A-2] This study demonstrates growth inhibition of compound A and irinotecan in MTAP-deficient cell lines. [Figure 4B-1] This study demonstrates growth inhibition of compound A and irinotecan in MTAP-deficient cell lines. [Figure 4B-2] This study demonstrates growth inhibition of compound A and irinotecan in MTAP-deficient cell lines. [Figure 4C-1] This study demonstrates growth inhibition of compound A and irinotecan in MTAP-deficient cell lines. [Figure 4C-2] This study demonstrates growth inhibition of compound A and irinotecan in MTAP-deficient cell lines. [Figure 4D-1] This study demonstrates growth inhibition of compound A and irinotecan in MTAP-deficient cell lines. [Figure 4D-2] This study demonstrates growth inhibition of compound A and irinotecan in MTAP-deficient cell lines. [Figure 4E-1] This study demonstrates growth inhibition of compound A and irinotecan in MTAP-deficient cell lines. [Figure 4E-2] This study demonstrates growth inhibition of compound A and irinotecan in MTAP-deficient cell lines. [Figure 4F-1] This study demonstrates growth inhibition of compound A and irinotecan in MTAP-deficient cell lines. [Figure 4F-2]This study demonstrates growth inhibition of compound A and irinotecan in MTAP-deficient cell lines. [Figure 4G] This study demonstrates growth inhibition of compound A and irinotecan in MTAP-deficient cell lines. [Figure 5A] This study demonstrates the Loewe synergy between compound A and irinotecan in MTAP-deficient cell lines. [Figure 5B-1] This study demonstrates the Loewe synergy between compound A and irinotecan in MTAP-deficient cell lines. [Figure 5B-2] This study demonstrates the Loewe synergy between compound A and irinotecan in MTAP-deficient cell lines. [Figure 5C-1] This study demonstrates the Loewe synergy between compound A and irinotecan in MTAP-deficient cell lines. [Figure 5C-2] This study demonstrates the Loewe synergy between compound A and irinotecan in MTAP-deficient cell lines. [Figure 5D-1] This study demonstrates the Loewe synergy between compound A and irinotecan in MTAP-deficient cell lines. [Figure 5D-2] This study demonstrates the Loewe synergy between compound A and irinotecan in MTAP-deficient cell lines. [Figure 5E-1] This study demonstrates the Loewe synergy between compound A and irinotecan in MTAP-deficient cell lines. [Figure 5E-2] This study demonstrates the Loewe synergy between compound A and irinotecan in MTAP-deficient cell lines. [Figure 5F-1] This study demonstrates the Loewe synergy between compound A and irinotecan in MTAP-deficient cell lines. [Figure 5F-2] This study demonstrates the Loewe synergy between compound A and irinotecan in MTAP-deficient cell lines. [Figure 5G-1] This study demonstrates the Loewe synergy between compound A and irinotecan in MTAP-deficient cell lines. [Figure 5G-2] This study demonstrates the Loewe synergy between compound A and irinotecan in MTAP-deficient cell lines. [Figure 6A-1] This study demonstrates the inhibition of compound A and topotecan growth in MTAP-deficient cell lines. [Figure 6A-2] This study demonstrates the inhibition of compound A and topotecan growth in MTAP-deficient cell lines. [Figure 6B-1] This study demonstrates the inhibition of compound A and topotecan growth in MTAP-deficient cell lines. [Figure 6B-2] This study demonstrates the inhibition of compound A and topotecan growth in MTAP-deficient cell lines. [Figure 6C-1] This study demonstrates the inhibition of compound A and topotecan growth in MTAP-deficient cell lines. [Figure 6C-2] This study demonstrates the inhibition of compound A and topotecan growth in MTAP-deficient cell lines. [Figure 6D-1] This study demonstrates the inhibition of compound A and topotecan growth in MTAP-deficient cell lines. [Figure 6D-2] This study demonstrates the inhibition of compound A and topotecan growth in MTAP-deficient cell lines. [Figure 6E-1] This study demonstrates the inhibition of compound A and topotecan growth in MTAP-deficient cell lines. [Figure 6E-2] This study demonstrates the inhibition of compound A and topotecan growth in MTAP-deficient cell lines. [Figure 6F-1] This study demonstrates the inhibition of compound A and topotecan growth in MTAP-deficient cell lines. [Figure 6F-2] This study demonstrates the inhibition of compound A and topotecan growth in MTAP-deficient cell lines. [Figure 6G] This study demonstrates the inhibition of compound A and topotecan growth in MTAP-deficient cell lines. [Figure 7A] This study demonstrates the Loewe synergy between compound A and topotecan in MTAP-deficient cell lines. [Figure 7B-1] This study demonstrates the Loewe synergy between compound A and topotecan in MTAP-deficient cell lines. [Figure 7B-2] This study demonstrates the Loewe synergy between compound A and topotecan in MTAP-deficient cell lines. [Figure 7C-1] This study demonstrates the Loewe synergy between compound A and topotecan in MTAP-deficient cell lines. [Figure 7C-2]This study demonstrates the Loewe synergy between compound A and topotecan in MTAP-deficient cell lines. [Figure 7D-1] This study demonstrates the Loewe synergy between compound A and topotecan in MTAP-deficient cell lines. [Figure 7D-2] This study demonstrates the Loewe synergy between compound A and topotecan in MTAP-deficient cell lines. [Figure 7E-1] This study demonstrates the Loewe synergy between compound A and topotecan in MTAP-deficient cell lines. [Figure 7E-2] This study demonstrates the Loewe synergy between compound A and topotecan in MTAP-deficient cell lines. [Figure 7F-1] This study demonstrates the Loewe synergy between compound A and topotecan in MTAP-deficient cell lines. [Figure 7F-2] This study demonstrates the Loewe synergy between compound A and topotecan in MTAP-deficient cell lines. [Figure 7G-1] This study demonstrates the Loewe synergy between compound A and topotecan in MTAP-deficient cell lines. [Figure 7G-2] This study demonstrates the Loewe synergy between compound A and topotecan in MTAP-deficient cell lines. [Figure 8A-1] This study demonstrates growth inhibition of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 8A-2] This study demonstrates growth inhibition of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 8B-1] This study demonstrates growth inhibition of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 8B-2] This study demonstrates growth inhibition of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 8C-1] This study demonstrates growth inhibition of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 8C-2] This study demonstrates growth inhibition of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 8D-1] This study demonstrates growth inhibition of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 8D-2]This study demonstrates growth inhibition of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 8E-1] This study demonstrates growth inhibition of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 8E-2] This study demonstrates growth inhibition of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 8F-1] This study demonstrates growth inhibition of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 8F-2] This study demonstrates growth inhibition of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 8G] This study demonstrates growth inhibition of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 9A] This study demonstrates the Loewe synergistic effect of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 9B-1] This study demonstrates the Loewe synergistic effect of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 9B-2] This study demonstrates the Loewe synergistic effect of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 9C-1] This study demonstrates the Loewe synergistic effect of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 9C-2] This study demonstrates the Loewe synergistic effect of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 9D-1] This study demonstrates the Loewe synergistic effect of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 9D-2] This study demonstrates the Loewe synergistic effect of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 9E-1] This study demonstrates the Loewe synergistic effect of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 9E-2] This study demonstrates the Loewe synergistic effect of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 9F-1] This study demonstrates the Loewe synergistic effect of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 9F-2] This study demonstrates the Loewe synergistic effect of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 9G-1] This study demonstrates the Loewe synergistic effect of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 9G-2] This study demonstrates the Loewe synergistic effect of compound A and daunorubicin in MTAP-deficient cell lines. [Figure 10A-1] This study demonstrates growth inhibition of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 10A-2] This study demonstrates growth inhibition of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 10B-1] This study demonstrates growth inhibition of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 10B-2] This study demonstrates growth inhibition of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 10C-1] This study demonstrates growth inhibition of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 10C-2] This study demonstrates growth inhibition of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 10D-1] This study demonstrates growth inhibition of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 10D-2] This study demonstrates growth inhibition of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 10E-1] This study demonstrates growth inhibition of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 10E-2] This study demonstrates growth inhibition of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 10F-1] This study demonstrates growth inhibition of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 10F-2] This study demonstrates growth inhibition of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 10G] This study demonstrates growth inhibition of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 11A] This study demonstrates the Loewe synergistic effect of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 11B-1] This study demonstrates the Loewe synergistic effect of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 11B-2] This study demonstrates the Loewe synergistic effect of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 11C-1] This study demonstrates the Loewe synergistic effect of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 11C-2] This study demonstrates the Loewe synergistic effect of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 11D-1] This study demonstrates the Loewe synergistic effect of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 11D-2] This study demonstrates the Loewe synergistic effect of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 11E-1] This study demonstrates the Loewe synergistic effect of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 11E-2] This study demonstrates the Loewe synergistic effect of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 11F-1] This study demonstrates the Loewe synergistic effect of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 11F-2] This study demonstrates the Loewe synergistic effect of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 11G-1] This study demonstrates the Loewe synergistic effect of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 11G-2] This study demonstrates the Loewe synergistic effect of compound A and doxorubicin in MTAP-deficient cell lines. [Figure 12A-1] This study demonstrates the inhibition of growth by compound A and etoposide in MTAP-deficient cell lines. [Figure 12A-2] This study demonstrates the inhibition of growth by compound A and etoposide in MTAP-deficient cell lines. [Figure 12B-1]This study demonstrates the inhibition of growth by compound A and etoposide in MTAP-deficient cell lines. [Figure 12B-2] This study demonstrates the inhibition of growth by compound A and etoposide in MTAP-deficient cell lines. [Figure 12C-1] This study demonstrates the inhibition of growth by compound A and etoposide in MTAP-deficient cell lines. [Figure 12C-2] This study demonstrates the inhibition of growth by compound A and etoposide in MTAP-deficient cell lines. [Figure 12D-1] This study demonstrates the inhibition of growth by compound A and etoposide in MTAP-deficient cell lines. [Figure 12D-2] This study demonstrates the inhibition of growth by compound A and etoposide in MTAP-deficient cell lines. [Figure 12E-1] This study demonstrates the inhibition of growth by compound A and etoposide in MTAP-deficient cell lines. [Figure 12E-2] This study demonstrates the inhibition of growth by compound A and etoposide in MTAP-deficient cell lines. [Figure 12F-1] This study demonstrates the inhibition of growth by compound A and etoposide in MTAP-deficient cell lines. [Figure 12F-2] This study demonstrates the inhibition of growth by compound A and etoposide in MTAP-deficient cell lines. [Figure 12G] This study demonstrates the inhibition of growth by compound A and etoposide in MTAP-deficient cell lines. [Figure 13A] This study demonstrates the Loewe synergy between compound A and etoposide in MTAP-deficient cell lines. [Figure 13B-1] This study demonstrates the Loewe synergy between compound A and etoposide in MTAP-deficient cell lines. [Figure 13B-2] This study demonstrates the Loewe synergy between compound A and etoposide in MTAP-deficient cell lines. [Figure 13C-1] This study demonstrates the Loewe synergy between compound A and etoposide in MTAP-deficient cell lines. [Figure 13C-2] This study demonstrates the Loewe synergy between compound A and etoposide in MTAP-deficient cell lines. [Figure 13D-1]This study demonstrates the Loewe synergy between compound A and etoposide in MTAP-deficient cell lines. [Figure 13D-2] This study demonstrates the Loewe synergy between compound A and etoposide in MTAP-deficient cell lines. [Figure 13E-1] This study demonstrates the Loewe synergy between compound A and etoposide in MTAP-deficient cell lines. [Figure 13E-2] This study demonstrates the Loewe synergy between compound A and etoposide in MTAP-deficient cell lines. [Figure 13F-1] This study demonstrates the Loewe synergy between compound A and etoposide in MTAP-deficient cell lines. [Figure 13F-2] This study demonstrates the Loewe synergy between compound A and etoposide in MTAP-deficient cell lines. [Figure 13G-1] This study demonstrates the Loewe synergy between compound A and etoposide in MTAP-deficient cell lines. [Figure 13G-2] This study demonstrates the Loewe synergy between compound A and etoposide in MTAP-deficient cell lines. [Figure 14] This study demonstrates the antitumor activity of compound A and irinotecan, individually or in combination, in the RT-112 / 84 bladder CDX model. [Figure 15] This shows the changes in individual tumor volume after administration of irinotecan alone or a combination of compound A and irinotecan in the RT-112 / 84 CDX model. [Figure 16] This study demonstrates the antitumor activity of compound A and irinotecan in the gastric cancer CDX model MKN45. [Figure 17] This study demonstrates the antitumor activity of compound A and irinotecan in the gastric cancer CDX model LMSU. [Modes for carrying out the invention]

[0037] Provided herein are combination therapies comprising a methionine adenosyltransferase II alpha (MAT2A) inhibitor or a pharmaceutically acceptable salt thereof, and a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor, or a pharmaceutically acceptable salt thereof. These combination therapies are useful for the treatment of various cancers. In another embodiment, the combination therapy is useful for the treatment of any one of the MAT2A-related diseases. In yet another embodiment, the combination therapy is useful for the treatment of any disease or disorder treatable by inhibiting MAT2A.

[0038] definition The definitions of various terms used herein are listed below. These definitions apply to terms used throughout this specification and the claims unless they are limited individually or as part of a larger group in specific cases.

[0039] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art. In general, the nomenclature and experimental procedures used herein in cell culture, molecular genetics, organic chemistry, and peptide chemistry are well known and commonly used in the art.

[0040] As used herein, the articles “a” and “an” refer to a grammatical object of one or more (i.e., at least one). For example, “an element” means one or more elements. Furthermore, the use of the term “including,” as well as other forms such as “include,” “includes,” and “included,” is not limited to these.

[0041] As used herein, the term “about” is to be understood by those skilled in the art and to some extent depending on the context in which it is used. As used herein, when referring to a measurable value such as a quantity or length of time, the term “about” means to include variations of ±20% or ±10%, e.g., ±5%, ±1%, and ±0.1%, from the specified value, such variations being appropriate for carrying out the disclosed method.

[0042] As used herein and in the claims, “comprising” may include embodiments that “consist of” and “consisting essentially of.” The terms “comprise(s),” “include(s),” “having,” “has,” “may,” and “contain(s),” and their variations, as used herein, are intended to be open-ended transitional phrases, terms, or statements that require the presence of a specified component / step and permit the presence of other components / steps.

[0043] Note that ratios, concentrations, quantities, and other numerical data may be expressed in range form as described herein. Such range forms are used for convenience and conciseness and should therefore be interpreted flexibly to include not only the numerical values ​​explicitly stated as limits of the range, but also all individual numerical values ​​or subranges contained within that range, as if each numerical value and subrange were explicitly stated. In addition, the phrase "approximately x to y" includes "approximately x to approximately y".

[0044] As used herein, the terms “combination,” “therapeutic combination,” “pharmaceutical combination,” or “combination product” refer to either a fixed combination of one dosage unit form, a non-fixed combination of different dosage forms, or a kit of parts for combination administration in which two or more therapeutic agents can be administered simultaneously or separately at time intervals.

[0045] The term “combination therapy” refers to the administration of two or more therapeutic agents to treat a therapeutic condition or disorder described herein. Such administrations include substantially simultaneous co-administration of therapeutic agents, such as a single formulation having a certain ratio of active ingredients or separate formulations of each active ingredient (e.g., capsules and / or intravenous formulations). In addition, such administrations also include the use of various therapeutic agents approximately simultaneously, sequentially, or separately at different times. Whether the active ingredients are administered as a single formulation or as separate formulations, the drugs are administered to the same patient as part of the same treatment cycle. In any case, the treatment regimen produces a beneficial effect, e.g., a synergistic effect, on the treatment of the conditions or disorders described herein.

[0046] As used herein, the terms “to treat” or “to treat” mean to inhibit a disease, for example, inhibiting a disease, condition, or disorder in an individual experiencing or exhibiting the pathology or symptoms of the disease, condition, or disorder (i.e., stopping the further manifestation of the pathology and / or symptoms), or to improve a disease, for example, improving a disease, condition, or disorder in an individual experiencing or exhibiting the pathology or symptoms of the disease, condition, or disorder (i.e., improving the pathology and / or symptoms), for example, reducing the severity of the disease.

[0047] As used herein, the terms “prevent” or “prevent” mean, if the disorder or disease has not occurred, that there will be no occurrence of such disorder or disease, or, if the disorder or disease has already occurred, that there will be no further occurrence of such disorder or disease. The ability of the combination therapy provided herein to prevent some or all of the symptoms associated with the disorder or disease will also be considered.

[0048] As used herein, the terms “patient,” “individual,” or “subject” refer to human or non-human mammals. Examples of non-human mammals include livestock and pets, such as sheep, cattle, pigs, dogs, cats, and marine mammals. Preferably, the patient, subject, or individual is human.

[0049] As used herein, the terms “effective dose,” “pharmaceutically effective dose,” and “therapeutably effective dose” refer to an amount of an agent that is non-toxic but sufficient to produce a desired biological outcome. This outcome may be the reduction or alleviation of signs, symptoms, or causes of a disease, or any other desirable change in the biological system. The appropriate therapeutic dose in any individual case can be determined by a person skilled in the art using conventional experimental methods.

[0050] As used herein, the term “pharmaceutically acceptable” means a substance that does not impair the biological activity or properties of a compound and is relatively non-toxic, such as a carrier or diluent, that is, can be administered to an individual without causing undesirable biological effects or adversely interacting with any of the components of a composition containing the substance.

[0051] As used herein, the term “pharmaceutically acceptable salt” refers to a derivative of a disclosed compound in which the parent compound has been modified by converting the present acidic or basic moiety to its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; and alkali or organic salts of acidic residues such as carboxylic acids. Examples of pharmaceutically acceptable salts described herein include conventional non-toxic salts of parent compounds formed from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts considered herein can be synthesized by conventional chemical methods from parent compounds containing a basic or acidic moiety. Generally, such salts can be prepared by reacting the free acid or free base form of these compounds with a stoichiometric amount of a suitable base or acid in water, an organic solvent, or a mixture thereof. Generally, non-aqueous solvents such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are used. The term “pharmaceutically acceptable salt” is not limited to monosalts or 1:1 salts. For example, "pharmaceutically acceptable salts" include bis-salts such as dihydrochloric acid. Lists of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., 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 whole.

[0052] As used herein, the terms “composition” or “pharmaceutical composition” refer to a mixture of at least one compound and a pharmaceutically acceptable carrier. A pharmaceutical composition facilitates the administration of the composition to a patient or subject. Multiple techniques for administering compounds exist in the art, including but not limited to intravenous, oral, aerosol, parenteral, ocular, pulmonary, and topical administration.

[0053] As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable substance, composition, or carrier, such as a liquid or solid filler, stabilizer, dispersant, suspending agent, diluent, excipient, thickener, solvent, or encapsulating material, which is involved in transporting or delivering a compound useful to a patient in such a way that it can perform its intended function. Typically, such a construct is transported or delivered from one organ or part of the body to another organ or part of the body. Each carrier must be “acceptable” in the sense that it is compatible with the other components of a formulation containing the compound disclosed herein and is not harmful to the patient. Some examples of substances that can function as pharmaceutically acceptable carriers include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; celluloses and their derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; 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 glycerin, 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; ethyl alcohol; phosphate buffer; and other non-toxic and suitable substances used in pharmaceutical formulations.

[0054] As used herein, “pharmaceutically acceptable carrier” also includes any coating agents, antimicrobial and antifungal agents, and absorption retarders that are compatible with the activity and suitability of the compounds disclosed herein and are physiologically acceptable to the patient. Auxiliary active compounds may also be incorporated into the composition. Other additional components that may be included in the pharmaceutical composition are known in the art and are described, for example, in Remington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, PA), which is incorporated herein by reference.

[0055] As used herein, the term “single formulation” refers to a single carrier or vehicle formulated to deliver both therapeutically effective amounts of a therapeutic agent to a patient. A single vehicle is designed to deliver therapeutically effective amounts of each agent together with any pharmaceutically acceptable carrier or excipient. In some embodiments, the vehicle is a tablet, capsule, pill, or patch. In other embodiments, the vehicle is a solution or suspension.

[0056] As used herein, "methionine adenosyltransferase II alpha inhibitor" or "MAT2A inhibitor" means an agent that modulates the activity of MAT2A, or an agent that inhibits the production of S-adenosylmethionine (SAM) by methionine adenosyltransferase 2A (MAT2A).

[0057] The combinations of agents described herein may exhibit synergistic effects. As used herein, the term “synergistic effect” or “synergistic action” refers to the action of two agents, such as a MAT2A inhibitor and a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor, in which a certain effect, such as delaying the symptomatic progression or symptoms of cancer, is greater than the simple additive effect of administering each agent alone. The synergistic effect can be calculated using preferred 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. Pathol Pharmacol. 114:313-326 (1926)), and the average effect equation (Chou, TC and Talalay, P., Adv. Enzyme Regul. 22:27-55 (1984)). Applying each of the equations mentioned above to experimental data and creating corresponding graphs can be helpful in evaluating the combined effects of drugs. The corresponding graphs associated with the equations mentioned above are the concentration-effect curve, the isobologram curve, and the combination exponential curve, respectively.

[0058] In one embodiment, the herein provides a combination therapy comprising a therapeutically effective amount of a MAT2A inhibitor; and a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor. A “therapeutably effective amount” of the combination of agents (i.e., a combination of a MAT2A inhibitor and a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor) is an amount sufficient to produce an observable improvement from baseline to the clinically observable signs and symptoms of the disorder treated by the combination.

[0059] "Alkyl" means a straight-chain saturated monovalent hydrocarbon radical having 1 to 6 carbon atoms (i.e., C1-6 means 1 to 6 carbons) or a branched-chain saturated monovalent hydrocarbon radical having 3 to 6 carbon atoms (i.e., C3-6 means 3 to 6 carbons). Alkyl may contain any number of carbons, e.g., C 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 3-6 , C 4-5 , C 4-6 and C 5-6 etc. Examples of alkyl groups include methyl, ethyl, propyl, 2-propyl, butyl, pentyl, etc. It will be recognized by those skilled in the art that the term "alkyl" may include "alkylene" groups.

[0060] "Amino" means -NH2.

[0061] "Cycloalkyl" means a monocyclic monovalent hydrocarbon radical having 3 to 6 carbon atoms (e.g., C 3-6 cycloalkyl), which may be saturated or contain one double bond. Cycloalkyl may contain any number of carbons such as C 3-6 , C 4-6 , and C 5-6 etc. A partially unsaturated cycloalkyl group has one or more double bonds in the ring, but the cycloalkyl group is not aromatic. Examples of saturated monocyclic cycloalkyl rings include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0062] "Halo" means fluoro, chloro, bromo, or iodo, preferably fluoro or chloro.

[0063] "Haloalkyl" refers to alkyl radicals as defined above, including those substituted with 1 to 5 halogen atoms such as fluorine or chlorine, or those substituted with different halogens, such as -CH2Cl, -CF3, -CHF2, -CH2CF3, -CF2CF3, -CF(CH3)2, etc. When the alkyl is substituted only with fluoro, it may be called a fluoroalkyl. For alkyl groups, haloalkyl groups can have any preferred number of carbon atoms, e.g., C 1-6 It may have.

[0064] Provided herein are combination products comprising a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor, or a pharmaceutically acceptable salt thereof. These combination products are also referred to herein as combination therapies. The combination products are useful for the treatment of various cancers, including solid tumors. In another embodiment, the combination products are useful for the treatment of any one of the MAT2A-related diseases. In another embodiment, the combination products are useful for the treatment of diseases or disorders treatable by inhibiting MAT2A. In another embodiment, the combination products are useful for the treatment of MAT2A-deficient tumors.

[0065] In one embodiment, the herein provides a combination of a MAT2A inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor. In several embodiments, the herein provides a combination of a MAT2A inhibitor and a topoisomerase inhibitor. In several embodiments, the herein provides a combination of a MAT2A inhibitor and SPLAM. In several embodiments, the herein provides a combination of a MAT2A inhibitor and a KIF18 inhibitor.

[0066] This disclosure provides MAT2A inhibitors. In one embodiment, the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: [ka] (In the formula, X is either CH or N, R 3 Hello, C 1-6 Haloalkyl or C 3-6 It is a cycloalkyl, R 2 -NR 4 R 5 And, R 4 is hydrogen or C 1-6 It is alkyl, R 5 is hydrogen, C 1-6 Alkyl or C 3-6 It is a cycloalkyl, R 1 (where phenyl is substituted with 0-2 halos).

[0067] In one embodiment, X in formula (I) and its sub-embodiments is CH. In one embodiment, X in formula (I) and its sub-embodiments is N.

[0068] In yet another embodiment, formula (I) and R in its sub-embodiments 3 is a halo or C 1-6 It is a haloalkyl. In one embodiment, formula (I) and R in its sub-embodiments 3 is a halo. In one embodiment, formula (I) and R in its sub-embodiments 3 C 1-6 It is a haloalkyl. In one embodiment, formula (I) and R in its sub-embodiments 3 C 3-6 It is cycloalkyl. In one embodiment, formula (I) and R in its sub-embodiments 3 R is chloro, fluoro, bromo, -CH2Cl, -CF3, -CHF2, -CH2CF3, -CF2CF3, or -CF(CH3)2. In one embodiment, R in formula (I) and its sub-embodiments 3 is chloro or -CF3. In one embodiment, formula (I) and R in its sub-embodiments 3is chloro. In one embodiment, formula (I) and R in its sub-embodiments 3 It is -CF3.

[0069] In yet another embodiment, formula (I) and R in its sub-embodiments 4 is H. In one embodiment, formula (I) and R in its sub-embodiments 4 C 1-3 It is alkyl. In one embodiment, formula (I) and R in its sub-embodiments 4 These are methyl, ethyl, propyl, or isopropyl.

[0070] In yet another embodiment, formula (I) and R in its sub-embodiments 5 is H. In one embodiment, formula (I) and R in its sub-embodiments 5 C 1-3 It is alkyl. In one embodiment, formula (I) and R in its sub-embodiments 5 is methyl, ethyl, propyl, or isopropyl. In one embodiment, formula (I) and R in its sub-embodiments 5 C 3-6 It is a cycloalkyl group.

[0071] In some embodiments of formula (I) and its sub-embodiments, R 4 and R 5 These are H, respectively. In some embodiments of formula (I) and its sub-embodiments, R 4 and R 5 One of them is H, and the other is C 1-3 It is alkyl. In some embodiments of formula (I) and its sub-embodiments, 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-embodiments, R 4 and R 5 Independently, C 1-3 It is alkyl.

[0072] In yet another embodiment, R in formula (I) and its sub - embodiments 2 is -NH2, -NHC 1-3 alkyl, or -N(C 1-3 alkyl)2. In one embodiment, R in formula (I) and its sub - embodiments 2 is NH2, -NHMe, or -N(Me)2. In one embodiment, R in formula (I) and its sub - embodiments 2 is NH2. In one embodiment, R in formula (I) and its sub - embodiments 2 is -NHMe.

[0073] In one embodiment, R in formula (I) and its sub - embodiments 1 is phenyl, and the phenyl is unsubstituted or substituted with 1 - 2 halos. In yet another embodiment, R in formula (I) and its sub - embodiments 1 is unsubstituted phenyl. In one embodiment, R in formula (I) and its sub - embodiments 1 is phenyl substituted with 1 halo. In one embodiment, R in formula (I) and its sub - embodiments 1 is phenyl substituted with 1 halo selected from fluoro and chloro. In one embodiment, R in formula (I) and its sub - embodiments 1 is phenyl substituted with chloro. In one embodiment, R in formula (I) and its sub - embodiments 1 is phenyl substituted with 2 halos.

[0074] In yet another embodiment, the MAT2A inhibitor is selected from the group consisting of the compounds in Table 1 or their pharmaceutically acceptable salts.

Table 1

[0075] In another embodiment, the MAT2A inhibitor is Compound A:[[ID=4*]]

Chemical formula

[0076] In another embodiment, the MAT2A inhibitor is compound A1 having the following structural formula: [ka] or a pharmaceutically acceptable salt thereof.

[0077] The preparation and activity of the MAT2A inhibitors provided herein are disclosed in PCT / US2019 / 065260 (WO2020 / 123395), the entirety of which is incorporated herein by reference.

[0078] In one embodiment, the combination product comprises a MAT2A inhibitor and a KIF18 inhibitor. In another embodiment, the KIF18 inhibitor is sovilnesib or a pharmaceutically acceptable salt thereof. The preparation and activity of sovilnesib are disclosed in PCT / US2019 / 038169 (WO2020 / 132648), which is incorporated herein by reference in its entirety.

[0079] In yet another embodiment, the combination product includes a MAT2A inhibitor and SPLAM.

[0080] In yet another embodiment, SPLAM is indislam or a pharmaceutically acceptable salt thereof. The preparation and activity of E7820 are disclosed in PCT / JP1994 / 001487 (WO1995 / 007276) and US5,767,283, the entire contents thereof are incorporated herein by reference.

[0081] In one embodiment, SPLAM is E7820 or a pharmaceutically acceptable salt thereof. The preparation and activity of E7820 are disclosed in PCT / JP2000 / 001071 (WO2000 / 050395) and US6,469,043, the entire contents of which are incorporated herein by reference.

[0082] In another embodiment, SPLAM is a chloroquinoxaline sulfonamide or a pharmaceutically acceptable salt thereof. The preparation and activity of chloroquinoxaline sulfonamides are disclosed in PCT / US2000 / 000191 (WO2000 / 040269), the entire contents of which are incorporated herein by reference.

[0083] In yet another embodiment, SPLAM is tasislam or a pharmaceutically acceptable salt thereof. In yet another embodiment, SPLAM is tasislam sodium. The preparation and activity of tasislam are disclosed in PCT / US2002 / 031568 (WO2003 / 035629), the entire contents of which are incorporated herein by reference.

[0084] In one embodiment, the combined product includes a MAT2A inhibitor and a topoisomerase inhibitor.

[0085] In another embodiment, the topoisomerase inhibitor is a type I topoisomerase inhibitor.

[0086] In yet another embodiment, the type I topoisomerase inhibitor is 10-hydroxycamptothecin. The preparation and activity of 10-hydroxycamptothecin are disclosed in PCT / US1990 / 005986 (WO1991 / 005556), the entirety of which is incorporated herein by reference.

[0087] In yet another embodiment, the type I topoisomerase inhibitor is irinotecan. The preparation and activity of irinotecan are disclosed in PCT / US1992 / 009864 (WO1993 / 09782), the entire contents of which are incorporated herein by reference.

[0088] In one embodiment, the type I topoisomerase inhibitor is topotecan. The preparation and activity of topotecan are disclosed in PCT / US1992 / 001029 (WO1992 / 014470), the entire content of which is incorporated herein by reference in its entirety.

[0089] In one embodiment, the type I topoisomerase inhibitor is hexylresorcinol, exatecan, deruxtecan, and velotecan, or a pharmaceutically acceptable salt thereof.

[0090] In another embodiment, the topoisomerase inhibitor is a type II topoisomerase inhibitor.

[0091] In yet another embodiment, the type II topoisomerase inhibitor is daunorubicin. The preparation and activity of daunorubicin are disclosed in US4,138,480, the entire content of which is incorporated herein by reference in its entirety.

[0092] In yet another embodiment, the type II topoisomerase inhibitor is doxorubicin. The preparation and activity of doxorubicin are disclosed in US4,138,480, the entire content of which is incorporated herein by reference in its entirety.

[0093] In one embodiment, the type II topoisomerase inhibitor is etoposide. The preparation and activity of etoposide are disclosed in PCT / GB1983 / 000257 (WO1984 / 001506), the entire content of which is incorporated herein by reference in its entirety.

[0094] Treatment method In one embodiment, provided herein is a method of treating cancer in a subject that needs it, 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:

Chemical formula

[0095] In another embodiment, provided herein is a method for treating cancer in a subject in need thereof, 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.

[0096] In one embodiment, the KIF18 inhibitor is a KIF18A inhibitor. In another embodiment, the KIF18A inhibitor is sovilnesib or a pharmaceutically acceptable salt thereof. In yet another embodiment, the MAT2A inhibitor is administered concurrently or sequentially with the KIF18 inhibitor.

[0097] In another embodiment, provided herein is a method for treating cancer in a subject in need thereof, 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.

[0098] In another embodiment, provided herein is a method for treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and 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.

[0099] In yet another embodiment, SPLAM is indislam or a pharmaceutically acceptable salt thereof. In yet another embodiment, SPLAM is selected from the group consisting of E7820, chloroquinoxaline sulfonamide, and tasislam, or a pharmaceutically acceptable salt thereof. In one embodiment, the MAT2A inhibitor is administered concurrently or sequentially with SPLAM.

[0100] In another embodiment, provided herein is a method for treating cancer in a subject in need thereof, 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.

[0101] In another embodiment, provided herein is a method for treating cancer in a subject in need thereof, 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.

[0102] In another embodiment, provided herein is a method for treating cancer in a subject in need thereof, comprising administering to the subject an antibody-drug conjugate (ADC) comprising a MAT2A inhibitor and a topoisomerase inhibitor, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0103] In yet another embodiment, the topoisomerase inhibitor is a type I topoisomerase inhibitor. In yet another embodiment, the type I topoisomerase inhibitor is selected from the group consisting of 10-hydroxycamptothecin, irinotecan, and topotecan, or a pharmaceutically acceptable salt thereof. In yet another embodiment, the type I topoisomerase inhibitor is selected from the group consisting of hexylresorcinol, exatecan, deruxtecan, and belotecan, or a pharmaceutically acceptable salt thereof.

[0104] In yet another embodiment, the ADC comprising a topoisomerase inhibitor is a type I topoisomerase inhibitor. In yet another embodiment, the ADC comprising a topoisomerase inhibitor is fam-trastuzumab deruxtecan-nxki. In yet another embodiment, the ADC comprising a topoisomerase inhibitor is AZD8205. In yet another embodiment, the ADC comprising a topoisomerase inhibitor is DS-1062 (also known as datopotamab deruxtecan).

[0105] 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 a pharmaceutically acceptable salt thereof. In yet another embodiment, the type II topoisomerase inhibitor is amsacrine (m-AMSA) or a pharmaceutically acceptable salt thereof. In yet another embodiment, the MAT2A inhibitor is administered simultaneously or sequentially with the topoisomerase inhibitor.

[0106] [[ID=]12]With respect to MAT2A of formula (I), in one embodiment, X is N. In another embodiment, X is CH.

[0107] With respect to MAT2A of formula (I), in yet another embodiment, R 4 is hydrogen and R 5 is hydrogen or C 1-3 alkyl. In yet another embodiment, R 5It is either hydrogen or methyl.

[0108] Regarding MAT2A in formula (I), in one embodiment, R 1 This is a phenyl compound substituted with chloroform.

[0109] With respect to MAT2A in formula (I), in another embodiment, R 3 C 1-3 In yet another embodiment, R 3 is trifluoromethyl or chloro. In yet another embodiment, R 3 It is cyclopropyl.

[0110] In one embodiment, the MAT2A inhibitor is selected from the group consisting of the 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.

[0111] In one embodiment, a method for treating cancer in a subject in need of treatment is provided herein, comprising administering to the subject a combination comprising a MAT2A inhibitor and a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor, together with at least a pharmaceutically acceptable carrier, thereby treating the cancer in the subject. In several embodiments, a method for treating cancer in a subject in need of treatment is provided herein, comprising administering to the subject a combination comprising a MAT2A inhibitor and a topoisomerase inhibitor, together with at least a pharmaceutically acceptable carrier, thereby treating the cancer in the subject. In several embodiments, a method for treating cancer in a subject in need of treatment is provided herein, comprising administering to the subject a combination comprising a MAT2A inhibitor and a splicing inhibitor sulfonamide (SPLAM), together with at least a pharmaceutically acceptable carrier, thereby treating the cancer in the subject. In some embodiments, the foregoing provides a method for treating cancer in a subject in need of such treatment, comprising administering to the subject a combination comprising a MAT2A inhibitor and a KIF18 inhibitor together with at least a pharmaceutically acceptable carrier, thereby treating the cancer in the subject.

[0112] In yet another embodiment, the cancer is characterized by decreased or deleted methylthioadenosine phosphorylase (MTAP) gene expression, deletion of the MTAP gene, impaired function of the MTAP protein, decreased or deleted levels of the MTAP protein, accumulation of MTA, or a combination thereof.

[0113] In one embodiment, a method for treating cancer in a subject in need is provided herein, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition containing a MAT2A inhibitor and a therapeutically effective amount of a pharmaceutical composition containing a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor, thereby treating the cancer in the subject. In several embodiments, a method for treating cancer in a subject in need is provided herein, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition containing a MAT2A inhibitor and a therapeutically effective amount of a pharmaceutical composition containing a topoisomerase inhibitor, thereby treating the cancer in the subject. In several embodiments, a method for treating cancer in a subject in need is provided herein, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition containing a MAT2A inhibitor and a therapeutically effective amount of a pharmaceutical composition containing a splicing inhibitor sulfonamide (SPLAM), thereby treating the cancer in the subject. In some embodiments, the methods provided herein for treating cancer in a subject in need include 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 cancer in the subject.

[0114] 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 for the manufacture of a pharmaceutical is provided. 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 sovilnesib. In another embodiment, SPLAM is indislam. In yet another embodiment, SPLAM is selected from the group consisting of E7820, chloroquinoxaline sulfonamide, and tasislam, 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.

[0115] In one embodiment of the method, the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: [ka] (In the formula, the variable part is defined above.)

[0116] In another embodiment of the method, the MAT2A inhibitor is selected from the group consisting of the compounds in Table 1 or pharmaceutically acceptable salts thereof.

[0117] In yet another embodiment of the method, the MAT2A inhibitor is compound A or a pharmaceutically acceptable salt thereof.

[0118] In yet another embodiment of the method, the MAT2A inhibitor is compound A1 or a pharmaceutically acceptable salt thereof.

[0119] In one embodiment of the method, 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.

[0120] 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 preparation for simultaneous, separate, or sequential use in a pharmaceutical. In one embodiment, a product is provided comprising a MAT2A inhibitor and a topoisomerase inhibitor as a combination preparation for simultaneous, separate, or sequential use in a pharmaceutical. In one embodiment, a product is provided comprising a MAT2A inhibitor and a splicing inhibitor sulfonamide (SPLAM) as a combination preparation for simultaneous, separate, or sequential use in a pharmaceutical. In another embodiment, a product is provided comprising a MAT2A inhibitor and a KIF18 inhibitor as a combination preparation for simultaneous, separate, or sequential use in a pharmaceutical. 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 sovilnesib. In another embodiment, SPLAM is indislam. In yet another embodiment, SPLAM is selected from the group consisting of E7820, chloroquinoxaline sulfonamide, and tasislam, 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 yet another embodiment, the topoisomerase inhibitor is selected from the group consisting of daunorubicin, doxorubicin, and etoposide, or pharmaceutically acceptable salts thereof.

[0121] In yet 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, soft tissue sarcoma, anal cancer, gastric cancer, colon cancer, colorectal cancer, soft tissue sarcoma, non-Hodgkin lymphoma, gastric cancer, esophageal gastric cancer, esophageal cancer, malignant peripheral nerve sheath tumor, and mesothelioma.

[0122] 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 colon or rectal adenocarcinoma. In one embodiment, the cancer is breast cancer. In one embodiment, the cancer is mammary adenocarcinoma. In one embodiment, the cancer is gastric 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.

[0123] In one embodiment, the cancer is characterized by being MTAP null.

[0124] In one embodiment, the cancer is characterized by MTAP deficiency.

[0125] In yet another embodiment, the cancer is a solid tumor. In yet another embodiment, the cancer is an MTAP-deficient solid tumor. In yet another embodiment, the cancer is a metastatic MTAP-deficient solid tumor.

[0126] In another embodiment, the cancer is metastatic.

[0127] In yet another embodiment, the cancer is a solid tumor.

[0128] In another embodiment, the cancer is MTAP-deficient lung cancer or MTAP-deficient pancreatic cancer, such as MTAP-deficient NSCLC, MTAP-deficient pancreatic ductal adenocarcinoma (PDAC), or MTAP-deficient esophageal cancer.

[0129] In another embodiment, the cancer is a tumor having an MTAP gene deletion.

[0130] In any one embodiment of this specification, cancer is a solid tumor or a hematological cancer. In one embodiment, the tumor is characterized by a deficiency of MTAP. In another embodiment, the tumor is characterized by normal expression of MTAP.

[0131] In yet 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 yet another embodiment, the cancer is NSCLC, esophageal and gastric cancer, and pancreatic cancer. In yet another embodiment, the cancer is bladder cancer or gastrointestinal cancer.

[0132] In yet another embodiment, the cancer is characterized by decreased or deleted MTAP gene expression, deletion of the MTAP gene, impaired function of the MTAP protein, decreased or deleted levels of the MTAP protein, accumulation of MTA, or a combination thereof.

[0133] In another embodiment, the cancer is characterized by a decrease or deletion of MTAP gene expression.

[0134] In another embodiment, cancer is characterized by impaired function of the MTAP protein.

[0135] In another embodiment, the cancer is characterized by a decrease or deletion of MTAP protein levels.

[0136] In another embodiment, cancer is characterized by the accumulation of MTA.

[0137] In yet another embodiment, the cancer is a tumor having PRKC fusion.

[0138] In one embodiment, the herein provides a method for inhibiting tumor growth or slowing the rate of tumor growth in a subject having MTAP-deficient cancer, the method comprising administering a therapeutically effective amount of a MAT2A inhibitor and a therapeutically effective amount of a topoisomerase inhibitor.

[0139] In another embodiment, provided herein is a method for inhibiting tumor growth or slowing the rate of tumor growth in a subject having MTAP-deficient cancer, comprising the administration of a therapeutically effective amount of a MAT2A inhibitor and a therapeutically effective amount of SPLAM.

[0140] In yet another embodiment, provided herein is a method for inhibiting tumor growth or slowing the rate of tumor growth in a subject having MTAP-deficient cancer, comprising the administration of a therapeutically effective amount of a MAT2A inhibitor and a therapeutically effective amount of a KIF18 inhibitor.

[0141] Tumor growth is generally measured by the change in tumor volume from a first time point to a second time point. In one embodiment, tumor growth is measured by the change from the tumor volume at the first time point to the tumor volume at the second time point. In some embodiments, the tumor volume at the second time point has not increased compared to the first time point. In some embodiments, the tumor volume has decreased from the first time point to the second time point.

[0142] In one embodiment, the herein provides a method for treating cancer in a subject in need thereof, 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 been previously treated with a MAT2A inhibitor. In some embodiments, the treatment reduces the rate of tumor growth compared to treatment with a MAT2A inhibitor alone for a similar period.

[0143] In another embodiment, provided herein is a method for treating cancer in a subject in need thereof, 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 been previously treated with a MAT2A inhibitor. In some embodiments, the treatment reduces the rate of tumor growth compared to treatment with a MAT2A inhibitor alone for a similar period.

[0144] In yet another embodiment, provided herein is a method for treating cancer in a subject in need thereof, 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 been previously treated with a MAT2A inhibitor. In some embodiments, the treatment reduces the rate of tumor growth compared to treatment with a MAT2A inhibitor alone for a similar period.

[0145] 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 one embodiment, the MAT2A inhibitor and the topoisomerase inhibitor are in separate dosage forms. In one embodiment, the MAT2A inhibitor and the splicing inhibitor sulfonamide (SPLAM) are in separate dosage forms. In another embodiment, the MAT2A inhibitor and the KIF18 inhibitor are in separate dosage forms.

[0146] In another embodiment, the MAT2A inhibitor and the topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or 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.

[0147] In another embodiment, the treatment comprises administering substantially simultaneously a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof. In yet another embodiment, the treatment comprises administering a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor or a pharmaceutically acceptable salt thereof at different times.

[0148] In yet another embodiment, a MAT2A inhibitor or a pharmaceutically acceptable salt thereof is administered to the subject, 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, a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor, or a pharmaceutically acceptable salt thereof is administered to the subject, followed by the administration of a MAT2A inhibitor or a pharmaceutically acceptable salt thereof.

[0149] 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.

[0150] 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 the administration of SPLAM. In some embodiments, the MAT2A inhibitor is administered after the administration of SPLAM.

[0151] 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.

[0152] In yet another embodiment, the method includes administering a MAT2A inhibitor to a subject requiring a MAT2A inhibitor.

[0153] In yet another embodiment, the method includes administering a topoisomerase inhibitor, a splicing inhibitor sulfonamide (SPLAM), or a KIF18 inhibitor to a subject requiring such an inhibitor.

[0154] In one embodiment, a MAT2A inhibitor or a pharmaceutically acceptable salt thereof and a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or 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 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.

[0155] In another embodiment, 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, soft tissue sarcoma, anal cancer, gastric cancer, colon cancer, colorectal cancer, soft tissue sarcoma, non-Hodgkin lymphoma, gastric cancer, esophageal gastric cancer, esophageal cancer, malignant peripheral nerve sheath tumor, and mesothelioma.

[0156] In one embodiment, the foregoing provides MAT2A inhibitors or pharmaceutically acceptable salts thereof, and topoisomerase inhibitors, splicing inhibitor sulfonamides (SPLAMs), or KIF18 inhibitors or pharmaceutically acceptable salts thereof for therapeutic use.

[0157] In one embodiment, MAT2A inhibitors or pharmaceutically acceptable salts thereof, and topoisomerase inhibitors, splicing inhibitor sulfonamides (SPLAMs), or KIF18 inhibitors, or pharmaceutically acceptable salts thereof, are intended for use in treating cancer in subjects requiring such treatment.

[0158] Exemplary lengths of time associated with a "cool" of the treatment methods disclosed herein include about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, about 12 weeks, about 13 weeks, about 14 weeks, about 15 weeks, about 16 weeks, about 17 weeks, about 18 weeks, about 19 weeks, about 20 weeks, about 21 weeks, about 22 weeks, about 23 weeks, about 24 weeks, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months, about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 19 months, about 20 months, about 21 months, about 22 months, about 23 months, about 24 months, about 30 months, about 3 years, about 4 years, and about 5 years.

[0159] In one embodiment of the method, the method involves administering a combination or composition comprising a therapeutically effective amount of the compound provided herein or a pharmaceutically acceptable salt thereof to a subject in need of treatment (including, but not limited to, humans or animals) (including subjects identified as needing treatment).

[0160] In another embodiment of the method, the treatment comprises co-administration of the MAT2A inhibitor or a pharmaceutically acceptable salt thereof with the topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor, or a pharmaceutically acceptable salt thereof. In one embodiment, the MAT2A inhibitor or a pharmaceutically acceptable salt thereof and the 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 yet another embodiment, the MAT2A inhibitor or a pharmaceutically acceptable salt thereof and the topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor, or a pharmaceutically acceptable salt thereof are in separate formulations or unit dosage forms.

[0161] In the aforementioned method, treatment may include administering substantially simultaneously the amount of the MAT2A inhibitor or a pharmaceutically acceptable salt thereof and the amount of the topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor or a pharmaceutically acceptable salt thereof, or administering the amount of the MAT2A inhibitor or a pharmaceutically acceptable salt thereof and the amount of the topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor or a pharmaceutically acceptable salt thereof at different times. In some embodiments of the aforementioned method, the amount of the MAT2A inhibitor or a pharmaceutically acceptable salt thereof and / or the amount of the topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor or a pharmaceutically acceptable salt thereof is administered in a dose that is ineffective when one or both of the MAT2A inhibitor or a pharmaceutically acceptable salt thereof and the topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor or a pharmaceutically acceptable salt thereof are administered alone, but is effective when administered in combination.

[0162] Pharmaceutical composition In one embodiment, the herein 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.

[0163] 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 is provided comprising a therapeutically effective amount of a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor or a pharmaceutically acceptable salt thereof.

[0164] In another embodiment, provided herein is 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, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor or a pharmaceutically acceptable salt thereof.

[0165] 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.

[0166] In one embodiment, the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: [ka] (In the formula, the variable part is defined above.)

[0167] In another embodiment, the MAT2A inhibitor is selected from the group consisting of the compounds in Table 1 or pharmaceutically acceptable salts thereof.

[0168] In another embodiment, the MAT2A inhibitor is compound A: [ka] or a pharmaceutically acceptable salt thereof.

[0169] In another embodiment, the MAT2A inhibitor is compound A1: [ka] or a pharmaceutically acceptable salt thereof.

[0170] In one embodiment, the KIF18 inhibitor is sovilnesib. In another embodiment, SPLAM is indislam. In yet another embodiment, SPLAM is selected from the group consisting of E7820, chloroquinoxaline sulfonamide, and tasislam, 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 yet another embodiment, the topoisomerase inhibitor is selected from the group consisting of daunorubicin, doxorubicin, and etoposide, or pharmaceutically acceptable salts thereof.

[0171] In yet another embodiment, provided herein is 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.

[0172] In one embodiment, the pharmaceutical composition is for use in treating cancer in a patient. In one embodiment, the pharmaceutical composition is for use in treating solid tumors in a patient. In another embodiment, the pharmaceutical composition is for use in treating solid malignant tumors in a patient. In yet another embodiment, the cancer is MTAP-deficient lung cancer or MTAP-deficient pancreatic cancer, for example, MTAP-deficient NSCLC or MTAP-deficient PDAC or MTAP-deficient esophageal cancer.

[0173] In any one embodiment of this specification, cancer is a solid tumor or a hematological cancer. In yet another embodiment, cancer is NSCLC, mesothelioma, squamous cell carcinoma of the head and neck, salivary gland tumor, urothelial carcinoma, sarcoma, or ovarian cancer. In yet another embodiment, cancer is NSCLC, esophageal and gastric cancer, and pancreatic cancer. In yet another embodiment, cancer is bladder cancer or gastrointestinal cancer. In yet another embodiment, cancer is bladder (urothelial) cancer or gastrointestinal cancer. In one embodiment, cancer is bladder cancer. In another embodiment, cancer is urothelial carcinoma. In yet another embodiment, cancer is gastrointestinal cancer.

[0174] In one embodiment, the pharmaceutical composition is for use in the treatment of mesothelioma in a patient. In one embodiment, the pharmaceutical composition is for use in the treatment of non-small cell lung cancer in a patient. In one embodiment, the pharmaceutical composition is for use in the treatment of non-squamous non-small cell lung cancer in a patient. In one embodiment, the pharmaceutical composition is for use in the treatment of colon cancer in a patient. In one embodiment, the pharmaceutical composition is for use in the treatment of rectal cancer in a patient. In one embodiment, the pharmaceutical composition is for use in the treatment of colonic adenocarcinoma or rectal adenocarcinoma of the colon or rectum in a patient. In one embodiment, the pharmaceutical composition is for use in the treatment of breast cancer in a patient. In one embodiment, the pharmaceutical composition is for use in the treatment of mammary gland cancer in a patient. In one embodiment, the pharmaceutical composition is for use in the treatment of gastric cancer in a patient. In one embodiment, the pharmaceutical composition is for use in the treatment of gastric adenocarcinoma in a patient. In one embodiment, the pharmaceutical composition is for use in the treatment of pancreatic cancer in a patient. In one embodiment, the pharmaceutical composition is for use in the treatment of pancreatic adenocarcinoma in a patient. In one embodiment, the pharmaceutical composition is for use in the treatment of bladder cancer in a patient.

[0175] Dosage / Prescription Combination administration includes administering the combination as a single formulation or unit dosage form, administering the individual agents of the combination simultaneously but separately, or administering the individual agents of the combination sequentially via any preferred route. The dosage of the individual agents of the combination may require that one agent(s) be administered more frequently than the other(s) of the combination. Therefore, to enable appropriate administration, a packaged pharmaceutical product may contain one or more dosage forms containing the combination of agents, and one or more dosage forms containing one of the agents of the combination but not the other(s) of the combination.

[0176] The actual dose level of the active ingredient in the pharmaceutical composition may be varied to obtain an amount of the active ingredient that is effective in achieving the desired therapeutic response for a particular patient, composition, and mode of administration without causing toxicity to the patient.

[0177] In particular, the level of dosage selected depends on a variety of factors, including the activity of the specific compound being employed, 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, overall health, and medical history of the patient being treated, as well as similar factors well known in the medical field.

[0178] A physician with ordinary skill in the art, such as an internist or veterinarian, can easily determine and prescribe the effective amount of the required pharmaceutical composition. For example, an internist or veterinarian can start administering the pharmaceutical composition containing the disclosed compound at a level lower than the level required to achieve the desired therapeutic effect, and gradually increase the dose until the desired effect is achieved.

[0179] In specific embodiments, it is particularly advantageous to formulate the compound in dose unit formulations for ease of administration and uniformity of dosage. As used herein, a dose unit formulation refers to a physically separate unit suitable as a unit dose for a patient receiving treatment, each unit containing a predetermined amount of the disclosed compound calculated to produce the desired therapeutic effect, along with the necessary pharmaceutical vehicle. The dose unit formulation is defined and directly dependent on (a) the inherent characteristics of the disclosed compound and the specific therapeutic effect to be achieved, and (b) the inherent limitations in the art of compounding / formulating such disclosed compound for the treatment of a patient's pain, depressive disorder, or drug dependence.

[0180] In one embodiment, the compounds provided herein are 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.

[0181] The optimal ratio, individual doses, and combined doses of drug compounds that produce efficacy without toxicity, as well as their concentrations, are determined using methods known to those skilled in the art, based on the kinetics of the availability of the active ingredient to the target site.

[0182] The routes of administration for any of the compositions considered herein include oral, nasal, rectal, vaginal, parenteral, buccal, sublingual, or topical. Compounds can be formulated for administration by any preferred route, e.g., oral or parenteral, percutaneous, transmucosal (e.g., sublingual, lingual, (trans) buccal, (trans) urethral, ​​vaginal (e.g., transvaginal and perivaginal), (trans) nasal and (trans) rectal), intravesical, intrapulmonary, intraduodenal, intragastric, intrathecal, subcutaneous, intramuscular, intradermal, intra-arterial, intravenous, intrabronchial, inhalation, and topical administration. In one embodiment, the preferred route of administration is oral.

[0183] Suitable compositions and dosage forms include, for example, tablets, capsules, caplets, pills, gelatin capsules, lozenges, dispersants, suspensions, liquids, syrups, granules, beads, transdermal patches, gels, powders, pellets, magma preparations, lozenges, creams, pastes, transdermal patches, lotions, discs, suppositories, liquid sprays for nasal or oral administration, dry powders or aerosol preparations for inhalation, and compositions and preparations for intravesical administration. It should be understood that the preparations and compositions are not limited to the specific preparations and compositions described herein.

[0184] For oral administration, tablets, sugar-coated tablets, liquids, drops, suppositories, or capsules, caplets, and gelatin capsules are particularly preferred. 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 and non-toxic pharmaceutically acceptable excipients suitable for the manufacture of tablets. Such excipients include, for example, inert diluents such as lactose; granulators and disintegrants such as corn starch; binders such as starch; and lubricants such as magnesium stearate. Tablets may or may not be coated, or they may be coated by known techniques for aesthetic purposes or to delay the release of the active ingredient. Oral formulations may also be provided as hard gelatin capsules in which the active ingredient is mixed with an inert diluent.

[0185] For parenteral administration, the disclosed compound can be formulated for injection or infusion, for example, intravenous, intramuscular, or subcutaneous injection or infusion, or for administration by bolus or continuous infusion. Suspensions, solutions, or emulsions in oily or aqueous vehicles may be used and optionally contain other formulation agents such as suspending agents, stabilizers, or dispersants. kit

[0186] In one embodiment, the 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.

[0187] In a particular embodiment, 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.

[0188] In some embodiments, the kit comprises a pharmaceutical composition comprising a MAT2A inhibitor or a pharmaceutically acceptable salt thereof; a topoisomerase inhibitor, splicing inhibitor sulfonamide (SPLAM), or KIF18 inhibitor or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or diluent.

[0189] In additional embodiments, a pharmaceutical kit is provided. The kit includes a sealed container approved for the storage of a pharmaceutical composition, the container containing one of the pharmaceutical compositions described above. In some embodiments, the sealed container minimizes contact between air and the components, for example, an airless bottle. In other embodiments, the sealed container is a sealed tube. The kit shall include instructions for the use of the composition and information about the composition.

[0190] In specific embodiments, the combination of compounds can be administered on the same schedule, whether as a single formulation or unit dosage form containing all of the combination compounds, or as separate formulations or unit dosage forms of the combination compounds. However, some of the compounds used in the combination may be administered more frequently than once a day, or at a different frequency than the other compounds in the combination. Therefore, in one embodiment, the kit contains a formulation or unit dosage form containing all of the compounds in the combination, and an additional formulation or unit dosage form containing one of the compounds in the combination but not the additional active compound, along with instructions for administering the dosage forms on a set schedule.

[0191] The kits provided herein include, for example, prescription information provided to the patient or healthcare provider, or as a label on the packaged pharmaceutical product. Prescription information may include, for example, information on efficacy, dosage and administration, contraindications, and side effects of the pharmaceutical product.

[0192] In all of the foregoing, the combination of compounds of the present invention may be administered alone, as a mixture, or with additional activators.

[0193] The kits provided herein may be designed to accommodate the conditions necessary to properly maintain the components contained therein (e.g., refrigeration or freezing). The kits may include a label or accompanying leaflet containing identification information for the components contained therein, and instructions for use (e.g., administration parameters, clinical pharmacology of the active ingredient(s), e.g., mechanism of action(s), pharmacokinetics and pharmacodynamics, side effects, contraindications, etc.).

[0194] Each component of the kit can be sealed in an individual container, and all of the various containers can be housed in a single package. The label or accompanying information may include manufacturer information such as lot number and expiration date. The label or accompanying information can be, for example, integrated with the physical structure containing the components, housed separately within the physical structure, or affixed to the components of the kit (e.g., ampoules, syringes, or vials).

[0195] Non-exclusive exemplary embodiments: Section A - Non-Limited Illustrative Embodiments 1-112 In the following further embodiments 1 to 112, the disclosure includes the following:

[0196] Embodiment 1. A method is provided for treating cancer in a subject in need thereof, 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.

[0197] Embodiment 2. A method is provided for treating cancer in a subject in need thereof, comprising administering a therapeutically effective amount of a MAT2A inhibitor to the subject, wherein the subject has previously been treated with a KIF18 inhibitor.

[0198] Embodiment 3. In Embodiment 3, there is provided a method of treating cancer in a subject that needs it, the method comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor, wherein the subject is simultaneously receiving administration of a KIF18 inhibitor.

[0199] Embodiment 4. In Embodiment 4, there is provided a method according to any one of Embodiments 1 to 3, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: [Chemical formula] (wherein, X is N or CH, R 3 is C 1-6 haloalkyl, halo, or C 3-6 cycloalkyl, R 2 is -NR 4 R 5 where, R 4 is hydrogen or C 1-6 alkyl, R 5 is hydrogen, C 1-6 alkyl or C 3-6 cycloalkyl, R 1 is phenyl, where the phenyl is substituted with 0 to 2 halos).

[0200] Embodiment 5. In Embodiment 5, there is provided a method according to any one of Embodiments 1 to 4, wherein the KIF18 inhibitor is a KIF18A inhibitor.

[0201] Embodiment 6. In Embodiment 6, there is provided a method according to any one of Embodiments 1 to 5, wherein the KIF18A inhibitor is sobirnesib (Compound B) or a pharmaceutically acceptable salt thereof.

[0202] Embodiment 7. Embodiment 7 provides one of the methods from Embodiments 1 to 6, wherein the MAT2A inhibitor is administered simultaneously or sequentially with the KIF18 inhibitor.

[0203] Embodiment 8. Embodiment 8 provides a method for treating cancer in a subject in need thereof, 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).

[0204] Embodiment 9. Embodiment 9 provides a method for treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor, wherein the subject has previously been treated with a splicing inhibitor sulfonamide (SPLAM).

[0205] Embodiment 10. Embodiment 10 provides a method for treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor, wherein the subject simultaneously receives an administration of a splicing inhibitor sulfonamide (SPLAM).

[0206] Embodiment 11. Embodiment 11 provides one of the methods of Embodiments 8 to 10, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: [ka] (In the formula, X is either N or CH. R 3 C 1-6 Haloalkyl, halo, or C 3-6 It is a cycloalkyl, R 2 -NR 4 R 5 And, R 4is hydrogen or C 1-6 It is alkyl, R 5 is hydrogen, C 1-6 Alkyl or C 3-6 It is a cycloalkyl, R 1 (where phenyl is substituted with 0-2 halos).

[0207] Embodiment 12. Embodiment 12 provides a method according to any one of Embodiments 8 to 11, wherein the SPLAM is indislam or a pharmaceutically acceptable salt thereof.

[0208] Embodiment 13. Embodiment 13 provides a method according to any one of Embodiments 8 to 11, wherein SPLAM is selected from the group consisting of E7820, chloroquinoxaline sulfonamide, and tasislam, or pharmaceutically acceptable salts thereof.

[0209] Embodiment 14. Embodiment 14 provides a method according to any one of Embodiments 8 to 13, wherein the MAT2A inhibitor is administered simultaneously with or sequentially with SPLAM.

[0210] Embodiment 15. Embodiment 15 provides a method for treating cancer in a subject in need thereof, 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.

[0211] Embodiment 16. Embodiment 16 provides a method for treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor, wherein the subject has previously been treated with a topoisomerase inhibitor.

[0212] Embodiment 17. Embodiment 17 provides a method for treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor, wherein the subject simultaneously receives an administration of a topoisomerase inhibitor.

[0213] Embodiment 18. Embodiment 18 provides a method according to any one of Embodiments 15 to 17, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: [ka] (In the formula, X is either N or CH. R 3 C 1-6 Haloalkyl, halo, or C 3-6 It is a cycloalkyl, R 2 -NR 4 R 5 And, R 4 is hydrogen or C 1-6 It is alkyl, R 5 is hydrogen, C 1-6 Alkyl or C 3-6 It is a cycloalkyl, R 1 (where phenyl is substituted with 0-2 halos).

[0214] Embodiment 19. Embodiment 19 provides a method according to any one of Embodiments 15 to 18, wherein the topoisomerase inhibitor is a type I topoisomerase inhibitor.

[0215] Embodiment 20. Embodiment 20 provides the method according to Embodiment 19, wherein the type I topoisomerase inhibitor is selected from the group consisting of 10-hydroxycamptothecin, irinotecan, and topotecan, or pharmaceutically acceptable salts thereof.

[0216] Embodiment 20A. Embodiment 20A provides the method according to Embodiment 19, wherein the type I topoisomerase inhibitor is selected from the group consisting of hexylresorcinol, exatecan, deruxtecan, and berotecan, or pharmaceutically acceptable salts thereof.

[0217] Embodiment 21. Embodiment 21 provides a method according to any one of Embodiments 15 to 18, wherein the topoisomerase inhibitor is a type I topoisomerase inhibitor.

[0218] Embodiment 22. Embodiment 22 provides the method of Embodiment 21, wherein the type II topoisomerase inhibitor is selected from the group consisting of daunorubicin, doxorubicin, and etoposide, or pharmaceutically acceptable salts thereof.

[0219] Embodiment 23. Embodiment 23 provides a method according to any one of Embodiments 15 to 22, wherein the MAT2A inhibitor is administered simultaneously or sequentially with the topoisomerase inhibitor.

[0220] Embodiment 24. Embodiment 24 provides a method according to any one of Embodiments 4-7, 11-14, and 18-23, wherein X is N.

[0221] Embodiment 25. Embodiment 25 provides a method according to any one of Embodiments 4-7, 11-14, and 18-23, wherein X is CH.

[0222] Embodiment 26. In Embodiment 26, R 4 However, it is hydrogen, R 5 However, hydrogen or C 1-3 A method according to any one of embodiments 4-7, 11-14, and 18-25 is provided, wherein the alkyl is used.

[0223] Embodiment 27. In Embodiment 27, R 5A method according to any one of embodiments 4-7, 11-14, and 18-26 is provided, wherein the element is hydrogen or methyl.

[0224] Embodiment 28. In Embodiment 28, R 1 However, a method is provided according to any one of embodiments 4-7, 11-14, and 18-27, wherein the phenyl is substituted with chloro.

[0225] Embodiment 29. In Embodiment 29, R 3 However, C 1-3 A method according to any one of embodiments 4-7, 11-14, and 18-28 is provided, wherein the method is a haloalkyl or a halo.

[0226] Embodiment 30. In Embodiment 30, R 3 A method according to any one of embodiments 4-7, 11-14, and 18-29 is provided, wherein the substance is trifluoromethyl or chloromethyl.

[0227] Embodiment 31. In Embodiment 31, R 3 A method is provided according to any one of embodiments 4-7, 11-14, and 18-28, wherein the substance is cyclopropyl.

[0228] Embodiment 32. Embodiment 32 provides a method according to any one of Embodiments 1 to 31, wherein the MAT2A inhibitor is selected from the group consisting of the compounds in Table 1 or pharmaceutically acceptable salts thereof.

[0229] Embodiment 33. Embodiment 33 provides a method according to any one of Embodiments 1 to 32, wherein the MAT2A inhibitor is compound A or a pharmaceutically acceptable salt thereof.

[0230] Embodiment 34. Embodiment 34 provides a method according to any one of Embodiments 1 to 32, wherein the MAT2A inhibitor is compound A1 or a pharmaceutically acceptable salt thereof.

[0231] Embodiment 35. Embodiment 35 provides a method according to any one of Embodiments 1 to 34, 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, esophageal cancer, malignant peripheral nerve sheath tumor, mesothelioma, salivary gland tumor, urothelial carcinoma, gastrointestinal cancer, and sarcoma.

[0232] Embodiment 36. Embodiment 36 provides a method according to any one of Embodiments 1 to 35, wherein the cancer is a solid tumor or a hematological carcinoma.

[0233] Embodiment 37. Embodiment 37 provides a method according to any one of Embodiments 1 to 36, wherein the cancer is a solid tumor.

[0234] Embodiment 38. Embodiment 38 provides the method according to any one of Embodiments 1 to 37, wherein the cancer is a solid malignant tumor.

[0235] Embodiment 38A. Embodiment 38A provides a method according to any one of Embodiments 1 to 37, wherein the cancer is characterized by decreased or deleted MTAP gene expression, deletion of the MTAP gene, impaired function of the MTAP protein, decreased or deleted levels of the MTAP protein, accumulation of MTA, or a combination thereof.

[0236] Embodiment 39. Embodiment 39 provides one of the methods from Embodiments 1 to 38 and 38A, wherein the cancer is characterized by MTAP gene deletion.

[0237] Embodiment 40. Embodiment 40 provides a use of a MAT2A inhibitor in the manufacture of a pharmaceutical product for treating cancer, wherein the MAT2A inhibitor is administered simultaneously or sequentially with a KIF18 inhibitor.

[0238] Embodiment 41. The KIF18 inhibitor of Embodiment 40, wherein the KIF18 inhibitor in Embodiment 41 is a KIF18A inhibitor.

[0239] Embodiment 42. The KIF18A inhibitor of Embodiment 41, wherein the KIF18A inhibitor in Embodiment 42 is sovilnesib (compound B) or a pharmaceutically acceptable salt thereof.

[0240] Embodiment 43. Embodiment 43 provides a use of a MAT2A inhibitor in the manufacture of a pharmaceutical product for treating cancer, wherein the MAT2A inhibitor is administered simultaneously with or sequentially with SPLAM.

[0241] Embodiment 44. The SPLAM of Embodiment 43, wherein the SPLAM in Embodiment 44 is indislam or a pharmaceutically acceptable salt thereof.

[0242] Embodiment 45. The SPLAM of Embodiment 43, wherein the SPLAM in Embodiment 45 is selected from the group consisting of E7820, chloroquinoxaline sulfonamide, and tasislam, or pharmaceutically acceptable salts thereof.

[0243] Embodiment 46. Embodiment 46 provides a use of a MAT2A inhibitor in the manufacture of a pharmaceutical product for treating cancer, wherein the MAT2A inhibitor is administered simultaneously or sequentially with a topoisomerase inhibitor.

[0244] Embodiment 47. The topoisomerase inhibitor of Embodiment 46, wherein the topoisomerase inhibitor in Embodiment 47 is a type I topoisomerase inhibitor.

[0245] Embodiment 48. The type I topoisomerase inhibitor of Embodiment 47, wherein the type I topoisomerase inhibitor in Embodiment 48 is selected from the group consisting of 10-hydroxycamptothecin, irinotecan, and topotecan, or pharmaceutically acceptable salts thereof.

[0246] Embodiment 48A. Embodiment 48A provides the use described in Embodiment 47, wherein the type I topoisomerase inhibitor is selected from the group consisting of hexylresorcinol, exatecan, deruxtecan, and berotecan, or pharmaceutically acceptable salts thereof.

[0247] Embodiment 49. The topoisomerase inhibitor of Embodiment 48, wherein the topoisomerase inhibitor in Embodiment 49 is a type II topoisomerase inhibitor.

[0248] Embodiment 50. The type II topoisomerase inhibitor of Embodiment 49, wherein the type II topoisomerase inhibitor in Embodiment 50 is selected from the group consisting of daunorubicin, doxorubicin, and etoposide, or pharmaceutically acceptable salts thereof.

[0249] Embodiment 51. Embodiment 51 provides any one of the uses of Embodiments 40 to 50, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: [ka] (In the formula, X is either N or CH. R 3 C 1-6 Haloalkyl, halo, or C 3-6 It is a cycloalkyl, R 2 -NR 4 R 5 And, R 4 is hydrogen or C 1-6 It is alkyl, R 5 is hydrogen, C 1-6 Alkyl or C 3-6 It is a cycloalkyl, R 1 (where phenyl is substituted with 0-2 halos).

[0250] Embodiment 52. Embodiment 52 provides the use of the MAT2A inhibitor of Embodiment 51, wherein the MAT2A inhibitor is a compound of formula (I) as defined in any one of Embodiments 24 to 31 or a pharmaceutically acceptable salt thereof.

[0251] Embodiment 53. Embodiment 53 provides the use of any one of the MAT2A inhibitors from Embodiments 40 to 52, wherein the MAT2A inhibitor is selected from the group consisting of the compounds in Table 1 or pharmaceutically acceptable salts thereof.

[0252] Embodiment 54. Embodiment 54 provides the use of any one of the MAT2A inhibitors from Embodiments 40 to 53, wherein the MAT2A inhibitor is compound A or a pharmaceutically acceptable salt thereof.

[0253] Embodiment 55. Embodiment 55 provides the use of any one of the MAT2A inhibitors from Embodiments 40 to 53, wherein the MAT2A inhibitor is compound A1 or a pharmaceutically acceptable salt thereof.

[0254] Embodiment 56. Embodiment 56 provides a MAT2A inhibitor for use in the treatment of cancer, wherein the MAT2A inhibitor is administered simultaneously with or sequentially with a KIF18 inhibitor.

[0255] Embodiment 57. The KIF18 inhibitor of Embodiment 57, wherein the KIF18 inhibitor is a KIF18A inhibitor.

[0256] Embodiment 58. The KIF18A inhibitor of Embodiment 57, wherein the KIF18A inhibitor in Embodiment 58 is sovilnesib (compound B) or a pharmaceutically acceptable salt thereof.

[0257] Embodiment 59. Embodiment 59 provides a MAT2A inhibitor for use in the treatment of cancer, wherein the MAT2A inhibitor is administered simultaneously with or sequentially with SPLAM.

[0258] Embodiment 60. The SPLAM of Embodiment 59, wherein the SPLAM in Embodiment 60 is indislam or a pharmaceutically acceptable salt thereof.

[0259] Embodiment 61. The SPLAM of Embodiment 59, wherein the SPLAM in Embodiment 61 is selected from the group consisting of E7820, chloroquinoxaline sulfonamide, and tasislam, or pharmaceutically acceptable salts thereof.

[0260] Embodiment 62. Embodiment 62 provides a MAT2A inhibitor for use in the treatment of cancer, wherein the MAT2A inhibitor is administered simultaneously with or sequentially with a topoisomerase inhibitor.

[0261] Embodiment 63. The topoisomerase inhibitor of Embodiment 62, wherein the topoisomerase inhibitor in Embodiment 63 is a type I topoisomerase inhibitor.

[0262] Embodiment 64. The type I topoisomerase inhibitor of Embodiment 63, wherein the type I topoisomerase inhibitor in Embodiment 64 is selected from the group consisting of 10-hydroxycamptothecin, irinotecan, and topotecan, or pharmaceutically acceptable salts thereof.

[0263] Embodiment 64A. Embodiment 64A provides the use described in Embodiment 63, wherein the type I topoisomerase inhibitor is selected from the group consisting of hexylresorcinol, exatecan, deruxtecan, and berotecan, or pharmaceutically acceptable salts thereof.

[0264] Embodiment 65. The topoisomerase inhibitor of Embodiment 62, wherein the topoisomerase inhibitor in Embodiment 65 is a type II topoisomerase inhibitor.

[0265] Embodiment 66. The use of the type II topoisomerase inhibitor of Embodiment 65, wherein the type II topoisomerase inhibitor in Embodiment 66 is selected from the group consisting of daunorubicin, doxorubicin, and etoposide, or pharmaceutically acceptable salts thereof.

[0266] Embodiment 67. Embodiment 67 provides the use of any one of the MAT2A inhibitors from Embodiments 56 to 66, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: [ka] (In the formula, X is either N or CH. R 3 C 1-6 Haloalkyl, halo, or C 3-6 It is a cycloalkyl, R 2 -NR 4 R 5 And, R 4 is hydrogen or C 1-6 It is alkyl, R 5 is hydrogen, C 1-6 Alkyl or C 3-6 It is a cycloalkyl, R 1 (where phenyl is substituted with 0-2 halos).

[0267] Embodiment 68. Embodiment 68 provides the use of the MAT2A inhibitor of Embodiment 67, wherein the MAT2A inhibitor is a compound of formula (I) as defined in any one of Embodiments 24 to 31 or a pharmaceutically acceptable salt thereof.

[0268] Embodiment 69. Embodiment 69 provides the use of any one of the MAT2A inhibitors from Embodiments 56 to 68, wherein the MAT2A inhibitor is selected from the group consisting of the compounds in Table 1 or pharmaceutically acceptable salts thereof.

[0269] Embodiment 70. Embodiment 70 provides the use of any one of the MAT2A inhibitors from Embodiments 56 to 69, wherein the MAT2A inhibitor is compound A or a pharmaceutically acceptable salt thereof.

[0270] Embodiment 71. Embodiment 71 provides the use of any one of the MAT2A inhibitors from Embodiments 56 to 69, wherein the MAT2A inhibitor is compound A1 or a pharmaceutically acceptable salt thereof.

[0271] Embodiment 72. Embodiment 72 provides the use described in any one of Embodiments 40 to 71, wherein the cancer is selected from the group consisting of leukemia, glioma, lung 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.

[0272] Embodiment 73. Embodiment 73 provides the use according to any one of Embodiments 40 to 72, wherein the cancer is a solid tumor or a hematological carcinoma.

[0273] Embodiment 74. Embodiment 74 provides the use according to any one of Embodiments 40 to 73, wherein the cancer is a solid tumor.

[0274] Embodiment 75. Embodiment 75 provides the use according to any one of Embodiments 40 to 74, wherein the cancer is a solid malignant tumor.

[0275] Embodiment 76. Embodiment 76 provides the use described in any one of Embodiments 40 to 75, wherein the cancer is characterized by decreased or deleted MTAP gene expression, deletion of the MTAP gene, impaired function of the MTAP protein, decreased or deleted levels of the MTAP protein, accumulation of MTA, or a combination thereof.

[0276] Embodiment 77. Embodiment 77 provides one of the methods from Embodiments 40 to 76, wherein the cancer is characterized by MTAP gene deletion.

[0277] Embodiment 78. Embodiment 78 provides a method for inhibiting tumor growth or slowing the growth rate of a tumor in a subject with MTAP-deficient cancer, the method comprising administering a MAT2A inhibitor and a KIF18 inhibitor, SPLAM, or a topoisomerase inhibitor.

[0278] Embodiment 79. Embodiment 79 provides the method of Embodiment 78, wherein tumor growth is measured by the change in tumor volume from a first time point to a second time point.

[0279] Embodiment 80. Embodiment 80 provides the method of Embodiment 79, wherein the tumor volume at the second time point has not increased compared to the first time point.

[0280] Embodiment 81. Embodiment 81 provides the method of Embodiment 80, wherein the tumor volume decreases from the first time point to the second time point.

[0281] Embodiment 82. Embodiment 82 provides a method for treating cancer in a subject in need thereof, comprising administering to the subject a MAT2A inhibitor and a KIF18 inhibitor, SPLAM, or a topoisomerase inhibitor, wherein the subject has previously received a cancer treatment regimen that does not include a MAT2A inhibitor.

[0282] Embodiment 83. Embodiment 83 provides the method of Embodiment 82, wherein treatment with the MAT2A inhibitor and a KIF18 inhibitor, SPLAM, or a topoisomerase inhibitor reduces the tumor growth rate compared to treatment with the MAT2A inhibitor alone for the same period.

[0283] Embodiment 84. Embodiment 84 provides a combination product comprising a MAT2A inhibitor and an inhibitor selected from the group consisting of a KIF18 inhibitor, SPLAM, and a topoisomerase inhibitor.

[0284] Embodiment 85. Embodiment 85 provides a combination product comprising a first pharmaceutical composition containing a therapeutically effective amount of a MAT2A inhibitor and a second pharmaceutical composition containing a therapeutically effective amount of an inhibitor selected from the group consisting of a KIF18 inhibitor, SPLAM, and a topoisomerase inhibitor.

[0285] Embodiment 86. In Embodiment 86, the MAT2A inhibitor of Embodiment 84 or 85 is a compound of formula (I) or a pharmaceutically acceptable salt thereof: [ka] (In the formula, X is either N or CH. R 3 C 1-6 Haloalkyl, halo, or C 3-6 It is a cycloalkyl, R 2 -NR 4 R 5 And, R 4 is hydrogen or C 1-6 It is alkyl, R 5 is hydrogen, C 1-6 Alkyl or C 3-6 It is a cycloalkyl, R 1 (where phenyl is substituted with 0-2 halos).

[0286] Embodiment 87. In Embodiment 87, the MAT2A inhibitor of Embodiment 86 is a compound of formula (I) as defined in any one of Embodiments 24 to 31 or a pharmaceutically acceptable salt thereof.

[0287] Embodiment 88. In Embodiment 88, one of the MAT2A inhibitors from Embodiments 84 to 87 is selected from the group consisting of the compounds in Table 1 or their pharmaceutically acceptable salts.

[0288] Embodiment 89. In Embodiment 89, one of the MAT2A inhibitors from Embodiments 84 to 88 is compound A or a pharmaceutically acceptable salt thereof.

[0289] Embodiment 90. In Embodiment 90, one of the MAT2A inhibitors from Embodiments 84 to 88 is compound A1 or a pharmaceutically acceptable salt thereof.

[0290] Embodiment 91. In Embodiment 91, one of the KIF18 inhibitors from Embodiments 84 to 90 is a KIF18A inhibitor.

[0291] Embodiment 92. The KIF18A inhibitor of Embodiment 91, wherein the KIF18A inhibitor in Embodiment 92 is sovilnesib (compound B) or a pharmaceutically acceptable salt thereof.

[0292] Embodiment 93. An embodiment 93 in which the SPLAM is any one of embodiments 84 to 90, wherein the SPLAM is indislam or a pharmaceutically acceptable salt thereof.

[0293] Embodiment 94. In Embodiment 94, the SPLAM is selected from the group consisting of E7820, chloroquinoxaline sulfonamide, and tasislam, or pharmaceutically acceptable salts thereof, one of the SPLAMs of Embodiments 84 to 90.

[0294] Embodiment 95. A topoisomerase inhibitor according to any one of Embodiments 84 to 90, wherein the topoisomerase inhibitor in Embodiment 95 is a type I topoisomerase inhibitor.

[0295] Embodiment 96. The type I topoisomerase inhibitor of Embodiment 95, wherein the type I topoisomerase inhibitor in Embodiment 96 is selected from the group consisting of 10-hydroxycamptothecin, irinotecan, and topotecan, or pharmaceutically acceptable salts thereof.

[0296] Embodiment 96A. The type I topoisomerase inhibitor of Embodiment 95, wherein the type I topoisomerase inhibitor in Embodiment 96A is selected from the group consisting of hexylresorcinol, exatecan, deruxtecan, and berotecan, or pharmaceutically acceptable salts thereof.

[0297] Embodiment 96B. The type I topoisomerase inhibitor of Embodiment 95, wherein the type I topoisomerase inhibitor in Embodiment 96B is irinotecan.

[0298] Embodiment 97. A topoisomerase inhibitor according to any one of Embodiments 84 to 90, wherein the topoisomerase inhibitor in Embodiment 97 is a type II topoisomerase inhibitor.

[0299] Embodiment 98. The use of the type II topoisomerase inhibitor of Embodiment 97, wherein the type II topoisomerase inhibitor in Embodiment 98 is selected from the group consisting of daunorubicin, doxorubicin, and etoposide, or pharmaceutically acceptable salts thereof.

[0300] Embodiment 99. Embodiment 99 provides a method for treating cancer in a subject in need thereof, 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.

[0301] Embodiment 100. In Embodiment 100, the ADC containing the topoisomerase inhibitor is a type I topoisomerase inhibitor.

[0302] Embodiment 101. In Embodiment 101, the ADC containing the topoisomerase inhibitor is fam-trastuzumab deruxtecan-nxki.

[0303] Embodiment 102. In Embodiment 102, the ADC containing the topoisomerase inhibitor is AZD8205.

[0304] Embodiment 103. In Embodiment 103, the ADC containing the topoisomerase inhibitor is DS-1062.

[0305] Embodiment 104. In Embodiment 104, one of the MAT2A inhibitors from Embodiments 99 to 103 is selected from the group consisting of the compounds in Table 1 or their pharmaceutically acceptable salts.

[0306] Embodiment 105. In Embodiment 105, one of the MAT2A inhibitors from Embodiments 99 to 104 is compound A or a pharmaceutically acceptable salt thereof.

[0307] Embodiment 106. In Embodiment 106, one of the MAT2A inhibitors from Embodiments 99 to 104 is compound A1 or a pharmaceutically acceptable salt thereof.

[0308] Embodiment 107. Embodiment 107 provides a method according to any one of Embodiments 99 to 106, wherein the cancer is selected from the group consisting of leukemia, glioma, lung 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.

[0309] Embodiment 108. Embodiment 108 provides a method according to any one of Embodiments 99 to 107, wherein the cancer is a solid tumor or a hematological carcinoma.

[0310] Embodiment 109. Embodiment 109 provides a method according to any one of Embodiments 99 to 108, wherein the cancer is a solid tumor.

[0311] Embodiment 110. Embodiment 110 provides the method according to any one of Embodiments 99 to 109, wherein the cancer is a solid malignant tumor.

[0312] Embodiment 111. Embodiment 111 provides a method according to any one of Embodiments 99 to 110, wherein the cancer is characterized by decreased or deleted MTAP gene expression, deletion of the MTAP gene, impaired function of the MTAP protein, decreased or deleted levels of the MTAP protein, accumulation of MTA, or a combination thereof.

[0313] Embodiment 112. Embodiment 112 provides one of the methods from Embodiments 1 to 111, wherein the cancer is characterized by a deletion of the MTAP gene. Where references to prior embodiments are made in relation to the embodiments described above, such references also include embodiments having literal or combined notations. For example, references to embodiments 40–50 include embodiments 40, 41, 42, 43, 44, 45, 46, 47, 48, 48A, 49, and 50. Section B – Additional Non-Limited Exemplary Embodiments 1–40

[0314] Embodiment 1. A method is provided for treating cancer in a subject in need thereof, 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: [ka] (In the formula, X is either N or CH. R 3 C 1-6 Haloalkyl, halo, or C3-6 It is a cycloalkyl, R 2 -NR 4 R 5 And, R 4 is hydrogen or C 1-6 It is alkyl, R 5 is hydrogen, C 1-6 Alkyl or C 3-6 It is a cycloalkyl, R 1 (where phenyl is substituted with 0-2 halos).

[0315] Embodiment 2. Embodiment 2 provides the method according to Embodiment 1, wherein the topoisomerase inhibitor is a type I topoisomerase inhibitor.

[0316] Embodiment 3. Embodiment 3 provides the method according to Embodiment 2, wherein the type I topoisomerase inhibitor is selected from the group consisting of 10-hydroxycamptothecin, irinotecan, and topotecan, or pharmaceutically acceptable salts thereof.

[0317] Embodiment 4. Embodiment 4 provides the method according to Embodiment 1, wherein the topoisomerase inhibitor is a type II topoisomerase inhibitor.

[0318] Embodiment 5. Embodiment 5 provides the method of Embodiment 4, wherein the type II topoisomerase inhibitor is selected from the group consisting of daunorubicin, doxorubicin, and etoposide, or pharmaceutically acceptable salts thereof.

[0319] Embodiment 6. Embodiment 6 provides one of Embodiments 1 to 5, wherein the MAT2A inhibitor is administered simultaneously or sequentially with the topoisomerase inhibitor.

[0320] Embodiment 7. A method is provided for treating cancer in a subject in need thereof, comprising administering a MAT2A inhibitor to the subject and administering a therapeutically effective amount of a splicing inhibitor sulfonamide (SPLAM) to the subject, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof: [ka] (In the formula, X is either N or CH. R 3 C 1-6 Haloalkyl, halo, or C 3-6 It is a cycloalkyl, R 2 -NR 4 R 5 And, R 4 is hydrogen or C 1-6 It is alkyl, R 5 is hydrogen, C 1-6 Alkyl or C 3-6 It is a cycloalkyl, R 1 (where phenyl is substituted with 0-2 halos).

[0321] Embodiment 8. Embodiment 8 provides the method of Embodiment 7, wherein SPLAM is indislam or a pharmaceutically acceptable salt thereof.

[0322] Embodiment 9. Embodiment 9 provides the method of Embodiment 7, wherein SPLAM is selected from the group consisting of E7820, chloroquinoxaline sulfonamide, and tasislam, or pharmaceutically acceptable salts thereof.

[0323] Embodiment 10. Embodiment 10 provides a method according to any one of Embodiments 7 to 9, wherein the MAT2A inhibitor is administered simultaneously with or sequentially with SPLAM.

[0324] Embodiment 11. Embodiment 11 provides a method for treating cancer in a subject in need thereof, 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: [ka] (In the formula, X is either N or CH. R 3 C 1-6 Haloalkyl, halo, or C 3-6 It is a cycloalkyl, R 2 -NR 4 R 5 And, R 4 is hydrogen or C 1-6 It is alkyl, R 5 is hydrogen, C 1-6 Alkyl or C 3-6 It is a cycloalkyl, R 1 (where phenyl is substituted with 0-2 halos).

[0325] Embodiment 12. Embodiment 12 provides the method according to Embodiment 11, wherein the KIF18 inhibitor is a KIF18A inhibitor.

[0326] Embodiment 13. Embodiment 13 provides the method of Embodiment 12, wherein the KIF18A inhibitor is sovilnesib or a pharmaceutically acceptable salt thereof.

[0327] Embodiment 14. Embodiment 14 provides one of the methods from Embodiments 11B to 13B, in which the MAT2A inhibitor is administered simultaneously or sequentially with the KIF18 inhibitor.

[0328] Embodiment 15. Embodiment 15 provides a method according to any one of Embodiments 1 to 14, wherein X is N.

[0329] Embodiment 16. Embodiment 16 provides a method according to any one of Embodiments 1 to 14, wherein X is CH.

[0330] Embodiment 17. In Embodiment 17, R 4 However, it is hydrogen, R 5 However, hydrogen or C 1-3 A method according to any one of embodiments 1 to 16 is provided, wherein the alkyl is used.

[0331] Embodiment 18. In Embodiment 18, R 5 A method according to any one of Embodiments 1 to 17 is provided, wherein the hydrogen is or methyl.

[0332] Embodiment 19. In Embodiment 19, R 1 However, a method according to any one of Embodiments 1 to 18 is provided, wherein the phenyl is substituted with chloro.

[0333] Embodiment 20. In Embodiment 20, R 3 However, C 1-3 A method according to any one of Embodiments 1 to 19 is provided, wherein the material is a haloalkyl or a halo.

[0334] Embodiment 21. In Embodiment 21, R 3 A method according to any one of Embodiments 1 to 20 is provided, wherein the trifluoromethyl or chloromethyl is used.

[0335] Embodiment 22. In Embodiment 22, R 3 A method according to any one of Embodiments 1 to 20 is provided, wherein the material is cyclopropyl.

[0336] Embodiment 23. Embodiment 23 provides a method according to any one of Embodiments 1 to 22, wherein the MAT2A inhibitor is selected from the group consisting of the compounds in Table 1 or pharmaceutically acceptable salts thereof.

[0337] Embodiment 24. In Embodiment 24, the MAT2A inhibitor is compound A: [ka] A method according to any one of Embodiments 1 to 23 is provided, which is either a pharmaceutically acceptable salt thereof.

[0338] Embodiment 25. In Embodiment 25, the MAT2A inhibitor is compound A1: [ka] A method according to any one of Embodiments 1 to 23 is provided, which is either a pharmaceutically acceptable salt thereof.

[0339] Embodiment 26. Embodiment 26 provides a method according to any one of Embodiments 1 to 25, 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, esophageal cancer, malignant peripheral nerve sheath tumor, mesothelioma, salivary gland tumor, urothelial carcinoma, gastrointestinal cancer, and sarcoma.

[0340] Embodiment 27. Embodiment 27 provides a method according to any one of Embodiments 1 to 26, wherein the cancer is a solid tumor or a hematological carcinoma.

[0341] Embodiment 28. Embodiment 28 provides a method according to any one of Embodiments 1 to 27, wherein the cancer is a solid malignant tumor.

[0342] Embodiment 29. Embodiment 29 provides a method according to any one of Embodiments 1 to 28, wherein the cancer is characterized by decreased or deleted MTAP gene expression, deletion of the MTAP gene, impaired function of the MTAP protein, decreased or deleted levels of the MTAP protein, accumulation of MTA, or a combination thereof.

[0343] Embodiment 30. Embodiment 30 provides the use of a MAT2A inhibitor in the manufacture of a pharmaceutical product for treating cancer, wherein the MAT2A inhibitor is administered together with a topoisomerase inhibitor.

[0344] Embodiment 31. Embodiment 31 provides the use of a MAT2A inhibitor in the manufacture of a pharmaceutical product for treating cancer, wherein the MAT2A inhibitor is administered together with SPLAM.

[0345] Embodiment 32. Embodiment 32 provides the use of a MAT2A inhibitor in the manufacture of a pharmaceutical product for treating cancer, wherein the MAT2A inhibitor is administered together with a KIF18 inhibitor.

[0346] Embodiment 33. Embodiment 33 provides the use of any one of Embodiments 30 to 32, wherein the MAT2A inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0347] Embodiment 34. Embodiment 34 provides the use of Embodiment 33B, wherein the MAT2A inhibitor is compound A or a pharmaceutically acceptable salt thereof, or compound A1 or a pharmaceutically acceptable salt thereof.

[0348] Embodiment 35. Embodiment 35 provides a method for inhibiting tumor growth or slowing the growth rate of a tumor in a subject with MTAP-deficient cancer, the method comprising administering a MAT2A inhibitor and a topoisomerase inhibitor, SPLAM, or a KIF18 inhibitor.

[0349] Embodiment 36. Embodiment 36 provides the method of Embodiment 35, wherein tumor growth is measured by the change in tumor volume from a first time point to a second time point.

[0350] Embodiment 37. Embodiment 37 provides the method of Embodiment 36, wherein the tumor volume at the second time point has not increased compared to the first time point.

[0351] Embodiment 38. Embodiment 38 provides the method of Embodiment 37, wherein the tumor volume decreases from the first time point to the second time point.

[0352] Embodiment 39. Embodiment 39 provides a method for treating cancer in a subject in need thereof, comprising administering to the subject a MAT2A inhibitor and a topoisomerase inhibitor, SPLAM, or KIF18 inhibitor, wherein the subject has previously received a cancer treatment regimen that does not include a MAT2A inhibitor.

[0353] Embodiment 40. Embodiment 40 provides the method of Embodiment 39, wherein treatment with the MAT2A inhibitor and a topoisomerase inhibitor, SPLAM, or KIF18 inhibitor reduces the tumor growth rate compared to treatment with the MAT2A inhibitor alone for the same period.

[0354] Those skilled in the art will recognize, or can verify, many equivalents to the specific procedures, embodiments, claims, and examples described herein without performing any experiments beyond the usual procedures. Such equivalents are deemed to be within the scope of this disclosure and included in the appended claims. For example, it should be understood that modifying reaction conditions, such as, but not limited to, reaction time, reaction size / volume, and experimental reagents, such as solvents, catalysts, pressure, atmospheric conditions, such as nitrogen atmosphere, and reducing / oxidizing agents, using substitutes recognized in the art without performing any experiments beyond the usual procedures is within the scope of this application.

[0355] Where values ​​and ranges are provided herein, it should be understood that all values ​​and ranges encompassed by these values ​​and ranges are included within the scope of this disclosure. Furthermore, all values ​​within these ranges, as well as any upper or lower limits on the ranges of values, are also contemplated in this application.

[0356] The following embodiments further illustrate aspects of the present disclosure. However, these embodiments do not limit in any way the teachings of the present disclosure described herein. [Examples]

[0357] The compounds and methods disclosed herein will be further illustrated by the following examples, but should not be construed as further limitations. Unless otherwise specified, the implementation of this disclosure will employ the prior art of organic synthesis, cell biology, cell culture, and molecular biology within the scope of the art.

[0358] Example 1: Synergistic growth inhibition in vitro by equimolar combination of MAT2A inhibitor and KIF18A inhibitor. Materials and methods A 10-day growth assay was performed on a panel of five MTAP-deficient cancer cell lines and two wild-type cell lines of non-small cell lung cancer and bladder cancer. Optimal cell seeding for all cell lines was determined by evaluating growth at various seeding densities in a 384-well format to identify conditions that enabled growth over 10 days. Cells were then plated at the optimal seeding density and treated with a 20-point 2-fold dilution series of compound A, sovilnesib ("compound B"), or equimolar combinations of compound A and compound B. The concentrations tested for compound A and compound B individually or in combination ranged from 0.038 nM to 20,000 nM. Cell plates were collected upon compound addition, and the cell count at the start of the combination (T0) was quantified. To quantify the cells, the collected cells were lysed using Promega CellTiter-Glo (CTG) reagent according to the manufacturer's protocol, and the chemiluminescence signal was detected using a Synergy Neo plate reader. CTG estimates cell number by detecting intracellular ATP levels. Cells were incubated with a drug combination at 37°C and 5% CO2 for 10 days. Subsequently, the cells were lysed with CTG and the chemiluminescence signal was measured.

[0359] analysis The CTG values ​​obtained after 10 days of treatment were subtracted from the background and expressed as a percentage of the T0 value. The data were plotted against compound concentration and fitted with a four-parameter equation to create dose-response curves for each single compound and equimolar combination. Growth ICx (gICx) values ​​in the range of gIC30 to gIC100 were interpolated from the fitted curves. Synergistic growth inhibition was evaluated by determining the combination index (CI) values ​​at various points throughout the titration using the non-mutually exclusive equation shown below (where A corresponds to the gIC value of compound A, B corresponds to the gICx value of compound B, and the values ​​of A+B or B+A correspond to the gICx values ​​of the equimolar combination) [Chou, 1983; Chou, 1981]. A combination index value less than 1.0 was considered synergistic.

number

[0360] result Synergistic growth inhibition was observed in MTAP-deficient cell lines with a combined effect of compound A and compound B at multiple gICx points in combined titration (Tables 3A-3E). The synergistic effect of the combination was also observed in wild-type cell lines with a combined effect of 1 / 2 (Tables 4A-4B). Although considered synergistic, the combined index value in wild-type TCCSUP cell lines was ≥0.7. Compared to wild-type cell lines, MTAP-deficient cell lines showed higher sensitivity to the combination, with a low combined index value of 0.01.

[0361] Table 3A-3E: Synergistic growth inhibition by equimolar combinations of compound A and compound B across five MTAP-deficient cell lines. For equimolar combinations of compound A and compound B, the synergistic effect in gICx values ​​in the range of gIC30 to gIC100 was measured using a non-mutually exclusive combination index formula. The combination index values ​​for NCI-H838(3A), SW900(3B), RT112 / 84(3C), UMUC5(3D), and UMUC11(3E) are shown. [Table 2] [Table 3] [Table 4] [Table 5] [Table 6]

[0362] Tables 4A-4B: Synergistic growth inhibition by equimolar combinations of compound A and compound B across two wild-type cell lines. For equimolar combinations of compound A and compound B, the synergistic effect in gICx values ​​ranging from gIC30 to gIC100 was measured using a non-mutually exclusive combination index formula. The combination index values ​​for NCI-H520(4A) and TCCSUP(4B) are shown. [Table 7] [Table 8]

[0363] Example 2: Synergistic growth inhibition in vitro by combining a KIF18A inhibitor and a fixed-concentration MAT2A inhibitor. Materials and methods A 10-day growth assay was performed on a panel of five MTAP-deficient cancer cell lines and two wild-type cell lines of non-small cell lung cancer and bladder cancer. Optimal cell seeding for all cell lines was determined by evaluating growth at various seeding densities in a 384-well format to identify conditions that enabled growth over 10 days. Cells were then plated at the optimal seeding density and treated with a 20-point 2-fold dilution series of compound B in combination with a fixed concentration of compound A at 1000 nM. For comparison, single-agent titrations of compound A and compound B were also included. The concentrations tested for compound A and compound B individually or in combination ranged from 0.038 nM to 20,000 nM. Cell plates were collected upon compound addition, and the cell count at the start of the combination (T0) was quantified. To quantify the cells, the collected cells were lysed using Promega CellTiter-Glo (CTG) reagent according to the manufacturer's protocol, and the chemiluminescence signal was detected as described in Example 1 above. Cells were incubated with drug combinations at 37°C and 5% CO2 for 10 days. Subsequently, the cells were lysed with CTG and the chemiluminescence signal was measured.

[0364] analysis The CTG values ​​obtained after 10 days of treatment were subtracted for background and expressed as a percentage with the T0 value set to 100%. The data were plotted against compound concentration and fitted with a 4-parameter equation to create monotherapy dose-response curves. The inhibition percentage for each combination and monotherapy concentration compared to the DMSO control was determined. In addition, the overall net growth or death of cells was calculated and reported as a Growth-Death Index (GDI) value on a scale of -100 to 100. The midpoint (0) represents the cell number at the start of the combination (T0).

[0365] For each titration point of compound B, synergistic growth inhibition in combination with 1000 nM compound A was evaluated using the Bliss independence model. The growth inhibition observed at each point was compared to the predicted inhibition based on the additive activity of each individual compound. The predicted inhibition was calculated using (Ea + Eb) - (Ea * Eb) (where E corresponds to the effect (inhibition) of each individual compound (a and b)). A difference of more than 10% between the observed inhibition rate and the predicted inhibition rate was considered synergistic. Bliss scores were calculated only for combinations that induced growth inhibition greater than 20% (GDI value < 80). Since Bliss is calculated based on growth inhibition, scores within the cytotoxic dose range may not accurately reflect synergistic cytotoxicity.

[0366] result Synergistic growth inhibition was observed in 5 / 5 MTAP-deficient cell lines across numerous titration points with the combination of compound B and 1000 nM compound A, as assessed by Bliss independence (Tables 5A-5E). Due to the steep dose-response curve observed with compound B monotherapy, the range of observable combined synergy within this assay design was narrowed in some cell lines. For example, 5000 nM compound B monotherapy was sufficient to maximally inhibit cell growth in the UMUC5 cell line (Figures 1A-3). Therefore, the dose range in which combined synergy can be observed in UMUC5 is less than 5000 nM of compound B. Despite this limited range, combined synergy was observed in 5 / 5 MTAP-deficient cell lines at doses below the dose required for maximal inhibition by the monotherapy. In these cell lines, synergistic growth inhibition was observed across a wide dose range of the combination of compound B and 1000 nM compound A.

[0367] Enhanced growth inhibition by combination was observed in half wild-type cell lines within a limited dose range of 39–156 nM of compound B (Tables 6A–6B). While a synergistic effect was calculated by Bliss independence in these cell lines, the shift in potency in the dose-response curve was not as significant as that observed in MTAP-deficient cell lines. In Figures 1A-1–1A-5 and 1B-1–1B-2, growth inhibition for compound B alone at titration doses is shown as a percentage relative to the DMSO control. Dose-response curves were prepared using four-parameter fitting for five MTAP-deficient cell lines (Figures 1A-1–1A-5) and two wild-type cell lines (Figures 1B-1 and 1B-2).

[0368] Tables 5A-5E: Synergistic growth inhibition by combination of compound B and 1000 nM compound A across five MTAP-deficient cell lines. The synergistic effect of compound B with compound A at 1000 nM was measured by Bliss independence analysis. Bliss scores greater than 10 are indicated by "*" and are considered synergistic. The Bliss score is valid for combinations showing greater than 20% growth inhibition (GDI < 80). Synergistic effects reported using Bliss may be underestimated within the cytotoxic dose range. The growth death index is shown on a scale of -100 to 100 for combined titrations and for each monochemical compound A (compound A) and compound B (compound B), with the midpoint (0) being the cell number at the start of the combination (T0). Bliss and GDI values ​​are shown for five MTAP-deficient cell lines: NCI-H838 (5A), SW900 (5B), RT112 / 84 (5C), UMUC5 (5D), and UMUC11 (5E). [Table 9] [Table 10] [Table 11] [Table 12] [Table 13]

[0369] Tables 6A-6B: Synergistic growth inhibition by a combination of compound B and 1000 nM compound A across two wild-type cell lines. The synergistic effect of compound A and compound B across titration volumes of 1000 nM compound A was measured by Bliss independence analysis. Bliss scores greater than 10 are indicated with an asterisk (*) and are considered synergistic. Bliss scores were calculated only for combinations showing more than 20% growth inhibition. Synergistic effects reported using Bliss may be underestimated within the cytotoxic dose range. Growth death index values ​​are shown on a scale of -100 to 100 for combined titrations and for each individual compound A (compound A) and compound B (compound B), with the midpoint (0) being the cell number at the start of the combination (T0). Bliss and GDI values ​​for five MTAP-deficient cell lines: NCI-H520 (6A), TCCSUP (6B) are shown. [Table 14] [Table 15]

[0370] Example 3: Identification of combined effects of MAT2A inhibition and indisram by proliferation screening. Materials and methods Fifteen pancreatic, non-small cell lung, and bladder cancer cell lines (10 MTAP nulls and 5 MTAP wild-types) underwent a 10-day growth screening. Optimal cell seeding for all cell lines was determined by evaluating growth at various seeding densities in a 384-well format to identify conditions that enabled growth over 10 days. Cells were then plated at the optimal seeding density in the presence of 20–150 nM of compound A or a DMSO vehicle control. Cells were incubated at 37°C and 5% CO2 for 4 days to enable target engagement of the pretreatment compounds. Maintaining the pretreatment conditions, cells were further treated with an 11-point 3x titration series of compounds obtained from a chemically diverse library of 424 compounds. Combination compound concentrations ranged from 0.2 nM to 14,679 nM. Cell plates were collected upon addition of the combination compounds, and the cell count at the start of the combination (T0) was quantified. To quantify the cells, the harvested cells were lysed with CellTiter-Glo (CTG) (Promega) reagent according to the manufacturer's protocol, and the chemiluminescence signal was detected using a Synergy Neo plate reader (ThermoFisher, serial number 140715A). CTG estimates cell number by detecting intracellular ATP levels. The cells were incubated with the drug combination at 37°C and 5% CO2 for a further 6 days, completing a total assay period of 10 days including pretreatment. Subsequently, the cells were lysed with CTG, and the chemiluminescence signal was measured. The CTG values ​​obtained after 10 days of treatment were subtracted from the background, expressed as a percentage of the T0 value, and plotted against the compound concentration. The data were fitted to a 4-parameter equation to create concentration-response curves. For each combination titration, growth IC50 values ​​and maximum growth inhibition were compared between DMSO-pretreated cells and compound A-pretreated cells. Combination hits were determined based on observations of a shift in growth IC50 greater than 2x and / or a reduction in growth inhibition greater than 20%.

[0371] result 424 compounds were tested in combination with compound A in 10 MTAP null cell lines and 5 MTAP wild-type cell lines. Of these compounds, indislam was identified as the most consistent combination hit in the screening, and compound A pretreatment resulted in a more than twofold shift in growth IC50 in 7 out of 10 MTAP-deficient cell lines and 1 out of 5 wild-type cell lines (Table 7).

[0372] Table 7: Growth ICs in 10 cancer cell lines with and without pretreatment of compound A 50 value Growth IC of Indislam and Compound A 50 And the values ​​of the magnification change are shown. 2x or more growth IC 50 A magnification shift (indicated by "*") was observed in 7 / 10 MTAP-deficient and 1 / 5 wild-type cell lines. NCI-H838 cell lines were tested with two concentrations of compound A (20 nM and 150 nM). In both of these conditions, growth IC50 was greater than 2x. 50 A scaling shift in the values ​​was observed. [Table 16]

[0373] Example 4: Synergistic in vitro growth inhibition by equimolar combination of a MAT2A inhibitor and four structurally different splicing inhibitor sulfonamides. Materials and methods A 10-day growth assay was performed on a panel of five MTAP-deficient cancer cell lines and two wild-type cell lines of non-small cell lung cancer and bladder cancer. Optimal cell seeding for all cell lines was determined by evaluating growth at various seeding densities in a 384-well format to identify conditions that enabled growth over 10 days. Cells were then plated at the optimal seeding density and treated with a 20-point 2-fold dilution series of compound A, splicing inhibitor sulfonamide (SPLAM), or equimolar combinations of compound A and SPLAM. Combinations tested included compound A with four structurally distinct SPLAMs. The concentrations tested for compound A and each SPLAM, individually or in combination, ranged from 0.04 nM to 20,000 nM. Cell plates were collected upon compound addition, and the cell count at the start of each combination (T0) was quantified. To quantify the cells, the harvested cells were lysed using Promega CellTiter-Glo (CTG) reagent according to the manufacturer's protocol, and the chemiluminescence signal was detected using a Synergy Neo plate reader. CTG estimates cell number by detecting intracellular ATP levels. The cells were incubated with drug combinations at 37°C and 5% CO2 for 10 days. Subsequently, the cells were lysed with CTG, and the chemiluminescence signal was measured.

[0374] analysis The CTG values ​​obtained after 10 days of treatment were subtracted from the background and expressed as a percentage of the T0 value. The data were plotted against compound concentration and fitted with a four-parameter equation to create dose-response curves for each single compound and equimolar combination. Growth ICx (gICx) values ​​in the range of gIC30 to gIC100 were interpolated from the fitted curves. Synergistic growth inhibition was evaluated by determining the combination index (CI) values ​​at various points throughout the titration using the non-mutually exclusive equation shown below (where A corresponds to the gICx value of compound A, B corresponds to the gICx value of SPLAM, and the values ​​of A+B or B+A correspond to the gICx values ​​of the equimolar combination) [Chou, 1983; Chou, 1981]. A combination index value less than 1.0 was considered synergistic.

number

[0375] result Synergistic growth inhibition was observed in a panel of MTAP-deficient cell lines at multiple gICx points of combined titration with compound A and four different SPLAMs (Tables 8A-8E, 9A-9E, 10A-10E, 11A-11E, and 12A-12E). Combination synergies were also observed in two wild-type cell lines, particularly with the more potent SPLAMs, Indislam and E7820 (Tables 8F-8G, 9F-9G, 10F-10G, and 11F-11G). Although considered synergistic, the combination index values ​​in wild-type cell lines were higher than those observed in MTAP-deficient cell lines. Compared to wild-type cell lines, MTAP-deficient cell lines showed greater sensitivity to the combinations and greater growth inhibition.

[0376] Tables 8A-8G: Synergistic growth inhibition by equimolar combinations of compound A and indislam across five MTAP-deficient cell lines and two wild-type cell lines. For equimolar combinations of compound A and indislam, the synergistic effect on gICx values ​​in the range of gIC30 to gIC100 was measured using a non-mutually exclusive combination index formula. The combination index values ​​for MTAP-deficient cell lines (8A-8E) and wild-type cell lines (8F-8G) are shown. [Table 17] [Table 18] [Table 19] [Table 20] [Table 21] [Table 22] [Table 23]

[0377] Tables 9A-9G: Synergistic growth inhibition by equimolar combinations of compound A and E7820 across five MTAP-deficient cell lines and two wild-type cell lines. For equimolar combinations of compound A and E7820, the synergistic effect on gICx values ​​in the range of gIC30 to gIC100 was measured using a non-mutually exclusive combination index formula. The combination index values ​​for MTAP-deficient cell lines (9A-9E) and wild-type cell lines (9F-9G) are shown. [Table 24] [Table 25] [Table 26] [Table 27] [Table 28] [Table 29] [Table 30]

[0378] Tables 10A-10G: Synergistic growth inhibition by equimolar combinations of compound A and chloroquinoxaline sulfonamide across five MTAP-deficient cell lines and two wild-type cell lines. For equimolar combinations of compound A and chloroquinoxaline sulfonamide, the synergistic effect on gICx values ​​in the range of gIC30 to gIC100 was measured using a non-mutually exclusive combination index formula. The combination index values ​​for MTAP-deficient cell lines (10A to 10E) and wild-type cell lines (10F to 10G) are shown. [Table 31] [Table 32] [Table 33] [Table 34] [Table 35] [Table 36] [Table 37]

[0379] Tables 11A-11G: Synergistic growth inhibition by equimolar combinations of compound A and tasislam across five MTAP-deficient cell lines and two wild-type cell lines. For equimolar combinations of compound A and tasislam, the synergistic effect on gICx values ​​in the range of gIC30 to gIC100 was measured using a non-mutually exclusive combination index formula. The combination index values ​​for MTAP-deficient cell lines (11A to 11E) and wild-type cell lines (11F to 11G) are shown. [Table 38] [Table 39] [Table 40] [Table 41] [Table 42] [Table 43] [Table 44]

[0380] Tables 12A-12E: Synergistic growth inhibition by indislam and 1000 nM compound A across five MTAP-deficient cell lines. The synergistic effect of indislam with 1000 nM compound A was measured by Bliss independence analysis across titration volumes. Bliss scores greater than 10 are indicated by "*" and are considered synergistic. The Bliss score is valid for combinations showing greater than 20% growth inhibition (GDI < 80). Synergistic effects reported using Bliss may be underestimated within the cytotoxic dose range. The growth-kill index is shown on a scale of -100 to 100 for combined titrations and for each monochemical compound A (compound A) and indislam, with the midpoint (0) being the cell number at the start of the combination (T0). Scores less than 0 are indicated by "**" and represent a cytotoxic response. The Bliss and GDI values ​​for five MTAP-deficient cell lines: NCI-H838(12A), SW900(12B), RT112 / 84(12C), UMUC5(12D), and UMUC11(12E) are shown. [Table 45] [Table 46] [Table 47] [Table 48] [Table 49]

[0381] Example 5: Synergistic growth inhibition in vitro by a combination of four splicing inhibitor sulfonamides and a fixed concentration MAT2A inhibitor. Materials and methods A 10-day growth assay was performed on a panel of five MTAP-deficient cancer cell lines and two wild-type cell lines of non-small cell lung cancer and bladder cancer. Optimal cell seeding for all cell lines was determined by evaluating growth at various seeding densities in a 384-well format to identify conditions that enabled growth over 10 days. Cells were then plated at the optimal seeding density and treated with each of four structurally different SPLAMs in a 20-point 2-fold dilution series in combination with a fixed concentration of compound A at 1000 nM. For comparison, single-agent titrations of compound A and each SPLAM were also included. The concentrations tested for each SPLAM, either alone or in combination, ranged from 0.04 nM to 20,000 nM. Cell plates were collected upon compound addition, and the cell count at the start of the combination (T0) was quantified. To quantify the cells, the collected 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 were incubated with drug combinations at 37°C and 5% CO2 for 10 days. Subsequently, the cells were lysed with CTG and the chemiluminescence signal was measured.

[0382] analysis The CTG values ​​obtained after 10 days of treatment were subtracted for background and expressed as a percentage with the T0 value set to 100%. The data were plotted against compound concentration and fitted with a 4-parameter equation to create monotherapy dose-response curves. The inhibition percentage for each combination and monotherapy concentration compared to the DMSO control was determined. In addition, the overall net growth or death of cells was calculated and reported as a Growth-Death Index (GDI) value on a scale of -100 to 100. The midpoint (0) represents the cell number at the start of the combination (T0).

[0383] For each titration point of SPLAM, synergistic growth inhibition in combination with 1000 nM compound A was evaluated using the Bliss independence model. The growth inhibition observed at each point was compared to the predicted inhibition based on the additive activity of each mono-agent dose. The predicted inhibition was calculated using (Ea + Eb) - (Ea * Eb) (where E corresponds to the effect (inhibition) of each mono-agent (a and b)). A difference of more than 10% between the observed inhibition rate and the predicted inhibition rate was considered synergistic. Bliss scores were reported only for combinations that induced growth inhibition greater than 20% (GDI value < 80). Since Bliss is calculated based on growth inhibition, scores within the cytotoxic dose range may not accurately reflect synergistic cytotoxicity.

[0384] result Synergistic growth inhibition was observed in 7 / 7 cell lines across numerous titration points for each of the four structurally distinct SPLAMs combined with 1000 nM compound A, as assessed by Bliss independence (Tables 13A–13B, 14A–14E, 15A–15B, 16A–16E, 17A–17B, 18A–18E, and 19A–19B). Within the context of each SPLAM, greater growth inhibition and synergistic effects were observed in MTAP-deficient cell lines at lower SPLAM doses than in wild-type cell lines. Combination cytotoxicity was observed only in the MTAP-deficient NCI-H838, SW900, and RT112 / 84 cell lines, and not in any wild-type cell lines.

[0385] Tables 13A-13B: Synergistic growth inhibition by a combination of indislam and 1000 nM compound A across two wild-type cell lines. The synergistic effect of indislam with 1000 nM compound A was measured by Bliss independence analysis. Bliss scores greater than 10 are indicated with an asterisk (*) and are considered synergistic. The Bliss score is valid for combinations showing greater than 20% growth inhibition (GDI < 80). Synergistic effects reported using Bliss may be underestimated within the cytotoxic dose range. The growth-kill index values ​​are shown on a scale of -100 to 100 for combined titrations and for each monochemical compound A (compound A) and indislam, with the midpoint (0) being 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: NCI-H520 (13A), TCCSUP (13B) are shown. [Table 50] [Table 51]

[0386] Tables 14A-14E: Synergistic growth inhibition by combination of E7820 and 1000 nM compound A across five MTAP-deficient cell lines. The synergistic effect of E7820 with a 1000 nM compound A was measured by Bliss independence analysis. A Bliss score greater than 10 is indicated by "*" and is considered synergistic. The Bliss score is valid for combinations showing more than 20% growth inhibition (GDI < 80). Synergistic effects reported using Bliss may be underestimated within the cytotoxic dose range. The growth-kill index is shown on a scale of -100 to 100 for the combined titration and for each individual compound A (compound A) and indislam, with the midpoint (0) being the cell number at the start of the combination (T0). A score less than 0 is indicated by "**" and represents a cytotoxic response. The Bliss and GDI values ​​for five MTAP-deficient cell lines: NCI-H838(14A), SW900(14B), RT112 / 84(14C), UMUC5(14D), and UMUC11(14E) are shown. [Table 52] [Table 53] [Table 54] [Table 55] [Table 56]

[0387] Tables 15A-15B: Synergistic growth inhibition by a combination of E7820 and 1000 nM compound A across two wild-type cell lines. The synergistic effect of E7820 with 1000 nM compound A across titration volumes was measured by Bliss independence analysis. Bliss scores greater than 10 are indicated by "*" and are considered synergistic. The Bliss score is valid for combinations showing greater than 20% growth inhibition (GDI < 80). Synergistic effects reported using Bliss may be underestimated within the cytotoxic dose range. The growth-kill index values ​​are shown on a scale of -100 to 100 for combined titrations and for each monochemical compound A (compound A) and indislam, with the midpoint (0) being the cell number at the start of the combination (T0). Scores less than 0 are indicated by "**" and represent a cytotoxic response. Bliss and GDI values ​​for five MTAP-deficient cell lines (NCI-H520 (15A), TCCSUP (15B)) are shown. [Table 57] [Table 58]

[0388] Tables 16A-16E: Synergistic growth inhibition by a combination of chloroquinoxalinesulfonamide and 1000 nM compound A across five MTAP-deficient cell lines. The synergistic effect of chloroquinoxaline sulfonamide with 1000 nM compound A was measured by independent analysis. A Bliss score greater than 10 is indicated by "*" and is considered synergistic. The Bliss score is valid for combinations showing growth inhibition greater than 20% (GDI < 80). Synergistic effects reported using Bliss may be underestimated within the cytotoxic dose range. The growth-kill index is shown on a scale of -100 to 100 for combined titrations and for each monochemical compound A (compound A) and indislam, with the midpoint (0) being the cell number at the start of the combination (T0). A score less than 0 is indicated by "**" and represents a cytotoxic response. The Bliss and GDI values ​​for five MTAP-deficient cell lines: NCI-H838(16A), SW900(16B), RT112 / 84(16C), UMUC5(16D), and UMUC11(16E) are shown. [Table 59] [Table 60] [Table 61] [Table 62] [Table 63]

[0389] Tables 17A-17B: Synergistic growth inhibition by a combination of chloroquinoxalinesulfonamide and 1000 nM compound A across two wild-type cell lines. The synergistic effect of chloroquinoxaline sulfonamide with 1000 nM compound A was measured by independent analysis. Bliss scores greater than 10 are indicated by "*" and are considered synergistic. The Bliss score is valid for combinations showing greater than 20% growth inhibition (GDI < 80). Synergistic effects reported using Bliss may be underestimated within the cytotoxic dose range. The growth-kill index is shown on a scale of -100 to 100 for combined titrations and for each monochemical compound A (compound A) and indislam, with the midpoint (0) being the cell number at the start of the combination (T0). Scores less than 0 are indicated by "**" and represent a cytotoxic response. Bliss and GDI values ​​for five MTAP-deficient cell lines: NCI-H520 (17A), TCCSUP (17B) are shown. [Table 64] [Table 65]

[0390] Tables 18A-18E: Synergistic growth inhibition by the combination of tasislam and 1000 nM compound A across five MTAP-deficient cell lines. The synergistic effect of tasislam with 1000 nM compound A across titration volumes was measured by Bliss independence analysis. Bliss scores greater than 10 are indicated by "*" and are considered synergistic. The Bliss score is valid for combinations showing greater than 20% growth inhibition (GDI < 80). Synergistic effects reported using Bliss may be underestimated within the cytotoxic dose range. The growth-kill index is shown on a scale of -100 to 100 for combined titrations and for each monochemical compound A (compound A) and indislam, with the midpoint (0) being the cell number at the start of the combination (T0). Scores less than 0 are indicated by "**" and represent a cytotoxic response. The Bliss and GDI values ​​for five MTAP-deficient cell lines: NCI-H838(18A), SW900(18B), RT112 / 84(18C), UMUC5(18D), and UMUC11(18E) are shown. [Table 66] [Table 67] [Table 68] [Table 69] [Table 70]

[0391] Tables 19A-19B: Synergistic growth inhibition by a combination of tasislam and 1000 nM compound A across two wild-type cell lines. The synergistic effect of tasislam with 1000 nM compound A across titration volumes was measured by Bliss independence analysis. Bliss scores greater than 10 are indicated by "*" and are considered synergistic. The Bliss score is valid for combinations showing greater than 20% growth inhibition (GDI < 80). Synergistic effects reported using Bliss may be underestimated within the cytotoxic dose range. The growth-kill index is shown on a scale of -100 to 100 for combined titrations and for each monotherapy agent (compound A) and tasislam, with the midpoint (0) being the cell number at the start of the combination (T0). Scores less than 0 are indicated by "**" and represent a cytotoxic response. Bliss and GDI values ​​for five MTAP-deficient cell lines: NCI-H520 (19A), TCCSUP (19B) are shown. [Table 71] [Table 72]

[0392] Example 6: MAT2A and topoisomerase inhibitors produce a combined effect in an in vitro MTAP deficiency model. Materials and Methods A panel of 13 MTAP-deficient 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 was used to evaluate the combinatorial effects of a MAT2A inhibitor (Compound A) and topoisomerase inhibitors: 10-hydroxycamptothecin, irinotecan, topotecan, daunorubicin, doxorubicin, and etoposide. For combinatorial screening, cells were seeded in 384-well plates at 150 cells / well for Compound A or 500 cells / well for topoisomerase inhibitors to determine the optimal concentration range of each compound in each cell line. After 24 hours, cells were treated as single agents with each compound in 9-point titrations and incubated at 37 °C, 5% CO2 for 6 days. For combinatorial screening, cells were seeded in 384-well plates at 150 cells / well. After 24 hours, cells were co-treated with each compound in 5-point titrations in an optimized 6×6 dose matrix and incubated at 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) for quantification of cell number. For quantification of cell number using CTG, plates of untreated cells were harvested at the time of compound addition (T0 or zero time point). Each data point was performed in technical triplicates.

[0393] The percent growth inhibition was calculated as follows: When T < V0: 100*(1 - (T - V0) / V0) When T ≥ V0: 100*(1 - (T - V0) / (V - V0))

[0394] where T is the signal measurement of the test substance, V is the measurement of the vehicle-treated control, and V0 is the measurement of the vehicle control at T0. The percent growth inhibition was used to generate dose-response curves for single drug activity and drug combination synergy using Chalice Analyzer software (Horizon) and GI 50This was used in the calculation. Growth inhibition of 100% and growth inhibition exceeding 100% indicate cell arrest and cytotoxicity, respectively.

[0395] Synergistic growth inhibition was evaluated using the Loewe additiveity model. The growth inhibition observed at each dose was compared to the predicted inhibition based on the additive activity of each drug when combined with itself. Loewe additiveity was calculated as (X / X I )+(Y / Y I I that satisfy )=1 Loewe And here, X I and Y I This represents the effective concentration of the single agent required to obtain the observed combination effect I. A synergistic effect was considered to exist when the difference between the observed and predicted growth inhibition was 20% or greater.

[0396] The synergy score was calculated to quantify the strength of the synergy, as follows: Synergistic effect score = log f X log f Y Σ Maximum (0,I データ )(I データ -I Loewe )

[0397] The inhibition rates for each component drug and combination point in the matrix were calculated relative to the median of all untreated / vehicle-treated control wells. A synergistic effect was considered to have occurred if the synergistic effect score was greater than 2.22. This corresponds to effects that show high growth inhibition at multiple dose points and exceed 20% in the Loewe additive model.

[0398] result Combinations of compound A with six different topoisomerase inhibitors were tested in a panel of 13 MTAP-deficient cell lines. Synergistic growth inhibition was observed with each combination, but the degree varied across the cell line panel. For example, the combination of compound A with 10-hydroxycamptothecin increased the observed growth inhibition compared to either compound alone and showed a strong synergistic effect (determined by a synergy score greater than 2.22) in 10 / 13 MTAP-deficient cell lines (Figures 2A-1 to 2G, 3A to 3G-2, and Table 20).

[0399] The combination of compound A and irinotecan also showed increased growth inhibition compared to the single agent and demonstrated a synergistic effect in 8 / 13 MTAP-deficient cell lines (Figures 4A-1 to 4G, 5A to 5G-2, and Table 20). The combination of compound A and topotecan also increased the growth inhibition observed with either single agent and demonstrated a synergistic effect in 6 / 13 MTAP-deficient cell lines (Figures 6A-1 to 6G, 7A to 7G-2, and Table 20). The combination of compound A and daunorubicin showed increased growth inhibition and a strong synergistic effect in 12 / 13 MTAP-deficient cell lines (Figures 8A-1 to 8G, 9A to 9G-2, and Table 20). The combination of compound A and doxorubicin showed increased growth inhibition and a strong synergistic effect in MTAP-deficient cell lines (8 / 13) (Figures 10A-1 to 10G, 11A to 11G-2, and Table 20). The combination of compound A and etoposide showed increased growth inhibition and a strong synergistic effect in MTAP-deficient cell lines (5 / 13) (Figures 12A-1 to 12G, 13A to 13G-2, and Table 20).

[0400] In the growth inhibition diagram, growth inhibition is shown as a percentage of T0 for each cell line in a 6x6 dose matrix. Growth inhibition of 50-100% is indicated by "*", and 100% growth inhibition represents cell arrest. Growth inhibition greater than 100% is highlighted in dark gray and represents cytotoxicity. In the Loewe synergy count, synergies are measured at the total test concentrations of compound A and 10-hydroxycamptothecin using the Loewe additive model for each cell line. Values ​​greater than 20 are indicated by "**" and are considered synergistic. [Table 73]

[0401] Example 7: Antitumor activity of compound A and irinotecan in the RT112 / 84 mouse xenograft model All mouse studies were conducted in accordance with National Institutes of Health (NIH) guidelines and approved by the Institutional Animal Care and Use Committee (IACUC). Mice were kept in pathogen-free conditions and given free access to food and water.

[0402] The antitumor effects of compound A and irinotecan hydrochloride, either alone or in combination, were evaluated in a xenograft (CDX) model derived from RT112 / 84 human bladder tumor cells. Cells were cultured in RPMI-1640 (Gibco, catalog no. 11875093) containing 10% fetal bovine serum. These cells were mycoplasma-free and verified by STR profiling. Five million cells in the logarithmic growth phase were resuspended in Hanks equilibrium salt solution containing 50% Matrigel and subcutaneously transplanted into the flanks of female Crl:NU-Foxn1nu mice. Mice were housed in micro-isolator cages lined with corn cobs. The cages were enriched with sterile nesting material (Innovive) and bio-hat (Bio-Serv). Water (Innovive) and diet (Teklad Global 19% Protein Extruded Diet 2919, irradiated) were provided ad libitum. The environment was maintained at approximately 68-72°F and 40-60% relative humidity with a 12-hour lighting cycle.

[0403] The tumor volume (TV) is calculated using the following formula: TV(mm 3 The value was calculated using ) = (short diameter × short diameter × long diameter) / 2. Tumor growth inhibition (TGI) was measured as [(control group TV 最終 - Treatment Group TV 最終 ) / (Control group TV 最終 - Control group TV 開始時 The TV was calculated using [(x) × 100]. Statistical significance of TV was analyzed using GraphPad Prism version 10.0.3. Repeated measures two-way ANOVA with Tukey's multiple comparisons was used to obtain a P-value from data collected on day 14 (end day for the vehicle group), and a value of less than 0.05 was considered statistically significant. Tumor regression was calculated using the percentage change in tumor volume when comparing the final tumor volume measurement on day 73 with the baseline tumor volume on day 1. Tumor regression was defined as a decrease in TV on day 73 compared to day 1. The tumor regression rate was defined as the percentage of animals in each group that showed tumor regression.

[0404] The average tumor volume at the start of treatment was approximately 249 mm². 3 Ten mice were randomly assigned to each treatment group. The mice were administered either a vehicle, compound A 10 mg / kg orally once daily, compound B 5 mg / kg intraperitoneally (IP) for three consecutive days followed by a four-day rest period, or a combination of compound A and compound B. The vehicle group consisted of a combination of vehicle A (0.5% 400 cps methylcellulose and 0.5% Tween-80 sterile water in the case of compound A) and vehicle B (physiological saline in the case of irinotecan hydrochloride).

[0405] result Treatment with compound A alone resulted in a 53% TGI rate, while irinotecan hydrochloride alone resulted in an 83% TGI rate. The combination of compound A and irinotecan hydrochloride resulted in a 90% TGI rate, as shown in Table 21 and Figure 14. The combination of compound A and irinotecan hydrochloride increased the number of mice with tumor regression, as shown in Table 21 and Figure 15, and achieved a 50% response rate at day 73. In contrast, irinotecan hydrochloride alone resulted in a 30% response rate. [Table 74]

[0406] Example 8: Antitumor activity of compound A and irinotecan in the gastric cancer mouse xenograft model MKN45 All mouse studies were conducted in accordance with National Institutes of Health (NIH) guidelines and approved by the Institutional Animal Care and Use Committee (IACUC). Mice were kept in pathogen-free conditions and given free access to food and water.

[0407] The antitumor effects of compound A and irinotecan hydrochloride, either alone or in combination, were evaluated in a xenograft (CDX) model derived from MKN45 (RCB1001) human gastric tumor cells. MKN45 cells were cultured in RPMI1640 containing 10% fetal bovine serum. The cells were mycoplasma-free and verified by STR profiling. Ten million cells in the logarithmic growth phase were resuspended in Hanks equilibrium salt solution containing 50% Matrigel and subcutaneously transplanted into the flanks of female Crl:NU-Foxn1nu mice. The mice were housed in micro-isolator cages lined with corn cobs. The cages were enriched with sterile nesting material (Innovive) and bio-hat (Bio-Serv). Water (Innovive) and diet (Teklad Global 19% Protein Extruded Diet 2919, irradiated) were provided ad libitum. The environment was maintained at approximately 68-72°F and 40-60% relative humidity with a 12-hour lighting cycle.

[0408] The tumor volume (TV) is calculated using the following formula: TV(mm 3 The value was calculated using ) = (short diameter × short diameter × long diameter) / 2. Tumor growth inhibition (TGI) was measured as [(control group TV 最終 - Treatment Group TV 最終 ) / (Control group TV 最終 - Control group TV 開始時 The results were calculated using [(x) × 100]. Statistical significance was analyzed using GraphPad Prism version 10.2.2. Repeated measures two-way ANOVA with Dunnet's multiple comparisons was used to obtain p-values ​​from data collected on the vehicle group's final day. A p-value of less than 0.05 was considered statistically significant.

[0409] The average tumor volume at the start of treatment was approximately 190 mm². 3Six mice were randomly assigned to each treatment group. The mice received either a vehicle, compound A 10 mg / kg orally once daily, compound B 5 mg / kg or 10 mg / kg administered intraperitoneally (IP) for three consecutive days followed by a four-day rest period, or a combination of compound A and compound B. The vehicle group consisted of a combination of vehicle A (0.5% 400 cps methylcellulose and 0.5% Tween-80 sterile water in the case of compound A) and vehicle B (physiological saline in the case of irinotecan hydrochloride).

[0410] result Treatment with compound A alone resulted in a 56% TGI, while irinotecan hydrochloride alone resulted in a TGI of 49%–85%. The combination of compound A and irinotecan hydrochloride resulted in a 103% TGI, as shown in Table 22 and Figure 16. The combination of compound A and irinotecan hydrochloride increased the number of mice with tumor regression, as shown in Table 22, and achieved an 83% tumor regression rate on day 25. In contrast, irinotecan hydrochloride alone resulted in a regression rate of 0–17%. [Table 75]

[0411] Example 9: Antitumor activity of compound A and irinotecan in LMSU, a mouse xenograft model of gastric cancer All mouse studies were conducted in accordance with National Institutes of Health (NIH) guidelines and approved by the Institutional Animal Care and Use Committee (IACUC). Mice were kept in pathogen-free conditions and given free access to food and water.

[0412] The antitumor effects of compound A and irinotecan hydrochloride, either alone or in combination, were evaluated in an LMSU (RCB1062) human gastric tumor cell-derived xenograft (CDX) model. LMSU cells were cultured in HamF10 containing 10% fetal bovine serum. Cells were mycoplasma-free and verified by STR profiling. Ten million cells in the logarithmic growth phase were resuspended in Hanks equilibrium salt solution containing 50% Matrigel and subcutaneously transplanted into the flanks of female Crl:NU-Foxn1nu mice. Mice were housed in micro-isolator cages lined with corn cobs. The cages were enriched with sterile nesting material (Innovive) and bio-hat (Bio-Serv). Water (Innovive) and diet (Teklad Global 19% Protein Extruded Diet 2919, irradiated) were provided ad libitum. The environment was maintained at approximately 68-72°F and 40-60% relative humidity with a 12-hour lighting cycle.

[0413] The tumor volume (TV) is calculated using the following formula: TV(mm 3 The value was calculated using ) = (short diameter × short diameter × long diameter) / 2. Tumor growth inhibition (TGI) was measured as [(control group TV 最終 - Treatment Group TV 最終 ) / (Control group TV 最終 - Control group TV 開始時 The results were calculated using [(x) × 100]. Statistical significance was analyzed using GraphPad Prism version 10.2.2. Repeated measures two-way ANOVA with Dunnet's multiple comparisons was used to obtain p-values ​​from data collected on the vehicle group's final day. A p-value of less than 0.05 was considered statistically significant.

[0414] The average tumor volume at the start of treatment was approximately 185 mm². 3Seven mice were randomly assigned to each treatment group. The mice received either a vehicle, compound A 10 mg / kg orally once daily, compound B 5 mg / kg or 10 mg / kg administered intraperitoneally (IP) for three consecutive days followed by a four-day rest period, or a combination of compound A and compound B. The vehicle group consisted of a combination of vehicle A (0.5% 400 cps methylcellulose and 0.5% Tween-80 sterile water in the case of compound A) and vehicle B (physiological saline in the case of irinotecan hydrochloride).

[0415] result Treatment with compound A alone did not result in significant tumor growth induction (TGI), but irinotecan hydrochloride alone produced TGI in 42%–80% of mice. The combination of compound A and irinotecan hydrochloride produced TGI in 97% of mice, as shown in Table 23 and Figure 17. The combination of compound A and irinotecan hydrochloride increased the number of mice with tumor regression, as shown in Table 23, and a tumor regression rate of 29% was achieved on day 25. In contrast, irinotecan hydrochloride alone produced a regression rate of 0–14%. [Table 76]

[0416] This specification describes specific embodiments of the Invention, including the best mode for carrying out the Invention as recognized by the inventors. It is assumed that, by reading the foregoing description, variations of the embodiments of this disclosure will become apparent to those skilled in the art, and that such variations may be adopted as appropriate. Therefore, if the Invention is carried out in a manner other than that specifically described herein, it is intended to include all subject matter modifications and equivalents enumerated in the appended claims, as permitted by applicable law. Furthermore, any combination of the elements described above in all possible variations thereof is incorporated herein unless otherwise specifically designated herein or unless it is clearly inconsistent with the context.

[0417] All patent applications, patents, and printed publications referenced herein are incorporated herein by reference in their entirety, except for any definitions, denials or waivers of subject matter, and unless the incorporated material conflicts with the express disclosure herein, in which case the language of this disclosure shall prevail.

[0418] Other embodiments are included in the following claims.

Claims

1. A method for treating cancer in a subject in need thereof, 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, the method: 【Chemistry 1】 (In the formula, X is either N or CH, R ３ C １－６ Haloalkyl, halo, or C ３－６ It is a cycloalkyl, R ２ -NR ４ R ５ And, R ４ is hydrogen or C １－６ It is alkyl, R ５ is hydrogen, C １－６ alkyl or C ３－６ cycloalkyl, and R １ (This is a phenyl molecule substituted with 0 to 2 halos.)

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, exatecan, deruxtecan, berotecan, and topotecan, or pharmaceutically acceptable salts thereof.

4. The method according to 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 according to 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 with or sequentially with the topoisomerase inhibitor.

8. A method for treating cancer in a subject in need thereof, 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, the method: 【Chemistry 2】 (In the formula, X is either N or CH, R ３ C １－６ Haloalkyl, halo, or C ３－６ It is a cycloalkyl, R ２ -NR ４ R ５ And, R ４ is hydrogen or C １－６ It is alkyl, R ５ is hydrogen, C １－６ Alkyl or C ３－６ It is a cycloalkyl, R １ (This is a phenyl molecule substituted with 0 to 2 halos.)

9. The method according to claim 8, wherein the SPLAM is indisram or a pharmaceutically acceptable salt thereof.

10. The method according to claim 8, wherein the SPLAM is selected from the group consisting of E7820, chloroquinoxaline sulfonamide, and tasislam, or pharmaceutically acceptable salts thereof.

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

12. A method for treating cancer in a subject in need thereof, 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, the method: 【Transformation 3】 (In the formula, X is either N or CH, R ３ C １－６ Haloalkyl, halo, or C ３－６ It is a cycloalkyl, R ２ -NR ４ R ５ And, R ４ is hydrogen or C １－６ It is alkyl, R ５ is hydrogen, C １－６ Alkyl or C ３－６ It is a cycloalkyl, R １ (This is a phenyl molecule substituted with 0 to 2 halos.)

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

14. The method according to claim 13, wherein the KIF18A inhibitor is sovilnesib or a pharmaceutically acceptable salt thereof.

15. The method according to any one of claims 12-14, wherein the MAT2A inhibitor is administered simultaneously with or sequentially with the KIF18 inhibitor.

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. R ４ However, it is hydrogen, R ５ However, hydrogen or C １－３ The method according to any one of claims 1-17, wherein the alkyl group is alkyl.

19. R ５ The method according to any one of claims 1 to 18, wherein the is hydrogen or methyl.

20. R １ The method according to any one of claims 1 to 19, wherein the phenyl is substituted with chloro.

21. R ３ However, C １－３ The method according to any one of claims 1-20, wherein the member is a haloalkyl or a halo.

22. R ３ The method according to any one of claims 1 to 21, wherein the substance is trifluoromethyl or chloro.

23. R ３ The method according to any one of claims 1 to 21, wherein the material 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 the compounds in Table 1 or pharmaceutically acceptable salts thereof.

25. The MAT2A inhibitor is compound A: 【Chemistry 4】 The method according to any one of claims 1 to 24, or a pharmaceutically acceptable salt thereof.

26. The MAT2A inhibitor is compound A1: 【Transformation 5】 The method according to any one of claims 1 to 24, or a pharmaceutically acceptable salt thereof.

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, stomach 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, digestive tract cancer, and sarcoma.

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

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 decreased or deleted MTAP gene expression, deletion of the MTAP gene, impaired function of the MTAP protein, decreased or deleted levels of the MTAP protein, accumulation of MTA, or a combination thereof.

31. The use of a MAT2A inhibitor in the manufacture of a pharmaceutical product for the treatment of cancer, wherein the MAT2A inhibitor is used in combination with a topoisomerase inhibitor.

32. The use of a MAT2A inhibitor in the manufacture of a pharmaceutical product for the treatment of cancer, wherein the MAT2A inhibitor is used in combination with SPLAM.

33. The use of a MAT2A inhibitor in the manufacture of a pharmaceutical product for treating cancer, wherein the MAT2A inhibitor is used in combination with a KIF18 inhibitor.

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

35. The use according to 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 the growth rate of a tumor in a subject with MTAP-deficient cancer, 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 according to claim 36, wherein the tumor growth is measured by the change in tumor volume from a first time point to a second time point.

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

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

40. A method for treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a MAT2A inhibitor and a therapeutically effective amount of a topoisomerase inhibitor, SPLAM, or KIF18 inhibitor. The subject is the method, which has not previously received treatment with a MAT2A inhibitor.

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

Images