Cancer treatment using MTA-cooperative PRMT5 inhibitors

Abstract

Described herein are methods for treating cancer in a patient, the methods comprising administering to the patient a PRMT5 inhibitor and a PARP inhibitor, a KRAS inhibitor, or a kinase-like protein 18A (KIF18A) inhibitor, or a kinase inhibitor.

Description

[Background technology]

[0001] Epigenetic regulation of gene expression is an important biological determinant of protein production and cell differentiation and plays a prominent etiological role in several human diseases. Epigenetic regulation involves heritable modifications that do not involve changes in the nucleotide sequence of genetic material. Typically, epigenetic regulation is mediated by selective and reversible modifications (e.g., methylation) of DNA and proteins (e.g., histones) that control the conformational transition of chromatin between transcriptionally active and transcriptionally inactive states. Such covalent modifications can be regulated by enzymes such as methyltransferases (e.g., PRMT5), many of which are associated with specific genetic alterations that can cause human disease. PRMT5 plays a role in diseases such as proliferative, metabolic, and hematological disorders.

[0002] Homozygous deletion of tumor suppressor genes is a key driver of cancer and frequently results in the concomitant loss of passenger genes located in close genomic proximity to the tumor suppressor. Such passenger gene deletions can create tumor-cell-specific, therapeutically tractable vulnerabilities. Homozygous deletions of the chromosome 9p21 locus harboring the well-known tumor suppressor CDKN2A (cyclin-dependent kinase inhibitor 2A) are found in 15% of all tumors and frequently involve the passenger gene MTAP (methylthioadenosine phosphorylase), a key enzyme in the methionine and adenine salvage pathway. Loss of MTAP leads to the accumulation of its substrate, methylthioadenosine (MTA). MTA shares close structural similarity with S-adenosylmethionine (SAM), the methyl donor substrate for the type II methyltransferase PRMT5. Elevated MTA levels, driven by MTAP loss, selectively compete with SAM for binding to PRMT5, placing the methyltransferase in a hypomorphic state that is vulnerable to further PRMT5 inhibition. Multiple genome-wide shRNA dropout screens performed on a large panel of tumor cell lines further emphasized the strength of this metabolic vulnerability by identifying a strong correlation between MTAP loss and cell line PRMT5 dependence. However, PRMT5 is a known cellular essential gene, and conditional PRMT5 knockout and siRNA knockdown studies suggest that significant morbidity may be associated with PRMT5 inhibition in normal tissues (e.g., pancytopenia, infertility, skeletal muscle loss, cardiac hypertrophy, etc.). Therefore, novel strategies that exploit this metabolic vulnerability to preferentially target PRMT5 in MTAP-null tumors while sparing PRMT5 in normal tissues (MTAP WT) are needed. Targeting PRMT5 with MTA-cooperative small-molecule inhibitors may offer an improved therapeutic index compared with normal cells with intact MTAP and low MTA levels, while preferentially targeting the MTA-bound form of PRMT5 that is abundant in MTAP-null tumor cells. Summary of the Invention [Means for solving the problem]

[0003] The present disclosure provides a method of treating cancer in a patient in need thereof, comprising administering to the patient: (a) an amount ranging from 40 mg to 2000 mg of a PRMT5 inhibitor having the formula (I) or the structure [ka] or a pharmaceutically acceptable salt thereof; [ka] (In the ceremony X 1 is NH, N(C1-C6 alkyl), O, or S; X 2 is N(C1-C6 alkyl), O, or S; Y 2 is H, C1-C6 alkyl, or C1-C6 haloalkyl; Z 1 and Z 2 each is independently H, F, or C1-C6 alkyl; and Z 3 , Z 4 , Z 5 , and Z 6 each of which is independently H, C1-C6 alkyl, or chloride; and (b) a second therapeutic agent selected from a PARP inhibitor, a KRAS inhibitor, a kinase-like protein 18A (KIF18A) inhibitor, or a kinase inhibitor. The method includes administering [Brief explanation of the drawings]

[0004] [Figure 1] 1 is a graph showing that the combination of Compound G and sotorasib resulted in significant antitumor activity in LU99 NSCLC xenografts compared to either single agent alone. [Figure 2]1 is a graph showing that the combination of Compound B and sotorasib produced significant antitumor activity in LU99 NSCLC xenografts compared to either single agent alone. [Figure 3] 1 is a graph showing that the combination of Compound B and sotorasib produced significant anti-tumor activity in LU5268 NSCLC patient-derived xenografts compared to either agent alone. [Figure 4] 1 is a graph showing that the combination of Compound B and sotorasib resulted in a decrease in pancreatic cancer (MIAPACA2) cell viability. [Figure 5] 1 is a graph showing that the combination of Compound B and sotorasib significantly reduced the number of pancreatic cancer (MIAPACA2) cells. [Figure 6A] These results demonstrate that the combination of Compound G and sotorasib significantly inhibited tumor growth in MTAP-null, KRAS G12C-mutant NSCLC and PDAC xenografts. Figure 6A: Mice were implanted with LU99 (NSCLC CDX) tumors; Figure 6B: LU5268 (NSCLC PDX) tumors; and Figure 6C: MiaPaCa2 (PDAC CDX) tumors. Vehicle, Compound G, and sotorasib were administered at the indicated doses. Data represent mean ± SD, n = 10 per group. Statistics: P values ​​were determined by linear mixed-effects model and Tukey's overall group comparison: combination vs. either single agent; ****p < 0.0001. [Figure 6B] These results demonstrate that the combination of Compound G and sotorasib significantly inhibited tumor growth in MTAP-null, KRAS G12C-mutant NSCLC and PDAC xenografts. Figure 6A: Mice were implanted with LU99 (NSCLC CDX) tumors; Figure 6B: LU5268 (NSCLC PDX) tumors; and Figure 6C: MiaPaCa2 (PDAC CDX) tumors. Vehicle, Compound G, and sotorasib were administered at the indicated doses. Data represent mean ± SD, n = 10 per group. Statistics: P values ​​were determined by linear mixed-effects model and Tukey's overall group comparison: combination vs. either single agent; ****p < 0.0001. [Figure 6C]These results demonstrate that the combination of Compound G and sotorasib significantly inhibited tumor growth in MTAP-null, KRAS G12C-mutant NSCLC and PDAC xenografts. Figure 6A: Mice were implanted with LU99 (NSCLC CDX) tumors; Figure 6B: LU5268 (NSCLC PDX) tumors; and Figure 6C: MiaPaCa2 (PDAC CDX) tumors. Vehicle, Compound G, and sotorasib were administered at the indicated doses. Data represent mean ± SD, n = 10 per group. Statistics: P values ​​were determined by linear mixed-effects model and Tukey's overall group comparison: combination vs. either single agent; ****p < 0.0001. DETAILED DESCRIPTION OF THE INVENTION

[0005] The present disclosure provides a method of treating cancer in a patient in need thereof, comprising administering to the patient: (a) an amount ranging from 40 mg to 2000 mg of a PRMT5 inhibitor having the formula (I) or the structure [ka] or a pharmaceutically acceptable salt thereof; [ka] (In the ceremony X 1 is NH, N(C1-C6 alkyl), O, or S; X 2 is N(C1-C6 alkyl), O, or S; Y 2 is H, C1-C6 alkyl, or C1-C6 haloalkyl; Z 1 and Z 2 each is independently H, F, or C1-C6 alkyl; and Z 3 , Z 4 , Z 5 , and Z 6 each of which is independently H, C1-C6 alkyl, or chloride; and (b) a second therapeutic agent selected from a PARP inhibitor, a KRAS inhibitor, a kinase-like protein 18A (KIF18A) inhibitor, or a kinase inhibitor; The method includes administering

[0006] In some embodiments, the PRMT5 inhibitor has the structure of Formula (S)-I, or a pharmaceutically acceptable salt thereof: [ka] It has.

[0007] In some embodiments, X 1 is O. In some embodiments, Z 1 and Z 2 are each H. In some embodiments, X 2 is O. In some embodiments, Z 3 , Z 4 , Z 5 , and Z 6 Each of Y is H. 2 is C1-C6 haloalkyl. 2 is CF3.

[0008] In some cases, PRMT5 inhibitors may: Compound B: [ka] or a salt thereof.

[0009] In some embodiments, the PRMT5 inhibitor is compound A: [ka] or a salt thereof.

[0010] In some embodiments, the PRMT5 inhibitor is compound G: [ka] or a salt thereof.

[0011] Pharmaceutically acceptable salts of the compounds described herein include those derived from suitable inorganic and organic acids and bases.

[0012] Combination therapy In some embodiments, the method further comprises administering a standard of care therapy to the patient as a combination therapy. The term "combination therapy," as used herein, refers to the administration of two or more therapeutic agents for treating cancer (e.g., a PRMT5 inhibitor as described herein and a second therapeutic agent selected from a PARP inhibitor, a KRAS inhibitor, a kinase-like protein 18A (KIF18A) inhibitor, or a kinase inhibitor). Such administration includes co-administration of these therapeutic agents substantially simultaneously, such as in a single capsule with a fixed active ingredient ratio. Alternatively, such administration includes co-administration in multiple containers or in separate containers for each active ingredient (e.g., tablets, capsules, powders, and liquids). The powders and / or liquids may be reconstituted or diluted to the desired dose before administration. In addition, such administration also includes the use of various therapeutic agents in a sequential manner, either at about the same time or at different times.

[0013] In some embodiments, the second therapeutic agent is a PARP inhibitor.Contemplated PARP inhibitors include, but are not limited to, olaparib, niraparib, rucaparib, and talazoparib.In some cases, the PARP inhibitor is olaparib.Olaparib is indicated as monotherapy in patients with advanced ovarian cancer with adverse or suspected adverse germline BRCA mutations (as detected by an FDA-approved test) who have been treated with three or more lines of chemotherapy.The recommended dose of olaparib for this indication is 400 mg (eight 50 mg capsules) taken twice daily, for a total daily dose of 800 mg.In some embodiments, the methods described herein comprise administering 400 mg twice daily to patients.

[0014] In some embodiments, the second therapeutic agent is a KRAS inhibitor. Contemplated KRAS inhibitors include, but are not limited to, sotorasib (Amgen), adagrasib (MRTX849, Mirati Therapeutics), JDQ443 (Novartis Pharmaceuticals), GDC-6036 (Genentech), D-1553 (InventisBio), LY3537982 (Eli Lilly and Company), BI 1823911 (Boehringer Ingelheim), JAB-21822 (Jacobio Pharmaceuticals), MK-1084 (Merck), YL-15293 (Shanghai YingLi Pharmaceutical Co.), RMC-6291 (Revolution Medicines), HBI-2438 (HUYABIO International), D3S-001 (D3 Bio (Wuxi) Co.), APG-1842 (Ascentage Pharma), VRTX126 (VRise Therapeutics), AZD4625 (AstraZeneca), ASP2453 (Astellas Pharma), ERAS-3490 (Erasca), JNJ-74699157 (ARS-3248, Janssen Research & Development), and BI1701963 (Boehringer Ingelheim). In some cases, the KRAS inhibitor is sotorasib. Sotorasib inhibits KRAS G12C Sotorasib is a small molecule that irreversibly inhibits mutant proteins. Sotorasib, also known as AMG 510 or 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-4-[(2S)-2-methyl-4-(prop-2-enoyl)piperazin-1-yl]pyrido[2,3-d]pyrimidin-2(1H)-one, has the following structure: [ka] It has.

[0015] For dosage information, see LUMAKRAS® US Prescribing Information, Amgen Inc., Thousand Oaks, California, 91320 (revision 11 / 2022), which is incorporated herein by reference in its entirety. In some embodiments, the methods disclosed herein comprise administering to a patient 960 mg sotorasib once daily. In some embodiments, the methods disclosed herein comprise administering to a patient 240 mg sotorasib once daily.

[0016] In some embodiments, the second therapeutic agent is a KIF18A inhibitor. The term "KIF18A inhibitor" refers to any compound useful for modulating KIF18A protein alone or in a complex bound to microtubules (MTs) to treat KIF18A-mediated pathologies and / or diseases, such as cancer. In some embodiments, the KIF18A inhibitor is N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxyethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide, which has the following structure: [ka] It has.

[0017] In some embodiments, the second therapeutic agent is a kinase inhibitor. Kinase inhibitors contemplated include, but are not limited to, palbociclib, trametinib, bosutinib, crizotinib, dasatinib, erlotinib, osimertinib, gefitinib, lapatinib, pazopanib, ruxolitinib, sunitinib, and vemurafenib. In some embodiments, the kinase inhibitor is palbociclib. In some embodiments, the kinase inhibitor is trametinib.

[0018] Dosage regimen A "therapeutically effective amount" of a PRMT5 inhibitor refers to an amount effective to treat or prevent the onset of, or alleviate existing symptoms of, a patient under treatment. Determining an effective amount is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein. Generally, a "therapeutically effective dose" refers to an amount of a PRMT5 inhibitor described herein that results in the realization of a desired effect. For example, a therapeutically effective amount of a PRMT5 inhibitor described herein reduces MTAP activity by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90% compared to a control.

[0019] In a specific embodiment, the PRMT5 inhibitors described herein are orally administered once daily to a patient in need thereof. A "patient" or "subject" to which administration is contemplated includes, but is not limited to, a human (i.e., a male or female of any age group, e.g., a pediatric subject (e.g., an infant, a child, an adolescent) or an adult subject (e.g., a young adult, a middle-aged adult, or an elderly adult). The terms "human," "patient," and "subject" are used interchangeably herein.

[0020] In some embodiments, the method comprises administering a PRMT5 inhibitor described herein in an amount ranging from 40 mg to 2000 mg. Optionally, the PRMT5 inhibitor is administered in divided daily doses, such as 2, 3, 4, 5, or 6 times daily. In some embodiments, the method comprises administering 40 mg, 120 mg, 240 mg, 480 mg, 960 mg, 1600 mg, or 2000 mg of a PRMT5 inhibitor to a patient once daily.

[0021] cancer In some embodiments, the cancer is an MTAP-deficient cancer. MTAP-deficient (or "MTAP-null") cancer refers to a cancer that lacks expression of the enzyme methylthioadenosine phosphorylase (MTAP). The MTAP gene, located at chromosome locus 9p21, is frequently co-deleted with the CDKN2A and CDKN2B genes. Selective MTAP deficiency refers to a defect without co-deletion of the CDKN2 gene, due to either selective deletion of the MTAP locus or methylation of the MTAP promoter. In MTAP-null cancer, at least 1% of diseased cells contain an MTAP deficiency. The terms "MTAP-null" and "MTAP-deficient" are used interchangeably herein.

[0022] In some embodiments, the cancer is an MTAP-deficient cancer and / or an MTA-accumulating cancer. An "MTAP-deficiency-associated" or "MTAP-deficient" or "MTAP-deficient" disease (e.g., a proliferative disease, e.g., cancer) or an "MTAP-associated" disease (e.g., a proliferative disease, e.g., cancer) or a "characterized by MTAP deficiency" disease (e.g., a proliferative disease, e.g., cancer) or the like refers to a condition (e.g., a proliferative disease, e.g., cancer) in which a large number of cells are MTAP-deficient. For example, in an MTAP-deficiency-associated disease, one or more diseased cells may have significantly reduced post-translational modification, production, expression, levels, stability, and / or activity of MTAP. Examples of MTAP-deficiency-associated diseases include, but are not limited to, cancers including glioblastoma, malignant peripheral nerve sheath tumor (MPNST), esophageal cancer (e.g., esophageal squamous cell carcinoma or esophageal adenocarcinoma), bladder cancer (e.g., bladder urothelial carcinoma), pancreatic cancer (e.g., pancreatic adenocarcinoma), mesothelioma, melanoma, non-small cell lung cancer (NSCLC; e.g., lung squamous cell or lung adenocarcinoma), astrocytoma, undifferentiated pleomorphic sarcoma, diffuse large B-cell lymphoma (DLBCL), leukemia, head and neck cancer, gastric adenocarcinoma, myxofibrosarcoma, bile duct sarcoma, brain cancer, stomach cancer, kidney cancer, breast cancer, endometrial cancer, urinary tract cancer, liver cancer, soft tissue cancer, pleural cancer, and colorectal cancer or sarcoma. In patients with MTAP-deficiency-associated diseases, it is possible that some diseased cells (e.g., cancer cells) may be MTAP-deficient, while others are not. Similarly, some diseased cells may have MTA accumulation, while others do not. Thus, the present disclosure encompasses therapeutic methods involving diseases of these tissues, or any other tissues, in which the proliferation of MTAP-deficient cells and / or MTA-accumulating cells can be inhibited by administering a PRMT5 inhibitor. Some cancer cells that are MTAP-deficient are also CDKN2A-deficient; the post-translational modification, production, expression, level, stability, and / or activity of the CDKN2A gene or its product are reduced in these cells. The genes for MTAP and CDKN2A are located in close proximity on chromosome 9p21; MTAP is located approximately 100 kb telomeric to CDKN2A. Many cancer cell types harbor CDKN2A / MTAP loss (loss of both genes). Thus, in some embodiments, MTAP-deficient cells are also CDKN2A-deficient.

[0023] In some embodiments, the patient has a cancer that further comprises a KRAS G12C mutation. KRAS G12C mutations occur at the alteration frequencies shown in the table below (Cerami et al., Cancer Discov. 2012, 2(5), 401; Gao et al., Science Signaling 2013, 6(269), pl1). For example, this table shows that 11.6% of patients with non-small cell lung cancer have one or more cells with KRAS G12C It shows that cancers that express the protein are found.

[0024] [Table 1]

[0025] In some embodiments, the cancer is acute myeloid leukemia, adolescent cancer, childhood adrenocortical carcinoma, AIDS-related cancers (e.g., lymphoma and Kaposi's sarcoma), anal cancer, appendiceal cancer, astrocytoma, atypical teratoid, basal cell carcinoma, cholangiocarcinoma, bladder cancer, bone cancer, brain stem glioma, brain tumor, breast cancer, triple-negative breast cancer, bronchial tumor, Burkitt's lymphoma, carcinoid tumor, atypical teratoid, embryonal tumor, germ cell tumor, primary lymphoma, cervical cancer, pediatric cancer, chordoma, cardiac tumor, chronic lymphocytic leukemia (CLL), chronic myelocytic leukemia (CML), chronic myeloproliferative disorders (CMD), and / or myeloproliferative disorders (MMD). disorder), colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS), embryonal tumors, CNS cancer, endometrial cancer, ependymoma, esophageal cancer, nasal neuroblastoma, Ewing's sarcoma, extracranial germ cell tumors, extragonadal germ cell tumors, eye cancer, osteofibrous histiocytoma, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors (GIST), germ cell tumors, gestational trophoblastic tumors, hairy cell leukemia, head and neck cancer, cardiac cancer, liver cancer, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumors, pancreatic neuroendocrine tumors, kidney cancer, laryngeal cancer, lip and oral cancer, liver cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma, metastatic squamous cell carcinoma of the neck of unknown primary origin, Midline tracheal cancer, oral cancer, multiple endocrine neoplasia syndrome, multiple myeloma / plasma cell neoplasm, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, multiple myeloma, Merkel cell carcinoma, malignant mesothelioma, malignant fibrous histiocytoma and osteosarcoma of bone, nasal and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer (NSCLC), oral cavity cancer, lip and oral cavity cancer, oropharyngeal cancer, ovarian cancer, high-grade serous ovarian cancer, pancreatic cancer, papillomatosis, paraganglioma, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pleuropulmonary blastoma, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, transitional cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, stomach cancer cancers such as gastric cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, renal pelvis and ureteral transitional cell carcinoma, trophoblastic tumor, rare childhood cancers, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, or virus-induced cancer.In some cases, the cancer is pancreatic cancer; esophageal cancer; melanoma; lung cancer; mixed Mullerian carcinoma; ovarian cancer; or gallbladder cancer.

[0026] In some embodiments, the cancer is glioblastoma, malignant peripheral nerve sheath tumor (MPNST), esophageal cancer (e.g., esophageal squamous cell carcinoma or esophageal adenocarcinoma), bladder cancer (e.g., bladder urothelial carcinoma), pancreatic cancer (e.g., pancreatic adenocarcinoma), mesothelioma, melanoma, non-small cell lung cancer (NSCLC; e.g., lung squamous cell or lung adenocarcinoma), astrocytoma, undifferentiated pleomorphic sarcoma, diffuse large B-cell lymphoma (DLBCL), leukemia, head and neck cancer, gastric adenocarcinoma, myxofibrosarcoma, bile duct sarcoma, brain cancer, stomach cancer, kidney cancer, breast cancer, endometrial cancer, urinary tract cancer, liver cancer, soft tissue cancer, pleural cancer, and colorectal cancer or sarcoma.

[0027] In some embodiments, the MTAP-null cancer is lung cancer, biliary tract cancer, head and neck squamous cell carcinoma, pancreatic adenocarcinoma, gallbladder cancer, or mesothelioma.

[0028] In some embodiments, the cancer is a solid tumor. In some embodiments, the tumor is malignant.

[0029] Exemplary MTAP-null solid tumors include, but are not limited to, MTAP-null brain cancers (including, but not limited to, MTAP-null glioma, MTAP-null oligodendroglioma, MTAP-null glioblastoma multiforme, MTAP-null astrocytoma, MTAP-null medulloblastoma, MTAP-null ependymoma, and MTAP-null meningioma), MTAP-null head and neck cancers (including, but not limited to, MTAP-null salivary gland (parotid) tumors, MTAP-null head and neck squamous cell carcinoma, and MTAP-null thyroid carcinoma), MTAP-null breast cancer (invasive ductal carcinoma, mixed mucinous carcinoma of the breast, and lobular carcinoma), MTAP-null mesothelioma, MTAP-null gastrointestinal cancer (including but not limited to MTAP-null esophageal cancer (including but not limited to adenocarcinoma and squamous cell carcinoma), MTAP-null esophagogastric junction cancer, MTAP-null gastric cancer (including but not limited to adenocarcinoma and signet ring cell carcinoma), MTAP-null small intestinal cancer, MTAP-null colon cancer, MTAP-null rectal cancer, and MTAP-null gastrointestinal stromal tumor), MTAP-null neuroendocrine tumor, M TAP-null hepatobiliary cancer (including but not limited to MTAP-null biliary cancer (including cholangiocarcinoma, gallbladder cancer, and ampullary cancer) and MTAP-null hepatocellular carcinoma), MTAP-null pancreatic cancer (including pancreatic adenocarcinoma), MTAP-null renal cancer (including but not limited to MTAP-null renal cell carcinoma), MTAP-null adrenocortical carcinoma, MTAP-null bladder cancer (including but not limited to MTAP-null urothelial carcinoma), MTAP-null adrenocortical carcinoma, MTAP-null endometrial cancer, MTAP-null uterine cancer, MTAP-null testicular cancer, MTA These include P-null germ cell tumors, or MTAP-null prostate cancer, MTAP-null sarcoma or MTAP-null bone cancer (including but not limited to MTAP-null osteosarcoma, MTAP-null chondrosarcoma, MTAP-null soft tissue sarcoma, MTAP-null Ewing's sarcoma, MTAP-null liposarcoma, MTAP-null leiomyosarcoma, and MTAP-null myxofibrosarcoma), MTAP-null skin tumors (MTAP-null cutaneous squamous cell carcinoma and MTAP-null melanoma), MTAP-null nerve sheath tumor, and MTAP-null carcinoma of unknown primary (CUP).

[0030] In some embodiments, the MTAP-null cancer is a hematological tumor. Exemplary hematological tumors include, but are not limited to, MTAP-null leukemia (including but not limited to MTAP-null acute lymphocytic leukemia, MTAP-null acute myeloid leukemia), MTAP-null lymphoma (including but not limited to MTAP-null mantle cell lymphoma, MTAP-null follicular lymphoma, MTAP-null diffuse large B-cell lymphoma, and MTAP-null mycosis fungoides).

[0031] Pharmaceutical Formulations and Routes of Administration Pharmaceutical compositions containing the PRMT5 inhibitors described herein can be manufactured in a conventional manner, for example, by conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilizing processes. Proper formulation depends on the chosen route of administration.

[0032] Monitoring Treatment Efficacy The effectiveness of a given cancer treatment can be determined by a skilled clinician. However, a treatment is considered to be an "effective treatment" as this term is used herein, for example, if any one or all of the signs or symptoms of a tumor are beneficially changed, or other clinically recognized symptoms are improved by at least 10% or even remitted after treatment with a drug, for example, as described herein. Efficacy can also be measured by an individual's lack of deterioration (i.e., the progression of the disease is halted), as assessed by the length of hospitalization or the need for medical intervention. Methods for measuring these indicators are known to those skilled in the art and / or are described herein.

[0033] In some embodiments, the combination therapy described herein exhibits the benefit of combination.The term " combination benefit " refers to the combination therapy being observed to be more effective than the treatment of any one individual therapy alone.In some embodiments, the combination therapy described herein exhibits the benefit of combination compared with PRMT5 monotherapy.In some embodiments, the combination therapy described herein exhibits the benefit of combination compared with the monotherapy of the second therapeutic agent described herein.

[0034] The description of the embodiments of the present disclosure is not intended to be exhaustive or to limit the disclosure to the precise form disclosed. Specific embodiments of, and examples for, the present disclosure are described herein for illustrative purposes; however, as those skilled in the art will recognize, various equivalent modifications are possible within the scope of the present disclosure. The teachings of the disclosure provided herein can be applied to other procedures or methods, as appropriate. The various embodiments described herein may be combined to provide further embodiments. Aspects of the present disclosure may be modified, if necessary, to employ compositions, functions, and concepts of the above references and applications to provide still further embodiments of the present disclosure. These and other changes can be made to the present disclosure in light of the detailed description.

[0035] Specific elements of any of the foregoing embodiments can be combined with or substituted for elements of other embodiments. Furthermore, although advantages associated with particular embodiments of the present disclosure are described in the context of those embodiments, other embodiments may also exhibit such advantages, and not necessarily all embodiments must exhibit such advantages to fall within the scope of the present disclosure.

[0036] All patents and other publications identified are expressly incorporated herein by reference for the purpose of describing and disclosing, for example, the methodology described in such publications that might be used in connection with the present invention. These publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representation as to the contents of these documents are based on the information available to the applicants and do not constitute any admission as to the correctness of the dates or contents of these documents.

[0037] Embodiments: 1. A method of treating cancer in a patient in need thereof, comprising administering to the patient: (a) an amount ranging from 40 mg to 2000 mg of a PRMT5 inhibitor, the amount being represented by Formula 1 or having the structure [ka] or a pharmaceutically acceptable salt thereof; [ka] (In the ceremony X 1 is NH, N(C1-C6 alkyl), O, or S; X 2 is N(C1-C6 alkyl), O, or S; Y 2 is H, C1-C6 alkyl, or C1-C6 haloalkyl; Z 1 and Z 2 each is independently H, F, or C1-C6 alkyl; and Z 3 , Z 4 , Z 5 , and Z 6wherein each of is independently H, C1-C6 alkyl, or chloride; and (b) a second therapeutic agent selected from a PARP inhibitor, a KRAS inhibitor, a kinase-like protein 18A (KIF18A) inhibitor, or a kinase inhibitor.

[0038] 2. The PRMT5 inhibitor has the structure of Formula (S)-I, or a pharmaceutically acceptable salt thereof: [ka] 2. The method of embodiment 1, comprising:

[0039] 3.X 1 3. The method of embodiment 1 or 2, wherein

[0040] 4.Z 1 and Z 2 4. The method of any one of embodiments 1 to 3, wherein each is H.

[0041] 5.X 2 5. The method of any one of embodiments 1 to 4, wherein

[0042] 6.Z 3 , Z 4 , Z 5 , and Z 6 6. The method of any one of embodiments 1-5, wherein each of

[0043] 7.Y 2 The method of any one of embodiments 1 to 6, wherein is C1-C6 haloalkyl.

[0044] 8.Y 2 8. The method of embodiment 7, wherein is CF3.

[0045] 9. The method of any one of embodiments 1-8, wherein the second therapeutic agent is a PARP inhibitor.

[0046] 10. The method of embodiment 9, wherein the PARP inhibitor is olaparib, niraparib, rucaparib, or talazoparib.

[0047] 11. The method of any one of embodiments 1-10, wherein the PARP inhibitor is olaparib.

[0048] 12. To the patient: 12. The method of any one of embodiments 1-11, comprising administering: (a) 40 to 2000 mg of a PRMT5 inhibitor; and (b) 300 mg olaparib twice daily.

[0049] 13. The method of any one of embodiments 1-8, wherein the second therapeutic agent is a KRAS inhibitor.

[0050] 14. The method of embodiment 13, wherein the KRAS inhibitor is sotorasib, adagrasib, JNJ-74699157, LY3537982, BI1823911, BI1701963, GDC-6036, tetrahydroquinazoline, JAB-2122, ARS-3248, AZD4625, or MRTX1133.

[0051] 15. The method of any one of embodiments 1-8, 13 and 14, wherein the KRAS inhibitor is sotorasib.

[0052] 16. The method of embodiment 15, comprising administering to the subject: (a) 40 to 2000 mg of a PRMT5 inhibitor; and (b) 960 mg sotorasib once daily.

[0053] 17. The method of embodiment 15, comprising administering to the subject: (a) 40 to 2000 mg of a PRMT5 inhibitor; and (b) 240 mg sotorasib once daily.

[0054] 18. The method of any one of embodiments 1-8, wherein the second therapeutic agent is a kinase-like protein 18A inhibitor.

[0055] 19. The method of any one of embodiments 1-8, wherein the second therapeutic agent is a kinase inhibitor.

[0056] 20. The method of embodiment 19, wherein the kinase inhibitor is palbociclib or trametinib.

[0057] 21. The method of embodiment 20, wherein the kinase inhibitor is palbociclib.

[0058] 22. The method of embodiment 21, comprising administering to the subject: (a) 40 to 2000 mg of a PRMT5 inhibitor; and (b) 125 mg palbociclib once daily.

[0059] 23. The method of embodiment 20, wherein the kinase inhibitor is trametinib.

[0060] 24. The method of embodiment 23, comprising administering to the subject: (a) 40 to 2000 mg of a PRMT5 inhibitor; and (b) 2 mg of trametinib once daily.

[0061] 25.PRMT5 inhibitors, [ka] 25. The method of any one of embodiments 1 to 24, wherein the compound has the structure:

[0062] 25.PRMT5 inhibitors, [ka] 25. The method of any one of embodiments 1 to 24, wherein the compound has the structure:

[0063] 26.PRMT5 inhibitor is Compound A: [ka] 25. The method of any one of embodiments 1 to 24, wherein the compound has the structure:

[0064] 27. The method according to any one of embodiments 1-26, wherein the PRMT5 inhibitor and the PARP inhibitor are administered simultaneously.

[0065] 28. The method of embodiment 11, wherein the PRMT5 inhibitor and olaparib are administered simultaneously.

[0066] 29. The method according to any one of embodiments 1-26, wherein the PRMT5 inhibitor and the PARP inhibitor are administered sequentially.

[0067] 30. The method of embodiment 11, wherein the PRMT5 inhibitor and olaparib are administered sequentially.

[0068] 31. The method according to any one of embodiments 1-26, wherein the PRMT5 inhibitor and the KRAS inhibitor are administered simultaneously.

[0069] 32. The method of embodiment 15, wherein the PRMT5 inhibitor and sotorasib are administered simultaneously.

[0070] 33. The method according to any one of embodiments 1-26, wherein the PRMT5 inhibitor and the KRAS inhibitor are administered simultaneously.

[0071] 34. The method of embodiment 15, wherein the PRMT5 inhibitor and sotorasib are administered sequentially.

[0072] 35. The method of embodiment 18, wherein the PRMT5 inhibitor and the KIF18A inhibitor are administered simultaneously.

[0073] 36. The method of embodiment 18, wherein the PRMT5 inhibitor and the KIF18A inhibitor are administered sequentially.

[0074] 37. The method according to any one of embodiments 1-26, wherein the PRMT5 inhibitor and the kinase inhibitor are administered sequentially.

[0075] 38. The method of embodiment 21, wherein the PRMT5 inhibitor and palbociclib are administered sequentially.

[0076] 39. The method of embodiment 23, wherein the PRMT5 inhibitor and trametinib are administered sequentially.

[0077] 40. The method according to any one of embodiments 1-26, wherein the PRMT5 inhibitor and the kinase inhibitor are administered simultaneously.

[0078] 41. The method of embodiment 21, wherein the PRMT5 inhibitor and palbociclib are administered simultaneously.

[0079] 42. The method of embodiment 23, wherein the PRMT5 inhibitor and trametinib are administered simultaneously. [Example]

[0080] Example 1 - Combination of PRMT5 inhibitors with olaparib in breast cancer cell lines Breast cancer cell lines (SUM149PT and HCC1395) were treated with a combination of PRMT5 inhibitors (i.e., Compound B and Compound G) and olaparib for 6 days. PRMT5 inhibitors (e.g., Compound B and Compound G) were run at a 1.9-fold dilution series, and the combination partners were run at a 1.2- to 1.7-fold dilution series to create an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fraction affected (Fa) using the following formula:

number

[0081] Synergy analysis was performed using CalcuSyn software to determine CI scores based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 1 to 3. *CI values ​​(Calcusyn): strong synergy: 0.1-0.3; synergy: 0.3-0.7; moderate synergy: 0.7-0.85; slight synergy: 0.85-0.9; nearly additive: 0.9-1.1.

[0082] [Table 2]

[0083] [Table 3]

[0084] [Table 4]

[0085] Example 2 - Combination of PRMT5 inhibitors with sotorasib in pancreatic cancer cell lines Pancreatic cancer cell lines (MIAPACA2T2) were treated with a combination of PRMT5 inhibitors (i.e., Compound B or Compound G) and sotorasib for 6 days. The PRMT5 inhibitors (e.g., Compound B or Compound G) were run at a 1.9-fold dilution series, and the combination partners were run at 1.2-1.7-fold dilutions to create an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fraction affected (Fa) using the following formula:

number

[0086] Synergy analysis was performed using CalcuSyn software to determine CI scores based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 4 and 5 below. *CI values ​​(Calcusyn): strong synergy: 0.1-0.3; synergy: 0.3-0.7; moderate synergy: 0.7-0.85; slight synergy: 0.85-0.9; nearly additive: 0.9-1.1.

[0087] [Table 5]

[0088] [Table 6]

[0089] Example 3 - Combination of PRMT5 inhibitors with sotorasib in bladder cancer cell lines Bladder cancer cell line (UM-UC-3) was treated with a combination of a PRMT5 inhibitor (i.e., Compound B) and sotorasib for 6 days. The PRMT5 inhibitor (e.g., Compound B) was run at a 1.9-fold dilution series, and the combination partner was run at a 1.2- to 1.7-fold dilution series to create an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fraction affected (Fa) using the following formula:

number

[0090] Synergy analysis was performed using CalcuSyn software to determine CI scores based on the drug concentrations used and the corresponding Fa values. The results are shown in Table 6 below. *CI values ​​(Calcusyn): strong synergy: 0.1-0.3; synergy: 0.3-0.7; moderate synergy: 0.7-0.85; slight synergy: 0.85-0.9; nearly additive: 0.9-1.1.

[0091] [Table 7]

[0092] Example 4 - Combination of PRMT5 inhibitors with sotorasib in lung cancer cell lines Lung cancer cell lines (LU99) were treated with a combination of PRMT5 inhibitors (i.e., Compound B or Compound G) and sotorasib for 6 days. The PRMT5 inhibitors (i.e., Compound B or Compound G) were run at a 1.9-fold dilution series, and the combination partners were run at 1.2-1.7-fold dilutions to create an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fraction affected (Fa) using the following formula:

number

[0093] Synergy analysis was performed using CalcuSyn software to determine CI scores based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 7 and 8 below. *CI values ​​(Calcusyn): strong synergy: 0.1-0.3; synergy: 0.3-0.7; moderate synergy: 0.7-0.85; slight synergy: 0.85-0.9; nearly additive: 0.9-1.1.

[0094] [Table 8]

[0095] [Table 9]

[0096] Example 5 - Combination of PRMT5 inhibitors with kinase-like protein 18A (KIF18A) in pancreatic cancer cell lines Pancreatic cancer cell lines (PSN1 and MIAPACA2T2) were treated with a combination of PRMT5 inhibitors (i.e., Compound B or Compound G) and KIF18A for 6 days. The PRMT5 inhibitors (e.g., Compound B or Compound G) were run at a 1.9-fold dilution series, and the combination partners were run at 1.2-1.7-fold dilutions to create an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fraction affected (Fa) using the following formula:

number

[0097] Synergy analysis was performed using CalcuSyn software to determine CI scores based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 9 to 12 below. *CI values ​​(Calcusyn): strong synergy: 0.1-0.3; synergy: 0.3-0.7; moderate synergy: 0.7-0.85; slight synergy: 0.85-0.9; nearly additive: 0.9-1.1.

[0098] [Table 10]

[0099] [Table 11]

[0100] [Table 12]

[0101] [Table 13]

[0102] Example 6 - Combination of PRMT5 inhibitors with kinase-like protein 18A (KIF18A) in lung cancer cell lines Lung cancer cell lines (LU99) were treated with a combination of a PRMT5 inhibitor (e.g., Compound B) and KIF18A for 6 days. The PRMT5 inhibitor (i.e., Compound B) was diluted 1.9-fold, and the combination partner was diluted 1.2-1.7-fold to create an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fraction affected (Fa) using the following formula:

number

[0103] Synergy analysis was performed using CalcuSyn software to determine CI scores based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 13 to 15 below. *CI values ​​(Calcusyn): strong synergy: 0.1-0.3; synergy: 0.3-0.7; moderate synergy: 0.7-0.85; slight synergy: 0.85-0.9; nearly additive: 0.9-1.1.

[0104] [Table 14]

[0105] Example 7 - Combination of PRMT5 inhibitors with KRAS G12X inhibitors in lung cancer cell lines Lung cancer cell lines (A549) were treated for 6 days with a combination of PRMT5 inhibitors (e.g., Compound B and Compound G) and KRAS G12X inhibitors. The PRMT5 inhibitors (e.g., Compound B or Compound G) were run at a 1.9-fold dilution series, and the combination partners were run at 1.2- to 1.7-fold dilutions to create an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fraction affected (Fa) using the following formula:

number

[0106] Synergy analysis was performed using CalcuSyn software to determine CI scores based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 13 to 15 below. *CI values ​​(Calcusyn): strong synergy: 0.1-0.3; synergy: 0.3-0.7; moderate synergy: 0.7-0.85; slight synergy: 0.85-0.9; nearly additive: 0.9-1.1.

[0107] [Table 15]

[0108] [Table 16]

[0109] Example 8 - Combination of PRMT5 inhibitors with palbociclib in lung cancer cell lines Lung cancer cell lines (H292 and A549) were treated with a combination of PRMT5 inhibitors (e.g., Compound B and Compound G) and palbociclib for 6 days. The PRMT5 inhibitors (e.g., Compound B or Compound G) were run at a 1.9-fold dilution series, and the combination partners were run at 1.2- to 1.7-fold dilutions to create an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fraction affected (Fa) using the following formula:

number

[0110] Synergy analysis was performed using CalcuSyn software to determine CI scores based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 16 to 19 below. *CI values ​​(Calcusyn): strong synergy: 0.1-0.3; synergy: 0.3-0.7; moderate synergy: 0.7-0.85; slight synergy: 0.85-0.9; nearly additive: 0.9-1.1.

[0111] [Table 17]

[0112] [Table 18]

[0113] [Table 19]

[0114] [Table 20]

[0115] Example 9 - Combination of PRMT5 inhibitors with trametinib in lung cancer cell lines Lung cancer cell lines (A549) were treated with a combination of PRMT5 inhibitors (e.g., Compound B and Compound G) and trametinib for 6 days. The PRMT5 inhibitors (e.g., Compound B or Compound G) were run at a 1.9-fold dilution series, and the combination partners were run at 1.2- to 1.7-fold dilutions to create an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fraction affected (Fa) using the following formula:

number

[0116] Synergy analysis was performed using CalcuSyn software to determine CI scores based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 20 and 21 below. *CI values ​​(Calcusyn): strong synergy: 0.1-0.3; synergy: 0.3-0.7; moderate synergy: 0.7-0.85; slight synergy: 0.85-0.9; nearly additive: 0.9-1.1.

[0117] [Table 21]

[0118] [Table 22]

[0119] Example 10 - Combination of PRMT5 inhibitors with trametinib in pancreatic cancer cell lines Pancreatic cancer cell lines (MIAPACA2T2) were treated with a combination of PRMT5 inhibitors (i.e., Compound B and Compound G) and trametinib for 6 days. The PRMT5 inhibitors (e.g., Compound B or Compound G) were run at a 1.9-fold dilution series, and the combination partners were run at 1.2-1.7-fold dilutions to create an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fraction affected (Fa) using the following formula:

number

[0120] Synergy analysis was performed using CalcuSyn software to determine CI scores based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 22 and 23 below. *CI values ​​(Calcusyn): strong synergy: 0.1-0.3; synergy: 0.3-0.7; moderate synergy: 0.7-0.85; slight synergy: 0.85-0.9; nearly additive: 0.9-1.1.

[0121] [Table 23]

[0122] [Table 24]

[0123] Example 11 - Combination of PRMT5 inhibitors and sotorasib inhibited tumor growth in LU99 NSCLC xenografts Ten female NOD / SCID mice were implanted with LU99 NSCLC xenografts. The mean tumor volume in each group was 100–200 mm. 3Mice were assigned to two different study groups based on tumor volume and initiated on once-daily oral administration of vehicle or compound G (100 mg / kg) in combination with sotorasib (100 mg / kg). Plotted data represent TGI (tumor growth inhibition) for each group, n=10. The results show that the combination of compound G and sotorasib resulted in significant antitumor activity in LU99 NSCLC xenografts compared to either agent alone (see Figure 1).

[0124] Example 12 - Combination of PRMT5 inhibitors and sotorasib inhibited tumor growth in LU99 NSCLC xenografts Ten female NOD / SCID mice were implanted with LU99 NSCLC xenografts. The mean tumor volume in each group was 100–200 mm. 3 Mice were assigned to two different study groups based on tumor volume and initiated with once-daily oral administration of vehicle or Compound B (100 mg / kg) in combination with sotorasib (100 mg / kg). Plotted data represent TGI (tumor growth inhibition) for each group, n=10. The results show that the combination of Compound B and sotorasib resulted in significant antitumor activity in LU99 NSCLC xenografts compared to either agent alone (see Figure 2).

[0125] Example 13 - Combination of PRMT5 inhibitors with sotorasib inhibited tumor growth in LU5268 NSCLC xenografts Ten female NOD / SCID mice were implanted with LU5268 NSCLC xenografts. The mean tumor volume in each group was 100–200 mm. 3 Mice were assigned to two different study groups based on tumor volume and initiated with once-daily oral administration of vehicle or Compound B (100 mg / kg) in combination with sotorasib (100 mg / kg). Plotted data represent TGI (tumor growth inhibition) for each group, n=10. The results show that the combination of Compound B and sotorasib resulted in significant antitumor activity in LU5268 NSCLC xenografts compared to either agent alone (see Figure 3).

[0126] Example 14 - Combination of PRMT5 inhibitors and sotorasib inhibited cell viability in pancreatic cancer cell lines Pancreatic cancer cell lines (MIAPACA2) were treated with a combination of a PRMT5 inhibitor (e.g., Compound B) and sotorasib for 6 days. The PRMT5 inhibitor (e.g., Compound B) was run in a 3-fold dilution series, and the combination partner was run in a 2-fold dilution series to create a 6 x 10 dose matrix, including a DMSO-only control. After 6 days, cell viability was measured using a CellTiter-Glo luminescence assay. As shown in Figure 4, the combination of Compound B and sotorasib resulted in a decrease in MIAPACA2 cell viability. To determine cell growth after combination treatment, nuclei were counted over 8 days using an IncuCyte live cell imager. MIAPACA2 cells were treated with DMSO, 150 nM Compound B, 50 nM sotorasib, or the combination (150 nM Compound B + 50 nM sotorasib). The results are shown in Figure 5.

[0127] Example 15 - Combination of PRMT5 inhibitors with sotorasib in pancreatic cancer cell lines Pancreatic cancer cell line (MIAPACA2) was treated with a combination of a PRMT5 inhibitor (i.e., Compound B) and sotorasib for 6 days. The PRMT5 inhibitor (i.e., Compound B) was run at a 1.9-fold dilution series, and the combination partner was run at a 1.2- to 1.7-fold dilution series to create an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fraction affected (Fa) using the following formula:

number

[0128] Synergy analysis was performed using CalcuSyn software to determine CI scores based on the drug concentrations used and the corresponding Fa values. The results are shown in Table 24 below. *CI values ​​(Calcusyn): CI<1 indicates synergy; CI=1 indicates additivity; CI>1 indicates antagonism.

[0129] [Table 25]

[0130] Example 16 - Combination of PRMT5 inhibitors with sotorasib inhibited tumor growth in KRAS G12C mutant NSCLC and PDAC xenografts Ten female NOD / SCID mice were implanted with LU99 NSCLC, LU5268 NSCLC, or MisPaCa2 PDAC xenografts. The mean tumor volume in each group was 100–200 mm. 3 Mice were assigned to four different study groups based on tumor volume and initiated once-daily oral administration of vehicle, compound G (100 mg / kg), sotorasib (100 mg / kg), or the combination of compound G (100 mg / kg) and sotorasib (100 mg / kg). Plotted data represent TGI (tumor growth inhibition) for each group, n=10. Results show that the combination of compound G and sotorasib significantly inhibits tumor growth in MTAP-null, KRAS G12C-mutant NSCLC and PDAC xenografts (see Figures 6A-C).

[0131] Example 17 - Combination of PRMT5 inhibitors with osimeritinib in lung cancer cell lines MTAP-null NSCLC cancer cell line (H1650) was treated with a combination of Compound G and osimertinib for 6 days. Compound G was run at a 1.9-fold dilution series, and the combination partners were run at 1.2-1.7-fold dilutions to create an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fraction affected (Fa) using the following formula:

number

[0132] Synergy analysis was performed using CalcuSyn software to determine CI scores based on the drug concentrations used and the corresponding Fa values. The results are shown in Table 25. *CI values ​​(Calcusyn): strong synergy: 0.1-0.3; synergy: 0.3-0.7; moderate synergy: 0.7-0.85; slight synergy: 0.85-0.9; nearly additive: 0.9-1.1.

[0133] [Table 26]

[0134] Example 18 - Combination of PRMT5 inhibitors with erlotinib in lung cancer cell lines MTAP-null NSCLC cancer cell line (H1650) was treated with a combination of Compound G and erlotinib for 6 days. Compound G was run at a 1.9-fold dilution series, and the combination partners were run at 1.2- to 1.7-fold dilutions to create an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fraction affected (Fa) using the following formula:

number

[0135] Synergy analysis was performed using CalcuSyn software to determine CI scores based on the drug concentrations used and the corresponding Fa values. The results are shown in Table 26. *CI values ​​(Calcusyn): strong synergy: 0.1-0.3; synergy: 0.3-0.7; moderate synergy: 0.7-0.85; slight synergy: 0.85-0.9; nearly additive: 0.9-1.1.

[0136] [Table 27]

Claims

1. 1. A method of treating cancer in a patient in need thereof, comprising administering to said patient: (a) a PRMT5 inhibitor in an amount ranging from 40 mg to 2000 mg, wherein the PRMT5 inhibitor is a compound of Formula I: 【Chemistry 1】 (In the ceremony X １ is NH, N(C １ ~C ６ alkyl), O, or S; X ２ is N(C １ ~C ６ alkyl), O, or S; Y ２ is H, C １ ~C ６ Alkyl, or C １ ~C ６ haloalkyl; Z １ and Z ２ each independently is H, F, or C １ ~C ６ is alkyl; and Z ３ , Z ４ , Z ５ , and Z ６ each independently represents H, C １ ~C ６ alkyl, or chloride) A compound shown in the formula: 【Chemistry 2】 or a pharmaceutically acceptable salt thereof; and (b) a second therapeutic agent selected from a PARP inhibitor, a KRAS inhibitor, a kinase-like protein 18A (KIF18A) inhibitor, or a kinase inhibitor. Administering

2. The PRMT5 inhibitor has the structure of formula (S)-I, or a pharmaceutically acceptable salt thereof: 【Transformation 3】 2. The method of claim 1, comprising:

3. X １ The method of claim 1 or 2, wherein is O.

4. Z １ and Z ２ The method of any one of claims 1 to 3, wherein each is H.

5. X ２ The method of any one of claims 1 to 4, wherein is O.

6. Z ３ , Z ４ , Z ５ , and Z ６ The method of any one of claims 1 to 5, wherein each of

7. Y ２ is C １ ~C ６ The method of any one of claims 1 to 6, wherein the alkyl is haloalkyl.

8. Y ２ is CF ３ The method of claim 7, wherein

9. The method of any one of claims 1 to 8, wherein the second therapeutic agent is a PARP inhibitor.

10. 10. The method of claim 9, wherein the PARP inhibitor is olaparib, niraparib, rucaparib, or talazoparib.

11. The method of any one of claims 1 to 10, wherein the PARP inhibitor is olaparib.

12. to the patient, (a) 40 to 2000 mg of a PRMT5 inhibitor; and (b) 300 mg olaparib twice daily The method of any one of claims 1 to 11, comprising administering

13. The method of any one of claims 1 to 8, wherein the second therapeutic agent is a KRAS inhibitor.

14. 14. The method of claim 13, wherein the KRAS inhibitor is sotorasib, adagrasib, JNJ-74699157, LY3537982, BI1823911, BI1701963, GDC-6036, JAB-2122, ARS-3248, AZD4625, or MRTX1133.

15. The method of any one of claims 1 to 8, 13 and 14, wherein the KRAS inhibitor is sotorasib.

16. To the subject, (a) 40 to 2000 mg of a PRMT5 inhibitor; and (b) 960 mg sotorasib once daily 16. The method of claim 15, comprising administering

17. To the subject, (a) 40 to 2000 mg of a PRMT5 inhibitor; and (b) 240 mg sotorasib once daily 16. The method of claim 15, comprising administering

18. The method of any one of claims 1 to 8, wherein the second therapeutic agent is a kinase-like protein 18A inhibitor.

19. 19. The method of claim 18, wherein the kinase-like protein 18A inhibitor is KIF18A.

20. The method of any one of claims 1 to 8, wherein the second therapeutic agent is a kinase inhibitor.

21. 21. The method of claim 20, wherein the kinase inhibitor is palbociclib or trametinib.

22. 22. The method of claim 21, wherein the kinase inhibitor is palbociclib.

23. To the subject, (a) 40 to 2000 mg of a PRMT5 inhibitor; and (b) 125 mg palbociclib once daily 23. The method of claim 22, comprising administering

24. 22. The method of claim 21, wherein the kinase inhibitor is trametinib.

25. To the subject, (a) 40 to 2000 mg of a PRMT5 inhibitor; and (b) 2 mg trametinib once daily 25. The method of claim 24, comprising administering

25. The PRMT5 inhibitor has the structure: 【Chemistry 4】 The method according to any one of claims 1 to 25, wherein the compound has the formula:

26. The PRMT5 inhibitor has the structure: 【Transformation 5】 The method according to any one of claims 1 to 25, wherein the compound has the formula:

27. The PRMT5 inhibitor has the structure of Compound A: 【Transformation 6】 The method according to any one of claims 1 to 25, wherein the compound has the formula:

28. The method of any one of claims 1 to 27, wherein the cancer is an MTAP-null cancer.

29. The method of any one of claims 1 to 27, wherein the cancer is an MTAP-deficient cancer, an MTA-accumulating cancer, or a combination thereof.

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

31. 31. The method of claim 30, wherein the tumor is malignant.

32. The method of any one of claims 1 to 31, wherein the cancer is lung cancer.

33. The method of any one of claims 1 to 31, wherein the cancer is pancreatic cancer.

34. 34. The method of claim 33, wherein the MTAP-null cancer is lung cancer.

35. 35. The method of claim 34, wherein the lung cancer is non-squamous cell lung cancer (NSCLC).

36. 29. The method of claim 28, wherein the MTAP-null cancer is biliary tract cancer.

37. 29. The method of claim 28, wherein the MTAP-null cancer is head and neck squamous cell carcinoma.

38. 29. The method of claim 28, wherein the MTAP-null cancer is pancreatic adenocarcinoma.

39. 29. The method of claim 28, wherein the MTAP-null cancer is gallbladder cancer.

40. 29. The method of claim 28, wherein the MTAP-null cancer is mesothelioma.

41. 29. The method of claim 28, wherein the cancer is not a primary brain tumor or lymphoma.

42. A therapeutically effective amount of (a) a PRMT5 inhibitor in an amount ranging from 40 mg to 2000 mg, wherein the PRMT5 inhibitor is a compound of Formula I: 【Transformation 7】 (In the ceremony X １ is NH, N(C １ ~C ６ alkyl), O, or S; X ２ is N(C １ ~C ６ alkyl), O, or S; Y ２ is H, C １ ~C ６ Alkyl, or C １ ~C ６ haloalkyl; Z １ and Z ２ each independently is H, F, or C １ ~C ６ is alkyl; and Z ３ , Z ４ , Z ５ , and Z ６ each independently represents H, C １ ~C ６ alkyl, or chloride) or the compound shown in the structure: 【Transformation 8】 or a pharmaceutically acceptable salt thereof; and (b) a second therapeutic agent selected from a PARP inhibitor, a KRAS inhibitor, a kinase-like protein 18A (KIF18A) inhibitor, or a kinase inhibitor. Use of.

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