Cancer treatment using mta-combined PRMT5

Abstract

Described herein are methods of treating cancer in a patient 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 Art

[0001] The epigenetic regulation of gene expression is the important biological determinant of protein production and cell differentiation, and plays an important pathogenic effect in many human diseases.Epigenetic regulation relates to the heritable modification of genetic material, and does not change its nucleotide sequence.Typically, by carrying out selective and reversible modification (for example, methylation) to DNA and protein (for example, histone), epigenetic regulation is mediated, and the conformational transition between the transcriptional active state and the inactive state of the modification control chromatin is carried out.These covalent modifications can be controlled by enzymes such as methyltransferases (for example, PRMT5), and wherein many enzymes are associated with the specific genetic changes that may cause human diseases.PRMT5 works in diseases (such as proliferative disorders, metabolic disorders and blood disorders).

[0002] Homozygous deletion of tumor suppressor genes is a key driver of cancer and often results in the collateral loss of passenger genes located in the genome near the tumor suppressor. Loss of these passenger genes can create therapeutically tractable vulnerabilities unique to tumor cells. Homozygous deletion of the chromosome 9p21 locus (carrying the well-known tumor suppressor CDKN2A (cyclin-dependent kinase inhibitor 2A)) occurs in 15% of all tumors and often includes 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 has a very similar structure to S-adenosylmethionine (SAM), a substrate methyl donor for the type II methyltransferase PRMT5. Elevated MTA levels driven by the loss of MTAP selectively compete with SAM for binding to PRMT5, which places the methyltransferase in a hypomorphic state and is susceptible to further PRMT5 inhibition. Multiple genome-scale shRNA dropout screens performed in large tumor cell line groups have identified a strong correlation between MTAP loss and cell line dependence on PRMT5, further highlighting the strength of this metabolic loophole. However, PRMT5 is a known essential gene for cells, and conditional PRMT5 knockout and siRNA knockdown studies have shown that a significant burden may be associated with suppressing PRMT5 in normal tissues (e.g., pancytopenia, infertility, skeletal muscle loss, cardiac hypertrophy, etc.). Therefore, novel strategies are needed to exploit this metabolic loophole and preferentially target PRMT5 in MTAP-invalid tumors while not hindering PRMT5 in normal tissues (MTAP WT). Targeting PRMT5 with small molecule inhibitors of MTA collaboration can preferentially target the MTA-bound state of PRMT5 enriched in MTAP-invalid tumor cells while providing an improved therapeutic index that exceeds normal cells (wherein MTAP is complete and MTA levels are low). Summary of the Invention

[0003] The present disclosure provides methods of treating cancer in a patient in need thereof, comprising administering to the patient:

[0004] (a) a PRMT5 inhibitor in an amount ranging from 40 mg to 2000 mg, wherein the PRMT5 inhibitor comprises a compound represented by formula (I) or having the structure A compound or a pharmaceutically acceptable salt thereof;

[0005]

[0006] in

[0007] X1 is NH, N(C1-C6 alkyl), O or S;

[0008] X 2 is N(C1-C6 alkyl), O, or S;

[0009] Y 2 is H, C1-C6 alkyl, or C1-C6 haloalkyl;

[0010] Z 1 and Z 2 Each of is independently H, F, or C1-C6 alkyl; and

[0011] Z 3 、Z 4 、Z 5 , and Z 6 Each of is independently H, C1-C6 alkyl, or chloride; and

[0012] (b) a second therapeutic agent selected from a PARP inhibitor, a KRAS inhibitor, a kinase-like protein 18A (KIF18A) inhibitor, or a kinase inhibitor. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 is a graph showing that the combination of Compound G and sotolacib produced significant anti-tumor activity in LU99 NSCLC xenografts relative to either single agent alone.

[0014] Figure 2 is a graph showing that the combination of Compound B and sotolacib produced significant anti-tumor activity in LU99 NSCLC xenografts relative to either single agent alone.

[0015] Figure 3 is a graph showing that the combination of Compound B and sotolacib produced significant anti-tumor activity relative to either single agent alone in LU5268 NSCLC patient-derived xenografts.

[0016] Figure 4 is a graph showing that the combination of Compound B and Sotolacib results in decreased viability of pancreatic cancer (MIAPACA2) cells.

[0017] Figure 5 is a graph showing that the combination of Compound B and Sotolacib significantly reduces pancreatic cancer (MIAPACA2) cell counts.

[0018] Figures 6A-6C The combination of Compound G and sotolacib was shown to significantly inhibit tumor growth in MTAP-null, KRAS G12C-mutant NSCLC and PDAC xenografts. Figure 6A : LU99 (NSCLC CDX) tumors were implanted into mice. Figure 6B :LU5268 (NSCLC PDX) tumor. Figure 6C MiaPaCa2 (PDAC CDX) tumors. Vehicle, Compound G, and sotolacib were administered as indicated. Data represent mean ± SD, n = 10 per group. STATS: P values determined by linear mixed-effects model with Tukey's full group comparison: combination versus either agent alone; ****p < 0.0001. DETAILED DESCRIPTION

[0019] The present disclosure provides methods of treating cancer in a patient in need thereof, comprising administering to the patient:

[0020] (a) a PRMT5 inhibitor in an amount ranging from 40 mg to 2000 mg, wherein the PRMT5 inhibitor comprises a compound represented by formula (I) or having the structure A compound or a pharmaceutically acceptable salt thereof;

[0021]

[0022] in

[0023] X 1 is NH, N(C1-C6 alkyl), O or S;

[0024] X 2 is N(C1-C6 alkyl), O, or S;

[0025] Y 2 is H, C1-C6 alkyl, or C1-C6 haloalkyl;

[0026] Z 1 and Z 2 Each of is independently H, F, or C1-C6 alkyl; and

[0027] Z 3 、Z 4 、Z 5 , and Z 6 Each of is independently H, C1-C6 alkyl, or chloride; and

[0028] (b) a second therapeutic agent selected from a PARP inhibitor, a KRAS inhibitor, a kinase-like protein 18A (KIF18A) inhibitor, or a kinase inhibitor.

[0029] In some embodiments, the PRMT5 inhibitor has a structure of Formula (S)-1 or a pharmaceutically acceptable salt thereof:

[0030]

[0031] In some embodiments, X 1 is 0. In some embodiments, Z 1 and Z 2 Each is H. In some embodiments, X 2 is O. In some embodiments, Z 3 、Z 4 、Z 5 , and Z 6 Each of is H. In some embodiments, Y 2 is C1-C6 haloalkyl. In some embodiments, Y 2 It's CF3.

[0032] In some instances, the PRMT5 inhibitor is a compound having the following structure:

[0033] Compound B: or a salt thereof.

[0034] In some embodiments, the PRMT5 inhibitor is a compound having the structure of Compound A: or a salt thereof.

[0035] In some embodiments, the PRMT5 inhibitor is a compound having the structure of Compound G: or a salt thereof.

[0036] Pharmaceutically acceptable salts of the compounds described herein include those derived from suitable inorganic and organic acids and bases.

[0037] Combination therapy

[0038] In some embodiments, the method further includes administering standard of care therapy to the patient as a combination therapy. As used herein, the term "combination therapy" refers to administering two or more therapeutic agents (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) to treat cancer. Such administration encompasses co-administration of these therapeutic agents in a substantially simultaneous manner, such as administration with a single capsule of an active ingredient having a fixed ratio. Alternatively, such administration is encompassed in multiple containers or co-administered in a separate container (e.g., tablets, capsules, powders and liquids) of each active ingredient. Powder and / or liquid can be reconstituted or diluted to the desired dose before administration. In addition, such administration also encompasses the use of each type of therapeutic agent in sequence at approximately the same time or at different times.

[0039] 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 a monotherapy for patients with advanced ovarian cancer who have deleterious or suspected deleterious germline BRCA mutations (as detected by an FDA-approved test) and who have been treated with three or more prior 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 to the patient twice daily.

[0040] In some embodiments, the second therapeutic agent is a KRAS inhibitor. Contemplated KRAS inhibitors include, but are not limited to, sotolacib (Amgen), adagracib (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., Ltd.), RMC-6291 (Revolution Pharmaceuticals), and LY3537982 (Eli Lilly and Company). medicines), HBI-2438 (HUYABIO International), D3S-001 (D3 Bio (Wuxi) Co., Ltd.), 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 instances, the KRAS inhibitor is sotolacib. Sotolacib is an irreversible inhibitor of KRAS G12C Small molecule that targets mutant proteins. Sotolacib is also known as AMG 510 or 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(prop-2-yl)pyridin-3-yl]-4-[(2S)-2-methyl-4-(prop-2-enoyl)piperazin-1-yl]pyrido[2,3-d]pyrimidin-2(1H)-one and has the following structure:

[0041]

[0042] Dosage information can be found in U.S. Prescribing Information, Amgen Inc., Thousand Oaks, California, 91320 (revised November 2022), which is incorporated herein by reference in its entirety. In some embodiments, the methods disclosed herein include administering 960 mg of sotolaxib to the patient once daily. In some embodiments, the methods disclosed herein include administering 240 mg of sotolaxib to the patient once daily.

[0043] In some embodiments, the second therapeutic agent is a KIF18A inhibitor. The term "KIF18A inhibitor" means any compound used to regulate KIF18A protein to treat KIF18A-mediated disorders and / or diseases (e.g., cancer) alone or in a binding complex with microtubules (MTs). 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 and has the following structure:

[0044]

[0045] In some embodiments, the second therapeutic agent is a kinase inhibitor. Contemplated kinase inhibitors include, but are not limited to, palbociclib, trametinib, bosutinib, crizotinib, dasatinib, erlotinib, osimertinib, gefitinib, lapatinib, pazoparib, ruxolitinib, sunitinib, and vemurafenib. In some embodiments, the kinase inhibitor is palbociclib. In some embodiments, the kinase inhibitor is trametinib.

[0046] Dosage regimen

[0047] A "therapeutically effective amount" of a PRMT5 inhibitor is an amount that effectively treats or prevents the development of existing symptoms in the subject being treated or alleviates existing symptoms. Determination of an effective amount is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein. Typically, a "therapeutically effective dose" refers to the amount of a PRMT5 inhibitor described herein that results in achieving the 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.

[0048] In a specific embodiment, the PRMT5 inhibitors described herein are orally administered once daily to patients in need thereof. "Patients" or "subjects" considered for administration include, but are not limited to, people (i.e., males or females of any age group, such as pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults or the elderly)). The terms "human," "patient," and "subject" are used interchangeably herein.

[0049] In some embodiments, these methods include administering a PRMT5 inhibitor as described herein in an amount ranging from 40 mg to 2000 mg. In some cases, the PRMT5 inhibitor is administered in a separate daily dose, such as twice a day, three times, four times, five times, or six times. In some embodiments, these methods include 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.

[0050] cancer

[0051] 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 chromosomal locus 9p21, is frequently co-deleted with the CDKN2A and CDKN2B genes. Selective MTAP deficiency refers to a deficiency in the absence of CDKN2 gene co-deletion due to selective deletion of the MTAP locus or methylation of the MTAP promoter. MTAP-null cancers include MTAP deficiency in at least 1% of diseased cells. The terms "MTAP-null" and "MTAP-deficient" are used interchangeably herein.

[0052] In some embodiments, the cancer is MTAP-deficient and / or MTA-accumulating cancer. "MTAP-deficiency-associated" or "MTAP-deficient" or "MTAP-deficient" disease (e.g., a proliferative disease, such as cancer) or a disease "associated with MTAP deficiency" (e.g., a proliferative disease, such as cancer) or a disease "characterized by MTAP deficiency" (e.g., a proliferative disease, such as cancer), etc., refers to a disease in which a large number of cells are MTAP-deficient cells (e.g., a proliferative disease, such as cancer). For example, in an MTAP-deficient-associated disease, one or more disease cells may have significantly reduced post-translational modification, production, expression, level, stability and / or activity of MTAP. Examples of MTAP-deficient associated diseases include, but are not limited to, cancers including, but not limited to, 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 carcinoma or lung adenocarcinoma), astrocytoma, undifferentiated pleomorphic sarcoma, diffuse large B-cell lymphoma (DLBCL), leukemia, head and neck cancer, gastric adenocarcinoma, myxofibrosarcoma, bile duct cancer, brain cancer, gastric 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-deficient associated diseases, some disease cells (e.g., cancer cells) may be MTAP-deficient cells, while other disease cells are not. Similarly, some disease cells may be MTA-accumulating cells, while other disease cells are not. Thus, the present disclosure encompasses methods for treating diseases involving these tissues, or any other tissues, in which the proliferation of MTAP-deficient and / or MTA-accumulating cells can be inhibited by administering a PRMT5 inhibitor. Some MTAP-deficient cancer cells also lack CDKN2A; post-translational modification, production, expression, levels, 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 from the telomere of CDKN2A. Many cancer cell types contain CDKN2A / MTAP loss (loss of both genes). Thus, in some embodiments, MTAP-deficient cells also lack CDKN2A.

[0053] In some embodiments, the patient has a cancer that further comprises a KRAS G12C mutation. The frequency of changes in the KRAS G12C mutation is shown in the following table (Cerami et al., Cancer Discov. 2012, 2(5), 401; Gao et al., Science Signaling 2013, 6(269), p11). For example, the table shows that 11.6% of patients with non-small cell lung cancer have a cancer in which one or more cells express KRAS G12C. G12C protein.

[0054] Table A

[0055]

[0056]

[0057] In some embodiments, the cancer is a cancer such as acute myeloid leukemia, juvenile cancer, childhood adrenocortical carcinoma, AIDS-related cancers (e.g., lymphoma and Kaposi's sarcoma), anal cancer, appendix cancer, astrocytoma, atypical teratoid, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, glioma, brain tumor, breast cancer, triple-negative breast cancer, bronchogenic carcinoma, Burkitt's lymphoma, carcinoid tumor, atypical teratoid, embryonal tumor, germ cell tumor, primary lymphoma, cervical cancer, childhood cancer, chordoma, heart tumor, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML) , chronic myeloproliferative disorder, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS), embryonal tumor, CNS cancer, endometrial cancer, ependymoma, esophageal cancer, nasal glioma, Ewing sarcoma, extracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, fibrous histiocytoma of bone, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic tumor, hairy cell leukemia, head and neck cancer, heart cancer, liver cancer, Hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumor, pancreatic neuroendocrine tumor, kidney cancer, Laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma, metastatic squamous neck cancer with occult primary, midline duct cancer, oral cancer, multiple endocrine neoplasia syndrome, multiple myeloma / plasma cell neoplasms, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasms, multiple myeloma, Merkel cell carcinoma, malignant mesothelioma, malignant fibrous histiocytoma and osteosarcoma of bone, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer (NSCLC), oral cancer, lip and oral cavity cancer, oropharyngeal cancer, ovarian cancer, high-grade serous ovarian cancer In some instances, the cancer is pancreatic 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 (stomach / gastric) cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, pharyngeal cancer, thymoma and thymic cancer, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, trophoblastoma, rare childhood cancers, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, or a virally induced cancer. In some instances, the cancer is pancreatic cancer; esophageal cancer; melanoma; lung cancer; mixed Müllerian cancer; ovarian cancer; or gallbladder cancer.

[0058] 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., cystourethral carcinoma), pancreatic cancer (e.g., pancreatic adenocarcinoma), mesothelioma, melanoma, non-small cell lung cancer (NSCLC, e.g., lung squamous cell carcinoma or lung adenocarcinoma), astrocytoma, undifferentiated pleomorphic sarcoma, diffuse large B-cell lymphoma (DLBCL), leukemia, head and neck cancer, gastric adenocarcinoma, myxofibrosarcoma, bile duct cancer, 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.

[0059] 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.

[0060] In some embodiments, the cancer is a solid tumor. In some embodiments, the tumor is malignant.

[0061] Exemplary MTAP-null solid tumors include, but are not limited to, MTAP-null brain cancers (including, but not limited to, MTAP-null gliomas, MTAP-null oligodendrogliomas, MTAP-null glioblastoma multiforme, MTAP-null astrocytomas, MTAP-null medulloblastomas, MTAP-null ependymomas, and MTAP-null meningiomas), MTAP-null head and neck cancers (including, but not limited to, MTAP-null salivary gland (parotid) tumors, MTAP-null head and neck squamous cell carcinomas, and MTAP-null thyroid cancers), MTAP-null breast cancers (including, but not limited to, infiltrating ductal breast cancer, mixed mucinous breast cancer, and lobular carcinoma), MTAP-null mesothelioma, MTAP-null gastrointestinal cancers (including, but not limited to, MTAP-null esophageal cancer (including, but not limited to, adenocarcinomas and squamous cell carcinomas), MTAP-null gastroesophageal junction cancer, MTAP-null gastric cancer (including, but not limited to, adenocarcinomas and signet ring cell carcinomas), MTAP-null small intestine cancer, MTAP-null colon cancer, MTAP-null rectal cancer, and MTAP-null gastrointestinal stromal tumors), MTAP-null neuroendocrine tumors, MTAP-null liver cancer, and MTAP-null esophageal cancer. Biliary cancer (including but not limited to MTAP-null biliary cancer (including bile duct cancer, 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, MTAP-null germ cell tumor 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 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 schwannoma and MTAP-null cancer of unknown primary (CUP).

[0062] In some embodiments, the MTAP-null cancer is a hematological tumor. Exemplary hematological tumors include, but are not limited to, MTAP-null leukemias (including but not limited to MTAP-null acute lymphoblastic leukemia, MTAP-null acute myeloid leukemia), MTAP-null lymphomas (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).

[0063] Pharmaceutical formulations and routes of administration

[0064] Pharmaceutical compositions containing the PRMT5 inhibitors described herein can be manufactured in a conventional manner, eg, by conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes. Proper formulation depends on the chosen route of administration.

[0065] Monitoring treatment efficacy

[0066] The efficacy of a given cancer treatment method can be determined by a skilled clinician. However, after treatment with an agent as described herein, if, for example, any one or all signs or symptoms of a tumor undergo a beneficial change, or other clinically acceptable symptoms are improved or even alleviated by, for example, at least 10%, then the treatment is considered to be "effective treatment" as used herein. Efficacy can also be measured by the absence of worsening (i.e., cessation of disease progression) of an individual assessed by hospitalization or the need for medical intervention. Methods for measuring these indicators are known to those skilled in the art and / or described herein.

[0067] In some embodiments, the combination therapies described herein show a combination benefit. The term "combination benefit" refers to the observed efficacy of the combination therapy being higher than the treatment of any one of the individual therapies used alone. In some embodiments, the combination therapies described herein show a combination benefit compared to PRMT5 monotherapy. In some embodiments, the combination therapies described herein show a combination benefit compared to monotherapy with a second therapeutic agent as described herein.

[0068] The description of the embodiments of the present disclosure is not intended to be exhaustive or intended to limit the present disclosure to the precise form disclosed. Although for illustrative purposes, specific embodiments and examples of the present disclosure are described herein, as will be appreciated by those skilled in the relevant art, various equivalent modifications can be made within the scope of the present disclosure. The teachings of the disclosure provided herein can be suitably applied to other programs or methods. Various embodiments described herein can be combined to provide other embodiments. If necessary, the aspects of the present disclosure can be modified to provide other embodiments of the present disclosure with the composition, function and concept of the above-mentioned references and applications. According to this detailed description, these and other changes can be made to the present disclosure.

[0069] The specific elements of any of the foregoing embodiments may be combined or replaced with elements in other embodiments. In addition, although advantages associated with certain embodiments of the present disclosure are described in the context of these embodiments, other embodiments may also exhibit such advantages, and not all embodiments must exhibit such advantages to fall within the scope of the present disclosure.

[0070] All patents and other publications identified are expressly incorporated herein by reference to describe and disclose, for example, methodologies described in such publications that may 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 reason of prior invention or for any other reason. All statements as to the dates or statements 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.

[0071] Example:

[0072] 1. A method for treating cancer in a patient in need thereof, comprising administering to the patient (a) a PRMT5 inhibitor in an amount ranging from 40 mg to 2000 mg, wherein the PRMT5 inhibitor comprises a compound represented by Formula 1 or having a structure A compound or a pharmaceutically acceptable salt thereof;

[0073] in

[0074] X 1 is NH, N(C1-C6 alkyl), O or S;

[0075] X 2 is N(C1-C6 alkyl), O, or S;

[0076] Y 2 is H, C1-C6 alkyl, or C1-C6 haloalkyl;

[0077] Z 1 and Z 2 Each of is independently H, F, or C1-C6 alkyl; and

[0078] Z 3 、Z 4 、Z 5 , and Z 6 wherein each of the alkyl groups 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.

[0079] 2. The method of Example 1, wherein the PRMT5 inhibitor has a structure of Formula (S)-I or a pharmaceutically acceptable salt thereof:

[0080]

[0081] 3. The method of embodiment 1 or embodiment 2, wherein X 1 It's O.

[0082] 4. The method of any one of embodiments 1-3, wherein Z 1 and Z 2 Each is H.

[0083] 5. The method of any one of embodiments 1-4, wherein X 2 It's O.

[0084] 6. The method of any one of embodiments 1-5, wherein Z 3 、Z 4 、Z 5 , and Z 6 Each of is H.

[0085] 7. The method of any one of embodiments 1-6, wherein Y 2 It is a C1-C6 haloalkyl group.

[0086] 8. The method of embodiment 7, wherein Y 2 It's CF3.

[0087] 9. The method of any one of embodiments 1-8, wherein the second therapeutic agent is a PARP inhibitor.

[0088] 10. The method of embodiment 9, wherein the PARP inhibitor is olaparib, niraparib, rucaparib, or talazoparib.

[0089] 11. The method of any one of embodiments 1-10, wherein the PARP inhibitor is olaparib.

[0090] 12. The method of any one of embodiments 1-11, comprising administering to the patient:

[0091] (a) 40-2000 mg of a PRMT5 inhibitor; and (b) 300 mg of olaparib twice daily.

[0092] 13. The method of any one of embodiments 1-8, wherein the second therapeutic agent is a KRAS inhibitor.

[0093] 14. The method of embodiment 13, wherein the KRAS inhibitor is sotolacib, adagracib, JNJ-74699157, LY3537982, BI1823911, BI1701963, GDC-6036, tetrahydroquinazoline, JAB-2122, ARS-3248, AZD4625, or MRTX1133.

[0094] 15. The method of any one of embodiments 1-8, 13, and 14, wherein the KRAS inhibitor is sotolacib.

[0095] 16. The method of embodiment 15, comprising administering to the subject (a) 40-2000 mg of the PRMT5 inhibitor; and (b) 960 mg of sotolacib once daily.

[0096] 17. The method of embodiment 15, comprising administering to the subject (a) 40-2000 mg of a PRMT5 inhibitor; and (b) 240 mg of sotolacib once daily.

[0097] 18. The method of any one of embodiments 1-8, wherein the second therapeutic agent is a kinase-like protein 18A inhibitor.

[0098] 19. The method of any one of embodiments 1-8, wherein the second therapeutic agent is a kinase inhibitor.

[0099] 20. The method of embodiment 19, wherein the kinase inhibitor is palbociclib or trametinib.

[0100] 21. The method of embodiment 20, wherein the kinase inhibitor is palbociclib.

[0101] 22. The method of embodiment 21, comprising administering to the subject (a) 40-2000 mg of the PRMT5 inhibitor; and (b) 125 mg of palbociclib once daily.

[0102] 23. The method of embodiment 20, wherein the kinase inhibitor is trametinib.

[0103] 24. The method of embodiment 23, comprising administering to the subject (a) 40-2000 mg of the PRMT5 inhibitor; and (b) 2 mg of trametinib once daily.

[0104] 25. The method of any one of embodiments 1-24, wherein the PRMT5 inhibitor is A compound having a structure of or a salt thereof.

[0105] 25. The method of any one of embodiments 1-24, wherein the PRMT5 inhibitor is A compound having a structure of or a salt thereof.

[0106] 26. The method of any one of embodiments 1-24, wherein the PRMT5 inhibitor is a compound having the structure of Compound A: or a salt thereof.

[0107] 27. The method of any one of embodiments 1-26, wherein the PRMT5 inhibitor and the PARP inhibitor are administered simultaneously.

[0108] 28. The method of embodiment 11, wherein the PRMT5 inhibitor and Olaparib are administered simultaneously.

[0109] 29. The method of any one of embodiments 1-26, wherein the PRMT5 inhibitor and PARP inhibitor are administered sequentially.

[0110] 30. The method of embodiment 11, wherein the PRMT5 inhibitor and olaparib are administered sequentially.

[0111] 31. The method of any one of embodiments 1-26, wherein the PRMT5 inhibitor and the KRAS inhibitor are administered simultaneously.

[0112] 32. The method of embodiment 15, wherein the PRMT5 inhibitor and sotolacib are administered simultaneously.

[0113] 33. The method of any one of embodiments 1-26, wherein the PRMT5 inhibitor and the KRAS inhibitor are administered simultaneously.

[0114] 34. The method of embodiment 15, wherein the PRMT5 inhibitor and sotolacib are administered sequentially.

[0115] 35. The method of embodiment 18, wherein the PRMT5 inhibitor and the KIF18A inhibitor are administered simultaneously.

[0116] 36. The method of embodiment 18, wherein the PRMT5 inhibitor and the KIF18A inhibitor are administered sequentially.

[0117] 37. The method of any one of embodiments 1-26, wherein the PRMT5 inhibitor and the kinase inhibitor are administered sequentially.

[0118] 38. The method of embodiment 21, wherein the PRMT5 inhibitor and palbociclib are administered sequentially.

[0119] 39. The method of embodiment 23, wherein the PRMT5 inhibitor and trametinib are administered sequentially.

[0120] 40. The method of any one of embodiments 1-26, wherein the PRMT5 inhibitor and the kinase inhibitor are administered simultaneously.

[0121] 41. The method of embodiment 21, wherein the PRMT5 inhibitor and palbociclib are administered simultaneously.

[0122] 42. The method of embodiment 23, wherein the PRMT5 inhibitor and trametinib are administered simultaneously.

[0123] Examples

[0124] Example 1 - Combination of PRMT5 inhibitors and olaparib in breast cancer cell lines

[0125] Breast cancer cell lines (SUM149PT and HCC1395) were treated with a combination of PRMT5 inhibitors (ie, Compound B and Compound G) and Olaparib for 6 days. PRMT5 inhibitors (eg, Compound B and Compound G) were diluted in a 1.9-fold series, and the combination partners were diluted in a 1.2 to 1.7-fold series to produce a dose matrix of 8 x 10, including DMSO-only controls. Cell viability was measured by CellTiter-Glo luminescence assay. Raw luminescence values were converted to affected fractions (Fa) by the following equation:

[0126]

[0127] 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 1-3 below. *CI values (Calcusyn): Strong synergy: 0.1-0.3; Synergy: 0.3-0.7; Moderate synergy: 0.7-0.85; Mild synergy: 0.85-0.9; Near-additivity: 0.9-1.1.

[0128] Table 1. Representative Compound B and Olaparib concentrations, and corresponding combined Fa and CI scores in SUM149PT cells.

[0129]

[0130] Table 2. Representative Compound B and Olaparib concentrations, and corresponding combined Fa and CI scores in HCC1395 cells.

[0131]

[0132] Table 3. Representative Compound G and Olaparib concentrations, and corresponding combined Fa and CI scores in HCC1395 cells.

[0133]

[0134] Example 2 - Combination of PRMT5 inhibitor and sotolacib in pancreatic cancer cell lines

[0135] Pancreatic cancer cell line (MIAPACA2T2) was treated with a combination of PRMT5 inhibitor (ie, compound B or compound G) and sotolacib for 6 days. The PRMT5 inhibitor (eg, compound B or compound G) was diluted in a 1.9-fold series, and the combination partner was diluted in a 1.2 to 1.7-fold series to produce an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured by CellTiter-Glo luminescence assay. The raw luminescence value was converted to the affected fraction (Fa) by the following equation:

[0136]

[0137] Synergy analysis was performed using CalcuSyn software to determine the CI score 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; mild synergy: 0.85-0.9; near additivity: 0.9-1.1.

[0138] Table 4. Representative Compound B and sotolacib concentrations, and corresponding combined Fa and CI scores in MIAPACA2T2 cells.

[0139]

[0140]

[0141] Table 5. Representative Compound G and sotolacib concentrations, and corresponding combined Fa and CI scores in MIAPACA2T2 cells.

[0142]

[0143] Example 3 - Combination of a PRMT5 inhibitor and sotolacib in bladder cancer cell lines

[0144] Bladder cancer cell line (UM-UC-3) was treated with a combination of PRMT5 inhibitor (ie, compound B) and sotolacib for 6 days. The PRMT5 inhibitor (eg, compound B) was diluted in a 1.9-fold series, and the combination partner was diluted in a 1.2 to 1.7-fold series to generate an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured by CellTiter-Glo luminescence assay. Raw luminescence values were converted to affected fractions (Fa) by the following equation:

[0145]

[0146] Synergy analysis was performed using CalcuSyn software to determine the CI score 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; mild synergy: 0.85-0.9; near additivity: 0.9-1.1.

[0147] Table 6. Representative Compound B and sotolacib concentrations, and corresponding combined-3Fa and CI scores in UM-UC-3 cells.

[0148]

[0149] Example 4 - Combination of PRMT5 inhibitors and sotolacib in lung cancer cell lines

[0150] Lung cancer cell lines (LU99) were treated with a combination of a PRMT5 inhibitor (ie, Compound B or Compound G) and Sotolacib for 6 days. The PRMT5 inhibitor (ie, Compound B or Compound G) was diluted in a 1.9-fold series, and the combination partner was diluted in a 1.2- to 1.7-fold series to generate an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured by CellTiter-Glo luminescence assay. Raw luminescence values were converted to fractions affected (Fa) by the following equation:

[0151]

[0152] Synergy analysis was performed using CalcuSyn software to determine the CI score 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; mild synergy: 0.85-0.9; near additivity: 0.9-1.1.

[0153] Table 7. Representative Compound B and sotolacib concentrations, and corresponding combined Fa and CI scores in LU99 cells.

[0154]

[0155] Table 8. Representative Compound G and sotolacib concentrations, and corresponding combined Fa and CI scores in LU99 cells.

[0156]

[0157] Example 5 - PRMT5 inhibitors and kinase-like protein 18A in pancreatic cancer cell lines

[0158] Combination of (KIF18A)

[0159] Pancreatic cancer cell lines (PSN1 and MIAPACA2T2) were treated with a combination of a PRMT5 inhibitor (ie, Compound B or Compound G) and KIF18A for 6 days. The PRMT5 inhibitor (eg, Compound B or Compound G) was diluted in a 1.9-fold series, and the combination partner was diluted in a 1.2- to 1.7-fold series to generate an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured by CellTiter-Glo luminescence assay. Raw luminescence values were converted to fractions affected (Fa) by the following equation:

[0160]

[0161] 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 9-12 below. *CI values (Calcusyn): Strong synergy: 0.1-0.3; Synergy: 0.3-0.7; Moderate synergy: 0.7-0.85; Mild synergy: 0.85-0.9; Near-additivity: 0.9-1.1.

[0162] Table 9. Representative Compound B and KIF18A concentrations, and corresponding combined Fa and CI scores in PSN1 cells.

[0163]

[0164] Table 10. Representative Compound G and KIF18A concentrations, and corresponding combined Fa and CI scores in PSN1 cells.

[0165]

[0166]

[0167] Table 11. Representative Compound B and KIF18A concentrations, and corresponding combined Fa and CI scores in MIAPACA2T2 cells.

[0168]

[0169] Table 12. Representative Compound G and KIF18A concentrations, and corresponding combined Fa and CI scores in MIAPACA2T2 cells.

[0170]

[0171] Example 6 - PRMT5 inhibitors and kinase-like protein 18A in lung cancer cell lines

[0172] Combination of (KIF18A)

[0173] Lung cancer cell line (LU99) was treated with a combination of PRMT5 inhibitor (e.g., Compound B) and KIF18A for 6 days. The PRMT5 inhibitor (i.e., Compound B) was diluted in a 1.9-fold series, and the combination partner was diluted in a 1.2- to 1.7-fold series to generate an 8x10 dose matrix, including a DMSO-only control. Cell viability was measured by CellTiter-Glo luminescence assay. The raw luminescence value was converted to the affected fraction (Fa) by the following equation:

[0174]

[0175] Synergy analysis was performed using CalcuSyn software to determine the CI score based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 13-15 below. *CI values (Calcusyn): Strong synergy: 0.1-0.3; Synergy: 0.3-0.7; Moderate synergy: 0.7-0.85; Mild synergy: 0.85-0.9; Near-additivity: 0.9-1.1.

[0176] Table 13. Representative Compound B and KIF18A concentrations, and corresponding combined Fa and CI scores in LU99 cells.

[0177]

[0178] Example 7 - Combination of a PRMT5 inhibitor and a KRAS G12X inhibitor in lung cancer cell lines

[0179] Lung cancer cell line (A549) was treated with a combination of PRMT5 inhibitors (e.g., Compound B and Compound G) and KRAS G12X inhibitors for 6 days. The PRMT5 inhibitor (e.g., Compound B or Compound G) was diluted in a 1.9-fold series, and the combination partner was diluted in a 1.2 to 1.7-fold series to generate an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured by CellTiter-Glo luminescence assay. The raw luminescence value was converted to the affected fraction (Fa) by the following equation:

[0180]

[0181] Synergy analysis was performed using CalcuSyn software to determine the CI score based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 13-15 below. *CI values (Calcusyn): Strong synergy: 0.1-0.3; Synergy: 0.3-0.7; Moderate synergy: 0.7-0.85; Mild synergy: 0.85-0.9; Near-additivity: 0.9-1.1.

[0182] Table 14. Representative Compound B and KRAS G12X inhibitor concentrations, and corresponding combined Fa and CI scores in A549 cells.

[0183]

[0184] Table 15. Representative Compound G and KRAS G12X inhibitor concentrations, and corresponding combined Fa and CI scores in A549 cells.

[0185]

[0186]

[0187] Example 8 - Combination of PRMT5 inhibitors and palbociclib in lung cancer cell lines

[0188] 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 inhibitor (e.g., Compound B or Compound G) was diluted in a 1.9-fold series, and the combination partner was diluted in a 1.2 to 1.7-fold series to produce an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured by CellTiter-Glo luminescence assay. The raw luminescence value was converted to the affected fraction (Fa) by the following equation:

[0189]

[0190] Synergy analysis was performed using CalcuSyn software to determine the CI score based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 16-19 below. *CI values (Calcusyn): Strong synergy: 0.1-0.3; Synergy: 0.3-0.7; Moderate synergy: 0.7-0.85; Mild synergy: 0.85-0.9; Near-additivity: 0.9-1.1.

[0191] Table 16. Representative Compound B and palbociclib concentrations, and corresponding combined Fa and CI scores in H292 cells.

[0192]

[0193]

[0194] Table 17. Representative Compound G and palbociclib concentrations, and corresponding combined Fa and CI scores in H292 cells.

[0195]

[0196] Table 18. Representative Compound B and palbociclib concentrations, and corresponding combined Fa and CI scores in A549 cells.

[0197]

[0198] Table 19. Representative Compound G and palbociclib concentrations, and corresponding combined Fa and CI scores in A549 cells.

[0199]

[0200] Example 9 - Combination of PRMT5 inhibitors and trametinib in lung cancer cell lines

[0201] Lung cancer cell line (A549) was treated with a combination of PRMT5 inhibitors (ie, compound B and compound G) and trametinib for 6 days. PRMT5 inhibitors (eg, compound B or compound G) were diluted in a 1.9-fold series, and the combination partners were diluted in a 1.2- to 1.7-fold series to generate an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured by CellTiter-Glo luminescence assay. Raw luminescence values were converted to fractions affected (Fa) by the following equation:

[0202]

[0203] Synergy analysis was performed using CalcuSyn software to determine the CI score based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 20-21 below. *CI values (Calcusyn): Strong synergy: 0.1-0.3; Synergy: 0.3-0.7; Moderate synergy: 0.7-0.85; Mild synergy: 0.85-0.9; Near-additivity: 0.9-1.1.

[0204] Table 20. Representative Compound G and trametinib concentrations, and corresponding combined Fa and CI scores in A549 cells.

[0205]

[0206] Table 21. Representative Compound G and trametinib concentrations, and corresponding combined Fa and CI scores in A549 cells.

[0207]

[0208] Example 10 - Combination of PRMT5 inhibitors and trametinib in pancreatic cancer cell lines

[0209] Pancreatic cancer cell line (MIAPACA2T2) was treated with a combination of PRMT5 inhibitor (ie, compound B or compound G) and trametinib for 6 days. The PRMT5 inhibitor (eg, compound B or compound G) was diluted in a 1.9-fold series, and the combination partner was diluted in a 1.2 to 1.7-fold series to generate an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured by CellTiter-Glo luminescence assay. Raw luminescence values were converted to affected fractions (Fa) by the following equation:

[0210]

[0211] Synergy analysis was performed using CalcuSyn software to determine the CI score based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 22-23 below. *CI values (Calcusyn): Strong synergy: 0.1-0.3; Synergy: 0.3-0.7; Moderate synergy: 0.7-0.85; Mild synergy: 0.85-0.9; Near-additivity: 0.9-1.1.

[0212] Table 22. Representative Compound B and trametinib concentrations, and corresponding combined Fa and CI scores in MIAPACA2T2 cells.

[0213]

[0214] Table 23. Representative Compound G and trametinib concentrations, and corresponding combined Fa and CI scores in MIAPACA2T2 cells.

[0215]

[0216]

[0217] Example 11 - Combination of a PRMT5 inhibitor and sotolacib inhibits tumor growth in LU99 NSCLC xenografts

[0218] Ten female NOD / SCID mice were implanted with LU99 NSCLC tumor xenografts. The average tumor volume in each group was between 100 and 200 mm. 3Mice were divided into two different study groups based on tumor volume and started with vehicle or Compound G (100 mg / kg) (orally once daily) in combination with Sotolacib (100 mg / kg). The data plotted represent TGI (tumor growth inhibition), with n=10 per group. The results showed that the combination of Compound G and Sotolacib produced significant antitumor activity in LU99 NSCLC xenografts compared to either agent alone (see Figure 1 ).

[0219] Example 12 - Combination of a PRMT5 inhibitor and sotolacib inhibits tumor growth in LU99 NSCLC xenografts

[0220] Ten female NOD / SCID mice were implanted with LU99 NSCLC tumor xenografts. The average tumor volume in each group was between 100 and 200 mm. 3 Mice were divided into two different study groups based on tumor volume and started with vehicle or Compound B (100 mg / kg) (orally once daily) in combination with Sotolacib (100 mg / kg). The data plotted represent TGI (tumor growth inhibition), with n=10 per group. The results showed that the combination of Compound B and Sotolacib produced significant antitumor activity in LU99 NSCLC xenografts compared to either agent alone (see Figure 2 ).

[0221] Example 13 - Combination of a PRMT5 inhibitor and sotolacib inhibits LU5268 NSCLC

[0222] Tumor growth in xenografts

[0223] Ten female NOD / SCID mice were implanted with LU5268 NSCLC tumor xenografts. The average tumor volume in each group was between 100 and 200 mm. 3 Mice were divided into two different study groups based on tumor volume and started with vehicle or Compound B (100 mg / kg) (orally once daily) in combination with Sotolacib (100 mg / kg). The data plotted represent TGI (tumor growth inhibition), with n=10 per group. The results showed that the combination of Compound B and Sotolacib produced significant antitumor activity in LU5268 NSCLC xenografts compared to either agent alone (see Figure 3 ).

[0224] Example 14 - Combination of a PRMT5 inhibitor and sotolacib inhibits cell viability in pancreatic cancer cell lines

[0225] Pancreatic cancer cell line (MIAPACA2) was treated with a combination of PRMT5 inhibitor (e.g., Compound B) and Sotolacib for 6 days. The PRMT5 inhibitor (e.g., Compound B) was diluted in a 3-fold series, and the combination partner was diluted in a 2-fold series to generate a 6 x 10 dose matrix, including a DMSO only control. After 6 days, cell viability was measured by CellTiter-Glo luminescence assay. Figure 4 As shown, the combination of Compound B and Sotolacib resulted in decreased viability of MIAPACA2 cells. To assess cell growth after combined treatment, nuclear counts were performed on an IncuCyte live cell imager over 8 days. MIAPACA2 cells were treated with DMSO, 150 nM Compound B, 50 nM Sotolacib, or the combination (150 nM Compound B + 50 nM Sotolacib). The results were shown in Figure 5 Shown in.

[0226] Example 15 - Combination of a PRMT5 inhibitor and sotolacib in pancreatic cancer cell lines

[0227] Pancreatic cancer cell line (MIAPACA2) was treated with a combination of PRMT5 inhibitor (ie, compound B) and sotolacib for 6 days. The PRMT5 inhibitor (ie, compound B) was diluted in a 1.9-fold series, and the combination partner was diluted in a 1.2 to 1.7-fold series to generate an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured by CellTiter-Glo luminescence assay. The raw luminescence value was converted to the affected fraction (Fa) by the following equation:

[0228]

[0229] Synergy analysis was performed using CalcuSyn software to determine the CI score 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. C = 1 indicates additive effect. C > 1 indicates antagonism.

[0230] Table 24. Representative Compound B and sotolacib concentrations, and corresponding CI scores in MIAPACA2 cells.

[0231]

[0232]

[0233] Example 16 - Combination of a PRMT5 inhibitor and sotolacib inhibits tumor growth in KRAS G12C mutant NSCLC and PDAC xenografts

[0234] Ten female NOD / SCID mice were implanted with LU99 NSCLC, LU5268 NSCLC, or MisPaCa2 PDAC tumor xenografts. The average tumor volume in each group was between 100 and 200 mm. 3 Mice were divided into four different study groups according to tumor volume and started to be dosed with vehicle, compound G (100 mg / kg), sotolacib (100 mg / kg) (orally once daily) or a combination of compound G (100 mg / kg) and sotolacib (100 mg / kg). The data plotted represent TGI (tumor growth inhibition), with n=10 per group. The results showed that the combination of compound G and sotolacib significantly inhibited tumor growth in MTAP-null, KRAS G12C mutant NSCLC and PDAC xenografts (see Figures 6A-6C ).

[0235] Example 17 - Combination of PRMT5 inhibitors and osimertinib in lung cancer cell lines

[0236] MTAP-null NSCLC cancer cell line (H1650) was treated with a combination of Compound G and osimertinib for 6 days. Compound G was administered in a 1.9-fold dilution series, and the combination partners were administered in a 1.2- to 1.7-fold dilution series to generate 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 fractions affected (Fa) using the following equation:

[0237]

[0238] Synergy analysis was performed using CalcuSyn software to determine the CI score 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; mild synergy: 0.85-0.9; near additivity: 0.9-1.1.

[0239] Table 25. Representative Compound G and osimertinib concentrations, and corresponding combined Fa and CI scores in H1650 cells.

[0240]

[0241] Example 18 - Combination of PRMT5 inhibitors and erlotinib in lung cancer cell lines

[0242] MTAP-null NSCLC cancer cell line (H1650) was treated with a combination of compound G and erlotinib for 6 days. Compound G was diluted in a 1.9-fold series, and the combination partner was diluted in a 1.2- to 1.7-fold series to generate 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 fractions affected (Fa) using the following equation:

[0243]

[0244] Synergy analysis was performed using CalcuSyn software to determine the CI score 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; mild synergy: 0.85-0.9; near additivity: 0.9-1.1.

[0245] Table 26. Representative Compound G and erlotinib concentrations, and corresponding combined Fa and CI scores in H1650 cells.

[0246]

Claims

1. A method of treating cancer in a patient in need thereof, the method comprising administering to the patient: (a) a PRMT5 inhibitor in an amount ranging from 40 mg to 2000 mg, wherein the PRMT5 inhibitor comprises a compound represented by or having the structure A compound or a pharmaceutically acceptable salt thereof; in 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 of is independently H, F, or C1-C6 alkyl; and Z 3 , Z 4 , Z 5 , and Z 6 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.

2. The method of claim 1, wherein the PRMT5 inhibitor has a structure of Formula (S)-I, or a pharmaceutically acceptable salt thereof:

3. The method of claim 1 or claim 2, wherein X 1 It's O.

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

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

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

7. The method of any one of claims 1 to 6, wherein Y 2 It is a C1-C6 haloalkyl group.

8. The method of claim 7, wherein Y 2 It's CF3.

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

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

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

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

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

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

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

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

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

18. The method of any one of claims 1-8, wherein the second therapeutic agent is a kinase-like protein 18A inhibitor. The method of claim 18 , wherein the KIF18A inhibitor is KIF18A.

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

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

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

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

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

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

26. The method of any one of claims 1-25, wherein the PRMT5 inhibitor is or a salt thereof.

27. The method of any one of claims 1-25, wherein the PRMT5 inhibitor is A compound having a structure of or a salt thereof.

28. The method of any one of claims 1-25, wherein the PRMT5 inhibitor is a compound having the structure of Compound A: or a salt thereof.

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

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

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

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

33. The method of any one of claims 1-31, wherein the cancer is lung cancer.

34. The method of any one of claims 1-31, wherein the cancer is pancreatic cancer.

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

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

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

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

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

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

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

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

43. Use of a therapeutically effective amount of (a) a PRMT5 inhibitor in an amount ranging from 40 mg to 2000 mg and (b) a second therapeutic agent for treating cancer, wherein the PRMT5 inhibitor comprises a compound shown in <Formula I> or has the structure A compound or a pharmaceutically acceptable salt thereof; in 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 of is independently H, F, or C1-C6 alkyl; and Z 3 , Z 4 , Z 5 , and Z 6 Each of the alkyl groups is independently H, C1-C6 alkyl, or chloride; The second therapeutic agent is selected from a PARP inhibitor, a KRAS inhibitor, a kinase-like protein 18A (KIF18A) inhibitor, or a kinase inhibitor.

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