PRMT5 inhibitor for the treatment of cancer

Abstract

Provided herein are methods of treating cancer (e.g., a tumor in the central nervous system) using a protein arginine methyltransferase 5 (PRMT5) inhibitor or pharmaceutically acceptable salts or compositions thereof. Also provided herein are methods of treating cancer (e.g., a tumor in the central nervous system) using a PRMT5 inhibitor or pharmaceutically acceptable salts or compositions thereof in combination with a Cyclin-dependent kinase (CDK) 4 / 6 inhibitor.

Description

Attorney Docket No TGO-037WOPRMT5 INHIBITOR FOR THE TREATMENT OF CANCERFIELD OF DISCLOSURE

[0001] The disclosure relates generally to use of a compound for inhibiting protein arginine me thy I transferase 5 (PRMT5) in the treatment of cancer.BACKGROUND

[0002] Protein arginine methyltransferase 5 (PRMT5) is a ty pe II arginine methyltransferase that regulates essential cellular functions, including the regulation of cell cycle progression, apoptosis and the DNA-damage response (Koh, C. et al., Curr Mol Bio Ae 2015 l(I):19-28; Wu et al., Nat Rev Drug Discovery 2021 20(3)1200-216).Methylthioadenosine phosphorylase (MTAP) is a critical enzyme in the methionine salvage pathway, a six-step process that recycles methionine from the product of polyamine synthesis, methylthioadenosine (MTA). Loss of MTAP causes the accumulation of its substrate, MTA, which has been described to function as a S-adenosyl-l-methionine (SAM)-competitive PRMT5 inhibitor (Kryukov, G. V. et al.. Science, 2016351(6278): 1214-1218; Marjon, K. et al.. Cell Reports 2016: 15(3)1574-587). Data from genome-wide genetic perturbation screens using shRNA suggests a selective requirement for PRMT5 activity particularly in A / 7 / tP-deleted cancer cell lines (Kruykov et al., 2016; Marjon et al., 2016), It is proposed that the accumulation of MTA caused by E4P-deficiency (e.g, A£E4P-deletion) in these cell lines partially inhibits PRMT5, rendering those cells selectively sensitive to additional PRMT5 inhibition.

[0003] A PRMT5 inhibitor that leverages the accumulation of MTA by binding in an MTA-uncompetitive, non-competitive or mixed mode manner or in a MTA-cooperative binding manner may demonstrate selectivity' for MTAP-deficient (<?.g., MTAP-deleted) tumor cells. There are currently no PRMT5 therapies approved by the United States Food and Drug Administration that demonstrate selectivity' for MTAP-deficient (e.g., MTAP-deleted) cancer cell lines.

[0004] Accordingly, there is a need for PRMT5 inhibitors for treating diseases that have a deficiency (e.g., deletion) of the MTAP gene, such as cancers.1IPTS- 200237393 1Attorney Docket No TGO-037WOSUMMARY OF THE DISCLOSURE

[0005] The present disclosure provides use of compounds for inhibiting protein arginine methyltransferase 5 (PRMT5) in the treatment of cancer.

[0006] In one aspect, the present disclosure provides a method of treating a tumor in the central nervous system (CNS) in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a protein arginine methyltransferase 5 (PRMT5) inhibitor, wherein the PRMT5 inhibitor is a compound having the structural formul:F Compound 1;wherein the compound is administered as a free base or a pharmaceutically acceptable salt or a composition thereof. This disclosure is based, in part, on the discovery that Compound 1 is able to cross the brain- blood barrier and therefore is expected to reach the CNS after systemic administration.

[0007] In some embodiments, the tumor m the CNS is a glioma In some embodiments, the glioma is MTAP-deficient. In some embodiments, the glioma is MTA-accumulating. In some embodiments, the glioma is a glioblastoma.

[0008] In one aspect, the present disclosure provides a method of treating a cancer m a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a protein arginine methyltransferase 5 (PRMT5) inhibitor, wherein the PRMT5 inhibitor is a compound having the structural formula:F Compound I;Attorney Docket No TGO-037WOwherein the compound is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0009] In some embodiments, the cancer comprises a solid tumor with MTAP-deficiency or MTAP-deletion. In some embodiments, the cancer comprises glioma (e.g, glioblastoma), non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma.

[0010] In some embodiments of any of the foregoing aspects, the PRMT5 inhibitor is Compound 1 and is administered systemically. In some embodiments, the PRMT5 inhibitor is Compound 1 and is administered orally.

[0011] In some embodiments of any of the foregoing aspects, the PRMT5 inhibitor is administered in combination with a second therapeutic agent. In some embodiments, the PRMT5 inhibitor is administered in combination with a cyclin-dependent kinase (CDK) 4 / 6 inhibitor having the structural formula:Compound 2;wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof. This disclosure is based, in part, on the discover}' by the applicant of a synergistic effect between the PRMT5 inhibitor and Compound 2 in treating cancer (e.g, glioblastoma).[0012| In some embodiments, the method further comprises administering to the subject a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor having the structural formula:wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0013] In some embodiments, the subject also receives a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor having the structural formula:IPTS'200237393.1Attorney Docket No TGO-037WOCompoundwherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0014] In some embodiments, the CDK4 / 6 inhibitor is Compound 2 and is administered systemically. In some embodiments, the CDK4 / 6 inhibitor is Compound 2 and is administered orally.

[0015] In some embodiments, the present disclosure provides a method of treating a cancer m a subject in need thereof, the method compnsing administering to the subject a therapeutically effective amount of a protein arginine methyltransferase 5 (PRMT5) inhibitor and a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor. wherein the PRMT5 inhibitor is a compound having the structural formula:F Compound 1;wherein the CDK4 / 6 inhibitor is a compound having the structural formula:FCompound 2:wherein Compound 1 and Compound 2 are each administered as a free base or a pharmaceutically acceptable salt or a composition thereof

[0016] In an embodiment the cancer comprises a solid tumor with MTAP-deficiency. In an embodiment the cancer comprises glioma, non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma

[0017] In some embodiments, the present disclosure provides a method of treating a tumor in the CNS in a subject in need thereof, the method comprising administering to the subject a 4IPTS'200237393.1Attorney Docket No TGO-037WOtherapeutically effective amount of a protein arginine methyltransferase 5 (PRMT5) inhibitor and a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor, wherein the PRMT5 inhibitor is a compound having the structural formula:H2NNF—j—FF Compound 1;wherein the CDK4 / 6 inhibitor is a compound having the structural formula:FN' Compound 2;wherein Compound 1 and Compound 2 are each administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0018] In an embodiment the tumor in the CNS is a glioma. In an embodiment the glioma is a glioblastoma. In an embodiment the tumor in the CNS is M P-deficient. In an embodiment the tumor in the CNS is MTA-accumulating. In an embodiment the PRMT5 inhibitor is administered systemically. In an embodiment the PRMT5 inhibitor is administered orally. In an embodiment the CDK4 / 6 inhibitor is administered systemically. In an embodiment the CDK 4 / 6 inhibitor is administered orally.

[0019] In some embodiments, the present disclosure provides a protein arginine methyltransferase 5 (PRMT5) inhibitor for use in treating a cancer in a subject in need thereof, wherein the PRMT5 inhibitor is a compound having the structural formula:H2NN4FF Compound I;5IPTS'200237393.1Attorney Docket No TGO-037WOwherein the compound is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0020] In an embodiment the cancer comprises a solid tumor with MTAP-deficiency. In an embodiment the cancer comprises glioma, non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma. In an embodiment the PRMT5 inhibitor is administered systemically. In an embodiment the PRMT5 inhibitor is administered orally.

[0021] In an embodiment the subject also receives a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor having the structural formula:FXFCompound 2;wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof In an embodiment the CDK 4 / 6 inhibitor is administered systemically. In an embodiment the CDK 4 / 6 inhibitor is administered orally.

[0022] In some embodiments, the present disclosure provides a protein arginine methyl transferase 5 (PR. MT5) inhibitor for use in treating a tumor in the central nervous system (CNS) in a subject m need thereof, wherein the PRMT5 inhibitor is a compound having the structural formula:oF-1 F“FCompound I;wherein the compound is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.In an embodiment the tumor in the CNS is a glioma. In an embodiment the glioma is a glioblastoma. In an embodiment the tumor in the CNS is MTAP-deficient. In an embodiment the tumor in the CNS is MTA-accumulating.

[0023] In some embodiments, the present disclosure provides a protein arginine methyl transferase 5 (PRMT5) inhibitor for use in combination with a cyclin-dependent 6IPTS'200237393.1Attorney Docket No TGO-037WOkinase (CDK) 4 / 6 inhibitor in treating a cancer in a subject in need thereof, wherein: the PRMT5 inhibitor is a compound having the structural formula:F Compound 1;wherein Compound 1 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof; and the CDK4 / 6 inhibitor is a compound having the structural formula:Compound 2;wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof

[0024] In an embodiment the cancer comprises a solid tumor with MTAP-deficiency.

[0025] In an embodiment the cancer comprises glioma, non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma.

[0026] In some embodiments, the present disclosure provides a protein arginine methyl transferase 5 (PRMT5) inhibitor for use in combination with a cyclin-dependent kinase (CDK) 4 / 6 inhibitor in treating a tumor in the central nervous system (CNS) in a subject in need thereof, wherein: the PRMT5 inhibitor is a compound having the structural formula:IPTS'200237393.1Attorney Docket No TGO-037WOF Compound 1;wherein Compound 1 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof; andthe CDK4 / 6 inhibitor is a compound having the structural formula:wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.[0027J In an embodiment the tumor in the CNS is a glioma. In an embodiment the glioma is a glioblastoma. In an embodiment the tumor in the CNS is MTAP-deficient. In an embodiment the tumor in the CNS is MTA-accumulating. In an embodiment the PRMT5 inhibitor is administered systemically. In an embodiment the PRMT5 inhibitor is administered orally. In an embodiment the CDK4 / 6 inhibitor is administered systemically. In an embodiment the CDK 4 / 6 inhibitor is administered orally.BRIEF DESCRIPTION OF THE DRAWINGS

[0028] FIG, 1 is a waterfall plot for a multi-lineage cancer cell line panel profiled with Compound 1 in a 7-day CellTiter-Glo assay. The maximum effect (Amax) of Compound 1 at a concentration equal to 10X the IC50 of Compound I in the HAP1 MTAP-null cancer cell is reported. The cell lines are colored by MTAP status with blue indicating MTAP wild-type status and red indicating MTAP-null status.

[0029] FIGs. 2A-2D are graphs depicting antitumor activity of Compound 1 in MTAP-null patient derived xenograft models. FIG. 2A: squamous non-small cell lung cancer; Compound 1 was dosed at 30 mg / kg and 90 mg / kg twice daily (BID) for 65 days, then the animals were 8IPTS'200237393.1Attorney Docket No TGO-037WOmonitored for tumor regrowth: FIG. 2B: glioblastoma; Compound I was dosed at 30 mg / kg and 90 mg / kg twice daily (BID); for the 90 mg / kg group, the compound was dosed for 45 days, then the animals were monitored for tumor regrowth until day 66: FIG. 2C: pancreatic cancer: Compound 1 was dosed at 30 mg / kg and 90 mg / kg twice daily (BID) for the duration of the study; and FIG. 2D: cholangiocarcinoma; Compound 1 was dosed at 30 mg / kg and 90 mg / kg twice daily (BID) for the duration of the study. 5% DMA and 20% Captisol was used as vehicle in the study. Data are presented as mean ± SEM.

[0030] FIG. 3 is a graph depicting antitumor activity’ of Compound 1 in MTAP-null LU99 human non-small cell lung cancer xenograft model with Compond 1 dosed at ]0 mg / kg, 30 mg / kg, and 90 mg / kg twice daily’ (BID). 5% DMA and 20% Captisol was used as vehicle in the study. Data are presented as mean ± SEM.

[0031] FIG. 4A is a graph depicting the in vitro combination benefit of abemaciclib (Compound 2) and Compound 1 in LU99 MT P-deleted non-small cell lung cancer (NSCLC) cell line as measured through Cell-Titer Gio assay. FIG.4B is a graph depicting results from colony formation assay to test combination of Compound 1 (TNG456 in FIG.4B) and abemaciclib in LU99 MTAP-deleted non-small cell lung cancer (NSCLC) cell line.FIGs. 4C-4D are graphs depicting body weight change (%) (FIG. 4C) and antitumor activity (FIG. 4D) of Compound 1 (TNG456 in FIG. 4C-4D) in combination with abemaciclib in MTAP-null LU99 human non-small cell lung cancer xenograft model. Compound 1 and abemaciclib w-ere dosed at 30 mg / kg twice daily (BID) and 50 mg / kg once daily (QD), respectively. V ehicle for TNG456: 5% DMA+20% Captisol; vehicle for abemaciclib: 1% hydroxyethyl cellulose + 0 1 % antifoam in 25 mM PB pH 2; vehicle for combination was 5% DMA and 20% Captisol co-dosed with 1% hydroxy ethyl cellulose + 0.1% antifoam in 25 mM PB pH 2. Data are presented as mean ± SEM. ** refers to p-value = 0.0018; *** refers to p-value = 0.0004.

[0032] FIG. 5A is a line graph showing pharmacokinetics of Compound 1 (TNG456) in BALB / c mice following oral administration of Compound 1 (TNG456) at the indicated doses / schedule. after 7 days of dosing. FIG. 5A is also a bar graph show ing tumor pharmacodynamic analysis at indicated post last dose time point. Normalized (relative to vehicle) symmetric dimethylarginine (SDMA)-modified protein levels are depicted as mean + / - SEM. FIGs. 5B-5C are graphs depicting body weight change (%) (FIG. 5B) and antitumor activity’ (FIG. 5C) of Compound 1 (TNG456 in FIG. 5A-5B) in U87MG MTAP- null glioblastoma xenograft model with Compound 1 dosed at 10 mg / kg, 30 mg / kg, and 909IPTS- 200237393 1Attorney Docket No TGO-037WOmg / kg twice daily (BID). Vehicle was 5% DMA and 20% Captisol. Data are presented as mean + SEM.

[0033] FIG. 6A is a graph depicting the n vitro combination benefit of abemaciclib (Compound 2) and Compound 1 in U87MG MTAP-deleted glioblastoma (GBM) cell line as measured through cell titer Gio assay. FIG. 6B is a graph depicting results from colony formation assay to test combination of Compound 1 (TNG456 in FIG. 6B) and abemaciclib in U87MG MTAP-deleted GBM cell line. * refers to p-value = 0.0454 for abemaciclib vs TNG456 + abemaciclib and p-value = 0.0176 for TNG456 vs TNG456 + abemaciclib. FIG.6C is a Kaplan meier survival curve in an MTAP-null GBM (U87MG) orthotopic CDX model treated with 30 mg / kg BID Compound 1 (TNG456 in FIG. 6C) as a single agent or in combination with abemaciclib dosed at 50 mg / kg QD. * refers to p-value <0.05 and ** refers to p-value <0.01 according to log-rank test (Mantel-Cox test).

[0034] FIGs. 7A-7D are graphs depicting body weight change (%) (FIGs. 7A and 7C) and antitumor activity (FIGs. 7B and 7D) of Compound 1 (TNG456 in FIGs. 7A-7D) in combination with abemaciclib in MTAP-null human glioblastoma xenograft model (AM38). Compound 1 was dosed at 45 mg / kg BID (FIGs. 7A-7B) or 90 mg / kg BID (FIGs. 7C-7D) and 50 mg / kg once daily (QD) and abemaciclib was dosed at 50 mg / kg QD (FIGs. 7A-7B) or 20 mg / kg QD (FIGs. 7C-7D). Vehicle for TNG456: 5% DMA+20% Captisol; vehicle for abemaciclib: 1% hydroxyethyl cellulose + 0.1% antifoam in 25 mM PB pH 2; vehicle for combination was 5% DN and 20% Captisol co-dosed with 1% hydroxy ethyl cellulose + 0 1% antifoam in 25 mM PB pH 2. Data are presented as mean + SEM.

[0035] FIGs. 8A-8B are graphs depicting body weight change (%) (FIG. 8A) and antitumor activity (FIG. 8B) of Compound 1 (TNG456 in FIGs. 8A-8B) in an ST458 MTAP-Null PDX Model. Compound I was dosed at 30 mg / kg, 60 mg / kg and 90 mg / kg BID Vehicle 5% DMA+20% Captisol. Data are presented as mean + SEM.

[0036] FIG. 9 is a schematic of phase 1 / 2 clinical study for testing Compound 1 (TNG456 in FIG. 9) as a single agent or m combination with abemaciclib.

[0037] FIG. 10 is a graph depicting Plasma and CSF concentrations of TNG456 following two oral administrations given 8 hours apart in male cynomolgus monkeys.DETAILED DESCRIPTION

[0038] The disclosure herein sets forth exemplary methods, parameters and the like. It should be recognized, however, that such description is not intended as a limitation on the 10IPTS- 200237393 1Attorney Docket No TGO-037WOscope of the present disclosure but is instead provided as a description of exemplary7embodiments.Definitions

[0039] As used in the present disclosure, the following words and phrases are generally intended to have the meanings as set forth below unless expressly indicated otherwise or the context in which they are used indicates otherwise.MTAP

[0040] ''MTAP” as used herein refers to methylthioadenosine phosphorylase, an enzyme in the methionine salvage pathway, also known as S-methyl-5'-thioadenosine phosphorylase; also known as BDMF; DMSFH: DMSMFH; LGMBF: MSAP: and c86fus. External IDs:0MIM: 156540 MGI: 1914152 HomoloGene: 1838 chEMBL: 4941 GeneCards: MTAP Gene; Entrez 4507; RefSeq (mRNA): NM_002451; location: Chr 9: 21.8-21.93 Mb. By “wild-type” MTAP is meant that encoded by NM_002451 or having the same amino acid sequence (NP_002442). (Schmid et al. Oncogene 2000. 19. pp 5747-54).

[0041] As used herein, the term ‘" MTAP-deficienf ’,‘’MTAP-deficiency”,“MTAP-null” and the like refer to cells (including, but not limited to, cancer cells, cell lines, tissues, tissue types, tumors, etc.) that have a significant reduction in post- translational modification, production, expression, level, stability and / or activity of MTAP relative to that in a control, e.g. reference or normal or non-cancerous cells. The reduction can be at least about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80% or about 90%. In some embodiments, the reduction is at least about 20%, In some embodiments, the reduction is at least about 50%. Ihe terms “MTAP-deficient and / or MTA accumulating”. “MTAP-deficient and / or MTA-accumulatmg”, MTAP deficient and / or MTA upregulated” and the like, regarding a cell or cells, etc, indicate that the cell or cells, etc., either are deficient in MTAP and / or overproduce or accumulate MTA. MTAP-deficient cells include those wherein the MTAP gene has been mutated, deleted, or transcriptionally silenced. As a non-limiting example. MTAP-deficient cells can have a homozygous deletion. MTAP knockdown is not lethal. In some embodiments, the MTAP-deficient cells are also cyclin-dependent kinase inhibitor 2A (CDKN2A)-deficient. The MTAP deficiency can be detected using any reagent or technique known in the art, for example: immunohistochemistry utilizing an antibody to MTAP. and / or genomic sequencing, and / or nucleic acid hybridization and / or amplification 11IPTS- 200237393 1Attorney Docket No TGO-037WOutilizing at least one probe or primer comprising a sequence of at least 12 contiguous nucleotides (nt) of the sequence of MTAP, wherein the primer is no longer than about 30 nt.

[0042] An “MT AP-deficiency -related'’ or “MTAP-deficiency” or “MTAP deficient” disease (for example, a proliferating disease, e.g., a cancer) or a disease (for example, a proliferating disease, e.g, a cancer)“associated with MTAP deficiency” or a disease (for example, a proliferating disease, e.g., a cancer) “characterized by MTAP deficiency” and the like refer to an ailment (for example, a proliferating disease, e.g., a cancer) wherein a significant number of cells are MTAP-deficient. For example, m a MTAP-deficiency-related disease, one or more disease cells can have a significantly reduced post-translational modification, production, expression, level, stability and / or activity of MTAP. Examples of MTAP-deficiency-related diseases include, but are not limited to, cancers, including but not limited to: glioma (e.g., glioblastoma), cancers with high risk of brain metastasis, malignant peripheral nerve sheath tumors (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 or lung adenocarcinoma), astrocytoma, undifferentiated pleiomorphic sarcoma, diffuse large B-cell lymphoma (DI.. BCL), leukemia, head and neck cancer, stomach adenocarcinoma, myxofibrosarcoma, cholangiocarcinoma, cancer of the brain, stomach cancer, kidney cancer, breast cancer, endometrium cancer, urinary’ tract cancer, liver cancer, soft tissue cancer, pleura cancer, large intestine cancer or sarcoma. In some embodiments, the MTAP-deficiency-related diseases include glioma (e.g.. glioblastoma) non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma.

[0043] In a patient afflicted with a MTAP-deficiency-related disease, it is possible that some disease cells e.g., cancer cells) can be MTAP-deficient while others are not. Similarly, some disease cells may be MTA-accumulating while others are not. Thus, the present disclosure encompasses methods of treatment involving diseases of these tissues, or any other tissues, wherein the proliferation of MTAP-deficient and / or MTA-accumulating cells can be inhibited by administration of a PR. MT5 inhibitor. Some cancer cells which are MTAP-deficient are also deficient in CDKN2A; the post-translational modification, production, expression, level, stability’ anchor activity of the CDKN2A gene or its product are decreased in these cells. The genes for MTAP and CDKN2A are in close proximity on chromosome 9p21; MTAP is located approximately 100 kb telomeric to CDKN2A. Many cancer cell types 12IPTS / 200237393 1Attorney Docket No TGO-037WOharbor CDKN2A / MTAP loss (loss of both genes). Thus, in some embodiments, aMTAP-defi cient cell is also deficient in CDKN2A.MT A and MT A accumulation

[0044] By “MTA” is meant the PRMT5 inhibitor also known as methyl-thioadenosine, S-methyl-5’-thioadenosine, [5'deoxy-5'-(meihylthio)-fl-D-ribofuranosyl] adenine, 5'-methyl- thioadenosme, 5'-deoxy, 5'-methyl thioadenosine. and the like. MTA selectively inhibits PRMT5 methyltransferase activity. MTA is the sole known catabolic substrate for MTAP. The terms ‘ MTA accumulating”, ‘’MTA overproducing”, “MTA upregulated” and the like refer to cells (including, but not limited to, cancer cells, cell lines, tissues, tissue types, tumors, etc.) that have a significantly increased production, level and / or stability of MTA. MTA-accumulating cells include those wherein the cells comprise at least about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or greater than 100%, higher production, level and / or stability of MIA than that in normal or non-cancerous cells. In some embodiments, MTA-accumulating cells include those wherein the cells comprise at least 20% higher production, level and / or stability of MTA than that in normal or non-cancerous cells. In some embodiments. MTA-accumulating cells include those wherein the cells comprise at least 50% higher production, level and / or stability of MTA than that in normal or non-cancerous cells. Determination of MTA accumulation in test samples (e.g., cells such as cancer cells being tested for MTA accumulation) and reference samples, and other cells, tissues, samples, etc., can be performed using any method known in the art. Such methods for detecting MTA include, as a non-limiting example, liquid chromatography -electrospray ionization-tandem mass spectrometry7(LC-ESI-MS / MS), as described in Stevens et al. J. Chromatogr. A. 2010, 1217. pp 3282-3288; and Kirovski et al. Am. J. Pathol. 201 1, 178, pp 1145-1152; and references cited therein. Loss of MTAP is associated with accumulation of MTA (Williams-Ashman et al. Biochem. Pharm. 1982, 31, pp 277-288; and Limm et al. Eur. J Cancer. 2013, 49, Issue 6.

[0045] An “MTA-accumulation-related”, “MTA-accumulation”, “MTA-accumulating”, “MTA overproducing”, “MTA upregulated” disease (for example, a proliferating disease, e.g, a cancer) or a disease (for example, a proliferating disease, e.g, a cancer) “associated with MTA accumulation” or a disease (for example, a proliferating disease, e.g, a cancer) “characterized by MTA accumulation” and the like refer to an ailment (for example, a proliferating disease, e.g., a cancer) wlierein a significant number of cells are MTA13IPTS / 200237393 1Attorney Docket No TGO-037WOaccumulating Examples of MTA-accumulating diseases include, but are not limited to, cancers, including but not limited to: glioma (e.g., glioblastoma), cancers with high risk of brain metastasis, malignant peripheral nerve sheath tumors (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 or lung adenocarcinoma), astrocytoma, undifferentiated pleiomorphic sarcoma, diffuse large B-cell lymphoma (DLBCL), leukemia, head and neck cancer, stomach adenocarcinoma, myxofibrosarcoma, cholangiocarcmoma, cancer of the brain, stomach cancer, kidney cancer, breast cancer, endometrium cancer, urinary tract cancer, liver cancer, soft tissue cancer, pleura cancer, large intestine cancer or sarcoma. In some embodiments, the MTA-accumulating disease is glioma (e.g., glioblastoma) non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma.

[0046] In a patient afflicted with a MTAP-deficiency-related disease, it is possible that some disease cells (e.g., cancer cells) can be MTAP-deficient while others are not.In a patient having or having been diagnosed with an MTA-accumulating disease, some cells may be MTA-accumulating while others are not.

[0047] An increase in therapeutic window' between normal cells and MTAP-deleted / MTA accumulating cells could be achieved by using an inhibitor that binds PRMT5 uncoinpetitively with MTA. As used herein, “uncompetitive binding” and “uncompetitive inhibition” and “cooperative binding” and “cooperative inhibition” (e.g., MTA-uncompetitive binding, MTA-uncompetitive inhibition. MTA-cooperative binding, MTA-cooperative inhibition) refers to binding of an inhibitor to a protein (e.g, PRMT5) that is increased in the presence of a co-factor (e.g., MTA) over the binding of the same inhibitor in the absence of the co-factor. The PRMT5 inhibitors known in the art are generally either SAM (S-adenosylmethionine) uncompetitive or SAM competitive. As the concentration of SAM in wild-type and MTAP-null cells is similar, these inhibitors are expected to bind with similar potency to both cell types. By contrast, an MTA-cooperative (and either SAM competitive or showing enhanced cooperativity with MTA relative to SAM) inhibitor 'ould bind with apparent greater potency in the presence of high concentrations of MTA and w ould therefore result in preferential inhibition of PRMT5 m MTA-accumulating cells relative to normal cells.14IPTS / 200237393 1Attorney Docket No TGO-037WO

[0048] As described further herein, a cancer cell, a cancer type, or a subject with cancer, is “PRMT5 inhibitor sensitive,” sensitive to treatment with PRMT5 inhibitors,” sensitive to PRMT5 therapeutic inhibition,” or described in similar terms if it is amenable to treatment with a PRMT5 inhibitor, eg, due to its MTAP deficiency and / or MT A accumulation character.PRMT5

[0049] ‘“PRMT5” as used herein is the gene or protein Protein Arginine Methyltransferase 5, also known as HRMT1L5; IBP72: JBP1; SKB1; or SKBlIIs External IDs: OMIM:604045, MGI: 1351645, HomoloGene: 4454, ChEMBL: 1795116, GeneCards: PRMT5 Gene; EC number 2.1.1.125. Ensembl ENSG00000100462; UniProt 014744; Entrez Gene ID: 10419; RefSeq (mRNA): NM__001039619. The mouse homolog is NM__013768.Methyltransferases such as PRMT5 catalyze the transfer of one to three methyl groups from the co-factor S-adenosylmethionine (also known as SAM or AdoMet) to lysine or arginine residues of histone proteins. Argmine methylation is carried out by 9 different protein arginine methyltransferases (PRMT) in humans. Three types of methylarginine species exist: (I) Monomethylarginine (MMA); (2) Asymmetric dimethyl arginine (ADMA), which is produced by Type 1 methyl transferases (PRMTl, PRMT2, PRMT3, CARMI, PRMT6 and PRMT8); and (3) Symmetrical dimethylarginine (SDMA), which is produced by Type II methyl transferases (PRMT5 and PRMT7). PRMTl and PRMT5 are the major asymmetric and symmetric argmine methyltransferases, respectively, PRMT5 promotes symmetric dimethylation on histones at H3R8 and H4R3 (H4R3me2). Symmetric methylation of H4R3 is associated with transcriptional repression and can act as a binding site for DNMT3A. Loss of PRMT5 results in reduced DNMT3A binding and gene activation. Tumor suppressor gene ST7 and chemokines RANTES, IP 10, CXCL11 are targeted and silenced by PRMT5. WO 2011 / 079236.

[0050] Additional substrates include E2F1, p53, EGFR and CRAF. PRMT5 is part of a multi-protein complex comprising the co-regulatory factor WDR77 (also known as MEP50, a CDK4 substrate) during Gl / S transition. Phosphorylation increases PRMT5 / WDR77 activity. WDR77 is the non-catalytic component of the complex and mediates interactions with binding partners and substrates. PRMT5 can also interact with pICIn or RioKl adaptor proteins in a mutually exclusive fashion to modulate complex composition and substrate specificity.15IPTS / 200237393 1Attorney Docket No TGO-037WO

[0051] PRMT5 has either a positive or negative effect on its substrates by arginine methylation when interacting with a number of complexes and is involved in a variety of cellular processes, including RNA processing, signal transduction, transcriptional regulation, and germ cell development. PRMT5 is a major pro-survival factor regulating elF4E expression and p53 translation. PRMT5 triggers p53-dependent apoptosis and sensitized various cancer cells to Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) without affecting TRAIL resistance in non-transformed cells.

[0052] The term PRMT5 inhibitor" refers to any compound capable of inhibiting the production, level, activity, expression or presence of PRMT5. These include, as non-limiting examples, any compound inhibiting the transcription of the gene, the maturation of RNA, the translation of mRNA, the posttranslational modification of the protein, the enzymatic activity of the protein, the interaction of same with a substrate, etc. The term also refers to any agent that inhibits the cellular function of the PRMT5 protein, either by ATP-competitive inhibition of the active site, allosteric modulation of the protein structure, disruption of protein-protein interactions, or by inhibiting the transcription, translation, post-translational modification, or stability of PRMT5 protein.

[0053] In some embodiments, a PRMT5 inhibitor competes with another compound, protein or other molecule which interacts with PRMT5 and is necessary for PRMT5 function. As a non-limiting example, a PRMT5 inhibitor can compete with the co-factor S-adenosylmetliionine (also known as SAM or AdoMet).In some embodiments, the PRMT5 inhibitor is uncompetitive with MTA. In some embodiments, the PRMT5 inhibitor is uncompetitive with MTA and competitive with SAM, In some embodiments, the PRMT5 inhibitor is uncompetitive with MTA and uncompetitive with SAM but binds with a higher degree of potency for tine MTA complex relative to the S M complex.CDK4 / 6

[0054] CDK4’’ as used herein is the gene or protein cyclin-dependent kinase 4, also known as PSK-J3, ceil division protein kinase 4, and CMM3, External IDs: HGNC: 1773, NCBI Gene: 1019. Ensembl: ENSG00000135446, OMIM’®: 123829. UniProtKB / Swiss-Prot:Pl 1802, and RefSeq (mRNA): NM__000075.4. The mouse homolog is NM_009870.4 (isoform 1) and NM_001355005,1 (isoform 2). The protein encoded by the CDK4 gene is a16IPTS / 200237393 1Attorney Docket No TGO-037WOmember of the Ser / Thr protein kinase family, which forms a complex with cyclin D to regulate cell cycle Gl / S phase transition.

[0055] “CDK6” as used herein is the gene or protein cyclin-dependent kinase 6, also known as PLSTIRE. cell division protein kinase 6, Serine / Threonine-Protein Kinase PLSTIRE, CDKN6. External IDs: HGNC: 1777, NCB1 Gene: 1021, Ensembl: ENSG00000105810, OMIM*: 603368, UniProtKB / Swiss-Prot: Q00534, and RefSeq (mRNA): NM_001259.8 (transcript variant 1) and NM_001145306.2 (transcript variant 2), The mouse homolog is NM 009873.3. The protein encoded by the CDK6 gene is a member of the Ser / Thr protein kmase family, which forms a complex with cyclin D to regulate progression through the G1 phase of the cell cycle and the Gl / S phase transition.

[0056] The term “CDK4 / 6 inhibitor” refers to any compound capable of inhibiting the production, level, activity', expression or presence of CDK4 and / or CDK6. These include, as non-limiting examples, any compound inhibiting the transcription of the gene, the maturation of RNA, the translation of mRNA, the posttranslational modification of the protein, the enzy matic activity of the protein, the interaction of same with a substrate, etc. The term also refers to any agent that inhibits the cellular function of tire CDK4 and'or CDK6 protein, either by ATP-competitive inhibition of the active site, allosteric modulation of the protein structure, disruption of protein-protein interactions, or by inhibiting the transcription. translation, post-translational modification, or stability of the CDK4 and / or CDK6 protein. In some embodiments, the CDK4 / 6 inhibitor inhibits the catalytic activity7of CDK4 in complex with cyclin D.Other definitions

[0057] As used herein, the terms “a” and “an” mean “one or more” and include the plural unless the context is inappropriate.

[0058] As used herein, the term “salt” refers to any- and all salts and encompasses pharmaceutically acceptable salts.

[0059] The term “pharmaceutically7acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity7, irritation, allergic response and the like, and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al., describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the 17IPTS / 200237393 1Attorney Docket No TGO-037WOcompounds of this disclosure include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the ail such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate. digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemi sulfate. heptanoate, hexanoate, hydroiodide, 2-hydro.xy-ethanesulfonate, lactobionate, lactate, laurate, laury l sulfate, malate, maleate, malonate, metlianesulfonate. 2- naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonaie. undecanoate, valerate salts, and the like. Pharmaceutically acceptable salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(C i 4alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate.

[0060] A “subject’’ to which administration is contemplated includes, but is not limited to, humans (e.g. a male or female of any age group, e.g.. a pediatric subject (e.g., infant, child, adolescent) or adult subject (e.g., young adult, middle-aged adult or senior adult)) and / or a non-human animal, e.g., a mammal such as primates (e.g., cynomolgus monkeys, rhesus monkeys), cattle, pigs, horses, sheep, goats, rodents, cats, and / or dogs. In certain embodiments, the subject is a human. In certain embodiments, the subject is a non-human animal. The terms “human,” “patient,” and “subject” are used interchangeably herein. In some embodiments the subject is an adult human (e.g., > 18 years of age). In other embodiments the subject is a human child (e.g., < 18 years of age). In still other embodiments, the subject is a human female (adult or child). In yet other embodiments, the subject is a human male (adult or child).

[0061] Disease, disorder, and condition are used interchangeably herein.18IPTS / 200237393 1Attorney Docket No TGO-037WO

[0062] As used herein, and unless otherwise specified, the terms ‘’treat,” ‘’treating” and “treatment” contemplate an action that occurs while a subject is suffering from the specified disease, disorder or condition, which reduces the severity of the disease, disorder or condition, or retards or slows the progression of the disease, disorder or condition (“therapeutic treatment”), and also contemplates an action that occurs before a subject begins to suffer from the specified disease, disorder or condition (“prophyl ctic treatment”). In one embodiment, the compounds provided herein are contemplated to be used in methods of therapeutic treatment wherein the action occurs while a subject is suffering from the specified disease, disorder or condition and results in a reduction in the severity of the disease, disorder or condition, or retardation or slowing of the progression of the disease, disorder or condition. In an alternate embodiment, the compounds provided herein are contemplated to be used in methods of prophylactic treatment wherein the action occurs before a subject begins to suffer from the specified disease, disorder or condition and results in preventing a disease, disorder or condition, or one or more symptoms associated with the disease, disorder or condition, or preventing the recurrence of the disease, disorder or condition.

[0063] In general, the “effective amount” of a compound refers to an amount sufficient to elicit the desired biological response e.g., to treat a disease or disorder described herein. As will be appreciated by those of ordinary skill in this art, the effective amount of a compound of the disclosure may vary depending on such factors as the desired biological endpoint, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the age, health, and condition of the subject. An effective amount encompasses therapeutic and prophylactic treatment (z.e., encompasses a “therapeutically effective amount” and a “prophylactically effective amount”).

[0064] As used herein, and unless otherwise specified, a “therapeutically effective amount” of a compound is an amount sufficient to provide a therapeutic benefit in the therapeutic treatment of a disease, disorder or condition, or to delay or minimize one or more symptoms associated with the disease, disorder or condition. / X therapeutically effective amount of a compound means an amount of therapeutic agent, alone or in combination with other therapies, which provides a therapeutic benefit in the therapeutic treatment of the disease, disorder or condition. The term “therapeutically effective amount” can encompass an amount that improves overall therapy, reduces or avoids symptoms or causes of disease or condition, or enhances the therapeutic efficacy of another therapeutic agent. In some embodiments, the19IPTS / 200237393 1Attorney Docket No TGO-037WO"‘term "therapeutically effective amount” refers to a synergistically effective amount or synergistically therapeutic amount.

[0065] " Synergistic" means that the therapeutic effect of the PRMT5 inhibitor (e.g..Compound 1) when administered in combination as described herein with CDK4 / 6 inhibitor (e.g., Compound 2) is greater than the predicted additive therapeutic effects of the PRMT5 inhibitor and CDK4 / 6 inhibitor when administered alone at the same dose. The ‘‘term "synergistically therapeutic amount” or "synergistically effective amount” refers to a less than standard therapeutic amount of one or both drugs, meaning that the amount required for the desired effect is lower than when the drug is used alone. A synergistically therapeutic amount also includes cases where one drug is given at a standard therapeutic dose and another drag is administered in a less than standard therapeutic dose. For example, the PRMT5 inhibitor could be given in a therapeutic dose and the CDK4 / 6 inhibitor could be given in a standard or less than standard therapeutic dose to provide a synergistic result, or vice versa.

[0066] As used herein, and unless otherwise specified, a “prophylactically effective amount” of a compound is an amount sufficient to prevent a disease, disorder or condition, or one or more symptoms associated with the disease, disorder or condition, or prevent its recurrence. A prophylactically effective amount of a compound means an amount of a therapeutic agent, alone or in combination with other agents, which provides a prophylactic benefit in the prevention of the disease, disorder or condition. The term “prophylactically effective amount” can encompass an amount that improves overall prophylaxis or enhances the prophylactic efficacy of another prophylactic agent.

[0067] Where the use of the term “about” is before a quantitative value, the present invention also includes the specific quantitative value itself, unless specifically stated otherwise. As used herein, the term “about” refers to a ± 10% variation from the nominal value unless otherwise indicated or inferred.Compounds of the Present DisclosureCompound 1[00681 Provided herein are methods of treatment using a compound having the structural formula:20IPTS / 200237393 1Attorney Docket No TGO-037WOF (Compound 1); wherein the compound can be administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0069] Compound 1 is also known as TNG456 or the chemical name of (R)-N1-(4-amino- lH-pyrazolo[4,3-c]pyridin-7-yl)-N2-methyl-N2-(l-(4-(trifluoromethyl)phenyl)ethyl)oxal amide.

[0070] Compound 1 is designed to kill MTAP-deficient (e.g., MTAP-deleted) cancer cells while sparing normal cells. Compound 1 functions as a potent and selective PRMT5 inhibitor with MT A -cooperative binding, exhibiting a biochemical Ki,app of approximately 30 pM in the absence of MTA and approximately 2-2.5 pM in the presence of MTA. This MTA-cooperative mechanism underlies the selectivity of Compound 1 for MTAP-deleted tumors, which accumulate high levels of endogenous MTA.

[0071] In cellular assays. Compound 1 demonstrates a half-maximal inhibitory concentration (IC50) of approximately 2 nM in pharmacodynamic studies and a growth inhibition 50% (GI50) of approximately 20 nM in MT AP -null cancer cell lines. Compound 1 exhibits approximately 55-fold selectivity for MT AP -null versus MTAP wild-type cells across isogenic cell line pairs, demonstrating a favorable therapeutic window.[00721 Compound 1 is able to cross the blood- brain barrier (BBB) and penetrate the central nervous system (CNS). The brain penetrance of Compound 1 has been characterized by an unbound brain-to-plasma partition coefficient (Kp,uu) ranging from 0.45 to 1.1 as measured in non-human primates and dogs, indicating effective CN S distribution. This brain-penetrant property makes Compound 1 particularly suitable for treating CNS tumors, including glioblastoma, as well as advanced or metastatic solid tumors with CNS involvement or high risk of brain metastasis

[0073] The (R)-enantiomer configuration of Compound 1 demonstrates approximately 15-fold greater potency than the corresponding (S)-enantiomer.

[0074] Compound 1 exhibits minimal off-target activity across a panel of 40 methyltransferases at 10 pM concentration, with selective inhibition limited to the PRMT5 / MEP50 complex, demonstrating high target specificity.Attorney Docket No TGO-037WO

[0075] Throughout the embodiments and claims provided herein. Compound 1 and TNG456 are used interchangeably. Compound 1 can be prepared by the procedures described in PCT publication WO2023146987.2

[0076] Pro vided herein are methods of treatment using Compound 1 in combination with a second therapeutic agent, which therapeutic agent is a CDK4 / 6 inhibitor and having the structural formula:FCompound 2; wherein Compound 2 can be administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0077] Compound 2 is also known as abemaciclib. Abemaciclib is marketed, with Verzenio* as one of its tradenames. Abemaciclib is an inhibitor of cyclin-dependent kinases 4 and 6 (CDK4 and CDK6). The chemical name for abemaciclib is 2-Pyrimidinamine, N-[5-[(4-ethyl-1-piperazinyl)methyl]-2-pyridinyl]-5-fluoro-4-[4-fluoro-2-methyl-1-(1-methylethyl)-1H-benzimidazol-6-yl]-. Abemaciclib has the following structure:Abemaciclib (Compound 2)

[0078] Compound 2 can be prepared following the procedures outlined in U. S, Patent No, 7,855,211.

[0079] Abemaciclib can be formulated for oral administration. For example, abemaciclib can be provided as immediate-release oval white, beige, or yellow tablets. Inactive ingredients are as follows: Excipients — microcrystalline cellulose 102, microcrystalline cellulose 101, lactose monohydrate, croscarmellose sodium, sodium stearyl fumarate, silicon dioxide Color mixture ingredients — polyvinyl alcohol, titanium dioxide, polyethylene glycol, talc, iron oxide yellow', and iron oxide red.

[0080] Abemaciclib can be used as a monotherapy for the treatment of adult patients with hormone receptor (HR)-positive. human epidermal growth factor receptor 2 (HER2)-negative advanced or metastatic breast cancer with disease progression following endocrine therapy and prior chemotherapy in the metastatic setting. It can also be used in combination with an 22IPTS'200237393.1Attorney Docket No TGO-037WOendocrine therapy (e.g., fulvestrant, tamoxifen, or an aromatase inhibitor) to treat HRpositive, HER2 -negative breast cancer. For example, a combination therapy of abemaciclib and fulvestrant can be used for the treatment of HR-positive, HER2 -negative advanced or metastatic breast cancer with disease progression following endocrine therapy.

[0081] Recommended starting dose for abemaciclib as monotherapy is 200 mg twice daily. Recommended starting dose for abemaciclib in combination with other agents (e.g., fulvestrant, tamoxifen, or an aromatase inhibitor) is 150 mg twice daily.Pharmaceutical Compositions

[0082] The term '‘pharmaceutically acceptable carrier or adjuvant” refers to a carrier or adjuvant that may be administered to a patient, together with a compound provided herewith, and which does not destroy the pharmacological activity thereof and is nontoxic when administered in doses sufficient to deliver a therapeutic amount of the compound

[0083] Pharmaceutically acceptable carriers, adjuvants and vehicles that may be used in the pharmaceutical compositions provided herewith include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug deli very systems (SEDDS) such as d-a-tocopherol polyethyleneglycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tweens or other similar polymeric deliver}' matrices, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene polyoxypropylene block polymers, polyethylene glycol and wool fat. Cyclodextrins such as a-, [3-, and y-cyclodextrin, or chemically modified derivatives such as hydroxyalkylcyclodextrins, including 2 and 3 hydroxypropyl-p-cyclodextrins, or other solubilized derivatives may also be advantageously used to enhance delivery of compounds of the formulae described herein.

[0084] Generally, the compounds provided herein are administered in an effective amount (e.g., a therapeutically effective amount). The amount of the compound actually administered will typically be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound23IPTS / 200237393 1Attorney Docket No TGO-037WOadministered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like.

[0085] The pharmaceutical compositions provided herewith may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally. vaginally or via an implanted reservoir, preferably by oral administration or administration by injection. The pharmaceutical compositions provided herewith may contain any conventional nontoxic pharmaceutically acceptable carriers, adjuvants or vehicles. In some cases, the pH of the formulation may be adjusted with pharmaceutically acceptable acids, bases or buffers to enhance the stability of the formulated compound or its delivery form. The term parenteral as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrastemal, intrathecal, intralesional and intracranial injection or infusion techniques.

[0086] The compositions for oral administration can take the form of bulk liquid solutions or suspensions, or bulk powders. More commonly, however, the compositions are presented in unit dosage forms to facilitate accurate dosing. The term ’‘unit dosage forms” refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient. Typical unit dosage forms include prefilled, premeasured ampules or syringes of the liquid compositions or pills, tablets, capsules or the like in the case of solid compositions, in such compositions, the compound is usually a minor component (from about 0.1 to about 50% by weight or preferably from about 1 to about 40% by weight) with the remainder being various vehicles or earners and processing aids helpful for forming the desired dosing form.

[0087] Liquid forms suitable for oral administration may include a suitable aqueous or nonaqueous vehicle with buffers, suspending and dispensing agents, colorants, flavors and the like. Solid forms may include, for example, any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or com starch; a lubricant such as magnesium stearate; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

[0088] Injectable compositions are typically based upon injectable sterile saline or phosphate-buffered saline or other injectable carriers known in the art. As before, the active 24IPTS / 200237393 1Attorney Docket No TGO-037WOcompound in such compositions is typically a minor component, often being from about 0.05 to 10% by weight with the remainder being the injectable carrier and the like. The pharmaceutical compositions may be in the form of a sterile injectable preparation, for example, as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to techniques known in the art using suitable dispersing or wetting agents (such as, for example, Tween 80) and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are mannitol, water, Ringer’s solution and isotonic sodium chloride solution. Tn addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono - or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically acceptable oils, such as olive oil or castor oil, especially in their polyoxy ethylated versions. These oil solutions or suspensions may also contain a long chain alcohol diluent or dispersant, or carboxymethyl cellulose or similar dispersing agents which are commonly used in the formulation of pharmaceutically acceptable dosage forms such as emulsions and or suspensions. Other commonly used surfactants such as Tweens or Spans and / or other similar emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.

[0089] Transdermal compositions are typically formulated as a topical ointment or cream containing the active ingredient(s). generally in an amount ranging from about 0.01 to about 20% by weight, preferably from about 0.1 to about 20% by weight, preferably from about 0.1 to about 10% by weight, and more preferably from about 0,5 to about 15% by' weight. When formulated as an ointment, the active ingredients will typically be combined with either a paraffinic or a water-miscible ointment base. Alternatively, the active ingredients may' be formulated in a cream with, for example an oil-in-water cream base. Such transdermal formulations are well-known in the art and generally include additional ingredients to enhance the dermal penetration of stability of the active ingredients or the formulation. All such known transdermal formulations and ingredients are included within the scope provided herein.25IPTS / 200237393 1Attorney Docket No TGO-037WO

[0090] The compounds provided herein can also be administered by a transdermal device. Accordingly, transdermal administration can be accomplished using a patch either of the reservoir or porous membrane type, or of a solid matrix variety.

[0091] The pharmaceutical compositions provided herewith may also be administered m the form of suppositories for rectal administration. These compositions can be prepared by mixing a compound provided herewith with a suitable non irritating excipient which is solid at room temperature but liquid at the rectal temperature and therefore will melt in the rectum to release the active components. Such materials include, but are not limited to. cocoa butter, beeswax and polyethylene glycols.

[0092] The pharmaceutical compositions provided herewith may be administered by nasal aerosol or inhalation Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavail ability, fluorocarbons, and / or other solubilizing or dispersing agents known in the art.

[0093] The above-described components for orally administrable, injectable or topically administrable, rectally administrable and nasally administrable compositions are merely representative. Other materials as well as processing techniques and the like are set forth in Part 8 of Remington 's Pharmaceutical Sciences. 17th edition. 1985, Mack Publishing Company, Easton, Pennsylvania, which is incorporated herein by reference.

[0094] The compounds described herein can also be administered in sustained release forms or from sustained release drug delivery systems, A description of representative sustained release materials can be found in Remingto ’s Pharmaceutical Sciences.

[0095] When the compositions provided herewith comprise a combination of Compound 1 and one or more additional therapeutic or prophylactic agents (c.g., Compound 2), both the compound and the additional agent should be present at dosage levels of between about 1 to 100%, and more preferably between about 5 to 95% of the dosage normally administered in a monotherapy regimen. The additional agents may be administered separately, as part of a multiple dose regimen, from the compounds provided herewith. Alternatively, those agents may be part of a single dosage form, mixed together with the compounds provided herewith m a single composition.

[0096] Also provided is the pharmaceutically acceptable acid addition salt or a composition of Compound 1.26IPTS / 200237393.1Attorney Docket No TGO-037WO

[0097] The acid which may be used to prepare the pharmaceutically acceptable salt is that which forms a non-toxic acid addition salt, i.e., a salt containing pharmacologically acceptable anions such as the hydrochloride, hydroiodide, hydrobromide, nitrate, sulfate, bisulfate, phosphate, acetate, lactate, citrate, tartrate, succinate, maleate, fumarate, benzoate, para-toluenesulfonate, and the like.

[0098] The compounds described herein can, for example, be administered by injection, intravenously, intraarterially, subdermally, intraperitoneally, intramuscularly, or subcutaneously; or orally, buccally, nasally, transmucosally, topically, in an ophthalmic preparation, or by inhalation, with a dosage ranging from about 0.5 to about 100 mg / kg of body weight, alternatively dosages between 1 mg and 1000 mg / dose, every 4 to 120 hours, or according to the requirements of the particular drug. The methods herein contemplate administration of an effective amount of compound or compound composition to achieve the desired or stated effect. Typically, the pharmaceutical compositions provided herewith will be administered from about 1 to about 6 times per day or alternatively, as a continuous infusion. Such administration can be used as a chronic or acute therapy. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. A typical preparation will contain from about 5% to about 95% active compound (w / w).Alternatively', such preparations contain from about 20% to about 80% active compound.

[0099] Lower or higher doses than those recited above may be required. Specific dosage and treatment regimens for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health status, sex, diet, time of administration, rate of excretion, drug combination, the severity and course of the disease, condition or symptoms, the patient’s disposition to the disease, condition or symptoms, and the judgment of the treating physician

[0100] Upon improvement of a patient's condition, a maintenance dose of a compound, composition or combination provided herewith may be administered, if necessary.Subsequently, the dosage or frequency of administration, or both, may be reduced, as a function of the symptoms, to a level at which the improved condition is retained when the symptoms have been alleviated to the desired level. Patients may. however, require intermittent treatment on a long term basis upon any recurrence of disease symptoms.Methods of Treatment and Use27IPTS / 200237393.1Attorney Docket No TGO-037WOTreatment of MTAP-deficient and / or MTA-accumulating proliferation disorders

[0101] 5-Methylthioadenosine phosphorylase (MTAP) catalyzes the reversible phosphorylation of S-methyl-5'-thioadenosine (MTA) to adenine and 5-methylthioribose-l-phosphate. MTAP-deletion is a common genetic event in human cancer. M I AP deletion frequency in a subset of human cancers is described in Cerami et al.. Cancer Discov.(2012);2(5):401-4; Gao et al., Sci. Signal. (2013);6(269):pll; and Lee et al., Nat. Gen. (2014) 46(11): 1227-32. For example, more than 50% of malignant peripheral nerve sheath tumor (MPNST) have deletions in MTAP (Lee et al., Nat. Gen. (2014)). Other cancers with high MTAP deletion frequencies are glioblastoma (GBM). mesothelioma, bladder cancer, pancreatic cancer, esophageal cancer, squamous lung cancer, melanoma, diffuse large B cell lymphoma (DLBCL), head and neck cancer, cholangiocarcinoma, lung adenoma, sarcoma, stomach cancer, glioma, adrenal carcinoma, thymoma, breast cancer, liver cancer, ovarian cancer, renal papillary cancer, uterine cancer, prostate cancer, and renal clear cell cancer MTAP deletion in cells is one of the mechanisms that leads to MTAP-deficiency, increased intracellular MTA accumulation, and confers enhanced dependency on the protein arginine methyltransferase 5 (PRMT5) in cancer cells. Other mechanisms leading to MTAP deficiency include, inter alia, MTAP translocations and MTAP epigenetic silencing which could also lead to MTAP -null and / or MTAP deficient tumors. PRMT5 mediates the formation of symmetric dimethylarginine (SDMA); thus, the PRMT5 activity can be assessed by measuring the SDMA levels using the antibody against an SDMA or SDMA modified polypeptide.

[0102] In one embodiment, provided are methods of treating human or animal subjects having or having been diagnosed with an MTAP-deficiency-related and / or MTA-accumulating proliferative disorder (e.g., cancer) comprising administering to the subject in need thereof a therapeutically effective amount of Compound 1 or a pharmaceutically acceptable salt or a composition thereof.[01031 In some embodiments, provided is Compound 1 or a pharmaceutically acceptable salt or a composition thereof for use in a method of treating human or animal subjects having or having been diagnosed with an MTAP-deficiency-related and / or MTA-accumulating proliferative disorder (e.g.. cancer). In some embodiments, Compound 1 is provided m a therapeutically effective amount.

[0104] In some embodiments, provided is Compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising Compound 1 or the28IPTS / 200237393 1Attorney Docket No TGO-037WOpharmaceutically acceptable salt, for use in the manufacturing of a medicament for treating human or animal subjects having or having been diagnosed with an MTAP-deficiency-related and / or MTA-accumulating proliferative disorder (e.g., cancer). In some embodiments.Compound 1 or the pharmaceutically acceptable salt or a composition thereof is provided in a therapeutically effective amount.

[0105] In some embodiments, provided is a use of Compound 1 or a pharmaceutically acceptable salt thereof or of a pharmaceutical composition comprising Compound 1 or the pharmaceutically acceptable salt, in a method of treating human or animal subjects having or having been diagnosed with an MTAP-deficiency-related and / or MTA-accumulating proliferative disorder (e.g., cancer). In some embodiments, the use is of a therapeutically effective amount of Compound 1 or the pharmaceutically acceptable salt thereof, or the composition comprising Compound 1 or the pharmaceutically acceptable salt.

[0106] In some embodiments, provided is use of Compound 1 or a pharmaceutically acceptable salt thereof, or of a pharmaceutical composition comprising Compound 1 or the pharmaceutically acceptable salt, in the manufacturing of a medicament for treating human or animal subjects having or having been diagnosed with an MTAP -deficiency-related and / or MTA-accumulating proliferative disorder (e.g., cancer). In some embodiments, the use is of a therapeutically effective amount of Compound 1 or the pharmaceutically acceptable salt, or a composition comprising Compound 1 or the pharmaceutically acceptable salt.

[0107] In one embodiment, provided are methods for treating an MTAP-deficiency-related and / or MTA-accumulating proliferative disorder (e.g., cancer) in a subject in need thereof comprising administering to the subject an effective amount (e.g., a therapeutically effective amount) of Compound 1 or a pharmaceutically acceptable salt thereof.

[0108] In one embodiment, provided are methods of treating human or animal subjects having or having been diagnosed with an MTAP-deficiency-related and / or MTA-accumulating proliferative disorder (e.g., cancer) comprising administering to the subject in need thereof a therapeutically effective amount of pharmaceutical composition of Compound 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutical composition is administered in combination with a second therapeutic agent.

[0109] In one embodiment, provided are methods of treating an MTAP-deficiency-related and / or MTA-accumulating proliferative disorder (e.g., cancer) in a subject in need thereof comprising administering to the subject a therapeutically effective amount of pharmaceutical 29IPTS / 200237393 1Attorney Docket No TGO-037WOcomposition comprising Compound 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutical composition is administered in combination with a second therapeutic agent.

[0110] In some embodiments, the subject is human.

[0111] In certain embodiments, the disease is an MTAP-deficient and / or MTA-accumulating cancer.

[0112] In one embodiment, the cancer is glioma (e.g., glioblastoma), cancers with high risk of brain metastasis, malignant peripheral nerve sheath tumors (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; eg., lung squamous or lung adenocarcinoma), astrocytoma, undifferentiated pleiomorphic sarcoma, diffuse large B-cell lymphoma (DLBCL), leukemia, head and neck cancer, stomach adenocarcinoma, myxofibrosarcoma, cholangiocarcinoma, cancer of the brain, stomach cancer, kidney cancer, breast cancer, endometrium cancer, urinary tract cancer, liver cancer, soft tissue cancer, pleura cancer, large intestine cancer or sarcoma. In some embodiments, the cancer is glioma (e.g., glioblastoma) non-small cell lung cancer, pancreatic cancer, bladder cancer or chol angi ©carcinoma.

[0113] In one embodiment, the cancer is an MTAP-deficient and / or MTA-accumulating glioma (e.g., glioblastoma), cancers with high risk of brain metastasis, malignant peripheral nerve sheath tumors (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 or lung adenocarcinoma), astrocytoma, undifferentiated pleiomorphic sarcoma, diffuse large B-cell ly mphoma (DLBCL), leukemia, head and neck cancer, stomach adenocarcinoma, myxofibrosarcoma, cholangiocarcinoma, cancer of the brain, stomach cancer, kidney cancer, breast cancer, endometrium cancer, urinary tract cancer, liver cancer, soft tissue cancer, pleura cancer, large intestine cancer or sarcoma. In some embodiments, the cancer is an MTAP-deficient and / or MTA-accumulating glioma (e.g., glioblastoma) non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma.

[0114] Provided herein is the use of a PRMT5 inhibitor having the structure of Compound 1 or a pharmaceutically acceptable salt thereof in a method of inhibiting proliferation of 30IPTS / 200237393.1Attorney Docket No TGO-037WOMTAP-deficient cells in a subject in need thereof, the method comprising the step of administering to the subject the PRMT5 inhibitor or a composition thereof in an amount that is effective to inhibit proliferation of the MTAP-deficient cells. In one embodiment, the subject in need thereof suffers from a cancer selected from the group consisting of glioma (e.g., glioblastoma), cancers with high risk of brain metastasis, malignant peripheral nerve sheath tumors (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 or lung adenocarcinoma), astrocytoma, undifferentiated pleiomorphic sarcoma, diffuse large B-cell lymphoma (DLBCL), leukemia, head and neck cancer, stomach adenocarcinoma, myxofibrosarcoma, cholangiocarcinoma, cancer of the brain, stomach cancer, kidney cancer, breast cancer, endometrium cancer, urinary tract cancer, liver cancer, soft tissue cancer, pleura cancer, large intestine cancer or sarcoma. In some embodiments, the subject in need thereof suffers from a cancer selected from the group consisting of glioma (e g., glioblastoma) non-small cell lung cancer, pancreatic cancer, bladder cancer and cholangiocarcinoma.

[0115] Provided herein is the use of a PRMT5 inhibitor having the structure of Compound I or a pharmaceutically acceptable salt thereof in a method of inhibiting proliferation of MTA-accumulating cells in a subject in need thereof, the method comprising the step of administering to the subject the PRMT5 inhibitor or a composition thereof in an amount that is effective to inhibit proliferation of the MTA-accumulating cells. In one embodiment, the subject in need thereof suffers from a cancer selected from the group consisting of glioma (e.g, glioblastoma), cancers with high risk of brain metastasis, malignant peripheral nerve sheath tumors (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 or lung adenocarcinoma), astrocytoma, undifferentiated pleiomorphic sarcoma, diffuse large B-cell lymphoma (DLBCL), leukemia, head and neck cancer, stomach adenocarcinoma, myxofibrosarcoma, cholangiocarcinoma, cancer of the brain, stomach cancer, kidney cancer, breast cancer, endometrium cancer, urinary tract cancer, liver cancer, soft tissue cancer, pleura cancer, large intestine cancer or sarcoma. In some embodiments, the subject in need thereof suffers from a cancer selected from glioma31IPTS / 200237393 1Attorney Docket No TGO-037WO(e.g., glioblastoma) non-small cell lung cancer, pancreatic cancer, bladder cancer and cholangiocarcinoma.

[0116] Provided herein is the use of a PRMT5 inhibitor having the structure of Compound 1 or a pharmaceutically acceptable salt thereof in a method of inhibiting proliferation of MTAP deficient and / or MTA-accumulating cells in a subject in need thereof, the method comprising the step of administering to the subject the PRMT5 inhibitor or a composition thereof in an amount that is effective to inhibit proliferation of the MTAP deficient and / or MTA-accumulating cells. In one embodiment, the subject in need thereof suffers from a cancer selected from the group consisting of glioma (e.g., glioblastoma), cancers with high risk of brain metastasis, malignant peripheral nerve sheath tumors (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 or lung adenocarcinoma), astrocytoma, undifferentiated pleiomorphic sarcoma, diffuse large B-cell lymphoma (DLBCL), leukemia, head and neck cancer, stomach adenocarcinoma, myxofibrosarcoma, cholangiocarcinoma, cancer of the brain, stomach cancer, kidney cancer, breast cancer, endometrium cancer, urinary tract cancer, liver cancer, soft tissue cancer, pleura cancer, large intestine cancer or sarcoma. In some embodiments, the subject in need thereof suffers from a cancer selected from the group consisting of glioma (e.g., glioblastoma) non-small cell lung cancer, pancreatic cancer, bladder cancer andcholangiocarcinoma,

[0117] In some embodiments, the present disclosure provides a method of treating a MTAP-deficiency-related or MTAP deficient disease using a compound having the structuralformula:F (Compound 1); wherein the compound is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0118] In one aspect, the present disclosure provides a method of treating a cancer in a subject in need thereof using a compound having the structural formula:32IPTS- 200237393 1Attorney Docket No TGO-037WOr---N.F (Compound 1); wherein the compound is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0119] In some embodiments, the cancer comprises a solid tumor with MTAP-deficiency or MTAP-deletion. In some embodiments, the cancer comprises glioma (e.g., glioblastoma) non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma In some embodiments, the cancer comprises non-small cell lung cancer. In some embodiments, the cancer comprises pancreatic cancer. In some embodiments, the cancer comprises bladder cancer. In some embodiments, the cancer comprises cholangiocarcinoma.

[0120] In another aspect, the present disclosure provides a method of treating a tumor in the central nervous system (CNS) in a subject in need thereof using a compound having theF— Fstructural formula:F (Compound 1); wherein the compound is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0121] In some embodiments, the tumor in the CNS is a glioma In some embodiments, the glioma is a glioblastoma. In some embodiments, the tumor in the CNS (eg., glioma or glioblastoma) is MTAP-deficient. In some embodiments, the tumor in the CNS (e.g., glioma or glioblastoma) is MTA-accumulating.

[0122] In some embodiments, Compound 1 is administered twice daily (BID).Combination therapies

[0123] In some embodiments, provided are methods of treatment of MTAP-deficient and / or MTA accumulating proliferative disorders (e.g., cancers) with a PRMT5 inhibitor having the structure of Compound 1 or a pharmaceutically acceptable salt thereof in combination with one or more therapeutic agents.IPTS / 200237393.1Attorney Docket No TGO-037WO

[0124] Compound 1 has been designed with low cytochrome P4503A4 (CYP3A4) time¬ dependent inhibition (TDI) risk, which is particularly advantageous for combination therapy applications. This favorable drug-drug interaction (DDI) profile enables safe co-administration with CYP3A4 substrate drugs, including Compound 2 (abemaciclib), which is a sensitive CYP3A4 substrate. The low CYP3A4 TDI risk of Compound 1 minimizes the potential for clinically significant pharmacokinetic interactions that could lead to elevated exposures and increased toxicity1of co-administered CYP3A4 substrates. This property1makes Compound 1 particularly suitable for combination regimens in treating MTAP-deficient cancers, including glioblastoma, where combination with CDK4 / 6 inhibitors such as abemaciclib provides synergistic therapeutic benefit

[0125] In some embodiments, provided are methods of treatment of MTAP-deficient and / or MT A accumulating proliferative disorders (e.g, cancers) with a PRMT5 inhibitor described herein (i.e.. Compound 1 or a pharmaceutically acceptable salt thereof) in combination with a second therapeutic agent. In specific embodiments, provided are methods of treatment of MTAP-deficient and / or MIA accumulating proliferative disorders (e.g., cancers) with the PRMT5 inhibitor in combination with a CDK4 / 6 inhibitor having the structure of Compound 2 or a pharmaceutically acceptable salt thereof. In further specific embodiments, provided are methods of treatment of MTAP-deficient and / or MTA accumulating proliferative disorders (e.g., cancers) with a composition (eg., pharmaceutical composition) comprising the PRMT5 inhibitor in combination with a composition (e.g., pharmaceutical composition) comprising the CDK4 / 6 inhibitor.

[0126] The term " Combination'’ refers to either a fixed combination in one dosage unit form, or a combined administration where a PRMT5 inhibitor descnbed herein (i.e, Compound 1 or a pharmaceutically acceptable salt thereof) and a combination partner (e.g., abemaciclib, also referred to as “Compound 2’’) may' be administered independently at the same time or separately within time intervals, especially where these time intervals allow that the combination partners show a cooperative, e.g, synergistic effect. The single components may be packaged in a kit or separately. The terms “co-administration” or “combined administration’’ or the like as utilized herein are meant to encompass administration of the selected combination partner to a single subject in need thereof (e.g, a patient), and are intended to include treatment regimens in which the agents are not necessarily administered by the same route of administration or at the same time. The term “pharmaceutical combination’’ as used herein means a product that results from the mixing or combining of 34IPTS / 200237393 1Attorney Docket No TGO-037WOmore than one therapeutic agent and includes both fixed and non -fixed combinations of the therapeutic agents. The term “fixed combination” means that the therapeutic agents, e.g., the PRMT5 inhibitor or a composition thereof and the CDK4 / 6 inhibitor or a composition thereof, are both administered to a patient simultaneously in the form of a single entity or dosage. The term “non-fixed combination” means that the therapeutic agents, e.g., the PRMT5 inhibitor or a composition thereof and the CDK4 / 6 inhibitor or a composition thereof, are both administered to a patient as separate entities either simultaneously, concurrently or sequentially with no specific time limits, wherein such administration provides therapeutically effective levels of the two compounds in the body of the patient. The latter also applies to cocktail therapy, e.g., the administration of three or more therapeutic agents.

[0127] The term "combination therapy" refers to the administration of two or more therapeutic agents to treat a therapeutic condition or disorder described in the present disclosure. Such administration encompasses co-administration of these therapeutic agents in a substantially simultaneous manner, such as in a single capsule having a fixed ratio of active ingredients. Alternatively, such administration encompasses co-administration in multiple, or in separate containers (e.g., tablets, capsules, powders, and liquids) for each active ingredient. Powders and / or liquids may be reconstituted or diluted to a desired dose prior to administration. In addition, such administration also encompasses use of each type of therapeutic agent in a sequential manner, either at approximately the same time or at different times,

[0128] In some embodiments, provided is a method of treating a disease or disorder (e.g.. cancer) comprising administering or coadministering, in any order, to a patient in need thereof aPRMT5 inhibitor described herein (z.<?., Compound 1 or a pharmaceutically acceptable salt thereof) and a CDK4 inhibitor, including, but not limited to, LEE011 or a CDK 4 / 6 inhibitor (e.g, palbociclib (Ibrance®). ribociclib (Kisqali®), and abemaciclib (Verzenio®). In some embodiments, provided is a method of treating a disease or disorder (e.g., cancer) comprising administering or coadministering, in any order, to a patient in need thereof a Compound 1 or a pharmaceutically acceptable salt thereof and abemaciclib (Verzenio®).

[0129] Some patients may experience nausea during and after administration of the PRMT5 inhibitors described herein and / or other anti-cancer agent(s); therefore, anti-emetics are used in preventing nausea (upper stomach) and vomiting In some embodiments, provided is a 35IPTS / 200237393 1Attorney Docket No TGO-037WOmethod of treating a disease or disorder (e.g, cancer) comprising administering or coadministering, in any order, to a patient in need thereof a PRMT5 inhibitor described herein (i.e.. Compound 1 or a pharmaceutically acceptable salt thereof) or a composition thereof and an anti-emetic (e.g, aprepitant (Emend®), ondansetron (Zofran®), granisetron HC1 (Kytril®), lorazepam (Ativan®), dexamethasone (Decadron®), prochlorperazine (Compazine®), casopitant (Rezonic® and Zunrisa®), and combinations thereof).

[0130] Medication to alleviate the pain experienced during the treatment period is often prescribed to make the patient more comfortable. In some embodiments, provided is a method of treating a disease or disorder (e.g, cancer) comprising administering or coadministering, in any order, to a patient in need thereof a PRMT5 inhibitor described herein (i.e., Compound 1 or a pharmaceutically acceptable salt thereof) or a composition thereof and an analgesic (e.g, an over-the-counter analgesic (e.g., Tylenol®), an opioid analgesic (e.g., hydrocodone / paracetamol or hydrocodone / acetaminophen (e.g., Vicodin®1), morphine (e.g, Astramorph® or Avinza®), oxycodone (e g, OxyContin® or Percocet®), oxymorphone hydrochloride (Opana®), fentanyl (e.g., Duragesic®))).

[0131] In an effort to protect normal cells from treatment toxicity and to limit organ toxi cities, cytoprotective agents (such as neuroprotectants, free-radical scavengers, cardioprotectors, anthracycline extravasation neutralizers, nutrients and the like) may be used as an adjunct therapy. In some embodiments, provided is a method of treating a disease or disorder (e.g., cancer) comprising administering or coadministering, in any order, to a patient in need thereof a PRMT5 inhibitor described herein (i.e., Compound 1 or a pharmaceutically acceptable salt thereof) or a composition thereof and a cytoprotective agent (e.g. Amifostme (Ethyol®), glutamine, dimesna (Tavocept®). mesna (Mesnex®), dexrazoxane (Zinecard® or Totect®). xaliproden (Xaprila®), and leucovorin (also known as calcium leucovorin, citrovorum factor and folinic acid)),

[0132] In some embodiments, the present disclosure provides a method of treating a cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a protein arginine methyltransferase 5 (PRMT5) inhibitor and a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor, wherein the PRMT5 inhibitor is a compound having the structural formula:Attorney Docket No TGO-037WOF Compound 1: wherein the CDK4 / 6 inhibitor is a compound having the structural formula:Compound 2; wherein Compound 1 and Compound 2 are each administered as a free base or a pharmaceutically acceptable salt or a composition thereof. The selection of free base or a pharamaceutically acceptable salt can be made independently for Compound 1 and Compound 2.

[0133] In some embodiments, the present disclosure provides a method of treating a tumor in the CNS in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a protein arginine methyltransferase 5 (PRMT5) inhibitor and a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor. wherein the PRMT5 inhibitor is a compound having the structural formula:F Compound 1: wherein the CDK4 / 6 inhibitor is a compound having the structural formula:FCompound 2; wherein Compound 1 and Compound 2 are administered as a free base or a pharmaceutically acceptable salt or aIPTS'200237393.1Attorney Docket No TGO-037WOcomposition thereof. The selection of free base or a pharamaceutically acceptable salt can be made independently for Compound 1 and Compound 2.

[0134] The above-mentioned compounds, which can be used in combination with a PRMT5 inhibitor as described herein, can be prepared and administered as described in the art, including, but not limited to, in the documents cited above.

[0135] In particular, compositions will either be formulated together as a combination therapeutic or administered separately.

[0136] In combination therapy, a PRMT5 inhibitor as described herein and other anti-cancer agent(s) may be administered either simultaneously, concurrently or sequentially with no specific time limits, wherein such administration provides therapeutically effective levels of the two compounds in the body of the patient.

[0137] In a preferred embodiment, the PRMT5 inhibitor (i.e., Compound 1 or a pharmaceutically acceptable salt thereof) or a composition thereof and the CDK4 / 6 inhibitor (e.g., Compound 2 or a pharmaceutically acceptable salt thereof) or a composition thereof are administered sequentially in any order by infusion or orally. The dosing regimen may vary7depending upon the stage of the disease, phy sical fitness of the patient, safety- profiles of the individual drugs, and tolerance of the individual drugs, as well as other criteria well-known to the attending physician and medical practitioner(s) administering the combination. In some embodiments, the PRMT5 inhibitor or the composition thereof may’ be administered within minutes, hours, days, or even weeks apart from the CDK4 / 6 inhibitor or the composition thereof depending upon the particular cycle being used for treatment. In addition, the cycle could include administration of one drug more often than the other during the treatment cycle and at different doses per administration of the drug.

[0138] In another embodiment, provided are kits that include a PRMT5 inhibitor as disclosed herein (i.e., Compound 1 or a pharmaceutically acceptable salt thereof) and a CDK4 / 6 inhibitor as disclosed herein (e.g., Compound 2 or a pharmaceutically acceptable salt thereof) are provided. Representative kits include (a) the PRMT5 inhibitor ) and (b) the CDK4 / 6 inhibitor, whereby such kit may comprise a package insert or other labeling including directions for administration,

[0139] In some embodiments of the methods of treatment descnbed herein the combination of the PRMT5 inhibitor (e.g. Compound 1) and CDK4 / 6 inhibitor (e.g.. Compound 2) is synergistic.38IPTS / 200237393 1Attorney Docket No TGO-037WOSelected embodiments

[0140] Embodiment 1. A protein arginine methyltransferase 5 (PRMT5) inhibitor for use m treating a tumor in the central nervous system (CNS) in a subject in need thereof, wherein the PRMT5 inhibitor is a compound having the structural formula:hk A / NHF Compound 1;wherein the compound is administered to the subj ect as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0141] Embodiment 2. The compound for use of embodiment 1, wherein the tumor in the CNS is a glioma.

[0142] Embodiment 3. The compound for use of embodiment 2, wherein the glioma is a glioblastoma.

[0143] Embodiment 4. The compound for use of any one of embodiments 1 to 3, wherein the tumor in the CNS is MTAP-deficient.

[0144] Embodiment 5. The compound for use of any one of embodiments 1 to 4, wherein the tumor in the CNS is MTA-accumulating.

[0145] Embodiment 6. A protein arginine methyltransferase 5 (PRMT5) inhibitor for use in treating a cancer in a subject m need thereof, wherein the PRMT5 inhibitor is a compound having the structural formula:H2NN HF~4~FF Compound 1;wherein the compound is administered to the subject as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0146] Embodiment 7. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of embodiment 6, wherein the cancer comprises a solid tumor with MTAP-deficiency.39IPTS / 200237393.1Attorney Docket No TGO-037WO

[0147] Embodiment 8. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of embodiment 6 or 7, wherein the cancer comprises glioma (e.g., glioblastoma), non¬ small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma.

[0148] Embodiment 9. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of embodiments 1 to 8, wherein the PRMT5 inhibitor is administered systemically.

[0149] Embodiment 10. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of embodiments 1 to 9, wherein the PRMT5 inhibitor is administered orally.

[0150] Embodiment 11. The protein arginine methyl transferase 5 (PRMT5) inhibitor for use of any one of embodiments 1 to 10, wherein the PRMT5 inhibitor is administered twice daily (BID).

[0151] Embodiment 12. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of embodiments 1 to 11, wherein the subject also receives a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor having the structural formula:F HN; '. •NN.' Compound 2;wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0152] Embodiment 13. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of embodiment 12, wherein the CDK 4 / 6 inhibitor is administered systemically.

[0153] Embodiment 14. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of embodiment 12 or 13. wherein the CDK 4 / 6 inhibitor is administered orally.

[0154] Embodiment 15. A protein arginine methyltransferase 5 (PRMT5) inhibitor for use m combination with a cyclin-dependent kinase (CDK) 4 / 6 inhibitor in treating a cancer in a subject in need thereof, wherein:the PRMT5 inhibitor is a compound having the structural formula:IPTS'200237393.1Attorney Docket No TGO-037WOF Compound 1:wherein Compound 1 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof; andthe CDK4 / 6 inhibitor is a compound having the structural formula:Compound 2;wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0155] Embodiment 16. The protein argmine metliyltransferase 5 (PRMT5) inhibitor for use of embodiment 15, wherein the cancer comprises a solid tumor with MTAP-deficiency.

[0156] Embodiment 17. The protein arginine meth ltransferase 5 (PRMT5) inhibitor of embodiment 15 or 16, wherein the cancer comprises glioma (e.g, glioblastoma), non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma.

[0157] Embodiment 18. A protein arginine methyltransferase 5 (PRMT5) inhibitor for use m combination with a cyclin-dependent kinase (CDK) 4 / 6 inhibitor in treating a tumor in the central nervous system (CNS) in a subject in need thereof, wherein:the PRMT5 inhibitor is a compound having the structural formula:F Compound 1;wherein Compound 1 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof; andthe CDK4 / 6 inhibitor is a compound having the structural formula:IPTS'200237393.1Attorney Docket No TGO-037WOFCompound 2;wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0158] Embodiment 19. The protein arginine methy ltransferase 5 (PRMT5) inhibitor for use of embodiment 18. wherein the tumor in the CNS is a glioma.

[0159] Embodiment 20. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of embodiment 19, wherein the glioma is a glioblastoma.

[0160] Embodiment 21. The protein arginine methyl transferase 5 (PRMT5) inhibitor for use of any one of embodi ments 18 to 20, wherein the tumor in the CNS is MTAP-deficient.

[0161] Embodiment 22. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of anv one of embodiments 18 to 21, wherein the tumor in the CNS is MTA-accumulating.

[0162] Embodiment 23. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of embodiments 15 to 22, wherein the PRMT5 inhibitor is administered systemically.

[0163] Embodiment 24. The protein arginine methyl transferase 5 (PRMT5) inhibitor for use of any one of embodiments 15 to 23, wherein the PRMT5 inhibitor is administered orally.

[0164] Embodiment 25. The protein argmine methyltransferase 5 (PRMT5) inhibitor for use of any one of embodiments 15 to 24, wherein the PRMT5 inhibitor is administered twice daily.

[0165] Embodiment 26. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of embodiments 15 to 25, wherein the CDK4 / 6 inhibitor is administered systemically.

[0166] Embodiment 27. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of embodiments 15 to 26, wherein the CDK 4 / 6 inhibitor is administered orally.

[0167] Embodiment 28. A method of treating a tumor in the central nervous system (CNS) m a subject in need thereof, the method compnsing administering to the subject a therapeutically effective amount of a protein arginine methyltransferase 5 (PRMT5) inhibitor, wherein the PRMT5 inhibitor is a compound having the structural formula:IPTS'200237393.1Attorney Docket No TGO-037WOF Compound 1:wherein the compound is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0168] Embodiment 29. The method of embodiment 28, wherein the tumor in the CNS is a glioma.

[0169] Embodiment 30. The method of embodiment 29, wherein the glioma is a glioblastoma.

[0170] Embodiment 31. The method of any one of embodiments 28 to 30, wherein the tumor in the CNS is MTAP-deficient.

[0171] Embodiment 32. The method of any one of embodiments 28 to 31, wherein the tumor in the CNS is MTA-accumulating.

[0172] Embodiment 33. A method of treating a cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a protein arginine methyltransferase 5 (PRMT5) inhibitor, wherein the PRMT5 inhibitor is a compound having the structural formula:F Compound 1;wherein the compound is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0173] Embodiment 34. The method of embodiment 33, wherein the cancer comprises a solid tumor with MTAP-deficiency.

[0174] Embodiment 35. The method of embodiment 33 or 34, wherein die cancer comprises glioma (e.g, glioblastoma), non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma.IPTS / 200237393.1Attorney Docket No TGO-037WO

[0175] Embodiment 36. The method of any one of embodiments 28 to 35, wherein the PRMT5 inhibitor is administered systemically.

[0176] Embodiment 37. The method of any one of embodiments 28 to 36, wherein the PRMT5 inhibitor is administered orally.

[0177] Embodiment 38. The method of any one of embodiments 28-37, wherein the PRMT5 inhibitor is administered twee daily (BID).

[0178] Embodiment 39. The method of any one of embodiments 28 to 38, wherein the method further comprises administering to the subject a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor having the structural formula:NCompound 2;wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0179] Embodiment 40. The method of any one of embodiments 28 to 38, w herein the subject also receives a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor having the structural formula:FH. K. N.. >1.F Compound 2;wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof

[0180] Embodiment 41. The method of embodiment 39 or 40, wherein the CDK4 / 6 inhibitor is administered systemically.

[0181] Embodiment 42. The method of any one of embodiments 39 to 41, wherein the CDK 4 / 6 inhibitor is administered orally.

[0182] Embodiment 43. A method of treating a cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a protein arginine methyltransferase 5 (PRMT5) inhibitor and a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor,wherein the PRMT5 inhibitor is a compound having the structural formula:IPTS'200237393.1Attorney Docket No TGO-037WOF Compound 1:wherein the CDK4 / 6 inhibitor is a compound having the structural formula:wherein Compound 1 and Compound 2 are each administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0183] Embodiment 44. The method of embodiment 43, wherein the cancer comprises a solid tumor with MTAP-deficiency.

[0184] Embodiment 45. The method of embodiment 43 or 44, wherein the cancer comprises glioma (e.g., glioblastoma), non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcmoma.

[0185] Embodiment 46. A method of treating a tumor m the CNS in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a protein arginine methyl transferase 5 (PRMT5) inhibitor and a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor,wherein the PRMT5 inhibitor is a compound having the structural formula:F Compound 1:wherein the CDK4 / 6 inhibitor is a compound having the structural formula:45IPTS'200237393.1Attorney Docket No TGO-037WOwherein Compound 1 and Compound 2 are each administered as a free base or a pharmaceutically acceptable salt or a composition thereof

[0186] Embodiment 47. The method of embodiment 46, wherein the tumor in the CNS is a glioma.

[0187] Embodiment 48. The method of embodiment 47, wherein the glioma is a glioblastoma.

[0188] Embodiment 49. The method of any one of embodiments 46 to 48, wherein the tumor in the CNS is MTAP-deficient.

[0189] Embodiment 50. The method of any one of embodiments 46 to 49, wherein the tumor in the CNS is MT A- accumulating.

[0190] Embodiment 51. The method of any one of embodiments 43 to 50, wherein the PRMT5 inhibitor is administered systemically.

[0191] Embodiment 52. The method of any one of embodiments 43 to 51, wherein the PRMT5 inhibitor is administered orally.

[0192] Embodiment 53. The method of any one of embodiments 43-52. wherein the PRMT5 inhibitor is administered twice daily (BID).

[0193] Embodiment 54. The method of any one of embodiments 43 to 53, w herein the CDK4 / 6 inhibitor is administered systemically.

[0194] Embodiment 55. The method of any one of embodiments 43 to 54, w herein the CDK 4 / 6 inhibitor is administered orally.

[0195] Embodiment 56. Use of a protein arginine methyltransferase 5 (PRMT5) inhibitor in the manufacture of a medicament for treating a tumor in the central nervous system (CNS) in a subject in need thereof, wherein the PRMT5 inhibitor is a compound having the structural formula:IPTS'200237393.1Attorney Docket No TGO-037WOCompound 1;wherein the compound is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0196] Embodiment 57. The use of embodiment 56, wherein the tumor in the CNS is a glioma.

[0197] Embodiment 58. The use of embodiment 57, wherein the glioma is a glioblastoma.

[0198] Embodiment 59. The use of any one of embodiments 56 to 58, wherein the tumor in the CNS is MTAP-deficient.

[0199] Embodiment 60. The use of any one of embodiments 56 to 59. wherein the tumor m the C S is MTA-accumulating.

[0200] Embodiment 61. Use of a protein arginine methyltransferase 5 (PRMT5) inhibitor in the manufacture of a medicament for treating a cancer in a subject in need thereof, wherein the PRMI'5 inhibitor is a compound having the structural formula:F Compound 1;wherein the compound is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof,

[0201] Embodiment 62. The use of embodiment 61, wherein the cancer comprises a solid tumor with MTAP-deficiency.

[0202] Embodiment 63. The use of embodiment 61 or 62, wherein the cancer comprises glioma (e.g, glioblastoma), non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma.

[0203] Embodiment 64. The use of any one of embodiments 56 to 63, wherein the medicament is configured for systemic administration.IPTS / 200237393 1Attorney Docket No TGO-037WO

[0204] Embodiment 65. The use of any one of embodiments 56 to 64, wherein the medicament is configured for oral administration.

[0205] Embodiment 66. The use of any one of embodiments 56 to 65, wherein the medicament is configured for twice daily (BID) administration.[0206 [ Embodiment 67. The use of any one of embodiments 56 to 66, wherein the subject also receives (or wherein the medicament is configured for administration with) a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor having the structural formula:Compound 2:

[0207] wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0208] Embodiment 68. The use of embodiment 67, wherein the CDK 4 / 6 inhibitor is administered systemically.

[0209] Embodiment 69. The use of embodiment 67 or 68, wherein the CDK 4 / 6 inhibitor is administered orally.

[0210] Embodiment 70. Use of a protein arginine methyltransferase 5 (PRMT5) inhibitor in combination with a cyclin-dependent kinase (CDK) 4 / 6 inhibitor in the manufacture of a medicament for treating a cancer in a subject in need thereof, wherein:the PRMT5 inhibitor is a compound having the structural formula:H2N. X / NHUlf,- H 1uF-4-FF Compound 1;wherein Compound 1 is present m the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof andthe CDK4 / 6 inhibitor is a compound having the structural formula:IPTS'200237393.1Attorney Docket No TGO-037WOCompoundwherein Compound 2 is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0211] Embodiment 71. Use of a protein arginine methyltransferase 5 (PRMT5) inhibitor and a cychn-dependent kinase (CDK) 4 / 6 inhibitor in the manufacture of medicaments for treating a cancer in a subject in need thereof in combination wherein:the PRMT5 inhibitor is a compound having the structural formula:F Compound 1;wherein Compound 1 is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof; andthe CDK4 / 6 inhibitor is a compound having the structural formula:Compound 2:wherein Compound 2 is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof and wherein the medicaments can be administered concomitantly or sequentially.

[0212] Embodiment 72. The use of embodiment 70 or 71, wherein the cancer comprises a solid tumor with MTAP-deficiency.

[0213] Embodiment 73. The use of any one of embodiments 70-72, wherein the cancer comprises glioma (e.g., glioblastoma), non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcmoma.

[0214] Embodiment 74. Use of a protein arginine methyltransferase 5 (PRMT5) inhibitor in combination with a cyclin-dependent kinase (CDK) 4 / 6 inhibitor in the manufacture of a49IPTS'200237393.1Attorney Docket No TGO-037WOmedicament for treating a tumor in the central nervous system (CNS) in a subject in need thereof, wherein:the PRMT5 inhibitor is a compound having the structural formula:F Compound 1;wherein Compound 1 is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof; andthe CDK4 / 6 inhibitor is a compound having the structural formula:'F Compound 2;wherein Compound 2 is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof.

[0215] Embodiment 75. Use of a protein arginine methyltransferase 5 (PRMT5) inhibitor and a cyclin-dependent kinase (CDK) 4 / 6 inhibitor in the manufacture of medicaments for treating a tumor in the central nervous system (CNS) in a subject in need thereof in combination wherein:

[0216] the PRMT5 inhibitor is a compound having the structural formula:F Compound 1;wherein Compound 1 is present m the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof; andthe CDK4 / 6 inhibitor is a compound having the structural formula:IPTS'200237393.1Attorney Docket No TGO-037WOCompoundwherein Compound 2 is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof and wherein the medicaments can be administered concomitantly or sequentially.[02171 Embodiment 76. The use of embodiment 74 or 75, wherein the tumor in the CNS is a glioma.

[0218] Embodiment 77. The use of embodiment 76, wherein the glioma is a glioblastoma.

[0219] Embodiment 78. The use of any one of embodiments 74 to 77, wherein the tumor in the CNS is MT AP-defi cient.

[0220] Embodiment 79. The use of any one of embodiments 74-78, wherein the tumor in the CN S is MTA-accumulating.

[0221] Embodiment 80. The use of any one of embodiments 70 to 79, wherein the PRMT5 inhibitor medicament is configured for systemic administration.

[0222] Embodiment 81. The use of any one of embodiments 70 to 80, wherein the PRMT5 inhibitor medicament is configured for oral administration.

[0223] Embodiment 82. The use of any one of embodiments 70 to 81, wherein the PRMT5 inhibitor medicament is configured for twice daily administration.

[0224] Embodiment 83. The use of any one of embodiments 70 to 82, wherein the CDK.4 / 6 inhibitor medicament is configured for systemic administration.

[0225] Embodiment 84. The use of any one of embodiments 70 to 83, wherein the CDK4 / 6 inhibitor medicament is configured for oral administration.EXAMPLES

[0226] The following Examples are merely illustrative and are not intended to limit the scope or content of the invention in any way.IPTS'200237393.1Attorney Docket No TGO-037WOExample 1: Synthesis of (R)-N1-(4-amino-1H-pyrazolo[4,3-c]pyridin-7-yl)-N2-methyl-N2-(1-(4-(trifluoromethyl)phenyl)ethyl)oxalamide (Compound 1)

[0227] (R)-N1-(4-amino-1H-pyrazolo[4,3-c]pyridin-7-yl)-N2-methyl-N2-(1-(4- (trifluoromethyl)phenyl)ethyl)oxalamide can be prepared following the procedure outlined in the below scheme.

[0228] Step 1: 2,2,2-trifluoroethyl 2-[methyl-]l-[4- (trifluoromethyl)phenyl]ethyl|amino]-2-oxo-acetate

[0229] N-methyl-1-[4-(trifluoromethyl)phenyl]ethanamine (502.3 mg, 2.47 mmol) and Triethylamine (275.15 mg, 2.72 mmol, 378.99 pL) were dissolved in dichloromethane (DCM) (12 mL) and the resulting solution was cooled to -5°C in an ice / methanol bath. 2,2,2-trifluoroethyl 2-chloro-2-oxo-acetate (494.46 mg, 2.60 mmol) was added dropwise and the 52IPTS'200237393.1Attorney Docket No TGO-037WOresulting mixture was stirred overnight. Water (15 mL) was added to the reaction mixture and an organic layer was separated. The aqueous layer was extracted with DCM (15 mL) and combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain 2,2,2-trifluoroethyl 2-[methyl-[1-[4-(trifluoromethyl)phenyl]ethyl]amino]-2-oxo-acetate (811 mg, 2.27 mmol, 91.84% yield) as a light-yellow oil.

[0230] Liquid chromatography mass spectrometry (electrospray ionization) (LCMS(ESI)):[M+H]+m / z: calcd 358.09; found 358.2; Rt = 1.344 min.

[0231] Step 2: N'-methyl-N'-[1-[4-(trifluoromethyl)phenyl]ethyl]oxamide

[0232] 2,2,2-trifluoroethyl 2-[methyl-[1-[4-(trifluoromethyl)phenyl]ethyl]amino]-2-oxo-acetate (811 mg, 2.27 mmol) was dissolved in MeOH (10 mL) and NH3 / MeOH (15 mL) was added thereto. The resulting solution was stirred overnight. The reaction mixture was concentrated in vacuo to obtain N'-methyl-N'-[1-[4-(trifluoromethyl)phenyl]ethyl]oxamide (628 mg, crude).

[0233] LCMS(ESI): [M-H]’ m / z: calcd 273.08; found 273 0; Rt = 1.224 min.

[0234] Step 3: N-(4-amino-2-tetrahydropyran-2-yl-pyrazolo[4 -c]pyridin-7-yI)-N’-methyI-N'-[l-[4-(trifluoroniethyl)phenyl]ethyl]oxamide

[0235] All components were partitioned between two 8 ml vials. N'-methyl-N'-[1-[4-(trifluoromethyl)phenyl]ethyl]oxamide (300 mg, 1.09 mmol), 7-bromo-2-tetrahydropyran-2-yl-pyrazolo[4,3-c]pyridin-4-amine (357.57 mg, 1.20 mmol), Copper (3.48 mg, 54.70 μmol), Copper(I) iodide (104.17 mg, 546.97 μmol, 18.54 μL), rac-(1R,2R)-N1,N2-dimethylcyclohexane-1,2-diamine (116.70 mg, 820.45 pmol) and Cesium carbonate (712,85 mg, 2.19 mmol) were mixed in Dioxane (8 mL). The resulting mixture was sparged with argon for 5 min The vials were sealed and heated at 100°C over weekend. The reaction mixture was cooled and filtered. The filter cake was washed with MeOH (10 mL) and the filtrate was concentrated in vacuo to obtain N-(4-amino-2-tetrahydropyran-2-yl-pyrazolo[4,3-c]pyridin-7-yl)-N'-methyl-N'-[l-[4-(trifluoromethyl)phenyl]ethyl]oxamide (1 g, crude) as a greenish solid which was used in the next step without purification.

[0236] LCMS(ESI): [M+H]+m / z: calcd 491.2; found 491.2; Rt = 1.167 min.

[0237] Step 4: The synthesis of N-(4-amino-1H-pyrazolo[4,3-c]pyridin-7-yl)-N'-methyl-N'-[1-[4-(trifluoromethyl)phenyl]ethyl]oxamide

[0238] N-(4-amino-2-tetrahydropyran-2-yl-pyrazolo[4,3-c]pyridin-7-yl)-N'-methyl-N'-[1-[4-(trifluoromethyl)phenyl]ethyl]oxamide (1 g, 2.04 mmol) was dissolved in MeOH (5 mL) and HCl / Dioxane (5 mL) w as added thereto. The resulting mixture was stirred for 3 hr and 53IPTS- 200237393 1Attorney Docket No TGO-037WOthen concentrated in vacuo. The residue was purified by HPLC (0-2-10 min, 8-15-55 H2O / MeOH / 0.1NH4OH, flow 30 mL / min (loading pump 4 mL MeOH), target mass 407, column: XBridge BEH CIS 100*19mm, 5 microM) to obtain N-(4-amino-1H-pyrazolo[4,3-c]pyridin-7-yl)-N'-methyl-N'-[1-[4-(trifluoromethyl)phenyl]ethyl]oxamide (43.3 mg, 106.56 pmol, 5.23% yield) as a light-yellow solid.

[0239] 1H NMR (600 MHz, dmso) 8 1.29 - 1.67 (m, 3H), 2.57 - 2.92 (m, 3H), 5.36 - 5.87 (m, 1H), 6.61 - 6.91 (m, 2H), 7.49 - 7.59 (m, 1H), 7.61 - 7.65 (m, 1H), 7.65 - 7.71 (m, 1H), 7.72 - 7.78 (m. 2H), 8.13 - 8.21 (m, 1H), 9.65 - 10.73 (m, 1H), 12.47 - 13.25 (m, 1H).

[0240] LCMS(ESI): [M+H]+m / z: calcd 407.16; found 407.2; Rt = 1.010 min.

[0241] Step 5: Synthesis of (R)-N1-(4-amino-1H-pyrazolo[4,3-c]pyridin-7-yl)-N2-methyl-N2-(1-(4-(trifluoromethyl)phenyl)ethyl)oxalamide (Compound 1)

[0242] N-(4-amino-1H-pyrazolo[4,3-c]pyridin-7-yl)-N'-methyl-N'-[1-[4- (trifluoromethyl)phenyl ] ethyl] oxamide (36.9 mg, 90.81 pmol) was chirally separated (Column: Chiralcel OJ-H (250x20 mm, 5 mkm); Mobile phase: Hexane(0.1 %DEA)-IPA-MeOH, 80-10-10 Flow Rate: 12 mL / min; Column Temperature: 24°C; Wavelength: 205 nm; RetTime = 39.73 min) to obtain (R)-N1-(4-amino-1H-pyrazolo[4,3-c]pyridin-7-yl)-N2-methyl-N2-(1-(4-(trifluoromethyl)phenyl)ethyl)oxalamide (15.95 mg, 39.25 μmol, 86.45% yield) as a light-yellow solid.

[0243] ’H NMR (600 MHz, dmso) 8 1.25 - 1.71 (m, 3H), 2.74 (d, 3H), 5.36 - 5.84 (m, 1H), 6.60 - 6.85 (m, 2H), 6.89 - 7.68 (m, 3H), 7.69 - 7.78 (m, 2H), 8.12 - 8.22 (m, 1H), 9.65 - 10.67 (m, 1H), 12.62 - 13.36 (m, 1H).

[0244] LCMS(ESI): [M+H]+m / z: calcd 407.16; found 407.0; Rt = 2.269 min.

[0245] The absolute stereochemistry of Compound 1 was independently confirmed.Example 2: In Vitro Antiproliferative Activity of Compound 1 in a Multilineage Cell Line Panel

[0246] This example characterizes the selectivity of Compound 1 for MTAP-null cancer cell lines relative to MTAP wild type (WT) cancer cell lines. Compound 1 was profiled in a panel of 142 cell lines in a 7-day in vitro viability assay. The panel was comprised of cell lines representing bladder cancer, bone sarcoma, glioblastoma, breast cancer, esophageal cancer, hepatocellular carcinoma, leukemia, liver / bile duct cancer, lymphoma, mesothelioma (lung pleura), lung cancer, pancreatic cancer, skin cancer, and stomach cancer. The cell lines were propagated and assayed under the appropriate reported cell culture conditions. The cell 54IPTS- 200237393 1Attorney Docket No TGO-037WOlines were dosed with a titration of Compound 1. Cellular viability7was assessed using CellTiter-Glo following 7 -days of inhibitor treatment.

[0247] To visualize the selectivity of Compound 1 for MTAP-null cancer cell lines relative to MTAP WT cancer cell lines and to correct for differences in compound potency, a waterfall plot (FIG. 1) was generated using the maximum effect (Amax) of Compound 1 at a concentration equal to 10X the TC50of Compound 1 (20nM) in the MAPI MTAP-null cancer cell line in a 7-day viability assay. The data demonstrate that Compound 1 is highly' potent and selective (about 55 times selective) for MTAP-null cancer cell lines in this broad, multilineage cell line panel.Example 3: Evaluation of Antitumor Activity of Compound 1 as a Single Agent in MTAP-null Patient Derived Xenograft Models

[0248] This example tests antitumor activity of Compound 1 surveyed in a panel of MTAP-null patient-derived xenograft (PDX) models representing squamous non-small cell lung cancer (NSCLC), glioblastoma, pancreatic cancer, and cholangiocarcmoma in BALB / c nude immune-deficient mice,

[0249] V ehicle (5% DMA and 20% Captisol) or Compound 1 was dosed at 30 mg / kg BID or 90 mg / kg BID in BALB / c nude mice. Tumor-bearing mice (n = 3-6 / group) were assigned to treatment groups with average starting tumor volumes (TVs) ranging from 130 mm3to 250 mm3. Treatments of Compound 1 up to 90 mg / kg were generally tolerated in these studies.

[0250] Tumor volumes were determined in individual mice over the duration of the study for all models. The %T / C or %TV (when the final TV was smaller than the initial TV) values were plotted for all 4 PDX models as shown in FIGs.2A-2D. As shown in FIG.2A-2D, both 30 mg / kg BID and 90 mg / kg BID doses of Compound 1 showed antitumor responses in the tested PDX models.Example 4: Evaluation of Antitumor Activity of Compound 1 as a Single Agent in LU99 Human Lung Cancer Xenograft Model

[0251] This example tests antitumor activity of Compound 1 as a monotherapy in the MTAP-null, KRAS-mutant LU99 human NSCLC xenograft model. Vehicle (5% DMA and 20% Captisol) or Compound 1 was dosed at 10 mg / kg BID, 30 mg / kg BID, or 90 mg / kg BID in BALB / c nude mice. Tumor-bearing mice (n = 8 / group) were assigned to treatment groups with average starting tumor volumes (TVs) of ~216 mm3. The mean tumor volume of Vehicle control reached an average of 2522 mm3by Day 20.55IPTS- 200237393 1Attorney Docket No TGO-037WO

[0252] Tumor volumes were determined in individual mice over the duration of the study As shown in FIG.3, Compound 1 achieved significant antitumor activity at all doses tested, and 96% tumor growth inhibition and deep tumor regression at the 30 mg / kg BID and 90 mg / kg BID single agent doses, respectively.Example 5: Evaluation of Antitumor Activity of Compound 1 in Combination with Compound 2 in LU99 Human Lung Cancer Cell line and Xenograft Model

[0253] This example evaluates the in vitro and in vivo efficacy of Compound I in combination with Compound 2 (abemaciclib) in the LU99 MTAP-deleted non-small cell lung cancer (NSCLC) model.In Vitro Efficacy

[0254] The LU99 MTAP-deleted NSCLC cell line was propagated and assayed under the appropriate reported cell culture conditions. Two independent methodologies were used to determine an in vitro combination benefit: a 14-day CellTiter-Glo assay and a 12-day colony formation assay. For the CellTiter-Glo assay, the cell line was dosed with a 9x5 combination matrix of Compound I (TNG456) and abemaciclib. For the colony formation assay, the cell line was dosed with a 5x3 combination matrix of Compound 1 (TNG456) and abemaciclib and assayed using crystal violet staining.

[0255] In the CellTiter-Glo assay, all of the data were normalized to the dimethyl sulfoxide (DMSO)-treated control cells and fit using a non-linear 4-parameter variable slope algorithm (FIG.4A). For the colony formation assay, the crystal violet staining was quantified from wells treated with either a DMSO vehicle, single agent Compound 1 (TNG456) or abemaciclib, or combination of Compound 1 (TNG456) and abemaciclib. The concentrations of Compound 1 (' TNG 456) and abemaciclib used in this assay were chosen to be equally efficacious and causing -50% loss of viability (FIG.4B).

[0256] By determining viability with either a CellTiter-Glo assay or a colony formation assay, a combination benefit between Compound 1 (TNG456) and abemaciclib was observed in the LU99 MTAP-deleted NSCLC cell line. Further analyses of the CellTiter-Glo data to determine whether combining Compound 1 (TNG456) with abemaciclib is synergistic, additive or antagonistic strongly suggest that the combination is additive or synergistic depending on the dose These in vitro data support the clinical potential for a combination of Compound I (TNG456) and abemaciclib in treating MTAP-deleted cancers representing multiple tumor histologies such as NSCLC.56IPTS- 200237393 1Attorney Docket No TGO-037WOIn Vivo Efficacy

[0257] For the in vivo efficacy study, vehicle (5% DMA and 20% Captisol co-dosed with 1% hydroxyethyl cellulose + 0.1% antifoam in 25 mM PB pH 2), Compound 1 (TNG456) at the minimum efficacious dose of 30 mg / kg BID was dosed as single agent or combined with abemaciclib at QD doses of 50 mg / kg in LU99 tumor-bearing mice (Table 1 provides study design). The duration for the study was 28 days. As shown in FIG. 4C, all treatments were well-tolerated with < 5% body weight loss.Table 1: Study DesignT.. Animals / Dose Dose Vol „. „.c_ Treatment „,.,,,, Route RegnnencDuration Group (mg / kg) (pL / g)TNG456 Vehicle, ■3o- 10 p.o. BID, abemaciclib 8A21 days Vehicle5’10 p o'QDTNG456 _ 8 _ 30 _ 10 p.o. BID 28 days abemaciclib 8 50 10 p.o. QD 28 days abemaciclib ~ 50 10 p.o. QD3'SAbbreviations: BID, twice daily: p.o., oral; QD, once daily.a. TNG456 Vehicle: 5% DMA+20% Captisolb. Abemaciclib Vehicle: 1% hydroxyethyl cellulose + 0.1% antifoam in 25 mM PB pH 2 c. The BID treatment interval was 8:16 hours

[0258] In the study, starting tumor volumes were ~198 mm3(FIG. 4D). The mean tumor volume of Vehicle control reached an average of 2121 mm3by Day 19. Compound 1 (TNG456) at 30 mg / kg BID exhibited strong antitumor activity as a single agent, with mean tumor growth suppression achieving tumor stasis (%T / C of 9). Abemaciclib exhibited minimal antitumor activity' as a single agent, with limited tumor growth suppression (%T / C of 76, w'hich was not statistically significant compared to Vehicle) at the 50 mg / kg QD clinically relevant dose. When 30 mg / kg BID TNG456 and 50 mg / kg QD abemaciclib were combined, near tumor regression (%T / C of 0) was achieved, suggesting a combination benefit of TNG456 with abemaciclib.

[0259] The adjusted plasma free unbound exposures of TNG456 were 1,521 h*ng / ml and 1,692 h*ng / ml for the 30 mg / kg monotherapy and in combination with abemaciclib, respectively. The adjusted plasma free unbound exposure of 50 mg / kg QD abemaciclib was 1,090 h*ng / ml in combination with TNG456, which was 1.8-fold higher compared to monotherapy at the same dose (601 h*ng / ml).IPTS- 200237393 1Attorney Docket No TGO-037WO

[0260] In summary, these data demonstrate a significant combination benefit of Compound 1 (TNG456) and abemaciclib compared with Compound 1 (TNG456) alone, at well-tolerated and clinically relevant exposures.Example 6: Pharmacokinetic / Pharmacodynamic (PK / PD) Study And Evaluation of Antitumor Activity of Compound 1 in MTAP-null Glioblastoma Xenograft Model (U87MG)Pharmacokinetic / Pharmacodynamic (PK / PD)

[0261] A 7-day PK / PD study was carried out in the U87MG MTAP-null glioblastoma xenograft model w ith the primary aim of establishing the correlation between Compound I (TNG456) plasma levels and the inhibition of PRMT5 activity, measured by variations in the levels of a representative SDMA-modified protein. The dose levels of Compound 1 (TNG456). along with the timing for plasma PK and tumor PD data collection, were selected according to the results from a high-dose PK study, as well as outcomes from a tolerability study. Dose levels up to 180 mg / kg were well-tolerated in the 7-day tolerability study.

[0262] In the study, female BALB / c nude mice bearing U87MG tumors were assigned to treatment groups with similar mean tumor volumes of 326 mm3and were treated with either vehicle (5% DMA, 20% Captisol) or Compound 1 (TNG456) at doses of 3 mg / kg BID. 10 mg / kg BID, 30 mg / kg BID, 90 mg / kg BID, or 120 mg / kg BID (Table 2 provides the study design for this study). All groups received treatment for 7 days. Plasma samples were collected at 1, 2, 4, 8, 16, and 24 hours post last dose and analyzed for TNG456 concentrations. Tumor samples were harvested at 8 hours and 24 hours after the last dose and processed for the determination of SDMA-modified protein levels through western blot analyses. Neither body weight loss (defined as > 5%) nor adverse clinical signs were observed during this study.58IPTS-200237393 1Attorney Docket No TGO-037WOTable 2: Study DesignGrou _ Animals Dose Dose Vol Rout Regimen Duratio Drug ®ps / group (mg / kg) (pl / g) ean 1 Vehicle 8 - 10 p.o. BID 7 days 2 TNG45 8 3 10 p.o. BID 7 days 3 TNG45 8 10 10 p.o. BID 7 days 4 TNG45 8 30 10 p.o. BID 7 days 5 TNG45 8 90 10 p.o. BID 7 days 6 TNG45 8 120 10 p.o. BID 7 days Abbreviations: BID, twice daily; p.o., oral.a. The BID treatment interval was 8: 16 hours.

[0263] Plasma concentrations of Compound 1 (TNG456) were dose-dependent, with maximal concentrations observed at 1-2 hours post last dose (FIG. 5A). The adjusted plasma free unbound exposures, based on a 16.3% free fraction in BALB / c nude mouse plasma, were 92 h*ng / ml, 259 h*ng / ml, 1217 h*ng / ml, 6004 h*ng / ml, and 16260 h*ng / ml for the 3 mg / kg BID, 10 mg / kg BID, 30 mg / kg BID, 90 mg / kg BID, and 120 mg / kg BID doses, respectively. As shown in FIG. 5A, U87MG tumor SDMA-modified protein levels decreased in a dose-dependent manner, with a maximal inhibition of 91 % and 93% (compared to Vehicle) observed at 8 hours post last dose of 90 and 120 mg / kg BID, respectively, and 92% observed at 24 hours after the last dose of 90 mg / kg BID.

[0264] At the minimally efficacious dose (MED) of 30 mg / kg BID, as determined in the efficacy study detailed below, the adjusted free unbound plasma concentrations of Compound 1 (TNG456). corresponding to an exposure of 1217 h*ng / ml, were sufficient to maintain coverage of the HAP1 MT AP -null antiproliferative ICso for approximately 20 hours (FIG. SA). At 90 mg / kg BID. the adjusted free unbound plasma concentrations of Compound 1 (TNG456), corresponding to an exposure of 6004 h*ng / ml, were sufficient to maintain coverage of the HAP1 MTAP-null antiproliferative IC50for more than 24 hours. These data are consistent with the profound PD modulation of S DMA-modified protein (shown in FIG.5A). 120 mg / kg BID, the adjusted free unbound plasma concentrations of Compound 1 (TNG456) were above the antiproliferative IC50coverage for MTAP WT HAP1 cells for almost 24 hours, which is consistent with the adverse clinical signs in the efficacy study. Antitumor Activity

[0265] The antitumor activity of Compound 1 (TNG456) was evaluated in the U87MG tumor xenograft model. Vehicle (5% DMA and 20% Captisol) or Compound 1 (TNG456) were dosed orally at 10 mg / kg BID, 30 mg / kg BID, or 90 mg / kg BID for 25 days. Prior toIPrS- 200237393 1Attorney Docket No TGO-037WOstudy enrollment, female BALB / c nude mice bearing U87MG tumors were randomly assigned to treatment groups (n = 8 / group) with similar starting mean tumor volumes of 159 mm3. Treatments of 10 mg / kg BID, 30 mg / kg BID, or 90 mg / kg BID were tolerated with an average body weight loss of <10% (FIG. 5B)Table 3: Study DesignGrou „ Animals Dose Dose Vol Rout Regimen Duratio Drugp / group (mg / kg) (pl / g) ean 1 Vehicle 8 - 10 p.o. BID 25 days 2 TNG458 10 10 p oBID 25 days TNG453 g 8 30 10 p.o. BID 25 days TNG454 g 8 90 10 p.o. BID 25 days Abbreviations: BID, twice daily; p.o., oral.a. The BID treatment interval was 8:16 hours.

[0266] Efficacy results are summarized in Table 4 and the corresponding longitudinal TV curves are depicted in FIG, 5C Tumors in the vehicle control group (Group 1) grew robustly, reaching a mean TV of 1558 mm3after 25 days of treatment. Treatment with Compound 1 (TNG456) resulted in dose-dependent antitumor activity, with %T / C values of 50 and 16 for the 10 mg / kg BID and 30 mg / kg BID treatment groups, respectively. Mean tumor regression of 57% was observed in the 90 mg / kg BID treatment group. The minimally efficacious dose (MED) of Compound 1 (TNG456) in the U87MG model was determined to be 30 mg / kg BID or less, as this dose and schedule resulted in near tumor stasis (%T / C of 16).IPTS / 200237393 1Attorney Docket No TGO-037WOTable 4: Antitumor Activity of Compound 1 (TNG456) as a Single Agent in the Treatment of U87MG Subcutaneous Xenograft ModelTV3(mm3) T / C TV „.P Valuediuli ( / «) ( / o)P PCG- nDo0 P PCG- DD2233b cG1 G2 Gvs3 GS41158±6 1558 3862 159±20 856±95 50 - ns 3 159±18 390±81 16 - ** ** 4 159±20 69+5 - -57 **** * ***Abbreviations: ns, not significant, PG, post-grouping; T / C, treatment / control; TV, tumor volume; vs, versus.a. Mean i SEM post treatment initiationb. %T / C= (Treated TVfinai-Treated TVinltiai) / (Vehicle TVfinai-Vehicle TVinitiai)* 100;c. %TV= [mean TVfinai - mean TVinitiai] *100d. Mixed-effect analysis followed by Fisher’s LSD test; * <0.05, ** <0.01, *** <0.001, **** P<0.0001, ns, not significant

[0267] In summary; the PK / PD relationship and antitumor activity of Compound 1 (TNG456) was dose-dependent Maximum PD modulation following 7 days of Compound I (TNG456) treatment was observed at 90 mg / kg BID and 120 mg / kg BID, with effects noted at 8 hours post last dose and remaining significantly suppressed up to 24 hours post last dose.Example 7: Evaluation of Antitumor Activity of Compound 1 in Combination with Compound 2 in MTAP-null Glioblastoma Xenograft Model (U87MG)

[0268] This example evaluates the antitumor activity of Compound 1 (TNG456) in combination with Compound 2 (abemaciclib) in the U87MG MTAP-null human glioblastoma model.In Vitro Efficacy

[0269] The MTAP-null human glioblastoma xenograft model (U87MG) cell line was propagated and assayed under the appropriate reported cell culture conditions. Two independent methodologies were used to determine an in vitro combination benefit: a 14-day CellTiter-Glo assay and a 12-day colony formation assay For the CellTiter-Glo assay, the cell line was dosed with a 9x5 combination matrix of Compound 1 (TNG456) and abemaciclib. For the colony formation assay, the cell line was dosed with a 5x3 combination matrix of Compound 1 (TNG456) and abemaciclib and assayed using crystal violet staining.61IPTS- 200237393 1Attorney Docket No TGO-037WO

[0270] In the CellTiter-Glo assay, all of the data were normalized to the dimethyl sulfoxide (DMSO)-treated control cells and fit using anon-linear 4-parameter variable slope algorithm (FIG. 6A). For the colony formation assay, the crystal violet staining was quantified from wells treated with either a DMSO vehicle, single agent Compound 1 (TNG456) or abemaciclib, or combination of Compound 1 (TNG456) and abemaciclib. The concentrations of Compound 1 (TNG456) and abemaciclib used in this assay were chosen to be equally efficacious and causing -50% loss of viability (FIG. 6B).

[0271] By determining viability' with either a CellTiter-Glo assay or a colony formation assay, a combination benefit between Compound 1 (TNG456) and abemaciclib was observed in the MTAP-null human glioblastoma xenograft mode] (U87MG) cell line. Further analyses of the CellTiter-Glo data to determine whether combining Compound 1 (TNG456) with abemaciclib is synergistic, additive or antagonistic strongly suggest that the combination is additive or synergistic depending on the dose. These in vitro data support the clinical potential for a combination of Compound 1 (TNG456) and abemaciclib in treating MT P-deleted cancers representing multiple tumor histologies such as glioblastoma.In Vivo Efficacy

[0272] For the in vivo efficacy study, vehicle (5% DMA and 20% Captisol co-dosed with 1% hydroxy ethyl cellulose + 0.1% antifoam in 25 mM PB pH 2). Compound 1 (TNG456) at the minimum efficacious dose of 30 mg / kg BID was dosed as single agent or combined with abemaciclib at 50 mg / kg QD in U87MG tumor-bearing mice (Table 5 provides study¬ design). Dosing was initiated in tumor-bearing mice when mean tumor volume reached 165 mm3. The mice w ere dosed until humane endpoint (tumor volume equal to -2000 mm3) was achieved (up to 87 days)Attorney Docket No TGO-037WOTable 5: Study Design„ Animals / Dose Dose Vol „rMaximum Treatment „.....,., Route Regimenc,Group (mg / kg) (pL / g) ® duration TNG456 Vehicle3_abemaciclib 8 P'0' „„ 21 daysVehicle11"10 QDTNG456 _ 8 _ 30 _ 10 p.o, BID 85 days abemaciclib 8 50 10 p.o. QD 70 days ~3{!-- - o7T - abemaciclib50 10 p.o. QDa^SAbbreviations: BID, twice daily; p.o, oral; QD, once daily,a. TNG456 Vehicle: 5% DMA+20% Captisolb. Abemaciclib V ehicle: 1 % hydroxyethyl celldose + 0 1 % antifoam in 25 M PB pH 2 c. The BID treatment interval was 8:16 hours

[0273] FIG.6C is a Kaplan meier survival curve in an MTAP-null GBM (U87MG) orthotopic CDX model treated with 30 mg / kg BID Compound 1 (TNG456 in FIG. 6C) as a single agent or m combination with abemaciclib dosed at 50 mg / kg QD. * refers to p-value <0.05 and ** refers to p-value <0.0.1 according to log-rank analysis (Mantel-Cox test) As shown in FIG.6C, strong antitumor activity was demonstrated in the orthotopic model and supports a significant combination benefit of Compound 1 (TNG456) and abemaciclib compared with either Compound 1 (TNG456) or Compound 2 (abemaciclib) alone, at well- tolerated and clinically relevant exposures.Example 8: Evaluation of Antitumor Activity of Compound 1 in Combination with Compound 2 in MTAP-null Human Glioblastoma Xenograft Model (AM38)

[0274] This example evaluates the antitumor activity7of Compound 1 (TNG456) in combination with Compound 2 (abemaciclib) in the AM38 MTAP-null human glioblastoma xenograft model.

[0275] Vehicle (5% DMA and 20% Captisol co-dosed with 1% hydroxyethyl cellulose + 0.1% antifoam in 25 mM PB, pH 2), 45 mg / kg BID Compound 1 (TNG456) single-agent or combined with 50 mg / kg QD Compound 2 (abemaciclib), or 90 mg / kg BID Compound 1 (TNG456) single-agent or combined with 20 mg / kg QD Compound 2 (abemaciclib) were dosed for up to 15 days, except for 90 mg / kg BID TNG456 single-agent and the combination groups, which had extended treatment periods as outlined in Table 6. As shown in FIGs. 7 A and 7C. all treatments were generally tolerated with an average body weight loss of <10%.63IPTS-200237393 1Attorney Docket No TGO-037WOTable 6: Study Design_ Animals / Dose Dose Vol „c„ Drug „.,,..,,. Route RegimencDuration Group (mg / kg) (pL / g)TNG456 Vehicle3abemaciclib 8 'p°'r15 days Vehicle” ■5 P°-QDTNG456 8 45 10 p o. BID 15 days TNG456 8 90 10 p.o. BID 41 days abemaciclib 8 20 10 p.o. QD 15 days abemaciclib 8 50 10 p.o. QD 15 days TNG456 45 10 p.o. BID abemaciclib 50 10 p.o. QDC a^STNG456 90 10 p.o. BID, abemaciclib ' 20 10 p o. QD1 a'SAbbreviations: BID, twice daily; p.o,, oral; QD, once daily.a. TNG456 Vehicle: 5% DMA+20% Captisolb. Abemaciclib Vehicle: 1% hydroxy ethyl cellulose + 0.1% antifoam in 25 mM PB pH 2 c. The BID treatment interval was 8:16 hours

[0276] Starting tumor volumes were -206 mm3. The mean tumor volume of the Vehicle control group reached an average of 1969 mm-1by Day 13. Abemaciclib did not exhibit single-agent antitumor activity, either at the sub-therapeutic dose of 20 mg / kg QD or at the clinically relevant dose of 50 mg / kg QD (%T / C of 107 and 85, respectively). TNG456 showed minimal antitumor activity' at 45 mg / kg BID (%T / C of 71) but demonstrated moderate tumor growth suppression at 90 mg / kg BID (%T / C of 42) (Table 7 summarizes results from the efficacy study).

[0277] In contrast, strong tumor growth inhibition (%T / C of 27 and 33, respectively) was achieved in the combination groups of TNG45645 mg / kg BID with abemaciclib 50 mg / kg QD, as well as TNG456 90 mg / kg BID with abemaciclib 20 mg / kg QD, by the time the Vehicle group reached the humane endpoint on Day 13 (Table 7). Dosing for both combination groups continued for an additional 21 days of treatment until the interim report was drafted and is still ongoing, with the mean tumor volumes continuing to decrease, suggesting a strong synergistic effect of TNG456 and abemaciclib in the AM38 glioblastoma model (FIGs. 7B and 7D).IPTS-200237393 1Attorney Docket No TGO-037WOTable 7: Antitumor Activity of TNG456 in Combination with Abemaciclib in AM38 Subcutaneous TumorsTVa(mm3) PVaIuec dTreatment, vs mg / kgpG pG pG D13 vs TNG456Abemacic DO D13 D46 (%)bVehicle at same,lb at, same dose.dose TNG456Vehicle 2064 196943abemaciclib 20 72VehicleTNG45645 2064 145842BID 21 09 "1 RSabemaciclib 50 2074 170642o,,,cQD 19 05 "85TNG45645BID 2074 678411 280 18 ***<.abemaciclib 50 20 9 4QD TNG45690 2074 940415 42BID 19 5abemaciclib 20 2074 209143QD 21 51 ’107 nsTNG45690BID 2074 793414 457 13abemaciclib 20 23 9 4QDAbbreviations: BID, twice daily; ns, not significant; PG, post-grouping; QD, once daily; T / C, treatment / control; TV, tumor volume; vs, versusa. Group Mean SEM post treatment initiationb. %T / C= (Treated TVfmai-Treated TVinitiaj) / (Vehicle TVfmai- Vehicle TVinitiaj)*100; calculated at DI 3c Mixed-effect analysis on D13 followed by Fisher's LSD test; * <0 05, ** P<0 01, *** P<0.001, **** P<0.0001, ns: not significant.d. Mixed-effect analysis on D34 followed by Fisher’s LSD test; * P<0,05, ** P<0.01, *** P<0.001. **** P<0.0001, ns: not significant.Example 9: Evaluation of the Antitumor Activity of Compound 1 as a Single Agent in ST458 Bladder Carcinoma Patient-Derived Xenograft Model

[0278] The antitumor activity of Compound 1 as a single agent was further evaluated in the MTAP-null PDX model ST458 (bladder carcinoma). Vehicle control (5% DMA and 20%Attorney Docket No TGO-037WOCaptisol) or Compound 1 were dosed at 30, 60, or 90 mg / kg BID for up to 51 days, as outlined in Table 8. Prior to study enrollment, tumor-bearing female BALB / c nude mice were randomly assigned into treatment groups (n = 6 / group) with similar starting mean TVs of 220 mmJ. All treatments for both studies were generally tolerated with an average body weight loss of <10% (FIG. 8A).Table 8: Study DesignST458. Dose _, „, „. _„ Animals, „ Dose Vol Rout Regimen Duratio Grou Drug, (mg / kg.,,p / group (pl / g) eaan 1 Vehicle 6 - 10 p.o BID 20 days2 6 30 10 p.o. BID 51 davsd 13 Compoun6 60 10 B1£) 5]d 14 A 6 90 10 p.o. BID 51 davsd 1Abbreviations: BID, twice daily; p.o., oral.a. The BID treatment interval was 8:16 hours.

[0279] The mean tumor volume of G1 (Vehicle control) reached an average of 2028 mm3by Day 20, leading to the euthanasia of this group due to tumor burden (Table 9) Compound 1 exhibited strong antitumor activity, with a mean 41%, 47% and 52% tumor regression for the 30, 60 and 90 mg / kg BID groups, respectively, at Day 20 (Table 9). By the end of the study on Day 51. 65%. 75% and 87% tumor regressions were noted for the 30, 60 and 90 mg / kg BID groups, respectively (FIG. 8B). The adjusted plasma free unbound concentrations of Compound 1 were dose-dependent. The plasma exposures were 1497, 2837 and 5802 h*ng / ml for the 30, 60 and 90 mg / kg BID doses, respectively.IPTS / 200237393 1Attorney Docket No TGO-037WOTable 9: Antitumor Activity7of Compound 1 as a Single Agent in the Treatment of ST458 Subcutaneous PDX TumorsTV’Cnim3) TV- TV- P ValuedGiou D20 D51P F P rG-D u0uPDG20’DPG51" (%b)7(%c) GVS1 GVS2 GVS3 GVS4 1 220±172°O5±32 220 17 13029 78±30 -41 -65 **** _ _ _ 3 222±20 118±19 55±18 -47 -75 **** ns4 223±16 106±16 29±11 -52 -87 **** ns ns Abbreviations: ns, not significant; PDX, patient-derived xenograft; PG, post-grouping; TV. tumor volume; vs, versus.Note: a. Group Mean ± SEM post treatment initiationb. %TV = [mean TVfmai- mean TVinitiai / TVinitiai]*100, calculated at D20c. %TV = [mean TVfmai - mean TVmiiiai / TVmit.ai]*100, calculated at D51d. Mixed-effect analysis followed by Fisher’s LSD test; * P<0.05, ** P<0.01, ***P<0.001, **** P<0.0001, ns: not significant.

[0280] Together, these data demonstrate that in the ST458 MTAP-nuIl PDX xenograft model Compound 1 drives deep and sustained tumor regressions at well-tolerated exposures.Example 10: Determination of brain exposure for Compound 1 in male and female beagle dogs

[0281] The purpose of this study was to determine the brain exposure in the male and female beagle dogs following administrations of TNG456 to male and female beagle dogs once-daily7by oral gavage for 14 consecutive days.

[0282] Sixteen (8 males and 8 females) beagle dogs were divided into four groups with 2 animals / sex / group. Animals in Groups 1 to 4 were administered TNG456 by once-daily oral administration at 0 (vehicle control), 5, 15 and 60 mg / kg in 0.5% (w / v) MC in water for 14 consecutive days, respectively.

[0283] Plasma and brain samples were collected at necropsy after 14-day dosing for Groups 1 to 3 and after 9 day dosing for Group 4. Concentrations of TNG456 in plasma and brain samples were determined by a liquid chromatography tandem mass spectrometry (LCMS / MS) method The Kp,uu ranged from 0607 - 1 071 for 5 mg / kg dosing group, from 0.459 - 0.667 for 15 mg / kg dosing group and from 0.478 - 0.592 for 60 mg / kg dosing group.67IPrS- 200237393 1Attorney Docket No TGO-037WOTable 10. Plasma and brain concentrations of TNG456 following repeated oral administration at 5. 15 and 60 mg / kgBrain PlasmaDoseStudy Concent Coneent Average Average (mg / kg / SexDay ration ration KpaKp Kp,uu^ Kp,uu d ay)(ng / g) (ng / ml)296 76.9 3.85 0.920Male569 168 3.39 0.8095 15 3.66 0.876162 60.3 2.69 0.642 Female189 39.9 4.74 1.1321370 464 2.95 0706Male1440 709 2.03 0.48515 15 2.36 0.5631530 738 2.07 0.495 Female1010 426 2.37 0.5672830 987 2.87 0.685Male2220 1050 2.11 0.50560 10 2.53 0.6053720 1420 2.62 0.626Female2030 805 2.52 0.603‘ BQL’' means below the quantifiable limit. ” means not calculated.3Brain-to-plasma partition coefficient (Kp) = Concentration brain / Concentration plasmabUnbound brain-to-plasma partition coefficient (Kp,uu) = Concentration brain, unbound / Concentration plasma, unbound (unbound % mean dog plasma= 1046%, unbound dog bram=2.5%)Example 11: Determination of brain exposure for Compound 1 in male cynomolgus monkeys

[0284] The aim of this study was to assess the brain exposure in the male cynomolgus monkeys following tw o oral administrations of Compound 1, given 8 hours apart

[0285] Three male cynomolgus monkeys with surgically placed cistema magna ports received two oral doses of Compound 1, each at 10 mg / kg in 0.5% (w / v) MC in water. These ports enable direct serial sampling of cerebrospinal fluid (CSF). which serves as a surrogate for free brain concentration.

[0286] Plasma and CSF samples at various time points w^ere collected for up to 24 hours after the first dose. Concentrations of Compound 1 in plasma and CSF samples were 68IPTS' 200237393 ]Attorney Docket No TGO-037WOdetermined by a liquid chromatography tandem mass spectrometry (LCMS / MS) method. The Kp.uu was calculated as follows:AUC0-24h, CSF / [AUC0-24h, plasma * fu, monkey plasma] where:• AUC0-24h. CSF: area under the CSF concentration-time curve over the last 24 hours.• AUC0-24h, plasma: area under the plasma concentration-time curve over the last 24 hours.® fu, monkey plasma: the fraction of TNG456 that is not bound to monkey plasma proteins

[0287] The kp,uu values ranged from 0.45 to 0.90 The plasma and CSF concentrations as a function of time are shown in FIG. 10.Example 12: Additional pharmacokinetic studies of Compound 1

[0288] The PK properties of Compound 1 were evaluated in beagle dogs and cynomolgus monkeys (Table 11). Following a 1 mg / kg IV dose in beagle dogs (n = 3), clearance was 3.0 mL / min / kg, volume of distribution w as 1.5 L / kg, and the terminal half-life was 6.4 h. Oral administration of a 3 mg / kg dose to beagle dogs resulted in a Cmax of 0.802 pg / mL. AUCinf of 9.2 h-ug / mL, with 55 % bioavailability. In cynomolgus monkeys (n - 3), a 1 mg / kg IV dose of Compound 1 gave a clearance of 16 mL / min / kg, volume of distribution of 4.0 L / kg, and half-life of 3.9 h. Oral administration at 3 mg / kg produced a Cmax of 0.366 pg / mL, AUCmf of 2.55 h-ug / mL, and 85 % bioavailability'. The predicted human half-life of Compound 1 is approximately 6 hours, suggesting that a twice-daily (BID) dosing regimen would maintain exposures with a Cmax / Cmin ratio < 2. This regimen is projected to sustain concentrations above the MT AP -null Gbo at trough while keeping peak exposures below those expected to affect MTAP WT cells. Brain penetration of Compound 1 was confirmed in two independent cynomolgus monkey CSF studies (including the study of Example 11), which demonstrated mean KP,uu values of 9.59 (0.57 and 0 1 in individual studies). Consistent CNS exposures were also observed in beagle dogs, with brain and time-matched plasma samples collected at the end of anon-GLP dose-range finding toxicity study with KP,uu values averaging 0.64 (ranging from 0.44 to 1.1 ).Attorney Docket No TGO-037WOTable 11. in vivo PK Characterization of Compound IClearanceSpecies Vdss(L / kg) T: 2 (h) %F PPBCKp,uu (niL / min / kg)3.0 1.5 64 55 10 064 DogaCynomolgus16 4.0 3.9 85 19 0.59 Monkeyb

[0289] aIV / PO dosing in beagle dog (vehicle. IV: 1 mg / kg solution of 1% v / v DMSO / 99% of 20% w / v HP-P-CD in saline; PO: 3 mg / kg suspension in 0.5% MC, n = 3 per arm),bIV / PO dosing in cynomolgus monkey (vehicle, IV: 1 mg / kg solution of 1% v / v DMSO / 99% of 20% w / v HP-p-CD in saline; PO: 3 mg / kg suspension in 0.5% MC, n = 3 per arm).cPlasma Protein Binding, % unbound.Example 13: Clinical Study of Compound 1

[0290] Preclinical experimental data has shown that Compound 1 has a half-maximal inhibitory concentration (IC50) of 2 nM in a pharmacodynamic (PD) study and a growth inhibition 50 (GIso) of 20nM. Compound 1 exhibits about 55 times selectivity in MTAP-deleted cancers relative to cancers without MTAP deletion.

[0291] Unbound brain-to-plasma partition coefficient (Kp.uu) is a crucial parameter in understanding brain penetrance for drugs targeting the central nervous system (CNS). It indicates how effectively a drug can cross the blood-brain barrier (BBB) and reach its site of action in the brain. Studies have shown that Compound 1 is bram-penetrant and exhibits a preclinical Kp,uu range of 0.5-1.1 as measured in non-human primates and dogs.

[0292] Clinical activity of Compound 1 (TNG456) is being evaluated in solid tumors with MTAP deletion, including advanced or metastatic solid tumors with CNS involvement such as glioblastoma, in a Phase I / II clinical study designed as shown m FIG. 9. The study evaluates Compound 1 as a single agent and m combination with abemaciclib in patients with MTAP-deleted cancers.INCORPORATION BY REFERENCE

[0293] The entire disclosure of each of the patent documents and scientific articles referred to herein is incorporated by reference for all purposes.70IPTS-200237393 1Attorney Docket No TGO-037WOEQUIVALENTS

[0294] The disclosure may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The foregoing embodiments are therefore to be considered in all respects illustrative rather than limiting the disclosure described herein. Various structural elements of the different embodiments and various disclosed method steps may be utilized in various combinations and permutations, and all such variants are to be considered forms of the disclosure.

[0295] The scope of this invention is to be detemiined by the appended claims in view of, but not limited to, the specific embodiments and illustrative examples described herein, and all changes that come within the meaning and range of equivalency of the claims are intended to be embraced therein.IPTS / 200237393.1

Claims

Attorney Docket No TGO-037WOCLAIMSWHAT IS CLAIMED IS:

1. A protein arginine methyltransferase 5 (PRMT5) inhibitor for use in treating a tumor in the central nervous system (CNS) in a subject in need thereof, wherein the PRMT5 inhibitor is a compound having the structural formula:■=NH2N. X,ZNHIf ' 9 I" ArWF 2 —? | — F.F Compound 1;wherein the compound is administered to the subject as a free base or a pharmaceutically acceptable salt or a composition thereof2. The compound for use of claim 1, wherein the tumor in the CNS is a glioma.3 The compound for use of claim 2, wherein the glioma is a glioblastoma.4 The compound for use of any one of claims 1 to 3, wherein the tumor in the CNS is MTAP-deficient.

5. The compound for use of any one of claims I to 4, wherein the tumor in the CNS is MTA-accumulating.

6. A protein arginine methyltransferase 5 (PRMT5) inhibitor for use in treating a cancer in a subject in need thereof, wherein the PRMT5 inhibitor is a compound having the structural formula:Attorney Docket No TGO-037WOF Compound 1;wherein the compound is administered to the subject as a free base or a pharmaceutically acceptable salt or a composition thereof.

7. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of claim 6, wherein the cancer comprises a solid tumor with MTAP-deficiency.

8. The protein arginine methyl transferase 5 (PRMT5) inhibitor for use of claim 6 or 7, wherein the cancer comprises glioma (e.g., glioblastoma), non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma.

9. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of claims 1 to 8, wherein the PRMT5 inhibitor is administered systemically.

10. The protein argmine methyltransferase 5 (PRMT5) inhibitor for use of any one of claims 1 to 9, wherein the PRMT5 inhibitor is administered orally.

11. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of claims 1 to 10, wherein the PRMT5 inhibitor is administered twice daily (BID).

12. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of claims 1 to 11, wherein the subject also receives a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor having the structural formula:1, NH h 1 y —- v-wCompound 2;wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.73IPTS'200237393.1Attorney Docket No TGO-037WO13. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of claim 12, wherein the CDK 4 / 6 inhibitor is administered systemically.

14. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of claim 12 or 13, wherein the CDK 4 / 6 inhibitor is administered orally.

15. A protein arginine methyltransferase 5 (PRMT5) inhibitor for use in combination with a cyclin-dependent kinase (CDK) 4 / 6 inhibitor in treating a cancer in a subject in need thereof, wherein:the PRMT5 inhibitor is a compound having the structural formula:F Compound 1;wherein Compound 1 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof; andthe CDK4 / 6 inhibitor is a compound having the structural formula:Compound 2;wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

1. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of claim 15, wherein the cancer comprises a solid tumor with MTAP-deficiency.

17. The protein arginine methyltransferase 5 (PRMT5) inhibitor of claim 15 or 16, wherein the cancer comprises glioma (c.g., glioblastoma), non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma.74IPTS'200237393.1Attorney Docket No TGO-037WO18. A protein arginine methyltransferase 5 (PRMT5) inhibitor for use in combination with a cyclin-dependent kinase (CDK) 4 / 6 inhibitor in treating a tumor in the central nervous system (CNS) in a subject in need thereof, wherein:the PRMT5 inhibitor is a compound having the structural formula:,=NlA"? IB° A*F Compound 1;wherein Compound 1 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof; andthe CDK4 / 6 inhibitor is a compound having the structural formula:Fwherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

19. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of claim 18, wherein the tumor in the CNS is a glioma.

20. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of claim 19, wherein the glioma is a glioblastoma.

21. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of claims 18 to 20, wherein the tumor m the CNS is MTAP-deficient22. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of claims 18 to 21, wherein the tumor in the CNS is MTA-accumulating.75IPTS'200237393.1Attorney Docket No TGO-037WO23. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of claims 15 to 22, wherein the PRMT5 inhibitor is administered systemically.

24. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of claims 15 to 23, wherein the PRMT5 inhibitor is administered orally.

25. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of claims 15 to 24, wherein the PRMT5 inhibitor is administered twice daily.

26. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of claims 15 to 25, wherein the CDK4 / 6 inhibitor is administered systemically.

27. The protein arginine methyltransferase 5 (PRMT5) inhibitor for use of any one of claims 15 to 26, wherein the CDK 4 / 6 inhibitor is administered orally,28. A method of treating a tumor in the central nervous system (CNS) in a subject in need thereof, the method comprising administering to the subj ect a therapeutically effective amount of a protein arginine methyltransferase 5 (PRMT5) inhibitor, wherein the PRMT5 inhibitor is a compound having the structural formula:F Compound 1:wherein the compound is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

29. The method of claim 28, wherein the tumor in the CNS is a glioma.

30. The method of claim 29. wherein the glioma is a glioblastoma.

31. The method of any one of claims 28 to 30, wherein the tumor in the CNS is MTAP-defi cient,Attorney Docket No TGO-037WO32. The method of any one of claims 28 to 31, wherein the tumor in the CNS is MTA-accumulating.

33. A method of treating a cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a protein arginine methyltransferase 5 (PRMT5) inhibitor, wherein the PRMT5 inhibitor is a compound having the structural formula:F Compound 1;wherein the compound is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

34. The method of claim 33, wherein the cancer comprises a solid tumor with MTAP- deficiency.

35. The method of claim 33 or 34, wherein the cancer comprises glioma (e.g, glioblastoma), non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma.

36. The method of any one of claims 28 to 35. wherein the PRMT5 inhibitor is administered systemically.

37. The method of any one of claims 28 to 36, wherein the PRMT5 inhibitor is administered orally.

38. The method of any one of claims 28-37, wherein the PRMT5 inhibitor is administered twice daily (BID).

39. The method of any one of claims 28 to 38, wherein the method further comprises administering to the subject a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor having the structural formula:Attorney Docket No TGO-037WOCompound 2;wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

40. The method of any one of claims 28 to 38, wherein the subject also receives a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor having the structural formula:f;» Y'V- Y""i 1hCompound 2;wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

41. The method of claim 39 or 40, wherein the CDK4 / 6 inhibitor is administered systemically.

42. The method of any one of claims 39 to 41, wherein the CDK 4 / 6 inhibitor is administered orally.

43. A method of treating a cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effecti ve amount of a protein arginine methyl transferase 5 (PRMT5) inhibitor and a therapeutically effective amount of a cyclin- dependent kinase (CDK) 4 / 6 inhibitor.wherein the PRMT5 inhibitor is a compound having the structural formula:IPTS'200237393.1Attorney Docket No TGO-037WOf-'NH,f< NH' ¥ " Y 9 IN-AAYNY"H0F Compound 1;wherein the CDK4 / 6 inhibitor is a compound having the structural formula:wherein Compound 1 and Compound 2 are each administered as a free base or a pharmaceutically acceptable salt or a composition thereof44. The method of claim 43, wherein the cancer comprises a solid tumor with MTAP-deficiency.

45. The method of claim 43 or 44, wherein the cancer comprises glioma (e.g., glioblastoma), non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma.

46. A method of treating a tumor in the CNS in a subject m need thereof, the method comprising administering to the subject a therapeutically effective amount of a protein arginine meth ltransferase 5 (PRMT5) inhibitor and a therapeutically effective amount of a cy elm-dependent kinase (CDK) 4 / 6 inhibitor,wherein the PRMT5 inhibitor is a compound having the structural formula:F Compound T.IPTS'200237393.1Attorney Docket No TGO-037WOwherein the CDK4 / 6 inhibitor is a compound having the structural formula:wherein Compound 1 and Compound 2 are each administered as a free base or a pharmaceutically acceptable salt or a composition thereof47. The method of claim 46, wherein the tumor in the CNS is a glioma,48. The method of claim 47, wherein the glioma is a glioblastoma.

49. The method of any one of claims 46 to 48, wherein the tumor in the CNS is MTAP- deficient.

50. The method of any one of claims 46 to 49, wherein the tumor in the CNS is MTA-accumulating.

51. The method of any one of claims 43 to 50. wherein the PRMT5 inhibitor is administered systemically.

52. The method of any one of claims 43 to 51, wherein the PRMT5 inhibitor is administered orally.

53. The method of any one of claims 43-52, wherein the PRMT5 inhibitor is administered twice daily (BID).

54. The method of any one of claims 43 to 53, wherein the CDK4 / 6 inhibitor is administered systemically.

55. The method of any one of claims 43 to 54. wherein the CDK 4 / 6 inhibitor is administered orally.

56. Use of a protein arginine methyltransferase 5 (PRMT5) inhibitor in the manufacture of a medicament for treating a tumor in the central nervous system (CNS) in a subject in need thereof, wherein the PRMT5 inhibitor is a compound having the structural formula:80IPTS'200237393.1Attorney Docket No TGO-037WOF Compound 1;wherein the compound is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof.

57. The use of claim 56, wherein the tumor in the CNS is a glioma.

58. The use of claim 57, wherein the glioma is a glioblastoma.

59. The use of any one of claims 56 to 58, wherein the tumor in the CNS is MTAP-deficient.

60. The use of any one of claims 56 to 59, wherein the tumor in the CNS is MTA-accumulating.

61. Use of a protein arginine methyltransferase 5 (PRMT5) inhibitor in the manufacture of a medicament for treating a cancer in a subject in need thereof, wherein the PRMT5 inhibitor is a compound having the structural formula:F Compound 1;wherein the compound is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof.

62. The use of claim 61, wherein the cancer comprises a solid tumor with MTAP-deficiency.

63. The use of claim 61 or 62, wherein the cancer comprises glioma (e.g., glioblastoma), nonsmall cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma.Attorney Docket No TGO-037WO64. The use of any one of claims 56 to 63. wherein the medicament is configured for systemic administration.

65. The use of any one of claims 56 to 64. wherein the medicament is configured for oral administration66. The use of any one of claims 56 to 65, wherein the medicament is configured for twice daily (BID) administration.

67. The use of any one of claims 56 to 66, wherein the subject also receives (or wherein the medicament is configured for administration with) a therapeutically effective amount of a cyclin-dependent kinase (CDK) 4 / 6 inhibitor having the structural formula:Compound 2;wherein Compound 2 is administered as a free base or a pharmaceutically acceptable salt or a composition thereof.

68. The use of claim 67, wherein the CDK 4 / 6 inhibitor is administered systemically.

69. The use of claim 67 or 68, wherein the CDK 4 / 6 inhibitor is administered orally, 70. Use of a protein arginine methyltransferase 5 (PRMT5) inhibitor in combination with a cyclin-dependent kinase (CDK) 4 / 6 inhibitor in the manufacture of a medicament for treating a cancer in a subject in need thereof, wherein:the PRMT5 inhibitor is a compound having the structural formula:F-- -~FF Compound 1:IPTS'200237393.1Attorney Docket No TGO-037WOwherein Compound 1 is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof; andthe CDK4 / 6 inhibitor is a compound having the structural formula:Compound 2;wherein Compound 2 is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof.

71. Use of a protein arginine methyltransferase 5 (PRMT5) inhibitor and a cyclin-dependent kinase (CDK) 4 / 6 inhibitor in the manufacture of medicaments for treating a cancer in a subject in need thereof in combination wherein:the PRMT5 inhibitor is a compound having the structural formula:F Compound 1;wherein Compound 1 is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof; andthe CDK4 / 6 inhibitor is a compound having the structural formula:Compound 2;83IPTS'200237393.1Attorney Docket No TGO-037WOwherein Compound 2 is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof and wherein the medicaments can be administered concomitantly or sequentially.

72. The use of claim 70 or 71, wherein the cancer comprises a solid tumor with MTAP-deficiency.

73. The use of any one of claims 70-72, wherein the cancer comprises glioma (e.g.. glioblastoma), non-small cell lung cancer, pancreatic cancer, bladder cancer or cholangiocarcinoma.

74. Use of a protein arginine methyltransferase 5 (PRMT5) inhibitor in combination with a cyclin-dependent kinase (CDK) 4 / 6 inhibitor in the manufacture of a medicament for treating a tumor in the central nervous system (CNS) in a subject in need thereof, wherein:the PRMT5 inhibitor is a compound having the structural formula:F Compound 1;wherein Compound 1 is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof; andthe CDK4 / 6 inhibitor is a compound having the structural formula:' Compound 2;wherein Compound 2 is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof.IPTS'200237393.1Attorney Docket No TGO-037WO75. Use of a protein arginine methyltransferase 5 (PRMT5) inhibitor and a cyclin-dependent kinase (CDK) 4 / 6 inhibitor in the manufacture of medicaments for treating a tumor in the central nervous system (CNS) in a subject in need thereof in combination wherein:the PRMT5 inhibitor is a compound having the structural formula:,=NH2N^k 'NHjA"? I" 11 JF Compound 1;wherein Compound 1 is present in the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof; andthe CDK4 / 6 inhibitor is a compound having the structural formula:wherein Compound 2 is present m the medicament as a free base or a pharmaceutically acceptable salt or a composition thereof and wherein the medicaments can be administered concomitantly or sequentially.

76. The use of claim 74 or 75, wherein the tumor in the CNS is a glioma.

77. The use of claim 76, wherein the glioma is a glioblastoma.

78. The use of any one of claims 74 to 77. wherein the tumor in the CNS is MTAP-deficient.

79. The use of any one of claims 74-78, wherein the tumor in the CNS is MTA-accumulating.

80. The use of any one of claims 70 to 79. wherein the PRMT5 inhibitor medicament is configured for systemic administration85IPTS'200237393.1Attorney Docket No TGO-037WO81. The use of any one of claims 70 to 80, wherein the PRMT5 inhibitor medicament is configured for oral administration.

82. The use of any one of claims 70 to 81. wherein the PRMT5 inhibitor medicament is configured for twice daily administration.

83. The use of any one of claims 70 to 82, wherein the CDK4 / 6 inhibitor medicament is configured for systemic administration.

84. The use of any one of claims 70 to 83, wherein the CDK4 / 6 inhibitor medicament is configured for oral administration.IPTS / 200237393.1

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