Treatment of cancer using MTA-cooperative PRMT5 inhibitors

Abstract

Described herein is a method for treating cancer in a patient, comprising administering a PRMT5 inhibitor. Also provided is a method for treating cancer in a patient, comprising administering a PRMT5 inhibitor and a standard treatment regimen.

Description

[Background technology]

[0001] Epigenetic regulation of gene expression is a key biological determinant of protein production and cell differentiation and plays a key pathogenetic role in many human diseases. Epigenetic regulation involves the genetic modification of genetic material without altering its nucleotide sequence. Typically, epigenetic regulation is mediated by selective and reversible modifications (e.g., methylation) of DNA and proteins (e.g., histones), thereby controlling the conformational transition of chromatin between transcriptionally active and inactive states. These covalent modifications can be controlled by enzymes such as methyltransferases (e.g., PRMT5), many of which are associated with specific genetic alterations that can cause human disease. PRMT5 influences diseases such as proliferative, metabolic, and hematological disorders.

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

[0003] In one embodiment, there is provided a method of treating cancer in a patient in need thereof, comprising administering to the patient an amount of a PRMT5 inhibitor ranging from 40 mg to 2000 mg, wherein the PRMT5 inhibitor is a compound of formula I or compound B: [ka] (In the formula, X 1 is NH, N(C1-C6 alkyl), O or S, X 2 is N(C1-C6 alkyl), O or S, Y 2 is H, C1-C6 alkyl or C1-C6 haloalkyl, Z 1 and Z 2 each is independently H, F, or C1-C6 alkyl, and Z 3 , Z 4 , Z 5 and Z 6 each is independently H, C1-C6 alkyl, or chloride. or a pharmaceutically acceptable salt thereof.

[0004] In another aspect, there is provided a method of treating cancer in a patient in need thereof, comprising: (a) a PRMT5 inhibitor in an amount ranging from 40 mg to 2000 mg, the compound being represented by Formula 1 or compound (B) [ka] (In the formula, X 1 is NH, N(C1-C6 alkyl), O or S, X 2 is N(C1-C6 alkyl), O or S, Y 2 is H, C1-C6 alkyl or C1-C6 haloalkyl, Z1 and Z 2 each is independently H, F, or C1-C6 alkyl, and Z 3 , Z 4 , Z 5 and Z 6 each is independently H, C1-C6 alkyl, or chloride. or a pharmaceutically acceptable salt thereof; and (b) Standard of care therapy for the treatment of cancer; to a patient. [Brief explanation of the drawings]

[0005] [Figure 1] Viability (A and B) and SDMA signaling (C and D) in MTAP-null or wild-type HAP1 (A / C) and HCT116 (B / D) cells after exposure to Compound B. WT and MTAP-null cells were treated with Compound B or DMSO-only control for 6 days. Viability was measured by CellTiter-Glo, and cooperativity was determined as follows: cooperativity = WT IC50 / MTAP-null IC50. (Mean ± standard deviation, n = 3). Global SDMA levels in HAP1 WT and MTAP-null cells were assessed by ELISA assay after 3 days of treatment with Compound B. Global SDMA levels in HCT116 WT and MTAP-null cells were assessed by in-cell imaging assay after 4 days of treatment with Compound B. [Figure 2] ELISA analysis of HCT116 MTAP WT and HCT116 MTAP null contralateral tumor cells. Female athymic nude mice were dosed with vehicle or Compound B. Mice were dosed a total of four times, and tumors were harvested 4 hours after the last dose. Percent inhibition relative to the corresponding vehicle was recorded. Data represent mean ± standard deviation, n=3 per group. Statistics: Dunnett's one-way ANOVA with comparison to control p=0.0213*, p<0.0001****. [Figure 3]Compound B induces significant anti-tumor growth inhibition in patient-derived xenografts of endogenous MTAP-null pancreas (A) and esophagus (B). Female NOD / SCID mice were implanted with PA5415 (pancreatic tumor) or ES11082 (esophageal tumor). Vehicle and Compound B were administered orally (po) once daily for the study period. Data represent mean ± standard error, n=10 per group. B.) Statistics: P-values ​​were determined by a linear mixed-effects model with Dunnett's comparison to control, ****p<0.0001. [Figure 4] Tumor growth inhibition in the presence of Compound B in an endogenous MTAP-null patient-derived xenograft model. (From left to right): pancreas, esophagus, esophagus, pancreas, melanoma, lung, mixed Müllerian duct, lung, melanoma, lung, pancreas, lung, ovary, lung, gallbladder, lung, melanoma, melanoma, lung, melanoma, pancreas, melanoma, pancreas, brain, and pancreas. For each model, six female NOD / SCID mice were implanted with PDX. The mean tumor volume for each group was 100-200 mm3. Mice were assigned to two different study groups based on tumor volume and initiated with 100 mg / kg vehicle or Compound B. Data plotted represent TGI (tumor growth inhibition), n=3 per group. [Figure 5] Antitumor activity of Compound B in HCT116 MTAP-null xenografts (A) versus HCT116 wild-type xenografts (B). Female athymic nude mice were implanted with HCT116 MTAP-null or MTAP-WT tumors. Vehicle and Compound B were administered orally (po) once daily during the study period. Data represent group mean ± standard error, n=10. No effects on body weight were observed in either study. Statistics: P values ​​were determined by a linear mixed-effects model with comparison to Dunnett's control. A) **p<0.01, ****p<0.0001. B) p=NS. [Figure 6]Compound B exhibits preferential activity in endogenous MTAP-null tumor cell lines. Waterfall plot of mean IC50 values ​​across a panel of endogenous wild-type (n = 11) or MTAP-null (n = 15) cell lines treated with Compound B, an MTA-cooperative PRMT5 inhibitor. Cells were seeded in 96-well plates for optimal log-phase growth, incubated overnight, and then treated with compound across a 3-fold, 9-point serial dilution series for a 6-day viability assay (Cell-Titer-Glo; Promega). Signal intensities across the dilution series were normalized to the DMSO control (0.1%) and averaged across two biological replicates. IC50 values ​​were calculated using a four-parameter interpolated IC50 model (Prism). [Figure 7] 1 is a graph showing that the combination of Compound G with paclitaxel resulted in significant anti-tumor activity compared to single agents alone in H292 NSCLC xenografts. [Figure 8] 1 is a graph showing that the combination of Compound B with paclitaxel results in significant anti-tumor activity compared to single agents alone in H292 NSCLC xenografts. [Figure 9-1] Compound B preferentially inhibits cell viability of endogenous MTAP-null cancer cell lines compared to wild-type (A) DLBCL, (B) pancreatic, and (C) lung cancer cell lines. Representative dose-response curves of WT and MTAP-null DLBCL pancreatic and NSCLC cancer tumor cell lines treated with serial dilutions of Compound B for 6 days. The highest concentration tested was 10 μM (1 μM for DLBCL lines) with a total of nine 1:3 serial dilutions and a DMSO-only control. Cell viability was measured by CellTiter-Glo luminescence assay. (Mean ± standard deviation, n = 3). [Figure 9-2]Compound B preferentially inhibits cell viability of endogenous MTAP-null cancer cell lines compared to wild-type (A) DLBCL, (B) pancreatic, and (C) lung cancer cell lines. Representative dose-response curves of WT and MTAP-null DLBCL pancreatic and NSCLC cancer tumor cell lines treated with serial dilutions of Compound B for 6 days. The highest concentration tested was 10 μM (1 μM for DLBCL lines) with a total of nine 1:3 serial dilutions and a DMSO-only control. Cell viability was measured by CellTiter-Glo luminescence assay. (Mean ± standard deviation, n = 3). [Figure 10-1] Compound B exhibits significant antitumor activity in endogenous MTAP-null xenografts of DOHH-2 (A and B) and BxPC-3 (C and D). Female SCID mice were implanted with DOHH-2 or BxPC-3 tumors. Vehicle and Compound B were orally administered (po) once daily during the study period. Data represent group mean ± standard error, n = 10. Late-stage DOHH-2 or BxPC-3 tumors were harvested for ELISA analysis of SDMA. Data represent mean ± standard deviation, n = 5 per group. Statistics: P values ​​were determined by a linear mixed-effects model with Dunnett's comparison to control; **p < 0.01, ****p < 0.0001. [Figure 10-2] Compound B exhibits significant antitumor activity in endogenous MTAP-null xenografts of DOHH-2 (A and B) and BxPC-3 (C and D). Female SCID mice were implanted with DOHH-2 or BxPC-3 tumors. Vehicle and Compound B were orally administered (po) once daily during the study period. Data represent group mean ± standard error, n = 10. Late-stage DOHH-2 or BxPC-3 tumors were harvested for ELISA analysis of SDMA. Data represent mean ± standard deviation, n = 5 per group. Statistics: P values ​​were determined by a linear mixed-effects model with Dunnett's comparison to control; **p < 0.01, ****p < 0.0001. [Figure 11-1]Compound B induces cell cycle arrest and augments the DNA damage response in DOHH-2 tumors without affecting circulating blood cells (A-D). Female SCID mice were implanted with DOHH-2 tumors. When tumors reached 200 mm3, mice were treated with 100 mg / kg of either vehicle or Compound B. Mice were orally administered (po) once daily for 10 days. Data represent group mean ± standard error, n=10. A.) End-stage DOHH-2 tumor volume. B.) Tumors isolated from vehicle-treated or Compound B-treated mice pulsed with Brdu (4 h) were co-stained with DAPI and analyzed by flow cytometry. C.) Dissociated tumors were stained for γH2AX, MFI (mean fluorescence intensity). D.) Cardiac blood samples were collected and analyzed by ADVIA. [Figure 11-2] Compound B induces cell cycle arrest and augments the DNA damage response in DOHH-2 tumors without affecting circulating blood cells (A-D). Female SCID mice were implanted with DOHH-2 tumors. When tumors reached 200 mm3, mice were treated with 100 mg / kg of either vehicle or Compound B. Mice were orally administered (po) once daily for 10 days. Data represent group mean ± standard error, n=10. A.) End-stage DOHH-2 tumor volume. B.) Tumors isolated from vehicle-treated or Compound B-treated mice pulsed with Brdu (4 h) were co-stained with DAPI and analyzed by flow cytometry. C.) Dissociated tumors were stained for γH2AX, MFI (mean fluorescence intensity). D.) Cardiac blood samples were collected and analyzed by ADVIA. [Figure 11-3]Compound B induces cell cycle arrest and augments the DNA damage response in DOHH-2 tumors without affecting circulating blood cells (A-D). Female SCID mice were implanted with DOHH-2 tumors. When tumors reached 200 mm3, mice were treated with 100 mg / kg of either vehicle or Compound B. Mice were orally administered (po) once daily for 10 days. Data represent group mean ± standard error, n=10. A.) End-stage DOHH-2 tumor volume. B.) Tumors isolated from vehicle-treated or Compound B-treated mice pulsed with Brdu (4 h) were co-stained with DAPI and analyzed by flow cytometry. C.) Dissociated tumors were stained for γH2AX, MFI (mean fluorescence intensity). D.) Cardiac blood samples were collected and analyzed by ADVIA. [Figure 12] 1 is a dose matrix showing the combination index (CI) scores for Compound B in combination with paclitaxel. [Figure 13] 1 is a dose matrix showing the combination index (CI) scores for Compound B in combination with carboplatin. [Figure 14A] 14A and 14B are graphs showing that the combination of Compound B with paclitaxel (FIG. 14A) and the combination of Compound B with carboplatin (FIG. 14B) result in significant anti-tumor cell proliferation activity compared to paclitaxel or carboplatin alone in NSCLC (H292) cell lines. [Figure 14B] 14A and 14B are graphs showing that the combination of Compound B with paclitaxel (FIG. 14A) and the combination of Compound B with carboplatin (FIG. 14B) result in significant anti-tumor cell proliferation activity compared to paclitaxel or carboplatin alone in NSCLC (H292) cell lines. [Figure 15A] 15A and 15B are graphs showing that the combination of Compound B with paclitaxel (FIG. 15A) and the combination of Compound B with carboplatin (FIG. 15B) result in significant antitumor activity compared to paclitaxel or carboplatin alone in H292 NSCLC xenografts. [Figure 15B]15A and 15B are graphs showing that the combination of Compound B with paclitaxel (FIG. 15A) and the combination of Compound B with carboplatin (FIG. 15B) result in significant antitumor activity compared to paclitaxel or carboplatin alone in H292 NSCLC xenografts. DETAILED DESCRIPTION OF THE INVENTION

[0006] The present disclosure provides a method of treating cancer in a patient, comprising administering a PRMT5 inhibitor, wherein the PRMT5 inhibitor is a compound of Formula I or Compound B: [ka] (In the formula, X 1 is NH, N(C1-C6 alkyl), O or S, X 2 is N(C1-C6 alkyl), O or S, Y 2 is H, C1-C6 alkyl or C1-C6 haloalkyl, Z 1 and Z 2 each is independently H, F, or C1-C6 alkyl, and Z 3 , Z 4 , Z 5 and Z 6 each is independently H, C1-C6 alkyl, or chloride. or a pharmaceutically acceptable salt thereof.

[0007] In some embodiments, the PRMT5 inhibitor has formula (S)-I: [ka] or a pharmaceutically acceptable salt thereof.

[0008] In some embodiments, X 1 is O. In some embodiments, Z 1 and Z 2are each H. In some embodiments, X 2 is O. In some embodiments, Z 3 , Z 4 , Z 5 and Z 6 Each of Y is H. In some embodiments, Y 2 is C1-C6 haloalkyl. In some embodiments, Y 2 is CF3.

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

[0010] In some embodiments, the PRMT5 inhibitor has Formula A: [ka] or a salt thereof.

[0011] In some embodiments, the PRMT5 inhibitor has formula G: [ka] or a salt thereof.

[0012] Pharmaceutically acceptable salts of the compounds described herein include those derived from suitable inorganic and organic acids and bases.

[0013] Combination therapy In some embodiments, the method further comprises administering a standard of care therapy to the patient as a combination therapy. As used herein, the term "combination therapy" refers to the administration of two or more therapeutic agents (e.g., a PRMT5 inhibitor as described in the present disclosure and a standard of care therapy (e.g., chemotherapy)) to treat cancer. Such administration encompasses co-administration of these therapeutic agents in a substantially simultaneous manner, such as in a single capsule with a fixed ratio of active ingredients. Alternatively, such administration encompasses simultaneous administration in multiple or separate containers (e.g., tablets, capsules, powders, and liquids) for each active ingredient. The powders and / or liquids can be reconstituted or diluted to the desired dose before administration. In addition, such administration also encompasses the use of each type of therapeutic agent sequentially, either at approximately the same time or at different times.

[0014] In some embodiments, the chemotherapy is platinum-based chemotherapy, i.e., includes a platinum agent. Platinum agents (such as carboplatin, oxaliplatin, cisplatin, nedaplatin, satraplatin, lobaplatin, triplatin tetranitrate, picoplatin, ProLindac™ (AP5346), aroplatin, and phenanthriplatin) are widely used antitumor drugs that cause crosslinking of DNA as monoadducts, interstrand crosslinks, intrastrand crosslinks, or DNA-protein crosslinks.

[0015] Carboplatin is a platinum alkylating agent that covalently binds to DNA and interferes with DNA function by creating intrastrand DNA crosslinks. Platinum-based chemotherapy is the first-line regimen for patients with advanced or metastatic non-squamous NSCLC without genomic EGFR or ALK tumor abnormalities, with or without immunotherapy. Exemplary chemotherapeutic agents that can be combined with platinum typically include, but are not limited to, pemetrexed, taxanes (paclitaxel, nab-paclitaxel, or docetaxel), and etoposide, or any combination of the foregoing.

[0016] Carboplatin has the molecular formula CH 12 N2O4Pt is a water-soluble platinum complex with a molecular weight of 373.26. Carboplatin is registered under CAS Registry Number 41575-94-4 and is commercially available as PARAPLATIN®, BLASTOCARB®, BLASTOPLATIN®, CARBOKEM®, CARBOMAX®, PARAPLATIN®, CARBOPA®, KARPLAT®, etc. Complete information regarding the preparation, formulation, dosage, and administration schedule of carboplatin can be found in the local package insert (in the United States, see, for example, CARBOplatin Injection, US Prescribing Information, Fresenius KABI, Lake Zurich, Illinois, 60047 (revision 5 / 2021), which is incorporated herein by reference in its entirety).

[0017] In some embodiments, the chemotherapeutic agent is an antifolate chemotherapeutic agent. In some embodiments, the chemotherapy is pemetrexed or a pharmaceutically acceptable salt thereof. In some embodiments, the chemotherapy is pemetrexed. In some embodiments, the chemotherapy is pemetrexed. In some embodiments, the chemotherapy is pemetrexed disodium. Pemetrexed (N-[4-[2-(2-amino-4,7-dihydro-4-oxo-1H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]-L-glutamic acid)) is a folate analog metabolic inhibitor that inhibits folate-dependent metabolic processes essential for cell replication. Pemetrexed, in combination with pembrolizumab and platinum chemotherapy, has been approved by the FDA for the initial treatment of patients with metastatic non-squamous NSCLC without EGFR or ALK tumor genomic aberrations. It has also been approved in combination with cisplatin for the initial treatment of patients with locally advanced or metastatic non-squamous NSCLC. In the maintenance setting, pemetrexed, as a single agent, is approved for the treatment of patients with locally advanced or metastatic non-squamous NSCLC whose disease has not progressed after four 3-week cycles of platinum-based first-line chemotherapy. It is also approved, as a single agent, for the treatment of patients with recurrent, metastatic non-squamous NSCLC after prior chemotherapy. Pemetrexed is commercially available as ALIMTA®. Complete information regarding the formulation, dosage, and administration schedule of pemetrexed can be found in the local package insert (in the United States, see, e.g., ALIMTA® US Prescribing Information, Lilly USA, LLC, Indianapolis, Indiana 46285 (revision 1 / 2019), which is incorporated herein by reference in its entirety).

[0018] In some embodiments, the chemotherapy is a taxane. Exemplary taxanes include, but are not limited to, paclitaxel (TAXOL®), a cremophor-free albumin-modified nanoparticle formulation of paclitaxel or nab-paclitaxel (ABRAXANE®), and docetaxel (TAXOTERE®).

[0019] Paclitaxel is a semisynthetic taxane, a class of anticancer drugs that binds to beta-tubulin, thereby stabilizing microtubules, arresting the cell cycle, and inducing apoptosis. It is administered intravenously at 200 mg / m over 3 hours every 3 weeks in combination with carboplatin. 2 Paclitaxel has become the standard of care option for the treatment of patients with previously untreated, advanced or metastatic NSCLC who perform well (Schiller JH et al, 2002). Docetaxel is a semisynthetic taxane, a class of anticancer drugs that binds to beta-tubulin, thereby stabilizing microtubules, arresting the cell cycle and inducing apoptosis. Docetaxel is administered intravenously over 1 hour every 3 weeks as monotherapy at a dose of 75 mg / m 2 Docetaxel is approved by the FDA for the treatment of patients with locally advanced or metastatic NSCLC after failure of prior platinum-based chemotherapy.

[0020] Complete information regarding the preparation, dispensing, dosage, and administration schedule of paclitaxel (TAXOL®) can be found in the local package insert (in the United States, see, e.g., TAXOL® (paclitaxel) INJECTION US Prescribing Information, Bristol-Myers Squibb Company, Princeton, New Jersey, 08543 (revision 4 / 2011), which is incorporated herein by reference in its entirety). Complete information regarding the preparation, dispensing, dosage, and administration schedule of nab-paclitaxel (ABRAXANE®) can be found in the local package insert (in the United States, see, e.g., ABRAXANE® US Prescribing Information, Bristol-Myers Squibb Company, New Jersey, 08543 (revision 8 / 2020), which is incorporated herein by reference in its entirety). Complete information regarding the preparation, dispensing, dosage and administration schedule of docetaxel can be found in the local package insert (in the United States, see, e.g., Docetaxel Injection US Prescribing Information, Sandoz, Princeton, New Jersey, 08540 (revision 3 / 2012)), which is incorporated herein by reference in its entirety).

[0021] In some embodiments, the chemotherapeutic agent is docetaxel, paclitaxel, carboplatin, gemcitabine, irinotecan, 5-fluororacil, or pemetrexed. In some embodiments, the method comprises administering carboplatin to the patient. In some embodiments, the method comprises administering docetaxel to the patient. In some embodiments, the method comprises administering paclitaxel to the patient. In some embodiments, the method comprises administering pemetrexed to the patient. In some embodiments, the method comprises administering gemcitabine to the patient. In some embodiments, the method comprises administering irinotecan to the patient. In some embodiments, the method comprises administering 5-fluororacil to the patient.

[0022] Dosing regimen A "therapeutically effective amount" of a PRMT5 inhibitor refers to an amount effective to treat, prevent progression of, or alleviate existing symptoms in a patient receiving treatment. Determining an effective amount is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein. Generally, a "therapeutically effective amount" refers to the amount of a PRMT5 inhibitor described herein that results in achieving a desired effect. For example, a therapeutically effective amount of a PRMT5 inhibitor described herein reduces MTAP activity by at least 5% compared to a control, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90% compared to a control.

[0023] Determining optimal ranges for effective amounts of compounds is within the skill of the art, although this will vary depending on individual needs. For example, for administration to humans in treating the conditions and disorders identified herein, a typical dosage of a compound as disclosed herein can be about 0.05 mg / kg / day to about 50 mg / kg / day, e.g., at least 0.05 mg / kg, at least 0.08 mg / kg, at least 0.1 mg / kg, at least 0.2 mg / kg, at least 0.3 mg / kg, at least 0.4 mg / kg, or at least 0.5 mg / kg, e.g., 50 mg / kg or less, 40 mg / kg or less, 30 mg / kg or less, 20 mg / kg or less, or 10 mg / kg or less, which can be about 2.5 mg / day (0.5 mg / kg x 5 kg) to about 5000 mg / day (50 mg / kg x 100 kg). For example, the dosage of the compound may be from about 0.1 mg / kg / day to about 50 mg / kg / day, from about 0.05 mg / kg / day to about 10 mg / kg / day, from about 0.05 mg / kg / day to about 5 mg / kg / day, from about 0.05 mg / kg / day to about 3 mg / kg / day, from about 0.07 mg / kg / day to about 3 mg / kg / day, from about 0.09 mg / kg / day to about 3 mg / kg / day, from about 0.05 mg / kg / day to about 0.1 mg / kg / day, from about 0.1 mg / kg / day to about 1 mg / kg / day, or from about 1 mg / kg / day to about 10 mg / kg / day. daily, about 1 mg / kg / day to about 5 mg / kg / day, about 1 mg / kg / day to about 3 mg / kg / day, about 1 mg / day to about 2000 mg / day, about 20 mg / day to about 1800 mg / day, about 40 mg / day to about 800 mg / day, about 20 mg / day to about 700 mg / day, about 30 mg / day to about 600 mg / day, about 40 mg / day to about 500 mg / day, about 50 mg / day to about 400 mg / day, about 60 mg / day to about 300 mg / day, about 70 mg / day to about 200 mg / day, or about 80 mg / day to about 100 mg / day.

[0024] In certain embodiments, the PRMT5 inhibitors described herein are orally administered once daily to a patient in need thereof. Contemplated "patients" or "subjects" include, but are not limited to, humans (i.e., males or females of any age group, such as pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or elderly adults)) and / or non-human animals, such as mammals, such as primates (e.g., cynomolgus monkeys, rhesus monkeys), cows, 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.

[0025] In some embodiments, the patient has been treated with a previous line of therapy, i.e., the therapy of the methods disclosed herein is second (or higher) line therapy. In some embodiments, the patient has been previously treated with chemotherapy before being treated with a PRMT5 inhibitor (as disclosed in the methods herein). In some embodiments, the patient has been previously treated with a PD1 inhibitor before being treated with a PRMT5 inhibitor (as disclosed in the methods herein). In some embodiments, the patient has been previously treated with a PDL1 inhibitor before being treated with a PRMT5 inhibitor (as disclosed in the methods herein).

[0026] In some cases, the patient is administered a total daily dose of 40 mg, 120 mg, 240 mg, 480 mg, 800 mg, 960 mg, 1600 mg, or 2000 mg of a PRMT5 inhibitor.

[0027] In some embodiments, the methods comprise administering a PRMT5 inhibitor described herein in an amount ranging from 40 mg to 2000 mg, hi some embodiments, the methods comprise administering 40 mg, 120 mg, 240 mg, 480 mg, 800 mg, 960 mg, 1600 mg, or 2000 mg of a PRMT5 inhibitor to a patient once daily.

[0028] In some embodiments, the method comprises administering docetaxel to the patient. 2 of docetaxel administered IV every 3 weeks. In some embodiments, the methods described herein comprise administering (a) 40 mg of a PRMT5 inhibitor daily, and (b) 75 mg / m 2 of docetaxel administered IV every 3 weeks. In some embodiments, the methods described herein comprise administering (a) 120 mg of a PRMT5 inhibitor daily, and (b) 75 mg / m 2 of docetaxel administered IV every 3 weeks. In some embodiments, the methods described herein comprise administering (a) 240 mg of a PRMT5 inhibitor daily, and (b) 75 mg / m 2 of docetaxel administered IV every 3 weeks. In some embodiments, the methods described herein comprise administering (a) 480 mg of a PRMT5 inhibitor daily, and (b) 75 mg / m 2 of docetaxel administered IV every 3 weeks. In some embodiments, the methods described herein comprise administering (a) 800 mg of a PRMT5 inhibitor daily, and (b) 75 mg / m 2 of docetaxel administered IV every 3 weeks. In some embodiments, the methods described herein comprise administering (a) 2000 mg of a PRMT5 inhibitor daily, and (b) 75 mg / m 2 of docetaxel administered IV every three weeks.

[0029] In some embodiments, the method comprises administering carboplatin to the patient. In some embodiments, the method comprises administering carboplatin at an AUC5 (or AUC6) by IV administration every three weeks. In some embodiments, the method described herein comprises administering to the patient (a) 40 mg of a PRMT5 inhibitor daily, and (b) carboplatin at an AUC5 (or AUC6) by IV administration every three weeks. In some embodiments, the method described herein comprises administering to the patient (a) 120 mg of a PRMT5 inhibitor daily, and (b) carboplatin at an AUC5 (or AUC6) by IV administration every three weeks. In some embodiments, the method described herein comprises administering to the patient (a) 240 mg of a PRMT5 inhibitor daily, and (b) carboplatin at an AUC5 (or AUC6) by IV administration every three weeks. In some embodiments, the methods described herein comprise administering to a patient (a) 480 mg of a PRMT5 inhibitor daily and (b) carboplatin at an AUC5 (or AUC6) via IV administration every three weeks. In some embodiments, the methods described herein comprise administering to a patient (a) 800 mg of a PRMT5 inhibitor daily and (b) carboplatin at an AUC5 (or AUC6) via IV administration every three weeks. In some embodiments, the methods described herein comprise administering to a patient (a) 2000 mg of a PRMT5 inhibitor daily and (b) carboplatin at an AUC5 (or AUC6) via IV administration every three weeks.

[0030] In some embodiments, the method comprises administering paclitaxel to the patient. In some embodiments, the method comprises administering paclitaxel at a dose of 100 mg / m 2 Paclitaxel was administered intravenously weekly (or 135 mg / m 2 of paclitaxel administered IV every 3 weeks. In some embodiments, the method comprises administering (a) 40 mg of a PRMT5 inhibitor daily and (b) 100 mg / m 2 Paclitaxel was administered intravenously weekly (or 135 mg / m 2of paclitaxel administered IV every 3 weeks to the patient. In some embodiments, the method comprises administering to the patient (a) 80 mg of a PRMT5 inhibitor daily, and (b) 100 mg / m 2 Paclitaxel was administered intravenously weekly (or 135 mg / m 2 of paclitaxel administered IV every 3 weeks to the patient. In some embodiments, the method comprises administering to the patient: (a) 120 mg of a PRMT5 inhibitor daily; and (b) 100 mg / m 2 of paclitaxel administered IV weekly (or 135 mg / m2 of paclitaxel administered IV every 3 weeks) to the patient. In some embodiments, the method comprises administering (a) 240 mg of a PRMT5 inhibitor daily and (b) 100 mg / m 2 Paclitaxel was administered intravenously weekly (or 135 mg / m 2 of paclitaxel administered IV every 3 weeks to the patient. In some embodiments, the method comprises administering to the patient (a) 800 mg of a PRMT5 inhibitor daily, and (b) 100 mg / m 2 Paclitaxel was administered intravenously weekly (or 135 mg / m 2 of paclitaxel administered IV every 3 weeks to the patient. In some embodiments, the method comprises administering to the patient: (a) 2000 mg of a PRMT5 inhibitor daily; and (b) 100 mg / m 2 Paclitaxel was administered intravenously weekly (or 135 mg / m 2 of paclitaxel (administered IV every 3 weeks) to the patient.

[0031] In some embodiments, the methods include administering pemetrexed to the patient. In some embodiments, the methods include administering 500 mg / m 2 of pemetrexed administered IV every three weeks. In some embodiments, the methods described herein comprise administering (a) 40 mg of a PRMT5 inhibitor daily, and (b) 500 mg / m 2of pemetrexed administered IV to a patient every three weeks. In some embodiments, the methods described herein include administering (a) 80 mg of a PRMT5 inhibitor daily, and (b) 500 mg / m 2 of pemetrexed administered IV every three weeks to a patient. In some embodiments, the methods described herein include administering (a) 120 mg of a PRMT5 inhibitor daily, and (b) 500 mg / m 2 of pemetrexed administered IV to a patient every three weeks. In some embodiments, the methods described herein include administering (a) 240 mg of a PRMT5 inhibitor daily, and (b) 500 mg / m 2 of pemetrexed administered IV to a patient every three weeks. In some embodiments, the methods described herein include administering (a) 480 mg of a PRMT5 inhibitor daily, and (b) 500 mg / m 2 of pemetrexed administered IV to a patient every three weeks. In some embodiments, the methods described herein comprise administering (a) 800 mg of a PRMT5 inhibitor daily, and (b) 500 mg / m 2 of pemetrexed administered IV to a patient every three weeks. In some embodiments, the methods described herein include administering (a) 2000 mg of a PRMT5 inhibitor daily, and (b) 500 mg / m 2 of pemetrexed administered intravenously to patients every three weeks.

[0032] In some embodiments, the methods include administering gemcitabine to the patient. In some embodiments, the methods include administering 1000 mg / m 2 of gemcitabine administered as an intravenous infusion over 30 minutes on days 1, 8, and 15 of each 28-day cycle. In some embodiments, the methods described herein comprise administering (a) 40 mg of a PRMT5 inhibitor daily, and (b) 1000 mg / m 2of gemcitabine administered to a patient as an intravenous infusion over 30 minutes on days 1, 8, and 15 of each 28-day cycle. In some embodiments, the methods described herein comprise administering (a) 80 mg of a PRMT5 inhibitor daily, and (b) 1000 mg / m 2 of gemcitabine administered to a patient as an intravenous infusion over 30 minutes on days 1, 8, and 15 of each 28-day cycle. In some embodiments, the methods described herein comprise administering (a) 120 mg of a PRMT5 inhibitor daily, and (b) 1000 mg / m 2 of gemcitabine administered to a patient as an intravenous infusion over 30 minutes on days 1, 8, and 15 of each 28-day cycle. In some embodiments, the methods described herein comprise administering (a) 480 mg of a PRMT5 inhibitor daily, and (b) 1000 mg / m 2 of gemcitabine administered to a patient as an intravenous infusion over 30 minutes on days 1, 8, and 15 of each 28-day cycle. In some embodiments, the methods described herein comprise administering (a) 800 mg of a PRMT5 inhibitor daily, and (b) 1000 mg / m 2 of gemcitabine administered to a patient as an intravenous infusion over 30 minutes on days 1, 8, and 15 of each 28-day cycle. In some embodiments, the methods described herein comprise administering (a) 2000 mg of a PRMT5 inhibitor daily, and (b) 1000 mg / m 2 of gemcitabine administered to the patient as an intravenous infusion over 30 minutes on days 1, 8, and 15 of each 28-day cycle.

[0033] In some embodiments, the method provides a 1250 mg / m 2 of gemcitabine administered as an intravenous infusion over 30 minutes on days 1 and 8 of each 21-day cycle. In some embodiments, the methods described herein comprise administering (a) 40 mg of a PRMT5 inhibitor daily, and (b) 1250 mg / m 2of gemcitabine administered to a patient by IV infusion over 30 minutes on days 1 and 8 of each 21-day cycle. In some embodiments, the methods described herein comprise administering (a) 80 mg of a PRMT5 inhibitor daily, and (b) 1250 mg / m 2 of gemcitabine administered to a patient by IV infusion over 30 minutes on days 1 and 8 of each 21-day cycle. In some embodiments, the methods described herein comprise administering (a) 120 mg of a PRMT5 inhibitor daily, and (b) 1250 mg / m 2 of gemcitabine administered to a patient by IV infusion over 30 minutes on days 1 and 8 of each 21-day cycle. In some embodiments, the methods described herein comprise administering (a) 480 mg of a PRMT5 inhibitor daily, and (b) 1250 mg / m 2 of gemcitabine administered to a patient by IV infusion over 30 minutes on days 1 and 8 of each 21-day cycle. In some embodiments, the methods described herein comprise administering (a) 800 mg of a PRMT5 inhibitor daily, and (b) 1250 mg / m 2 of gemcitabine administered to a patient by IV infusion over 30 minutes on days 1 and 8 of each 21-day cycle. In some embodiments, the methods described herein comprise administering (a) 2000 mg of a PRMT5 inhibitor daily, and (b) 1250 mg / m 2 of gemcitabine administered to patients by IV infusion over 30 minutes on days 1 and 8 of each 21-day cycle.

[0034] In some embodiments, the methods include administering irinotecan to the patient. In some embodiments, the methods include administering irinotecan at a dose of 180 mg / m 2 of irinotecan administered IV every two weeks. In some embodiments, the methods described herein comprise administering (a) 40 mg of a PRMT5 inhibitor daily, and (b) 180 mg / m 2of irinotecan administered IV to a patient every two weeks. In some embodiments, the methods described herein include administering (a) 80 mg of a PRMT5 inhibitor daily, and (b) 180 mg / m 2 of irinotecan administered IV every two weeks to a patient. In some embodiments, the methods described herein include administering (a) 120 mg of a PRMT5 inhibitor daily, and (b) 180 mg / m 2 of irinotecan administered IV to a patient every two weeks. In some embodiments, the methods described herein include administering (a) 480 mg of a PRMT5 inhibitor daily, and (b) 180 mg / m 2 of irinotecan administered IV every two weeks to a patient. In some embodiments, the methods described herein include administering (a) 800 mg of a PRMT5 inhibitor daily, and (b) 180 mg / m 2 of irinotecan administered IV to a patient every two weeks. In some embodiments, the methods described herein include administering (a) 2000 mg of a PRMT5 inhibitor daily, and (b) 180 mg / m 2 of irinotecan administered intravenously to patients every two weeks.

[0035] In some embodiments, the method provides a dose of 150 mg / m 2 of irinotecan administered IV every two weeks. In some embodiments, the methods described herein comprise administering (a) 40 mg of a PRMT5 inhibitor daily, and (b) 150 mg / m 2 of irinotecan administered IV to a patient every two weeks. In some embodiments, the methods described herein include administering (a) 80 mg of a PRMT5 inhibitor daily, and (b) 150 mg / m 2 of irinotecan administered IV every two weeks to a patient. In some embodiments, the methods described herein include administering (a) 120 mg of a PRMT5 inhibitor daily, and (b) 150 mg / m 2of irinotecan administered IV to a patient every two weeks. In some embodiments, the methods described herein include administering (a) 480 mg of a PRMT5 inhibitor daily, and (b) 150 mg / m 2 of irinotecan administered IV to a patient every two weeks. In some embodiments, the methods described herein include administering (a) 800 mg of a PRMT5 inhibitor daily, and (b) 150 mg / m 2 of irinotecan administered IV to a patient every two weeks. In some embodiments, the methods described herein include administering (a) 2000 mg of a PRMT5 inhibitor daily, and (b) 150 mg / m 2 of irinotecan administered intravenously to patients every two weeks.

[0036] In some embodiments, the method comprises administering 5-fluoracil to the patient. In some embodiments, the method comprises administering 400 mg / m 2 of 5-fluoracil as an IV bolus, followed by 2400–3000 mg / m 2 by IV continuous infusion for 46 hours. In some embodiments, the methods described herein comprise administering (a) 40 mg of a PRMT5 inhibitor daily, and (b) 400 mg / m 2 of 5-fluoracil as an IV bolus, followed by 2400–3000 mg / m 2 to a patient via IV continuous infusion for 46 hours. In some embodiments, the methods described herein comprise administering (a) 80 mg of a PRMT5 inhibitor daily, and (b) 400 mg / m 2 of 5-fluoracil as an IV bolus, followed by 2400–3000 mg / m 2 to a patient via IV continuous infusion for 46 hours. In some embodiments, the methods described herein comprise administering (a) 120 mg of a PRMT5 inhibitor daily, and (b) 400 mg / m 2 of 5-fluoracil as an IV bolus, followed by 2400–3000 mg / m 2to a patient via IV continuous infusion for 46 hours. In some embodiments, the methods described herein comprise administering (a) 240 mg of a PRMT5 inhibitor daily, and (b) 400 mg / m 2 of 5-fluoracil as an IV bolus, followed by 2400–3000 mg / m 2 to a patient via IV continuous infusion for 46 hours. In some embodiments, the methods described herein comprise administering (a) 800 mg of a PRMT5 inhibitor daily, and (b) 400 mg / m 2 of 5-fluoracil as an IV bolus, followed by 2400–3000 mg / m 2 by IV continuous infusion x 46 hours to a patient. In some embodiments, the methods described herein comprise administering (a) 2000 mg of a PRMT5 inhibitor daily, and (b) 400 mg / m 2 of 5-fluoracil as an IV bolus, followed by 2400–3000 mg / m 2 This involves administering the drug to the patient via IV continuous infusion for 46 hours.

[0037] In some embodiments, the method includes administering 5-fluoracil to the patient. In some embodiments, the method includes administering 1200 mg / m 2 of 5-fluoracil administered by IV continuous infusion for 44 hours. In some embodiments, the methods described herein comprise administering (a) 40 mg of a PRMT5 inhibitor daily, and (b) 1200 mg / m 2 of 5-fluoracil administered to a patient by IV continuous infusion for 44 hours. In some embodiments, the methods described herein comprise administering (a) 80 mg of a PRMT5 inhibitor daily, and (b) 1200 mg / m 2 of 5-fluoracil administered to a patient by IV continuous infusion for 44 hours. In some embodiments, the methods described herein comprise administering (a) 120 mg of a PRMT5 inhibitor daily, and (b) 1200 mg / m 2of 5-fluoracil administered to a patient by IV continuous infusion for 44 hours. In some embodiments, the methods described herein comprise administering (a) 480 mg of a PRMT5 inhibitor daily, and (b) 1200 mg / m 2 of 5-fluoracil administered to a patient by IV continuous infusion for 44 hours. In some embodiments, the methods described herein include administering (a) 800 mg of a PRMT5 inhibitor daily, and (b) 1200 mg / m 2 of 5-fluoracil administered to a patient by IV continuous infusion for 44 hours. In some embodiments, the methods described herein comprise administering (a) 2000 mg of a PRMT5 inhibitor daily, and (b) 1200 mg / m 2 of 5-fluoracil administered to the patient by IV continuous infusion for 44 hours.

[0038] cancer In some embodiments, the cancer is an MTAP-deficient cancer and / or an MTA-accumulated cancer. "MTAP-deficiency-associated," or "MTAP-deficient," or "MTAP-deficient" disease (e.g., a proliferative disorder, e.g., cancer) or "MTAP-associated" disease (e.g., a proliferative disorder, e.g., cancer) or "characterized by MTAP deficiency" disease (e.g., a proliferative disorder, e.g., cancer) and the like refer to a condition (e.g., a proliferative disorder, e.g., cancer) in which MTAP is deficient in a significant number of cells. For example, in an MTAP-deficiency-associated disease, one or more diseased cells may have significantly reduced post-translational modification, production, expression, levels, stability, and / or activity of MTAP. Examples of MTAP-deficiency-associated diseases include, but are not limited to, glioblastoma, malignant peripheral nerve sheath tumor (MPNST), esophageal cancer (e.g., esophageal squamous cell carcinoma or esophageal adenocarcinoma), bladder cancer (e.g., bladder urothelial carcinoma), pancreatic cancer (e.g., pancreatic adenocarcinoma), mesothelioma, melanoma, non-small cell lung cancer (NSCLC, e.g., lung squamous cell carcinoma or lung adenocarcinoma), astrocytoma, undifferentiated pleomorphic sarcoma, diffuse large B-cell lymphoma (DLBCL), leukemia, head and neck cancer, gastric adenocarcinoma, myxofibrosarcoma, bile duct sarcoma, brain cancer, stomach cancer, kidney cancer, breast cancer, endometrial cancer, urinary tract cancer, liver cancer, soft tissue cancer, pleural cancer, and colorectal cancer or sarcoma. In patients with MTAP-deficiency-associated diseases, it is possible that MTAP is defective in some diseased cells (e.g., cancer cells) but not in other diseased cells. Similarly, some diseased cells may accumulate MTA, while others may not. Thus, the present disclosure encompasses treatment methods involving diseases of these or any other tissues, in which the proliferation of MTAP-deficient and / or MTA-accumulating cells can be inhibited by administering a PRMT5 inhibitor. Some cancer cells deficient in MTAP also lack CDKN2A, and these cells exhibit reduced post-translational modification, production, expression, level, stability, and / or activity of the CDKN2A gene or its product. The genes for MTAP and CDKN2A are located in close proximity on chromosome 9p21, with MTAP located approximately 100 kb telomeric to CDKN2A. Many cancer cell types have CDKN2A / MTAP deficiencies (deficiencies of both genes).Thus, in some embodiments, MTAP-deficient cells are also deficient in CDKN2A.

[0039] In some embodiments, the cancer is acute myeloid leukemia, cancer in adolescents, childhood adrenocortical carcinoma, AIDS-related cancers (e.g., lymphoma and Kaposi's sarcoma), anal cancer, appendiceal cancer, astrocytoma, atypical teratoma, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain stem glioma, brain tumor, breast cancer, bronchial tumor, Burkitt's lymphoma, carcinoid tumor, atypical teratoma, embryonal tumor, germ cell tumor, primary lymphoma, cervical cancer, childhood cancer, chordoma, cardiac tumor, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myeloproliferative disorders (CMD), and chronic myeloproliferative disorders (CRD). Sexual diseases, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS), germinoma, CNS cancer, endometrial cancer, ependymoma, esophageal cancer, nasal neuroblastoma, Ewing's sarcoma, extracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, fibrous histiocytoma of bone, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic tumor, hairy cell leukemia, head and neck cancer, cardiac cancer, liver cancer, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumor, pancreatic neuroendocrine tumor, kidney cancer , laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma, metastatic squamous cell neck cancer of unknown primary, midline duct cancer, oral cancer, multiple endocrine neoplasia syndrome, multiple myeloma / plasmacytoma, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, multiple myeloma, Merkel cell carcinoma, malignant mesothelioma, malignant fibrous histiocytoma and osteosarcoma of bone, nasal cavity and paranasal sinus cancer, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer (NSCLC), oral cavity cancer, lip and oral cavity cancer, oropharyngeal cancer, ovarian cancer, The cancer may be pancreatic cancer, papillomatosis, paraganglioma, sinus and nasal cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pleuropulmonary blastoma, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, transitional cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, stomach (gastric) cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, trophoblastic tumor, childhood anomaly cancer, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, or virus-induced cancer. In some cases, the cancer is pancreatic cancer, esophageal cancer, melanoma, lung cancer, mixed Müllerian carcinoma, ovarian cancer, or gallbladder cancer.

[0040] In some embodiments, the cancer is glioblastoma, malignant peripheral nerve sheath tumor (MPNST), esophageal cancer (e.g., esophageal squamous cell carcinoma or esophageal adenocarcinoma), bladder cancer (e.g., bladder urothelial carcinoma), pancreatic cancer (e.g., pancreatic adenocarcinoma), mesothelioma, melanoma, non-small cell lung cancer (NSCLC, e.g., lung squamous cell carcinoma or lung adenocarcinoma), astrocytoma, undifferentiated pleomorphic sarcoma, diffuse large B-cell lymphoma (DLBCL), leukemia, head and neck cancer, gastric adenocarcinoma, myxofibrosarcoma, bile duct sarcoma, brain cancer, stomach cancer, kidney cancer, breast cancer, endometrial cancer, urinary tract cancer, liver cancer, soft tissue cancer, pleural cancer, and colorectal cancer or sarcoma.

[0041] Pharmaceutical Formulations and Routes of Administration Pharmaceutical compositions containing the PRMT5 inhibitors described herein can be manufactured in a conventional manner, for example, by conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilizing processes. Proper formulation depends upon the route of administration chosen.

[0042] Observation of treatment effectiveness The effectiveness of a given cancer treatment can be determined by a skilled clinician. However, if, for example, any one or all of the signs or symptoms of a tumor are changed in a beneficial manner, or other clinically recognized symptoms are improved or even ameliorated by at least 10% after treatment with, for example, a drug as described herein, the treatment is considered to be an "effective treatment," as this term is used herein. Efficacy can also be measured by an individual's failure to worsen (e.g., halting the progression of the disease), as assessed by the need for hospitalization or medical intervention. Methods for measuring these indicators are known to those skilled in the art and / or are described herein.

[0043] The descriptions of embodiments of the present disclosure are not intended to be exhaustive or to limit the disclosure to the precise form disclosed. Specific embodiments and examples of the present disclosure have been described herein for illustrative purposes, but those skilled in the art will recognize that various equivalent modifications are possible within the scope of the present disclosure. The teachings of the present disclosure provided herein may be applied to other procedures or methods, as appropriate. The various embodiments described herein may be combined to provide further embodiments. Where appropriate, aspects of the present disclosure can be modified to employ the compositions, functions, and concepts of the above references and applications to provide still further embodiments of the present disclosure. These and other modifications to the present disclosure can be made in light of the detailed description.

[0044] Specific elements of any of the foregoing embodiments may be combined with or substituted for elements of other embodiments. Furthermore, although advantages associated with certain embodiments of the present disclosure have been described in connection with those embodiments, other embodiments may also exhibit such advantages, and not all embodiments necessarily exhibit such advantages to fall within the scope of the present disclosure.

[0045] All patents and other publications identified are expressly incorporated herein by reference for the purpose of describing and disclosing, for example, the methodology described in such publications that might be used in connection with the present disclosure. These publications are provided solely for their disclosure prior to the filing date of the present application.

[0046] Embodiment 1. A method for treating cancer in a patient in need thereof, comprising administering to the patient an amount of a PRMT5 inhibitor ranging from 40 mg to 2000 mg, wherein the PRMT5 inhibitor is a compound of formula I or compound B: [ka] (In the formula, X 1 is NH, N(C1-C6 alkyl), O or S, X2 is N(C1-C6 alkyl), O or S, Y 2 is H, C1-C6 alkyl or C1-C6 haloalkyl, Z 1 and Z 2 each is independently H, F, or C1-C6 alkyl, and Z 3 , Z 4 , Z 5 and Z 6 each is independently H, C1-C6 alkyl, or chloride. or a pharmaceutically acceptable salt thereof.

[0047] 2. The method of embodiment 1, wherein the cancer is an MTAP-null cancer.

[0048] 3. The method of embodiment 2, wherein the MTAP-null cancer is glioblastoma, mesothelioma, soft tissue sarcoma, esophageal cancer, melanoma, lymphoma / leukemia, head and neck cancer, cholangiocarcinoma, gastric cancer, glioma, thymoma, adenoid cystic carcinoma, pancreatic cancer, lung cancer, breast cancer, liver cancer, or bladder cancer.

[0049] 4. The method of embodiment 3, wherein the lung cancer is non-squamous cell lung cancer (NSCLC).

[0050] 5. The PRMT5 inhibitor has the formula (S)-I: [ka] or a pharmaceutically acceptable salt thereof.

[0051] 6.X 1 is O, Z 1 and Z 2 are H, respectively, X 2 is O, Z 3 , Z 4 , Z 5 and Z 6 are H, respectively, Y 2 is C1-C6 haloalkyl, and Y 2 6. The method of any one of embodiments 1 to 5, wherein is CF3.

[0052] 7. The method of any one of embodiments 1-6, further comprising administering paclitaxel to the subject.

[0053] 8.(a) 40 to 2000 mg of a PRMT5 inhibitor and (b) 75 mg / m 2 Paclitaxel and 8. The method of embodiment 7, comprising administering to a subject

[0054] 9.(a) 40 to 2000 mg of a PRMT5 inhibitor and (b) 100 mg / m 2 Paclitaxel and 8. The method of embodiment 7, comprising administering to a subject

[0055] 10.(a) 40–2000 mg of a PRMT5 inhibitor and (b) 135 mg / m 2 Paclitaxel and 8. The method of embodiment 7, comprising administering to a subject

[0056] 11. The method of any one of embodiments 1-6, further comprising administering carboplatin to the subject.

[0057] 12.(a) 40–2000 mg of a PRMT5 inhibitor and (b) Carboplatin with AUC5-AUC6 12. The method of embodiment 11, comprising administering to a subject

[0058] 13. The method of any one of embodiments 1-6, further comprising administering gemcitabine to the subject.

[0059] 14.(a) 40–2000 mg of a PRMT5 inhibitor and (b) 1000 mg / m 2 ~1250mg / m 2 Gemcitabine and 14. The method of embodiment 13, comprising administering to a subject

[0060] 15. The method of any one of embodiments 1-6, further comprising administering irinotecan to the subject.

[0061] 16.(a) 40–2000 mg of a PRMT5 inhibitor and (b) 150 mg / m 2 ~180mg / m 2 with irinotecan 16. The method of embodiment 15, comprising administering to a subject

[0062] 7. The method of any one of embodiments 1-6, further comprising administering 17.5-fluoracil to the subject.

[0063] 18.(a) 40–2000 mg of a PRMT5 inhibitor and (b) 400 mg / m 2 ~1200mg / m 2 5-Fluoracil and 18. The method of embodiment 17, comprising administering to a subject

[0064] 19. The method of any one of embodiments 1-6, further comprising administering pemetrexed to the subject.

[0065] 20.(a) 40–2000 mg of a PRMT5 inhibitor and (b) 500 mg / m 2 Pemetrexed and 20. The method of embodiment 19, comprising administering to a subject

[0066] 21. The PRMT5 inhibitor is compound G: [ka] 21. The method of any one of embodiments 1 to 20, comprising a compound having the structure:

[0067] 22. The method of any one of embodiments 1-20, wherein the PRMT5 inhibitor comprises compound B or a salt thereof.

[0068] 23. The PRMT5 inhibitor is Compound A: [ka] 21. The method of any one of embodiments 1 to 20, comprising a compound having the structure:

[0069] 24. A method of treating cancer in a patient in need thereof, comprising: (a) a PRMT5 inhibitor in an amount ranging from 40 mg to 2000 mg, the compound being represented by Formula 1 or Compound B [ka] (In the formula, X 1 is NH, N(C1-C6 alkyl), O or S, X 2 is N(C1-C6 alkyl), O or S, Y 2 is H, C1-C6 alkyl or C1-C6 haloalkyl, Z 1 and Z 2 each is independently H, F, or C1-C6 alkyl, and Z 3 , Z 4 , Z 5 and Z 6 each is independently H, C1-C6 alkyl, or chloride. or a pharmaceutically acceptable salt thereof; and (b) Standard of care therapy for the treatment of cancer; to a patient.

[0070] 25. The method of embodiment 24, wherein the standard of care therapy comprises chemotherapy.

[0071] 26. The method of embodiment 25, wherein the chemotherapy comprises paclitaxel, carboplatin, gemcitabine, irinotecan, 5-fluoracil, or pemetrexed, or a combination thereof.

[0072] 27. The PRMT5 inhibitor is represented by the formula (S)-I: [ka] or a pharmaceutically acceptable salt thereof.

[0073] 28.X 1 is O, Z 1 and Z 2 are H, respectively, X 2 is O, Z 3 , Z 4 , Z 5 and Z 6 are H, respectively, Y 2 is C1-C6 haloalkyl, and Y 2 28. The method of any one of embodiments 24-27, wherein is CF3.

[0074] 29. The method of any one of embodiments 25-28, wherein the chemotherapy comprises paclitaxel.

[0075] 30.(a) 40–2000 mg of a PRMT5 inhibitor and (b) 75 mg / m 2 ~135mg / m 2 Paclitaxel and 30. The method of embodiment 29, comprising administering to a subject

[0076] 31. The method of any one of embodiments 25-28, wherein the chemotherapy comprises carboplatin.

[0077] 32.(a) 40–2000 mg of a PRMT5 inhibitor and (b) Carboplatin with AUC5-AUC6 32. The method of embodiment 31, comprising administering to a subject

[0078] 33. The method of any one of embodiments 25-28, wherein the chemotherapy comprises gemcitabine.

[0079] 34.(a) 40–2000 mg of a PRMT5 inhibitor and (b) 1000 mg / m 2 ~1250mg / m 2 Gemcitabine and 34. The method of embodiment 33, comprising administering to a subject

[0080] 35. The method of any one of embodiments 25-28, wherein the chemotherapy comprises irinotecan.

[0081] 36.(a) 40–2000 mg of a PRMT5 inhibitor and (b) 150 mg / m 2 ~180mg / m 2 with irinotecan 36. The method of embodiment 35, comprising administering to a subject

[0082] 37. The method of any one of embodiments 25-28, wherein the chemotherapy comprises 5-fluoracil.

[0083] 38.(a) 40–800 mg of a PRMT5 inhibitor and (b) 400 mg / m 2 ~1200mg / m 2 5-Fluoracil and 38. The method of embodiment 37, comprising administering to a subject

[0084] 39. The method of any one of embodiments 25-28, wherein the chemotherapy comprises pemetrexed.

[0085] 40.(a) 40–2000 mg of a PRMT5 inhibitor and (b) 500 mg / m 2 Pemetrexed and 40. The method of embodiment 39, comprising administering to a subject

[0086] 41. The PRMT5 inhibitor is compound G: [ka] or a salt thereof.

[0087] 42. The method of any one of embodiments 24-40, wherein the PRMT5 inhibitor comprises compound B or a salt thereof.

[0088] 43. The PRMT5 inhibitor is Compound A: [ka] or a salt thereof.

[0089] 44. The method of any one of embodiments 24-43, wherein the PRMT5 inhibitor and the standard of care therapy are administered simultaneously.

[0090] 45. The method of any one of embodiments 24-43, wherein the PRMT5 inhibitor and the standard of care therapy are administered sequentially. [Example]

[0091] Example 1 - Combination of a Methylthioadenosine (MTA)-Cooperative PRMT5 Inhibitor with Chemotherapy in Patients with Advanced Methylthioadenosine Phosphorylase (MTAP)-Null Solid Tumors Compound G is an MTA-cooperative PRMT5i that preferentially targets MTA-bound PRMT5, which is abundant in MTAP-null tumors, and thus represents a novel strategy to improve the therapeutic margin of this class of inhibitors.

[0092] Eligible patients (≥18 years of age) with histologically confirmed locally advanced / metastatic ST not amenable to surgery and / or radiation, homozygous MTAP and / or CDKN2A deletion (by local next-generation sequencing), loss of MTAP protein in ST (by central immunohistochemistry), measurable disease, ECOG PS 0-1, adequate hematopoietic, renal, hepatic, pulmonary, cardiac, and coagulation function, and glycemic control were included. This study consists of three parts, each with its own subparts. Here, we describe Parts 1c-h (dose expansion of Compound G) and Part 2 (dose-finding [a] and dose expansion [b] of Compound G plus docetaxel) in patients with squamous non-small cell lung cancer (NSCLC) (1c), adenocarcinoma (1d), cholangiocarcinoma (1e), head and neck squamous cell carcinoma (1f), pancreatic adenocarcinoma (1g), other ST and lymphoma excluding primary brain tumors (1h), and NSCLC (2a / b). Primary endpoints include dose-limiting toxicity, adverse events, ECG, laboratory abnormalities, and vital signs. Secondary endpoints include Cmax, Tmax, and AUC after single or multiple doses, time to response, stable disease, progression-free survival, overall survival, objective response, disease control, and duration of response. The study will enroll approximately 290 and 50 patients in Part 1 and Part 2, respectively.

[0093] The above clinical trial is repeated using compound (G) in combination with some other chemotherapeutic agent (eg, paclitaxel, carboplatin, gemcitabine, irinotecan, 5-fluoracil, or pemetrexed).

[0094] Example 2 – Efficacy of PRMT5 inhibitors in MTAP-null cell lines In the following examples, the efficacy of PRMT5 inhibitors (e.g., Compound B and Compound G) was evaluated in MTAP-null (HTAP and H116) cell lines. Compound A was evaluated using the same protocol.

[0095] Cells were plated at optimal cell density in 90 μL of growth medium into 96-well black-walled, clear-bottom tissue culture plates. The cells were incubated at room temperature for 30 minutes before being placed in an incubator at 37°C and 5% CO2. The following day, PRNT5 inhibitors (e.g., Compound B and Compound G) were serially diluted in DMSO at a 1:3 dilution factor into 96-well V-bottom plates. Secondary compound dilutions (1:100) in medium were performed in 96-well V-bottom plates by adding 3 μL of the primary dilution to 297 μL of culture medium. Finally, 10 μL of the secondary compound dilution was added to cells in triplicate (1:10 dilution; final DMSO concentration 0.1%). After addition, the plates were incubated at 37°C and 5% CO2. Cell viability was measured by CellTiter-Glo luminescence assay on day 6 after treatment. Percentage of control (POC) values ​​were calculated as follows: POC = 100 * (treatment / vehicle). The mean POC value was then calculated for each treatment condition and used to fit dose-response curves in GraphPad Prism 7 using a four-parameter logistic curve fit.

[0096] Global SDMA levels in HAP1 WT and MTAP-null cells were assessed by ELISA assay on day 3 after treatment, and the results are shown in Figure 1C.

[0097] Global SDMA levels in HCT116 WT and MTAP-null cells were assessed by in-cell imaging assay on day 4 after treatment, and the results are shown in Figure 1D.

[0098] After treatment with Compound B, SDMA levels were lower in both HAP1 and HCT116 MTAP-null cells (IC50 = 0.0002 μM) compared to HCT116 MTAP-WT cells (IC50 = 0.050 μM). See Figures 1C and 1D. Compound B also selectively inhibited the proliferation of both HAP1 and HCT116 MTAP-null cells (IC50 = 0.027 μM) compared to HCT116 MTAP-WT cells (IC50 = 0.63 μM). See Figures 1A and 1B. Profiling Compound B in an expanded panel of tumor cell lines demonstrated that Compound B inhibited the proliferation of most MTAP-null cells while having minimal effects on MTAP-WT cells. Finally, in vitro mechanism-of-action studies demonstrated that treatment with Compound B induced DNA damage, as indicated by increased H2AX phosphorylation and G2 / M cell cycle arrest in MTAP-null cells (data not shown). In vivo, oral administration of Compound B selectively inhibited SDMA and tumor growth in HCT116 MTAP-null tumor xenografts compared with HCT116 MTAP-WT xenografts. See Figure 2.

[0099] Example 3 – Combination of PRMT5 inhibitors with paclitaxel in NSCLC cell lines NSCLC cell lines (H292 A549) were treated with a combination of a PRMT5 inhibitor (i.e., Compound B or Compound G) and paclitaxel for 6 days. A 1.9-fold dilution series was performed for the PRMT5 inhibitor (i.e., Compound B or Compound G), and a 1.2- to 1.7-fold dilution series was performed for the combination, creating an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured by the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fractional activity (Fa) using the following formula:

number

[0100] Synergy analysis was performed using CalcuSyn software, and CI scores were determined based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 1-4. *CI values ​​(Calcusyn): Strong synergy: 0.1-0.3; Synergy: 0.3-0.7; Moderate synergy: 0.7-0.85; Slight synergy: 0.85-0.9; Near-additive: 0.9-1.1.

[0101] [Table 1]

[0102] [Table 2]

[0103] [Table 3]

[0104] [Table 4]

[0105] As shown in Tables 1-4, the majority of CI scores fall within the range of moderate synergy and slight synergy.

[0106] Example 4 – Combination of PRMT5 inhibitors with paclitaxel in NSCLC cell lines NSCLC cell lines (H292, A549) were treated with a combination of a PRMT5 inhibitor (i.e., Compound A) and paclitaxel for 6 days. A 1.9-fold dilution series was performed for the PRMT5 inhibitor (Compound A), and a 1.2- to 1.7-fold dilution series was performed for the combination, creating an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescent assay.

[0107] Example 5 – Combination of PRMT5 inhibitors with gemcitabine in pancreatic cell lines Pancreatic cancer cell lines (MIAPACA2T2, PSN1) were treated with a combination of a PRMT5 inhibitor (i.e., Compound B or Compound G) and gemcitabine for 6 days. A 1.9-fold dilution series was performed for the PRMT5 inhibitor (i.e., Compound B or Compound G), and a 1.2- to 1.7-fold dilution series was performed for the combination, creating an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured by the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fractional activity (Fa) using the following formula:

number

[0108] Synergy analysis was performed using CalcuSyn software, and CI scores were determined based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 5-8. *CI values ​​(Calcusyn): Strong synergy: 0.1-0.3; Synergy: 0.3-0.7; Moderate synergy: 0.7-0.85; Slight synergy: 0.85-0.9; Near-additive: 0.9-1.1.

[0109] [Table 5]

[0110] [Table 6]

[0111] [Table 7]

[0112] [Table 8]

[0113] As shown in Tables 5-8, the majority of CI scores are in the range of moderate synergy.

[0114] Example 6 – Combination of PRMT5 inhibitors with gemcitabine in pancreatic cell lines Pancreatic cancer cell lines (MIAPACA2T2, PSN1) were treated with a combination of a PRMT5 inhibitor (i.e., Compound A) and gemcitabine for 6 days. A 1.9-fold dilution series was performed for the PRMT5 inhibitor (i.e., Compound A), and a 1.2- to 1.7-fold dilution series was performed for the combination, creating an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescent assay.

[0115] Example 7 – Combination of PRMT5 inhibitors with carboplatin in pancreatic cell lines Pancreatic cancer cell lines (MIAPACA2T2, PSN1) were treated with a combination of a PRMT5 inhibitor (i.e., Compound B or Compound G) and carboplatin for 6 days. A 1.9-fold dilution series was performed for the PRMT5 inhibitor (i.e., Compound B or Compound G), and a 1.2- to 1.7-fold dilution series was performed for the combination, creating an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured by the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fractional activity (Fa) using the following formula:

number

[0116] Synergy analysis was performed using CalcuSyn software, and CI scores were determined based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 9-12. *CI values ​​(Calcusyn): Strong synergy: 0.1-0.3; Synergy: 0.3-0.7; Moderate synergy: 0.7-0.85; Slight synergy: 0.85-0.9; Near-additive: 0.9-1.1.

[0117] [Table 9]

[0118] [Table 10]

[0119] [Table 11]

[0120] [Table 12]

[0121] As shown in Tables 10-12, the majority of CI scores are within the synergistic range.

[0122] Example 8 – Combination of PRMT5 inhibitors with carboplatin in pancreatic cell lines Pancreatic cancer cell lines (MIAPACA2T2, PSN1) were treated with a combination of a PRMT5 inhibitor (i.e., Compound A) and carboplatin for 6 days. A 1.9-fold dilution series was performed for the PRMT5 inhibitor (i.e., Compound A), and a 1.2- to 1.7-fold dilution series was performed for the combination, creating an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescent assay.

[0123] Example 9 – Combination of PRMT5 inhibitors with pemetrexed in NSCLC cell lines NSCLC cancer cell lines (H292, A549) were treated with a combination of a PRMT5 inhibitor (i.e., Compound B or Compound G) and pemetrexed for 6 days. A 1.9-fold dilution series was performed for the PRMT5 inhibitor (i.e., Compound B or Compound G), and a 1.2- to 1.7-fold dilution series was performed for the combination, creating an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured by the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fractional activity (Fa) using the following formula:

number

[0124] Synergy analysis was performed using CalcuSyn software, and CI scores were determined based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 13-16. *CI values ​​(Calcusyn): strong synergy: 0.1-0.3; synergy: 0.3-0.7; moderate synergy: 0.7-0.85; slight synergy: 0.85-0.9; nearly additive: 0.9-1.1.

[0125] [Table 13]

[0126] [Table 14]

[0127] [Table 15]

[0128] [Table 16]

[0129] As shown in Tables 13-16, the majority of CI scores are within the synergistic range.

[0130] Example 10 – Combination of PRMT5 inhibitors with pemetrexed in NSCLC cell lines NSCLC cancer cell line (H292) was treated with a combination of a PRMT5 inhibitor (i.e., Compound A) and pemetrexed for 6 days. A 1.9-fold dilution series was performed for the PRMT5 inhibitor (i.e., Compound A), and a 1.2- to 1.7-fold dilution series was performed for the combination, creating an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescent assay.

[0131] Example 11 – Combination of PRMT5 inhibitors with irinotecan in pancreatic cell lines Pancreatic cancer cell lines (MIAPACA2T2, PSN1) were treated with a combination of a PRMT5 inhibitor (i.e., Compound B or Compound G) and irinotecan for 6 days. A 1.9-fold dilution series was performed for the PRMT5 inhibitor (i.e., Compound B or Compound G), and a 1.2- to 1.7-fold dilution series was performed for the combination, creating an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured by the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fractional activity (Fa) using the following formula:

number

[0132] Synergy analysis was performed using CalcuSyn software, and CI scores were determined based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 17-20. *CI values ​​(Calcusyn): Strong synergy: 0.1-0.3; Synergy: 0.3-0.7; Moderate synergy: 0.7-0.85; Slight synergy: 0.85-0.9; Near-additive: 0.9-1.1.

[0133] [Table 17]

[0134] [Table 18]

[0135] [Table 19]

[0136] [Table 20]

[0137] As shown in Tables 17-20, the majority of CI scores are in the synergy range for MIAPACA2T2 and in the moderate synergy range for PSN1.

[0138] Example 12 – Combination of PRMT5 inhibitors with irinotecan in pancreatic cell lines Pancreatic cancer cell lines (MIAPACA2T2, PSN1) were treated with a combination of a PRMT5 inhibitor (i.e., Compound A) and irinotecan for 6 days. A 1.9-fold dilution series was performed for the PRMT5 inhibitor (i.e., Compound A), and a 1.2- to 1.7-fold dilution series was performed for the combination, creating an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescent assay.

[0139] Example 13 – Combination of PRMT5 inhibitors with 5-fluoracil (5-FU) in pancreatic cell lines Pancreatic cancer cell lines (MIAPACA2T2, PSN1) were treated with a combination of PRMT5 inhibitors (i.e., Compound B or Compound G) and 5-FU for 6 days. A 1.9-fold dilution series was performed for the PRMT5 inhibitors (i.e., Compound B or Compound G), and a 1.2- to 1.7-fold dilution series was performed for the combination, creating an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured by the CellTiter-Glo luminescence assay. Raw luminescence values ​​were converted to fractional activity (Fa) using the following formula:

number

[0140] Synergy analysis was performed using CalcuSyn software, and CI scores were determined based on the drug concentrations used and the corresponding Fa values. The results are shown in Tables 21-24. *CI values ​​(Calcusyn): strong synergy: 0.1-0.3; synergy: 0.3-0.7; moderate synergy: 0.7-0.85; slight synergy: 0.85-0.9; nearly additive: 0.9-1.1.

[0141] [Table 21]

[0142] [Table 22]

[0143] [Table 23]

[0144] [Table 24]

[0145] As shown in Tables 21-24, the majority of CI scores are within the synergistic range for MIAPACA2T2 and within the additive range for PSN1.

[0146] Example 14 – Combination of PRMT5 inhibitors with 5-fluoracil (5-FU) in pancreatic cell lines Pancreatic cancer cell lines (MIAPACA2T2, PSN1) were treated with a combination of a PRMT5 inhibitor (i.e., Compound A) and 5-FU for 6 days. A 1.9-fold dilution series was performed for the PRMT5 inhibitor (i.e., Compound A), and a 1.2- to 1.7-fold dilution series was performed for the combination, creating an 8 x 10 dose matrix, including a DMSO-only control. Cell viability was measured using the CellTiter-Glo luminescent assay.

[0147] Example 15 – Growth in multiple MTAP-null tumor xenograft models inhibited by PRMT5 inhibitors Six female NOD / SCID mice were implanted with patient-derived tumor xenograft (PDX) models of pancreatic cancer, ovarian cancer, esophageal cancer, melanoma, lung cancer, brain cancer, mixed Müllerian duct carcinoma, or gallbladder cancer. The mean tumor volume in each group was 100–200 mm. 3Mice were assigned to two different test groups according to tumor volume and started on oral administration of 100 mg / kg of vehicle or Compound B once daily. The plotted data represent TGI (tumor growth inhibition), n=3 in each group.

[0148] Furthermore, treatment with Compound B inhibits the growth of multiple MTAP-null tumor xenograft models, BXPC3 (PDAC) and DOHH2 (DLBCL) (Figure 3). When Compound B was profiled against a panel of over 20 PDX models (Figure 6), greater than 50% tumor growth inhibition was observed in the majority of PDX models with MTAP gene deletion (Figure 4).

[0149] The data provided herein demonstrate that PRMT5 inhibitors that selectively target PRMT5 in concert with MTA may represent a novel and compelling therapeutic approach for the treatment of MTAP-null cancers.

[0150] Example 16 – Growth in multiple MTAP-null tumor xenograft models inhibited by PRMT5 inhibitors Female NOD / SCID mice were implanted with patient-derived tumor xenograft (PDX) models of pancreatic, ovarian, esophageal, melanoma, lung, brain, mixed Müllerian, or gallbladder cancer. Mice were assigned to two different test groups based on tumor volume and initiated with oral daily dosing of 100 mg / kg of vehicle or Compound A. Tumor volume was assessed and plotted over time to demonstrate tumor growth inhibition (TGI).

[0151] Example 17 - Tumor volume in NSCLC animal models suppressed by a combination of a PRMT5 inhibitor and paclitaxel Ten female NOD / SCID mice were implanted with H292 NSCLC tumor xenografts. The mean tumor volume in each group was 100–200 mm. 3Mice were assigned to two different test groups according to tumor volume and started to receive oral administration of vehicle or compound G (100 mg / kg) or compound B (100 mg / kg) in combination with paclitaxel (20 mg / kg) once a day. The plotted data represent TGI (tumor growth inhibition), n=10 in each group.

[0152] The results showed that the combination of Compound G with paclitaxel resulted in significant antitumor activity compared to single agents alone in H292 NSCLC xenografts. See Figure 7.

[0153] Example 18 - Tumor volume in NSCLC animal models suppressed by a combination of a PRMT5 inhibitor and paclitaxel Female NOD / SCID mice were implanted with H292 NSCLC tumor xenografts. Mice were assigned to two different test groups based on tumor volume, and were administered once daily orally with vehicle or Compound A (100 mg / kg) in combination with paclitaxel (20 mg / kg). Tumor volume was assessed and plotted over time to indicate tumor growth inhibition (TGI).

[0154] Example 19 – Tumor volume in H292 NSCLC animal model suppressed by combination of PRMT5 inhibitor and paclitaxel Ten female NOD / SCID mice were implanted with H292 tumor xenografts. The mean tumor volume in each group was 100–200 mm. 3 Mice were assigned to two different test groups according to tumor volume and started on oral administration of vehicle or Compound G (100 mg / kg) in combination with paclitaxel (20 mg / kg) once daily. Paclitaxel was administered intraperitoneally (IP) starting on day 10, and then every other day for a total of five doses. Data plotted represent TGI (tumor growth inhibition), n=10 in each group.

[0155] The results showed that the combination of Compound G and paclitaxel resulted in significant antitumor activity compared to single-agent H292 NSCLC xenografts. See Figure 7. Similarly, the combination of Compound B and paclitaxel resulted in significant antitumor activity compared to single-agent H292 NSCLC xenografts. See Figure 8.

[0156] Example 20 – Tumor volume in H292 NSCLC animal model suppressed by combination of PRMT5 inhibitor and paclitaxel Female NOD / SCID mice are implanted with H292 xenografts. Mice are assigned to two different test groups according to tumor volume, and begin oral administration of vehicle or Compound A (100 mg / kg) in combination with paclitaxel once daily. Tumor volume is assessed over time and plotted to show tumor growth inhibition (TGI).

[0157] Example 21 – PRMT5 inhibitor monotherapy in adult patients with metastatic or locally advanced MTAP-null solid tumors The following studies will be conducted to evaluate the safety and tolerability of PRMT5 inhibitor monotherapy in adult patients with metastatic or locally advanced MTAP-null solid tumors and to determine the maximum tolerated dose (MTD) or recommended phase 2 dose (RP2D). The pharmacokinetics (PK) of PRMT5 inhibitor monotherapy will be evaluated. Additionally, the following will be evaluated: objective response rate (ORR), disease control rate (DCR), duration of response (DoR), time to response (TTR), stable disease (SD), progression-free survival (PFS), and overall survival (OS) of PRMT5 inhibitors in adult patients with MTAP-null solid tumors.

[0158] The study will be conducted in three parts, each with subparts: Part 1a / b (dose-finding, five dose levels) will enroll approximately 30 patients from all eligible tumor types.

[0159] Treatment will continue until progression or withdrawal. Safety follow-up will occur approximately 30 (± 3) days after the last dose of PRMT5 inhibitor or before initiating other therapy, whichever occurs first. Long-term follow-up will occur every 6 months for up to 2 years after the first dose for all patients who have not withdrawn consent.

[0160] Intrapatient dose escalation is permitted. Patients who complete the dose-limiting toxicity (DLT) period may proceed to higher dose levels not exceeding the highest dose level deemed safe by the Dose Level Review Team (DLRT), provided that no DLTs are reported for the patient during or after the DLT period and the patient does not experience any adverse events of grade 2 or higher on treatment (deemed by the investigator to be treatment-related).

[0161] The dose-finding study will estimate the MTD using a Bayesian logistic regression model (BLRM) design. Upon completion of each cohort, the DLRT will recommend the following dose level: (1) the recommended dose level from the BLRM and a recommended dose level based on evaluation of available safety data, laboratory results, and PK information; (2) the RP2D may be identified based on new safety, efficacy, PK, and PD data prior to reaching the MTD.

[0162] Before proceeding to dose expansion, this dose-finding part of the study requires a minimum of six DLT-evaluable patients to be treated at the RP2D as monotherapy.

[0163] [Table 25]

[0164] Evaluation item definition - Objective response (defined as best overall response of confirmed complete or partial response per RECIST v1.1) as determined using investigator tumor assessment.

[0165] - Disease control is defined as confirmed objective response or stable disease (SD) per RECIST v1.1.

[0166] Duration of response (DoR) is defined as the time from first documented and subsequently confirmed objective response per RECIST v1.1 to first documented disease progression or death from any cause, whichever occurs first.

[0167] - Time to response is defined as the time from enrollment to the first documented and subsequently confirmed objective response per RECIST v1.1.

[0168] - Duration of SD is defined as the time from first dose of PRMT5 inhibitor to first documented radiographic disease progression or death from any cause (whichever occurs first) per RECIST v1.1.

[0169] - Progression-free survival (PFS) is defined as the time from first dose of PRMT5 inhibitor to first documented radiological disease progression or death from any cause (whichever occurs first in the absence of subsequent anticancer therapy). PFS is censored at the last evaluable post-baseline tumor assessment before subsequent anticancer therapy, or at the first dose of PRMT5 inhibitor if there is no subsequent anticancer therapy. Progression is based on RECIST v1.1 derived using investigator tumor assessment.

[0170] Overall survival (OS) is defined as the time from the first dose of a PRMT5 inhibitor to death from any cause. OS is censored at the last known day of survival up to the data cutoff date.

[0171] Example 22 – Combination of PRMT5 inhibitors with paclitaxel or carboplatin in NSCLC cell lines NSCLC cell line (H292) was treated for 6 days with a combination of a PRMT5 inhibitor (i.e., Compound B) and either paclitaxel or carboplatin. The PRMT5 inhibitor (Compound B) was diluted 1.9-fold, and the combination was diluted 1.2-1.7-fold, creating an 8 x 10 dose matrix, including a DMSO-only control.

[0172] Synergy analysis was performed using CalcuSyn software to determine combination index (CI) scores based on the drug concentrations used and the corresponding Fa values. CI<1 indicates synergy; CI=1 indicates additive; CI>1 indicates antagonism. The results are shown in Figures 12 and 13.

[0173] To assess cell proliferation after combination treatment, cell nuclei were counted over 10 days using an IncuCyte live cell imager. Results showed that the combination of Compound B with paclitaxel (FIG. 14A) and Compound B with carboplatin (FIG. 14B) resulted in significant antitumor cell proliferation activity compared to paclitaxel or carboplatin alone in NSCLC (H292) cell lines.

[0174] Example 23 – Tumor volume in H292 MTAP-null NSCLC xenografts suppressed by the combination of a PRMT5 inhibitor with paclitaxel or carboplatin Ten female NOD / SCID mice were implanted with H292 tumor xenografts. The mean tumor volume in each group was 100–200 mm. 3 Mice were assigned to two different study groups based on tumor volume and started on oral daily treatment with vehicle or B1 (100 mg / kg) in combination with paclitaxel (20 mg / kg) or carboplatin (60 mg / kg). Paclitaxel was administered intraperitoneally (IP) starting on day 10, then every other day for a total of five doses. Carboplatin was administered intraperitoneally (IP) starting on day 10, then once a week for a total of three doses. Data plotted represent TGI (tumor growth inhibition), n=10 per group.

[0175] The results showed that the combination of Compound B with paclitaxel (Figure 15A) and Compound B with carboplatin (Figure 15B) resulted in significant antitumor activity compared to paclitaxel or carboplatin alone in H292 NSCLC xenografts.

Claims

1. A pharmaceutical composition for use in a method of treating cancer in a patient in need thereof, wherein the method comprises administering to the patient an amount of a PRMT5 inhibitor in the range of 40 mg to 2000 mg, the pharmaceutical composition comprises a PRMT5 inhibitor, the PRMT5 inhibitor is compound (G): 【Chemical 1】 A pharmaceutical composition having the structure or a pharmaceutically acceptable salt thereof.

2. The pharmaceutical composition according to claim 1, comprising administering the PRMT5 inhibitor in an amount in the range of 40 mg to 800 mg.

3. The pharmaceutical composition according to claim 1, wherein the cancer is MTAP-null cancer.

4. The pharmaceutical composition according to claim 3, wherein the MTAP-null cancer is glioblastoma, mesothelioma, soft tissue sarcoma, esophageal cancer, melanoma, lymphoma / leukemia, head and neck cancer, cholangiocarcinoma, gastric cancer, glioma, thymoma, adenoid cystic carcinoma, pancreatic cancer, lung cancer, breast cancer, liver cancer or bladder cancer.

5. The pharmaceutical composition according to claim 4, wherein the MTAP-null cancer is lung cancer.

6. The pharmaceutical composition according to claim 5, wherein the lung cancer is non-small cell lung cancer (NSCLC).

7. The pharmaceutical composition according to claim 4, wherein the MTAP-null cancer is pancreatic cancer.

8. The pharmaceutical composition according to claim 7, wherein the pancreatic cancer is pancreatic adenocarcinoma.

9. The pharmaceutical composition according to claim 4, wherein the MTAP-null cancer is esophageal cancer.

10. The pharmaceutical composition according to claim 9, wherein the esophageal cancer is esophageal squamous cell carcinoma or esophageal adenocarcinoma.

11. The pharmaceutical composition according to any one of claims 1 to 10, further comprising administering a standard treatment therapy for treating cancer.

12. The pharmaceutical composition according to claim 11, wherein the standard treatment therapy comprises chemotherapy, and the chemotherapy comprises administration of paclitaxel, carboplatin, gemcitabine, irinotecan, 5-fluorouracil or pemetrexed or a combination thereof.

13. The pharmaceutical composition according to claim 12, wherein the standard treatment therapy comprises administration of paclitaxel.

14. wherein the standard treatment therapy comprises administration of paclitaxel at 75 mg / m ２ to 135 mg / m ２ The pharmaceutical composition according to claim 13.

15. The pharmaceutical composition according to claim 12, wherein the standard treatment therapy comprises administration of carboplatin.

16. The pharmaceutical composition according to claim 15, wherein the standard treatment therapy comprises administration of carboplatin with an AUC of 5 to 6.

17. The pharmaceutical composition according to claim 12, wherein the standard treatment therapy comprises administration of gemcitabine.

18. The standard treatment method is gemcitabine administration at 1000 mg / m ２ to 1250 mg / m ２ The pharmaceutical composition according to claim 17, which comprises gemcitabine administration at 1000 mg / m to 1250 mg / m.

19. The pharmaceutical composition according to claim 12, wherein the standard treatment therapy includes administration of irinotecan.

20. The standard treatment method is 150 mg / m ２ to 180 mg / m ２ The pharmaceutical composition according to claim 19, comprising administration of irinotecan of.

21. The pharmaceutical composition according to claim 12, wherein the standard treatment therapy includes administration of 5-fluorouracil.

22. wherein the standard treatment therapy comprises administration of 5-fluorouracil at 400 mg / m ２ to 1200 mg / m ２ The pharmaceutical composition according to claim 21.

23. The pharmaceutical composition according to claim 12, wherein the standard treatment therapy includes administration of pemetrexed.

24. The standard treatment method includes administration of pemetrexed at 500 mg / m ２ The pharmaceutical composition according to claim 23, wherein the standard treatment method includes administration of pemetrexed at 500 mg / m

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