Combination therapy using a PRMT5 inhibitor and a BCL-2 family inhibitor for cancer treatment

Abstract

Disclosed herein is a method for treating cancer. More specifically, the present disclosure provides a method for treating cancer in a subject using a compound that is a PRMT5 inhibitor, particularly in combination with a BCL-2 family inhibitor.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority from U.S. Provisional Application No. 63 / 403,201, filed September 1, 2022, and U.S. Provisional Application No. 63 / 497,022, filed April 19, 2023, the disclosures of each of which are incorporated herein by reference in their entireties.

[0002] The present disclosure relates to methods of treating cancer. The present disclosure further relates to methods of treating cancer in a subject using a compound that is an inhibitor of protein arginine N-methyltransferase 5 (PRMT5), in combination with a BCL-2 family inhibitor, particularly in combination with a BCL-2 family inhibitor that has activity against BCL-xL.

[0003] Description of Related Art PRMT5 is a type II arginine methyltransferase that catalyzes the transfer of a methyl group from S-adenosyl-L-methionine (SAM) to the omega-nitrogen of the guanidino function of L-arginine residues in proteins (omega-monomethylation) and a second methyl group transfer to the other omega-nitrogen to generate symmetric dimethylarginine (sDMA). PRMT5 forms a complex with methylosome protein 50 (MEP50), which is required for substrate recognition and orientation and for PRMT5-dependent SDMA modification of histone 2A and histone 4 (see, e.g., Ho et al. (2013) PLoS ONE 8(2):e57008).

[0004] Homozygous deletions of p16 / CDKN2a are the most frequent mutations observed in cancer, and these mutations are commonly associated with co-deletions of neighboring genes, including the gene encoding methylthioadenosine phosphorylase (MTAP). It is estimated that approximately 15% of all human cancers harbor homozygous deletions of the MTAP gene (see, e.g., Firestone & Schramm (2017) J. Am. Chem Soc. 139(39):13754-13760).

[0005] Cells lacking MTAP activity have elevated concentrations of methylthioadenosine (MTA), an MTAP substrate that is a potent inhibitor of PRMT5. Elevated MTA inhibits PRMT5 activity, reducing its methylation activity and making cell proliferation more sensitive to further PRMT5 depletion or inhibition. Thus, loss of MTAP activity reduces the methylation activity of PRMT5 and renders cells selectively dependent on PRMT5 activity.

[0006] Despite PRMT5's importance for cell viability and its abundance in cancer, no effective therapeutics have been found to inhibit PRMT5, and there remains a need to develop new PRMT5 inhibitor therapeutics to treat a wide range of cancers.

[0007] BCL-2 family inhibitors are being investigated as cancer treatments. They are believed to work by inhibiting pro-survival signaling mediated by select BCL-2 family members. Inhibition of BCL-2 family members is expected to inhibit cancer cell survival and provide clinical benefit to cancer patients. The Bcl-2 inhibitor venetoclax has been approved for the treatment of chronic lymphocytic leukemia. Navitoclax, an inhibitor of Bcl-2, Bcl-xL, and Bcl-w, is currently in clinical trials for a number of liquid and solid tumors. Navitoclax is being studied as a single agent and in combination with other treatments, including chemotherapy. DT2216, a Bcl-xL inhibitor, is being investigated as a treatment for a variety of solid and liquid tumors. BCL-2 family inhibitors may have different utility for the treatment of various cancer types, depending on their sensitivity to inhibition of one or more BCL-2 family members.

[0008] Despite the importance of BCL-2 family members for cancer cell survival and their frequent role in resistance to cancer therapy, no effective therapies inhibiting BCL-2 family members have been identified. Thus, there remains a need to improve the efficacy of cancer therapies that involve the administration of BCL-2 family inhibitors.

[0009] (Summary of the Invention) The present disclosure provides a method for treating cancer in a subject. Such a method includes administering to the subject a therapeutically effective amount of a PRMT5 inhibitor in combination with a therapeutically effective amount of a BCL-2 family inhibitor. The BCL-2 family inhibitor in such a combination and / or combined therapy may be selected from one or more of the following: ABT-199 (venetoclax), ABT-263 (navitoclax), A-1155463, A-1331852, obatoclax (GX15-070), ABT-737, TW-37, gossypol or (R)-(-)-gossypol, HA14-1, sabutoclax, DT2216, AMG176, PRT1419, AZD5991, S64315 / MIK665, or a combination thereof. Additional BCL-2 family inhibitors are currently being developed and may be used in the embodiments of the invention described herein. The PRMT5 inhibitor may be any of the PRMT5 inhibitors disclosed in WO 2021 / 050915 A1 (e.g., MRTX-1719).

[0010] Also included herein are methods for treating cancer in a subject identified as needing such treatment, including determining whether the subject's cancer is a cancer associated with MTAP homozygous deletion (e.g., an MTAP-associated cancer).

[0011] These and other features and advantages of the present invention will be more fully understood from the following detailed description taken in conjunction with the appended claims, which should be noted that the claims are defined by the description in the specification, rather than by the specific discussion of the features and advantages set forth herein.

[0012] The accompanying drawings are included to provide a further understanding of the methods of the present disclosure, and are incorporated in and constitute a part of this specification. The drawings illustrate one or more embodiments of the present disclosure, and together with the description, serve to explain the principles and operation of the present disclosure. [Brief explanation of the drawings]

[0013] [Figure 1] Figure 1 shows the results of Example 1, in which MRTX1719 (100 mg / kg PO, QD), the BCL-2 selective inhibitor venetoclax (ABT-199) (100 mg / kg PO, QD), or a combination thereof; or MRTX1719 (100 mg / kg PO, QD), the BCL-2 / BCL-xL inhibitor navitoclax (ABT-263) (100 mg / kg PO, QD), or a combination thereof were administered to mice (n=5 / cohort) bearing LU99 xenograft tumors. Data are presented as mean tumor volume + / - SEM. [Figure 2] Figure 2 shows the results of Example 2, in which MRTX1719 (100 mg / kg PO, QD), the BCL-2 / Bcl-w / BCL-xL inhibitor navitoclax (ABT-263) (100 mg / kg PO, QD), or a combination thereof; or MRTX1719 (100 mg / kg PO, QD), the BCL-xL-selective inhibitor A-1331852 (25 mg / kg PO, QD), or a combination thereof were administered to mice bearing NCI-H1437 xenograft tumors (n=4 / cohort). Data are presented as mean tumor volume + / - SEM. [Figure 3] Figure 3 shows the results of Example 3, in which MRTX1719 (100 mg / kg PO, QD), the BCL-2 / Bcl-w / BCL-xL inhibitor navitoclax (ABT-263) (100 mg / kg PO, QD), or a combination thereof; or MRTX1719 (100 mg / kg PO, QD), the BCL-xL-selective inhibitor A-1331852 (25 mg / kg PO, QD), or a combination thereof were administered to mice bearing SW1573 xenograft tumors (n=4 / cohort). Data are presented as mean tumor volume + / - SEM. [Figure 4]Figure 4 shows the results of Example 4, in which MRTX1719 (100 mg / kg PO, QD), the BCL-xL-selective inhibitor A-1331852 (25 mg / kg PO, BID), or a combination thereof was administered to mice (n=5 / cohort) bearing LU99 xenograft tumors. Data are presented as mean tumor volume + / - SEM. Detailed Description of the Invention

[0014] Before the disclosed processes and materials are described, it is to be understood that the aspects described herein are not limited to particular embodiments, which may, of course, vary, and that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting, unless specifically defined herein.

[0015] In view of the present disclosure, the methods and compositions described herein can be configured by those skilled in the art to meet desired needs.The present disclosure provides improvements in the treatment of cancer in subjects.As used herein, the terms "subject" or "patient" are used interchangeably and refer to any animal, including mammals, and most preferably humans.

[0016] The PRMT5 inhibitors disclosed herein exhibit selective activity in MTAP-deficient cancers by binding to and inhibiting PRMT5 when bound to the intracellular metabolite MTA. As described above, MTAP is an enzyme in the methionine salvage pathway, and its deletion in cancer cells leads to the accumulation of MTA in these cancer cells. PRMT5 is an essential enzyme required for cell survival, and therefore, the PRMT5 inhibitors disclosed herein represent a novel approach to selectively treat MTAP-deficient cancers.

[0017] A single mutation rarely causes cancer. The present inventors have found that the treatment of certain cancers with PRMT5 inhibitors can be improved by using combination therapy. In particular, the present inventors have surprisingly found that MTA cooperative PRMT5 inhibitors and BCL-2 family inhibitors (e.g., ABT-199 (venetoclax), ABT-263 (navitoclax), A-1155463, A-1331852, obatoclax (GX15-070), ABT-737, TW-37, gossypol or (R)-(-)-gossypol, HA14-1, sabutoclax, and / or DT2216) inhibit tumor cell viability in vitro in a synergistic manner and provide more potent tumor growth inhibition in vivo compared with either inhibitor alone.

[0018] Without wishing to be bound by theory, the inventors have observed that PRMT5 inhibition by a PRMT5 inhibitor, such as those described elsewhere herein, likely induces cell death in cancer tissues through DNA damage. Therefore, it was hypothesized that the therapeutic effect could be enhanced by additionally administering a therapeutic agent that promotes apoptosis, i.e., programmed cell death. In certain embodiments, a BCL-2 family inhibitor, such as ABT-199 (venetoclax), ABT-263 (navitoclax), or A-1331852, was administered in combination with a PRMT5 inhibitor. As disclosed herein, it was surprisingly found that when administered in vitro, the combination effectively inhibited tumor cell viability in a synergistic manner. As disclosed herein, it was surprisingly found that in vivo, the combination therapy suppressed tumor volume more effectively than either agent alone.

[0019] The methods provided herein can be used to treat a wide variety of cancers, including tumors (e.g., lung cancer, prostate cancer, breast cancer, brain cancer, skin cancer, cervical cancer, testicular cancer, etc.). More specifically, cancers that can be treated by the compositions and methods of the present invention include, but are not limited to, tumor types such as astrocytoma, breast cancer, cervical cancer, colorectal cancer, endometrial cancer, esophageal cancer, gastric cancer, head and neck cancer, hepatocellular carcinoma, laryngeal cancer, lung cancer, oral cancer, ovarian cancer, prostate cancer, thyroid cancer, and sarcoma. More specifically, these compounds can be used to treat the following: cardiac system: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma, and teratoma; pulmonary system: bronchogenic carcinoma (squamous cell carcinoma, undifferentiated small cell carcinoma, undifferentiated large cell carcinoma, adenocarcinoma), alveolar (bronchiolopulmonary) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondroitin hamartoma, mesothelioma; digestive system Esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, lipoma), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, smooth muscle tumors); genitourinary system: kidney (adenocarcinoma, Wilm's tumor (nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma); liver: liver cancer (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma tumor, hepatic hemangioma; biliary tract: gallbladder cancer, duodenal papilla, bile duct cancer; bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor, chordoma, osteochondroma (squamous cell carcinoma, undifferentiated small cell carcinoma, undifferentiated large cell carcinoma, adenocarcinoma), benign chondroma, chondroblastoma, chondrodysplastic fibroma, osteoid osteoma, and giant cell tumor;Nervous system: skull (osteoma, hemangioma, granuloma, xanthomas, osteitis deformans), meninges (meningioma, meningeal sarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma, embryonal tumor (pinealoma), glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), spinal cord neurofibroma, meningioma, glioma, sarcoma); gynecology: uterus (endometrial cancer), cervix (cervical cancer, preneoplastic cervical dysplasia), ovary [ovarian cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa cell tumor, Sertoli-Leydig cell tumor, dysgerminoma, malignant teratoma], vulva (Squamous cell carcinoma, carcinoma in situ, adenocarcinoma, fibrosarcoma, melanoma), granulosa cell tumor, vagina [(clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma)], fallopian tube (cancer); blood system: blood [myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative disorders, multiple myeloma, myelodysplastic syndrome], Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma); skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, dysplastic nevi, lipoma, hemangioma, dermatofibroma, keloid, psoriasis; adrenal gland: neuroblastoma;

[0020] In certain embodiments of the disclosed methods, the cancer is an MTAP-associated cancer. For example, in certain embodiments, the cancer is a cancer associated with a homozygous deletion of the MTAP gene (MTAP DEL ) The target includes, for example, MTAP DEL A subject is identified or diagnosed as having an MTAP-associated cancer when the MTAP-associated cancer gene expression level is determined using an appropriate assay or kit. Alternatively, the subject is suspected of having an MTAP-associated cancer, or the subject has clinical records indicating that the subject has an MTAP-associated cancer.

[0021] In some embodiments of any of the methods or uses described herein, an assay is used to determine the treatment eligibility of a subject using a sample (for example, a biological sample or a biopsy sample, such as a paraffin-embedded biopsy sample) from the subject.Such assays include, but are not limited to, next-generation sequencing, next-generation sequencing of circulating tumor DNA (ctDNA) in plasma, immunohistochemistry, fluorescence microscopy, FISH analysis (break-apart method), Southern blotting, Western blotting, FACS analysis and PCR-based amplification (for example, RT-PCR and quantitative real-time RT-PCR).As is well known in the art, assays are typically carried out using, for example, at least one labeled nucleic acid probe or at least one labeled antibody or its antigen-binding fragment.

[0022] In certain embodiments, the cancer in the methods of the present disclosure is selected from non-small cell lung cancer, pancreatic cancer, head and neck cancer, bladder cancer, esophageal cancer, diffuse large B-cell lymphoma, gastric cancer, melanoma, breast cancer, cholangiocarcinoma, mesothelioma, and malignant peripheral nerve sheath tumor.

[0023] In certain embodiments, the cancer in the methods of the present disclosure is selected from lung cancer (e.g., mesothelioma or non-small cell lung cancer (NSCLC) (including adenocarcinoma and squamous cell)], pancreatic cancer, colon cancer, head and neck cancer (e.g., squamous cell carcinoma (HNSCC)), bladder cancer, esophageal cancer, lymphoma (e.g., diffuse large B-cell lymphoma), gastric cancer, melanoma, breast cancer, and brain cancer (e.g., glioblastoma and glioma).

[0024] In certain embodiments, the cancer in the methods of the present disclosure is selected from lung cancer (e.g., mesothelioma, or NSCLC, including adenocarcinoma or squamous cell carcinoma), pancreatic cancer, colon cancer, head and neck cancer (e.g., squamous cell carcinoma (HNSCC)), esophageal cancer, and melanoma.

[0025] In certain embodiments, the cancer in the methods of the present disclosure is selected from mesothelioma, NSCLC (eg, adenocarcinoma and squamous cell carcinoma), pancreatic cancer, HNSCC, and colon cancer.

[0026] In one embodiment of the method of the present disclosure, cancer is lung cancer.For example, lung cancer can be NSCLC (for example, adenocarcinoma and squamous cell carcinoma) or mesothelioma.In certain embodiments, cancer is NSCLC.

[0027] In one embodiment of the disclosed methods, the cancer is pancreatic cancer.

[0028] In one embodiment of the disclosed methods, the cancer is colon cancer.

[0029] In certain embodiments described elsewhere herein, the BCL-2 family inhibitor comprises at least one of the following: the BCL-2 selective inhibitor venetoclax (ABT-199), the BCL-2 / Bcl-w / BCL-xL inhibitor navitoclax (ABT-263), A-1155463, A-1331852, obatoclax (GX15-070), ABT-737 TW-37 gossypol and (R)-(-)-gossypol, HA14-1, sabutoclax, DT2216, or a combination thereof. For example, in certain embodiments, the BCL-2 family inhibitor is navitoclax. In other embodiments, the BCL-2 family inhibitor is venetoclax. In further embodiments, the BCL-2 family inhibitor is A-1155463. In yet further embodiments, the BCL-2 family inhibitor is A-1331852. In a further embodiment, the BCL-2 family inhibitor is obatoclax. In a further embodiment, the BCL-2 family inhibitor is ABT-737. In a further embodiment, the BCL-2 family inhibitor is gossypol or (R)-(-)-gossypol. In a further embodiment, the BCL-2 family inhibitor is HA14-1. In another embodiment, the BCL-2 family inhibitor is subotoclax. In another embodiment, the BCL-2 family inhibitor is DT2216.

[0030] As provided above, venetoclax (ABT-199) (CAS Registry Number: 1257044-40-8), navitoclax (ABT-263) (CAS Registry Number: 923564-51-6), A-1155463 (CAS Registry Number: 1235034-55-5), A-1331852 (CAS Registry Number: 1430844-80-6), obatoclax (GX15-070) (CAS Registry Number: 803712-67-6), ABT-737 (CAS Registry Number: 8 52808-04-9), TW-27 (CAS Registry Number: 877877-35-5), gossypol (CAS Registry Number: 303-45-7), (R)-(-)-gossypol acetic acid (CAS Registry Number: 866541-93-7), HA14-1 (CAS Registry Number: 65673-63-4), DT2216 (CAS Registry Number: 2365172-42-3), and / or sabutoclax (CAS Registry Number: 1228108-65-3) are administered in the methods of the present disclosure. For example, venetoclax is a drug approved for oral administration. Navitoclax is a drug not approved for oral administration. A-1155463 has been administered by intraperitoneal injection (IP) in in vivo studies. A-1331852 is known to be bioavailable when administered orally. Obatoclax is administered orally and is usually administered in the form of the mesylate salt (CAS Registry Number: 803712-79-0).

[0031] As mentioned above, PRMT5 inhibitors are also administered in the method of the present disclosure.As used herein, "PRMT5 inhibitor" refers to the compound of the present disclosure described herein.These compounds can negatively regulate or inhibit all or part of the enzymatic activity of PRMT5, particularly in the presence of MTA combined in vitro or in vivo, or in cells containing high levels of MTA.In certain embodiments, PRMT5 inhibitors are MTA-cooperative PRMT5 inhibitors.

[0032] In certain embodiments, the PRMT5 inhibitor of the present disclosure is any of the PRMT5 inhibitors disclosed in International Patent Application No. PCT / US20 / 50457, filed September 11, 2020 (published on March 18, 2021 as WO 2021 / 050915 A1), which application and publication are incorporated herein by reference in their entireties. In certain other embodiments, the PRMT5 inhibitor of the present disclosure is any of the PRMT5 inhibitors disclosed in International Patent Application No. PCT / US22 / 020056, filed March 11, 2022 (published on September 15, 2022 as WO2022192745A1), which application and publication are incorporated herein by reference in their entireties.

[0033] In certain other embodiments, the PRMT5 inhibitor of the present disclosure is International Patent Application No. PCT / US22 / 035508, filed June 29, 2022 (published January 5, 2023 as WO2023278564), which application and publication are incorporated herein by reference in their entireties.

[0034] In certain other embodiments, the PRMT5 inhibitor of the present disclosure is any one of the PRMT5 inhibitors disclosed in U.S. Provisional Application No. 63 / 276,479, filed November 5, 2021, and U.S. Provisional Application No. 63 / 356,861, filed June 29, 2022, which are incorporated by reference herein in their entireties.

[0035] For example, the PRMT5 inhibitor in the methods of the disclosure described herein may be represented by Formula IIA, IIB, or IIC: [ka] [In the formula, A is CR 9 or N; D is (C(R 9 )2) 1-2 -NH2, [ka] or D is [ka] where if methylene is attached to E, then E is C; E, C, CR 9 or N; each L is independently a bond or C1-C3 alkylene; W is CR 9 or N; Each X is independently a bond, O, S, or -NR 4 -or-NR 4 C(O)-; each Z is independently a bond, —SO—, —SO2—, —CH(OH)—, or —C(O)—; Each R 2 are independently hydroxy, halogen, cyano, cyanomethyl, -(NR 4 )2, hydroxyalkyl, alkoxy, -SO2C1-C3 alkyl, X-(C1-C3 alkyl)-aryl, heteroalkyl, C2-C4 alkynyl, -X-haloalkyl, -X-C1-C5 alkyl, -Z-C1-C5 alkyl, heterocyclyl, -XL-cycloalkyl, -Z-cycloalkyl, -X-aryl, -Z-aryl or -X-heteroaryl, wherein said heterocyclyl, said cycloalkyl, said aryl and said heteroaryl are selected from the group consisting of one or more R 5 optionally substituted with; Each R 4 are independently hydrogen or C1-C3 alkyl; Each R 5are independently cyano, oxo, halogen, C1-C3 alkyl, hydroxyalkyl, hydroxy, alkoxy, alkoxy-C1-C3 alkyl, -X-haloalkyl, -Z-cycloalkyl, X-(C1-C3 alkyl)-aryl, X-(C1-C3 alkyl)-aryl substituted with cyano, -XL-cycloalkyl optionally substituted with C1-C3 alkyl or oxo, -XL-heteroaryl optionally substituted with one or more C1-C3 alkyl or oxo, -XL-heterocyclyl optionally substituted with one or more C1-C3 alkyl or oxo, or -X-aryl; R 6 is hydrogen, halogen, C1-C3 alkyl, haloalkyl, hydroxy, alkoxy, C1-C3 alkyl-alkoxy, N(R 9 )2, NR 9 C(O)R 9 , C(O)R 9 , oxetane and THF; R 7 is H or C1-C3 alkyl optionally substituted with one or more halogens; R 8 is H or C1-C3 alkyl; and Each R 9 are independently H or C1-C3 alkyl, halogen or haloalkyl. or a pharmaceutically acceptable salt thereof (embodiment 1).

[0036] Embodiment 2 relates to a PRMT5 inhibitor in the methods of the disclosure, which is represented by formula IIA: [ka] The compound is provided as follows:

[0037] Embodiment 3 relates to a PRMT5 inhibitor in the methods of the disclosure, which is represented by formula IIB: [ka] The compound is provided as follows:

[0038] Embodiment 4 relates to a PRMT5 inhibitor in the methods of the disclosure, which is represented by formula IIC: [ka] The compound is provided as follows:

[0039] In the fifth embodiment, W is CR 9 The method according to any one of embodiments 1 to 4 is provided, wherein:

[0040] In embodiment 6, A is CR 9 The method according to any one of embodiments 1 to 4 is provided, wherein:

[0041] Embodiment 7 provides the method of any one of embodiments 1 to 4, wherein E is N.

[0042] In the eighth embodiment, W is CR 9 and A is CR 9 and E is N.

[0043] Embodiment 9 is R 2 is selected from benzothiophene, naphthalene, quinoline, chroman, isochroman, dihydrobenzodioxin, indolazine, tetrahydroisoindolazine, dihydroisobenzofuran, benzene, isoquinolinone, benzodioxone, thienopyridine, tetrahydroindolone, indolizine, dihydroindolizinone, imadazopyridinone, thienopyrimidine, thiophene, pyrrolopyrimidinone, thiazolopyridinone, dihydropyrrolidine, isoindalone, and tetrahydroisoquinoline.

[0044] Embodiment 10 is a method for preparing a compound having a structure in which each R 5are independently cyano, oxo, halogen, C1-C3 alkyl, hydroxy, hydroxyalkyl, alkoxy-C1-C3 alkyl, -XL-heterocyclyl optionally substituted with one or more C1-C3 alkyl or oxo, or -XL-cycloalkyl optionally substituted with C1-C3 alkyl or oxo.

[0045] Embodiment 11 is R 6 is selected from hydrogen, hydroxy, chlorine, —NHC(O)CH3, —C(O)CF2H, —NH2, —CF2, —CH3, —O—CH2CH3, —CH2—CH2—O—CH3, oxetane, and THF.

[0046] Embodiment 12 provides the method of any one of embodiments 1-11, wherein one of L, X, and Z is a bond.

[0047] Embodiment 13 provides the method of embodiment 12, wherein L, X, and Z are all bonds.

[0048] One embodiment of the present disclosure is a compound represented by formula (IIIC): [ka] [In the formula, A is CR 9 or N; D is -CH2-NH2, [ka] and; W is CR 9 or N, where R 9 is H or C1-C3 alkyl; G, Q, J and U are C(H), C(R 5) and N, with the proviso that only one or two of G, Q, J, and U may be N; Each R 5 are independently hydroxy, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C6 cycloalkoxy, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl; R 6 is hydrogen, halogen, C1-C6 alkyl, C1-C6 haloalkyl, hydroxy, C1-C6 alkoxy, C1-C3 alkoxyC1-C3 alkyl, C3-C6 heterocycloalkyl, -C(O)-C1-C3 haloalkyl, -N(R 9 )2 or -NR 15 (CO)R 16 where each R 9 are independently H or C1-C3 alkyl, and R 15 is hydrogen or methyl, and R 16 is C1-C3 alkyl; and R 7 is C1-C3 alkyl or C1-C3 haloalkyl] (Embodiment 14) or a pharmaceutically acceptable salt thereof.

[0049] Embodiment 15 provides the method of embodiment 14, wherein A is CH.

[0050] Embodiment 16 provides the method of embodiment 14 or 15, wherein W is N.

[0051] Embodiment 17 provides the method of embodiment 14 or 15, wherein W is CH.

[0052] Embodiment 18 provides the method of any of embodiments 14-17, wherein D is -CH2-NH2.

[0053] Embodiment 19 is a case in which the PRMT5 inhibitor has the following formula: [ka] The method of the present disclosure provides a compound of embodiment 14.

[0054] Embodiment 20 is R 6 is hydrogen, halogen, C1-C6 alkyl, C1-C6 haloalkyl, hydroxy, C1-C6 alkoxy, C1-C3 alkoxyC1-C3 alkyl, C3-C6 heterocycloalkyl, -C(O)-C1-C3 haloalkyl, -N(R 9 )2 or -NR 15 (CO)R 16 The method according to any one of embodiments 14 to 19 is provided, wherein

[0055] Embodiment 21 is R 6 is hydrogen, halogen, C1-C3 alkyl, C1-C3 haloalkyl, hydroxy, C1-C3 alkoxy, C1-C3 alkoxyC1-C3 alkyl, C3-C6 heterocycloalkyl, -C(O)-C1-C3 haloalkyl, -N(R 9 )2 or -NR 15 (CO)R 16 The method according to any one of embodiments 14 to 19 is provided, wherein

[0056] Embodiment 22 is R 6 is hydrogen, chloro, fluoro, methyl, ethyl, difluoromethyl, hydroxy, methoxy, ethoxy, (methoxy)methyl, (ethoxy)methyl, (methoxy)ethyl, (ethoxy)ethyl, oxetanyl, tetrahydrofuranyl, —C(O)-difluoromethyl, —NH, or —NH(CO)CH.

[0057] Embodiment 23 is R 6 is halogen, C1-C6 alkyl, C1-C6 haloalkyl, hydroxy, C1-C6 alkoxy, C1-C3 alkoxyC1-C3 alkyl, C3-C6 heterocycloalkyl, -C(O)-C1-C3 haloalkyl, -N(R 9)2 or -NR 15 (CO)R 16 The method according to any one of embodiments 14 to 19 is provided, wherein

[0058] Embodiment 24 is R 6 is halogen, C1-C3 alkyl, C1-C3 haloalkyl, hydroxy, C1-C3 alkoxy, C1-C3 alkoxyC1-C3 alkyl, C3-C6 heterocycloalkyl, -C(O)-C1-C3 haloalkyl, -N(R 9 )2 or -NR 15 (CO)R 16 The method according to any one of embodiments 14 to 19 is provided, wherein

[0059] Embodiment 25 is R 6 is chloro, fluoro, methyl, ethyl, difluoromethyl, hydroxy, methoxy, ethoxy, (methoxy)methyl, (ethoxy)methyl, (methoxy)ethyl, (ethoxy)ethyl, oxetanyl, tetrahydrofuranyl, —C(O)-difluoromethyl, —NH, or —NH(CO)CH.

[0060] Embodiment 26 provides the method of any of embodiments 23-25, wherein each G, Q, J, and U is independently C(H).

[0061] Embodiment 27 is an embodiment in which G, Q, J and U are C(H) and C(R 5 26. The method of any of embodiments 23-25, wherein the hydroxyl group is independently selected from the group consisting of:

[0062] Embodiment 28 provides the method of any of embodiments 23-25, wherein G, Q, J, and U are independently selected from C(H) and N.

[0063] Embodiment 29 is R 6 is hydrogen; and at least one of G, Q, J and U is C(R 5 ) and the remaining G, Q, J and U are C(H), C(R5 ) and N, where each R 5 is independently hydroxy, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C6 cycloalkoxy, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl.

[0064] Embodiment 30 provides the method of embodiment 29, wherein one or two of G, Q, J and U are N.

[0065] Embodiment 31 is R 6 is hydrogen; and at least one of G, Q, J and U is C(R 5 ), and the remaining G, Q, J, and U are independently C(H) and C(R 5 ), where each R 5 is independently hydroxy, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C6 cycloalkoxy, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl.

[0066] Embodiment 32 is an embodiment in which at least one of G, Q, J, and U is C(R 5 ), and the remaining G, Q, J, and U are independently C(H); for example, only one of G, Q, J, and U is C(R 5 32. The method of claim 31, wherein

[0067] Embodiment 33 is an embodiment in which two of G, Q, J and U are C(R 5 ) and the remaining G, Q, J and U are independently C(H).

[0068] Embodiment 34 is an embodiment in which three of G, Q, J and U are C(R5 ) and the remaining G, Q, J and U are C(H).

[0069] Embodiment 35 is an embodiment in which G, Q, J and U, together with the thiophene to which they are attached, are: [ka] 20. The method of any one of embodiments 14 to 19, wherein

[0070] Embodiment 36 provides the method of embodiment 35, wherein G, Q, J, and U, together with the thiophene ring to which they are attached, form a benzo[b]thiophene.

[0071] Embodiment 37 is R 5 If there is R 5 is hydroxy, halogen, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C3-C6 cycloalkoxy, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl.

[0072] Embodiment 38 is R 5 If there is R 5 is hydroxy, halogen, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl.

[0073] Embodiment 39 is R 5 If there is R 5is hydroxy, chloro, fluoro, methyl, ethyl, methoxy, ethoxy, 2,2-difluoroethoxy, oxetanyl, tetrahydrofuranyl, (methoxy)methyl, (ethoxy)methyl, (methoxy)ethyl, or (ethoxy)ethyl.

[0074] Embodiment 40 is R 7 The method of any one of embodiments 14-39, wherein is methyl.

[0075] Embodiment 41 is R 7 The method of any one of embodiments 14-39, wherein is ethyl.

[0076] Embodiment 42 is R 7 The method of any one of embodiments 14-39, wherein is propyl (eg, isopropyl).

[0077] Embodiment 43 is R 7 Embodiment 40 provides the method of any one of embodiments 14-39, wherein is difluoromethyl or trifluoromethyl.

[0078] Embodiment 44 is a method for treating a PRMT5 inhibitor comprising administering to a patient having the following formula: [ka] [In the formula, G, Q, J and U together with the thiophene attached thereto form: [ka] (In the formula, each R 5 are independently hydroxy, halogen, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl. Forming; and R 6is hydrogen, halogen, C1-C3 alkyl, C1-C3 haloalkyl, hydroxy, C1-C3 alkoxy, C1-C3 alkoxyC1-C3 alkyl, C3-C6 heterocycloalkyl, -C(O)-C1-C3 haloalkyl, -N(R 9 )2 or -NR 15 (CO)R 16 is] 15. The method of embodiment 14, wherein

[0079] Embodiment 45 is a method for treating a PRMT5 inhibitor comprising administering to a patient having the following formula: [ka] [In the formula, G, Q, J and U together with the thiophene bonded thereto form a group of the following formula: [ka] (In the formula, each R 5 are independently hydroxy, halogen, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl. Forming: and R 6 is halogen, C1-C3 alkyl, C1-C3 haloalkyl, hydroxy, C1-C3 alkoxy, C1-C3 alkoxyC1-C3 alkyl, C3-C6 heterocycloalkyl, -C(O)-C1-C3 haloalkyl, -N(R 9 )2 or -NR 15 (CO)R 16 is] 15. The method of embodiment 14, wherein

[0080] Embodiment 46 is a method for treating a PRMT5 inhibitor comprising administering to a patient having the following formula: [ka] [In the formula, G, Q, J and U together with the thiophene bonded thereto form a group of the following formula: [ka] (In the formula, each R 5 are independently hydroxy, halogen, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl. form] 15. The method of embodiment 14, wherein

[0081] Embodiment 47 is a method for treating a PRMT5 inhibitor comprising administering to a patient of formula (IIIB): [ka] [In the formula, A is CR 9 or N; D is -CH2-NH2, [ka] and; W is CR 9 or N, where R 9 is H or C1-C3 alkyl; R 51 is hydrogen, fluoro, chloro or methyl, or R 51 and R 52 together with the atoms to which they are attached form a C4-C6 heterocycloalkyl (e.g., hydrofuranyl); R 52 is fluoro, chloro or methyl, or R 52 and R 53 together with the atoms to which they are attached to form phenyl; R 53 is hydrogen, fluoro, chloro or methyl; R 54is hydrogen, halogen, C1-C3 alkyl or C1-C3 alkoxy; L 5 is -O- or -CH2-; R 6 is hydrogen, halogen, C1-C6 alkyl, hydroxy, C1-C6 alkoxy, C1-C3 alkoxyC1-C3 alkyl, C3-C6 heterocycloalkyl, -C(O)-C1-C3 haloalkyl or -NR 15 (CO)R 16 where R 15 is hydrogen or methyl, and R 16 is C1-C3 alkyl; R 7 is C1-C3 alkyl or C1-C3 haloalkyl] or a pharmaceutically acceptable salt thereof.

[0082] Embodiment 48 is A is -CH or -CCH; D is -CH2-NH2; W is -CH, -CCH3 or N; R 51 、 R 52 , R 53 and R 54 are each independently selected from hydrogen, fluoro, chloro, or methyl; L 5 is -O-; R 6 is hydrogen, fluoro, chloro or methyl; and R 7 is C1-C2 alkyl or C1-C2 haloalkyl; 48. The method of embodiment 47 is provided.

[0083] Embodiment 49 is A and W are -CH; D is -CH2-NH2; R 51 , R 52 and R 53are each independently selected from hydrogen, fluoro, chloro, and methyl; R 54 is hydrogen; L 5 is -O-; R 6 is hydrogen; and R 7 is methyl, The method of embodiment 47 or embodiment 48 is provided.

[0084] Embodiment 50 is A and W are -CH; D is -CH2-NH2; R 51 and R 52 are each independently selected from fluoro, chloro, and methyl; R 53 and R 54 is hydrogen; L 5 is -O-; R 6 is hydrogen; and R 7 is methyl, The method according to any one of embodiments 47 to 49 is provided.

[0085] Embodiment 51 provides the method of embodiment 47, wherein A is CH.

[0086] Embodiment 52 provides the method of embodiment 47 or 48, wherein W is N.

[0087] Embodiment 53 provides the method of embodiment 47 or 48, wherein W is CH.

[0088] Embodiment 54 provides the method of any of embodiments 47-50, wherein D is -CH2-NH2.

[0089] Embodiment 55 is R 54is hydrogen or methyl.

[0090] Embodiment 56 is R 54 52. The method of any one of embodiments 47-51, wherein is hydrogen.

[0091] Embodiment 57 is R 54 The method of any one of embodiments 47-51, wherein is methyl.

[0092] Embodiment 58 is a method for treating a PRMT5 inhibitor comprising administering to a patient a compound of the following formula: [ka] for example, [ka] 48. The method of embodiment 47, wherein

[0093] Embodiment 59 is L 5 The method of any one of embodiments 47-55, wherein is —CH 2 —.

[0094] Embodiment 60 is L 5 The method of any one of embodiments 47-55, wherein is —O—.

[0095] Embodiment 61 is R 6 is hydrogen, halogen, C1-C3 alkyl, C1-C3 haloalkyl, hydroxy, C1-C3 alkoxy, C1-C3 alkoxyC1-C3 alkyl, C3-C6 heterocycloalkyl, -C(O)-C1-C3 haloalkyl, -N(R 9 )2 or -NR 15 (CO)R 16 For example, R 6is hydrogen, chloro, fluoro, methyl, ethyl, difluoromethyl, hydroxy, methoxy, ethoxy, (methoxy)methyl, (ethoxy)methyl, (methoxy)ethyl, (ethoxy)ethyl, oxetanyl, tetrahydrofuranyl, —C(O)-difluoromethyl, —NH, or —NH(CO)CH.

[0096] Embodiment 62 is R 6 is hydrogen, halogen, C1-C6 alkyl or C1-C6 alkoxy; for example, R 6 is hydrogen, halogen, C1-C3 alkyl, or C1-C3 alkoxy.

[0097] Embodiment 63 is R 6 58. The method of any one of embodiments 47-57, wherein is hydrogen, chloro, fluoro, methyl, ethyl, methoxy, or ethoxy.

[0098] Embodiment 64 is R 6 is halogen, C1-C3 alkyl, C1-C3 haloalkyl, hydroxy, C1-C3 alkoxy, C1-C3 alkoxyC1-C3 alkyl, C3-C6 heterocycloalkyl, -C(O)-C1-C3 haloalkyl, -N(R 9 )2 or -NR 15 (CO)R 16 For example, R 6 is chloro, fluoro, methyl, ethyl, difluoromethyl, hydroxy, methoxy, ethoxy, (methoxy)methyl, (ethoxy)methyl, (methoxy)ethyl, (ethoxy)ethyl, oxetanyl, tetrahydrofuranyl, —C(O)-difluoromethyl, —NH, or —NH(CO)CH.

[0099] Embodiment 65 is R 6 is halogen, C1-C6 alkyl or C1-C6 alkoxy; for example, R 6is halogen, C1-C3 alkyl, or C1-C3 alkoxy.

[0100] Embodiment 66 is R 6 58. The method of any one of embodiments 47-57, wherein is chloro, fluoro, methyl, ethyl, methoxy, or ethoxy.

[0101] Embodiment 67 is R 7 The method of any one of embodiments 47-63, wherein is methyl.

[0102] Embodiment 68 is R 7 The method of any one of embodiments 47-63, wherein is ethyl.

[0103] Embodiment 69 is R 7 The method of any one of embodiments 47-63, wherein is propyl (eg, isopropyl).

[0104] Embodiment 70 is R 7 Embodiment 64. The method of any one of embodiments 47-63, wherein is difluoromethyl or trifluoromethyl.

[0105] Embodiment 71 is R 53 is hydrogen or methoxy; or R 53 68. The method according to any of embodiments 47-67, wherein is hydrogen.

[0106] Embodiment 72 is a method for treating a PRMT5 inhibitor comprising administering to a patient having the following formula: [ka] 48. The method of embodiment 47, wherein

[0107] Embodiment 73 is R 52 is fluoro and R 51Embodiment 69 provides the method of any one of embodiments 47-69, wherein is hydrogen, fluoro, chloro, or methyl.

[0108] Embodiment 74 is R 52 is fluoro and R 51 The method of any one of embodiments 47-69, wherein is chloro.

[0109] Embodiment 75 is R 52 is fluoro and R 51 is methyl or hydrogen (e.g., R 52 is fluoro and R 51 is methyl; or R 52 is fluoro and R 51 is hydrogen).

[0110] Embodiment 76 is R 51 and R 52 together with the atoms to which they are attached form a hydrofuranyl (e.g., [ka] 70. The method of any one of embodiments 47 to 69, wherein

[0111] Embodiment 77 is a method for treating the PRMT5 inhibitor comprising administering to a patient: [ka] The method of any one of embodiments 47 to 76 is provided, wherein

[0112] Embodiment 78 is a method for treating the PRMT5 inhibitor comprising administering to a patient: [ka] The method of any one of embodiments 47 to 77 is provided, wherein

[0113] One embodiment of the present disclosure is a compound represented by formula (IIIA): [ka] [In the formula, A is CR 9 or N; D is -CH2-NH2, [ka] and; W is CR 9 or N, where R 9 is H or C1-C3 alkyl; R 2 is the following: [ka] (In the formula, R 56 is hydrogen, fluoro, chloro or methyl; G, Q, J and U are C(H), C(R 5 ) and N, with the proviso that only one or two of G, Q, J, and U may be N; Each R 5 are independently hydroxy, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C6 cycloalkoxy, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl. and; R 6 is hydrogen, halogen, C1-C6 alkyl, hydroxy, C1-C6 alkoxy, C1-C3 alkoxyC1-C3 alkyl, C3-C6 heterocycloalkyl, -C(O)-C1-C3 haloalkyl or -NR 15 (CO)R 16 where R 15 is hydrogen or methyl, and R 16 is C1-C3 alkyl; and R 7is C1-C3 alkyl or C1-C3 haloalkyl] (embodiment 79) or a pharmaceutically acceptable salt thereof.

[0114] One embodiment of the present disclosure is a compound represented by formula (IIIA): [ka] [In the formula, A is CR 9 or N; D is -CH2-NH2, [ka] and; W is CR 9 or N, where R 9 is H or C1-C3 alkyl; R 2 teeth, [ka] (In the formula, R 56 is hydrogen, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C1-C6 haloalkoxy) and; R 6 is hydrogen, halogen, C1-C6 alkyl, hydroxy, C1-C6 alkoxy, C1-C3 alkoxyC1-C3 alkyl, C3-C6 heterocycloalkyl, -C(O)-C1-C3 haloalkyl or -NR 15 (CO)R 16 where R 15 is hydrogen or methyl, and R 16 is C1-C3 alkyl; and R 7 is C1-C3 alkyl or C1-C3 haloalkyl] (embodiment 80) or a pharmaceutically acceptable salt thereof.

[0115] Embodiment 81 provides the method of embodiment 79 or 80, wherein A is CH.

[0116] Embodiment 82 provides the method of embodiment 79 or 80, wherein W is N.

[0117] Embodiment 83 provides the method of embodiment 79 or 80, wherein W is CH.

[0118] Embodiment 84 provides the method of either embodiment 79 or 80, wherein D is —CH 2 —NH 2 .

[0119] Embodiment 85 is directed to a compound having the following formula: [ka] 81. The method of embodiment 79 or 80, wherein

[0120] Embodiment 86 is R 2 but, [ka] The method of any one of embodiments 79 and 81 to 85 is provided, wherein

[0121] Embodiment 87 is an embodiment in which G, Q, J and U are C(H) and C(R 5 87. The method of embodiment 86, wherein the hydroxyl group is independently selected from:

[0122] Embodiment 88 provides the method of embodiment 86, wherein G, Q, J and U are independently C(H).

[0123] Embodiment 89 is an embodiment in which at least one of G, Q, J, and U is C(R 5 ) and the remaining G, Q, J and U are independently C(H); for example, only one of G, Q, J and U is C(R 5 87. The method of embodiment 86, wherein

[0124] Embodiment 90 is an embodiment in which U is N and G, Q, and J are C(H) and C(R 5 87. The method of embodiment 86, wherein the hydroxyl group is independently selected from:

[0125] Embodiment 91 is an embodiment in which G is N and Q, J, and U are C(H) and C(R 5 87. The method of embodiment 86, wherein the hydroxyl group is independently selected from:

[0126] Embodiment 92 is R 5 If there is R 5 is hydroxy, halogen, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C3-C6 cycloalkoxy, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl.

[0127] Embodiment 93 is R 5 If there is R 5 is hydroxy, halogen, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl.

[0128] Embodiment 94 is R 5 If there is R 5 is hydroxy, chloro, fluoro, methyl, ethyl, methoxy, ethoxy, 2,2-difluoroethoxy, oxetanyl, tetrahydrofuranyl, (methoxy)methyl, (ethoxy)methyl, (methoxy)ethyl, or (ethoxy)ethyl.

[0129] Embodiment 95 is R 5 If there is R 5is halogen, C1-C6 alkyl or C1-C6 alkoxy; for example, R 6 is halogen, C1-C3 alkyl, or C1-C3 alkoxy.

[0130] Embodiment 96 is R 5 If there is R 5 is chloro, fluoro, methyl, ethyl, methoxy, or ethoxy.

[0131] Embodiment 97 is R 56 The method of any one of embodiments 79 or 81-91, wherein is fluoro, chloro, or methyl.

[0132] Embodiment 98 is R 2 but, [ka] The method according to any one of embodiments 80 to 85, wherein

[0133] Embodiment 99 is R 56 is hydrogen, fluoro, chloro, or methyl.

[0134] Embodiment 100 is R 6 is hydrogen, halogen, C1-C3 alkyl, C1-C3 haloalkyl, hydroxy, C1-C3 alkoxy, C1-C3 alkoxyC1-C3 alkyl, C3-C6 heterocycloalkyl, -C(O)-C1-C3 haloalkyl, -N(R 9 )2 or -NR 15 (CO)R 16 For example, R 6is hydrogen, chloro, fluoro, methyl, ethyl, difluoromethyl, hydroxy, methoxy, ethoxy, (methoxy)methyl, (ethoxy)methyl, (methoxy)ethyl, (ethoxy)ethyl, oxetanyl, tetrahydrofuranyl, —C(O)-difluoromethyl, —NH, or —NH(CO)CH.

[0135] Embodiment 101 is R 6 is hydrogen, halogen, C1-C6 alkyl or C1-C6 alkoxy; for example, R 6 is hydrogen, halogen, C1-C3 alkyl, or C1-C3 alkoxy.

[0136] Embodiment 102 is R 6

[0033] Embodiment 99 provides the method of any one of embodiments 79-99, wherein is hydrogen, chloro, fluoro, methyl, ethyl, methoxy, or ethoxy.

[0137] Embodiment 103 is R 6 is halogen, C1-C3 alkyl, C1-C3 haloalkyl, hydroxy, C1-C3 alkoxy, C1-C3 alkoxyC1-C3 alkyl, C3-C6 heterocycloalkyl, -C(O)-C1-C3 haloalkyl, -N(R 9 )2 or -NR 15 (CO)R 16 For example, R 6 is chloro, fluoro, methyl, ethyl, difluoromethyl, hydroxy, methoxy, ethoxy, (methoxy)methyl, (ethoxy)methyl, (methoxy)ethyl, (ethoxy)ethyl, oxetanyl, tetrahydrofuranyl, —C(O)-difluoromethyl, —NH, or —NH(CO)CH.

[0138] Embodiment 104 is R 6 is halogen, C1-C6 alkyl or C1-C6 alkoxy; for example, R 6is halogen, C1-C3 alkyl, or C1-C3 alkoxy.

[0139] Embodiment 105 is R 6 99. The method of any one of embodiments 79-99, wherein is chloro, fluoro, methyl, ethyl, methoxy, or ethoxy.

[0140] Embodiment 106 is R 7 The method of any one of embodiments 79-105, wherein is methyl.

[0141] Embodiment 107 is R 7 The method of any one of embodiments 79-105, wherein is ethyl.

[0142] Embodiment 108 is R 7 The method of any one of embodiments 79-105, wherein is propyl (eg, isopropyl).

[0143] Embodiment 109 is R 7 Embodiment 106. The method of any one of embodiments 79-105, wherein is difluoromethyl or trifluoromethyl.

[0144] In certain embodiments of the methods of the present disclosure described herein, the PRMT5 inhibitor is [ka] is.

[0145] In certain embodiments of the methods of the present disclosure described herein, the PRMT5 inhibitor is [ka] is.

[0146] In certain embodiments of the methods of the present disclosure described herein, the PRMT5 inhibitor is [ka] is.

[0147] In certain embodiments of the methods of the present disclosure described herein, the PRMT5 inhibitor is [ka] is.

[0148] In one aspect, the disclosure provides a method for treating rheumatoid arthritis by administering to a patient a therapeutically effective amount of ABT-199 (venetoclax), wherein ABT-199 is [ka] is); and A therapeutically effective amount of the following formula: [ka] The present invention provides a method for treating cancer in a subject, comprising administering to the subject a PRMT5 inhibitor of the formula:

[0149] In one aspect, the present disclosure provides a method for treating atopic dermatitis comprising administering to a subject a therapeutically effective amount of ABT-263 (navitoclax), wherein ABT-263 is [ka] is); and A therapeutically effective amount of the following formula: [ka] The present invention provides a method for treating cancer in a subject, comprising administering to the subject a PRMT5 inhibitor of the formula:

[0150] The PRMT5 inhibitors and / or BCL-2 family inhibitors of the present disclosure (e.g., ABT-199 (venetoclax), ABT-263 (navitoclax), A-1155463, A-1331852, obatoclax (GX15-070), ABT-737, TW-37, gossypol or (R)-(-)-gossypol, HA14-1, sabutoclax, and / or DT2216) may be provided as a pharmaceutical composition comprising a therapeutically effective amount of the inhibitor and a pharmaceutically acceptable carrier, excipient, and / or diluent. The PRMT5 inhibitors and / or BCL-2 family inhibitors of the present disclosure may be formulated by any method known in the art and may be prepared for administration by any route (e.g., including, but not limited to, parenteral, oral, sublingual, transdermal, topical, nasal, intratracheal, or rectal).

[0151] The characteristics of the carrier vary depending on the route of administration. As used herein, the term "pharmaceutically acceptable" refers to a non-toxic material that is compatible with biological systems such as cells, cell cultures, tissues, or organisms, and does not inhibit the effectiveness of the biological activity of the active ingredient. Thus, the pharmaceutical composition of the present disclosure may contain, in addition to the inhibitor, diluents, fillers, salts, buffers, stabilizers, solubilizers, and other substances well known in the art. The preparation of pharmaceutically acceptable formulations can be carried out, for example, by the methods described in Remington's Pharmaceutical Sciences, 18 th Edition, ed. A. Gennaro, Mack Publishing Co., Easton, Pa., 1990.

[0152] The PRMT5 inhibitors and BCL-2 family inhibitors of the present disclosure are administered in therapeutically effective amounts. As used herein, the phrase "therapeutically effective amount" or "effective amount" refers to the amount of an active agent that induces the biological or pharmaceutical response desired by a researcher, physician or other clinician in a tissue, system, subject or human. Generally, a therapeutically effective amount is an amount sufficient to provide a biological or pharmacological response to a subject without causing serious toxic effects. The dose of the active agent is determined based on the recipient's body weight (m 2 ) about 1 to 500 mg / m per day 2 , e.g., 5 to 400 mg / m per day 2 , more commonly 10–300 mg / m per day 2 Typical topical dosages range from 0.01 to 10% wt / wt in a suitable carrier.

[0153] In certain embodiments of the disclosed methods, the therapeutically effective amount of the PRMT5 inhibitor ranges from about 0.01 to 300 mg / kg per day. For example, in certain embodiments, the therapeutically effective amount of the PRMT5 inhibitor ranges from about 0.1 to 100 mg / kg per day, or 25 to 100 mg / kg per day, or 50 to 100 mg / kg per day.

[0154] In certain embodiments, the therapeutically effective amount of a PRMT5 inhibitor is less than 1%, e.g., less than 10%, less than 25%, or less than 50% of the clinically established therapeutic amount (e.g., the amount required when the PRMT5 inhibitor is administered alone).

[0155] In certain embodiments of the disclosed methods, the therapeutically effective amount of a BCL-2 family inhibitor is administered in a dose that is in accordance with the recipient's body weight (m 2 ) about 1 to 500 mg / m per day 2 , e.g., 5-400 mg / m per day 2 , more commonly 10–300 mg / m per day 2For example, in certain embodiments, a therapeutically effective amount of a BCL-2 family inhibitor ranges from about 30 to 300 mg / m per day. 2 (e.g., 50–250 mg / m per day) 2 , 50-200 mg / m 2 or 50-150 mg / m 2 ) range.

[0156] For example, in various embodiments, the BCL-2 family inhibitor can be ABT-199 (venetoclax), ABT-263 (navitoclax), A-1155463, A-1331852, obatoclax (GX15-070), ABT-737, TW-37, gossypol or (R)-(-)-gossypol, HA14-1, sabutoclax, or DT2216. Accordingly, in certain embodiments of the methods of the present disclosure, a therapeutically effective amount of each of these agents is administered based on the recipient's body weight (m 2 ) about 1 to 500 mg / m per day 2 , e.g., 5-400 mg / m per day 2 , more commonly 10–300 mg / m per day 2 For example, in certain embodiments, a therapeutically effective amount of any one of these agents ranges from about 30 to 300 mg / m per day. 2 (e.g., 50–250 mg / m per day) 2 , 50-200 mg / m 2 or 50-150 mg / m 2 ) range.

[0157] In certain embodiments, the therapeutically effective amount of ABT-199 (venetoclax), ABT-263 (navitoclax), A-1155463, A-1331852, obatoclax (GX15-070), ABT-737, TW-37, gossypol or (R)-(-)-gossypol, HA14-1, sabutoclax, or DT2216 is less than 1% (e.g., less than 10%, less than 25%, less than 50%, or less than 75%) of the clinically established therapeutic amount (e.g., the amount required when the compound is administered alone).

[0158] In combination therapy, the PRMT5 inhibitor and BCL-2 family inhibitor of the present disclosure [e.g., ABT-199 (venetoclax), ABT-263 (navitoclax), A-1155463, A-1331852, obatoclax (GX15-070), ABT-737, TW-37, gossypol or (R)-(-)-gossypol, HA14-1, sabutoclax, or DT2216] are intended to encompass sequential administration of each agent in a regimen that provides the beneficial effects of the drug combination (e.g., the PRMT5 inhibitor and BCL-2 family inhibitor of the present disclosure can be formulated as separate compositions that are administered in sequence), and are also intended to encompass co-administration of these agents in a substantially simultaneous manner, such as in a single dosage form comprising a fixed ratio of these active agents, or in multiple or individual dosage forms of each agent. The present disclosure is not limited in the order of administration: a PRMT5 inhibitor of the present disclosure may be administered before or after (i.e., sequentially) the administration of a BCL-2 family inhibitor of the present disclosure, or at the same time (i.e., simultaneously) as the administration of a BCL-2 family inhibitor of the present disclosure.

[0159] The methods of the present disclosure are useful as first-line treatments. Thus, in certain embodiments of the methods of the present disclosure, the subject has not previously received another first-line treatment.

[0160] The method of the present disclosure is also useful as a first-line maintenance treatment or a second-line treatment. Thus, in certain embodiments of the method of the present disclosure, the subject has previously completed another first-line treatment. For example, in certain embodiments, the method of the present disclosure can provide a delay in cancer progression and recurrence in a subject who has previously completed another first-line chemotherapy. For example, in certain embodiments, the subject has previously completed a treatment regimen, including but not limited to chemotherapy, targeted therapy, or immunotherapy, as a monotherapy or in combination with other therapies. In certain embodiments of the method of the present disclosure, the subject has previously completed another first-line chemotherapy and has a partial response to such chemotherapy.

[0161] (definition) For simplicity, chemical moieties are defined and referred to throughout primarily as monovalent chemical moieties (e.g., alkyl, aryl, etc.). However, such terms can be used to convey corresponding multivalent moieties under appropriate structural circumstances apparent to those skilled in the art. For example, an "alkyl" moiety generally refers to a monovalent group (e.g., CH3-CH2-), but under certain circumstances a divalent linking moiety may be "alkyl," in which case one skilled in the art would understand the alkyl to be the divalent group equivalent to the term "alkylene" (e.g., -CH2-CH2-). Similarly, in situations where a divalent moiety is required and "aryl" is described, one skilled in the art would understand the term "aryl" to refer to the corresponding divalent moiety, arylene. All atoms are understood to have the normal valences required for bond formation (i.e., carbon is tetravalent, nitrogen is trivalent, oxygen is divalent, and S is divalent, tetravalent, or hexavalent depending on oxidation state).

[0162] The term "amino" refers to -NH2.

[0163] The term "acetyl" refers to "-C(O)CH3".

[0164] As used herein, the term "acyl" refers to an alkylcarbonyl or arylcarbonyl substituent, wherein said alkyl and said aryl moieties are as defined herein.

[0165] As used herein, the term "alkyl" refers to saturated straight- and branched-chain aliphatic groups having 1 to 12 carbon atoms. Thus, "alkyl" includes C1, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24, C25, C26, C27, C28, C29, C30, C31, C32, C33, C34, C35, C36, C37, C38, C39, C40, C41, C42, C43, C44, C45, C46, ​​C47, C48, C49, C50, C51, C52, C53, C54, C55, C56, 10 , C 11 and C 12 Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl.

[0166] As used herein, the term "alkenyl" refers to an unsaturated, straight- or branched-chain aliphatic group having one or more carbon-carbon double bonds and having 2 to 12 carbon atoms. Thus, "alkenyl" includes C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24, C25, C26, C27, C28, C29, C30, C31, C32, C33, C34, C35, C36, C37, C38, C39, C40, C41, C42, C43, C44, C45, C46, ​​C47, C48, C49, C50, C51, C52, 10 , C 11 and C 12 Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, and hexenyl.

[0167] As used herein, the term "alkynyl" refers to an unsaturated, straight- or branched-chain aliphatic group having 2 to 12 carbon atoms and one or more carbon-carbon triple bonds. Thus, "alkynyl" refers to any of C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24, C25, C26, C27, C28, C29, C30, C31, C32, C33, C34, C35, C36, C37, C38, C39, C40, C41, C42, C43, C44, C45, C46, ​​C47, C48, C49, C50, C51, C5 10 , C 11 and C 12 Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, and hexynyl.

[0168] An "alkylene," "alkenylene," or "alkynylene" group is an alkyl, alkenyl, or alkynyl group, as defined herein, that is positioned between and functions to link two other chemical groups. Examples of alkylene groups include, but are not limited to, methylene, ethylene, propylene, and butylene. Exemplary alkenylene groups include, but are not limited to, ethenylene, propenylene, and butenylene. Exemplary alkynylene groups include, but are not limited to, ethynylene, propynylene, and butynylene.

[0169] The term "alkoxy" refers to -OC1-C6 alkyl.

[0170] As used herein, the term "cycloalkyl" refers to saturated and partially unsaturated cyclic hydrocarbon groups having 3 to 12 carbon atoms. Thus, "cycloalkyl" refers to C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24, C25, C26, C27, C28, C29, C30, C31, C32, C33, C34, C35, C36, C37, C38, C39, C40, C41, C42, C43, C44, C45, C46, ​​C47, C48, C49, C50, C51, C52, C53, C54, C55, C56, C57, C 10 , C 11 and C 12 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.

[0171] The term "heteroalkyl" refers to an alkyl group, as defined hereinabove, wherein one or more of the carbon atoms in the chain is O, S, or NR x are independently substituted with R x is hydrogen or C1-C3 alkyl. Examples of heteroalkyl groups include methoxymethyl, methoxyethyl, and methoxypropyl.

[0172] An "aryl" group is a C6-C aryl group containing one to three aromatic rings. 14 The aromatic moiety. Thus, "aryl" includes C6, C 10 , C 13 and C 14 Cyclic hydrocarbon groups are included. Exemplary aryl groups include C-C 10 An aryl group. Particular aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, and fluorenyl. "Aryl" groups also include fused polycyclic (e.g., bicyclic) ring systems in which one or more of the fused rings are non-aromatic, provided that at least one ring is aromatic (e.g., indenyl).

[0173] An "aralkyl" or "arylalkyl" group comprises an aryl group covalently bonded to an alkyl group, which is in turn bonded to another group through the alkyl portion. Exemplary aralkyl groups include -(C-C)alkyl (C-C 10) aryl, including, but not limited to, benzyl, phenethyl, and naphthylmethyl. For example, arC1-C3 alkyl is an aryl group covalently linked to a C1-C3 alkyl.

[0174] A "heterocyclyl" or "heterocycle" group is a monocyclic or bicyclic (fused or spiro) ring structure having 3 to 12 atoms (3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 atoms) (e.g., 4 to 8 atoms), where one or more ring atoms are independently -C(O)-, N, NR 4 , O, or S, and the remainder of the ring atoms are quaternary carbon or carbonyl carbons. Examples of heterocyclic groups include, but are not limited to, epoxy, oxiranyl, oxetanyl, azetidinyl, aziridinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, piperazinyl, imidazolidinyl, thiazolidinyl, thiatanyl, dithianyl, trithianyl, azathianyl, oxathianyl, dioxolanyl, oxazolidinyl, oxazolidinonyl, decahydroquinolinyl, piperidonyl, 4-piperidonyl, thiomorpholinyl, dimethylmorpholinyl, and morpholinyl. Specifically excluded from the scope of this term are compounds having adjacent ring O and / or S atoms.

[0175] As used herein, "L-heterocyclyl" refers to a heterocyclyl group covalently linked to another group via an alkylene linker.

[0176] As used herein, the term "heteroaryl" refers to a group having 5 to 14 ring atoms, preferably 5, 6, 10, 13, or 14 ring atoms; having 6, 10, or 14 pi electrons shared in a cyclic arrangement; and having, in addition to carbon atoms, 1 to 3 heteroatoms, each independently N, O, or S. "Heteroaryl" also includes fused polycyclic (e.g., bicyclic) ring systems in which one or more of the fused rings is non-aromatic, provided that at least one ring is aromatic and at least one ring contains an N, O, or S ring atom.Examples of heteroaryl groups include acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzo[d]oxazol-2(3H)-one, 2H-benzo[b][1,4]oxazin-3(4H)-one, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, furanyl, furanyl, and the like. Zanyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazo Allyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathinyl, phenoxazinyl, phthalazinyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclinyl thiazinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl and xanthenyl.

[0177] An "L-heteroaralkyl" or "L-heteroarylalkyl" group comprises a heteroaryl group covalently linked to another group via an alkylene linker. Examples of heteroalkyl groups include C1-C6 alkyl groups and heteroaryl groups having 5, 6, 9, or 10 ring atoms. Examples of heteroaralkyl groups include pyridylmethyl, pyridylethyl, pyrrolylmethyl, pyrrolylethyl, imidazolylmethyl, imidazolylethyl, thiazolylmethyl, thiazolylethyl, benzimidazolylmethyl, benzimidazolylethylquinazolinylmethyl, quinolinylmethyl, quinolinylethyl, benzofuranylmethyl, indolinylethylisoquinolinylmethyl, isoindolylmethyl, cinnolinylmethyl, and benzothiophenylethyl. Specifically excluded from the scope of this term are compounds having adjacent ring O and / or S atoms.

[0178] An "arylene," "heteroarylene," or "heterocyclylene" group is a divalent aryl, heteroaryl, or heterocyclyl group, respectively, as defined herein, that is positioned between and serves to link two other chemical groups.

[0179] As used herein, when a moiety (e.g., cycloalkyl, aryl, heteroaryl, heterocyclyl, urea, etc.) is described as "optionally substituted" without specifying the substituents, it means that the group optionally has 1 to 4, preferably 1 to 3, and more preferably 1 or 2 non-hydrogen substituents.

[0180] As used herein, the term "halogen" or "halo" refers to chlorine, bromine, fluorine or iodine.

[0181] The term "haloalkyl" refers to an alkyl chain in which one or more hydrogens have been replaced by halogen. Exemplary haloalkyls are trifluoromethyl, difluoromethyl, fluorochloromethyl, chloromethyl, and fluoromethyl.

[0182] The term "hydroxyalkyl" refers to -alkylene-OH. [Example]

[0183] The methods of the present disclosure are further illustrated by the following examples, which are not to be construed as limiting the scope or spirit of the disclosure to the particular procedures and compounds described therein.

[0184] Example 1 - In vivo combination study (LU99 model) Immunodeficient female nu / nu mice were given 5x10 6 LU99 cancer cells / 50% Matrigel were implanted. Tumor volume was measured using a caliper and calculated using the formula: 0.5 × L × W 2 (where L is the length of the tumor; W is the width of each tumor). 3 Upon reaching 100 mg / kg / day, animals were randomized to receive A) vehicle, B) PRMT5 inhibitor, C) navitoclax (ABT-263), D) venetoclax (ABT-199), E) A-1331852, F) PRMT5 inhibitor and navitoclax, F) PRMT5 inhibitor and venetoclax, or G) PRMT5 inhibitor and A-1331852, all administered orally (PO) at the indicated doses and schedules for 20–34 days. Tumor volumes were measured twice weekly (n=5 / treatment group). The mean tumor volume and standard error of the mean for each study day were calculated and plotted using GraphPad Prism.

[0185] The PRMT5 inhibitor was MRTX1719, administered at 100 mg / kg once daily (QD). MRTX1719 is (2M)-2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile, and is disclosed as Example 16-8 on page 307 of International Patent Publication No. WO 2021 / 050915 A1, published March 18, 2021, and incorporated herein by reference in its entirety.

[0186] The BCL-2 family inhibitors ABT-199 (venetoclax) and ABT-263 (navitoclax) used in this example were supplied by Selleck Chemicals (Houston, TX) and were administered once daily (QD) at 100 mg / kg.

[0187] The results are shown in FIG. [Table 1]

[0188] Combining MRTX1719 with each of ABT-199 (venetoclax) and ABT-263 (navitoclax) resulted in increased antitumor activity compared to either compound alone, as measured by the change in tumor volume over time in a xenograft model derived from the LU99 cell line.

[0189] Example 2 - In vivo combination study (NCI-H1437 model) Example 2 was carried out according to the in vivo testing procedure described in Example 1 above, using xenografts derived from the NCI-H1437 cell line.

[0190] The PRMT5 inhibitor was MRTX1719, administered at 100 mg / kg once daily (QD). MRTX1719 is (2M)-2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile, and is disclosed as Example 16-8 on page 307 of International Patent Publication No. WO 2021 / 050915 A1, published March 18, 2021, and incorporated herein by reference in its entirety. ABT-263 (navitoclax), a BCL-2 family inhibitor, was supplied by Selleck Chemicals (Houston, TX), and administered at 100 mg / kg once daily (QD). A-1331852, a BCL-2 family inhibitor, was synthesized at Wuxi AppTec (Wuhan, China) and administered once daily (QD) at 25 mg / kg.

[0191] The results are shown in FIG. [Table 2]

[0192] MRTX1719, in combination with ABT-263 (navitoclax) and A-1331852, exhibited enhanced antitumor activity as measured by changes in tumor volume over time in a xenograft model derived from the NCI-H1437 cell line.

[0193] Example 3 - In vivo combination study (SW1573 model) Example 3 was carried out using xenografts derived from the SW1573 cell line according to the in vivo testing procedure described in Example 1 above.

[0194] The PRMT5 inhibitor was MRTX1719, administered at 100 mg / kg once daily (QD). MRTX1719 is (2M)-2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile, and is disclosed as Example 16-8 on page 307 of International Patent Publication No. WO 2021 / 050915 A1, published March 18, 2021, and incorporated herein by reference in its entirety. ABT-263 (navitoclax), a BCL-2 family inhibitor, was supplied by Selleck Chemicals (Houston, TX), and administered at 100 mg / kg once daily (QD). A-1331852, a BCL-2 family inhibitor, was synthesized at Wuxi AppTec (Wuhan, China) and administered once daily (QD) at 25 mg / kg.

[0195] The results are shown in FIG. [Table 3]

[0196] Combining MRTX1719 with ABT-263 (navitoclax) and A-1331852 resulted in increased antitumor activity compared with either compound alone, as measured by changes in tumor volume over time in a xenograft model derived from the SW173 cell line.

[0197] Example 4 - In vivo combination study (LU99 model) Example 4 was carried out according to the in vivo testing procedures described in Example 1 above.

[0198] The PRMT5 inhibitor was MRTX1719, administered at 100 mg / kg once daily (QD). MRTX1719 is (2M)-2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile, and is disclosed as Example 16-8 on page 307 of International Patent Publication No. WO 2021 / 050915 A1, published March 18, 2021, and incorporated herein by reference in its entirety. A-1331852, a BCL-2 family inhibitor, was synthesized by Wuxi AppTec (Wuhan, China) and administered at 25 mg / kg twice daily (BID).

[0199] The results are shown in FIG. [Table 4]

[0200] Combining MRTX1719 with A-1331852 resulted in increased antitumor activity compared with either compound alone, as judged by the change in tumor volume over time in a xenograft model derived from the LU99 cell line.

[0201] Example 5 - In vivo combination study (NCI-H1650 model) Example 5 was carried out using xenografts derived from the NCI-H1650 cell line according to the in vivo testing procedure described in Example 1 above.

[0202] The PRMT5 inhibitor was MRTX1719, administered at 100 mg / kg once daily (QD). MRTX1719 is (2M)-2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile, and is disclosed as Example 16-8 on page 307 of International Patent Publication No. WO 2021 / 050915 A1, published March 18, 2021, and incorporated herein by reference in its entirety. ABT-263 (navitoclax), a BCL-2 family inhibitor, was supplied by Selleck Chemicals (Houston, TX), and administered at 100 mg / kg once daily (QD). A-1331852, a BCL-2 family inhibitor, was synthesized at Wuxi AppTec (Wuhan, China) and administered at 25 mg / kg twice daily (BID).

[0203] The results are shown in FIG. [Table 5]

[0204] Combining MRTX1719 with ABT-263 (navitoclax) and A-1331852 resulted in increased antitumor activity compared with either compound alone, as measured by tumor volume change over time in a xenograft model derived from the NCI-H1650 cell line.

[0205] Example 6 - Synergy scores, in vitro combination studies This example demonstrates that exemplary PRMT5 inhibitor compounds of the type described in WO2021 / 050915, when used in combination with a BCL-2 family inhibitor, synergistically inhibit the growth of MTAP-deficient cancer cell lines.

[0206] To investigate whether combining a BCL-2 family inhibitor with an exemplary PRMT5 inhibitor disclosed herein would result in synergistic activity, a panel of MTAP-deficient cancer cell lines was generated.

[0207] Assays to determine pairwise combination synergy scores for each cell line were performed in triplicate. Cells of a specific MTAP-deficient cell line were seeded in the appropriate growth medium for that cell line (e.g., RPMI 1640 medium supplemented with 10% FBS and cell line-specific reagents required for growth) into additional wells of a 384- or 96-well plate and a separate 384- or 96-well control plate to determine baseline luminescence. Plates were incubated overnight at 37°C in a 5% CO2 atmosphere.

[0208] Cell-Titer Glo reagent (CTG; Promega Corporation) was added to each designated baseline well, and the plate was incubated with shaking at room temperature for 20 minutes. Baseline luminescence was quantified using a BMG ClarioStar multimode plate reader according to the manufacturer's instructions.

[0209] 1000X drug serial dilutions in 100% DMSO were prepared, including 9 single-agent 3-fold dilutions of an exemplary PRMT5 inhibitor (of the type described in WO2021 / 050915) at a top dose of 3000 nM and 6 single-agent 5-fold dilutions of a BCL-2 family inhibitor at a top dose of 3000 nM as reference standards.

[0210] A 10X intermediate dose plate containing single-agent dilutions of exemplary PRMT5 inhibitors (of the type described in WO2021 / 050915) and / or BCL-2 family inhibitors was prepared in serum-free medium. Additionally, a 54-dilution matrix combining exemplary PRMT5 inhibitors (of the type described in WO2021 / 050915) and BCL-2 family inhibitors was prepared as test samples.

[0211] 10X single agent and 54 combined dose matrices were added to corresponding wells of a 384-well plate seeded with the appropriate cell line, and the plate was incubated for 120 hours at 37°C in a 5% CO2 atmosphere. Cell-Titer Glo reagent (CTG) was added to each test well, and the plate was incubated with shaking at room temperature for 20 minutes, after which luminescence was quantified using a BMG ClarioStar multimode plate reader according to the manufacturer's instructions. To normalize test samples, a baseline luminescence determined from each test sample prior to analysis was subtracted, and average values ​​were calculated from multiple samples.

[0212] The raw data and generated metadata were used as input files to calculate the % effect for each treatment condition and analyzed using four independent mathematical reference models designed to determine whether two test compounds exhibit synergistic effects: Loewe additivity, Bliss independence, Highest Single Agent and ZIP (reference).

[0213] The data output from each mathematical model is the assignment of a relative synergy score. The data reported in Table 6 is the sum of the Loewe additivity, Bliss independence, Highest Single Agent and ZIP synergy scores ("Composite Synergy Score"). [Table 6]

[0214] These results demonstrate that the combination of various BCL-2 family inhibitors and exemplary PRMT5 inhibitor compounds (of the type described in WO2021 / 050915) exhibited synergistic effects in each cell line, thereby enhancing the sensitivity of MTAP-deficient cell lines to PRMT5 inhibitors.

[0215] It is understood that the examples and embodiments described herein are for illustrative purposes and that various modifications or variations therein will be suggested to those skilled in the art and are to be incorporated within the spirit and scope of this application and the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference for all purposes.

Claims

1. A combination drug for treating cancer in a target, comprising a therapeutically effective amount of a BCL-2 family inhibitor and a therapeutically effective amount of a protein arginine N-methyltransferase 5 (PRMT5) inhibitor.

2. The combination pharmaceutical according to claim 1, wherein the cancer is associated with homozygous deletion of the methylthioadenosine phosphorylase (MTAP) gene.

3. The combination pharmaceutical according to claim 1 or 2, wherein the PRMT5 inhibitor is a methylthioadenosine (MTA) cooperative PRMT5 inhibitor.

4. The combination pharmaceutical product according to claim 1 or 2, wherein the cancer is lung cancer, non-small cell lung cancer, pancreatic cancer, colon cancer, bladder cancer, head and neck cancer, esophageal cancer, diffuse large B-cell lymphoma, lymphoma, gastric cancer, melanoma, breast cancer, brain tumor, bile duct cancer, mesothelioma, or malignant peripheral nerve sheath tumor.

5. The combination drug according to claim 4, wherein the cancer is mesothelioma.

6. The combination drug according to claim 4, wherein the cancer is non-small cell lung cancer.

7. PRMT5 inhibitors are expressed as follows: 【Chemistry 1】 The combination pharmaceutical according to claim 1 or 2.

8. PRMT5 inhibitors are expressed as follows: 【Chemistry 2】 The combination pharmaceutical according to claim 1 or 2, wherein the compound represented by or a pharmaceutically acceptable salt thereof.

9. PRMT5 inhibitors are expressed as follows: 【Transformation 3】 The combination pharmaceutical according to claim 1 or 2, wherein the compound represented by or a pharmaceutically acceptable salt thereof.

10. PRMT5 inhibitors are expressed as follows: 【Chemistry 4】 The combination pharmaceutical according to claim 1 or 2, wherein the compound represented by or a pharmaceutically acceptable salt thereof.

11. The combination pharmaceutical according to claim 1 or 2, wherein the therapeutically effective amount of PRMT5 inhibitor is in the range of approximately 0.01 to 300 mg / kg per day.

12. The combination pharmaceutical according to claim 1 or 2, wherein the therapeutically effective amount of PRMT5 inhibitor is in the range of approximately 0.1 to 100 mg / kg per day.

13. The combination pharmaceutical according to claim 1 or 2, wherein the therapeutically effective amount of PRMT5 inhibitor is less than 1% of a clinically established therapeutic dose, for example, less than 10%, less than 25%, or less than 50%.

14. The combination pharmaceutical according to claim 1 or 2, wherein the BCL-2 family inhibitor is selected from one or more of the following: ABT-199 (venetoclax), ABT-263 (navitoclax), A-1155463, A-1331852, ovatoclax (GX15-070), ABT-737, TW-37, gossypol, (R)-(-)-gossypol, HA14-1, subtoclax, and DT2216.

15. The combination pharmaceutical according to claim 1 or 2, wherein the BCL-2 family inhibitor comprises ABT-199.

16. The combination pharmaceutical according to claim 15, wherein the BCL-2 family inhibitor is ABT-199.

17. The combination pharmaceutical according to claim 1 or 2, wherein the BCL-2 family inhibitor comprises ABT-263.

18. The combination drug according to claim 17, wherein the BCL-2 family inhibitor is ABT-263.

19. A therapeutically effective dose of a BCL-2 family inhibitor is approximately 1 to 500 mg / m² per day. ２ A combination pharmaceutical according to claim 1 or 2, which is within the range of [the specified range].

20. A therapeutically effective dose of a BCL-2 family inhibitor is approximately 10-300 mg / m² per day. ２ A combination pharmaceutical according to claim 1 or 2, which is within the range of [the specified range].

21. The combination pharmaceutical according to claim 1 or 2, wherein the therapeutically effective amount of BCL-2 family inhibitor is less than 1% of a clinically established therapeutic dose, for example, less than 10%, less than 25%, or less than 50%.

22. The combination pharmaceutical according to claim 1 or 2, wherein a BCL-2 family inhibitor and a PRMT5 inhibitor are included in a single dosage form.

23. The combination pharmaceutical according to claim 1 or 2, wherein the BCL-2 family inhibitor and the PRMT5 inhibitor are included in separate compositions.

24. The combination pharmaceutical according to claim 1 or 2, wherein a BCL-2 family inhibitor and a PRMT5 inhibitor are administered sequentially.

25. The combination pharmaceutical according to claim 1 or 2, wherein a BCL-2 family inhibitor and a PRMT5 inhibitor are administered simultaneously.

26. The combination drug according to claim 1 or 2, wherein the subject has previously received or completed first-line chemotherapy.

27. The combination drug according to claim 26, wherein the first-line chemotherapy is gemcitabine, nab-paclitaxel, 5-FU, irinotecan, oxaliplatin, capecitabine, cisplatin, carboplatin, fludarabine, cyclophosphamide, fluoropyrimidine, pemetrexed, doxorubicin, ifosfamide, epirubicin, or paclitaxel.

28. A therapeutically effective dose of ABT-199 (venetoclax): 【Transformation 5】 ; and The following formula indicates a therapeutically effective amount: 【Transformation 6】 A combination drug for treating cancer in the target population, comprising a PRMT5 inhibitor as indicated by [the formula].

29. A therapeutically effective dose of ABT-263 (Navitoclax): 【Transformation 7】 ; and The following formula indicates a therapeutically effective amount: 【Transformation 8】 A combination drug for treating cancer in the target population, comprising a PRMT5 inhibitor as indicated by [the formula].

30. The combination pharmaceutical according to claim 28, wherein ABT-199 (venetoclax) and a PRMT5 inhibitor are included in a single dosage form.

31. The combination pharmaceutical according to claim 28, wherein ABT-199 (venetoclax) and a PRMT5 inhibitor are included in separate compositions.

32. The combination pharmaceutical according to claim 29, wherein ABT-263 (navitoclax) and a PRMT5 inhibitor are included in a single dosage form.

33. The combination pharmaceutical according to claim 29, wherein ABT-263 (Navitoclax) and a PRMT5 inhibitor are included in separate compositions.

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