Combination therapy for treating cancer using PRMT5 inhibitors and BCL-2

Abstract

Disclosed herein are methods of treating cancer. More specifically, the disclosure provides methods of treating cancer in an individual using compounds that are PRMT5 inhibitors, particularly in combination with BCL-2 family inhibitors.

Description

Background of the Invention

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to U.S. Provisional Application No. 63 / 403,201 filed on September 1, 2022 and U.S. Provisional Application No. 63 / 497,022 filed on April 19, 2023, the disclosures of each of which are incorporated herein by reference in their entirety. Field of the Invention

[0004] The present disclosure relates to methods of treating cancer. The present disclosure also relates to methods of treating cancer in an individual with 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.

[0005] Related technologies

[0006] PRMT5 is a type II arginine methyltransferase that catalyzes the transfer of a methyl group from S-adenosyl-L-methionine (SAM) to the ω-nitrogen of the guanidino functional group of L-arginine residues in proteins (ω-monomethylation) and the transfer of a second methyl group to another ω-nitrogen, generating symmetric dimethylarginine (sDMA). PRMT5 forms a complex with methylosome protein 50 (MEP50), which is required for substrate recognition and orientation, and is also required for PRMT5-dependent SDMA modification of histone 2A and histone 4 (e.g., see Ho et al. (2013) PLoS ONE 8(2):e57008).

[0007] Homozygous deletions of p16 / CDKN2a are common in cancer, and these mutations often involve co-deletion of adjacent genes, including the gene encoding methylthioadenosine phosphorylase (MTAP). It is estimated that about 15% of all human cancers have homozygous deletions of the MTAP gene (e.g., see Firestone & Schramm (2017) J. Am. Chem Soc. 139 (39): 13754-13760).

[0008] Cells lacking MTAP activity have elevated levels of MTAP substrate, methylthioadenosine (MTA), which is a potent inhibitor of PRMT5. Inhibition of PRMT5 activity by elevated MTA results in reduced methylation activity and increased sensitivity of cell proliferation to further PRMT5 depletion or activity inhibition. Thus, loss of MTAP activity reduces the methylation activity of PRMT5, rendering cells selectively dependent on PRMT5 activity.

[0009] Despite the importance of PRMT5 for cell viability and its ubiquity in cancer, effective therapies that inhibit PRMT5 have been elusive. Therefore, there remains a need to develop new PRMT5 inhibitor therapies to treat a wide range of cancers.

[0010] BCL-2 family inhibitors are being studied for the treatment of cancer. BCL-2 family inhibitors are speculated to work by inhibiting pro-survival signaling mediated by selected BCL-2 family members. It is expected that the inhibition of BCL-2 family members will block the survival of cancer cells and provide clinical benefits to cancer patients. The Bcl-2 inhibitor venetoclax is approved for the treatment of chronic lymphocytic leukemia. The Bcl-2, Bcl-xL and Bcl-w inhibitors navitoc are currently undergoing clinical trials for many liquid and solid cancers. Navitoc is currently being studied as a single active agent and combined with therapies including chemotherapy. The Bcl-xL inhibitor DT2216 is being studied for the treatment of various solid and liquid cancers. It is expected that BCL-2 family inhibitors have different utilities for the treatment of various cancers, depending on the sensitivity of various cancer types and those cancers to the inhibition of one or more BCL-2 family members.

[0011] Despite the importance of BCL-2 family members for cancer cell survival and their prevalence in cancer therapy resistance, effective therapies that inhibit BCL-2 family members have been difficult to find. Therefore, there remains a need to improve the efficacy of cancer treatments involving the administration of BCL-2 family inhibitors. SUMMARY OF THE INVENTION

[0013] The present disclosure provides a method for treating cancer in an individual. The method includes administering a therapeutically effective amount of a PRMT5 inhibitor to an individual and a therapeutically effective amount of a BCL-2 family inhibitor. The BCL-2 family inhibitor in such a combination and / or combination therapy can be selected from one or more of the following: ABT-199 (Venetoclax), ABT-263 (Navitoclax), A-1155463, A-1331852, Obakla (GX15-070), ABT-737, TW-37, cotton pol or (R)-(-)-cotton pol, HA14-1, sabutoclax, DT2216, AMG176, PRT1419, AZD5991, S64315 / MIK665 or a combination of these. Additional BCL-2 family inhibitors are currently being developed and can be used in the embodiments of the present invention described herein. The PRMT5 inhibitor can be, for example, any PRMT5 inhibitor disclosed in WO 2021 / 050915 A1, including MRTX-1719.

[0014] Also provided herein are methods for treating cancer in an individual identified as being in need thereof. The methods include the step of determining whether the individual's cancer is associated with a homozygous deletion of MTAP (eg, an MTAP-associated cancer).

[0015] These and other features and advantages of the present invention will be more fully understood from the following detailed description and the appended claims. Note that the scope of the claims is defined by the description therein and not by the specific discussion of the features and advantages set forth in this specification. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] The accompanying drawings provide a further understanding of the method of the present disclosure, and are incorporated into and constitute a part of this specification. The accompanying drawings illustrate one or more embodiments of the present disclosure, and together with the description serve to explain the principles and operations of the present disclosure.

[0018] Figure 1 The results of Example 1 are illustrated, wherein MRTX1719 (100 mg / kg PO, QD), BCL-2 selective inhibitor venetoclax (ABT-199) (100 mg / kg PO, QD) or a combination thereof, or wherein MRTX1719 (100 mg / kg PO, QD), BCL-2 / BCL-xL inhibitor navitoc (ABT-263) (100 mg / kg PO, QD) or a combination thereof, were administered to mice with LU99 xenograft tumors (n=5 / group). Data are shown as mean tumor volume + / - SEM.

[0019] Figure 2 The results of Example 2 are illustrated, wherein MRTX1719 (100 mg / kg PO, QD), BCL-2 / Bcl-w / BCL-xL inhibitor navitoclax (ABT-263) (100 mg / kg PO, QD) or a combination thereof, or wherein MRTX1719 (100 mg / kg PO, QD), BCL-xL selective inhibitor A-1331852 (25 mg / kg PO, QD) or a combination thereof, was administered to mice with NCI-H1437 xenograft tumors (n=4 / group). Data are shown as mean tumor volume + / - SEM.

[0020] Figure 3The results of Example 3 are illustrated, wherein MRTX1719 (100 mg / kg PO, QD), BCL-2 / Bcl-w / BCL-xL inhibitor navitoclax (ABT-263) (100 mg / kg PO, QD), or a combination thereof, or wherein MRTX1719 (100 mg / kg PO, QD), BCL-xL selective inhibitor A-1331852 (25 mg / kg PO, QD), or a combination thereof, were administered to mice with SW1573 xenograft tumors (n=4 / group). Data are shown as mean tumor volume + / - SEM.

[0021] Figure 4 The results of Example 4 are illustrated, in which MRTX1719 (100 mg / kg PO, QD), BCL-xL selective inhibitor A-1331852 (25 mg / kg PO, BID), or a combination thereof, were administered to mice bearing LU99 xenograft tumors (n=5 / group). Data are shown as mean tumor volume + / - SEM.

[0022] Detailed description of the disclosure

[0023] Before describing the disclosed methods and materials, it should be understood that the aspects described herein are not limited to specific embodiments and therefore can certainly vary. It should also be understood that the terminology used herein is for the purpose of describing specific aspects only and is not intended to be limiting unless specifically defined herein.

[0024] In view of the present disclosure, the methods and compositions described herein can be configured by one of ordinary skill in the art to meet the desired needs. The present disclosure provides improvements in cancer treatment in individuals. As used herein, the terms "individual" or "patient" are used interchangeably and refer to any animal, including mammals, and most preferably humans.

[0025] The PRMT5 inhibitors disclosed herein demonstrate selective activity in MTAP-deficient cancers by binding to and further inhibiting PRMT5 when bound to the intracellular metabolite MTA. As described above, MTAP is an enzyme in the methionine salvage pathway, and its absence in cancer cells leads to the accumulation of MTA in these cells. PRMT5 is an essential enzyme required for cell survival, and therefore, the PRMT5 inhibitors disclosed herein represent a new approach to selectively treat MTAP-deficient cancers.

[0026] Single mutation may not cause cancer - most commonly, the cause of cancer formation is multiple mutations. The inventors found that treating certain cancers with PRMT5 inhibitors is improved by using combination therapy. In particular, the inventors were surprised to find that the combination therapy of MTA-cooperative PRMT5 inhibitors and BCL-2 family inhibitors (e.g., ABT-199 (venetoclax), ABT-263 (navitoclax), A-1155463, A-1331852, Obakla (GX15-070), ABT-737, TW-37, cotton pol or (R)-(-)-cotton pol, HA14-1, sabutoclax and / or DT2216) inhibits in vitro tumor cell viability in a synergistic manner, and provides stronger tumor growth inhibition in vivo compared to any single inhibitor.

[0027] Without wishing to be bound by theory, the inventors have observed that PRMT5 inhibition, such as by a PRMT5 inhibitor described in addition herein, may induce cell death in cancerous tissues through DNA damage. Therefore, it is assumed that additional therapeutic agents that enhance apoptosis or programmed cell death may be used to enhance the therapeutic effect. In certain embodiments, for example, BCL-2 family inhibitors such as ABT-199 (Venetoclax), ABT-263 (Navitoclax) or A-1331852 are administered in combination with a PRMT5 inhibitor. As disclosed herein, it is surprising to find that the in vitro combination effectively inhibits tumor cell viability in a synergistic manner. As disclosed herein, it is surprising to find that the in vivo combination effectively inhibits tumor volume to a greater extent than any single active agent alone.

[0028] The methods provided herein can be used to treat a variety of cancers, including tumors, such as 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 astrocytic carcinoma, 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, and thyroid cancer and sarcomas. More specifically, these compounds can be used to treat: Heart: sarcomas (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyosarcoma, fibroma, lipoma, and teratoma; Lung: bronchial carcinoma (squamous cell carcinoma, undifferentiated small cell carcinoma, undifferentiated large cell carcinoma, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, enchondromatous hamartoma, mesothelioma; Gastrointestinal: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, pancreatic Intestinal (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), Colon (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma); Genitourinary tract: Kidney (adenocarcinoma, Wilms' tumor (Nephroblastoma), lymphoma, leukemia), Bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), Prostate (adenocarcinoma, sarcoma), Testis (seminoma, teratoma, embryonal carcinoma, teratoma Cancer, choriocarcinoma, sarcoma, mesenchymal cell carcinoma, fibroma, fibroadenoma, adenomatoid tumor, lipoma); Liver: hepatoma (hepatocellular carcinoma), bile duct cancer, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Biliary tract: gallbladder cancer, ampullary cancer, bile duct cancer; Bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticular cell sarcoma), multiple myeloma, malignant giant cell tumor chordoma, osteochondroma (osteocartilaginous exostosis), benign enchondroma, chondroblastoma, Chondromyxofibroma, osteoid osteoma, and giant cell tumor; Nervous system: skull (osteomas, hemangiomas, granulomas, xanthomas, osteitis deformans), meninges (meningiomas, meningosarcomas, gliomas), brain (astrocytomas, medulloblastomas, gliomas, ependymomas, germ cell tumors (pinealomas), glioblastomas multiforme, oligodendrogliomas, schwannomas, retinoblastomas, congenital tumors), spinal neurofibromas, meningiomas, gliomas, sarcomas);Gynecology: Uterus (endometrial cancer), cervix (cervical cancer, preneoplastic cervical dysplasia), ovary (ovarian cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa-thecal cell tumors, Sertoli-Leydig cell tumors Tubal: Hematology: Blood (Myeloid Leukemia (Acute and Chronic), Acute Lymphocytic 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, Moles, Dysplastic Nevi, Lipoma, Hemangioma, Dermatofibroma, Keloid, Psoriasis; and Adrenal: Neuroblastoma.

[0029] In certain embodiments of the methods of the present disclosure, the cancer is an MTAP-associated cancer. For example, in certain embodiments, the cancer comprises a homozygous deletion of the MTAP gene (MTAP DEL ). An individual can be identified or diagnosed as having a MTAP-associated cancer, where, for example, MTAP is determined using a suitable assay or kit. DEL Alternatively, the individual is suspected of having a MTAP-associated cancer or the individual has a clinical record indicating that the individual has a MTAP-associated cancer.

[0030] In some embodiments of any method or use described herein, an assay is used to determine individual treatment eligibility using a sample from an individual (e.g., a biological sample or a biopsy sample, such as a paraffin-embedded biopsy sample). The assay includes, but is not limited to, next generation sequencing, next generation sequencing of circulating tumor DNA (ctDNA) in plasma, immunohistochemistry, fluorescence microscopy, fragmentation FISH analysis, Southern blotting, western blotting, FACS analysis, and PCR-based amplification (e.g., RT-PCR and quantitative real-time RT-PCR). As is well known in the art, the assay is typically performed, for example, with at least one labeled nucleic acid probe or at least one labeled antibody or antigen-binding fragment thereof.

[0031] 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, bile duct cancer, mesothelioma, and malignant peripheral nerve sheath tumor.

[0032] 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 multiforme and glioma).

[0033] In certain embodiments, the cancer in the methods of the present disclosure is selected from lung cancer (e.g., mesothelioma or NSCLC, including adenocarcinoma and squamous cell), pancreatic cancer, colon cancer, head and neck cancer (e.g., squamous cell carcinoma (HNSCC)), esophageal cancer, and melanoma.

[0034] In certain embodiments, the cancer in the methods of the present disclosure is selected from mesothelioma, NSCLC (eg, adenocarcinoma and squamous cell), pancreatic cancer, HNSCC, and colon cancer.

[0035] In one embodiment of the methods of the present disclosure, the cancer is lung cancer. For example, the lung cancer can be NSCLC (e.g., adenocarcinoma and squamous cell) or mesothelioma. In certain embodiments, the cancer is NSCLC.

[0036] In one embodiment of the disclosed methods, the cancer is pancreatic cancer.

[0037] In one embodiment of the disclosed methods, the cancer is colon cancer.

[0038] In certain embodiments as further described herein, the BCL-2 family inhibitor includes at least one of the following: BCL-2 selective inhibitor venetoclax (ABT-199), BCL-2 / Bcl-w / BCL-xL inhibitor navitoclax (ABT-263), A-1155463, A-1331852, obaclax (GX15-070), ABT-737TW-37 cottonpol and (R)-(-)-cottonpol, HA14-1, sabutoclax, DT2216, or a combination thereof. For example, in a specific embodiment, 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 still further embodiments, the BCL-2 family inhibitor is A-1331852. In further embodiments, the BCL-2 family inhibitor is obaclax. In another embodiment, the BCL-2 family inhibitor is ABT-737. In another embodiment, the BCL-2 family inhibitor is gossypol or (R)-(-)-gossypol. In another embodiment, the BCL-2 family inhibitor is HA14-1. In another embodiment, the BCL-2 family inhibitor is sabutoclax. In another embodiment, the BCL-2 family inhibitor is DT2216.

[0039] As described above, in the methods of the present disclosure, venetoclax (ABT-199) (CAS Registry Number: 1257044-40-8), navitoc (ABT-263) (CAS Registry Number: 923564-51-6), A-1155463 (CAS Registry Number: 1235034-55-5), A-1331852 (CAS Registry Number: 1430844-80-6), obacla (GX15-070) (CAS Registry Number: 803712-67-6), ABT-737 (CAS Registry Number: 852808-04-9), TW-27 (CAS Registry Number: 877877-35-5), cottony phenol (CAS Registry Number: 303-45-7 ), ((R)-(-)-gossypol acetic acid (CAS Reg. No.: 866541-93-7), HA14-1 (CAS Reg. No.: 65673-63-4), DT2216 (CAS Reg. No.: 2365172-42-3) and / or sabutoclax (CAS Reg. No.: 1228108-65-3). For example, venetoclax is an approved drug for oral administration. Navetoclax is an unapproved drug 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. Obaklax is typically administered orally in the form of its mesylate salt (CAS Reg. No.: 803712-79-0).

[0040] As described above, PRMT5 inhibitors are also administered in the methods of the present disclosure. "PRMT5 inhibitors" as used herein refer to compounds of the present disclosure as described herein. These compounds can negatively regulate or inhibit the enzymatic activity of all or part of PRMT5, particularly in vitro or in vivo in the presence of bound MTA, or in cells containing elevated levels of MTA. In certain embodiments, the PRMT5 inhibitor is an MTA-cooperative PRMT5 inhibitor.

[0041] In certain embodiments, the PRMT5 inhibitor of the present disclosure is any of the PRMT5 inhibitors disclosed in International Patent Application PCT / US20 / 50457 filed on September 11, 2020 (published as WO 2021 / 050915A1 on March 18, 2021), which are incorporated herein by reference in their entirety. In certain other embodiments, the PRMT5 inhibitor of the present disclosure is any of the PRMT5 inhibitors disclosed in International Patent Application PCT / US22 / 020056 filed on March 11, 2022 (published as WO2022192745A1 on September 15, 2022), which are incorporated herein by reference in their entirety.

[0042] In certain other embodiments, the PRMT5 inhibitor of the present disclosure is any one of the PRMT5 inhibitors disclosed in international patent application PCT / US22 / 035508 filed on June 29, 2022 (published as WO2023278564 on January 5, 2023), which application and publication are incorporated herein by reference in their entirety.

[0043] In certain other embodiments, the PRMT5 inhibitor of the present disclosure is any of the PRMT5 inhibitors disclosed in U.S. Provisional Application Nos. 63 / 276,479, filed on November 5, 2021, and 63 / 356,861, filed on June 29, 2022, which are incorporated herein by reference in their entirety.

[0044] For example, the PRMT5 inhibitor in the methods of the present disclosure as described herein is a compound of Formula IIA, IIB or IIC (Embodiment 1):

[0045]

[0046] or a pharmaceutically acceptable salt thereof, wherein:

[0047] A is for CR 9 or N;

[0048] D is (C (R 9 ) 2 ) 1-2 -NH 2 , Or D is wherein the methylene group is bonded to E, wherein E is C;

[0049] E is for C, CR 9 or N;

[0050] L is each independently a bond or C 1 -C 3 Alkylene;

[0051] W is CR 9 or N;

[0052] X is independently a bond, O, S, -NR 4 -or-NR 4 C(O)-;

[0053] Z is each independently a bond, -SO-, -SO 2 -, -CH(OH)- or -C(O)-;

[0054] R 2 are independently hydroxy, halogen, cyano, cyanomethyl, -(NR 4 ) 2, hydroxyalkyl, alkoxy, -SO 2 C 1 -C 3 Alkyl, X-(C 1 -C 3 alkyl)-aryl, heteroalkyl, C 2 -C 4 Alkynyl, -X-haloalkyl, -XC 1 -C 5 Alkyl, -ZC 1 -C 5 alkyl, heterocyclyl, -XL-cycloalkyl, -Z-cycloalkyl, -X-aryl, -Z-aryl or -X-heteroaryl, wherein the heterocyclyl, cycloalkyl, aryl and heteroaryl are optionally substituted by one or more R 5 replace;

[0055] R 4 are independently hydrogen or C 1 -C 3 alkyl;

[0056] R 5 are independently cyano, oxo, halogen, C 1 -C 3 Alkyl, hydroxyalkyl, hydroxy, alkoxy, alkoxy-C 1 -C 3 alkyl, -X-haloalkyl, -Z-cycloalkyl, X-(C 1 -C 3 Alkyl)-aryl, X-(C 1 -C 3 alkyl)-aryl, optionally C 1 -C 3 -XL-cycloalkyl substituted with alkyl or oxo, optionally with one or more C 1 -C 3 -XL-heteroaryl substituted with alkyl or oxo, optionally with one or more C 1 -C 3 -XL-heterocyclyl substituted with alkyl or oxo, or -X-aryl;

[0057] R 6 is hydrogen, halogen, C 1 -C 3 Alkyl, haloalkyl, hydroxy, alkoxy, C 1 -C 3 Alkyl-alkoxy, N(R 9 ) 2 NR 9 C(O)R 9 、C(O)R 9 , oxetane and THF;

[0058] R 7 is H or C optionally substituted by one or more halogens 1 -C 3 alkyl;

[0059] R 8 Is H or C 1 -C 3 alkyl; and

[0060] R 9 Each independently is H or C 1 -C 3 Alkyl, halogen or haloalkyl.

[0061] Embodiment 2 provides the PRMT5 inhibitor in the method of the present disclosure, which is a compound of formula IIA:

[0062]

[0063] Embodiment 3 provides the PRMT5 inhibitor in the method of the present disclosure, which is a compound of formula IIB:

[0064]

[0065] Embodiment 4 provides the PRMT5 inhibitor in the method of the present disclosure, which is a compound of formula IIC:

[0066]

[0067] Embodiment 5 provides the method of any one of embodiments 1-4, wherein W is CR 9 .

[0068] Embodiment 6 provides the method of any one of embodiments 1-4, wherein A is CR 9 .

[0069] Embodiment 7 provides the method of any one of embodiments 1-4, wherein E is N.

[0070] Embodiment 8 provides the method of any one of embodiments 1-7, wherein W is CR 9 , A is CR 9 , and E is N.

[0071] Embodiment 9 provides the method of any one of embodiments 1-8, wherein R 2Selected from the group consisting of benzothiophene, naphthalene, quinoline, chroman, isochroman, dihydrobenzodioxine, indolazine, tetrahydroindolazine, dihydroisobenzofuran, benzene, isoquinolinone, benzodioxone, thienopyridine, tetrahydroindolinone, indolizine, dihydroindolizinone, imidazopyridone, thienopyrimidine, thiophene, pyrrolopyrimidone, thiazolopyridone, dihydropyrrolizine, isoindalone and tetrahydroisoquinoline.

[0072] Embodiment 10 provides the method of any one of embodiments 1-8, wherein R 5 are independently cyano, oxo, halogen, C1-C 3 alkyl, hydroxy, hydroxyalkyl, alkoxy-C1-C3 alkyl, -XL-heterocyclyl optionally substituted with one or more C1-C3 alkyl or oxo, 3 alkyl or oxo-substituted-XL-cycloalkyl.

[0073] Embodiment 11 provides the method of any one of embodiments 1-8, wherein R 6 Selected from hydrogen, hydroxyl, chlorine, -NHC(O)CH 3 、-C(O)CF 2 H, -NH 2 , -CF 2 、-CH 3 、-O-CH 2 CH 3 、-CH 2 -CH 2 -O-CH 3 , oxetane and THF.

[0074] Embodiment 12 provides the method of any one of embodiments 1-11, wherein one of L, X and Z is a bond.

[0075] Embodiment 13 provides the method of embodiment 12, wherein L, X and Z are all bonds.

[0076] One aspect of the present disclosure provides a method wherein the PRMT5 inhibitor is a compound of formula (IIIC) (Embodiment 14):

[0077]

[0078] or a pharmaceutically acceptable salt thereof, wherein

[0079] A is for CR 9 or N;

[0080] D is -CH 2 -NH 2 ,

[0081] W is CR 9 or N, where R 9 Is H or C 1 -C 3 alkyl;

[0082] G, Q, J and U are independently selected from C(H), C(R 5 ) and N, provided that only one or two of G, Q, J and U may be N;

[0083] R 5 are independently hydroxy, halogen, C 1 -C 6 Alkyl, C 1 -C 6 Haloalkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Halogenated alkoxy, C 3 -C 6 Cycloalkoxy, C 3 -C 6 Cycloalkyl, C 3 -C 6 Heterocycloalkyl or C 1 -C 3 Alkoxy C 1 -C 3 alkyl;

[0084] R 6 is hydrogen, halogen, C 1 -C 6 Alkyl, C 1 -C 6 Haloalkyl, hydroxyl, C 1 -C 6 Alkoxy, C 1 -C 3 Alkoxy C 1 -C 3 Alkyl, C 3 -C 6 Heterocycloalkyl, -C(O)-C 1 -C 3 Haloalkyl, -N(R 9 ) 2 or -NR 15 (CO)R 16 ,

[0085] Where R 9Each independently is H or C 1 -C 3 Alkyl, R 15 is hydrogen or methyl, and R 16 It is C 1 -C 3 alkyl; and

[0086] R 7 It is C 1 -C 3 Alkyl or C 1 -C 3 Halogenated alkyl.

[0087] Embodiment 15 provides a method according to embodiment 14, wherein A is CH.

[0088] Embodiment 16 provides a method according to embodiment 14 or 15, wherein W is N.

[0089] Embodiment 17 provides a method according to embodiment 14 or 15, wherein W is CH.

[0090] Embodiment 18 provides a method according to any one of embodiments 14-17, wherein D is -CH 2 -NH 2 .

[0091] Embodiment 19 provides the methods of the present disclosure, wherein the PRMT5 inhibitor is a compound according to the following formula of Embodiment 14:

[0092]

[0093] Embodiment 20 provides a method according to any one of embodiments 14-19, wherein R 6 It is hydrogen, halogen, C 1 -C 6 Alkyl, C 1 -C 6 Haloalkyl, hydroxyl, C 1 -C 6 Alkoxy, C 1 -C 3 Alkoxy C 1 -C 3 Alkyl, C 3 -C 6 Heterocycloalkyl, -C(O)-C 1 -C 3 Haloalkyl, -N(R 9 ) 2 or -NR 15 (CO)R 16 .

[0094] Embodiment 21 provides a method according to any one of embodiments 14-19, wherein R 6 It is hydrogen, halogen, C 1 -C 3 Alkyl, C 1 -C 3 Haloalkyl, hydroxyl, C 1 -C 3 Alkoxy, C 1 -C 3 Alkoxy C 1 -C 3 Alkyl, C 3 -C 6 Heterocycloalkyl, -C(O)-C 1 -C 3 Haloalkyl, -N(R 9 ) 2 or -NR 15 (CO)R 16 .

[0095] Embodiment 22 provides a method according to any one of embodiments 14-19, wherein R 6 is hydrogen, chlorine, fluorine, methyl, ethyl, difluoromethyl, hydroxy, methoxy, ethoxy, (methoxy)methyl, (ethoxy)methyl, (methoxy)ethyl, (ethoxy)ethyl, oxetanyl, tetrahydrofuranyl, -C(O)-difluoromethyl, -NH 2 or -NH(CO)CH 3 .

[0096] Embodiment 23 provides a method according to any one of embodiments 14-19, wherein R 6 Halogen, C 1 -C 6 Alkyl, C 1 -C 6 Haloalkyl, hydroxyl, C 1 -C 6 Alkoxy, C 1 -C 3 Alkoxy C 1 -C 3 Alkyl, C 3 -C 6 Heterocycloalkyl, -C(O)-C 1 -C 3 Haloalkyl, -N(R 9 ) 2 or -NR 15 (CO)R 16 .

[0097] Embodiment 24 provides a method according to any one of embodiments 14-19, wherein R 6Halogen, C 1 -C 3 Alkyl, C 1 -C 3 Haloalkyl, hydroxyl, C 1 -C 3 Alkoxy, C 1 -C 3 Alkoxy C 1 -C 3 Alkyl, C 3 -C 6 Heterocycloalkyl, -C(O)-C 1 -C 3 Haloalkyl, -N(R 9 ) 2 or -NR 15 (CO)R 16 .

[0098] Embodiment 25 provides a method according to any one of embodiments 14-19, wherein 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 2 or -NH(CO)CH 3 .

[0099] Embodiment 26 provides a method according to any one of embodiments 23-25, wherein G, Q, J and U are each independently C(H).

[0100] Embodiment 27 provides a method according to any one of embodiments 23-25, wherein G, Q, J and U are independently selected from C(H) and C(R 5 ).

[0101] Embodiment 28 provides a method according to any one of embodiments 23-25, wherein G, Q, J and U are independently selected from C(H) and N.

[0102] Embodiment 29 provides a method according to any one of embodiments 14-19, wherein

[0103] R 6 It is hydrogen;

[0104] At least one of G, Q, J and U is C(R 5 ), and the remaining G, Q, J and U are independently selected from C(H), C(R 5 ) and N, where R 5 are independently hydroxy, halogen, C 1 -C 6 Alkyl, C1 -C 6 Haloalkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkoxy, C 3 -C 6 Cycloalkoxy, C 3 -C 6 Cycloalkyl, C 3 -C 6 Heterocycloalkyl or C 1 -C 3 Alkoxy C 1 -C 3 alkyl.

[0105] Embodiment 30 provides a method according to embodiment 29, wherein one or two of G, Q, J and U are N.

[0106] Embodiment 31 provides a method according to any one of embodiments 14-19, wherein

[0107] R 6 It is hydrogen;

[0108] At least one of G, Q, J and U is C(R 5 ), and the remaining G, Q, J and U are independently selected from C(H) and C(R 5 ), where R 5 are independently hydroxy, halogen, C 1 -C 6 Alkyl, C 1 -C 6 Haloalkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkoxy, C 3 -C 6 Cycloalkoxy, C 3 -C 6 Cycloalkyl, C 3 -C 6 Heterocycloalkyl or C 1 -C 3 Alkoxy C 1 -C 3 alkyl.

[0109] Embodiment 32 provides a method according to embodiment 31, wherein 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 ).

[0110] Embodiment 33 provides a method according to embodiment 31, wherein two of G, Q, J and U are C(R 5 ), and the remaining G, Q, J and U are independently C(H).

[0111] Embodiment 34 provides a method according to embodiment 31, wherein three of G, Q, J and U are C(R 5 ), and the remaining G, Q, J and U are C(H).

[0112] Embodiment 35 provides a method according to any one of embodiments 14-19, wherein G, Q, J and U together with the thiophene to which they are attached form:

[0113]

[0114] Embodiment 36 provides a method according to embodiment 35, wherein G, Q, J and U together with the thiophene ring to which they are attached form benzo[b]thiophene.

[0115] Embodiment 37 provides a method according to any one of embodiments 14-36, wherein R 5 , if present, is hydroxyl, halogen, C 1 -C 3 Alkyl, C 1 -C 3 Haloalkyl, C 1 -C 3 Alkoxy, C 1 -C 3 Haloalkoxy, C 3 -C 6 Cycloalkoxy, C 3 -C 6 Cycloalkyl, C 3 -C 6 Heterocycloalkyl or C 1 -C 3 Alkoxy C 1 -C 3 alkyl.

[0116] Embodiment 38 provides a method according to any one of embodiments 14-36, wherein R 5 , if present, is hydroxyl, halogen, C 1 -C 3 Alkyl, C 1 -C 3 Haloalkyl, C 1 -C 3 Alkoxy, C 1 -C 3 Haloalkoxy, C 3 -C 6Heterocycloalkyl or C 1 -C 3 Alkoxy C 1 -C 3 alkyl.

[0117] Embodiment 39 provides a method according to any one of embodiments 14-36, wherein R 5 , if present, is hydroxy, chloro, fluoro, methyl, ethyl, methoxy, ethoxy, 2,2-difluoroethoxy, oxetanyl, tetrahydrofuranyl, (methoxy)methyl, (ethoxy)methyl, (methoxy)ethyl or (ethoxy)ethyl.

[0118] Embodiment 40 provides a method according to any one of embodiments 14-39, wherein R 7 It's methyl.

[0119] Embodiment 41 provides a method according to any one of embodiments 14-39, wherein R 7 It is ethyl.

[0120] Embodiment 42 provides a method according to any one of embodiments 14-39, wherein R 7 is propyl (eg, isopropyl).

[0121] Embodiment 43 provides a method according to any one of embodiments 14-39, wherein R 7 It is difluoromethyl or trifluoromethyl.

[0122] Embodiment 44 provides a method according to embodiment 14, wherein the PRMT5 inhibitor has the formula:

[0123]

[0124] G, Q, J and U together with the thiophene to which they are attached form:

[0125]

[0126] Where R 5 are independently hydroxy, halogen, C 1 -C 3 Alkyl, C 1 -C 3 Haloalkyl, C 1 -C 3 Alkoxy, C 1 -C 3 Haloalkoxy, C 3 -C 6 Heterocycloalkyl or C 1 -C 3 Alkoxy C 1 -C 3alkyl; and

[0127] R 6 is hydrogen, halogen, C 1 -C 3 Alkyl, C 1 -C 3 Haloalkyl, hydroxyl, C 1 -C 3 Alkoxy, C 1 -C 3 Alkoxy C 1 -C 3 Alkyl, C 3 -C 6 Heterocycloalkyl, -C(O)-C 1 -C 3 Haloalkyl, -N(R 9 ) 2 or -NR 15 (CO)R 16 .

[0128] Embodiment 45 provides a method according to embodiment 14, wherein the PRMT5 inhibitor has the formula:

[0129]

[0130] in

[0131] G, Q, J and U together with the thiophene to which they are attached form:

[0132]

[0133] Where R 5 are independently hydroxy, halogen, C 1 -C 3 Alkyl, C 1 -C 3 Haloalkyl, C 1 -C 3 Alkoxy, C 1 -C 3 Halogenated alkoxy, C 3 -C 6 Heterocycloalkyl or C 1 -C 3 Alkoxy C 1 -C 3 alkyl; and

[0134] R 6 Halogen, C 1 -C 3 Alkyl, C 1 -C 3 Haloalkyl, hydroxyl, C 1 -C 3Alkoxy, C 1 -C 3 Alkoxy C 1 -C 3 Alkyl, C 3 -C 6 Heterocycloalkyl, -C(O)-C 1 -C 3 Haloalkyl, -N(R 9 ) 2 or -NR 15 (CO)R 16 .

[0135] Embodiment 46 provides a method according to embodiment 14, wherein the PRMT5 inhibitor has the formula:

[0136]

[0137] in

[0138] G, Q, J and U together with the thiophene to which they are attached form:

[0139]

[0140] Where R 5 are independently hydroxy, halogen, C 1 -C 3 Alkyl, C 1 -C 3 Haloalkyl, C 1 -C 3 Alkoxy, C 1 -C 3 Halogenated alkoxy, C 3 -C 6 Heterocycloalkyl or C 1 -C 3 Alkoxy C 1 -C 3 alkyl.

[0141] Embodiment 47 provides the method of the present disclosure, wherein the PRMT5 inhibitor is a compound of formula (IIIB):

[0142]

[0143] or a pharmaceutically acceptable salt thereof, wherein

[0144] A is for CR 9 or N;

[0145] D is -CH 2 -NH 2 ,

[0146] W is CR 9 or N, where R 9 Is H or C 1 -C 3 alkyl;

[0147] R 51 is hydrogen, fluorine, chlorine or methyl, or R 51 and R 52 Together with the atoms to which it is attached, it forms C 4 -C 6 Heterocycloalkyl (e.g., hydrofuranyl);

[0148] R 52 is fluorine, chlorine or methyl, or R 52 and R 53 Together with the atom to which it is attached, it forms a phenyl group;

[0149] R 53 is hydrogen, fluorine, chlorine or methyl;

[0150] R 54 It is hydrogen, halogen, C 1 -C 3 Alkyl or C 1 -C 3 Alkoxy;

[0151] L 5 Is -O- or -CH 2 -;

[0152] R 6 It is hydrogen, halogen, C 1 -C 6 Alkyl, hydroxyl, C 1 -C 6 Alkoxy, C 1 -C 3 Alkoxy C 1 -C 3 Alkyl, C 3 -C 6 Heterocycloalkyl, -C(O)-C 1 -C 3 Haloalkyl or -NR 15 (CO)R 16 , where R 15 is hydrogen or methyl, and R 16 It is C 1 -C 3 alkyl;

[0153] R 7 It is C 1 -C 3 Alkyl or C 1 -C 3 Halogenated alkyl.

[0154] Embodiment 48 provides a method according to embodiment 47, wherein:

[0155] A is -CH or -CCH 3 ;

[0156] D is -CH 2 -NH 2 ;

[0157] W is -CH, -CCH 3 or N;

[0158] R 51 , R 52 , R 53 and R 54 are each independently selected from hydrogen, fluorine, chlorine or methyl;

[0159] L 5 is-O-;

[0160] R 6 is hydrogen, fluorine, chlorine or methyl; and

[0161] R 7 It is C 1 -C 2 Alkyl or C 1 -C 2 Halogenated alkyl.

[0162] Embodiment 49 provides a method according to embodiment 47 or embodiment 48, wherein:

[0163] A and W are -CH;

[0164] D is -CH 2 -NH 2 ;

[0165] R 51 , R 52 and R 53 are each independently selected from hydrogen, fluorine, chlorine and methyl;

[0166] R 54 It is hydrogen;

[0167] L 5 is-O-;

[0168] R 6 is hydrogen; and

[0169] R 7 It's methyl.

[0170] Embodiment 50 provides a method according to any one of embodiments 47-49, wherein:

[0171] A and W are -CH;

[0172] D is -CH 2 -NH 2 ;

[0173] R 51 and R 52 are each independently selected from fluorine, chlorine and methyl;

[0174] R 53 and R 54 It is hydrogen;

[0175] L 5 is-O-;

[0176] R 6 is hydrogen; and

[0177] R 7 It's methyl.

[0178] Embodiment 51 provides a method according to embodiment 47, wherein A is CH.

[0179] Embodiment 52 provides a method according to embodiment 47 or 48, wherein W is N.

[0180] Embodiment 53 provides a method according to embodiment 47 or 48, wherein W is CH.

[0181] Embodiment 54 provides a method according to any one of embodiments 47-50, wherein D is -CH 2 -NH 2 .

[0182] Embodiment 55 provides a method according to any one of embodiments 47-51, wherein R 54 is hydrogen or methyl.

[0183] Embodiment 56 provides a method according to any one of embodiments 47-51, wherein R 54 It's hydrogen.

[0184] Embodiment 57 provides a method according to any one of embodiments 47-51, wherein R 54 It's methyl.

[0185] Embodiment 58 provides a method according to embodiment 47, wherein the PRMT5 inhibitor has the formula:

[0186] For example,

[0187] Embodiment 59 provides a method according to any one of embodiments 47-55, wherein L 5Yes-CH 2 -.

[0188] Embodiment 60 provides a method according to any one of embodiments 47-55, wherein L 5 Yes -O-.

[0189] Embodiment 61 provides a method according to any one of embodiments 47-57, wherein R 6 It is hydrogen, halogen, C 1 -C 3 Alkyl, C 1 -C 3 Haloalkyl, hydroxyl, C 1 -C 3 Alkoxy, C 1 -C 3 Alkoxy C 1 -C 3 Alkyl, C 3 -C 6 Heterocycloalkyl, -C(O)-C 1 -C 3 Haloalkyl, -N(R 9 ) 2 or -NR 15 (CO)R 16 ; For example, where R 6 is hydrogen, chlorine, fluorine, methyl, ethyl, difluoromethyl, hydroxy, methoxy, ethoxy, (methoxy)methyl, (ethoxy)methyl, (methoxy)ethyl, (ethoxy)ethyl, oxetanyl, tetrahydrofuranyl, -C(O)-difluoromethyl, -NH 2 or -NH(CO)CH 3 .

[0190] Embodiment 62 provides a method according to any one of embodiments 47-57, wherein R 6 It is hydrogen, halogen, C 1 -C 6 Alkyl or C 1 -C 6 Alkoxy; for example, R 6 It is hydrogen, halogen, C 1 -C 3 Alkyl or C 1 -C 3 Alkoxy.

[0191] Embodiment 63 provides a method according to any one of embodiments 47-57, wherein R 6 is hydrogen, chlorine, fluorine, methyl, ethyl, methoxy or ethoxy.

[0192] Embodiment 64 provides a method according to any one of embodiments 47-57, wherein R6 Halogen, C 1 -C 3 Alkyl, C 1 -C 3 Haloalkyl, hydroxyl, C 1 -C 3 Alkoxy, C 1 -C 3 Alkoxy C 1 -C 3 Alkyl, C 3 -C 6 Heterocycloalkyl, -C(O)-C 1 -C 3 Haloalkyl, -N(R 9 ) 2 or -NR 15 (CO)R 16 ; For example, where 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 2 or -NH(CO)CH 3 .

[0193] Embodiment 65 provides a method according to any one of embodiments 47-57, wherein R 6 Halogen, C 1 -C 6 Alkyl or C 1 -C 6 Alkoxy; for example, R 6 Halogen, C 1 -C 3 Alkyl or C 1 -C 3 Alkoxy.

[0194] Embodiment 66 provides a method according to any one of embodiments 47-57, wherein R 6 is chlorine, fluorine, methyl, ethyl, methoxy or ethoxy.

[0195] Embodiment 67 provides a method according to any one of embodiments 47-63, wherein R 7 It's methyl.

[0196] Embodiment 68 provides a method according to any one of embodiments 47-63, wherein R 7 It is ethyl.

[0197] Embodiment 69 provides a method according to any one of embodiments 47-63, wherein R 7is propyl (eg, isopropyl).

[0198] Embodiment 70 provides a method according to any one of embodiments 47-63, wherein R 7 It is difluoromethyl or trifluoromethyl.

[0199] Embodiment 71 provides a method according to any one of embodiments 47-67, wherein R 53 is hydrogen or methoxy; or wherein R 53 It's hydrogen.

[0200] Embodiment 72 provides a method according to embodiment 47, wherein the PRMT5 inhibitor has the formula:

[0201]

[0202] Embodiment 73 provides a method according to any one of embodiments 47-69, wherein R 52 is fluorine, and R 51 is hydrogen, fluorine, chlorine or methyl.

[0203] Embodiment 74 provides a method according to any one of embodiments 47-69, wherein R 52 is fluorine, and R 51 It's chlorine.

[0204] Embodiment 75 provides a method according to any one of embodiments 47-69, wherein R 52 is fluorine, and R 51 is methyl or hydrogen (e.g., R 52 is fluorine and R 51 is methyl; or R 52 is fluorine and R 51 is hydrogen).

[0205] Embodiment 76 provides a method according to any one of embodiments 47-69, wherein R 51 and R 52 Together with the atoms to which they are attached, they form a hydrofuranyl group (e.g., ).

[0206] Embodiment 77 provides a method according to any one of embodiments 47-76, wherein the PRMT5 inhibitor is

[0207]

[0208] Embodiment 78 provides a method according to any one of embodiments 47-77, wherein the PRMT5 inhibitor is

[0209]

[0210] One invention of the present disclosure provides a method, wherein the PRMT5 inhibitor is a compound of formula (IIIA) (Embodiment 79):

[0211]

[0212] or a pharmaceutically acceptable salt thereof, wherein

[0213] A is for CR 9 or N;

[0214] D is -CH 2 -NH 2 ,

[0215] W is CR 9 or N, where R 9 Is H or C 1 -C 3 alkyl;

[0216] R 2 yes

[0217]

[0218] Where R 56 is hydrogen, fluorine, chlorine or methyl,

[0219] G, Q, J and U are independently selected from C(H), C(R 5 ) and N, provided that only one or two of G, Q, J and U may be N;

[0220] R 5 are independently hydroxy, halogen, C 1 -C 6 Alkyl, C 1 -C 6 Haloalkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Halogenated alkoxy, C 3 -C 6 Cycloalkoxy, C 3 -C 6 Cycloalkyl, C 3 -C 6 Heterocycloalkyl or C 1 -C 3 Alkoxy C 1 -C 3 alkyl;

[0221] R 6 is hydrogen, halogen, C 1 -C 6Alkyl, hydroxyl, C 1 -C 6 Alkoxy, C 1 -C 3 Alkoxy C 1 -C 3 Alkyl, C 3 -C 6 Heterocycloalkyl, -C(O)-C 1 -C 3 Haloalkyl or -NR 15 (CO)R 16 , where R 15 is hydrogen or methyl, and R 16 It is C 1 -C 3 alkyl; and

[0222] R 7 It is C 1 -C 3 Alkyl or C 1 -C 3 Halogenated alkyl.

[0223] One invention of the present disclosure provides a method, wherein the PRMT5 inhibitor is a compound of formula (IIIA) (embodiment 80):

[0224]

[0225] or a pharmaceutically acceptable salt thereof, wherein

[0226] A is for CR 9 or N;

[0227] D is -CH 2 -NH 2 ,

[0228] W is CR 9 or N, where R 9 Is H or C 1 -C 3 alkyl;

[0229] R 2 yes

[0230]

[0231] Where R 56 is hydrogen, halogen, C 1 -C 6 Alkyl, C 1 -C 6 Haloalkyl, C 1 -C 6 Alkoxy or C 1 -C6 Haloalkoxy;

[0232] R 6 is hydrogen, halogen, C 1 -C 6 Alkyl, hydroxyl, C 1 -C 6 Alkoxy, C 1 -C 3 Alkoxy C 1 -C 3 Alkyl, C 3 -C 6 Heterocycloalkyl, -C(O)-C 1 -C 3 Haloalkyl or -NR 15 (CO)R 16 , where R 15 is hydrogen or methyl, and R 16 It is C 1 -C 3 alkyl; and

[0233] R 7 It is C 1 -C 3 Alkyl or C 1 -C 3 Halogenated alkyl.

[0234] Embodiment 81 provides a method according to embodiment 79 or 80, wherein A is CH.

[0235] Embodiment 82 provides a method according to embodiment 79 or 80, wherein W is N.

[0236] Embodiment 83 provides a method according to embodiment 79 or 80, wherein W is CH.

[0237] Embodiment 84 provides a method according to any one of embodiments 79 or 80, wherein D is -CH 2 -NH 2 .

[0238] Embodiment 85 provides a method according to embodiment 79 or 80, which has the formula:

[0239]

[0240] Embodiment 86 provides a method according to Embodiment 79 or 81-85, wherein R 2 yes

[0241]

[0242] Embodiment 87 provides a method according to embodiment 86, wherein G, Q, J and U are independently selected from C(H) and C(R 5 ).

[0243] Embodiment 88 provides a method according to embodiment 86, wherein G, Q, J and U are independently C(H).

[0244] Embodiment 89 provides a method according to embodiment 86, wherein 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 ).

[0245] Embodiment 90 provides a method according to embodiment 86, wherein U is N, and G, Q and J are independently selected from C(H) and C(R 5 ).

[0246] Embodiment 91 provides a method according to embodiment 86, wherein G is N, and Q, J and U are independently selected from C(H) and C(R 5 ).

[0247] Embodiment 92 provides a method according to any one of embodiments 79 or 81-91, wherein R 5 , if present, is hydroxyl, halogen, C 1 -C 3 Alkyl, C 1 -C 3 Haloalkyl, C 1 -C 3 Alkoxy, C 1 -C 3 Haloalkoxy, C 3 -C 6 Cycloalkoxy, C 3 -C 6 Cycloalkyl, C 3 -C 6 Heterocycloalkyl or C 1 -C 3 Alkoxy C 1 -C 3 alkyl.

[0248] Embodiment 93 provides a method according to any one of embodiments 79 or 81-91, wherein R 5 , if present, is hydroxyl, halogen, C 1 -C 3 Alkyl, C 1 -C 3 Haloalkyl, C 1 -C 3 Alkoxy, C1 -C 3 Halogenated alkoxy, C 3 -C 6 Heterocycloalkyl or C 1 -C 3 Alkoxy C 1 -C 3 alkyl.

[0249] Embodiment 94 provides a method according to any one of embodiments 79 or 81-91, wherein R 5 , if present, is hydroxy, chloro, fluoro, methyl, ethyl, methoxy, ethoxy, 2,2-difluoroethoxy, oxetanyl, tetrahydrofuranyl, (methoxy)methyl, (ethoxy)methyl, (methoxy)ethyl or (ethoxy)ethyl.

[0250] Embodiment 95 provides a method according to any one of embodiments 79 or 81-91, wherein R 5 , if present, is a halogen, C 1 -C 6 Alkyl or C 1 -C 6 Alkoxy; for example, R 6 Halogen, C 1 -C 3 Alkyl or C 1 -C 3 Alkoxy.

[0251] Embodiment 96 provides a method according to any one of embodiments 79 or 81-91, wherein R 5 , if present, is chloro, fluoro, methyl, ethyl, methoxy or ethoxy.

[0252] Embodiment 97 provides a method according to any one of embodiments 79 or 81-91, wherein R 56 is fluorine, chlorine or methyl.

[0253] Embodiment 98 provides the method according to embodiments 80-85, wherein R 2 yes

[0254]

[0255] Embodiment 99 provides a method according to any one of embodiments 80-85 or 98, wherein R 56 is hydrogen, fluorine, chlorine or methyl.

[0256] Embodiment 100 provides a method according to any one of embodiments 79-99, wherein R 6 is hydrogen, halogen, C 1 -C 3 Alkyl, C1 -C 3 Haloalkyl, hydroxyl, C 1 -C 3 Alkoxy, C 1 -C 3 Alkoxy C 1 -C 3 Alkyl, C 3 -C 6 Heterocycloalkyl, -C(O)-C 1 -C 3 Haloalkyl, -N(R 9 ) 2 or -NR 15 (CO)R 16 ; For example, where R 6 is hydrogen, chlorine, fluorine, methyl, ethyl, difluoromethyl, hydroxy, methoxy, ethoxy, (methoxy)methyl, (ethoxy)methyl, (methoxy)ethyl, (ethoxy)ethyl, oxetanyl, tetrahydrofuranyl, -C(O)-difluoromethyl, -NH 2 or -NH(CO)CH 3 .

[0257] Embodiment 101 provides a method according to any one of embodiments 79-99, wherein R 6 It is hydrogen, halogen, C 1 -C 6 Alkyl or C 1 -C 6 Alkoxy; for example, R 6 It is hydrogen, halogen, C 1 -C 3 Alkyl or C 1 -C 3 Alkoxy.

[0258] Embodiment 102 provides a method according to any one of Embodiments 79-99, wherein R 6 is hydrogen, chlorine, fluorine, methyl, ethyl, methoxy or ethoxy.

[0259] Embodiment 103 provides a method according to any one of embodiments 79-99, wherein R 6 Halogen, C 1 -C 3 Alkyl, C 1 -C 3 Haloalkyl, hydroxyl, C 1 -C 3 Alkoxy, C 1 -C 3 Alkoxy C 1 -C 3 Alkyl, C 3 -C6 Heterocycloalkyl, -C(O)-C 1 -C 3 Haloalkyl, -N(R 9 ) 2 or -NR 15 (CO)R 16 ; For example, where 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 2 or -NH(CO)CH 3 .

[0260] Embodiment 104 provides a method according to any one of embodiments 79-99, wherein R 6 Halogen, C 1 -C 6 Alkyl or C 1 -C 6 Alkoxy; for example, R 6 Halogen, C 1 -C 3 Alkyl or C 1 -C 3 Alkoxy.

[0261] Embodiment 105 provides a method according to any one of embodiments 79-99, wherein R 6 is chlorine, fluorine, methyl, ethyl, methoxy or ethoxy.

[0262] Embodiment 106 provides a method according to any one of embodiments 79-105, wherein R 7 It's methyl.

[0263] Embodiment 107 provides a method according to any one of embodiments 79-105, wherein R 7 It is ethyl.

[0264] Embodiment 108 provides a method according to any one of embodiments 79-105, wherein R 7 is propyl (eg, isopropyl).

[0265] Embodiment 109 provides a method according to any one of embodiments 79-105, wherein R 7 It is difluoromethyl or trifluoromethyl.

[0266] In certain embodiments of the methods of the present disclosure as described herein, the PRMT5 inhibitor is:

[0267]

[0268] In certain embodiments of the methods of the present disclosure as described herein, the PRMT5 inhibitor is:

[0269]

[0270]

[0271] In certain embodiments of the methods of the present disclosure as described herein, the PRMT5 inhibitor is:

[0272]

[0273] In certain embodiments of the methods of the present disclosure as described herein, the PRMT5 inhibitor is:

[0274]

[0275] In one aspect, the present disclosure provides a method of treating cancer in an individual, the method comprising administering to the individual:

[0276] A therapeutically effective amount of ABT-199 (venetoclax), wherein ABT-199 is:

[0277]

[0278] A therapeutically effective amount of a PRMT5 inhibitor of the formula:

[0279]

[0280] In one aspect, the present disclosure provides a method of treating cancer in an individual, the method comprising administering to the individual:

[0281] A therapeutically effective amount of ABT-263 (Navitoclax), wherein ABT-263 is:

[0282] and

[0283] A therapeutically effective amount of a PRMT5 inhibitor of the formula:

[0284]

[0285] The PRMT5 inhibitors and / or BCL-2 family inhibitors disclosed herein (e.g., ABT-199 (venetoclax), ABT-263 (navitoc), A-1155463, A-1331852, obaclax (GX15-070), ABT-737, TW-37, gossypol or (R)-(-)-gossypol, HA14-1, sabutoclax and / or DT2216) can be provided as a pharmaceutical composition comprising a therapeutically effective amount of such inhibitors and a pharmaceutically acceptable carrier, excipient and / or diluent. The PRMT5 inhibitors and / or BCL-2 family inhibitors disclosed herein can be formulated by any method known in the art and can be prepared for administration by any route, including but not limited to parenteral, oral, sublingual, transdermal, topical, intranasal, intratracheal or rectal.

[0286] The characteristics of the carrier will depend on the route of administration. The term "pharmaceutically acceptable" as used herein refers to a non-toxic substance that is compatible with a biological system such as a cell, cell culture, tissue or organism, and does not interfere with the effectiveness of the biological activity of the active ingredient. Therefore, in addition to the inhibitor, the pharmaceutical composition of the present disclosure may contain diluents, fillers, salts, buffers, stabilizers, solubilizers and other materials well known in the art. The preparation of pharmaceutically acceptable preparations is described in, for example, Remington's Pharmaceutical Sciences, 18th edition, ed. A. Gennaro, Mack Publishing Co., Easton, Pa., 1990.

[0287] The PRMT5 inhibitors and BCL-2 family inhibitors disclosed herein are administered in a therapeutically effective amount. As used herein, the phrase "therapeutically effective amount" or "effective amount" refers to the amount of an active agent that causes a biological or medical response sought by a researcher, physician or other clinician in a tissue, system, individual or person. Typically, a therapeutically effective amount is sufficient to deliver a biological or drug response to an individual without causing severe toxic effects. The dosage range of the active agent may be about 1 to 500 mg / m per day. 2 , such as 5 to 400 mg / m per day 2 , more usually 10 to 300 mg / m per day 2 Recipient body weight. A typical topical dosage is 0.01 to 10% wt / wt in a suitable carrier.

[0288] In certain embodiments of the methods of the present disclosure, the therapeutically effective amount of the PRMT5 inhibitor ranges from about 0.01 to 300 mg / kg / day. For example, in certain embodiments, the therapeutically effective amount of the PRMT5 inhibitor ranges from about 0.1 to 100 mg / kg / day or 25 to 100 mg / kg / day or 50 to 100 mg / kg / day.

[0289] In certain embodiments, the therapeutically effective amount of a PRMT5 inhibitor is less than 1%, such as less than 10% or less than 25% or less than 50% of the clinically established therapeutic amount (the amount of the inhibitor, such as when PRMT5 is administered alone).

[0290] In certain embodiments of the methods of the present disclosure, the therapeutically effective amount of the BCL-2 family inhibitor ranges from about 1 to 500 mg / m per day. 2 , such as 5 to 400 mg / m per day 2 , more usually 10 to 300 mg / m per day 2 For 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 (eg, 50 to 250 mg / m 2 or 50 to 200 mg / m 2 or 50 to 150 mg / m 2 ).

[0291] For example, in various embodiments, the BCL-2 family inhibitor can be ABT-199 (venetoclax), ABT-263 (navitoc), A-1155463, A-1331852, obaclax (GX15-070), ABT-737, TW-37, gossypol or (R)-(-)-gossypol, HA14-1, sabutoclax or DT2216. Thus, in certain embodiments of the methods of the present disclosure, the therapeutically effective amount of each of these active agents ranges from about 1 to 500 mg / m per day. 2 , such as 5 to 400 mg / m per day 2 , more usually 1 0 to 300 mg / m 2 For example, in certain embodiments, a therapeutically effective amount of any of these active agents ranges from about 30 to 300 mg / m per day. 2 (eg, 50 to 250 mg / m 2 or 50 to 200 mg / m 2 or 50 to 150 mg / m 2 ).

[0292] In certain embodiments, the therapeutically effective amount of ABT-199 (venetoclax), ABT-263 (navitoc), A-1155463, A-1331852, obaclax (GX15-070), ABT-737, TW-37, gossypol or (R)-(-)-gossypol, HA14-1, sabutoclax, or DT2216 is less than 1%, e.g., less than 10% or less than 25% or less than 50% or less than 75% of the clinically established therapeutic amount (e.g., the amount required when the compound is administered by itself).

[0293] In defining the uses of the PRMT5 inhibitors and BCL-2 family inhibitors of the present disclosure (e.g., ABT-199 (venetoclax), ABT-263 (navitoc), A-1155463, A-1331852, obaclax (GX15-070), ABT-737, TW-37, gossypol or (R)-(-)-gossypol, HA14-1, sabutoclax or DT2216), the combination therapy is intended to include the administration of each active agent in a sequential manner (e.g., the PRMT5 inhibitors and BCL-2 family inhibitors of the present disclosure can be formulated as separate compositions that are administered sequentially) in a regimen that will provide the beneficial effects of the drug combination, and is also intended to include the co-administration of these active agents in a substantially simultaneous manner (e.g., in a single dosage form having a fixed ratio of these active agents or in multiple or separate individual dosage forms of each active agent). The present disclosure is not limited by the order of administration: the PRMT5 inhibitors of the present disclosure may be administered before or after (ie, sequentially) or at the same time (ie, simultaneously) as the BCL-2 family inhibitors of the present disclosure.

[0294] The methods of the present disclosure may be used as a first-line treatment. Thus, in certain embodiments of the methods of the present disclosure, the individual has not previously received another first-line therapy.

[0295] The method of the present disclosure can also be used as a first-line maintenance or second-line treatment. Therefore, in certain embodiments of the method of the present disclosure, the individual has previously completed another first-line therapy. For example, in certain embodiments, the method of the present disclosure can provide a delay to cancer progression and recurrence in an individual who has previously completed another first-line chemotherapy. For example, in certain embodiments, the individual has previously completed a treatment regimen, which may include but is not limited to chemotherapy, targeted therapy and immunotherapy, as a single active agent or in combination with other therapies. In certain embodiments of the method of the present disclosure, the individual has previously completed another first-line chemotherapy and is partially responsive to such chemotherapy.

[0296] definition

[0297] For simplicity, chemical groups are primarily defined and referred to throughout the text as monovalent chemical groups (e.g., alkyl, aryl, etc.). However, under appropriate structural circumstances that are clear to those skilled in the art, these terms may also be used to refer to corresponding multivalent groups. For example, although an "alkyl" group generally refers to a monovalent group (e.g., CH 3 -CH 2 -), but in some cases the divalent linking group may be an "alkyl" group, in which case one skilled in the art would understand that an alkyl group is a divalent group (e.g., -CH 2 -CH 2 -), which is equivalent to the term "alkylene". Similarly, where a divalent group is required and described as "aryl", those skilled in the art will understand that the term "aryl" refers to the corresponding divalent group, arylene. All atoms are understood to have a normal number of their valences for bond formation (i.e., 4 for carbon, 3 for N, 2 for O, and 2, 4 or 6 for S, depending on the oxidation state of S).

[0298] The term "amino" refers to -NH 2 .

[0299] The term "acetyl" refers to "-C(O)CH 3 .

[0300] As used herein, the term "acyl" refers to an alkylcarbonyl or arylcarbonyl substituent wherein the alkyl and aryl portions are as defined herein.

[0301] As used herein, the term "alkyl" refers to saturated straight-chain and branched-chain aliphatic groups having 1 to 12 carbon atoms. Thus, "alkyl" includes C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , C 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.

[0302] As used herein, the term "alkenyl" refers to an unsaturated straight or branched chain aliphatic group having one or more carbon-carbon double bonds, having 2 to 12 carbon atoms. Thus, "alkenyl" includes C 2 , C 3 , C 4 , C 5 , C 6 , C7 , C 8 , C 9 , C 10 , C 11 and C 12 Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, and hexenyl.

[0303] As used herein, the term "alkynyl" refers to an unsaturated straight or branched chain aliphatic group having one or more carbon-carbon triple bonds and having 2 to 12 carbon atoms. 2 , C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , C 10 , C 11 and C 12 Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, and hexynyl.

[0304] As defined above, an "alkylene," "alkenylene," or "alkynylene" group is an alkyl, alkenyl, or alkynyl group that is positioned between and serves to connect 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.

[0305] The term "alkoxy" refers to -OC 1 -C 6 alkyl.

[0306] As used herein, the term "cycloalkyl" is a saturated and partially unsaturated cyclic hydrocarbon group having 3 to 12 carbon atoms. Thus, "cycloalkyl" includes C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , C 10 , C 11 and C 12 Cyclic hydrocarbon groups. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.

[0307] The term "heteroalkyl" refers to an alkyl group as defined above in which one or more carbon atoms in the chain are independently replaced by O, S or NRx Substitute, where R x is hydrogen or C 1 -C 3 Examples of heteroalkyl groups include methoxymethyl, methoxyethyl and methoxypropyl.

[0308] "Aryl" is a C 6 -C 14 Aromatic moieties. Thus, "aryl" includes C 6 , C 10 , C 13 and C 14 Exemplary aryl groups are C 6 -C 10 Aryl. Specific aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, and fluorenyl. "Aryl" 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, such as indenyl.

[0309] "Aralkyl" or "arylalkyl" includes an aryl group covalently linked to an alkyl group, wherein the group is linked to another group through the alkyl group. Exemplary aralkyl groups are -(C 1 -C 6 )alkyl(C 6 -C 10 ) aryl groups, including but not limited to benzyl, phenethyl and naphthylmethyl. 1 -C 3 Alkyl is C 1 -C 3 An alkyl group is covalently linked to an aryl group.

[0310] A "heterocyclyl" or "heterocyclic" group is a monocyclic or bicyclic (fused or spiro) ring structure having 3-12 atoms (3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 atoms), for example 4-8 atoms, in which one or more ring atoms are independently -C(O)-, N, NR 4 , O or S, the remaining ring atoms are quaternary carbon or carbonyl carbon.The example of heterocyclic radical includes but is not limited to epoxy, oxiranyl, oxetane, azetidinyl, aziridine, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, piperazinyl, imidazolidinyl, thiazolidinyl, thiatanyl, dithianyl, trithianyl, nitrogen thiohexyl (azathianyl), oxathianyl, dioxolanyl, oxazolidinyl, oxazolidinone, decahydroquinolyl, piperidone, 4-piperidone, thiomorpholinyl, dimethyl-morpholinyl and morpholinyl.Particularly excluded from the scope of this term are compounds with adjacent O and / or S ring atoms.

[0311] As used herein, "L-heterocyclyl" refers to a heterocyclyl group covalently linked to another group via an alkylene linker.

[0312] 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; sharing 6, 10 or 14 pi electrons in a cyclic array; and having, in addition to carbon atoms, one to three heteroatoms each independently being 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 N, O or S ring atoms. Examples of heteroaryl groups include acridinyl, azinyl, benzimidazolyl, benzofuranyl, benzo[d]oxazol-2(3H)-one, 2H-benzo[b][1,4]oxazin-3(4H)-one, benzothiophenyl, benzothiophenyl, benzoxazolyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, furanyl, furazanyl, 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, oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiyl, phenoxazinyl, phthalazinyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolidinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridinyl, pyrimidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinine cycloalkyl, tetrahydroisoquinolyl, tetrahydroquinolyl, 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, thienooxazolyl, thienoimidazolyl, thienyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl and xanthenyl.

[0313] "L-heteroaralkyl" or "L-heteroarylalkyl" includes a heteroaryl group covalently linked to another group through an alkylene linker. Examples of heteroalkyl groups include C 1 -C 6alkyl and heteroaryl having 5, 6, 9 or 10 ring atoms. Examples of heteroaralkyl include pyridylmethyl, pyridylethyl, pyrrolylmethyl, pyrrolylethyl, imidazolylmethyl, imidazolylethyl, thiazolylmethyl, thiazolylethyl, benzimidazolylmethyl, benzimidazolylethyl, quinazolinylmethyl, quinolylmethyl, quinolylethyl, benzofuranylmethyl, indolinylethyl, isoquinolylmethyl, isoindolylmethyl, cinnolinylmethyl and benzothienylethyl. Specifically excluded from the scope of this term are compounds having adjacent O and / or S ring atoms.

[0314] "Arylene", "heteroarylene" or "heterocyclylene" as defined above is a divalent aryl, heteroaryl or heterocyclyl group, respectively, which is positioned between two other chemical groups and serves to connect the two other chemical groups.

[0315] As used herein, when a group (e.g., cycloalkyl, aryl, heteroaryl, heterocyclyl, urea, etc.) is described as "optionally substituted" without explicitly stating the substituents, it means that the group optionally has one to four, preferably one to three, more preferably one or two non-hydrogen substituents.

[0316]

[0046] The term "halogen" or "halo" as used herein refers to chlorine, bromine, fluorine or iodine.

[0317] The term "haloalkyl" refers to an alkyl chain in which one or more hydrogens have been replaced by a halogen. Exemplary haloalkyls are trifluoromethyl, difluoromethyl, fluorochloromethyl, chloromethyl and fluoromethyl.

[0318] The term "hydroxyalkyl" refers to an -alkylene-OH group. Example

[0319] 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 specific methods and compounds described therein.

[0320] Example 1 - In vivo combination studies (LU99 model)

[0321] Immunodeficient female nu / nu mice were implanted with 5x10 6 Tumor volume measurements were collected using calipers and calculated using the formula 0.5 × L × W 2 Determine, where L is the length of each tumor and W is the width of each tumor. When the tumor reaches approximately 125-150mm 3Animals were randomly assigned 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 at designated doses and regimens orally (PO) for 20-34 days. Tumor volume was measured twice a week (n=5 / treatment group). Mean tumor volume and mean standard error were calculated and plotted using GraphPad Prism on each study day.

[0322] 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-cyclopropyloxy-3-fluorobenzonitrile, as disclosed in Examples 16-8 on page 307 of International Patent Publication No. WO 2021 / 050915A1 published on March 18, 2021, which is incorporated by reference in its entirety.

[0323] The BCL-2 family inhibitors ABT-199 (venetoclax) and ABT-263 (nervitoclax) used in this example were administered once daily (QD) at 100 mg / kg and were provided by Selleck Chemicals (Houston, TX).

[0324] The results are Figure 1 and are provided in Table 1.

[0325] Table 1

[0326]

[0327] Combination of MRTX1719 with ABT-199 (venetoclax) and ABT-263 (nervetoclax), respectively, resulted in higher antitumor activity in a LU99 cell line-derived xenograft model compared to either compound alone, as measured by changes in tumor volume over time.

[0328] Example 2 - In vivo combination studies (NCI-H1437 model)

[0329] Example 2 was performed according to the in vivo study method described in Example 1 above using xenografts derived from the NCI-H1437 cell line.

[0330] 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-cyclopropyloxy-3-fluorobenzonitrile, as disclosed in Examples 16-8 on page 307 of International Patent Publication No. WO 2021 / 050915A1, published on March 18, 2021, which is incorporated by reference in its entirety. The BCL-2 family inhibitor ABT-263 (navitoc) was administered at 100 mg / kg once daily (QD) and was provided by Selleck Chemicals (Houston, TX). The BCL-2 family inhibitor A-1331852 was administered once daily (QD) at 25 mg / kg and was synthesized at Wuxi AppTec (Wuhan, China).

[0331] The results are Figure 2 and are provided in Table 2.

[0332] Table 2

[0333]

[0334] The combination of MRTX1719 with ABT-263 (Navitoclax) and A-1331852 resulted in greater antitumor activity in the NCI-H1437 cell line-derived xenograft model compared to either compound alone, as measured by changes in tumor volume over time.

[0335] Example 3 - In vivo combination studies (SW1573 model)

[0336] Example 3 was performed according to the in vivo study method described in Example 1 above in xenografts derived from the SW1573 cell line.

[0337] 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-cyclopropyloxy-3-fluorobenzonitrile, as disclosed in Examples 16-8 on page 307 of International Patent Publication No. WO 2021 / 050915A1, published on March 18, 2021, which is incorporated by reference in its entirety. The BCL-2 family inhibitor ABT-263 (navitoc) was administered at 100 mg / kg once daily (QD) and was provided by Selleck Chemicals (Houston, TX). The BCL-2 family inhibitor A-1331852 was administered once daily (QD) at 25 mg / kg and was synthesized at Wuxi AppTec (Wuhan, China).

[0338] The results are Figure 3 and are provided in Table 3

[0339] Table 3

[0340]

[0341]

[0342] The combination of MRTX1719 with ABT-263 (Navitoclax) and A-1331852 resulted in higher antitumor activity in a SW173 cell line-derived xenograft model compared to either compound alone, as measured by changes in tumor volume over time.

[0343] Example 4 - In vivo combination studies (LU99 model)

[0344] Example 4 was performed according to the in vivo study method described in Example 1 above.

[0345] 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-cyclopropyloxy-3-fluorobenzonitrile, as disclosed in Examples 16-8 on page 307 of International Patent Publication No. WO 2021 / 050915A1, published on March 18, 2021, which is incorporated by reference in its entirety. The BCL-2 family inhibitor A-1331852 was administered at 25 mg / kg twice daily (BID) and was synthesized at Wuxi AppTec (Wuhan, China).

[0346] The results are Figure 4and are provided in Table 4.

[0347] Table 4

[0348]

[0349] The combination of MRTX1719 and A-1331852 resulted in greater antitumor activity in a LU99 cell line-derived xenograft model compared to either compound alone, as measured by changes in tumor volume over time.

[0350] Example 5 - In vivo combination study (NCI-H1650 model)

[0351] Example 5 was performed according to the in vivo study method described in Example 1 above using xenografts derived from the NCI-H1650 cell line.

[0352] 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-cyclopropyloxy-3-fluorobenzonitrile, as disclosed in Examples 16-8 on page 307 of International Patent Publication No. WO 2021 / 050915A1, published on March 18, 2021, which is incorporated by reference in its entirety. The BCL-2 family inhibitor ABT-263 (navitoc) was administered at 100 mg / kg once daily (QD) and was provided by Selleck Chemicals (Houston, TX). The BCL-2 family inhibitor A-1331852 was administered at 25 mg / kg twice daily (BID) and was synthesized at Wuxi AppTec (Wuhan, China).

[0353] The results are Figure 5 and are provided in Table 5.

[0354] Table 5

[0355]

[0356] The combination of MRTX1719 with ABT-263 (Navitoclax) and A-1331852 resulted in greater antitumor activity in the NCI-H1650 cell line-derived xenograft model compared to either compound alone, as measured by changes in tumor volume over time.

[0357] Example 6 - Synergy Scoring, In Vitro Combination Studies

[0358] This example illustrates that the combination of an exemplary PRMT5 inhibitor compound of the type described in WO2021 / 050915 and a BCL-2 family inhibitor synergistically inhibits the growth of MTAP-deficient cancer cell lines.

[0359] A panel of MTAP-deficient cancer cell lines was assembled to determine whether the combination of a BCL-2 family inhibitor with an exemplary PRMT5 inhibitor disclosed herein results in synergistic activity.

[0360] [Assays to determine the synergy score for pairwise combinations of each cell line are performed in triplicate. Additional wells of a 384-well or 96-well plate, as well as a separate 384-well or 96-well control plate for determining baseline luminescence, are seeded with cells of the specific MTAP-deficient cell line in an appropriate growth medium for that cell line, such as RPMI 1640 medium supplemented with 10% FBS and any cell line-specific reagents required for growth. The plates are incubated at 37°C in 5% CO. 2 Incubate overnight in atmosphere.

[0361] Cell-Titer Glo reagent (CTG; Promega Corporation) was added to each designated baseline well and the plate was incubated for 20 minutes at room temperature with shaking.Baseline luminescence was quantified using a BMG Clariostar multimode microplate reader according to the manufacturer's instructions.

[0362] A series of 1000X drug dilutions in 100% DMSO was prepared, including 9-point single active 3-fold dilutions of an exemplary PRMT5 inhibitor (of the type described in WO2021 / 050915) with a maximum dose of 3000 nM (and 6-point single active 5-fold dilutions of a BCL-2 family inhibitor with a maximum dose of 3000 nM as a reference standard.

[0363] 10X intermediate dosing plates were prepared in serum-free medium containing arrayed single active agent dilutions of exemplary PRMT5 inhibitors (types described in WO2021 / 050915) and / or BCL-2 family inhibitors. In addition, a matrix of 54 dilution combinations of exemplary PRMT5 inhibitors (types described in WO2021 / 050915) and the BCL-2 family inhibitors was prepared as a test sample.

[0364] Add 10X single active agent and 54 combinations of the dose matrix to each corresponding well of a 384-well plate seeded with the appropriate cell line described above, and incubate the plate at 37 °C in 5% CO. 2The assay was performed incubated for 120 hours in a 4% CO atmosphere. Cell-Titer Glo reagent (CTG) was added to each test well, the plate was incubated at room temperature for 20 minutes with shaking, and the luminescence was quantified using a BMG Clariostar multi-mode microplate reader according to the manufacturer's instructions. In order to standardize the test samples, the baseline luminescence determined was subtracted from each test sample before analysis, and the mean value was calculated from the replicate sample wells.

[0365] The raw data and generated metadata were used as input files to calculate the percent effect of each treatment condition and analyzed using four independent mathematical reference models designed to determine whether two test compounds exhibit synergy: Loewe additivity, Bliss independence, highest single active agent, and ZIP (need reference?).

[0366] The data output from each mathematical model is the assignment of relative synergy scores. The data reported in Table 6 is the summary sum of the Loewe additivity, Bliss independence, highest single active agent, and ZIP synergy scores ("Composite Synergy Score").

[0367] Table 6

[0368] Comprehensive synergy scores of exemplary BCL-2 family inhibitors combined with exemplary PRMT5 inhibitors against MTAP-deficient cancer cell lines

[0369]

[0370] These results demonstrate that a synergistic effect of the combination of multiple BCL-2 family inhibitors with exemplary PRMT5 inhibitor compounds (of the type described in WO2021 / 050915) was observed in each cell line, thereby increasing the sensitivity of MTAP-deficient cell lines to PRMT5 inhibitors.

[0371] It should be understood that the embodiments and implementations described herein are for illustrative purposes only, and those skilled in the art may make various modifications or changes thereto, which should be included within the spirit and scope of the present application and the scope of the appended claims. All publications, patents and patent applications cited herein are incorporated herein by reference.

Claims

1. A method for treating cancer in an individual, the method 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 are administered to the individual.

2. The method of claim 1, wherein the cancer is associated with a homozygous deletion of the methylthioadenosine phosphorylase (MTAP) gene.

3. The method of claim 1 or 2, wherein the PRMT5 inhibitor is a methylthioadenosine (MTA)-cooperative PRMT5 inhibitor.

4. The method of 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, bladder cancer, esophageal cancer, diffuse large B-cell lymphoma, lymphoma, gastric cancer, melanoma, breast cancer, brain cancer, bile duct cancer, mesothelioma, or malignant peripheral nerve sheath tumor.

5. The method of claim 3, wherein the cancer is mesothelioma.

6. The method of claim 3 or 4, wherein the cancer is non-small cell lung cancer.

7. The method of any one of claims 1 to 6, wherein the PRMT5 inhibitor is a compound of Formula IIA, IIB or IIC: or a pharmaceutically acceptable salt thereof, wherein: A is for CR 9 or N; D is (C (R 9 )2) 1-2 -NH2, Or D is wherein the methylene group is bonded to E, wherein E is C; E is C, CR 9 or N; L is each independently a bond or a C1-C3 alkylene group; W is CR 9 or N; X is independently a bond, O, S, -NR 4 -or-NR 4 C(O)-; Z is each independently a bond, -SO-, -SO2-, -CH(OH)-, or -C(O)-; 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 the heterocyclyl, cycloalkyl, aryl and heteroaryl are optionally replaced by one or more R 5 replace; R 4 Each is independently hydrogen or C1-C3 alkyl; R 5 each independently represents 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 by one or more halogens; R 8 is H or C1-C3 alkyl; and R 9 Each is independently H or C1-C3 alkyl, halogen or haloalkyl.

8. The method of any one of claims 1 to 7, wherein the PRMT5 inhibitor is a compound of formula IIIA: or a pharmaceutically acceptable salt thereof, wherein A is for CR 9 or N; D is -CH2-NH2, W is CR 9 or N, where R 9 is H or C1-C3 alkyl; R 2 yes Where R 56 is hydrogen, fluorine, chlorine or methyl, G, Q, J and U are independently selected from C(H), C(R 5 ) and N, provided that only one or two of G, Q, J and U may be N; R 5 Each is independently hydroxy, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C6 cycloalkyloxy, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl or C1-C3 alkoxyC1-C3 alkyl; R 6 is hydrogen, halogen, C1-C6 alkyl, hydroxy, C1-C6 alkoxy, C1-C3 alkoxy C1-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 It is a C1-C3 alkyl group or a C1-C3 haloalkyl group.

9. The method of claim 8, wherein the PRMT5 inhibitor is:

10. The method of any one of claims 1 to 9, wherein the PRMT5 inhibitor is a compound of formula IIIB: or a pharmaceutically acceptable salt thereof, wherein A is for CR 9 or N; D is -CH2-NH2, W is CR 9 or N, where R 9 is H or C1-C3 alkyl; R 51 is hydrogen, fluorine, chlorine or methyl, or R 51 and R 52 Together with the atoms to which it is attached, it forms a C4-C6 heterocycloalkyl group (e.g., hydrofuranyl); R 52 is fluorine, chlorine or methyl, or R 52 and R 53 Together with the atom to which it is attached, it forms a phenyl group; R 53 is hydrogen, fluorine, chlorine or methyl; R 54 is 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 alkoxy C1-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 a C1-C3 alkyl group; R 7 It is a C1-C3 alkyl group or a C1-C3 haloalkyl group.

11. The method of claim 10, wherein: A is -CH or -CCH3; 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, fluorine, chlorine or methyl; L 5 is-O-; R 6 is hydrogen, fluorine, chlorine or methyl; and R 7 It is a C1-C2 alkyl group or a C1-C2 haloalkyl group.

12. The method of claim 10 or claim 11, wherein: A and W are -CH; D is -CH2-NH2; R 51 , R 52 and R 53 are each independently selected from hydrogen, fluorine, chlorine and methyl; R 54 It is hydrogen; L 5 is-O-; R 6 is hydrogen; and R 7 It's methyl.

13. The method of any one of claims 10 to 12, wherein: A and W are -CH; D is -CH2-NH2; R 51 and R 52 are each independently selected from fluorine, chlorine and methyl; R 53 and R 54 It is hydrogen; L 5 is-O-; R 6 is hydrogen; and R 7 It's methyl.

14. The method of claim 10, wherein the PRMT5 inhibitor is: or a pharmaceutically acceptable salt thereof.

15. The method of claim 11, wherein the PRMT5 inhibitor is: or a pharmaceutically acceptable salt thereof.

16. The method of any one of claims 1 to 10, wherein the PRMT5 inhibitor is a compound of formula IIIC: or a pharmaceutically acceptable salt thereof, wherein A is for CR 9 or N; D is -CH2-NH2, W is CR 9 or N, where R 9 is H or C1-C3 alkyl; G, Q, J and U are independently selected from C(H), C(R 5 ) and N, provided that only one or two of G, Q, J and U may be N; R 5 Each is independently hydroxy, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C6 cycloalkyloxy, 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 alkoxy C1-C3 alkyl, C3-C6 heterocycloalkyl, -C(O)-C1-C3 haloalkyl, -N(R 9 )2 or -NR 15 (CO)R 16 , Where R 9 are each independently H or C1-C3 alkyl, R 15 is hydrogen or methyl, and R 16 is C1-C3 alkyl; and R 7 It is a C1-C3 alkyl group or a C1-C3 haloalkyl group.

17. The method of claim 16, wherein the PRMT5 inhibitor is: or a pharmaceutically acceptable salt thereof.

18. The method of any one of claims 1 to 17, wherein the therapeutically effective amount of the PRMT5 inhibitor ranges from about 0.01 to 300 mg / kg / day.

19. The method of any one of claims 1 to 17, wherein the therapeutically effective amount of the PRMT5 inhibitor ranges from about 0.1 to 100 mg / kg / day.

20. The method of any one of claims 1 to 17, wherein the therapeutically effective amount of the PRMT5 inhibitor is less than 1%, e.g., less than 10% or less than 25% or less than 50% of the clinically established therapeutic amount.

21. The method of any one of claims 1-20, wherein the BCL-2 family inhibitor is selected from one or more of the following: ABT-199 (venetoclax), ABT-263 (navitoc), A-1155463, A-1331852, obaclax (GX15-070), ABT-737, TW-37, gossypol, (R)-(-)-gossypol, HA14-1, sabutoclax, and DT2216).

22. The method of any one of claims 1-21, wherein the BCL-2 family inhibitor comprises ABT-199.

23. The method of claim 22, wherein the BCL-2 family inhibitor is ABT-199.

24. The method of any one of claims 1-21, wherein the BCL-2 family inhibitor comprises ABT-263.

25. The method of claim 24, wherein the BCL-2 family inhibitor is ABT-263.

26. The method of any one of claims 1 to 25, wherein the therapeutically effective amount of the BCL-2 family inhibitor ranges from about 1 to 500 mg / m per day. 2 .

27. The method of any one of claims 1 to 25, wherein the therapeutically effective amount of the BCL-2 family inhibitor ranges from about 10 to 300 mg / m per day. 2 .

28. The method of any one of claims 1 to 27, wherein the therapeutically effective amount of the BCL-2 family inhibitor is less than 1%, eg, less than 10% or less than 25% or less than 50% of the clinically established therapeutic amount.

29. The method of any one of claims 1 to 28, wherein the BCL-2 family inhibitor and the PRMT5 inhibitor are administered sequentially.

30. The method of any one of claims 1 to 28, wherein the BCL-2 family inhibitor and the PRMT5 inhibitor are administered simultaneously.

31. The method of any one of claims 1 to 30, wherein the individual has previously received or completed first-line chemotherapy.

32. The method of claim 31, 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.

33. A method of treating cancer in an individual, the method comprising administering to the individual: Therapeutically effective amount of ABT-199 (Venetoclax): and a therapeutically effective amount of a PRMT5 inhibitor of the formula:

34. A method of treating cancer in an individual, the method comprising administering to the individual: Therapeutically effective amount of ABT-263 (Navitoclax): and A therapeutically effective amount of a PRMT5 inhibitor of the formula:

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