Combination Therapy Using PRMT5 Inhibitors for the Treatment of Cancer

Abstract

The present disclosure relates to methods of treating cancer. The present disclosure further relates to treating cancer in a subject with compounds that are PRMT5 inhibitors, particularly in combination with CDK4 / 6 inhibitors.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 172,643, filed April 8, 2021, and U.S. Provisional Patent Application No. 63 / 253,029, filed October 6, 2021, the disclosures of each of which are incorporated by reference in their entireties herein.

[0002] The present disclosure relates to methods of treating cancer. The present disclosure further relates to treating cancer in a subject with compounds that are protein arginine N-methyltransferase 5 (PRMT5) inhibitors, particularly in combination with cyclin-dependent kinase 4 and 6 (CDK4 / 6) inhibitors. [Background technology]

[0003] 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 functional group of protein L-arginine residues (omega-monomethylation) and the transfer of a second methyl group to the other omega-nitrogen, resulting in symmetric dimethylarginine (sDMA). PRMT5 forms a complex with methylosomal protein 50 (MEP50), which is required for substrate recognition and orientation, and is also required for PRMT5-catalyzed histone 2A and histone 4 methyltransferase activity (see, e.g., Ho et al. (2013) PLoS ONE 8(2):e57008).

[0004] Homozygous deletions of p16 / CDKN2a are common in cancer, and these mutations are generally accompanied by co-deletion of adjacent genes, including the gene encoding methylthioadenosine phosphorylase (MTAP). It is estimated that approximately 15% of all human cancers have 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 levels of the MTAP substrate methylthioadenosine (MTA), a potent PRMT5 inhibitor. Inhibiting PRMT5 activity reduces methylation activity and increases the sensitivity of cell proliferation to PRMT5 depletion or loss of activity. Thus, loss of MTAP activity reduces the methylation activity of PRMT5, rendering cells selectively dependent on PRMT5 activity.

[0006] Despite the importance of PRMT5 to cell viability and the prevalence in cancer, effective therapies that inhibit PRMT5 have been elusive, and there remains a need to develop new PRMT5 inhibitor therapies to treat a wide range of cancers. Summary of the Invention

[0007] One aspect of the present disclosure provides a method for treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of a CDK4 / 6 inhibitor and a therapeutically effective amount of a PRMT5 inhibitor.

[0008] Also provided herein are methods for treating cancer in a subject in need of such treatment. Such methods include determining that the cancer is associated with MTAP homozygous deletion (e.g., is an MTAP-associated cancer). These methods optionally further include determining that the cancer is associated with CDKN2A homozygous deletion. Such methods further include administering to the subject a therapeutically effective amount of a CDK4 / 6 inhibitor and a therapeutically effective amount of a PRMT5 inhibitor.

[0009] 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, with the understanding that the claims are defined by the recitations therein, and not by the specific discussion of the features and advantages described herein.

[0010] 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. These 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 description of the drawings]

[0011] [Figure 1] The results of the method of Example 1 are described in the KRASG12C and CDKN2A / MTAP DEL lung tumor xenograft LU99 model grown in immunodeficient mice. The PRMT5 inhibitor used in this method was MRTX9768 administered at 100 mg / kg twice daily (BID) and the CDK4 / 6 inhibitor was palbociclib administered at 130 mg / kg once daily (QD). Mean tumor volume ± standard error is plotted as the mean value for the indicated study days. [Diagram 2] The results of the method of Example 2 are described in the KRASG12C and CDKN2A / MTAP DEL lung tumor xenograft LU99 model grown in immunodeficient mice. The PRMT5 inhibitor used in this method was MRTX7477 administered BID at 200 mg / kg, and the CDK4 / 6 inhibitor was palbociclib administered QD at 130 mg / kg. Mean tumor volume ± standard error is plotted as the mean value for the indicated study days. [Diagram 3] The results of the method of Example 3 are described in the HCC4006 lung tumor xenograft model. The PRMT5 inhibitor used in this method was MRTX1719 administered QD at 100 mg / kg, and the CDK4 / 6 inhibitor was palbociclib administered QD at 130 mg / kg. The mean tumor volume ± standard error is plotted as the mean value for the indicated study days. [Figure 4]The results of the method of Example 4 are described in the SW1573 PRMT5-044 lung tumor xenograft model. The PRMT5 inhibitor used in this method was MRTX1719 administered QD at 50 mg / kg, and the CDK4 / 6 inhibitor was palbociclib administered QD at 130 mg / kg. Mean tumor volume ± standard error is plotted as the mean value for the indicated study days. [Diagram 5] The results of the method of Example 5 are described in the H1650 lung tumor xenograft model. The PRMT5 inhibitor used in this method was MRTX1719 administered QD at 100 mg / kg, and the CDK4 / 6 inhibitor was palbociclib administered QD at 130 mg / kg. The mean tumor volume ± standard error is plotted as the mean value for the indicated study days. [Figure 6] The results of the method of Example 6 are described in the A549 PRMT-034 lung tumor xenograft model. The PRMT5 inhibitor used in this method was MRTX1719 administered QD at 100 mg / kg, and the CDK4 / 6 inhibitor was palbociclib administered QD at 130 mg / kg. The mean tumor volume ± standard error is plotted as the mean value for the indicated study days. [Figure 7] The results of the method of Example 7 are described in the PANC-05-04 pancreatic tumor xenograft model. The PRMT5 inhibitor used in this method was MRTX1719 administered QD at 100 mg / kg, and the CDK4 / 6 inhibitor was palbociclib administered QD at 130 mg / kg. Mean tumor volume ± standard error is plotted as the mean value for the indicated study days. [Figure 8] The results of the method of Example 8 are described in the BXPC-3 pancreatic tumor xenograft model. The PRMT5 inhibitor used in this method was MRTX1719 administered QD at 100 mg / kg, and the CDK4 / 6 inhibitor was palbociclib administered QD at 130 mg / kg. Mean tumor volume ± standard error is plotted as the mean value for the indicated study days. [Figure 9]The results of the method of Example 9 are described in the MKN45 gastric tumor xenograft model. The PRMT5 inhibitor used in this method was MRTX1719 administered QD at 100 mg / kg, and the CDK4 / 6 inhibitor was palbociclib administered QD at 130 mg / kg. The mean tumor volume ± standard error is plotted as the mean value for the indicated study days. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] 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 can, of course, vary. It is also to be understood 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.

[0013] In view of the present disclosure, the methods and compositions described herein can be adapted by those skilled in the art to meet the desired needs. The present disclosure provides improvements in treating cancer in a subject. As used herein, the terms "subject" or "patient" are used interchangeably and refer to any animal, including mammals, most preferably humans.

[0014] The methods provided herein may be used to treat a wide variety of cancers, including tumors such as lung, prostate, breast, brain, skin, cervical, testicular cancer, etc. More specifically, cancers that may be treated by the compositions and methods of the invention include, but are not limited to, astrocytic, breast, cervical, colorectal, endometrial, esophageal, gastric, head and neck, hepatocellular, laryngeal, lung, oral, ovarian, prostate, and thyroid cancers and sarcomas. More specifically, these compounds can be used to treat: Heart: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma, and teratoma; Lung: bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondroitin hamartoma, mesothelioma; Gastrointestinal: Esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach ( carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, vipoma), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma); genitourinary tract: kidney (adenocarcinoma, Wilms' tumor (nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, metastasis) prostate (adenocarcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenoid tumor, lipoma); liver: liver cancer (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; bile duct: gallbladder cancer, ampullary carcinoma, cholangiocarcinoma; 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 (osteochondroma exostosis), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid, and giant cell tumor; nervous system: skull (osteoma, hemangioma, granuloma, xanthomas, osteitis deformans), meninges (meningioma, meningeal sarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germinoma (pinealoma), glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), spinal neurofibroma, meningioma, glioma, sarcoma);Gynecology: Uterus (endometrial cancer), Cervix (cervical cancer, preneoplastic cervical dysplasia), Ovaries (ovarian cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa theca cell tumor, Sertoli-Leydig cell tumor, dysgerminoma, malignant teratoma), Vulva (squamous cell carcinoma, carcinoma in situ, adenocarcinoma, fibrosarcoma, melanoma), Vagina (clear cell carcinoma, squamous cell carcinoma, fetal rhabdomyosarcoma, sarcoma botryoides (embryonal sarcoma) tumors), fallopian tubes (carcinoma); hematological: blood (myeloid leukemia (acute and chronic), acute lymphocytic leukemia, chronic lymphocytic leukemia, myeloproliferative disorders, multiple myeloma, myelodysplastic syndromes), 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; and adrenal glands: neuroblastoma. ;

[0015] In certain embodiments of the methods of the present disclosure, the cancer is an MTAP-associated cancer. For example, in certain embodiments, the cancer is a MTAP-associated cancer. DEL The subject may be identified or diagnosed as having an MTAP-associated cancer, e.g., DEL is determined using a suitable 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.

[0016] In certain embodiments of the disclosed methods, the cancer is a cancer characterized by a homozygous deletion of the cyclin-dependent kinase inhibitor 2A (CDKN2A) gene (CDKN2A DEL ) The target is CDKN2A DEL Alternatively, the subject may be identified or diagnosed as having CDKN2A, where the deletion is determined using a suitable assay or kit. DEL The subject is suspected of having cancer or the subject is DEL Have clinical documentation showing they have cancer.

[0017] In certain embodiments of the methods of the present disclosure, the cancer is glycine-cysteine ​​(KRAS G12CThe subject may further comprise a Kirsten rat sarcoma viral oncogene homolog (KRAS), such as a KRAS gene mutation. G12C The subject may be identified or diagnosed as having cancer, wherein KRAS G12C The mutation is determined using a suitable assay or kit. Alternatively, the subject is G12C The subject is suspected of having cancer or the subject is G12C Have clinical documentation showing they have cancer.

[0018] In some embodiments of any of the methods or uses described herein, the assay uses a sample from the subject (e.g., a biological sample or a biopsy sample (e.g., a paraffin-embedded biopsy sample)) to determine whether the patient has MTAP DEL and / or CDKN2A DEL and / or KRAS G12C Assays are used to determine whether a gene has a specific function. Such assays include, but are not limited to, next-generation sequencing, immunohistochemistry, fluorescence microscopy, break-apart FISFI analysis, Southern blotting, Western blotting, FACS analysis, Northern blotting, 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 using, for example, at least one labeled nucleic acid probe or at least one labeled antibody or antigen-binding fragment thereof.

[0019] In certain embodiments, the cancer in the methods of the present disclosure is selected from lung cancer, pancreatic cancer, head and neck cancer, bladder cancer, esophageal cancer, lymphoma, gastric cancer, skin cancer, breast cancer, brain cancer, liver cancer, and colon cancer.

[0020] In certain embodiments, the cancer in the methods of the present disclosure is selected from lung cancer, pancreatic cancer, head and neck cancer, bladder cancer, esophageal cancer, lymphoma, gastric cancer, skin cancer, breast cancer, and brain cancer.

[0021] 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, head and neck cancer (such as 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).

[0022] In certain embodiments, the cancer in the methods of the present disclosure is selected from mesothelioma, NSCLC (e.g., adenocarcinoma and squamous cell), pancreatic cancer, HNSCC, and bladder cancer.

[0023] In one embodiment of the disclosed method, the cancer is lung cancer. For example, the lung cancer can be NSCLC (e.g., adenocarcinoma and squamous cell) or mesothelioma.

[0024] In one embodiment of the disclosed methods, the cancer is pancreatic cancer. In another embodiment, the cancer is head and neck cancer. In yet another embodiment, the cancer is bladder cancer.

[0025] As provided above, a CDK4 / 6 inhibitor is administered in the method of the present disclosure. As used herein, "CDK4 / 6 inhibitor" refers to a compound that can negatively regulate or inhibit all or part of the enzymatic activity of CDK4 / 6. The CDK4 / 6 inhibitor of the present disclosure interacts with and / or irreversibly binds to CDK4 / 6, resulting in the inhibition of the enzymatic activity of CDK4 / 6, blocking the transition from G1 to S phase of the cell cycle.

[0026] In certain embodiments, the CDK4 / 6 inhibitor also inhibits CDK2, i.e., the CDK4 / 6 inhibitor is a CDK2 / 4 / 6 inhibitor.

[0027] In certain embodiments, the CDK4 / 6 inhibitor is selected from palbociclib (sold as Ibrance®, Pfizer Inc., New York, New York), abemaciclib (sold as Verzenio®, Eli Lilly and Company, Indianapolis, Indiana), ribociclib (sold as Kisqali®, Novartis Pharmaceuticals, Basel, Switzerland), PF-06873600 (CAS number 2185857-97-8, available from Pfizer Inc., New York, New York), and combinations thereof. In certain embodiments, the CDK4 / 6 inhibitor is trilaciclib (sold as Cosela™, G1 Therapeutics, Inc., Durham, North Carolina).

[0028] In one embodiment of the methods of the disclosure, the CDK4 / 6 inhibitor is palbociclib.

[0029] As provided above, PRMT5 inhibitors are also administered in the methods of the present disclosure.As used herein, "PRMT5 inhibitors" refers to the compounds of the present disclosure described herein.These compounds can negatively regulate or inhibit all or part of the enzyme activity of PRMT5, particularly in the presence of bound MTA in vitro or in vivo, or in cells expressing elevated levels of MTA.In certain embodiments, the PRMT5 inhibitor is an MTA-cooperative PRMT5 inhibitor.

[0030] In certain embodiments, the PRMT5 inhibitor of the present disclosure is any one of the PRMT5 inhibitors disclosed in International Patent Publication No. 2021 / 050915A1, published March 18, 2021, which is incorporated by reference in its entirety.

[0031] In certain other embodiments, the PRMT5 inhibitor of the present disclosure is any one of the PRMT5 inhibitors disclosed in U.S. Provisional Patent Application No. 63 / 200,521, published March 11, 2021, which is incorporated by reference in its entirety.

[0032] For example, the PRMT5 inhibitor in the disclosed methods described herein is a compound of formula IIA, IIB, or IIC: [ka] During the ceremony, A is CR 9 or N, D is (C(R 9 )2) 1-2 -NH2, [ka] or D is [ka] where the methylene is bonded to E and where E is C; E is C, CR 9 , or N, each L is independently a bond or C1-C3 alkylene; W is CR 9 or N, 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)-; R 2 each independently represents hydroxy, halogen, cyano, cyanomethyl, -(NR 4)2, hydroxyalkyl, alkoxy, -SO2C1-C3 alkyl, -X-arC1-C3 alkyl, 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, where heterocyclyl, cycloalkyl, aryl, and heteroaryl are each independently selected from one or more of R 5 and optionally substituted with R 4 are each independently hydrogen or C1-C3 alkyl; R 5 are each independently cyano, oxo, halogen, C1-C3 alkyl, hydroxyalkyl, hydroxy, alkoxy, alkoxy-C1-C3 alkyl, -X-haloalkyl, -Z-cycloalkyl, -X-arC1-C3 alkyl, -X-arC1-C3 alkyl 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; R 9 is each independently H or C1-C3 alkyl, halogen, or haloalkyl, or a pharma- ceutically acceptable salt thereof (embodiment 1).

[0033] Embodiment 2 provides the PRMT5 inhibitor in the methods of the disclosure as a compound of formula IIA. [ka]

[0034] Embodiment 3 provides the PRMT5 inhibitor in the methods of the disclosure as a compound of formula IIB. [ka]

[0035] Embodiment 4 provides the PRMT5 inhibitor in the methods of the disclosure as a compound of formula IIC. [ka]

[0036] In the fifth embodiment, W is CR 9 The method according to any one of embodiments 1 to 4 is provided, wherein

[0037] In embodiment 6, A is CR 9 The method according to any one of embodiments 1 to 4 is provided, wherein

[0038] Embodiment 7 provides a method according to any one of embodiments 1 to 4, wherein E is N.

[0039] In the eighth embodiment, W is CR 9 and A is CR 9 and E is N.

[0040] Embodiment 9 is a compound comprising R 2is selected from benzothiophene, naphthalene, quinoline, chroman, isochroman, dihydrobenzodioxin, indorazine, tetrahydroindolazine, dihydroisobenzofuran, benzene, isoquinolinone, benzodioxone, thienopyridine, tetrahydroindolone, indolizine, dihydroindolizinone, imadazopyridinone, thienopyrimidine, thiophene, pyrrolopyrimidinone, thiazolopyridinone, dihydropyrrolidine, isoindalone, and tetrahydroisoquinoline.

[0041] Embodiment 10 is a method for producing a compound comprising the steps of: 5 is each 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, -XL-cycloalkyl optionally substituted with C1-C3 alkyl or oxo.

[0042] Embodiment 11 is a compound comprising 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.

[0043] Embodiment 12 provides the method of any one of embodiments 1-11, wherein one of L, X, and Z is a bond.

[0044] Embodiment 13 provides the method of embodiment 12, wherein L, X, and Z are all bonds.

[0045] One aspect of the disclosure is that the PRMT5 inhibitor is a compound of formula (IIIC): [ka] During the ceremony, 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 independently C(H), C(R 5 ), and N, with the proviso that only one or two of G, Q, J, and U can be N; R 5 are each 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 and In the formula, R 9 are each independently H or C1-C3 alkyl; R 15 is hydrogen or methyl, and R 16 is C1-C3 alkyl; R 7 is a compound, or a pharma- ceutically acceptable salt thereof, wherein:

[0046] Embodiment 15 provides the method of embodiment 14, wherein A is CH.

[0047] Embodiment 16 provides the method of embodiment 14 or 15, wherein W is N.

[0048] Embodiment 17 provides the method of embodiment 14 or 15, wherein W is CH.

[0049] Embodiment 18 provides the method of any one of embodiments 14-17, wherein D is -CH2-NH2.

[0050] Embodiment 19 provides a method of the disclosure, wherein the PRMT5 inhibitor is a compound according to embodiment 14 of the following formula: [ka]

[0051] Embodiment 20 is a method for producing a compound according to the present invention 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

[0052] Embodiment 21 is a compound comprising 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

[0053] Embodiment 22 is a compound comprising 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.

[0054] Embodiment 23 is a compound comprising 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

[0055] Embodiment 24 is a method for producing a compound according to the present invention 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

[0056] Embodiment 25 is a method for producing a compound according to the present invention 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.

[0057] Embodiment 26 provides the method of any of embodiments 23-25, wherein G, Q, J, and U are each independently C(H).

[0058] Embodiment 27 is an embodiment in which G, Q, J, and U are independently C(H) and C(R 5 The method according to any one of embodiments 23 to 25, wherein the method is selected from the group consisting of:

[0059] 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.

[0060] Embodiment 29 is R 6 is hydrogen, 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), C(R 5 ), and N; R 5 is each 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.

[0061] Embodiment 30 provides the method of embodiment 29, wherein one or two of G, Q, J, and U are N.

[0062] Embodiment 31 is R 6 is hydrogen, 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 ) and R 5is each 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.

[0063] 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), e.g., only one of G, Q, J, and U is C(R 5 32. The method of claim 31, wherein

[0064] 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).

[0065] Embodiment 34 is an embodiment in which three of G, Q, J, and U are C(R 5 ) and the remaining G, Q, J, and U are C(H).

[0066] Embodiment 35 is an embodiment in which G, Q, J, and U, together with the thiophene to which they are attached, represent: [ka] 20. The method according to any one of embodiments 14 to 19, wherein

[0067] Embodiment 36 provides the method of embodiment 35, wherein G, Q, J, and U, together with the thiophene to which they are attached, form a benzo[b]thiophene.

[0068] Embodiment 37 is directed to R 5The method of any one of embodiments 14-36, wherein, if present, 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.

[0069] Embodiment 38 is directed to R 5 The method of any one of embodiments 14-36, wherein, if present, is hydroxy, halogen, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl.

[0070] Embodiment 39 is a method for producing a compound according to the present invention 5 The method of any one of embodiments 14-36, wherein, if present, is hydroxy, chloro, fluoro, methyl, ethyl, methoxy, ethoxy, 2,2-difluoroethoxy, oxetanyl, tetrahydrofuranyl, (methoxy)methyl, (ethoxy)methyl, (methoxy)ethyl, or (ethoxy)ethyl.

[0071] Embodiment 40 is a method for producing a compound according to the present invention 7 The method of any one of embodiments 14-39, wherein is methyl.

[0072] Embodiment 41 is a compound comprising R 7 The method of any one of embodiments 14-39, wherein is ethyl.

[0073] Embodiment 42 is R 7 The method of any one of embodiments 14-39, wherein is propyl (e.g., isopropyl).

[0074] Embodiment 43 is a compound comprising R 7 The method of any one of embodiments 14-39, wherein is difluoromethyl or trifluoromethyl.

[0075] Embodiment 44 is wherein the PRMT5 inhibitor is of the formula: [ka] During the ceremony, G, Q, J, and U together with the thiophene to which they are attached [ka] Forming In the formula, R 5 are each independently hydroxy, halogen, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl; 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 15. The method of embodiment 14, wherein

[0076] Embodiment 45 is wherein the PRMT5 inhibitor is of the formula: [ka] During the ceremony, G, Q, J, and U together with the thiophene to which they are attached [ka] Forming In the formula, R 5are each independently hydroxy, halogen, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl; 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 15. The method of embodiment 14, wherein

[0077] Embodiment 46 is wherein the PRMT5 inhibitor is of the formula: [ka] During the ceremony, G, Q, J, and U together with the thiophene to which they are attached [ka] Forming In the formula, R 5 is each independently hydroxy, halogen, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl.

[0078] Embodiment 47 is a compound of formula (IIIB) [ka] During the ceremony, 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 atom to which they are attached to form a phenyl, R 53 is hydrogen, fluoro, chloro, 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 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 pharma- ceutically acceptable salt thereof.

[0079] Embodiment 48 is A is -CH or -CCH3; D is -CH2-NH2, W is -CH, -CCH3, or N; R51 , 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; R 7 is C1-C2 alkyl or C1-C2 haloalkyl.

[0080] Embodiment 49 is A and W are -CH; D is -CH2-NH2, R 51 , R 52 , and R 53 are each independently selected from hydrogen, fluoro, chloro, and methyl; R 54 is hydrogen, L 5 is -O-, R 6 is hydrogen, R 7 is methyl.

[0081] 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, R 7 is methyl.

[0082] Embodiment 51 provides the method of embodiment 47, wherein A is CH.

[0083] Embodiment 52 provides the method of embodiment 47 or 48, wherein W is N.

[0084] Embodiment 53 provides the method of embodiment 47 or 48, wherein W is CH.

[0085] Embodiment 54 provides the method of any of embodiments 47-50, wherein D is -CH2-NH2.

[0086] Embodiment 55 is directed to R 54 The method of any one of embodiments 47-51, wherein is hydrogen or methyl.

[0087] Embodiment 56 is directed to R 54 The method of any one of embodiments 47-51, wherein is hydrogen.

[0088] Embodiment 57 is directed to R 54 The method of any one of embodiments 47-51, wherein is methyl.

[0089] Embodiment 58 is a method for the preparation of a PRMT5 inhibitor having the formula: [ka] ,for example, [ka] etc.,

[0090] In embodiment 59, L 5 The method of any one of embodiments 47-55, wherein is -CH2-.

[0091] In embodiment 60, L 5The method of any one of embodiments 47-55, wherein is -O-.

[0092] Embodiment 61 is a compound comprising 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 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.

[0093] Embodiment 62 is a compound comprising 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.

[0094] Embodiment 63 is a compound comprising R 6 58. The method of any one of embodiments 47-57, wherein is hydrogen, chloro, fluoro, methyl, ethyl, methoxy, or ethoxy.

[0095] Embodiment 64 is directed to 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 6is 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 65 is directed to R 6 is halogen, C1-C6 alkyl, or C1-C6 alkoxy, for example, R 6 is halogen, C1-C3 alkyl, or C1-C3 alkoxy.

[0097] Embodiment 66 is directed to R 6 58. The method of any one of embodiments 47-57, wherein is chloro, fluoro, methyl, ethyl, methoxy, or ethoxy.

[0098] Embodiment 67 is directed to R 7 The method of any one of embodiments 47-63, wherein is methyl.

[0099] Embodiment 68 is directed to R 7 The method of any one of embodiments 47-63, wherein is ethyl.

[0100] Embodiment 69 is a method for producing 7 The method of any one of embodiments 47-63, wherein is propyl (e.g., isopropyl).

[0101] Embodiment 70 is a method for producing a compound according to the present invention 7 The method of any one of embodiments 47-63, wherein is difluoromethyl or trifluoromethyl.

[0102] Embodiment 71 is R 53 is hydrogen or methoxy, or R 53 The method of any one of embodiments 47-67, wherein is hydrogen.

[0103] Embodiment 72 is a method for the preparation of a PRMT5 inhibitor having the formula: [ka] 48. The method of embodiment 47, wherein

[0104] Embodiment 73 is R 52 is fluoro and R 51 The method of any one of embodiments 47-69, wherein is hydrogen, fluoro, chloro, or methyl.

[0105] Embodiment 74 is R 52 is fluoro and R 51 The method of any one of embodiments 47-69, wherein is chloro.

[0106] Embodiment 75 is a method for producing 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.

[0107] Embodiment 76 is a method for producing 51 and R 52 together with the atom to which they are attached form hydrofuranyl (e.g. [ka] 70. The method of any one of embodiments 47 to 69, further comprising forming a .

[0108] Embodiment 77 is a method for treating a PRMT5 inhibitor comprising administering to a patient [ka] The method of any one of embodiments 47 to 76 is provided, wherein

[0109] Embodiment 78 is a method for treating a PRMT5 inhibitor comprising administering to a patient [ka] The method of any one of embodiments 47 to 77 is provided, wherein

[0110] One aspect of the disclosure is that the PRMT5 inhibitor is a compound of formula (IIIA): [ka] During the ceremony, 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 but, [ka] and In the formula, R 56 is hydrogen, fluoro, chloro, or methyl; G, Q, J, and U are independently C(H), C(R 5 ), and N, with the proviso that only one or two of G, Q, J, and U can be N; R 5are each 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, 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 a compound, or a pharma- ceutically acceptable salt thereof, wherein:

[0111] One aspect of the disclosure is that the PRMT5 inhibitor is a compound of formula (IIIA): [ka] During the ceremony, 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 but, [ka] and In the formula, R 56is hydrogen, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, or C1-C6 haloalkoxy; 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 a compound, or a pharma- ceutically acceptable salt thereof, wherein:

[0112] Embodiment 81 provides the method of embodiment 79 or 80, wherein A is CH.

[0113] Embodiment 82 provides the method of embodiment 79 or 80, wherein W is N.

[0114] Embodiment 83 provides the method of embodiment 79 or 80, wherein W is CH.

[0115] Embodiment 84 provides the method of embodiment 79 or 80, wherein D is -CH2-NH2.

[0116] Embodiment 85 is a compound having the formula: [ka] The method of embodiment 79 or 80 is provided, wherein

[0117] Embodiment 86 is a method for producing a compound according to the present invention 2 but, [ka] The method according to any one of embodiments 79 and 81 to 85, wherein

[0118] Embodiment 87 is an embodiment in which G, Q, J, and U are independently C(H) and C(R 5 87. The method of embodiment 86, wherein the method is selected from the group consisting of

[0119] Embodiment 88 provides the method of embodiment 86, wherein G, Q, J, and U are independently C(H).

[0120] 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), e.g., only one of G, Q, J, and U is C(R 5 87. The method of embodiment 86, wherein

[0121] Embodiment 90 is an embodiment in which U is N and G, Q, and J are independently C(H) and C(R 5 87. The method of embodiment 86, wherein the method is selected from the group consisting of

[0122] Embodiment 91 is an embodiment in which G is N and Q, J, and U are independently C(H) and C(R 5 87. The method of embodiment 86, wherein the method is selected from the group consisting of

[0123] Embodiment 92 is R 5 The method of any one of embodiments 79 or 81-91, wherein, if present, 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.

[0124] Embodiment 93 is R 5The method of any one of embodiments 79 or 81-91, wherein, if present, is hydroxy, halogen, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C3-C6 heterocycloalkyl, or C1-C3 alkoxyC1-C3 alkyl.

[0125] Embodiment 94 is a method for producing 5 The method of any one of embodiments 79 or 81-91, wherein, if present, is hydroxy, chloro, fluoro, methyl, ethyl, methoxy, ethoxy, 2,2-difluoroethoxy, oxetanyl, tetrahydrofuranyl, (methoxy)methyl, (ethoxy)methyl, (methoxy)ethyl, or (ethoxy)ethyl.

[0126] Embodiment 95 is a method for producing 5 When present, it is halogen, C1-C6 alkyl, or C1-C6 alkoxy, e.g., R 6 The method of any one of embodiments 79 or 81-91, wherein is halogen, C1-C3 alkyl, or C1-C3 alkoxy.

[0127] Embodiment 96 is a method for producing 5 The method of any one of embodiments 79 or 81-91, wherein, if present, is chloro, fluoro, methyl, ethyl, methoxy, or ethoxy.

[0128] Embodiment 97 is R 56 The method of any one of embodiments 79 or 81-91, wherein is fluoro, chloro, or methyl.

[0129] Embodiment 98 is R 2 but, [ka] The method according to any one of embodiments 80 to 85, wherein

[0130] Embodiment 99 is R 56 The method of any one of embodiments 80-85 or 98, wherein is hydrogen, fluoro, chloro, or methyl.

[0131] 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 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.

[0132] Embodiment 101 is R 6 is hydrogen, halogen, C1-C6 alkyl, or C1-C6 alkoxy, for example, R 6 The method of any one of embodiments 79-99, wherein is hydrogen, halogen, C1-C3 alkyl, or C1-C3 alkoxy.

[0133] Embodiment 102 is R 6 99. The method of any one of embodiments 79-99, wherein is hydrogen, chloro, fluoro, methyl, ethyl, methoxy, or ethoxy.

[0134] 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)R16 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.

[0135] Embodiment 104 is a method for producing a 6 is halogen, C1-C6 alkyl, or C1-C6 alkoxy, for example, R 6 The method of any one of embodiments 79-99, wherein is halogen, C1-C3 alkyl, or C1-C3 alkoxy.

[0136] Embodiment 105 is a 6 The method of any one of embodiments 79-99, wherein is chloro, fluoro, methyl, ethyl, methoxy, or ethoxy.

[0137] Embodiment 106 is a method for producing a 7 The method of any one of embodiments 79-105, wherein is methyl.

[0138] Embodiment 107 is R 7 The method of any one of embodiments 79-105, wherein is ethyl.

[0139] Embodiment 108 is a method for producing a 7 The method of any one of embodiments 79-105, wherein is propyl (e.g., isopropyl).

[0140] Embodiment 109 is R 7 The method of any one of embodiments 79-105, wherein is difluoromethyl or trifluoromethyl.

[0141] In certain embodiments of the disclosed methods described herein, the PRMT5 inhibitor is [ka] It is.

[0142] In certain embodiments of the disclosed methods described herein, the PRMT5 inhibitor is [ka] It is.

[0143] In certain embodiments of the disclosed methods described herein, the PRMT5 inhibitor is [ka] It is.

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

[0145] The PRMT5 inhibitors and / or CDK4 / 6 inhibitors of the present disclosure may be provided as pharmaceutical compositions comprising a therapeutically effective amount of such inhibitors and pharma- ceutically acceptable carriers, excipients, and / or diluents. The PRMT5 inhibitors and / or CDK4 / 6 inhibitors of the present disclosure may be formulated by any method known in the art and prepared for administration by any route, including, but not limited to, parenteral, oral, sublingual, transdermal, topical, intranasal, intratracheal, or intrarectal. In certain embodiments, the PRMT5 inhibitors and / or CDK4 / 6 inhibitors of the present disclosure are administered intravenously in a hospital setting. In certain other embodiments, administration may be preferably by oral route.

[0146] The characteristics of the carrier will depend on the route of administration. As used herein, the term "pharmaceutical acceptable" refers to a non-toxic material 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. Thus, the pharmaceutical composition of the present disclosure may contain, in addition to the inhibitor, diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art. The preparation of pharmaceutical acceptable formulations is described, for example, in Remington's Pharmaceutical Sciences, 18 th Edition, ed. A. Gennaro, Mack Publishing Co., Easton, Pa., 1990.

[0147] The PRMT5 inhibitors and CDK4 / 6 inhibitors of the present disclosure are administered in a therapeutically effective amount. As used herein, the phrase "therapeutically effective amount" or "effective amount" refers to an amount of active agent that induces a biological or pharmaceutical response sought in a tissue, system, subject, or human by a researcher, physician, or other clinician. Generally, a therapeutically effective amount is sufficient to deliver a biological or pharmaceutical response to a subject without causing significant toxic effects. Doses of active agents can range from about 0.01 to 300 mg / kg per day, e.g., 0.1 to 100 mg / kg per day, more commonly 0.5 to about 25 mg per kg of recipient body weight per day. Typical local dosages will be in the range of 0.01 to 3% weight / weight in a suitable carrier.

[0148] In certain embodiments of the disclosed methods, the therapeutically effective amount of the PRMT5 inhibitor is in the range of about 0.01-300 mg / kg per day. For example, in certain embodiments, the therapeutically effective amount of the PRMT5 inhibitor is in the range of about 0.1-100 mg / kg per day, or 25-100 mg / kg per day, or 50-100 mg / kg per day.

[0149] In certain embodiments, 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 (e.g., the amount required when the PRMT5 inhibitor is administered alone).

[0150] In certain embodiments of the disclosed methods, the therapeutically effective amount of the CDK4 / 6 inhibitor ranges from about 0.01 to 300 mg / kg per day. For example, in certain embodiments, the therapeutically effective amount of the CDK4 / 6 inhibitor ranges from about 0.1 to 100 mg / kg per day, or 0.1 to 50 mg / kg per day, or 10 to 100 mg / kg per day, or 10 to 50 mg / kg per day.

[0151] In certain embodiments, the therapeutically effective amount of the CDK4 / 6 inhibitor is less than 1%, e.g., less than 10%, or less than 25%, or less than 50% of the clinically established therapeutic amount (e.g., the amount required when the CDK4 / 6 inhibitor is administered alone).

[0152] Combination therapy, in defining the use of the PRMT5 inhibitor and CDK4 / 6 inhibitor of the present disclosure, is intended to embrace sequential administration of each agent in a regimen that results in the beneficial effect of the drug combination (e.g., the PRMT5 inhibitor and CDK4 / 6 inhibitor of the present disclosure can be formulated as separate compositions given sequentially), as well as embrace the simultaneous administration of these agents substantially simultaneously, for example, in a single dosage form with a fixed ratio of these active agents, or in multiple or separate dosage forms for each agent. The present disclosure is not limited to the order of administration: the PRMT5 inhibitor of the present disclosure can be administered either before or after (i.e., sequentially), or simultaneously (i.e., simultaneously) the administration of the CDK4 / 6 inhibitor of the present disclosure.

[0153] The methods of the present disclosure are useful as a first line treatment. Thus, in certain embodiments of the methods of the present disclosure, the subject has not previously received another first line treatment.

[0154] The method of the present disclosure is also useful as first-line maintenance or second-line treatment. Thus, in certain embodiments of the method of the present disclosure, the subject has previously completed another first-line therapy. For example, in certain embodiments, the method of the present disclosure can provide delay in cancer progression and recurrence in subjects who have previously completed another first-line chemotherapy. For example, in certain embodiments, the subject has previously completed platinum-based and / or taxane-based chemotherapy (e.g., carboplatin, cisplatin, oxaliplatin, paclitaxel, docetaxel, etc.). In certain embodiments of the method of the present disclosure, the subject has previously completed another first-line chemotherapy and is partially responsive to such chemotherapy.

[0155] definition For the sake of brevity, chemical moieties are defined and referred to throughout primarily as monovalent chemical moieties (e.g., alkyl, aryl, etc.). Nevertheless, such terms may also be used to convey corresponding multivalent moieties under appropriate structural circumstances that are apparent to one of skill in the art. For example, while an "alkyl" moiety generally refers to a monovalent radical (e.g., CH3-CH2-), under certain circumstances, a divalent linking moiety may be an "alkyl", in which case one of skill in the art would understand that the alkyl is the divalent radical equivalent to the term "alkylene" (e.g., -CH2-CH2-). (Similarly, in situations where a divalent moiety is required and described as "aryl", one of skill in the art would understand that the term "aryl" refers to the corresponding divalent moiety, arylene). It is understood that all atoms have their normal valence 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).

[0156] The term "amino" refers to --NH.sub.2.

[0157] The term "acetyl" refers to -C(O)CH3.

[0158] As used herein, the term "acyl" refers to an alkylcarbonyl or arylcarbonyl substituent, where the alkyl and aryl moieties are as defined herein.

[0159] The term "alkyl" as used herein 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, C57 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.

[0160] The term "alkenyl" as used herein means an unsaturated straight or branched chain aliphatic group containing one or more carbon-carbon double bonds 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, C53, C5 10 , C 11 , and C 12 Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, and hexenyl.

[0161] The term "alkynyl" as used herein refers to an unsaturated straight or branched chain aliphatic group containing one or more carbon-carbon triple bonds having 2 to 12 carbon atoms. Thus, "alkynyl" 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, C53, C 10 , C 11 , and C 12 Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, and hexynyl.

[0162] An "alkylene", "alkenylene", or "alkynylene" group is an alkyl, alkenyl, or alkynyl group, as defined hereinabove, that is located 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.

[0163] The term "alkoxy" refers to -OC1-C6 alkyl.

[0164] The term "cycloalkyl" as used herein refers to saturated and partially unsaturated cyclic hydrocarbon groups having 3 to 12 carbons. Thus, "cycloalkyl" includes C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C210, C220, C23, C24, C25, 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.

[0165] The term "heteroalkyl" is as defined herein above, and one or more of the carbon atoms in the chain are independently O, S, or NR x and R x is hydrogen or C1-C3 alkyl. Examples of heteroalkyl groups include methoxymethyl, methoxyethyl, and methoxypropyl.

[0166] An "aryl" group is a C6-C aryl group containing one to three aromatic rings. 14 It is an aromatic moiety. Therefore, "aryl" includes C6, C 10 , C 13 , and C 14Exemplary aryl groups include C6-C 10 An aryl group. Particular aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, and fluorenyl. An "aryl" group includes 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, such as indenyl.

[0167] An "aralkyl" or "arylalkyl" group comprises an aryl group covalently bonded to an alkyl group, which in turn is bonded to another group via the alkyl moiety. Exemplary aralkyl groups include, but are not limited to, -(C1-C6)alkyl(C6-C8)alkyl, -(C1 ... 10 ) aryl. For example, arC1-C3 alkyl is an aryl group covalently linked to a C1-C3 alkyl.

[0168] A "heterocyclyl" or "heterocyclic" group is a mono- or bicyclic (fused or spiro) ring structure having three to twelve atoms (3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 atoms), e.g., four to eight atoms, in which one or more ring atoms are independently selected from -C(O)-, N, NR 4 , O, or S, with the remainder of the ring atoms being quaternary 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, dimethyl-morpholinyl, and morpholinyl. Specifically excluded from the scope of this term are compounds having adjacent cyclic O and / or S atoms.

[0169] As used herein, "L-heterocyclyl" refers to a heterocyclyl group covalently linked to another group via an alkylene linker.

[0170] 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 within the cyclic array, 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, 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, oxa Zolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, 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, quinucinyl Examples include lysinyl, 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.

[0171] 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, benzimidazolylethyl, quinazolinylmethyl, quinolinylmethyl, quinolinylethyl, benzofuranylmethyl, indolinylethyl, isoquinolinylmethyl, isoinodylmethyl, cinnolinylmethyl, and benzothiophenylethyl. Compounds having adjacent cyclic O and / or S atoms are specifically excluded from the scope of this term.

[0172] An "arylene," "heteroarylene," or "heterocyclylene" group is a divalent aryl, heteroaryl, or heterocyclyl group, respectively, as defined hereinabove, that is positioned between and serves to connect two other chemical groups.

[0173] As used herein, when a moiety (e.g., cycloalkyl, aryl, heteroaryl, heterocyclyl, urea, etc.) is described as "optionally substituted" without explicitly reciting substituents, it means that the group optionally has 1 to 4, preferably 1 to 3, and more preferably 1 or 2 non-hydrogen substituents.

[0174] The term "halogen" or "halo" as used herein refers to chlorine, bromine, fluorine, or iodine.

[0175] 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.

[0176] The term "hydroxyalkyl" refers to -alkylene-OH. EXAMPLES

[0177] The methods of the present disclosure are further illustrated by the following examples, which are not to be construed as limiting the disclosure in scope or spirit to the specific procedures and compounds described therein.

[0178] Study design: The PRMT5 inhibitors of the present disclosure show selective activity in MTAP-deficient cancers by binding to and inhibiting PRMT5 when bound to the intracellular metabolite MTA.As mentioned above, MTAP is an enzyme in the methionine salvage pathway, and its deletion 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 of the present disclosure represent a novel approach to selectively treat MTAP-deficient cancers.

[0179] It is highly unlikely that a single mutation will cause cancer, and in most cases, it is multiple mutations that cause cancer to develop.The present inventors have found that the use of combination therapy improves the treatment of certain cancers with PRMT5 inhibitors.In particular, the present inventors have surprisingly found that combination therapy of PRMT5 inhibitors and CDK4 / 6 inhibitors provides higher antitumor activity compared to either inhibitor alone.

[0180] Research Procedure: Immunodeficient female nu / nu mice were treated with 5 × 10 6 The tumors were approximately 150–200 mm 3Tumor volumes were measured using calipers until the tumor volume reached 100%. Animals were randomized to receive A) vehicle (0.5% methylcellulose (4000cps) / 0.2% Tween80 in water), B) PRMT5 inhibitor, C) CDK4 / 6 inhibitor, or D) PRMT5 inhibitor and CDK4 / 6 inhibitor, all administered orally (PO) for 21 days. Tumor volumes were measured twice weekly (n=5 / treatment group). Mean tumor volumes and standard error of the mean were calculated and plotted in GraphPad for each study day.

[0181] Example 1 This example was carried out according to the above study procedure. The PRMT5 inhibitor was MRTX9768, administered at 100 mg / kg twice daily (BID). MRTX9768 is 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-3-fluoro-1-naphthonitrile, and is disclosed as Example 16-1 on page 304 of International Patent Publication No. 2021 / 050915A1, published March 18, 2021, which is incorporated by reference in its entirety.

[0182] The CDK4 / 6 inhibitor used in this example was palbociclib administered at 130 mg / kg once daily (QD). Palbociclib is 6-acetyl-8-cyclopentyl-5-methyl-2-[(5-piperazin-1-ylpyridin-2-yl)amino]pyrido[2,3-d]pyrimidin-7-one, sold as Ibrance®, and available from Pfizer Inc., New York, New York.

[0183] The results are presented in Figure 1 and Table 1. The combination of MRTX9768 and MRTX849 inhibited this KRAS G12C and CDKN2A / MTAP DEL It resulted in greater antitumor activity compared to either inhibitor alone in the lung tumor xenograft LU99 model. [Table 1]

[0184] Example 2 This example was carried out according to the study procedure described above. The PRMT5 inhibitor was MRTX7477, administered BID at 200 mg / kg. MRTX7477 is 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-1-naphthonitrile, and is disclosed as Example 4-147 on page 226 of International Patent Publication No. 2021 / 050915A1, published March 18, 2021, which is incorporated by reference in its entirety. The CDK4 / 6 inhibitor used in this example was the same as palbociclib in Example 1, administered QD at 130 mg / kg.

[0185] The results are presented in Figure 2 and Table 2. The combination of MRTX7477 and MRTX849 inhibited this KRAS G12C and CDKN2A / MTAP DEL It resulted in greater antitumor activity compared to either inhibitor alone in the lung tumor xenograft LU99 model. [Table 2]

[0186] Examples 3 to 6 Compounds of the present disclosure were evaluated in several different lung tumor xenograft models: HCC4006, SW1573 PRMT5-044, H1650, and A549 PRMT-034. This example was carried out substantially according to the study procedure described above, except that mice bearing HCC4006 xenograft tumors, SW1573 PRMT5-044 xenograft tumors, H1650 xenograft tumors, or A549 PRMT-034 xenograft tumors were used. The PRMT5 inhibitor was MRTX1719 administered at 50 mg / kg QD or 100 mg / kg 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. 2021 / 050915A1, published March 18, 2021, which is incorporated by reference in its entirety. The CDK4 / 6 inhibitor used in this example was the same as palbociclib in Example 1, administered QD at 130 mg / kg.

[0187] The results for HCC4006 are provided in FIG. [Table 3]

[0188] The results for SW1573 PRMT5-044 are provided in FIG. [Table 4]

[0189] The results for H1650 are provided in FIG. [Table 5]

[0190] The results for A549 PRMT-034 are provided in Figure 6 and Table 6. [Table 6]

[0191] Examples 7 and 8 The compounds of the present disclosure were evaluated in two different pancreatic tumor xenograft models: PANC-05-04 and BXPC-3. This example was carried out substantially according to the study procedure described above, except that mice bearing PANC-05-04 or BXPC-3 xenograft tumors were used. The PRMT5 inhibitor and CDK4 / 6 inhibitor used in this example were the same as those in Example 3. MRTX1719 was administered QD at 100 mg / kg, and palbociclib was administered QD at 130 mg / kg.

[0192] The results for PANC-05-04 are provided in Figure 7 and Table 7. [Table 7]

[0193] The results for BXPC-3 are provided in FIG. [Table 8]

[0194] Example 9 Compounds of the present disclosure were evaluated in the gastric tumor xenograft model MKN45, and the results are provided in Figure 9 and Table 9. This example was carried out substantially according to the study procedures described above, except that mice bearing MKN45 xenograft tumors were administered. The PRMT5 inhibitor and CDK4 / 6 inhibitor used in this example were the same as those in Example 3. MRTX1719 was administered QD at 100 mg / kg, and palbociclib was administered QD at 130 mg / kg. [Table 9]

[0195] It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes will be suggested to those skilled in the art in light thereof and are incorporated within the spirit and scope of this application and the appended claims. All publications, patents, and patent applications cited herein are incorporated by reference herein for all purposes.

Claims

1. 1. A pharmaceutical composition for treating cancer in a subject, comprising: The pharmaceutical composition comprises a therapeutically effective amount of a cyclin-dependent kinase 4 and 6 (CDK4 / 6) inhibitor and a therapeutically effective amount of a protein arginine N-methyltransferase 5 (PRMT5) inhibitor.

2. The pharmaceutical composition of claim 1, wherein the cancer comprises a homozygous deletion of the methylthioadenosine phosphorylase (MTAP) gene.

3. 2. The pharmaceutical composition of claim 1, wherein the cancer comprises a homozygous deletion of the cyclin-dependent kinase inhibitor 2A (CDKN2A) gene.

4. The cancer is characterized in that the cancer is caused by Kirsten rat sarcoma viral oncogene homolog glycine-cysteine ​​(KRAS) Ｇ１２Ｃ 4. The pharmaceutical composition of claim 2 or 3, further comprising a genetic mutation.

5. 2. The pharmaceutical composition of claim 1, wherein the cancer is lung cancer, pancreatic cancer, head and neck cancer, bladder cancer, esophageal cancer, lymphoma, gastric cancer, skin cancer, breast cancer, brain cancer, liver cancer, and colon cancer.

6. 2. The pharmaceutical composition of claim 1, wherein the cancer is lung cancer (e.g., mesothelioma or non-small cell lung cancer (NSCLC) including adenocarcinoma and squamous cell), pancreatic cancer, head and neck cancer, 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).

7. The pharmaceutical composition of claim 1 , wherein the CDK4 / 6 inhibitor is CDK2 / 4 / 6.

8. 2. The pharmaceutical composition of claim 1, wherein the CDK4 / 6 inhibitor is selected from the group consisting of palbociclib, abemaciclib, ribociclib, PF-06873600, and combinations thereof.

9. 2. The pharmaceutical composition of claim 1, wherein the CDK4 / 6 inhibitor is palbociclib.

10. The pharmaceutical composition of claim 1 , wherein the PRMT5 inhibitor is a methylthioadenosine (MTA)-cooperative PRMT5 inhibitor.

11. The PRMT5 inhibitor is a compound of formula IIA, formula IIB, or formula IIC, 【Chemistry 1】 During the ceremony, A is CR ９ or N, Dが、(C(R ９ ) ２ ) １－２ -NH ２ 、 【Chemistry 2】 or D is 【Chemistry 3】 wherein said methylene is attached to E, and wherein E is C; E is C, CR ９ or N, Each L is independently a bond or C １ -C ３ is 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-, or -SO ２ -, -CH(OH)-, or -C(O)-; R ２ each independently represents hydroxy, halogen, cyano, cyanomethyl, -(NR ４ ) ２ , hydroxyalkyl, alkoxy, -SO ２ C １ -C ３ Alkyl, -X-arC １ -C ３ Alkyl, Heteroalkyl, C ２ -C ４ Alkynyl, -X-haloalkyl, -X-C １ -C ５ Alkyl, -Z-C １ -C ５ alkyl, heterocyclyl, -XL-cycloalkyl, -Z-cycloalkyl, -X-aryl, -Z-aryl, or -X-heteroaryl, wherein said heterocyclyl, said cycloalkyl, said aryl, and said heteroaryl are each independently selected from one or more of R ５ and optionally substituted with R ４ are each independently hydrogen or C １ -C ３ is alkyl, R ５ each independently represents cyano, oxo, halogen, C １ -C ３ Alkyl, hydroxyalkyl, hydroxy, alkoxy, alkoxy-C １ -C ３ Alkyl, -X-haloalkyl, -Z-cycloalkyl, -X-arC １ -C ３ -X-arC substituted with alkyl or cyano １ -C ３ Alkyl, C １ -C ３ -XL-cycloalkyl optionally substituted with alkyl or oxo, one or more C １ -C ３ -XL-heteroaryl optionally substituted with alkyl or oxo, one or more C １ -C ３ -XL-heterocyclyl, or -X-aryl, optionally substituted with alkyl or oxo; R ６ But hydrogen, halogen, C １ -C ３ Alkyl, haloalkyl, hydroxy, alkoxy, C １ -C ３ Alkyl-alkoxy, N(R ９ ) ２ , N.R. ９ C(O)R ９ , C(O)R ９ , oxetane, and THF; R ７ is optionally substituted with H or one or more halogens; １ -C ３ is alkyl, R ８ is H or C １ -C ３ is alkyl, and R ９ are each independently H or C １ -C ３ alkyl, halogen, or haloalkyl, or a pharma- ceutically acceptable salt thereof; or The PRMT5 inhibitor is a compound of formula IIIA, 【Chemistry 4】 During the ceremony, A is CR ９ or N, D is -CH ２ -NH ２ , 【Chemistry 5】 and W is CR ９ or N, where R ９ is H or C １ -C ３ is alkyl, R ２ but, 【Chemistry 6】 and In the formula, R ５６ is hydrogen, fluoro, chloro, or methyl; G, Q, J, and U are independently C(H), C(R ５ ), and N, with the proviso that only one or two of G, Q, J, and U can be N; R ５ each independently represents 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 is alkyl, R ６ But hydrogen, halogen, C １ -C ６ Alkyl, Hydroxy, 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 Wherein R １５ is hydrogen or methyl, R １６ C １ -C ３ is alkyl, and R ７ But, C １ -C ３ Alkyl or C １ -C ３ haloalkyl, or a pharma- ceutically acceptable salt thereof; or The PRMT5 inhibitor is a compound of formula IIIB, 【Chemistry 7】 During the ceremony, A is CR ９ or N, D is -CH ２ -NH ２ , 【Chemistry 8】 and W is CR ９ or N, where R ９ is H or C １ -C ３ is alkyl, R ５１ is hydrogen, fluoro, chloro, or methyl, or R ５１ and R ５２ together with the atoms to which they are attached, C ４ -C ６ forming a heterocycloalkyl (e.g., hydrofuranyl); R ５２ is fluoro, chloro, or methyl, or R ５２ and R ５３ together with the atom to which they are attached to form a phenyl, R ５３ is hydrogen, fluoro, chloro, or methyl; R ５４ But hydrogen, halogen, C １ -C ３ Alkyl, or C １ -C ３ is an alkoxy; L ５ is -O- or -CH ２ - and R ６ But hydrogen, halogen, C １ -C ６ Alkyl, Hydroxy, 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 Wherein R １５ is hydrogen or methyl, R １６ C １ -C ３ is alkyl, R ７ But, C １ -C ３ Alkyl or C １ -C ３ haloalkyl, or a pharma- ceutically acceptable salt thereof; or The PRMT5 inhibitor is a compound of formula IIIC, 【Chemistry 9】 During the ceremony, A is CR ９ or N, D is -CH ２ -NH ２ , 【Chemistry 10】 and W is CR ９ or N, where R ９ is H or C １ -C ３ is alkyl, G, Q, J, and U are independently C(H), C(R ５ ), and N, with the proviso that only one or two of G, Q, J, and U can be N; R ５ each independently represents 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 is alkyl, R ６ But hydrogen, halogen, C １ -C ６ Alkyl, C １ -C ６ Haloalkyl, Hydroxy, 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 and In the formula, R ９ are each independently H or C １ -C ３ alkyl, R １５ is hydrogen or methyl, R １６ C １ -C ３ is alkyl, and R ７ But, C １ -C ３ Alkyl or C １ -C ３ haloalkyl, or a pharma- ceutically acceptable salt thereof. The pharmaceutical composition of claim 1.

12. The PRMT5 inhibitor is 【Chemistry 11】 or a pharma- ceutically acceptable salt thereof.

13. The PRMT5 inhibitor is 【Chemistry 12】 or a pharma- ceutically acceptable salt thereof.

14. 2. The pharmaceutical composition of claim 1, wherein the PRMT5 inhibitor is MRTX1719 or a pharma- ceutically acceptable salt thereof and the CDK4 / 6 inhibitor is palbociclib.

15. The method of claim 1 , wherein the subject has previously undergone or completed first-line chemotherapy.

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