How to Treat Cancer

Abstract

The disclosure features useful methods for treating, for example, cancer in a subject in need thereof. In some embodiments, the described methods include administering an agent that reduces the level and / or activity of BRM / BRG1 in combination with immunotherapy.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 227,111, filed July 29, 2021, and U.S. Provisional Application No. 63 / 280,430, filed November 17, 2021, the contents of which are incorporated herein by reference in their entireties. [Background technology]

[0002] The present invention relates to methods of treating cancer with compounds that modulate BRG1 or the BRM-associated factor (BAF) complex.

[0003] Chromatin regulation is essential for gene expression, and ATP-dependent chromatin remodeling is the mechanism by which such gene expression occurs. The human switch / sucrose nonfermenting (SWI / SNF) chromatin remodeling complex, also known as the BAF complex, has two SWI2-like ATPases known as BRG1 (Brahma-related gene-1) and BRM (Brahma). The transcriptional activator BRG1, also known as the ATP-dependent chromatin remodeler SMARCA4, is encoded by the SMARCA4 gene on chromosome 19. BRG1 is overexpressed in some cancer tumors and is required for cancer cell proliferation. BRM, also known as the likely global transcriptional activator SNF2L2 and / or the ATP-dependent chromatin remodeler SMARCA2, is encoded by the SMARCA2 gene on chromosome 9, and has been shown to be essential for tumor cell proliferation in cells characterized by loss-of-function mutations in BRG1. Deactivation of BRG and / or BRM leads to downstream effects in cells, including cell cycle arrest and tumor suppression.

[0004] Immunotherapy, which harnesses the patient's immune system, has been found to be effective in treating many different cancer types and has become an important part of cancer therapy. However, some cancers respond poorly to immunotherapy. Therefore, there is a need to develop approaches to increase the responsiveness of cancers to immunotherapy. Summary of the Invention

[0005] The present invention features useful methods for treating, for example, cancer in a subject in need thereof. In some embodiments, the methods described herein include administration of a BRM and / or BRG-1 inhibitor in combination with immunotherapy for the treatment of cancer.

[0006] In one aspect, the invention features a method of treating cancer in a subject in need thereof, the method including administering to the subject (i) an effective amount of an agent that reduces a level and / or activity of BRM and / or BRG1 in the subject, and (ii) an effective amount of an immunotherapy.

[0007] In some embodiments, the immunotherapy is administered simultaneously with the agent that reduces the level and / or activity of BRM and / or BRG1 in the subject. In some embodiments, the immunotherapy is administered before (e.g., at least 1 hour, at least 12 hours, at least 1 day, at least 1 week, at least 2 weeks, at least 4 weeks) the agent that reduces the level and / or activity of BRM and / or BRG1 in the subject. In some embodiments, the immunotherapy is administered after (e.g., at least 1 hour, at least 12 hours, at least 1 day, at least 1 week, at least 2 weeks, at least 4 weeks) the agent that reduces the level and / or activity of BRM and / or BRG1 in the subject.

[0008] In some embodiments, the cancer has not responded to a previously administered immunotherapy, hi some embodiments, the cancer is resistant (e.g., determined or predicted to be resistant) to immunotherapy.

[0009] In one embodiment of any of the above methods, the immunotherapy is a CTLA-4 inhibitor, a PD-1 inhibitor, a PD-L1 inhibitor, or an adoptive T cell transfer therapy. In some embodiments, the immunotherapy is a PD-1 inhibitor such as a PD-1 antibody, a PD-L1 inhibitor such as a PD-L1 antibody, a CTLA-4 inhibitor such as a CTLA-4 antibody or fusion protein, a CSF-1R inhibitor, an IDO inhibitor, an A1 adenosine inhibitor, an A2A adenosine inhibitor, an A2B adenosine inhibitor, an A3A adenosine inhibitor, an arginase inhibitor, or an HDAC inhibitor. In some embodiments, the immunotherapy is a PD-1 inhibitor (e.g., nivolumab, pembrolizumab, pidilizumab, or BMS 936559). In some embodiments, the immunotherapy is a PD-L1 inhibitor (e.g., atezolizumab or MEDI4736). In some embodiments, the immunotherapy is a CTLA-4 inhibitor (e.g., ipilimumab). In some embodiments, the immunotherapy is a CSF-1R inhibitor (e.g., pexidartinib or AZD6495). In some embodiments, the immunotherapy is an IDO inhibitor (e.g., norharmane, rosmarinic acid, or α-methyl-tryptophan). In some embodiments, the immunotherapy is an A1 adenosine inhibitor (e.g., 8-cyclopentyl-1,3-dimethylxanthine, 8-cyclopentyl-1,3-dipropylxanthine, 8-phenyl-1,3-dipropylxanthine, bamifylline, BG-9719, BG-9928, FK-453, FK-838, rolofylline, or N-0861). In some embodiments, the immunotherapy is an A2A adenosine inhibitor (e.g., ATL-4444, istradefylline, MSX-3, preladenant, SCH-58261, SCH-412,348, SCH-442,416, ST-1535, VER-6623, VER-6947, VER-7835, viadenant, or ZM-241,385). In some embodiments, the immunotherapy is an A2B adenosine inhibitor (e.g., ATL-801, CVT-6883, MRS-1706, MRS-1754, OSIP-339,391, PSB-603, PSB-0788, or PSB-1115).In some embodiments, the immunotherapy is an A3A adenosine inhibitor (e.g., KF-26777, MRS-545, MRS-1191, MRS-1220, MRS-1334, MRS-1523, MRS-3777, MRE-3005-F20, MRE-3008-F20, PSB-11, OT-7999, VUF-5574, or SSR161421). In some embodiments, the immunotherapy is an arginase inhibitor (e.g., an arginase antibody, (2s)-(+)-amino-5-iodoacetamidopentanoic acid, NG-hydroxy-L-arginine, (2S)-(+)-amino-6-iodoacetamidohexanoic acid, or (R)-2-amino-6-borono-2-(2-(piperidin-1-yl)ethyl)hexanoic acid). In some embodiments, the immunotherapy is an HDAC inhibitor (eg, valproic acid, SAHA, or romidepsin).

[0010] In some embodiments, the immunotherapy is a CD-161 (also known as KLRB1 or NKR-P1A) inhibitor (e.g., IMT-009). In some embodiments, the immunotherapy is an NK and T cell modulator (e.g., IMT-073).

[0011] In some embodiments, an effective amount of an agent that reduces the level and / or activity of BRM and / or BRG1 in a subject (e.g., reduces the activity level by at least 5%, at least 10%, at least 20%, at least 50%, at least 70%, at least 90%, at least 95%, at least 99%) is an amount effective to increase the level of activated T cells in the subject (e.g., in the tumor microenvironment).

[0012] In some embodiments, the cancer expresses BRG1 and / or BRM proteins and / or the cells or subject have been identified as expressing BRG1 and / or BRM. In some embodiments, the cancer expresses BRG1 protein and / or the cells or subject have been identified as expressing BRG1. In some embodiments, the cancer expresses BRM protein and / or the cells or subject have been identified as expressing BRM. In some embodiments, the subject or cancer has and / or has been identified as having a loss-of-function mutation in BRG1. In some embodiments, the subject or cancer has and / or has been identified as having a loss-of-function mutation in BRM.

[0013] In some embodiments of any of the above methods, the cancer has or has been determined to have one or more BRG1 mutations (e.g., homozygous mutations). In some embodiments, the one or more BRG1 mutations include a mutation in the ATPase catalytic domain of the protein. In some embodiments, the one or more BRG1 mutations include a deletion at the C-terminus of BRG1.

[0014] In some embodiments of any of the above methods, the cancer is determined to have or not have an epidermal growth factor receptor (EGFR) mutation. In some embodiments of any of the above methods, the cancer is determined to have or not have an anaplastic lymphoma kinase (ALK) driver mutation. In some embodiments of any of the above methods, the cancer is determined to have or to have a KRAS mutation.

[0015] In some embodiments, the cancer has or has been determined to have a mutation in GNAQ. In some embodiments, the cancer has or has been determined to have a mutation in GNA11. In some embodiments, the cancer has or has been determined to have a mutation in PLCB4. In some embodiments, the cancer has or has been determined to have a mutation in CYSLTR2. In some embodiments, the cancer has or has been determined to have a mutation in BAP1. In some embodiments, the cancer has or has been determined to have a mutation in SF3B1. In some embodiments, the cancer has or has been determined to have a mutation in EIF1AX. In some embodiments, the cancer has or has been determined to have a TFE3 rearrangement. In some embodiments, the cancer has or has been determined to have a TFEB rearrangement. In some embodiments, the cancer has or has been determined to have a MITF rearrangement. In some embodiments, the cancer has or has been determined to have an EZH2 mutation. In some embodiments, the cancer has or has been determined to have a SUZ12 mutation. In some embodiments, the cancer has, or has been determined to have, an EED mutation.

[0016] In some embodiments, the cancer is metastatic. For example, the cancer comprises cells that exhibit migratory cell migration and / or invasion, and / or cells that exhibit endothelial recruitment and / or angiogenesis. Metastatic cancer can spread via seeding the surfaces of the peritoneum, pleura, pericardium, or subarachnoid space. Alternatively, metastatic cancer can spread via the lymphatic system or hematogenously. In some embodiments, the cancer is a cell migration cancer (e.g., a non-metastatic cell migration cancer).

[0017] In some embodiments of any of the above methods, the cancer is non-small cell lung cancer, colon cancer, bladder cancer, cancer of unknown primary site, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, esophagogastric cancer, esophageal cancer, pancreatic cancer, hepatobiliary cancer, soft tissue sarcoma, ovarian cancer, head and neck cancer, renal cell carcinoma, bone cancer, non-Hodgkin's lymphoma, small cell lung cancer, prostate cancer, embryonal tumor, germ cell tumor, cervical cancer, thyroid cancer, salivary gland cancer, gastrointestinal neuroendocrine tumor, uterine sarcoma, gastrointestinal stromal tumor, CNS cancer, thymus tumor, adrenocortical carcinoma, appendix cancer, small intestine cancer, penile cancer, bone cancer, or blood cancer.In some embodiments of any of the above methods, the cancer is esophageal cancer.

[0018] In some embodiments of any of the aforementioned methods, the cancer is non-small cell lung cancer, colon cancer, bladder cancer, cancer of unknown primary, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, penile cancer, bone cancer, renal cell carcinoma, prostate cancer, or blood cancer. In some embodiments of any of the aforementioned methods, the cancer is non-small cell lung cancer.

[0019] In some embodiments of any of the aforementioned methods, the cancer is melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or blood cancer.

[0020] In some embodiments, the cancer is melanoma (e.g., uveal melanoma, mucosal melanoma, or cutaneous melanoma). In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is hematological cancer (e.g., multiple myeloma, large cell lymphoma, acute T-cell leukemia, acute myeloid leukemia, myelodysplastic syndrome, immunoglobulin A lambda myeloma, diffuse mixed histiocytic and lymphocytic lymphoma, B-cell lymphoma, acute lymphoblastic leukemia (e.g., T-cell acute lymphoblastic leukemia or B-cell acute lymphoblastic leukemia), diffuse large cell lymphoma, or non-Hodgkin's lymphoma). In some embodiments, the cancer is breast cancer (e.g., ER-positive breast cancer, ER-negative breast cancer, triple-positive breast cancer, or triple-negative breast cancer). In some embodiments, the cancer is bone cancer (e.g., Ewing's sarcoma). In some embodiments, the cancer is renal cell carcinoma (e.g., microphthalmia transcription factor (MITF) family translocation renal cell carcinoma (tRCC)).

[0021] In some embodiments of any of the foregoing methods, the cancer is drug resistant (e.g., the cancer has been determined to be or is likely to be resistant to a chemotherapeutic or cytotoxic agent, e.g., by genetic markers, or is likely to be resistant to a chemotherapeutic or cytotoxic agent, such as a cancer that has not responded to a chemotherapeutic or cytotoxic agent), and / or has not responded to a previous therapy (e.g., a chemotherapeutic or cytotoxic agent, immunotherapy, surgery, radiation therapy, heat treatment, or photocoagulation, or a combination thereof).

[0022] In some embodiments of any of the foregoing methods, the cancer is resistant and / or non-responsive to vemurafenib, dacarbazine, CTLA4 inhibitors, PD-1 inhibitors, interferon therapy, BRAF inhibitors, MEK inhibitors, radiation therapy, temozolimide, irinotecan, CAR-T therapy, Herceptin, Perjeta, tamoxifen, Xeloda, docetaxol, platinum agents such as carboplatin, taxanes such as paclitaxel and docetaxel, ALK inhibitors, MET inhibitors, Alimta, Abraxane, doxorubicin, gemcitabine, Avastin, Halaven, neratinib, PARP inhibitors, brilanestrant, mTOR inhibitors, topotecan, gemzar, VEGFR2 inhibitors, folate receptor antagonists, demcizumab, fosbretabulin, CD-161 inhibitors, or PD-L1 inhibitors), or combinations thereof.

[0023] In some embodiments of any of the foregoing methods, the cancer is resistant to or has failed to respond to dacarbazine, temozolomide, cisplatin, treosulfan, fotemustine, IMCgp100, CTLA-4 inhibitors (e.g., ipilimumab), PD-1 inhibitors (e.g., nivolumab or pembrolizumab), PD-L1 inhibitors (e.g., atezolizumab, avelumab, or durvalumab), mitogen-activated protein kinase (MEK) inhibitors (e.g., selumetinib, binimetinib, or trametinib), and / or protein kinase C (PKC) inhibitors (e.g., sotrastaurin or IDE196).

[0024] In some embodiments of any of the foregoing methods, the cancer is resistant or unresponsive to previously administered therapeutic agents used to treat uveal melanoma, such as MEK inhibitors or PKC inhibitors. For example, in some embodiments, the cancer is resistant or unresponsive to mitogen-activated protein kinase (MEK) inhibitors (e.g., selumetinib, binimetinib, or tametinib) and / or protein kinase C (PKC) inhibitors (e.g., sotrastaurin or IDE196).

[0025] In some embodiments, the agent that reduces the level and / or activity of BRD7 in a cell is a small molecule compound, an antibody, an enzyme, and / or a polynucleotide.

[0026] In some embodiments, the agent that reduces the level and / or activity of BRD7 in cells is an enzyme.In some embodiments, the enzyme is a clustered regularly interspaced short palindromic repeats (CRISPR) associated protein, a zinc finger nuclease (ZFN), a transcription activator-like effector nuclease (TALEN) or a meganuclease.In some embodiments, the CRISPR associated protein is CRISPR associated protein 9 (Cas9).

[0027] In some embodiments, the agent that reduces the level and / or activity of BRM and / or BRG1 in a cell is an enzyme. In some embodiments, the polynucleotide is an antisense nucleic acid, a short interfering RNA (siRNA), a short hairpin RNA (shRNA), a microRNA (miRNA), a CRISPR / Cas9 nucleotide (e.g., a guide RNA (gRNA)), or a ribozyme. In some embodiments, the agent that reduces the level and / or activity of BRM and / or BRG1 in a cell is a small molecule compound (e.g., a small molecule BRM and / or BRG1 inhibitor, such as a BRM and / or BRG1 inhibitor that is selective for BRM over BRG1 or that is selective for BRG1 over BRM). In some embodiments, the small molecule compound is a degrading agent.

[0028] In some embodiments, the agent that reduces the level and / or activity of BRM and / or BRG1 is N-(1-((4-(6-(2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide, or a pharma- ceutical acceptable salt thereof, having the structure: [ka]

[0029] In some embodiments, the agent that reduces the level and / or activity of BRM and / or BRG1 has the structure, or a pharma- ceutically acceptable salt thereof: [ka]

[0030] In some embodiments, the agent that reduces the level and / or activity of BRM and / or BRG1 has the structure, or a pharma- ceutically acceptable salt thereof: [ka]

[0031] In some embodiments, the agent that reduces the level and / or activity of BRM and / or BRG1 has the structure, or a pharma- ceutically acceptable salt thereof: [ka]

[0032] In some embodiments, the methods include administering a pharmaceutical composition comprising any of the aforementioned compounds and a pharma- ceutically acceptable excipient.

[0033] In some embodiments of any of the aforementioned methods, an effective amount of a compound reduces the level and / or activity of BRG1 by at least 5% (e.g., at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%) compared to a reference substance.

[0034] In some embodiments of any of the aforementioned methods, an effective amount of a compound reduces the level and / or activity of BRG1 by at least 5% (e.g., at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%) for at least 12 hours (e.g., at least 14 hours, at least 16 hours, at least 18 hours, at least 20 hours, at least 22 hours, at least 24 hours, at least 30 hours, at least 36 hours, at least 48 hours, at least 72 hours, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 14 days, at least 21 days, at least 28 days, or more) compared to a reference substance.

[0035] In some embodiments of any of the aforementioned methods, an effective amount of a compound reduces the level and / or activity of a BRM by at least 5% (e.g., at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%) compared to a reference substance.

[0036] In some embodiments of any of the aforementioned methods, an effective amount of a compound reduces the level and / or activity of a BRM by at least 5% (e.g., at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%) for at least 12 hours (e.g., at least 14 hours, at least 16 hours, at least 18 hours, at least 20 hours, at least 22 hours, at least 24 hours, at least 30 hours, at least 36 hours, at least 48 hours, at least 72 hours, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 14 days, at least 21 days, at least 28 days, or more) compared to a reference substance.

[0037] In some embodiments, the effective amount of a compound of the invention is an amount effective to inhibit metastatic colonization of cancer to the liver and / or brain.

[0038] In another embodiment of any of the above methods, the method further comprises administering to the subject an additional anti-cancer therapy, such as a chemotherapeutic or cytotoxic agent, immunotherapy, surgery, radiation therapy, hyperthermia, or photocoagulation, or a combination thereof.In some embodiments, the anti-cancer therapy is a chemotherapeutic or cytotoxic agent, such as antimetabolites, antimitotics, antitumor agents, antibiotics, asparagine-specific enzymes, bisphosphonates, anti-neoplastic agents, alkylating agents, DNA repair enzyme inhibitors, histone deacetylase inhibitors, corticosteroids, demethylating agents, immunomodulators, Janus-related kinase inhibitors, phosphinositide 3-kinase inhibitors, proteasome inhibitors, or tyrosine kinase inhibitors, or a combination thereof.

[0039] In some embodiments, the compounds of the present invention are used in combination with another anti-cancer therapy used to treat uveal melanoma, such as surgery, MEK inhibitors, and / or PKC inhibitors, or combinations thereof. For example, in some embodiments, the method further comprises performing surgery before, after, or simultaneously with administration of the compounds of the present invention. In some embodiments, the method further comprises administering a MEK inhibitor (e.g., selumetinib, binimetinib, or tametinib) and / or a PKC inhibitor (e.g., sotrastaurin or IDE196) before, after, or simultaneously with administration of the compounds of the present invention.

[0040] In some embodiments, the anti-cancer therapy and the compound of the invention are administered within 28 days (e.g., within 21 days, within 14 days, or within 7 days) of each other, each in an amount effective together to treat the subject.

[0041] In certain embodiments, the antiproliferative agent is a chemotherapeutic or cytotoxic agent, a differentiation inducer (e.g., retinoic acid, vitamin D, cytokines), a hormonal agent, an immunological agent, or an antiangiogenic agent. Chemotherapeutic and cytotoxic agents include, but are not limited to, alkylating agents, cytotoxic antibiotics, antimetabolites, vinca alkaloids, etoposide, and other agents (e.g., paclitaxel, taxol, docetaxel, taxotere, cis-platin). A list of additional compounds with antiproliferative agents can be found in L. Brunton, B. Chabner and B. Knollman (eds). Goodman and Gilman's The Pharmacological Basis of Therapeutics, Twelfth Edition, 2011, McGraw Hill Companies, New York, NY.

[0042] The method includes the use of alkylating agents, platinum agents, antimetabolites, topoisomerase inhibitors, antitumor antibiotics, antimitotic agents, aromatase inhibitors, thymidylate synthase inhibitors, DNA antagonists, farnesyltransferase inhibitors, pump inhibitors, histone acetyltransferase inhibitors, metalloproteinase inhibitors, ribonucleoside reductase inhibitors, TNF The method may further comprise administering an anti-proliferative agent selected from the group consisting of alpha agonists / antagonists, endothelin A receptor antagonists, retinoic acid receptor agonists, immunomodulatory agents, hormonal and anti-hormonal agents, photodynamic agents, tyrosine kinase inhibitors, antisense compounds, corticosteroids, HSP90 inhibitors, proteosome inhibitors (e.g., NPI-0052), CD40 inhibitors, anti-CSI antibodies, FGFR3 inhibitors, VEGF inhibitors, MEK inhibitors, cyclin D1 inhibitors, NF-kB inhibitors, anthracyclines, histone deacetylases, kinesin inhibitors, phosphatase inhibitors, COX2 inhibitors, mTOR inhibitors, calcineurin antagonists, IMiDs, or other agents used to treat proliferative diseases.

[0043] In certain embodiments, the antiproliferative agent and / or immunotherapy, and the agent that reduces the level and / or activity of BRM and / or BRG1 are administered within 28 days (e.g., within 21, 14, 10, 7, 5, 4, 3, 2, or 1 days) or within 24 hours (e.g., within 12, 6, 3, 2, or 1 hours, or simultaneously) of each other, in amounts each effective to treat the subject.

[0044] In another aspect, the invention features an agent that reduces the level and / or activity of BRM and / or BRG1 (e.g., an agent described herein) for use in combination with immunotherapy (e.g., an immunotherapy described herein), e.g., according to the methods described herein, to treat cancer (e.g., a cancer described herein) in a subject in need of such treatment.

[0045] In yet another aspect, the invention provides a compound having the structure: [ka] or a pharma- ceutically acceptable salt thereof.

[0046] In another aspect, the invention features the use of an agent that reduces the level and / or activity of BRM and / or BRG1 (e.g., an agent described herein) in combination with an immunotherapy (e.g., an immunotherapy described herein), e.g., according to the methods described herein, in the manufacture of a medicament, to treat cancer (e.g., a cancer described herein) in a subject in need thereof.

[0047] In yet another aspect, the invention provides a compound having the structure: [ka] or a pharma- ceutically acceptable salt thereof.

[0048] chemical terms The compounds of the present invention may have one or more asymmetric carbon atoms and may exist in the form of optically pure enantiomers, mixtures of enantiomers, such as racemates, optically pure diastereomers, mixtures of diastereomers, diastereomeric racemates, or mixtures of diastereomeric racemates. Optically active forms can be obtained, for example, by resolution of racemates, by asymmetric synthesis or asymmetric chromatography (chromatography using chiral adsorbents or eluents). That is, certain disclosed compounds may exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial arrangement. Enantiomers are most commonly pairs of stereoisomers whose mirror images are not superimposable because they contain asymmetrically substituted carbon atoms that function as chiral centers. Enantiomers refer to one of a pair of molecules that are mirror images of each other and are not superimposable. Diastereomers are most commonly stereoisomers that are not related as mirror images because they contain two or more asymmetrically substituted carbon atoms and represent the configuration of substituents around one or more chiral carbon atoms. Enantiomers of a compound may be prepared, for example, by separating an enantiomer from a racemate using one or more well-known techniques and methods, such as, for example, chiral chromatography and separation methods based thereon. Suitable techniques and / or methods for separating enantiomers of the compounds described herein from racemic mixtures can be readily determined by one of ordinary skill in the art. "Racemate" or "racemic mixture" refers to a compound containing two enantiomers, and such mixtures do not exhibit optical activity, i.e., they do not rotate the plane of polarized light. "Geometric isomer" refers to isomers that differ in the orientation of substituted atoms in relationship to a carbon-carbon double bond, a cycloalkyl ring, or a bridged bicyclic system. Atoms (other than H) on each side of a carbon-carbon double bond may be in the E (substituents are on opposite sides of the carbon-carbon double bond) or Z (substituents are oriented on the same side) configuration. "R", "S", "S*", "R*", "E", "Z", "cis", and "trans" refer to configurations relative to the core molecule. Certain disclosed compounds may exist in atropisomeric forms.Atropisomers are stereoisomers resulting from hindered rotation around a single bond, where the steric strain barrier to rotation is high enough to allow the isolation of the conformers. The compounds of the present invention can be prepared as individual isomers by isomer-specific synthesis or by resolution from an isomeric mixture. Traditional resolution techniques include forming a salt of the free base of each isomer of the isomeric pair using an optically active acid (followed by fractional crystallization and regeneration of the free base), forming a salt of the acid form of each isomer of the isomeric pair using an optically active amine (followed by fractional crystallization and regeneration of the free acid), forming an ester or amide of each isomer of the isomeric pair using an optically pure acid, amine, or alcohol (followed by chromatographic separation and removal of the chiral auxiliary), or resolving the isomeric mixture of either the starting material or the final product using a variety of well-known chromatographic methods. When the stereochemistry of a disclosed compound is named or shown by structure, the named or shown stereoisomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight relative to the other stereoisomers. When a single enantiomer is named or shown by structure, the shown or named enantiomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% optically pure by weight. When a single diastereomer is named or shown by structure, the shown or named diastereomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by weight. Percent optical purity is the weight of the enantiomer, or the ratio of the weight of the enantiomer to the weight of its optical isomer. Diastereomeric purity by weight is the ratio of the weight of one diastereomer to the weight of all diastereomers. When the stereochemistry of a disclosed compound is named or shown by structure, the named or shown stereoisomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by mole fraction relative to the other stereoisomer.When a single enantiomer is named or shown by structure, the shown or named enantiomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by mole fraction. When a single diastereomer is named or shown by structure, the shown or named diastereomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by mole fraction. Percent purity by mole fraction is the ratio of moles of enantiomer, or moles of enantiomer and moles of its optical isomer. Similarly, percent purity by mole fraction is the ratio of moles of diastereomer, or moles of diastereomer and moles of its isomer. When a disclosed compound is named or shown by structure without showing stereochemistry and the compound has at least one chiral center, the name or structure should be understood to encompass any of the enantiomers of the compound without the corresponding optical isomer, a racemic mixture of the compound, a mixture of the compound, or a mixture enriched in one enantiomer with respect to its corresponding optical isomer. When a disclosed compound is named or shown by structure without showing stereochemistry and has two or more chiral centers, the name or structure should be understood to encompass any of the diastereomers without the other diastereomers, some diastereomers without the other diastereomeric pairs, mixtures of diastereomers, mixtures of diastereomeric pairs, mixtures of diastereomers enriched in one diastereomer with respect to the other diastereomer, or mixtures of diastereomers enriched in one or more diastereomers with respect to the other diastereomer(s). The present invention encompasses all of these forms.

[0049] Unless otherwise stated, structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. Exemplary isotopes that can be incorporated into the compounds of the invention include: 2 H, 3 H, 11 C. 13 C. 14 C. 13 N,15 N, 15 O. 17 O. 18 O. 32 P, 33 P, 35 S, 18 F, 36 Cl, 123 I, and 125 Isotopically labeled compounds (e.g., 3 H and 14 C) can be useful in compound or substrate tissue distribution assays. 3 H) and carbon-14 (i.e. 14 C) isotopes can be useful for their ease of preparation and detectability. Additionally, deuterium (i.e., 2 Substitution with heavier isotopes, such as H, may confer certain therapeutic advantages (e.g., increased in vivo half-life or reduced dosage requirements) due to greater metabolic stability. In some embodiments, one or more hydrogen atoms are 2 H or 3 H or one or more carbon atoms are replaced by 13 C or 14 Replaced by C-enriched carbon. 15 O. 13 N, 11 C, and 18 Positron-emitting isotopes such as F are useful for positron emission tomography (PET) studies to examine substrate receptor occupancy. Preparation of isotopically labeled compounds is known to those skilled in the art. For example, isotopically labeled compounds can generally be prepared by following procedures similar to those disclosed for the compounds of the present invention described herein, by substituting isotopically labeled reagents with non-isotopically labeled reagents.

[0050] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. Methods and materials for use in this disclosure are described herein. Other suitable methods and materials known in the art may also be used. These materials, methods, and examples are illustrative only and are not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

[0051] definition In this application, unless otherwise clear from the context, (i) the term "a" may be understood to mean "at least one," (ii) the term "or" may be understood to mean "and / or," and (iii) the terms "comprising" and "including" may be understood to encompass the itemized elements or steps, whether presented by themselves or with one or more additional elements or steps.

[0052] As used herein, the term "A1 adenosine inhibitor" refers to a compound, such as an antibody, capable of inhibiting the activity of the protein encoded by the ADORA1 gene (Accession No. P30542) in humans. Known A1 adenosine inhibitors include 8-cyclopentyl-1,3-dimethylxanthine, 8-cyclopentyl-1,3-dipropylxanthine, 8-phenyl-1,3-dipropylxanthine, bamifylline, BG-9719, BG-9928, FK-453, FK-838, rolofylline, and N-0861.

[0053] As used herein, the term "A2A adenosine inhibitor" refers to a compound, such as an antibody, that can inhibit the activity of the protein encoded by the ADORA2A gene (Accession No. P29274) in humans. Known A2A adenosine inhibitors include ATL-4444, istradefylline, MSX-3, preladenant, SCH-58261, SCH-412,348, SCH-442,416, ST-1535, VER-6623, VER-6947, VER-7835, viadenant, and ZM-241,385.

[0054] As used herein, the term "A2B adenosine inhibitor" refers to a compound, such as an antibody, that can inhibit the activity of the protein encoded by the ADORA2B gene (Accession No. P29275) in humans. Known A2B adenosine inhibitors include ATL-801, CVT-6883, MRS-1706, MRS-1754, OSIP-339,391, PSB-603, PSB-0788, and PSB-1115.

[0055] As used herein, the term "A3A adenosine inhibitor" refers to a compound, such as an antibody, that can inhibit the activity of the protein encoded by the ADORA3 gene (Accession No. P0DMS8) in humans. Known A3A adenosine inhibitors include KF-26777, MRS-545, MRS-1191, MRS-1220, MRS-1334, MRS-1523, MRS-3777, MRE-3005-F20, MRE-3008-F20, PSB-11, OT-7999, VUF-5574, and SSR161421.

[0056] As used herein, the terms "about" and "approximately" refer to values ​​within 10% above or below the stated value. For example, the term "about 5 nM" indicates a range of 4.5 to 5.5 nM.

[0057] As used herein, the term "administration" refers to administration of a composition (e.g., a compound or a preparation comprising a compound as described herein) to a subject or system. Administration to an animal subject (e.g., to a human) can be by any suitable route. For example, in some embodiments, administration can be bronchial (including by bronchial instillation), buccal, enteral, interdermal, intraarterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intratumoral, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal, and vitreous.

[0058] As used herein, the term "arginase inhibitor" refers to a compound capable of inhibiting the activity of the protein encoded by the ARG1 gene (Accession No. P05089) or the ARG2 gene (Accession No. P78540) in humans. Known arginase inhibitors include (2s)-(+)-amino-5-iodoacetamidopentanoic acid, NG-hydroxy-L-arginine, (2S)-(+)-amino-6-iodoacetamidohexanoic acid, and (R)-2-amino-6-borono-2-(2-(piperidin-1-yl)ethyl)hexanoic acid.

[0059] As used herein, the term "BAF complex" refers to the BRG1 or HBRM-associated factor complex in human cells.

[0060] As used herein, the term "BAF complex-associated disorder" refers to a disorder caused by or affected by the level of activity of the BAF complex.

[0061] As used herein, the term "loss-of-function mutation of BRG1" refers to a mutation in BRG1 that results in a protein with reduced activity (e.g., at least a 1% reduction in BRG1 activity, e.g., a 2%, 5%, 10%, 25%, 50%, or 100% reduction in BRG1 activity). Exemplary BRG1 loss-of-function mutations include, but are not limited to, homozygous BRG1 mutations and deletions at the C-terminus of BRG1.

[0062] As used herein, the term "BRG1 loss-of-function disorder" refers to a disorder (e.g., cancer) that exhibits a reduction in BRG1 activity (e.g., at least a 1% reduction in BRG1 activity, e.g., a 2%, 5%, 10%, 25%, 50%, or 100% reduction in BRG1 activity).

[0063] The term "cancer" refers to conditions caused by the proliferation of malignant cells, such as tumors, neoplasms, carcinomas, sarcomas, leukemias, and lymphomas.

[0064] As used herein, "combination therapy" or "administered in combination" means that two (or more) different agents or treatments are administered to a subject as part of a defined treatment regimen for a particular disease or condition. The treatment regimen defines the dose and periodicity of administration of each agent such that the effects of the separate agents on the subject overlap. In some embodiments, the delivery of the two or more agents may be simultaneous or parallel, and the agents may be co-formulated. In some embodiments, the two or more agents are not co-formulated and are administered in a sequential manner as part of a prescribed regimen. In some embodiments, the administration of the two or more agents or combined treatments is such that the reduction in symptoms, or other parameters associated with the disorder, is greater than that observed with one agent or treatment delivered alone or in the absence of the other. The effect of the two treatments may be partially additive, fully additive, or greater than additive (e.g., synergistic). The sequential or substantially simultaneous administration of each therapeutic agent may occur by any suitable route, including, but not limited to, oral, intravenous, intramuscular, and direct absorption through mucosal tissue. The therapeutic agents can be administered by the same route or by different routes, for example, a first therapeutic agent of the combination may be administered by intravenous injection, while a second therapeutic agent of the combination may be administered orally.

[0065] As used herein, the term "CTLA-4 inhibitor" refers to a compound, such as an antibody, that can inhibit the activity of the protein encoded by the CTLA4 gene in humans. Known CTLA-4 inhibitors include ipilimumab.

[0066] As used herein, the term "CSF-1R inhibitor" refers to a compound, such as an antibody, capable of inhibiting the activity of the protein encoded by the CSF1R gene (Accession No. P07333) in humans. Known CSF-1R inhibitors include pexidartinib and AZD6495.

[0067] "Determining the level" of a protein or RNA refers to detection of the protein or RNA, either directly or indirectly, by methods known in the art. "Directly determining" means performing a process to obtain a physical entity or value (e.g., performing an assay or test on a sample, or "analyzing a sample" as that term is defined herein). "Indirectly determining" refers to receiving a physical entity or value from another party or source (e.g., a third party laboratory that obtains the physical entity or value directly). Methods for measuring protein levels generally include, but are not limited to, Western blotting, immunoblotting, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunoprecipitation, immunofluorescence, surface plasmon resonance, chemiluminescence, fluorescence polarization, phosphorescence, immunohistochemistry, matrix-assisted laser desorption / ionization time-of-flight (MALDI-TOF) mass spectrometry, liquid chromatography (LC) mass spectrometry, microcytometry, microscopy, fluorescence-activated cell sorting (FACS), and flow cytometry, as well as assays based on the properties of the protein, including, but not limited to, enzyme activity or interaction with other protein partners. Methods for measuring RNA levels are known in the art and include, but are not limited to, quantitative polymerase chain reaction (qPCR) and Northern blot analysis.

[0068] By "decreased levels" or "increased levels" of protein or RNA is meant a decrease or increase, respectively, in protein or RNA levels compared to a reference material (e.g., a decrease or increase of about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, or more). or a decrease or increase of about 10%, about 15%, about 20%, about 50%, about 75%, about 100%, or more than about 200% compared to a reference material, or a decrease or increase of less than about 0.01-fold, about 0.02-fold, about 0.1-fold, about 0.3-fold, about 0.5-fold, about 0.8-fold, or less, or an increase of about 1.2-fold, about 1.4-fold, about 1.5-fold, about 1.8-fold, about 2.0-fold, about 3.0-fold, about 3.5-fold, about 4.5-fold, about 5.0-fold, about 10-fold, about 15-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 100-fold, about 1000-fold, or more. The level of protein may be expressed as mass / volume (e.g., g / dL, mg / mL, μg / mL, ng / mL) or as a percentage of the total protein in the sample.

[0069] "Reduce the activity of BAF complex" means to reduce the level of activity related to BAF complex or the downstream effect related.A non-limiting example of reducing the activity of BAF complex is the activation of Sox2.The activity level of BAF complex can be measured using any method known in the art, for example, the method described in Kadoch et al.Cell,2013,153,71-85, which is incorporated herein by reference.

[0070] As used herein, the term "derivative" refers to naturally occurring, synthetic, and semi-synthetic analogs of the compounds, peptides, proteins, or other substances described herein. Derivatives of the compounds, peptides, proteins, or other substances described herein may retain or improve the biological activity of the original material.

[0071] As used herein, a cancer "determined to be drug resistant" refers to a cancer that is drug resistant based on no response or reduced responsiveness to chemotherapeutic agents, or that is predicted to be drug resistant based on a prognostic assay (e.g., a gene expression assay).

[0072] By "drug resistance" is meant a cancer that is unresponsive or shows a reduced response to one or more chemotherapeutic agents (eg, any of the agents described herein).

[0073] As used herein, the terms "failed to respond to previous treatment" or "ineffective to previous treatment" refer to cancer that has progressed despite treatment with a therapy.

[0074] As used herein, the term "HDAC inhibitor" refers to a compound, such as an antibody, that can inhibit the activity of proteins that are members of the histone deacetylase class of enzymes, e.g., HDAC1, HDAC2, HDAC3, HDAC4, HDAC5, HDAC6, HDAC7, HDAC8, HDAC9, HDAC10, HDAC11, SIRT1, SIRT2, SIRT3, SIRT4, SIRT5, SIRT6, and SIRT7. Known HDAC inhibitors include valproic acid, SAHA, and romidepsin.

[0075] As used herein, the term "IDO inhibitor" refers to a compound, such as an antibody, capable of inhibiting the activity of the protein encoded by the IDO1 gene (Accession No. P14902) in humans. Known IDO inhibitors include norharman, rosmarinic acid, and α-methyl-tryptophan.

[0076] As used herein, the terms "inhibiting BRM" and / or "inhibiting BRG1" refer to blocking or reducing the level or activity of the ATPase catalytic binding domain or bromodomain of a protein. BRM and / or BRG1 inhibition can be determined using methods known in the art, such as a BRM and / or BRG1 ATPase assay, a Nano DSF assay, or a BRM and / or BRG1 luciferase cellular assay.

[0077] As used herein, the term "LXS196," also known as IDE196, refers to a PKC inhibitor having the following structure: [ka] or a pharma- ceutically acceptable salt thereof.

[0078] As used herein, "metastatic nodule" refers to an aggregation of tumor cells in the body at a site other than the site of the original tumor.

[0079] As used herein, "metastatic cancer" refers to a tumor or cancer in which the cancer cells forming the tumor have a high likelihood of or have begun to metastasize or spread from one location to another location(s) within a subject, e.g., giving rise to a secondary tumor within the subject, via the lymphatic system or via blood-borne spread. Such metastatic behavior may be indicative of a malignant tumor. In some cases, metastatic behavior may be associated with increased cell migration and / or invasive behavior of the tumor cells.

[0080] Examples of cancers that may be defined as metastatic include, but are not limited to, lung cancer (e.g., non-small cell lung cancer), breast cancer, ovarian cancer, colon cancer, biliary tract cancer, bladder cancer, brain cancer including glioblastoma and medulloblastoma, cervical cancer, choriocarcinoma, endometrial cancer, esophageal cancer, gastric cancer, hematological neoplasms, multiple myeloma, leukemia, intraepithelial neoplasia, liver cancer, lymphoma, neuroblastoma, oral cancer, pancreatic cancer, prostate cancer, sarcoma, skin cancer including melanoma, basal cell carcinoma, squamous cell carcinoma, testicular cancer, stromal tumors, germ cell tumors, thyroid cancer, and renal cancer.

[0081] As used herein, "non-metastatic cell migration cancer" refers to a cancer that does not migrate via the lymphatic system or via hematogenous spread.

[0082] As used herein, the term "PD-1 inhibitor" refers to a compound, such as an antibody, that can inhibit the activity of a protein encoded by the PDCD1 gene in humans. Known PD-1 inhibitors include nivolumab, pembrolizumab, pidilizumab, BMS 936559, and atezolizumab.

[0083] As used herein, the term "PD-L1 inhibitor" refers to a compound, such as an antibody, that can inhibit the activity of a protein encoded by the CD274 gene in humans. Known PD-L1 inhibitors include atezolizumab and durvalumab.

[0084] As used herein, the term "pharmaceutical composition" refers to a composition comprising a compound described herein, formulated with a pharmaceutically acceptable excipient and suitable for administration to a mammal, e.g., a human. Typically, a pharmaceutical composition is manufactured or sold by approval of a government regulatory agency as part of a therapeutic regimen for the treatment of a mammalian disease. A pharmaceutical composition can be formulated, for example, for oral administration in a unit dosage form (e.g., tablet, capsule, caplet, gelcap, or syrup), for topical administration (e.g., as a cream, gel, lotion, or ointment), for intravenous administration (e.g., as a sterile solution free of particulate embolic material and in a solvent system suitable for intravenous use), or in any other pharmaceutically acceptable formulation.

[0085] As used herein, "pharmaceutical acceptable excipient" refers to any component other than the compound described herein (e.g., a vehicle capable of suspending or dissolving an active compound) and having the properties of being substantially non-toxic and non-inflammatory in a patient. Excipients may include, for example, anti-adherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film-forming or coating agents, flavors, fragrances, glidants (glidants), lubricants, preservatives, printing inks, adsorbents, suspending or dispersing agents, sweeteners, and water of hydration.

[0086] As used herein, the term "pharmaceutically acceptable salt" refers to any pharmaceutically acceptable salt of the compounds described herein. Any pharmaceutically acceptable salt of the compounds described herein may include salts that are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, or allergic response, within the scope of sound medical judgment, and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. Salts can be prepared in situ during the final isolation and purification of the compounds described herein, or separately by reacting the free base group with a suitable organic acid.

[0087] The compounds of the present invention may have ionizable groups so that they can be prepared as pharmaceutically acceptable salts.These salts may be acid addition salts, including inorganic or organic acids, or salts may be prepared from inorganic or organic bases when the compounds of the present invention are in acidic form.Frequently, the compounds are prepared or used as pharmaceutically acceptable salts, which are prepared as addition products of pharmaceutically acceptable acids or bases.Suitable pharmaceutically acceptable acids and bases, and the preparation method of suitable salts are well known in the art.Salts may be prepared from pharmaceutically acceptable non-toxic acids and bases, including inorganic and organic acids and bases.

[0088] As used herein, "progression-free survival" refers to the length of time during and after a medication or treatment during which the disease being treated (e.g., cancer) does not worsen.

[0089] As used in this application, "proliferation" involves the duplication or proliferation of similar morphologies (cells) resulting from constituting (cellular) elements.

[0090] "Reducing the activity of BRM and / or BRG1" means decreasing the level of activity or downstream effects associated with BRM and / or BRG1. The activity level of BRM and / or BRG1 may be measured using any method known in the art. In some embodiments, the agent that reduces the activity of BRM and / or BRG1 is a small molecule BRM and / or BRG1 inhibitor. In some embodiments, the agent that reduces the activity of BRM and / or BRG1 is a small molecule BRM and / or BRG1 degrader.

[0091] "Reducing the level of BRM and / or BRG1" means reducing the level of BRM and / or BRG1 in a cell or a subject, for example, by administering a degrading agent to the cell or subject. The level of BRM and / or BRG1 can be measured using any method known in the art.

[0092] "Reference material" refers to any useful reference material used to compare protein or RNA levels. A reference material can be any sample, standard, standard curve, or level used for comparison purposes. A reference material can be a normal reference sample or a reference standard or level. A "reference sample" can be, for example, a control, e.g., a predefined negative control value such as a "normal control," or a previous sample taken from the same subject; a sample from a normal healthy subject, such as a normal cell or normal tissue; a sample (e.g., cell or tissue) from a subject without a disease; a sample from a subject diagnosed with a disease but not yet treated with a compound of the invention; a sample from a subject being treated with a compound of the invention; or a sample of a known normal concentration of purified protein or RNA (e.g., any of those described herein). "Reference standard or level" refers to a value or numerical value derived from a reference sample. A "normal control value" is a predefined value indicative of a non-disease state, e.g., a value expected in a healthy control subject. Typically, a normal control value is expressed as a range ("between X and Y"), a high threshold ("below X"), or a low threshold ("above X"). A subject having a measurement value within the normal control value of a particular biomarker is typically referred to as being "within the normal range" of that biomarker. A normal reference standard or level can be a value or number derived from a normal subject without a disease or disorder (e.g., cancer); a subject being treated with a compound of the present invention. In a preferred embodiment, the reference sample, standard, or level is matched to the sample subject sample by at least one of the following criteria: age, weight, sex, stage of disease, and overall health. A standard curve of purified protein or RNA levels within the normal reference range, such as any of those described herein, can also be used as a reference.

[0093] As used herein, the term "selective for a BRM over BRG1" refers to a compound that inhibits the level and / or activity of a BRM by at least 5% (e.g., at least 10%, at least 25%, at least 50%, at least 75%, or at least 100%) more than the compound inhibits the level and / or activity of BRG1.

[0094] As used herein, the term "selective for BRG1 over BRM" refers to a compound that inhibits the level and / or activity of BRG1 by at least 5% (e.g., at least 10%, at least 25%, at least 50%, at least 75%, or at least 100%) more than the compound inhibits the level and / or activity of BRM.

[0095] As used herein, "slowing the spread of metastases" refers to reducing or stopping the formation of new loci or reducing, stopping or reversing the tumor burden.

[0096] As used herein, the term "subject" refers to any organism to which a composition according to the invention may be administered, for example, for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Typical subjects include any animal (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans). A subject may be a human or animal seeking or in need of treatment, requesting treatment, undergoing treatment, will be undergoing treatment in the future, or being treated by a trained professional for a particular disease or condition.

[0097] As used herein, the terms "treat", "treated" or "treating" refer to a therapeutic treatment or any procedure, the purpose of which is to slow (alleviate) an undesirable physiological condition, disorder, or disease, or to obtain a beneficial or desired clinical outcome. Beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms; attenuation of the extent of the condition, disorder, or disease; stabilization (i.e., not worsening) of the condition, disorder, or disease; delaying or slowing the onset of the progression of the condition, disorder, or disease; improvement or remission (partial or complete) of the condition, disorder, or disease state; amelioration of at least one measurable physical parameter, not necessarily discernible by the patient; or enhancement or improvement of the condition, disorder, or disease. Treatment includes eliciting a clinically significant response without an excessive level of side effects. Treatment also includes extending survival compared to the expected survival in the absence of treatment. The compounds of the invention can also be used to "prophylactically treat" or "prevent" disorders, for example, in subjects at increased risk of developing the disorder.

[0098] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. Methods and materials for use in this disclosure are described herein. Other suitable methods and materials known in the art may also be used. These materials, methods, and examples are illustrative only and are not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

[0099] The details of one or more embodiments of the invention are set forth in the description below. Other features, objects, and advantages of the invention will become apparent from the description and the claims. [Brief description of the drawings]

[0100] [Figure 1] 1 is a graph showing inhibition of cell proliferation of several cancer cell lines by a BRG1 / BRM inhibitor (Compound A). [Diagram 2] 1 is a graph showing inhibition of cell proliferation of uveal melanoma cell line 92-1 by a BRG1 / BRM inhibitor (Compound A), a MEK inhibitor (selumetinib), and a PKC inhibitor (LXS196). [Diagram 3] 1 is a graph showing inhibition of cell proliferation of the uveal melanoma cell line MP41 by a BRG1 / BRM inhibitor (Compound A), a MEK inhibitor (selumetinib), and a PKC inhibitor (LXS196). [Figure 4] 1 is a graph showing inhibition of cell proliferation of several cancer cell lines by a BRG1 / BRM inhibitor (Compound B). [Diagram 5] FIG. 1 is a graph showing the area under the curve (AUC) calculated from the dose-response curve of cancer cell lines treated with a BRG1 / BRM inhibitor (Compound B). [Figure 6] 1 is a graph showing inhibition of cell proliferation of uveal melanoma and non-small cell lung cancer cell lines by a BRG1 / BRM inhibitor (Compound B). [Figure 7] 1 is a graph showing inhibition of cell proliferation of uveal melanoma cell line 92-1 by a BRG1 / BRM inhibitor (Compound B), a MEK inhibitor (selumetinib), and a PKC inhibitor (LXS196). [Figure 8] 1 is a graph showing inhibition of cell proliferation of the uveal melanoma cell line MP41 by a BRG1 / BRM inhibitor (Compound B), a MEK inhibitor (selumetinib), and a PKC inhibitor (LXS196). [Figure 9] 1 is a graph showing inhibition of cell proliferation of parental and PKC inhibitor-refractory uveal melanoma cell lines by a PKC inhibitor (LXS196). [Figure 10] 1 is a graph showing inhibition of cell proliferation of parental and PKC inhibitor-refractory uveal melanoma cell lines by a BRG1 / BRM inhibitor (Compound B). [Figure 11] 1 is a graph showing inhibition of tumor growth in mice implanted with uveal melanoma cell lines by a BRG1 / BRM inhibitor (Compound C). [Figure 12] FIG. 1 shows the size of tumors from mice implanted with uveal melanoma cell lines and administered a BRG1 / BRM inhibitor (Compound C). [Figure 13] 1 is a graph showing changes in body weight of mice transplanted with a uveal melanoma cell line and administered a BRG1 / BRM inhibitor (Compound C). [Figure 14] FIG. 1 is a graph showing inhibition of cell proliferation of several uveal melanoma cell lines by N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide. [Figure 15] FIG. 1 is a graph showing inhibition of tumor growth in mice implanted with uveal melanoma cell lines by N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide. [Figure 16] 1 is a graph showing the change in body weight of mice administered N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide after transplantation with a uveal melanoma cell line. [Figure 17] Graph showing tumor growth inhibition in a B16 / F10 syngeneic model with a PD-1 inhibitor alone, a BRM / BRG1 inhibitor alone, and a combination of a PD-1 inhibitor and a BRM / BRG1 inhibitor. [Figure 18] Graph showing tumor growth inhibition in a B16 / F10 syngeneic model for each individual mouse with a PD-1 inhibitor alone, a BRM / BRG1 inhibitor alone, and a combination of a PD-1 inhibitor and a BRM / BRG1 inhibitor. [Figure 19] Graph showing Kaplan-Meier survival curves in the B16 / F10 syngeneic model for each individual mouse with PD-1 inhibitor alone, BRM / BRG1 inhibitor alone, and the combination of PD-1 inhibitor and BRM / BRG1 inhibitor. [Figure 20] 1 is a graph showing tumor growth inhibition in an A20 lymphoma model with a PD-1 inhibitor alone, a BRM / BRG1 inhibitor alone, and a combination of a PD-1 inhibitor and a BRM / BRG1 inhibitor. [Figure 21] Graph showing tumor growth inhibition in the A20 lymphoma model for each individual mouse with a PD-1 inhibitor alone, a BRM / BRG1 inhibitor alone, and a combination of a PD-1 inhibitor and a BRM / BRG1 inhibitor. [Figure 22] Graph showing Kaplan-Meier survival curves in the A20 lymphoma model for each individual mouse with PD-1 inhibitor alone, BRM / BRG1 inhibitor alone, and the combination of PD-1 inhibitor and BRM / BRG1 inhibitor. [Figure 23] FIG. 1 is a graph showing tumor growth inhibition in a CT26 colorectal model with a PD-1 inhibitor alone, a BRM / BRG1 inhibitor alone, and a combination of a PD-1 inhibitor and a BRM / BRG1 inhibitor. [Figure 24] 1 is a graph showing tumor growth inhibition in a CT26 colorectal model for each individual mouse with a PD-1 inhibitor alone, a BRM / BRG1 inhibitor alone, and a combination of a PD-1 inhibitor and a BRM / BRG1 inhibitor. [Diagram 25] Graph showing Kaplan-Meier survival curves in the CT26 colorectal model for each individual mouse with PD-1 inhibitor alone, BRM / BRG1 inhibitor alone, and the combination of PD-1 inhibitor and BRM / BRG1 inhibitor. [Figure 26] Graph showing tumor growth inhibition in a CT26 colorectal model for each individual mouse with a PD-L1 inhibitor alone, a BRM / BRG1 inhibitor alone, and a combination of a PD-L1 inhibitor and a BRM / BRG1 inhibitor. [Figure 27] Graph showing Kaplan-Meier survival curves in the CT26 colorectal model for each individual mouse with PD-L1 inhibitor alone, BRM / BRG1 inhibitor alone, and combination of PD-L1 inhibitor and BRM / BRG1 inhibitor. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0101] The present inventors have found that inhibiting or depleting the levels and / or activity of BRM and / or BRG1 in cells, in combination with immunotherapy treatment, is effective in treating cancer. Thus, the present invention features, for example, a useful method for treating cancer in a subject in need thereof.

[0102] BRM and / or BRG1 reducing agent The agents described herein that reduce the level and / or activity of BRM and / or BRG1 in a cell can be antibodies, proteins (such as enzymes), polynucleotides, or small molecule compounds. The agents reduce the level of an activity associated with BRM and / or BRG1, or an associated downstream effect, or reduce the level of BRM and / or BRG1 in a cell or subject.

[0103] In some embodiments, the agent that reduces the level and / or activity of BRM and / or BRG1 in a cell is an enzyme, a polynucleotide, or a small molecule compound, e.g., a degrading agent, or a small molecule BRM and / or BRG1 inhibitor.

[0104] antibody An agent that reduces the level and / or activity of BRM and / or BRG1 can be an antibody or an antigen-binding fragment thereof. For example, an agent that reduces the level and / or activity of BRM and / or BRG1 described herein is an antibody that reduces or blocks the activity and / or function of BRM and / or BRG1 through binding to BRM and / or BRG1.

[0105] The production and use of therapeutic antibodies against target antigens (e.g., BRM and / or BRG1) are known in the art. See, for example, the references cited herein above, and Zhiqiang An (Editor), Therapeutic Monoclonal Antibodies: From Bench to Clinic. 1st Edition. Wiley 2009, and also Greenfield (Ed.), Antibodies: A Laboratory Manual. (Second edition) Cold Spring Harbor Laboratory Press 2013 for methods of producing recombinant antibodies, including antibody engineering, use of degenerate oligonucleotides, 5'-RACE, phage display, and mutagenesis, antibody testing and characterization, antibody pharmacokinetics and pharmacodynamics, antibody purification and storage, and screening and labeling techniques.

[0106] Polynucleotides In some embodiments, the agent that reduces the level and / or activity of BRM and / or BRG1 is a polynucleotide. In some embodiments, the polynucleotide is an inhibitory RNA molecule that acts, for example, by the RNA interference (RNAi) pathway. The inhibitory RNA molecule can reduce the expression level (e.g., protein level or mRNA level) of BRM and / or BRG1. For example, the inhibitory RNA molecule includes small interfering RNA (siRNA), small hairpin RNA (shRNA), and / or microRNA (miRNA) that target full-length BRM and / or BRG1. siRNA is a double-stranded RNA molecule that typically has a length of about 19-25 base pairs. shRNA is an RNA molecule that contains a hairpin turn and reduces the expression of a target gene via RNAi. MicroRNA is a non-coding RNA molecule that typically has a length of about 22 nucleotides. MiRNA binds to a target site on an mRNA molecule and silences the mRNA, for example, by causing cleavage of the mRNA, destabilization of the mRNA, or inhibition of translation of the mRNA. Degradation is triggered by the enzymatic RNA-induced silencing complex (RISC).

[0107] In some embodiments, the agent that reduces the level and / or activity of BRM and / or BRG1 is an antisense nucleic acid. Antisense nucleic acids include antisense RNA (asRNA) and antisense DNA (asDNA) molecules (typically about 10-30 nucleotides in length), which recognize a polynucleotide target sequence or sequence portion through hydrogen bonding interactions with the nucleotide bases of the target sequence (e.g., BRM and / or BRG1). The target sequence can be single- or double-stranded RNA, or single- or double-stranded DNA.

[0108] In some embodiments, the polynucleotide reduces the level and / or activity of a negative regulator of function or a positive regulator of function, hi other embodiments, the polynucleotide reduces the level and / or activity of an inhibitor of a positive regulator of function.

[0109] Polynucleotides can be modified to contain, for example, modified nucleotides (e.g., 2'-fluoro, 2'-o-methyl, 2'-deoxy, unlocked nucleic acid, 2'-hydroxy, phosphorothioate, 2'-thiouridine, 4'-thiouridine, 2'-deoxyuridine). Without being bound by theory, it is believed that certain modifications can increase nuclease resistance and / or serum stability or reduce immunogenicity. The polynucleotides can also be provided in specialized forms such as liposomes, microspheres, or for gene therapy applications, or can be provided in combination with binding moieties. Such binding moieties include polycations that act as charge neutralizers for the phosphate backbone, or lipids (e.g., liposomes) that enhance interaction with cell membranes or increase uptake of nucleic acids. Examples of suitable polynucleotides include hydrophobic moieties such as phospholipids, cholesterol, etc. These moieties may be attached to the nucleic acid at the 3' or 5' end, and may be attached via a base, sugar, or intramolecular nucleoside bond. Other moieties may be capping groups specifically placed at the 3' or 5' end of the nucleic acid to prevent degradation by nucleases such as exonucleases, RNases, etc. Such capping groups include hydroxyl protecting groups known in the art, including glycols such as polyethylene glycol and tetraethylene glycol. The inhibitory effects of polynucleotides can be tested in vivo and in vitro using the cell line or animal-based gene expression systems of the invention.

[0110] In some embodiments, the polynucleotide reduces the level and / or activity or function of BRM and / or BRG1. In embodiments, the polynucleotide inhibits expression of BRM and / or BRG1. In other embodiments, the polynucleotide increases the degradation of BRD7 and / or decreases the stability (i.e., half-life) of BRM and / or BRG1. The polynucleotide may be chemically synthesized or transcribed in vitro.

[0111] Inhibitory polynucleotides can be designed by methods well known in the art, and siRNA, miRNA, shRNA, and asRNA molecules with sufficient homology to provide the sequence specificity required to uniquely degrade any RNA can be designed using programs known in the art, including but not limited to those maintained on the websites of Thermo Fisher Scientific, the German Cancer Research Center, and The Ohio State University Wexner Medical Center.Several types of systematic testing to optimize inhibitory polynucleotide sequences can be routinely performed by those skilled in the art.Considerations when designing interference polynucleotides include but are not limited to biophysical, thermodynamic, and structural considerations, base preferences at specific positions of the sense strand, and homology. In addition, the generation and use of inhibitory therapeutics based on non-coding RNA, such as ribozymes, RNase P, siRNA, and miRNA, are known in the art and are described, for example, in Sioud, RNA Therapeutics: Function, Design, and Delivery (Methods in Molecular Biology). Humana Press 2010.

[0112] The construction of vectors for expressing polynucleotides for use in the present invention can be accomplished using conventional techniques that do not require detailed explanation to those skilled in the art. The generation of efficient expression vectors requires the presence of control sequences that control the expression of the polynucleotide. These control sequences include promoter and enhancer sequences, are influenced by specific cellular factors that interact with these sequences, and are well known in the art.

[0113] Gene editing In some embodiments, the agent that reduces the level and / or activity of BRM and / or BRG1 is a component of a gene editing system. For example, the agent that reduces the level and / or activity of BRM and / or BRG1 introduces an alteration in BRM and / or BRG1 (e.g., an insertion, a deletion (e.g., a knockout), a transition, an inversion, a single point mutation, or other mutation). In some embodiments, the agent that reduces the level and / or activity of BRM and / or BRG1 is a nuclease. Exemplary gene editing systems include zinc finger nucleases (ZFNs), transcription activator-like effector-based nucleases (TALENs), and clustered regularly interspaced short palindromic repeats (CRISPR) systems. For ZFN, TALEN, and CRISPR-based methods, see, for example, Gaj et al., Trends Biotechnol. 31(7):397-405 (2013).

[0114] CRISPR refers to a set (or a system that includes a set) of clustered regularly interspaced short palindromic repeats. CRISPR system refers to a system derived from CRISPR and Cas (CRISPR associated proteins) or other nucleases that can be used to silence or mutate genes as described herein. CRISPR system is a naturally occurring system and is found in the genomes of bacteria and archaea. CRISPR loci are composed of alternating repeat sequences and spacer sequences. In naturally occurring CRISPR systems, the spacer is typically a sequence that is foreign to the bacteria (e.g., a plasmid or phage sequence). CRISPR system has been modified for use in gene editing (e.g., altering, silencing, and / or enhancing specific genes) in eukaryotes. See, for example, Wiedenheft et al., Nature 482(7385):331-338(2012). For example, such modification of the system includes introducing a specifically designed CRISPR and a plasmid containing one or more appropriate Cas proteins into a eukaryotic cell. CRISPR loci are transcribed into RNA and processed by Cas protein into small RNAs that contain repeat sequences flanked by spacers. The RNA acts as a guide to guide Cas protein to silence specific DNA / RNA sequences depending on the spacer sequence. See, for example, Horvath et al., Science 327(5962):167-170(2010); Makarova et al., Biology Direct 1:7(2006); Pennisi, Science 341(6148):833-836(2013). In some examples, CRISPR systems include Cas9 protein, a nuclease that cuts both strands of DNA. See, for example, ibid.

[0115] In some embodiments, in a CRISPR system for use described herein (e.g., for use with one or more methods described herein), the CRISPR spacer is derived from the sequence of the target gene (e.g., the sequence of BRM and / or BRG1).

[0116] In some embodiments, the agent that reduces the level and / or activity of BRM and / or BRG1 comprises a guide RNA (gRNA) for use in a CRISPR system for gene editing. Exemplary gRNAs for use in the methods of the present invention are provided in Table 1 below. In embodiments, the agent that reduces the level and / or activity of BRM and / or BRG1 comprises a ZFN that targets (e.g., cleaves) a nucleic acid sequence (e.g., a DNA sequence) of BRD7, or an mRNA encoding the ZFN. In embodiments, the agent that reduces the level and / or activity of BRM and / or BRG1 comprises a TALEN that targets (e.g., cleaves) a nucleic acid sequence (e.g., a DNA sequence) of BRM and / or BRG1, or an mRNA encoding the TALEN.

[0117] For example, gRNAs can be used in a CRISPR system to engineer modifications of genes (e.g., BRM and / or BRG1). In other examples, ZFNs and / or TALENs can be used to engineer modifications of genes (e.g., BRM and / or BRG1). Exemplary modifications include insertions, deletions (e.g., knockouts), translocations, inversions, single point mutations, or other mutations. Modifications can be introduced into genes in cells, for example, in vitro, ex vivo, or in vivo. In some embodiments, the modifications reduce (e.g., knock down or knock out) the level and / or activity of BRM and / or BRG1, e.g., the modifications are negative regulators of function. In yet another example, the modifications correct defects (e.g., mutations that cause defects) in BRM and / or BRG1.

[0118] In certain embodiments, the CRISPR system is used to edit the target gene, e.g., BRM and / or BRG1 (e.g., add or delete base pairs). In other embodiments, the CRISPR system is used to introduce a premature stop codon (e.g., thereby reducing expression of the target gene). In yet other embodiments, the CRISPR system is used to turn off the target gene in a reversible manner, e.g., similar to RNA interference. In embodiments, the CRISPR system is used to guide Cas to the promoter of the target gene, e.g., BRM and / or BRG1, thereby sterically blocking the RNA polymerase.

[0119] In some embodiments, a CRISPR system can be generated to edit BRM and / or BRG1 using techniques described, for example, in U.S. Publication No. 20140068797; Cong et al., Science 339(6121):819-823 (2013); Tsai, Nature Biotechnol., 32(6):569-576 (2014); and U.S. Patent Nos. 8,871,445, 8,865,406, 8,795,965, 8,771,945, and 8,697,359.

[0120] In some embodiments, CRISPR interference (CRISPRi) technology can be used for transcriptional repression of specific genes, such as genes encoding BRM and / or BRG1. In CRISPRi, an engineered Cas9 protein (e.g., nuclease null dCas9, or a dCas9 fusion protein, such as a dCas9-KRAB or dCas9-SID4X fusion) can pair with a sequence-specific guide RNA (sgRNA). The Cas9-gRNA complex can block RNA polymerase, thereby preventing transcription elongation. The complex can also block transcription initiation by preventing the binding of transcription factors. The CRISPRi method is specific, has minimal off-target effects, and can be multiplexed, e.g., to repress two or more genes simultaneously (e.g., using multiple gRNAs). The CRISPRi method also allows for reversible gene repression.

[0121] In some embodiments, CRISPR-mediated gene activation (CRISPRa) can be used for transcriptional activation of one or more genes, e.g., genes that inhibit BRM and / or BRG1, as described herein. In CRISPRa technology, dCas9 fusion proteins recruit transcriptional activators. For example, dCas9 can be used to recruit a polypeptide (e.g., an activation domain) such as VP64 or p65 activation domain (p65D) and used with sgRNA (e.g., a single sgRNA or multiple sgRNAs) to activate one or more genes, e.g., endogenous gene(s). Multiple activators can be recruited by using multiple sgRNAs, which can increase activation efficiency. Various activation domains and single or multiple activation domains can be used. In addition to engineering dCas9 to recruit activators, sgRNAs can also be engineered to recruit activators. For example, RNA aptamers can be incorporated into sgRNAs to recruit proteins (e.g., activation domains) such as VP64. In some instances, the synergistic activation mediator (SAM) system can be used for transcriptional activation. In SAM, the MS2 aptamer is added to the sgRNA. MS2 recruits the MS2 coat protein (MCP) fused to p65AD and heat shock factor 1 (HSF1). CRISPRi and CRISPRa technologies are described in more detail, for example, in Dominguez et al., Nat. Rev. Mol. Cell Biol. 17(1):5-15 (2016), incorporated herein by reference.

[0122] small molecule compound In some embodiments of the invention, the agent that reduces the level and / or activity of BRM and / or BRG1 in a cell is a small molecule compound. In some embodiments, the agent that reduces the level and / or activity of BRM and / or BRG1 has the following structure: [ka] has.

[0123] Other embodiments, and exemplary methods for the synthesis of the production of these compounds, are described herein.

[0124] Pharmaceutical Use The compounds described herein are useful in the methods of the invention and, without being bound by theory, are believed to exert their ability to modulate the level, status, and / or activity of the BAF complex, i.e., by inhibiting the activity of the BRG1 and / or BRM proteins within the mammalian BAF complex. BAF complex-associated disorders include, but are not limited to, disorders associated with loss-of-function mutations in BRG1.

[0125] One aspect of the invention relates to methods of treating a disorder associated with a loss-of-function mutation in BRG1, such as cancer (e.g., non-small cell lung cancer, colon cancer, bladder cancer, cancer of unknown primary site, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, or penile cancer) in a subject in need thereof. In some embodiments, the invention relates to methods of treating melanoma (e.g., uveal melanoma), prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or hematological cancer.

[0126] In some embodiments, the compound is administered in an amount and for a time that is effective to result in one or more (e.g., two or more, three or more, four or more) of: (a) a reduction in tumor size; (b) a reduction in tumor growth rate; (c) an increase in tumor cell death; (d) a reduction in tumor progression; (e) a reduction in the number of metastases; (f) a reduction in the rate of metastasis; (g) a reduction in tumor recurrence; (h) an increase in the survival rate of the subject; or (i) an increase in the progression-free survival of the subject.

[0127] Treating cancer can result in a reduction in tumor size or volume. For example, after treatment, tumor size is reduced by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) relative to its size before treatment. Tumor size can be measured by any reproducible means of measurement. For example, tumor size can be measured as the diameter of the tumor.

[0128] Cancer treatment can also result in a reduction in the number of tumors. For example, after treatment, the number of tumors is reduced by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) compared to the number before treatment. The number of tumors can be measured by any reproducible measurement means, for example, the number of tumors can be measured by counting the tumors that are visible to the naked eye or at a certain magnification (e.g., 2x, 3x, 4x, 5x, 10x, or 50x).

[0129] Treatment of cancer can result in a reduction in the number of metastatic nodules in other tissues or organs distant from the primary tumor site. For example, after treatment, the number of metastatic nodules is reduced by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) compared to the number before treatment. The number of metastatic nodules can be measured by any reproducible measurement means. For example, the number of metastatic nodules can be measured by counting the metastatic nodules that are visible to the naked eye or at a certain magnification (e.g., 2x, 10x, or 50x).

[0130] Treating cancer can result in an increase in the average survival time of a population of subjects treated according to the present invention compared to a population of untreated subjects. For example, the average survival time increases by more than 30 days (more than 60 days, 90 days, or 120 days). The increase in the average survival time of a population can be measured by any reproducible means. The increase in the average survival time of a population can be measured, for example, by calculating the average survival time of a population after the start of treatment with a compound of the present invention. The increase in the average survival time of a population can also be measured, for example, by calculating the average survival time of a population after the completion of the first round of treatment with a pharma- ceutically acceptable salt of a compound of the present invention.

[0131] Treating cancer may also result in a reduction in the mortality rate of a population of treated subjects compared to an untreated population. For example, the mortality rate is reduced by more than 2% (for example, more than 5%, 10%, or 25%). The reduction in the mortality rate of a population of treated subjects may be measured by any reproducible means, for example, by calculating the average number of disease-related deaths per unit time for a population after the start of treatment with a pharmaceutically acceptable salt of the present invention. The reduction in the mortality rate of a population may also be measured, for example, by calculating the average number of disease-related deaths per unit time for a population after the completion of the first round of treatment with a pharmaceutically acceptable salt of the present invention.

[0132] Exemplary cancers that may be treated by the present invention include, but are not limited to, non-small cell lung cancer, small cell lung cancer, colon cancer, bladder cancer, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, esophagogastric cancer, esophageal cancer, pancreatic cancer, hepatobiliary cancer, soft tissue sarcoma, ovarian cancer, head and neck cancer, renal cell carcinoma, bone cancer, non-Hodgkin's lymphoma, prostate cancer, embryonal tumors, germ cell tumors, cervical cancer, thyroid cancer, salivary gland cancer, gastrointestinal neuroendocrine tumors, uterine sarcoma, gastrointestinal stromal tumors, CNS cancer, thymic tumor, adrenocortical carcinoma, appendix cancer, small intestine cancer, blood cancer, and penile cancer.

[0133] Combination preparations and their uses The compounds of the invention can be combined with one or more therapeutic agents. In particular, the therapeutic agents can be those that treat or prophylactically treat any of the cancers described herein.

[0134] Combination therapy The compounds of the present invention can be used alone or in combination with additional therapeutic agents, such as other drugs that treat cancer or related symptoms, or in combination with other types of treatments to treat cancer. In combination treatment, the dosage of one or more therapeutic compounds can be reduced from the standard dosage when administered alone. For example, dosage can be empirically determined from drug combinations and permutations, or estimated by isobolographic analysis (e.g., Black et al., Neurology 65:S3-S6, 2005). In this case, the dosage of the compounds when combined should provide therapeutic effect.

[0135] In some embodiments, the second therapeutic agent is a chemotherapeutic agent (e.g., a cytotoxic agent or other compound useful in the treatment of cancer). These include alkylating agents, antimetabolites, folic acid analogs, pyrimidine analogs, purine analogs and related inhibitors, vinca alkaloids, epipodophyllotoxins, antibiotics, L-asparaginase, topoisomerase inhibitors, interferons, platinum coordination complexes, anthracenedione-substituted ureas, methylhydrazine derivatives, adrenocorticotropic inhibitors, adrenocorticosteroids, progestins, estrogens, antiestrogens, androgens, antiandrogens, and gonadotropin-releasing hormone analogs. Also included are 5-fluorouracil (5-FU), leucovorin (LV), irenotecan, oxaliplatin, capecitabine, paclitaxel, and doxetaxel. Non-limiting examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide, alkyl sulfonates such as busulfan, improsulfan, and piposulfan, aziridines such as benzodopa, carboquone, meturedopa, and uredopa, ethylenimines and methylameramines such as altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine, acetogenins (specifically bullatacin and bullatacinone), camptothecins (including the synthetic analog topotecan), bryostatin, kallistatin, CC-1065 (including its synthetic analogs adozelesin, carzelesin, and bezelesin), chlosporins, cyclosporins, and cyclosporins. Cryptophycins (specifically cryptophycin 1 and cryptophycin 8), dolastatins, duocarmycins (including the synthetic analogs KW-2189 and CB1-TM1), erytherobin, pancratistatin, sarcodictyin, spongistatin, nitrogen mustards such as chlorambucil, chlornaphazine, colofosfamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nobembitine, phenesterine, prednimustine, trofosfamide, uracil mustard, nitrosoureas such as camrustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine, antibiotics such asEnediyne antibiotics (e.g., the calicheamicins, specifically calicheamicin gamma II and calicheamicin omega II (see, e.g., Agnew, Chem. Intl. Ed Engl. 33:183-186 (1994)); the dynemicins, including dynemicin A; bisphosphonates, such as clodronate; esperamicin; and neocarzinostatin chromophore and related enediyne antibiotic chromophores), aclacinomycin, actinomycin, authramycin, azaserine, bleomycin, cactinomycin, carabimycin, caminomycin, caminomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, Adriamycin® (doxorubicin, including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcelomycin, myocin, Mitomycins such as tomycin C, mycophenolic acid, nogalamycin, olivomycin, peplomycin, potfilomycin, puromycin, keramycin, rhodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin, methotrexate, and 5-fluorouracil (5-FU); antimetabolites such as denopterin, methotrexate, pteropterin, trimetrexate, and the like. folic acid analogues;purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine;pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, and floxuridine;androgens such as calsterone, dromostanolone propionate, epithiostanol, mepitiostane, and testolactone;aminoglutethimide, mitotane,Anti-adrenal agents such as trilostane; folic acid replacement fluids such as floric acid; aceglatone; aldophosphamide glycosides; aminolevulinic acid; eniluracil; amsacrine; bestravcil; bisantrene; edatrexate; defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate; epothilone; etoglucide; gallium nitrate; hydroxyurea; lentinan; maytansinoids such as lonidynin, maytansine and ansamitocin; mitoguazone, mitoxantrone, mopidanmol, nitraelin, pentostatin; fenameth; pirarubicin; rosoxantrone; podophyllic acid; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, Oreg.), razoxane; rhizoxin; schizofiran; spirogermanium; tenuazonic acid; triazicon; 2,2',2''-trichlorotriethylamine; trichothecenes (especially T-2 toxin, veraculin A, roridin A, and anguidine); urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids such as Taxol® paclitaxel (Bristol-Myers Squibb Oncology, Princeton, NJ), ABraxane®, a chromophore-free, albumin engineered nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.), and Taxotere® doxetaxel (Rhone-Poulenc Rorer, Antony, France); chlorambucil; Gemzar® gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; cisplatin, oxaliplatin,and platinum coordination complexes such as carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; Navelbine® vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; irinotecan (e.g., CPT-11); topoisomerase inhibitors RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; capecitabine; and pharma- ceutical acceptable salts, acids, or derivatives of any of the above. Two or more chemotherapeutic agents can be used in a cocktail administered in combination with a first therapeutic agent described herein. Suitable dosing regimens for combination chemotherapy are known in the art and are described, for example, in Saltz et al. (1999) Proc ASCO 18:233a and Douillard et al. (2000) Lancet 355:1041-7.

[0136] In some embodiments, the second therapeutic agent is a therapeutic agent that is a biologic, such as a cytokine (e.g., an interferon or an interleukin (e.g., IL-2)) used in cancer treatment. In some embodiments, the biologic is an anti-VEGF agent, e.g., an anti-angiogenic agent, such as bevacizumab (Avastin®). In some embodiments, the biologic is an immunoglobulin-based biologic, e.g., a monoclonal antibody (e.g., a humanized antibody, a fully human antibody, an Fc fusion protein, or a functional fragment thereof) that agonizes a target to stimulate an anti-cancer response or antagonizes an antigen important to cancer. Such medications include Rituxan (rituximab), Zenapax (daclizumab), Simulect (basiliximab), Synagis (palivizumab), Remicade (infliximab), Herceptin (trastuzumab), Mylotarg (gemtuzumab ozogamicin), Campath (alemtuzumab), Zevalin (ibritumomab tiuxetan), Humira (adalimumab), Xolair (omalizumab), Bexxar (tositumomab-I-131), Raptiva (efalizumab), Erbitux (cetuximab), Avastin (bevacizumab), Tysabri (natalizumab), Actemra (tosi Antibody-drug conjugates are also included.

[0137] The second agent can be a therapeutic agent that is a non-drug treatment, for example, the second therapeutic agent is radiation therapy, cryotherapy, thermotherapy, and / or surgical removal of tumor tissue.

[0138] The second agent may be a checkpoint inhibitor. In one embodiment, the checkpoint inhibitor is an inhibitory antibody (e.g., a monospecific antibody such as a monoclonal antibody). The antibody may be, for example, humanized or fully human. In some embodiments, the checkpoint inhibitor is a fusion protein, e.g., an Fc receptor fusion protein. In some embodiments, the checkpoint inhibitor is an agent, such as an antibody, that interacts with a checkpoint protein. In some embodiments, the checkpoint inhibitor is an agent, such as an antibody, that interacts with a ligand of the checkpoint protein. In some embodiments, the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of CTLA-4 (e.g., an anti-CTLA4 antibody such as ipilimumab / Yervoy or tremelimumab). In some embodiments, the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of PD-1 (e.g., nivolumab / Opdivo®; pembrolizumab / Keytruda®; pidilizumab / CT-011). In some embodiments, the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of PD-L1 (e.g., MPDL3280A / RG7446; MEDI4736; MSB0010718C; BMS 936559). In some embodiments, the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibody or Fc fusion or small molecule inhibitor) of PDL2 (e.g., a PDL2 / Ig fusion protein such as AMP 224). In some embodiments, the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of B7-H3 (e.g., MGA271), B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, B-7 family ligand, or combinations thereof.

[0139] In some embodiments, the compounds of the present invention are used in combination with another anti-cancer therapy used to treat uveal melanoma, such as surgery, MEK inhibitors, and / or PKC inhibitors.For example, in some embodiments, the method further comprises performing surgery before, after, or simultaneously with administration of the compounds of the present invention.In some embodiments, the method further comprises administering a MEK inhibitor (e.g., selumetinib, binimetinib, or tametinib) and / or a PKC inhibitor (e.g., sotrastaurin or IDE196) before, after, or simultaneously with administration of the compounds of the present invention.

[0140] In any of the combination embodiments described herein, the first and second therapeutic agents are administered simultaneously or sequentially, in either order. The first therapeutic agent may be administered immediately before, immediately after, up to 1 hour, up to 2 hours, up to 3 hours, up to 4 hours, up to 5 hours, up to 6 hours, up to 7 hours, up to 8 hours, up to 9 hours, up to 10 hours, up to 11 hours, up to 12 hours, up to 13 hours, 14 hours, up to 16 hours, up to 17 hours, up to 18 hours, up to 19 hours, up to 20 hours, up to 21 hours, up to 22 hours, up to 23 hours, up to 24 hours, or up to 1-7, 1-14, 1-21, or 1-30 days before or after the second therapeutic agent.

[0141] Pharmaceutical Compositions The compounds of the present invention are preferably formulated into pharmaceutical compositions for administration to mammals, preferably humans, in a biologically compatible form suitable for administration in vivo. Thus, in one aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention mixed with a suitable diluent, carrier, or excipient.

[0142] The compounds of the present invention may be used in the form of free base, salt, solvate, and prodrug. All forms are within the scope of the present invention. According to the method of the present invention, as understood by those skilled in the art, the described compounds, or their salts, solvates, or prodrugs, may be administered to patients in various forms depending on the selected route of administration. The compounds of the present invention may be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump, or transdermal administration, and the pharmaceutical composition is formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal, and topical modes of administration. Parenteral administration may be by continuous infusion over a selected period of time.

[0143] The compounds of the present invention can be administered orally, for example, with an inert diluent or an assimilable edible carrier, or can be enclosed in hard or soft shell gelatin capsules, or can be compressed into tablets, or can be incorporated directly with the food of the diet. For oral therapeutic administration, the compounds of the present invention can be incorporated with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, and wafers.

[0144] The compounds of the present invention may also be administered parenterally. Solutions of the compounds of the present invention may be prepared in water suitably mixed with a surfactant such as hydroxypropylcellulose. Dispersions may also be prepared in glycerol, liquid polyethylene glycols, DMSO, and mixtures thereof with or without alcohol, as well as in oils. These preparations may contain a preservative to prevent the growth of microorganisms under ordinary storage and use conditions. Conventional procedures and ingredients for the selection and preparation of suitable formulations are described, for example, in Remington's Pharmaceutical Sciences (2003, 20th ed.) and The United States Pharmacopeia: The National Formulary, published in 1999 (USP 24 NF19). Pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be sufficiently fluid to be easily administered via syringe.

[0145] The compounds described herein may be administered intratumorally, for example, as intratumoral injection. Intratumoral injection is a direct injection into tumor vasculature, and is particularly contemplated for individual solid accessible tumors. Local, regional, or systemic administration may also be appropriate. The compounds described herein may be advantageously contacted by administering an injection or multiple injections to the tumor, for example, spaced approximately 1 cm apart. In the case of surgical intervention, the present invention may be used prior to surgery, such as to subject inoperable tumors to resection. Continuous administration may also be applied where appropriate, for example, by implanting a catheter into the tumor or tumor vasculature.

[0146] The compounds of the invention, as described herein, may be administered to animals, e.g., humans, alone or in combination with pharma- ceutically acceptable carriers, the ratio of which will be determined by the solubility and chemical nature of the compound, the chosen route of administration, and standard pharmaceutical practice.

[0147] Dosage The dosage of the compound of the present invention and / or the composition containing the compound of the present invention may vary depending on many factors, such as the pharmacodynamic properties of the compound; the mode of administration; the age, health, and weight of the recipient; the nature and extent of the symptoms; the frequency of treatment and the type of concomitant treatment, if any; and the clearance rate of the compound in the treated animal. Those skilled in the art can determine the appropriate dosage based on the above factors. The compound of the present invention may be administered at a suitable dosage initially, and this amount may be adjusted as necessary depending on the clinical response. In general, satisfactory results can be obtained when the compound of the present invention is administered to humans at a daily dose of, for example, 0.05 mg to 3000 mg (measured as solid form).

[0148] Alternatively, the patient's body weight can be used to calculate the dosage. For example, the dose of the compound or pharmaceutical composition thereof administered to a patient may range from 0.1 to 50 mg / kg.

[0149] In some embodiments, the dose of the compound, drug, or pharmaceutical composition thereof (e.g., Compound 1) may be 1 mg to 15 mg (e.g., about 1 mg to 2.5 mg, about 2.5 mg to 5 mg, about 5 mg to 7.5 mg, or about 7.5 mg to about 10 mg). In some embodiments, the dose of the compound, drug, or pharmaceutical composition thereof (e.g., Compound 1) is about 2 mg to 3 mg (e.g., about 2.5 mg). In some embodiments, the dose of the compound, drug, or pharmaceutical composition thereof (e.g., Compound 1) is about 4 mg to 6 mg (e.g., about 5 mg). In some embodiments, the dose of the compound, drug, or pharmaceutical composition thereof (e.g., Compound 1) is about 7 mg to 8 mg (e.g., about 7.5 mg). In some embodiments, the dose of the compound, drug, or pharmaceutical composition thereof (e.g., Compound 1) is about 9 mg to 11 mg (e.g., about 10 mg).

[0150] In some embodiments, the compound, agent, or pharmaceutical composition thereof (e.g., Compound 1) is administered in two or more doses, the doses being administered once daily, twice daily (BID), once weekly, once every two weeks, or once monthly. In some embodiments, administration includes multiple doses comprising a period of at least 7 days, e.g., at least 7 days, 8 days, 9 days, 10 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, or more.

[0151] In some embodiments, the compound, agent, or pharmaceutical composition thereof (e.g., Compound 1) is administered once daily for a week with one week of rest for one or more cycles. In some embodiments, the compound, agent, or pharmaceutical composition thereof (e.g., Compound 1) is administered once daily for a week with one week of rest for one or more cycles at a dose described herein (e.g., a dose of about 2.5 mg, 5 mg, 7.5 mg, or 10 mg).

[0152] In some embodiments, the compound, agent, or pharmaceutical composition thereof (e.g., Compound 1) is administered once daily for two weeks with one week of rest for one or more cycles. In some embodiments, the compound, agent, or pharmaceutical composition thereof (e.g., Compound 1) is administered once daily for two weeks with one week of rest for one or more cycles at a dose described herein (e.g., a dose of about 2.5 mg, 5 mg, 7.5 mg, or 10 mg). EXAMPLES

[0153] The following abbreviations are used throughout the Examples section: [Table 1]

[0154] Example 1. Preparation of N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide was synthesized as shown in Scheme 1 below. [ka]

[0155] Step 1: Preparation of 6-fluoropyridine-2-carbonyl chloride (Intermediate B) [ka] To a cooled (0° C.) solution of 6-fluoropyridine-2-carboxylic acid (50.0 g, 354 mmol) in dichloromethane (500 mL) and N,N-dimethylformamide (0.26 mL, 3.54 mmol) was added oxalyl chloride (155 mL, 1.77 mol). After complete addition of oxalyl chloride, the reaction mixture was allowed to warm to room temperature. After 0.5 h, the mixture was concentrated in vacuo to give intermediate B (56.50 g) as a white solid, which was used in the next step without further purification.

[0156] Step 2: Preparation of 2-chloro-1-(6-fluoro-2-pyridyl)ethenone (Intermediate C) [ka] To a cooled (0° C.) mixture of intermediate B (56.0 g, 351 mmol) in 1,4-dioxane (800 mL) was added dropwise a solution of 2 M trimethylsilyldiazomethane in hexanes (351 mL, 702 mmol). The resulting reaction mixture was stirred at 25° C. for 10 h. The reaction mixture was subsequently quenched with a solution of 4 M HCl in 1,4-dioxane (500 mL, 2.0 mol). After stirring for 2 h, the reaction solution was concentrated under vacuum to give an oil. The residue was purified by distillation with NaHCO 3 The mixture was diluted with saturated aqueous solution and extracted three times with ethyl acetate. The combined organic layers were washed with brine and 2 SO 4 Drying at 40° C., filtering and concentrating under reduced pressure gave Intermediate C (35.5 g) as a white solid which was used directly in the next step. LCMS(ESI)m / z:[M+H] + =173.8.

[0157] Step 3: Preparation of 4-(6-fluoro-2-pyridyl)thiazol-2-amine (Intermediate E) [ka] To a solution of intermediate C (35.5 g, 205 mmol) and thiourea (14.0 g, 184 mmol) in a mixture of methanol (250 mL) and water (250 mL) was added NaF (3.56 g, 84.8 mmol) at room temperature. After stirring for 0.5 h, the reaction mixture was partially concentrated in vacuo to remove MeOH and the resulting solution was acidified to pH ∼3 with 2 M aqueous HCl. After 15 min, the solution was extracted three times with ethyl acetate. The organic layer was discarded and the aqueous phase was washed with NaHCO 3 The mixture was alkalized with saturated aqueous solution, stirred for 30 min, and extracted three times with ethyl acetate. The combined organic layers were washed three times with brine and 2 SO 4 The mixture was dried at 40° C., filtered and concentrated under reduced pressure. The residue was triturated with petroleum ether, stirred at 25° C. for 10 min and filtered. The resulting solid was dried under vacuum to give Intermediate E (28.0 g, 143 mmol, 70.1% yield, 100% purity) as a white solid. LCMS(ESI)m / z:[M+H] += 195.8. 1 H NMR (400MHz, DMSO-d 6 )δ 8.00-7.96(m,1H),7.72(d,J=7.2Hz,1H),7.24(s,1H),7.16(s,2H),7.02(d,J=8.0Hz,1H).

[0158] Step 4: Preparation of 4-[6-[cis-2,6-dimethylmorpholin-4-yl]-2-pyridyl]thiazol-2-amine (Intermediate G) [ka] Ten separate mixtures of intermediate E (2.00 g, 10.3 mmol), cis-2,6-dimethylmorpholine (3.54 g, 30.7 mmol), and DIPEA (5.35 mL, 30.7 mmol) in dimethyl sulfoxide (10 mL) were 2 The mixture was stirred in parallel under atmosphere at 120° C. After 36 h, the reaction mixtures were combined and added dropwise to water. The resulting suspension was filtered, and the filter cake was washed three times with water and once with petroleum ether, then dried under reduced pressure to give intermediate G (25.5 g, 87.8 mmol, 95.2% yield) as a yellow solid. LCMS(ESI)m / z:[M+H] + =291.2. 1 H NMR (400MHz, DMSO-d 6 )δ 7.56-7.54(m,1H),7.17(s,1H),7.13(d,J=7.6Hz,1H),7.01(s,2H),6.72(d,J=8.8Hz ,1H),4.26-4.15(m,2H),3.67-3.55(m,2H),2.38-2.34(m,2H),1.17(d,J=6.4Hz,6H).

[0159] Step 5: Preparation of tert-butyl N-[(1S)-2-[[4-[6-[cis-2,6-dimethylmorpholin-4-yl]-2-pyridyl]thiazol-2-yl]amino]-1-(methoxymethyl)-2-oxo-ethyl]carbamate (Intermediate I) [ka] To a solution of intermediate G (12.0 g, 41.3 mmol) and (2S)-2-(tertbutoxycarbonylamino)-3-methoxy-propanoic acid (10.9 g, 49.6 mmol) in dichloromethane (60 mL) was added EEDQ (12.3 g, 49.6 mmol). After stirring at room temperature for 16 h, the reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=2:1-3:2) to give intermediate I (20.0 g, 40.7 mmol, 98.5% yield) as a yellow viscous material. LCMS(ESI)m / z:[M+H] + =492.2. 1 H NMR (400MHz, DMSO-d 6 )δ 12.37(s,1H),7.78(s,1H),7.64-7.60(m,1H),7.25(d,J=7.2Hz,1H),7.16(d,J=7.2Hz,1H),6.79(d,J=8.4Hz,1H),4.50-4 .48(m,1H),4.25(d,J=11.6Hz,2H),3.70-3.51(m,4H),3.26(s,3H),2.44-2.40(m,2H),1.39(s,9H),1.18(d,J=6.4Hz,6H).

[0160] Step 6: Preparation of (S)-4-(4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)-1-methoxy-3-oxobutan-2-aminium chloride (Intermediate J) [ka] To a solution of 4M HCl in 1,4-dioxane (200 mL, 800 mmol) was added a solution of intermediate I (20.0 g, 40.7 mmol) in dichloromethane (50 mL). After stirring at room temperature for 2 h, the mixture was diluted with methyl tert-butyl ether to give a suspension. The solid was collected by filtration, washed twice with methyl tert-butyl ether, and dried in vacuum to give intermediate J (19.0 g) as a yellow solid, which was used in the next step without further purification. LCMS(ESI)m / z:[M+H] + =392.3. 1 H NMR (400MHz, DMSO-d 6 )δ 13.44-12.30(m,1H),8.65(d,J=4.4Hz,3H),7.87(s,1H),7.66-7.64(m,1H),7.25(d,J=7.2Hz,1H),6.83(d,J=8.8Hz,1H),4.39-4.30(m ,1H),4.25(d,J=11.6Hz,2H),3.94-3.86(m,1H),3.85-3.77(m,1H),3.69-3.57(m,2H),3.31(s,3H),2.43(m,2H),1.18(d,J=6.4Hz,6H).

[0161] Preparation of 1-(methylsulfonyl)-1H-pyrrole-3-carboxylic acid (intermediate K) 1-(Methylsulfonyl)-1H-pyrrole-3-carboxylic acid was synthesized as shown in Scheme 2 below. [ka]

[0162] Step A: Preparation of tert-butyl 1H-pyrrole-3-carboxylate (Intermediate N) [ka] To a mixture of tert-butyl-prop-2-enoate (78.6 mL, 542 mmol) and 1-(isocyanomethylsulfonyl)-4-methylbenzene (106 g, 542 mmol) in THF (1300 mL) was added 60% NaH in mineral oil (25.97 g, 649 mmol) slowly over 1 h at 30 °C and then heated to 70 °C. After 2 h, the reaction mixture was diluted with saturated NH 4 The combined organic phase was washed twice with brine and extracted with anhydrous NaCl. 2 SO 4 The mixture was dried at 40° C., filtered, and concentrated under reduced pressure to give a residue which was purified by silica gel column chromatography (petroleum ether:ethyl acetate=20:1 to 3:1) to give intermediate N (41.5 g, 236 mmol, 43% yield) as a yellow solid. LCMS(ESI)m / z[M+Na] + =180.4. 1 H NMR (400 MHz, CDCl 3 )δ 8.36(br s,1H),7.35-7.25(m,1H),6.71-6.62(m,1H),6.59-6.49(m,1H),1.48(s,9H).

[0163] Step B: Preparation of tert-butyl 1-methylsulfonylpyrrole-3-carboxylate (Intermediate O) [ka] To a cooled (0° C.) solution of intermediate N (40.5 g, 242 mmol) in THF (1500 mL) was added a 1 M solution of NaHMDS (484 mL, 484 mmol). After stirring at 0° C. for 30 min, methanesulfonyl chloride (28.1 mL, 363 mmol) was added slowly and the mixture was warmed to 30° C. After 16 h, the reaction mixture was diluted with saturated NH 4 The combined organic layer was washed twice with brine and then with anhydrous NaCl. 2 SO 4The mixture was dried at 40° C., filtered, and concentrated under reduced pressure to give a residue. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate=10:1) to give a yellow solid. The yellow solid was triturated with methyl tert-butyl ether at room temperature, stirred for 20 minutes, filtered, and dried in vacuum to give intermediate O (25.7 g, 105 mmol, 43% yield) as a white solid. 1 H NMR (400 MHz, CDCl 3 )δ 7.66-7.64(m,1H),7.10-7.08(m,1H),6.73-6.71(m,1H),3.21(s,3H),1.56(s,9H).

[0164] Step C: Preparation of 1-methylsulfonylpyrrole-3-carboxylic acid (Intermediate K) [ka] To a mixture of intermediate O (25.7 g, 105 mmol) in 1,4-dioxane (100 mL) was added a 4 M solution of HCl in 1,4-dioxane (400 mL, 1.6 mol) at 15° C. After stirring at 15° C. for 14 h, the reaction mixture was concentrated under reduced pressure to give a residue. The residue was triturated with methyl tert-butyl ether at 15° C. for 16 h. The mixture was filtered and dried in vacuum to give intermediate K (18.7 g, 98.8 mmol, 94% yield) as a white solid. LCMS(ESI)m / z[M+H] + =189.8. 1 H NMR (400MHz, methanol-d 4 )δ 7.78-7.77(m,1H),7.25-7.23(m,1H),6.72-6.70(m,1H),3.37(s,3H).

[0165] Step 7: Preparation of N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide [ka] To a solution of 1-methylsulfonylpyrrole-3-carboxylic acid (Intermediate K) (2.43 g, 12.9 mmol), EDCI (2.69 g, 14.0 mmol), HOBt (1.89 g, 14.0 mmol), and DIPEA (10.2 mL, 58.4 mmol) in dichloromethane (50 mL) was added Intermediate J (5.00 g, 11.7 mmol). After stirring at room temperature for 4 h, the reaction mixture was concentrated under reduced pressure. The residue was diluted with water and extracted three times with ethyl acetate. The combined organic layers were washed with saturated NH 4 Wash three times with aqueous Cl, once with brine, and 2 SO 4 The mixture was dried at 40° C., filtered, and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=1:1 to 1:2). The residue was triturated with methyl tert-butyl ether. After 0.5 h, the suspension was filtered, and the filter cake was washed with methyl tert-butyl ether and dried in vacuum. The solid was dissolved in dimethyl sulfoxide (12 mL) and added dropwise to water (800 mL). The suspension was filtered to give a wet filter cake. The filter cake was suspended in water and stirred at room temperature. After 1 h, the solid was collected by filtration, washed three times with water and dried under vacuum to give N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide (3.9 g, 6.93 mmol, 59.3% yield) as a white solid. LCMS(ESI)m / z:[M+H] + =563.1. 1H NMR (400MHz, DMSO-d 6 )δ 12.49(br s,1H),8.51(d,J=7.2Hz,1H),7.98-7.97(m,1H),7.78(s,1H),7.67-7.57 (m,1H),7.29-7.27(m,1H),7.26(d,J=7.2Hz,1H),6.88-6.74(m,2H),4.9 4-4.91(m,1H),4.25(d,J=11.6Hz,2H),3.77-3.67(m,2H),3.63-3.62(m, 2H),3.57(s,3H),3.31(s,3H),2.44-2.38(m,2H),1.18(d,J=6.0Hz,6H).

[0166] Example 2. N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-(methoxy-d 3 )-1-Oxopropan-2-yl-3,3-d 2 Preparation of 1-(methylsulfonyl)-1H-pyrrole-3-carboxamide [ka] N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-(methoxy-d 3 )-1-Oxopropan-2-yl-3,3-d 2 )-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide was prepared according to the synthesis protocol described in Example 1, and intermediate H was converted to N-(tert-butoxycarbonyl)-O-(methyl-d 3 )-L-Serine-3,3-d 2 Replaced with N-(tert-butoxycarbonyl)-O-(methyl-d 3 )-L-Serine-3,3-d 2 was prepared from isotopically enriched material according to the synthetic procedure described by A. Yang et al., Org. Process Res. Dev. 2019, 23, 818-824. LCMS(ESI)m / z:[M+H]+ =568.2. 1 H NMR (400MHz, DMSO-d 6 )δ 12.45(s,1H),8.47(d,J=7.2Hz,1H),7.98(dd,J=2.3,1.7Hz,1H),7.78(s,1H), 7.62(dd,J=8.5,7.4Hz,1H),7.29(dd,J=3.2,2.3Hz,1H),7.26(d,J=7.3Hz,1H) ,6.84-6.75(m,2H),4.91(d,J=7.2Hz,1H),4.25(dd,J=13.1,2.3Hz,2H),3.69- 3.59(m,2H),3.56(s,3H),2.42(dd,J=12.8,10.5Hz,2H),1.18(d,J=6.2Hz,6H).

[0167] Example 3. Preparation of N-((R)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-(methoxy)-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide N-((R)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-(methoxy)-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide was prepared according to the synthetic protocol described in Example 1, replacing intermediate H with (2R)-2-(tertbutoxycarbonylamino)-3-methoxy-propanoic acid. LCMS(ESI)m / z:[M+H] + =563.1. 1 H NMR (400MHz, DMSO-d 6)δ 12.5(s,1H),8.50(d,J=7.2Hz,1H),7.98(t,J=1.6Hz,1H),7.78(s,1H),7.62(dd,J=7 .2,8.4Hz,1H),7.29(dd,J=2.0,3.2Hz,1H),7.26(d,J=7.2Hz,1H),6.79-6.81(m,2H), 4.92(q,J=6.4,12.8Hz,1H),4.25(d,J=11.2Hz,2H),3.69-3.75(m,2H),3.59-3.66(m ,2H),3.56(s,3H),3.31(s,3H),2.41(dd,J=10.8,12.8Hz,2H),1.18(d,J=6.0Hz,6H).

[0168] Example 4. N-((R)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-(methoxy-d 3 )-1-Oxopropan-2-yl-3,3-d 2 Preparation of 1-(methylsulfonyl)-1H-pyrrole-3-carboxamide N-((R)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-(methoxy-d 3 )-1-Oxopropan-2-yl-3,3-d 2 )-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide was prepared according to the synthesis protocol described in Example 1, and intermediate H was converted to N-(tert-butoxycarbonyl)-O-(methyl-d 3 )-D-Serine-3,3-d 2 Replaced with N-(tert-butoxycarbonyl)-O-(methyl-d 3 )-D-Serine-3,3-d 2 was prepared from isotopically enriched material according to the synthetic procedure described by A. Yang et al., Org. Process Res. Dev. 2019, 23, 818-824. LCMS(ESI)m / z:[M+H] + =568.3. 1 H NMR (400MHz, DMSO-d6 ) δ 12.46(s,1H),8.52-8.38(m,1H),7.97(t,J=1.9Hz,1H),7.76(s,1H),7.62(dd,J =8.5,7.3Hz,1H),7.29(dd,J=3.3,2.3Hz,1H),7.26(d,J=7.4Hz,1H),6.79(dt,J= 5.1,1.8Hz,2H),4.89(d,J=5.2Hz,1H),4.31-4.20(m,2H),3.63(ddd,J=10.5,6. 2,2.5Hz,2H),3.56(s,3H),2.41(dd,J=12.8,10.5Hz,2H),1.18(d,J=6.2Hz,6H).

[0169] Example 5. Assay of ATPase catalytic activity of BRM and BRG-1 The ATPase catalytic activity of BRM or BRG-1 was measured by an in vitro biochemical assay using ADP-Glo™ (Promega, V9102). Once the reaction was completed, the ADP-Glo™ kinase assay was performed in two steps. The first step was to deplete any ATP that was not consumed during the reaction. The second step was to convert the reaction product, ADP, to ATP, which was utilized to generate luminescence by luciferase and detected by a luminescence reader such as Envision.

[0170] The assay reaction mixture (10 μL) contained 30 nM BRM or BRG-1, 20 nM salmon sperm DNA (Invitrogen, UltraPure™ Salmon Sperm DNA Solution, Catalog No. 15632011), and 400 μM ATP in ATPase assay buffer, which is 20 mM Tris (pH 8), 20 mM MgCl . 2, 50 mM NaCl, 0.1% Tween-20, and 1 mM fresh DTT (Pierce™ DTT (dithiothreitol), catalog number 20290). The reaction was started by adding 2.5 μL of ATPase solution to 2.5 μL of ATP / DNA solution on a small volume white Proxiplate-384 plus plate (PerkinElmer, catalog number 6008280) and incubated for 1 hour at room temperature. The reaction was then incubated for 40 minutes at room temperature after adding 5 μL of ADP-Glo™ reagent provided in the kit. 10 μL of Kinase Detection Reagent provided in the kit was then added to convert ADP to ATP, and the reaction was incubated for 60 minutes at room temperature. Finally, luminescence measurements are collected with a plate-reading luminometer such as the Envision.

[0171] BRM and BRG-1 were synthesized from the High Five insect cell line with a purity of greater than 90%.

[0172] N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide had an IP of 3.9 nM for BRM and 5.2 nM for BRG1 in the assay. 50 N-((R)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-(methoxy-d3)-1-oxopropan-2-yl-3,3-d2)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide had an IP of 443 nM for BRM and 777 nM for BRG1 in the assay. 50 It was found to have the formula: N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-d 3 )-1-Oxopropan-2-yl-3,3-d 2)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide had an IP of 4.6 nM for BRM and 7.4 nM for BRG1 in the assay. 50 It was found to have the following structure:

[0173] Example 6. Synthesis of Compound A BRG1 / BRM inhibitor compound A has the following structure: [ka]

[0174] Compound A was synthesized as shown in Scheme 3 below. [ka]

[0175] The ATPase catalytic activity of BRM or BRG-1 in the presence of Compound A was measured by an in vitro biochemical assay using ADP-Glo™ (Promega, V9102) as described above. Compound A had an IP of 10.4 nM for BRM and 19.3 nM for BRG1 in the assay. 50 It was found to have the following structure:

[0176] Example 7. Effect of BRG1 / BRM ATPase inhibition on the proliferation of uveal melanoma and blood cancer cell lines Procedure: Uveal melanoma cell lines (92-1, MP41, MP38, MP46), prostate cancer cell line (LNCAP), lung cancer cell line (NCI-H1299), and immortalized embryonic kidney line (HEK293T) were seeded into 96-well plates containing growth medium (see Table 1). Compound A, a BRG1 / BRM ATPase inhibitor, was dissolved in DMSO and added to the cells at the time of seeding in a concentration gradient of 0-10 μM. Cells were incubated at 37°C for 3 days. After 3 days of treatment, the medium was removed from the cells and 30 microliters of TrypLE (Gibco) was added to the cells for 10 minutes. Cells were detached from the plate and resuspended by adding 170 microliters of growth medium. From the two DMSO-treated control wells, cells were counted and the initial cell numbers plated at the beginning of the experiment were re-plated into fresh compound-containing plates for an additional 4 days at 37°C. On day 7, cells were harvested as described above. On days 3 and 7, relative cell proliferation was measured by adding Cell-titer glo (Promega) and measuring luminescence with an Envision plate reader (Perkin Elmer). The concentration of compound at which proliferation of each cell line was inhibited by 50% (GI 50) was calculated using Graphpad Prism and plotted below. The above method was performed for multiple myeloma cell lines (OPM2, MM1S, LP1), ALL cell lines (TALL1, JURKAT, RS411), DLBCL cell lines (SUDHL6, SUDHL4, DB, WSUDLCL2, PFEIFFER), AML cell lines (OCIAML5), MDS cell lines (SKM1), ovarian cancer cell lines (OV7, TYKNU), esophageal cancer cell lines (KYSE150), rhabdoid tumor lines (RD, G402, G401, HS729, A204), liver cancer cell lines (HLF, HLE, PLCRPF5), and lung cancer cell lines (SW1573, NCIH2444) with the following modifications. Cells were seeded in 96-well plates, and the next day, BRG1 / BRM ATPase inhibitor Compound A was dissolved in DMSO and added to the cells in a concentration gradient of 0-10 μM. At the time of cell splitting on days 3 and 7, cells were split into new 96-well plates, and new compounds were added 4 hours after reseeding. Table 1 lists the test cell lines and growth media used. [Table 2]

[0177] Results: As shown in Figure 1, uveal melanoma and hematological cancer cell lines were more sensitive to BRG1 / BRM inhibition than the other cell lines tested. Inhibition of uveal melanoma and hematological cancer cell lines was maintained through day 7.

[0178] Example 8. Comparison of BRG1 / BRM inhibitors with clinical PKC and MEK inhibitors in uveal melanoma cell lines Procedure: Uveal melanoma cell lines, 92-1 or MP41, were seeded in 96-well plates in the presence of growth medium (see Table 1). A BAF ATPase inhibitor (Compound A), a PKC inhibitor (LXS196; MedChemExpress), or a MEK inhibitor (Selumetinib; Selleck Chemicals) were dissolved in DMSO and added to the cells at the time of seeding in a concentration gradient of 0–10 μM. Cells were incubated at 37°C for 3 days. After 3 days of treatment, cell proliferation was measured with a Cell-titer glow (Promega) and luminescence was read on an Envision plate reader (Perkin Elmer).

[0179] Results: As shown in Figures 2 and 3, Compound A exhibited comparable inhibition of uveal melanoma cell proliferation as clinical PKC and MEK inhibitors. Furthermore, Compound A was found to provide a faster onset of inhibition than clinical PKC and MEK inhibitors.

[0180] Example 9. Synthesis of Compound B The BRG1 / BRM inhibitor compound B has the following structure: [ka]

[0181] Compound B was synthesized as shown in Scheme 4 below. [ka]

[0182] To a mixture of (2S)-2-amino-4-methylsulfanyl-N-[4-[3-(4-pyridyl)phenyl]thiazol-2-yl]butanamide (2 g, 4.75 mmol, HCl salt) and 1-methylsulfonylpyrrole-3-carboxylic acid (898.81 mg, 4.75 mmol) in DMF (20 mL) was added EDCI (1.37 g, 7.13 mmol), HOBt (962.92 mg, 7.13 mmol), and DIEA (2.46 g, 19.00 mmol, 3.31 mL) and the mixture was stirred at 25° C. for 3 h. 2 2H2O (100 mL) and the precipitate was collected by filtration. The solid was triturated in MeOH (20 mL) and the precipitate was collected by filtration. The solid was dissolved in DMSO (10 mL) and the mixture was then poured into MeOH (50 mL) and the precipitate that formed was collected by filtration and lyophilized to give compound B (2.05 g, 3.66 mmol, 77.01% yield) as a white solid. LCMS(ESI)m / z[M+H] + =555.9. 1 H NMR(400MHz,DMSO)δ 12.49(s,1H),8.68-8.66(m,2H),8.46(d,J=7.2Hz,1H),8.31-8.30(m, 1H),8.02-8.00(m,1H),7.94-7.96(m,1H),7.83(s,1H),7.73-7.74(m,3 H),7.61-7.57(m,1H),7.31-7.29(m,1H),6.79-6.77(m,1H),4.74-4.6 9(m,1H),3.57(s,3H),2.67-2.53(m,2H),2.13-2.01(m,5H).ee%=100%.

[0183] Compound B had an IP of 3.6 nM for BRM and 5.7 nM for BRG1 in the ATPase assay described. 50 It was found to have the following structure:

[0184] Example 10. Effect of BRG1 / BRM ATPase inhibition on the proliferation of uveal melanoma, hematological cancer, prostate cancer, breast cancer, and Ewing's sarcoma cell lines Procedure: All cell lines listed above in Example 7 were also tested with Compound B as described above. In addition, the following cell lines were also tested as follows: Briefly, Ewing's sarcoma cell lines (CADOES1, RDES, SKES1), retinoblastoma cell line (WERIRB1), ALL cell line (REH), AML cell line (KASUMI1), prostate cancer cell lines (PC3, DU145, 22RV1), melanoma cell lines (SH4, SKMEL28, WM115, COLO829, SKMEL3, A375), breast cancer cell lines (MDAMB415, CAMA1, MCF7, BT474, HCC1419), The above method was performed for 1000 ng / ml IgG1-positive cells (IL-1, IL-2, DU4475, BT549), B-ALL cell lines (SUPB15), CML cell lines (K562, MEG01), Burkitt's lymphoma cell lines (RAMOS2G64C10, DAUDI), mantle cell lymphoma cell lines (JEKO1, REC1), bladder cancer cell lines (HT1197), and lung cancer cell lines (SBC5) with the following modifications: cells were seeded in 96-well plates, and the next day, BRG1 / BRM ATPase inhibitor, compound B, was dissolved in DMSO and added to the cells in a concentration gradient of 0-10 μM. At the time of cell splitting on days 3 and 7, cells were split into new 96-well plates, and new compounds were added 4 hours after reseeding. Table 2 lists the test cell lines and growth media used. [Table 3]

[0185] Results: As shown in Figure 4, uveal melanoma, hematological cancer, prostate cancer, breast cancer, and Ewing's sarcoma cell lines were more sensitive to BRG1 / BRM inhibition than the other cell lines tested. Inhibition of uveal melanoma, hematological cancer, prostate cancer, breast cancer, and Ewing's sarcoma cell lines was maintained through day 7.

[0186] Example 11. Effect of BRG1 / BRM ATPase inhibition on the proliferation of uveal melanoma and blood cancer cell lines Procedure: Pooled cell viability assays were performed using PRISM (simultaneous profiling of relative inhibition in mixtures) as previously described ("High-throughput identification of genotype-specific cancer vulnerabilities in mixtures of barcoded tumor cell lines," Yu et al, Nature Biotechnology 34, 419-423, 2016) with the following modifications. Cell lines were obtained from the Cancer Cell Line Encyclopedia (CCLE) collection and adapted to phenol red-free RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS) in order to apply the unique infection and pooling protocol to such a large list of cell lines. A lentiviral spin-infection protocol was performed to introduce the 24-nucleotide barcode into each cell line, with an estimated multiplicity of infection (MOI) of 1 for all cell lines, using blasticidin as a selection marker. Over 750 stably barcoded PRISM cancer cell lines were then pooled together into pools of 25 according to their doubling times. For screening runs, instead of seeding a pool of 25 cell lines in each well as previously described (Yu et al.), a pool of all adherent or all suspension cell lines was seeded together using T25 flasks (100,000 cells / flask) or 6-well plates (50,000 cells / well), respectively. Cells were treated with either DMSO or compounds in triplicate with 8-point, 3-fold dose-response starting at a top concentration of 10 μM. As a control for assay robustness, cells were treated in parallel with two previously validated compounds, the pan-Raf inhibitor AZ-628, and the proteasome inhibitor bortezomib, using top concentrations of 2.5 μM and 0.039 μM, respectively.

[0187] After 3 days of compound treatment, cells were lysed, genomic DNA was extracted, and barcodes were amplified by PCR and detected by next generation sequencing. Cell viability was determined by comparing the counts of cell line-specific barcodes in treated samples with those in DMSO and day 0 controls. Dose-response curves were fitted for each cell line and the corresponding area under the curve (AUC) was calculated and compared to the median AUC of all cell lines (Figure 5).

[0188] Results: Cell lines with AUC below the median were considered the most sensitive.

[0189] Example 12. Effect of BRG1 / BRM ATPase inhibitors on the proliferation of uveal melanoma cell lines Procedure: Uveal melanoma cell lines (92-1, MP41, MP38, MP46) and non-small cell lung cancer cells (NCIH1299) were seeded in 96-well plates containing growth medium (see Table 2). Compound B, a BRG1 / BRM ATPase inhibitor, was dissolved in DMSO and added to the cells at the time of seeding in a concentration gradient of 0-10 μM. Cells were incubated at 37°C for 3 days. After 3 days of treatment, cell proliferation was measured with a Cell-titer glow (Promega) and luminescence was read on an Envision plate reader (Perkin Elmer).

[0190] Results: As shown in FIG. 6, compound B produced potent growth inhibition in the cell lines.

[0191] Example 13. Comparison of BRG1 / BRM inhibitors with clinical PKC and MEK inhibitors in uveal melanoma cell lines Procedure: Uveal melanoma cell lines, 92-1 or MP41, were seeded in 96-well plates in the presence of growth medium (see Table 2). BAF ATPase inhibitor (Compound B), PKC inhibitor (LXS196; MedChemExpress), and MEK inhibitor (selumetinib; Selleck Chemicals) were dissolved in DMSO and added to cells at the time of seeding in a concentration gradient of 0-10 μM. Cells were incubated at 37°C for 3 days. After 3 days of treatment, cell proliferation was measured with a Cell-titer glow (Promega) and luminescence was read on an Envision plate reader (Perkin Elmer).

[0192] Results: As shown in Figures 7 and 8, Compound B showed a more potent effect on inhibiting the proliferation of uveal melanoma cells compared to clinical PKC inhibitors and clinical MEK inhibitors. Furthermore, Compound B was found to result in a faster onset of proliferation inhibition than clinical PKC inhibitors and clinical MEK inhibitors.

[0193] Example 14. BRG1 / BRM ATPase inhibitors are effective in inhibiting the proliferation of PKC inhibitor-resistant cells. Procedure: MP41 uveal melanoma cells were made resistant to a PKC inhibitor (LXS196, MedChemExpress) by long-term culture in growth medium containing increasing concentrations of the compound up to 1 μM (see Table 2). After 3 months, the sensitivity of parental MP41 cells and PKC inhibitor (PKCi)-resistant cells to the PKC inhibitor (LXS196) or the BRG1 / BRM ATPase inhibitor (Compound B) was tested in a 7-day growth inhibition assay as described above in Example 6.

[0194] Results: Although PKCi-resistant cells could tolerate growth at higher concentrations of LXS196 than the parental MP41 cell line (Figure 9), the BRG1 / BRM ATPase inhibitor (compound B) still resulted in strong growth inhibition of both the PKCi-resistant and parental cell lines (Figure 10). The PKCi-resistant cells were more sensitive to compound B than the parental MP41 cells (Figure 10).

[0195] Example 15. Synthesis of Compound C The BRG1 / BRM inhibitor compound C has the following structure: [ka]

[0196] Compound C was synthesized as shown in Scheme 5 below. [ka]

[0197] Compound C had an IP of 5.3 nM for BRM and 1.3 nM for BRG1 in the ATPase assay described above. 50 It was found to have the following structure:

[0198] Example 16. BRG1 / BRM ATPase inhibitors cause inhibition of uveal melanoma tumor growth in vivo. Procedure: Nude mice (Envigo) were cultured with 5 × 10 6 92-1 uveal melanoma cells were used to implant subcutaneously in the axillary region. Tumors grew to an average size of approximately 200 mm 3 The tumors were allowed to grow until the tumors were grown to 100% at which point the mice were grouped and dosing was initiated. Mice were dosed once daily by oral gavage with vehicle (20% 2-hydroxypropyl-β-cyclodextrin) or increasing doses of Compound C. Tumor volumes and body weights were measured over a 3-week period and doses were adjusted by body weight to obtain the appropriate dose in mg / kg. At this point the animals were euthanized and tumors were dissected and imaged.

[0199] Results: As shown in Figures 11 and 12, treatment with Compound C resulted in tumor growth inhibition with tumor regression observed at the highest (50 mg / kg) dose in a dose-dependent manner. As shown in Figure 13, both treatments were well tolerated and no weight loss was observed (Figure 13).

[0200] Example 17. Effect of BRG1 / BRM ATPase inhibition on the proliferation of uveal melanoma and blood cancer cell lines Procedure: Uveal melanoma cell lines (92-1, MEL202, MP41, MP38, MP46), prostate cancer cells (22RV1), acute leukemia cells (EOL1, THP1), and histiocytic lymphoma cells (U937) were seeded in 96-well plates containing growth medium (see Table 2). The BRG1 / BRM ATPase inhibitor, N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide, was dissolved in DMSO and added to the cells at the time of seeding in a concentration gradient of 0-2 μM (for uveal melanoma cell lines) or 0-1 μM (for other cell lines). The cells were incubated at 37 °C for 3 days. After 3 days of treatment, cell proliferation was measured by Cell-titer glow (Promega) and luminescence was read on an Envision plate reader (Perkin Elmer).

[0201] Results: As shown in FIG. 14, N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide produced potent growth inhibition in all cell lines. The measured absolute IC 50 Values ​​were below 350 nanomolar for all cell lines tested.

[0202] Table 3 shows the test cell lines and growth media used, as well as the absolute IC after 3 days of treatment with compounds. 50 List the values ​​(nM). [Table 4]

[0203] Example 18. BRG1 / BRM ATPase inhibitors cause inhibition of uveal melanoma tumor growth in vivo. Procedure: Nude mice (Envigo) were cultured with 5 × 10 6 92-1 uveal melanoma cells were used to implant subcutaneously in the axillary region. Tumors grew to an average size of approximately 200 mm 3 The tumors were allowed to grow until 12 h after tumor growth, at which point mice were grouped and dosing was initiated. Mice were dosed once daily by oral gavage with vehicle (20% 2-hydroxypropyl-β-cyclodextrin) or increasing doses of N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide. Tumor volumes and body weights were measured over a 3-week period and doses were adjusted by body weight to obtain the appropriate dose in mg / kg.

[0204] Results: As shown in FIG. 15, treatment with N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide resulted in tumor growth inhibition with tumor regression observed at the highest (1.5 mg / kg) dose in a dose-dependent manner. Both treatments were well tolerated based on the % body weight change observed, as shown in FIG. 16.

[0205] Example 19. Combination of FHD-286 and αPD-1 Ab provides synergistic benefit in the immunologically inactive B16F10 melanoma model. Procedure: B16F10 cells were implanted into mice and tumors were grown to 50 mm 3Mice were treated twice weekly with 1.5 mg / kg of N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide per day and 10 mg / kg of anti-PD-1 antibody.

[0206] Results: As shown in Figures 17-19, the combination of a BRM / BRG1 inhibitor and an anti-PD-1 antibody produced greater than additive effects on tumor inhibition and survival in this model.

[0207] Example 20. Combination of BRM / BRG1 inhibitors with PD-1 inhibitors provides synergistic benefit in A20 lymphoma tumor-bearing mice. Procedure: A20 cells were implanted into mice and tumors were grown to 50 mm 3 Mice were treated twice weekly with 1.5 mg / kg of N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide per day and 10 mg / kg of anti-PD-1 antibody.

[0208] Results: As shown in Figures 20-22, the combination of a BRM / BRG1 inhibitor and an anti-PD-1 antibody produced greater than additive effects on tumor inhibition and survival in this model.

[0209] Example 21. Combination of BRM / BRG1 inhibitors with PD-1 inhibitors provides synergistic benefit in CT26 colorectal tumor-bearing mice. Procedure: CT26 cells were implanted into mice and tumors were grown to 50 mm 3Mice were treated twice weekly with 1.5 mg / kg of N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide per day and 10 mg / kg of anti-PD-1 antibody.

[0210] Results: As shown in Figures 23-25, the combination of a BRM / BRG1 inhibitor with an anti-PD-1 antibody produced greater than additive effects on tumor inhibition and survival in this model.

[0211] Example 22. Combination of BRM / BRG1 inhibitors with PD-L1 inhibitors provides synergistic benefit in CT26 colorectal tumor-bearing mice. Procedure: CT26 cells were implanted into mice and tumors were grown to 50 mm 3 Mice were treated twice weekly with 1.5 mg / kg N-((S)-1-((4-(6-(cis-2,6-dimethylmorpholino)pyridin-2-yl)thiazol-2-yl)amino)-3-methoxy-1-oxopropan-2-yl)-1-(methylsulfonyl)-1H-pyrrole-3-carboxamide per day and 10 mg / kg anti-PD-L1 antibody.

[0212] Results: As shown in Figures 26 and 27, the combination of a BRM / BRG1 inhibitor with an anti-PD-L1 antibody produced greater than additive effects on tumor inhibition and survival in this model.

[0213] Other embodiments 1. A method of treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of an agent that reduces the level and / or activity of BRM and / or BRG1 and an effective amount of an immunotherapy. 2. A method of treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound having the following structure: [ka] or a pharma- ceutically acceptable salt thereof, and an effective amount of an immunotherapy. 3. The method of embodiment 1 or 2, wherein said immunotherapy is administered simultaneously with said agent or compound, or a pharma- ceutically acceptable salt thereof. 4. The method of embodiment 1 or 2, wherein said immunotherapy is administered prior to said agent or compound, or a pharma- ceutically acceptable salt thereof. 5. The method of embodiment 1 or 2, wherein said immunotherapy is administered after said agent or compound, or a pharma- ceutically acceptable salt thereof. 6. The method according to any one of embodiments 1 to 5, wherein said immunotherapy is a CTLA-4 inhibitor, a PD-1 inhibitor, a PD-L1 inhibitor, a CD-161 inhibitor, or an adoptive T cell transfer therapy. 7. The method of embodiment 6, wherein said immunotherapy is a CTLA-4 inhibitor. 8. The method of embodiment 6, wherein said immunotherapy is a PD-1 inhibitor. 9. The method of embodiment 6, wherein said immunotherapy is a PD-L1 inhibitor. 10. The method of embodiment 6, wherein said immunotherapy is a CD-161 inhibitor. 11. The method of embodiment 6, wherein said immunotherapy is adoptive T cell transfer therapy. 12. The method of any one of embodiments 1 to 11, wherein the cancer has not responded to a previously administered immunotherapy. 13. The method of any one of embodiments 1-12, wherein the cancer is resistant to immunotherapy. 14. The method of any one of embodiments 1 to 13, wherein the cancer does not contain a mutation that results in a loss of function of the BAF complex. 15. The method of any one of embodiments 1-14, wherein the effective amount of the agent or compound is an amount effective to increase the level of activated T cells in the subject. 16. The method of embodiment 15, wherein the effective amount of the agent or compound is an amount effective to increase the level of activated T cells in the tumor microenvironment. 17. The method of any one of embodiments 1-16, wherein the cancer is non-small cell lung cancer, colon cancer, bladder cancer, cancer of unknown primary, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, esophagogastric cancer, esophageal cancer, pancreatic cancer, hepatobiliary cancer, soft tissue sarcoma, ovarian cancer, head and neck cancer, renal cell carcinoma, bone cancer, non-Hodgkin's lymphoma, small cell lung cancer, prostate cancer, embryonal tumors, germ cell tumors, cervical cancer, thyroid cancer, salivary gland cancer, gastrointestinal neuroendocrine tumors, uterine sarcoma, gastrointestinal stromal tumors, CNS cancer, thymic tumors, adrenocortical carcinoma, appendix cancer, small intestine cancer, penile cancer, bone cancer, or blood cancer. 18. The method of embodiment 17, wherein the cancer is esophageal cancer. 19. The method of embodiment 18, wherein the cancer is non-small cell lung cancer, colon cancer, bladder cancer, cancer of unknown primary, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, penile cancer, bone cancer, renal cell carcinoma, prostate cancer, or blood cancer. 20. The method of embodiment 19, wherein the cancer is non-small cell lung cancer. 21. The method of embodiment 19, wherein the cancer is melanoma, prostate cancer, breast cancer, bone cancer, renal cell carcinoma, or blood cancer. 22. The method of embodiment 21, wherein the cancer is melanoma. 23. The method of embodiment 22, wherein the melanoma is uveal melanoma, mucosal melanoma, or cutaneous melanoma. 24. The method of embodiment 23, wherein the melanoma is uveal melanoma. 25. The method of embodiment 21, wherein the cancer is prostate cancer. 26. The method of embodiment 21, wherein the cancer is a blood cancer. 27. The method of embodiment 26, wherein the hematological cancer is multiple myeloma, large cell lymphoma, acute T-cell leukemia, acute myeloid leukemia, myelodysplastic syndrome, immunoglobulin A lambda myeloma, diffuse mixed histiocytic and lymphocytic lymphoma, B-cell lymphoma, acute lymphoblastic leukemia, diffuse large cell lymphoma, or non-Hodgkin's lymphoma. 28. The method of embodiment 21, wherein the cancer is breast cancer. 29. The method of embodiment 28, wherein the breast cancer is ER-positive breast cancer, ER-negative breast cancer, triple-positive breast cancer, or triple-negative breast cancer. 30. The method of embodiment 21, wherein the cancer is bone cancer. 31. The method of embodiment 30, wherein the bone cancer is Ewing's sarcoma. 32. The method of embodiment 21, wherein the cancer is renal cell carcinoma. 33. The method of embodiment 32, wherein the renal cell carcinoma is microphthalmia transcription factor family translocation renal cell carcinoma. 34. The method of any one of embodiments 1 to 33, wherein the cancer expresses BRG1 and / or BRM proteins. 35. The method of any one of embodiments 1-34, wherein the subject or cancer has and / or has been identified as having a loss-of-function mutation in BRG1. 36. The method of embodiment 35, wherein the BRG1 loss-of-function mutation is in the ATPase catalytic domain of the protein. 37. The method of embodiment 35, wherein the loss-of-function mutation of BRG1 is a deletion at the C-terminus of BRG1. 38. The method of any one of embodiments 1 to 37, wherein the cancer does not have, or has been determined to not have, an epidermal growth factor receptor mutation and / or an anaplastic lymphoma kinase driver mutation. 39. The method of any one of embodiments 1 to 38, wherein the cancer has or is determined to have a KRAS mutation, a GNAQ mutation, a GNA11 mutation, a PLCB4 mutation, a CYSLTR2 mutation, a BAP1 mutation, a SF3B1 mutation, an EIF1AX mutation, a TFE3 rearrangement, a TFEB rearrangement, a MITF rearrangement, an EZH2 mutation, a SUZ12 mutation, and / or an EED mutation. 40. The method of any one of embodiments 1-39, wherein the cancer is metastatic. 41. The method of any one of embodiments 1 to 40, wherein the cancer is resistant to an anti-cancer therapy or has not responded to previous treatment with an anti-cancer therapy. 42. The method of embodiment 41, wherein the anticancer therapy is a chemotherapeutic or cytotoxic agent, immunotherapy, surgery, radiation therapy, hyperthermia, or photocoagulation, or a combination thereof. 43. The method of embodiment 42, wherein the anti-cancer therapy is a chemotherapeutic or cytotoxic agent. 44. The method of embodiment 43, wherein the chemotherapeutic or cytotoxic agent is a mitogen-activated protein kinase (MEK) inhibitor and / or a protein kinase C (PKC) inhibitor. 45. The method of any one of embodiments 1 to 44, wherein the cancer is resistant to a PKC inhibitor or has not responded to previous treatment with a PKC inhibitor. 46. ​​The method of any one of embodiments 1 to 45, wherein the method further comprises administering to the subject or contacting the cells with an anti-cancer therapy. 47. The method of embodiment 46, wherein the anticancer therapy is a chemotherapeutic or cytotoxic agent, immunotherapy, surgery, radiation therapy, hyperthermia, or photocoagulation, or a combination thereof. 48. The method of embodiment 46 or 47, wherein the anticancer therapy is surgery, a MEK inhibitor, and / or a PKC inhibitor, or a combination thereof. 49. The method of embodiment 48, wherein the MEK inhibitor is selumetinib, binimetinib, or tametinib. 50. The method of embodiment 48, wherein the PKC inhibitor is sotrastaurin or IDE 196. 51. An agent that reduces the level and / or activity of BRM and / or BRG1 for use in combination with immunotherapy for treating cancer in a subject in need thereof. 52. A compound having the following structure for use in combination with immunotherapy to treat cancer in a subject in need thereof: [ka] or a pharma- ceutically acceptable salt thereof. 53. Use of an agent that reduces the level and / or activity of BRM and / or BRG1 in the manufacture of a medicament for use in combination with immunotherapy for treating cancer in a subject in need thereof. 54. A compound having the following structure in the manufacture of a medicament for use in combination with immunotherapy to treat cancer in a subject in need thereof: [ka] or a pharma- ceutically acceptable salt thereof.

[0214] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference in their entirety to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference in its entirety. In the event that a term in this application is found to be defined differently in a document incorporated herein by reference, the definition provided herein shall serve as the definition of that term.

[0215] While the invention has been described in connection with specific embodiments thereof, it will be understood that the invention is capable of further modifications, and this application is intended to cover any variations, uses, or adaptations of the invention which generally follow the principles of the invention and include departures from the present disclosure which come within known or customary practice within the art to which the invention pertains, and may be applied to the essential features set forth herein above, and which fall within the scope of the claims.

Claims

1. A pharmaceutical composition comprising an agent that reduces the level and / or activity of BRM and / or BRG1, in an effective amount, for use in combination with an effective amount of immunotherapy in the treatment of cancer in a subject in need of treating cancer.

2. A pharmaceutical composition comprising an effective amount of a compound having the following structure, for use in combination with an effective amount of immunotherapy in the treatment of cancer in a subject in need of treating cancer. 【Chemical 1】 Or a pharmaceutically acceptable salt thereof.

3. The immunotherapy is (a) administered simultaneously with the agent or the compound, or a pharmaceutically acceptable salt thereof; (b) administered before the agent or the compound, or a pharmaceutically acceptable salt thereof; or (c) administered after the agent or the compound, or a pharmaceutically acceptable salt thereof. The pharmaceutical composition according to claim 1 or 2, formulated for

4. The immunotherapy is a PD-1 inhibitor, a CTLA-4 inhibitor, a PD-L1 inhibitor, a CD-161 inhibitor, or adoptive T cell transfer therapy. The pharmaceutical composition according to any one of claims 1 to 3.

5. The cancer is (a) not responsive to a previously administered immunotherapy; (b) resistant to immunotherapy; and / or (c) does not contain a mutation that results in loss of function of the BAF complex. The pharmaceutical composition according to any one of claims 1 to 4.

6. The effective amount of the agent or the compound is (a) effective to increase the level of activated T cells in the subject; and / or (b) effective to increase the level of activated T cells in the tumor microenvironment. The pharmaceutical composition according to any one of claims 1 to 5.

7. The cancer is non-small cell lung cancer, colorectal cancer, bladder cancer, cancer of unknown primary origin, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, esophageal gastric cancer, esophageal cancer, pancreatic cancer, hepatobiliary tract cancer, soft tissue sarcoma, ovarian cancer, head and neck cancer, renal cell cancer, bone cancer, non-Hodgkin lymphoma, small cell lung cancer, prostate cancer, fetal tumor, germ cell tumor, cervical cancer, thyroid cancer, salivary gland cancer, gastrointestinal neuroendocrine tumor, uterine sarcoma, gastrointestinal stromal tumor, CNS cancer, thymic tumor, adrenocortical carcinoma, appendiceal cancer, small intestine cancer, penile cancer, bone cancer, or blood cancer. The pharmaceutical composition according to any one of claims 1 to 6.

8. (a) The melanoma is uveal melanoma, mucosal melanoma, or cutaneous melanoma. (b) the blood cancer is multiple myeloma, large cell lymphoma, acute T cell leukemia, acute myeloid leukemia, myelodysplastic syndrome, immunoglobulin A lambda myeloma, diffuse mixed histiocytic lymphoma and lymphocytic lymphoma, B cell lymphoma, acute lymphoblastic leukemia, diffuse large B cell lymphoma, or non-Hodgkin lymphoma, (c) the breast cancer is ER-positive breast cancer, ER-negative breast cancer, triple-positive breast cancer, or triple-negative breast cancer, (d) the bone cancer is Ewing sarcoma, or (e) the renal cell cancer is small eye transcription factor family translocation renal cell cancer, The pharmaceutical composition according to claim 7.

9. (a) the cancer expresses BRG1 and / or BRM protein, (b) the subject or the cancer has a loss-of-function mutation of BRG1, (c) the cancer does not have or is determined not to have an epidermal growth factor receptor mutation and / or an anaplastic lymphoma kinase driver mutation, (d) the cancer has or is determined to have a KRAS mutation, GNAQ mutation, GNA11 mutation, PLCB4 mutation, CYSLTR2 mutation, BAP1 mutation, SF3B1 mutation, EIF1AX mutation, TFE3 translocation, TFEB translocation, MITF translocation, EZH2 mutation, SUZ12 mutation, and / or EED mutation, (e) the cancer is metastatic, (f) the cancer is resistant to anticancer therapy or did not respond to previous treatment with anticancer therapy, and / or (g) the cancer is resistant to a PKC inhibitor or did not respond to previous treatment with a PKC inhibitor, The pharmaceutical composition according to any one of claims 1 to 8.

10. The loss-of-function mutation of BRG1 is (a) in the ATPase catalytic domain of the protein, or (b) a deletion at the C-terminus of BRG1, The pharmaceutical composition according to claim 9.

11. The anticancer therapy is a chemotherapeutic agent or cytotoxic agent, immunotherapy, surgery, radiotherapy, hyperthermia, or photocoagulation, or a combination thereof, the pharmaceutical composition according to claim 9.

12. The chemotherapeutic agent or cytotoxic agent is a mitogen-activated protein kinase (MEK) inhibitor and / or a protein kinase C (PKC) inhibitor, the pharmaceutical composition according to claim 11.

13. The pharmaceutical composition according to any one of claims 1 to 12, wherein the compound or the agent is formulated for use in combination with an anti-cancer therapy.

14. The anti-cancer therapy is (a) a chemotherapeutic agent or cytotoxic agent, immunotherapy, surgery, radiotherapy, hyperthermia, or photocoagulation, or a combination thereof, or (b) surgery, a MEK inhibitor, and / or a PKC inhibitor, or a combination thereof, The pharmaceutical composition according to claim 13.

15. (a) The MEK inhibitor is selumetinib, binimetinib, or trametinib, or (b) The PKC inhibitor is sotrastaurin or IDE196, The pharmaceutical composition according to claim 14.

16. Use of an agent that reduces the level and / or activity of BRM and / or BRG1 in the manufacture of a medicament for use in combination with an immunotherapy for treating cancer in a subject in need of treating cancer.

17. Use of a compound having the following structure: [Chemical 2] or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for use in combination with an immunotherapy for treating cancer in a subject in need of treating cancer.

18. The use according to claim 16 or 17, wherein the immunotherapy is a PD-1 inhibitor, a CTLA-4 inhibitor, a PD-L1 inhibitor, a CD-161 inhibitor, or adoptive T cell transfer therapy.

19. (a) The cancer did not respond to a previously administered immunotherapy, (b) The cancer is resistant to immunotherapy, (c) The cancer does not contain a mutation that results in loss of function of the BAF complex, (d) The cancer expresses BRG1 and / or BRM protein, (e) The subject or the cancer has a loss-of-function mutation in BRG1, (f) The cancer is determined not to have or not to have an epidermal growth factor receptor mutation and / or an anaplastic lymphoma kinase driver mutation, (g) The cancer is determined to have or have a KRAS mutation, a GNAQ mutation, a GNA11 mutation, a PLCB4 mutation, a CYSLTR2 mutation, a BAP1 mutation, an SF3B1 mutation, an EIF1AX mutation, a TFE3 translocation, a TFEB translocation, a MITF translocation, an EZH2 mutation, a SUZ12 mutation, and / or an EED mutation, (h) The cancer is metastatic, (i) the cancer is resistant to anti-cancer therapy or did not respond to previous treatment with anti-cancer therapy, and / or (j) the cancer is resistant to a PKC inhibitor or did not respond to previous treatment with a PKC inhibitor Use according to any one of claims 16 to 18.

20. Use according to any one of claims 16 to 19, wherein the cancer is non-small cell lung cancer, colorectal cancer, bladder cancer, cancer of unknown primary origin, glioma, breast cancer, melanoma, non-melanoma skin cancer, endometrial cancer, esophagogastric cancer, esophageal cancer, pancreatic cancer, hepatobiliary tract cancer, soft tissue sarcoma, ovarian cancer, head and neck cancer, renal cell cancer, bone cancer, non-Hodgkin lymphoma, small cell lung cancer, prostate cancer, fetal tumor, germ cell tumor, cervical cancer, thyroid cancer, salivary gland cancer, gastrointestinal neuroendocrine tumor, uterine sarcoma, gastrointestinal stromal tumor, CNS cancer, thymic tumor, adrenocortical carcinoma, appendiceal cancer, small intestine cancer, penile cancer, bone cancer, or blood cancer.

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