Compounds and their uses

Abstract

The present disclosure relates to compounds useful for treating BAF complex-associated disorders.

Description

[Technical field]

[0001] The present invention relates to compounds and uses thereof. [Background technology]

[0002] The disorder may be affected by the BAF complex. BRD9 is a component of the BAF complex. The present invention relates to compositions and methods useful for treating BAF complex-associated disorders, such as cancer and infectious diseases. Summary of the Invention [Means for solving the problem]

[0003] Bromodomain-containing protein 9 (BRD9) is a protein encoded by the BRD9 gene on chromosome 5. BRD9 is a component of the BAF (BRG1 or BRM-associated factor) complex, a SWI / SNF ATPase chromatin remodeling complex, and belongs to family IV of bromodomain-containing proteins. BRD9 is present in several SWI / SNF ATPase chromatin remodeling complexes and is upregulated in several cancer cell lines. Thus, agents that reduce the level and / or activity of BRD9 may provide new methods for treating diseases and disorders, such as cancer and infectious diseases. The inventors have discovered that depleting BRD9 in cells results in depletion of SS18-SSX fusion proteins in those cells. SS18-SSX fusion proteins have been detected in more than 95% of synovial sarcoma tumors and are often the only chromosomal abnormality in synovial sarcoma. In addition, evidence suggests that the BAF complex is involved in the antiviral activity of cells. Thus, agents (eg, compounds) that degrade BRD9 are useful in treating disorders (eg, cancer or infectious diseases) associated with BAF, BRD9, and / or SS18-SSX.

[0004] The present disclosure relates to compounds and methods useful for treating BAF-associated disorders (eg, cancer or infectious diseases). In one aspect, the invention relates to a compound having the structure of any one of compounds D1, S-D1, R-D1, and D2 in Table 1, or a pharma- ceutically acceptable salt thereof.

[0005] In some embodiments, the compound has the structure of compound D1, or is a pharma- ceutically acceptable salt thereof. In some embodiments, the compound has the structure of compound S-D1, or is a pharma- ceutically acceptable salt thereof. In some embodiments, the compound has the structure of compound R-D1, or is a pharma- ceutically acceptable salt thereof. In other embodiments, the compound has the structure of compound D2, or is a pharma- ceutically acceptable salt thereof.

[0006] In one embodiment, the invention relates to a compound having the structure of Compound D1 in Table 1, or a pharma- ceutically acceptable salt thereof. In one embodiment, the invention relates to a compound having the structure of compound S-D1 in Table 1, or a pharma- ceutically acceptable salt thereof.

[0007] In one embodiment, the invention relates to a compound having the structure of compound R-D1 in Table 1, or a pharma- ceutically acceptable salt thereof. In another aspect, the invention relates to a compound having the structure of Compound D2 in Table 1, or a pharma- ceutically acceptable salt thereof.

[0008] [Table 1]

[0009] In another aspect, the present disclosure relates to a pharmaceutical composition comprising any of the aforementioned compounds, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable excipient. In one aspect, the disclosure relates to a method for inhibiting the level and / or activity of BRD9 in a cell, the method comprising contacting the cell with an effective amount of any of the aforementioned compounds, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0010] In another aspect, the disclosure relates to a method for reducing the level and / or activity of BRD9 in a cell, the method comprising contacting the cell with an effective amount of any of the aforementioned compounds, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0011] In some embodiments, the cell is a cancer cell. In some embodiments, the cancer is a malignant tumor, a rhabdoid tumor, a CD8+ T-cell lymphoma, an endometrial cancer, an ovarian cancer, a bladder cancer, a gastric cancer, a pancreatic cancer, an esophageal cancer, a prostate cancer, a renal cell carcinoma, a melanoma, a colorectal cancer, a sarcoma (e.g., a soft tissue sarcoma, a synovial sarcoma, an Ewing's sarcoma, an osteosarcoma, an rhabdomyosarcoma, an adult fibrosarcoma, an alveolar soft part sarcoma, angiosarcoma, a clear cell sarcoma, a desmoplastic small round cell tumor, an epithelioid sarcoma, an fibromyxoid sarcoma, an gastrointestinal stromal tumor, a Kaposi's sarcoma, a liposarcoma, a leiomyosarcoma, an malignant mesenchymal tumor, an malignant peripheral nerve sheath tumor, an myxofibrosarcoma, an angio ... In some embodiments, the cancer is malignant tumor, rhabdoid tumor, CD8+T cell lymphoma, endometrial cancer, ovarian cancer, bladder cancer, gastric cancer, pancreatic cancer, esophageal cancer, prostate cancer, renal cell carcinoma, melanoma, or colorectal cancer.In some embodiments, the cancer is sarcoma (e.g., synovial sarcoma or Ewing's sarcoma), non-small cell lung cancer (e.g., squamous cell or adenocarcinoma), gastric cancer, or breast cancer.In some embodiments, the cancer is sarcoma (e.g., synovial sarcoma or Ewing's sarcoma).In some embodiments, the sarcoma is synovial sarcoma.

[0012] In some aspects, the present disclosure relates to a method of treating a BAF complex-associated disorder in a subject in need thereof, comprising administering to the subject an effective amount of any of the aforementioned compounds, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the BAF complex-associated disorder is cancer. In some embodiments, the BAF complex-associated disorder is an infectious disease.

[0013] In another aspect, the present disclosure relates to a method of treating a SS18-SSX fusion protein-associated disorder in a subject in need thereof, comprising administering to the subject an effective amount of any of the aforementioned compounds, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the SS18-SSX fusion protein-associated disorder is cancer. In some embodiments, the SS18-SSX fusion protein-associated disorder is an infectious disease. In some embodiments of any of the aforementioned methods, the SS18-SSX fusion protein is a SS18-SSX1 fusion protein, a SS18-SSX2 fusion protein, or a SS18-SSX4 fusion protein.

[0014] In yet another aspect, the present disclosure relates to a method of treating a BRD9-related disorder in a subject in need thereof, comprising administering to the subject an effective amount of any of the aforementioned compounds, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the BRD9-related disorder is cancer. In some embodiments, the BRD9-related disorder is an infectious disease.

[0015] In some embodiments, the cancer is selected from the group consisting of squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, hepatocellular carcinoma, and renal cell carcinoma, cancer of the bladder, intestine, breast, cervix, colon, esophagus, head, kidney, liver, lung, neck, ovary, pancreas, prostate, and stomach; leukemia; benign and malignant lymphomas, particularly Burkitt's lymphoma and non-Hodgkin's lymphoma; benign and malignant melanoma; myeloproliferative disorders; sarcomas, such as Ewing's sarcoma, angiosarcoma, Kaposi's sarcoma, liposarcoma, myosarcoma, peripheral neuroepithelioma, synovial sarcoma, glioma, astrocytoma, oligodendroglioma, ependymoma, gliobastromas, neuroblastoma, ganglioneuroma, ganglioglioma, medulloblastoma, pinealocyte tumor, meningioma, meningeal sarcoma, neurofibroma, and schwannoma; intestinal cancer, breast cancer, prostate cancer, cervical cancer, uterine cancer, lung cancer, ovarian cancer, testicular cancer, thyroid cancer, astrocytoma, esophageal cancer, pancreatic cancer, gastric cancer, liver cancer, colon cancer, melanoma; carcinosarcoma, Hodgkin's disease, Wilms' tumor, and teratocarcinoma. Additional cancers that may be treated using the disclosed compounds according to the present invention include, for example, acute granulocytic leukemia, acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), adenocarcinoma, adenosarcoma, adrenal gland cancer, adrenal cortical carcinoma, anal cancer, anaplastic astrocytoma, angiosarcoma, appendix cancer, astrocytoma, basal cell carcinoma, B cell lymphoma, bile duct cancer, bladder cancer, bone cancer, bone marrow cancer, intestinal cancer, brain cancer, brain stem glioma, breast cancer, triple (estrogen, progesterone and HER-2) negative breast cancer, double negative breast cancer (two of estrogen, progesterone and HER-2 are negative), single negative (one of estrogen, progesterone and HER-2 is negative), estrogen receptor positive breast cancer, and / or other cancers that may be treated using the disclosed compounds according to the present invention. HER2 negative breast cancer, estrogen receptor negative breast cancer, estrogen receptor positive breast cancer, metastatic breast cancer, luminal A breast cancer, luminal B breast cancer, Her2 negative breast cancer, HER2 positive or negative breast cancer, progesterone receptor negative breast cancer, progesterone receptor positive breast cancer, recurrent breast cancer, carcinoid tumor, cervical cancer, bile duct cancer, chondrosarcoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), colon cancer, colorectal cancer, craniopharyngioma, cutaneous lymphoma, cutaneous melanoma, diffuse astrocytoma, ductal carcinoma in situ (DCIS), endometrial cancer, ependymoma, epithelioid sarcoma, esophageal cancer, Ewing's sarcoma, extrahepatic bile duct cancer, eye cancer, fallopian tube cancer, fibrosarcoma, gallbladder cancer, gastric cancer,Gastrointestinal cancer, gastrointestinal carcinoid cancer, gastrointestinal stromal tumor (GIST), germ cell tumor glioblastoma multiforme (GBM), glioma, hairy cell leukemia, head and neck cancer, hemangioendothelioma, Hodgkin's lymphoma, hypopharyngeal cancer, invasive ductal carcinoma (IDC), invasive lobular carcinoma (ILC), inflammatory breast cancer (IBC), intestinal cancer, intrahepatic cholangiocarcinoma, invasive / invasive breast cancer, islet cell carcinoma, jaw cancer, Kaposi's sarcoma, kidney cancer, laryngeal cancer, leiomyosarcoma, leptomeningeal metastasis, leukemia, lip cancer, liposarcoma, liver cancer, lobular carcinoma in situ, low-grade astrocytoma, lung cancer, lymph node cancer, lymphoma, male breast cancer, medullary carcinoma, medulloblastoma, melanoma, meningioma, Merkel cell carcinoma tumor, mesenchymal chondrosarcoma, mesenchymal, mesothelioma, metastatic breast cancer, metastatic melanoma, metastatic squamous cell neck cancer, mixed glioma, monodermal teratoma, oral cancer, mucinous carcinoma, mucinous melanoma, multiple myeloma, mycosis fungoides, myelodysplastic syndrome, nasal cancer, nasopharyngeal cancer, cervical cancer, neuroblastoma, neuroendocrine tumor (NET), non-Hodgkin's lymphoma, non-small cell lung cancer (NSCLC), oat cell carcinoma, ocular cancer, ocular melanoma, oligodendroglioma, oral cancer, oral cancer, oropharyngeal cancer, osteogenic sarcoma, osteosarcoma, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, ovarian primary peritoneal cancer, ovarian sex cord stromal tumor, Paget's disease, pancreatic cancer, papillary cancer, paranasal sinus cancer, parathyroid cancer, pelvic cancer, penile cancer, peripheral nerve cancer, peritoneal cancer, pharyngeal cancer, pheochromocytoma, pilocytic astrocytoma, pineal region tumor, pineoblastoma, pituitary cancer, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, renal cell carcinoma, renal pelvis cancer, rhabdomyosarcoma, salivary gland cancer, soft tissue sarcoma, osteosarcoma, sarcoma, sinus cancer, skin cancer, small cell lung cancer (SCLC), small intestine cancer, spinal column cancer, spinal cord cancer, squamous cell carcinoma, gastric cancer, synovial sarcoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma / thymic carcinoma, thyroid cancer, tongue cancer, tonsil cancer, transitional cell carcinoma, tubular carcinoma, tubular carcinoma, undiagnosed cancer, ureter cancer , urethral cancer, uterine adenocarcinoma, uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, T-cell acute lymphoblastic leukemia (T-ALL), T-cell lymphoblastic lymphoma (T-LL), peripheral T-cell lymphoma, adult T-cell leukemia, precursor B-cell ALL, precursor B-cell lymphoma, large B-cell lymphoma, Burkitt lymphoma, B-cell ALL, Philadelphia chromosome positive ALL, Philadelphia chromosome positive CML, juvenile myelomonocytic leukemia (JMML), acute promyelocytic leukemia (subtype of AML), large granular lymphocyte leukemia, adult T-cell chronic leukemia, diffuse large B-cell lymphoma,Follicular lymphoma; mucosa-associated lymphoid tissue lymphoma (MALT), small cell lymphoma, mediastinal large B-cell lymphoma, nodal marginal zone B-cell lymphoma (NMZL); splenic marginal zone lymphoma (SMZL); intravascular large B-cell lymphoma; primary effusion lymphoma; or lymphomatoid granulomatosis; B-cell prolymphocytic leukemia; splenic lymphoma / leukemia, unclassifiable, splenic diffuse red pulp small B-cell lymphoma; lymphoplasmacytic lymphoma; heavy chain disease, e.g., alpha heavy chain disease, gamma heavy chain disease, mu heavy chain disease, plasma cell myeloma, solitary plasmacytoma of bone; extraosseous plasmacytoma; primary cutaneous follicular center lymphoma, T cell These include: follicular / histiocytic-rich large B-cell lymphoma, chronic inflammation-associated DLBCL; age-related Epstein-Barr virus (EBV)-positive DLBCL; primary mediastinal (thymic) large B-cell lymphoma, primary cutaneous DLBCL, leg type, ALK-positive large B-cell lymphoma, plasmablastic lymphoma; large B-cell lymphoma arising in HHV8-associated multicentric Castleman disease; B-cell lymphoma with features intermediate between diffuse large B-cell lymphoma, unclassified, or B-cell lymphoma with features intermediate between diffuse large B-cell lymphoma and classical Hodgkin lymphoma, unclassified.

[0016] In some embodiments, the cancer is a malignant tumor, a rhabdoid tumor, a CD8+ T-cell lymphoma, an endometrial cancer, an ovarian cancer, a bladder cancer, a gastric cancer, a pancreatic cancer, an esophageal cancer, a prostate cancer, a renal cell carcinoma, a melanoma, a colorectal cancer, a sarcoma (e.g., a soft tissue sarcoma, a synovial sarcoma, an Ewing's sarcoma, an osteosarcoma, an rhabdomyosarcoma, an adult fibrosarcoma, an alveolar soft part sarcoma, angiosarcoma, a clear cell sarcoma, a desmoplastic small round cell tumor, an epithelioid sarcoma, an fibromyxoid sarcoma, an gastrointestinal stromal tumor, a Kaposi's sarcoma, a liposarcoma, a leiomyosarcoma, an malignant mesenchymal tumor, an malignant peripheral nerve sheath tumor, an myxofibrosarcoma, an angio ... In some embodiments, the cancer is malignant tumor, rhabdoid tumor, CD8+T cell lymphoma, endometrial cancer, ovarian cancer, bladder cancer, gastric cancer, pancreatic cancer, esophageal cancer, prostate cancer, renal cell carcinoma, melanoma, or colorectal cancer.In some embodiments, the cancer is sarcoma (e.g., synovial sarcoma or Ewing's sarcoma), non-small cell lung cancer (e.g., squamous cell or adenocarcinoma), gastric cancer, or breast cancer.In some embodiments, the cancer is sarcoma (e.g., synovial sarcoma or Ewing's sarcoma).In some embodiments, the sarcoma is synovial sarcoma.

[0017] In some embodiments, the infection is a viral infection (e.g., Retroviridae family, such as lentiviruses (e.g., human immunodeficiency virus (HIV) and delta retroviruses (e.g., human T-cell leukemia virus I (HTLV-I), human T-cell leukemia virus II (HTLV-II)); Hepadnaviridae family (e.g., hepatitis B virus (HBV)); Flaviviridae family (e.g., hepatitis C virus (HCV)); Adenoviridae family (e.g., human adenovirus); Herpesviridae family (e.g., human cytomegalovirus (HCMV), Epstein-Barr virus, herpes simplex virus 1 (HSV-1), herpes simplex virus 2 (HSV-2), human herpes virus 6 (HHV-6), herpes vitus virus K, herpes viridans ... *, CMV, Varicella-Zoster Virus); Papillomaviridae family (e.g., Human Papillomavirus (HPV, HPV E1)); Parvoviridae family (e.g., Parvovirus B19); Polyomaviridae family (e.g., JC virus and BK virus); Paramyxoviridae family (e.g., Measles virus); or Togaviridae family (e.g., Rubella virus). In some embodiments, the disorder is coffin siris, neurofibromatosis (e.g., NF-1, NF-2, or Schwannomatosis), or multiple meningiomas. In some aspects, the disclosure relates to a method of treating cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of any of the aforementioned compounds, or a pharmacologic acceptable salt thereof, or any of the aforementioned pharmaceutical compositions.

[0018] In some embodiments, the cancer is selected from the group consisting of squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, hepatocellular carcinoma, and renal cell carcinoma, cancer of the bladder, intestine, breast, cervix, colon, esophagus, head, kidney, liver, lung, neck, ovary, pancreas, prostate, and stomach; leukemia; benign and malignant lymphomas, particularly Burkitt's lymphoma and non-Hodgkin's lymphoma; benign and malignant melanoma; myeloproliferative disorders; sarcomas, such as Ewing's sarcoma, angiosarcoma, Kaposi's sarcoma, liposarcoma, myosarcoma, peripheral neuroepithelioma, synovial sarcoma, glioma, astrocytoma, oligodendroglioma, ependymoma, gliobastromas, neuroblastoma, ganglioneuroma, ganglioglioma, medulloblastoma, pinealocyte tumor, meningioma, meningeal sarcoma, neurofibroma, and schwannoma; intestinal cancer, breast cancer, prostate cancer, cervical cancer, uterine cancer, lung cancer, ovarian cancer, testicular cancer, thyroid cancer, astrocytoma, esophageal cancer, pancreatic cancer, gastric cancer, liver cancer, colon cancer, melanoma; carcinosarcoma, Hodgkin's disease, Wilms' tumor, and teratocarcinoma. Additional cancers that may be treated using the disclosed compounds according to the present invention include, for example, acute granulocytic leukemia, acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), adenocarcinoma, adenosarcoma, adrenal gland cancer, adrenal cortical carcinoma, anal cancer, anaplastic astrocytoma, angiosarcoma, appendix cancer, astrocytoma, basal cell carcinoma, B cell lymphoma, bile duct cancer, bladder cancer, bone cancer, bone marrow cancer, intestinal cancer, brain cancer, brain stem glioma, breast cancer, triple (estrogen, progesterone and HER-2) negative breast cancer, double negative breast cancer (two of estrogen, progesterone and HER-2 are negative), single negative (one of estrogen, progesterone and HER-2 is negative), estrogen receptor positive breast cancer, and / or other cancers that may be treated using the disclosed compounds according to the present invention. HER2 negative breast cancer, estrogen receptor negative breast cancer, estrogen receptor positive breast cancer, metastatic breast cancer, luminal A breast cancer, luminal B breast cancer, Her2 negative breast cancer, HER2 positive or negative breast cancer, progesterone receptor negative breast cancer, progesterone receptor positive breast cancer, recurrent breast cancer, carcinoid tumor, cervical cancer, bile duct cancer, chondrosarcoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), colon cancer, colorectal cancer, craniopharyngioma, cutaneous lymphoma, cutaneous melanoma, diffuse astrocytoma, ductal carcinoma in situ (DCIS), endometrial cancer, ependymoma, epithelioid sarcoma, esophageal cancer, Ewing's sarcoma, extrahepatic bile duct cancer, eye cancer, fallopian tube cancer, fibrosarcoma, gallbladder cancer, gastric cancer,Gastrointestinal cancer, gastrointestinal carcinoid cancer, gastrointestinal stromal tumor (GIST), germ cell tumor glioblastoma multiforme (GBM), glioma, hairy cell leukemia, head and neck cancer, hemangioendothelioma, Hodgkin's lymphoma, hypopharyngeal cancer, invasive ductal carcinoma (IDC), invasive lobular carcinoma (ILC), inflammatory breast cancer (IBC), intestinal cancer, intrahepatic cholangiocarcinoma, invasive / invasive breast cancer, islet cell carcinoma, jaw cancer, Kaposi's sarcoma, kidney cancer, laryngeal cancer, leiomyosarcoma, leptomeningeal metastasis, leukemia, lip cancer, liposarcoma, liver cancer, lobular carcinoma in situ, low-grade astrocytoma, lung cancer, lymph node cancer, lymphoma, male breast cancer, medullary carcinoma, medulloblastoma, melanoma, meningioma, Merkel cell carcinoma tumor, mesenchymal chondrosarcoma, mesenchymal, mesothelioma, metastatic breast cancer, metastatic melanoma, metastatic squamous cell neck cancer, mixed glioma, monodermal teratoma, oral cancer, mucinous carcinoma, mucinous melanoma, multiple myeloma, mycosis fungoides, myelodysplastic syndrome, nasal cancer, nasopharyngeal cancer, cervical cancer, neuroblastoma, neuroendocrine tumor (NET), non-Hodgkin's lymphoma, non-small cell lung cancer (NSCLC), oat cell carcinoma, ocular cancer, ocular melanoma, oligodendroglioma, oral cancer, oral cancer, oropharyngeal cancer, osteogenic sarcoma, osteosarcoma, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, ovarian primary peritoneal cancer, ovarian sex cord stromal tumor, Paget's disease, pancreatic cancer, papillary cancer, paranasal sinus cancer, parathyroid cancer, pelvic cancer, penile cancer, peripheral nerve cancer, peritoneal cancer, pharyngeal cancer, pheochromocytoma, pilocytic astrocytoma, pineal region tumor, pineoblastoma, pituitary cancer, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, renal cell carcinoma, renal pelvis cancer, rhabdomyosarcoma, salivary gland cancer, soft tissue sarcoma, osteosarcoma, sarcoma, sinus cancer, skin cancer, small cell lung cancer (SCLC), small intestine cancer, spinal column cancer, spinal cord cancer, squamous cell carcinoma, gastric cancer, synovial sarcoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma / thymic carcinoma, thyroid cancer, tongue cancer, tonsil cancer, transitional cell carcinoma, tubular carcinoma, tubular carcinoma, undiagnosed cancer, ureter cancer , urethral cancer, uterine adenocarcinoma, uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, T-cell acute lymphoblastic leukemia (T-ALL), T-cell lymphoblastic lymphoma (T-LL), peripheral T-cell lymphoma, adult T-cell leukemia, precursor B-cell ALL, precursor B-cell lymphoma, large B-cell lymphoma, Burkitt lymphoma, B-cell ALL, Philadelphia chromosome positive ALL, Philadelphia chromosome positive CML, juvenile myelomonocytic leukemia (JMML), acute promyelocytic leukemia (subtype of AML), large granular lymphocyte leukemia, adult T-cell chronic leukemia, diffuse large B-cell lymphoma,Follicular lymphoma; mucosa-associated lymphoid tissue lymphoma (MALT), small cell lymphoma, mediastinal large B-cell lymphoma, nodal marginal zone B-cell lymphoma (NMZL); splenic marginal zone lymphoma (SMZL); intravascular large B-cell lymphoma; primary effusion lymphoma; or lymphomatoid granulomatosis; B-cell prolymphocytic leukemia; splenic lymphoma / leukemia, unclassifiable, splenic diffuse red pulp small B-cell lymphoma; lymphoplasmacytic lymphoma; heavy chain disease, e.g., alpha heavy chain disease, gamma heavy chain disease, mu heavy chain disease, plasma cell myeloma, solitary plasmacytoma of bone; extraosseous plasmacytoma; primary cutaneous follicular center lymphoma, T cell These include: follicular / histiocytic-rich large B-cell lymphoma, chronic inflammation-associated DLBCL; age-related Epstein-Barr virus (EBV)-positive DLBCL; primary mediastinal (thymic) large B-cell lymphoma, primary cutaneous DLBCL, leg type, ALK-positive large B-cell lymphoma, plasmablastic lymphoma; large B-cell lymphoma arising in HHV8-associated multicentric Castleman disease; B-cell lymphoma with features intermediate between diffuse large B-cell lymphoma, unclassified, or B-cell lymphoma with features intermediate between diffuse large B-cell lymphoma and classical Hodgkin lymphoma, unclassified.

[0019] In some embodiments, the cancer is a malignant tumor, a rhabdoid tumor, a CD8+ T-cell lymphoma, an endometrial cancer, an ovarian cancer, a bladder cancer, a gastric cancer, a pancreatic cancer, an esophageal cancer, a prostate cancer, a renal cell carcinoma, a melanoma, a colorectal cancer, a sarcoma (e.g., a soft tissue sarcoma, a synovial sarcoma, an Ewing's sarcoma, an osteosarcoma, an rhabdomyosarcoma, an adult fibrosarcoma, an alveolar soft part sarcoma, angiosarcoma, a clear cell sarcoma, a desmoplastic small round cell tumor, an epithelioid sarcoma, an fibromyxoid sarcoma, an gastrointestinal stromal tumor, a Kaposi's sarcoma, a liposarcoma, a leiomyosarcoma, an malignant mesenchymal tumor, an malignant peripheral nerve sheath tumor, an myxofibrosarcoma, an angio ... In some embodiments, the cancer is malignant tumor, rhabdoid tumor, CD8+T cell lymphoma, endometrial cancer, ovarian cancer, bladder cancer, gastric cancer, pancreatic cancer, esophageal cancer, prostate cancer, renal cell carcinoma, melanoma, or colorectal cancer.In some embodiments, the cancer is sarcoma (e.g., synovial sarcoma or Ewing's sarcoma), non-small cell lung cancer (e.g., squamous cell or adenocarcinoma), gastric cancer, or breast cancer.In some embodiments, the cancer is sarcoma (e.g., synovial sarcoma or Ewing's sarcoma).In some embodiments, the sarcoma is synovial sarcoma.

[0020] In some embodiments of any of the aforementioned methods, the cancer is prostate cancer.In some embodiments of any of the aforementioned methods, the cancer is prostate cancer. In some embodiments of any of the aforementioned methods, the cancer is a BRCA mutated cancer.

[0021] In another aspect, the disclosure relates to a method of treating a viral infection in a patient in need thereof, comprising administering to the subject an effective amount of any of the aforementioned compounds, or a pharma- ceutically acceptable salt thereof, or any of the aforementioned pharmaceutical compositions. In some embodiments, the viral infection is a retrovirus from the family Retroviridae, such as lentiviruses (e.g., human immunodeficiency virus (HIV) and delta retroviruses (e.g., human T-cell leukemia virus I (HTLV-I), human T-cell leukemia virus II (HTLV-II)); Hepadnaviridae (e.g., hepatitis B virus (HBV)), Flaviviridae (e.g., hepatitis C virus (HCV)), Adenoviridae (e.g., human adenovirus), Herpesviridae (e.g., human cytomegalovirus (HCMV), Epstein-Barr virus, herpes simplex virus 1 (HSV-1), herpes simplex virus 2 (HSV-2), human herpes virus 6 (HHV-6), herpes vitus virus (Herpes vitus K), herpes simplex virus (HSV-K ... * , CMV, Varicella-Zoster Virus), Papillomaviridae (e.g., human papillomavirus (HPV, HPVE1)), Parvoviridae (e.g., parvovirus B19), Polyomaviridae (e.g., JC virus and BK virus), Paramyxoviridae (e.g., measles virus), and Togaviridae (e.g., rubella virus).

[0022] In another embodiment of any of the preceding methods, the method further comprises administering to the subject an additional anti-cancer therapy (eg, chemotherapy or a cytotoxic agent or radiation therapy). In some embodiments, the additional anti-cancer therapy is a PARP inhibitor (e.g., niraparib, olaparib, rucaparib, talazoparib, veliparib, pamiparib, CK-102, or E7016). In certain embodiments, the additional anti-cancer therapy is a chemotherapy or cytotoxic agent (e.g., doxorubicin or ifosfamide), a differentiation inducer (e.g., retinoic acid, vitamin D, cytokines), a hormonal agent, an immunological agent, or an anti-angiogenic agent. Chemotherapy and cytotoxic agents include, but are not limited to, alkylating agents, cytotoxic antibiotics, antimetabolites, vinca alkaloids, etoposide, and others (e.g., paclitaxel, taxol, docetaxel, taxotere, cis-platinum). A list of additional compounds with anti-cancer activity 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.

[0023] In certain embodiments, the compound of the present invention and the additional anti-cancer therapy and any of the aforementioned compounds or pharmaceutical compositions 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., 12, 6, 3, 2, or 1 hours; or simultaneously), each in amounts that, together, are effective to treat the subject.

[0024] chemical terms The compounds described herein 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 diastereoisomers, mixtures of diastereoisomers, diastereoisomeric racemates, or mixtures of diastereoisomeric racemates. Optically active forms can be obtained, for example, by resolution of racemates, 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 a pair of stereoisomers whose mirror images are not superimposable, most commonly because they contain asymmetrically substituted carbon atoms that act as chiral centers. "Enantiomer" means one of a pair of molecules that are mirror images of each other and are not superimposable. Diastereomers are stereoisomers that are not related to mirror images, most commonly because they contain two or more asymmetrically substituted carbon atoms and represent the arrangement of substituents around one or more chiral carbon atoms. Enantiomers of a compound can be prepared by separating the 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 the enantiomers of the compounds described herein from a racemic mixture can be readily determined by one of ordinary skill in the art. "Racemate" or "racemic mixture" refers to a compound containing two enantiomers, such mixtures do not exhibit optical activity, i.e., do not rotate the plane of polarized light. "Geometric isomer" refers to isomers that differ in the orientation of substituent atoms in relation to a carbon-carbon double bond, a cycloalkyl ring, or a bridged bicyclic ring system. Atoms (other than H) on each side of a carbon-carbon double bond can be in the E configuration (substituents are on opposite sides of the carbon-carbon double bond) or the Z configuration (substituents are oriented on the same side). "R", "S", "S" are examples of isomers that are different in the orientation of substituent atoms in relation to a carbon-carbon double bond, a cycloalkyl ring, or a bridged bicyclic ring system. * ", "R *", "E", "Z", "cis", and "trans" indicate configurations relative to the core molecule. Some of the disclosed compounds may exist in the form of atropisomers. Atropisomers are stereoisomers resulting from hindered rotation around a single bond, where the steric strain barrier to rotation is high enough to allow for the isolation of the conformers. The compounds described herein can be prepared as individual isomers by isomer-specific synthesis or isolated from an isomeric mixture. Traditional resolution techniques include using optically active acids to form salts of the free bases of each isomer of the isomeric pair (followed by fractional crystallization and regeneration of the free bases), using optically active amines to form salts of the acid forms of each isomer of the isomeric pair (followed by fractional crystallization and regeneration of the free acids), using optically pure acids, amines, or alcohols to form esters or amides of each of the isomers of the isomeric pair (followed by chromatographic separation and removal of the chiral auxiliary), or isomeric mixtures of either the starting materials or the final products using a variety of well-known chromatographic methods. When the stereochemistry of the disclosed compounds is named or depicted by structure, the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight pure relative to the other stereoisomer. When a single enantiomer is named or depicted by structure, the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight optically pure. When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight pure. Percent optical purity is the ratio of the weight of the enantiomer or the weight of the enantiomer plus its optical isomer to the weight of the enantiomer. 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 depicted by structure, the named or depicted 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 depicted by structure, the depicted 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 depicted by structure, the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by mole fraction. The percent purity by mole fraction is the ratio of moles of enantiomer or moles of enantiomer plus moles of its optical isomer. Similarly, percent purity by mole fraction is the ratio of moles of a diastereomer, or moles of a diastereomer plus moles of its isomer. When a disclosed compound is named or depicted by a structure without showing stereochemistry and the compound has at least one chiral center, the name or structure is to be understood to include either an enantiomer of the compound free of the corresponding optical isomer, a racemic mixture of the compound, or a mixture enriched in one enantiomer compared to its corresponding optical isomer. When a disclosed compound is named or depicted by a structure without showing stereochemistry and the compound has two or more chiral centers, the name or structure is to be understood to include a diastereomer free of the other diastereomer, some diastereomers free of other diastereomeric pairs, mixtures of diastereomers, mixtures of diastereomeric pairs, mixtures of diastereomers enriched in one diastereomer over the other diastereomer(s), or mixtures of diastereomers enriched in one or more diastereomers over the other diastereomers. The present invention encompasses all of these forms.

[0025] The compounds of the present disclosure also include all isotopes of atoms occurring in the intermediate or final compounds. "Isotopes" refers to atoms having the same atomic number but different mass numbers due to different numbers of neutrons in the nuclei. For example, isotopes of hydrogen include tritium and deuterium.

[0026] 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) may be useful in compound or substrate tissue distribution assays. 3 H) and carbon-14 (i.e. 14 C) isotopes can be useful in terms of their ease of preparation and detectability. Additionally, deuterium (i.e., 2 Substitution with heavier isotopes, such as H, may provide certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements). 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, 11C 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 using isotopically labeled reagents instead of non-isotopically labeled reagents.

[0027] As is known in the art, many chemical entities can take on a variety of different solid forms, such as, for example, amorphous or crystalline forms (e.g., polymorphs, hydrates, solvates). In some embodiments, the compounds of the present invention can be utilized in any such form, including any solid form. In some embodiments, the compounds described or depicted herein can be provided or utilized in the form of a hydrate or solvate.

[0028] 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 can also be used. The 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.

[0029] 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 "including" and "comprising" may be understood to encompass the individual component or step, whether presented by itself or together with one or more additional components or steps.

[0030] 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.

[0031] As used herein, the term "administration" refers to administering a composition (e.g., a compound or a preparation comprising a compound described herein) to a subject or system. Administration to an animal subject (e.g., a human) can be by any suitable route. For example, in some embodiments, administration can be bronchial (including by bronchial instillation), buccal, enteral, intradermal, intraarterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intratumoral, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including intratracheal instillation), intradermal, intravaginal, and intravitreal.

[0032] As used herein, the term "adult soft tissue sarcoma" refers to sarcomas that typically arise in the soft tissues of the body of adolescent and adult subjects (e.g., subjects who are at least 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 years of age). Non-limiting examples of adult soft tissue sarcomas include, but are not limited to, synovial sarcoma, fibrosarcoma, malignant fibrous histiocytoma, dermatofibrosarcoma, liposarcoma, leiomyosarcoma, angiosarcoma, Kaposi's sarcoma, lymphangiosarcoma, malignant peripheral nerve sheath tumor / neurofibrosarcoma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, extraskeletal myxoid chondrosarcoma, and extraskeletal mesenchymal.

[0033] As used herein, the term "BAF complex" refers to the BRG1 or HRBM-associated factor complex in human cells. As used herein, the term "BAF complex-associated disorder" refers to a disorder caused by or affected by the level and / or activity of the BAF complex.

[0034] As used herein, the terms "GBAF complex" and "GBAF" refer to the SWI / SNF ATPase chromatin remodeling complex in human cells. GBAF complex subunits may include, but are not limited to, ACTB, ACTL6A, ACTL6B, BICRA, BICRAL, BRD9, SMARCA2, SMARCA4, SMARCC1, SMARCD1, SMARCD2, SMARCD3, and SS18. The term "cancer" refers to a condition caused by the proliferation of malignant neoplastic cells, such as tumors, neoplasms, carcinomas, sarcomas, leukemias, and lymphomas.

[0035] As used herein, the term "BRD9" refers to bromodomain-containing protein 9, a component of the BAF (BRG1 or BRM-associated factor) complex, a SWI / SNF ATPase chromatin remodeling complex, and a member of family IV of bromodomain-containing proteins. BRD9 is encoded by the BRD9 gene, the nucleic acid sequence of which is set forth in SEQ ID NO:1. The term "BRD9" also refers to naturally occurring variants of the wild-type BRD9 protein, such as proteins having at least 85% sequence identity (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9% identity, or more) to the amino acid sequence of wild-type BRD9, as set forth in SEQ ID NO:2.

[0036] As used herein, the term "BRD9-associated disorder" refers to a disorder caused or affected by the level and / or activity of BRD9. The term "cancer" refers to a condition caused by the proliferation of malignant neoplastic cells, such as tumors, neoplasms, carcinomas, sarcomas, leukemias, and lymphomas.

[0037] 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 is simultaneous or parallel, and the agents may be co-formulated. In some embodiments, the two or more agents are not co-formulated, but are administered sequentially as part of a prescribed regimen. In some embodiments, the administration of the two or more agents or treatments in combination 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 be 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.

[0038] "Compounds of the invention" and similar terms, as used herein, refer to compounds useful for treating BAF-related disorders (e.g., cancer or infectious diseases) described herein, such as, for example, Compound D1, Compound S-D1, Compound R-D1, or Compound D2, as well as salts (e.g., pharma-ceutically acceptable salts), solvates, hydrates, stereoisomers (including atropisomers), and tautomers thereof, whether or not expressly indicated. Those skilled in the art will understand that certain compounds described herein can exist in one or more different isomeric (e.g., stereoisomers, geometric isomers, atropisomers, and tautomers) or isotopic (e.g., one or more atoms are replaced with different isotopes of the atom, e.g., hydrogen is replaced with deuterium) forms. Unless otherwise specified or clear from the context, the depicted structures can be understood to represent any such isomeric or isotopic forms, individually or in combination. Compounds described herein can be asymmetric (e.g., have one or more stereocenters). All stereoisomers, including enantiomers and diastereomers, are intended unless otherwise specified. Compounds of the present disclosure that contain asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods on how to prepare optically active forms from optically active starting materials are known in the art, for example, by resolution of racemic mixtures or by stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, and the like can also be present in the compounds described herein, and all such stable isomers are contemplated in the present disclosure. Cis and trans geometric isomers of the compounds of the present disclosure are described and can be isolated as a mixture of isomers or as separated isomeric forms. In some embodiments, one or more compounds depicted herein can exist in different tautomeric forms. Reference to such compounds encompasses all such tautomeric forms, unless expressly excluded, as is clear from the context. In some embodiments, tautomeric forms result from the exchange of a single bond with an adjacent double bond and the concomitant migration of a proton.In certain embodiments, tautomeric forms can be prototropic tautomers, which are isomeric protonation states with the same empirical formula and total charge as the reference form. Examples of moieties with prototropic tautomeric forms are ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, amide-imidic acid pairs, enamine-imine pairs, and cyclic forms in which protons can occupy more than one position of a heterocyclic ring system, such as 1H- and 3H-imidazole, 1H-, 2H-, and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. In some embodiments, tautomeric forms can be in equilibrium or sterically fixed in one form by appropriate substitution. In certain embodiments, tautomeric forms are produced by acetal interconversion.

[0039] As used herein, the term "degrading agent" refers to a small molecule compound that contains a degrading moiety, where the compound interacts with a protein (e.g., BRD9) in a manner that results in degradation of the protein, e.g., binding of the compound results in at least a 5% decrease in the levels of the protein, e.g., in a cell or subject.

[0040] As used herein, the term "degradation moiety" refers to a moiety whose binding results in the degradation of a protein, e.g., BRD9. In one example, the moiety binds to a protease or ubiquitin ligase that metabolizes a protein, e.g., BRD9.

[0041] "Determining the level of a protein" means detecting the protein or RNA encoding the protein, 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 obtaining a physical entity or value from another party or source (e.g., a third-party testing 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 mRNA levels are known in the art.

[0042] As used herein, the terms "effective amount," "therapeutically effective amount," and "sufficient amount" of an agent that reduces the level and / or activity of BRD9 described herein (e.g., in a cell or a subject) refer to an amount sufficient to bring about a beneficial or desired result, e.g., a clinical result, when administered to a subject, including a human, and thus "effective amount" or its synonyms will depend on the context in which it is being applied. For example, in the context of treating cancer, it is the amount of an agent that reduces the level and / or activity of BRD9 sufficient to achieve a therapeutic response compared to the response obtained in the absence of administration of the agent that reduces the level and / or activity of BRD9. The amount of a given agent that reduces the level and / or activity of BRD9 described herein that would correspond to such an amount will vary depending on various factors, such as the given agent, pharmaceutical formulation, route of administration, type of disease or disorder, personality of the subject (e.g., age, sex, and / or weight) or host being treated, but can nevertheless be routinely determined by one of skill in the art. Also, as used herein, a "therapeutically effective amount" of an agent that reduces the level and / or activity of BRD9 of the present disclosure is an amount that produces a beneficial or desired result in a subject compared to a control. As defined herein, a therapeutically effective amount of an agent that reduces the level and / or activity of BRD9 of the present disclosure can be easily determined by a person skilled in the art by routine methods known in the art. Dosage regimens can be adjusted to provide an optimal therapeutic response.

[0043] As used herein, the term "inhibitor" refers to any agent that reduces the level and / or activity of a protein (e.g., BRD9). Non-limiting examples of inhibitors include small molecule inhibitors, degraders, antibodies, enzymes, or polynucleotides (e.g., siRNAs).

[0044] "Level" refers to the level of a protein or an mRNA encoding a protein, as compared to a reference. The reference can be any useful reference as defined herein. A "reduced level" or "increased level" of a protein refers to a reduction or increase in protein level compared to a reference (e.g., 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 reduction or increase; as compared to a reference, (e.g., greater than about 10%, about 15%, about 20%, about 50%, about 75%, about 100%, or about 200% reduction or increase; 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 reduction or increase; greater than 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 increase). Protein levels can be expressed as mass / volume (e.g., g / dL, mg / mL, μg / mL, ng / mL, etc.) or as a percentage of total protein or mRNA in the sample.

[0045] "Modulating the activity of the BAF complex" means modifying the level of activity associated with the BAF complex (e.g., GBAF) or the associated downstream effect. The activity level of the BAF complex can be measured using any method known in the art (e.g., the method described in Kadoch et al, Cell 153:71-85 (2013)), which is incorporated herein by reference.

[0046] "Percent (%) sequence identity" with respect to a reference polynucleotide or polypeptide sequence is defined as the percentage of nucleic acids or amino acids in a candidate sequence that are identical to the nucleic acids or amino acids in the reference polynucleotide or polypeptide sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity. Alignment for purposes of determining percent nucleic acid or amino acid sequence identity can be accomplished in a variety of ways that are within the capabilities of those skilled in the art, for example, using publicly available computer software such as BLAST, BLAST-2, or Megalign software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms necessary to achieve maximum alignment over the entire length of the sequences being compared. For example, percent sequence identity values ​​can be generated using the sequence comparison computer program BLAST. By way of illustration, the percent sequence identity of a given nucleic acid or amino acid sequence A to, with, or relative to a given nucleic acid or amino acid sequence B (which can alternatively be expressed as a given nucleic acid or amino acid sequence A having a certain percent sequence identity to, with, or relative to a given nucleic acid or amino acid sequence B) is calculated as follows: Multiply by 100(fraction X / Y) where X is the number of nucleotides or amino acids scored by a sequence alignment program (e.g., BLAST) as identical matches in that program's alignment of A and B, and Y is the total number of nucleic acids in B. It will be understood that if the length of nucleic acid or amino acid sequence A is not equal to the length of nucleic acid or amino acid sequence B, then the percent sequence identity of A to B will not equal the percent sequence identity of B to A.

[0047] "Pharmaceutically acceptable excipient" as used herein refers to any component other than the compounds described herein (e.g., a vehicle capable of suspending or dissolving active compounds) that has the property of being substantially non-toxic and non-inflammatory in patients.Excipients may include, for example, anti-adherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colorings), emollients, emulsifiers, fillers (diluents), film-forming or coating agents, flavors, fragrances, lubricants (glidants), lubricants, preservatives, printing inks, adsorbents, suspending or dispersing agents, sweeteners, and water for hydration.

[0048] As used herein, the term "pharmaceutically acceptable salt" refers to any pharmaceutically acceptable salt of any compound described herein. For example, any pharmaceutically acceptable salt of any compound described herein includes salts that are suitable for use in contact with human and animal tissues within the scope of sound medical judgment, without toxicity, irritation, allergic response, etc., 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 can be prepared separately by reacting the free base group with a suitable organic acid. The compounds described herein 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 in the case of the acidic form of the compounds described herein.In many cases, 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, as well as methods for preparing 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.

[0049] The term "pharmaceutical composition" as used herein refers to a composition containing a compound described herein that is formulated with a pharmaceutically acceptable excipient and manufactured or sold with the approval of a government regulatory agency as part of a therapeutic regimen for the treatment of a disease in a mammal.The pharmaceutical composition can be formulated, for example, in unit dosage form for oral administration (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 that is free of particulate embolic material and in a solvent system suitable for intravenous use); or in any other pharmaceutically acceptable formulation.

[0050] "Reducing the activity of BRD9" means reducing the level of activity associated with BRD9 or the associated downstream effect. A non-limiting example of inhibiting the activity of BRD9 is reducing the level of BAF complex (e.g., GBAF) in cells. The activity level of BRD9 can be measured using any method known in the art. In some embodiments, the agent that reduces the activity of BRD9 is a small molecule BRD9 inhibitor. In some embodiments, the agent that reduces the activity of BRD9 is a small molecule BRD9 degrader.

[0051] "Reducing the level of BRD9" means reducing the level of BRD9 in a cell or a subject. The level of BRD9 can be measured using any method known in the art.

[0052] By "reference" is meant any useful reference used to compare protein or mRNA levels. A reference can be any sample, standard, standard curve, or level used for comparison purposes. A reference 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) that does not have a disease; a sample from a subject diagnosed with a disease but not yet treated with a compound described herein; a sample from a subject treated with a compound described herein; or a sample of a known normal concentration of purified protein (e.g., any of those described herein). By "reference standard or level" is meant a value or number obtained from a reference sample. A "normal control value" is a predefined value that indicates a non-disease state, e.g., a value that would be expected in a healthy control subject. Typically, a normal control value is expressed as a range ("X to Y"), a high threshold value ("below X"), or a low threshold value ("above X"). A subject having a measured 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 treated with a compound described herein. 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 levels within the normal reference range, e.g., any of those described herein, can also be used as a reference.

[0053] As used herein, the term "subject" refers to any organism to which a composition according to the invention may be administered, e.g., 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 that is seeking or in need of treatment, is in need of treatment, is undergoing treatment, may be undergoing treatment, or is receiving the care of a skilled professional for a particular disease or condition.

[0054] As used herein, the term "SS18-SSX fusion protein associated disorder" refers to a disorder caused or affected by the level and / or activity of a SS18-SSX fusion protein.

[0055] As used herein, the terms "treat", "treated" or "treating" refer to both therapeutic treatments and prophylactic or preventative measures aimed at preventing or slowing (alleviating) an undesirable physiological condition, disorder, or disease, or obtaining a beneficial or desired clinical outcome. Beneficial or desired outcomes may include, but are not limited to, alleviation of symptoms; reduction in the extent of a condition, disorder, or disease; a stable (i.e., non-worsening) state of a condition, disorder, or disease; a delay or slowing of the onset of progression of a condition, disorder, or disease; an improvement or remission (partial or total) of a condition, disorder, or pathology, whether detectable or undetectable; an improvement in at least one measurable physical parameter (not necessarily discernible by the patient); or an improvement or amelioration of a condition, disorder, or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival compared to the expected survival if not receiving treatment.

[0056] As used herein, the terms "variant" and "derivative" are used interchangeably and refer to naturally occurring, synthetic, and semi-synthetic analogs of the compounds, peptides, proteins, or other substances described herein. Variants or derivatives of the compounds, peptides, proteins, or other substances described herein may retain or improve the biological activity of the original material.

[0057] The details of one or more embodiments of the invention are set forth in the detailed description below. Other features, objects, and advantages of the invention will become apparent from the description and the claims. [Brief description of the drawings]

[0058] [Figure 1] 1 is an image showing dose-dependent depletion of BRD9 levels in a synovial sarcoma cell line (SYO1) in the presence of a BRD9 degrader. [Diagram 2] 13 is an image showing sustained suppression of BRD9 levels in a synovial sarcoma cell line (SYO1) in the presence of a BRD9 degrader for 72 hours. [Diagram 3] 13 is an image showing sustained suppression of BRD9 levels in two cell lines (293T and SYO1) in the presence of a BRD9 degrader over a period of 5 days. [Figure 4] 13 is an image showing sustained suppression of BRD9 levels in synovial sarcoma cell lines (SYO1 and Yamato) in the presence of a BRD9 degrader over a 7-day period compared to levels in cells treated with a CRISPR reagent. [Diagram 5] These are images showing the effect on cell proliferation of six cell lines (SYO1, Yamato, A549, HS-SY-II, ASKA, and 293T) in the presence of BRD9 degraders and BRD9 inhibitors. [Figure 6] 13 is an image showing the effect on cell proliferation of two cell lines (SYO1 and G401) in the presence of a BRD9 degrader. [Figure 7]13 is an image showing the effect on cell proliferation of three synovial sarcoma cell lines (SYO1, HS-SY-II, and ASKA) in the presence of a BRD9 degrader, a BRD9 binder, and an E3 ligase binder. [Figure 8] 13 is an image showing the effect on cell proliferation of three non-synovial sarcoma cell lines (RD, HCT116, and Calu6) in the presence of a BRD9 degrader, a BRD9 binder, and an E3 ligase binder. [Figure 9] 1 is a graph showing the percentage of SYO1 in various cell cycle phases after 8 or 13 days of treatment with DMSO, Compound 1 at 200 nM, or Compound 1 at 1 μM. [Figure 10] A series of contour plots showing the percentage of SYO1 cells in various cell cycle phases after 8 days of treatment with DMSO, Compound 1 at 200 nM, Compound 1 at 1 μM, or Lenalidomide at 200 nM. The values ​​corresponding to each contour plot are found in the table below. [Figure 11] 1 is a series of contour plots showing the percentage of SYO1 cells in various cell cycle phases after treatment with DMSO, Compound 1 at 200 nM, Compound 1 at 1 μM, or Lenalidomide at 200 nM for 13 days. The values ​​corresponding to each contour plot are found in the table below. [Figure 12] 1 is a series of contour plots showing the percentage of early and late apoptotic SYO1 cells after 8 days of treatment with DMSO, Compound 1 at 200 nM, Compound 1 at 1 μM, or Lenalidomide at 200 nM. The values ​​corresponding to each contour plot are found in the table below. [Figure 13] 1 is a graph showing proteins present in a BAF complex containing a SS18-SSX fusion protein. [Figure 14] 1 is a graph showing the efficacy of compound D1 in a SOY-1 xenograft mouse model. Treatment with compound D1 resulted in tumor growth inhibition. [Figure 15]1 is an image of a Western blot showing BRD9 detection in control and treated groups (compound D1). Treatment with compound D1 resulted in BRD9 inhibition. [Figure 16] 13 is an image of a Western blot showing BRD9 detection in SYO-1 cells treated with DMSO, enantiomer 1, or racemic compound D1 for 1 or 6 hours. [Figure 17] 13 is an image of a Western blot showing BRD9 detection in SYO-1 cells treated with DMSO, enantiomer 2, or racemic compound D1 for 1 or 6 hours. [Figure 18] FIG. 18 is a graph showing dose-response curves fitted to the BRD9 binding intensity data points from the Western blot images shown in FIGS. 16 and 17. [Figure 19] 1 is an image of a Western blot showing BRD9 detection in SYO-1 cells treated with Enantiomer 1, Enantiomer 2, or racemic compound D1 for 24 hours. [Figure 20] 1 is an image of a Western blot showing BRD9 detection in ASKA cell control and ASKA cells treated with enantiomer 1 or racemic compound D1 for 0.5 or 2 hours. [Figure 21] 13 is an image of a Western blot showing BRD9 detection in ASKA cell control and ASKA cells treated with enantiomer 2 or racemic compound D1 for 0.5 or 2 hours. [Figure 22] FIG. 22 is a graph showing dose-response curves fitted to the BRD9 binding intensity data points from the Western blot images shown in FIGS. 20 and 21. [Figure 23] 1 is a series of images showing Western blots for BRD9 detection in SYO-1 Zenograft models treated with Enantiomer 1, Enantiomer 2, or racemic compound D1. [Figure 24] FIG. 24 is a bar graph quantifying the BRD9 level changes observed in the Western blots shown in FIG. 23. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0059] The present disclosure relates to compositions and methods useful for treating BAF-associated disorders, such as cancer and infectious diseases. The present disclosure further relates to compositions and methods useful for inhibiting the level and / or activity of BRD9, e.g., treating disorders, such as cancer (e.g., sarcoma) and infectious diseases (e.g., viral infections), e.g., in a subject in need thereof.

[0060] compound The compounds described herein reduce the level of BRD9-related activity or associated downstream effects or reduce the level of BRD9 in a cell or subject. Exemplary compounds described herein, or pharma- ceutically acceptable salts thereof, have the structure of compound D1, S-D1, R-D1, or D2 in Table 1.

[0061] Medical Use The compounds described herein are useful in the methods of the invention and, without being bound by theory, are believed to exert their desired effects through their ability to modulate the levels, status, and / or activity of the BAF complex, for example, by inhibiting the activity or levels of the BRD9 protein in cells within the BAF complex in a mammal.

[0062] Certain aspects of the invention relate to methods of treating a BRD9-associated disorder, such as cancer, in a subject in need thereof. In some embodiments, the compound is administered in an amount and for a time effective to result in one (or more, e.g., two or more, three or more, four or more) of the following: (a) a decrease in tumor size, (b) a decrease in tumor growth rate, (c) an increase in tumor cell death, (d) a decrease in tumor progression, (e) a decrease in the number of metastases, (f) a decrease in the rate of metastases, (g) a reduction in tumor recurrence, (h) an increase in the subject's survival time, and (i) an increase in the subject's progression-free survival time.

[0063] Treatment of 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) compared to the tumor size before treatment. Tumor size can be measured by any reproducible measurement means. For example, tumor size can be measured as the diameter of the tumor.

[0064] Treatment of cancer can further result in the reduction of tumor number.For example, after treatment, tumor number is reduced by 5% or more (for example, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) compared to the number before treatment.Tumor number can be measured by any reproducible measurement means, for example, tumor number can be measured by counting tumors visible to the naked eye or at a certain magnification (for example, 2x, 3x, 4x, 5x, 10x, or 50x).

[0065] Treatment of cancer can reduce 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 visible to the naked eye or at a certain magnification (e.g., 2x, 10x, or 50x).

[0066] Treatment of 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 extension of the average survival time of a population can be measured, for example, by calculating the average survival time of a population after starting treatment with a compound described herein. The extension of the average survival time of a population can also be measured, for example, by calculating the average survival time of a population after completing the first round of treatment with a pharma- ceutically acceptable salt of a compound described herein.

[0067] Cancer treatment can 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 can be measured by any reproducible means, for example, by calculating the average number of disease-related deaths per unit time for a population after starting treatment with a pharma- ceutically acceptable salt of a compound described herein. The reduction in the mortality rate of a population can be measured, for example, by calculating the average number of disease-related deaths per unit time for a population after completing the first round of treatment with a pharma- ceutically acceptable salt of a compound described herein.

[0068] Combination therapy The method of the present invention can be used alone or in combination with additional therapeutic agents, such as additional therapeutic agents that treat cancer or symptoms related thereto, or in combination with other types of treatments for treating cancer. In combination therapy, 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 is that which produces a therapeutic effect.

[0069] In some embodiments, the second therapeutic agent is a chemotherapeutic agent (e.g., a cytotoxic agent or other chemical 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, adrenocortical suppressants, adrenocortical steroids, progestins, estrogens, antiestrogens, androgens, antiandrogens, and gonadotropin-releasing hormone analogs. Also included are 5-fluorouracil (5-FU), leucovorin (LV), irinotecan, oxaliplatin, capecitabine, paclitaxel, and doxetaxel. Non-limiting examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclosphosphamide; alkylsulfonates, e.g., busulfan, improsulfan, and piposulfan; aziridines, e.g., benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines, e.g., altretamine, triethylenemelamine, triethylenephosphoramide, triethylylenethiophosphoramide, and trimethylolomelanin; acetogenins (especially bullatacin and bullatacinone); camptothecins (including the synthetic analog topotecan); bryostatin; kallistatin; CC-1 065 (including its synthetic analogs adozelesin, carzelesin and bizelesin); cryptophycins (especially cryptophycin 1 and cryptophycin 8); dolastatins; duocarmycins (including synthetic analogs, KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; spongistatins; nitrogen mustards such as chlorambucil, chlornaphazine, colofosfamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard;nitrosoureas, e.g., carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine; antibiotics, e.g., enediyne antibiotics (e.g., the calicheamicins, especially calicheamicin gamma and calicheamicin omega II (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, carabicin, caminomycin, carzinov; doxorubicin, 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, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycin, riboflavin ... antimetabolites such as mycin, peplomycin, potfilomycin, puromycin, keramycin, rhodrubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin, methotrexate, and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, and trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine; ancitabine Pyrimidine analogues such as cyclosporine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, and floxuridine; androgens such as calsterone, dromostanolone propionate, epithiostanol, mepitiostane, and testolactone; antiadrenal agents such as aminoglutethimide, mitotane, and trilostane; folic acid replacements 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, mopidamnol, nitraelin, pentostatin; Fenameth; Pirarubicin; Rosoxantrone; Podophyllic acid; 2-Ethylhydrazide; Procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, OR), 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® (chromophore free), albumin engineered nanoparticle formulations of paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.), and TAXOTERE® docetaxel (Rhone-Poulenc Rorer, Antony, France); chloranbucil; GEMZAR® gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum coordination complexes such as cisplatin, oxaliplatin, and 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 include RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; capecitabine; and pharma- ceutically acceptable salts, acids, and derivatives of any of the above. Two or more chemotherapeutic agents can be used in cocktails to be administered in combination with the 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., Proc. Am. Soc. Clin. Oncol. 18:233a (1999), and Douillard et al., Lancet 355(9209):1041-1047 (2000).

[0070] In some embodiments, the second therapeutic agent is a DNA damaging agent (eg, platinum-based antineoplastic agents, topoisomerase inhibitors, PARP inhibitors, alkylating antineoplastic agents, and ionizing radiation).

[0071] Examples of platinum-based anti-neoplastic agents that may be used as second therapeutic agents in the compositions and methods of the present invention are cisplatin, carboplatin, oxaliplatin, dicycloplatin, eptaplatin, lobaplatin, miriplatin, nedaplatin, triplatin tetranitrate, phenanthrilplatin, picoplatin, and satraplatin. In some embodiments, the second therapeutic agent is cisplatin and the cancer being treated is testicular cancer, ovarian cancer, or bladder cancer (e.g., advanced bladder cancer). In some embodiments, the second therapeutic agent is carboplatin and the cancer being treated is ovarian cancer, lung cancer, head and neck cancer, brain cancer, or neuroblastoma. In some embodiments, the second therapeutic agent is oxaliplatin and the cancer being treated is colorectal cancer. In some embodiments, the second therapeutic agent is dicycloplatin and the cancer being treated is non-small cell lung cancer or prostate cancer. In some embodiments, the second therapeutic agent is eptaplatin and the cancer being treated is gastric cancer. In some embodiments, the second therapeutic agent is lobaplatin and the cancer being treated is breast cancer. In some embodiments, the second therapeutic agent is miriplatin and the cancer being treated is hepatocellular carcinoma. In some embodiments, the second therapeutic agent is nedaplatin and the cancer being treated is nasopharyngeal cancer, esophageal cancer, squamous cell carcinoma, or cervical cancer. In some embodiments, the second therapeutic agent is triplatin tetranitrate and the cancer being treated is lung cancer (e.g., small cell lung cancer) or pancreatic cancer. In some embodiments, the second therapeutic agent is picoplatin and the cancer being treated is lung cancer (e.g., small cell lung cancer), prostate cancer, bladder cancer, or colorectal cancer. In some embodiments, the second therapeutic agent is satrapltin and the cancer being treated is prostate cancer, breast cancer, or lung cancer.

[0072] Examples of topoisomerase inhibitors that can be used as second therapeutic agents in the compositions and methods of the present invention are etoposide, teniposide, doxorubicin, daunorubicin, mitoxantrone, amsacrine, ellipticine, irinotecan, topotecan, camptothecin, and diflomotecan.In some embodiments, the second therapeutic agent is etoposide, and the cancer being treated is lung cancer (e.g., small cell lung cancer) or testicular cancer.In some embodiments, the second therapeutic agent is teniposide, and the cancer being treated is acute lymphoblastic leukemia (e.g., childhood acute lymphoblastic leukemia). In some embodiments, the second therapeutic agent is doxorubicin, and the cancer being treated is acute lymphoblastic leukemia, acute myeloid leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, breast cancer, Wilms' tumor, neuroblastoma, soft tissue sarcoma, osteosarcoma, ovarian cancer, transitional cell carcinoma of the bladder, thyroid cancer, gastric cancer, or bronchogenic lung cancer. In some embodiments, the second therapeutic agent is daunorubicin, and the cancer being treated is acute lymphoblastic leukemia or acute myeloid leukemia. In some embodiments, the second therapeutic agent is mitoxantrone, and the cancer being treated is prostate cancer or acute nonlymphocytic leukemia. In some embodiments, the second therapeutic agent is amsacrine, and the cancer being treated is leukemia (e.g., acute adult leukemia). In some embodiments, the second therapeutic agent is irinotecan, and the cancer being treated is colorectal cancer. In some embodiments, the second therapeutic agent is topotecan and the cancer being treated is lung cancer (e.g., small cell lung cancer). In some embodiments, the second therapeutic agent is diflomotecan and the cancer being treated is lung cancer (e.g., small cell lung cancer).

[0073] Examples of alkylating antineoplastic agents that may be used as second therapeutic agents in the compositions and methods of the present invention are cyclophosphamide, uramustine, melphalan, chlorambucil, ifosfamide, bendamustine, carmustine, lomustine, chlorozotocin, fotemustine, nimustine, ranimustine, busulfan, improsulfan, piposulfan, chlornaphazine, colofosphamide, estramustine, mechlorethamine, mechlorethamine oxide hydrochloride, nobuenbiquine, phenesterine, prednimustine, trofosfamide, procarbazine, altretamine, dacarbazine, mitozolomide, and temozolomide. In some embodiments, the second therapeutic agent is cyclophosphamide and the cancer being treated is Non-Hodgking lymphoma. In some embodiments, the second therapeutic agent is melphalan and the cancer being treated is multiple myeloma, ovarian cancer, or melanoma. In some embodiments, the second therapeutic agent is chlorambucil and the cancer being treated is chronic lymphocytic leukemia, malignant lymphoma (e.g., lymphosarcoma, giant follicular lymphoma, or Hodgkin's lymphoma). In some embodiments, the second therapeutic agent is ifosfamide and the cancer being treated is testicular cancer. In some embodiments, the second therapeutic agent is bendamustine and the cancer being treated is chronic lymphocytic leukemia or non-Hodgkin's lymphoma. In some embodiments, the second therapeutic agent is carmustine and the cancer being treated is brain cancer (e.g., glioblastoma, brainstem glioma, medulloblastoma, astrocytoma, ependymoma, or metastatic brain tumor), multiple myeloma, Hodgkin's disease, or non-Hodgkin's lymphoma. In some embodiments, the second therapeutic agent is lomustine and the cancer being treated is brain cancer or Hodgkin's lymphoma. In some embodiments, the second therapeutic agent is fotemustine and the cancer being treated is melanoma. In some embodiments, the second therapeutic agent is nimustine and the cancer being treated is brain cancer. In some embodiments, the second therapeutic agent is ranimustine and the cancer being treated is chronic myelogenous leukemia or polycythemia vera. In some embodiments, the second therapeutic agent is busulfan and the cancer being treated is chronic myelogenous leukemia.In some embodiments, the second therapeutic agent is improsulfan and the cancer being treated is a sarcoma. In some embodiments, the second therapeutic agent is estramustine and the cancer being treated is prostate cancer (e.g., prostate carcinoma). In some embodiments, the second therapeutic agent is mechlomethamine and the cancer being treated is cutaneous T-cell lymphoma. In some embodiments, the second therapeutic agent is trofosfamide and the cancer being treated is a sarcoma (e.g., soft tissue sarcoma). In some embodiments, the second therapeutic agent is procarbazine and the cancer being treated is Hodgkin's disease. In some embodiments, the second therapeutic agent is altretamine and the cancer being treated is ovarian cancer. In some embodiments, the second therapeutic agent is dacarbazine and the cancer being treated is melanoma, Hodgkin's lymphoma, or sarcoma. In some embodiments, the second therapeutic agent is temozolomide and the cancer being treated is brain cancer (e.g., astrocytoma or glioblastoma) or lung cancer (e.g., small cell lung cancer).

[0074] Examples of PARP inhibitors that can be used as second therapeutic agent in the compositions and methods of the present invention are niraparib, olaparib, rucaparib, talazoparib, veliparib, pamiparib, CK-102, or E7016. Advantageously, the compounds of the present invention and DNA damaging agents can act synergistically to treat cancer. In some embodiments, the second therapeutic agent is niraparib, and the cancer to be treated is ovarian cancer (e.g., BRCA-mutated ovarian cancer), fallopian tube cancer (e.g., BRCA-mutated fallopian tube cancer), or primary peritoneal cancer (e.g., BRCA-mutated primary peritoneal cancer). In some embodiments, the second therapeutic agent is olaparib, and the cancer being treated is lung cancer (e.g., small cell lung cancer), ovarian cancer (e.g., BRCA-mutated ovarian cancer), breast cancer (e.g., BRCA-mutated breast cancer), fallopian tube cancer (e.g., BRCA-mutated fallopian tube cancer), primary peritoneal cancer (e.g., BRCA-mutated primary peritoneal cancer), prostate cancer (e.g., castration-resistant prostate cancer), or pancreatic cancer (e.g., pancreatic adenocarcinoma). In some embodiments, the second therapeutic agent is rucaparib, and the cancer being treated is ovarian cancer (e.g., BRCA-mutated ovarian cancer), fallopian tube cancer (e.g., BRCA-mutated fallopian tube cancer), or primary peritoneal cancer (e.g., BRCA-mutated primary peritoneal cancer). In some embodiments, the second therapeutic agent is talazoparib, and the cancer being treated is breast cancer (e.g., BRCA-mutated breast cancer). In some embodiments, the second therapeutic agent is veliparib and the cancer being treated is lung cancer (e.g., non-small cell lung cancer), melanoma, breast cancer, ovarian cancer, prostate cancer, or brain cancer. In some embodiments, the second therapeutic agent is pamiparib and the cancer being treated is ovarian cancer. In some embodiments, the second therapeutic agent is CK-102 and the cancer being treated is lung cancer (e.g., non-small cell lung cancer). In some embodiments, the second therapeutic agent is E7016 and the cancer being treated is melanoma.

[0075] Without wishing to be bound by theory, the synergistic effect between the compounds of the present invention and DNA damaging agents may be due to the requirement of BRD9 for DNA repair; inhibition of BRD9 increases the sensitivity of cancer (e.g., cancer cells or tissues) to DNA damaging agents.

[0076] In some embodiments, the second therapeutic agent is a JAK inhibitor (e.g., JAK1 inhibitor). Non-limiting examples of JAK inhibitors that can be used as second therapeutic agents in the compositions and methods of the present invention include tofacitinib, ruxolitinib, oclacitinib, baricitinib, peficitinib, fedratinib, upadacitinib, filgotinib, cerduratinib, gandotinib, lestaurtinib, momelotinib, pacritinib, abrocitinib, solcitinib, itacitinib, or SHR0302. Without wishing to be bound by theory, the synergistic effect between the compounds of the present invention and JAK inhibitors may be the inhibitor of SAGA complex for their combined effect of downregulating Foxp3+Treg cells. In some embodiments, the second therapeutic agent is ruxolitinib and the cancer being treated is a myeloproliferative neoplasm (e.g., erythrocytosis or myelofibrosis), ovarian cancer, breast cancer, pancreatic cancer. In some embodiments, the second therapeutic agent is fedratinib and the cancer being treated is a myeloproliferative neoplasm (e.g., myelofibrosis). In some embodiments, the second therapeutic agent is cerduratinib and the cancer being treated is a lymphoma (e.g., peripheral T-cell lymphoma). In some embodiments, the second therapeutic agent is gandotinib and the cancer being treated is a myeloproliferative neoplasm (e.g., erythrocytosis or myelofibrosis). In some embodiments, the second therapeutic agent is lestaurtinib and the cancer being treated is a myeloproliferative neoplasm (e.g., erythrocytosis or myelofibrosis), leukemia (e.g., acute myeloid leukemia), pancreatic cancer, prostate cancer, or neuroblastoma. In some embodiments, the second therapeutic agent is momelotinib and the cancer being treated is a myeloproliferative neoplasm (e.g., erythrocytosis or myelofibrosis) or pancreatic cancer (e.g., pancreatic ductal adenocarcinoma). In some embodiments, the second therapeutic agent is momelotinib and the cancer being treated is a myeloproliferative neoplasm (e.g., erythrocytosis or myelofibrosis). In some embodiments, the second therapeutic agent is momelotinib and the cancer being treated is a myeloproliferative neoplasm (e.g., erythrocytosis or myelofibrosis) or pancreatic cancer (e.g., pancreatic ductal adenocarcinoma).

[0077] In some embodiments, the second therapeutic agent is an inhibitor of the SAGA complex or a component thereof. The SAGA complex inhibitor may be, for example, an inhibitory antibody or small molecule inhibitor of CCDC101, Tada2B, Tada3, Usp22, Tada1, Taf6l, Supt5, Supt20, or a combination thereof. Without wishing to be bound by theory, the synergistic effect between the compound of the present invention and the inhibitor of the SAGA complex may be due to their combined effect of downregulating Foxp3+ Treg cells. In some embodiments, the second therapeutic agent 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 antiangiogenic agent, for example, an anti-VEGF agent, for example, bevacizumab (AVASTIN®). In some embodiments, the biologic is an immunoglobulin-based biologic, e.g., a monoclonal antibody (e.g., a humanized antibody, fully human antibody, Fc fusion protein, or functional fragment thereof) that agonizes a target to stimulate an anti-cancer response or antagonizes an antigen important to cancer.Such agents 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® (tocilizumab);VECTIBIX® (panitumumab);LUCENTIS® (ranibizumab);SOLIRIS® (eculizumab);CIMZIA® (certolizumab) pegol; SIMPONI® (golimumab); ILARIS® (canakinumab); STELARA® (ustekinumab); ARZERRA® (ofatumumab); PROLIA® (denosumab); NUMAX® (motavizumab); ABTHRAX® (raxibacumab); BENLYSTA® (belimumab); YERVOY® (ipilimumab); ADCETRIS® (brentuximab vedotin); PERJETA® (pertuzumab); KADCYLA® (ado-trastuzumab emtansine); and GAZYVA® (obinutuzumab). Antibody-drug conjugates are also included.

[0078] The second agent can be a therapeutic agent that is a non-drug treatment, for example, the second therapeutic agent is radiation therapy, cryotherapy, hyperthermia, and / or surgical removal of tumor tissue. The second agent can 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 can be, for example, a humanized or fully human antibody. 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 a 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 or fusion a protein, e.g., 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 PDL1 (e.g., MPDL3280A / RG7446; MEDI4736; MSB0010718C; BMS936559). 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 AMP224). 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. In some embodiments, the second therapeutic agent is ipilimumab and the cancer being treated is melanoma, renal cancer, lung cancer (e.g., non-small cell lung cancer or small cell lung cancer), or prostate cancer.In some embodiments, the second therapeutic agent is tremelimumab and the cancer being treated is melanoma, mesothelioma, or lung cancer (e.g., non-small cell lung cancer). In some embodiments, the second therapeutic agent is nivolumab and the cancer being treated is melanoma, lung cancer (e.g., non-small cell lung cancer or small cell lung cancer), kidney cancer, Hodgkin's lymphoma, head and neck cancer (e.g., head and neck squamous cell carcinoma), urothelial carcinoma, hepatocellular carcinoma, or colorectal cancer. In some embodiments, the second therapeutic agent is pembrolizumab and the cancer being treated is melanoma, lung cancer (e.g., non-small cell lung cancer or small cell lung cancer), Hodgkin's lymphoma, head and neck cancer (e.g., head and neck squamous cell carcinoma), primary mediastinal large B-cell lymphoma, urothelial carcinoma, hepatocellular carcinoma, microsatellite instability-high cancer, gastric cancer, esophageal cancer, cervical cancer, Merkel cell carcinoma, kidney cancer, or endometrial cancer. In some embodiments, the second therapeutic agent is MPDL3280A and the cancer being treated is lung cancer (e.g., non-small cell lung cancer or small cell lung cancer), urothelial carcinoma, hepatocellular carcinoma, or breast cancer. In some embodiments, the second therapeutic agent is MEDI4736 and the cancer being treated is lung cancer (e.g., non-small cell lung cancer or small cell lung cancer) or urothelial carcinoma. In some embodiments, the second therapeutic agent is MSB0010718C and the cancer being treated is urothelial carcinoma. In some embodiments, the second therapeutic agent is MSB0010718C and the cancer being treated is melanoma, lung cancer (e.g., non-small cell lung cancer), colorectal cancer, renal cancer, ovarian cancer, pancreatic cancer, gastric cancer, and breast cancer.

[0079] Advantageously, the compounds of the present invention and checkpoint inhibitors can act synergistically to treat cancer.Without wishing to be bound by theory, the synergistic effect between the compounds of the present invention and checkpoint inhibitors can be attributed to the improved efficacy of checkpoint inhibitors associated with BRD9 inhibition-induced downregulation of Foxp3+Treg cells.

[0080] In some embodiments, the anti-cancer therapy is an adoptive T cell transfer (ACT) therapy. In some embodiments, the T cells are activated T cells. The T cells may be modified to express a chimeric antigen receptor (CAR). The CAR-modified T (CAR-T) cells may be generated by any method known in the art. For example, the CAR-T cells may be generated by introducing a suitable expression vector encoding a CAR into the T cells. Prior to the expansion and genetic modification of the T cells, a source of T cells is obtained from the subject. The T cells may be obtained from many sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, umbilical cord blood, thymus tissue, tissue from a site of infection, ascites, pleural effusion, spleen tissue, and tumors. Certain embodiments of the invention may use any number of T cell lines available in the art. In some embodiments, the T cells are autologous T cells. Either before or after genetically modifying the T cells to express a desired protein (e.g., a CAR), the T cells can be engineered to express a desired protein (e.g., a CAR) using methods described, for example, in U.S. Patent Nos. 6,352,694; 6,534,055; 6,905,680; 6,692,964; 5,858,358; 6,887,466; 6,905,681; 7,144,575. Nos. 7,067,318; 7,172,869; 7,232,566; 7,175,843; 5,883,223; 6,905,874; 6,797,514; 6,867,041; and U.S. Patent Application Publication No. 20060121005 can generally be used to activate and expand.

[0081] 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, 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 days, 1-14 days, 1-21 days, or 1-30 days before or after the second therapeutic agent.

[0082] Pharmaceutical Compositions The pharmaceutical compositions described herein are preferably formulated into pharmaceutical compositions for administration to human subjects in a biologically compatible form suitable for administration in vivo.

[0083] The compounds described herein may be used in the form of free base, in the form of salts, in the form of solvates, and as prodrugs. All forms are within the scope of the methods described herein. According to the methods of the present invention, the compounds described or their salts, solvates, or prodrugs may be administered to a patient in various forms depending on the selected route of administration, as will be understood by those skilled in the art. The compounds described herein may be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump, intratumoral, 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.

[0084] The compounds described herein may be administered orally, for example, with an inert diluent or an assimilable edible carrier, or may be enclosed in hard or soft shell gelatin capsules, or may be compressed into tablets, or may be incorporated directly into dietary foods. For oral therapeutic administration, the compounds described herein may be incorporated with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, and wafers. The compounds described herein may also be administered parenterally. Solutions of the compounds described herein may be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions may be prepared in glycerol, liquid polyethylene glycols, DMSO, and mixtures thereof with or without alcohol, and in oils. These preparations may contain a preservative to prevent the growth of microorganisms under ordinary conditions of storage and use. Conventional procedures and ingredients for the selection and preparation of suitable formulations are described, for example, in Remington's Pharmaceutical Sciences (2012, 22nd ed.) and The United States Pharmacopeia: The National Formulary (USP41NF36), published in 2018. Pharmaceutical forms suitable for injection 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 shall be sterile and shall be fluid to the extent that it may be easily administered by syringe. Compositions for nasal administration may be conveniently formulated as aerosols, drops, gels, and powders. Aerosol formulations typically comprise a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent, and are usually presented in single or multiple dose amounts in sterile form in a sealed container, which may take the form of a cartridge or refill for use in a nebulizer device. Alternatively, the sealed container may be a single dispensing device such as a single dose nasal inhaler or an aerosol dispenser with a metering valve intended for disposal after use.If the dosage form comprises an aerosol dispenser, it will contain a propellant, which may be a compressed gas, e.g., compressed air, or an organic propellant, e.g., fluorochlorohydrocarbon. The aerosol dosage form may also take the form of a pump-type atomizer. Suitable compositions for buccal or sublingual administration include tablets, lozenges, and pastilles, in which the active ingredient is formulated with a carrier, e.g., sugar, acacia, tragacanth, gelatin, and glycerin. Compositions for rectal administration are conveniently in the form of a suppository containing a conventional suppository base, such as cocoa butter. The compounds described herein may be administered intratumorally, e.g., as an intratumoral injection. Intratumoral injection is a direct injection into the 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 injection or multiple injections into the tumor, e.g., approximately 1 cm apart. In the case of surgical intervention, the present invention may be used preoperatively, e.g., to remove inoperable tumors. Continuous administration can also be applied where appropriate, for example, by implanting a catheter into the tumor or tumor vasculature.

[0085] The compounds described herein, as described herein, can be administered to animals, e.g., humans, alone or in combination with pharma- ceutically acceptable carriers, the proportions of which will be determined by the solubility and chemical properties of the compound, the chosen route of administration, and standard pharmaceutical practice.

[0086] Dosage The dosage of the compounds described herein and / or compositions comprising the compounds described herein 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 concurrent 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 compounds described herein may be initially administered at a suitable dosage, which may be adjusted as necessary, depending on the clinical response. In general, satisfactory results may be obtained when the compounds described herein are administered to humans at a daily dosage of, for example, 0.05 mg to 3000 mg (measured as solid form). Dose ranges include, for example, 10 to 1000 mg (e.g., 50 to 800 mg). In some embodiments, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mg of the compound are administered.

[0087] Alternatively, the dosage can be calculated using the patient's body weight. For example, the dose of the compound or pharmaceutical composition thereof administered to a patient can be in the range of 0.1 to 100 mg / kg.

[0088] kit The invention also relates to kits that include (a) a pharmaceutical composition comprising an agent that reduces the level and / or activity of BRD9 in a cell or a subject as described herein, and (b) a package insert with instructions for performing any of the methods described herein. In some embodiments, the kit includes (a) a pharmaceutical composition comprising an agent that reduces the level and / or activity of BRD9 in a cell or a subject as described herein, (b) an additional therapeutic agent (e.g., an anti-cancer agent), and (c) a package insert with instructions for performing any of the methods described herein. EXAMPLES

[0089] Example 1 - BRD9 degrading agents deplete BRD9 protein The following examples demonstrate depletion of BRD9 protein in synovial sarcoma cells treated with a BRD9 degrading agent.

[0090] Procedure: Cells were treated with DMSO or the BRD9 degrader, Compound 1 (also known as dBRD9, see Remillard et al, Angew. Chem. Int. Ed. Engl. 56(21):5738-5743 (2017); see structure of Compound 1 below) at the doses and time points indicated.

[0091] [ka]

[0092] Whole-cell extracts were fractionated by SDS-PAGE and transferred to polyvinylidene fluoride membranes using a transfer apparatus according to the manufacturer's protocol (Bio-Rad). After incubation with 5% nonfat milk in TBST (10 mM Tris, pH 8.0, 150 mM NaCl, 0.5% Tween 20) for 60 min, the membranes were incubated overnight at 4 °C with antibodies against BRD9 (1:1,000, Bethyl laboratory A303-781A), GAPDH (1:5,000, Cell Signaling Technology), and / or MBP (1:1,000, BioRad). Membranes were washed three times for 10 min and incubated for at least 1 h with anti-mouse or anti-rabbit antibodies conjugated with either horseradish peroxidase (HRP, Figures 2-3) or IRDye (Figure 4, 1:20,000, LI-COR). Blots were washed three times with TBST and developed with the ECL system according to the manufacturer's protocol (Figures 2-3) or scanned with an Odyssey CLx imaging system (Figure 4).

[0093] Results: Treatment of SYO1 synovial sarcoma cells with the BRD9 degrader compound 1 resulted in dose-dependent (Figure 1) and time-dependent (Figure 2) depletion of BRD9 in the cells. Furthermore, as shown in Figure 3, depletion of BRD9 by compound 1 was also reproduced in a non-synovial sarcoma cell line (293T) and could persist for at least 5 days.

[0094] Example 2 - Inhibition of proliferation of synovial cell lines by BRD9 inhibitors and BRD9 degraders The following examples demonstrate that BRD9 degraders and inhibitors selectively inhibit the proliferation of synovial sarcoma cells.

[0095] procedure: Cells were treated with DMSO or the BRD9 degrader, compound 1, at the indicated concentrations, and proliferation was monitored by measuring culture confluency over time using an IncuCyte live cell analysis system from day 7 to day 14 (Figure 4). Growth medium and compounds were refreshed every 3-4 days.

[0096] Cells were seeded in 12-well plates and treated with DMSO, 1 μM BRD9 inhibitor, compound 2 (also known as BI-7273, see Martin et al, J Med Chem. 59(10):4462-4475 (2016); see structure of compound 2 below), or 1 μM BRD9 degrader, compound 1.

[0097] [ka]

[0098] Cell numbers were optimized for each cell line. Growth medium and compounds were refreshed every 3–5 days. SYO1, Yamato, A549, 293T, and HS-SY-II cells were fixed and stained on day 11. ASKA cells were fixed and stained on day 23. Staining was performed by incubating with crystal violet solution (0.5 g crystal violet, 27 ml 37% formaldehyde, 100 mL 10x PBS, 10 mL methanol, 863 dH20 to 1 L) for 30 min, followed by washing three times with water and drying the plates at room temperature for at least 24 h. Plates were then scanned with an Odyssey CLx imaging system (Figure 5).

[0099] Cells were seeded in 96-well ultra-low cluster plates (Costar, #7007) in 200 μL complete medium and treated with DMSO, staurosporine, or the BRD9 degrader, compound 1, at the indicated doses on day 2 ( FIG. 2C ). Medium and compounds were changed every 5 days, and cell colonies were imaged on day 14.

[0100] Results: As shown in Figures 4, 5, and 6, treatment of synovial sarcoma cell lines (SYO1, Yamato, HS-SY-II, and ASKA) with a BRD9 inhibitor, compound 2, or a BRD9 degrader, compound 1, resulted in inhibition of cell proliferation, but not of non-synovial control cancer cell lines (293T, A549, G401).

[0101] Example 3 - Selective inhibition of proliferation of synovial cell lines by BRD9 degraders and BRD9 binders The following examples demonstrate that BRD9 degraders and binding agents selectively inhibit the proliferation of synovial sarcoma cells.

[0102] Procedure: Cells were seeded in 6-well or 12-well plates and treated daily with a BRD9 degrader (compound 1), a bromo-domain BRD9 binder (compound 2), an E3 ligase binder (lenalidomide), DMSO, or staurosporine (positive control for cell killing) at the indicated concentrations. Cell numbers were optimized for each cell line. Growth medium was refreshed every 5 days. By day 14, medium was removed, cells were washed with PBS, and stained with 500 μL of 0.005% (w / v) crystal violet in 25% (v / v) methanol for at least 1 h at room temperature. Plates were then scanned with an Odyssey CLx imaging system.

[0103] Results: As shown in Figures 7 and 8, treatment of synovial sarcoma cell lines (SYO1, HS-SY-II, and ASKA) with Compound 1 or Compound 2 resulted in inhibition of cell proliferation, but not of non-synovial control cancer cell lines (RD, HCT116, and Calu6). Overall, Compound 1 showed the most significant proliferation inhibition in all synovial cell lines.

[0104] Example 4 - Inhibition of cell proliferation in synovial sarcoma cells The following examples demonstrate that BRD9 degraders inhibit cell proliferation and induce apoptosis in synovial sarcoma cells.

[0105] Procedure: SYO1 cells were treated with DMSO, a BRD9 degrader (compound 1) at 200 nM or 1 μM, or an E3 ligase binder (lenalidomide) at 200 nM for 8 or 13 days. Compounds were refreshed every 5 days. Cell cycle analysis was performed using the Click-iT™ Plus EdU flow cytometry assay (Invitrogen). Apoptosis assays were performed using the Annexin V-FITC Apoptosis Detection Kit (Sigma A9210). The assays were performed according to the manufacturer's protocol.

[0106] Results: As shown in Figures 9-12, treatment with Compound 1 for 8 or 13 days resulted in a decrease in the number of cells in the S phase of the cell cycle compared to DMSO and lenalidomide. Treatment with Compound 1 for 8 days also resulted in an increase in the number of early and late apoptotic cells compared to DMSO controls.

[0107] Example 5 - Composition for SS18-SSX1-BAF The following examples demonstrate the identification of BRD9 as a component of the SS18-SSX-containing BAF complex.

[0108] Procedure: A stable 293T cell line expressing HA-SS18SSX1 was generated using lentiviral integration. SS18-SSX1-containing BAF complexes were subjected to affinity purification and subsequent mass spectrometry analysis to reveal SS18-SSX1 interacting proteins.

[0109] Results: As shown in Figure 13, the BAF complex containing the SS18-SSX fusion protein also contained BRD9. More than five unique peptides were identified for ARID1A (95 peptides), ARID1B (77 peptides), SMARCC1 (69 peptides), SMARCD1 (41 peptides), SMARCD2 (37 peptides), DPF2 (32 peptides), SMARCD3 (26 peptides), ACTL6A (25 peptides), BRD9 (22 peptides), DPF1 isoform 2 (18 peptides), DPF3 (13 peptides), and ACTL6B (6 peptides).

[0110] Example 6 - Preparation of 4-[6-(azetidin-1-yl)-2-methyl-1-oxo-2,7-naphthyridin-4-yl]-2,6-dimethoxybenzaldehyde (Intermediate H)

[0111] [ka]

[0112] Step 1: Preparation of 6-chloro-4-methylpyridine-3-carboxamide (Intermediate B)

[0113] [ka]

[0114] To a stirred mixture of 6-chloro-4-methylpyridine-3-carboxylic acid (20.00 g, 116.564 mmol, 1.00 equiv) and NH4Cl (62.35 g, 1.17 mol, 10.00 equiv) in dichloromethane (DCM; 400 mL) was added DIEA (22.60 g, 174.846 mmol, 3.00 equiv). After stirring for 5 min, HATU (66.48 g, 174.846 mmol, 1.50 equiv) was added in portions. The resulting mixture was stirred for 3 h at room temperature. The resulting mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with 1 / 1 to 3 / 2 petroleum ether / ethyl acetate (PE / EtOAc) to give 6-chloro-4-methylpyridine-3-carboxamide (18.30 g, 61.3%) as a yellow solid. LCMS(ESI)m / z:[M+H] + =171.

[0115] Step 2: Preparation of 6-chloro-N-[(1E)-(dimethylamino)methylidene]-4-methylpyridine-3-carboxamide (Intermediate C)

[0116] [ka]

[0117] To a stirred mixture of 6-chloro-4-methylpyridine-3-carboxamide (18.30 g, 107.268 mmol, 1.00 equiv) in 2-methyltetrahydrofuran (100 mL) was added DMF-DMA (19.17 g, 160.903 mmol, 1.50 equiv) at 80° C. under nitrogen atmosphere and stirred for an additional 1 h. The mixture was then cooled and concentrated to give 6-chloro-N-[(1E)-(dimethylamino)methylidene]-4-methylpyridine-3-carboxamide (26.3 g, 91.3%) as a crude yellow solid, which was used directly without further purification. LCMS (ESI) m / z: [M+H] + =226.

[0118] Step 3: Preparation of 6-chloro-2H-2,7-naphthyridin-1-one (Intermediate D)

[0119] [ka]

[0120] To a stirred mixture of 6-chloro-N-[(1E)-(dimethylamino)methylidene]-4-methylpyridine-3-carboxamide (26.30 g) in THF (170.00 mL) was added t-BuOK (174.00 mL, 1 mol / L in THF). The resulting solution was stirred at 60° C. under nitrogen atmosphere for 30 min. The mixture was then cooled and concentrated under reduced pressure. The crude solid was washed with saturated NaHCO3 solution (100 mL) and collected to give 6-chloro-2H-2,7-naphthyridin-1-one (14.1 g, 67.0%) as a pink solid, which was used directly without further purification. LCMS (ESI) m / z: [M+H] + =181.

[0121] Step 4: Preparation of 6-chloro-2-methyl-2,7-naphthyridin-1-one (Intermediate E)

[0122] [ka]

[0123] To a stirred mixture of 6-chloro-2H-2,7-naphthyridin-1-one (14.10 g, 78.077 mmol, 1.00 equiv) in anhydrous THF (280.00 mL) was added NaH (9.37 g, 234.232 mmol, 3.00 equiv, 60%) in portions at 0° C. After 10 min, MeI (33.25 g, 234.232 mmol, 3.00 equiv) was added at 0° C. and the mixture was allowed to stir at 0° C. for 10 min, then the mixture was allowed to stir at room temperature for 12 h. The resulting mixture was concentrated under reduced pressure. The crude solid was slurried with water (100 mL) and the solid was filtered and collected to give 6-chloro-2-methyl-2,7-naphthyridin-1-one (14.6 g, 94.1%) as a yellow solid, which was used directly without further purification. LCMS(ESI)m / z:[M+H] + =195.

[0124] Step 5: Preparation of 4-bromo-6-chloro-2-methyl-2,7-naphthyridin-1-one (Intermediate F)

[0125] [ka]

[0126] To a stirred mixture of 6-chloro-2-methyl-2,7-naphthyridin-1-one (8.00 g, 41.106 mmol, 1.00 equiv) in DMF (160.00 mL) was added NBS (8.78 g, 49.327 mmol, 1.20 equiv) and the resulting mixture was stirred at 90° C. for 2 h. The reaction mixture was cooled, diluted with DCM (150 mL) and washed with water (3×100 mL). The organic layer was dried and concentrated. The residue was then slurried with EtOAc (20 mL) and the slurry was filtered. The filter cake was washed with EtOAc (20 mL) to give 4-bromo-6-chloro-2-methyl-2,7-naphthyridin-1-one (6.32 g, 55.7%) as a white solid, which was used directly without further purification. LCMS (ESI) m / z: [M+H] + =273.

[0127] Step 6: Preparation of 6-(azetidin-1-yl)-4-bromo-2-methyl-2,7-naphthyridin-1-one (Intermediate G)

[0128] [ka]

[0129] To a solution of 4-bromo-6-chloro-2-methyl-2,7-naphthyridin-1-one (5.00 g, 18.281 mmol, 1.00 equiv.) and azetidine hydrochloride (3.2 g, 54.843 mmol, 3 equiv.) in DMSO (50.00 mL) was added K2CO3 (12.6 g, 91.404 mmol, 5 equiv.). The resulting solution was stirred at 130° C. for 2 h. The resulting mixture was cooled, diluted with water (100 mL), and then extracted with EtOAc (3×100 mL). The combined organic layers were washed with saturated NaCl solution (3×50 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to give 6-(azetidin-1-yl)-4-bromo-2-methyl-2,7-naphthyridin-1-one (3.7 g, 68.8%) as a grey solid. This was used directly without further purification. LCMS (ESI) m / z: [M+H] + =294.

[0130] Step 7: Preparation of 4-[6-(azetidin-1-yl)-2-methyl-1-oxo-2,7-naphthyridin-4-yl]-2,6-dimethoxybenzaldehyde (Intermediate H)

[0131] [ka]

[0132] To a solution of 6-(azetidin-1-yl)-4-bromo-2-methyl-2,7-naphthyridin-1-one (1.42 g, 4.827 mmol, 1.00 equiv) and 4-formyl-3,5-dimethoxyphenylboronic acid (1.52 g, 7.241 mmol, 1.5 equiv) in dioxane (16.00 mL) and H2O (4.00 mL) was added Pd(dppf)Cl2 (353.2 mg, 0.483 mmol, 0.1 equiv) and Cs2CO3 (3.15 g, 9.655 mmol, 2 equiv) and the resulting solution was stirred at 70 °C for 2 h. The resulting mixture was cooled and concentrated under reduced pressure. The residue was slurried with water (30 mL), filtered, and the filter cake was collected. The solid was further slurried with MeOH (30 mL) and filtered. The solid was collected to give 4-[6-(azetidin-1-yl)-2-methyl-1-oxo-2,7-naphthyridin-4-yl]-2,6-dimethoxybenzaldehyde (1.42 g, 77.5%) as a grey solid. LCMS (ESI) m / z: [M+H] + =380.

[0133] Example 7 - Preparation of 3-[1-oxo-6-[7-(piperidin-4-ylmethyl)-2,7-diazaspiro[3.5]nonan-2-yl]-3H-isoindol-2-yl]piperidine-2,6-dione (Intermediate P)

[0134] [ka]

[0135] Step 1: Preparation of methyl 5-bromo-2-(bromomethyl)benzoate (Intermediate J).

[0136] [ka]

[0137] To a stirred mixture of methyl 5-bromo-2-methylbenzoate (50.00 g, 218.271 mmol, 1.00 equiv) in CCl4 (500.00 mL) was added NBS (38.85 g, 218.271 mmol, 1.00 equiv) and BPO (5.59 g, 21.827 mmol, 0.10 equiv). After stirring overnight at 80° C., the mixture was purified by silica gel column chromatography eluting with PE / EtOAc (50:1) to give methyl 5-bromo-2-(bromomethyl)benzoate (67 g, 74.75%) as a yellow oil. LCMS (ESI) m / z: [M+H] + =307.

[0138] Step 2: Preparation of tert-butyl 4-(6-bromo-1-oxo-3H-isoindol-2-yl)-4-carbamoylbutanoate (Intermediate K)

[0139] [ka]

[0140] To a stirred mixture of methyl 5-bromo-2-(bromomethyl)benzoate (67.00 g, 217.554 mmol, 1.00 equiv) and tert-butyl (4S)-4-amino-4-carbamoylbutanoate hydrochloride (62.32 g, 261.070 mmol, 1.20 equiv) in DMF (100.00 mL) was added DIEA (112.47 g, 870.217 mmol, 4 equiv). After stirring overnight at 80° C., the mixture was concentrated under reduced pressure. Water (200 mL) was added to the residue and stirred for 1 h at room temperature. The resulting mixture was filtered and water (100 mL) was added to the filter cake and stirred. The precipitated solid was collected by filtration and washed with water (3×30 mL). This gave tert-butyl 4-(6-bromo-1-oxo-3H-isoindol-2-yl)-4-carbamoylbutanoate (55 g, 60.46%) as an off-white solid. LCMS (ESI) m / z: [M+H] + =397.

[0141] Step 3: Preparation of tert-butyl 2-[2-[4-(tert-butoxy)-1-carbamoyl-4-oxobutyl]-3-oxo-1H-isoindol-5-yl]-2,7-diazaspiro[3.5]nonane-7-carboxylate (Intermediate L)

[0142] [ka]

[0143] To a stirred solution of tert-butyl 4-(6-bromo-1-oxo-3H-isoindol-2-yl)-4-carbamoylbutanoate (10.00 g, 25.172 mmol, 1.00 equiv.) and tert-butyl 2,7-diazaspiro[3.5]nonane-7-carboxylate hydrochloride (8.60 g, 32.723 mmol, 1.30 equiv.) in dioxane (200.00 mL) was added Cs2CO3 (24.60 g, 75.516 mmol, 3.00 equiv.) and RuPhos palladacycle 3rd generation (2.11 g, 2.517 mmol, 0.10 equiv.). After stirring overnight at 100 °C under nitrogen atmosphere, the resulting mixture was filtered while hot and the filter cake was washed with 1,4-dioxane (3 x 50 mL). The filtrate was concentrated under reduced pressure to give tert-butyl 2-[2-[4-(tert-butoxy)-1-carbamoyl-4-oxobutyl]-3-oxo-1H-isoindol-5-yl]-2,7-diazaspiro[3.5]nonane-7-carboxylate (21 g, crude) as a black solid. LCMS (ESI) m / z: [M+H] + =543.

[0144] Step 4: Preparation of tert-butyl 2-[2-(2,6-dioxopiperidin-3-yl)-3-oxo-1H-isoindol-5-yl]-2,7-diazaspiro[3.5]nonane-7-carboxylate (Intermediate M)

[0145] [ka]

[0146] To a stirred mixture of tert-butyl 2-[2-[(1S)-4-(tert-butoxy)-1-carbamoyl-4-oxobutyl]-3-oxo-1H-isoindol-5-yl]-2,7-diazaspiro[3.5]nonane-7-carboxylate (13.68 g, 25.208 mmol, 1.00 equiv) in THF (100.00 mL) was added t-BuOK in THF (25.00 mL, 25.000 mmol, 0.99 equiv). The resulting mixture was stirred for 2 h at room temperature. The mixture was acidified with 1M HCl (aq) to pH 6 and then neutralized with saturated NaHCO3 (aq) to pH 7. The resulting mixture was extracted with EtOAc (3 x 200 mL). The combined organic layers were concentrated under reduced pressure. This gave tert-butyl 2-[2-(2,6-dioxopiperidin-3-yl)-3-oxo-1H-isoindol-5-yl]-2,7-diazaspiro[3.5]nonane-7-carboxylate (7.8 g, 59.43%) as a yellow solid. LCMS (ESI) m / z: [M+H] + =469.

[0147] Step 5: Preparation of 3-(6-[2,7-diazaspiro[3.5]nonan-2-yl]-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione (Intermediate N)

[0148] [ka]

[0149] To a stirred mixture of tert-butyl 2-[2-(2,6-dioxopiperidin-3-yl)-3-oxo-1H-isoindol-5-yl]-2,7-diazaspiro[3.5]nonane-7-carboxylate (7.80 g, 16.647 mmol, 1.00 equiv) in DCM (10.00 mL) was added trifluoroacetic acid (TFA; 5.00 mL). After stirring for 2 h at room temperature, the resulting mixture was concentrated under vacuum. This afforded 3-(6-[2,7-diazaspiro[3.5]nonan-2-yl]-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione (6 g, 92.93%) as a pale yellow solid. LCMS (ESI) m / z: [M+H] + =369.

[0150] Step 6: Preparation of tert-butyl 4-([2-[2-(2,6-dioxopiperidin-3-yl)-3-oxo-1H-isoindol-5-yl]-2,7-diazaspiro[3.5]nonan-7-yl]methyl)piperidine-1-carboxylate (Intermediate O)

[0151] [ka]

[0152] To a stirred solution of 3-(6-[2,7-diazaspiro[3.5]nonan-2-yl]-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione (4.00 g, 8.685 mmol, 1.00 equiv, 80%) and tert-butyl 4-formylpiperidine-1-carboxylate (1.48 g, 6.939 mmol, 0.80 equiv) in DMF (20.00 mL) was added NaBH(OAc)3 (3.68 g, 17.363 mmol, 2.00 equiv) at room temperature. The resulting mixture was stirred for 2 h at room temperature. The reaction was quenched with water at room temperature. The resulting mixture was purified by reverse flash chromatography using the following conditions: column, C18 silica gel; mobile phase, CH3CN in water (0.1% FA), 0-100% gradient in 40 min; detector, UV 254 nm. This gave tert-butyl 4-([2-[2-(2,6-dioxopiperidin-3-yl)-3-oxo-1H-isoindol-5-yl]-2,7-diazaspiro[3.5]nonan-7-yl]methyl)piperidine-1-carboxylate (2.8 g, 51.29%) as a dark yellow solid. LCMS (ESI) m / z: [M+H] + =566.

[0153] Step 7: Preparation of 3-[1-oxo-6-[7-(piperidin-4-ylmethyl)-2,7-diazaspiro[3.5]nonan-2-yl]-3H-isoindol-2-yl]piperidine-2,6-dione (Intermediate P)

[0154] [ka]

[0155] To a stirred mixture of tert-butyl 4-([2-[2-(2,6-dioxopiperidin-3-yl)-3-oxo-1H-isoindol-5-yl]-2,7-diazaspiro[3.5]nonan-7-yl]methyl)piperidine-1-carboxylate (2.80 g, 4.949 mmol, 1.00 equiv) in DCM (5.00 mL) was added TFA (2.00 mL). The mixture was stirred for 2 h at room temperature. The resulting mixture was concentrated under reduced pressure to give 3-[1-oxo-6-[7-(piperidin-4-ylmethyl)-2,7-diazaspiro[3.5]nonan-2-yl]-3H-isoindol-2-yl]piperidine-2,6-dione (3.9 g, crude) as a yellow solid. LCMS (ESI) m / z: [M+H] + =466.

[0156] Example 8 - Preparation of 3-[6-(7-[[1-([4-[6-(azetidin-1-yl)-2-methyl-1-oxo-2,7-naphthyridin-4-yl]-2,6-dimethoxyphenyl]methyl)piperidin-4-yl]methyl]-2,7-diazaspiro[3.5]nonan-2-yl)-1-oxo-3H-isoindol-2-yl]piperidine-2,6-dione TFA salt (Compound D1 TFA salt)

[0157] [ka]

[0158] A solution of 3-[1-oxo-6-[7-(piperidin-4-ylmethyl)-2,7-diazaspiro[3.5]nonan-2-yl]-3H-isoindol-2-yl]piperidine-2,6-dione (4.5 g, 10.52 mmol, 1.00 equiv.) and 4-[6-(azetidin-1-yl)-2-methyl-1-oxo-2,7-naphthyridin-4-yl]-2,6-dimethoxybenzaldehyde (4.0 g, 10.52 mmol, 1.00 equiv.) and titanium(IV) isopropoxide (3.0 g, 10.52 mmol, 1.00 equiv.) in DMSO (100 mL) was stirred at room temperature for 3 h. NaBH(OAc)3 (8.92 g, 42.08 mmol, 4.00 equiv) was then added to the resulting solution in several batches, and the resulting mixture was stirred at room temperature overnight. The reaction was quenched by the addition of water (30 mL), and then the solution was filtered. The filter cake was washed with water and acetonitrile. The filtrate was then concentrated in vacuum. The crude product was purified by reverse phase flash chromatography with the following conditions (Column: AQ C18 column, 50×250 mm 10 μm; Mobile phase A: water (TFA 0.1%), Mobile phase B: ACN; Flow rate: 100 mL / min; Gradient: 5B to 25B in 35 min; 254 / 220 nm). Pure fractions were evaporated to dryness to give 3-[6-(7-[[1-([4-[6-(azetidin-1-yl)-2-methyl-1-oxo-2,7-naphthyridin-4-yl]-2,6-dimethoxyphenyl]methyl)piperidin-4-yl]methyl]-2,7-diazaspiro[3.5]nonan-2-yl)-1-oxo-3H-isoindol-2-yl]piperidine-2,6-dione TFA salt (3.2 g, 32.3%) as a white solid. 1H NMR(400MHz,DMSO-d6)δ10.96(s,1H),9.01(s,1H),7.59(s,1H),7.36(d,J=8.0Hz,1H),6.72(s,2H), 6.68(d,J=8.1Hz,2H),6.20(s,1H),5.07(dd,J=13.3,5.1Hz,1H),4.35-4.13(m,2H),4.06-3.95(m,4 H),3.80(s,6H),3.57(s,4H),3.47(s,5H),2.97-2.75(m,3H),2.70-2.55(m,1H),2.44-2.16(m,7H), 2.13-1.88(m,5H),1.80-1.67(m,4H),1.61(d,J=12.4Hz,2H),1.53-1.33(m,1H),1.13-0.94(m,2H). LCMS(ESI)m / z:[M+H] + =829.55.

[0159] The enantiomers of compound D1 were separated by supercritical fluid chromatography on a chiral support to produce compound S-D1 and compound R-D1. Example 9 - Preparation of Compound D2 Compound D2 was prepared using the appropriate starting materials in a similar manner to the procedures described in the above examples.

[0160] Compound D2: 1H NMR(300MHz,DMSO-d6)δ10.98(s,1H),9.02(d,J=0.7Hz,1H),7.63(d,J=2.3Hz,1H),7.41(d,J=8.8Hz,1H),6.88(s, 2H),6.70(h,J=2.4Hz,2H),6.24(d,J=6.0Hz,1H),5.08(dd,J=13.2,5.1Hz,1H),4.41-4.14(m,4H),4.04(t,J=7.4Hz ,4H),3.91(s,1H),3.70(d,J=22.5Hz,4H),3.50(s,3H),3.45(s,1H),3.22(s,1H),3.14-2.82(m,6H),2.60(d,J=16 .2Hz,1H),2.55(s,2H),2.44-2.28(m,3H),2.18-2.05(m,3H),1.97(t,J=13.9Hz,5H),1.51(q,J=12.2,11.1Hz,2H). LCMS(ESI)m / z:[M+H] + =835.45.

[0161] Example 10 - SYO1 BRD9 NanoLuc Degradation Assay This example demonstrates the ability of compounds of the present disclosure to degrade nanoluciferase-BRD9 fusion protein in a cell-based degradation assay.

[0162] Procedure: A stable SYO-1 cell line expressing 3xFLAG-NLuc-BRD9 was generated. On day 0, cells were seeded into each well of a 384-well cell culture plate in 30 μL medium. The seeding density was 8000 cells / well. On day 1, cells were treated with 30 nL DMSO or 30 nL of 3-fold serial DMSO dilutions of compounds (10 points with 1 μM final highest dose in duplicate). Plates were then incubated for 6 hours in a standard tissue culture incubator and equilibrated at room temperature for 15 minutes. Nanoluciferase activity was measured by adding 15 μL of freshly prepared Nano-Glo Luciferase Assay Reagent (Promega N1130), shaking the plate for 10 minutes, and reading bioluminescence using an EnVision reader.

[0163] Results: The inhibition rate (%) was calculated using the following formula: % inhibition = 100 × (Lum HC -Lum サンプル ) / (Lum HC -Lum LC DMSO-treated cells are taken as the high control (HC) and 1 μM of a known BRD9 degrader standard treated cells are taken as the low control (LC). The data were fitted to a four parameter, non-linear curve fit to obtain IC values ​​as shown in Table 2. 50 The (μM) values ​​were calculated. As shown by the results in Table 2, some compounds of the present disclosure had IC 50 The present invention provides methods for treating BRD9-associated disorders, including the treatment of cancer, in which BRD9 is a cytotoxic and / or anti-inflammatory agent, and / or a therapeutic agent for treating cancer, in accordance with the present invention.

[0164] [Table 2]

[0165] Example 11 - Degradation of BRD9 inhibits synovial sarcoma tumor growth in vivo. Methods: NOD SCID mice (Beijing Anikeeper Biotech, Beijing) were inoculated subcutaneously into the right flank with a single-cell suspension of SYO-1 human biphasic synovial sarcoma tumor cells (5 × 106) in 100 μL of Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS). The average tumor size was approximately 117 mm 3 Upon reaching 10 mg / kg, mice were randomized to either the control group [10% dimethylsulfoxide (DMSO), 40% polyethylene glycol (PEG400) and 50% water] or treatment group D1. Mice were dosed daily via the intraperitoneal (ip) route for 3 weeks. All dose volumes were adjusted by body weight in terms of mg / kg.

[0166] Results: Treatment with compound D1 at 1 mg / kg resulted in tumor growth inhibition, as shown in Figure 14. All treatments were well tolerated based on the % body weight change observed.

[0167] Example 12 - Compound D1 causes degradation of BRD9 in synovial sarcoma tumors in vivo. Methods: Mice were treated with D1, 1 mg / kg, ip for 4 weeks. Mice were then euthanized and tumors were harvested at 8, 72, and 168 hours after the last dose. Tumors were lysed with 1x RIPA lysis buffer (Boston BioProducts, BP-115D) containing protease and phosphatase protein inhibitors (Roche Applied Science No. 04906837001&05892791001). Equal amounts of lysates (30 μg) were loaded onto 4-12% Bis-Tris Midi protein gels in 1x MOPS buffer; samples were run at 120 V for 120 min. Proteins were transferred to membranes using TransBlot at 250 mA for 150 min, after which the membranes were blocked with Odyssey blocking buffer for 1 h at room temperature. Membranes were hybridized with primary antibodies overnight in a refrigerator. Images were acquired using a Li-COR imaging system (Li-COR Biotechnology, Lincoln, Nebraska).

[0168] Table 3 shows the detection antibody information.

[0169] [Table 3]

[0170] Results: As shown in FIG. 15, treatment with compound D1 at 1 mg / kg resulted in complete degradation of the BRD9 target up to 168 hours post-dose. Example 13 - Effect of compounds S-D1 and R-D1 on synovial sarcoma cells Methods. Synoial sarcoma cells were plated at 500-100k cells / well in 6-well plates and treated with graded concentrations of BRD9 degraders (highest concentration 10 nM, diluted 1:3) at 37°C for two time points the following day. Cells were then harvested, washed with cold PBS, and frozen into cell pellets at -80°C. Lysates were prepared by resuspending thawed pellets in 1x RIPA Lysis and Extraction Buffer (Thermo Fisher, Catalog No. 89900) containing 1x Halt™ Protease and Phosphatase Inhibitor Cocktail, EDTA-free (Thermo Fisher, Catalog No. 78441) and 1:1000 diluted Pierce™ Universal Nuclease for Cell Lysis 25ku (Thermo Fisher, Catalog No. 88700). Lysates were incubated on ice for 10 min and then centrifuged at maximum speed (15,000 rpm) for 10 min at 4°C. Samples were then analyzed for total protein using the BCA Protein Quantitation Assay and diluted to 1 μg / μL with lysis buffer and 1× NuPAGE™ LDS Sample Buffer (4×) (Thermo Fisher, Cat. No. NP0007) and 1× DTT from a 30× stock (Cell Signaling Technologies, Cat. No. 14265S). Samples with 20-25ug total protein were loaded onto a 4-12% Bis-Tris Mini gel with 1× MES running buffer and run at 150V for 45 minutes. Gels were transferred onto nitrocellulose blots using the Trans-Blot® Turbo™ transfer system (semi-dry) at 25V for 10 minutes (high MW setting). Blots were blocked with 5% milk in TBST for 1 hour and probed with BRD9 antibody (for SYO1, Bethyl Labs, Cat. No. A303-781A, 1:750; for ASKA, Cell Signaling Technologies, Cat. No. 71232S) and beta-actin antibody (Cell Signaling Technologies, Cat. No. 3700, 1:2000) overnight at 4°C.The next day, the blots were washed three times with TBST and probed with 1:5000 IRDye® 680LT goat anti-rabbit IgG secondary antibody (LiCOR, Cat. No. 926-68021) and 1:10000 IRDye® 800CW goat anti-mouse IgG secondary antibody (LiCOR, Cat. No. 926-32210) in LiCOR Odyssey® Blocking Buffer (TBS) for 1 hour at room temperature. The blots were washed three times with TBST and scanned using a LiCOR Odyssey® CLx Imaging System at wavelengths of 700 nM and 800 nM. Western blot signals were quantified using the same analysis program included in the same machine. BRD9 signal was quantified by normalizing to beta-actin signal, and all samples were normalized to DMSO set as 100% signal.

[0171] To evaluate the interconversion between Enantiomer 1 and Enantiomer 2 in cell culture media, the following study was performed. Enantiomer 1 and Enantiomer 2 (each 40 μM in DMSO) were spiked into cell culture media (DMEM+Glutamax+10% FBS) at a final concentration of 0.2 μM and incubated in duplicate at 37° C. and 5% CO2. At the indicated time points, aliquots (50 μL) were removed and processed by adding 150 μL of acetonitrile containing 0.1% formic acid and an internal standard for LC / MS-MS analysis. Both Enantiomer 1 and Enantiomer 2 peak areas were determined for each sample using chiral specific analytical methods. The results are summarized in Table 5 below.

[0172] Results. To evaluate the BRD9 degradation activity of the two enantiomers, degradant treatment and subsequent Western blot experiments were performed using two synovial sarcoma cell lines (SYO-1 and ASKA). Significantly more potent BRD9 degradation activity was observed with enantiomer 2, which was significantly higher than that observed with fit DC in SYO-1 at a 1-hour treatment time. 50The DC value for Enantiomer 2 at 30 min was 0.092 nM compared to 2.8 nM for Enantiomer 1 (Figures 16, 17, and 18 and Table 4). An even more dramatic difference in ASKA cells was the DC 50 A DC of 0.34 nM is evident, while there is no discernible activity of Enantiomer 1 up to 10 μM at the same time point (Figures 20, 21, and 22 and Table 4). The difference decreases to about 32-fold in ASKA at the 2-hour time point, with fitted DC DC values ​​for Enantiomer 1 and Enantiomer 2. 50 The values ​​are 0.38 nM and 0.012 nM, respectively (Table 4). The difference further decreases to about 3-fold by 6 hours in SYO1. Enantiomer 2 performs slightly better than its racemic parent compound D1 in degrading BRD9, but is overall comparable under the same test conditions (Figures 16 and 17). BRD9 degradation activity becomes very similar at the 24 hour time point for all three compounds (Figure 19). In summary, enantiomer 2 is much more potent at degrading endogenous BRD9 protein in two synovial sarcoma cell lines at early time points, whereas enantiomer 1 is nearly inactive or has a greatly reduced degradative potency. However, the difference in potency decreases over time and is almost gone by 24 hours.

[0173] [Table 4]

[0174] Epimerization of chiral centers in thalidomide or other IMiD drugs and their derivatives has been reported. The acidic hydrogens of the chiral centers can be scrambled under physiological or neutral pH conditions. To investigate the chiral stability under cellular assay conditions for these degraders, we performed a time course study on enantiomer 1 and enantiomer 2 in cell culture medium at 37°C. There is no detectable enantiomer 2 in the enantiomer 1 samples at time 0 or 0.5 h. However, substantial enantiomer 2 was detected at later time points, accounting for 12% and 30% of the total at 2 and 6 h, respectively (Table 5). Similarly, enantiomer 2 was converted to enantiomer 1 over time, with its effective concentration decreasing to 63% at 6 h (Table 5). These data indicate that the epimerization rate is relatively fast under cellular assay conditions, suggesting that the time-dependent BRD9 degradation activity of enantiomer 1 is likely due to the converted enantiomer 2. Overall, these data indicate that enantiomer 2 is the active enantiomer in degrading BRD9 in cells.

[0175] [Table 5]

[0176] Example 14 - Effect of Compounds S-D1 and R-D1 on Synovial Sacroma Cells Methods. SYO-1 tumor cells were maintained in vitro as adherent cells in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum at 37°C in an atmosphere of 5% CO2 in air. Cells growing in the exponential phase were harvested and counted for tumor inoculation. BALB / c nude mice (Shanghai Lingchang biological science) were inoculated with 5×10 6 Treatment- (listed in the table below) resulted in a mean tumor size of 499 mm 3 The treatment was started on day 19 after tumor inoculation, when tumor

[0177] [Table 6]

[0178] Mice were treated with racemic D1, 1 mg / kg, ip for 4 weeks, mice were euthanized, and tumors were collected 1, 4, 8, 24, 48, and 72 hours after the last dose. Tumors were lysed with 1× RIPA lysis buffer (Boston BioProducts, BP-115D) containing protease and phosphatase protein inhibitors (Roche Applied Science No. 04906837001&05892791001). Equal amounts of lysates (30 μg) were loaded onto 4-12% Bis-Tris Midi protein gels in 1× MOPS buffer; samples were run at 120 V for 120 min. Proteins were transferred to membranes (NC) using TransBlot at 250 mA for 150 min, after which the membranes were blocked with Odyssey blocking buffer for 1 h at room temperature. Membranes were hybridized with primary antibodies overnight in a refrigerator. Images were acquired using a Li-COR imaging system (Li-COR Biotechnology, Lincoln, Nebraska).

[0179] [Table 7]

[0180] Results. The pharmacodynamic activity of Enantiomer 1, Enantiomer 2, and racemate D1 were evaluated in the SYO-1 xenograft model. Enantiomer 2 demonstrated significant activity as assessed by BRD9 protein levels using a Western blot assay. Figure 23. Enantiomer 2 degraded BRD9 up to 72 hours after a single dose. Enantiomer 1 was inactive and did not degrade BRD9 protein. These results suggest that Enantiomer 2 is as potent as racemate D1.

[0181] Other embodiments 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 any term in this application is found to be defined differently in a document incorporated herein by reference, the definition provided herein controls for that term.

[0182] While the invention has been described in connection with specific embodiments thereof, it will be understood that further modifications are possible, and this application is intended to cover any variations, uses, or adaptations of the invention generally in accordance with the principles of the invention and including departures from the present disclosure that may apply to the essential features described above within the scope of known or customary practice in the art to which the invention pertains, and are within the scope of the appended claims.

[0183] Other embodiments are found in the claims. The technical ideas that can be understood from the above-described embodiment will be described below as supplementary notes. [Appendix 1] A compound having the structure: [ka] or a pharma- ceutically acceptable salt thereof. [Appendix 2] The compound has the following structure: [ka] or a pharma- ceutically acceptable salt thereof. [Appendix 3] The compound has the following structure: [ka] or a pharma- ceutically acceptable salt thereof. [Appendix 4] The compound has the following structure: [ka] or a pharma- ceutically acceptable salt thereof. [Appendix 5] A compound having the structure: [ka] or a pharma- ceutically acceptable salt thereof and a pharma- ceutically acceptable excipient. [Appendix 6] The pharmaceutical composition comprises a compound having the structure: [ka] or a pharma- ceutical composition comprising the compound of formula (I) or a pharma- ceutical salt thereof and a pharma- ceutical acceptable excipient. [Appendix 7] The pharmaceutical composition comprises a compound having the structure: [ka] or a pharma- ceutical composition according to claim 6, comprising a pharma- ceutical acceptable salt thereof and a pharma- ceutical acceptable excipient. [Appendix 8] The pharmaceutical composition comprises a compound having the structure: [ka] or a pharma- ceutical composition according to claim 6, comprising a pharma- ceutical acceptable salt thereof and a pharma- ceutical acceptable excipient. [Appendix 9] A method for inhibiting the level of BRD9 in a cell, comprising contacting the cell with an effective amount of a compound described in any one of Appendix 1 to 4, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described in any one of Appendix 5 to 8. [Appendix 10] A method for inhibiting the activity of BRD9 in a cell, the method comprising contacting the cell with an effective amount of a compound described in any one of Appendices 1 to 4, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described in any one of Appendices 5 to 8. [Appendix 11] 11. The method of claim 9 or 10, wherein the cell is a cancer cell. [Appendix 12] 12. The method of claim 11, wherein the cancer is a malignant tumor, a rhabdoid tumor, a CD8+ T-cell lymphoma, an endometrial cancer, an ovarian cancer, a bladder cancer, a gastric cancer, a pancreatic cancer, an esophageal cancer, a prostate cancer, a renal cell carcinoma, a melanoma, a colorectal cancer, a sarcoma, a non-small cell lung cancer, a gastric cancer, or a breast cancer. [Appendix 13] 13. The method of claim 12, wherein the cancer is a sarcoma. [Appendix 14] 14. The method of claim 13, wherein the sarcoma is soft tissue sarcoma, synovial sarcoma, Ewing's sarcoma, osteosarcoma, rhabdomyosarcoma, adult fibrosarcoma, alveolar soft part sarcoma, angiosarcoma, clear cell sarcoma, desmoplastic small round cell tumor, epithelioid sarcoma, fibromyxoid sarcoma, gastrointestinal stromal tumor, Kaposi's sarcoma, liposarcoma, leiomyosarcoma, malignant mesenchymoma, malignant peripheral nerve sheath tumor, myxofibrosarcoma, or low-grade rhabdomyosarcoma. [Appendix 15] 15. The method of claim 14, wherein the sarcoma is synovial sarcoma. [Appendix 16] 15. The method of claim 14, wherein the sarcoma is rhabdomyosarcoma. [Appendix 17] A method of treating a BAF complex-associated disorder in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to any one of appendices 1-4, or a pharma- ceutical composition according to any one of appendices 5-8. [Appendix 18] A method for treating an SS18-SSX fusion protein associated disorder in a subject in need thereof, comprising administering to the subject an effective amount of a compound described in any one of Appendices 1 to 4, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described in any one of Appendices 5 to 8. [Appendix 19] A method for treating a BRD9-related disorder in a subject in need thereof, comprising administering to the subject an effective amount of a compound described in any one of Appendices 1-4, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described in any one of Appendices 5-8. [Appendix 20] 20. The method of any one of claims 17 to 19, wherein the disorder is cancer. [Appendix 21] A method of treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to any one of appendices 1-4, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition according to any one of appendices 5-8. [Appendix 22] 22. The method of claim 20 or 21, wherein the cancer is a malignant tumor, a rhabdoid tumor, a CD8+ T-cell lymphoma, an endometrial cancer, an ovarian cancer, a bladder cancer, a gastric cancer, a pancreatic cancer, an esophageal cancer, a prostate cancer, a renal cell carcinoma, a melanoma, a colorectal cancer, a sarcoma, a non-small cell lung cancer, a gastric cancer, or a breast cancer. [Appendix 23] 23. The method of claim 22, wherein the cancer is a sarcoma. [Appendix 24] 24. The method of claim 23, wherein the sarcoma is soft tissue sarcoma, synovial sarcoma, Ewing's sarcoma, osteosarcoma, rhabdomyosarcoma, adult fibrosarcoma, alveolar soft part sarcoma, angiosarcoma, clear cell sarcoma, desmoplastic small round cell tumor, epithelioid sarcoma, fibromyxoid sarcoma, gastrointestinal stromal tumor, Kaposi's sarcoma, liposarcoma, leiomyosarcoma, malignant mesenchymoma, malignant peripheral nerve sheath tumor, myxofibrosarcoma, or low-grade rhabdomyosarcoma. [Appendix 25] 25. The method of claim 24, wherein the sarcoma is synovial sarcoma. [Appendix 26] 25. The method of claim 24, wherein the sarcoma is rhabdomyosarcoma. [Appendix 27] 23. The method of claim 12 or 22, wherein the cancer is prostate cancer. [Appendix 28] 22. The method of claim 11, 20, or 21, wherein the cancer is acute myeloid leukemia. [Appendix 29] 22. The method of claim 11, 20, or 21, wherein the cancer is a BRCA mutant cancer. [Appendix 30] 20. The method of any one of appendix 17 to 19, wherein the disorder is an infectious disease. [Appendix 31] A method of treating an infectious disease in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to any one of appendices 1 to 4, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition according to any one of appendices 5 to 8. [Appendix 32] 32. The method of claim 30 or 31, wherein the infection is a viral infection. [Appendix 33] 33. The method of claim 32, wherein the viral infection is an infection caused by a virus of the family Retroviridae, Hepadnaviridae, Flaviviridae, Adenoviridae, Herpesviridae, Papillomaviridae, Parvoviridae, Polyomaviridae, Paramyxoviridae, or Togaviridae. [Appendix 34] 34. The method of claim 32 or 33, wherein the viral infection is Bifidobacterium coffinii, neurofibromatosis, or multiple meningiomas.

Claims

1. A compound having the structure: 【Chemistry 1】 or a pharma- ceutically acceptable salt thereof.

2. The compound has the following structure: 【Chemistry 2】 2. The compound of claim 1, having the formula:

3. The compound has the following structure: 【Chemistry 3】 2. The compound of claim 1, having the formula:

4. A compound having the structure: 【Chemistry 4】 or a pharma- ceutically acceptable salt thereof and a pharma- ceutically acceptable excipient.

5. The pharmaceutical composition comprises a compound having the structure: 【Chemistry 5】 5. The pharmaceutical composition of claim 4, comprising:

6. The pharmaceutical composition comprises a compound having the structure: 【Chemistry 6】 5. The pharmaceutical composition of claim 4, comprising:

7. Use of a compound according to any one of claims 1 to 3, or a pharmaceutical composition according to any one of claims 4 to 6, for the manufacture of a medicament for the treatment of cancer in a subject in need thereof.

8. 8. The use of claim 7, wherein the cancer is a malignant tumor, a rhabdoid tumor, a CD8+ T-cell lymphoma, an endometrial cancer, an ovarian cancer, a bladder cancer, a gastric cancer, a pancreatic cancer, an esophageal cancer, a prostate cancer, a renal cell carcinoma, a melanoma, a colorectal cancer, a sarcoma, a non-small cell lung cancer, or a breast cancer.

9. 9. The use of claim 8, wherein the sarcoma is soft tissue sarcoma, synovial sarcoma, Ewing's sarcoma, osteosarcoma, rhabdomyosarcoma, adult fibrosarcoma, alveolar soft part sarcoma, angiosarcoma, clear cell sarcoma, desmoplastic small round cell tumor, epithelioid sarcoma, fibromyxoid sarcoma, gastrointestinal stromal tumor, Kaposi's sarcoma, liposarcoma, leiomyosarcoma, malignant mesenchymoma, malignant peripheral nerve sheath tumor, myxofibrosarcoma, or low-grade rhabdomyosarcoma.

10. The use according to claim 9, wherein the sarcoma is a synovial sarcoma.

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