PERK inhibitor HC-5404 in combination with an anti-PD-1 antibody and / or an anti-angiogenic agent for use in the treatment of cancer

Abstract

Provided herein are methods for treating cancer (e.g., solid tumors) in a subject in need of such treatment. The present disclosure also provides methods for activating the immune system of a subject in need of such activation (e.g., a subject with cancer). The methods described herein generally involve administering to the subject an effective amount of a compound of formula (I): [Case 1] JPEG2025533802000030.jpg28170 or a pharmaceutically acceptable salt thereof, and immunotherapy (e.g., anti-PD-1 antibody) and / or anti-angiogenic agent (e.g., VEGFR-TKI).

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to U.S. Provisional Patent Application Nos. 63 / 443,344, filed February 3, 2023, 63 / 455,911, filed March 30, 2023, and 63 / 413,116, filed October 4, 2022, the contents of each of which are incorporated by reference in their entirety into this specification. [Background technology]

[0002] background Cancer is the leading cause of death worldwide, accounting for nearly 10 million deaths in 2020 (World Health Organization). Targeted therapy and immunotherapy have dramatically improved prognosis, paving the way for cancer treatment. However, tumor recurrence, drug resistance, and drug intolerance remain significant challenges in cancer management (Wang et al., “Drug resistance and combating drug resistance in cancer,” Cancer Drug Resistance, 2019, 2(2): 141-160; Chakraborty et al., “The difficulties in cancer treatment,” Cancer Medical Science, 2012, 6: ed. 16). Cancer manifests as widespread proliferation of tumor cells, which requires the proper functioning of the protein machinery (Waczak et al., 2019). The endoplasmic reticulum (ER) regulates the folding, synthesis, and maturation of proteins required by tumor cells, playing a crucial role in maintaining tumor homeostasis (Yoo et al., 2017). The accumulation of unfolded proteins in the ER lumen activates three major pathways: 1) PERK, 2) inositol-requiring enzyme 1α (IRE1α), and 3) activating transcription factor 6 (ATF6)—collectively known as the unfolded protein response (UPR)—which enable cells to correct this stress and survive (Walter and Ron, 2011; Urra et al., 2016).Evidence suggests that in various types of cancer, UPR activation is a mechanism that allows tumor cells to adapt to the demands on the ER and oxidative conditions imposed by conditions inherent to their biology, such as increased translational load caused by the loss of oncogenes and tumor suppressor genes, and by environmental conditions commonly found in the tumor / metastatic microenvironment, such as hypoxia, among other signals (Blais et al., 2004; Chevet et al., 2015; Tameire et al., 2015; Hart et al., 2012; Martin-Perez et al., 2014; Rajasekhar and Holland, 2004; Rajasekhar et al., 2003; Rojo et al., 2007; Sequeira et al., 2009). Oncogene-activated pathways increase ER client protein load through activation of mTOR signaling and translation initiation (Tameire et al., 2015; Hart et al., 2012; Ozcan et al., 2008). Other studies have further demonstrated that the UPR pathway contributes to adaptation to hypoxia and microenvironmental stress (Blais et al., 2004; Bi et al., 2005; Chen et al., 2014; Romero-Ramirez et al., 2009; Rouschop et al., 2010; Schewe et al., 2008; Ye et al., 2010), suggesting that UPR activation may enable adaptation to a changing environment.

[0003] As a component of the UPR pathway, PERK is involved in tumorigenesis-promoting processes, making it an attractive target for anticancer therapy. More specifically, PERK phosphorylates eukaryotic translation initiation factor 2α (eIF2α), nuclear factor (erythroid-derived 2)-like 2 (Nrf2), and forkhead box protein O1 (FOXO) proteins. Phosphorylation of eIF2α inhibits general protein synthesis, reducing protein load while inducing targeted translation of uORFs containing mRNAs such as ATF4. ATF4 then initiates a gene expression program consisting of, among others, antioxidant, chaperone, and autophagy genes aimed at restoring protein homeostasis and inducing cell survival in the context of ER stress. Cancer cells, by definition, are characterized by cellular activities and microenvironmental factors that contribute to the activation of the UPR (including oncogenic protein synthesis, hypoxic environments, dissemination and residence in different microenvironments, etc.), and in addition, the actions of many therapeutic agents further contribute to the activation of the UPR in cells that are not eliminated by therapy. The UPR pathway, including PERK, has not been a frequently investigated pathway / target for cancer therapy. Summary of the Invention [Problem to be solved by the invention]

[0004]

[0004] Thus, there remains an unmet need to develop novel therapeutic strategies that utilize modulation of the PERK pathway for the treatment of various cancers (eg, solid tumors). [Means for solving the problem]

[0005] overview In one aspect, provided herein is a method of activating the immune system of a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formula (I): [ka] or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of an anti-PD-1 antibody.

[0006] In another aspect, provided herein is a method of activating the immune system of a subject having cancer, comprising administering to the subject a therapeutically effective amount of a compound of formula (I): [ka] or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of an anti-PD-1 antibody.

[0007] In certain embodiments, the method inhibits the activity of myeloid-derived suppressor cells (MDSCs). In certain embodiments, the method increases the infiltration of T cells and NK cells into tumors.

[0008]

[0008] In certain embodiments, the tumor is selected from the group consisting of bladder cancer, head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma and neuroendocrine carcinoma.

[0009] In certain embodiments, the method increases the frequency of dendritic cells in the draining lymph node.

[0010] In another aspect, provided herein is a method of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formula (I): [ka] or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of an anti-PD-1 antibody.

[0011] In certain embodiments, the cancer is a solid tumor. In certain embodiments, the cancer is selected from the group consisting of bladder cancer, head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma.

[0012] In certain embodiments, the cancer is a liquid tumor. In certain embodiments, the cancer is selected from the group consisting of classical Hodgkin's lymphoma, primary thymic-mediastinal lymphoma, multiple myeloma, and B-cell malignancies (e.g., non-Hodgkin's lymphoma or chronic lymphocytic leukemia).

[0013] In certain embodiments, the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, and cemiplimab. In certain embodiments, the anti-PD-1 antibody is pembrolizumab. In certain embodiments, the anti-PD-1 antibody is nivolumab.

[0014] In certain embodiments, a therapeutically effective amount of an anti-PD-1 antibody is administered once daily. In certain embodiments, a therapeutically effective amount of an anti-PD-1 antibody is administered twice daily.

[0015] In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody are administered simultaneously. In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody are administered sequentially.

[0016] In certain embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered in an amount of about 22 mg to about 451 mg twice daily. In certain embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered in an amount of about 177 mg to about 900 mg twice daily.

[0017] In certain embodiments, a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof is administered orally. In certain embodiments, a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in capsule form.

[0018] In certain embodiments, the method increases tumor cell expression of type 1 interferon receptor (INFAR1) in the subject compared to a subject receiving a placebo, a therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of an anti-PD-1 antibody. In certain embodiments, the method increases peripheral blood monocyte surface expression of INFAR1 in the subject compared to a subject receiving a placebo, a therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of an anti-PD-1 antibody. In certain embodiments, the method increases tumor cell expression of calreticulin in the subject compared to a subject receiving a placebo, a therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of an anti-PD-1 antibody. In certain embodiments, the method increases polymorphonuclear myeloid-derived suppressor cell (MDSC) and / or tumor-associated macrophage (TAM) expression of INFAR1 in the subject compared to subjects receiving a placebo, a therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of an anti-PD-1 antibody. In certain embodiments, the method increases TAM expression of PD-L1 in the subject compared to subjects receiving a placebo, a therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of an anti-PD-1 antibody. In certain embodiments, the method increases tumor infiltration of CD8 T cells and / or NK cells in the subject compared to subjects receiving a placebo, a therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of an anti-PD-1 antibody. In certain embodiments, the method increases CD69 expression on T cells in tumor-draining lymph nodes in the subject, compared to a subject receiving a placebo, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of an anti-PD-1 antibody. In certain embodiments, the method reduces the suppressive activity of MDSCs in the subject. In certain embodiments, the method increases dendritic cell frequency in tumor-draining lymph nodes in the subject. In certain embodiments, the method increases Ki67, granzyme B (GzmB), and / or memory phenotype markers on CD4 T cells in the subject.

[0019]

[0019] In certain embodiments, the compound of formula (I) is administered as a pharmaceutically acceptable salt. In certain embodiments, the pharmaceutically acceptable salt is the hemifumarate salt.

[0020]

[0020] In another aspect, provided herein is a method of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formula (I): [ka] or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of an anti-angiogenic agent.

[0021]

[0021] In another aspect, provided herein is a method of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formula (I): [ka] or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-angiogenic agent, wherein the subject is being treated with an anti-cancer therapy.

[0022] In some embodiments, the subject is resistant or has acquired resistance to the anti-cancer therapy.

[0023]

[0023] In some embodiments, the anti-cancer therapy is selected from the group consisting of administration of immunotherapeutic agents, chemotherapeutic agents, growth inhibitors, cytotoxic agents, anti-angiogenic agents or combinations thereof, radiation therapy, surgery and combinations thereof.

[0024] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-angiogenic agent are administered simultaneously.

[0025] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-angiogenic agent are administered sequentially.

[0026]

[0026] In some embodiments, a therapeutically effective amount of an anti-angiogenic agent is administered to a subject daily.

[0027] In some embodiments, a therapeutically effective amount of an anti-angiogenic agent is administered to a subject once daily.

[0028] In some embodiments, a therapeutically effective amount of an antiangiogenic agent is administered to a subject twice daily.

[0029]

[0029] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-angiogenic agent reduces p-PERK levels in a subject compared to administering a therapeutically effective amount of the anti-angiogenic agent alone.

[0030]

[0030] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-angiogenic agent increases ATF4 levels in a subject compared to administering a therapeutically effective amount of the anti-angiogenic agent alone.

[0031]

[0031] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-angiogenic agent modulates activation of the unfolded protein response (UPR) compared to administering a therapeutically effective amount of the anti-angiogenic agent alone.

[0032] In another aspect, provided herein is a method of treating cancer in a subject receiving a therapeutically effective amount of anti-angiogenic therapy, comprising administering to the subject a therapeutically effective amount of a compound of formula (I): [ka] or a pharmaceutically acceptable salt thereof.

[0033] In some embodiments, the subject is resistant or has acquired resistance to antiangiogenic therapy.

[0034]

[0034] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof reduces p-PERK levels in a subject compared to administering a therapeutically effective amount of an anti-angiogenic agent alone.

[0035]

[0035] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof increases ATF4 levels in a subject compared to administering a therapeutically effective amount of an anti-angiogenic agent alone.

[0036]

[0036] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof modulates activation of the unfolded protein response (UPR) compared to administering a therapeutically effective amount of an anti-angiogenic agent alone.

[0037]

[0037] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof reduces tumor vasculature in a subject compared to administering a therapeutically effective amount of an anti-angiogenic agent alone.

[0038] In some embodiments, the anti-angiogenic agent is a vascular endothelial growth factor receptor (VEGFR) modulator.

[0039] In some embodiments, the anti-angiogenic agent is a vascular endothelial growth factor (VEGF)-targeting antibody.

[0040] In some embodiments, the antiangiogenic agent is a tyrosine kinase inhibitor (TKI).

[0041] In some embodiments, the antiangiogenic agent is a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI).

[0042]

[0042] In some embodiments, the antiangiogenic agent is selected from the group consisting of apatinib, axitinib, brivanib, cabozantinib, canertinib, cediranib, dasatinib, erlotinib, gefitinib, leflunomide, lenvatinib, motesanib, nilotinib, nintedanib, pazopanib, regorafenib, semaxinib, sorafenib, sunitinib, tivozanib, vandetanib, vatalanib, XL0192, bevacizumab, ramucirumab and pharmaceutically acceptable salts or biosimilars thereof.

[0043]

[0043] In some embodiments, the antiangiogenic agent is selected from the group consisting of bevacizumab, ramucirumab and biosimilars thereof.

[0044]

[0044] In some embodiments, the antiangiogenic agent is selected from the group consisting of apatinib, axitinib, brivanib, cabozantinib, canertinib, cediranib, dasatinib, erlotinib, gefitinib, leflunomide, lenvatinib, motesanib, nilotinib, nintedanib, pazopanib, regorafenib, semaxinib, sorafenib, sunitinib, tivozanib, vandetanib, vatalanib, XL0192 and pharmaceutically acceptable salts thereof.

[0045] In some embodiments, the cancer is a metastatic cancer or a locally advanced cancer.

[0046] In some embodiments, the cancer is a solid tumor.

[0047]

[0047] In some embodiments, the cancer is selected from the group consisting of bladder cancer, breast cancer, carcinoma, cervical cancer, colorectal cancer, gastric cancer, hepatocellular carcinoma, renal cancer, lung cancer, neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer and thyroid cancer.

[0048] In some embodiments, the cancer is selected from the group consisting of breast cancer, gastric cancer, renal cancer and lung cancer.

[0049]

[0049] In some embodiments, the breast cancer is triple-negative breast cancer or metastatic breast cancer, the carcinoma is carcinoma of unknown primary (CUP), endometrial carcinoma, head and neck squamous cell carcinoma, Merkel cell carcinoma, squamous cell carcinoma or urothelial carcinoma, the gastric cancer is adenocarcinoma or gastrointestinal stromal tumor, the renal cancer is renal cell carcinoma (RCC), or the lung cancer is small cell lung cancer (SCLC) or non-small cell lung carcinoma.

[0050]

[0050] In some embodiments, the breast cancer is triple-negative breast cancer or metastatic breast cancer.

[0051] In some embodiments, the carcinoma is carcinoma of unknown primary (CUP), endometrial carcinoma, head and neck squamous cell carcinoma, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma.

[0052]

[0052] In some embodiments, the gastric cancer is an adenocarcinoma or a gastrointestinal stromal tumor.

[0053] In some embodiments, the renal cancer is renal cell carcinoma (RCC).

[0054] In some embodiments, the lung cancer is small cell lung cancer (SCLC) or non-small cell lung cancer.

[0055]

[0055] In some embodiments, the renal cell carcinoma is clear cell renal cell carcinoma (ccRCC).

[0056]

[0056] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof comprises administering to a subject about 22 mg to about 798 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0057]

[0057] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject about 22 mg to about 798 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0058]

[0058] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject from about 22 mg to about 798 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof daily.

[0059]

[0059] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject about 22 mg to about 798 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof once daily.

[0060]

[0060] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject about 22 mg to about 798 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof twice daily.

[0061]

[0061] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof comprises administering to a subject about 44 mg to about 177 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0062]

[0062] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject about 44 mg to about 177 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0063]

[0063] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject from about 44 mg to about 177 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof daily.

[0064]

[0064] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject about 44 mg to about 177 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof once daily.

[0065]

[0065] In some embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject about 44 mg to about 177 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof twice daily.

[0066] In some embodiments, the subject is in a fasting state.

[0067] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered with food.

[0068] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered about 30 minutes to about 1 hour after a meal.

[0069] In some embodiments, a therapeutically effective amount of a compound of Formula (I) is administered as a pharmaceutically acceptable salt.

[0070] In some embodiments, the pharmaceutically acceptable salt is a hemifumarate salt.

[0071]

[0071] In some embodiments, the method further comprises administering to the subject a therapeutically effective amount of immunotherapy.

[0072]

[0072] In certain aspects, provided herein are methods of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formula (I): [ka] or a pharmaceutically acceptable salt thereof, a therapeutically effective amount of an anti-angiogenic agent, and a therapeutically effective amount of an immunotherapy.

[0073] In another aspect, provided herein is a method of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formula (I): [ka] or a pharmaceutically acceptable salt thereof, a therapeutically effective amount of an anti-angiogenic agent, and a therapeutically effective amount of an immunotherapy, wherein the subject is being treated with an anti-cancer therapy.

[0074] In some embodiments, the subject is resistant or has acquired resistance to the anti-cancer therapy.

[0075]

[0075] In some embodiments, the anti-cancer therapy is selected from the group consisting of administration of immunotherapeutic agents, chemotherapeutic agents, growth inhibitors, cytotoxic agents, anti-angiogenic agents or combinations thereof, radiation therapy, surgery and combinations thereof.

[0076] In another aspect, provided herein is a method of treating cancer in a subject receiving a therapeutically effective amount of anti-angiogenic therapy, comprising administering to the subject a therapeutically effective amount of a compound of formula (I): [ka] or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of immunotherapy.

[0077]

[0077] In some embodiments, the immunotherapy is an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody.

[0078]

[0078] In some embodiments, the immunotherapy is a therapeutically effective amount of an anti-PD-1 antibody.

[0079]

[0079] In some embodiments, the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, and cemiplimab.

[0080]

[0080] In some embodiments, the anti-PD-1 antibody is pembrolizumab.

[0081]

[0081] In some embodiments, the anti-PD-1 antibody is nivolumab.

[0082]

[0082] In some embodiments, a therapeutically effective amount of immunotherapy is administered daily.

[0083]

[0083] In some embodiments, a therapeutically effective amount of immunotherapy is administered once daily.

[0084]

[0084] In some embodiments, a therapeutically effective amount of immunotherapy is administered twice daily.

[0085] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapy are administered simultaneously.

[0086] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapy are administered sequentially.

[0087]

[0087] In some embodiments, a therapeutically effective amount of an anti-angiogenic agent and a therapeutically effective amount of an immunotherapy are administered simultaneously.

[0088]

[0088] In some embodiments, the therapeutically effective amount of an anti-angiogenic agent and the therapeutically effective amount of an immunotherapy are administered sequentially. [Brief explanation of the drawings]

[0089] Brief description of the diagram [Figure 1A]

[0089] Figure 10 is a graph showing the effect of single agent 10 mg / kg HC-5404 twice daily or 30 mg / kg HC-5404 twice daily on tumor volume compared to vehicle in a renal cell carcinoma model. [Figure 1B]

[0090] 1 is a graph showing the effect of single agent 30 mg / kg HC-5404 twice daily compared to vehicle on tumor volume in a gastric cancer model. [Figure 2A]

[0091] 1 is a scheme showing the study design of the MB49 study described in Example 2. [Figure 2B]

[0092] 10 mg / kg HC-5404-FU orally twice daily, 30 mg / kg HC-5404-FU orally twice daily, 5 mg / kg anti-PD-1 antibody intraperitoneally every other week, 3 mg / kg HC-5404-FU orally twice daily + 5 mg / kg anti-PD-1 antibody intraperitoneally every other week, 10 mg / kg HC-5404-FU orally twice daily + 5 mg / kg anti-PD-1 antibody intraperitoneally every other week, or 30 mg / kg HC-5404-FU orally twice daily + 5 mg / kg anti-PD-1 antibody intraperitoneally every other week after inoculation with MB49 cells. [Figure 2C]

[0093] 1 is a graph showing tumor volume after MB49 cell inoculation in MB49 tumor-bearing mice treated with vehicle, 30 mg / kg HC-5404-FU orally twice daily and 5 mg / kg anti-PD-1 antibody intraperitoneally every other week, or 30 mg / kg HC-5404-FU orally twice daily plus 5 mg / kg anti-PD-1 antibody intraperitoneally every other week. [Figure 2D]

[0094] 2C is a graph showing the tumor volume and number of responders for individual subjects after cell inoculation in the vehicle-treated group depicted in FIG. 2C. [Figure 2E]

[0095] 2C is a graph showing tumor volumes and number of responders for individual subjects after cell inoculation in the 30 mg / kg HC-5404-FU treatment group depicted in FIG. 2C. [Figure 2F]

[0096] 2C is a graph showing the tumor volume and number of response cases for individual subjects after cell inoculation in the anti-PD-1 antibody treatment group. [Figure 2G]

[0097] 2C is a graph showing the tumor volume and number of responders for individual subjects after cell inoculation in the 30 mg / kg HC-5404-FU + anti-PD-1 antibody treatment group. [Figure 3A]

[0098] FIG. 2C is a graph showing IFNAR1 expression on M1 macrophages from the treatment groups depicted in FIG. 2B at 14 days post-inoculation (7 days post-treatment) as determined by flow cytometry. [Figure 3B]

[0099] FIG. 2C is a graph showing IFNAR1 expression on M2 macrophages from the treatment groups depicted in FIG. 2B at 14 days post-inoculation (7 days post-treatment) as determined by flow cytometry. [Figure 3C]

[0100] 2C is a graph showing IFNAR1 expression on polymorphonuclear MDSCs from the treatment groups depicted in FIG. 2B at 14 days post-inoculation (7 days post-treatment) as determined by flow cytometry. [Figure 3D]

[0101] FIG. 2C is a graph showing IFNAR1 expression on tumor cells from the treatment groups depicted in FIG. 2B at 14 days post-inoculation (7 days post-treatment) as determined by flow cytometry. [Figure 3E]

[0102] Figure 2C is a graph showing PD-L1 expression on macrophages from the treatment groups depicted in Figure 2B at 14 days post-inoculation (7 days post-treatment) as determined by flow cytometry. [Figure 4A]

[0103] FIG. 1 shows proteins involved in immunogenic cell death in cancer therapy. [Figure 4B]

[0104] FIG. 2C is a graph showing surface calreticulin expression on tumor cells from the treatment groups depicted in FIG. 2B at 14 days post-inoculation (7 days post-treatment) as determined by flow cytometry. [Figure 4C]

[0105] Image comparison of 786-O cells from a xenograft model treated with HC-5404 or vehicle, showing HMGB1 in the cytoplasm as detected by immunohistochemistry. [Figure 4D]

[0106] 1 is a graph showing HMGB1 cytoplasmic expression induced by 30 mg / kg HC-5404 treatment compared to vehicle in a 786-O xenograft model 15 days after treatment. [Figure 5A]

[0107] FIG. 2C is a graph showing the frequency of tumor-infiltrating CD8 T cells in the treatment groups depicted in FIG. 2B at day 7 post-treatment, as determined by flow cytometry. [Figure 5B]

[0108] FIG. 2C is a graph showing the frequency of tumor-infiltrating CD8 NK cells in the treatment groups depicted in FIG. 2B at day 7 post-treatment, as determined by flow cytometry. [Figure 5C]

[0109] FIG. 2C is a graph showing the frequency of CD69 expression on CD4 T cells from lymph nodes of the treatment groups depicted in FIG. 2B at day 7 post-treatment, as determined by flow cytometry. [Figure 5D]

[0110] FIG. 2C is a graph showing the frequency of CD69 expression on CD8 T cells from lymph nodes of the treatment groups depicted in FIG. 2B at day 7 post-treatment, as determined by flow cytometry. [Figure 6A]

[0111] FIG. 2C is a graph showing Ki67 expression on CD4 T cells in lymph nodes of the treatment groups depicted in FIG. 2B at 14 days post-treatment, as determined by flow cytometry. [Figure 6B]

[0112] FIG. 2C is a graph showing EOMES expression on CD4 T cells in lymph nodes of the treatment groups depicted in FIG. 2B at 14 days post-treatment, as determined by flow cytometry. [Figure 6C]

[0113] FIG. 2C is a graph showing granzyme B expression on CD4 T cells in lymph nodes of the treatment groups depicted in FIG. 2B at 14 days post-treatment, as determined by flow cytometry. [Figure 6D]

[0114] FIG. 2C is a graph showing effector memory populations of CD4 T cells in lymph nodes of the treatment groups depicted in FIG. 2B at 14 days post-treatment, as determined by flow cytometry. [Figure 6E]

[0115] FIG. 2C is a graph showing TCF1 expression on CD4 T cells in lymph nodes of the treatment groups depicted in FIG. 2B at 14 days post-treatment, as determined by flow cytometry. [Figure 7A]

[0116] 2C is a graph showing total dendritic cells in lymph nodes of the treatment groups depicted in FIG. 2B at 14 days post-treatment, as determined by flow cytometry. [Figure 7B]

[0117] 2C is a graph showing XCR1+ cDC1 cells in lymph nodes of the treatment groups depicted in FIG. 2B at 14 days post-treatment, as determined by flow cytometry. [Figure 7C]

[0118] 2C is a graph showing cDC2 cells in lymph nodes of the treatment groups depicted in FIG. 2B at 14 days post-treatment, as determined by flow cytometry. [Figure 8]

[0119] 2B is a graph comparing the expression of IFNAR1 on monocytes from whole blood of the vehicle, 10 mg / kg HC-5404, and 30 mg / kg HC-5404 treatment groups depicted in FIG. 2B on days 7, 11, and 14 after treatment, as determined by flow cytometry. [Figure 9]

[0120] 2B is a heatmap showing the expression of myeloid (M1 and M2 related), lymphoid (T cell related), and interferon-activated genes (ISGs) in the vehicle, 10 mg / kg HC-5404, and 30 mg / kg HC-5404 treatment groups depicted in FIG. 2B at day 7 post-treatment. [Figure 10A]

[0121] 1 shows the results of a T cell suppression assay performed using human CD3 T cells co-cultured with human MDSCs and treated with vehicle, 500 nM HC-5404, or 750 nM HC-5404 (1:1 MDSC:T cell ratio). [Figure 10B]

[0122] 1 is a scheme showing the preparation of differentiated human MDSCs from umbilical cord blood. [Figure 10C]

[0123] 10B is a histogram showing the results of the T cell suppression assay depicted in FIG. 10A. [Figure 11A]

[0124] 1 shows the results of a T cell suppression assay performed using murine CD8 T cells co-cultured with MDSCs and treated with RPMI medium, LSLG, or LSLG+500 nM HC-5404 (MDSC:T cell ratio of 8:1). [Figure 11B]

[0125] 1 is a scheme showing the preparation of murine MDSCs from the bone marrow of non-tumor-bearing mice. [Figure 11C]

[0126] FIG. 11B is a set of histograms showing the results of the suppression assay depicted in FIG. 11A and a similar assay performed on mouse CD4 T cells. [Figure 12]

[0127] 1 is a graph showing the FRET-based biochemical assay used to evaluate HC-5404 inhibition of the ISR kinase GCN2 over a concentration series ranging from 0.2 nM to 10 μM. [Figure 13]

[0128] 1 is a graph showing the FRET-based biochemical assay used to evaluate HC-5404 inhibition of the ISR kinase PERK over a concentration series ranging from 0.2 nM to 10 μM. [Figure 14]

[0129] 1 is a graph showing the FRET-based biochemical assay used to assess HC-5404 inhibition of the ISR kinase HRI over a concentration series ranging from 0.2 nM to 10 μM. [Figure 15]

[0130] 1 is a graph showing the FRET-based biochemical assay used to evaluate HC-5404 inhibition of the ISR kinase GCN2 over a concentration series ranging from 0.2 nM to 10 μM. [Figure 16]

[0131] 1 is a table demonstrating that HC-5404 is PERK-selective over four closely related ISR kinases. [Figure 17]

[0132] Biochemical TreeSpot kinome panel assay demonstrating the selectivity of HC-5404 (100 nM, 1000 nM, 10,000 nM) against over 400 kinases. PERK was not included in the panel, and no interaction was observed when HC-5404 was assayed at 100 nM. [Figure 18]

[0133] Figure 1 shows free drug levels of HC-5404 in plasma after a single oral dose. Plasma samples were collected over 24 hours and quantified by LC-MS / MS. [Figure 19]

[0134] 1 shows the PK / PD relationship between plasma exposure and pPERK levels in the mouse pancreas after oral administration of HC-5404 at 30 mg / kg. [Figure 20]

[0135] 1 shows the PK / PD relationship between plasma exposure and pPERK levels in the mouse pancreas after oral administration of HC-5404 at 100 mg / kg. [Figure 21]

[0136] PD effect of HC-5404 on pPERK in 786-O tumors sampled 1, 4, 8, 12 hours after the final dose after 15 days of BID dosing is shown. [Figure 22]

[0137]

[0023] Figure 1 is a graph showing tumor volumes for 786-O xenografts treated for 28 days with HC-5404 at multiple dose levels and treatment regimens. Values ​​represent mean tumor volume ± SEM. [Figure 23]

[0138] 1 is a graph showing tumor volumes in mice harboring subcutaneous CAPAN-2 pancreatic tumor xenografts treated with HC-5404 at 30 mg / kg and 100 mg / kg PO, BID for 48 days. Values ​​represent mean tumor volume ± SEM. [Figure 24]

[0139] Mouse pancreatic sections after 3 weeks of treatment with HC-5404 at either 30 mg / kg PO BID or 100 mg / kg PO BID. The right pancreas was treated with 100 mg / kg PO BID followed by a 2-week washout period to demonstrate whether this effect is reversible. [Figure 25]

[0140] 1 is a graph demonstrating pPERK / PERK protein ratios from 786-O xenografts treated with sunitinib (40 mg / kg; PO; QD) for 1 or 2 weeks. [Figure 26]

[0141] A, D, G) 786-O tumor xenografts treated with cabozantinib (15, 30, 45 mg / kg; PO; QD), lenvatinib (5 and 10 mg / kg; PO; QD), or axitinib (15 and 30 mg / kg; PO; BID) for 21 days. B, E, H) After 21 days of treatment, relative pPERK to total PERK was assessed from tumor samples collected at the end of the study. Differences between protein abundances were compared by ANOVA statistical analysis. Bars highlight significant differences between groups, including p-values. C, F, I) Mouse body weights were measured twice weekly throughout the entire course of treatment. [Figure 27]

[0142] Simple Western protein analysis of pPERK and PERK in VEGFR-TKI treated 786-O xenografts in panel B, sampled after 7 days of treatment. [Figure 28]

[0143] FIG. 1 is a graph demonstrating tumor volume of 786-O tumor xenografts treated for 28 days with HC-5404 (30 mg / kg; BID), sunitinib (40 mg / kg; QD), lenvatinib (10 mg / kg; BID), axitinib (30 mg / kg; BID), or cabozantinib (30 mg / kg; QD) and combinations as indicated. [Figure 29]

[0144] It is demonstrated that HC-5404 induces the accumulation of ASNS, CBS and CTH. [Figure 30]

[0145] Shown are pPERK and total PERK abundance assessed by Simple Western in RCC tumor xenografts treated for 7 days with HC-5404 (30 mg / kg; PO; BID), sunitinib (20 or 40 mg / kg; PO; QD as indicated), or their combination. [Figure 31]

[0146] A498 RCC tumor xenografts treated with HC-5404 (30 mg / kg; PO; BID) and sunitinib (20 mg / kg; PO; QD) for 30 days are shown. [Figure 32]

[0147] Caki-1 RCC tumor xenografts treated with HC-5404 (30 mg / kg; PO; BID) and sunitinib (20 mg / kg; PO; QD) for 30 days are shown. [Figure 33]

[0148] Growth curves of a representative RCC PDX model over 28 days of treatment with either axitinib (30 mg / kg; PO; BID), HC-5404 (30 mg / kg; PO; BID), or their combination. [Figure 34]

[0149] Growth curves of a representative RCC PDX model over 28 days of treatment with either axitinib (30 mg / kg; PO; BID), HC-5404 (30 mg / kg; PO; BID), or their combination. [Figure 35]

[0150] 1 is a waterfall plot illustrating the relative change in tumor volume of 15 different RCC PDX models after 28 days of treatment with either axitinib (30 mg / kg; PO; BID), HC-5404 (30 mg / kg; PO; BID), or their combination. Models were ranked according to the % tumor volume change in the combination group. Progressive disease (PD) = >30% increase from baseline; stable disease (SD) = <30% increase from baseline and <50% regression; partial response (PR) = >50% regression. Models that are VHL wild-type are indicated with an asterisk. [Figure 36]

[0151] Figure 1 shows the sensitivity of all three RCC xenograft models treated with HC-5404 at 10 and 30 mg / kg PO BID for 28 days. 786-O and A498 are VHL mutant models; Caki-1 is VHL wild-type. [Figure 37]

[0152] RCC tumor models treated with HC-5404 (30 mg / kg; PO; BID) and DC-101 (15 mg / kg; IP; BIW) for 28 days are shown. Mean tumor volume ± SEM is shown. B, D, F) Ratio of pPERK to total PERK for all four treatment groups in three RCC models, as assessed by Simple Western protein analysis system. Note: Low signal-to-noise ratio in the Caki-1 model. [Figure 38]

[0153] IHC images of 786-O xenograft sections stained with antibodies specific for Meca32, CD31, and SMA are shown. Scale bar = 150 μm. [Figure 39]

[0154] Quantification of IHC staining of xenograft sections is shown. Graphs show the percentage of cells positively stained for Meca32+, or positively stained for CD31 in the absence of SMA (%CD31+SMA-), or positively stained for both CD31 and SMA (%CD31+SMA+). Differences between treatment groups were assessed by one-way ANOVA statistical analysis. Significant differences between treatments versus vehicle are indicated by bars and p-values. [Figure 40]

[0155] Shown is 786-O tumor growth over the entire study period. Xenografts progressed in the presence of axitinib (30 mg / kg; PO; BID) for 14 days were re-randomized and crossed over to the indicated treatment group for an additional 28 days. [Figure 41]

[0156] IHC images of xenograft sections stained with antibodies specific for CD31 and SMA are shown. [Figure 42]

[0157] Quantification of the percentage of cells positively stained for SMA, CD31, or Meca32 is shown. Statistical significance between groups was assessed using one-way ANOVA. p values ​​indicated significant differences between treatment and vehicle. Quantification of immunohistochemistry (IHC) staining of FFPE sections from 786-O RCC tumor xenografts treated with HC-5404 (30 mg / kg; PO; BID), axitinib (30 mg / kg; PO; BID), or their combination. Quantification of the % positive cells stained for CD31, a marker of vascular endothelium. [Figure 43]

[0158] Figure 1 shows quantification of IHC staining of tumor sections using antibodies specific for the pericyte markers NG2 and MCAM. Statistical significance between groups was assessed using one-way ANOVA. p values ​​indicated significant differences between treatment groups relative to either baseline or vehicle. Quantification of immunohistochemistry (IHC) staining of FFPE sections from 786-O RCC tumor xenografts treated with HC-5404 (30 mg / kg; PO; BID), axitinib (30 mg / kg; PO; BID), or their combination. Quantification of the percent positive cells stained with antibodies specific for NG2 and MCAM, two pericyte markers. [Figure 44]

[0159] Images and quantification of IHC staining of tumor sections using antibodies specific for the pericyte markers NG2 and MCAM are shown. Statistical significance between groups was assessed using one-way ANOVA. p values ​​indicated significant differences between treatment groups relative to either baseline or vehicle. [Figure 45]

[0160] Tumor growth curves and pPERK / PERK levels relative to total PERK in 786-O tumor xenografts treated with cabozantinib (15, 30, 45 mg / kg; PO; QD) for 21 days are shown. Differences between protein abundances were compared by ANOVA statistical analysis. Bars highlight significant differences between groups, including p-values. [Figure 46]

[0161] Tumor growth curves and pPERK / PERK levels relative to total PERK in 786-O tumor xenografts treated with lenvatinib (5 and 10 mg / kg; PO; QD) for 21 days are shown. Differences between protein abundances were compared by ANOVA statistical analysis. Bars highlight significant differences between groups, including p-values. [Figure 47]

[0162] Figure 1 shows tumor growth curves and pPERK / PERK levels relative to total PERK in 786-O tumor xenografts treated with axitinib (15 and 30 mg / kg; PO; BID) for 21 days. Differences between protein abundances were compared by ANOVA statistical analysis. Bars highlight significant differences between groups, including p-values. [Figure 48]

[0163] 1 shows tumor growth curves of 786-O tumor xenografts treated with lenvatinib (10 mg / kg) administered orally once daily, HC-5404 (30 mg / kg) administered orally twice daily, or a combination thereof. [Figure 49]

[0164] 1 shows tumor growth curves of 786-O tumor xenografts treated with axitinib (30 mg / kg) administered orally twice daily, HC-5404 (30 mg / kg) administered orally twice daily, or a combination thereof. [Figure 50]

[0165] 1 shows tumor growth curves of A498 tumor xenografts treated with oral sunitinib (20 mg / kg) once daily, oral HC-5404 (30 mg / kg) twice daily, or a combination thereof. [Figure 51]

[0166] 1 shows tumor growth curves of A498 tumor xenografts treated with twice-weekly intraperitoneal injections of DC-101 (15 mg / kg), twice-daily oral administration of HC-5404 (30 mg / kg), or a combination thereof. [Figure 52]

[0167]

[0023] Figure 1 shows tumor growth curves for 786-O tumor xenografts treated with twice-daily oral doses of 3, 10, or 30 mg / kg HC-5404 as a single agent or in combination with a once-daily oral dose of 30 mg / kg cabozantinib. Combining 10 or 30 mg / kg HC-5404 with cabozantinib resulted in approximately 66% tumor regression. [Figure 53]

[0168] Body weight curves are shown for 786-O tumor xenografts treated with twice-daily oral doses of 3, 10, or 30 mg / kg HC-5404 as single agents or in combination with a once-daily oral dose of 30 mg / kg cabozantinib. [Figure 54]

[0169] IHC staining of CD31+SMA- cells treated with cabozantinib in combination with HC-5404 is shown. [Figure 55]

[0170] Quantification of IHC staining of CD31+SMA- cells treated with cabozantinib in combination with HC-5404. [Figure 56]

[0171] IHC staining of pericytes treated with cabozantinib in combination with HC-5404 is shown. [Figure 57]

[0172] Quantification of IHC staining of pericytes treated with cabozantinib in combination with HC-5404. Quantification of IHC staining of 786-O RCC tumor xenografts treated with 3, 10, or 30 mg / kg HC-5404 (PO; BID) as a single agent or in combination with cabozantinib (30 mg / kg; PO; QD). [Figure 58]

[0173] 1 shows tumor growth curves for RENCA (VHLwt) RCC tumor xenografts treated with HC-5404, axitinib, anti-PD-1, or a combination thereof. [Figure 59]

[0174] 1 shows weight curves for RENCA (VHLwt) RCC tumor xenografts treated with HC-5404, axitinib, anti-PD-1, or a combination thereof. [Figure 60]

[0175] 1 shows graphs demonstrating the effect of HC-5404 in combination with axitinib on the RENCA RCC model. [Figure 61]

[0176] Graphs demonstrating the effects of HC-5404, axitinib, or HC-5404 in combination with axitinib on the RENCA RCC model are shown. Top left: M1 macrophages (CD11b+, F480+, MHCII+, CD206-) were quantified as a percentage of total F480+ macrophages. Top right: Monocytic MDSCs (CD11b+, Gr-1+, F480-, Ly6C+, Ly6G-) were quantified as a percentage of total CD45+ cells. Bottom left and bottom right: Expression of costimulatory and activation markers CD86 and MHCII on M1 macrophages was measured based on geometric mean fluorescence intensity (MFI). Graphs represent individual animals per treatment group ± SEM. Indicators of significance are based on conventional one-way ANOVA and Tukey's multiple comparison test. [Figure 62]

[0177] Graphs demonstrating the effect of HC-5404, axitinib, or HC-5404 in combination with axitinib on CD4 T cells in tumor-draining lymph nodes are shown. Top left: CD4 T cells expressing the activation marker CD69 were quantified as a percent positive of total CD4 T cells. Top right: Effector memory CD4 T cells (CD4+, CD44+, CD62L-) in the TDLN were quantified as a percentage of total CD4 cells. Center left: TNFα+ CD4 T cells were compared between treatment groups. Center right; bottom left; bottom right: Multiple depletion markers were measured as a percent positive of total CD4 T cells. Graphs represent individual animals ± SD per treatment group. Significance was based on a standard one-way ANOVA and Tukey's multiple comparison test. [Figure 63]

[0178] Demonstrate the effects of HC-5404, axitinib, and HC-5404 in combination with axitinib on myeloid cells in the tumor microenvironment of RENCA RCC tumors. [Figure 64]

[0179] This figure demonstrates the effect of HC-5404, axitinib, and HC-5404 in combination with axitinib on myeloid cells in the tumor microenvironment of RENCA RCC tumors. Fluorescence microscopy is shown for the following markers on macrophages: M2 marker CD206 (top left), and M1 markers CD86 (top right), and PD-L1 (bottom). Values ​​indicate the percentage of macrophages expressing each marker. Points represent individual animals, and error bars indicate SD for each group. Significance is based on a conventional one-way ANOVA and Tukey's multiple comparison test. [Figure 65]

[0180] Demonstrates the effect of HC-5404, axitinib, aPD-1, or a combination of HC-5404 with axitinib and aPD-1 on macrophages in the RENCA RCC model. Macrophages exhibiting an M1-like phenotype (MHCII+, CD206-) (left) or an M2-like phenotype (MHCII-, CD206+) (right) were quantified as a percentage of total F480+ macrophages. Individual animal ± SEM is shown. Significance is based on one-way ANOVA with Dunnett's multiple comparisons. [Figure 66]

[0181] Demonstrates the effect of HC-5404 in combination with axitinib and aPD-1 on the Renca RCC model. [Figure 67]

[0182] Demonstrates the effect of HC-5404, axitinib, aPD-1, and HC-5404 in combination with axitinib and aPD-1 on T cell infiltration. Top left, top right: The frequency (% vs. total CD45+ cells) and cells / gram of CD8 T cells in tumor tissue are shown for the different treatment groups. Bottom left, bottom right: Expression of the stimulatory cytokines IFNγ and TNFα in CD8 T cells was measured by intracellular flow cytometry. Expression is shown as the % positive rate of CD8 T cells. Individual animals ± SD are shown. Significance is based on one-way ANOVA with Dunnett's multiple comparisons. [Figure 68]

[0183] 1 demonstrates the effect of HC-5404 in combination with aPD-1 on myeloid cells in the tumor microenvironment and T cell activation. Tumor volumes were measured over time and are shown for each group (mean ± SEM). [Figure 69]

[0184] This figure demonstrates the effect of HC-5404 in combination with aPD-1 on myeloid cells and T cell activation in the tumor microenvironment in a syngeneic MB49 model. Top (M1 macrophages, M2 macrophages, PMN-MDSC): Expression of type 1 interferon receptor (IFNAR1) on M1-like and M2-like macrophages and MDSC in tumor tissue was assessed by flow cytometry after 7 days of treatment. Expression levels are shown as the geometric mean fluorescence intensity (MFI) of IFNAR1 for each cell type. Center left (CD8+ T cells, NK cells): Tumor-infiltrating CD8+ T cells and NK cells were assessed by flow cytometry after 7 days of treatment. Calculated cells / g of tissue for each group are shown. Center right (In draining lymph nodes: increase in DCs): Dendritic cell (DC) frequency in tumor-draining lymph nodes (TDLN) was measured after 14 days of treatment. Values ​​are shown as the % of total CD45+ cells. Bottom (T cell activation: Ki67, GzmB, TCF1, CD69): CD4 or CD8 T cells in TDLN were assessed for proliferation and activation markers, including Ki67, Granzyme B, TCF1, and CD69. Expression levels are displayed as either % positive cells or MFI for each marker, as indicated. Individual animals ± SD are shown. Significance is based on one-way ANOVA with Dunnett's multiple comparisons. [Figure 70]

[0185] 1 shows the efficacy of HC-5404 on RT4 bladder cells. [Figure 71]

[0186] 1 shows the efficacy of HC-5404 against HCT116 CRC cells. [Figure 72]

[0187] 1 shows the efficacy of HC-5404 on A549 NSCLC cells. [Figure 73]

[0188] 1 shows the efficacy of HC-5404 against A375 melanoma cells. [Figure 74]

[0189] 1 shows tumor growth curves for the MFE280 MSS endometrial cancer model treated with HC-5404 (30 mg / kg) administered orally twice daily, lenvatinib (10 mg / kg) administered orally once daily, or a combination thereof. [Figure 75]

[0190] 1 shows tumor growth curves of GA0114 PDX gastric cancer model treated with HC-5404 and DC-101. [Figure 76]

[0191] 1 shows tumor growth curves for the TM01125 PDX pancreatic cancer model treated with HC-5404 and sunitinib. [Figure 77]

[0192] 1 shows tumor growth curves for the MFE280 model, a human endometrial cancer model, treated with oral administration of HC-5404 twice daily, oral administration of lenvatinib (10 mg / kg) once daily, or a combination thereof. [Figure 78]

[0193] 1 shows the body weight curves of the MFE280 model, a human endometrial cancer model, treated with oral administration of HC-5404 twice daily, oral administration of lenvatinib (10 mg / kg) once daily, or a combination thereof. [Figure 79]

[0194] This study demonstrates the effect of HC-5404 in combination with lenvatinib on the MFE280 human endometrial cancer model. IHC images and quantification of FFPE sections from MFE280 tumors treated with HC-5404 (39 mg / kg; PO; BID), lenvatinib (10 mg / kg; PO; QD), or the combination for 7 days. Tumor samples were harvested 2 hours after the morning dose. [Figure 80]

[0195] This study demonstrates the effect of HC-5404 in combination with lenvatinib on the MFE280 human endometrial cancer model. IHC images and quantification of FFPE sections from MFE280 tumors treated with HC-5404 (39 mg / kg; PO; BID), lenvatinib (10 mg / kg; PO; QD), or the combination for 7 days. Tumor samples were collected 2 hours after the morning dose. [Figure 81]

[0196] We demonstrate the efficacy of HC-5404 in combination with DC-101 in a gastric PDX model. Simple Western protein analysis (JESS) of pPERK, PERK, and five proteins involved in amino acid metabolism: asparagine synthetase (ASNS), phosphoglycerate dehydrogenase (PHGDH), phosphoserine aminotransferase 1 (PSAT1), phosphoserine phosphatase (PSPH), and serine hydroxymethyltransferase 1 (SHMT1) was performed. The gastric cancer patient-derived xenograft (PDX) model GA0114-R19P6 was treated with HC-5404 (30 mg / kg; PO; BID), DC101 (20 mg / kg; IP; BIW), or their combination for 7 days, with samples collected 1 hour after the final morning dose. These proteins may serve as biomarkers of HC-5404 activity in vivo. [Figure 82]

[0197] Demonstrates the effect of HC-5404 in combination with DC-101 in a gastric PDX model. Quantification of Simple Western signals presented in panel Figure 81. [Figure 83]

[0198] Tumor growth curves of the CAPAN2 PDAC model treated with HC-5404 in combination with gemcitabine. [Figure 84]

[0199] Tumor growth curves in an MB49 bladder cancer model treated with HC-5404 at 3, 10, or 30 mg / kg PO BID for 14 days in combination with an anti-PD1 (IP; BIW) antibody. [Figure 85]

[0200] Tumor growth curves of the NCI-H660 NEPC model treated with HC-5404 in combination with docetaxel. [Figure 86]

[0201] Figure 1 shows plasma analysis from mice bearing 786-O tumors treated with 3, 10, or 30 mg / kg HC-5404 (PO; BID) as a single agent or in combination with cabozantinib (30 mg / kg; PO; QD). Plasma vascular endothelial growth factor A (VEGF-A) was dose-dependently reduced by HC-5404 as a single agent. [Figure 87]

[0202]

[0033] Figure 1 shows plasma analysis from mice bearing 786-O tumors treated with 3, 10, or 30 mg / kg HC-5404 (PO; BID) as a single agent or in combination with cabozantinib (30 mg / kg; PO; QD). In mouse plasma, hepatocyte growth factor (HGF) was reduced by the combination of HC-5404 and cabozantinib. [Figure 88]

[0203]

[0033] Figure 1 shows plasma analysis from mice bearing 786-O tumors treated with 3, 10, or 30 mg / kg HC-5404 (PO; BID) as a single agent or in combination with cabozantinib (30 mg / kg; PO; QD). Interleukin-8 (IL-8) was reduced in mouse plasma by the combination of HC-5404 and cabozantinib. [Figure 89]

[0204]

[0033] Figure 1 shows plasma analysis from mice bearing 786-O tumors treated with 3, 10, or 30 mg / kg HC-5404 (PO; BID) as a single agent or in combination with cabozantinib (30 mg / kg; PO; QD). Lymphatic endothelial hyaluronan receptor-1 (LYVE-1) was dose-dependently reduced by HC-5404, either as monotherapy or in combination with cabozantinib. [Figure 90]

[0205]

[0033] Figure 1 shows plasma analysis from mice bearing 786-O tumors treated with 3, 10, or 30 mg / kg HC-5404 (PO; BID) as a single agent or in combination with cabozantinib (30 mg / kg; PO; QD). Syndecans were reduced by HC-5404 as a single agent or in combination with cabozantinib. [Figure 91]

[0206] Tumor growth curves for mice bearing 786-O xenografts treated with axitinib (30 mg / kg; PO; BID) for 14 days and then re-randomized into four groups once tumors progressed (approximately doubled in size). 786-O xenografts were allowed to progress on axitinib for 2 weeks, after which HC-5404 was added to the regimen. Data presented are mean ± SEM. [Figure 92]

[0207] Mice bearing 786-O xenografts were treated with axitinib (30 mg / kg; PO; BID) for 14 days and randomized to tumor progression (approximately doubled in size). Quantification of IHC images from FFPE tumor sections sampled 7 days after treatment is shown for HC-5404-treated mice. Top left: Both HC-5404 and axitinib reduced the proportion of smooth muscle actin (SMA) as monotherapy, but the proportion was further reduced by combination treatment. Top right: The proportion of cells expressing Meca32 was reduced in the combination group compared to vehicle. Meca32 is a vascular endothelial cell marker. Bottom: The proportion of cells expressing the vascular marker CD31 was reduced by axitinib, but the effect was enhanced by the addition of HC-5404 in the combination group. [Figure 93]

[0208] Mice bearing 786-O xenografts were treated with axitinib (30 mg / kg; PO; BID) for 14 days and randomized when tumors progressed (approximately doubled in size). Quantification of IHC images from FFPE tumor sections sampled 7 days after treatment in mice treated with HC-5404 is shown. Tumors randomized to the combination group showed a decreased proportion of cells expressing the pericyte markers NG2 and MCAM. [Figure 94]

[0209] Tumor growth curves of the 786-O RCC tumor model treated with HC-5404 (30 mg / kg; PO; BID) and DC-101 (15 mg / kg; IP; BIW) for 28 days. Mean tumor volume ± SEM. [Figure 95]

[0210] Tumor growth curves of A498 RCC tumor models treated with HC-5404 (30 mg / kg; PO; BID) and DC-101 (15 mg / kg; IP; BIW) for 28 days. Mean tumor volume ± SEM. [Figure 96]

[0211] 1 shows tumor growth curves for the RENCA RCC tumor model treated with HC-5404 (30 mg / kg; PO; BID), axitinib (30 mg / kg; PO; BID), or their combination. [Figure 97]

[0212] Tumor growth curves of the RXF-393 RCC tumor model treated with HC-5404 (30 mg / kg; PO; BID), axitinib (30 mg / kg; PO; BID), or their combination. [Figure 98]

[0213] Tumor growth curves of the RXF-2282 RCC tumor model treated with HC-5404 (30 mg / kg; PO; BID), axitinib (30 mg / kg; PO; BID), or their combination. [Figure 99]

[0214] 10 shows tumor growth curves for the RXF-2178 RCC tumor model treated with HC-5404 (30 mg / kg; PO; BID), axitinib (30 mg / kg; PO; BID), or their combination. [Figure 100]

[0215] 16 shows tumor growth curves for the RXF-2502 RCC tumor model treated with HC-5404 (30 mg / kg; PO; BID), axitinib (30 mg / kg; PO; BID), or their combination. [Figure 101]

[0216] 1 shows tumor growth curves for SMTCA75 RCC tumor models treated with HC-5404 (30 mg / kg; PO; BID), axitinib (30 mg / kg; PO; BID), or their combination. [Figure 102]

[0217] 1 shows tumor growth curves for the RXF-616 RCC tumor model treated with HC-5404 (30 mg / kg; PO; BID), axitinib (30 mg / kg; PO; BID), or their combination. [Figure 103]

[0218] 1 shows tumor growth curves for the RXF-2304 RCC tumor model treated with HC-5404 (30 mg / kg; PO; BID), axitinib (30 mg / kg; PO; BID), or their combination. [Figure 104]

[0219] 1 shows tumor growth curves for the RXF-488 RCC tumor model treated with HC-5404 (30 mg / kg; PO; BID), axitinib (30 mg / kg; PO; BID), or their combination. [Figure 105]

[0220] 16 shows tumor growth curves for the RXF-2667 RCC tumor model treated with HC-5404 (30 mg / kg; PO; BID), axitinib (30 mg / kg; PO; BID), or their combination. [Figure 106]

[0221] 1 shows tumor growth curves for the RXF-1220 RCC tumor model treated with HC-5404 (30 mg / kg; PO; BID), axitinib (30 mg / kg; PO; BID), or their combination. [Figure 107]

[0222] Tumor growth curves of the RXF-2783 RCC tumor model treated with HC-5404 (30 mg / kg; PO; BID), axitinib (30 mg / kg; PO; BID), or their combination. [Figure 108]

[0223] 1 shows tumor growth curves for the RXF-631 RCC tumor model treated with HC-5404 (30 mg / kg; PO; BID), axitinib (30 mg / kg; PO; BID), or their combination. DETAILED DESCRIPTION OF THE INVENTION

[0090] Detailed Description

[0224] As generally described herein, the present disclosure provides methods of treating cancer in a subject in need of such treatment (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma). The present disclosure also provides methods of activating the immune system of a subject in need thereof (e.g., a subject with cancer). The methods described herein generally include administering to the subject a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapy (e.g., an anti-PD-1 antibody).

[0091]

[0225] The present disclosure also provides methods of treating cancer in a subject in need thereof (e.g., bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer), and thyroid cancer). The methods described herein generally include administering to a subject a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-angiogenic agent. Also disclosed herein are methods comprising administering to a subject a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, a therapeutically effective amount of an anti-angiogenic agent, and a therapeutically effective amount of an immunotherapy.

[0092] definition

[0226] To facilitate understanding of this invention, a number of terms and phrases are defined below.

[0093]

[0227] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Abbreviations used herein have their conventional meaning within the chemical and biological arts. The chemical structures and formulas set forth herein are prepared in accordance with the standard rules of chemical valency known in the chemical arts.

[0094]

[0228] Throughout this description, when compositions are described as having, containing, or comprising specific ingredients, or processes and methods are described as having, containing, or comprising specific steps, it is contemplated that there are compositions of the invention that consist essentially of or consist of the listed ingredients, and processes and methods of the invention that consist essentially of or consist of the listed processing steps.

[0095]

[0229] In this application, when an element or component is said to be included in and / or selected from a list of described elements or components, it is understood that the element or component can be any one of the described elements or components, or that the element or component can be selected from a group consisting of two or more of the described elements or components.

[0096]

[0230] Furthermore, it should be understood that elements and / or features of the compositions or methods described herein, whether expressly or implicitly stated herein, may be combined in various ways without departing from the spirit and scope of the invention. For example, where a particular compound is referenced, that compound may be used in various embodiments of the compositions and / or methods of the invention, unless otherwise understood from the context. In other words, within this application, embodiments are described and depicted in a manner that allows a clear and concise application to be described and depicted, but it is intended and will be understood that the embodiments may be combined in various ways or separated apart without departing from the present teachings and one or more inventions. For example, it will be understood that all of the features described and depicted herein may be applicable to all one or more aspects of the invention described and depicted herein.

[0097]

[0231] The articles "a" and "an" are used in this disclosure, unless the context is inappropriate, to refer to one or to more than one (i.e., to at least one) of the grammatical referent of the article. By way of example, "an element" means one element or more than one element.

[0098]

[0232] The term "and / or" is used in this disclosure to mean either "and" or "or," unless otherwise indicated.

[0099]

[0233] The phrase "at least one of" should be understood to include each of the listed objects following the phrase individually, as well as various combinations of two or more of the listed objects, unless the context and usage clearly indicates otherwise. The phrase "and / or" should be understood to have the same meaning when more than two listed objects are involved, unless the context clearly indicates otherwise.

[0100]

[0234] Use of the terms "include," "includes," "including," "have," "has," "having," "contain," "contains," or "containing," including grammatical equivalents thereof, should be understood to be generally open-ended and non-limiting, e.g., not excluding additional, unrecited elements or steps, unless otherwise specifically stated or understood from the context.

[0101]

[0235] Where the term "about" is used before a quantitative value, the invention also includes the specific quantitative value itself, unless otherwise specifically stated. As used herein, the term "about" refers to a ±10% variation from the nominal value, unless otherwise indicated or inferred from the context.

[0102]

[0236] At various places in the specification, variables or parameters are disclosed in group or range format, and the description is specifically intended to include every individual subcombination of the members of such groups and ranges. For example, an integer in the range of 0 to 40 is specifically intended to disclose 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, and 40 individually, and an integer in the range of 1 to 20 is specifically intended to disclose 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20 individually.

[0103]

[0237] Any examples or use of example language, such as "such as" or "including" herein, are intended merely to better illustrate the invention and do not impose limitations on the scope of the invention unless claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0104]

[0238] As a general matter, compositions with percentage designations are by weight unless otherwise specified. Furthermore, if a variable is not accompanied by a definition, the previous definition of that variable takes precedence.

[0105]

[0239] As used herein, a "pharmaceutical composition" or "pharmaceutical formulation" refers to a combination of an active agent with an inert or active carrier that makes the composition particularly suitable for diagnostic or therapeutic use in vivo or ex vivo.

[0106]

[0240] "Pharmaceutically acceptable" means approved or expected to be approved by a federal or state regulatory authority or a corresponding authority in a country other than the United States for use in animals, and more particularly in humans, or listed in the United States Pharmacopoeia or other generally recognized pharmacopeia.

[0107]

[0241] As used herein, "pharmaceutically acceptable salt" refers to any salt of an acidic or basic group that may be present in a compound of the present invention (e.g., a compound of Formula (I)), which is compatible with pharmaceutical administration.

[0108]

[0242] As known to those skilled in the art, "salts" of compounds can be derived from inorganic or organic acids and bases. Examples of acids include, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, p-toluenesulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, ethanesulfonic acid, formic acid, benzoic acid, malonic acid, naphthalene-2-sulfonic acid, and benzenesulfonic acid. Other acids, such as oxalic acid, while not themselves pharmaceutically acceptable, can be used to prepare salts useful as intermediates in obtaining the compounds described herein and their pharmaceutically acceptable acid addition salts.

[0109]

[0243] Examples of bases include, but are not limited to, alkali metal (e.g., sodium and potassium) hydroxides, alkaline earth metal (e.g., magnesium and calcium) hydroxides, ammonia, and bases of formula NW 4+ where W is C 1~4 alkyl).

[0110]

[0244] Examples of salts include, but are not limited to, acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, flucoheptanoate, glycerophosphate, hemifumarate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, undecanoate, and the like. Other examples of salts include Na + , K. + , Ca 2+ , N.H. 4+ and N.W. 4+ (Wherein W is C 1~4 These include anions of the compounds of the present invention that form compounds with suitable cations, such as cations that may be alkyl groups.

[0111]

[0245] For therapeutic use, salts of the compounds of the invention (e.g., compounds of Formula (I)) are contemplated as pharmaceutically acceptable. However, salts of acids and bases that are non-pharmaceutically acceptable may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound.

[0112]

[0246] As used herein, "pharmaceutically acceptable excipient" refers to a substance that facilitates administration of and / or absorption of an active agent by a subject and can be included in the compositions of the present invention without causing significant adverse toxicological effects to the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline, phosphate-buffered saline solution, emulsions (e.g., oil / water or water / oil emulsions), lactated Ringer's solution, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (such as Ringer's solution), alcohol, oils, gelatin, carbohydrates, fatty acid esters, and coloring agents. Such preparations can be sterilized and, if necessary, mixed with auxiliary substances that do not adversely react with the compounds of the present invention, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts that affect osmotic pressure, buffers, coloring agents, and / or aromatic substances. For examples of excipients, see Martin, Remington's Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, PA (1975).

[0113]

[0247] "Subjects" to which administration is contemplated include, but are not limited to, humans (i.e., male or female of any age, e.g., a pediatric subject (e.g., infant, child, adolescent) or an adult subject (e.g., young adult, middle-aged adult, or elderly adult)) and / or non-human animals, such as mammals, e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys), cows, pigs, horses, sheep, goats, rodents, cats, and / or dogs. In certain embodiments, the subject is a human. In certain embodiments, the subject is an adult human. In certain embodiments, the subject is a non-human animal.

[0114]

[0248] As used herein, "solid dosage form" means one or more pharmaceutical doses in solid form, such as tablets, capsules, granules, powders, sachets, reconstitutable powders, dry powder inhalants, and chewable tablets.

[0115]

[0249] As used herein, "administering" refers to oral administration, administration as a suppository, topical contact, intravenous administration, parenteral administration, intraperitoneal administration, intramuscular administration, intralesional administration, intrathecal administration, intracranial administration, intranasal administration, or subcutaneous administration to a subject, or implantation of a sustained-release device, such as a mini-osmotic pump. Administration is by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, for example, intravenous, intramuscular, intraarterial, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, transdermal patches, and the like. "Co-administering" means that a composition described herein is administered at the same time as, immediately before, or immediately after the administration of one or more additional therapies (e.g., anti-cancer agents, chemotherapeutics, or immunotherapies). The compound of formula (I) or its pharmaceutically acceptable salt can be administered to a patient alone or co-administered. Co-administration is meant to include simultaneous or sequential administration of the compounds individually or in combination (two or more compounds or drugs). Thus, the preparation can also be combined with other active substances, if necessary (e.g., to reduce metabolic degradation).

[0116]

[0250] As used herein, "fasted state" means at least 1 hour before or at least 2 hours after a subject ingests food.

[0117]

[0251] The terms "disease," "disorder," and "condition" are used interchangeably herein.

[0118]

[0252] As used herein and unless otherwise specified, the terms "treat," "treating," and "treatment" contemplate actions taken while a subject is afflicted with a specified disease, disorder, or condition, whereby the severity of the disease, disorder, or condition is reduced or the progression of the disease, disorder, or condition is delayed or slowed (e.g., "therapeutic treatment").

[0119]

[0253] In general, an "effective amount" or "therapeutically effective amount" of a compound (e.g., a compound of Formula (I) or a pharmaceutically acceptable salt thereof) refers to an amount sufficient to elicit a desired biological response in a subject in need thereof, such as an amount sufficient to treat cancer or activate the immune system. As one of ordinary skill in the art would understand, the therapeutically effective amount of a compound of the present disclosure may vary depending on factors such as the desired biological endpoint, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the age, weight, health, and condition of the subject.

[0120]

[0254] As used herein, the term "activating" refers to the stimulation of immune cells (e.g., T cells, NK cells, B cells, dendritic cells, or macrophages) resulting in cell proliferation, maturation, cytokine production, and / or induction of regulatory or effector functions. Methods for assessing immune cell activation or function are well known in the art and include, but are not limited to, proliferation assays such as BRDU and thymidine incorporation, cytotoxicity assays such as chromium release assays, cytokine secretion assays such as intracellular cytokine staining, ELISPOT, and ELISA, and expression assays of activation markers such as CD25 and CD69 using flow cytometry and multimer (e.g., tetramer) assays.

[0121]

[0255] As used herein, the term "myeloid-derived suppressor cells" or "MDSC" refers to a group of immature myeloid cells that regulate the activity of various effector cells and antigen-presenting cells, including, inter alia, T cells, NK cells, dendritic cells, and macrophages. Myeloid-derived suppressor cells are distinguished by their gene expression profile, which includes B7-1 (CD80), B7-H1 (PD-L1), CCR2, CD1d, CD1d1, CD2, CD31 (PECAM-1), CD43, CD44, complement components C5aR1, F4 / 80 (EMR1), Fcy RIII (CD16), Fcy RII (CD32), Fcy RIIA (CD32a), Fcy RIIB (CD32b), Fcy RIIB / C (CD32b / c), Fcy RIIC (CD32c), Fcy RIIIA (CD16A), Fcy RIIIB (CD16b), galectin-3, GP130, Gr-1 (Ly-6G), ICAM-1 (CD54), IL-1RI, IL-4Rα, IL-6Rα, integrin αL (CD49d), integrin αL (CD11a), integrin αM (CD11b), M-CSFR, MGL1 (CD301a), MGL1 / 2 (CD301a / b), MGL2 (CD301b), nitric oxide, PSGL-1 (CD162), L-selectin (CD62L), Siglec-3 (CD33), transferrin receptor (TfR), VEGFR1 (Flt-1), and VEGFR2 (KDR or Flk-1). In particular, MDSCs do not express proteins selected from the group consisting of B7-2 (CD86), B7-H4, CD11c, CD14, CD21, CD23 (FcεRII), CD34, CD35, CD40 (TNFRSF5), CD117 (c-kit), HLA-DR, and Sca-1 (Ly6).

[0122]

[0256] As used herein, the phrase "tumor-infiltrating lymphocytes" refers to white blood cells (e.g., T cells and / or NK cells) of a subject with cancer (e.g., a cancer described herein) that are resident in the tumor or that have otherwise left the circulation (blood or lymph) and migrated to the tumor.

[0123]

[0257] As used herein, the term "immunotherapy" refers to the treatment of disease by inducing, enhancing, or suppressing an immune response. Immunotherapies designed to elicit or amplify an immune response are classified as activating immunotherapies, while immunotherapies that reduce or suppress an immune response are classified as suppressing immunotherapies.

[0124]

[0258] As used herein, the phrase "immune checkpoint inhibitor" refers to drugs that block specific proteins made by some types of immune system cells, such as T cells, and by some cancer cells. These proteins help suppress the immune response and can prevent T cells from killing cancer cells. Blocking these proteins restores immune system function and unleashes the immune system, allowing T cells to kill cancer cells.

[0125]

[0259] As used herein, the term "anti-angiogenic agent" refers to an agent that modulates (e.g., inhibits) angiogenesis, vasculogenesis, or vascular permeability. Anti-angiogenic agents include agents that block the angiogenic activity of angiogenic factors or their receptors.

[0126]

[0260] As used herein, the term "antibody" refers to a functional component of serum and is often referred to either as a group of molecules (antibodies or immunoglobulins) or as a single molecule (the antibody molecule or immunoglobulin molecule). Antibodies have the ability to bind to or react with a specific antigenic determinant (the antigen or antigenic epitope), which can lead to the induction of immune effector mechanisms. Individual antibodies are usually considered monospecific, and antibody compositions can be monoclonal (i.e., consisting of identical antibody molecules) or polyclonal (i.e., consisting of two or more different antibodies reacting with the same or different epitopes on the same antigen or on distinct different antigens). Each antibody has a unique structure that enables specific binding to its corresponding antigen, and all naturally occurring antibodies have the same overall basic structure of two identical light chains and two identical heavy chains. Antibodies are also collectively known as immunoglobulins. The antibody may be of human or non-human (e.g., rodent such as mouse, canine, camelid, etc.) origin (e.g., may have a sequence that originally occurred in a human or non-human cell or organism), or may be or include, for example, a chimeric, humanized, reshaped, or reformatted antibody based on (or in some embodiments based on an antigen-binding portion of) such a human or non-human antibody.

[0127]

[0261] As used herein, "biosimilar" is used to describe a biological agent that is highly similar to a reference biological agent. The term is generally used to describe a follow-on version of a biological agent (e.g., from a different source) that is not expected to be exactly the same as the reference biological agent.

[0128]

[0262] As used herein, the term "antagonist" refers to an agent that (i) reduces or inhibits one or more effects of another agent; and / or (ii) reduces or inhibits one or more biological events. In some embodiments, an antagonist may reduce the level and / or activity of one or more agents that it targets. In some embodiments, an antagonist may be a receptor antagonist, e.g., a receptor ligand or drug, that blocks or attenuates a biological response by binding to and blocking the receptor rather than activating the receptor as an agonist does.

[0129]

[0263] As used herein, the term "agonist" refers to an agent that (i) increases or induces one or more effects of another agent; and / or (ii) increases or induces one or more biological events. In some embodiments, an agonist may increase the level and / or activity of one or more agents that it targets.

[0130] PERK inhibitors

[0264] The compound of formula (I) as depicted below is a selective PERK inhibitor and is (R)-2-amino-5-(4-(2-(3,5-difluorophenyl)-2-hydroxyacetamido)-2-methylphenyl)-N-isopropylnicotinamide: [ka] It is also known as

[0131]

[0265] The compound of formula (I) may also be referred to throughout this disclosure as HC-5404. The hemifumarate salt of the compound of formula (I) may also be referred to throughout this disclosure as HC-5404-FU. Methods for chemically synthesizing the compound of formula (I) and its hemifumarate salt are described in Example 1.

[0132]

[0266] In one aspect, provided herein are methods for treating cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, malignant melanoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment, comprising administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor).

[0133]

[0267] In another aspect, provided herein are methods for administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor) for the treatment of cancer (e.g., a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0134]

[0268] In another aspect, provided herein are methods of administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody for the treatment of cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, malignant melanoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0135]

[0269] In another aspect, provided herein are methods of administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody for the treatment of cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0136]

[0270] In another aspect, provided herein is a method of administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor) to activate the immune system of a subject in need thereof (e.g., a subject having a cancer described herein).

[0137]

[0271] In another aspect, provided herein is a method of administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody to activate the immune system of a subject in need thereof (e.g., a subject having a cancer described herein).

[0138]

[0272] In various embodiments, provided herein are methods for treating cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, malignant melanoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment, comprising administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor).

[0139]

[0273] In various embodiments, provided herein are methods for treating cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment, comprising administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor).

[0140]

[0274] In various embodiments, provided herein are methods for treating cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, malignant melanoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment, comprising administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) and a therapeutically effective amount of an anti-PD-1 antibody.

[0141]

[0275] In various embodiments, provided herein are methods for treating cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment, comprising administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) and a therapeutically effective amount of an anti-PD-1 antibody.

[0142]

[0276] In various embodiments, provided herein are methods for administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor) to activate the immune system of a subject in need thereof (e.g., a subject having a cancer described herein).

[0143]

[0277] In various embodiments, provided herein are methods for administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) and a therapeutically effective amount of an anti-PD-1 antibody to activate the immune system of a subject in need thereof (e.g., a subject having a cancer described herein).

[0144]

[0278] In certain embodiments, the immunotherapy is a PD-1 or PD-L1 inhibitor, hi certain embodiments, the immunotherapy is an anti-PD-1 antibody.

[0145]

[0279] In another aspect, provided herein is a method of administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-angiogenic agent (e.g., a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI)) for the treatment of cancer in a subject in need thereof (e.g., in some embodiments, a solid tumor selected from the group consisting of bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer (e.g., malignant melanoma)), and thyroid cancer).

[0146]

[0280] In another aspect, provided herein is a method of administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-angiogenic agent (e.g., a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI)) for the treatment of cancer in a subject in need thereof (e.g., in some embodiments, a solid tumor selected from the group consisting of bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer), and thyroid cancer).

[0147]

[0281] In another aspect, the present disclosure relates to cancer (e.g., in some embodiments, bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer (NSCLC)), or tumors of the genotype (e.g., urinary tract cancer ...

[0013] Provided is a method for treating cancer in a subject in need of such treatment (a solid tumor selected from the group consisting of: ovarian cancer, pancreatic cancer, prostate cancer, skin cancer (e.g., malignant melanoma), and thyroid cancer), comprising administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-angiogenic agent (e.g., a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI)), wherein the subject is being treated with an anti-cancer therapy.

[0148]

[0282] In another aspect, provided herein is a method of administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-angiogenic agent (e.g., a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI)) for the treatment of cancer in a subject in need thereof (e.g., in some embodiments, a solid tumor selected from the group consisting of bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer), and thyroid cancer), wherein the subject is being treated with an anti-cancer therapy.

[0149]

[0283] In another aspect, provided herein is a method of administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof for the treatment of cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer (e.g., malignant melanoma)), and thyroid cancer) in a subject receiving a therapeutically effective amount of anti-angiogenic agent therapy.

[0150]

[0284] In another aspect, provided herein is a method of administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof for the treatment of cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer), and thyroid cancer) in a subject receiving a therapeutically effective amount of anti-angiogenic agent therapy.

[0151]

[0285] In various embodiments, the methods provided herein further comprise administering to the subject a therapeutically effective amount of an immunotherapy (e.g., an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody).

[0152]

[0286] In one aspect, the present disclosure relates to cancer (e.g., in some embodiments, bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, pre-cancerous carcinoma, ovarian cancer, pancreatic cancer ... Provided is a method for treating cancer in a subject in need of such treatment (a solid tumor selected from the group consisting of prostate cancer, skin cancer (e.g., malignant melanoma) and thyroid cancer), comprising administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, a therapeutically effective amount of an anti-angiogenic agent (e.g., a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI)), and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor, such as an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody).

[0153]

[0287] In one aspect, the present disclosure relates to cancer (e.g., in some embodiments, bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, or ovarian cancer). and a solid tumor selected from the group consisting of thyroid cancer, pancreatic cancer, prostate cancer, and skin cancer), and thyroid cancer, in a subject in need of such treatment. The method comprises administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, a therapeutically effective amount of an anti-angiogenic agent (e.g., a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI)), and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor, such as an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody).

[0154]

[0288] In another aspect, the present disclosure relates to cancer (e.g., in some embodiments, bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer (e.g., Provided is a method for treating cancer in a subject in need of such treatment (a solid tumor selected from the group consisting of thyroid cancer, malignant melanoma, and thyroid cancer), comprising administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, a therapeutically effective amount of an anti-angiogenic agent (e.g., a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI)), and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor, such as an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody), wherein the subject is being treated with an anti-cancer therapy.

[0155]

[0289] In another aspect, the present disclosure relates to cancer (e.g., in some embodiments, bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer, and the like). and thyroid cancer), the subject is being treated with an anti-cancer therapy.

[0156]

[0290] In another aspect, provided herein is a method for treating a cancer (e.g., in some embodiments, bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as clear cell renal carcinoma (ccRCC)) in a subject receiving a therapeutically effective amount of anti-angiogenic agent therapy. and a solid tumor selected from the group consisting of ovarian cancer, pancreatic cancer, prostate cancer, skin cancer (e.g., malignant melanoma), and thyroid cancer, comprising administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor, such as an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody) for the treatment of a tumor of the thyroid gland (e.g., a thyroid cancer), a thyroid cancer ...

[0157]

[0291] In another aspect, provided herein are methods of administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor, e.g., an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody) for the treatment of cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer), and thyroid cancer) in a subject receiving a therapeutically effective amount of anti-angiogenic agent therapy.

[0158]

[0292] In certain embodiments, a pharmaceutically acceptable salt of a compound of formula (I) is a fumarate salt. In certain embodiments, a pharmaceutically acceptable salt of a compound of formula (I) is a hemifumarate salt.

[0159] Antiangiogenic agents

[0293] Antiangiogenic agents are limited by primary and secondary resistance mechanisms, including VEGFR-TKI induction of hypoxia and nutrient deprivation, which drive ER stress. Tumors circumvent harmful ER stress by activating the PERK branch of the integrated stress response, which halts translation globally and restores proteostasis.

[0160]

[0294] In some embodiments, the antiangiogenic agent is an antibody or a biosimilar thereof. In some embodiments, the antiangiogenic agent is a small molecule or a pharmaceutically acceptable salt thereof. In some embodiments, the antiangiogenic agent is a vascular endothelial growth factor receptor (VEGFR) modulator. In some embodiments, the antiangiogenic agent is a vascular endothelial growth factor (VEGF)-targeting antibody. In some embodiments, the antiangiogenic agent is a tyrosine kinase inhibitor (TKI). In some embodiments, the antiangiogenic agent is a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI).

[0161]

[0295] Examples of anti-angiogenic agents include, but are not limited to, apatinib, axitinib, brivanib, cabozantinib, canertinib, cediranib, dasatinib, erlotinib, gefitinib, leflunomide, 42envatinib, motesanib, nilotinib, nintedanib, pazopanib, regorafenib, semaxinib, sorafenib, sunitinib, tivozanib, vandetanib, vatalanib, XL0192, bevacizumab, ramucirumab, and pharmaceutically acceptable salts or biosimilars thereof.

[0162] Pharmaceutical Composition

[0296] Provided herein are pharmaceutical compositions generally comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.

[0163]

[0297] In one aspect, provided herein is a method of administering a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, one or more pharmaceutically acceptable excipients, and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor) for the treatment of cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, malignant melanoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0164]

[0298] In another aspect, provided herein is a method of administering a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, one or more pharmaceutically acceptable excipients, and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor) for the treatment of cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0165]

[0299] In another aspect, provided herein is a method of administering a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, one or more pharmaceutically acceptable excipients, and a therapeutically effective amount of an anti-PD-1 antibody for the treatment of cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, malignant melanoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0166]

[0300] In another aspect, provided herein is a method of administering a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, one or more pharmaceutically acceptable excipients, and a therapeutically effective amount of an anti-PD-1 antibody for the treatment of cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0167]

[0301] In another aspect, provided herein is a method of administering a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, one or more pharmaceutically acceptable excipients, and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor) to activate the immune system of a subject in need thereof (e.g., a subject having a cancer described herein).

[0168]

[0302] In another aspect, provided herein is a method of administering a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, one or more pharmaceutically acceptable excipients, and a therapeutically effective amount of an anti-PD-1 antibody, to activate the immune system of a subject in need thereof (e.g., a subject having a cancer described herein).

[0169]

[0303] In another aspect, provided herein is a method of administering a pharmaceutical composition comprising a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I), one or more pharmaceutically acceptable excipients, and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor) for the treatment of cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, malignant melanoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, and urothelial carcinoma) in a subject in need of such treatment.

[0170]

[0304] In another aspect, provided herein are methods of administering a pharmaceutical composition comprising a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I), one or more pharmaceutically acceptable excipients, and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor) for the treatment of cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, and urothelial carcinoma) in a subject in need of such treatment.

[0171]

[0305] In another aspect, provided herein are methods of administering a pharmaceutical composition comprising a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I), one or more pharmaceutically acceptable excipients, and a therapeutically effective amount of an anti-PD-1 antibody for the treatment of cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, malignant melanoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0172]

[0306] In another aspect, provided herein are methods of administering a pharmaceutical composition comprising a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I), one or more pharmaceutically acceptable excipients, and a therapeutically effective amount of an anti-PD-1 antibody for the treatment of cancer (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0173]

[0307] In another aspect, provided herein is a method of administering a therapeutically effective amount of a pharmaceutical composition comprising a pharmaceutically acceptable salt of a compound of Formula (I), one or more pharmaceutically acceptable excipients, and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor) to activate the immune system of a subject in need thereof (e.g., a subject having a cancer described herein).

[0174]

[0308] In another aspect, provided herein is a method of administering a pharmaceutical composition comprising a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I), one or more pharmaceutically acceptable excipients, and a therapeutically effective amount of an anti-PD-1 antibody, to activate the immune system of a subject in need thereof (e.g., a subject having a cancer described herein).

[0175]

[0309] In another aspect, provided herein is a pharmaceutical composition for the treatment of cancer (e.g., a cancer described herein), comprising a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, to be administered in conjunction with a therapeutically effective amount of immunotherapy (e.g., an immune checkpoint inhibitor).

[0176]

[0310] In another aspect, provided herein is a pharmaceutical composition for the treatment of cancer (e.g., a cancer described herein), comprising a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, to be administered in combination with a therapeutically effective amount of an anti-PD-1 antibody.

[0177]

[0311] In certain embodiments, the immunotherapy is a PD-1 or PD-L1 inhibitor, hi certain embodiments, the immunotherapy is an anti-PD-1 antibody.

[0178]

[0312] In various embodiments, provided herein are pharmaceutical compositions comprising a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.

[0179]

[0313] In various embodiments, provided herein are pharmaceutical compositions comprising a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) and one or more pharmaceutically acceptable excipients.

[0180]

[0314] In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is from about 22 mg to about 226 mg, from about 35 mg to about 226 mg, from about 45 mg to about 226 mg, from about 55 mg to about 226 mg, from about 65 mg to about 226 mg, from about 75 mg to about 226 mg, from about 85 mg to about 226 mg, from about 95 mg to about 226 mg, from about 105 mg to about 226 mg, from about 115 mg to about 226 mg, from about 125 mg to about 226 mg, or from about 140 mg to about 226 mg. 200mg, about 135mg to about 226mg, about 145mg to about 226mg, about 155mg to about 226mg, about 165mg to about 226mg, about 175mg to about 226mg, about 185mg to about 226mg, about 195mg to about 226mg, about 2 2mg to about 190mg, about 22mg to about 180mg, about 22mg to about 170mg, about 22mg to about 160mg, about 22mg to about 150mg, about 22mg to about 140mg, about 22mg to about 130mg, about 22mg to about 120mg, about 22mg about 22 mg to about 110 mg, about 22 mg to about 100 mg, about 22 mg to about 90 mg, about 22 mg to about 80 mg, about 22 mg to about 70 mg, about 22 mg to about 60 mg, about 22 mg to about 50 mg, about 22 mg to about 40 mg, about 22 mg to about 30 mg, about 35 mg to about 185 mg, about 45 mg to about 175 mg, about 55 mg to about 165 mg, about 65 mg to about 155 mg, about 75 mg to about 145 mg, about 85 mg to about 135 mg, about 95 mg to about 125 mg, or about 105 mg to about 115 mg.

[0181]

[0315] In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is from about 177 mg to about 900 mg, from about 250 mg to about 900 mg, from about 300 mg to about 900 mg, from about 350 mg to about 900 mg, from about 400 mg to about 900 mg, from about 450 mg to about 900 mg, from about 500 mg to about 900 mg, from about 550 mg to about 900 mg, from about 600 mg to about 900 mg, from about 650 mg to about 900 mg, from about 700 mg to about 900 mg, from about 750 mg to about 900 mg, from about 800 mg to about 900 mg, from about 850 mg to about 900 mg, from about 177 mg to about 85 ... 7mg to about 800mg, about 177mg to about 750mg, about 177mg to about 700mg, about 177mg to about 650mg, about 177mg to about 600mg, about 177mg to about 550mg, about 177mg to about 500mg, about 177mg to about 450mg, about 177mg to about 400mg, about 177mg to about 350mg, about 177mg to about 300mg, about 177mg to about 250mg, about 250mg to about 850mg, about 300mg to about 800mg, about 350mg to about 750mg, about 400mg to about 700mg, about 450mg to about 650mg, about 500mg to about 600mg, or about 550mg to about 600mg.

[0182]

[0316] In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is from about 22 mg to about 451 mg, from about 50 mg to about 451 mg, from about 75 mg to about 451 mg, from about 100 mg to about 451 mg, from about 125 mg to about 451 mg, from about 150 mg to about 451 mg, from about 175 mg to about 451 mg, from about 200 mg to about 451 mg, or from about 22 mg to about 451 mg. mg, about 225 mg to about 451 mg, about 250 mg to about 451 mg, about 275 mg to about 451 mg, about 300 mg to about 451 mg, about 325 mg to about 451 mg, about 350 mg ~451mg, 375mg~451mg, 22mg~375mg, 22mg~350mg, 22mg~325mg, 22mg~300mg, 22mg~ Approximately 275 mg, approximately 22 mg to approximately 250 mg, approximately 22 mg to approximately 225 mg, approximately 22 mg to approximately 200 mg, approximately 22 mg to approximately 175 mg, approximately 22 mg to approximately 150 mg, approximately 22 mg to approximately 125mg, about 22mg to about 100mg, about 22mg to about 75mg, about 22mg to about 50mg, about 50mg to about 375mg, about 75mg to about 350mg, about 100mg to about 325 mg, about 125 mg to about 300 mg, about 150 mg to about 275 mg, about 175 mg to about 250 mg, about 200 mg to about 225 mg, about 50 mg to about 100 mg, about 100 mg to about 150 mg, about 150 mg to about 200 mg, about 200 mg to about 250 mg, about 250 mg to about 300 mg, about 300 mg to about 350 mg, or about 350 mg to about 451 mg.

[0183]

[0317] In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 22 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, about 115 mg, about 120 mg, about 125 mg, about 130 mg, about 135 mg, about 140 mg, about 145 mg, about 150 mg, about 155 mg, about 160 mg, about 165 mg, about 170 mg, about 175 mg, about 180 mg, about 185 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 400 mg, about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, about 500 mg, about 510 mg, about 520 mg, about 530 mg, about 540 mg, about mg, about 140mg, about 145mg, about 150mg, about 155mg, about 160mg, about 165mg, about 170mg, about 175mg, about 180mg, about 185mg, about 190mg, about 195mg, about 200mg, about 210 mg, about 220mg, about 230mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg, about 300mg, about 310mg, about 320mg, about 330mg, about 340mg, about 350 mg, about 360mg, about 370mg, about 380mg, about 390mg, about 400mg, about 410mg, about 420mg, about 430mg, about 440mg, about 450mg, about 460mg, about 470mg, about 480mg, about 490 mg, about 500mg, about 510mg, about 520mg, about 530mg, about 540mg, about 550mg, about 560mg, about 570mg, about 580mg, about 590mg, about 600mg, about 610mg, about 620mg, about 630 mg, about 640 mg, about 650 mg, about 660 mg, about 670 mg, about 680 mg, about 690 mg, about 700 mg, about 710 mg, about 720 mg, about 730 mg, about 740 mg, about 750 mg, about 760 mg, about 770 mg, about 780 mg, about 790 mg, about 800 mg, about 810 mg, about 820 mg, about 830 mg, about 840 mg, about 850 mg, about 860 mg, about 870 mg, about 880 mg, about 890 mg or about 900 mg.

[0184]

[0318] In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 22 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 50 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 75 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 100 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 125 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 150 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 175 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 200 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 250 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 300 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 350 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 400 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 450 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 500 mg. In certain embodiments, the amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 550 mg.In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 600 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 650 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 700 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 750 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 800 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 850 mg. In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is about 900 mg.

[0185]

[0319] In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is from about 22 mg to about 226 mg, from about 35 mg to about 226 mg, from about 45 mg to about 226 mg, from about 55 mg to about 226 mg, from about 65 mg to about 226 mg, from about 75 mg to about 226 mg, from about 85 mg to about 226 mg, from about 95 mg to about 226 mg, from about 105 mg to about 226 mg, from about 115 mg to about 226 mg, from about 125 mg to about 226 mg, from about 135 mg to about 226 mg, from about 145 mg to about 226 mg, from about 155 mg to about 226 mg, from about 165 mg to about 226 mg, from about 175 mg to about 226 mg, from about 185 mg to about 226 mg, from about 195 mg to about 226 mg, from about 22 mg to about 190 mg, or from about 22mg to about 180mg, about 22mg to about 170mg, about 22mg to about 160mg, about 22mg to about 150mg, about 22mg to about 140mg, about 22mg to about 130mg, Approximately 22mg to approximately 120mg, approximately 22mg to approximately 110mg, approximately 22mg to approximately 100mg, approximately 22mg to approximately 90mg, approximately 22mg to approximately 80mg, approximately 22mg to approximately 70mg, approximately 2 2 mg to about 60 mg, about 22 mg to about 50 mg, about 22 mg to about 40 mg, about 22 mg to about 30 mg, about 35 mg to about 185 mg, about 45 mg to about 175 mg, about 55 mg to about 165 mg, about 65 mg to about 155 mg, about 75 mg to about 145 mg, about 85 mg to about 135 mg, about 95 mg to about 125 mg, or about 105 mg to about 115 mg.

[0186]

[0320] In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is from about 177 mg to about 900 mg, from about 250 mg to about 900 mg, from about 300 mg to about 900 mg, from about 350 mg to about 900 mg, from about 400 mg to about 900 mg, from about 450 mg to about 900 mg, from about 500 mg to about 900 mg, from about 550 mg to about 900 mg, from about 600 mg to about 900 mg, from about 650 mg to about 900 mg, from about 700 mg to about 900 mg, from about 750 mg to about 900 mg, from about 800 mg to about 900 mg, from about 850 mg to about 900 mg, from about 177 mg to about 85 ... g to about 800 mg, about 177 mg to about 750 mg, about 177 mg to about 700 mg, about 177 mg to about 650 mg, about 177 mg to about 600 mg, about 177 mg to about 550 mg, about 177 mg to about 500 mg, about 177 mg to about 450 mg, about 177 mg to about 400 mg, about 177 mg to about 350 mg, about 177 mg to about 300 mg, about 177 mg to about 250 mg, about 250 mg to about 850 mg, about 300 mg to about 800 mg, about 350 mg to about 750 mg, about 400 mg to about 700 mg, about 450 mg to about 650 mg, about 500 mg to about 600 mg, or about 550 mg to about 600 mg.

[0187]

[0321] In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is from about 22 mg to about 451 mg, from about 50 mg to about 451 mg, from about 75 mg to about 451 mg, from about 100 mg to about 451 mg, from about 125 mg to about 451 mg, from about 150 mg to about 451 mg, from about 175 mg to about 451 mg, or from about 200 mg to about 451 mg. , about 225 mg to about 451 mg, about 250 mg to about 451 mg, about 275 mg to about 451 mg, about 300 mg to about 451 mg, about 325 mg to about 451 mg, about 350 mg to about 451mg, about 375mg to about 451mg, about 22mg to about 375mg, about 22mg to about 350mg, about 22mg to about 325mg, about 22mg to about 300mg, about 22mg to about 2 75mg, about 22mg to about 250mg, about 22mg to about 225mg, about 22mg to about 200mg, about 22mg to about 175mg, about 22mg to about 150mg, about 22mg to about 12 5mg, about 22mg to about 100mg, about 22mg to about 75mg, about 22mg to about 50mg, about 50mg to about 375mg, about 75mg to about 350mg, about 100mg to about 325m g, about 125 mg to about 300 mg, about 150 mg to about 275 mg, about 175 mg to about 250 mg, about 200 mg to about 225 mg, about 50 mg to about 100 mg, about 100 mg to about 150 mg, about 150 mg to about 200 mg, about 200 mg to about 250 mg, about 250 mg to about 300 mg, about 300 mg to about 350 mg, or about 350 mg to about 451 mg.

[0188]

[0322] In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 22 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, about 115 mg, about 120 mg, about 125 mg, about 130 mg, about 135 mg , about 140mg, about 145mg, about 150mg, about 155mg, about 160mg, about 165mg, about 170mg, about 175mg, about 180mg, about 185mg, about 190mg, about 195mg, about 200mg, about 210m g, about 220mg, about 230mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg, about 300mg, about 310mg, about 320mg, about 330mg, about 340mg, about 350m g, about 360mg, about 370mg, about 380mg, about 390mg, about 400mg, about 410mg, about 420mg, about 430mg, about 440mg, about 450mg, about 460mg, about 470mg, about 480mg, about 490 mg, about 500mg, about 510mg, about 520mg, about 530mg, about 540mg, about 550mg, about 560mg, about 570mg, about 580mg, about 590mg, about 600mg, about 610mg, about 620mg, about 630 mg, about 640 mg, about 650 mg, about 660 mg, about 670 mg, about 680 mg, about 690 mg, about 700 mg, about 710 mg, about 720 mg, about 730 mg, about 740 mg, about 750 mg, about 760 mg, about 770 mg, about 780 mg, about 790 mg, about 800 mg, about 810 mg, about 820 mg, about 830 mg, about 840 mg, about 850 mg, about 860 mg, about 870 mg, about 880 mg, about 890 mg or about 900 mg.

[0189]

[0323] In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 22 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 50 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 75 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 100 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 125 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 150 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 175 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 200 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 250 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 300 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 350 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 400 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 450 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 500 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 550 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 600 mg.In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 650 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 700 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 750 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 800 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 850 mg. In certain embodiments, the amount of the pharmaceutically acceptable salt of the compound of Formula (I) in the pharmaceutical compositions described herein is about 900 mg.

[0190]

[0324] In various embodiments, as used herein, (i) about 22 mg to about 451 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof; (ii) one or more pharmaceutically acceptable excipients; and A pharmaceutical composition comprising:

[0191]

[0325] In various embodiments, as used herein, (i) about 177 mg to about 900 mg of a pharmaceutically acceptable salt of a compound of Formula (I); (ii) one or more pharmaceutically acceptable excipients; and A pharmaceutical composition comprising:

[0192]

[0326] In another aspect, provided herein is a pharmaceutical composition comprising about 22 mg to about 177 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients, for the treatment of a cancer described herein (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, malignant melanoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0193]

[0327] In another aspect, provided herein is a pharmaceutical composition comprising about 22 mg to about 177 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients, for the treatment of a cancer described herein (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0194]

[0328] In another aspect, provided herein is a pharmaceutical composition comprising about 177 mg to about 900 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients, for the treatment of a cancer described herein (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, malignant melanoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0195]

[0329] In another aspect, provided herein is a pharmaceutical composition comprising about 177 mg to about 900 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients, for the treatment of a cancer described herein (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0196]

[0330] In another aspect, provided herein is a pharmaceutical composition comprising about 22 mg to about 226 mg of a pharmaceutically acceptable salt of the compound of Formula (I) and one or more pharmaceutically acceptable excipients for the treatment of a cancer described herein (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, malignant melanoma, M-cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0197]

[0331] In another aspect, provided herein is a pharmaceutical composition comprising about 22 mg to about 226 mg of a pharmaceutically acceptable salt of the compound of Formula (I) and one or more pharmaceutically acceptable excipients for the treatment of a cancer described herein (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, M-cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0198]

[0332] In another aspect, provided herein is a pharmaceutical composition comprising about 177 mg to about 900 mg of a pharmaceutically acceptable salt of the compound of Formula (I) and one or more pharmaceutically acceptable excipients for the treatment of a cancer described herein (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, malignant melanoma, M-cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0199]

[0333] In another aspect, provided herein is a pharmaceutical composition comprising about 177 mg to about 900 mg of a pharmaceutically acceptable salt of the compound of Formula (I) and one or more pharmaceutically acceptable excipients for the treatment of a cancer described herein (e.g., in some embodiments, a solid tumor selected from the group consisting of head and neck squamous cell carcinoma, M-cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma) in a subject in need of such treatment.

[0200]

[0334] In another aspect, provided herein is a pharmaceutical composition comprising about 22 mg to about 226 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients, for activating the immune system of a subject in need thereof (e.g., a subject having a cancer described herein).

[0201]

[0335] In another aspect, provided herein is a pharmaceutical composition comprising about 177 mg to about 900 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients, for activating the immune system of a subject in need thereof (e.g., a subject having a cancer described herein).

[0202]

[0336] In another aspect, provided herein is a pharmaceutical composition comprising about 22 mg to about 226 mg of a pharmaceutically acceptable salt of a compound of Formula (I) and one or more pharmaceutically acceptable excipients, for activating the immune system of a subject in need thereof (e.g., a subject having a cancer described herein).

[0203]

[0337] In another aspect, provided herein is a pharmaceutical composition comprising about 177 mg to about 900 mg of a pharmaceutically acceptable salt of a compound of Formula (I) and one or more pharmaceutically acceptable excipients, for activating the immune system of a subject in need thereof (e.g., a subject having a cancer described herein).

[0204]

[0338] In various embodiments, provided herein are pharmaceutical compositions comprising a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I), a therapeutically effective amount of an anti-angiogenic agent, and one or more pharmaceutically acceptable excipients.

[0205]

[0339] In certain embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof in the pharmaceutical compositions described herein is from about 22 mg to about 798 mg, from about 35 mg to about 798 mg, from about 44 mg to about 798 mg, from about 66 mg to about 798 mg, from about 88 mg to about 798 mg, from about 110 mg to about 798 mg, from about 113 mg to about 798 mg, from about 155 mg to about 798 mg, from about 177 mg to about 798 mg, from about 199 mg to about 798 mg, from about 221 mg to about 798 mg, from about 244 mg to about 798 mg, from about 266 mg to about 798 mg, from about 310 mg to about 798 mg, from about 355 mg to about 798 mg, from about 399 mg to about 798 mg, from about 443 mg to about 798 mg, mg, about 488 mg to about 798 mg, about 532 mg to about 798 mg, about 576 mg to about 798 mg, about 621 mg to about 798 mg, about 665 mg to about 798 mg, about 709 mg to about 798 mg, about 753 mg to about 798 mg, about 22 mg to about 177 mg, about 44 mg to about 177 mg, about 44 mg to about 160 mg, about 44 mg to about 150 mg, about 44 mg to about 140 mg, about 44 mg to about 130 mg, about 44 mg to about 120 mg, about 44 mg to about 110 mg, about 44 mg to about 100 mg, about 44 mg to about 90 mg, about 44 mg to about 80 mg, about 44 mg to about 70 mg, about 44 mg to about 60 mg, or about 44 mg to about 50 mg.

[0206]

[0340] In various embodiments, as used herein, (i) about 22 mg to about 177 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof and an anti-angiogenic agent; (ii) one or more pharmaceutically acceptable excipients; and A pharmaceutical composition comprising:

[0207]

[0341] In various embodiments, as used herein, (i) about 44 mg to about 177 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof and an anti-angiogenic agent; (ii) one or more pharmaceutically acceptable excipients; and A pharmaceutical composition comprising:

[0208]

[0342] In various embodiments, as used herein, (i) about 177 mg to about 798 mg of a pharmaceutically acceptable salt of the compound of Formula (I) and an anti-angiogenic agent; (ii) one or more pharmaceutically acceptable excipients; and A pharmaceutical composition comprising:

[0209]

[0343] In another aspect, provided herein is a pharmaceutical composition comprising about 22 mg to about 798 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients, for the treatment of a cancer described herein in a subject in need of such treatment (e.g., bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer (e.g., malignant melanoma)), and thyroid cancer).

[0210]

[0344] In another aspect, provided herein is a pharmaceutical composition comprising about 22 mg to about 798 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients, for the treatment of a cancer described herein in a subject in need of such treatment (e.g., bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer), and thyroid cancer).

[0211]

[0345] In another aspect, provided herein is a pharmaceutical composition comprising about 177 mg to about 798 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients, for the treatment of a cancer described herein in a subject in need of such treatment (e.g., bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer (e.g., malignant melanoma)), and thyroid cancer).

[0212]

[0346] In another aspect, provided herein is a pharmaceutical composition comprising about 177 mg to about 798 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients, for the treatment of a cancer described herein in a subject in need of such treatment (e.g., bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer), and thyroid cancer).

[0213]

[0347] In another aspect, provided herein is a pharmaceutical composition comprising about 44 mg to about 177 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients, for the treatment of a cancer described herein in a subject in need of such treatment (e.g., bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer (e.g., malignant melanoma)), and thyroid cancer).

[0214]

[0348] In another aspect, provided herein is a pharmaceutical composition comprising about 44 mg to about 177 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients, for the treatment of a cancer described herein in a subject in need of such treatment (e.g., bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer), and thyroid cancer).

[0215]

[0349] In certain embodiments, a pharmaceutically acceptable salt of a compound of formula (I) is a fumarate salt. In certain embodiments, a pharmaceutically acceptable salt of a compound of formula (I) is a hemifumarate salt.

[0216]

[0350] The pharmaceutical compositions described herein can be administered by a variety of routes, including, but not limited to, oral (enteral), parenteral (by injection), rectal, transdermal, intradermal, intrathecal, subcutaneous (SC), intravenous (IV), intramuscular (IM), and intranasal administration. In certain embodiments, the pharmaceutical compositions described herein are administered orally.

[0217]

[0351] The pharmaceutical compositions described herein can be administered chronically ("chronic administration"). Chronic administration refers to administration of a compound or pharmaceutical composition thereof for an extended period of time, such as 3 months, 6 months, 1 year, 2 years, 3 years, 5 years, etc., or can be continued indefinitely, such as until the death of the subject. In certain embodiments, chronic administration is intended to provide a constant level of compound in the blood, for example, within a long-term therapeutic window.

[0218]

[0352] The pharmaceutical compositions described herein can be in the form of unit dosage forms to facilitate accurate dosage administration. The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined amount of active substance calculated to produce a desired therapeutic effect in association with suitable pharmaceutical excipients. Typical unit dosage forms include pre-filled, pre-measured ampoules or syringes for liquid compositions, or pills, tablets, capsules, etc. for solid compositions.

[0219]

[0353] In certain embodiments, the pharmaceutical compositions provided herein are administered to a patient as a solid dosage form, hi certain embodiments, the solid dosage form is a capsule.

[0220]

[0354] Although the description of pharmaceutical compositions provided herein primarily relates to pharmaceutical compositions suitable for administration to humans, one skilled in the art will understand that such compositions are generally suitable for administration to animals of all kinds. Modifications of pharmaceutical compositions suitable for administration to humans to make them suitable for administration to a variety of animals are well understood, and one skilled in the art of veterinary pharmacology can design and / or perform such modifications using routine experimentation. For a general discussion of the formulation and / or manufacture of pharmaceutical compositions, see, for example, Remington: The Science and Practice of Pharmacy 21 st ed., Lippincott Williams & Wilkins, 2005.

[0221] Uses and Treatment Methods

[0355] Provided herein are methods for activating the immune system of a subject in need thereof, generally comprising administering to the subject a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein, and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor).

[0222]

[0356] Also provided herein are methods for activating the immune system of a subject in need thereof, which generally comprise administering to the subject a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein, and a therapeutically effective amount of an anti-PD-1 antibody.

[0223]

[0357] Also provided herein is a method of activating the immune system of a subject having cancer (e.g., a cancer described herein), comprising administering to the subject a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein, and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor).

[0224]

[0358] Also provided herein is a method of activating the immune system of a subject with cancer (e.g., a cancer described herein), comprising administering to the subject a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein, and a therapeutically effective amount of an anti-PD-1 antibody.

[0225]

[0359] In some embodiments, the methods of activating the immune system described herein inhibit the activity of myeloid-derived suppressor cells. In some embodiments, the methods of activating the immune system described herein increase the infiltration of T cells and NK cells into tumors. In some embodiments, the methods of activating the immune system described herein increase the frequency of dendritic cells in draining lymph nodes.

[0226]

[0360] Also provided herein is a method for treating cancer in a subject in need thereof. The method generally comprises administering to the subject a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein, and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor). In certain embodiments, the cancer is a solid tumor.

[0227]

[0361] Also provided herein are methods of treating cancer in a subject in need thereof. The methods generally involve administering to the subject a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein, and a therapeutically effective amount of an anti-PD-1 antibody. In certain embodiments, the cancer is a solid tumor.

[0228]

[0362] The compounds of formula (I) or pharmaceutically acceptable salts thereof in combination with immunotherapy (e.g., anti-PD-1 antibodies) are useful in treating cancers including, but not limited to, pancreatic cancer; bladder cancer; colorectal cancer; breast cancer, including metastatic breast cancer; prostate cancer, including androgen-dependent and androgen-independent prostate cancer; kidney or renal cancer, including, for example, metastatic renal cell carcinoma; hepatocellular carcinoma; lung cancer, including, for example, NSCLC, bronchioloalveolar carcinoma (BAC), and lung adenocarcinoma; melanoma; neuroendocrine cancers, including metastatic neuroendocrine tumors; brain tumors, including glioma, anaplastic oligodendroglioma, adult glioblastoma multiforme, and adult anaplastic astrocytoma; bone cancer; and soft tissue sarcoma, including liver cancer, rectal cancer, penile cancer, vulvar cancer, thyroid cancer, salivary gland cancer, endometrial or uterine cancer, hepatoma, hepatocellular carcinoma, liver cancer, gastric cancer, including gastrointestinal cancer, peritoneal cancer, lung squamous cell carcinoma, gastroesophageal cancer, biliary tract cancer, gallbladder cancer, colorectal / appendix cancer, and squamous cell carcinoma (e.g., squamous cell carcinoma).

[0229]

[0363] In certain embodiments, the cancer is selected from the group consisting of bladder cancer, head and neck squamous cell carcinoma, malignant melanoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma.

[0230]

[0364] In certain embodiments, the cancer is selected from the group consisting of bladder cancer, head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma.

[0231]

[0365] In certain embodiments, the cancer is a liquid tumor. In certain embodiments, the cancer is selected from the group consisting of multiple myeloma, leukemia, and lymphoma. In certain embodiments, the leukemia is selected from the group consisting of chronic lymphocytic leukemia, chronic myelocytic leukemia, acute lymphoblastic leukemia, and acute myeloid leukemia (AML). In some embodiments, the lymphoma is selected from the group consisting of classical Hodgkin's lymphoma, primary thymic-mediastinal lymphoma, and non-Hodgkin's lymphoma. In certain embodiments, the lymphoma is selected from classical Hodgkin's lymphoma and primary thymic-mediastinal lymphoma.

[0232]

[0366] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof to a subject provides about 22 mg to about 226 mg, about 35 mg to about 226 mg, about 45 mg to about 226 mg, about 55 mg to about 226 mg, about 65 mg to about 226 mg, about 75 mg to about 226 mg, about 85 mg to about 226 mg, about 95 mg to about 226 mg, about 105 mg to about 226 mg, or about 125 mg to about 226 mg. Approximately 226 mg, approximately 115 mg to approximately 226 mg, approximately 125 mg to approximately 226 mg, approximately 135 mg to approximately 226 mg, approximately 145 mg to approximately 226 mg, approximately 155 mg to approximately 226 mg, approximately 165 mg ~ about 226mg, about 175mg - about 226mg, about 185mg - about 226mg, about 195mg - about 226mg, about 22mg - about 190mg, about 22mg - about 180mg, about 22 mg ~ about 170mg, about 22mg - about 160mg, about 22mg - about 150mg, about 22mg - about 140mg, about 22mg - about 130mg, about 22mg - about 120mg, about 22mg Approximately 110mg, approximately 22mg to approximately 100mg, approximately 22mg to approximately 90mg, approximately 22mg to approximately 80mg, approximately 22mg to approximately 70mg, approximately 22mg to approximately 60mg, approximately 22mg to approximately 50mg, approximately This includes administering 22 mg to about 40 mg, about 25 mg to about 30 mg, about 35 mg to about 185 mg, about 45 mg to about 175 mg, about 55 mg to about 165 mg, about 65 mg to about 155 mg, about 75 mg to about 145 mg, about 85 mg to about 135 mg, about 95 mg to about 125 mg, or about 105 mg to about 115 mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0233]

[0367] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof to a subject provides a therapeutically effective amount of about 177 mg to about 900 mg, about 250 mg to about 900 mg, about 300 mg to about 900 mg, about 350 mg to about 900 mg, about 400 mg to about 900 mg, about 450 mg to about 900 mg, about 500 mg to about 900 mg, about 550 mg to about 900 mg, about 600 mg to about 900 mg, about 650 mg to about 900 mg, about 700 mg to about 900 mg, about 750 mg to about 900 mg, about 800 mg to about 900 mg, about 850 mg to about 900 mg, about 177 mg to about 850 mg, about 177 mg to about 800 mg, about 177 mg about 177mg to about 750mg, about 177mg to about 700mg, about 177mg to about 650mg, about 177mg to about 600mg, about 177mg to about 550mg, about 177mg to about 500mg, about 177mg to about 450mg, about 177mg to about 400mg, about 177mg to about 350mg, about 177mg to about 300mg, about 177mg to about 250mg, about 250mg to about 850mg, about 300mg to about 800mg, about 350mg to about 750mg, about 400mg to about 700mg, about 450mg to about 650mg, about 500mg to about 600mg, or about 550mg to about 600mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0234]

[0368] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof to a subject provides about 22 mg to about 451 mg, about 50 mg to about 451 mg, about 75 mg to about 451 mg, about 100 mg to about 451 mg, about 125 mg to about 451 mg, about 150 mg to about 451 mg, about 175 mg to about 451 mg, about 200 mg to about 451 mg, about 225 mg to about 451 mg, about 250 mg to about 451 mg, about 265 mg to about 451 mg, about 275 mg to about 451 mg, about 285 mg to about 451 mg, about 300 mg to about 451 mg, about 325 mg to about 451 mg, about 345 mg to about 451 mg, about 365 mg to about 451 mg, about 385 mg to about 451 mg, about 395 mg to about 451 mg, about 400 mg to about 451 mg, about 415 mg to about 451 mg, about 425 mg to about 451 mg, about 435 mg to about 451 mg, about 445 mg to about 451 mg, about 451 mg to about 451 mg, about 465 mg to about 451 mg, about 475 mg to about 451 mg, about 485 mg to about 451 mg, about 495 mg to about 451 mg, about 500 mg to about 451 mg, about 525 mg to about 451 mg, about 545 mg to about 451 mg, about 565 mg to about 451 mg, about 585 mg to about 451 mg, about 595 mg to about Approximately 451 mg, approximately 250 mg to approximately 451 mg, approximately 275 mg to approximately 451 mg, approximately 300 mg to approximately 451 mg, approximately 325 mg to approximately 451 mg, approximately 350 mg to approximately 451 mg, approximately 375 mg ~451mg, 22mg~375mg, 22mg~350mg, 22mg~325mg, 22mg~300mg, 22mg~275mg, 22mg~250 mg, about 22 mg to about 225 mg, about 22 mg to about 200 mg, about 22 mg to about 175 mg, about 22 mg to about 150 mg, about 22 mg to about 125 mg, about 22 mg to about 100 mg, about 2 2mg to about 75mg, about 22mg to about 50mg, about 50mg to about 375mg, about 75mg to about 350mg, about 100mg to about 325mg, about 125mg to about 300mg, about 150mg to This includes administering about 275 mg, about 175 mg to about 250 mg, about 200 mg to about 225 mg, about 50 mg to about 100 mg, about 100 mg to about 150 mg, about 150 mg to about 200 mg, about 200 mg to about 250 mg, about 250 mg to about 300 mg, about 300 mg to about 350 mg, or about 350 mg to about 451 mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0235]

[0369] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof provides a subject with a therapeutically effective amount of about 22 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, about 115 mg, about 120 mg, about 125 mg, about 130 mg, about 135 mg, about 140 mg , about 145 mg, about 150 mg, about 155 mg, about 160 mg, about 165 mg, about 170 mg, about 175 mg, about 180 mg, about 185 mg, about 190 mg, about 195 mg, about 200 mg, about 210 mg, about 220 mg, about 2 30mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg, about 300mg, about 310mg, about 320mg, about 330mg, about 340mg, about 350mg, about 360mg, about 370mg , about 380mg, about 390mg, about 400mg, about 410mg, about 420mg, about 430mg, about 440mg, about 450mg, about 460mg, about 470mg, about 480mg, about 490mg, about 500mg, about 510mg, about 520mg, about 530mg, about 540mg, about 550mg, about 560mg, about 570mg, about 580mg, about 590mg, about 600mg, about 610mg, about 620mg, about 630mg, about 640mg, about 650mg, about 660mg , about 670 mg, about 680 mg, about 690 mg, about 700 mg, about 710 mg, about 720 mg, about 730 mg, about 740 mg, about 750 mg, about 760 mg, about 770 mg, about 780 mg, about 790 mg, about 800 mg, about 810 mg, about 820 mg, about 830 mg, about 840 mg, about 850 mg, about 860 mg, about 870 mg, about 880 mg, about 890 mg or about 900 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0236]

[0370] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 22 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 50 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 75 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 100 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 125 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 150 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 175 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 200 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 250 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 300 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 350 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 400 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 450 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 500 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 550 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 600 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 650 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 700 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 750 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 800 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 850 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 900 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

[0237]

[0371] In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) to a subject provides a therapeutically effective amount of about 22 mg to about 226 mg, about 35 mg to about 226 mg, about 45 mg to about 226 mg, about 55 mg to about 226 mg, about 65 mg to about 226 mg, about 75 mg to about 226 mg, about 85 mg to about 226 mg, about 95 mg to about 226 mg, about 105 mg to about 226 mg, or about 125 mg to about 226 mg. 26mg, about 115mg to about 226mg, about 125mg to about 226mg, about 135mg to about 226mg, about 145mg to about 226mg, about 155mg to about 226mg, about 165m g ~ about 226mg, about 175mg - about 226mg, about 185mg - about 226mg, about 195mg - about 226mg, about 22mg - about 190mg, about 22mg - about 180mg, about 22 mg ~ about 170mg, about 22mg - about 160mg, about 22mg - about 150mg, about 22mg - about 140mg, about 22mg - about 130mg, about 22mg - about 120mg, about 22mg ~110mg, 22mg~100mg, 22mg~90mg, 22mg~80mg, 22mg~70mg, 22mg~60mg, 22mg~50mg , about 22 mg to about 40 mg, about 22 mg to about 30 mg, about 35 mg to about 185 mg, about 45 mg to about 175 mg, about 55 mg to about 165 mg, about 65 mg to about 155 mg, about 75 mg to about 145 mg, about 85 mg to about 135 mg, about 95 mg to about 125 mg, or about 105 mg to about 115 mg of a pharmaceutically acceptable salt of the compound of formula (I).

[0238]

[0372] In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) to a subject provides a therapeutically effective amount of about 177 mg to about 900 mg, about 250 mg to about 900 mg, about 300 mg to about 900 mg, about 350 mg to about 900 mg, about 400 mg to about 900 mg, about 450 mg to about 900 mg, about 500 mg to about 900 mg, about 550 mg to about 900 mg, about 600 mg to about 900 mg, about 650 mg to about 900 mg, about 700 mg to about 900 mg, about 750 mg to about 900 mg, about 800 mg to about 900 mg, about 850 mg to about 900 mg, about 177 mg to about 850 mg, about 177 mg to about 800 mg, about 177 mg about 177mg to about 750mg, about 177mg to about 700mg, about 177mg to about 650mg, about 177mg to about 600mg, about 177mg to about 550mg, about 177mg to about 500mg, about 177mg to about 450mg, about 177mg to about 400mg, about 177mg to about 350mg, about 177mg to about 300mg, about 177mg to about 250mg, about 250mg to about 850mg, about 300mg to about 800mg, about 350mg to about 750mg, about 400mg to about 700mg, about 450mg to about 650mg, about 500mg to about 600mg, or about 550mg to about 600mg of a pharmaceutically acceptable salt of the compound of formula (I).

[0239]

[0373] In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) to a subject provides about 22 mg to about 451 mg, about 50 mg to about 451 mg, about 75 mg to about 451 mg, about 100 mg to about 451 mg, about 125 mg to about 451 mg, about 150 mg to about 451 mg, about 175 mg to about 451 mg, about 200 mg to about 451 mg, about 225 mg to about 451 mg, or about 300 mg to about 451 mg. 451mg, about 250mg to about 451mg, about 275mg to about 451mg, about 300mg to about 451mg, about 325mg to about 451mg, about 350mg to about 451mg, about 375mg ~451mg, 22mg~375mg, 22mg~350mg, 22mg~325mg, 22mg~300mg, 22mg~275mg, 22mg~250 mg, about 22mg to about 225mg, about 22mg to about 200mg, about 22mg to about 175mg, about 22mg to about 150mg, about 22mg to about 125mg, about 22mg to about 100mg, about 22mg to about 75mg, about 22mg to about 50mg, about 50mg to about 375mg, about 75mg to about 350mg, about 100mg to about 325mg, about 125mg to about 300mg, about 150m The method includes administering from about 100 mg to about 275 mg, from about 175 mg to about 250 mg, from about 200 mg to about 225 mg, from about 50 mg to about 100 mg, from about 100 mg to about 150 mg, from about 150 mg to about 200 mg, from about 200 mg to about 250 mg, from about 250 mg to about 300 mg, from about 300 mg to about 350 mg, or from about 350 mg to about 451 mg of a pharmaceutically acceptable salt of the compound of formula (I).

[0240]

[0374] In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) provides a subject with a therapeutically effective amount of about 22 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, about 115 mg, about 120 mg, about 125 mg, about 130 mg, about 135 mg, about 140 mg, about 145mg, about 150mg, about 155mg, about 160mg, about 165mg, about 170mg, about 175mg, about 180mg, about 185mg, about 190mg, about 195mg, about 200mg, about 210mg, about 220mg, about 23 0mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg, about 300mg, about 310mg, about 320mg, about 330mg, about 340mg, about 350mg, about 360mg, about 370mg , about 380mg, about 390mg, about 400mg, about 410mg, about 420mg, about 430mg, about 440mg, about 450mg, about 460mg, about 470mg, about 480mg, about 490mg, about 500mg, about 510mg, about 520mg, about 530mg, about 540mg, about 550mg, about 560mg, about 570mg, about 580mg, about 590mg, about 600mg, about 610mg, about 620mg, about 630mg, about 640mg, about 650mg, about 660 The method comprises administering about 670 mg, about 680 mg, about 690 mg, about 700 mg, about 710 mg, about 720 mg, about 730 mg, about 740 mg, about 750 mg, about 760 mg, about 770 mg, about 780 mg, about 790 mg, about 800 mg, about 810 mg, about 820 mg, about 830 mg, about 840 mg, about 850 mg, about 860 mg, about 870 mg, about 880 mg, about 890 mg or about 900 mg of a pharmaceutically acceptable salt of a compound of Formula (I).

[0241]

[0375] In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) comprises administering to the subject about 22 mg of a pharmaceutically acceptable salt of a compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) comprises administering to the subject about 50 mg of a pharmaceutically acceptable salt of a compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) comprises administering to the subject about 75 mg of a pharmaceutically acceptable salt of a compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) comprises administering to the subject about 100 mg of a pharmaceutically acceptable salt of a compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) comprises administering to the subject about 125 mg of a pharmaceutically acceptable salt of a compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) comprises administering to the subject about 150 mg of a pharmaceutically acceptable salt of a compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) comprises administering to the subject about 175 mg of a pharmaceutically acceptable salt of a compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) comprises administering to the subject about 200 mg of a pharmaceutically acceptable salt of a compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) comprises administering to the subject about 250 mg of a pharmaceutically acceptable salt of a compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) comprises administering to the subject about 300 mg of a pharmaceutically acceptable salt of a compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of the compound of Formula (I) comprises administering to the subject about 350 mg of a pharmaceutically acceptable salt of the compound of Formula (I).In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of the compound of Formula (I) comprises administering to the subject about 400 mg of a pharmaceutically acceptable salt of the compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of the compound of Formula (I) comprises administering to the subject about 450 mg of a pharmaceutically acceptable salt of the compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of the compound of Formula (I) comprises administering to the subject about 500 mg of a pharmaceutically acceptable salt of the compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of the compound of Formula (I) comprises administering to the subject about 550 mg of a pharmaceutically acceptable salt of the compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of the compound of Formula (I) comprises administering to the subject about 600 mg of a pharmaceutically acceptable salt of the compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of the compound of Formula (I) comprises administering to the subject about 650 mg of a pharmaceutically acceptable salt of the compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of the compound of Formula (I) comprises administering to the subject about 700 mg of a pharmaceutically acceptable salt of the compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of the compound of Formula (I) comprises administering to the subject about 750 mg of a pharmaceutically acceptable salt of the compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of the compound of Formula (I) comprises administering to the subject about 800 mg of a pharmaceutically acceptable salt of the compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of the compound of Formula (I) comprises administering to the subject about 850 mg of a pharmaceutically acceptable salt of the compound of Formula (I). In some embodiments, administering a therapeutically effective amount of a pharmaceutically acceptable salt of the compound of Formula (I) comprises administering to the subject about 900 mg of a pharmaceutically acceptable salt of the compound of Formula (I).

[0242]

[0376] In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject. In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject daily. In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily. In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily.

[0243]

[0377] In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 consecutive days. In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for 21 consecutive days.

[0244]

[0378] In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 consecutive days. In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for at least 21 consecutive days.

[0245]

[0379] In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 consecutive days. In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for 21 consecutive days.

[0246]

[0380] In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 consecutive days. In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for at least 21 consecutive days.

[0247]

[0381] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, or 24 months. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for 1 year. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for 2 years.

[0248]

[0382] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for at least 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, or 24 months. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for at least 1 year. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for at least 2 years.

[0249]

[0383] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, or 24 months. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for 1 year. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for 2 years.

[0250]

[0384] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for at least 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, or 24 months. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for at least 1 year. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for at least 2 years.

[0251]

[0385] In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject from about 22 mg to about 451 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

[0252]

[0386] In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject between about 22 mg and about 451 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

[0253]

[0387] In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject from about 22 mg to about 451 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof daily.

[0254]

[0388] In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject from about 22 mg to about 451 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof once daily.

[0255]

[0389] In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject between about 22 mg and about 451 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof twice daily.

[0256]

[0390] In certain embodiments, the subject is fasting. In certain embodiments, the subject is not fasting. In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering an effective amount to the subject within about 30 minutes of completing a meal.

[0257]

[0391] In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 22 mg to about 451 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof within about 30 minutes of completing a meal.

[0258]

[0392] In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject between about 177 mg and about 900 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

[0259]

[0393] In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject between about 177 mg and about 900 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

[0260]

[0394] In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject from about 177 mg to about 900 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof daily.

[0261]

[0395] In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject from about 177 mg to about 900 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof once daily.

[0262]

[0396] In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject between about 177 mg and about 900 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof twice daily.

[0263]

[0397] In certain embodiments, the subject is fasting. In certain embodiments, the subject is not fasting. In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering a therapeutically effective amount to the subject within about 30 minutes of completing a meal.

[0264]

[0398] In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject about 177 mg to about 900 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof within about 30 minutes of completing a meal.

[0265]

[0399] In some embodiments, the immunotherapy is selected from the group consisting of an immune checkpoint inhibitor, an adoptive cellular therapy, and a monoclonal antibody. In some embodiments, the immunotherapy is selected from an immune checkpoint inhibitor and an adoptive cellular therapy.

[0266]

[0400] In certain embodiments, the immunotherapy is adoptive cell therapy, hi certain embodiments, the adoptive cell therapy is selected from the group consisting of tumor infiltrating lymphocyte therapy, engineered T cell receptor therapy, CAR T cell therapy, and natural killer cell therapy.

[0267]

[0401] In some embodiments, the immunotherapy is an immune checkpoint inhibitor, hi some embodiments, the immune checkpoint inhibitor is a PD-1 (programmed cell death protein 1) antagonist, a PD-L1 (programmed cell death protein ligand 1) antagonist, a PD-L2 antagonist, a CD27 (cluster of differentiation 27) antagonist, a CD28 antagonist, a CD70 antagonist, a CD80 antagonist, a CD86 antagonist, a CD137 antagonist, a CD276 antagonist, a KIR (killer cell immunoglobulin-like receptor) antagonist, a LAG3 (lymphocyte activation gene 3) antagonist, a TNFRSF4 (tumor necrosis factor receptor superfamily, member 4) antagonist, or a GITR (glucocorticoid-inducible TNFR-related protein) antagonist. , GITRL (GITR ligand) antagonists, 4-1BBL (4-1BB ligand) antagonists, CTLA-4 (cytotoxic T lymphocyte-associated antigen 4) antagonists, A2AR (adenosine A2A receptor) antagonists, VTCN1 (V-set domain-containing T-cell activation inhibitor 1) antagonists, BTLA (B and T lymphocyte attenuator) antagonists, IDO (indoleamine 2,3-dioxygenase) antagonists, TIM-3 (T cell immunoglobulin and mucin domain-containing-3) antagonists, VISTA (V-domain Ig suppressor of T cell activation) antagonists, KLRA antagonists, and combinations thereof.

[0268]

[0402] In some embodiments, the immune checkpoint inhibitor is ipilimumab, tremelimumab, nivolumab, pembrolizumab, pidilizumab, lambrolizumab, BMS-936559, atezolizumab, AMP-224, AMP224, AUNP12, BGB108, MCLA134, MEDI0680, spartalizumab, cemiplimab, SHR1210, ST IA110X, STIA1110, TSR042, MPDL3280A, MEdi-4736, MSB0010718C, ALN-PDL, BGBA317, KD033, KY1003, STIA100X, STIA1010, STIA1011, STIA1012, STIA101, BMS-663513 and PF-05082566.

[0269]

[0403] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapy are administered simultaneously. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapy are administered separately. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapy are administered sequentially.

[0270]

[0404] In some embodiments, a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) and a therapeutically effective amount of an immunotherapy are administered simultaneously. In some embodiments, a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) and a therapeutically effective amount of an immunotherapy are administered separately. In some embodiments, a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) and a therapeutically effective amount of an immunotherapy are administered sequentially.

[0271]

[0405] In some embodiments, a therapeutically effective amount of immunotherapy is administered over 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, or 24 months. In some embodiments, a therapeutically effective amount of immunotherapy is administered over 1 year. In some embodiments, a therapeutically effective amount of immunotherapy is administered over 2 years.

[0272]

[0406] In some embodiments, the immune checkpoint inhibitor is an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody. In some embodiments, the immune checkpoint inhibitor is an anti-PD-1 antibody.

[0273]

[0407] In some embodiments, the anti-PD-1 antibody is a PD-1 antagonist. In some embodiments, the anti-PD-1 antibody is a PD-1 agonist. In some embodiments, the anti-PD-1 antibody is a PD-1 inhibitor.

[0274]

[0408] In some embodiments, the anti-PD-1 antibody is selected from the group consisting of nivolumab, pembrolizumab, spartalizumab, tislelizumab, or cemiplimab. In some embodiments, the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, and cemiplimab. In certain embodiments, the anti-PD-1 antibody is selected from pembrolizumab and nivolumab.

[0275]

[0409] In certain embodiments, the anti-PD-1 antibody is pembrolizumab. In certain embodiments, the anti-PD-1 antibody is nivolumab. In certain embodiments, the anti-PD-1 antibody is cemiplimab.

[0276]

[0410] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody are administered simultaneously. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody are administered separately. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody are administered sequentially.

[0277]

[0411] In some embodiments, a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) and a therapeutically effective amount of an anti-PD-1 antibody are administered simultaneously. In some embodiments, a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) and a therapeutically effective amount of an anti-PD-1 antibody are administered separately. In some embodiments, a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of Formula (I) and a therapeutically effective amount of an anti-PD-1 antibody are administered sequentially.

[0278]

[0412] In some embodiments, a therapeutically effective amount of an anti-PD-1 antibody is administered for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, or 24 months. In some embodiments, a therapeutically effective amount of an anti-PD-1 antibody is administered for 1 year. In some embodiments, a therapeutically effective amount of an anti-PD-1 antibody is administered for 2 years.

[0279]

[0413] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody are administered over 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, or 24 months. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody are administered over 1 year. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody are administered over 2 years.

[0280]

[0414] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody are administered for at least 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, or 24 months. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody are administered for at least 1 year. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody are administered for at least 2 years.

[0281]

[0415] In some embodiments, administering a therapeutically effective amount of an anti-PD-1 antibody comprises administering about 200 mg to about 400 mg of the anti-PD-1 antibody to the subject. In some embodiments, administering a therapeutically effective amount of an anti-PD-1 antibody comprises administering about 200 mg to about 400 mg of the anti-PD-1 antibody to the subject about every 2 to 4 weeks. In some embodiments, administering a therapeutically effective amount of an anti-PD-1 antibody comprises administering about 200 mg to about 400 mg of the anti-PD-1 antibody to the subject about every 2 to 4 weeks for about 6 months. In some embodiments, administering a therapeutically effective amount of an anti-PD-1 antibody comprises administering about 200 mg to about 400 mg of the anti-PD-1 antibody to the subject about every 2 to 4 weeks for about 1 year. In some embodiments, administering a therapeutically effective amount of an anti-PD-1 antibody comprises administering about 200 mg to about 400 mg of the anti-PD-1 antibody to the subject about every 2 to 4 weeks for about 2 years.

[0282]

[0416] In some embodiments, the anti-PD-1 antibody is pembrolizumab. In certain embodiments, pembrolizumab is administered in an amount of about 200 mg every three weeks. In certain embodiments, pembrolizumab is administered in an amount of about 200 mg every three weeks for about six months. In certain embodiments, pembrolizumab is administered in an amount of about 200 mg every three weeks for about one year. In certain embodiments, pembrolizumab is administered in an amount of about 200 mg every three weeks for about two years. In certain embodiments, pembrolizumab is administered in an amount of about 400 mg every six weeks. In certain embodiments, pembrolizumab is administered in an amount of about 400 mg every six weeks for about six months. In certain embodiments, pembrolizumab is administered in an amount of about 400 mg every six weeks for about one year. In certain embodiments, pembrolizumab is administered in an amount of about 400 mg every six weeks for about two years.

[0283]

[0417] In some embodiments, the anti-PD-1 antibody is nivolumab. In certain embodiments, nivolumab is administered in an amount of about 240 mg every two weeks. In certain embodiments, nivolumab is administered in an amount of about 240 mg every two weeks for about six months. In certain embodiments, nivolumab is administered in an amount of about 240 mg every two weeks for about one year. In certain embodiments, nivolumab is administered in an amount of about 240 mg every two weeks for about two years. In certain embodiments, nivolumab is administered in an amount of about 480 mg every four weeks. In certain embodiments, nivolumab is administered in an amount of about 480 mg every four weeks for about six months. In certain embodiments, nivolumab is administered in an amount of about 480 mg every four weeks for about one year. In certain embodiments, nivolumab is administered in an amount of about 480 mg every four weeks for about two years.

[0284]

[0418] In some embodiments, the anti-PD-1 antibody is cemiplimab. In certain embodiments, cemiplimab is administered at an amount of about 350 mg every three weeks. In certain embodiments, cemiplimab is administered at an amount of about 350 mg every three weeks for about six months. In certain embodiments, cemiplimab is administered at an amount of about 350 mg every three weeks for about one year. In certain embodiments, cemiplimab is administered at an amount of about 350 mg every three weeks for about two years.

[0285]

[0419] In some embodiments, provided herein is a method of treating cancer (e.g., a cancer described herein) in a subject in need of such treatment, comprising administering to the subject a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, as described herein; or a pharmaceutical composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof, as described herein; and a therapeutically effective amount of pembrolizumab.

[0286]

[0420] In some embodiments, provided herein is a method of treating cancer (e.g., a cancer described herein) in a subject in need of such treatment, comprising administering to the subject a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, as described herein; or a pharmaceutical composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof, as described herein; and a therapeutically effective amount of nivolumab.

[0287]

[0421] In some embodiments, provided herein is a method of treating cancer (e.g., a cancer described herein) in a subject in need of such treatment, comprising administering to the subject a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, as described herein; or a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, as described herein; and a therapeutically effective amount of cemiplimab.

[0288]

[0422] In certain embodiments, the subject has received at least one line of prior therapy. In certain embodiments, the subject has received fewer than five lines of prior therapy. In certain embodiments, the subject has received one, two, three, or four lines of prior therapy. In certain embodiments, the subject has received no prior therapy.

[0289]

[0423] In certain embodiments, the subject has received at least one but no more than five lines of prior therapy.

[0290]

[0424] Prior treatments include, but are not limited to, surgery, radiation therapy (e.g., external beam radiation therapy or internal radiation therapy), chemotherapy (e.g., alkylating agents, nitrosoureas, antimetabolites, plant alkaloids and natural products, antitumor antibiotics, hormones and biological response modifiers), gene therapy, DNA therapy, viral therapy (e.g., oncolytic virus therapy), RNA therapy, adjuvant therapy, and immunotherapy (e.g., immune checkpoint inhibition, adoptive cell therapy, (e.g., tumor-infiltrating lymphocyte therapy, modified T-cell receptor therapy, CAR T-cell therapy, natural killer cell therapy), or monoclonal antibodies).

[0291]

[0425] In certain embodiments, the method includes administering a therapeutically effective amount of a pharmaceutically acceptable salt of the compound of formula (I). In certain embodiments, the pharmaceutically acceptable salt is a fumarate salt. In certain embodiments, the fumarate salt is a hemifumarate salt.

[0292]

[0426] In certain embodiments, the methods described herein further comprise administering to the subject a therapeutically effective amount of a third therapeutic agent, hi certain embodiments, the second therapeutic agent is selected from the group consisting of a checkpoint inhibitor, an EGFR inhibitor, an antiangiogenic agent, venetoclax, fluorouracil, and combinations thereof.

[0293]

[0427] In certain embodiments, the third therapeutic agent is a checkpoint inhibitor. In certain embodiments, the third therapeutic agent is a PD-L1 inhibitor. In certain embodiments, the third therapeutic agent is selected from the group consisting of atezolizumab, avelumab, and durvalumab.

[0294]

[0428] In certain embodiments, the third therapeutic agent is an EGFR inhibitor, hi some embodiments, the EGFR inhibitor is selected from the group consisting of erlotinib, gefitinib, afatinib, and osimertinib.

[0295]

[0429] In certain embodiments, the third therapeutic agent is an anti-angiogenic agent. In certain embodiments, the anti-angiogenic agent is a VEGFR inhibitor. In certain embodiments, the anti-angiogenic agent is a VEGFR tyrosine kinase inhibitor.

[0296]

[0430] In certain embodiments, the third therapeutic agent is a VEGFR inhibitor, hi certain embodiments, the VEGFR inhibitor is selected from the group consisting of sunitinib, axitinib, lenvatinib, tivozanib, pazopanib, cabozantinib, and ramucirumab.

[0297]

[0431] In another aspect, provided herein is a method of treating cancer (e.g., a cancer described herein) in a subject in need thereof, the method comprising administering to the subject any one of the pharmaceutical compositions described herein and a therapeutically effective amount of an anti-PD-1 antibody (e.g., an anti-PD-1 antibody described herein).

[0298]

[0432] In another aspect, provided herein is a method of activating the immune system of a subject in need thereof, the method comprising administering to the subject any one of the pharmaceutical compositions described herein and a therapeutically effective amount of an anti-PD-1 antibody (e.g., an anti-PD-1 antibody described herein).

[0299]

[0433] In another aspect, provided herein is a method of activating the immune system of a subject having cancer (e.g., a cancer described herein), the method comprising administering to the subject any one of the pharmaceutical compositions described herein and a therapeutically effective amount of an anti-PD-1 antibody (e.g., an anti-PD-1 antibody described herein).

[0300]

[0434] In some embodiments, the methods described herein increase tumor cell expression of type 1 interferon receptor (INFAR1) in a subject compared to a subject receiving a placebo, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of an anti-PD-1 antibody.

[0301]

[0435] In some embodiments, the methods described herein increase peripheral blood monocyte surface expression of INFAR1 in a subject compared to a subject receiving a placebo, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of an anti-PD-1 antibody.

[0302]

[0436] In some embodiments, the methods described herein increase tumor cell expression of calreticulin in a subject compared to a subject receiving a placebo, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of an anti-PD-1 antibody.

[0303]

[0437] In some embodiments, the methods described herein increase polymorphonuclear myeloid-derived suppressor cell (MDSC) and / or tumor-associated macrophage (TAM) expression of INFAR1 in a subject compared to a subject receiving a placebo, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of an anti-PD-1 antibody.

[0304]

[0438] In some embodiments, the methods described herein increase TAM expression of PD-L1 in a subject relative to a subject receiving a placebo, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of an anti-PD-1 antibody.

[0305]

[0439] In some embodiments, the methods described herein increase the infiltration of CD8 T cells and / or NK cells into tumors in a subject compared to a subject receiving a placebo, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of an anti-PD-1 antibody.

[0306]

[0440] In some embodiments, the methods described herein increase expression of CD69 on T cells in tumor-draining lymph nodes in a subject compared to a subject receiving a placebo, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of an anti-PD-1 antibody.

[0307]

[0441] In some embodiments, the methods described herein reduce the suppressive activity of MDSCs in a subject.

[0308]

[0442] In some embodiments, the methods described herein increase the frequency of dendritic cells in tumor-draining lymph nodes in a subject.

[0309]

[0443] In some embodiments, the methods described herein increase Ki67, GzmB and memory phenotype markers on CD4 T cells in a subject.

[0310]

[0444] In some embodiments, the methods described herein increase macrophages of an immunostimulatory phenotype in the tumor microenvironment.

[0311]

[0445] In some embodiments, the methods described herein reduce the immunosuppressive phenotype of macrophages in the tumor microenvironment.

[0312]

[0446] In some embodiments, the methods described herein increase macrophages of M1 phenotype and / or function or decrease macrophages of M2 phenotype or function.

[0313]

[0447] In some embodiments, the methods described herein increase the surface expression of activation markers (eg, MHCII, HLA-DR and / or CD86) on macrophages or other myeloid cells.

[0314]

[0448] In some embodiments, the methods described herein result in an expansion of dendritic cells of an immunostimulatory phenotype in the draining lymph node or tumor microenvironment.

[0315]

[0449] In some embodiments, provided herein is a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of immunotherapy for use in treating cancer (e.g., a cancer described herein).

[0316]

[0450] In some embodiments, provided herein is a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody for use in treating cancer (e.g., a cancer described herein).

[0317]

[0451] In some embodiments, provided herein is a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of immunotherapy for use in activating the immune system of a subject in need thereof (e.g., a subject having a cancer described herein).

[0318]

[0452] In some embodiments, provided herein is a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-PD-1 antibody for use in activating the immune system of a subject in need thereof (e.g., a subject having a cancer described herein).

[0319]

[0453] Also provided herein is a method for treating cancer in a subject in need thereof. The method generally comprises administering to the subject a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutical composition described herein and a therapeutically effective amount of an anti-angiogenic agent (e.g., a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI)). In certain embodiments, the cancer is a solid tumor.

[0320]

[0454] Also provided herein is a method for treating cancer in a subject in need thereof. The method generally comprises administering to the subject a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutical composition described herein, a therapeutically effective amount of an anti-angiogenic agent (e.g., a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI)), and a therapeutically effective amount of an immunotherapy (e.g., an immune checkpoint inhibitor). In certain embodiments, the cancer is a solid tumor.

[0321]

[0455] It is contemplated that the compounds of formula (I) or pharmaceutically acceptable salts thereof in combination with an anti-angiogenic agent (e.g., a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI)) are useful in the treatment of cancer, including, but not limited to, bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer (e.g., malignant melanoma)) and thyroid cancer.

[0322]

[0456] It is contemplated that the compounds of formula (I) or pharmaceutically acceptable salts thereof in combination with an anti-angiogenic agent (e.g., a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI)) are useful in the treatment of cancer, including, but not limited to, bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer) and thyroid cancer.

[0323]

[0457] In certain embodiments, the cancer is selected from the group consisting of bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer (e.g., malignant melanoma)), and thyroid cancer.

[0324]

[0458] In certain embodiments, the cancer is selected from the group consisting of bladder cancer, breast cancer (e.g., triple-negative breast cancer or metastatic breast cancer), carcinoma (e.g., carcinoma of unknown primary (CUP), endometrial cancer, Merkel cell carcinoma, squamous cell carcinoma, or urothelial carcinoma), cervical cancer, colorectal cancer, hepatocellular carcinoma, gastric cancer (e.g., adenocarcinoma or gastrointestinal stromal tumor), renal cancer (e.g., renal cell carcinoma (RCC) such as renal clear cell carcinoma (ccRCC)), lung cancer (e.g., small cell lung cancer (SCLC) or non-small cell lung cancer), neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer), and thyroid cancer.

[0325]

[0459] In some embodiments, the cancer is malignant melanoma.

[0326]

[0460] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof to a subject provides a therapeutically effective amount of about 22 mg to about 798 mg, about 35 mg to about 798 mg, about 44 mg to about 798 mg, about 66 mg to about 798 mg, about 88 mg to about 798 mg, about 110 mg to about 798 mg, about 113 mg to about 798 mg, about 155 mg to about 798 mg, about 177 mg to about 798 mg, about 199 mg to about 798 mg, about 221 mg to about 798 mg, about 244 mg to about 798 mg, about 266 mg to about 798 mg, about 310 mg to about 798 mg, about 355 mg to about 798 mg, about 399 mg to about 798 mg, about 443 mg to about 798 mg, or about 488 mg to about 798 mg , about 532 mg to about 798 mg, about 576 mg to about 798 mg, about 621 mg to about 798 mg, about 665 mg to about 798 mg, about 709 mg to about 798 mg, about 753 mg to about 798 mg, about 22 mg to about 177 mg, about 44 mg to about 177 mg, about 44 mg to about 160 mg, about 44 mg to about 150 mg, about 44 mg to about 140 mg, about 44 mg to about 130 mg, about 44 mg to about 120 mg, about 44 mg to about 110 mg, about 44 mg to about 100 mg, about 44 mg to about 90 mg, about 44 mg to about 80 mg, about 44 mg to about 70 mg, about 44 mg to about 60 mg, or about 44 mg to about 50 mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0327]

[0461] In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject. In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject daily. In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily. In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily.

[0328]

[0462] In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 consecutive days. In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for 21 consecutive days.

[0329]

[0463] In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 consecutive days. In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for at least 21 consecutive days.

[0330]

[0464] In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 consecutive days. In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for 21 consecutive days.

[0331]

[0465] In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 consecutive days. In certain embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for at least 21 consecutive days.

[0332]

[0466] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, or 24 months. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for 1 year. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for 2 years.

[0333]

[0467] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for at least 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, or 24 months. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for at least 1 year. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for at least 2 years.

[0334]

[0468] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, or 24 months. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for 1 year. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for 2 years.

[0335]

[0469] In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject twice daily for at least 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, or 24 months. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for at least 1 year. In some embodiments, a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a subject once daily for at least 2 years.

[0336]

[0470] In certain embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an antiangiogenic agent (e.g., a VEGFR-TKI) comprises administering to the subject about 22 mg to about 798 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0337]

[0471] In certain embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an antiangiogenic agent (e.g., a VEGFR-TKI) comprises orally administering to the subject about 22 mg to about 798 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0338]

[0472] In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an antiangiogenic agent (e.g., a VEGFR-TKI) comprises orally administering to the subject about 22 mg to about 798 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof daily.

[0339]

[0473] In certain embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an antiangiogenic agent (e.g., a VEGFR-TKI) comprises orally administering to a subject about 22 mg to about 798 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof once daily.

[0340]

[0474] In certain embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an antiangiogenic agent (e.g., a VEGFR-TKI) comprises orally administering to a subject about 22 mg to about 798 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof twice daily.

[0341]

[0475] In certain embodiments, the subject is fasting. In certain embodiments, the subject is not fasting. In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-angiogenic agent (e.g., a VEGFR-TKI) comprises administering the therapeutically effective amounts to the subject within about 30 minutes of completing a meal.

[0342]

[0476] In certain embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an antiangiogenic agent (e.g., a VEGFR-TKI) comprises administering to the subject about 22 mg to about 798 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof within about 30 minutes of completing a meal.

[0343]

[0477] In certain embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an antiangiogenic agent (e.g., a VEGFR-TKI) comprises administering to the subject about 177 mg to about 798 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0344]

[0478] In certain embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an antiangiogenic agent (e.g., a VEGFR-TKI) comprises orally administering to the subject about 177 mg to about 798 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0345]

[0479] In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an antiangiogenic agent (e.g., a VEGFR-TKI) comprises orally administering to the subject about 177 mg to about 798 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof daily.

[0346]

[0480] In certain embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an antiangiogenic agent (e.g., a VEGFR-TKI) comprises orally administering to a subject about 177 mg to about 798 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof once daily.

[0347]

[0481] In certain embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an antiangiogenic agent (e.g., a VEGFR-TKI) comprises orally administering to a subject about 177 mg to about 798 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof twice daily.

[0348]

[0482] In certain embodiments, the subject is fasting. In certain embodiments, the subject is not fasting. In certain embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-angiogenic agent (e.g., a VEGFR-TKI) comprises administering the therapeutically effective amounts to the subject within about 30 minutes of completing a meal.

[0349]

[0483] In certain embodiments, administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an antiangiogenic agent (e.g., a VEGFR-TKI) comprises administering about 177 mg to about 798 mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof to a subject within about 30 minutes of completing a meal.

[0350]

[0484] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject from about 22 mg to about 798 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof on a free base equivalent weight basis.

[0351]

[0485] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject from about 22 mg to about 798 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof on a free base equivalent weight basis.

[0352]

[0486] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject from about 22 mg to about 798 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof daily on a free base equivalent weight basis.

[0353]

[0487] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject from about 22 mg to about 798 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, on a free base equivalent weight basis, once daily.

[0354]

[0488] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject from about 22 mg to about 798 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, on a free base equivalent weight basis, twice daily.

[0355]

[0489] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject from about 44 mg to about 177 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof on a free base equivalent weight basis.

[0356]

[0490] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject from about 44 mg to about 177 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof on a free base equivalent weight basis.

[0357]

[0491] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject from about 44 mg to about 177 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof daily, on a free base equivalent weight basis.

[0358]

[0492] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject from about 44 mg to about 177 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, on a free base equivalent weight basis, once daily.

[0359]

[0493] In some embodiments, administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to a subject from about 44 mg to about 177 mg of a compound of Formula (I) or a pharmaceutically acceptable salt thereof on a free base equivalent weight basis twice daily.

[0360]

[0494] In certain embodiments, the subject is a human. In certain embodiments, the subject is an adult human. [Example]

[0361] Example

[0495] In order that the present disclosure may be better understood, the following examples are set forth: The synthetic and biological examples described herein are presented to illustrate the compounds, pharmaceutical compositions and methods provided herein and should not be construed in any way as limiting the scope thereof.

[0362] Abbreviation BID twice a day BIW twice a week DMEM Dulbecco's Modified Eagle's Medium DMSO dimethyl sulfoxide dppf 1,1'-bis(diphenylphosphino)ferrocene EtOH ethanol FBS Fetal Bovine Serum h time HPLC High-Performance Liquid Chromatography IMDM Iscove's Modified Dulbecco's Medium IP intraperitoneal IPA Isopropyl Alcohol LN lymph node LOQ Limit of Quantification mAb monoclonal antibody MEM Minimum Essential Medium NEAA Non-Essential Amino Acids NMR nuclear magnetic resonance spectroscopy PBS Phosphate-buffered saline PO Oral SEM Standard error of the mean SFC Supercritical Fluid Chromatography TFA trifluoroacetic acid TGI tumor growth inhibition

[0363] Example 1: Synthesis of (R)-2-amino-5-(4-(2-(3,5-difluorophenyl)-2-hydroxyacetamido)-2-methylphenyl)-N-isopropylnicotinamide (compound of formula (I)) and hemifumarate salt Step 1: Isolation of (2R)-2-(3,5-difluorophenyl)-2-hydroxy-acetic acid [ka]

[0496] Prior to use, Lipase PS Amano (see Mendiola, J. et al., Org. Process Res. Dev. 2012, 16, 1312-1316) was immobilized on a diatomaceous earth support by mixing 200 g of diatomaceous earth with 200 g of Lipase PS Amano SD. H2O was added until the solid was covered, and the mixture was stirred. The H2O was removed in an oven at 4 mbar and 40°C for 16 hours. Water content was determined by Karl Fischer titration to be less than 1%.

[0364]

[0497] Support-immobilized lipase PS Amano SD (250 g) and vinyl acetate (312 mL; 3.36 mol) were added to a suspension of racemic 2-(3,5-difluorophenyl)-2-hydroxyacetic acid (125 g, 664 mmol) in methyl tert-butyl ether (2.50 L), and the mixture was stirred at 26° C. for 72 h. After this time, the solid was rinsed and filtered with methyl tert-butyl ether (1.50 L), and the combined filtrate was concentrated under reduced pressure. The residue was slurried in dichloromethane (160 mL) at 23° C. for 4 h. The mixture was filtered, and the solid was washed with petroleum ether (150 mL), and dried to give the title compound (47.0 g, 36%). 1H NMR (d6-DMSO) δ 5.11 (s, 1H), 6.20 (bs, 1H), 7.11-7.21 (m, 3H), 12.8 (bs, 1H). The absolute configuration was determined by vibrational circular dichroism (see Freedman TB et al, Chirality, 2003 Nov., 15(9), 743-758). Chiral HPLC: Rt = 7.39 min (UV); Column: Chiralpak® AD 4.6 × 150 mm 5 μm; 5% EtOH in n-hexane (0.05% TFA) isocratic; Flow rate: 1.5 mL / min, ee > 98%.

[0365] Step 2: Synthesis of 2-amino-5-(4-amino-2-methyl-phenyl)-N-isopropyl-pyridine-3-carboxamide [ka]

[0498] 3-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (93.6 g, 0.401 mol), K2CO3 (119 g, 0.860 mol), and Pd(dppf)2Cl2 (10.6 g, 140 mmol) were added to a solution of 2-amino-5-bromo-N-isopropyl-pyridine-3-carboxamide (74.0 g, 0.287 mol) in dioxane (888 mL) and HO (296 mL), and the mixture was heated overnight at 55° C. The mixture was cooled to 23° C., ethyl acetate (150 mL) was added, and the resulting suspension was filtered through diatomaceous earth, rinsing the solid with ethyl acetate (50 mL). The combined filtrate was washed with H2O (30 mL) and saturated aqueous NaCl (300 mL) and concentrated under reduced pressure to give the title compound (78.0 g, 96%). ES / MS m / z 285.1 (M+H).

[0366] Step 3: Synthesis of (R)-2-amino-5-(4-(2-(3,5-difluorophenyl)-2-hydroxyacetamido)-2-methylphenyl)-N-isopropylnicotinamide [ka]

[0499] A mixture of (2R)-2-(3,5-difluorophenyl)-2-hydroxyacetic acid (29.0 g, 0.154 mol), 2-amino-5-(4-amino-2-methyl-phenyl)-N-isopropyl-pyridine-3-carboxamide (43.83 g, 0.154 mol), and N,N-diisopropylethylamine (39.8 g, 0.308 mol) in tetrahydrofuran (960 mL) was treated with (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate) (87.9 g, 0.231 mol) at 0° C. for 30 minutes, and the mixture was warmed to 20° C. and stirred for 2 hours. Ethyl acetate (50 mL) was added, and the mixture was filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by chromatography (eluent: 2:1 petroleum ether / ethyl acetate) and then by supercritical fluid chromatography, SFC (column: Chiralpak® IC 30×250 mm 5 μm (Daicel); MeOH / CO2=30:70 isocratic; flow rate: 80 g / min; back pressure: 100 bar; column temperature: 40 °C) to give the title compound (27.5 g, 39%) as a white solid. ES / MS m / z 455.2 (M+H).

[0367] Step 4: Synthesis of (R)-2-amino-5-(4-(2-(3,5-difluorophenyl)-2-hydroxyacetamido)-2-methylphenyl)-N-isopropylnicotinamide hemifumarate [ka]

[0500] To a 300 L reactor under N2 and stirring at 25°C, IPA (174.3 kg, 40.0 V) and HC-5404 (5.48 kg, 0.012 mol, 1.0 eq) were added. The resulting mixture was stirred at 35°C for 3 hours and filtered through a 3M carbon filter (IPC: Pd content <2 ppm LOQ). The filtrate was combined with fumaric acid (0.72 kg, 0.55 eq) and stirred at 25°C for 9 hours to give HC-5404-FU.

[0368] Example 2: Study of HC-5404 + anti-PD-1 antibody (RMP1-14 murine antibody) combination therapy in a murine MB49 syngeneic bladder cancer model

[0501] C57BL / 6 mice were inoculated subcutaneously with MB49 cells, and treatment began 8 days after cell inoculation. Groups of animals (n = 10 / group) received either vehicle, HC-5404 (PO, BID) (3 mg / kg, 10 mg / kg, or 30 mg / kg), 5 mg / kg anti-PD-1 antibody (aPD-1) (RMP1-14) (IP, BIW), or a combination of both. Animals were sacrificed at various time points, and flow cytometry was performed on blood or single-cell suspensions from tumors or lymph nodes (n = 6). Although HC-5404 alone showed only a modest antitumor effect (32% tumor growth inhibition (TGI)), adding HC-5404 to aPD-1 resulted in a combined antitumor effect (75% TGI), significantly improving the effect of aPD-1 alone (53% TGI) (Figures 2B, 2C, 2D, 2E, 2F, and 2G).

[0369]

[0502] The efficacy of HC-5404 + aPD-1 therapy correlated with increased expression of type 1 interferon receptor (IFNAR1) and surface calreticulin on tumor cells (Figures 3D and 4B). In addition, IFNAR1 expression was significantly increased on polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) and tumor-associated macrophages (TAMs) (Figures 3A, 3B, and 3C). TAMs also showed increased expression of PD-L1 with combination treatment (Figure 3E). Concomitant with myeloid cell activation, combination treatment increased the frequency of CD8 T cells in tumors (Figures 5A and 5B), along with increased expression of the activation marker CD69 on T cells in tumor-draining lymph nodes (Figures 5C and 5D). Notably, the effect of HC-5404 on IFNAR1 was also detected on peripheral blood monocytes, demonstrating surface expression of IFNAR1 as a potential biomarker for HC-5404 activity (Figure 8). In vitro evaluation of human umbilical cord blood- and mouse bone marrow-derived MDSCs demonstrated reduced T cell inhibition in the presence of HC-5404 (Figures 10A, 10C, 11A, and 11C). Collectively, these data demonstrate the effective immunostimulatory effect of coadministering HC-5404 with anti-PD1 mAb and outline its potential application in cancers under ICI treatment.

[0370]

[0503] HC-5404 + aPD-1 treatment also correlated with increased CD4 activation and memory T cell induction (as evidenced by Ki67 expression, EOMES expression, granzyme B expression, TCF-1 expression, and / or effector memory populations) (Figures 6A, 6B, 6C, 6D, and 6E) and expansion of DC populations in the lymph nodes (LNs) of treated mice (Figures 7A, 7B, and 7C).

[0371] [Table 1]

[0372] Animal characterization

[0504] Female C57BL / 6 mice (7-8 week old animals, 20-25 grams, Envigo) were used.

[0373] cell line

[0505] The MB49 (Lot No. 18B016) murine urothelial carcinoma cell line was purchased from Sigma-The European Collection of Authenticated Cell Cultures (ECACC). The cells were free of human and animal pathogens when tested by h-IMPACT-I and IMPACT-III (IDEXX Bioresearch, Columbia, MO). In addition, the genetic profile of the cells was consistent with the ECACC reference standard when tested by STR-based DNA fingerprinting assay. The cells were maintained in DMEM containing 10% FBS and cultured up to passage 4 before transplantation. Cells were harvested at approximately 80% confluence, washed twice with DMEM without FBS, and cultured at 5 × 10 cells for transplantation. 6 The cells were suspended in FBS-free DMEM at 1000 cells / mL.

[0374] Building the MB49 model

[0506] On day 0, female Balb / c mice (7-8 weeks old) under isoflurane anesthesia were injected with 0.5 × 10 infusions in a total volume of 100 mL into the right flank area. 6 MB49 cells were subcutaneously implanted into each mouse. Five days after implantation, the average tumor volume was approximately 50 mm. 3 When the serotonin concentration reached 0.05, animals were randomized into treatment groups.

[0375] Measurement parameters

[0507] Tumor volumes were measured twice weekly.

[0376]

[0508] Tumor volume was calculated using the formula [tumor volume (mm 3 ) = π / 6 × length × width 2) and plotted as mean ± SEM. %T / C (ratio of tumor volume between treatment and control groups) was calculated by the formula 100 × ΔT / ΔC if the mean ΔT > 0. ΔT, mean tumor volume of the drug-treated group on the observation day of the study - mean tumor volume of the drug-treated group on the start of dose administration; ΔC, mean tumor volume of the control group on the observation day of the study - mean tumor volume of the control group on the start of dose administration. If ΔT < 0, regression was calculated using the formula = 100 × ΔT / T start. If tumor volume was 14 mm or less for three consecutive measurements, regression was calculated using the formula = 100 × ΔT / T start. 3 Animals with less than 50% tumor regression were considered complete responses, and tumors with greater than 50% regression were considered partial responses.

[0377]

[0509] At the end of the study, animals were euthanized using CO2 asphyxiation.

[0378] statistical analysis

[0510] Tumor volume data were analyzed statistically using two-way ANOVA followed by Dunnett's multiple comparison test (GraphPad Prism).

[0379] Flow cytometric analysis of myeloid and lymphoid cells in MB49 Preparation of tumor tissue

[0511] Tumor tissue samples were obtained 7 days after treatment (14 days after inoculation). Single cells were prepared by enzymatic digestion of tumor tissue and subsequently analyzed by flow cytometry on a Cytek Aurora. Expression of IFNAR1 (A) and PD-L1 (B) in myeloid populations and tumors is shown.

[0380]

[0512] Seven days after treatment, surface calreticulin on MB49 tumor cells was analyzed by flow cytometry.

[0381] LN preparation

[0513] Single-cell suspensions were washed twice with PBS and then stained with LIVE / DEAD fixable blue dead cell stain (Invitrogen catalog no. L23105) at a 1:3000 dilution for 30 minutes at 4°C. Cells were washed once with FACS buffer (PBS containing 2.5% FBS and 0.1% sodium azide), Fc receptors were blocked for 5–10 minutes, and then stained with an Ab cocktail in Super Bright complete staining buffer (Invitrogen catalog no. SB-4401) for 30 minutes at 4°C in the dark. Labeled cells were then fixed with fixation / permeabilization buffer (Tonbo Biosciences) for 30 minutes at 4°C, washed three times with permeabilization buffer (Tonbo Biosciences), and stained with intracellular Abs for 30 minutes at 4°C. Stained cells were then washed sequentially once with permeabilization buffer and twice with FACS buffer. Stained cells were analyzed by flow cytometry on a Cytek Aurora (Cytek Biosciences) platform, and data were analyzed by FlowJo (BD).

[0382]

[0514] LN cells from samples taken 14 days after treatment were analyzed by flow cytometry, showing expression of Ki67, EOMES, and granzyme B on CD4+ T cells, effector memory (EM) populations, and TCF-1 expression on activated T cells. Effector memory (EM) CD4+ cells express CD62L - CD44 + CD4 + All DCs are defined as CD11c + MHCII + CD11b dim / - and cDC1 was identified as XCR1 + DCs and cDC2s were identified as XCR1 - CD24 + Identify as DC.

[0383] peripheral blood cells

[0515] In the MB49 model, whole blood was collected on the indicated days after treatment. IFNAR1 expression was determined by flow cytometry, gating on CD14+ monocytes.

[0384] [Table 2]

[0385] [Table 3]

[0386] [Table 4]

[0387] HMGB1 IHC staining

[0516] HC-5404 has demonstrated efficacy as a single agent and in combination in multiple solid tumor xenograft models.

[0388]

[0517] 786-O tumor cells were maintained in vitro at 37°C in a 5% CO atmosphere using MEM medium supplemented with 10% heat-inactivated fetal bovine serum and 0.01 mM NEAA. Each mouse was inoculated subcutaneously in the right front with 786-O tumor cells (5 × 10) in 0.1 ml of PBS (1:1 Matrigel) and allowed to develop tumors. The average tumor size was approximately 200–300 mm. 3 Randomization for efficacy studies begins when the 3 It started with.

[0389]

[0518] Animals were treated with HC-5404 at 30 mpk BID for 28 days. Animals were euthanized and tumors were harvested, fixed in 10% neutral buffered formalin, and paraffin-embedded for IHC analysis.

[0390]

[0519] Paraffin sections of tumor samples were stained for HMGB1 on slides using Abcam antibody (catalog no. ab18256) and chromogenic DAB stain in Bond Rx (Leica Biosystems). Slides were digitized using a Leica AT2 slide scanner with a 40x objective. Scanned images were analyzed using a custom-written macro in ImageJ / FIJI (NIH) to measure nuclear and non-nuclear signals (Figures 4C and 4D).

[0391] RNA purification

[0520] Mouse tumor, spleen, and lung tissues were rapidly frozen and stored in a -800°C freezer. The frozen tissues were then immersed in 10 volumes of RNAlater®-ICE (ThermoFisher) at -200°C for over 16 hours. The tissues were then transferred to M tubes (Miltenyi Biotech) containing RLT lysis buffer (Qiagen) and homogenized using a gentleMACS tissue dissociator under program RNA_02 (Miltenyi Biotech). Total RNA was isolated from the homogenized tissues using the RNeasy Plus Mini Kit (Qiagen). RNA concentrations were determined using an Epoch microplate spectrophotometer (Biotek).

[0392] QuantiGene® Plex Assay

[0521] A custom-made 77-plex Mouse Immune QuantiGene Assay Kit (ThermoFisher Scientific) using a bead-based multiplex assay was used to measure the expression of 69 immune genes, 4 stress genes + 4 reference genes on a Luminex xMAP 200 (ThermoFisher Scientific) according to the manufacturer's instructions.

[0393] Data analysis

[0522] The detection limit for each gene analyzed was determined by subtracting the mean intensity of the negative control wells from each unknown sample. The target gene expression values ​​were normalized to the geometric mean of four housekeeping genes. The fold change for each treatment group relative to the mean control was log2 transformed and presented as a heatmap (Figure 9).

[0394]

[0523] Data were analyzed using Prism software (version 9.1.0) using one-way ANOVA with post hoc (Tukey) multiple comparison tests and are expressed as mean ± SEM. A p value of less than 0.05 was considered statistically significant.

[0395] Human and mouse MDSC research Human MDSCs reagent Lymphoprep (StemCell Tech Catalog No. 07851) EasySep Human Umbilical Cord Blood CD34 Positive Selection Kit II (StemCell Tech Catalog No. 17896) IMDM medium (Gibco catalog number 12440-046) RPMI (Corning catalog number 10-040-CV) FBS (Gibco catalog number 10438-026) Penicillin-streptomycin (Gibco catalog number 15140122) L-glutamax (Gibco catalog number 35050-061) Human GM-CSF (R&D systems catalog number 215-GM) Human G-CSF (R&D systems catalog number 214-CS) Human SCF (R&D systems catalog number 7466-SC) EasySep Human T Cell Isolation Kit (StemCell Tech Catalog No. 17951) Dynabeads Human T-activator CD3 / CD28 (Gibco Catalog No. 11131D) Cell Trace Violet (CTV) Cell Proliferation Kit (Invitrogen Catalog No. C34557) IL-2 (R&D systems catalog number 202-IL)

[0396]

[0524] Human MDSC preparation: Fresh umbilical cord blood was obtained from the NDRI (Protocol ID No. RCHA4 01). Mononuclear cells were enriched from the cord blood by Lymphoprep density gradient centrifugation, followed by isolation of CD34+ cells using the EasySep Human Umbilical Cord Blood CD34 Positive Selection Kit II. The enriched CD34+ cells were cultured in 48-well plates (BD Falcon, Bedford, MA) at 1 x 105 cells / well in 1 ml of IMDM complete medium (IMDM supplemented with 10% FBS, 1x penicillin-streptomycin, and 2 mM L-glutamax). IMDM complete medium was supplemented with 100 ng / ml human GM-CSF, 100 ng / ml human G-CSF, and 50 ng / ml human SCF. The cells were expanded at 37°C in a 5% CO2 humidified incubator for 13–14 days, with fresh medium replaced on day 7. Expand CD34+ cells to 0.1–0.15 × 10 in 24-well plates. 6 MDSC differentiation was induced by culturing cells / well in 2 ml of IMDM complete medium supplemented with 100 ng / ml human GM-CSF, 50 ng / ml G-CSF, and 30 ng / ml IL-6 for an additional 6–7 days, with a fresh medium change on day 3. For HC-5404 treatment, the indicated dose of HC-5404 was added to the cell culture medium on day 0 of the MDSC differentiation phase, and the medium was changed to fresh medium on day 3, with additional HC-5404 added. MDSCs were harvested on day 6 or 7 and used in T cell suppression assays.

[0397]

[0525] Human T cell suppression assay. PBMCs were enriched by Ficoll density gradient centrifugation from Trima Cones obtained from Innovative Blood Resources (Protocol ID number 120160627). CD3 T cells were further enriched from PBMCs using the EasySep Human T Cell Isolation Kit. CD3 T cells were labeled with Cell Trace Violet (CTV) according to the supplier's protocol. CTV-labeled T cells were resuspended in RPMI medium supplemented with 10% FBS, 1x penicillin-streptomycin, and 2 mM L-glutamax and added to a 96-well round-bottom plate at 50,000 cells per well. Dynabeads human T-activator CD3 / CD28 were washed twice with PBS and then added to the T cells at a 1:1 ratio. HC-5404-treated and untreated MDSCs were harvested from the plate, washed three times with RPMI medium, and then added to wells at various MDSC:T ratios (4:1, 2:1, or 1:1). IL-2 (30 μg / ml) was added to each well, and the 96-well plates were incubated at 37°C in a 5% CO2 humidified incubator for 3–5 days. T cell proliferation was analyzed by flow cytometry. Cells were washed two to three times with PBS and then stained with a fluorescently conjugated anti-CD3 antibody for 30 minutes at 4°C in the dark. Labeled cells were analyzed using a BD Fortessa cytometer, and data were analyzed using FlowJo. Statistical analysis was performed using Prism software (GraphPad Software). Data were analyzed using one-way ANOVA with multiple comparisons corrected by Dunnett's test. * P < 0.05; ** P < 0.01; *** P<0.005, **** P<0.0001 (Figures 10A, 10B, and 10C).

[0398] Murine MDSCs reagent RPMI (Corning catalog number 10-040-CV) FBS (Gibco catalog number 10438-026) Penicillin-streptomycin (Gibco L-glutamax (Gibco catalog number 35050-061) HEPES (Gibco catalog number 15630-080) Sodium pyruvate (Gibco catalog number 11360-070) 2-ME (Gibco catalog number 21985-023) MEM NEAA (Gibco catalog number 11140-050) RPMI without glucose (Gibco catalog number 118790-020) Glucose (Gibco Catalog No. A24940-01) Mouse GMCSF (PeproTech Catalog No. 315-03) Mouse IL-6 (PeproTech Catalog No. 216-16) EasySep Mouse T Cell Isolation Kit (StemCell Tech Catalog No. 19851A) Dynabeads Mouse T-activator CD3 / CD28 (Gibco Catalog No. 114543D)

[0399]

[0526] Murine MDSC preparation: Bone marrow-derived single cell suspensions were prepared from the tibia and femur of C57BL / 6 mice. 2 2.5 × 10 cells per 10 ml in RPMI complete medium (RPMI medium supplemented with 10% FBS, 10 mM HEPES, 1 mM sodium pyruvate, 50 μM 2-ME, 1 mM MEM NEAA, 1× penicillin-streptomycin, and 2 mM L-glutamax) in a tissue culture dish. 6Cells were cultured at 37.0°C in a 5% CO2 humidified incubator for 4 days. On day 0, cytokines GM-CSF (10 ng / ml) and IL-6 (10 ng / ml) were added to the cell culture medium. For low-serum, low-glucose (LSLG) medium treatment, the cultured cells were first harvested from the plate and washed twice with RPMI medium to remove residual serum and glucose. They were then cultured for 6 hours in 5 ml of LSLG medium (glucose-free RPMI supplemented with 1% FBS, 1 g / L glucose, 1x penicillin-streptomycin, and 2 mM L-glutamax) in a 15 ml conical tube with or without HC-5404. Cells were harvested and used for T cell suppression assays.

[0400]

[0527] Mouse T cell suppression assay: Spleens were harvested from C57BL / 6 mice. Single-cell suspensions were prepared, and CD3 T cells were isolated using the EasySep Mouse T Cell Isolation Kit. CD3 T cells were labeled with Cell Trace Violet (CTV) according to the supplier's protocol. CTV-labeled T cells were resuspended in RPMI medium supplemented with 10% FBS, 1x penicillin-streptomycin, and 2 mM L-glutamax and added to a 96-well round-bottom plate at 100,000 cells per well. Dynabeads mouse T-activator CD3 / CD28 were washed twice with PBS and then added to the T cells at a 1:1 ratio. LSLG ± HC5404-treated and untreated MDSCs were harvested from the plate, washed three times with RPMI medium, and then added to the wells at various ratios of MDSC:T (8:1, 4:1, 2:1). IL-2 (30 μg / ml) was added to each well, and the 96-well plate was incubated at 37°C in a 5% CO2 humidified incubator for 3–5 days. T cell proliferation was analyzed by flow cytometry. Cells were washed 2–3 times with PBS and then stained with fluorescently conjugated anti-CD4 and anti-CD8 antibodies for 30 minutes at 4°C in the dark. Labeled cells were analyzed on a BD Fortessa cytometer, and data were analyzed using FlowJo. Statistical analysis was performed using Prism software (GraphPad Software). Data were analyzed using one-way ANOVA with multiple comparisons corrected by Dunnett's test. * P < 0.05; ** P < 0.01; *** P<0.005, **** P<0.0001 (Figures 11A, 11B, and 11C).

[0401] Example 3: Cell-based assay

[0528] Cell-based assay: 1 x 10 HEK293 cells per well 6Cells were plated and treated with HC-5404 for 30 minutes, after which tunicamycin (1 μM) was added to induce ER stress. Four hours after tunicamycin addition, cells were harvested and lysed in RIPA buffer supplemented with protease and phosphatase inhibitors for Western blotting. Total protein concentration was determined by BCA, and 30 mg of protein was loaded onto an SDS-PAGE gel, transferred to nitrocellulose, and probed with primary antibodies against phosphorylated T-982 of human PERK or total ATF4, followed by incubation with an infrared-labeled secondary antibody. Blots were imaged on a LiCor Imager and normalized to the total protein in the well using Revert™ staining. Band intensities were quantified and normalized to those of DMSO-treated samples. Normalized band intensities were plotted as percent inhibition against a 10-point, 3-fold dilution series of HC-5404. IC values ​​were calculated using a four-parameter logistic curve fit in XLFit. 50 values ​​were calculated.

[0402] Example 4: Biochemical Assays

[0529] The potency of HC-5404 against the four ISR kinases was assessed in a cell-free FRET-based biochemical assay according to the manufacturer's instructions (Carna).

[0403] Example 5: Protein Analysis

[0530] Samples were placed in 500 μL of Novex lysis buffer, consisting of 2x Novex Tris-Glycine SDS sample buffer (ThermoFisher LC2676), 1x Phosphatase Inhibitor Cocktail III (Sigma P0044), 1x Phosphatase Arrest I (Gibco 113P-D), 1x 2-mercaptoethanol (Gibco 21985023), 1x Benzonase (EMD Millipore Corp 70746-3), and 1x Protease Inhibitor Cocktail Tablet (Roche / Sigma 11836170001). The samples were homogenized with a Polytron and heated at 95°C for 10 minutes. They were then centrifuged at maximum speed for 10 minutes, and the supernatant was decanted. The protein concentration of the samples was measured using the Pierce 660nm Protein Assay Kit (Thermo Scientific 22660) according to the manufacturer's instructions.

[0404]

[0531] Western blot analysis was performed on a SimpleWestern high-throughput protein analysis platform (ProteinSimple 004-650) using the 12–230 kDA separation module (ProteinSimple SM-W004) and the total protein detection module (ProteinSimple DM-TP01) according to the manufacturer's protocol. The antibodies used were: pPERK (Lilly; 1:50) and PERK (Cell Signaling Technology 5683; 1:200) (additional information on other antibodies used). All protein analytes were normalized to the total protein loaded onto the capillary.

[0405] Example 6: Immunohistochemistry and image analysis

[0532] Formalin-fixed, paraffin-embedded samples were sectioned at 5 μm thickness and mounted on Superfrost Plus microscope slides (Fisher Scientific). All immunohistochemistry (IHC) staining was performed on a Bond Rx automated stainer (Leica Biosystems) using the conventional TSA amplification and detection system. The primary antibodies used were: α-CD31 (Abcam #ab182981), α-smooth muscle actin (SMA; Abcam #ab124964), and α-Meca32 (Novus Biologicals #NB100-77668). TSA-conjugated Alexa488 and Alexa647 fluorophores (Invitrogen #B40953 and #B40958) were used at a 1:500 dilution. Slides were counterstained with DAPI and coverslipped using Mowiol antifade mounting medium (Sigma #D2522).

[0406]

[0533] Slides were imaged using a 10x / 0.32NA objective on an Aperio Versa 200 (Leica Biosystems) whole-slide scanner, and data were analyzed using a custom-written macro in ImageJ / FIJI (NIH). Briefly, a 150 μm outer edge of tumor sections was excluded from analysis. Tissue area was measured using the DAPI channel with Gaussian blurring. Scans of 104envat staining were processed with a median filter to remove noise, and then the positive area was measured at the appropriate threshold level. The percent positive area for each marker was calculated. CD31-SMA double-positive area was measured using image calculations on binary images. These data were subjected to one-way ANOVA and pairwise comparisons using Prism GraphPad.

[0407] Example 7: In vivo mouse tumor studies animal

[0534] Animals were allowed to acclimate for 7 days after arrival in the animal facility. Mice were housed under pathogen-free conditions in a facility with a 12-hour light / dark cycle and were provided with standard laboratory chow and water ad libitum.

[0408] Xenograft model

[0535] The xenograft RCC model was established as follows: female NOD.SCID mice (6-8 weeks old) were inoculated with A498 RCC cells (5 × 10 6 Female Balb / c nude mice (6–8 weeks old) were injected with 786-O RCC cells (5 × 10 6 cells / mouse) or Caki-1 RCC cells (6 × 10 6 cells / mouse) and MFE280 endometrial cells (10 × 10 6 The cells were transplanted into the flank of the animals.

[0409] PDX Model

[0536] To generate patient-derived xenograft models, small pieces of neuroendocrine pancreatic carcinoma were implanted into female NSG mice (6–8 weeks old), fragments of Huprime gastric carcinoma were implanted into female Balb / c nude mice, and for single-animal studies in RCC PDX models, tumor fragments from selected RCC PDX models were implanted into one side of mice.

[0410] treatment

[0537] Tumors typically 150-250 mm 3 Once tumor volume reached 100 μg, animals were randomized by tumor volume into treatment groups with 8 or 10 animals per group. Single-animal studies in RCC PDX models enrolled only one animal per group. Treatment groups typically included vehicle (10 μl / g body weight, orally (PO), twice daily (BID)), HC-5404 (3, 10, or 30 mg / kg PO, BID), cabozantinib (30 mg / kg PO, daily (QD)), axitinib (30 mg / kg PO, BID), envatinib (10 mg / kg PO, BID), sunitinib (20 mg / kg PO, QD), or DC101 (15 mg / kg, 15 mg / kg, or 20 mg / kg, intraperitoneally (IP), twice weekly (BIW)).

[0411]

[0538] Body weight and tumor volume were measured twice a week. 3 ) = π / 6 × length × width2 Tumor volumes were calculated using the % ΔT metric and plotted as the mean + SEM. At the end of the study, if ΔT > 0, the percentage of tumor growth inhibition was calculated using the formula: 100 - [1 - (ΔT / ΔC)] (ΔT, mean tumor volume of the drug-treated group on the observation day of the study - mean tumor volume of the drug-treated group on the first dose; ΔC, mean tumor volume of the control group on the observation day of the study - mean tumor volume of the control group on the first dose). If ΔT < 0, regression was calculated using the formula = 100 × ΔT / T at start. Animals with greater than 50% regression were considered partial responders. Animals with a tumor volume of 0 (no measurable tumor) were considered complete responders, and animals with less than 50% tumor regression and less than 30% growth compared to baseline were considered stable disease. Animals with greater than 30% tumor growth compared to baseline were considered progressive disease. The percent body weight change was calculated using the formula (body weight on observation day - body weight on day 1) / body weight on day 1 × 100%.

[0412] Multiple in vivo studies

[0539] At the start of the study, 786-O xenografts (approximately 200 mm 3 One hundred and sixteen mice bearing HC-5404 (30 mg / kg PO; BID) initiated treatment with axitinib as a single agent (30 mg / kg PO; BID). After 14 days of treatment, 77 mice bearing the largest tumors were re-randomized into four PD arms (n=8) and four efficacy arms (n=10). Five baseline tumor samples were collected on the day of re-randomization. The remaining animals were removed from the study. The four treatment groups were vehicle, HC-5404 (30 mg / kg PO; BID), axitinib (30 mg / kg PO; BID), or a combination thereof, and these treatments continued for 28 days.

[0413] Example 8: HC-5404 is a potent and selective PERK inhibitor

[0540] PERK is one of four closely related kinases of the integrated stress response (ISR) that phosphorylates eIF2α in response to cellular stress. Both tumorigenesis and treatment with many anticancer drugs have been shown to activate the ISR, and hypoxic stress is associated with activation of PERK signaling as an adaptive response. To understand the specific role of PERK in RCC, we investigated the aminopyridine PERK inhibitor HC-5404.

[0414]

[0541] The selectivity of HC-5404 for PERK relative to the ISR kinases GCN2, HRI, and PKR was assessed using a cell-free biochemical assay. In this assay, recombinant target kinases were incubated with ATP and an eIF2α-based fluorogenic substrate, and kinase activity was assessed in the presence of HC-5404 over a concentration series. These experiments demonstrated the IC 50 The activity of 1 nM was found to be greater than 2000-fold biochemically selective over GCN2 (IC50 = 2.17 μM), HRI (IC50 = 2.96 μM), and PKR (IC50 > 10 μM) (Figures 12, 13, 14, 15, 16).

[0415]

[0542] Next, we evaluated the selectivity of HC-5404 across a broader kinome by measuring binding to 468 unique kinases, including 403 wild-type human kinases and 59 mutant isoforms, using the KINOMEScan biochemical panel assay. Of note, this kinome panel does not include PERK (also known as EIF2AK3). HC-5404 was evaluated at 100 nM, 1000 nM, and 10,000 nM, revealing a high degree of selectivity across this kinome (Figure 17). No interactions were observed when HC-5404 was assayed at 100 nM, whereas BLK and the atypical kinase RIOK2 were inhibited at 1000 nM, and only nine kinases were inhibited by >65% at 10,000 nM HC-5404.

[0416]

[0543] The cell-based activity of HC-5404 was evaluated using HEK293 cells treated with tunicamycin (Tm; 1 μM) to induce ER stress. Tm treatment resulted in autophosphorylation of PERK at Thr982 (pPERK) and increased ATF4 levels, both of which were inhibited by HC-5404 in a concentration-dependent manner (IC ). 50 =23 nM and 115 nM; Figures 12, 13, 14, 15, 16).

[0417]

[0544] Taken together, these results demonstrate that HC-5404 is a potent, selective and cell-active PERK kinase inhibitor.

[0418] Example 9: In vivo characterization of HC-5404

[0545] Initial in vivo studies were conducted to understand pharmacokinetic and pharmacodynamic (PK / PD) relationships and optimize dose administration regimens. HC-5404 was quantified in mouse plasma after single oral administration (PO) at doses ranging from 3 to 100 mg / kg. Up to 100 mg / kg, a dose-proportional increase in exposure was observed, with Cmax occurring before 1 hour and a mean half-life of 2.22 hours. Mouse plasma protein binding (PPB%) was determined to be 95.6%, allowing us to calculate free drug plasma exposure over time (Figure 18). A 30 mg / kg oral dose resulted in a free drug exposure of 355 nM at Cmax, well below the 1000 nM concentration assayed by kinome scan, which revealed minimal secondary binding.

[0419]

[0546] The PK / PD relationship was evaluated in mouse pancreas, as the pancreas exhibits elevated pPERK activity and is therefore suitable for PD analysis. Treatment with HC-5404 resulted in time- and dose-dependent inhibition of p-PERK in pancreatic tissue. When administered at 30 mg / kg, plasma HC-5404 reached 3668 ng / mL (355 nM, unbound) at 1 hour, as shown in the table below. At these concentrations, pPERK inhibition was approximately 90% (Figure 19). As HC-5404 was cleared from plasma, pPERK gradually increased and returned to basal levels by 12 hours after administration.

[0420] [Table 5]

[0421]

[0547] A single oral dose of 100 mg / kg achieved a proportional increase in exposure, reaching a Cmax of 16583 ng / mL (1605 nM, unbound) (Figure 20). At this high dose level, HC-5404 exposure remained above 3200 ng / mL (311 nM, unbound) in plasma for up to 12 hours after administration, resulting in pPERK inhibition (<20% activity) for the first 12 hours after treatment.

[0422]

[0548] The ability of HC-5404 to suppress tumor growth was evaluated in the 786-O xenograft RCC model. The 786-O tumor model is homozygous for a frameshift mutation following Gly104 in the von Hippel-Lindau protein (VHLmut), resulting in activation of the UPR, including HIF stabilization and elevated pPERK expression. Similar to the pancreas, administration of 30 mg / kg HC-5404 induced approximately 90% pPERK inhibition in 786-O tumors 1 hour after dose administration, which returned to baseline by 8 hours (Figure 21).

[0423]

[0549] To evaluate the single-agent activity of HC-5404 and explore the effect of splitting the dose between QD and BID administration, mice bearing 786-O xenografts were treated with 3, 10, or 30 mg / kg PO BID, in parallel with 6, 20, or 60 mg / kg PO QD, to assess the effect of the dosing regimen on tumor growth. Although the difference between BID and QD administration was not statistically significant, the general trend indicated a better antitumor effect with BID administration. Of the doses tested, 30 mg / kg BID induced the highest efficacy, with 48% tumor growth inhibition (Figure 22). Based on these results, the 30 mg / kg BID dose was used for subsequent in vivo tumor studies.

[0424]

[0550] The dose level of HC-5404 was further explored using Capan-2 pancreatic tumor xenografts, a model previously reported to be sensitive to PERK inhibition. Treatment with 30 and 100 mg / kg HC-5404 BID resulted in 50% and 43% TGI, respectively (Figure 23). The lack of differences between dose levels suggests that increasing the dose above 30 mg / kg when administered twice daily is unlikely to improve the single-agent activity of HC-5404. Because PERK inhibition has been associated with pancreatic toxicity, pancreatic sections from mice treated with 30 mg / kg or 100 mg / kg HC-5404 were evaluated. Only the 100 mg / kg dose revealed histological changes in the pancreas, which were reversible with a 2-week washout period (Figure 24). Taken together, these findings further supported the decision to administer a dose of 30 mg / kg HC-5404 in subsequent in vivo studies.

[0425] Example 10: HC-5404 enhances the sensitivity of RCC tumor models to VEGFR-TKIs

[0551] Receptor tyrosine kinase inhibitors (RTKs) targeting VEGF receptors (VEGFR-TKIs) are antiangiogenic agents that disrupt tumor vascular development as a critical component of their mechanism of action, driving tumor hypoxia and nutrient deprivation. Hypoxia and glucose deprivation are known drivers of ER stress and PERK pathway activation in tumors.

[0426]

[0552] The effects of three second-generation VEGFR-TKIs on tumor growth and PERK activation were evaluated. Mice bearing 786-O xenografts were orally administered multiple dose levels of cabozantinib (15, 30, and 60 mg / kg), lenvatinib (5 and 10 mg / kg), or axitinib (15 and 30 mg / kg) for 21 days, after which tumors were excised and analyzed for pPERK levels. All three VEGFR-TKIs induced a dose-dependent increase in pPERK, reflecting the level of tumor growth inhibition (Figure 26). The doses used were well tolerated, as no treatment-related weight loss was observed (Figure 26). Sunitinib was evaluated in 786-O xenografts and also demonstrated an increase in pPERK. This effect increased over time (Figure 27). Based on these experiments, the dose regimen for the follow-up combination study was selected.

[0427]

[0553] Next, studies were conducted to evaluate the effect of adding HC-5404. Axitinib, cabozantinib, lenvatinib, and sunitinib were each tested in combination with HC-5404 in studies using 786-O RCC xenografts. Each study included a PD arm, in which samples were collected 7 days after administration to evaluate the effect of treatment on angiogenesis and pPERK levels. Consistent with the initial dose-finding studies, the PD arms of these experiments revealed that all VEGFR-TKIs induced pPERK accumulation, which was inhibited by the addition of HC-5404 (Figure 27).

[0428]

[0554] In the efficacy arms of each tumor growth study, HC-5404 and second-generation VEGFR-TKIs (axitinib, lenvatinib, and cabozantinib) had a modest effect on tumor volume. In contrast, combining a VEGFR-TKI with HC-5404 resulted in tumor regression compared to baseline (Figure 28). At the dose levels tested, sunitinib was a more effective inhibitor of 786-O tumors compared with second-generation VEGFR-TKIs. While sunitinib as a single agent was an effective inhibitor of 786-O growth, TGI levels were still elevated in combination with HC-5404, supporting a mechanistic interaction between VEGFR-TKIs and PERK inhibitors and highlighting the potential therapeutic benefit of combining a VEGFR-TKI with HC-5404. HC-5404 sensitizes RCC tumor models to antiangiogenic VEGFR-TKIs and enhances their antitumor effects.

[0429]

[0555] The efficacy of HC-5404 was further validated in tumors by assessing the response of in vivo PERK inhibitory markers. Previous studies have highlighted the unexpected induction of downstream targets of ATF4, including asparagine synthetase (ASNS) and phosphoserine aminotransferase (PSAT1). Other canonical targets of ATF4, including CBS and CTH, were investigated. Treatment with HC-5404 induced dose-dependent accumulation of ASNS, CBS, and CTH in 786-O tumors (Figure 29).

[0430] Example 11: Sensitivity to the combination of VEGFR-TKI and HC-5404 is independent of VHL mutation status

[0556] RCC is characterized by frequent VHL mutations and their subsequent genetic silencing, which dysregulates HIF expression, driving a proangiogenic tumor environment associated with increased VEGF expression and activation of the UPR. There may be a correlation between VHL mutation status, pPERK expression, and sensitivity to VEGFR-TKIs. Three RCC tumor models were investigated: 786-O and A498 (Gly144 frameshift) were selected as representative VHL mutant models, and the Caki-1 model was selected as a VHL wild-type model.

[0431]

[0557] We first characterized the models by determining basal pPERK levels. Consistent with HIF pathway activation caused by VHL mutation-driven ER stress, 786-O and A498 tumors in the vehicle group exhibited elevated pPERK levels (Figure 30). In contrast, pPERK was nearly undetectable in Caki-1 tumors. As expected, treatment with HC-5404 potently inhibited pPERK expression in all three models (Figure 30).

[0432]

[0558] Because basal PERK pathway involvement may predict sensitivity to PERK inhibition, we examined the sensitivity of three RCC models to HC-5404. Sensitivity to HC-5404 varied among the three xenograft models, reflecting their basal levels of pPERK activity and VHL status. HC-5404 induced approximately 52% and 41% TGI in 786-O and A498, respectively, whereas HC-5404 had only minimal effects on growth in the Caki-1 model (Figure 36).

[0433]

[0559] To assess differences in sensitivity to PERK inhibition and VEGFR-TKI treatment, the A498 and Caki-1 models were treated with HC-5404, sunitinib, or their combination for 28 days. The level of tumor response among these three models reflected their genetic background: VHL mutant A498 was sensitive to HC-5404 as a single agent, but only achieved a TGI of approximately 40% (Figure 31). Similarly, the A498 model was sensitive to sunitinib, with robust single-agent activity resulting in a TGI of 80%. Combining sunitinib and HC-5404 in the VHL mutant model resulted in a 22% tumor regression in A498 (Figure 31). In contrast, Caki-1 was much less sensitive to either HC-5404 or sunitinib as monotherapy, and although the combination treatment was effective, the TGI only reached 47% in the combination group (Figure 32).

[0434]

[0560] The observation that Caki1 responded poorly to HC-5404 and sunitinib treatment suggested that VHL mutation status could potentially be used as a selection criterion to predict sensitivity to treatment. Therefore, 15 different RCC patient-derived xenografts (PDXs) were evaluated in a single-animal study directly comparing wild-type and mutant VHL models. Animals were treated with axitinib, HC-5404, or their combination for 28 days, and the magnitude of the observed combination benefit was scored. All five VHLwt models included in this study responded to the combination treatment, demonstrating significant improvement compared with monotherapy (representative growth curves are shown in Figures 33 and 34). Models were ranked based on tumor volume change compared to the starting baseline and scored for the number of partial responses (PRs). In this case, a PR was considered a volume regression of more than 50% compared to the starting baseline. Axitinib as monotherapy resulted in 1 / 15 PRs, whereas the combination of HC-5404 and axitinib resulted in 7 / 15 PRs (Figure 35). Importantly, while the VHLwt model retained its responsiveness to the combination treatment, three of these models appeared to have reduced sensitivity to axitinib. From this study, we conclude that HC-5404 improves tumor response to VEGFR-TKIs in various tumor contexts, and the effect is independent of VHL mutation status.

[0435] Example 12: HC-5404 demonstrates combination benefit with other agents that target angiogenesis without inducing tumor pPERK

[0561] Although a common target associated with antiangiogenic activity is inhibition of the VEGF axis, secondary targeting of VEGFR-TKIs may improve antitumor activity. To test whether inhibition of the VEGF / VEGFR axis was sufficient to drive the combination benefit with HC-5404 or whether a secondary effect was necessary, a murine monoclonal antibody selectively targeting VEGFR-2 was examined.

[0436]

[0562] DC-101 is a surrogate mAb for the human anti-VEGFR2 mAb ramucirumab and is approved for use in multiple cancer types. To determine whether treatment with HC-5404 could enhance the activity of DC-101, we tested DC-101 in three RCC models (786-O, A498, and Caki-1). Similar to observations with HC-5404 and multikinase-targeted TKIs, combination treatment significantly improved the efficacy of each monotherapy in 786-O and A498 xenografts, with TGI values ​​of 82% and 78%, respectively (Figure 37). In contrast, cotreatment of DC-101 with HC-5404 did not result in a significant combination benefit after 28 days of treatment in Caki-1, resulting in a TGI value of only 24% (Figure 37). Combination treatment did not result in tumor regression in any of the models tested.

[0437]

[0563] The effect of DC-101 on PERK activation was assessed in tumor samples excised after 4 days of treatment, and pPERK levels were determined by analyzing protein lysates. DC-101 did not affect pPERK levels in any of the three RCC models tested (Figure 37). In the 786-O and A498 models, pPERK was unchanged compared to the vehicle group, but HC-5404 inhibited pPERK signaling, as expected. In Caki1 samples, pPERK varied due to the low signal-to-noise data and was not affected by either DC-101 or HC-5404 treatment.

[0438] Example 13: PERK enhances the antiangiogenic effect of VEGFR-TKI

[0564] PERK is an adaptive stress response that links hypoxia and nutrient deprivation to proangiogenic signaling. We investigated the effect of HC-5404 on tumor vasculature when administered in combination with an antiangiogenic VEGFR-TKI. IHC staining of tumor cross sections confirmed that axitinib reduced the proportion of endothelial cells expressing Meca32 (also known as PLVAP), and the addition of HC-5404 enhanced this effect (Figures 38 and 39). Notably, HC-5404 alone had no effect on this endothelial marker, highlighting the context-dependent effects of HC-5404 on tumor angiogenesis.

[0439]

[0565] As immature vessels mature, they develop a layer of mural cells expressing smooth muscle actin (SMA), which surround and stabilize the maturing vessels. To determine whether treatment affects mature or immature tumor vessels, we performed co-staining for both CD31 and SMA. Similar to the effect observed with the endothelial marker Meca32, axitinib as a single agent reduced the proportion of immature CD31+SMA- vessels, while the combination treatment group enhanced it (Figure 39). In contrast, axitinib did not affect the number of mature CD31+SMA+ cells at the doses and time points analyzed, consistent with the central role of the VEGF axis in the generation of new vasculature through angiogenic sprouting. HC-5404 as a single agent did not affect the expression of either vascular marker, while combination treatment significantly reduced mature CD31+SMA+ vascular cells (Figure 39).

[0440] Example 14: PERK is a vulnerability of RCC xenografts that progressed during VEGFR-TKI treatment

[0566] Even if patients initially respond to VEGFR-TKI therapy, tumor regrowth necessitates subsequent rounds of therapy to improve patient outcomes. To model this effect, we challenged 786-O xenografts to progress during axitinib treatment and then added HC-5404 to axitinib treatment to see if the combination benefit could be maintained in this setting.

[0441]

[0567] The presence of a VEGFR-TKI during growth could drive increased sensitivity to PERK inhibitors. Tumors transitioning to the vehicle group showed an increased growth rate, suggesting that tumors had progressed during axitinib treatment but retained some sensitivity to treatment. Patients transitioning to single-agent axitinib or HC-5404 progressed at approximately the same rate as before re-randomization. By the end of the study, the single-agent activity of HC-5404 or axitinib inhibited tumor growth by 47% or 38% compared to vehicle (Figure 40). When HC-5404 was added to the axitinib regimen to create a combination group, xenografts continued to grow for a short period of time before shrinking in volume, resulting in a mean tumor regression of 20% compared to baseline (Figure 40). Thus, tumors that had already progressed during axitinib treatment retained sensitivity to HC-5404 when administered in combination with a VEGFR-TKI.

[0442]

[0568] Analysis of tumor vasculature revealed that both immature and mature blood vessels, including pericytes, were inhibited by the combination treatment. Consistent with previous findings, axitinib reduced the number of Meca32+ and CD31+ cells in xenograft sections, which was enhanced by the addition of HC-5404 in the combination group (Figures 41 and 42).

[0443]

[0569] Pericytes form the mural cell layer surrounding blood vessels and are important for vascular morphogenesis and branching. The effects of axitinib and HC-5404 on this layer were assessed using two pericyte markers (NG2 and MCAM). The proportion of NG2+ positively stained cells decreased only in the combination group compared to vehicle, highlighting the impact of the combination on this pericyte marker. MCAM+ cells were reduced by axitinib, an effect enhanced by the addition of HC-5404 (Figures 43 and 44). In both cases, the proportion of NG2+ and MCAM+ cells decreased relative to baseline samples, suggesting that, after re-randomization, tumors in the combination group experienced a loss of mature vascular cells rather than simply an inhibition of neovascularization. The combination of HC-5404 and axitinib inhibited tumor vasculature endothelial and pericyte markers.

[0444] Example 15: Adaptive stress drives resistance to VEGFR inhibitors

[0570] Antiangiogenic therapy increases tumor hypoxia, and tumor cells adapt to hypoxia by activating adaptive stress pathways, particularly PERK, which leads to cell survival.

[0445]

[0571] Following similar procedures as described in the above examples, 786-O RCC tumor xenografts were treated with cabozantinib (15, 30, 45 mg / kg; PO; QD), lenvatinib (5 and 10 mg / kg; PO; QD), and axitinib (15 and 30 mg / kg; PO; BID) for 21 days. VEGFR-TKIs were characterized in vivo and presented as tumor growth curves and pPERK / PERK levels, as shown in Figures 45, 46, and 47.

[0446] Example 16: HC-5404 enhances antiangiogenic agents in a 786-O RCC xenograft model

[0572] In this study, we investigated the inhibition of PERK-mediated stress adaptation using HC-5404, which was combined with a VEGRF-TKI to inhibit tumor growth in the 786-O xenograft RCC model.

[0447]

[0573] Following similar procedures as described in the above examples, 786-O tumor xenografts were treated with oral lenvatinib (10 mg / kg) once daily, oral HC-5404 (30 mg / kg) twice daily, or a combination thereof, and oral axitinib (30 mg / kg) twice daily, oral HC-5404 (30 mg / kg) twice daily, or a combination thereof. Tumor growth curves are shown in Figures 48 and 49. Following similar procedures as described in the above examples, A498 tumor xenografts were treated with oral sunitinib (20 mg / kg) once daily, oral HC-5404 (30 mg / kg) twice daily, or a combination thereof, and twice-weekly intraperitoneal injections of DC-101 (15 mg / kg), oral HC-5404 (30 mg / kg) twice daily, or a combination thereof. Tumor growth curves are shown in Figures 50 and 51.

[0448]

[0574] Following similar procedures as described in the previous examples, 786-O tumor xenografts were treated with twice-daily oral doses of 3, 10, or 30 mg / kg HC-5404, either as a single agent or in combination with a once-daily oral dose of 30 mg / kg cabozantinib. Tumor growth and body weight curves are shown in Figures 52 and 53. HC-5404 at 3, 10, and 30 mg / kg PO or BID demonstrated a dose-dependent trend toward antitumor efficacy, with maximal efficacy at 30 mg / kg (TGI% of approximately 61%), while cabozantinib monotherapy resulted in tumor stagnation and only 2 / 8 PRs in this model. Combination treatment with 10 and 30 mg / kg HC-5404 significantly improved the efficacy of cabozantinib alone, demonstrating similar efficacy with 8 / 8 PRs in each group and a mean tumor regression of approximately 66%. No treatment-related weight loss was observed. Treatment with HC-5404 or cabozantinib, either as monotherapy or in combination, did not affect the body weight of the mice, suggesting that the treatments were well tolerated.

[0449] Example 17: Cabozantinib inhibits the formation of new tumor vasculature and destroys pericytes

[0575] In this example, the inhibitory effect of cabozantinib on the formation of new tumor vasculature and pericytes was investigated following a similar procedure as described above.

[0450]

[0576] Cabozantinib induced a near-complete loss of immature vasculature (CD31+SMA- cells). HC-5404 reduced mature CD31+SMA+ vascular cells, a trend seen only in the presence of cabozantinib. Neovascularization is highly dependent on VEGF signaling. IHC staining images are shown in Figure 54, and quantification of IHC staining is shown in Figure 55.

[0451]

[0577] Pericytes are a stabilizing cell layer and are associated only with mature blood vessels. The combination of HC-5404 and cabozantinib reduced pericyte markers NG2 and MCAM. Statistical comparisons only showed significant differences between the vehicle and treatment groups. A trend toward MCAM was observed for the cabozantinib + HC-5404 group. IHC staining images are shown in Figure 56, and quantification of IHC staining is shown in Figure 57.

[0452] Example 18: Triple drug combination induces anti-tumor immune responses

[0578] In this example, we investigated the efficacy of tumor treatment with a combination of HC-5404, an antiangiogenic agent, and immunotherapy, following a similar procedure as described above. In RENCA tumor-bearing BALB / c mice, HC-5404 showed improved efficacy in combination with axitinib and an anti-PD-1 antibody.

[0453]

[0579] Mice were inoculated subcutaneously with syngeneic RENCA RCC tumor cells. Six days after cell inoculation, mice were randomized to achieve equivalent starting tumor size and then initiated the indicated treatment. Tumor volume (Figure 58) and body weight (Figure 59) were monitored over time and are shown. PO = oral gavage; IV = intravenous; BID = twice daily; BIW = twice week...

Claims

1. 1. A method of activating the immune system of a subject in need thereof, comprising administering to said subject a therapeutically effective amount of a compound of formula (I) 【Chemical 1】 or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of an anti-PD-1 antibody.

2. 1. A method of activating the immune system of a subject having cancer, comprising administering to said subject a therapeutically effective amount of a compound of formula (I) 【Chemistry 2】 or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of an anti-PD-1 antibody.

3. The method of claim 1 or 2, wherein the activity of myeloid-derived suppressor cells (MDSCs) is inhibited.

4. The method according to any one of claims 1 to 3, which increases the infiltration of T cells and NK cells into tumors.

5. 5. The method of claim 4, wherein the tumor is selected from the group consisting of bladder cancer, head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma.

6. The method of any one of claims 1 to 5, wherein the frequency of dendritic cells in draining lymph nodes is increased.

7. A method of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formula (I) 【Chemistry 3】 or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of an anti-PD-1 antibody.

8. The method of any one of claims 2 to 7, wherein the cancer is a solid tumor.

9. The method of any one of claims 2 to 4 or 6 to 8, wherein the cancer is selected from the group consisting of bladder cancer, head and neck squamous cell carcinoma, Merkel cell carcinoma, hepatocellular carcinoma, renal cell carcinoma, cervical cancer, small cell lung cancer, non-small cell lung cancer, triple-negative breast cancer, gastric cancer, endometrial cancer, urothelial carcinoma, and neuroendocrine carcinoma.

10. The method of any one of claims 2 to 4, 6 or 7, wherein the cancer is a liquid tumor.

11. 11. The method of any one of claims 2-4, 6, 7 or 10, wherein the cancer is selected from the group consisting of classical Hodgkin's lymphoma, primary thymic-mediastinal lymphoma, multiple myeloma, and B-cell malignancies selected from non-Hodgkin's lymphoma and chronic lymphocytic leukemia.

12. The method of any one of claims 1 to 11, wherein the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, and cemiplimab.

13. The method of any one of claims 1 to 12, wherein the anti-PD-1 antibody is pembrolizumab.

14. The method of any one of claims 1 to 12, wherein the anti-PD-1 antibody is nivolumab.

15. 15. The method of any one of claims 1 to 14, wherein the therapeutically effective amount of the anti-PD-1 antibody is administered once daily.

16. 15. The method of any one of claims 1 to 14, wherein the therapeutically effective amount of the anti-PD-1 antibody is administered twice daily.

17. 17. The method of any one of claims 1 to 16, wherein the therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof and the therapeutically effective amount of the anti-PD-1 antibody are administered simultaneously.

18. 17. The method of any one of claims 1 to 16, wherein the therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof and the therapeutically effective amount of the anti-PD-1 antibody are administered sequentially.

19. 19. The method of any one of claims 1 to 18, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered twice daily in an amount of about 22 mg to about 451 mg.

20. 19. The method of any one of claims 1 to 18, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered twice daily in an amount of about 177 mg to about 900 mg.

21. The method of any one of claims 1 to 20, wherein the therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered orally.

22. The method of any one of claims 1 to 21, wherein the therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in the form of a capsule.

23. 23. The method of any one of claims 1 to 22, wherein the method increases tumor cell expression of type 1 interferon receptor (INFAR1) in the subject relative to a subject receiving a placebo, the therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof, or the therapeutically effective amount of the anti-PD-1 antibody.

24. 24. The method of any one of claims 1 to 23, wherein the method increases peripheral blood monocyte surface expression of INFAR1 in the subject relative to a subject receiving a placebo, the therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof, or the therapeutically effective amount of the anti-PD-1 antibody.

25. 25. The method of any one of claims 1 to 24, wherein the method increases tumor cell expression of calreticulin in the subject relative to a subject receiving a placebo, the therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof, or the therapeutically effective amount of the anti-PD-1 antibody.

26. 26. The method of any one of claims 1 to 25, wherein the method increases polymorphonuclear myeloid-derived suppressor cell (MDSC) and / or tumor-associated macrophage (TAM) expression of INFAR1 in the subject compared to a subject receiving a placebo, the therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof, or the therapeutically effective amount of the anti-PD-1 antibody.

27. 27. The method of any one of claims 1 to 26, wherein the method increases TAM expression of PD-L1 in the subject relative to a subject receiving a placebo, the therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof, or the therapeutically effective amount of the anti-PD-1 antibody.

28. 28. The method of any one of claims 1 to 27, wherein the method increases infiltration of CD8 T cells and / or NK cells into tumors in the subject compared to a subject receiving a placebo, the therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof, or the therapeutically effective amount of the anti-PD-1 antibody.

29. 29. The method of any one of claims 1-28, wherein the method increases expression of CD69 on T cells in tumor-draining lymph nodes in a subject relative to a subject receiving a placebo, the therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof, or the therapeutically effective amount of the anti-PD-1 antibody.

30. The method of any one of claims 1 to 29, wherein the suppressive activity of MDSCs in the subject is reduced.

31. The method of any one of claims 1 to 30, wherein the method increases the frequency of dendritic cells in tumor-draining lymph nodes in the subject.

32. 32. The method of any one of claims 1 to 31, wherein Ki67, GzmB and memory phenotype markers are increased on CD4 T cells in the subject.

33. 33. The method of any one of claims 1 to 32, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered as a pharmaceutically acceptable salt of the compound of formula (I).

34. 34. The method of claim 33, wherein the pharmaceutically acceptable salt is a hemifumarate salt.

35. 1. A method of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formula (I): 【Chemistry 4】 or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of an anti-angiogenic agent.

36. 1. A method of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formula (I): 【Chemistry 5】 or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an anti-angiogenic agent, wherein the subject is being treated with an anti-cancer therapy.

37. 37. The method of claim 36, wherein the subject is resistant or has acquired resistance to the anti-cancer therapy.

38. 37. The method of claim 36, wherein the anti-cancer therapy is selected from the group consisting of administration of an immunotherapeutic agent, a chemotherapeutic agent, a growth inhibitory agent, a cytotoxic agent, an anti-angiogenic agent, or a combination thereof, radiation therapy, surgery, and a combination thereof.

39. 39. The method of any one of claims 35 to 38, wherein the therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof and the therapeutically effective amount of the anti-angiogenic agent are administered simultaneously.

40. 39. The method of any one of claims 35 to 38, wherein the therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof and the therapeutically effective amount of the anti-angiogenic agent are administered sequentially.

41. 41. The method of any one of claims 35 to 40, wherein the therapeutically effective amount of the anti-angiogenic agent is administered to the subject daily.

42. 42. The method of any one of claims 35 to 41, wherein the therapeutically effective amount of the anti-angiogenic agent is administered to the subject once daily.

43. 42. The method of any one of claims 35 to 41, wherein the therapeutically effective amount of the anti-angiogenic agent is administered to the subject twice daily.

44. 44. The method of any one of claims 35 to 43, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of the anti-angiogenic agent reduces p-PERK levels in the subject compared to administering a therapeutically effective amount of the anti-angiogenic agent alone.

45. 44. The method of any one of claims 35 to 43, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of the anti-angiogenic agent increases ATF4 levels in the subject compared to administering a therapeutically effective amount of the anti-angiogenic agent alone.

46. 44. The method of any one of claims 35 to 43, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of the anti-angiogenic agent modulates activation of the unfolded protein response (UPR) compared to administering a therapeutically effective amount of the anti-angiogenic agent alone.

47. 1. A method of treating cancer in a subject receiving a therapeutically effective amount of anti-angiogenic therapy, comprising administering to the subject a therapeutically effective amount of a compound of formula (I): 【Chemistry 6】 or a pharmaceutically acceptable salt thereof.

48. 48. The method of claim 47, wherein the subject is resistant or has acquired resistance to the antiangiogenic agent therapy.

49. 49. The method of any one of claims 35 to 48, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof reduces p-PERK levels in the subject compared to administering a therapeutically effective amount of the anti-angiogenic agent alone.

50. 49. The method of any one of claims 35 to 48, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof increases ATF4 levels in the subject compared to administering a therapeutically effective amount of the anti-angiogenic agent alone.

51. 49. The method of any one of claims 35 to 48, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof modulates activation of the unfolded protein response (UPR) compared to administering a therapeutically effective amount of the anti-angiogenic agent alone.

52. 49. The method of any one of claims 35 to 48, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof reduces tumor vasculature in the subject compared to administering a therapeutically effective amount of the anti-angiogenic agent alone.

53. 53. The method of any one of claims 35 to 52, wherein the anti-angiogenic agent is a vascular endothelial growth factor receptor (VEGFR) modulator.

54. 53. The method of any one of claims 35 to 52, wherein the anti-angiogenic agent is a vascular endothelial growth factor (VEGF)-targeting antibody.

55. The method of any one of claims 35 to 52, wherein the anti-angiogenic agent is a tyrosine kinase inhibitor (TKI).

56. The method according to any one of claims 35 to 52, wherein the anti-angiogenic agent is a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI).

57. 57. The method of any one of claims 35 to 56, wherein the antiangiogenic agent is selected from the group consisting of apatinib, axitinib, brivanib, cabozantinib, canertinib, cediranib, dasatinib, erlotinib, gefitinib, leflunomide, lenvatinib, motesanib, nilotinib, nintedanib, pazopanib, regorafenib, semaxinib, sorafenib, sunitinib, tivozanib, vandetanib, vatalanib, XL0192, bevacizumab, ramucirumab, and pharmaceutically acceptable salts or biosimilars thereof.

58. 57. The method of any one of claims 35 to 56, wherein the antiangiogenic agent is selected from the group consisting of bevacizumab, ramucirumab, and biosimilars thereof.

59. 57. The method of any one of claims 35 to 56, wherein the antiangiogenic agent is selected from the group consisting of apatinib, axitinib, brivanib, cabozantinib, canertinib, cediranib, dasatinib, erlotinib, gefitinib, leflunomide, lenvatinib, motesanib, nilotinib, nintedanib, pazopanib, regorafenib, semaxinib, sorafenib, sunitinib, tivozanib, vandetanib, vatalanib, XL0192, and pharmaceutically acceptable salts thereof.

60. 60. The method of any one of claims 35 to 59, wherein the cancer is metastatic cancer or locally advanced cancer.

61. 61. The method of any one of claims 35 to 60, wherein the cancer is a solid tumor.

62. 62. The method of any one of claims 35 to 61, wherein the cancer is selected from the group consisting of bladder cancer, breast cancer, carcinoma, cervical cancer, colorectal cancer, gastric cancer, hepatocellular carcinoma, renal cancer, lung cancer, neuroendocrine cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer and thyroid cancer.

63. 63. The method of claim 62, wherein the breast cancer is triple-negative breast cancer or metastatic breast cancer, the carcinoma is carcinoma of unknown primary (CUP), endometrial cancer, head and neck squamous cell carcinoma, Merkel cell carcinoma, squamous cell carcinoma or urothelial carcinoma, the gastric cancer is adenocarcinoma or gastrointestinal stromal tumor, the kidney cancer is renal cell carcinoma (RCC), or the lung cancer is small cell lung cancer (SCLC) or non-small cell lung cancer.

64. 63. The method of claim 62, wherein the renal cell carcinoma is clear cell renal cell carcinoma (ccRCC).

65. 63. The method of claim 62, wherein the cancer is selected from the group consisting of breast cancer, gastric cancer, renal cancer, and lung cancer.

66. 66. The method of any one of claims 35 to 65, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject from about 22 mg to about 798 mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof.

67. 67. The method of any one of claims 35 to 66, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject from about 22 mg to about 798 mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof.

68. 67. The method of any one of claims 35 to 66, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject from about 22 mg to about 798 mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof daily.

69. 67. The method of any one of claims 35 to 66, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject from about 22 mg to about 798 mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof once daily.

70. 67. The method of any one of claims 35 to 66, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject from about 22 mg to about 798 mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof twice daily.

71. 67. The method of any one of claims 35 to 66, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof comprises administering to the subject from about 44 mg to about 177 mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof.

72. 67. The method of any one of claims 35 to 66, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject from about 44 mg to about 177 mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof.

73. 67. The method of any one of claims 35 to 66, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject from about 44 mg to about 177 mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof daily.

74. 67. The method of any one of claims 35 to 66, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject from about 44 mg to about 177 mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof once daily.

75. 67. The method of any one of claims 35 to 66, wherein administering a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof comprises orally administering to the subject from about 44 mg to about 177 mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof twice daily.

76. 76. The method of any one of claims 35 to 75, wherein the subject is in a fasting state.

77. 76. The method of any one of claims 35 to 75, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered with food.

78. 76. The method of any one of claims 35 to 75, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered about 30 minutes to about 1 hour after a meal.

79. The method of any one of claims 35 to 78, wherein the compound of formula (I) is administered as a pharmaceutically acceptable salt.

80. 80. The method of claim 79, wherein the pharmaceutically acceptable salt is a hemifumarate salt.

81. 81. The method of any one of claims 35 to 80, further comprising administering to the subject a therapeutically effective amount of immunotherapy.

82. 1. A method of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formula (I): 【Chemistry 7】 or a pharmaceutically acceptable salt thereof, a therapeutically effective amount of an anti-angiogenic agent, and a therapeutically effective amount of an immunotherapy.

83. 1. A method of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formula (I): 【Chemistry 8】 or a pharmaceutically acceptable salt thereof, a therapeutically effective amount of an anti-angiogenic agent, and a therapeutically effective amount of an immunotherapy, wherein the subject is being treated with an anti-cancer therapy.

84. 84. The method of claim 83, wherein the subject is resistant or has acquired resistance to the anti-cancer therapy.

85. 84. The method of claim 83, wherein the anti-cancer therapy is selected from the group consisting of surgery, radiation therapy, administration of an immunotherapeutic agent, a chemotherapeutic agent, a growth inhibitory agent, a cytotoxic agent, an anti-angiogenic agent, or a combination thereof, and combinations thereof.

86. 1. A method of treating cancer in a subject receiving a therapeutically effective amount of anti-angiogenic therapy, comprising administering to the subject a therapeutically effective amount of a compound of formula (I): 【Chemistry 9】 or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of immunotherapy.

87. 87. The method of any one of claims 81 to 86, wherein the immunotherapy is an immune checkpoint inhibitor.

88. The method of any one of claims 81 to 86, wherein the immunotherapy is an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody.

89. 89. The method of claim 88, wherein the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, and cemiplimab.

90. 90. The method of claim 88 or 89, wherein the anti-PD-1 antibody is pembrolizumab.

91. 90. The method of claim 88 or 89, wherein the anti-PD-1 antibody is nivolumab.

92. 92. The method of any one of claims 81-91, wherein the therapeutically effective amount of immunotherapy is administered daily.

93. 92. The method of any one of claims 81-91, wherein the therapeutically effective amount of immunotherapy is administered once daily.

94. 92. The method of any one of claims 81-91, wherein the therapeutically effective amount of immunotherapy is administered twice daily.

95. 95. The method of any one of claims 81 to 94, wherein the therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof and the therapeutically effective amount of immunotherapy are administered simultaneously.

96. 95. The method of any one of claims 81 to 94, wherein the therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof and the therapeutically effective amount of immunotherapy are administered sequentially.

97. 95. The method of any one of claims 81 to 94, wherein the therapeutically effective amount of the anti-angiogenic agent and the therapeutically effective amount of immunotherapy are administered simultaneously.

98. 95. The method of any one of claims 81 to 94, wherein the therapeutically effective amount of the anti-angiogenic agent and the effective amount of immunotherapy are administered sequentially.

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