Combination for cancer treatment

Abstract

The present invention relates to a combination comprising a checkpoint inhibitor and a c-Met inhibitor, and compound 1. The present invention also relates to a crystalline form of the free base of compound 1 combined with a checkpoint inhibitor, as well as a crystalline form of the salt of compound 1. The present invention also relates to a pharmaceutical composition comprising such combinations. The present invention further relates to a method for treating cancer by administering compound 1 as a single agent or as a combination described herein.

Description

Technology Field

[0001] Cross-reference regarding related applications

[0002] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 148,921 filed February 12, 2021, U.S. Provisional Application No. 63 / 113,556 filed November 13, 2020, and U.S. Provisional Application No. 63 / 059,601 filed July 31, 2020, all of which are incorporated herein by reference in their entirety.

[0003] Technical field of the present invention

[0004] The present invention relates to a combination comprising an immune checkpoint inhibitor (ICI) and Compound 1. The present invention also relates to a crystalline form of the free base of Compound 1 combined with a checkpoint inhibitor, as well as a crystalline form of the salt of Compound 1. The present invention also relates to a pharmaceutical composition of Compound 1 used in combination with a checkpoint inhibitor. The present invention further relates to a method for treating cancer by administering Compound 1 as a single agent or as a combination as described herein. Background Technology

[0005] Cancer is a significant cause of morbidity and mortality worldwide. While standard treatments for many different types of cancer have improved significantly over the years, current standard therapies still fail to meet the demand for effective therapies to improve cancer treatment. Standard treatments for many cancer types have been improved by the clinical use of immuno-oncology agents targeting cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), programmed cell death receptor-1 (PD-1), and its ligand PD-L1. Although these checkpoint inhibitors demonstrate improved clinical responses in certain of these cancers, sustained clinical responses occur in only approximately 10% to 45% of patients. Furthermore, a significant number of tumors become resistant or refractory.

[0006] In recent years, TAM tyrosine kinases, particularly AXL receptor tyrosine kinases, have emerged as promising targets for cancer treatments. AXL is a cell surface receptor tyrosine kinase that is part of the TAM kinase family, which includes TYRO3 and MERTK. Several drugs classified as "AXL inhibitors" have entered clinical trials; however, the majority target multiple kinase receptors in addition to AXL.

[0007] Human Axl belongs to the Tyro3, ​​Axl, and Mer (TAM) subfamily of receptor tyrosine kinases that includes Mer. TAM kinases are characterized by an extracellular ligand binding domain consisting of two immunoglobulin-like domains and two fibronectin type III domains. Axl is overexpressed in numerous tumor cell types and was initially cloned from patients with chronic myeloid leukemia. When overexpressed, Axl exhibits transforming ability. Axl signaling is believed to induce tumor growth through the activation of proliferation and anti-apoptotic signaling pathways. Axl is associated with cancers such as lung cancer, myeloid leukemia, uterine cancer, ovarian cancer, glioma, melanoma, thyroid cancer, renal cell carcinoma, osteosarcoma, gastric cancer, prostate cancer, and breast cancer. Overexpression of Axl is associated with a poor prognosis in patients with the specified cancers.

[0008] Similar to Axl, the activation of Mer transmits downstream signaling pathways that induce tumor growth and activation. Mer binds to ligands such as the soluble protein Gas-6. Gas-6 binding to Mer induces autophosphorylation in Mer's intracellular domain, leading to downstream signal activation. In cancer cells, the overexpression of Mer likely increases metastasis through the production of soluble Mer extracellular domain proteins acting as attractant receptors. Tumor cells secrete soluble forms of extracellular Mer receptors that reduce the ability of soluble Gas-6 ligands to activate Mer in endothelial cells, leading to cancer progression.

[0009] Accordingly, there is a demand in the industry for new therapies, including combination therapies for the treatment of cancer, for example. Solutions to these and other problems in the industry are provided in this specification.

[0010] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising the following steps:

[0011] (i) a step of administering to a subject a dose of about 5 mg to about 100 mg of compound 1 or a pharmaceutically acceptable salt thereof, wherein compound 1 has the following structure: said administering step:

[0012] , and

[0013] (ii) A step of administering a therapeutically effective amount of a checkpoint inhibitor to the subject.

[0014] In another aspect, the present invention comprises a method for treating cancer in a subject, wherein the method comprises the following compound 1 in a dose of about 5 mg to about 100 mg:

[0015]

[0016] or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient,

[0017] The method includes the step of administering a therapeutically effective amount of a gate inhibitor or a pharmaceutical composition containing a gate inhibitor to a subject requiring such treatment.

[0018] In one embodiment, the present invention comprises a method for treating urothelial carcinoma in a subject, the method comprising the following steps:

[0019] (i) a step of administering to a subject a dose of about 5 mg to about 100 mg of compound 1 or a pharmaceutically acceptable salt thereof, wherein compound 1 has the following structure: said administering step:

[0020] , and

[0021] (ii) A step of administering a therapeutically effective amount of a checkpoint inhibitor to the subject.

[0022] In another aspect, the present invention comprises a method for treating urothelial carcinoma in a subject, wherein the method comprises the following compound 1 in a dose of about 5 mg to about 100 mg:

[0023]

[0024] or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient,

[0025] The method includes the step of administering a therapeutically effective amount of a gate inhibitor or a pharmaceutical composition containing a gate inhibitor to a subject requiring such treatment.

[0026] In these and other embodiments, the checkpoint inhibitor is selected from the group consisting of PD-1 inhibitors, PD-L1 inhibitors, and CTLA-4 inhibitors. In these and other aspects and embodiments, the checkpoint inhibitor is selected from the group consisting of pembrolizumab, nivolumab, atezolizumab (TECENTRIQ®), durvalumab, avelumab (BAVENCIO®), cemiplimab, camrelizumab, syntilimab, tisraelizumab, toripalizumab, spartalizumab, dostalimab, KN035 (Jiangsu Alphamb Biopharmaceuticals Co.), cocibelimab (formerly CK-301), CA-170 (Curis, Inc.), BMS-986189 (Bristol Myers Squibb Co.), and ipilimumab (Yervoy, Bristol Myers Squibb Co.).

[0027] In a further embodiment, a method for treating cancer in a subject comprising the following is provided:

[0028] (i) a step of administering to a subject a dose of about 5 mg to about 100 mg of compound 1 or a pharmaceutically acceptable salt thereof, wherein compound 1 has the following structure: said administering step:

[0029] , and

[0030] (ii) a step of administering a therapeutically effective amount of nivolumab and at least one additional immunomodulator to the subject.

[0031] In one embodiment of this aspect, the immunomodulator is selected from the group consisting of PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, and IL-2 targeting agents.

[0032] In these and other modes, the subject is a human subject in need of treatment.

[0033] In these and other embodiments, the checkpoint inhibitor is selected from the group consisting of PD-1 inhibitors, PD-L1 inhibitors, and CTLA-4 inhibitors. In these and other aspects and embodiments, the checkpoint inhibitor is selected from the group consisting of pembrolizumab, nivolumab, atezolizumab (TECENTRIQ®), durvalumab, avelumab (BAVENCIO®), cemiplimab, camrelizumab, syntilimab, tisraelizumab, toripalizumab, spartalizumab, dostalimab, KN035 (Jiangshu Alphamab Biopharmaceuticals Company), cocibelimab (formerly CK-301), CA-170 (Kuris, Inc.), BMS-986189 (Bristol Myers Squibb Company), and ipilimumab (Yervoy, Bristol Myers Squibb Company).

[0034] In these and other embodiments, IL-2 targeting agents are CD122-preferential IL-2 pathway agonists, PEG-IL-2Rαβ-biased agonists, IL-2Rβ-biased agonists, IL-2Rβγ c- Biased agonist, IL-2v / IL-2α fusion protein, anti-EDB mAb(L19) / IL-2v fused to L19 / TNFv, anti-GD2 mAb / IL-2v, anti-FAP mAb / IL-2v, anti-CEA mAb / IL-2v, anti-PD-1 mAb / IL-2v, vaccine of patient-derived tumor cells + HD-IL-2, adoptive cell therapy + IL-2 infusion, adoptive cell therapy + IL-2 infusion + anti-PD-1 mAb, orthogonal IL-2v / IL-2Rβ mutant pair, anti-IL-2Rα mAb / PBD conjugate, PEG-IL-2Rα-biased agonist, IL-2v / human Fc fusion protein, PEG-IL-2Rα-biased(N88D) / IgG1 fusion protein, anti-IL-2 mAb / IL-2v, IL-2, It is selected from the group consisting of PPIs, recombinant plasmids encoding TGF-β1 and IL-10, and IL-2Rβ antagonists.

[0035] In one embodiment, the IL-2 targeting agent is a CD122-preferential IL-2 pathway agonist. In one embodiment, the CD122-preferential IL-2 pathway agonist is bempegaldesleukin (BEMPEG; NKTR-214; Bristol Myers Squibb Company).

[0036] In one embodiment, the IL-2 targeting agent is a PEG-IL-2Rα-biased agonist. In one embodiment, the PEG-IL-2Rα-biased agonist is NKTR-358 (Bristol Myers Squibb Company).

[0037] In a further embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising the step of administering to the subject a dose of about 5 mg to about 100 mg of compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1.

[0038] In these and other embodiments, Compound 1 is administered as a free base crystalline solid or a crystalline pharmaceutically acceptable salt. To avoid any doubt, “Compound 1” refers to the crystalline salt form as well as this crystalline free base form unless otherwise indicated.

[0039] In these and other embodiments, compound 1 is a crystalline solid form characterized by form A, form B, form C, form D, form E, form F, form G, form H, form K, form O or form Q.

[0040] In these and other embodiments, compound 1 is a crystalline HCl salt of compound 1.

[0041] In these and other embodiments, compound 1 is a crystalline fumarate of compound 1, or a hydrate or solvate thereof.

[0042] In these and other embodiments, compound 1 is a crystalline phosphate of compound 1, or a hydrate or solvate thereof. Brief explanation of the drawing

[0043] Figure 1A shows the presence of tumor microvessels by CD31 staining after treatment with compound 1. Horizontal bars represent the average values ​​for n=3 tumors per condition. Figure 1B shows the presence of tumor microvessels by CD31 staining after treatment with compound 1. Paraffin-embedded tumor tissue was stained with the vascular marker CD31 and scored for vascular density according to condition. Figure 2A shows the presence of cytotoxic T-cells by CD8 staining after treatment with the combination of compound 1, PD-1, and compound 1 + PD-1. Horizontal bars represent the median for n=8-12 tumors per condition. Figure 2B shows the presence of cytotoxic T-cells by CD8 staining after treatment with PD-1 and a combination of compound 1 + PD-1. Paraffin-embedded tumor tissues were stained with the vascular marker CD8 and scored for vascular density according to condition. Figures 3A to 3C show tumor volume after treatment with combination therapy of compound 1 + PD-1, compound 1 + PD-L1, and compound 1 + CTLA-4. Figure 4 shows the growth curve of CT26 colon cells transplanted subcutaneously in mice treated with compound 1 as a single agent or in combination with an anti-PD-1 inhibitor (administration period 40 days). Figure 5 shows the Kaplan-Meier survival curves of CT26 colon tumor-bearing mice treated with a combination of compound 1, an anti-PD-1 inhibitor, and compound 1 + an anti-PD-1 inhibitor (administration period 40 days). Figure 6A compares tumor growth after treatment with vehicle, 30 mg / kg compound 1, 10 mg / kg anti-PD-1, or both. Symbols indicate median tumor volume. Figure 6B shows a Kaplan-Meier plot showing conditional survival for CT26 tumor-bearing mice after treatment. For conditional survival, 40% of the animals in the treatment group were excluded from the study when they reached the tumor size threshold. Figure 7A shows that compound 1 inhibits efferocytosis in a dose-dependent manner using 25k apoptotic Jurkat. Figure 7B shows that compound 1 inhibits the removal of apoptotic cells in a dose-dependent manner using 50k apoptotic Jurkat. Figure 8 shows the mean (SD) plasma concentration-time profile of Compound 1 as a single agent after the first administration and after the 28th day administration in subjects with solid tumors. Specific details for implementing the invention

[0044]

[0045]

[0046]

[0047]

[0048] As used herein, the following definitions apply unless otherwise indicated.

[0049] For the purposes of the present invention, chemical elements are identified according to the periodic table of elements in the CAS version of the Handbook of Chemistry and Physics (95th Ed). Additionally, general principles of organic chemistry are referenced in the literature ["Organic Chemistry," 2, the entire contents of which are incorporated herein by reference. nd It is described in [Ed., Thomas Sorrell, University Science Books, Sausalito: 2006, and "March's Advanced Organic Chemistry," 7th Ed., Ed.: Smith, MB and March, J., John Wiley & Sons, New York: 2013].

[0050] As used herein, the term “low / limited / significant hygroscopicity” refers to a material that exhibits moisture absorption of < 0.5 / < 2.0 / ≥ 2.0 wt% in a specified RH range.

[0051] As used herein, the term "stoichiometric hydrate" refers to a crystalline material having a water content defined over an extended RH range. Typical stoichiometric hydrates include hemihydrates, monohydrates, sesquihydrates, dihydrates, etc.

[0052] The term “variable hydrate” as used in this specification refers to a crystalline material having varying moisture content over an extended RH range but without phase change.

[0053] As used herein, the chemical term designated as "form" refers to a compound consisting of a single phase or a salt thereof.

[0054] The term “low / limited / intermediate / good / high solubility” as used in this specification refers to a substance having a solubility of < 1 mg / ml / 1 mg / ml to 20 mg / ml / 20 mg / ml to 100 mg / ml / 100 mg / ml to 200 mg / ml / > 200 mg / ml.

[0055] As used herein, the term "crystalline" refers to a material that produces an XRPD pattern having sharp peaks (similar to the peak width of the instrument) and weak diffuse scattering relative to the peaks.

[0056] As used herein, the term "disordered crystalline" refers to a material that produces an XRPD pattern having broad peaks (relative to the instrument peak width) and / or strong diffuse scattering relative to the peaks. A disordered material is:

[0057] 1) Microcrystalline,

[0058] 2) Crystalline with high defect density,

[0059] 3) A mixture of crystalline and X-ray amorphous phases or

[0060] 4) The above combination

[0061] It could be.

[0062] As used herein, the term "insufficient signal" means that the spectroscopic analysis of a sample has generated a spectrum or pattern (output) having an insufficient signal greater than the expected background noise.

[0063] As used herein, the term "single crystalline phase" refers to an XRPD pattern determined to contain evidence of a single crystalline form due to Bragg peaks indexed to a single unit cell. Indexing is the process of assigning a Miller index label to each peak of a diffraction pattern. Additionally, the size and shape of the crystalline unit cell are determined during the indexing process.

[0064] As used herein, the term “slurry” refers to a suspension prepared by adding a sufficient amount of solid to a given solvent under ambient conditions such that undissolved solid is present. A typical slurry involves stirring (typically by stirring or vibration), which is an act also referred to as “slurrying,” in a sealed vial at a given temperature for an extended period. Typically, the solid is recovered after a given period using the method described herein.

[0065] As used herein, the terms "X-ray amorphous" or "amorphous" refer to a material in which diffuse scattering is present but there is no evidence of Bragg peaks in the XRPD pattern.

[0066] As used herein, the term “crystalline” refers to a solid-state compound having a periodic and repeating three-dimensional internal arrangement of atoms, ions, or molecules that is characteristic of a crystal arranged in a lattice having a fixed geometric pattern or strict long-range order, for example. The term crystalline means that the compound does not necessarily exist as a crystal, but has such a crystal-like internal structural arrangement.

[0067] As used herein, the term “substantially crystalline” refers to a solid material that is primarily arranged in a fixed geometric pattern or lattice having strict long-range order. For example, a substantially crystalline material has a crystallinity greater than about 85% (e.g., crystallinity greater than about 90%, crystallinity greater than about 95%, or crystallinity greater than about 99%). It should be noted that the term “substantially crystalline” includes the predicate “crystalline” as defined in the preceding paragraph.

[0068] For the purposes of the present invention, "patient" includes humans and any other animals, particularly mammals and other organisms. Accordingly, the method is applicable to both human treatment and veterinary use. In a preferred embodiment, the patient is a mammal, and in the most preferred embodiment, the patient is a human. Examples of preferred mammals include mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, and primates.

[0069] "Kinese-dependent disease or condition" refers to a pathological condition that depends on the activity of one or more kinases. Kinases participate directly or indirectly in the signaling pathways of various cellular activities, including proliferation, adhesion, migration, differentiation, and invasion. Diseases related to kinase activity include tumor growth, pathological neovascularization supporting solid tumor growth, and other diseases involving excessive local angiogenesis such as ocular diseases (diabetic retinopathy, age-related macular degeneration, etc.) and inflammation (psoriasis, rheumatoid arthritis, etc.).

[0070] "Therapeutically effective amount" is an amount of the crystalline form or crystalline salt form of the present invention that improves the symptoms of the disease. The amount of the crystalline form or crystalline salt form of the present invention constituting the "therapeutic effective amount" may vary depending on the compound, the state of the disease and its severity, the age of the patient to be treated, etc. The therapeutically effective amount may be routinely determined by a person skilled in the art in consideration of the knowledge of a person skilled in the art and the present disclosure.

[0071] The phrase “pharmaceuticalally acceptable” is used herein to refer to compounds, substances, compositions, and / or dosage forms corresponding to a reasonable benefit-risk ratio within the scope of reasonable medical judgment that are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, immunogenicity, or other problems or complications.

[0072] As used herein, the term “pharmaceuticalally acceptable excipient” refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, solvent, or encapsulating material. Excipients include those that are generally safe, non-toxic, biologically undesirable, and acceptable for human pharmaceutical uses as well as veterinary uses. In one embodiment, each component is “pharmaceutically acceptable” as defined herein. For example, the literature [Remington: The Science and Practice of Pharmacy, 21st ed.; Lippincott Williams & Wilkins: Philadelphia, Pa., 2005; Handbook of 'Pharmaceutical Excipients, 6th ed.; Rowe et al, Eds.; The Pharmaceutical Press and the American Pharmaceutical Association: 2009; Handbook of Pharmaceutical Additives, 3rd ed. ; Ash and Ash Eds. ; Gower Publishing Company: 2007; Pharmaceutical Pref or mulation and Formulation, 2nd ed.; Gibson Ed.; CRC Press LLC: Boca Raton, Fla., 2009.

[0073] As used herein, the term "concurrently" means simultaneously. For example, if two therapeutic regimens are performed concurrently for a single patient, they are being performed simultaneously. It will be understood that the simultaneous occurrence of two therapeutic regimens does not necessarily imply that the actual delivery of two drugs occurs simultaneously, as each regimen may require a different administration schedule and / or a different delivery method.

[0074] As used herein and provided by the National Cancer Institute, “checkpoint inhibitor” refers to any agent that blocks, inhibits, or modulates a checkpoint protein. Checkpoint inhibitors are produced by some types of immune system cells, such as T cells and some cancer cells. Examples of checkpoint proteins found in T cells or cancer cells include PD-1 / PD-L1 and CTLA-4 / B7-1 / B7-2. Examples of checkpoint inhibitors include pembrolizumab, nivolumab, atezolizumab, durvalumab, avelumab, cemiflimab, camrelizumab, syntilimab, tisraelizumab, toripalizumab, spartalizumab, dostalimab, KN035 (Jiangshu Alphamab Biopharmaceuticals Company), cosivelimab (formerly CK-301), CA-170 (Kuris, Inc.), and BMS-986189 (Bristol Myers Squibb Company). Examples of FDA-approved checkpoint inhibitors include pembrolizumab, nivolumab, atezolizumab, durvalumab, avelumab, and cemiflimab products. Dosage and other information regarding approved checkpoint inhibitors are available from the FDA, EMEA, or other national healthcare regulatory agencies.

[0075] "Cancer" refers to any physiological condition in mammals characterized by uncontrolled cell growth; in particular, a cell-proliferative disorder state including, but not limited to, the following: heart : Sarcomas (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyosarcoma, fibroma, lipoma, and teratoma; Head and neck: Squamous cell carcinoma of the head and neck, laryngeal and hypopharyngeal cancer, nasal and paranasal sinus cancer, nasopharyngeal cancer, salivary gland cancer, oral and pharyngeal cancer; lung : Bronchial carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma, non-small cell lung cancer), alveolar (bronchiolar) carcinoma, alveolar sarcoma, alveolar soft tissue sarcoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma; colon:Colorectal cancer, adenocarcinoma, gastrointestinal stromal tumor, lymphoma, carcinoid, Turcot syndrome; camouflage : Gastric cancer, gastroesophageal junction adenocarcinoma, esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, non-forma), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma); breast: Metastatic breast cancer, ductal carcinoma in situ, invasive ductal carcinoma, tubular carcinoma, medullary carcinoma, mucinous carcinoma, lobular carcinoma in situ, triple-negative breast cancer; urogenital tract : Kidney (adenocarcinoma, Wilms tumor [neoblastoma], lymphoma, leukemia, renal cell carcinoma, metastatic renal cell carcinoma), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma, urothelial carcinoma), prostate (adenocarcinoma, sarcoma, castration-resistant prostate cancer, bone metastasis, castration-resistant prostate cancer-associated bone metastasis), testis (seminomatosis, teratoma, embryonic carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial carcinoma, fibroma, fibroadenoma, mammaryomatous tumor, lipoma), clear cell carcinoma, papillary carcinoma, penile cancer, penile squamous cell carcinoma; liver : Liver cancer (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; bone : Osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing sarcoma, malignant lymphoma (reticular sarcoma), multiple myeloma, malignant giant cell tumor chordoma, osteochondroma (osteochondrotic exostoma), benign chondroma, chondroblastoma, chondromyxoid osteofibroma, osteoid osteoma and giant cell tumor; thyroid: Medullary thyroid cancer, differentiated thyroid cancer, papillary thyroid cancer, follicular thyroid cancer, Hurstle cell carcinoma, and anaplastic thyroid cancer; nervous system: Skull (osteomatoma, hemangioma, granuloma, xanthomatoma, degenerative osteitis), meninges (meningioma, meningeal sarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germ cell tumor [pineal tumor], glioblastoma pleomorphic, oligodendrocyte, schwannoma, retinoblastoma, congenital tumor), spinal neurofibroma, meningioma, glioma, sarcoma), NF1, neurofibromatosis, plexiform neurofibroma; Gynecology : Uterus (endometrial cancer), cervix (cervical carcinoma, pre-tumor cervical dysplasia), ovary (ovarian carcinoma [serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma], granulosarcous cell tumor, Sertoli-Leydig cell tumor, undifferentiated germ cell tumor, malignant teratoma), vulva (squamous cell carcinoma, carcinoma in situ, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, staphylococcal sarcoma [embryonic rhabdomyosarcoma]), fallopian tube (carcinoma); blood : Blood (myeloid leukemia [acute and chronic], acute lymphoblastic leukemia, chronic lymphoblastic leukemia, myeloproliferative disorders, multiple myeloma, myelodysplastic syndrome), myelofibrosis, polycythemia vera, essential thrombocythemia, Hodgkin's disease, non-Hodgkin lymphoma [malignant lymphoma]; skin : Malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, warts, dysplastic nevus, lipoma, hemangioma, dermatofibroma, keloid, psoriasis; and check : Neuroblastoma. Accordingly, the term “cancerous cell” as provided herein includes a cell suffering from any of the pathological conditions identified above. In some embodiments, compounds or combinations as disclosed herein may be used to treat diseases including HIV, sickle cell disease, graft-versus-host disease, acute graft-versus-host disease, chronic graft-versus-host disease, and sickle cell anemia.

[0076] As defined by the National Cancer Institute, the term "solid tumor" generally refers to a mass of abnormal tissue that does not contain cysts or fluid-filled regions. Solid tumors can be benign (not cancerous) or malignant (cancer). Different types of solid tumors are named according to the type of cells that form them. Examples of solid tumors are sarcomas, carcinomas, and lymphomas. Leukemia (a cancer of the blood) generally does not form solid tumors.

[0077] As defined by the National Cancer Institute, an "immunomodulator" is a substance that stimulates or suppresses the immune system.

[0078] The term “treating” or “treating” refers to any indication of success or improvement in the progression, severity, and / or duration of a disease, pathology, or condition, including any objective or subjective parameters such as decline; remission; reduction of symptoms or making the impairment, pathology, or condition more tolerable to the patient; slowing the rate of degeneration or decline; making the patient less debilitated at the final stage of degeneration; or improving the patient’s physical or mental well-being.

[0079] The term "enhance" refers to an increase or improvement in the function or activity of a protein or cell after the administration of the combination described herein or contact with it, compared to the protein or cell prior to such administration or contact.

[0080] The term “administering” refers to the act of delivering the combinations or compositions described herein to a subject by routes such as oral, mucosal, topical, suppository, intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, or subcutaneous administration. Parenteral administration includes intravenous, intramuscular, intra-arterial, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial administration. Administration generally occurs after the onset of a disease, disorder, or condition or its symptoms, but in certain cases, it may occur before the onset of a disease, disorder, or condition or its symptoms (e.g., administration to a patient predisposed to such disease, disorder, or condition).

[0081] The term “concurrent administration” refers to the administration of two or more agents (e.g., combinations described herein and other active agents, such as anticancer agents described herein). The timing of concurrent administration depends in part on the combinations and compositions administered and may include administration immediately before or immediately after administration, or simultaneously with the administration of one or more additional therapies, such as cancer therapies like chemotherapy, hormone therapy, radiation therapy, or immunotherapy. The compounds of the present invention may be administered to a patient alone or concurrently. Concurrent administration means administering compounds individually or in combination (one or more compounds or agents) simultaneously or sequentially. Thus, formulations may also be combined with other active substances when desired (e.g., to reduce metabolic degradation). The compounds described herein may be used in combination with other active agents known to be useful for cancer treatment.

[0082] The term "anticancer agent" is used in its ordinary, general sense and refers to a composition having anti-neoplastic properties or the ability to inhibit the growth or proliferation of cells. In an embodiment, the anticancer agent is a chemotherapy agent. In an embodiment, the anticancer agent is an agent identified herein that has utility in a method for treating cancer. In an embodiment, the anticancer agent is an agent approved by the FDA or a similar regulatory agency in a country other than the United States for treating cancer.

[0083] The term "chemotherapeutic agent" is used in its ordinary, general sense and refers to a chemical composition or compound having anti-neoplastic properties or the ability to inhibit the growth or proliferation of cells. "Chemotherapy" refers to a regimen or method of care involving the administration of a chemotherapeutic agent or anticancer agent as described herein.

[0084] In general, the nomenclature used in this application is based on the naming rules adopted by the International Union of Pure and Applied Chemistry (IUPAC). The chemical structures presented herein were prepared using CHEMDRAW®. All disclosed valencies appearing on carbon, oxygen, or nitrogen atoms in the structures presented herein indicate the presence of hydrogen atoms.

[0085] Aspects and embodiments

[0086] In one embodiment, the present invention comprises a method for treating cancer in a subject, wherein the method comprises a therapeutically effective amount of compound 1:

[0087]

[0088] or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient,

[0089] The method includes the step of administering a therapeutically effective amount of a gate inhibitor or a pharmaceutical composition containing a gate inhibitor to a subject requiring such treatment.

[0090] In one embodiment, the present invention comprises a method for treating cancer in a subject, wherein the method comprises a compound 1 in a dose of about 5 mg to about 100 mg:

[0091]

[0092] or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient,

[0093] The method includes the step of administering a therapeutically effective amount of a gate inhibitor or a pharmaceutical composition containing a gate inhibitor to a subject requiring such treatment.

[0094] In another aspect, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0095] (i) a step of administering a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof to a subject, wherein compound 1 has the following structure:

[0096] , and

[0097] (ii) A step of administering a therapeutically effective amount of a checkpoint inhibitor to the subject.

[0098] In another aspect, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0099] (i) a step of administering to a subject a dose of about 5 mg to about 100 mg of compound 1 or a pharmaceutically acceptable salt thereof, wherein compound 1 has the following structure: said administering step:

[0100] , and

[0101] (ii) A step of administering a therapeutically effective amount of a checkpoint inhibitor to the subject.

[0102] In another aspect, the present invention comprises a method for treating cancer in a subject, the method comprising the step of administering to a subject requiring such treatment a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound (1) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

[0103] In another aspect, the present invention comprises a method for treating cancer in a subject, the method comprising the step of administering to a subject requiring such treatment a pharmaceutical composition comprising a compound 1 or a pharmaceutically acceptable salt thereof in a dose of about 5 mg to about 100 mg, or a compound (1) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

[0104] In one embodiment, the present invention comprises a method for treating urothelial carcinoma in a subject, the method comprising:

[0105] (i) a step of administering to a subject a dose of about 5 mg to about 100 mg of compound 1 or a pharmaceutically acceptable salt thereof, wherein compound 1 has the following structure: said administering step:

[0106] , and

[0107] (ii) A step of administering a therapeutically effective amount of a checkpoint inhibitor to the subject.

[0108] In another aspect, the present invention comprises a method for treating urothelial carcinoma in a subject, wherein the method comprises a compound 1 in a dose of about 5 mg to about 100 mg:

[0109]

[0110] or a pharmaceutical composition comprising a pharmaceutically acceptable salt or compound (1) or a pharmaceutically acceptable salt and a pharmaceutically acceptable carrier or excipient,

[0111] The method includes the step of administering a therapeutically effective amount of a gate inhibitor or a pharmaceutical composition containing a gate inhibitor to a subject requiring such treatment.

[0112] In one embodiment, the checkpoint inhibitor is selected from PD-1 inhibitors, PD-L1 inhibitors, and CTLA-4 inhibitors.

[0113] In one embodiment, the checkpoint inhibitor is selected from αPD-1 inhibitors, PD-L1 inhibitors, and αCTLA-4 inhibitors.

[0114] In another embodiment, the checkpoint inhibitor is selected from the group consisting of pembrolizumab, nivolumab, atezolizumab, durvalumab, avelumab, cemiplimab, camrelizumab, syntilimab, tisraelizumab, toripalizumab, spartalizumab, dostalimab, KN035 (Jiangshu Alphamab Biopharmaceuticals Company), cocibelimab (formerly CK-301), CA-170 (Curis, Inc.), BMS-986189 (Bristol Myers Squibb Company), and ipilimumab.

[0115] In one embodiment, the checkpoint inhibitor is selected from the group consisting of atezolizumab, avelumab, and nivolumab.

[0116] In one embodiment, the checkpoint inhibitor is pembrolizumab.

[0117] In one embodiment, the checkpoint inhibitor is nivolumab.

[0118] In one embodiment, the checkpoint inhibitor is atezolizumab.

[0119] In one embodiment, the checkpoint inhibitor is avelumab.

[0120] In one embodiment, the checkpoint inhibitor is semiplimab.

[0121] In one embodiment, the checkpoint inhibitor is camrelizumab.

[0122] In one embodiment, the checkpoint inhibitor is syntilimab.

[0123] In one embodiment, the checkpoint inhibitor is tisraelizumab.

[0124] In one embodiment, the checkpoint inhibitor is toripalimab.

[0125] In one embodiment, the checkpoint inhibitor is spartalizumab.

[0126] In one embodiment, the checkpoint inhibitor is dostalimab.

[0127] In one embodiment, the gate inhibitor is KN035.

[0128] In one embodiment, the checkpoint inhibitor is cosivelimab.

[0129] In one embodiment, the gate inhibitor is CA-170.

[0130] In one embodiment, the gate inhibitor is BMS-986189.

[0131] In one embodiment, the checkpoint inhibitor is ipilimumab.

[0132] In one embodiment of the aforementioned aspect, compound 1 or a pharmaceutically acceptable salt thereof is administered orally once a day (qd) or twice a day (bid). In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof is administered orally once a day (qd). In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof is administered orally twice a day (bid).

[0133] In this specification, the dosage of compound 1 is expressed in free base equivalents (FBE) unless otherwise noted.

[0134] In some embodiments, the therapeutically effective amount of compound 1 is about 1 mg to about 500 mg, about 1 mg to about 300 mg, about 1 mg to about 200 mg, about 1 mg to about 150 mg, about 5 mg to about 150 mg, or about 5 mg to about 100 mg.

[0135] In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is about 5 mg to about 80 mg. In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is about 5 mg to about 50 mg.

[0136] In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is 8 mg to 12 mg. In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is 18 mg to 22 mg. In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is 38 mg to 40 mg. In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is about 10 mg. In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is about 20 mg. In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is about 40 mg. In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is about 60 mg. In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is about 80 mg. In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is about 100 mg. In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is about 120 mg. In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is about 140 mg. In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is selected from about 10 mg, 20 mg, and 40 mg. In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is selected from about 10 mg, 20 mg, 40 mg, 60 mg, and 80 mg. In one embodiment, the dosage of compound 1 or a pharmaceutically acceptable salt thereof is selected from about 10 mg, 20 mg, 40 mg, 60 mg, 80 mg, 100 mg, 120 mg, and 140 mg.In one embodiment, a dosage of compound 1 or a pharmaceutically acceptable salt thereof is selected from about 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, and 300 mg.

[0137] In additional embodiments:

[0138] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 100 mg;

[0139] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 95 mg;

[0140] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 90 mg;

[0141] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 85 mg;

[0142] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 80 mg;

[0143] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 75 mg;

[0144] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 70 mg;

[0145] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 65 mg;

[0146] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 60 mg;

[0147] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 55 mg;

[0148] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 50 mg;

[0149] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 45 mg;

[0150] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 40 mg;

[0151] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 35 mg;

[0152] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 30 mg;

[0153] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 25 mg;

[0154] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 20 mg;

[0155] When compound 1 is administered in an amount greater than 0 mg and less than or equal to 15 mg;

[0156] Compound 1 is administered in an amount greater than 0 mg and less than or equal to 10 mg; or

[0157] Compound 1 is administered in an amount greater than 0 mg and less than or equal to 5 mg.

[0158] Solid form of Compound 1

[0159] In the aforementioned aspects and embodiments, compound 1 may be administered in the form of a crystalline (free base) solid or as a crystalline salt.

[0160] Crystalline (free base) solid of Compound 1

[0161] In one embodiment, compound 1 is administered as a crystalline (free base) solid. In one embodiment, the crystalline solid form of compound 1 is characterized as form A, form B, form C, form D, form E, form F, form G, form H, form I, form J, form K, form L, form M, form N, form O, form P, or form Q. In another embodiment, the crystalline solid form of compound 1 is characterized as form A, form B, form C, form D, form E, form F, form G, form H, form K, form O, or form Q. In yet another embodiment, the crystalline solid form of compound 1 is characterized as form I, form J, form L, form M, form N, or form P. Crystalline solid forms of Compound 1 characterized by form A, form B, form C, form D, form E, form F, form G, form H, form I, form J, form K, form L, form M, form N, form O, form P, or form Q are disclosed in WO 2020 / 123800, the entire contents of which are incorporated herein by reference for all purposes.

[0162] In one embodiment, the crystalline solid is characterized as compound 1 form A.

[0163] In one embodiment, the crystalline solid is characterized as compound 1 form B.

[0164] In one embodiment, the crystalline solid is characterized as compound 1 form C.

[0165] In one embodiment, the crystalline solid is characterized as compound 1 form D.

[0166] In one embodiment, the crystalline solid is characterized as compound 1 form E.

[0167] In one embodiment, the crystalline solid is characterized as compound 1 form F.

[0168] In one embodiment, the crystalline solid is characterized as compound 1 form G.

[0169] In one embodiment, the crystalline solid is characterized as compound 1 form H.

[0170] In one embodiment, the crystalline solid is characterized as compound 1 type I.

[0171] In one embodiment, the crystalline solid is characterized as compound 1 form J.

[0172] In one embodiment, the crystalline solid is characterized as compound 1 form K.

[0173] In one embodiment, the crystalline solid is characterized as compound 1 form L.

[0174] In one embodiment, the crystalline solid is characterized as compound 1 form M.

[0175] In one embodiment, the crystalline solid is characterized as compound 1 form N.

[0176] In one embodiment, the crystalline solid is characterized as compound 1 form O.

[0177] In one embodiment, the crystalline solid is characterized as compound 1 form P.

[0178] In one embodiment, the crystalline solid is characterized as compound 1 form Q.

[0179] Crystalline salt of Compound 1

[0180] In another embodiment, compound 1 is administered as a crystalline salt or a hydrate or solvate thereof.

[0181] In one embodiment, the crystalline salt is characterized as compound 1 HCl form A, compound 1 HCl form B, compound 1 HCl form C, or compound 1 HCl form D. The crystalline salt forms characterized as compound 1 HCl form A, compound 1 HCl form B, compound 1 HCl form C, or compound 1 HCl form D are disclosed in WO 2020 / 123800, the entire contents of which are incorporated herein by reference for all purposes.

[0182] In one embodiment, the crystalline salt is characterized as compound 1 HCl form A.

[0183] In one embodiment, the crystalline salt is characterized as compound 1 HCl form B.

[0184] In one embodiment, the crystalline salt is characterized as compound 1 HCl form C.

[0185] In one embodiment, the crystalline salt is characterized as compound 1 HCl form D.

[0186] In one embodiment, the pharmaceutical composition as disclosed herein comprises a crystalline fumarate of Compound 1 or a hydrate or solvate thereof. In some embodiments, the crystalline fumarate of Compound 1 is characterized as Compound 1 fumarate form A or Compound 1 hemifumarate form B. The crystalline fumarate of Compound 1 characterized as Compound 1 fumarate form A or Compound 1 hemifumarate form B is disclosed in WO 2020 / 123800, the entire contents of which are incorporated herein by reference for all purposes.

[0187] In one embodiment, the crystalline salt is characterized as compound 1 fumarate form A.

[0188] In one embodiment, the crystalline fumarate salt is characterized as compound 1 hemifumarate form B.

[0189] Pharmaceutical composition of Compound 1

[0190] In the aforementioned aspects and embodiments, Compound 1 may be administered as a pharmaceutical composition. In one embodiment, the pharmaceutical composition comprises Compound 1 in the form of a crystalline (free base) solid. In another embodiment, the pharmaceutical composition comprises Compound 1 as a crystalline salt.

[0191] In an additional embodiment, the pharmaceutical composition is a tablet.

[0192] In additional embodiments, the tablet pharmaceutical composition comprises the following:

[0193] a. About 20% by weight to about 40% by weight of Compound 1, which is a crystalline solid or a crystalline salt selected from the group consisting of Compound 1 HCl salt, Compound 1 fumarate, and Compound 1 phosphate;

[0194] b. About 35% by weight to about 45% by weight of microcrystalline cellulose;

[0195] c. About 15% by weight to about 25% by weight of lactose;

[0196] d. About 2% by weight to about 8% by weight of hydroxypropyl cellulose;

[0197] e. About 4% to about 8% by weight of croscarmellose sodium;

[0198] f. About 0.1 wt% to about 0.5 wt% of silicon dioxide; and

[0199] g. About 0.5 wt% to about 3.5 wt% of magnesium stearate; and optionally,

[0200] h. Film coating agent.

[0201] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0202] a. About 20% by weight to about 40% by weight of Compound 1, which is a crystalline solid or a crystalline salt selected from the group consisting of Compound 1 HCl salt, Compound 1 fumarate, and Compound 1 phosphate;

[0203] b. About 35% by weight to about 45% by weight of microcrystalline cellulose;

[0204] c. About 15% by weight to about 25% by weight of anhydrous lactose;

[0205] d. About 2% by weight to about 8% by weight of hydroxypropyl cellulose;

[0206] e. About 4% to about 8% by weight of croscarmellose sodium;

[0207] f. About 0.1% by weight to about 0.5% by weight of colloidal silicon dioxide; and

[0208] g. About 0.5 wt% to about 3.5 wt% of magnesium stearate; and optionally,

[0209] h. Film coating agent.

[0210] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0211] a. About 25% by weight to about 35% by weight of Compound 1, which is a crystalline solid or a crystalline salt selected from the group consisting of Compound 1 HCl salt, Compound 1 fumarate, and Compound 1 phosphate;

[0212] b. About 37% by weight to about 43% by weight of microcrystalline cellulose;

[0213] c. About 18% by weight to about 22% by weight of anhydrous lactose;

[0214] d. About 2% by weight to about 6% by weight of hydroxypropyl cellulose;

[0215] e. About 5% by weight to about 7% by weight of croscarmellose sodium;

[0216] f. About 0.2 wt% to about 0.4 wt% of colloidal silicon dioxide; and

[0217] g. About 0.5 wt% to about 3.5 wt% of magnesium stearate; and optionally,

[0218] h. Film coating agent.

[0219] Accordingly, in another embodiment, the tablet pharmaceutical composition comprises the following:

[0220] a. About 20% by weight to about 40% by weight of Compound 1, which is a crystalline solid or a crystalline salt selected from the group consisting of Compound 1 HCl salt, Compound 1 fumarate, and Compound 1 phosphate;

[0221] b. About 35% by weight to about 45% by weight of microcrystalline cellulose;

[0222] c. About 15% by weight to about 25% by weight of lactose;

[0223] d. About 2% by weight to about 8% by weight of hydroxypropyl cellulose;

[0224] e. About 2% by weight to about 8% by weight of croscarmellose sodium;

[0225] f. About 0.1 wt% to about 0.5 wt% of silicon dioxide; and

[0226] g. About 1% by weight to about 5% by weight of stearic acid; and optionally,

[0227] h. Film coating agent.

[0228] Accordingly, in another embodiment, the tablet pharmaceutical composition comprises the following:

[0229] a. About 20% by weight to about 40% by weight of Compound 1, which is a crystalline solid or a crystalline salt selected from the group consisting of Compound 1 HCl salt, Compound 1 fumarate, and Compound 1 phosphate;

[0230] b. About 35% by weight to about 45% by weight of microcrystalline cellulose;

[0231] c. About 15% by weight to about 25% by weight of anhydrous lactose;

[0232] d. About 2% by weight to about 8% by weight of hydroxypropyl cellulose;

[0233] e. About 2% by weight to about 8% by weight of croscarmellose sodium;

[0234] f. About 0.1% by weight to about 0.5% by weight of colloidal silicon dioxide; and

[0235] g. About 1% by weight to about 5% by weight of stearic acid; and optionally,

[0236] h. Film coating agent.

[0237] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0238] a. About 25% by weight to about 35% by weight of Compound 1, which is a crystalline solid or a crystalline salt selected from the group consisting of Compound 1 HCl salt, Compound 1 fumarate, and Compound 1 phosphate;

[0239] b. About 35% by weight to about 40% by weight of microcrystalline cellulose;

[0240] c. About 16% by weight to about 22% by weight of anhydrous lactose;

[0241] d. About 3% by weight to about 7% by weight of hydroxypropyl cellulose;

[0242] e. About 3% by weight to about 7% by weight of croscarmellose sodium

[0243] f. About 0.1% by weight to about 0.5% by weight of colloidal silicon dioxide; and

[0244] g. About 0.5% by weight to about 3.5% by weight of stearic acid; and optionally,

[0245] h. Film coating agent.

[0246] In one embodiment, the pharmaceutical composition of the present disclosure comprises compound 1 as a crystalline (free base) solid.

[0247] In one embodiment, the crystalline solid form of compound 1 is characterized as form A, form B, form C, form D, form E, form F, form G, form H, form I, form J, form K, form L, form M, form N, form O, form P, or form Q. In another embodiment, the crystalline solid form of compound 1 is characterized as form A, form B, form C, form D, form E, form F, form G, form H, form K, form O, or form Q. In yet another embodiment, the crystalline solid form of compound 1 is characterized as form I, form J, form L, form M, form N, or form P. Crystalline solid forms of Compound 1 characterized by form A, form B, form C, form D, form E, form F, form G, form H, form I, form J, form K, form L, form M, form N, form O, form P, or form Q are disclosed in WO 2020 / 123800, the entire contents of which are incorporated herein by reference for all purposes.

[0248] In one embodiment, the crystalline solid is characterized as compound 1 form A.

[0249] In one embodiment, the crystalline solid is characterized as compound 1 form B.

[0250] In one embodiment, the crystalline solid is characterized as compound 1 form C.

[0251] In one embodiment, the crystalline solid is characterized as compound 1 form D.

[0252] In one embodiment, the crystalline solid is characterized as compound 1 form E.

[0253] In one embodiment, the crystalline solid is characterized as compound 1 form F.

[0254] In one embodiment, the crystalline solid is characterized as compound 1 form G.

[0255] In one embodiment, the crystalline solid is characterized as compound 1 form H.

[0256] In one embodiment, the crystalline solid is characterized as compound 1 type I.

[0257] In one embodiment, the crystalline solid is characterized as compound 1 form J.

[0258] In one embodiment, the crystalline solid is characterized as compound 1 form K.

[0259] In one embodiment, the crystalline solid is characterized as compound 1 form L.

[0260] In one embodiment, the crystalline solid is characterized as compound 1 form M.

[0261] In one embodiment, the crystalline solid is characterized as compound 1 form N.

[0262] In one embodiment, the crystalline solid is characterized as compound 1 form O.

[0263] In one embodiment, the crystalline solid is characterized as compound 1 form P.

[0264] In one embodiment, the crystalline solid is characterized as compound 1 form Q.

[0265] In another embodiment, the pharmaceutical composition of the present disclosure comprises compound 1 as a crystalline salt or a hydrate or solvate thereof.

[0266] In one embodiment, the crystalline salt is characterized as compound 1 HCl form A, compound 1 HCl form B, compound 1 HCl form C, or compound 1 HCl form D. The crystalline salt forms characterized as compound 1 HCl form A, compound 1 HCl form B, compound 1 HCl form C, or compound 1 HCl form D are disclosed in WO 2020 / 123800, the entire contents of which are incorporated herein by reference for all purposes.

[0267] In one embodiment, the crystalline salt is characterized as compound 1 HCl form A.

[0268] In one embodiment, the crystalline salt is characterized as compound 1 HCl form B.

[0269] In one embodiment, the crystalline salt is characterized as compound 1 HCl form C.

[0270] In one embodiment, the crystalline salt is characterized as compound 1 HCl form D.

[0271] In one embodiment, the pharmaceutical composition as disclosed herein comprises a crystalline fumarate of Compound 1 or a hydrate or solvate thereof. In some embodiments, the crystalline fumarate of Compound 1 is characterized as Compound 1 fumarate form A or Compound 1 hemifumarate form B. The crystalline fumarate of Compound 1 characterized as Compound 1 fumarate form A or Compound 1 hemifumarate form B is disclosed in WO 2020 / 123800, the entire contents of which are incorporated herein by reference for all purposes.

[0272] In one embodiment, the crystalline salt is characterized as compound 1 fumarate form A.

[0273] In one embodiment, the crystalline fumarate salt is characterized as compound 1 hemifumarate form B.

[0274] In one embodiment, the pharmaceutical composition comprises a crystalline phosphate of compound 1 or a hydrate or solvate thereof. In some embodiments, the crystalline phosphate of compound 1 is characterized as compound 1 phosphate form A. The crystalline phosphate of compound 1 characterized as compound 1 phosphate form A is disclosed in WO 2020 / 123800, the entire contents of which are incorporated herein by reference for all purposes.

[0275] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0276] a. About 25% by weight to about 35% by weight of compound 1 hemifumarate salt;

[0277] b. About 37% by weight to about 43% by weight of microcrystalline cellulose;

[0278] c. About 18% by weight to about 22% by weight of anhydrous lactose;

[0279] d. About 2% by weight to about 6% by weight of hydroxypropyl cellulose;

[0280] e. About 5% by weight to about 7% by weight of croscarmellose sodium;

[0281] f. About 0.2 wt% to about 0.4 wt% of colloidal silicon dioxide; and

[0282] g. About 0.5 wt% to about 3.5 wt% of magnesium stearate; and optionally,

[0283] h. Film coating agent.

[0284] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0285] a. Approximately 25% by weight to approximately 35% by weight of compound 1 hemifumarate salt form B;

[0286] b. About 37% by weight to about 43% by weight of microcrystalline cellulose;

[0287] c. About 18% by weight to about 22% by weight of anhydrous lactose;

[0288] d. About 2% by weight to about 6% by weight of hydroxypropyl cellulose;

[0289] e. About 5% by weight to about 7% by weight of croscarmellose sodium;

[0290] f. About 0.2 wt% to about 0.4 wt% of colloidal silicon dioxide; and

[0291] g. About 0.5 wt% to about 3.5 wt% of magnesium stearate; and optionally,

[0292] h. Film coating agent.

[0293] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0294] a. Approximately 27.75 wt% of compound 1 hemifumarate salt;

[0295] b. Approximately 41.47 wt% of microcrystalline cellulose;

[0296] c. Approximately 20.73 wt% anhydrous lactose;

[0297] d. About 3 weight percent hydroxypropyl cellulose;

[0298] e. Approximately 6 weight percent of croscarmellose sodium;

[0299] f. Approximately 0.3 wt% of colloidal silicon dioxide; and

[0300] g. About 0.75 wt% of magnesium stearate; and optionally,

[0301] h. Film coating agent.

[0302] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0303] a. Approximately 27.75 wt% of compound 1 hemifumarate salt form B;

[0304] b. Approximately 41.47 wt% of microcrystalline cellulose;

[0305] c. Approximately 20.73 wt% anhydrous lactose;

[0306] d. About 3 weight percent hydroxypropyl cellulose;

[0307] e. Approximately 6 weight percent of croscarmellose sodium;

[0308] f. Approximately 0.3 wt% of colloidal silicon dioxide; and

[0309] g. About 0.75 wt% of magnesium stearate; and optionally,

[0310] h. Film coating agent.

[0311] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0312] a. 20 mg to 25 mg of compound 1 hemifumarate salt;

[0313] b. 30 mg to 35 mg of microcrystalline cellulose;

[0314] c. 15 mg to 18 mg of anhydrous lactose;

[0315] d. 1.5 mg to 4.5 mg of hydroxypropyl cellulose;

[0316] e. 4 mg to 6 mg of croscarmellose sodium;

[0317] f. 0.1 mg to 0.3 mg of colloidal silicon dioxide; and

[0318] g. 0.5 mg to 0.7 mg of magnesium stearate; and optionally,

[0319] h. 2 mg to 6 mg of film coating agent.

[0320] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0321] a. 20 mg to 25 mg of compound 1 hemifumarate salt form B;

[0322] b. 30 mg to 35 mg of microcrystalline cellulose;

[0323] c. 15 mg to 18 mg of anhydrous lactose;

[0324] d. 1.5 mg to 4.5 mg of hydroxypropyl cellulose;

[0325] e. 4 mg to 6 mg of croscarmellose sodium;

[0326] f. 0.1 mg to 0.3 mg of colloidal silicon dioxide; and

[0327] g. 0.5 mg to 0.7 mg of magnesium stearate; and optionally,

[0328] h. 2 mg to 6 mg of film coating agent.

[0329] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0330] a. 22.20 mg of compound 1 hemifumarate salt form B;

[0331] b. 30 mg to 35 mg of microcrystalline cellulose;

[0332] c. 15 mg to 18 mg of anhydrous lactose;

[0333] d. 1.5 mg to 4.5 mg of hydroxypropyl cellulose;

[0334] e. 4 mg to 6 mg of croscarmellose sodium;

[0335] f. 0.1 mg to 0.3 mg of colloidal silicon dioxide; and

[0336] g. 0.5 mg to 0.7 mg of magnesium stearate; and optionally,

[0337] h. 2 mg to 6 mg of film coating agent.

[0338] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0339] a. 22.20 mg of compound 1 hemifumarate salt form B;

[0340] b. 33.17 mg of microcrystalline cellulose;

[0341] c. 16.59 mg of anhydrous lactose;

[0342] d. 2.4 mg of hydroxypropyl cellulose;

[0343] e. 4.8 mg of croscarmellose sodium;

[0344] f. 0.24 mg of colloidal silicon dioxide; and

[0345] g. 0.6 mg of magnesium stearate; and optionally,

[0346] h. 3.2 mg of film coating agent.

[0347] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0348] a. About 25% by weight to about 35% by weight of compound 1 hemifumarate salt;

[0349] b. About 35% by weight to about 40% by weight of microcrystalline cellulose;

[0350] c. About 16% by weight to about 22% by weight of anhydrous lactose;

[0351] d. About 3% by weight to about 7% by weight of hydroxypropyl cellulose;

[0352] e. About 3% by weight to about 7% by weight of croscarmellose sodium

[0353] f. About 0.1% by weight to about 0.5% by weight of colloidal silicon dioxide; and

[0354] g. About 0.5% by weight to about 3.5% by weight of stearic acid; and optionally,

[0355] h. Film coating agent.

[0356] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0357] a. Approximately 25% by weight to approximately 35% by weight of compound 1 hemifumarate salt form B;

[0358] b. About 35% by weight to about 40% by weight of microcrystalline cellulose;

[0359] c. About 16% by weight to about 22% by weight of anhydrous lactose;

[0360] d. About 3% by weight to about 7% by weight of hydroxypropyl cellulose;

[0361] e. About 3% by weight to about 7% by weight of croscarmellose sodium

[0362] f. About 0.1% by weight to about 0.5% by weight of colloidal silicon dioxide; and

[0363] g. About 0.5% by weight to about 3.5% by weight of stearic acid; and optionally,

[0364] h. Film coating agent.

[0365] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0366] a. Approximately 27.75 wt% of compound 1 hemifumarate salt;

[0367] b. Approximately 38.63 wt% of microcrystalline cellulose;

[0368] c. Approximately 19.32 wt% anhydrous lactose;

[0369] d. About 5% by weight of hydroxypropyl cellulose;

[0370] e. Approximately 6 wt% of croscarmellose sodium

[0371] f. Approximately 0.3 wt% of colloidal silicon dioxide; and

[0372] g. About 3% by weight of stearic acid; and optionally,

[0373] h. Film coating agent.

[0374] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0375] a. Approximately 27.75 wt% of compound 1 hemifumarate salt form B;

[0376] b. Approximately 38.63 wt% of microcrystalline cellulose;

[0377] c. Approximately 19.32 wt% anhydrous lactose;

[0378] d. About 5% by weight of hydroxypropyl cellulose;

[0379] e. Approximately 6 wt% of croscarmellose sodium

[0380] f. Approximately 0.3 wt% of colloidal silicon dioxide; and

[0381] g. About 3% by weight of stearic acid; and optionally,

[0382] h. Film coating agent.

[0383] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0384] a. 20 mg to 25 mg of compound 1 hemifumarate salt;

[0385] b. 30 mg to 40 mg of microcrystalline cellulose;

[0386] c. 15 mg to 20 mg of anhydrous lactose;

[0387] d. 3 mg to 7 mg of hydroxypropyl cellulose;

[0388] e. 3 mg to 7 mg of croscarmellose sodium;

[0389] f. 0.1 mg to 0.3 mg of colloidal silicon dioxide; and

[0390] g. 2 mg to 4 mg of stearic acid; and optionally,

[0391] h. 2 mg to 5 mg of film coating agent.

[0392] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0393] a. 20 mg to 25 mg of compound 1 hemifumarate salt form B;

[0394] b. 30 mg to 40 mg of microcrystalline cellulose;

[0395] c. 15 mg to 20 mg of anhydrous lactose;

[0396] d. 3 mg to 7 mg of hydroxypropyl cellulose;

[0397] e. 3 mg to 7 mg of croscarmellose sodium;

[0398] f. 0.1 mg to 0.3 mg of colloidal silicon dioxide; and

[0399] g. 2 mg to 4 mg of stearic acid; and optionally,

[0400] h. 2 mg to 5 mg of film coating agent.

[0401] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0402] a. 22.20 mg of compound 1 hemifumarate salt form B;

[0403] b. 30 mg to 40 mg of microcrystalline cellulose;

[0404] c. 15 mg to 20 mg of anhydrous lactose;

[0405] d. 3 mg to 7 mg of hydroxypropyl cellulose;

[0406] e. 3 mg to 7 mg of croscarmellose sodium;

[0407] f. 0.1 mg to 0.3 mg of colloidal silicon dioxide; and

[0408] g. 2 mg to 4 mg of stearic acid; and optionally,

[0409] h. 2 mg to 5 mg of film coating agent.

[0410] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0411] a. 22.20 mg of compound 1 hemifumarate salt form B;

[0412] b. 30.9 mg of microcrystalline cellulose;

[0413] c. 15.46 mg of anhydrous lactose;

[0414] d. 4 mg of hydroxypropyl cellulose;

[0415] e. 4.8 mg of croscarmellose sodium;

[0416] f. 0.24 mg of colloidal silicon dioxide; and

[0417] g. 2.4 mg of stearic acid; and optionally,

[0418] h. 3.2 mg of film coating agent.

[0419] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0420] a. 83 mg to 93 mg of compound 1 hemifumarate salt;

[0421] b. 120 mg to 150 mg of microcrystalline cellulose;

[0422] c. 60 mg to 80 mg of anhydrous lactose;

[0423] d. 12 mg to 30 mg of hydroxypropyl cellulose;

[0424] e. 12 mg to 30 mg of croscarmellose sodium;

[0425] f. 0.5 mg to 1.5 mg of colloidal silicon dioxide; and

[0426] g. 8 mg to 16 mg of stearic acid; and optionally,

[0427] h. 8 mg to 14 mg of film coating agent.

[0428] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0429] a. 83 mg to 93 mg of compound 1 hemifumarate salt form B;

[0430] b. 120 mg to 150 mg of microcrystalline cellulose;

[0431] c. 60 mg to 80 mg of anhydrous lactose;

[0432] d. 12 mg to 30 mg of hydroxypropyl cellulose;

[0433] e. 12 mg to 30 mg of croscarmellose sodium;

[0434] f. 0.5 mg to 1.5 mg of colloidal silicon dioxide; and

[0435] g. 8 mg to 16 mg of stearic acid; and optionally,

[0436] h. 8 mg to 14 mg of film coating agent.

[0437] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0438] a. 88.78 mg of compound 1 hemifumarate salt form B;

[0439] b. 120 mg to 150 mg of microcrystalline cellulose;

[0440] c. 60 mg to 80 mg of anhydrous lactose;

[0441] d. 12 mg to 30 mg of hydroxypropyl cellulose;

[0442] e. 12 mg to 30 mg of croscarmellose sodium;

[0443] f. 0.5 mg to 1.5 mg of colloidal silicon dioxide; and

[0444] g. 8 mg to 16 mg of stearic acid; and optionally,

[0445] h. 8 mg to 14 mg of film coating agent.

[0446] In one embodiment, the tablet pharmaceutical composition comprises the following:

[0447] a. 88.78 mg of compound 1 hemifumarate salt form B;

[0448] b. 123.62 mg of microcrystalline cellulose;

[0449] c. 61.82 mg of anhydrous lactose;

[0450] d. 16 mg of hydroxypropyl cellulose;

[0451] e. 19.2 mg of croscarmellose sodium;

[0452] f. 0.96 mg of colloidal silicon dioxide; and

[0453] g. 9.6 mg of stearic acid; and optionally,

[0454] h. 12.8 mg of film coating agent.

[0455] In another embodiment, compound 1 is administered as a tablet pharmaceutical composition as provided in the following table.

[0456]

[0457] In another embodiment, compound 1 is administered as a tablet pharmaceutical composition as provided in the following table.

[0458]

[0459] Any formulation provided above may be adjusted according to the desired dosage of compound 1. Accordingly, the amount of each formulation component may be proportionally adjusted to provide tablet formulations containing various amounts of compound 1 as provided in the previous paragraph.

[0460] Cancer treatment

[0461] In the aforementioned aspects and embodiments, compound 1 is administered together with a checkpoint inhibitor and optionally an additional immunomodulator to treat cancer.

[0462] In the aforementioned aspects and embodiments, compound 1 is administered as a single agent to treat cancer.

[0463] In one embodiment, the cancer is selected from heart cancer, head and neck cancer, lung cancer, colon cancer, gastrointestinal cancer, breast cancer, genitourinary cancer, liver cancer, bone cancer, thyroid cancer, nervous system cancer, gynecological cancer, blood cancer, skin cancer, and adrenal cancer.

[0464] In additional embodiments, the heart cancer is selected from angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma, myxoma, rhabdomyosarcoma, fibroma, lipoma, and teratoma.

[0465] In another additional embodiment, the head and neck cancer is selected from squamous cell carcinoma of the head and neck, laryngeal cancer and hypopharyngeal cancer, nasal cavity cancer and paranasal sinus cancer, nasopharyngeal cancer, salivary gland cancer, oral cancer and oropharyngeal cancer.

[0466] In another additional embodiment, the lung cancer is selected from squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma and non-small cell lung cancer; and from bronchial carcinoma selected from alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma and mesothelioma.

[0467] In another additional embodiment, colon cancer is selected from colorectal cancer, adenocarcinoma, gastrointestinal stromal tumor, lymphoma, carcinoid, and Turcot syndrome.

[0468] In another additional embodiment, gastric cancer is selected from gastric cancer, gastroesophageal junction adenocarcinoma, esophageal squamous cell carcinoma, esophageal adenocarcinoma, esophageal leiomyosarcoma, esophageal lymphoma, gastric carcinoma, gastric lymphoma, gastric leiomyosarcoma, pancreatic ductal adenocarcinoma, pancreatic insulinoma, pancreatic glucagonoma, pancreatic gastrinoma, pancreatic carcinoid tumor, biforma, small intestine adenocarcinoma, small intestine lymphoma, small intestine carcinoid tumor, small intestine Kaposi sarcoma, small intestine leiomyoma, small intestine hemangioma, small intestine lipoma, small intestine neurofibroma, small intestine fibroma, colon adenocarcinoma, colon tubular adenoma, colon villous adenoma, colon hamartoma and colon leiomyoma.

[0469] In another additional embodiment, breast cancer is selected from metastatic breast cancer, ductal carcinoma in situ, invasive ductal carcinoma, tubular carcinoma, medullary carcinoma, mucinous carcinoma, lobular carcinoma in situ, and triple-negative breast cancer.

[0470] In another additional embodiment, the genitourinary cancer is selected from renal adenocarcinoma, renal nephroblastoma, renal lymphoma, renal cell carcinoma, squamous cell carcinoma of the bladder or urethra, transitional cell carcinoma of the bladder or urethra, adenocarcinoma of the bladder or urethra, urothelial carcinoma of the bladder or urethra, prostate adenocarcinoma, prostate sarcoma, castration-resistant prostate cancer, seminoma, testicular teratoma, embryonic carcinoma, testicular teratoma, testicular choriocarcinoma, testicular sarcoma, testicular interstitial carcinoma, testicular fibroma, testicular fibroadenoma, testicular mammaryomatous tumor, testicular lipoma, clear cell carcinoma, and papillary carcinoma.

[0471] In another additional embodiment, the liver cancer is selected from hepatocellular carcinoma, cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, and angiomas.

[0472] In another additional embodiment, the bone cancer is selected from osteogenic sarcoma, fibrosarcoma, malignant fibrohistiocytoma, chondrosarcoma, Ewing sarcoma, malignant lymphoma, reticular sarcoma, multiple myeloma, malignant giant cell tumor chordoma, osteochondroma, benign chondroma, chondroblastoma, chondromyxoid osteofibroma, osteoostoma and giant cell tumor.

[0473] In another additional embodiment, the thyroid cancer is selected from medullary thyroid cancer, differentiated thyroid cancer, papillary thyroid cancer, follicular thyroid cancer, Hurstle cell carcinoma, and anaplastic thyroid cancer.

[0474] In another additional embodiment, the nervous system cancer is selected from osteoma of the skull, hemangioma of the skull, granuloma of the skull, xanthomas of the skull, degenerative osteitis of the skull, meningioma, meningeal sarcoma, meningeal gliomatosis, cerebral astrocytoma, medulloblastoma, glioma, cerebral ependymoma, germ cell tumor [pineal tumor], glioblastoma pleomorphic, oligodendrocyte tumor, schwannoma, retinoblastoma, congenital brain tumor, spinal neurofibroma, meningioma, and cerebral sarcoma.

[0475] In another additional embodiment, the gynecological cancer is selected from endometrial cancer, cervical carcinoma, pre-tumor cervical dysplasia, ovarian carcinoma selected from serous cystadenocarcinoma, mucinous cystadenocarcinoma and unclassified ovarian carcinoma, granulosarcoma, Sertoli-Leydig cell tumor, undifferentiated germ cell tumor and malignant teratoma; squamous cell carcinoma of the vulva, carcinoma in situ of the vulva, adenocarcinoma of the vulva, fibrosarcoma of the vulva, melanoma of the vulva, clear cell carcinoma of the vagina, squamous cell carcinoma of the vagina, embryonic rhabdomyosarcoma and fallopian tube carcinoma.

[0476] In another additional embodiment, the blood cancer is selected from myeloid leukemia [acute and chronic], acute lymphoblastic leukemia, chronic lymphoblastic leukemia, myeloproliferative disorders, multiple myeloma, myelodysplastic syndrome), Hodgkin's disease and non-Hodgkin lymphoma [malignant lymphoma].

[0477] In another additional embodiment, the skin cancer is selected from malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, wart dysplastic nevus, lipoma, hemangioma, dermatofibroma, keloid, and psoriasis.

[0478] In one embodiment, the cancer is selected from heart cancer, head and neck cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, non-clear cell renal cell carcinoma, advanced clear cell renal carcinoma, castration-resistant prostate cancer, hormone receptor-positive breast cancer, prostate cancer, colon cancer, gastrointestinal cancer, breast cancer, genitourinary cancer, liver cancer, bone cancer, thyroid cancer, nervous system cancer, gynecological cancer, hematological cancer, skin cancer, and adrenal cancer.

[0479] In an additional embodiment, the cancer is a solid tumor.

[0480] In an additional embodiment, the cancer is a solid tumor that is inoperable or locally advanced, metastatic, or recurrent.

[0481] In additional embodiments, the solid tumor is unresectable or metastatic, and life-prolonging therapy is not available or currently available therapy is intolerable or no longer effective.

[0482] In another additional embodiment, the cancer or solid tumor is ICI-refractory.

[0483] In another additional embodiment, the cancer or solid tumor is platinum-refractory.

[0484] In additional embodiments, the solid tumor is a sarcoma, carcinoma, or lymphoma.

[0485] In an additional embodiment, the cancer is advanced clear cell renal carcinoma, hormone receptor-positive breast cancer, or castration anxiety-resistant prostate cancer.

[0486] In one embodiment, the cancer is an advanced clear cell renal carcinoma.

[0487] In one embodiment, the cancer is an unresectable advanced or metastatic clear cell renal cell carcinoma.

[0488] In one embodiment, the cancer is a non-clear cell renal cell carcinoma.

[0489] In one embodiment, the cancer is an advanced non-clear cell renal cell carcinoma.

[0490] In one embodiment, the cancer is an unresectable advanced or metastatic non-clear cell renal cell carcinoma.

[0491] In one embodiment, unresectable advanced or metastatic non-clear cell renal cell carcinoma includes papillary renal cell carcinoma, unclassified renal cell carcinoma, and sarcoid renal cell carcinoma.

[0492] In one embodiment, the cancer is hormone receptor-positive breast cancer.

[0493] In one embodiment, the cancer is castration anxiety-resistant prostate cancer.

[0494] In an additional embodiment, castration anxiety-resistant prostate cancer is metastatic.

[0495] In another additional embodiment, the adrenal cancer is a neuroblastoma.

[0496] In another additional embodiment, the cancer is a urothelial carcinoma.

[0497] In another additional embodiment, the urothelial carcinoma is a locally advanced or metastatic transitional cell carcinoma of the urothelium.

[0498] In another additional embodiment, the cancer is an advanced urothelial carcinoma.

[0499] In another additional embodiment, the cancer is a metastatic urothelial carcinoma.

[0500] In another additional embodiment, the cancer is an ICI-refractory urothelial carcinoma.

[0501] In another additional embodiment, the cancer is a platinum-refractory urothelial carcinoma.

[0502] In another additional embodiment, the cancer is a urothelial carcinoma of the renal pelvis, ureter, bladder, or urethra.

[0503] In another additional embodiment, the cancer is a urothelial carcinoma of the renal pelvis.

[0504] In another additional embodiment, the cancer is a urothelial carcinoma of the ureter.

[0505] In another additional embodiment, the cancer is a urothelial carcinoma of the urethra.

[0506] In another additional embodiment, the cancer is a urothelial carcinoma of the bladder.

[0507] In another embodiment, the cancer is selected from endometrial cancer, sarcoma, neuroendocrine tumor, ovarian cancer, colorectal cancer, HCC, NSCLC, gastric cancer, and melanoma.

[0508] In another embodiment, the cancer is endometrial cancer.

[0509] In another embodiment, cancer is a sarcoma.

[0510] In another embodiment, the cancer is a neuroendocrine tumor.

[0511] In another embodiment, the cancer is ovarian cancer.

[0512] In another embodiment, the cancer is colorectal cancer.

[0513] In another embodiment, the colorectal cancer is right colorectal cancer (RCRC) or left colorectal cancer (LCRC).

[0514] In another embodiment, the cancer is hepatocellular carcinoma.

[0515] In another embodiment, the cancer is non-small cell lung cancer.

[0516] In another embodiment, the cancer is stomach cancer.

[0517] In another embodiment, the cancer is melanoma.

[0518] In another embodiment, the cancer is a solid tumor. In an additional embodiment, the cancer is an unresectable advanced or metastatic solid tumor. In an additional embodiment, the solid tumor is a genitourinary cancer. In an additional embodiment, the genitourinary cancer is selected from the group consisting of clear cell renal cell carcinoma (ccRCC), non-clear cell renal cell carcinoma (nccRCC), urothelial carcinoma (UC, ICI inexperienced and experienced), and metastatic castration-resistant prostate cancer (mCRPC).

[0519] In one embodiment, the subject is a human.

[0520] In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1, and a checkpoint inhibitor or a pharmaceutical composition comprising a checkpoint inhibitor are administered simultaneously, sequentially, or individually.

[0521] In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1, and a pharmaceutical composition comprising a checkpoint inhibitor or an immunomodulator are administered simultaneously, sequentially, or individually. In an embodiment, the immunomodulator is selected from the group consisting of PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, and IL-2 targeting agents.

[0522] In one embodiment, the method further comprises the step of evaluating treatment using the combination therapy by determining one or more of the following: inhibition of disease progression, inhibition of tumor growth, reduction of primary tumor, alleviation of tumor-related symptoms, inhibition of tumor-secreted factors, delay in the appearance of primary or secondary tumors, slowing of the development of primary or secondary tumors, reduction in the occurrence of primary or secondary tumors, slowing of secondary effects of disease or reduction in severity, arrest of tumor growth and regression of tumors, increase in Time To Progression (TTP), increase in Progression Free Survival (PFS), increase in overall response rate, increase in Overall Survival (OS) or increase in Duration of Response (DOR), and change in tumor markers from baseline.

[0523] In one embodiment, the method comprises the treatment of cancer that has not previously been treated with any other anticancer treatment. In another embodiment, the method comprises the treatment of cancer that has not previously been treated with a PD-1 inhibitor. In another embodiment, the method comprises the treatment of cancer that has not previously been treated with a PD-1 inhibitor. In another embodiment, the method comprises the treatment of cancer that has previously been treated with pembrolizumab, nivolumab, atezolizumab, durvalumab, avelumab, cemiflimab, camrelizumab, syntilimab, tisreilizumab, toripalimab, spartalizumab, dostalimab, KN035, cocibelimab, CA-170 (Curis, Inc.), or BMS-986189. In another embodiment, the method comprises the treatment of cancer that has not been treated with pembrolizumab, nivolumab, atezolizumab, durvalumab, avelumab, cemiplimab, camrelizumab, syntilimab, tisreilizumab, toripalimab, spartalizumab, dostalimab, KN035, cocibelimab, CA-170 (Curis, Inc.), or BMS-986189.

[0524] In one embodiment, the method comprises treating cancer that has previously been treated with a PD-1 inhibitor, wherein the treatment initially showed a partial response but later developed resistance to PD-1 with disease progression.

[0525] In one embodiment, the method comprises treating cancer that has previously been treated with a PD-1 inhibitor, wherein the treatment initially showed stable lesions but later developed resistance to PD-1 with disease progression.

[0526] In one embodiment, the method comprises treating cancer that has previously been treated with a PD-1 inhibitor, wherein the treatment initially showed a complete response but later developed resistance to PD-1 with disease progression.

[0527] In one embodiment, the method comprises treating cancer that has previously been treated with a PD-1 inhibitor, but the treatment did not show a response to treatment.

[0528] In one embodiment, the method comprises treating cancer that has never been previously treated with a PD-1 inhibitor, wherein the treatment initially showed a partial response but later developed resistance to PD-1 with disease progression.

[0529] In one embodiment, the method comprises treating cancer that has not previously been treated with a PD-1 inhibitor, wherein the treatment initially showed stable lesions but later developed resistance to PD-1 with disease progression.

[0530] In one embodiment, the method comprises treating cancer that has never been previously treated with a PD-1 inhibitor, wherein the treatment initially showed a complete response but later developed resistance to PD-1 with disease progression.

[0531] In one embodiment, the method comprises treating cancer that has not previously been treated with a PD-1 inhibitor, but the treatment did not show a response to treatment.

[0532] In embodiments of the present disclosure, a checkpoint inhibitor or PD-1 inhibitor combined with a non-polymorphic form, crystalline form, or crystalline salt form of compound 1 is used to inhibit or reduce tumor or cancer recurrence or tumor or cancer progression by reducing or inhibiting metastasis of the primary tumor or cancer to another site, or the formation or establishment of a metastatic tumor or cancer in another site away from the primary tumor or cancer.

[0533] In additional embodiments of the present disclosure, a combination therapy for treating cancer is provided herein, comprising a non-polymorphic form, a crystalline form or a crystalline salt form of compound 1 and a checkpoint inhibitor or a PD-1 inhibitor, which has the potential to induce a potent and sustained immune response with enhanced therapeutic benefits and more manageable toxicity.

[0534] In additional embodiments of the present disclosure, a combination therapy for treating cancer comprising a non-polymorphic form, a crystalline form, or a crystalline salt form of Compound 1 and a checkpoint inhibitor or a PD-1 inhibitor is provided herein. In embodiments of the present disclosure, a method for treating cancer and / or preventing the establishment of metastasis by using a non-polymorphic form, a crystalline form, or a crystalline salt form of Compound 1 of the present invention that acts synergistically with a checkpoint inhibitor is provided herein.

[0535] In additional embodiments, the present disclosure provides a method for one or more of the following: 1) reducing or inhibiting the growth, proliferation, motility, or invasiveness of a tumor or cancer cell that is metastatic or causes metastasis; 2) reducing or inhibiting the formation or establishment of a metastasis arising from the primary tumor or cancer in one or more other sites, locations, or regions distinct from the primary tumor or cancer; 3) reducing or inhibiting the growth or proliferation of a metastasis in one or more other sites, locations, or regions distinct from the primary tumor or cancer after the metastasis has formed or established; 4) reducing or inhibiting the formation or establishment of additional metastases after the metastasis has formed or established; 5) extended overall survival; 6) extended progression-free survival; or 7) disease stabilization. The method comprises the step of administering a non-polymorphic form, a crystalline form, or a crystalline salt form of Compound 1 of the present invention in combination with a checkpoint inhibitor as described herein to a subject in need thereof.

[0536] In embodiments of the present disclosure, administration of a non-polymorphic form, crystalline form, or crystalline salt form of compound 1 combined with a checkpoint inhibitor or PD-1 inhibitor provides a detectable or measurable improvement, i.e., a therapeutic benefit or beneficial effect, in a given subject's pathological condition, such as alleviating or improving one or more adverse (physical) symptoms or outcomes associated with the presence of a proliferative or hyperproliferative disorder, a neoplasm, a tumor or cancer, or metastasis.

[0537] Therapeutic benefit or beneficial effect is any objective or subjective, transient or long-term improvement of a condition or pathology, or a reduction in the onset, severity, duration, or frequency of adverse symptoms associated with or caused by a proliferative or hyperproliferative disorder such as a neoplasm, tumor, or cancer, or metastasis. This may lead to an improvement in survival rate. A satisfactory clinical endpoint of the treatment method according to the present disclosure is achieved, for example, when there is a progressive or partial reduction in the severity, duration, or frequency of one or more associated pathologies, adverse symptoms, or complications, or when there is inhibition or reversal of one or more physiological, biochemical, or cellular signs or characteristics of a proliferative or hyperproliferative disorder such as a neoplasm, tumor, or cancer, or metastasis. Therapeutic benefit or improvement may be, but is not limited to, the destruction of target proliferating cells (e.g., neoplasms, tumors, or cancers, or metastases) or the elimination of one or more, most or all, pathological adverse symptoms or complications associated with or caused by proliferative or hyperproliferative disorders such as neoplasms, tumors, or cancers, or metastases. However, therapeutic benefit or improvement does not necessarily have to be the healing or complete destruction of all target proliferating cells (e.g., neoplasms, tumors, or cancers, or metastases) or the elimination of all pathological adverse symptoms or complications associated with or caused by proliferative or hyperproliferative disorders such as neoplasms, tumors, or cancers, or metastases. For example, the partial destruction of a tumor or cancer cell mass, or the stabilization of the tumor or cancer mass, size, or number of cells by inhibiting the progression or deterioration of the tumor or cancer, may reduce mortality and extend lifespan for only a few days, weeks, or months, even if some or most of the tumor or cancer mass, size, or cells remain.

[0538] Specific, non-limiting examples of therapeutic benefits include a reduction in the volume (size or cell mass) or number of cells of a neoplasm, tumor, or cancer, or metastasis; inhibition or prevention of the increase in the volume of a neoplasm, tumor, or cancer (e.g., stabilization); slowing or inhibition of the progression, deterioration, or metastasis of a neoplasm, tumor, or cancer; or inhibition of the proliferation, growth, or metastasis of a neoplasm, tumor, or cancer.

[0539] In embodiments of the present disclosure, administration of a checkpoint inhibitor or PD-1 inhibitor in combination therapy with the non-polymorphic form, crystalline form, or crystalline salt form of Compound 1 provides a detectable or measurable improvement or overall response rate according to irRC (derived from time-in-time response evaluation and based on tumor burden), including one or more of the following: (i) irCR -- complete resolution of all lesions and no new lesions, regardless of measurability (confirmed by continuous evaluation repeated for at least 4 weeks from the date of first documenting), (ii) irPR -- reduction of tumor burden by 50% or more compared to baseline (confirmed by continuous evaluation for at least 4 weeks after first documenting).

[0540] Optionally, any method described herein may not produce immediate effects. For example, an increase in the number or mass of neoplasms, tumors, or cancer cells may follow treatment, but over time, ultimate stabilization or reduction of the tumor cell mass, size, or number of cells in a given subject may subsequently occur.

[0541] Additional adverse symptoms and complications associated with neoplasms, tumors, cancers, and metastases that can be inhibited, reduced, delayed, or prevented include, for example, nausea, loss of appetite, lethargy, pain, and discomfort. Thus, a partial or complete decrease or reduction in the severity, duration, or frequency of adverse symptoms or complications associated with or caused by cellular hyperproliferative disorders, improvement in the subject's quality of life, and / or well-being such as increased energy, appetite, and mental well-being are all specific, non-limiting examples of therapeutic benefits.

[0542] Accordingly, therapeutic benefits or improvements may also include a subjective improvement in the quality of life of the subject being treated. In additional embodiments, the method prolongs or extends the lifespan (survival) of the subject. In additional embodiments, the method improves the quality of life of the subject.

[0543] In one embodiment, administration of a checkpoint inhibitor or PD-1 inhibitor in combination therapy with the non-polymorphic form, crystalline form, or crystalline salt form of compound 1 results in clinically relevant improvement in one or more markers of disease status and progression selected from one or more of the following: (i) overall survival, (ii) progression-free survival, (iii) overall response rate, (iv) reduction in metastatic disease, (v) circulating levels of tumor antigens such as carbohydrate antigen 19.9 (CA19.9) and carcinembryonic antigen (CEA) or other tumor-dependent antigens, (vii) nutritional status (body weight, appetite, serum albumin), (viii) pain control or use of analgesics, and (ix) CRP / albumin ratio.

[0544] Treatment with the non-polymorphic form, crystalline form, or crystalline salt form of compound 1 combined with a checkpoint inhibitor or PD-1 inhibitor generates a more complex immunity that includes immunomodulation, which more efficiently restores appropriate immune function as well as the development of innate immunity and type 1 immunity.

[0545] Combination of Compound 1 and a Checkpoint Inhibitor

[0546] Combination of Compound 1 and Atezolizumab

[0547] In the aforementioned aspects and embodiments, compound 1 is administered to treat cancer together with a checkpoint inhibitor and optionally an additional immunomodulator.

[0548] In one embodiment, the checkpoint inhibitor is atezolizumab.

[0549] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0550] (i) administering a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising compound 1 to a subject, and

[0551] (ii) A step of administering a therapeutically effective amount of atezolizumab to the subject.

[0552] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0553] (i) administering to a subject a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof or compound 1 in a dose of about 5 mg to about 100 mg and

[0554] (ii) A step of administering a therapeutically effective amount of atezolizumab to the subject.

[0555] In another embodiment, the present invention comprises a method for treating cancer in a subject, wherein the method comprises a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound (1) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient,

[0556] The method includes the step of administering a therapeutically effective amount of atezolizumab or a pharmaceutical composition containing atezolizumab in combination to a subject requiring such treatment.

[0557] In another embodiment, the present invention comprises a method for treating cancer in a subject, wherein the method comprises a compound 1 or a pharmaceutically acceptable salt thereof in a dose of about 5 mg to about 100 mg, or a pharmaceutical composition comprising compound (1) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient,

[0558] The method includes the step of administering a therapeutically effective amount of atezolizumab or a pharmaceutical composition containing atezolizumab in combination to a subject requiring such treatment.

[0559] In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1, and atezolizumab or a pharmaceutical composition comprising atezolizumab are administered simultaneously, sequentially, or individually.

[0560] In one embodiment, the cancer is selected from heart cancer, head and neck cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, non-clear cell renal cell carcinoma, advanced clear cell renal cell carcinoma, castration-resistant prostate cancer, hormone receptor-positive breast cancer, prostate cancer, colon cancer, gastrointestinal cancer, breast cancer, genitourinary cancer, liver cancer, bone cancer, thyroid cancer, nervous system cancer, gynecological cancer, hematological cancer, skin cancer, urothelial carcinoma, and adrenal cancer.

[0561] In one embodiment, the amount of compound 1 or a pharmaceutically acceptable salt thereof administered is greater than 0.0 mg and less than or equal to 100 mg of compound 1; greater than 0.0 mg and less than or equal to 95 mg of compound 1; greater than 0.0 mg and less than or equal to 90 mg of compound 1; greater than 0.0 mg and less than or equal to 85 mg of compound 1; greater than 0.0 mg and less than or equal to 80 mg of compound 1; greater than 0.0 mg and less than or equal to 75 mg of compound 1; greater than 0.0 mg and less than or equal to 70 mg of compound 1; greater than 0.0 mg and less than or equal to 65 mg of compound 1; greater than 0.0 mg and less than or equal to 60 mg of compound 1; greater than 0.0 mg and less than or equal to 55 mg of compound 1; greater than 0.0 mg and less than or equal to 50 mg of compound 1; greater than 0.0 mg and less than or equal to 45 mg of compound 1; greater than 0.0 mg and less than or equal to 40 mg of compound 1; greater than 0.0 mg and less than or equal to 35 mg of compound 1; More than 0.0 mg and less than or equal to 30 mg of compound 1; more than 0.0 mg and less than or equal to 25 mg of compound 1; more than 0.0 mg and less than or equal to 20 mg of compound 1; more than 0.0 mg and less than or equal to 15 mg of compound 1; more than 0.0 mg and less than or equal to 10 mg of compound 1; or 5 mg or less of compound 1. In one embodiment, compound 1 is administered once a day. In another embodiment, compound 1 is administered twice a day.

[0562] In one embodiment, atezolizumab is administered intravenously (IV) to the subject. In another embodiment, atezolizumab is administered to the subject by parenteral injection.

[0563] In one embodiment, atezolizumab is administered once every 2 weeks, once every 3 weeks, or once every 4 weeks during the treatment period. In an additional embodiment, atezolizumab is administered once every 2 weeks during the treatment period. In another additional embodiment, atezolizumab is administered once every 3 weeks during the treatment period. In yet another additional embodiment, atezolizumab is administered once every 4 weeks during the treatment period.

[0564] In one embodiment, the dosage of atezolizumab is about 800 mg to about 1700 mg.

[0565] In one embodiment, the dosage of atezolizumab is about 840 mg administered once every 2 weeks, about 1200 mg administered once every 3 weeks, or 1680 mg administered once every 4 weeks.

[0566] In one embodiment, atezolizumab is administered to a subject in the form of an IV unit dose, wherein the dose comprises 840 mg, 1200 mg, or 1680 mg of atezolizumab, water, glacial acetic acid, L-histidine, polysorbate 20, and sucrose.

[0567] In one embodiment, atezolizumab is administered to a subject in the form of an IV unit dose, and the dosage form is sold as Tecentriq®.

[0568] Combination of Compound 1 and Avelumab

[0569] In one embodiment, the checkpoint inhibitor is avelumab.

[0570] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0571] (i) administering a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising compound 1 to a subject, and

[0572] (ii) A step of administering a therapeutically effective amount of avelumab to the subject.

[0573] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0574] (i) administering to a subject a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof or compound 1 in a dose of about 5 mg to about 100 mg and

[0575] (ii) A step of administering a therapeutically effective amount of avelumab to the subject.

[0576] In another embodiment, the present invention comprises a method for treating cancer in a subject, wherein the method comprises a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

[0577] The method includes the step of administering a therapeutically effective amount of avelumab or a pharmaceutical composition containing avelumab in combination to a subject requiring such treatment.

[0578] In another embodiment, the present invention comprises a method for treating cancer in a subject, wherein the method comprises a pharmaceutical composition comprising a compound 1 or a pharmaceutically acceptable salt thereof in a dose of about 5 mg to about 100 mg, or a compound 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

[0579] The method includes the step of administering a therapeutically effective amount of avelumab or a pharmaceutical composition containing avelumab in combination to a subject requiring such treatment.

[0580] In one embodiment, the present invention comprises a method for treating urothelial carcinoma in a subject, the method comprising:

[0581] (i) administering to a subject a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof or compound 1 in a dose of about 5 mg to about 100 mg and

[0582] (ii) A step of administering a therapeutically effective amount of avelumab to the subject.

[0583] In another embodiment, the present invention comprises a method for treating urothelial carcinoma in a subject, wherein the method comprises a pharmaceutical composition comprising a compound 1 or a pharmaceutically acceptable salt thereof in a dose of about 5 mg to about 100 mg, or a compound 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

[0584] The method includes the step of administering a therapeutically effective amount of avelumab or a pharmaceutical composition containing avelumab in combination to a subject requiring such treatment.

[0585] In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1, and avelumab or a pharmaceutical composition comprising avelumab are administered simultaneously, sequentially, or individually.

[0586] In one embodiment, the cancer is selected from heart cancer, head and neck cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, non-clear cell renal cell carcinoma, advanced clear cell renal cell carcinoma, castration-resistant prostate cancer, hormone receptor-positive breast cancer, prostate cancer, colon cancer, gastrointestinal cancer, breast cancer, genitourinary cancer, liver cancer, bone cancer, thyroid cancer, nervous system cancer, gynecological cancer, hematological cancer, skin cancer, urothelial carcinoma, and adrenal cancer.

[0587] In one embodiment, the urothelial carcinoma is a local advanced or metastatic transitional carcinoma of the urothelium. In another embodiment, the urothelial carcinoma is an advanced urothelial carcinoma. In yet another embodiment, the urothelial carcinoma is a urothelial carcinoma of the renal pelvis, ureter, bladder, or urethra.

[0588] In one embodiment, the amount of compound 1 or a pharmaceutically acceptable salt thereof administered is greater than 0.0 mg and less than or equal to 100 mg of compound 1; greater than 0.0 mg and less than or equal to 95 mg of compound 1; greater than 0.0 mg and less than or equal to 90 mg of compound 1; greater than 0.0 mg and less than or equal to 85 mg of compound 1; greater than 0.0 mg and less than or equal to 80 mg of compound 1; greater than 0.0 mg and less than or equal to 75 mg of compound 1; greater than 0.0 mg and less than or equal to 70 mg of compound 1; greater than 0.0 mg and less than or equal to 65 mg of compound 1; greater than 0.0 mg and less than or equal to 60 mg of compound 1; greater than 0.0 mg and less than or equal to 55 mg of compound 1; greater than 0.0 mg and less than or equal to 50 mg of compound 1; greater than 0.0 mg and less than or equal to 45 mg of compound 1; greater than 0.0 mg and less than or equal to 40 mg of compound 1; greater than 0.0 mg and less than or equal to 35 mg of compound 1; More than 0.0 mg and less than or equal to 30 mg of compound 1; more than 0.0 mg and less than or equal to 25 mg of compound 1; more than 0.0 mg and less than or equal to 20 mg of compound 1; more than 0.0 mg and less than or equal to 15 mg of compound 1; more than 0.0 mg and less than or equal to 10 mg of compound 1; or 5 mg or less of compound 1. In one embodiment, compound 1 is administered once a day. In another embodiment, compound 1 is administered twice a day.

[0589] In one embodiment, avelumab is administered intravenously (IV) to a subject. In another embodiment, avelumab is administered to a subject by intravenous infusion.

[0590] In one embodiment, avelumab is administered once every 2 weeks, once every 3 weeks, or once every 4 weeks during the treatment period. In an additional embodiment, avelumab is administered once every 2 weeks during the treatment period. In another additional embodiment, avelumab is administered once every 3 weeks during the treatment period. In yet another additional embodiment, avelumab is administered once every 4 weeks during the treatment period.

[0591] In one embodiment, the dosage of avelumab is about 500 mg to about 1700 mg.

[0592] In one embodiment, the dosage of avelumab is about 800 mg administered once every 2 weeks, about 1200 mg administered once every 3 weeks, or 1600 mg administered once every 4 weeks.

[0593] In one embodiment, the dosage of avelumab is about 800 mg administered once every two weeks.

[0594] In one embodiment, avelumab is administered to a subject in the form of an IV unit dose, and the form is sold as Bavencio®.

[0595] In one embodiment, the subject is a human.

[0596] In one embodiment, a subject with advanced urothelial carcinoma has stage IV disease according to the AJCC TNM staging criteria (8th edition, January 1, 2018).

[0597] In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1, and avelumab or a pharmaceutical composition comprising avelumab are administered simultaneously, sequentially, or individually.

[0598] In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1, and a combination of avelumab or a pharmaceutical composition comprising avelumab are administered as maintenance therapy to a subject with advanced urothelial carcinoma.

[0599] In one embodiment, a subject with advanced urothelial carcinoma received first-line platinum-based dual chemotherapy before maintenance therapy.

[0600] In one embodiment, the primary platinum-based dual chemotherapy includes gemcitabine + cisplatin and / or gemcitabine + carboplatin.

[0601] In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof, or a combination of a pharmaceutical composition comprising compound 1 and avelumab or a pharmaceutical composition comprising avelumab is administered as a second-line or third-line therapy to a subject with advanced urothelial carcinoma.

[0602] In one embodiment, a subject with advanced urothelial carcinoma received first-line platinum-based dual chemotherapy before second or third-line therapy.

[0603] In one embodiment, the primary platinum-based dual chemotherapy includes gemcitabine + cisplatin and / or gemcitabine + carboplatin.

[0604] In one embodiment, a subject with advanced urothelial carcinoma received primary platinum-based dual chemotherapy for at least 4 cycles but no more than 6 cycles.

[0605] In one embodiment, the subject was advanced after primary platinum-based dual chemotherapy.

[0606] In one embodiment, the method comprises the treatment of cancer that has not previously been treated with neoadjuvant immunotherapy using IL-2, IFN-α, or any anti-PD-1, anti-PD-L1, anti-PD-L2, anti-CD137, or CTLA-4 antibody (including ipilimumab), or any other antibody or drug that specifically targets T-cell co-stimulation or immune checkpoints. In another embodiment, the method comprises the treatment of cancer that has not previously been treated with a PD-1 or PD-L1 inhibitor.

[0607] Combination of Compound 1 and Nivolumab

[0608] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0609] (i) administering a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising compound 1 to a subject, and

[0610] (ii) A step of administering a therapeutically effective amount of nivolumab to the subject.

[0611] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0612] (i) administering to a subject a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof or compound 1 in a dose of about 5 mg to about 100 mg and

[0613] (ii) A step of administering a therapeutically effective amount of nivolumab to the subject.

[0614] In another embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising the step of administering to a subject requiring such treatment a pharmaceutical composition comprising a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof or compound 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient, in combination with a therapeutically effective amount of nivolumab or a pharmaceutical composition comprising nivolumab.

[0615] In another embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising the step of administering to a subject requiring such treatment a pharmaceutical composition comprising a dose of about 5 mg to about 100 mg of compound 1 or a pharmaceutically acceptable salt thereof or compound 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient, in combination with a pharmaceutical composition comprising a therapeutically effective amount of nivolumab or nivolumab.

[0616] In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1, and nivolumab or a pharmaceutical composition comprising nivolumab are administered simultaneously, sequentially, or individually.

[0617] In these and other embodiments, nivolumab is administered at a dose of about 360 mg every 3 weeks.

[0618] In one embodiment, the cancer is selected from melanoma, heart cancer, head and neck cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, non-clear cell renal cell carcinoma, advanced clear cell renal carcinoma, castration-resistant prostate cancer, hormone receptor-positive breast cancer, prostate cancer, colon cancer, gastrointestinal cancer, breast cancer, genitourinary cancer, liver cancer, bone cancer, thyroid cancer, nervous system cancer, gynecological cancer, hematological cancer, skin cancer, and adrenal cancer. In another embodiment, the cancer is selected from melanoma, non-small cell lung cancer, small cell lung cancer, renal cell carcinoma, classic Hodgkin lymphoma, head and neck squamous cell carcinoma, urothelial carcinoma, colorectal cancer with high microsatellite instability, and hepatocellular carcinoma.

[0619] In one embodiment, the cancer is a solid tumor. In another embodiment, the solid tumor is selected from the group consisting of sarcoma, carcinoma, and lymphoma. In a further embodiment, the solid tumor is a genitourinary cancer. In a further embodiment, the genitourinary cancer is selected from the group consisting of clear cell renal cell carcinoma (ccRCC), non-clear cell renal cell carcinoma (nccRCC), urothelial carcinoma (UC, ICI inexperienced and experienced), and metastatic castration-resistant prostate cancer (mCRPC).

[0620] The amount of Compound 1 or its pharmaceutically acceptable salt administered is greater than 0.0 mg and less than or equal to 100 mg of Compound 1; greater than 0.0 mg and less than or equal to 95 mg of Compound 1; greater than 0.0 mg and less than or equal to 90 mg of Compound 1; greater than 0.0 mg and less than or equal to 85 mg of Compound 1; greater than 0.0 mg and less than or equal to 80 mg of Compound 1; greater than 0.0 mg and less than or equal to 75 mg of Compound 1; greater than 0.0 mg and less than or equal to 70 mg of Compound 1; greater than 0.0 mg and less than or equal to 65 mg of Compound 1; greater than 0.0 mg and less than or equal to 60 mg of Compound 1; greater than 0.0 mg and less than or equal to 55 mg of Compound 1; greater than 0.0 mg and less than or equal to 50 mg of Compound 1; greater than 0.0 mg and less than or equal to 45 mg of Compound 1; greater than 0.0 mg and less than or equal to 40 mg of Compound 1; greater than 0.0 mg and less than or equal to 35 mg of Compound 1; More than 0.0 mg and less than or equal to 30 mg of compound 1; more than 0.0 mg and less than or equal to 25 mg of compound 1; more than 0.0 mg and less than or equal to 20 mg of compound 1; more than 0.0 mg and less than or equal to 15 mg of compound 1; more than 0.0 mg and less than or equal to 10 mg of compound 1; or 5 mg or less of compound 1. In one embodiment, compound 1 is administered once a day. In another embodiment, compound 1 is administered twice a day.

[0621] In one embodiment, nivolumab is administered intravenously (IV) to a subject. In another embodiment, nivolumab is administered to a subject by intravenous infusion.

[0622] In one embodiment, nivolumab is administered once every 2 weeks, once every 3 weeks, or once every 4 weeks during the treatment period. In an additional embodiment, nivolumab is administered once every 2 weeks during the treatment period. In another additional embodiment, nivolumab is administered once every 3 weeks during the treatment period. In yet another additional embodiment, nivolumab is administered once every 4 weeks during the treatment period.

[0623] In one embodiment, the dosage of nivolumab is about 50 mg to about 500 mg.

[0624] In one embodiment, nivolumab is administered to a subject in the form of an IV unit dose, and the dosage form is sold as OPDIVO®.

[0625] Combination of Compound 1 and another checkpoint inhibitor

[0626] In one embodiment, the checkpoint inhibitor is pembrolizumab.

[0627] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0628] (i) administering to a subject a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof or compound 1 in a dose of about 5 mg to about 100 mg and

[0629] (ii) A step of administering a therapeutically effective amount of pembrolizumab to the subject.

[0630] In another embodiment, the present invention comprises a method for treating cancer in a subject, wherein the method comprises a pharmaceutical composition comprising a compound 1 or a pharmaceutically acceptable salt thereof in a dose of about 5 mg to about 100 mg, or a compound 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

[0631] The method includes the step of administering a pharmaceutical composition containing pembrolizumab in combination to a subject requiring such treatment.

[0632] In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1, and pembrolizumab or a pharmaceutical composition comprising pembrolizumab are administered simultaneously, sequentially, or individually.

[0633] In one embodiment, the cancer is selected from melanoma, heart cancer, head and neck cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, non-clear cell renal cell carcinoma, advanced clear cell renal cell carcinoma, castration-resistant prostate cancer, hormone receptor-positive breast cancer, prostate cancer, colon cancer, gastrointestinal cancer, breast cancer, genitourinary cancer, liver cancer, bone cancer, thyroid cancer, nervous system cancer, gynecological cancer, hematological cancer, skin cancer, and adrenal cancer. In another embodiment, the cancer is selected from melanoma, non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, classic Hodgkin lymphoma, primary mediastinal B-cell lymphoma, cancer with high microsatellite instability, gastric cancer, esophageal cancer, cervical cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, urothelial carcinoma, or endometrial cancer.

[0634] The amount of Compound 1 or its pharmaceutically acceptable salt administered is greater than 0.0 mg and less than or equal to 100 mg of Compound 1; greater than 0.0 mg and less than or equal to 95 mg of Compound 1; greater than 0.0 mg and less than or equal to 90 mg of Compound 1; greater than 0.0 mg and less than or equal to 85 mg of Compound 1; greater than 0.0 mg and less than or equal to 80 mg of Compound 1; greater than 0.0 mg and less than or equal to 75 mg of Compound 1; greater than 0.0 mg and less than or equal to 70 mg of Compound 1; greater than 0.0 mg and less than or equal to 65 mg of Compound 1; greater than 0.0 mg and less than or equal to 60 mg of Compound 1; greater than 0.0 mg and less than or equal to 55 mg of Compound 1; greater than 0.0 mg and less than or equal to 50 mg of Compound 1; greater than 0.0 mg and less than or equal to 45 mg of Compound 1; greater than 0.0 mg and less than or equal to 40 mg of Compound 1; greater than 0.0 mg and less than or equal to 35 mg of Compound 1; More than 0.0 mg and less than or equal to 30 mg of compound 1; more than 0.0 mg and less than or equal to 25 mg of compound 1; more than 0.0 mg and less than or equal to 20 mg of compound 1; more than 0.0 mg and less than or equal to 15 mg of compound 1; more than 0.0 mg and less than or equal to 10 mg of compound 1; or 5 mg or less of compound 1. In one embodiment, compound 1 is administered once a day. In another embodiment, compound 1 is administered twice a day.

[0635] In one embodiment, pembrolizumab is administered intravenously (IV) to a subject. In another embodiment, pembrolizumab is administered to a subject by intravenous infusion.

[0636] In one embodiment, pembrolizumab is administered once every 2 weeks, once every 3 weeks, or once every 4 weeks during the treatment period. In an additional embodiment, pembrolizumab is administered once every 2 weeks during the treatment period. In another additional embodiment, pembrolizumab is administered once every 3 weeks during the treatment period. In yet another additional embodiment, pembrolizumab is administered once every 4 weeks during the treatment period.

[0637] In one embodiment, the dosage of pembrolizumab is about 50 mg to about 250 mg.

[0638] In one embodiment, pembrolizumab is administered to a subject in the form of an IV unit dose, and the form of administration is sold as KETRUDA®.

[0639] In one embodiment, the checkpoint inhibitor is durvalumab.

[0640] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0641] (i) administering to a subject a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof or compound 1 in a dose of about 5 mg to about 100 mg and

[0642] (ii) A step of administering a therapeutically effective amount of durvalumab to the subject.

[0643] In another embodiment, the present invention comprises a method for treating cancer in a subject, wherein the method comprises a pharmaceutical composition comprising a compound 1 or a pharmaceutically acceptable salt thereof in a dose of about 5 mg to about 100 mg, or a compound 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

[0644] The method includes the step of administering a therapeutically effective amount of durvalumab or a pharmaceutical composition containing durvalumab in combination to a subject requiring such treatment.

[0645] In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1, and durvalumab or a pharmaceutical composition comprising durvalumab are administered simultaneously, sequentially, or individually.

[0646] In one embodiment, the cancer is selected from melanoma, heart cancer, head and neck cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, non-clear cell renal cell carcinoma, advanced clear cell renal cell carcinoma, castration-resistant prostate cancer, hormone receptor-positive breast cancer, prostate cancer, colon cancer, gastrointestinal cancer, breast cancer, genitourinary cancer, liver cancer, bone cancer, thyroid cancer, nervous system cancer, gynecological cancer, hematological cancer, skin cancer, urothelial carcinoma, and adrenal cancer. In another embodiment, the cancer is selected from the group consisting of urothelial carcinoma and non-small cell lung cancer.

[0647] The amount of Compound 1 or its pharmaceutically acceptable salt administered is greater than 0.0 mg and less than or equal to 100 mg of Compound 1; greater than 0.0 mg and less than or equal to 95 mg of Compound 1; greater than 0.0 mg and less than or equal to 90 mg of Compound 1; greater than 0.0 mg and less than or equal to 85 mg of Compound 1; greater than 0.0 mg and less than or equal to 80 mg of Compound 1; greater than 0.0 mg and less than or equal to 75 mg of Compound 1; greater than 0.0 mg and less than or equal to 70 mg of Compound 1; greater than 0.0 mg and less than or equal to 65 mg of Compound 1; greater than 0.0 mg and less than or equal to 60 mg of Compound 1; greater than 0.0 mg and less than or equal to 55 mg of Compound 1; greater than 0.0 mg and less than or equal to 50 mg of Compound 1; greater than 0.0 mg and less than or equal to 45 mg of Compound 1; greater than 0.0 mg and less than or equal to 40 mg of Compound 1; greater than 0.0 mg and less than or equal to 35 mg of Compound 1; More than 0.0 mg and less than or equal to 30 mg of compound 1; more than 0.0 mg and less than or equal to 25 mg of compound 1; more than 0.0 mg and less than or equal to 20 mg of compound 1; more than 0.0 mg and less than or equal to 15 mg of compound 1; more than 0.0 mg and less than or equal to 10 mg of compound 1; or 5 mg or less of compound 1. In one embodiment, compound 1 is administered once a day. In another embodiment, compound 1 is administered twice a day.

[0648] In one embodiment, durvalumab is administered intravenously (IV) to the subject. In another embodiment, durvalumab is administered to the subject by parenteral injection.

[0649] In one embodiment, durvalumab is administered once every 2 weeks, once every 3 weeks, or once every 4 weeks during the treatment period. In an additional embodiment, durvalumab is administered once every 2 weeks during the treatment period. In another additional embodiment, durvalumab is administered once every 3 weeks during the treatment period. In yet another additional embodiment, durvalumab is administered once every 4 weeks during the treatment period.

[0650] In one embodiment, the dosage of durvalumab is about 10 mg / kg every 2 weeks.

[0651] In one embodiment, durvalumab is administered to a subject in the form of an IV unit dose, and the form is sold as IMFINZI®.

[0652] In one embodiment, the checkpoint inhibitor is semiplimab.

[0653] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0654] (i) administering to a subject a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof or compound 1 in a dose of about 5 mg to about 100 mg and

[0655] (ii) A step of administering a therapeutically effective amount of semiplimab or a pharmaceutically acceptable salt or prodrug thereof to the subject.

[0656] In another embodiment, the present invention comprises a method for treating cancer in a subject, wherein the method comprises a pharmaceutical composition comprising a compound 1 or a pharmaceutically acceptable salt thereof in a dose of about 5 mg to about 100 mg, or a compound 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

[0657] The method includes the step of administering a therapeutically effective amount of semiplimab or a pharmaceutical composition containing semiplimab to a subject requiring such treatment.

[0658] In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1, and semiplimab or a pharmaceutical composition comprising semiplimab are administered simultaneously, sequentially, or individually.

[0659] In one embodiment, the cancer is selected from melanoma, heart cancer, head and neck cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, non-clear cell renal cell carcinoma, advanced clear cell renal cell carcinoma, castration-resistant prostate cancer, hormone receptor-positive breast cancer, prostate cancer, colon cancer, gastrointestinal cancer, breast cancer, genitourinary cancer, liver cancer, bone cancer, thyroid cancer, nervous system cancer, gynecological cancer, hematological cancer, skin cancer, urothelial carcinoma, and adrenal cancer. In another embodiment, the cancer is cutaneous squamous cell carcinoma.

[0660] The amount of Compound 1 or its pharmaceutically acceptable salt administered is greater than 0.0 mg and less than or equal to 100 mg of Compound 1; greater than 0.0 mg and less than or equal to 95 mg of Compound 1; greater than 0.0 mg and less than or equal to 90 mg of Compound 1; greater than 0.0 mg and less than or equal to 85 mg of Compound 1; greater than 0.0 mg and less than or equal to 80 mg of Compound 1; greater than 0.0 mg and less than or equal to 75 mg of Compound 1; greater than 0.0 mg and less than or equal to 70 mg of Compound 1; greater than 0.0 mg and less than or equal to 65 mg of Compound 1; greater than 0.0 mg and less than or equal to 60 mg of Compound 1; greater than 0.0 mg and less than or equal to 55 mg of Compound 1; greater than 0.0 mg and less than or equal to 50 mg of Compound 1; greater than 0.0 mg and less than or equal to 45 mg of Compound 1; greater than 0.0 mg and less than or equal to 40 mg of Compound 1; greater than 0.0 mg and less than or equal to 35 mg of Compound 1; More than 0.0 mg and less than or equal to 30 mg of compound 1; more than 0.0 mg and less than or equal to 25 mg of compound 1; more than 0.0 mg and less than or equal to 20 mg of compound 1; more than 0.0 mg and less than or equal to 15 mg of compound 1; more than 0.0 mg and less than or equal to 10 mg of compound 1; or 5 mg or less of compound 1. In one embodiment, compound 1 is administered once a day. In another embodiment, compound 1 is administered twice a day.

[0661] In one embodiment, semiplimab is administered intravenously (IV) to the subject. In another embodiment, semiplimab is administered parenterally to the subject.

[0662] In one embodiment, semiplimab is administered once every 2 weeks, once every 3 weeks, or once every 4 weeks during the treatment period. In an additional embodiment, semiplimab is administered once every 2 weeks during the treatment period. In another additional embodiment, semiplimab is administered once every 3 weeks during the treatment period. In yet another additional embodiment, semiplimab is administered once every 4 weeks during the treatment period.

[0663] In one embodiment, the dosage of semiplimab is about 350 mg / 7 ml every 3 weeks.

[0664] In one embodiment, semiplimab is administered to a subject in the form of an IV unit dose, and the dosage form is sold as LIBTAYO®.

[0665] A combination of Compound 1, a checkpoint inhibitor, and an additional immunomodulator

[0666] In the aforementioned aspects and embodiments, compound 1 is administered together with a checkpoint inhibitor and optionally an additional immunomodulator to treat cancer. In one embodiment, the checkpoint inhibitor is selected from the group consisting of PD-1 inhibitors, PD-L1 inhibitors, and CTLA-4 inhibitors. In these and other aspects and embodiments, the checkpoint inhibitor is selected from the group consisting of pembrolizumab, nivolumab, atezolizumab (TECENTRIQ®), durvalumab, avelumab (BAVENCIO®), cemiplimab, camrelizumab, syntilimab, tisraelizumab, toripalizumab, spartalizumab, dostalimab, KN035 (Jiangshu Alphamab Biopharmaceuticals Company), cocibelimab (formerly CK-301), CA-170 (Kuris, Inc.), BMS-986189 (Bristol Myers Squibb Company), and ipilimumab (Yervoy, Bristol Myers Squibb Company).

[0667] In one embodiment, the checkpoint inhibitor is nivolumab.

[0668] In one embodiment, an additional immunomodulator is ipilimumab.

[0669] In another embodiment, the additional immunomodulator is an IL-2 targeting agent. In a further embodiment, the IL-2 targeting agent is a CD122-preferential IL-2 pathway agonist, a PEG-IL-2Rαβ-biased agonist, an IL-2Rβ-biased agonist, an IL-2Rβγ c - Biased agonist, IL-2v / IL-2α fusion protein, anti-EDB mAb(L19) / IL-2v fused to L19 / TNFv, anti-GD2 mAb / IL-2v, anti-FAP mAb / IL-2v, anti-CEA mAb / IL-2v, anti-PD-1 mAb / IL-2v, vaccine of patient-derived tumor cells + HD-IL-2, adoptive cell therapy + IL-2 infusion, adoptive cell therapy + IL-2 infusion + anti-PD-1 mAb, orthogonal IL-2v / IL-2Rβ mutant pair, anti-IL-2Rα mAb / PBD conjugate, PEG-IL-2Rα-biased agonist, IL-2v / human Fc fusion protein, PEG-IL-2Rα-biased(N88D) / IgG1 fusion protein, anti-IL-2 mAb / IL-2v, IL-2, It is selected from the group consisting of PPIs, recombinant plasmids encoding TGF-β1 and IL-10, and IL-2Rβ antagonists.

[0670] In one embodiment, the IL-2 targeting agent is a CD122-preferential IL-2 pathway agonist.

[0671] In one embodiment, the IL-2 targeting agent is bempegaldesleukin (BEMPEG; NKTR-214; Bristol Myers Squibb Company).

[0672] In one embodiment, IL - 2. The targeting agent is a PEG-IL-2Rαβ-biased agonist.

[0673] In one embodiment, the IL-2 targeting agent is THOR-707 (Sanofi).

[0674] In one embodiment, the IL-2 targeting agent is TransCon IL-2 β / γ (Ascendis Pharma).

[0675] In one embodiment, the IL-2 targeting agent is an IL-2Rβ-biased agonist.

[0676] In one embodiment, the IL-2 targeting agent is MDNA-19 (Medicenna).

[0677] In one embodiment, the IL-2 targeting agent is IL-2Rβγ c - It is a biased agonist.

[0678] In one embodiment, the IL-2 targeting agent is Neo-2 / 15 (Neoleukin).

[0679] In one embodiment, the IL-2 targeting agent is an IL-2v / IL-2Rα fusion protein.

[0680] In one embodiment, the IL-2 targeting agent is an anti-EDB mAb (L19) / IL-2v fused to L19 / TNFv.

[0681] In one embodiment, the IL-2 targeting agent is daromum (Philogen).

[0682] In one embodiment, the IL-2 targeting agent is anti-EDB mAb(L19) / IL-2v.

[0683] In one embodiment, the IL-2 targeting agent is darleukin (philogen).

[0684] In one embodiment, the IL-2 targeting agent is anti-GD2 mAb / IL-2v.

[0685] In one embodiment, the IL-2 targeting agent is APN-301 (APerion).

[0686] In one embodiment, the IL-2 targeting agent is RG-7461 (Roche).

[0687] In one embodiment, the IL-2 targeting agent is anti-CEA mAb / IL-2v.

[0688] In one embodiment, the IL-2 targeting agent is segutuzumab amanalukin (Roche).

[0689] In one embodiment, the IL-2 targeting agent is anti-PD-1 mAb / IL-2v.

[0690] In one embodiment, the IL-2 targeting agent is PD1-IL2v (Roche).

[0691] In one embodiment, the IL-2 targeting agent is a vaccine of patient-derived tumor cells + HD-IL-2.

[0692] In one embodiment, the IL-2 targeting agent is Oncoquest-L-Vaccine (Xemebiopharma.com).

[0693] In one embodiment, the IL-2 targeting agent is adoptive cell therapy + IL-2 injection.

[0694] In one embodiment, the IL-2 targeting agent is refillucel (Iovance).

[0695] In one embodiment, the IL-2 targeting agent is adoptive cell therapy + IL-2 infusion + anti-PD-1 mAb.

[0696] In one embodiment, the IL-2 targeting agent is refillucel + pembrolizumab.

[0697] In one embodiment, the IL-2 targeting agent is an orthogonal IL-2v / IL-2Rβ mutant pair.

[0698] In one embodiment, the IL-2 targeting agent is an anti-IL-2Rα mAb.PBD conjugate.

[0699] In one embodiment, the IL-2 targeting agent is kamidanlumab tesirin (ADC therapeutic).

[0700] In one embodiment, the IL-2 targeting agent is a PEG-IL2-Rα-biased agonist.

[0701] In one embodiment, the IL-2 targeting agent is NKTR-358 (Bristol Myers Squibb).

[0702] In one embodiment, the IL-2 targeting agent is THOR-809 (Sanofi).

[0703] In one embodiment, the IL-2 targeting agent is an IL-2v / human fusion protein.

[0704] In one embodiment, the IL-2 targeting agent is efavaleukin alpha (AMG592) (Amgen).

[0705] In one embodiment, the IL-2 targeting agent is an IL-2Rα-biased (N88D) / IgG1 fusion protein.

[0706] In one embodiment, the IL-2 targeting agent is RG-7835 (RO7049665) (Roche).

[0707] In one embodiment, the IL-2 targeting agent is an IL-2 mutain / Fc fusion protein.

[0708] In one embodiment, the IL-2 targeting agent is CC-92252 (Bristol Myers Squibb).

[0709] In one embodiment, the IL-2 targeting agent is anti-IL-2 mAb / IL-2v.

[0710] In one embodiment, the IL-2 targeting agent is F5111.2 (Creative Biolabs).

[0711] In one embodiment, the IL-2 targeting agent is a recombinant plasmid encoding IL-2, PPI, TGF-β1, and IL-10.

[0712] In one embodiment, the IL-2 targeting agent is NNC0361-0041 (NIDDK).

[0713] In one embodiment, the IL-2 targeting agent is an IL-2Rβ antagonist.

[0714] In one embodiment, the IL-2 targeting agent is MDNA-209 (Medicena).

[0715] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0716] (i) administering a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising compound 1 to a subject;

[0717] (ii) administering a therapeutically effective amount of nivolumab or a pharmaceutical composition comprising nivolumab to a subject; and

[0718] (iii) a step of administering a therapeutically effective amount of an immunomodulator or a pharmaceutical composition comprising a therapeutically effective amount of an immunomodulator to a subject.

[0719] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0720] (i) administering to a subject a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof or compound 1 in a dose of about 5 mg to about 100 mg;

[0721] (ii) administering a therapeutically effective amount of nivolumab or a pharmaceutical composition comprising nivolumab to a subject; and

[0722] (iii) a step of administering a therapeutically effective amount of an immunomodulator or a pharmaceutical composition comprising a therapeutically effective amount of an immunomodulator to a subject.

[0723] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0724] (i) administering a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising compound 1 to a subject;

[0725] (ii) administering a therapeutically effective amount of nivolumab or a pharmaceutical composition comprising nivolumab to a subject; and

[0726] (iii) a step of administering a therapeutically effective amount of ipilimumab or a pharmaceutical composition containing a therapeutically effective amount of ipilimumab to a subject.

[0727] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0728] (i) administering to a subject a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof or compound 1 in a dose of about 5 mg to about 100 mg;

[0729] (ii) administering a therapeutically effective amount of nivolumab or a pharmaceutical composition comprising nivolumab to a subject; and

[0730] (iii) a step of administering a therapeutically effective amount of ipilimumab or a pharmaceutical composition containing a therapeutically effective amount of ipilimumab to a subject.

[0731] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0732] (i) administering a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising compound 1 to a subject;

[0733] (ii) administering a therapeutically effective amount of nivolumab or a pharmaceutical composition comprising nivolumab to a subject; and

[0734] (iii) a step of administering a therapeutically effective amount of BEMPEG or a pharmaceutical composition containing a therapeutically effective amount of BEMPEG to a subject.

[0735] In one embodiment, the present invention comprises a method for treating cancer in a subject, the method comprising:

[0736] (i) administering to a subject a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof or compound 1 in a dose of about 5 mg to about 100 mg;

[0737] (ii) administering a therapeutically effective amount of nivolumab or a pharmaceutical composition comprising nivolumab to a subject; and

[0738] (iii) a step of administering a therapeutically effective amount of BEMPEG or a pharmaceutical composition containing a therapeutically effective amount of BEMPEG to a subject.

[0739] In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1, a pharmaceutical composition comprising nivolumab or nivolumab, and an pharmaceutical composition comprising ipilimumab or ipilimumab are administered simultaneously, sequentially, or individually.

[0740] In one embodiment, compound 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound 1, nivolumab or a pharmaceutical composition comprising nivolumab, and BEMPEG or a pharmaceutical composition comprising BEMPEG are administered simultaneously, sequentially, or individually.

[0741] In these and other embodiments, nivolumab is administered at about 360 mg IV every 3 weeks or about 240 mg IV every 2 weeks.

[0742] In these and other embodiments, ipilimumab is administered as a 3-time IV dose of about 1 mg / kg IV every 3 weeks.

[0743] In these and other embodiments, BEMPEG is administered at about 0.003 mg / kg IV every 2 weeks, about 0.006 mg / kg IV every 3 weeks, or about 0.009 mg / kg IV every 3 weeks.

[0744] In one embodiment, the cancer is selected from melanoma, heart cancer, head and neck cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, non-clear cell renal cell carcinoma, advanced clear cell renal carcinoma, castration-resistant prostate cancer, hormone receptor-positive breast cancer, prostate cancer, colon cancer, gastrointestinal cancer, breast cancer, genitourinary cancer, liver cancer, bone cancer, thyroid cancer, nervous system cancer, gynecological cancer, hematological cancer, skin cancer, and adrenal cancer. In another embodiment, the cancer is selected from melanoma, non-small cell lung cancer, small cell lung cancer, renal cell carcinoma, classic Hodgkin lymphoma, head and neck squamous cell carcinoma, urothelial carcinoma, colorectal cancer with high microsatellite instability, and hepatocellular carcinoma.

[0745] In one embodiment, the cancer is selected from heart cancer, head and neck cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, non-clear cell renal cell carcinoma, advanced or metastatic clear cell renal cell carcinoma, castration-resistant prostate cancer, hormone receptor-positive breast cancer, prostate cancer, colon cancer, gastrointestinal cancer, breast cancer, genitourinary cancer, liver cancer, bone cancer, thyroid cancer, nervous system cancer, gynecological cancer, hematological cancer, skin cancer, urothelial carcinoma, adrenal cancer, endometrial cancer, sarcoma, neuroendocrine tumor, ovarian cancer, hepatocellular carcinoma, gastric cancer, colorectal cancer, and melanoma.

[0746] In one embodiment, the cancer is selected from endometrial cancer, sarcoma, neuroendocrine tumor, ovarian cancer, colorectal cancer, HCC, NSCLC, gastric cancer, and melanoma.

[0747] In one embodiment, the cancer is a solid tumor. In another embodiment, the solid tumor is selected from the group consisting of sarcoma, carcinoma, and lymphoma. In a further embodiment, the solid tumor is a genitourinary cancer. In a further embodiment, the genitourinary cancer is selected from the group consisting of clear cell renal cell carcinoma (ccRCC), non-clear cell renal cell carcinoma (nccRCC), urothelial carcinoma (UC, ICI inexperienced and experienced), and metastatic castration-resistant prostate cancer (mCRPC).

[0748] The amount of Compound 1 or its pharmaceutically acceptable salt administered is greater than 0.0 mg and less than or equal to 100 mg of Compound 1; greater than 0.0 mg and less than or equal to 95 mg of Compound 1; greater than 0.0 mg and less than or equal to 90 mg of Compound 1; greater than 0.0 mg and less than or equal to 85 mg of Compound 1; greater than 0.0 mg and less than or equal to 80 mg of Compound 1; greater than 0.0 mg and less than or equal to 75 mg of Compound 1; greater than 0.0 mg and less than or equal to 70 mg of Compound 1; greater than 0.0 mg and less than or equal to 65 mg of Compound 1; greater than 0.0 mg and less than or equal to 60 mg of Compound 1; greater than 0.0 mg and less than or equal to 55 mg of Compound 1; greater than 0.0 mg and less than or equal to 50 mg of Compound 1; greater than 0.0 mg and less than or equal to 45 mg of Compound 1; greater than 0.0 mg and less than or equal to 40 mg of Compound 1; greater than 0.0 mg and less than or equal to 35 mg of Compound 1; More than 0.0 mg and less than or equal to 30 mg of compound 1; more than 0.0 mg and less than or equal to 25 mg of compound 1; more than 0.0 mg and less than or equal to 20 mg of compound 1; more than 0.0 mg and less than or equal to 15 mg of compound 1; more than 0.0 mg and less than or equal to 10 mg of compound 1; or 5 mg or less of compound 1. In one embodiment, compound 1 is administered once a day. In another embodiment, compound 1 is administered twice a day.

[0749] In one embodiment, nivolumab is administered intravenously (IV) to a subject. In another embodiment, nivolumab is administered to a subject by intravenous infusion.

[0750] In one embodiment, nivolumab is administered once every 2 weeks, once every 3 weeks, or once every 4 weeks during the treatment period. In an additional embodiment, nivolumab is administered once every 2 weeks during the treatment period. In another additional embodiment, nivolumab is administered once every 3 weeks during the treatment period. In yet another additional embodiment, nivolumab is administered once every 4 weeks during the treatment period.

[0751] In one embodiment, the dosage of nivolumab is about 50 mg to about 500 mg.

[0752] In one embodiment, nivolumab is administered to a subject in the form of an IV unit dose, and the dosage form is sold as OPDIVO®.

[0753] In these and other embodiments, nivolumab is administered at about 3 mg / kg IV every 2 weeks.

[0754] In these and other embodiments, ipilimumab is administered as a 4-time IV dose of about 3 mg / kg IV every 3 weeks.

[0755] In some embodiments, nivolumab is administered at about 1 mg / kg IV every 3 weeks, and ipilimumab is administered at about 3 mg / kg IV on the same day for up to 4 doses.

[0756] In some embodiments, nivolumab is administered at about 3 mg / kg IV every 3 weeks, and ipilimumab is administered at about 1 mg / kg IV on the same day for 4 doses.

[0757] In some embodiments, nivolumab is administered at about 3 mg / kg IV every 3 weeks for 4 doses, followed by 480 mg every 4 weeks, and ipilimumab is administered at about 1 mg / kg IV every 3 weeks for 4 doses.

[0758] In some embodiments, nivolumab is administered at about 3 mg / kg IV every 2 weeks, and ipilimumab is administered at about 1 mg / kg IV every 6 weeks.

[0759] In some embodiments, nivolumab is administered at a dose of about 360 mg every 3 weeks, and ipilimumab is administered at a dose of about 1 mg / kg IV on the same day for 4 doses.

[0760] In some embodiments, nivolumab is administered at a dose of about 240 mg every 2 weeks, and BEMPEG is administered at a dose of about 0.006 mg / kg every 3 weeks.

[0761] In some embodiments, nivolumab is administered at a dose of about 240 mg every 2 weeks, and BEMPEG is administered at a dose of about 0.003 mg / kg every 2 weeks.

[0762] In some embodiments, nivolumab is administered at a dose of about 240 mg every 2 weeks, and BEMPEG is administered at a dose of about 0.006 mg / kg every 2 weeks.

[0763] In some embodiments, nivolumab is administered at a dose of about 360 mg every 3 weeks, and BEMPEG is administered at a dose of about 0.006 mg / kg every 3 weeks.

[0764] In some embodiments, nivolumab is administered at a dose of about 360 mg every 3 weeks, and BEMPEG is administered at a dose of about 0.009 mg / kg every 3 weeks.

[0765] Additional combination of Compound 1 and other immunomodulators

[0766] The non-polymorphic form, crystalline form, or crystalline salt form of Compound 1 as disclosed herein is administered concurrently with a checkpoint inhibitor or PD-1 inhibitor. This is hereinafter referred to as the “combination.” The combination may be administered with one or more additional therapies for the treatment of a disease or disorder associated with hyperplasia, e.g., cancer. One or more additional therapies include: (i) surgery; (ii) radiotherapy (e.g., gamma radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy, and whole-body radioisotopes); (iii) endocrine therapy; (iv) adjuvant therapy, immunotherapy, CAR T-cell therapy; and (v) other chemotherapy agents.

[0767] The term "concurrently administered" ("concurrently administering") refers to the simultaneous administration or separate sequential administration of any additional active pharmaceutical component or component, including combinations and cytotoxic agents and radiation therapy as disclosed herein. If administration is not simultaneous, the compounds are administered at close intervals. Furthermore, it is not important whether the compounds are administered in the same dosage form; for example, one compound may be administered topically and another compound may be administered orally.

[0768] Typically, any agent active against the disease or condition being treated may be co-administered. Examples of such agents for cancer treatment include, for instance, https: / / www.cancer.gov / about-cancer / treatment / drugs( Last visited on January 22, 2019) and literature [Cancer Principles and Practice of Oncology by VT Devita and S. Hellman (editors), 11 th It can be found in publicly available sources such as the edition (2018), Lippincott Williams & Wilkins Publishers. A person skilled in the art will be able to discern which combination of agents would be useful based on the specific characteristics of the relevant drugs and diseases.

[0769] In one embodiment, the treatment method comprises the simultaneous administration of one or more additional therapies, including a combination and immunotherapy. Immunotherapy (also referred to as biological response modifier therapy, biological agent therapy, biotherapy, immunotherapy, or biological therapy) is a treatment that uses a part of the immune system to fight a disease. Immunotherapy can help recognize cancer cells or enhance the response to cancer cells. Immunotherapy includes active and passive immunotherapy. Active immunotherapy stimulates the body's own immune system, whereas passive immunotherapy generally uses immune system components generated outside the body.

[0770] Examples of active immunotherapy include, but are not limited to, vaccines containing cancer vaccines, tumor cell vaccines (autologous or allogeneic), dendritic cell vaccines, antigen vaccines, anti-ideotype vaccines, DNA vaccines, viral vaccines, or interleukin-2 (IL-2) containing tumor-infiltrating lymphocyte (TIL) vaccines or lymphokine-activated killer (LAK) cell therapy.

[0771] Examples of passive immunotherapy include, but are not limited to, targeted therapies involving monoclonal antibodies and toxins. Monoclonal antibodies include naked antibodies and conjugated monoclonal antibodies (also referred to as tagged, labeled, or loaded antibodies). Naked monoclonal antibodies are not to which drugs or radioactive materials are attached, whereas conjugated monoclonal antibodies are linked, for example, to chemotherapy drugs (chemolabeled), radioactive particles (radiolabeled), or toxins (immunotoxins). Examples of such naked monoclonal antibody drugs include, for example, rituximab (Rituxan), an antibody against the CD20 antigen used to treat B-cell non-Hodgkin lymphoma; for example, trastuzumab (Herceptin), an antibody against the HER2 protein used to treat advanced breast cancer; for example, alemtuzumab (Campat), an antibody against the CD52 antigen used to treat B-cell chronic lymphocytic leukemia (B-CLL). Examples include, but are not limited to, cetuximab (Erbitux), an antibody against the EGFR protein used, for example, in combination with irinotecan to treat advanced colorectal and head and neck cancers; and bevacizumab (Avastin), an anti-angiogenic therapy acting against the VEGF protein used, for example, in combination with chemotherapy to treat metastatic colorectal cancer. Examples of conjugated monoclonal antibodies include the radiolabeled antibody ibritumomab Tusetan (Zevalin), which delivers radioactivity directly to cancerous B lymphocytes and is used, for example, to treat B-cell non-Hodgkin lymphoma; and the radiolabeled antibody tocitumomab (Bexxar), used, for example, to treat certain types of non-Hodgkin lymphoma. and includes, for example, the immunotoxin gemtuzumab ozogamicin (Mylotarg) used to treat acute myeloid leukemia (AML), but is not limited to these.BL22 is a radiolabeled antibody, for example, a conjugated monoclonal antibody for treating hair cell leukemia, for example, an immunotoxin for treating leukemia, lymphoma and brain tumors, for example, OncoScint for colorectal and ovarian cancer, and for example, ProstaScint for prostate cancer.

[0772] Additional examples of therapeutic antibodies that may be used include HERCEPTIN™ (trastuzumab), a humanized anti-HER2 monoclonal antibody for the treatment of patients with metastatic breast cancer (Genentech, California); REOPRO.RTM. (absiximab), a platelet anti-glycoprotein IIb / IIIa receptor for preventing thrombosis (Centocor); ZENAPAX™ (daclizumab), an immunosuppressive humanized anti-CD25 monoclonal antibody for the prevention of acute renal allograft rejection (Roche Pharmaceuticals, Switzerland); PANOREX™ (Glaxo Wellcome / Centocor), a murine anti-17-IA cell surface antigen IgG2a antibody; and BEC2 (ImClone System), a murine anti-ideotype (GD3epitope) IgG antibody. IMC-C225 (Imclone Systems), a chimeric anti-EGFR IgG antibody; VITAXIN™ (Applied Molecular Evolution / Medlmmune), a humanized anti-alpha V beta 3 integrin antibody; Campat 1H / ℓDP-03 (Leukosite), a humanized anti-CD52 IgG1 antibody; SMART M195 (Protein Design Lab / Kanebo), a humanized anti-CD33 IgG antibody; RITUXAN™ (IDEC Pharm / Genentech, Roche / Zettyaku), a chimeric anti-CD20 IgG1 antibody; LYMPHOCIDE™ (Immunomedics), a humanized anti-CD22 IgG antibody; LYMPHOCIDE™ Y-90 (Immunomedics); Lymphoscan (Tc-99m-labeled; Radiography; Immunomedics); Nuvion (against CD3; Protein Design Lab); CM3, a humanized anti-ICAM3 antibody (ICOS Pharm); IDEC-114, a primate-modified anti-CD80 antibody (ICOS Pharm / Mitsubishi);ZEVALIN™ (IDEC / Schering AG), a radioisotope-labeled murine anti-CD20 antibody; IDEC-131 (IDEC / Eisai), a humanized anti-CD40L antibody; IDEC-151 (IDEC), a primate-modified anti-CD4 antibody; IDEC-152 (IDEC / Seikagaku), a primate-modified anti-CD23 antibody; SMART anti-CD3 (Protein Design Lab), a humanized anti-CD3 IgG; 5G1.1 (Alexion Pharm), a humanized anti-complement factor 5 (C5) antibody; D2E7 (CAT / BASF), a humanized anti-TNF-alpha antibody; CDP870 (Celltech), a humanized anti-TNF-alpha Fab fragment; IDEC-151 (IDEC Pharm / SmithKline Beecham), a primate-modified anti-CD4 IgG1 antibody; MDX-CD4 (Medarex / Eisai / Genmab), a human anti-CD4 IgG antibody; CD20-streptavidin (+biotin-yttrium 90; NeoRx); In CDP571 (CellTech), a humanized anti-TNF-alpha IgG4 antibody; LDP-02 (Leucocyte / Genentech), a humanized anti-alpha4 beta7 antibody; Orthoclone OKT4A (Ortho Biotech), a humanized anti-CD4 IgG antibody; ANTOVA™ (Biogen), a humanized anti-CD40L IgG antibody; ANTEGREN™ (Elan), a humanized anti-VLA-4 IgG antibody; and the human anti-TGF-beta 2 antibody CAT-152 (Cambridge Ab Tech) is included, but not limited to, these. Others are provided in subsequent paragraphs.;

[0773] One or more additional therapies also include adjuvant immunotherapy. Examples are cytokines, e.g., granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte-colony stimulating factor (G-CSF), macrophage inflammatory protein (MIP)-1-alpha, interleukins (including IL-1, IL-2, IL-4, IL-6, IL-7, IL-12, IL-15, IL-18, IL-21, and IL-27), tumor necrosis factor (including TNF-alpha), and interferons (including IFN-alpha, IFN-beta, and IFN-gamma); aluminum hydroxide (alum); Bacille Calmette-Guerin (BCG); Keyhole limpet hemocyanin (KLH); incomplete Freund's adjuvant (IFA); QS-21; DETOX; levamisole; and dinitrophenyl (DNP) and combinations thereof, e.g., combinations of interleukins, e.g., other cytokines such as IFN-alpha and IL-2.

[0774] In various embodiments, one or more additional therapies may include one or more of the following: adoptive cell delivery, angiogenesis inhibitors, Calmette-Guerin therapy, biochemotherapy, cancer vaccines, chimeric antigen receptor (CAR) T-cell therapy, cytokine therapy, gene therapy, immune checkpoint modulators, immunoconjugates, radioconjugates, oncolytic virus therapy, or targeted drug therapy. Immunological therapies or immunotherapies are collectively referred to herein as “immunotherapies.”

[0775] The present disclosure provides a method for preventing, treating, reducing, inhibiting, or controlling a neoplasm, tumor, or cancer in a subject requiring such a method, comprising the step of administering a therapeutically effective amount of a combination and one or more additional therapies. In various embodiments, treatment with the combination and one or more additional therapies provides a cooperative effect, an additive effect, or a synergistic effect in reducing the number of cancer cells when treated in combination compared to each of the treatments alone. In some embodiments, treatment with the combination and one or more additional therapies results in synergistic anti-tumor activity and / or anti-tumor activity that is more potent than the additive effect of administering the non-polymorphic form, crystalline form, or crystalline salt form of compound 1 or the immunotherapeutic agent alone.

[0776] Human cancers possess numerous genetic and epigenetic alterations that generate neoantigens potentially recognizable by the immune system (Sjoblom et al. (2006) Science 314:268-74). The adaptive immune system, composed of T lymphocytes and B lymphocytes, possesses potent anticancer potential due to its broad ability to respond to various tumor antigens and its sophisticated specificity. Furthermore, the immune system exhibits significant plasticity and memory elements. Successfully utilizing all these characteristics of the adaptive immune system could make immunotherapy unique among all modes of cancer treatment.

[0777] In various embodiments, one or more additional therapies include: adoptive cell delivery, angiogenesis inhibitors, Calmette-Guerin therapy, biochemotherapy, cancer vaccines, chimeric antigen receptor (CAR) T-cell therapy, cytokine therapy, gene therapy, immune checkpoint modulators, e.g., immune checkpoint inhibitors, immunoconjugates, radioconjugates, oncolytic virus therapy, or targeted drug therapy.

[0778] In each of the specific embodiments of the aforementioned aspects, as well as in other aspects and embodiments described elsewhere in this specification, one or more additional therapies enhance the activity of the combination.

[0779] In the specific embodiments of each of the aforementioned aspects, as well as in other aspects and embodiments described elsewhere in this specification, one or more additional therapies are modulators of immune cells (e.g., T-cells, dendritic cells, natural killer cells, etc.) selected from agonists or activators of co-stimulatory molecules, wherein the modulators are monoclonal antibodies, bispecific antibodies comprising one or more immune checkpoint antigen-binding moieties, trispecific antibodies, or immune cell-binding multivalent antibodies / fusion proteins / composites known in the art. In some embodiments, the immunotherapeutic agent may be an antibody that modulates the co-stimulatory molecule and binds to an antigen on the surface of an immune cell or cancer cell. In each of these different embodiments, the antibody modifier may be a monoclonal antibody, a polyclonal antibody, a bispecific antibody, a trispecific or multispecific type antibody, a fusion protein or a fragment thereof, e.g., a dibody, a single-strand (sc)-dyabody (scFv)2, a minibody, a minibody, a Barnase-barstar, scFv-Fc, sc(Fab)2, a trimeric antibody conjugate, a triabody antibody conjugate, a trimerbody antibody conjugate, a tribody antibody conjugate, a collabody antibody conjugate, a (scFv-TNFa)3 or F(ab)3 / DNL antibody conjugate.

[0780] In each of the specific embodiments of the aforementioned aspects, as well as in other aspects and embodiments described elsewhere in this specification, one or more additional therapies are immunotherapies that modulate the immune response, e.g., checkpoint inhibitors or checkpoint agonists. In some embodiments, one or more additional therapies are immunotherapies that enhance the anti-tumor immune response. In some embodiments, one or more additional therapies are immunotherapies that increase cell-mediated immunity. In some embodiments, one or more additional therapies are immunotherapies that increase T-cell activity. In some embodiments, one or more additional therapies are immunotherapies that increase cytolytic T-cell (CTL) activity.

[0781] In some embodiments, one or more additional therapies may comprise molecules, e.g., binders, e.g., antibodies or functional fragments thereof that modulate (activate or inhibit) checkpoint proteins upon co-administration with atezolizumab. The checkpoint inhibitor may be any molecule, agent, therapeutic agent, and / or method that inhibits immune checkpoints and / or promotes immune checkpoint inhibitors by, e.g., by promoting an intrinsic immune checkpoint inhibitor; by inhibiting transcription factors involved in the expression of immune checkpoints; and / or by acting in conjunction with some additional exogenous factors. For example, the checkpoint inhibitor may include a therapeutic agent that inhibits transcription factors involved in the expression of immune checkpoint genes or promotes the expression of transcription factors for tumor-suppressor genes, e.g., BACH2 (Luan et al., (2016). Transcription Factors and Checkpoint Inhibitor Expression with Age: Markers of Immunosenescence. Blood, 128(22), 5983). Furthermore, checkpoint inhibitors may inhibit the transcription of immune checkpoint genes; the modification and / or processing of immune checkpoint mRNAs; the translation of immune checkpoint proteins; and / or the activation of molecules involved in immune or immune checkpoint pathways, e.g., PD-1 transcription factors such as HIF-1, STAT3, NF-κB, and AP-1, or general oncogenic pathways, e.g., JAK / STAT, RAS / ERK, or PI3K / AKT / mTOR (Zerdes et al., Genetic, transcriptional and post-translational regulation of the programmed death protein ligand 1 in cancer: biology and clinical correlations, Oncogene volume 37, pages 4639-4661 (2018), the entire disclosure of which is incorporated herein by reference).

[0782] Checkpoint inhibitors may include therapeutic agents, molecules, agents, and / or methods that regulate immune checkpoints at the transcriptional level using, for example, co-inhibition of RNA-interference pathways and / or post-transcriptional gene silencing (PTGS) (e.g., microRNA, miRNA; silencing RNA, small-interfering RNA, or short-interfering RNA (siRNA). Transcriptional regulation of checkpoint molecules has been shown to include mir-16, which has been shown to target the 3'UTRs of checkpoint mRNAs CD80, CD274 (PD-L1), and CD40 (Leibowitz et al., Post-transcriptional regulation of immune checkpoint genes by mir-16 in melanoma, Annals of Oncology (2017) 28; v428-v448). Mir-33a has also been shown to be involved in regulating PD-1 expression in lung adenocarcinoma (Boldini et al., Role of microRNA-33a in regulating the expression of PD-1 in lung adenocarcinoma, Cancer Cell Int. 2017; 17: 105, the entirety of which is incorporated herein by reference).

[0783] T-cell-specific aptamer-siRNA chimeras have been proposed as a highly specific method for inhibiting molecules in immune checkpoint pathways (Hossain et al., The aptamer-siRNA conjugates: reprogramming T cells for cancer therapy, Ther. Deliv. 2015 Jan; 6(1): 1-4, the entirety of which is incorporated herein by reference).

[0784] Alternatively, members of immune checkpoint pathways can be inhibited by using therapeutic agents that affect related pathways, e.g., metabolism. For example, an oversupply of the glycolysis intermediate pyruvate in the mitochondria of CAD macrophages promoted PD-L1 expression through the induction of the bone morphogenetic protein 4 / phosphorylated SMAD1 / 5 / IFN regulatory factor 1 (BMP4 / p-SMAD1 / 5 / IRF1) signaling pathway. Therefore, administering treatments that modulate metabolic pathways can lead to subsequent modulation of the immunosuppressive PD-1 / PD-L1 checkpoint pathway (Watanabe et al., Pyruvate controls the checkpoint inhibitor PD-L1 and suppresses T cell immunity, J Clin Invest. 2017 Jun 30; 127(7): 2725-2738).

[0785] Checkpoint immunity can be regulated through oncolytic viruses that selectively replicate within tumor cells and induce an acute immune response in the tumor-microenvironment; that is, by acting as gene vectors that deliver specific agents (e.g., antibodies, miRNA, siRNA, etc.) to cancer cells, and by influencing tumor lysis and the secretion of cytokines and chemokines, thereby acting synergistically with immune checkpoint inhibition (Shi et al., Cancer Immunotherapy: A Focus on the Regulation of Immune Checkpoints, Int J Mol Sci. 2018 May; 19(5): 1389). Currently, clinical trials using the following viruses as checkpoint inhibitors are underway: poliovirus, measles virus, adenovirus, poxvirus, herpes simplex virus (HSV), coxsackievirus, reovirus, Newcastle disease virus (NDV), T-VEC (herpes virus encoded by GM-CSF (granulocyte-macrophage colony-stimulating factor)) and H101 (cit. [Shi et al.]).

[0786] Checkpoint inhibitors can act at the translational level of checkpoint immunity. Since the translation of mRNA into protein represents a key event in the regulation of gene expression, inhibiting immune checkpoint translation is a method by which immune checkpoint pathways can be inhibited.

[0787] Inhibition of immune checkpoint pathways can occur at any stage of the immune checkpoint translation process. For example, drugs, molecules, agents, therapeutics, and / or methods can inhibit the initiation process (thereby recruiting the 40S ribosomal subunit to the 5' end of the mRNA and scanning the 5'UTR of the mRNA to the 3' end). Inhibition can occur by targeting the base-pairing of the anticodon of the initiator methionyl-transfer RNA (tRNA) (Met-tRNAi) with the start codon, or by targeting the recruitment of the 60S subunit to initiate amino acid elongation and sequential addition in the translation of immune-checkpoint-specific genes. Alternatively, checkpoint inhibitors target the ternary complex (TC), namely eukaryotic initiation factor (eIF)2 (or one or more of its α, β, and γ subunits); GTP; And by preventing the formation of Met-tRNAi, it can inhibit the gateway at the translation level.

[0788] Checkpoint inhibition may occur through the destabilization of eIF2α by excluding phosphorylation via protein kinase R (PKR), PERK, GCN2, or HRI, or by excluding TC from associating with 40S ribosomes and / or other initiation factors, thereby preventing the formation of the preinitiation complex (PIC); and inhibiting the eIF4F complex and / or its cap-binding protein eIF4E, scaffolding protein eIF4G, or eIF4A helicase. Methods for discussing translational control in cancer are discussed in the literature [Truitt et al., New frontiers in translational control of the cancer genome, Nat Rev Cancer. 2016 Apr 26; 16(5): 288-304], the entire content of which is referenced herein by reference.

[0789] Checkpoint inhibitors may also include therapeutic agents, molecules, agents, and / or methods that modulate immune checkpoints at the cellular and / or protein level by inhibiting immune checkpoint receptors, for example. Inhibition of the checkpoint may occur through the use of antibodies, antibody fragments, antigen-binding fragments, small molecules, and / or other drugs, agents, therapeutic agents, and / or methods.

[0790] Immune checkpoints refer to inhibitory pathways of the immune system responsible for maintaining self-tolerance and regulating the intensity of immune responses to minimize peripheral tissue damage. However, tumor cells can also activate immune checkpoints to reduce the effectiveness of the immune response against tumor tissue ('blocking' the immune response). Unlike most anticancer drugs, checkpoint inhibitors do not directly target target tumor cells; instead, they target lymphocyte receptors or their ligands to enhance the endogenous antitumor activity of the immune system (Pardoll, 2012, Nature Reviews Cancer 12:252-264).

[0791] In some embodiments, one or more additional therapies are a modulator of PD-1 activity, a modulator of PD-L1 activity, a modulator of PD-L2 activity, a modulator of CTLA-4 activity, a modulator of CD28 activity, a modulator of CD80 activity, a modulator of CD86 activity, a modulator of 4-1BB activity, a modulator of OX40 activity, a modulator of KIR activity, a modulator of Tim-3 activity, a modulator of LAG3 activity, a modulator of CD27 activity, a modulator of CD40 activity, a modulator of GITR activity, a modulator of TIGIT activity, a modulator of CD20 activity, a modulator of CD96 activity, a modulator of IDO1 activity, a cytokine, a chemokine, an interferon, an interleukin, a lymphokine, a member of the tumor necrosis factor (TNF) family, or an immunostimulating oligonucleotide. In some embodiments, the immune checkpoint modulator is, i.e., an inhibitor or antagonist or an activator or agonist, and is, for example, a CD28 modulator, a 4-1BB modulator, an OX40 modulator, a CD27 modulator, a CD80 modulator, a CD86 modulator, a CD40 modulator or a GITR modulator, a Lag-3 modulator, a 41BB modulator, a LIGHT modulator, a CD40 modulator, a GITR modulator, a TGF-beta modulator, a TIM-3 modulator, a SIRP-alpha modulator, a TIGIT modulator, a VSIG8 modulator, a BTLA modulator, a SIGLEC7 modulator, a SIGLEC9 modulator, an ICOS modulator, a B7H3 modulator, a B7H4 modulator, a FAS modulator and / or a BTNL2 modulator. In some embodiments, the immunotherapeutic agent is an immune checkpoint modulator as described above (e.g., a monoclonal antibody, a bispecific antibody comprising one or more immune checkpoint antigen binding moiety, a trispecific antibody, or an immune checkpoint modulator antibody in the form of an immune cell-binding multivalent antibody / fusion protein / composition known in the art).

[0792] In some embodiments, one or more additional therapies are agents that inhibit the activity of PD-1. In some embodiments, one or more additional therapies are agents that inhibit the activity of PD-L1 and / or PD-L2. In some embodiments, one or more additional therapies are agents that inhibit the activity of CTLA-4. In some embodiments, one or more additional therapies are agents that inhibit the activity of CD80 and / or CD86. In some embodiments, one or more additional therapies are agents that inhibit the activity of TIGIT. In some embodiments, one or more additional therapies are agents that inhibit the activity of KIR. In some embodiments, one or more additional therapies are agents that enhance or stimulate the activity of activating immune checkpoint receptors.

[0793] PD-1 (also known as PDCD1, CD279, and PDCD1) is a cell surface receptor that plays an important role in regulating the balance between stimulating and inhibitory signals in the immune system and maintaining peripheral tolerance (Ishida, Y et al. 1992 EMBO J. 11 3887; Kier, Mary E et al. 2008 Annu Rev Immunol 26 677-704; Okazaki, Taku et al. 2007 International Immunology 19 813-824). PD-1 is an inhibitory member of the immunoglobulin superfamily that is homologous to CD28. PD-1 is a monomeric type 1 transmembrane protein consisting of a single immunoglobulin variable-like extracellular domain and a cytoplasmic domain containing an immunoreceptor tyrosine-based inhibitory motif (ITIM) and an immunoreceptor tyrosine-based switch motif (ITSM). PD-1 expression can be induced in T cells, B cells, natural killer (NK) cells, and monocytes, for example, upon lymphocyte activation via T cell receptor (TCR) or B cell receptor (BCR) signaling (Kier, Mary E et al. 2008 Annu Rev Immunol 26 677-704; Agata, Y et al 1996 Int Immunol 8 765-72). PD-1 is a receptor for ligands CD80, CD86, PD-L1 (B7-H1, CD274), and PD-L2 (B7-DC, CD273), which are cell surface-expressed members of the B7 family (Freeman, Gordon et al. 2000 J Exp Med 192 1027; Latchman, Y et al. 2001 Nat Immunol 2: 261).Upon ligand binding, PD-1 recruits phosphatases, such as SHP-1 and SHP-2, to intracellular tyrosine motifs, subsequently dephosphorylating effector molecules activated by TCR or BCR signaling (Chemnitz, J et al. 2004 J Immunol 173: 945-954; Riley, James L 2009 Immunological Reviews 229: 114-125). In this way, PD-1 transmits inhibitory signals to T cells and B cells only when co-involved with TCR or BCR.

[0794] PD-1 has been shown to downregulate effector T cell responses through both endogenous and exogenous functional mechanisms. Inhibitory signaling via PD-1 induces a non-response state in T cells, preventing them from clonal expansion or the production of optimal levels of effector cytokines. PD-1 can also induce apoptosis in T cells through its ability to inhibit survival signals from co-stimulation, which reduces the expression of key anti-apoptotic molecules such as Bcl-XL (Kier, Mary E et al. 2008 Annu Rev Immunol 26: 677-704). In addition to these direct effects, recent publications have suggested that PD-1 is involved in the suppression of effector cells by promoting the induction and maintenance of regulatory T cells (TREGs). For example, PD-L1 expressed in dendritic cells was shown to act synergistically with TGF-β to promote the induction of CD4+ FoxP3+TREG with enhanced inhibitory function (Francisco, Loise M et al. 2009 J Exp Med 206: 3015-3029).

[0795] TIM-3 (also known as T-cell immunoglobulin and mucin-domain-containing-3, TIM-3, hepatitis A virus cell receptor 2, HAVCR2, HAVcr-2, KIM-3, TIMD-3, TIMD3, Tim-3, and CD366) is a single-pass type I membrane protein of approximately 33.4 kDa involved in immune responses (Sanchez-Fueyo et al., Tim-3 inhibits T helper type 1-mediated auto- and alloimmune responses and promotes immunological tolerance, Nat. Immunol. 4: 1093-1101(2003)).

[0796] TIM-3 is selectively expressed in Th1 cells and phagocytes (e.g., macrophages and dendritic cells). The use of siRNA or blocking antibodies to reduce human TIM-3 expression increased the secretion of interferon γ (IFN-γ) from CD4-positive T cells, suggesting an inhibitory role of TIM-3 in human T cells. Analysis of clinical samples from patients with autoimmune diseases showed that TIM-3 is not expressed in CD4-positive cells. In particular, TIM-3 expression levels were lower and IFN-γ secretion was higher in T cell clones derived from the cerebrospinal fluid of multiple sclerosis patients compared to clones derived from healthy individuals (Koguchi K et al., J Exp Med. 203: 1413-8. (2006)).

[0797] TIM-3 is a member of the galectin family, a molecule ubiquitously expressed in various cell types, and contains β-galactoside; phosphatidylserine (PtdSer) (DeKryff et al., T cell / transmembrane, Ig, and mucin-3 allelic variants differentially recognize phosphatidylserine and mediate phagocytosis of apoptotic cells, J Immunol. 2010 Feb 15; 184(4): 1918-30); high motility group protein 1 (also known as HMGB1, HMG1, HMG3, SBP-1, HMG-1, and high motility group box 1) (Chiba et al., Tumor-infiltrating DCs suppress nucleic acid-mediated innate immune responses through interactions between the receptor TIM-3 and the alarmin HMGB1, Nat Immunol. 2012 Sep; 13(9): 832-42); It is a receptor for the ligand galectin-9 that binds to carcinoma embryonic antigen-associated cell adhesion molecule 1 (also known as CEACAM1, BGP, BGP1, BGPI, and carcinoma embryonic antigen-associated cell adhesion molecule 1) (Huang et al., CEACAM1 regulates TIM-3-mediated tolerance and exhaustion, Nature. 2015 Jan 15; 517(7534): 386-90).

[0798] BTLA (also known as B-lymphocyte and T-lymphocyte attenuation factor, BTLA1, CD272, and B and T lymphocyte association) is a single-pass type I membrane protein of approximately 27.3-kDa involved in lymphocyte inhibition during immune responses. BTLA is constitutively expressed in both B cells and T cells. BTLA interacts with HVEM (herpes virus entry mediator), a member of the tumor-necrosis factor receptor (TNFR) family (Gonzalez et al., Proc. Natl. Acad. Sci. USA, 2005, 102: 1116-21). The interaction between BTLA, which belongs to the CD28 family of the immunoglobulin superfamily, and HVEM, a co-stimulating tumor-necrosis factor (TNF) receptor (TNFR), is unique in that it defines crosstalk between these two receptor families. BTLA contains a membrane-proximal immune receptor tyrosine-based inhibitory motif (ITIM) and an immune receptor tyrosine-based conversion motif (ITSM). Disruption of the ITIM or ITSM abolished BTLA’s ability to recruit SHP1 or SHP2, suggesting that BTLA recruits SHP1 and SHP2 in a manner different from PD-1 and that both tyrosine motifs are necessary to block T cell activation. The BTLA cytoplasmic tail also contains a third conserved tyrosine-containing motif within a cytoplasmic domain that is sequenced similarly to the Grb-2 recruitment site (YXN). In addition, phosphorylated peptides containing this BTLA N-terminal tyrosine motif can interact with the p85 subunits of GRB2 and PI3K in vitro, but the functional effects of this interaction remain unexplored in vivo (Gavrieli et al., Bioochem. Biophysi Res Commun, 2003, 312, 1236-43). BTLA is a receptor for the ligands PTPN6 / SHP-1; PTPN11 / SHP-2; TNFRSF14 / HVEM; and B7H4.

[0799] VISTA (also known as the V-domain Ig repressor of cell-activating VSIR, B7-H5, B7H5, GI24, PP2135, SISP1, DD1alpha, VISTA, C10orf54, chromosome 10 open reading frame 54, PD-1H, and V-set immunomodulatory receptor) is a single-pass type I membrane protein of approximately 33.9-kDa involved in T-cell inhibitory responses, embryonic stem cell differentiation through inhibition of BMP4 signaling, and MMP14-mediated MMP2 activation (Yoon et al., Control of signaling-mediated clearance of apoptotic cells by the tumor suppressor p53, Science. 2015 Jul 31; 349(6247): 1261669). VISTA interacts with the ligand VSIG-3 (Wang et al., VSIG-3 as a ligand of VISTA inhibits human T-cell function, Immunology. 2019 Jan; 156(1): 74-85).

[0800] LAG-3 (also known as lymphocyte-activation gene 3, LAG3, CD223, and lymphocyte activation 3) is a single-pass type I membrane protein of approximately 57.4-kDa involved in lymphocyte activation that also binds to HLA class-II antigens. LAG-3 is a member of the immunoglobulin supergene family and is involved in activated T cells (Huard et al., 1994, Immunogenetics 39: 213), NK cells (Triebel et al., 1990, J. Exp. Med. 171: 1393-1405), and regulatory T cells (Huang et al., 2004, Immunity 21: 503-513; Camisaschi et al., 2010, J Immunol. 184: 6545-6551; Gagliani et al., 2013, Nat Med 19: It is expressed in (739-746) and plasmacytic dendritic cells (DC) (Workman et al., 2009, J Immunol 182: 1885-1891). LAG-3 is a membrane protein encoded by a gene located on chromosome 12 and is structurally and genetically related to CD4. Like CD4, LAG-3 can interact with MHC class II molecules on the cell surface (Baixeras et al., 1992, J. Exp. Med. 176: 327-337; Huard et al., 1996, Eur. J. Immunol. 26: 1180-1186). Direct binding of LAG-3 to MHC class II plays a role in downregulating antigen-dependent stimulation of CD4+ T lymphocytes (Huard et al., 1994, Eur. J. Immunol. 24: 3216-3221), LAG-3 blockade is also effective against tumor or auto-antigens (Gross et al., 2007, J Clin Invest. 117: 3383-3392) and viral models (Blackburn et al., 2009, Nat. Immunol.(10: 29-37) It has been suggested that it activates CD8+ lymphocytes in all cases. Additionally, the cytoplasmic domain of LAG-3 can interact with LAP (LAG-3-associated protein), a signaling molecule involved in the downregulation of the CD3 / TCR activation pathway (Iouzalen et al., 2001, Eur. J. Immunol. 31: 2885-2891). Furthermore, CD4+CD25+ regulatory T cells (Treg) have been shown to express LAG-3, which contributes to the inhibitory activity of Treg cells upon activation (Huang, C. et al., 2004, Immunity 21: 503-513). LAG-3 can also negatively regulate T cell homeostasis by Treg cells through both T cell-dependent and independent mechanisms (Workman, CJ and Vignali, DA, 2005, J. Immunol. 174: 688-695).

[0801] LAG-3 has been shown to interact with MHC class II molecules (Huard et al., CD4 / major histocompatibility complex class II interaction analyzed with CD4- and lymphocyte activation gene-3 (LAG-3)-Ig fusion proteins, Eur J Immunol. 1995 Sep; 25(9): 2718-21).

[0802] Additionally, several kinases are known as checkpoint inhibitors. For example, CHEK-1, CHEK-2, and A2aR.

[0803] CHEK-1 (also known as CHK 1 kinase, CHK1, and checkpoint kinase 1) is a serine / threonine-protein kinase of approximately 54.4 kDa involved in the activation of checkpoint-mediated cell cycle arrest and DNA repair in response to DNA damage and / or non-replicated DNA.

[0804] CHEK-2 (also known as CHK2 kinase, CDS1, CHK2, HuCds1, LFS2, PP1425, RAD53, hCds1, and checkpoint kinase 2) is a serine / threonine protein kinase of approximately 60.9 kDa involved in checkpoint-mediated cell cycle arrest, DNA-repair activation, and double-strand break-mediated apoptosis.

[0805] A2aR (also known as adenosine A2A receptor, ADORA2A, adenosine A2a receptor, A2aR, ADORA2, and RDC8) is a multi-pass membrane receptor of about 44.7-kDa for adenosine and other ligands.

[0806] In some embodiments, the exemplary immunotherapeutic agent may comprise one or more antibody modulators targeting PD-1, PD-L1, PD-L2, CEACAM (e.g., CEACAM-1, -3 and / or -5), CTLA-4, TIM-3, LAG-3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4, TGF-beta, OX40, 41BB, LIGHT, CD40, GITR, TGF-beta, TIM-3, SIRP-alpha, VSIG8, BTLA, SIGLEC7, SIGLEC9, ICOS, B7H3, B7H4, FAS and / or BTNL2 among others known in the art. In some embodiments, the immunotherapeutic agent is an agent that increases natural killer (NK) cell activity. In some embodiments, one or more additional therapies are agents that inhibit the suppression of the immune response. In some embodiments, one or more additional therapies are inhibitory cells or agents that inhibit inhibitory cell activity. In some embodiments, one or more additional therapies are agents or therapies that inhibit Treg activity. In some embodiments, one or more additional therapies are agents that inhibit the activity of inhibitory immune checkpoint receptors.

[0807] In some embodiments, one or more additional therapies comprise a T cell modulator selected from an agonist or activator of a co-stimulatory molecule in co-administration with a checkpoint inhibitor. In one embodiment, the agonist of the co-stimulatory molecule is selected from an agonist of GITR, OX40, SLAM (e.g., SLAMF7), HVEM, LIGHT, CD2, CD27, CD28, CDS, ICAM-1, LFA-1 (CD11a / CD18), ICOS (CD278), 4-1BB (CD137), CD30, CD40, BAFFR, CD7, NKG2C, NKp80, CD160, B7-H3, or CD83 ligand (e.g., an acting antibody or its antigen-binding fragment or soluble fusion). In another embodiment, the effector cell combination comprises a bispecific T cell engaging antibody (e.g., a bispecific antibody molecule that binds to CD3 and tumor antigens (e.g., particularly EGFR, PSCA, PSMA, EpCAM, HER2)).

[0808] In some embodiments, one or more additional therapies are a modulator of PD-1 activity, a modulator of PD-L1 activity, a modulator of PD-L2 activity, a modulator of CTLA-4 activity, a modulator of CD28 activity, a modulator of CD80 activity, a modulator of CD86 activity, a modulator of 4-1BB activity, a modulator of OX40 activity, a modulator of KIR activity, a modulator of Tim-3 activity, a modulator of LAG3 activity, a modulator of CD27 activity, a modulator of CD40 activity, a modulator of GITR activity, a modulator of TIGIT activity, a modulator of CD20 activity, a modulator of CD96 activity, a modulator of IDO1 activity, a modulator of SIRP-alpha activity, a modulator of TIGIT activity, a modulator of VSIG8 activity, a modulator of BTLA activity, a modulator of SIGLEC7 activity, a modulator of SIGLEC9 activity, a modulator of ICOS activity, a modulator of B7H3 activity, a modulator of B7H4 activity, and FAS It is a modulator of activity, a modulator of BTNL2 activity, a cytokine, a chemokine, an interferon, an interleukin, a lymphokine, a member of the tumor necrosis factor (TNF) family, or an immunomodulatory oligonucleotide.

[0809] In some embodiments, one or more additional therapies are immune checkpoint modulators (e.g., immune checkpoint inhibitors, e.g., inhibitors of PD-1 activity, modulators of PD-L1 activity, modulators of PD-L2 activity, modulators of CTLA-4, or CD40 agonists (e.g., anti-CD40 antibody molecules), (xi) OX40 agonists (e.g., anti-OX40 antibody molecules), or (xii) CD27 agonists (e.g., anti-CD27 antibody molecules)). In one embodiment, the immunotherapeutic agent is an inhibitor of PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, CEACAM (e.g., CEACAM-1, -3 and / or -5), VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and / or TGF beta, galectin 9, CD69, galectin-1, CD113, GPR56, CD48, GARP, PD1H, LAIR1, TIM-1 and TIM-4. In one embodiment, the inhibitor of the immune checkpoint molecule inhibits PD-1, PD-L1, LAG-3, TIM-3, CEACAM (e.g., CEACAM-1, -3 and / or -5), CTLA-4 or any combination thereof.

[0810] In one embodiment, the immunotherapeutic agent is an agonist of a protein that stimulates T cell activation, such as B7-1, B7-2, CD28, 4-1BB(CD137), 4-1BBL, ICOS, ICOS-L, OX40, OX40L, GITR, GITRL, CD70, CD27, CD40, DR3, and CD28H.

[0811] In some embodiments, one or more additional therapies are activators or agonists of co-stimulatory molecules. In one embodiment, the agonist of the co-stimulatory molecule is selected from agonists of CD2, CD28, CDS, ICAM-1, LFA-1 (CD11a / CD18), ICOS (CD278), 4-1BB (CD137), GITR, CD30, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160, B7-H3, or CD83 ligands (e.g., agonist antibodies or their antigen-binding fragments or soluble fusions).

[0812] Inhibition of the inhibitory molecule may be performed at the DNA, RNA, or protein level. In an embodiment, an inhibitory nucleic acid (e.g., dsRNA, siRNA, or shRNA) may be used to inhibit the expression of the inhibitory molecule. In another embodiment, the inhibitor of the inhibitory signal is a polypeptide, e.g., a soluble ligand (e.g., PD-1-Ig or CTLA-4 Ig), or an antibody or its antigen-binding fragment, e.g., a monoclonal antibody, a bispecific antibody comprising one or more immune checkpoint antigen-binding moieties, a trispecific antibody, or an immune cell-binding multivalent antibody / fusion protein / composition known in the art that binds to the inhibitory molecule; For example, an antibody or fragment thereof that binds to PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, CEACAM (e.g., CEACAM-1, -3 and / or -5), VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and / or TGF beta, galectin 9, CD69, galectin-1, CD113, GPR56, CD48, GARP, PD1H, LAIR1, TIM-1, TIM-4 or a combination thereof (also referred to herein as “antibody molecule”).

[0813] In some embodiments, one or more additional therapies are monoclonal antibodies or bispecific antibodies co-administered with the checkpoint inhibitor. For example, the monoclonal or bispecific antibodies may specifically bind to members of the c-Met pathway and / or immune checkpoint modulators (e.g., bispecific antibodies such as antibodies binding to PD-1, PD-L1, PD-L2, or CTLA-4, LAG-3, OX40, 41BB, LIGHT, CD40, GITR, TGF-beta, TIM-3, SIRP-alpha, TIGIT, VSIG8, BTLA, SIGLEC7, SIGLEC9, ICOS, B7H3, B7H4, FAS, BTNL2, or CD27 bind to both the hepatocyte growth factor receptor (HGFR) and the immune checkpoint modulators described herein). In a specific embodiment, the bispecific antibody specifically binds to the human HGFR protein and one of PD-1, PD-L1, and CTLA-4.

[0814] In some embodiments of the method described herein, one or more additional therapies are PD-1 antagonists, PD-L1 antagonists, PD-L2 antagonists, CTLA-4 antagonists, CD80 antagonists, CD86 antagonists, KIR antagonists, Tim-3 antagonists, LAG3 antagonists, TIGIT antagonists, CD20 antagonists, CD96 antagonists, or IDO1 antagonists.

[0815] In some embodiments, the PD-1 antagonist is an antibody that specifically binds to PD-1. In some embodiments, the antibody binding to PD-1 is pembrolizumab (KEYTRUDA®, MK-3475; Merck), fidilizumab (CT-011; Curetech Ltd.), nivolumab (OPDIVO®, BMS-936558, MDX-1106; Bristol Myer Squibb), MEDI0680 (AMP-514; AstraZeneca / MedImmune), REGN2810 (Regeneron Pharmaceuticals), BGB-A317 (BeiGene Ltd.), PDR-001 (Novartis), or STI-A1110 (Sorrento Therapeutics Therapeutics. In some embodiments, an antibody binding to PD-1, e.g., an antibody identified as APE2058, APE1922, APE1923, APE1924, APE 1950, or APE1963 (Anaptysbio), or an antibody comprising the CDR region of any of these antibodies, is described in PCT Publication WO 2014 / 179664. In other embodiments, the PD-1 antagonist is a fusion protein comprising the extracellular domain of PD-L1 or PD-L2, e.g., AMP-224 (AstraZeneca / MedImmune). In other embodiments, the PD-1 antagonist is a peptide inhibitor, e.g., AUNP-12 (Aurigene).

[0816] In some embodiments, the PD-L1 antagonist is an antibody that specifically binds to PD-L1. In some embodiments, the antibody that binds to PD-L1 is MEDI4736 (AstraZeneca / MedImmune), BMS-936559 (MDX-1105; Bristol Myers Squibb), avelumab (MSB0010718C; Merck KGaA), KD033 (Kadmon), or the antibody portion of KD033 or STI-A1014 (Sorrento Therapeutics). In some embodiments, antibodies that bind to PD-L1, e.g., Ab-14, Ab-16, Ab-30, Ab-31, Ab-42, Ab-50, Ab-52 or Ab-55 or antibodies comprising a CDR region of any of these antibodies are described in PCT Publication WO 2014 / 055897, the entirety of which is incorporated herein by reference.

[0817] In some embodiments, the CTLA-4 antagonist is an antibody that specifically binds to CTLA-4. In some embodiments, the antibody that binds to CTLA-4 is ipilimumab (YERVOY®; Bristol Myers Squibb) or tremelimumab (CP-675, 206; Pfizer). In some embodiments, the CTLA-4 antagonist is a CTLA-4 fusion protein or a soluble CTLA-4 receptor, e.g., KARR-102 (Kahr Medical Ltd.).

[0818] In some embodiments, the LAG3 antagonist is an antibody that specifically binds to LAG3. In some embodiments, the antibodies that bind to LAG3 are IMP701 (Prima BioMed), IMP731 (Prima BioMed / GlaxoSmithKline), BMS-986016 (Bristol Myers Squibb), LAG525 (Novartis), and GSK2831781 (GlaxoSmithKline). In some embodiments, the LAG3 antagonist comprises a soluble LAG3 receptor, e.g., IMP321 (Prima BioMed).

[0819] In some embodiments, the KIR antagonist is an antibody that specifically binds to KIR. In some embodiments, the antibody that binds to KIR is ririlumab (Bristol Myers Squibb / Innate Pharma).

[0820] In some embodiments, the immunotherapeutic agent is a cytokine, e.g., a chemokine, interferon, interleukin, lymphokine, or a member of the tumor necrosis factor family. In some embodiments, the cytokine is IL-2, IL15, or interferon-gamma.

[0821] In any of the above embodiments or in some embodiments of the embodiments described elsewhere in this specification, cancer is lung cancer (e.g., non-small cell lung cancer (NSCLC)), kidney cancer (e.g., renal urothelial carcinoma), bladder cancer (e.g., bladder urothelial (transitional cell) carcinoma), breast cancer, colorectal cancer (e.g., colon adenocarcinoma), ovarian cancer, pancreatic cancer, gastric carcinoma, esophageal cancer, mesothelioma, melanoma (e.g., cutaneous melanoma), head and neck cancer (e.g., head and neck squamous cell carcinoma (HNSCC)), thyroid cancer, sarcoma (e.g., soft tissue sarcoma, fibrosarcoma, myxosarcoma, liposarcoma, osteogenic sarcoma, osteosarcoma, chondrosarcoma, angiosarcoma, endothelial sarcoma, lymphangisarcoma, lymphangiendothelial sarcoma, leiomyosarcoma or rhabdomyosarcoma), prostate cancer, glioblastoma, cervical cancer, thymic carcinoma, leukemia (e.g. For example, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), chronic eosinophilic leukemia or chronic lymphoblastic leukemia (CLL)), lymphoma (e.g., Hodgkin lymphoma or non-Hodgkin lymphoma (NHL)), myeloma (e.g., multiple myeloma (MM)), mycosarcoma, Merkel cell carcinoma, hematological malignancies, cancers of blood tissues, B-cell carcinoma, bronchial cancer, gastric cancer, brain cancer or central nervous system cancer, peripheral nervous system cancer, uterine cancer or endometrial cancer, oral cancer or pharyngeal cancer, liver cancer, testicular cancer, biliary tract cancer, small intestine cancer or appendiceal cancer, salivary gland cancer, adrenal cancer, adrenocortical carcinoma, adenocarcinoma, inflammatory myofibroblastic tumor, gastrointestinal stromal tumor (GIST), colon cancer, myelodysplastic syndrome (MDS), myeloproliferative disorder (MPD), polycythemia vera, Chordoma, synovial tumor, Ewing tumor, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, medullary carcinoma, bronchial carcinoma, renal cell carcinoma, liver tumor, cholangiocarcinoma, choriocarcinoma, seminoma, embryonic carcinoma, Wilms tumor, bladder carcinoma, epithelial carcinoma, glioma, anaplastic astrocytoma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pineal tumor, hemangioma, auditory tumor, oligodendroma, meningioma,It is selected from the group consisting of neuroblastoma, retinoblastoma, follicular lymphoma, diffuse large B-cell lymphoma, mantle cell lymphoma, hepatocellular carcinoma, thyroid cancer, small cell carcinoma, essential thrombocytosis, myeloid metaplasia, eosinophilia syndrome, systemic mastocytosis, familial hypereosinophilia, neuroendocrine cancer, or carcinoid tumor.

[0822] In any of the above embodiments or in some embodiments of the embodiments described elsewhere in this specification, the cancer or tumor of the subject does not respond to immune checkpoint inhibition (e.g., any immune checkpoint inhibitor described herein, such as a PD-1 antagonist or a PD-L1 antagonist) or the cancer or tumor of the subject progresses after an initial response to immune checkpoint inhibition (e.g., any immune checkpoint inhibitor described herein, such as a PD-1 antagonist or a PD-L1 antagonist).

[0823] In various embodiments, one or more additional therapies may comprise an antibody or an antigen-binding fragment thereof. Within this definition, immune checkpoint inhibitors include bispecific antibodies and immune cell-binding multivalent antibodies / fusion proteins / compositions known in the art. In some embodiments, one or more additional therapies comprising a bispecific antibody may comprise a bispecific antibody that binds to the same epitope of an immune checkpoint molecule, two different epitopes of the same immune checkpoint molecule, or different epitopes of two different immune checkpoints.

[0824] Those skilled in the art may implement various bispecific antibody formulations known in the art to target one or more of CTLA4, PD1, PD-L1 TIM-3, LAG-3, various B-7 ligands, B7H3, B7H4, CHK 1 and CHK2 kinases, BTLA, A2aR, OX40, 41BB, LIGHT, CD40, GITR, TGF-beta, SIRP-alpha, TIGIT, VSIG8, SIGLEC7, SIGLEC9, ICOS, FAS, BTNL2 and others for use in combinations described herein.

[0825] In various embodiments, one or more additional therapies may include immune cell-binding multivalent antibodies / fusion proteins / compositions.

[0826] In some embodiments of the present disclosure, one or more additional therapies are a population of immune cells that may be administered in combination with the non-polymorphic form, crystalline form, or crystalline salt form of Compound 1 in co-administration with a checkpoint inhibitor to treat a subject with cancer. In some embodiments, the immunotherapeutic agent is a population of immune cells, such as leukocytes (nucleated leukocytes), that contain (e.g., express) a receptor that binds to an antigen of interest. The leukocytes of the present disclosure may be, for example, neutrophils, eosinophils, basophils, lymphocytes, or monocytes. In some embodiments, the leukocytes are lymphocytes. Examples of lymphocytes include T cells, B cells, natural killer (NK) cells, or NKT cells. In some embodiments, the T cells are CD4+ Th (T helper) cells, CD8+ cytotoxic T cells, γδT cells, or regulatory (suppressor) T cells. In some embodiments, the immune cells are dendritic cells.

[0827] In some embodiments, the immune cells of the present disclosure are genetically engineered to express antigen-binding receptors. If the cells contain engineered (exogenous) nucleic acids, the cells are considered "engineered." The engineered nucleic acids of the present disclosure may be introduced into the cells by any known (e.g., conventional) method. For example, engineered nucleic acids can be transfected by electroporation (e.g., see [Heiser WC Transcription Factor Protocols: Methods in Molecular Biology.TM. 2000; 130: 117-134]), chemistry (e.g., calcium phosphate or lipids), transfection (e.g., see [Lewis WH, et al., Somatic Cell Genet. 1980 May; 6(3): 333-47; Chen C., et al., Mol Cell Biol. 1987 August; 7(8): 2745-2752]), fusion with bacterial protoplasts containing recombinant plasmids (e.g., see [Schaffner W. Proc Natl Acad Sci USA. 1980 April; 77(4): 2163-7]), direct microinjection of purified DNA into the cell nucleus (e.g., see [Capecchi MR Cell. 1980 November; It can be introduced into cells by [see 22(2 Pt 2): 479-88] or by retroviral delivery.

[0828] Some aspects of the present disclosure comprising one or more additional therapies provide an “adoptive cell” approach, which comprises isolating immune cells (e.g., T-cells) from a subject with cancer, genetically modifying the immune cells (e.g. to express an antigen-binding receptor such as a chimeric antigen receptor), expanding the cells in vitro, and then reintroducing the immune cells into the subject. This method generates a greater number of modified immune cells in the subject compared to what can be achieved by conventional gene transfer and vaccination methods. In some embodiments, the immune cells are isolated from the subject, expanded in vitro without genetic modification, and then reintroduced into the subject.

[0829] The immune cells of the present disclosure comprise receptors that bind to antigens such as antigens encoded by nucleic acids delivered exogenously as provided herein. In some embodiments, leukocytes are modified (e.g., genetically modified) to express receptors that bind to antigens. In some embodiments, the receptors may be naturally occurring antigen receptors (generally expressed in immune cells), recombinant antigen receptors (generally not expressed in immune cells), or chimeric antigen receptors (CARs). Naturally occurring and recombinant antigen receptors included in the present disclosure include T cell receptors, B cell receptors, NK cell receptors, NKT cell receptors, and dendritic cell receptors. "Chimeric antigen receptor" refers to an artificial immune cell receptor engineered to recognize and bind to an antigen expressed by a tumor cell. Generally, CARs are designed for T cells and are chimeras of the signaling domain of a T-cell receptor (TcR) complex and an antigen-recognition domain (e.g., a single-strand fragment of an antibody (scFv)) (Enblad et al., Human Gene Therapy. 2015; 26(8): 498-505, the entirety of which is incorporated herein by reference).

[0830] In some embodiments, the antigen-coupled receptor is a chimeric antigen receptor (CAR). T cells expressing a CAR are referred to as "CAR T cells." In some embodiments, the CAR T cell receptor comprises a signaling domain and an antigen-recognition domain of a T-cell receptor (TcR) complex (e.g., a single-chain fragment (scFv) of an antibody) (Enblad et al., Human Gene Therapy. 2015; 26(8): 498-505, the entirety of which is incorporated herein by reference).

[0831] There are four generations of CARs, each containing different components. The first generation CAR is an antibody-derived scFv linked to the CD3 zeta (zeta or z) intracellular signaling domain of the T-cell receptor through hinge and transmembrane domains. The second generation CAR is an additional domain, e.g., CD28, 4-1BB (41BB), or ICOS, incorporated to deliver co-stimulatory signals. The third generation CAR contains two co-stimulatory domains fused to the TcR CD3-zeta chain. The third generation co-stimulatory domain may include, e.g., a combination of CD3z, CD27, CD28, 4-1BB, ICOS, or OX40. In some embodiments, the CAR comprises an ectodomain (e.g., CD3) generally derived from a single-chain variable fragment (scFv), a hinge, a transmembrane domain, and an endodomain having a 1 (1st generation), 2 (2nd generation), or 3 (3rd generation) signaling domain derived from CD3Z and / or a co-stimulatory molecule (Maude et al., Blood. 2015; 125(26): 4017-4023; Kakarla and Gottschalk, Cancer J. 2014; 20(2): 151-155, the entire disclosure of which is incorporated herein by reference).

[0832] In some embodiments, the chimeric antigen receptor (CAR) is a T-cell redirected for universal cytokine killing (TRUCK), also known as a fourth-generation CAR. The TRUCK is a CAR-redirected T-cell used as a vehicle to produce and release transgenic cytokines that accumulate in targeted tissues, e.g., targeted tumor tissues. Transgenic cytokines are released when the target binds to the CAR. TRUCK cells can accumulate various therapeutic cytokines at the target. This results in therapeutic concentrations at the targeted site and allows systemic toxicity to be avoided.

[0833] CARs typically differ in their functional characteristics. The CD3 zeta signaling domain of the T-cell receptor activates and induces T-cell proliferation upon binding, but it can also induce anergy (resulting in the direct induction of peripheral lymphocyte tolerance due to a lack of response by the body's defense mechanisms). Lymphocytes are considered unresponsive when they do not react to a specific antigen. Adding a co-stimulatory domain to second-generation CARs improves the replication ability and persistence of modified T-cells. While similar antitumor effects are observed in vitro with CD28 or 4-1BB CARs, in vivo preclinical studies suggest that 4-1BB CARs can generate superior proliferation and / or persistence. Clinical trials suggest that while all of these second-generation CARs can induce significant T-cell proliferation in vivo, CARs containing the 4-1BB co-stimulatory domain exhibit longer persistence. Third-generation CARs enhance efficacy by combining multiple signaling domains (co-stimulators). Fourth-generation CARs are further modified with constitutive or inducible expression cassettes for transgenic cytokines released by CAR T-cells to modulate T-cell responses. For example, refer to the literature in which the entire disclosure is incorporated herein by reference [Enblad et al., Human Gene Therapy. 2015; 26(8): 498-505; Chmielewski and Hinrich, Expert Opinion on Biological Therapy. 2015; 15(8): 1145-1154].

[0834] In some embodiments, one or more additional therapies are first-generation chimeric antigen receptor CARs. In some embodiments, the chimeric antigen receptor is a second-generation CAR. In some embodiments, the chimeric antigen receptor is a third-generation CAR. In some embodiments, the chimeric antigen receptor is a fourth-generation CAR or a T-cell (TRUCK) redirected for universal cytokine killing.

[0835] In some embodiments, the chimeric antigen receptor (CAR) comprises an extracellular domain containing an antigen-binding domain, a transmembrane domain, and a cytoplasmic domain. In some embodiments, the CAR is fully human. In some embodiments, the antigen-binding domain of the CAR is specific to one or more antigens. In some embodiments, a “spacer” domain or a “hinge” domain is located between the extracellular domain (including the antigen-binding domain) of the CAR and the transmembrane domain, or between the cytoplasmic domain and the transmembrane domain of the CAR. The “spacer domain” refers to any oligopeptide or polypeptide that functions to connect the transmembrane domain to the extracellular domain and / or cytoplasmic domain in the polypeptide chain. The “hinge domain” refers to any oligopeptide or polypeptide that functions to provide flexibility to the CAR or its domains or to prevent steric hindrance of the CAR or its domains. In some embodiments, the spacer domain or hinge domain may contain up to 300 amino acids (e.g., 10 to 100 amino acids or 5 to 20 amino acids). In some embodiments, one or more spacer domain(s) may be included in other regions of the CAR.

[0836] In some embodiments, the CAR of the present disclosure comprises an antigen-binding domain, such as a single-stranded Fv (scFv), specific to a tumor antigen. The selection of the binding domain depends on the type and number of ligands defining the surface of the target cell. For example, the antigen-binding domain may be selected to recognize ligands that act as cell surface markers on target cells associated with specific disease states, such as cancer or autoimmune diseases. Thus, examples of cell surface markers that may act as ligands for the antigen-binding domain in the CAR of the present disclosure include those associated with cancer cells and / or other forms of disease cells. In some embodiments, the CAR is engineered to target a tumor antigen of interest by engineering a desired antigen-binding domain that specifically binds to an antigen on a tumor cell encoded by an engineered nucleic acid as provided herein.

[0837] An antigen-binding domain (e.g., scFv) that "specifically binds" to a target or epitope is a term understood in the art, and methods for determining such specific binding are also known in the art. A molecule is said to exhibit "specific binding" when it responds to or associates with a specific target antigen more frequently, more rapidly, for a longer period, and / or with greater affinity than with an alternative target. An antigen-binding domain (e.g., scFv) that specifically binds to a first target antigen may or may not specifically bind to a second target antigen. As such, "specific binding" does not necessarily require (or may include) exclusive binding.

[0838] In some embodiments, immune cells expressing the CAR are genetically modified to recognize multiple targets or antigens, which enables them to recognize unique target or antigen expression patterns in tumor cells. Examples of CARs capable of binding to multiple targets include: a "split signal CAR" that limits full immune cell activation to tumors expressing multiple antigens; a "tandem CAR" (TanCAR) comprising an ectodomain having two scFvs; and a "universal ectodomain CAR" incorporating an avidin or fluorescein isothiocyanate (FITC)-specific scFv to recognize tumor cells incubated with a tagged monoclonal antibody (Mab).

[0839] A CAR is considered "bispecific" if it recognizes two distinct antigens (having two distinct antigen recognition domains). In some embodiments, a bispecific CAR consists of two distinct antigen recognition domains existing in tandem on a single transgenic receptor (referred to as TanCAR; see, for example, the literature incorporated herein by reference in its entirety [Grada Z et al. Molecular Therapy Nucleic Acids 2013; 2: e105]). Accordingly, in some embodiments, the method comprises delivering to a tumor a combination comprising a non-polymorphic form, a crystalline form, or a crystalline salt form of compound 1 and an immunotherapeutic agent when co-administered with a checkpoint inhibitor, wherein the immunotherapeutic agent is a engineered nucleic acid encoding an antigen, or an engineered nucleic acid that induces the expression of a self-antigen, to be delivered to the tumor, and an immune cell expressing a bispecific CAR that binds to two antigens to be delivered to the tumor, one of which is encoded by the engineered nucleic acid.

[0840] In some embodiments, the CAR is an antigen-specific inhibitory CAR (iCAR) that can be used, for example, to avoid extratumor toxicity (Reference [Fedorov, VD et al. Sci. Transl. Med. published online Dec. 11, 2013], in its entirety incorporated herein by reference). The iCAR comprises, for example, an antigen-specific inhibitory receptor to block non-specific immunosuppression that may result from additional tumor target expression. The iCAR may be based, for example, on the inhibitory molecules CTLA-4 or PD-1. In some embodiments, these iCARs block T cell responses from endogenous T cell receptors or T cells activated by the activating CAR. In some embodiments, this inhibitory effect is transient.

[0841] In some embodiments, the CAR may be used for adoptive cell delivery, wherein the immune cells are removed from the subject and modified to express a receptor specific to an antigen, e.g., a tumor-specific antigen. Then, the modified immune cells capable of recognizing and killing cancer cells are reintroduced to the subject (references [Pule, et al., Cytotherapy. 2003; 5(3): 211-226; Maude et al., Blood. 2015; 125(26): 4017-4023], each of which is incorporated herein by reference in its entirety).

[0842] According to another aspect of the present disclosure, the tumor antigenic component in the vaccine of the present invention is any natural or synthetic tumor-associated protein or peptide, or a combination of tumor-associated protein and / or peptide or glycoprotein or glycopeptide. In another aspect, the antigenic component may be patient-specific or common to a number or majority of patients with a specific type of cancer. According to one aspect, the antigenic component consists of a cell lysate derived from tumor tissue removed from a patient being treated. In another aspect, the lysate may be fabricated or synthesized from exosomes derived from tumor tissue. In another aspect, the antigenic component consists of a cell lysate derived from tumor tissue or tumor cell lines extracted from one or more unrelated individuals.

[0843] In various embodiments, the tumor-associated antigen components of the vaccine may be prepared by any various well-known techniques. For individual protein components, the antigen proteins are isolated from tumor tissue or tumor cell lines by standard chromatographic means, such as high-pressure liquid chromatography or affinity chromatography, or alternatively synthesized by standard recombinant DNA techniques in suitable expression systems, such as E. coli, yeast, or plants. The tumor-associated antigen proteins are then purified from the expression system by standard chromatographic means. For peptide antigenic components, they are generally prepared by standard automated synthesis. Proteins and peptides may be modified by the addition of amino acids, lipids, and other agents to improve incorporation into the vaccine delivery system (e.g., multilayer liposomes). For tumor-associated antigenic components derived from the patient's own tumor or from tumors or cell lines from other individuals, the tumor tissue or single-cell suspension derived from the tumor tissue is typically homogenized in a suitable buffer. The homogenate may be fractionated, for example, by centrifugation, to isolate specific cellular components, such as cell membranes or soluble substances. Tumor material may be used directly or may be extracted to incorporate tumor-associated antigens into a vaccine using a buffer containing a low concentration of a suitable agent, such as a detergent. An example of a detergent suitable for extracting antigen proteins from tumor tissue, tumor cells, and tumor-cell membranes is diheptanoyl phosphatidylcholine. Exosomes derived from tumor tissue or tumor cells may be used as starting materials for the extraction of antigenic components or tumor-associated antigens to be incorporated into a vaccine, whether autologous or heterologous for the patient.

[0844] In some embodiments of the present disclosure, one or more additional therapies are cancer vaccine immunotherapies co-administered with a checkpoint inhibitor. In various examples, the cancer vaccine comprises at least one tumor-associated antigen, at least one immunostimulator, and optionally at least one cell-based immunotherapie. In some embodiments, the immunostimulator component in the cancer vaccine of the present disclosure is any Biological Response Modifier (BRM) having the ability to enhance the efficacy of the therapeutic cancer vaccine in inducing humoral and cellular immune responses against cancer cells in a patient. According to one aspect, the immunostimulator is a cytokine or a combination of cytokines. Examples of such cytokines include interferons, e.g., IFN-gamma; interleukins, e.g., IL-2, IL-15, and IL-23; colony-stimulating factors, e.g., M-CSF and GM-CSF; and tumor necrosis factor. According to another aspect, the immunostimulator component of the disclosed cancer vaccine comprises one or more co-modulatory agents, such as APC Toll-like receptor agonists or co-stimulatory / cell adhesion membrane proteins, which may or may not include immunomodulatory cytokines. Examples of Toll-like receptor agonists include lipids A and CpG and co-stimulatory / adhesion proteins, e.g., CD80, CD86, and ICAM-1.

[0845] In some embodiments, one or more additional therapies are immunostimulators selected from the group consisting of IFN-gamma (IFN-γ), IL-2, IL-15, IL-23, M-CSF, GM-CSF, tumor necrosis factor, lipid A, CpG, CD80, CD86, and ICAM-1, or combinations thereof. According to other embodiments, cell-based immunotherapies are selected from the group consisting of dendritic cells, tumor-infiltrating T lymphocytes, chimeric antigen receptor-modified T effector cells for the patient's tumor type, B lymphocytes, natural killer cells, myeloid cells, and any other cells of the patient's immune system, or combinations thereof. In one embodiment, the cancer vaccine immunostimulator comprises one or more cytokines, e.g., interleukin 2 (IL-2), GM-CSF, M-CSF, and interferon-gamma (IFN-γ), one or more Toll-like receptor agonists and / or adjuvants, e.g., monophosphoryl lipid A, lipid A, muramyl dipeptide (MDP) lipid conjugate, and double-stranded RNA, or one or more co-stimulating membrane proteins and / or cell adhesion proteins, e.g., CD80, CD86, and ICAM-1, or any combination of the above. In one embodiment, the cancer vaccine comprises an immunostimulator that is a cytokine selected from the group consisting of interleukin 2 (IL-2), GM-CSF, M-CSF, and interferon-gamma (IFN-γ). In another embodiment, the cancer vaccine comprises an immunostimulator that is an adjuvant selected from the group consisting of a Toll-like receptor agonist and / or monophosphoryl lipid A, lipid A and muramyl dipeptide (MDP) lipid conjugates and double-stranded RNA. In another embodiment, the cancer vaccine comprises an immunostimulator that is a cell adhesion protein selected from the group consisting of a co-stimulating membrane protein and / or CD80, CD86 and ICAM-1.

[0846] In various embodiments, one or more additional therapies may comprise a cancer vaccine, wherein the cancer vaccine comprises any tumor antigen that can potentially be used to form a fusion protein according to the present invention, in particular the following: (a) cancer-testis antigens comprising RAGE, BAGE, GAGE, and MAGE family polypeptides, e.g., GAGE-1, GAGE-2, MAGE-3, MAGE-4, MAGE-5, MAGE-6, and MAGE-12, as well as NY-ESO-1, SSX2, and SCP1 that can be used to resolve melanoma, lung, head and neck, NSCLC, breast, gastrointestinal, and bladder tumors; (b) p53 associated with various solid tumors, e.g., colorectal cancer, lung cancer, and head and neck cancer; p21 / Ras associated with melanoma, pancreatic cancer, and colorectal cancer; e.g., CDK4 associated with melanoma; e.g., MUM1 associated with melanoma; For example, cASPEC-8 associated with head and neck cancer; for example, CIA-0205 associated with bladder cancer; for example, HLA-A2-R1701, beta-catenin associated with melanoma; for example, TCR associated with T-cell non-Hodgkin lymphoma; for example, BCR-abl associated with chronic myeloid leukemia; triose phosphate isomerase; KIA-0205; mutated antigens including CDC-27 and LDLR-FUT; (c) for example, galectin 4 associated with colorectal cancer; for example, galectin 9 associated with Hodgkin's disease; for example, proteinase 3 associated with chronic myeloid leukemia; for example, WT-1 associated with various leukemias; for example, carbonic anhydrase associated with renal cancer; for example, aldolase A associated with lung cancer; for example, PRAME associated with melanoma; For example, HER-2 / neu associated with breast, colon, lung, and ovarian cancer; for example, mammaglobin and alpha-fetoprotein associated with liver cancer; for example, KSA associated with colorectal cancer; for example, gastrin associated with pancreatic and gastric cancer;For example, telomerase catalytic protein, MUC-1 associated with breast and ovarian cancer; for example, G-250 associated with renal cell carcinoma; for example, p53 associated with breast and colon cancer; and overexpressed antigens including carcinoembryonic antigens associated with breast, lung, and gastrointestinal cancers, e.g., colorectal cancer; (d) melanoma-melanocyte differentiation antigens such as MART-1 / Melan A; gpl00; MC1R; melanocyte-stimulating hormone receptor; tyrosinase; shared antigens including, for example, tyrosinase-associated protein-1 / TRP1 and tyrosinase-associated protein-2 / TRP2 associated with melanoma; (e) prostate-associated antigens including, for example, PAP, PSA, PSMA, PSH-P1, PSM-P1, and PSM-P2 associated with prostate cancer;(f) Immunoglobulin idiotypes associated with myeloma and B-cell lymphoma. In a given embodiment, one or more TAAs are pi 5, Hom / Mel-40, H-Ras, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR, Epstein-Barr virus antigen, EBNA, human papillomavirus (HPV) antigen including E6 and E7, hepatitis B and hepatitis C virus antigens, human T-cell lymphotropic virus antigen, TSP-180, pl85erbB2, pl80erbB-3, c-met, mn-23H1, TAG-72-4, CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, pi 6, TAGE, PSCA, CT7, 43-9F, 5T4, 791 Tgp72, beta-HCG, BCA225, BTAA, CA 125, CA It may be selected from 15-3(CA 27.29\BCAA), CA 195, CA 242, CA-50, CAM43, CD68\KP1, CO-029, FGF-5, Ga733(EpCAM), HTgp-175, M344, MA-50, MG7-Ag, MOV18, NB / 70K, NY-CO-1, RCAS1, SDCCAG16, TA-90(Mac-2 binding protein / cyclopilin C-related protein), TAAL6, TAG72, TLP, TPS, or any combination thereof.;

[0847] In some embodiments, one or more additional therapies may include a tumor antigen comprising the entire amino acid sequence of a human protein, part thereof, or a specific immunogenic epitope.

[0848] In various embodiments, one or more additional therapies may comprise mRNA operable to encode one or more of any of the aforementioned cancer antigens useful for synthesizing a cancer vaccine. In some exemplary embodiments, the mRNA-based cancer vaccine may have one or more of the following characteristics: a) the mRNA encoding each cancer antigen is interspersed by cleavage-sensitive sites; b) the mRNA encoding each cancer antigen is directly linked to each other without a linker; c) the mRNA encoding each cancer antigen is linked to each other by a single nucleotide linker; d) each cancer antigen comprises 20 to 40 amino acids and contains a centrally located SNP mutation; e) at least 40% of the cancer antigens have the highest affinity for class I MHC molecules from the subject; f) at least 40% of the cancer antigens have the highest affinity for class II MHC molecules from the subject; g) at least 40% of the cancer antigens have a predicted binding affinity of IC > 500 nM for HLA-A, HLA-B, and / or DRB1; h) mRNA encodes 1 to 15 cancer antigens; i) 10% to 60% of the cancer antigens have binding affinities for class I MHC, and 10% to 60% of the cancer antigens have binding affinities for class II MHC; and / or j) mRNA encoding cancer antigens is arranged so that the cancer antigens are aligned to minimize pseudo-epitopes.

[0849] In various embodiments, one or more additional therapies comprise at least one RNA polynucleotide having an open reading frame encoding at least one antigenic polypeptide or an immunogenic fragment thereof, and an RNA vaccine that induces an immune response specific to the antigenic polypeptide or its immunogenic fragment in a subject by administering the same composition or a separate composition in simultaneous or sequential doses, wherein the anti-antigenic polypeptide antibody titer in the subject increases after vaccination compared to the anti-antigenic polypeptide antibody titer in a subject vaccinated with a traditional vaccine of a prophylactic effective dose against cancer. The "anti-antigenic polypeptide antibody" is a serum antibody that specifically binds to the antigenic polypeptide.

[0850] The prophylactic effective dose is a therapeutically effective dose that prevents the progression of cancer at a clinically acceptable level. In some embodiments, the therapeutically effective dose is the dose listed in the package insert for the vaccine. As used herein, the term "traditional vaccine" refers to a vaccine other than the mRNA vaccine of the present invention. For example, traditional vaccines include, but are not limited to, live microbial vaccines, inactivated microbial vaccines, subunit vaccines, protein antigen vaccines, DNA vaccines, etc. In exemplary embodiments, the traditional vaccine is a vaccine that has obtained regulatory approval and / or has been registered by a national drug regulatory agency, e.g., the U.S. Food and Drug Administration (FDA) or the European Medicines Agency (EMA).

[0851] In some embodiments, the anti-antigenic polypeptide antibody titer in a subject increases by 1 log to 10 log after vaccination compared to the anti-antigenic polypeptide antibody titer in a subject vaccinated with a prophylactic effective dose of a traditional vaccine for cancer. In some embodiments, the anti-antigenic polypeptide antibody titer in a subject increases by 1 log after vaccination compared to the anti-antigenic polypeptide antibody titer in a subject vaccinated with a prophylactic effective dose of a traditional vaccine for cancer. In some embodiments, the anti-antigenic polypeptide antibody titer in a subject increases by 2 log after vaccination compared to the anti-antigenic polypeptide antibody titer in a subject vaccinated with a prophylactic effective dose of a traditional vaccine for cancer.

[0852] An aspect of the present invention provides a nucleic acid vaccine comprising one or more RNA polynucleotides having an open reading frame encoding a first antigenic polypeptide, wherein the RNA polynucleotides are present in a formulation for in vivo administration to a host, which imparts an antibody titer superior to the seroprotective standard for the first antigen for an acceptable percentage of human subjects. In some embodiments, the antibody titer produced by the mRNA vaccine of the present invention is a neutralizing antibody titer. In some embodiments, the neutralizing antibody titer is greater than that of the protein vaccine. In other embodiments, the neutralizing antibody titer produced by the mRNA vaccine of the present invention is greater than that of the auxiliary protein vaccine. In another embodiment, the neutralizing antibody titer generated by the mRNA vaccine of the present invention is 1,000 to 10,000, 1,200 to 10,000, 1,400 to 10,000, 1,500 to 10,000, 1,000 to 5,000, 1,000 to 4,000, 1,800 to 10,000, 2,000 to 10,000, 2,000 to 5,000, 2,000 to 3,000, 2,000 to 4,000, 3,000 to 5,000, 3,000 to 4,000, or 2,000 to 2,500. The neutralizing titer is expressed as the highest serum dilution required to achieve a 50% reduction in the number of plaques.

[0853] In a preferred embodiment, the RNA vaccine immunotherapeutic agent of the present disclosure (e.g., mRNA vaccine) produces a level, concentration, and / or titer of antigen-specific antibodies in the blood or serum of a vaccinated subject that is prophylactic and / or therapeutically effective. As defined herein, the term antibody titer refers to the amount of antigen-specific antibodies produced in a subject, e.g., a human subject. In exemplary embodiments, the antibody titer is expressed as the reciprocal of the maximum dilution (sequential dilution) that still yields a positive result. In exemplary embodiments, the antibody titer is determined or measured by an enzyme-linked immunosorbent assay (ELISA). In exemplary embodiments, the antibody titer is determined or measured by a neutralization assay, e.g., a micro-neutralization assay. In certain embodiments, the antibody titer measurement is expressed as a ratio, such as 1:40, 1:100, etc.

[0854] In an exemplary embodiment of the present invention, the effective vaccine produces antibody titers of 1:40 or greater, 1:100 or greater, 1:400 or greater, 1:1000 or greater, 1:2000 or greater, 1:3000 or greater, 1:4000 or greater, 1:500 or greater, 1:6000 or greater, 1:7500 or greater, or 1:10000 or greater. In an exemplary embodiment, antibody titers are produced or reached 10 days after vaccination, 20 days after vaccination, 30 days after vaccination, 40 days after vaccination, or 50 days or more after vaccination. In an exemplary embodiment, titers are produced or reached after a single dose of the vaccine administered to the subject. In another embodiment, titers are produced or reached after multiple doses, for example, after a first dose and a second dose (e.g., a booster dose). In an exemplary embodiment of the present invention, antigen-specific antibodies are measured in units of g / mL or in IU / L (International Unit per liter) or mIU / mL (Milli International Unit per mL). In an exemplary embodiment of the present invention, an effective vaccine produces greater than 0.5 μg / mL, greater than 0.1 μg / mL, greater than 0.2 μg / mL, greater than 0.35 μg / mL, greater than 0.5 μg / mL, greater than 1 μg / mL, greater than 2 μg / mL, greater than 5 μg / mL, or greater than 10 μg / mL. In an exemplary embodiment of the present invention, an effective vaccine produces greater than 10 mIU / mL, greater than 20 mIU / mL, greater than 50 mIU / mL, greater than 100 mIU / mL, greater than 200 mIU / mL, greater than 500 mIU / mL, or greater than 1000 mIU / mL. In an exemplary embodiment, an antibody level or concentration is generated or reached 10 days after vaccination, 20 days after vaccination, 30 days after vaccination, 40 days after vaccination, or 50 days or more after vaccination. In an exemplary embodiment, a level or concentration is generated or reached after a single dose of the vaccine administered to the subject. In another embodiment, a level or concentration is generated or reached after multiple doses, for example, after a first dose and a second dose (e.g., a booster dose).In exemplary embodiments, antibody levels or concentrations are determined or measured by an enzyme-linked immunosorbent assay (ELISA). In exemplary embodiments, antibody levels or concentrations are determined or measured by a neutralization assay, e.g., a microneutralization assay. Additionally, a nucleic acid vaccine is provided comprising one or more RNA polynucleotides or concatemeric polypeptides having an open reading frame encoding a first antigenic polypeptide, wherein the RNA polynucleotides have a stabilizing element or are formulated with an adjuvant and are present in a formulation for in vivo administration to a host to induce high antibody titers that persist longer than the antibody titers induced by an mRNA vaccine encoding the first antigenic polypeptide. In some embodiments, the RNA polynucleotides are formulated to generate neutralizing antibodies within one week of a single administration. In some embodiments, the adjuvant is selected from cationic peptides and immunomodulatory nucleic acids. In some embodiments, the cationic peptide is a protamine.

[0855] The immunotherapeutic agent comprises a nucleic acid vaccine comprising one or more RNA polynucleotides having an open reading frame that includes at least one chemical modification or optionally does not include a nucleotide modification, an open reading frame or a chain polypeptide encoding a first antigenic polypeptide, wherein the RNA polynucleotide is present in a formulation for in vivo administration to a host such that the level of antigen expression in the host is significantly exceeded by the level of antigen expression produced by an mRNA vaccine that has a stabilizing element or is formulated with an adjuvant and encoding a first antigenic polypeptide.

[0856] Another embodiment provides a nucleic acid vaccine comprising one or more RNA polynucleotides having an open reading frame that includes at least one chemical modification or optionally does not include a nucleotide modification, an open reading frame encoding a first antigenic polypeptide, or a chain polypeptide, wherein the vaccine has at least 10 times less RNA polynucleotides than required for an unmodified mRNA vaccine to produce an equivalent antibody titer. In some embodiments, the RNA polynucleotides are present in a dosage of 25 to 100 micrograms.

[0857] Aspects of the present invention also provide a unit of use vaccine comprising 10 to 400 μg of at least one RNA polynucleotide having an open reading frame that comprises at least one chemical modification or optionally does not comprise a nucleotide modification, an open reading frame or chain polypeptide encoding a first antigenic polypeptide, and a pharmaceutically acceptable excipient, formulated for delivery to a human subject. In some embodiments, the vaccine further comprises cationic lipid nanoparticles.

[0858] An aspect of the present invention provides a method for generating, maintaining, or restoring antigenic memory for a tumor in an individual or population of individuals, comprising the step of administering said individual or population antigenic memory booster nucleic acid vaccine, said RNA polynucleotide comprising at least one RNA polynucleotide, said polynucleotide comprising at least one chemical modification or optionally not comprising a nucleotide modification, said RNA polynucleotide comprising two or more codon-optimized open reading frames, said open reading frames encoding a set of reference antigenic polypeptides, and (b) optionally a pharmaceutically acceptable excipient. In some embodiments, the vaccine is administered to the individual via a route selected from the group consisting of intramuscular administration, intradermal administration, and subcutaneous administration. In some embodiments, the administration step comprises the step of bringing the muscle tissue of the subject into contact with a device suitable for injection of the composition. In some embodiments, the administration step comprises the step of bringing the muscle tissue of the subject into contact with a device suitable for injection of the composition together with electroporation.

[0859] An aspect of the present invention provides a method for vaccinating a subject, comprising the step of administering a single dose of a nucleic acid vaccine comprising one or more RNA polynucleotides or chain polypeptides having an open reading frame encoding a first antigenic polypeptide, in an amount effective for vaccinating the subject, to the subject.

[0860] Another embodiment provides a nucleic acid vaccine comprising one or more RNA polynucleotides having an open reading frame comprising at least one chemical modification, an open reading frame encoding a first antigenic polypeptide, or a chain polypeptide, wherein the vaccine has at least 10 times less RNA polynucleotides than required for an unmodified mRNA vaccine to produce an equivalent antibody titer. In some embodiments, the RNA polynucleotides are present in a dosage of 25 to 100 micrograms.

[0861] In some embodiments, one or more additional therapies, a non-polymorphic form, a crystalline form, or a crystalline salt form of Compound 1 may be a bispecific antibody immunotherapeutic agent. The bispecific antibody may comprise a protein construct having a first antigen-binding moiety and a second antigen-binding site that binds to cytotoxic immune cells. The first antigen-binding site may bind to a tumor antigen specifically treated with the combination of the present invention. For example, the first antigen-binding moiety may bind to non-limiting examples of tumor antigens selected from the following: in particular EGFR, HGFR, Her2, Ep-CAM, CD20, CD30, CD33, CD47, CD52, CD133, CEA, gpA33, mucin, TAG-72, CIX, PSMA, folate-binding protein, GD2, GD3, GM2, VEGF. VEGFR, integrin αVβ3, integrin α5β1, MUC1, ERBB2, ERBB3, MET, IGF1R, EPHA3, TRAILR1, TRAILR2, RANKL, FAP, and tenasin. In some embodiments, the first antigen-binding moiety is specific to a protein or peptide overexpressed on tumor cells compared to a corresponding non-tumor cell. In some embodiments, the first antigen-binding moiety is specific to a protein overexpressed on tumor cells compared to a corresponding non-tumor cell. As used herein, "corresponding non-tumor cell" refers to a non-tumor cell that is of the same cell type as the tumor cell. It should be noted that such proteins are not necessarily different from the tumor antigen.Non-limiting examples include carcinoembryonic antigen (CEA), which is overexpressed in most colorectal, rectal, breast, lung, pancreatic, and gastrointestinal carcinomas; heregulin receptor (HER-2, neu, or c-erbB-2), which is commonly overexpressed in breast, ovarian, colorectal, lung, prostate, and cervical cancers; epidermal growth factor receptor (EGFR), which is highly expressed in various solid tumors including breast, head and neck, non-small cell lung, and prostate; asarloglycoprotein receptor; transferrin receptor; serpin enzyme complex receptor, which is expressed in hepatocytes; fibroblast growth factor receptor (FGFR), which is overexpressed in pancreatic ductal adenocarcinoma cells; vascular endothelial growth factor receptor (VEGFR) for anti-angiogenic gene therapy; folate receptor, which is selectively overexpressed in 90% of non-mucinous ovarian carcinomas; cell surface carbohydrate layer; carbohydrate receptor; and polymeric immunoglobulin receptor.

[0862] The second antigen-binding moiety is any molecule that specifically binds to an antigen, protein, or polypeptide expressed on the surface of a cytotoxic immune cell (CIK cell). Exemplary non-limiting antigens expressed on the surface of cytotoxic immune cells suitable for use with the present disclosure may include CD2, CD3, CD4, CD5, CD8, CD11a, CD11b, CD14, CD16a, CD27, CD28, CD45, CD45RA, CD56, CD62L, Fc receptor, LFA, LFA-1, TCRαβ, CCR7, macrophage inflammatory protein 1a, perforin, PD-1, PD-L1, PD-L2, or CTLA-4, LAG-3, OX40, 41BB, LIGHT, CD40, GITR, TGF-beta, TIM-3, SIRP-alpha, TIGIT, VSIG8, BTLA, SIGLEC7, SIGLEC9, ICOS, B7H3, B7H4, FAS, BTNL2, CD27, and Fas ligands. In some embodiments, the second antigen-binding moiety binds to CD3 of cytotoxic immune cells, e.g., CIK cells. In some embodiments, the second antigen-binding moiety binds to CD56 of cytotoxic immune cells. In some embodiments, the second antigen-binding moiety binds to the Fc receptor of cytotoxic immune cells. In some embodiments, the Fc region of a bispecific antibody binds to the Fc receptor of cytotoxic immune cells. In some embodiments, the second antigen-binding moiety is any molecule that specifically binds to an antigen expressed on the surface of a cytotoxic immune cell (e.g., CIK cell). The second antigen-binding moiety is specific to the antigen of the cytotoxic immune cell. Exemplary cytotoxic immune cells include, but are not limited to, CIK cells, T-cells, CD8+ T cells, activated T-cells, monocytes, natural killer (NK) cells, NK T cells, lymphokine-activated killer (LAK) cells, macrophages, and dendritic cells.The second antigen-binding moiety specifically binds to antigens expressed on the surface of cytotoxic immune cells. Exemplary non-limiting antigens expressed on the surface of cytotoxic immune cells suitable for modulation of the present disclosure may include CD2, CD3, CD4, CD5, CD8, CD11a, CD11b, CD14, CD16a, CD27, CD28, CD45, CD45RA, CD56, CD62L, Fc receptor, LFA, LFA-1, TCRαβ, CCR7, macrophage inflammatory protein 1a, perforin, PD-1, PD-L1, PD-L2, or CTLA-4, LAG-3, OX40, 41BB, LIGHT, CD40, GITR, TGF-beta, TIM-3, SIRP-alpha, TIGIT, VSIG8, BTLA, SIGLEC7, SIGLEC9, ICOS, B7H3, B7H4, FAS, BTNL2, CD27, and Fas ligands. In another embodiment, the bispecific antibody modulator is an activator of a co-stimulatory molecule (e.g., an OX40 agonist). In one embodiment, the OX40 agonist is a bispecific antibody molecule against OX40 and another tumor antigen or co-stimulatory antigen. The OX40 agonist may be administered alone or in combination with other immunomodulators, for example, in combination with inhibitors of PD-1, PD-L1, CTLA-4, CEACAM (e.g., CEACAM-1, -3 and / or -5), TIM-3, or LAG-3 (e.g., antibody constructs). In some embodiments, the anti-OX40 antibody molecule is a bispecific antibody that binds to GITR and PD-1, PD-L1, CTLA-4, CEACAM (e.g., CEACAM-1, -3 and / or -5), TIM-3, or LAG-3. In one exemplary embodiment, an OX40 antibody molecule is administered in combination with an anti-PD-1 antibody molecule (e.g., an anti-PD-1 molecule as described herein). The OX40 antibody molecule and the anti-PD-1 antibody molecule may be in the form of separate antibody compositions or bispecific antibody molecules.In another embodiment, the OX40 agonist may be administered in combination with an agonist of another co-stimulatory molecule, e.g., GITR, CD2, CD27, CD28, CDS, ICAM-1, LFA-1 (CD11a / CD18), ICOS (CD278), 4-1BB (CD137), CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160, B7-H3, or CD83 ligand. In some embodiments, the second antigen-binding moiety binds to an Fc receptor on cytotoxic immune cells, e.g., CIK cells.

[0863] In some embodiments, the bispecific antibody immunotherapeutic agent has specificity for tumor antigens and CIK cells, which brings tumor antigen-expressing tumor cells into proximity to CIK cells and induces the elimination of tumor cells through the anti-tumor cytotoxicity of CIK cells. In some embodiments, the bispecific antibody has specificity for tumor antigens but not for CIK cells, but the Fc region of the bispecific antibody can bind to the Fc receptor of CIK cells, which ultimately brings tumor cells into proximity to CIK cells and induces the elimination of tumor cells through the anti-tumor cytotoxicity of CIK cells. In some embodiments, the bispecific antibody has specificity for CIK cells but not for tumor cells, but the Fc region of the bispecific antibody can bind to the Fc receptor of tumor cells, which ultimately brings tumor cells into proximity to CIK cells and induces the elimination of tumor cells through the anti-tumor cytotoxicity of CIK cells.

[0864] In some embodiments, one or more additional therapies are immune cell-binding multivalent antibody / fusion protein / composition immunotherapies co-administered with a checkpoint inhibitor. In various embodiments, one or more additional therapies may comprise immune cell-binding multivalent antibody / fusion protein / compositions that may include any engineered antibody that does not mimic a recombinant structure, e.g., the original IgG structure. Here, different strategies are used to multiplex antibody fragments. For example, shortening the peptide linker between V domains causes scFv to self-associate into a dimer (diabody; 55 kDa). The bispecific diabody is formed by the non-covalent association of two VHA-VLB and VHB-VLA fragments expressed in the same cell. This leads to the formation of a heterodimer having two different binding sites. A single-stranded dibody (sc-diabody) is a bispecific molecule in which VHA-VLB and VHB-VLA fragments are linked together by an additional third linker. A tandem dibody (Tandab) is a quadrivalent bispecific antibody produced by two sc-diabodies.

[0865] Additionally, die-diebodies known in the art are included. This 130-kDa molecule is formed by the fusion of a diebody to the N-terminus of the CH3 domain of IgG to produce an IgG-like structure. Additional diebody derivatives are triabodies and tetrabodies that fold into trimeric and tetrameric fragments by shortening the linker to fewer than 5 or 0 to 2 residues. Also, the (scFv)2 construct known as the 'Bispecific T cell engaging antibody' (BITE) is exemplified. BITE is a bispecific single-stranded antibody consisting of two scFv antibody fragments linked via a flexible linker, which is directed against the surface antigen of a target cell and CD3 of a T cell. Also, divalent (Fab)2 and trivalent (Fab)3 antibody forms are exemplified. Additionally, minibodies and trimerbodies derived from scFv are exemplified. Exemplary constructs useful for targeting a target tumor antigen may include one or more of the following: diabody, single-stranded (sc)-diabody (scFv)2, miniantibody, minibody, Varnais-Vasta, scFv-Fc, sc(Fab)2, trimeric antibody construct, triabody antibody construct, trimerbody antibody construct, tribody antibody construct, collabody antibody construct, (scFv-TNFa)3, F(ab)3 / DNL. Exemplary cytotoxic immune cells include, but are not limited to, CIK cells, T-cells, CD8+ T cells, activated T-cells, monocytes, natural killer (NK) cells, NK T cells, lymphokine-activated killer (LAK) cells, macrophages, and dendritic cells.

[0866] In some embodiments, one or more additional therapies are radioconjugates.

[0867] In various embodiments, the radioconjugate is a small molecule or large molecule (referred herein to as "cell targeting agent"), e.g., polypeptide, antibody, or antibody fragment thereof, which is coupled to or otherwise attached to a radionuclide or a plurality of radionuclides such that binding to the target of the radioconjugate (a protein or molecule on or within a cancer cell) causes death or morbidity of said cancer cell. In various embodiments, the radioconjugate may be a cell targeting agent labeled with a radionuclide, or the cell targeting agent may be coupled to or otherwise attached to a particle, microparticle, or nanoparticle containing a plurality of radionuclides, wherein the radionuclides may be the same or different. A method for synthesizing the radioconjugate is known in the art and may include a class of immunoglobulin conjugated to a toxic radionuclide or an antigen-binding portion thereof.

[0868] In some embodiments, one or more additional therapies may be molecules that bind to cancer cells and may be known as “cell targeting agents.” As used herein, exemplary cell targeting agents may enable drug-containing nanoparticles or radionuclides to target specific types of cells of interest. Examples of cell targeting agents include, but are not limited to, small molecules (e.g., folates, adenosine, purines) and large molecules (e.g., peptides or antibodies) that bind to or target a target tumor-associated antigen. Examples of tumor-associated antigens include, but are not limited to, adenosine receptor, alpha v beta 3, aminopeptidase P, alpha-fetoprotein, cancer antigen 125, carcinoembryonic antigen, c-caveolin-1, chemokine receptor, clusterin, oncofetogenic antigen, CD20, epithelial tumor antigen, melanoma-associated antigen, Ras, p53, Her2 / Neu, ErbB2, ErbB3, ErbB4, folate receptor, prostate-specific membrane antigen, prostate-specific antigen, purine receptor, radiation-induced cell surface receptor, serpine B3, serpine B4, squamous cell carcinoma antigen, thrombospondin, tumor antigen 4, tumor-associated glycoprotein 72, tyrosinase, and tyrosine kinase. In some embodiments, the cell targeting agent is a folate or folate derivative that specifically binds to the folate receptor (FR). In some embodiments, the cell targeting agent is particularly EGFR, HGFR, Her2, Ep-CAM, CD20, CD30, CD33, CD47, CD52, CD133, CEA, gpA33, mucin, TAG-72, CIX, PSMA, folate-binding protein, GD2, GD3, GM2, VEGF.It is an antibody, a bispecific antibody, a trispecific antibody, or an antigen-binding construct thereof that specifically binds to a cancer antigen selected from VEGFR, integrin αVβ3, integrin α5β1, MUC1, ERBB2, ERBB3, MET, IGF1R, EPHA3, TRAILR1, TRAILR2, RANKL, FAP, and Tenasin.

[0869] The use of folates as targeting agents in radioconjugates also allows both tumor cells and regulatory T (Treg) cells to be targeted for destruction. It is well known that a large number of Treg cells suppress tumor immunity. Specifically, Treg cells suppress (foreign and self) reactive T cells without killing T cells through contact-dependent or cytokine secretion (e.g., IL-10, TGF-beta, etc.). FR4 is selectively upregulated in Treg cells. Antibody blockade of FR4 has been shown to deplete Treg cells and induce tumor immunity in tumor-bearing mice. Therefore, folate-coated PBM nanoparticles carrying cytotoxic agents will target FR-expressing cells for destruction, which will inhibit tumor progression directly (i.e., BrCa cells) and indirectly (i.e., breast tumor-associated and peripheral Treg cells).

[0870] In another additional embodiment, the targeting agent is an antibody or peptide or immune cell-binding multivalent antibody / fusion protein / composition capable of binding to a tumor-associated antigen, comprising but not limited to: adenosine receptor, alpha v beta 3, aminopeptidase P, alpha-fetoprotein, cancer antigen 125, carcinoembryonic antigen, caveolin-1, chemokine receptor, clusterin, oncofetogenic antigen, CD20, human growth factor receptor (HGFR), epithelial tumor antigen, melanoma-associated antigen, MUC1, Ras, p53, Her2 / Neu, ErbB2, ErbB3, ErbB4, folate receptor, prostate-specific membrane antigen, prostate-specific antigen, purine receptor, radiation-induced cell surface receptor, serpine B3, serpine B4, squamous cell carcinoma antigen, thrombospondin, tumor antigen 4, tumor-associated glycoprotein 72, tyrosinase, Tyrosine kinase, etc.

[0871] In some embodiments, one or more additional therapies are vaccination protocols. In some embodiments, vaccines may include those used to stimulate an immune response to cancer antigens.

[0872] The amounts of both the non-polymorphic form, crystalline form, or crystalline salt form of Compound 1 as disclosed herein, which may be combined with excipient materials to produce a single dosage form, and one or more additional therapeutic agents (among compositions containing additional therapeutic agents as described above) will vary depending on the host being treated and the specific mode of administration. In certain embodiments, the composition of the present invention is formulated so that a dosage of 0.01 to 100 mg / kg body weight / day of the present invention can be administered.

[0873] Additional therapeutic agents may act synergistically. Therefore, the amount of additional therapeutic agent in this composition may be less than required in monotherapy using only that therapeutic agent, or there may be fewer side effects in the patient when a lower dose is used. In certain embodiments, a dosage of 0.01 to 10,000 μg / kg body weight / day of additional therapeutic agent in this composition may be administered.

[0874] In some embodiments, one or more additional therapies are kinase inhibitors selected from the following: Akt1, Akt2, Akt3, TGF-βR, PKA, PKG, PKC, CaM-kinase, phosphorylase kinase, MEKK, ERK, MAPK, mTOR, EGFR, HER2, HER3, HER4, 1NS-R, IGF-1R, IR-R, PDGFαR, PDGFβ / R, CSFIR, KIT, FLK-II, KDR / FLK-1, FLK-4, flt-1, FGFR1, FGFR2, FGFR3, FGFR4, Ron, Sea, TRKA, TRKB, TRKC, FLT3, VEGFR / Flt2, Flt4, EphAl, EphA2, EphA3, EphB2, EphB4, Tie2, Src, Fyn, Lck, Fgr, Btk, Fak, SYR, FRK, JAK, ABL, ALK, CDK7, CDK12, CDK13, KRAS, and B-Raf. In some embodiments, one or more additional therapies are inhibitors of CD47 and MALT1 proteins.

[0875] In some embodiments, one or more additional therapies are poly ADP ribose polymerase (PARP) inhibitors. Exemplary PARP inhibitors include, but are not limited to, olaparib (Lynparza®), rucaprib (Rubraca®), niraparib (Zejula®), talzoparib (Talzenna®), and TPST-1120.

[0876] In some embodiments, one or more additional therapies are kinase inhibitors. Exemplary kinase inhibitors include imatinib, baricitinib, gefitinib, erlotinib, sorafenib, dasatinib, sunitinib, lapatinib, nilotinib, pirfenidone, zanubrutinib, upadacitinib, fedratinib, entrectinib, alpelisib, pazopanib, crizotinib, vemurafenib, vandetanib, ruxolitinib, axitinib, bosutinib, regorafenib, tofacitinib, cabozantinib, ponatinib, trametinib, dabrafenib, afatinib, ibrutinib, ceritinib, idelalisib, nintedanib, palbociclib, lenvatinib, cobimetinib, abemaciclib, acalabrutinib, alectinib, binimetinib, brigatinib, encorafenib, erdafitinib. Includes everolimus, fostamatinib, gilter, larotrectinib, rolatinib, netasudil, osimertinib, pexidatinib, ribociclib, temsirolimus, XL-147, XL-765, XL-499, and XL-880. In some embodiments, the kinase inhibitor is an HSP90 inhibitor (e.g., XL888), a liver X receptor (LXR) modulator, a retinoid-related orphan receptor gamma (RORy) modulator, a CK1 inhibitor, a CKl-α inhibitor, a Wnt pathway inhibitor (e.g., SST-215), or an inorganic corticosteroid receptor inhibitor (e.g., esaxerenone or XL-550) for the treatment of diseases disclosed herein, such as cancer.

[0877] In some embodiments, one or more additional therapies are polatuzumab vedotin.

[0878] A pharmaceutical composition comprising a non-polymorphic form, a crystalline form, or a crystalline salt form of Compound 1 according to the present disclosure will typically comprise an active amount of a non-polymorphic form, a crystalline form, or a crystalline salt form of Compound 1 dispersed in a pharmaceutically acceptable excipient, an immunotherapeutic agent, and / or both. The phrase “pharmaceutically or pharmacologically acceptable” refers to molecular entities and compositions that do not cause adverse, allergic, or other undesirable reactions when appropriately administered to animals, such as humans, for example. The preparation of a pharmaceutical composition comprising a non-polymorphic form, a crystalline form, or a crystalline salt form of Compound 1 is described in the literature [Remington's Pharmaceutical Sciences, 21 st As exemplified by Ed., (Lippincott, Williams and Wilkins Philadelphia, PA, 2006), it will be known to those skilled in the art in light of the present disclosure. Furthermore, it will be understood that for administration to animals (e.g., humans), the formulation must meet standards of sterility, pyrogenity, general safety, and purity. Specific examples of pharmacologically acceptable excipients for combination compositions comprising a non-polymorphic form, a crystalline form, or a crystalline salt form of Compound 1 mixed with an immunotherapeutic agent as described herein are borate buffer or sterile saline solution (0.9% NaCl).

[0879] The formulations of immunotherapies used according to the present disclosure, e.g., immune checkpoint modulator antibodies, are in the form of lyophilized formulations or aqueous solutions and / or suspensions [Remington's Pharmaceutical Sciences 21 stAs fully described and exemplified in Ed., (Lippincott, Williams and Wilkins Philadelphia, PA, 2006), antibodies having a desired degree of purity may be prepared for storage by mixing them with selectively pharmaceutically acceptable excipients or stabilizers. Acceptable excipients, buffers, or stabilizers are non-toxic to the recipient at the doses and concentrations used and include suitable aqueous and / or non-aqueous excipients that can be used in the pharmaceutical compositions of the present disclosure, e.g., water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils, e.g., olive oil, and injectable organic esters, e.g., ethyl oleate. Suitable fluidity may be maintained by the use of coating materials, e.g., lecithin, maintaining the necessary particle size in the case of dispersions, and the use of surfactants, buffers, e.g., phosphates, citrates, and other organic acids. Antioxidants, for example, (1) water-soluble antioxidants, e.g., ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; (2) oil-soluble antioxidants, e.g., ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, etc.; and (3) metal chelating agents, e.g., citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.; Preservatives (e.g., octade-syldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens, e.g., methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); and low molecular weight (less than about 10 residues) may be included.Other exemplary pharmaceutically acceptable excipients may include polypeptides; proteins, e.g., serum albumin, gelatin, or immunoglobulin; hydrophilic polymers, e.g., polyvinylpyrrolidone; amino acids, e.g., glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrin; chelating agents, e.g., EDTA; sugars, e.g., sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions, e.g., sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants, e.g., TWEEN™, PLURONICS™, or polyethylene glycol (PEG).

[0880] In one exemplary embodiment, the pharmaceutical composition may optionally include pH adjusters, buffers, and toxicity modifiers, pharmaceutically acceptable auxiliary substances required for approximate physiological conditions, such as sodium acetate, sodium chloride, potassium chloride, calcium chloride, and sodium lactate. In some embodiments, the checkpoint inhibitor antibody of the present disclosure or its antigen-binding fragment may be formulated, lyophilized for storage, and reconstituted in a suitable excipient before use according to lyophilization and reconstitution techniques known in the art. In one exemplary pharmaceutical composition comprising one or more checkpoint inhibitor antibodies or their antigen-binding fragments, the composition is formulated as a sterile, preservative-free solution of one or more checkpoint inhibitor antibodies or their antigen-binding fragments for intravenous or subcutaneous administration. The formulation may be supplied as a single-use, pre-filled pen, for example, a pre-filled glass syringe of about 1 ml, or as a single-use institutional vial. Preferably, the pharmaceutical composition comprising a checkpoint inhibitor antibody or an antigen-binding fragment thereof is clear and colorless and has a pH of about 6.9 to 5.0, preferably 6.5 to 5.0, and much more preferably about 6.0 to about 5.0. In various embodiments, a formulation comprising the pharmaceutical composition may contain about 500 mg to about 10 mg, or about 400 mg to about 20 mg, or about 300 mg to about 30 mg, or about 200 mg to about 50 mg of a checkpoint inhibitor antibody or an antigen-binding fragment thereof per ml of solution when reconstituted and administered to a subject. Exemplary injectable or infusion excipients may include mannitol, citrate monohydrate, dibasic sodium phosphate dihydrate, monobasic sodium phosphate dihydrate, polysorbate 80, sodium chloride, sodium citrate, and water in the case of parenteral administration, e.g., intravenously, intramuscularly, intraperitoneally, or subcutaneously.

[0881] In another exemplary embodiment, one or more immunotherapeutic agents or antigen-binding fragments thereof are formulated for intravenous or subcutaneous administration in a sterile aqueous solution containing 1 mg / ml to 75 mg / ml, or more preferably about 5 mg / ml to 60 mg / ml, or much more preferably about 10 mg / ml to 50 mg / ml, or much more preferably about 10 mg / ml to 40 mg / ml of antibodies, together with sodium acetate, polysorbate 80, and sodium chloride (pH in the range of about 5 to 6). Preferably, the intravenous or subcutaneous formulation is a sterile aqueous solution containing 5 mg / ml, 10 mg / ml, 15 mg / ml, 20 mg / ml, 25 mg / ml, 30 mg / ml, 35 mg / ml, 40 mg / ml, 45 mg / ml, or 50 mg / ml of an immunotherapeutic agent, e.g., an immune checkpoint inhibitor antibody or its antigen-binding fragment, and 20 mM sodium acetate, 0.2 mg / ml polysorbate 80, and 140 mM sodium chloride at pH 5.5. Additionally, the solution containing the checkpoint inhibitor antibody or its antigen-binding fragment may include, among many other compounds, histidine, mannitol, sucrose, trehalose, glycine, poly(ethylene) glycol, EDTA, methionine, any combination thereof, and many other compounds known in the relevant art.

[0882] In one embodiment, the pharmaceutical composition of the present disclosure comprises the following components: 5 mg to 500 mg of the immunotherapeutic agent of the present disclosure or its antigen-binding fragment, 10 mM histidine, 5% sucrose, and 0.01% polysorbate 80 (pH 5.8), and a non-polymorphic form, crystalline form, or crystalline salt form of Compound 1. The composition may be provided as a lyophilized powder. When the powder is reconstituted to its whole volume, the composition retains the same formulation. Alternatively, the powder may be reconstituted to half volume, in which case the composition comprises 10 mg to 500 mg of the immunotherapeutic agent of the present disclosure or its antigen-binding fragment, 20 mM histidine, 10% sucrose, and 0.02% polysorbate 80 (pH 5.8).

[0883] In one embodiment, a portion of the dose is administered by an intravenous bolus, and the remainder is administered by an infusion of an immunotherapeutic formulation. For example, an intravenous injection of about 0.001 mg / kg to about 200 mg / kg, for example, about 0.001 mg / kg to about 100 mg / kg, or about 0.001 mg / kg to about 50 mg / kg, or about 0.001 mg / kg to about 10 mg / kg of an immunotherapeutic agent or its antigen-binding fragment may be given by a bolus, and the remainder of the antibody dose may be administered by intravenous injection. A predetermined dose of the immunotherapeutic agent or its antigen-binding fragment may be administered over a period of, for example, 1 hour to 2 hours to 5 hours.

[0884] In additional embodiments, a portion of the dose is administered by subcutaneous injection and / or infusion in the form of a bolus, and the remainder is administered by infusion of an immunotherapeutic formulation. In some exemplary doses, the immunotherapeutic formulation may be administered subcutaneously at a dose of the immunotherapeutic agent or its antigen-binding fragment in the range of intravenous injection of about 0.001 mg / kg to about 200 mg / kg, e.g., about 0.001 mg / kg to about 100 mg / kg, or about 0.001 mg / kg to about 50 mg / kg, or about 0.001 mg / kg to about 10 mg / kg. In some embodiments, the dose may be given in a bolus, and the remainder of the immunotherapeutic dose may be administered by subcutaneous or intravenous injection. A predetermined dose of the immunotherapeutic agent or its antigen-binding fragment may be administered over a period of, e.g., 1 hour to 2 hours to 5 hours.

[0885] In this specification, the formulation may also comprise one or more active compounds required for the specific indication being treated, preferably those having complementary activities that do not have antagonistic effects on one another. For example, it may be desirable to provide one or more immunotherapies having different specificities. Alternatively or additionally, the composition may comprise anti-inflammatory agents, chemotherapy agents, cytotoxic agents, cytokines, growth inhibitors, and / or small molecule antagonists. These molecules are present in a suitable combination in amounts effective for the intended purpose.

[0886] The formulation used for in vivo administration must be sterile or nearly sterile. This is easily achieved by filtration through a sterile filter membrane.

[0887] In various embodiments, exemplary formulations of pharmaceutical compositions described herein may be prepared using methods widely known in the field of pharmaceutical formulations. Generally, such preparation methods may include the step of associating an active ingredient with an excipient or one or more other auxiliary ingredients, and then, if preferred, packaging the product into desired single-dose or multi-dose units.

[0888] In some embodiments, a composition comprising a non-polymorphic form, a crystalline form, or a crystalline salt form of compound 1 may also be delivered in a vesicle, and an immunotherapeutic agent may be delivered in the same liposomal formulation or a separate formulation suitable for the liposomal formulation comprising a non-polymorphic form, a crystalline form, or a crystalline salt form of compound 1. In some exemplary examples, a liposome comprising one or more liposomal surface moieties, e.g., polyethylene glycol, an antibody targeting a target tumor surface antigen, receptor, growth factor, glycoprotein, glycolipid, or neoantigen, and a fragment of the antibody thereof, is selectively transported to a specific cell or organ, thereby enhancing targeted drug delivery.

[0889] In another embodiment, the non-polymorphic form, crystalline form, or crystalline salt form of compound 1 may be delivered to vesicles, particularly liposomes (see Langer, Science 249: 1527-1533 (1990); Treat et al., in LIPOSOMES IN THE THERAPY OF INFECTIOUS DISEASE AND CANCER, Lopez-Berestein and Fidler (eds.), Liss, NY, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; generally see the same literature).

[0890] In another embodiment, a composition comprising a non-polymorphic form, crystalline form, or crystalline salt form of compound 1, or a combination thereof, or a composition comprising an immunotherapeutic agent may be delivered to a controlled release system. In one embodiment, a pump may be used (see references [quoted above: Langer; Sefton, CRC Crit. Ref. Biomed. Eng. 14: 201 (1987); Buchwald et al., Surgery 88: 507 (1980); Saudek et al., N. Engl. J. Med. 321: 574 (1989)]). In another embodiment, the controlled release of the non-polymorphic form, crystalline form, or crystalline salt form of compound 1 may comprise a polymeric material to provide sustained, intermediate, pulsating, or alternating release (see also [MEDICAL APPLICATIONS OF CONTROLLED RELEASE, Langer and Wise (eds.), CRC Pres., Boca Raton, Fla. (1974); CONTROLLED DRUG BIOAVAILABILITY, DRUG PRODUCT DESIGN AND PERFORMANCE, Smolen and Ball (eds.), Wiley, New York (1984); Ranger and Peppas, J. Macromol. Sci. Rev. Macromol. Chem. 23: 61 (1983)]; also [Levy et al., Science 228: 190 (1985); During et al., Ann. Neurol. 25: 351 (1989); Howard et al., J. See Neurosurg. 71: 105 (1989)]. Other controlled emission systems discussed in this review [Science 249: 1527-1533 (1990)] may be used.

[0891] The optimal concentration of the active ingredient(s) in the selected medium can be determined empirically according to procedures well known to those skilled in the art and will vary depending on the ultimate desired pharmaceutical formulation and intended use.

[0892] The present disclosure also provides a pharmaceutical pack or kit comprising one or more containers filled with one or more of the components of the pharmaceutical composition of the present disclosure, which will comprise at least a non-polymorphic form, a crystalline form, or a crystalline salt form of Compound 1 as described herein and one or more checkpoint inhibitor antibodies or antigen-binding fragments thereof. In other embodiments, the kit may comprise one or more additional containers providing pharmaceutically acceptable excipients, e.g., diluents. In one embodiment, the kit may comprise at least one container, wherein the container may comprise a non-polymorphic form, a crystalline form, or a crystalline salt form of Compound 1 of the present disclosure, a checkpoint inhibitor antibody or an antigen-binding fragment thereof. The kit may also comprise a set of instructions for preparing the final pharmaceutical composition and administering it to a subject requiring it for the treatment of a checkpoint molecule-mediated disease or disorder.

[0893] Labeled Compounds and Detection Methods

[0894] Another aspect relates to the labeled non-polymorphic, crystalline, or crystalline salt forms (radio-labeled, fluorescent-labeled, etc.) of the present invention, which are useful not only in imaging techniques but also in in vitro and in vivo assays for identifying and quantifying TAM kinases in tissue samples, including humans, and for identifying TAM kinase ligands by inhibitory binding of labeled compounds. Accordingly, the present invention includes a TAM kinase assay comprising such labeled compounds.

[0895] The present invention further comprises the isotope-labeled non-polymorphic form, crystalline form, or crystalline salt form of the present invention. The “isotope” or “radio-labeled” compound is the crystalline form or crystalline salt form of the present invention in which one or more atoms are replaced or substituted with atoms having an atomic mass or mass number different from the atomic mass or mass number typically found in nature (i.e., naturally occurring). Suitable radionuclides that may be incorporated into the non-polymorphic form, crystalline form, or crystalline salt form of the present invention are 2 H (sometimes denoted as D in the case of deuterium), 3 H (sometimes denoted as T in the case of tritium), 11 C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 18 F, 35 S, 36 Cl, 82 Br, 75 Br, 76 Br, 77 Br, 123 I, 124 I, 125 I and 131 It includes, but is not limited to, I. The radionuclides incorporated into the radiolabeled compounds of the present invention will vary depending on the specific application of the radiolabeled compounds. For example, in the case of in vitro metalloproteinase labeling and competitive testing, 3 H, 14 C, 82 Br, 125 I, 131 I or 35 Compounds containing S will generally be the most useful. For radiation imaging applications, 11 C, 18 F, 125 I, 123 I, 124 I, 131 I, 75 Br, 76 Br or77 Br will generally be the most useful. In some embodiments, the non-polymorphic form, crystalline form, or crystalline salt form described herein is replaced with deuterium, such as hydrogen bonded to a carbon atom. Since these compounds exhibit increased resistance to metabolism, they are useful for increasing the half-life of any compound when administered to mammals, particularly humans.

[0896] "Radiation-labeled" or "labeled compound" is understood to be a compound incorporating at least one radionuclide. In some embodiments, the radionuclide is 3 H, 14 C, 125 I, 35 S and 82 It is selected from the group consisting of Br.

[0897] The present invention may further include a synthesis method for incorporating a radioisotope into the non-polymorphic form, crystalline form, or crystalline salt form of the present invention. Synthesization methods for incorporating a radioisotope into an organic compound are well known in the art, and those skilled in the art will readily recognize a method applicable to the compound of the present invention.

[0898] The labeled compounds of the present invention can be used in screening assays to identify or evaluate compounds. For example, a newly synthesized or identified labeled compound (i.e., a test compound) can be evaluated for its ability to bind to TAM by monitoring changes in concentration upon contact with TAM kinase through tracking of the label. For example, the test compound (labeled) can be evaluated for its ability to reduce the binding of another compound (i.e., a standard compound) known to bind to TAM kinase. Thus, the ability of the test compound to compete with the standard compound for binding to TAM kinase is directly correlated with binding affinity. Conversely, in some other screening assays, the standard compound is labeled, while the test compound is not. Therefore, the relative binding affinity of the test compound is identified as the concentration of the labeled standard compound is monitored to evaluate the competition between the standard compound and the test compound.

[0899] Manufacturing and Examples

[0900] General experimental techniques

[0901] Water-based slurry experiment : The salt of compound 1, determined to have a water solubility of less than 1 mg / mL, was slurried in 20 mL of water at ambient temperature for 1 day. Then, the solid was collected by vacuum filtration and analyzed by XRPD.

[0902] Crash Cooling (CC) Concentrated solutions of Compound 1 and various counterions were prepared in MeOH at elevated temperatures while stirring. The lidded vials containing the hot solutions were transferred to a freezer (approx. -20°C) and rapidly cooled. The formed solids were collected. If no solids were present, additional crystallization techniques were used.

[0903] Crash Precipitation (CP)Clear solutions of compound 1 and a coformer were prepared in various solvents at room temperature. Aliquots of various inverse solvents were slowly added to the solution while gently stirring until a solid precipitated in the solution. The mixture was stirred for a certain period of time. The formed solid was collected by positive pressure filtration.

[0904] Fast Cooling (FC) Concentrated solutions of Compound 1 and various counterions were prepared in acetone or MeOH at elevated temperatures while stirring. The lidded vials containing the hot solutions were transferred to a benchtop at ambient temperature. The formed solids were collected. If no solids were present, additional crystallization techniques were used.

[0905] Fast Evaporation (FE) Clear solutions of Compound 1 and the co-form were prepared in various solvents. The vials were left uncovered, and the solvents were evaporated under ambient conditions.

[0906] Interconversion slurry A slurry of Compound 1 Type A was prepared by adding a sufficient amount of solid to a solvent system given under ambient conditions so that undissolved solid was present. Then, the mixture was stirred for an extended time to ensure saturation. Then, the solid of the form of interest was added to an aliquot of the saturated solution (filtered through a 0.2-µm nylon filter) so that undissolved solid was present. Then, the mixture was stirred for an extended time at ambient temperature, and the solid was isolated.

[0907] Simple interest technique Generally, isolation was performed rapidly after removing non-ambient samples from each temperature control device to minimize equilibrium with the ambient temperature prior to isolating the solid.

[0908] Liquid phase decanting: The suspension was centrifuged (if necessary), the liquid phase was discarded, and a portion of the solid isolated from the solution-based crystallization technique was collected by leaving the damped solid. Unless the solid is specified herein as "analyzed damp," the solid was briefly dried (e.g., air drying or drying under nitrogen).

[0909] Positive pressure filtration Solids were collected on a 0.2-µm nylon or PTFE filter by pressing the slurry through a syringe and a Swinnex filter holder assembly. Typically, air was blown over the filter with a 20-mL syringe to dry the solids briefly. Where "analyzed as moist" is indicated herein, the solids were kept moist in the mother liquor. Some samples were additionally dried briefly under a flow of nitrogen gas prior to analysis.

[0910] vacuum filtration The solid was collected on a paper or nylon filter by vacuum filtration and air-dried in the filter under reduced pressure for a short time before being transferred to a vial.

[0911] Reaction Crystallization (RC) A mixture of Compound 1 and various co-forming agents was prepared in an acetone slurry at elevated temperature such that the molar concentration of the co-forming agents was 2 times greater than API. The solution was stirred for a given time. Additional crystallization techniques were used when a clear solution was observed.

[0912] Stability test Salts of various compounds were placed in open vials within a 75% RH chamber (saturated sodium chloride solution). The RH chamber was placed in a 40°C oven for 15 to 16 days. At the end of the period, samples were analyzed by PLM and XRPD.

[0913] Slow Cooling (SC)Concentrated solutions of Compound 1 and various co-forms were prepared in various solvents while stirring at elevated temperatures. The vial lid was covered in the heated sample block, and the hot plate was turned off to allow the vial to cool slowly to ambient temperature in the heated vial block. Upon cooling to ambient temperature, the clear solution was further cooled in a refrigerator (5°C to 7°C) and / or a freezer (approx. -20°C). If no solid was present, additional crystallization techniques were used.

[0914] Slow evaporation Solutions were prepared in various solvents with stirring and, typically, filtered through a 0.2-µm nylon or PTFE filter. Each solution was allowed to evaporate from a covered vial (e.g., with a loosely closed lid or covered with perforated aluminum foil) under ambient conditions unless otherwise specified. Unless specified as partial evaporation (where solid remains with a small amount of solvent), the solution was allowed to evaporate dry, in which case the solid was isolated as described herein.

[0915] Solubility estimation : A fraction of various solvents was added to a measured amount of Compound 1 while stirring (typically sonication) at a specified temperature until complete dissolution was achieved as judged by visual observation. If dissolution occurred after adding the first fraction, the value was reported as ">". If dissolution did not occur, the value was reported as "<".

[0916] Estimation even if accepted : A fraction of water was added to the measured amount of various compound 1 salt while ultrasonically treating.

[0917] Slurry experiment Saturated solutions of Compound 1 and various co-forms were prepared in various solvents and solvent mixtures. The mixtures were stirred at ambient temperature and at elevated temperatures for the specified times. Solids were collected by the specified techniques, and additional crystallization techniques were used where appropriate.

[0918] Vacuum oven desolvation Desolvation was attempted on the salt of Compound 1, which was determined to be a solvate by various analytical methods. The samples were placed in a vacuum oven at a temperature ranging from ambient temperature to 80°C for a given time. The samples were analyzed by XRPD and / or TGA to determine the success of desolvation.

[0919] Vapor diffusion Concentrated solutions were prepared in various solvents and typically filtered through a 0.2-µm nylon or PTFE filter. The filtered solutions were dispensed into small vials and then placed into a larger vial containing the reverse solvent. The small vials were left uncovered, while the larger vial was covered to allow vapor diffusion. Any solids present were isolated as described herein.

[0920] steam stress The selected solid was transferred to a small vial and then placed inside a larger vial containing solvent. The small vial was left uncovered, while the larger vial was covered to allow vapor stress to develop at the specified temperature.

[0921] A co-former refers to one or more pharmaceutically acceptable bases and / or pharmaceutically acceptable acids disclosed herein associated with Compound 1. Exemplary co-formers used herein include fumaric acid, HCl, and phosphoric acid.

[0922] Device techniques

[0923] Differential Scanning Calorimetry (DSC)DSC was performed using a Mettler-Toledo DSC3+ differential scanning calorimeter. Temperature calibration was performed using adamantane, phenyl salicylate, indium, tin, and zinc. Samples were placed in completely sealed or open aluminum DSC pans, and their weights were accurately recorded. The weighed aluminum pans, which served as sample pans, were placed on the reference plane of the cell. Samples were analyzed from -30°C to 250°C at a ramp rate of 10°C / min. Thermograms were plotted against the reference temperature (x-axis), but results were reported against the sample temperature.

[0924] Dynamic Vapor Sorption (DVS)

[0925] a. VTI Automatic vapor sorption (VS) data were collected using a VTI SGA-100 vapor sorption analyzer. NaCl and PVP were used as calibration standards. Samples were dried prior to analysis. Adsorption and desorption data were collected under nitrogen purging in 10% RH increments over a range of 5% to 95% RH. The equilibrium criteria used for analysis were a maximum equilibrium time of 3 hours, with a weight change of less than 0.0100% within 5 minutes. Data regarding the initial moisture content of the samples were not corrected.

[0926] b. Intrinsic: Automatic vapor sorption (VS) data were collected using an Intrinsic DVS surface measurement system. Samples were not dried prior to analysis. Adsorption and desorption data were collected under nitrogen purging in 10% RH increments over a range of 5% to 95% RH. The equilibrium criteria used for analysis were a maximum equilibrium time of 3 hours, with a weight change of less than 0.0100% within 5 minutes. Data regarding the initial moisture content of the samples were not modified.

[0927] Hot stage Microscopy (HSM)Hot stage microscopy was performed using a Linkam hot stage (FTIR 600) mounted on a Leica DM LP microscope equipped with a SPOT Insight™ color digital camera. Temperature calibration was performed using the USP melting point standard. The sample was placed on a cover glass, and a second cover glass was placed over the sample. As the stage heated, each sample was observed visually using a 20x objective lens equipped with a crossed polarizer and a first-order red compensator. Images were captured using SPOT software (v. 4.5.9).

[0928] Optical microscope The sample was observed under a Motic or Wolfe optical microscope with a cross-polarizer or a Leica stereomicroscope with a primary red correction plate and a cross-polarizer.

[0929] pKa and log P determination : The determination of pKa and logP was performed by Pion Inc. / Sirius Analytical Instruments Ltd. in East Sussex, UK.

[0930] Solution proton nuclear magnetic resonance spectroscopy ( 1 HNMR) : Solution 1 1H NMR spectra were obtained from Spectral Data Services in Champaign, Illinois. Samples were prepared by dissolving approximately 5 mg to 10 mg of the sample in DMSO-d6.

[0931] Thermogravimetric Analysis (TGA)Thermogravimetric analysis was performed using a Mettler Toledo TGA / DSC3+ analyzer. Temperature calibration was performed using phenyl salicylate, indium, tin, and zinc. Samples were placed in aluminum pans. The open pans were inserted into a TG furnace. The furnace was heated under nitrogen. Each sample was heated from ambient temperature to 350°C at ramp rates of 2°C / min, 5°C / min, or 10°C / min. Thermograms were plotted against the reference temperature (x-axis), but results were reported based on the sample temperature.

[0932] X-ray powder diffraction (XRPD)

[0933] a. reflection XRPD patterns were collected using a PANalytical X'Pert PRO MPD diffractometer at room temperature (298 Kelvin) using an incident beam of Cu Kα radiation generated with a long microfocus source and a nickel filter. The diffractometer was constructed using symmetric Bragg-Brentano geometry. Prior to analysis, a silicon specimen (NIST SRM 640e) was analyzed to verify that the observed position of the Si 111 peak corresponded to the NIST-certified position. The specimen was packed into a well. An anticatter slit (SS) was used to minimize background generated by air. Soller slits for the incident and diffracted beams were used to minimize expansion from axial divergence. The diffraction patterns were collected using a scanning position-sensitive detector (X'Celerator) located 240 mm from the sample and data collector software v. It was collected using 2.2b.

[0934] b. Transmission:XRPD patterns were collected using a PANalytical X'Pert PRO MPD diffractometer with an incident beam of Cu radiation generated using an Optix long microfocus source at room temperature (298 Kelvin). Cu Kα X-rays were focused onto the detector through the specimen using an elliptical gradient multilayer mirror. Prior to analysis, a silicon specimen (NIST SRM 640e) was analyzed to verify that the observed position of the Si 111 peak corresponded to the NIST-certified position. The specimen was sandwiched between 3 µm-thick films and analyzed using transmission geometry. Background generated by air was minimized using beam-stopping, short anti-scattering extensions, and anti-scattering knife edges. Solar slits were used for both the incident and diffracted beams to minimize expansion from axial divergence. Diffraction patterns were collected using a scanning position-sensitive detector (X'Celerator) located 240 mm from the specimen and data collector software v. 2.2b.

[0935] XRPD Indexing

[0936] Indexing and structural improvement are computer-aided studies. Within the referenced figure for a given indexed XRPD pattern, the alignment between the allowed peak positions (indicated by bars) and the observed peaks indicates a consistent unit cell crystal. Successful indexing of the pattern indicates that the sample consists primarily of a single crystalline phase. Spatial groups corresponding to the assigned quenching symbols, unit cell parameters, and derived quantities are tabulated.

[0937] PD-1 antibody

[0938] The PD-1 antibody used in the example was purchased from BioXcell as cat#BE0146, clone RPMI-14, lot 780120J3.

[0939] Examples

[0940] Preparation Example 1: Synthesis of Compound 1

[0941] Step 1: N-(4-fluorophenyl)-N-(4-hydroxyphenyl)cyclopropane-1,1-dicaboxamide(4):

[0942]

[0943] 3-(ethyliminomethyleneamino)-N,N-dimethyl-propane-1-amine hydrochloride (EDCl) (10.31 g, 53.8 ml, 1.2 equivalents) was added to a solution of compound 2 (10 g, 44.80 ml, 1 equivalent) and compound 3 (5.87 g, 53.8 ml, 1.2 equivalents) in dimethylacetamide (DMA) (60 ml). The mixture was vigorously stirred at 20°C until the reaction was complete. The mixture was poured into aqueous (aq) saturated NaHCO3 (400 ml) and extracted with EtOAc (4 × 100 ml). The combined organic phase was washed with aqueous saturated NaCl (100 ml), dried with anhydrous Na2SO4, and concentrated. Compound 4 (21g, unrefined) (50% purity) was obtained. 1 ¹H NMR (400 MHz, DMSO- d 6 ) δ 10.16 (br s, 1H), 9.72 (br s, 1H), 7.61 (dd, 2H), 7.34 (d, 2H), 7.13 (t, 2H) 6.68 (d, 2H), 1.42 (s, 4H); C 17 H 15 MS (EI) for FN2O3, confirmed value 314.9 (MH+).

[0944] Step 2: Methyl 4-[4-[[1-[(4-fluorophenyl)carbamoyl]cyclopropane-carbonyl]amino]phenoxy]-7-methoxyquinoline-6-carbosylate(6):

[0945]

[0946] Compound in anisole (50 ml) 4(5.99g, 9.5mmol, 1.2 equivalents), compound 5(2g, 8.0mmol, 1.0 equivalent), Pd(OAc)2(89mg, 397.4μmol, 0.05 equivalents), rac -2-(Die- tertA mixture of -butylphosphino)-1,1'-binaphyl (TrixiePhos, 316.71 mg, 794.7 μmol, 0.1 equivalents) and K3PO4 (2.53 g, 11.9 mmol, 1.5 equivalents) was stirred at 110°C for 2 hours (h) under a nitrogen atmosphere. The mixture was filtered, and the filtrate was concentrated. The residue was purified by flash silica gel chromatography (1:1 petroleum ether:EtOAc to 20:1 EtOAc:MeOH). Compound 6 was obtained (2.6g, 61.8% yield). 1 H NMR (400 MHz, CDCl3) δ 9.38 (s, 1H), 8.80 (s, 1H), 8.63 (d, 2H), 7.64 (d, 2H), 7.54-7.41 (m, 3H), 7.18 (d, 2H), 7.09-7.01 (m, 2H), 6.43 (d, 1H), 4.05 (s, 3H), 3.97 (s, 3H), 1.78-1.72 (m, 2H), 1.69-1.63 (m, 2H); C 29 H 24 MS (EI) for FN3O6, confirmed value 530.0 (MH+).

[0947] Step 3: 4-[4-[[1-[(4-fluorophenyl)carbamoyl]cyclopropane-carbonyl]amino]phenoxy]-7-methoxyquinoline-6-carboxylic acid(7)

[0948]

[0949] Compound in tetrahydrofuran (THF) (15 ml) and MeOH (15 ml) 6 2M aqueous NaOH (7 mL, 4.1 equivalents) was added to a solution of (1.8 g, 3.4 mmol, 1 equivalent). The mixture was stirred at 6°C to 13°C for 4 hours. The pH of the mixture was adjusted to approximately 8 with 1M aqueous HCl, and the solvent was removed by concentration. Water (50 mL) was added, and the pH of the 1M aqueous HCl mixture was adjusted to approximately 6. The resulting precipitate was filtered, washed with water (2 × 10 mL), and dried under vacuum. Compound 7 was obtained (1.7g, 97.0% yield). 1 ¹H NMR (400 MHz, DMSO-d 6 ) δ 10.22 (s, 1H), 10.08 (s, 1H), 8.65 (d, 1H), 8.48 (s, 1H), 7.77 (d, 2H), 7.64 (dd, 2H) 7.47 (s, 1H), 7.25 (d, 2H), 7.15 (t, 2H), 6.45 (d, 1H), 3.96 (s, 3H), 1.47 (s, 4H); C 28 H 22 MS (EI) for FN3O6, confirmed value 516.1 (MH+).

[0950] Step 4: 1-N'-(4-fluorophenyl)-1-N-[4-[7-methoxy-6-(methylcarbamoyl)quinoline-4-yl]oxyphenyl]cyclopropane-1,1-dicaboxamide(1)

[0951]

[0952] Compound in DMF (10 ml) 7 Solutions of (300 mg, 582.0 μmol, 1 equivalent), HATU (332 mg, 873.2 μmol, 1.5 equivalents), and DIEA (301 mg, 2.3 mmol, 406 μl, 4 equivalents) were stirred at 6°C to 10°C for 1 hour. Methanolamine hydrochloride (79 mg, 1.2 mmol, 2.0 equivalents) was added, and the mixture was stirred at 6°C to 10°C for 17 hours. The mixture was filtered, and the resulting filtrate was purified by preparative HPLC (Column: Waters Xbridge 150 mm × 25 mm × 5 µm, Gradient: 33% to 63% acetonitrile in 10 mM aqueous NH4HCO3, Flow rate: 25 mL / min). Compound 1 was obtained (105.4 mg, 34.3% yield). 1 ¹H NMR (400 MHz, DMSO- d 6) δ 10.20 (s, 1H), 10.06 (s, 1H), 8.65 (d, 1H), 8.61 (s, 1H), 8.42-8.33 (m, 1H), 7.77 (d, 2H), 7.68-7.61 (m, 2H), 7.51 (s, 1H), 7.25 (d, 2H), 7.19-7.11 (m, 2H), 6.46 (d, 1H), 4.02 (s, 3H), 2.84 (d, 3H) 1.47 (s, 4H); C 29 H 25 MS (EI) for FN4O5, confirmed value 529.1 (MH+).

[0953] Example 1: Preparation of Compound 1 Fumara Form A

[0954] Fumaric acid (1 equivalent) in acetone was added to the free base (1 equivalent) of compound 1, and the resulting red slurry was stirred at about 50°C for 4 days. Then, the slurry was cooled at RT and stirred for an additional 1 day to obtain a pink slurry. Then, the solid was removed by positive pressure filtration to obtain a mixture of fumarate form A and free base form A.

[0955] Example 2: Preparation of Compound 1 Hemifumarate Form B

[0956] Fumaric acid (2 equivalents) in acetone was added to the free base (1 equivalent) of compound 1, and the resulting red slurry was stirred at about 50°C for 6 days to obtain a grayish-white slurry. Then, the solid was removed by positive pressure filtration of the hot solution to obtain hemifumarate form B.

[0957] Example 3: Preparation of Compound 1 HCl Form A

[0958] One equivalent of HCl was added to the free base of compound 1 in THF, and the resulting dark red slurry was stirred at RT for 3 days to obtain a thick grayish-white slurry. Then, the solid was removed by positive pressure filtration to obtain HCl form A.

[0959] Example 4: Preparation of Compound 1 HCl Form B

[0960] One equivalent of HCl was added to the free base of compound 1 in chloroform, and the resulting red slurry was stirred at about 50°C for 3 days to obtain a light pink slurry. Then, the solid was removed by positive pressure filtration to obtain HCl form B.

[0961] Example 5: Preparation of Compound 1 in HCl Form C

[0962] One equivalent of HCl was added to the free base of Compound 1 in methanol at a temperature of about 60°C to obtain a yellowish slurry. Then, the solution was cooled to about -20°C and kept cold for about 2 days to obtain a clear orange solution. Partial FE was obtained as a clear red solution, and then 4 volumes of reverse solvent MTBE were added, and the solution was stirred at RT for 1 day to obtain a grayish-white solid Compound 1 HCl form C separated by positive pressure filtration.

[0963] Example 6: Preparation of Compound 1 HCl Form D

[0964] Two equivalents of HCl were added to the free base of Compound 1 at approximately 50°C, and the resulting pink slurry was stirred at 50°C for 5 days. Solid Compound 1 HCl form D was separated by positive pressure filtration.

[0965] Example 7: Preparation of Compound 1 Form A

[0966] Compound 1 form A is likely the crystalline form of the free base of Compound 1, which is the most thermodynamically stable. Therefore, this form is formed through various procedures. A list of several possible procedures for obtaining Compound 1 form A is listed in the table below. This list is not exclusive, and there may actually be more procedures to produce this form.

[0967]

[0968] Example 8: Preparation of Compound 1 Form B

[0969] Compound 1 was dissolved in AcOH and crystallized by VD using diethyl ether as the inverse solvent.

[0970] Example 9: Preparation of Compound 1 Form C

[0971] Compound 1 was dissolved in HFIPA and crystallized by CP using MTBE as the inverse solvent.

[0972] Example 10: Preparation of Compound 1 Form D

[0973] Compound 1 was dissolved in methanol and crystallized by CC. Then, the mixture was slurried at 2°C to 8°C to obtain Form D.

[0974] Example 11: Preparation of Compound 1 Form E

[0975] Method A: Compound 1 was dissolved in THF and crystallized by CC.

[0976] Method B: Compound 1 was dissolved in 90:10 THF:water and precipitated by CP.

[0977] Example 12: Preparation of Compound 1 Form F

[0978] Method A: Compound 1 was dissolved in chloroform and crystallized by SE.

[0979] Method B: Compound 1 was slurried in chloroform.

[0980] Example 13: Preparation of Compound 1 Form G

[0981] Compound 1 was dissolved in chloroform, and the mixture was placed in a freezer to crystallize.

[0982] Example 14: Preparation of Compound 1 Form H

[0983] Form H was obtained by the amorphous compound 1 and the VS of DCM.

[0984] Example 15: Preparation of Compound 1 Form K

[0985] Compound 1 form K was prepared by desolvation of form F or form G, which are chloroform solvates.

[0986] Example 16: Preparation of Compound 1 Form O

[0987] Compound 1 form O was discovered while attempting to salt with various counterions in TFE-containing solvent systems, and it is likely a TFE solvate.

[0988] Example 17: Preparation of Compound 1 Phosphate Form A

[0989] 1 molar equivalent of phosphoric acid was added to a slurry of compound 1 in chloroform, and the resulting mixture was slurried at about 50°C for 3 days. The product was isolated by positive pressure filtration.

[0990] Example 18: Preparation of Compound 1 Form I

[0991] Compound 1 in a 90:10 THF / water mixture was rapidly precipitated with heptane and then stirred at freezing temperatures for 7 days.

[0992] Example 19: Preparation of Compound 1 Form J

[0993] Compound 1 was slurried in acetone for 14 days.

[0994] Example 20: Preparation of Compound 1 Form L

[0995] Compound 1 was slurried in chloroform for 14 days.

[0996] Example 21: Preparation of Compound 1 Form M

[0997] Compound 1 form E was dehydrated in a vacuum oven at about 77°C for 1 day.

[0998] Example 22: Preparation of Compound 1 Form N

[0999] Compound 1 was slurried in a 70:30 mixture of TFE / MTBE at room temperature for 7 days.

[1000] Example 23: In vivo study of the effect of Compound 1 on tumor angiogenesis

[1001] MC38 tumor-carrying animals were treated with a range of doses of Compound 1 (PO, qd; 3 mg / kg, 10 mg / kg, 30 mg / kg Compound 1) for 5 days. Tumors were analyzed for the presence of tumor microvessels by CD31 staining. Figures 1A and 1B compare tumor microvessels after administration of Compound 1 and the vehicle at different doses. The results show that Compound 1 inhibits in vivo angiogenesis. After treatment with 30 mg / kg Compound 1, the average number of vessels was significantly reduced compared to the vehicle (26.9 v. 17.6). A dose-dependent decrease in the presence of tumor microvessels was also observed.

[1002] Example 24: Effects of Compound 1 Combination Therapy on Immune Cells

[1003] MC38 tumor-bearing animals were treated with Compound 1 (PO, qd; 10 mg / kg) and anti-PD-1 antibody (IP, days 1, 2, 4, and 6; 5 mg / kg) for 7 days, and the tumors were analyzed for the presence of cytotoxic T-cells by CD8 staining. The results showed that the combination of Compound 1 and PD-1 increased natural killer (NK) cells and NK-T cells within the tumors. An increase in T cell and B cell levels was observed in the blood. Total macrophages and dendritic cells decreased, while g / mMDSC and M2 macrophages increased. In the blood, g / mMDSC macrophages and dendritic cells decreased, but an increase in M2 macrophages was also observed.

[1004] Figures 2A and 2B compare the number of CD8+ cells after treatment with PD-1, compound 1 + PD-1, and vehicle. After treatment with 10 mg / kg compound 1 + PD-1 (96.1 per 0.5 mm²), the average number of CD8+ T-cells increased significantly compared to treatment with vehicle (34.8 per 0.5 mm²) and PD-1 (49.8 per 0.5 mm²).

[1005] Example 25: Effect of Compound 1 combination therapy on tumor growth in MC38 model

[1006] MC38 tumor-bearing animals were treated with Compound 1 (3 mg / kg) + PD-1 (5 mg / kg), Compound 1 (3 mg / kg) + PD-L1 (5 mg / kg), and Compound 1 (3 mg / kg) + CTLA-4 (5 mg / kg), and tumor volume was analyzed. Figures 3A to 3C show tumor volume after treatment with the combination therapy. Compared to Compound 1, PD-1, PD-L1, and CTLA-4, the combination therapy significantly reduced tumor volume and significantly slowed or stopped tumor growth.

[1007] Example 26: In vivo evaluation of the efficacy of Compound 1 after oral administration to female Balb / c mice carrying CT26 tumors

[1008] During cancer progression, the expression of TAM (TYRO3, AXL, MER) receptor tyrosine kinases (RTKs) on various cell types influences a wide range of exogenous cellular characteristics in the tumor microenvironment (TME). Activation of these receptors on tumor cells increases tumor growth, survival, and metastatic potential, whereas activation on immune cell subtypes can lead to immunosuppression and resistance to chemotherapy.

[1009] Compound 1 was shown to be potent in vitro against MET, VEGFR2 and TAM RTK receptors, AXL, and MER. In addition to its effects on tumor cells, Compound 1 can also influence TAM RTK signaling in tumor-associated macrophages, which inhibits apoptotic cell elimination and polarizes macrophages into an immune-accepting M1 phenotype. Treatment with Compound 1 also resulted in reduced angiogenesis and a decrease in angiogenic capacity in TME, as evidenced by VEGFR2 inhibition. Collectively, these effects were shown to lead to significant improvements in tumor growth inhibition (TGI) when combined with an anti-PD-1 antibody.

[1010] In this study, the inhibitory effects of Compound 1 as a monoagonist and in combination with an anti-PD-1 inhibitor were investigated in Balb / c mice transplanted with CT26 colorectal carcinoma cells. CT26 cells inoculated into these mice are highly oncogenic (Brattain et al. 1980) and share molecular characteristics with undifferentiated, invasive human colorectal carcinoma cells (Castle et al. 2014). Therefore, the CT26 cell line can serve as a valid cell model for metastatic and poorly differentiated human colorectal cancer. Furthermore, this model has been reported to express significant levels of RTKs, indicating a potential dependence on these signaling pathways for the maintenance of the oncogenic phenotype in vivo (Poryzybyszewska et al. 2017).

[1011] method

[1012] Cell line culture and maintenance

[1013] CT26 colon carcinoma cells (ATCC, CRL-2638) from the Exelixis pharmacology cell bank were thawed in a T-75 flask (Corning, 43064U) using fresh culture medium, namely RPMI-1640 containing 10% fetal bovine serum (GIBCO, A384002). The cells were incubated at 37°C in an incubator humidified with 5% CO2 and grown to 80% to 90% confluence by the time of transplantation.

[1014] transplantation

[1015] Mice were anesthetized with isoflurane during transplantation. One million CT26 cells in 0.1 ml of serum-free culture medium containing equal volumes of Matrigel (Corning, 354235, protein concentration 11.0 mg / ml, endotoxin level < 1.5 unit / ml) were transplanted subcutaneously (sc) into the right hind flank of 160 Balb / C mice using a 25G needle attached to a 1 ml syringe. The cells were prepared in 50 ml tubes, kept on ice, and mixed before loading into each syringe.

[1016] Random assignment

[1017] Twelve mice were selected after randomization using Studylog software to match an average size of 180 mm³. The mice were separated into a treatment group of 5 mice per cage; 2 cages per group.

[1018] Administration Management

[1019] Oral doses were administered via a -20ga, 1.5" stainless steel needle with a silicone tip (VWR, 20068-666). Mice were administered daily according to body weight. Survival was monitored for up to 40 days. Compound 1 (EXEL-04621820), Lot 11 from the Exelisys Compound Repository was used as a monoagonist or in combination with anti-PD-1 (BioExcel cat#BE0146, clone RPMI-14, Lot 780120J3).

[1020] observation

[1021] Tumor volume and body weight of all animals were measured twice a week using Studylog software.

[1022] Statistical methods

[1023] Significance values ​​represent differences compared to the vehicle or anti-PD-1 treatment groups and were determined using the non-parametric Mann-Whitney U-test. The described significance levels are as follows: * : p<.05, ** : p<.01, *** : p<.001, **** : p<.0001

[1024] result

[1025] Treatment of xenografted mice with Compound 1 at doses of 1 mg / kg / day, 3 mg / kg / day, 10 mg / kg / day, and 30 mg / kg / day for 40 days resulted in a delay in tumor growth. Further delay in tumor growth was observed when Compound 1 treatment was combined with 5 mg / kg anti-PD-1 (Fig. 4). Survival benefit and dose-response were observed in the combination of the Compound 1 monotherapy and PD-1. Statistical significance (P value < 0.0001) was observed between Compound 1 ...

Claims

Claim 1 A pharmaceutical composition for treating colorectal cancer in subjects, comprising: the following compound 1 in a dosage of 5 mg to 100 mg: A pharmaceutical composition comprising a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient, administered in combination with a therapeutically effective amount of atezolizumab. Claim 2 A pharmaceutical composition according to claim 1, wherein compound 1 or a pharmaceutically acceptable salt thereof is administered orally once a day (qd) or twice a day (bid). Claim 3 A pharmaceutical composition according to paragraph 2, wherein the dosage of compound 1 or a pharmaceutically acceptable salt thereof is 5 mg to 80 mg. Claim 4 A pharmaceutical composition according to any one of claims 1 to 3, wherein the dosage of compound 1 or a pharmaceutically acceptable salt thereof is selected from 10 mg, 20 mg, 40 mg, 60 mg, and 80 mg. Claim 5 A pharmaceutical composition according to any one of claims 1 to 3, wherein atezolizumab is administered intravenously (IV) to a subject. Claim 6 A pharmaceutical composition according to any one of claims 1 to 3, wherein atezolizumab is administered once every 2 weeks, once every 3 weeks, or once every 4 weeks during the treatment period. Claim 7 A pharmaceutical composition according to claim 6, wherein the dosage of atezolizumab is 800 mg to 1700 mg. Claim 8 A pharmaceutical composition according to any one of claims 1 to 3, wherein the dosage of atezolizumab is 840 mg administered once every 2 weeks, 1200 mg administered once every 3 weeks, or 1680 mg administered once every 4 weeks. Claim 9 In claim 8, the above atezolizumab is administered to a subject in the form of an IV unit dose, wherein the dose form comprises 840 mg, 1200 mg, or 1680 mg of atezolizumab, water, glacial acetic acid, L-histidine, polysorbate 20, and sucrose, a pharmaceutical composition. Claim 10 In any one of claims 1 to 3, the pharmaceutical composition administered as a compound 1 comprising: a. 25% to 35% by weight of compound 1 or a pharmaceutically acceptable salt thereof; b. 37% to 43% by weight of microcrystalline cellulose; c. 18% to 22% by weight of anhydrous lactose; d. 2% to 6% by weight of hydroxypropyl cellulose; e. 5% to 7% by weight of croscarmellose sodium; f. 0.2% to 0.4% by weight of colloidal silicon dioxide; and g. 0.5% to 3.5% by weight of magnesium stearate. Claim 11 In any one of claims 1 to 3, the pharmaceutical composition administered as a pharmaceutical composition comprising: a. 25% to 35% by weight of compound 1 or a pharmaceutically acceptable salt thereof; b. 35% to 40% by weight of microcrystalline cellulose; c. 16% to 22% by weight of anhydrous lactose; d. 3% to 7% by weight of hydroxypropyl cellulose; e. 3% to 7% by weight of croscarmellose sodium; f. 0.1% to 0.5% by weight of colloidal silicon dioxide; and g. 0.5% to 3.5% by weight of stearic acid. Claim 12 A pharmaceutical composition according to any one of claims 1 to 3, wherein the colorectal cancer is an inoperable, locally advanced, metastatic, or recurrent solid tumor. Claim 13 A pharmaceutical composition according to claim 12, wherein the solid tumor is unresectable or metastatic, and no life-prolonging therapy is available or available therapy is tolerable or no longer effective. Claim 14 A pharmaceutical composition according to claim 1, wherein the colorectal cancer is right colorectal cancer (RCRC) or left colorectal cancer (LCRC). Claim 15 In claim 1, the subject is a pharmaceutical composition that has received prior anticancer therapy. Claim 16 A pharmaceutical composition according to claim 15, wherein the aforementioned prior anticancer therapy comprises a fluoropyrimidine combined with oxaliplatin or irinotecan. Claim 17 delete Claim 18 delete Claim 19 delete Claim 20 delete Claim 21 delete Claim 22 delete Claim 23 delete Claim 24 delete Claim 25 delete Claim 26 delete Claim 27 delete Claim 28 delete Claim 29 delete Claim 30 delete Claim 31 delete Claim 32 delete Claim 33 delete Claim 34 delete Claim 35 delete Claim 36 delete Claim 37 delete Claim 38 delete Claim 39 delete Claim 40 delete Claim 41 delete Claim 42 delete Claim 43 delete Claim 44 delete Claim 45 delete Claim 46 delete Claim 47 delete Claim 48 delete Claim 49 delete Claim 50 delete Claim 51 delete Claim 52 delete Claim 53 delete Claim 54 delete Claim 55 delete Claim 56 delete Claim 57 delete Claim 58 delete Claim 59 delete Claim 60 delete Claim 61 delete Claim 62 delete Claim 63 delete Claim 64 delete

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