Metal salts and their use

Abstract

The present invention relates to metal salts of 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]methyl-(2,2-diphenylethyl)amino]butoxy]phenyl]acetic acid, which exhibit improved physical properties and stability. The present invention also relates to pharmaceutical compositions comprising an effective amount of the metal salts, as well as methods for treating cancer, comprising administering to a subject in need thereof a pharmaceutical composition comprising a salt of the invention.

Description

Background Art

[0001] Liver X receptor (LXR) is a nuclear receptor transcription factor. LXR modulators have been found to be useful in the treatment of various diseases including cancer. There is a need to provide salts of such compounds with improved stability and physical properties.

Summary of the Invention

Means for Solving the Problems

[0002] The present invention provides a metal salt of an LXRβ agonist. The present invention also provides such a metal salt of an LXRβ agonist, a method for producing a pharmaceutical composition containing the metal salt, and a method for treating cancer with such a composition.

[0003] Thus, in one aspect, the present invention features a metal salt (e.g., a pharmaceutically acceptable metal salt) of 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]methyl-(2,2-diphenylethyl)amino]butoxy]phenyl]acetic acid (Compound 1).

[0004] In some embodiments, the metal salt of 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]methyl-(2,2-diphenylethyl)amino]butoxy]phenyl]acetic acid is a polyvalent metal salt.

[0005] In some embodiments, the metal salt of 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]methyl-(2,2-diphenylethyl)amino]butoxy]phenyl]acetic acid is a zinc salt, e.g., a 2:1 (Compound 1:zinc) salt.

[0006] In some embodiments, the metal salt of 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]methyl-(2,2-diphenylethyl)amino]butoxy]phenyl]acetic acid is an aluminum salt, e.g., a 3:1 (Compound 1:aluminum) salt.

[0007] In some embodiments, the metal salt of 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]methyl-(2,2-diphenylethyl)amino]butoxy]phenyl]acetic acid is a bismuth salt, such as a 3:1 (compound 1:bismuth) salt.

[0008] In some embodiments, the metal salt of 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]methyl-(2,2-diphenylethyl)amino]butoxy]phenyl]acetic acid is a calcium salt, such as a 2:1 (compound 1:calcium) salt.

[0009] In some embodiments, the metal salt is amorphous. In some embodiments, the metal salt (e.g., amorphous zinc salt) has a peak with an increased intensity relative to the free acid at about 1590 ± 10 cm -1 and a peak with a decreased intensity relative to the free acid at about 1710 ± 10 cm -1 In some embodiments, the metal salt has a mass loss of less than 1% up to decomposition, as measured by thermogravimetric analysis.

[0010] In other aspects, the invention features a method for making a zinc salt of compound 1 (e.g., a 2:1 zinc salt). The method includes combining compound 1 or a salt thereof (e.g., a 1:1 sodium salt or the free compound) and a zinc salt (e.g., zinc chloride or zinc acetate) in an amount sufficient to form the zinc salt of compound 1.

[0011] In some embodiments of the method for generating a zinc salt, the method includes dissolving Compound 1 or a salt thereof and the zinc salt in a solvent to form a mixture. In some embodiments, the solvent is water. In some embodiments, the solvent is a mixture of an organic solvent (e.g., methanol) and water (e.g., a 9:1 volume ratio of organic solvent to water). In some embodiments of the method for generating a zinc salt, the method further includes cycling the temperature of the mixture between ambient temperature and 40°C. In some embodiments of the method for generating a zinc salt, the cycling is performed for 24 hours.

[0012] In other aspects, the present invention features a pharmaceutically acceptable zinc salt of Compound 1 produced by any of the methods described above.

[0013] In other aspects, the present invention features a method for producing an aluminum salt (e.g., a 3:1 aluminum salt) of Compound 1. The method includes combining Compound 1 or a salt thereof (e.g., a 1:1 sodium salt or the free compound) and an aluminum salt (e.g., aluminum sulfate) in an amount sufficient to produce the aluminum salt of Compound 1.

[0014] In other aspects, the present invention features a pharmaceutical composition containing any of the metal salts described above and a pharmaceutically acceptable excipient.

[0015] In some embodiments, the pharmaceutical composition contains less than 1.5% by weight of sodium. In some embodiments, the pharmaceutical composition substantially does not contain the 1:1 sodium salt of Compound 1. In some embodiments, the pharmaceutical composition is in unit dosage form.

[0016] In other aspects, the present invention features a method for treating cancer. The method includes administering any of the salts or pharmaceutical compositions described above in an effective amount.

[0017] In some embodiments of the method of treating cancer, the subject has cancer that did not respond to a previously administered immunotherapy (e.g., the subject's cancer has progressed despite treatment with immunotherapy).

[0018] In some embodiments of the method of treating cancer, the cancer is resistant to immunotherapy (e.g., the cancer has been determined to be resistant to immunotherapy based on genetic markers in a sample or levels of MDSCs (e.g., monocytic and / or granulocytic MDSCs), or is a cancer that did not respond to immunotherapy and is suspected of being resistant to immunotherapy).

[0019] In other aspects, the present invention features a method of treating cancer that did not respond to immunotherapy in a subject. The method includes administering to the subject, in combination with immunotherapy, an effective amount of any of the salts or pharmaceutical compositions described above.

[0020] In other aspects, the present invention features a method of treating cancer that is resistant to immunotherapy in a subject. The method includes administering to the subject, in combination with immunotherapy, an effective amount of any of the salts or pharmaceutical compositions described above.

[0021] In some embodiments of any of the methods of treating cancer, the cancer is breast cancer, colon cancer, renal cell carcinoma, lung cancer (e.g., non-small cell lung cancer), hepatocellular carcinoma, gastric cancer, ovarian cancer, pancreatic cancer, esophageal cancer, prostate cancer, sarcoma, glioblastoma, diffuse large B-cell lymphoma, leukemia, or melanoma. In some embodiments, the cancer is metastatic cancer. In some embodiments of any of the methods of treating cancer, the effective amount is an amount effective to inhibit metastatic colony formation of the cancer.

[0022] In certain embodiments of any of the methods of treating cancer, the cancer is drug-resistant cancer or did not respond to prior treatment (e.g., vemurafenib, dacarbazine, CTLA-4 inhibitor, PD-1 inhibitor, interferon therapy, BRAF inhibitor, MEK inhibitor, radiation therapy, temozolomide, irinotecan, CAR-T therapy, Herceptin, Perjeta, tamoxifen, Xeloda, docetaxel, platinum agents such as carboplatin, taxanes such as paclitaxel and docetaxel, ALK inhibitor, MET inhibitor, Alimta, Abraxane, Adriamycin, gemcitabine, Avastin, Halaven, neratinib, PARP inhibitor, brilanestrant, mTOR inhibitor, topotecan, Gemzar, VEGFR2 inhibitor, folate receptor antagonist, demcizumab, fosbretabulin or cancer resistant to PD-L1 inhibitor or cancer that did not respond to previous treatment with these).

[0023] In some embodiments of any method of treating cancer, the immunotherapy is, if present, a CTLA-4 inhibitor, a PD-1 inhibitor, a PD-L1 inhibitor, or adoptive T cell transfer therapy. In some embodiments, the immunotherapy comprises a PD-1 inhibitor, such as a PD-1 antibody, a PD-L1 inhibitor, such as a PD-L1 antibody, a CTLA-4 inhibitor, such as a CTLA-4 antibody, a CSF-1R inhibitor, an IDO inhibitor, an A1 adenosine inhibitor, an A2A adenosine inhibitor, an A2B adenosine inhibitor, an A3A adenosine inhibitor, an arginase inhibitor, or an HDAC inhibitor. In some embodiments, the immunotherapy comprises a PD-1 inhibitor (e.g., nivolumab, pembrolizumab, pidilizumab, BMS 936559, and atezolizumab). In some embodiments, the immunotherapy comprises a PD-L1 inhibitor (e.g., atezolizumab and durvalumab). In some embodiments, the immunotherapy comprises a CTLA-4 inhibitor (e.g., ipilimumab). In some embodiments, the immunotherapy comprises a CSF-1R inhibitor (e.g., pexidartinib and 4-(2,4-difluoroanilino)-7-ethoxy-6-(4-methylpiperazin-1-yl)quinoline-3-carboxamide). In some embodiments, the immunotherapy comprises an IDO inhibitor (e.g., norharman, rosmarinic acid, and α-methyl-tryptophan). In some embodiments, the immunotherapy comprises an A1 adenosine inhibitor (e.g., 8-cyclopentyl-1,3-dimethylxanthine, 8-cyclopentyl-1,3-dipropylxanthine, 8-phenyl-1,3-dipropylxanthine, bambifylline, BG-9719, tonapofylline, FK-453, FK-838, lofofylline, or N-0861). In some embodiments, the immunotherapy comprises an A2A adenosine inhibitor (e.g., ATL-4444, istradefylline, MSX-3, preladenant, SCH-58261, SCH-412348, SCH-442416, 2-butyl-9-methyl-8-(triazol-2-yl)purin-6-amine, VER-6623, VER-6947, VER-7835, viadenant, or Z / M-241,385).In some embodiments, the immunotherapy comprises an A2B adenosine inhibitor (e.g., N-[5-(1-cyclopropyl-2,6-dioxo-3-propyl-7H-purin-8-yl)pyridin-2-yl]-N-ethylpyridine-3-carboxamide, 3-ethyl-1-propyl-8-[1-[[3-(trifluoromethyl)phenyl]methyl]pyrazol-4-yl]-7H-purine-2,6-dione, MRS-1706, MRS-1754, N-[2-[[2-phenyl-6-[4-(3-phenylpropyl)piperazine-1-carbonyl]-7H-pyrrolo[2,3-d]pyrimidin-4-yl]amino]ethyl]acetamide, PSB-603, PSB-0788, or PSB-1115). In some embodiments, the immunotherapy comprises an A3A adenosine inhibitor (e.g., KF-26777, MRS-545, MRS-1191, MRS-1220, MRS-1334, propyl 6-ethyl-5-ethylsulfanylcarbonyl-2-phenyl-4-propylpyridine-3-carboxylate, MRS-3777, MRE-3005-F20, MRE-3008-F20, PSB-11, OT-7999, VUF-5574, and SSR161421). In some embodiments, the immunotherapy comprises an arginase inhibitor (e.g., an arginase antibody, (2s)-(+)-amino-5-iodoacetamidopentanoic acid, NG-hydroxy-L-arginine, (2S)-(+)-amino-6-iodoacetamidohexanoic acid, or (R)-2-amino-6-borono-2-(2-(piperidin-1-yl)ethyl)hexanoic acid). In some embodiments, the immunotherapy comprises an HDAC inhibitor (e.g., valproic acid, SAHA, or romidepsin).

[0024] In any other embodiment of the method of treating cancer, the method further comprises administering to the subject a further anti-cancer therapeutic agent (e.g., an anti-proliferative agent).

[0025] In certain embodiments, the anti-proliferative agent is a chemotherapeutic or cytotoxic agent, a differentiation inducer (e.g., retinoic acid, vitamin D, cytokine), a hormonal agent, an immunological agent or an anti-angiogenic agent. Chemotherapeutics and cytotoxic agents include, but are not limited to, alkylating agents, cytotoxic antibiotics, antimetabolites, vinca alkaloids, etoposide, etc. (e.g., paclitaxel, taxol, docetaxel, taxotere, cisplatin). A list of additional compounds having anti-proliferative activity can be found in L. Brunton, B. Chabner and B. Knollman (eds). Goodman and Gilman’s The Pharmacological Basis of Therapeutics, Twelfth Edition, 2011, McGraw Hill Companies, New York, NY.

[0026] In certain embodiments, the anti-proliferative agent is a PD-1 inhibitor, a VEGF inhibitor, a VEGFR2 inhibitor, a PD-L1 inhibitor, a BRAF inhibitor, a CTLA-4 inhibitor, a MEK inhibitor, an ERK inhibitor, vemurafenib, dacarbazine, trametinib, dabrafenib, durvalumab, an mTOR inhibitor, CAR-T therapy, abiraterone, enzalutamide, apalutamide, 5-fluorouracil (5-FU), FOLFOX (i.e., folinic acid, 5-fluorouracil, and oxaliplatin), FOLFIRI (i.e., folinic acid, 5-fluorouracil, and irinotecan), Herceptin, Xeloda, a PD-1 antibody (e.g., pembrolizumab or nivolumab), a PD-L1 antibody, a CTLA-4 antibody (e.g., ipilimumab), ramucirumab, linsitinib, gemcitabine, vedotin, ANG1005 and / or ANG4043.

[0027] In some embodiments of any of the methods of treating cancer, the cancer is renal cell carcinoma and the anti-proliferative agent is a PD-1 inhibitor, a PDL-1 inhibitor or an mTOR inhibitor. In other embodiments, the cancer is diffuse large B-cell lymphoma and the anti-proliferative agent is CAR-T therapy. In certain embodiments, the cancer is prostate cancer and the anti-proliferative agent is abiraterone, enzalutamide or apalutamide. In some embodiments, the cancer is hepatocellular carcinoma, gastric cancer or esophageal cancer and the anti-proliferative agent is 5-FU, FOLFOX, FOLFIRI, Herceptin or Xeloda. In some embodiments, the cancer is sarcoma and the anti-proliferative agent is gemcitabine. In other embodiments, the cancer is pancreatic cancer and the anti-proliferative agent is irinotecan, cisplatin, Abraxane, taxane (e.g., paclitaxel or docetaxel) or capecitabine.

[0028] The method of treating cancer may further comprise administering an alkylating agent, a platinum agent, an antimetabolite, a topoisomerase inhibitor, an antitumor antibiotic, a mitotic inhibitor, an aromatase inhibitor, a thymidylate synthase inhibitor, a DNA antagonist, a farnesyl transferase inhibitor, a pump inhibitor, a histone acetyl transferase inhibitor, a metalloproteinase inhibitor, a ribonucleoside reductase inhibitor, a TNF alpha agonist / antagonist, an endothelin A receptor antagonist, a retinoic acid receptor agonist, an immunomodulatory agent, a hormone and an antihormone agent, a photodynamic agent, a tyrosine kinase inhibitor, an antisense compound, a corticosteroid, an HSP90 inhibitor, a proteasome inhibitor (e.g., marizomib), a CD40 inhibitor, an anti-CSI antibody, an FGFR3 inhibitor, a VEGF inhibitor, a MEK inhibitor, a cyclin D1 inhibitor, an NF-kB inhibitor, an anthracycline, a histone deacetylase, a kinesin inhibitor, a phosphatase inhibitor, a COX2 inhibitor, an mTOR inhibitor, a calcineurin antagonist, an anti-proliferative agent such as an IMiD, and / or other agents used to treat proliferative diseases.

[0029] In some embodiments of any of the methods of treating cancer, the cancer is breast cancer, such as triple negative breast cancer, colon cancer, renal cell carcinoma, lung cancer (e.g., non-small cell lung cancer), hepatocellular carcinoma, gastric cancer, ovarian cancer, pancreatic cancer, esophageal cancer, prostate cancer, sarcoma, glioblastoma, diffuse large B-cell lymphoma, leukemia (e.g., acute myeloid leukemia) or melanoma. In some embodiments of any of the methods of treating cancer, the cancer is melanoma. In some embodiments of any of the methods of treating cancer, the cancer is breast cancer. In some embodiments of any of the methods of treating cancer, the cancer is renal cell carcinoma. In some embodiments of any of the methods of treating cancer, the cancer is pancreatic cancer. In some embodiments of any of the methods of treating cancer, the cancer is non-small cell lung cancer. In some embodiments of any of the methods of treating cancer, the cancer is colon cancer. In some embodiments of any of the methods of treating cancer, the cancer is ovarian cancer. In some embodiments of any of the methods of treating cancer, the cancer is glioblastoma. In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is diffuse large B-cell lymphoma. In some embodiments, the cancer is leukemia (e.g., acute myeloid leukemia).

[0030] In certain embodiments of any of the methods of treating cancer, the cancer is melanoma (e.g., metastatic melanoma) that is resistant to vemurafenib, dacarbazine, interferon therapy, CTLA-4 inhibitors, BRAF inhibitors, MEK inhibitors, PD1 inhibitors, PDL-1 inhibitors, and / or CAR-T therapy, or that did not respond to previous treatment with these. In some embodiments of any of the methods of treating cancer, the cancer is glioblastoma that is resistant to temozolomide, radiation therapy, Avastin, irinotecan, VEGFR2 inhibitors, CAR-T therapy, and / or mTOR inhibitors, or that did not respond to previous treatment with these. In some embodiments of any of the methods of treating cancer, the cancer is non-small cell lung cancer, such as metastatic non-small cell lung cancer (e.g., EGFR wild-type non-small cell lung cancer and / or squamous non-small cell lung cancer), etc., that is resistant to EGFR inhibitors, platinum agents (e.g., carboplatin), Avastin, ALK inhibitors, MET inhibitors, taxanes (e.g., paclitaxel and / or doceltaxel), gemzar, alimta, radiation therapy, PD-1 inhibitors, PD-L1 inhibitors, and / or CAR-T therapy, or that did not respond to previous treatment with these. In some embodiments of any of the methods of treating cancer, the cancer is breast cancer (e.g., triple-negative breast cancer) that is resistant to Herceptin, Perjeta, tamoxifen, Xeloda, docetaxel, carboplatin, paclitaxel, Abraxane, Adriamycin, gemcitabine, Avastin, Halaven, neratinib, PARP inhibitors, PD-1 inhibitors, PD-L1 inhibitors, CAR-T therapy, apalutamide, and / or mTOR inhibitors, or that did not respond to previous treatment with these. In some embodiments of any of the methods of treating cancer, the cancer is ovarian cancer (e.g., advanced ovarian cancer) that is resistant to PARP inhibitors, Avastin, platinum agents, such as carboplatin, etc., paclitaxel, docetaxel, topotecan, gemzar, VEGR2 inhibitors, folate receptor antagonists, PD-1 inhibitors, PD-L1 inhibitors, CAR-T therapy, demcizumab, and / or fosbretabulin, or that did not respond to previous treatment with these.

[0031] In some embodiments of any of the methods of treating cancer, if present, additional anti-cancer therapies include chemotherapy.

[0032] In some embodiments of any of the methods of treating cancer, chemotherapy includes docetaxel. In some embodiments, the method includes administering an effective amount of docetaxel to the subject once every 7 days. In some embodiments, the effective amount of docetaxel is at least 28 mg / m 2 . In some embodiments, the effective amount of docetaxel is about 28 to about 35 mg / m 2 .

[0033] In some embodiments of any of the methods of treating cancer, other anti-cancer therapies include chemotherapy and immunotherapy. In some embodiments of any of the methods of treating cancer, the anti-cancer therapy includes carboplatin or cisplatin, pemetrexed, and pembrolizumab. In some embodiments, the method includes administering an effective amount of pembrolizumab to the subject once every 21 days. In some embodiments, the effective amount of pembrolizumab is about 200 mg. In some embodiments, the method includes administering an effective amount of carboplatin or cisplatin to the subject once every 21 days. In some embodiments, the effective amount of carboplatin or cisplatin is calculated using the formula: total dose (mg) = (area under the target curve) × (glomerular filtration rate of the subject + 25), where the area under the target curve is 4 to 6 mg / mL·min and the glomerular filtration rate of the subject is measured by Cr-EDTA clearance. In some embodiments, the effective amount of carboplatin or cisplatin is about 300 to about 360 mg / m 2 . In some embodiments, the method includes administering an effective amount of pemetrexed to the subject once every 21 days. In some embodiments, the effective amount of pemetrexed is 500 mg / m 2It is. In some embodiments of any of the methods for treating cancer, the method further comprises administering to the subject an effective amount of folic acid, vitamin B12, and / or corticosteroid. In some embodiments, the method comprises administering to the subject an effective amount of corticosteroid twice a day for 3 days prior to the administration of pemetrexed.

[0034] In some embodiments of any of the methods for treating cancer, the method further comprises administering to the subject an effective amount of a statin (e.g., rosuvastatin or atorvastatin).

[0035] In some embodiments of any of the methods for treating cancer, the method further comprises administering to the subject an effective amount of an antiemetic (e.g., ondansetron, granisetron, palonosetron, metoclopramide, haloperidol, dexamethasone, aprepitant, fosaprepitant, lorazepam, dronabinol, prochlorperazine, or chlorpromazine), an antidiarrheal agent (e.g., an opioid agonist or octreotide), an appetite stimulant (e.g., megestrol acetate, metoclopramide, dronabinol, prednisone, or dexamethasone), a systemic stimulant, a bisphosphonate (e.g., etidronate, clodronate, tiludronate, pamidronate, neridronate, oplatronate, alendronate, ibandronate, risedronate, or zoledronate), a gonadotropin-releasing hormone agonist (e.g., buserelin, histrelin, leuprolide, triptorelin, goserelin, or nafarelin), and / or a growth factor (e.g., filgrastim).

[0036] In some embodiments of any of the methods of treating cancer, the cancer is resistant to anti-cancer therapies (e.g., platinum-containing chemotherapy, PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, mitotic inhibitors, topoisomerase inhibitors, antimetabolites, angiogenesis inhibitors, kinase inhibitors, and / or alkylating agents). In some embodiments of any of the methods of treating cancer, during or after treatment with an anti-cancer therapy (e.g., platinum-containing chemotherapy, PD-1 inhibitors, PD-L1 inhibitors, angiogenesis inhibitors, kinase inhibitors, and / or alkylating agents), the cancer progresses. In some embodiments of any of the methods of treating cancer, the cancer has been determined or is predicted to be resistant to an anti-cancer therapy (e.g., PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, topoisomerase inhibitors, antimetabolites, angiogenesis inhibitors, kinase inhibitors, and / or alkylating agents).

[0037] In some embodiments of any of the methods of treating cancer, the cancer has a PD-L1 expression level of less than 1% when tested by immunohistochemical assay (e.g., immunohistochemical assay using the proportion of PD-L1 positive cells (tumor proportion score) in tumor cells). In some embodiments of any of the methods of treating cancer, the cancer has a PD-L1 expression level of about 1% when tested by immunohistochemical assay (e.g., immunohistochemical assay using the proportion of PD-L1 positive cells in tumor cells). In some embodiments of any of the methods of treating cancer, the cancer has a PD-L1 expression level of about 1 to about 49% (e.g., about 1 to about 20%, about 5 to about 30%, about 15 to about 40%, about 25 to about 49%) when tested by immunohistochemical assay (e.g., immunohistochemical assay using the proportion of PD-L1 positive cells in tumor cells). In some embodiments of any of the methods of treating cancer, the cancer is metastatic and / or locally advanced cancer. In some embodiments of any of the methods of treating cancer, the cancer is inoperable.

[0038] Definitions As used herein, the term "administer" refers to the administration of a composition (e.g., a compound or a salt or preparation thereof including a salt as described herein) to a subject or system. Administration to an animal subject (e.g., to a human) can be by any suitable route. For example, in some embodiments, administration is by bronchial (including by bronchial instillation), buccal, enteral, interdermal, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by tracheal instillation), transdermal, vaginal, and vitreous.

[0039] "Biological sample" or "sample" means a fluid or solid sample from a subject. Biological samples can include cells; cellular, nucleic acid, protein, or membrane extracts; or body fluids including blood or (e.g., plasma, serum, saliva, urine, bile). Solid biological samples include samples taken from feces, rectum, central nervous system, bone, breast tissue, kidney tissue, cervix, endometrium, head and neck, gallbladder, parotid gland tissue, prostate, brain, pituitary gland, kidney tissue, muscle, esophagus, stomach, small intestine, colon, liver, spleen, pancreas, thyroid tissue, heart tissue, lung tissue, bladder, adipose tissue, lymph node tissue, uterus, ovarian tissue, adrenal tissue, testicular tissue, tonsils, and thymus. Liquid biological samples include samples taken from blood, serum, plasma, pancreatic juice, CSF, semen, prostatic fluid, seminal plasma, urine, saliva, sputum, mucus, bone marrow, lymph, and tears. Samples can be obtained by standard methods including, for example, venipuncture and surgical biopsy. In certain embodiments, the biological sample is a blood, plasma, or serum sample. In some embodiments, the biological sample is a tumor sample from a biopsy.

[0040] The term "cancer" refers to any cancer caused by the proliferation of malignant neoplastic cells, such as tumors, neoplasms, cell tumors, sarcomas, leukemias, and lymphomas.

[0041] "Determining the cell type level" means detecting the cell type by a method known in the art, either directly or indirectly. "Determining directly" means performing a process to obtain a physical entity or value (e.g., performing an assay or test on a sample, or "analyzing the sample" as the term is defined herein). "Determining indirectly" refers to receiving a physical entity or value from another entity or source (e.g., a third-party laboratory that has directly obtained the physical entity or value). Methods for measuring the cell level generally include, but are not limited to, flow cytometry and immunohistochemistry. Representative methods are provided herein. In some embodiments of any of the foregoing methods, the levels of MDSC and / or activated T cells can be determined as described in Iclozan et al., Cancer Immunol. Immunother. 2013, 62(5):909-918. In some embodiments of any of the foregoing methods, the levels of MDSC and / or activated T cells can be determined as described in Kitano et al., Cancer Immunol. Res. 2014, 2(8);812-821.

[0042] A cancer "determined to be drug-resistant", as used herein, refers to a cancer that is drug-resistant based on non-responsiveness or reduced responsiveness to a chemotherapeutic agent, or a cancer that is expected to be drug-resistant based on a prognostic assay (e.g., a gene expression assay).

[0043] A "drug-resistant" cancer means a cancer that does not respond or exhibits reduced responsiveness to one or more chemotherapeutic agents (e.g., any of the agents described herein).

[0044] The term "effective amount" means an amount sufficient to treat a disease, disorder, and / or condition when administered to a population suffering from or susceptible to that disease, disorder, and / or condition in accordance with a therapeutic agent dosing regimen. In some embodiments, a therapeutically effective amount is one that reduces the incidence and / or severity of and / or delays the onset of one or more symptoms of a disease, disorder, and / or condition. One of ordinary skill in the art understands that the term "effective amount" does not require that an actually successful treatment be achieved in a particular individual. Rather, an effective amount can be an amount that provides a particular desired pharmacological response in a significant number of subjects when administered to patients in need of such treatment. It is specifically understood that a particular subject can be "non-responsive" to an "effective amount." By way of example only, a non-responsive subject can have low bioavailability such that clinical effectiveness is not achieved. In some embodiments, reference to an effective amount can be reference to an amount as measured in one or more specific tissues (e.g., tissues affected by a disease, disorder, or condition) or body fluids (e.g., blood, saliva, serum, sweat, tears, urine). One of ordinary skill in the art understands that in some embodiments, an effective amount can be formulated and / or administered as a single dose. In some embodiments, an effective amount can be formulated and / or administered in multiple doses, e.g., as part of a dosing regimen.

[0045] As used herein, the terms "not responsive to previous treatment" or "previous treatment was ineffective" refer to cancer that has progressed despite treatment with that therapy.

[0046] "Level" means the level of a cell type when compared to a reference value. The reference value can be any useful reference value as defined herein. "Level decrease" or "level increase" of a cell type means a decrease or increase in the cell level when compared to the reference value (e.g., a decrease or increase of about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500% or more when compared to the reference value; a decrease or increase of more than about 10%, about 15%, about 20%, about 50%, about 75%, about 100% or about 200%; a decrease or increase of less than about 0.01-fold, about 0.02-fold, about 0.1-fold, about 0.3-fold, about 0.5-fold, about 0.8-fold or less; or an increase of about 1.2-fold, about 1.4-fold, about 1.5-fold, about 1.8-fold, about 2.0-fold, about 3.0-fold, about 3.5-fold, about 4.5-fold, about 5.0-fold, about 10-fold, about 15-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 100-fold, about 1000-fold or more). The level of a cell type can be expressed in mass / vol (e.g., g / dL, mg / mL, μg / mL, ng / mL) or % units relative to the total cells in the sample. In some embodiments of any of the foregoing methods, the reference value is a sample from a healthy subject such as a subject without cancer. In some embodiments of any of the foregoing methods, the reference value is an artificial sample having a level (e.g., the level of MDSC or activated T cells such as monocytes and / or granulocytic MDSC) that has been shown to be beneficial in the treatment of a disorder.

[0047] As used herein, "metastatic nodule" refers to an aggregation of tumor cells in the body at a site other than the original tumor site.

[0048] As used herein, "metastatic tumor" refers to a tumor or cancer that forms cancer cells that are likely to or have begun to metastasize or spread from one location within a subject to another location, either through the lymphatic system or through hematogenous spread, for example, causing a secondary tumor within the subject. Such metastatic behavior can indicate a malignant tumor. In some cases, metastatic behavior may be associated with enhanced cell migration and / or invasion behavior of tumor cells.

[0049] Examples of cancers that can be defined as metastatic include, but are not limited to, lung cancer (e.g., non-small cell lung cancer), breast cancer, ovarian cancer, colorectal cancer, cholangiocarcinoma, bladder cancer, brain cancers including glioblastoma and medulloblastoma, cervical cancer, choriocarcinoma, endometrial cancer, esophageal cancer, gastric cancer, hematological neoplasms, multiple myeloma, leukemia, intraepithelial neoplasms, liver cancer, lymphoma, neuroblastoma, oral cancer, pancreatic cancer, prostate cancer, sarcoma, skin cancers including melanoma, basal cell carcinoma, squamous cell carcinoma, testicular tumors, stromal tumors, germ cell carcinomas, thyroid cancer, and kidney cancer.

[0050] "Non-metastatic cell migration cancer" as used herein refers to a cancer that does not migrate through the lymphatic system or through hematogenous spread.

[0051] As used herein, the term "pharmaceutical composition" refers to an active compound, or a pharmaceutically acceptable salt thereof, formulated together with one or more pharmaceutically acceptable carriers. In some embodiments, the active compound or salt is present in a unit dosage amount appropriate for administration in a treatment regimen that demonstrates a statistically significant likelihood of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, the pharmaceutical composition can be formulated specifically for administration in solid or liquid form, including, but not limited to: oral administration, such as aqueous or non-aqueous solutions or suspensions, such as tablets, boluses, powders, granules, pastes for application to the tongue, formulated for buccal, sublingual, and systemic absorption; parenteral administration, such as by subcutaneous, intramuscular, intravenous, or epidural injection, such as sterile solutions or suspensions or sustained release formulations; topical application, such as as creams, ointments, or controlled release patches, or sprays applied to the skin, lung, or oral cavity; intravaginal or rectal, such as as pessaries, creams, or foams; sublingual; ocular; transdermal; or nasal, pulmonary, and other mucosal surfaces.

[0052] "Pharmaceutically acceptable excipient" as used herein refers to any inert ingredient (e.g., a vehicle capable of suspending or dissolving the active compound) having properties that are non-toxic and non-inflammatory in a subject. Typical excipients include, for example: anti-adhesives, antioxidants, binders, coating agents, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film-forming or coating agents, flavors, fragrances, glidants (flow promoters), lubricants, preservatives, printing inks, adsorbents, suspending or dispersing agents, sweeteners, or water of hydration.

[0053] As used herein, the term "pharmaceutically acceptable salt" refers to salts of the compounds described herein that are within the scope of sound medical judgment, are free of undue toxicity, irritation, allergic response, etc., are suitable for use in contact with human and animal tissues, and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977, and Handbook of Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. The salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reacting the free base with a suitable organic acid.

[0054] As used herein, "progression-free survival" refers to the length of time during and after dosing or treatment in which there is no progression of the disease being treated (e.g., cancer).

[0055] As used herein, "proliferation" includes the replication or growth of similar forms (cells) due to constituent (cell) elements.

[0056] As used herein, "retarding metastatic spread" means decreasing or stopping the formation of new sites; or decreasing, stopping or reversing the tumor burden.

[0057] As used herein, the term "subject" refers to a human or non-human animal (e.g., a mammal such as a non-human primate, horse, cow or dog).

[0058] The term "substantially" refers to a quantitative state representing the whole or nearly the whole extent or degree of a feature or characteristic of interest. One of ordinary skill in the art of biology understands that biological and chemical phenomena rarely, if ever, reach completion and / or progress to perfection or achieve or avoid absolute results. Thus, the term "substantially" is used herein to capture the potential lack of perfection inherent in many biological and chemical phenomena.

[0059] A "treatment plan" refers to a dosing plan in which administration across a relevant population correlates with a desired or beneficial treatment outcome.

[0060] The term "treatment" (as well as "treat" or "treating") in its broadest sense refers to the administration, in part or in whole, of a substance (such as a provided composition) that reduces, alleviates, relieves, inhibits, delays the onset of, reduces the severity of, and / or decreases the incidence of one or more symptoms, characteristics, and / or causes of a particular disease, disorder, and / or condition. In some embodiments, such treatment can be administered to a subject who does not exhibit symptoms of the relevant disease, disorder, and / or condition and / or who exhibits only early signs of the disease, disorder, and / or condition. Alternatively or additionally, in some embodiments, treatment can be administered to a subject who exhibits one or more established signs of the relevant disease, disorder, and / or condition. In some embodiments, treatment can be of a subject diagnosed as having the relevant disease, disorder, and / or condition. In some embodiments, treatment can be of a subject known to have one or more susceptibility factors that are statistically correlated with an increased risk of developing the relevant disease, disorder, and / or condition.

[0061] As used herein, the term "PD-1 inhibitor" refers to a compound such as an antibody that can inhibit the activity of the protein encoded by the PDCD1 gene in humans. Known PD-1 inhibitors include nivolumab, pembrolizumab, pidilizumab, BMS 936559, and atezolizumab.

[0062] As used herein, the term "PD-L1 inhibitor" refers to a compound such as an antibody that can inhibit the activity of the protein encoded by the CD274 gene in humans. Known PD-L1 inhibitors include atezolizumab and durvalumab.

[0063] As used herein, the term "CTLA-4 inhibitor" refers to a compound such as an antibody that can inhibit the activity of the protein encoded by the CTLA4 gene in humans. A known CTLA-4 inhibitor is ipilimumab.

[0064] As used herein, the term "CSF-1R inhibitor" means a compound such as an antibody that can inhibit the activity of the protein encoded by the CSF1R gene in humans. Known CSF-1R inhibitors include pexidartinib and 4-(2,4-difluoroanilino)-7-ethoxy-6-(4-methylpiperazin-1-yl)quinoline-3-carboxamide.

[0065] As used herein, the term "IDO inhibitor" refers to a compound such as an antibody that can inhibit the activity of the protein encoded by the IDO1 gene in humans. Known IDO inhibitors include norharman, rosmarinic acid, and alpha-methyl-tryptophan.

[0066] The term "A1 adenosine inhibitor" as used herein refers to a compound such as an antibody that can inhibit the activity of the protein encoded by the ADORA1 gene in humans. Known A1 adenosine inhibitors include 8-cyclopentyl-1,3-dimethylxanthine, 8-cyclopentyl-1,3-dipropylxanthine, 8-phenyl-1,3-dipropylxanthine, bambifylline, BG-9719, tonophrine, FK-453, FK-838, rolofylline, and N-0861.

[0067] The term "A2A adenosine inhibitor" as used herein refers to a compound such as an antibody that can inhibit the activity of the protein encoded by the ADORA2A gene in humans. Known A2A adenosine inhibitors include ATL-4444, istradefylline, MSX-3, preladenant, SCH-58261, SCH-412,348, SCH-442,416, 2-butyl-9-methyl-8-(triazol-2-yl)purin-6-amine, VER-6623, VER-6947, VER-7835, viadenant, and ZM-241,385.

[0068] As used herein, the term "A2B adenosine inhibitor" means a compound such as an antibody that can inhibit the activity of the protein encoded by the ADORA2B gene in humans. Known A2B adenosine inhibitors include N-[5-(1-cyclopropyl-2,6-dioxo-3-propyl-7H-purin-8-yl)pyridin-2-yl]-N-ethylpyridine-3-carboxamide, 3-ethyl-1-propyl-8-[1-[[3-(trifluoromethyl)phenyl]methyl]pyrazol-4-yl]-7H-purine-2,6-dione, MRS-1706, MRS-1754, N-[2-[[2-phenyl-6-[4-(3-phenylpropyl)piperazine-1-carbonyl]-7H-pyrrolo[2,3-d]pyrimidin-4-yl]amino]ethyl]acetamide, PSB-603, PSB-0788, and PSB-1115.

[0069] As used herein, the term "A3A adenosine inhibitor" refers to a compound such as an antibody that can inhibit the activity of the protein encoded by the ADORA3 gene in humans. Known A3A adenosine inhibitors include KF-26777, MRS-545, MRS-1191, MRS-1220, MRS-1334, MRS-1523, MRS-3777, MRE-3005-F20, MRE-3008-F20, PSB-11, OT-7999, VUF-5574, and SSR161421.

[0070] As used herein, the term "arginase inhibitor" refers to a compound that can inhibit the activity of the protein encoded by the ARG1 or ARG2 gene in humans. Known arginase inhibitors include (2s)-(+)-amino-5-iodoacetamidopentanoic acid, NG-hydroxy-L-arginine, (2S)-(+)-amino-6-iodoacetamidohexanoic acid, and (R)-2-amino-6-borono-2-(2-(piperidin-1-yl)ethyl)hexanoic acid.

[0071] As used herein, the term "HDAC inhibitor" refers to a compound such as an antibody that can inhibit the activity of proteins that are members of the histone deacetylase class of enzymes, such as HDAC1, HDAC2, HDAC3, HDAC4, HDAC5, HDAC6, HDAC7, HDAC8, HDAC9, HDAC10, HDAC11, SIRT1, SIRT2, SIRT3, SIRT4, SIRT5, SIRT6, and SIRT7. Known HDAC inhibitors include valproic acid, SAHA, and romidepsin.

[0072] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials are described herein for use in the present disclosure; other suitable methods and materials known in the art may also be used. The materials, methods, and examples are illustrative only and not limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present application, including definitions, will control.

[0073] Details of one or more embodiments of the invention are set forth in the following description. Other features, objects, and advantages of the invention will become apparent from the description and claims.

Brief Description of the Drawings

[0074]

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[0075] To identify salts of 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]methyl-(2,2-diphenylethyl)amino]butoxy]phenyl]acetic acid (Compound 1) having improved processability and stability, which are LXR agonists, the inventors of the present invention conducted extensive salt screening experiments using over 65 counterion sources and a solvent system prepared from over 15 solvents or solvent mixtures.

[0076] In an initial screen of common counterion sources, the only salts that appeared to be prepared in crystalline form were those prepared with hydrochloric acid, DL-mandelic acid, and naphthalenesulfonic acid, and the hydrochloride salt was the only salt that could be reproduced and scaled up. However, it was found that the hydrochloride salt, along with several strong acid salts (i.e., H 2 SO 4 , HBr, p-toluenesulfonic acid, and methanesulfonate), tended to be unstable and were found to readily undergo apparent esterification in an alcohol solvent, for example. Subsequent studies on salts prepared with weak acids, such as oleic acid, octanoic acid, and acetic acid, resulted in the isolation of only the free compound.

[0077] Furthermore, the inventors have found that the hydrochloride salt is not stable under vacuum and at medium to high temperatures (e.g., above 50 °C). Under these conditions, the hydrochloride salt tends to discolor and / or tends to lose hydrochloric acid. The inventors have also found that extracting the hydrochloride salt with water causes loss of hydrochloric acid and further complicates processing. The hydrochloride salt is further found to undergo apparent esterification in the presence of free alcohol groups, for example, in an alcohol solvent or in a formulation containing excipients having free alcohol groups such as sorbitol, or esterification in the presence of lipophilic esters such as linoleic acid esters. Due to these instability problems of the hydrochloride salt, it becomes less suitable for processing into a drug product, and thus, salt screening studies were conducted to find more suitable salt forms.

[0078] Subsequent salt screening studies were then carried out to identify counterions and solvent systems that would assist in the formation of salts with improved physical properties and stability. From these salt screening studies, including experiments using over 65 different counterion sources, the 2:1 (Compound 1:zinc) salt was unexpectedly one of the few salts that formed as a stable solid against gels, gum substances, oils, and semi-solids formed by a large number of counterion sources. The 2:1 (Compound 1:zinc) salt, as a reproducible flowable solid, is quite suitable for scale-up and pharmaceutical development.

[0079] 2-[3-[(3R)-3-[[2-Chloro-3-(trifluoromethyl)phenyl]methyl-(2,2-diphenylethyl)amino]butoxy]phenyl]acetic acid 2-[3-[(3R)-3-[[2-Chloro-3-(trifluoromethyl)phenyl]methyl-(2,2-diphenylethyl)amino]butoxy]phenyl]acetic acid (Compound 1) has the following structure:

Chemical formula

[0080] In syngeneic and human xenograft mouse tumor models of melanoma (with different genetic backgrounds), glioblastoma, TNBC, ovarian cancer, and lung cancer, compound 1 inhibited primary tumor growth by 48 - 95%. The events of tumor growth inhibition varied by model. In a mouse model of TNBC metastasis, compound 1 inhibited the spread of cancer cell metastasis by about 9-fold. Furthermore, the antitumor activity of compound 1 in combination with an anti-PD-1 antibody inhibited tumor growth by >80% in a syngeneic mouse melanoma model that would not respond to the anti-PD-1 antibody in another way. Additionally, the inhibition of tumor growth in the same syngeneic mouse melanoma model was superior when the mice received combination therapy with compound 1 and an anti-CTLA-4 antibody compared to either therapy alone. Similarly, in a syngeneic mouse melanoma model, compound 1 showed superior antitumor efficacy (>80%) when combined with dacarbazine compared to either treatment alone. In tumor growth studies, the minimum effective dose ranged from 25 - 40 mg / kg / day when administered orally (PO), resulting in exposures in the range of 10,000 - 50,000 ng-h / mL.

[0081] In the safety pharmacology evaluation, compound 1 did not inhibit the channel conductance of the human ether-à-go-go-related gene (hERG) in the in vitro hERG assay, but a significant increase occurred. There was no effect of compound 1 on the qualitative electrocardiogram (ECG) parameters (PR or QTc interval or QRS duration) in dogs, but after administration at 150 (reduced to 100) mg / kg / day, there was a dose-related decrease in the mean heart rate at intervals on the first day after administration, which was significantly different in females. This change was not confirmed during the recovery period and was not considered harmful. Furthermore, no adverse effects were confirmed in rats during the Functional Observation Battery (FOB) or respiratory evaluation. Assuming the favorable safety profile of compound 1 at the highest dose tested in the repeated-dose toxicity study, the potential for cardiovascular, respiratory, or central nervous system (CNS) effects is considered low.

[0082] In the oral PK study, compound 1 was well absorbed in CD-1 mice, with an absolute calculated oral bioavailability (%F) often exceeding 100%, indicating possible enterohepatic recirculation of the parent compound. The time to reach the maximum plasma concentration (Tmax) was approximately the same in male and female mice and ranged from 2 to 8 hours. The apparent mean oral half-life (t1 / 2) ranged from 6.5 to 8 hours in mice. When administered to mice with food, there was a significant food effect, as shown by compound 1 plasma concentrations being >2-fold higher. In Sprague-Dawley (SD) rats, the total mean F (%) (after a 30 mg / kg oral dose of 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]methyl-(2,2-diphenylethyl)amino]butoxy]phenyl]acetic acid) was moderate (about 31%), Tmax was approximately the same in male and female rats and ranged from 4 to 8 hours. Compound 1 was cleared at a lower rate in female rats compared to males, resulting in higher systemic exposure to compound 1 in females at all tested dose levels. The apparent mean oral t1 / 2 in female rats was 6.5 hours (not calculable in males). In male Beagle dogs orally administered compound 1, Tmax ranged from 4 to 8 hours. The mean F (%) was moderate (18 - 30% depending on dose and formulation) and the apparent mean t1 / 2 ranged from 5 to 6.7 hours. In cynomolgus monkeys, the mean F (%) was low to moderate (6 - 19% depending on dose and formulation). After oral administration, the monkeys had a mean Tmax of 4 hours. The mean oral t1 / 2 ranged from 5.5 to 8 hours.

[0083] Compound 1 is subject to Phase I and Phase II metabolism, including oxidation, dealkylation, glucuronidation, and combinations thereof. In vitro, Compound 1 is mainly metabolized by cytochrome P450 (CYP) isoform CYP3A4, but is also a substrate for CYP2E1, CYP2C9, CYP2C19, and possibly CYP2J2. Compound 1 is not a strong inhibitor of any human CYP450 in vitro, while it is a moderate inhibitor of CYP2C8 (50% inhibitory concentration [IC50] 7.5 μM) and a weak inhibitor of 2B6 (IC50 15 μM). Compound 1 weakly inhibited 1A1, 2A6, 2C9, 2C19, 2D6, 2E1, and 3A4 in vitro, but time-dependent inhibition (TDI) of CYP3A was demonstrated in vitro using testosterone as a substrate. Induction of CYP3A by Compound 1 was demonstrated in primary cultures of cryopreserved hepatocytes (two donors), and the possibility of Compound 1 inducing CYP2B6 cannot be excluded (induced more than two-fold at 1 / 3 the concentration of one donor). Compound 1 did not induce CYP1A2. In efflux transporters, Compound 1 does not inhibit P-glycoprotein (P-gp), but inhibits breast cancer resistance protein (BCRP) transport in vitro (55% at 5 μM). Compound 1 is a potent inhibitor of the uptake transporter organic anion transporting polypeptide (OATP) 1B1 in vitro (IC50 0.099 μM). Compound 1 also appears to be a moderate inhibitor of OATP1B3 (IC50 3.7 μM). Compound 1 weakly inhibited OAT1, OAT3, and OCT2 in vitro at an inhibition rate of less than 50% at 50 μM.

[0084] Potential risks of Compound 1 in a clinical setting based on animal toxicology studies include increased serum cholesterol and TG, neutropenia / leukopenia, nausea and / or vomiting, elevated liver enzymes, onset or worsening of cataracts, cardiac arrhythmia and / or decreased cardiac function, adenocarcinoma of the harder gland, and / or generalized edema.

[0085] Nivolumab Nivolumab is a fully human immunoglobulin (Ig) G4 monoclonal antibody against the negative immune regulatory system human cell surface receptor programmed death-1 (PD-1), which has immune checkpoint inhibition and anti-tumor activity. Nivolumab binds to PD-1, an Ig superfamily transmembrane protein, by its ligands, programmed cell death ligand 1 (PD-L1), which is overexpressed on certain cancer cells, and programmed cell death ligand 2 (PD-L2), which is mainly expressed on antigen-presenting cells, and blocks its activation. As a result, T cells are activated and a cellular immune response against tumor cells or pathogens occurs. Activated PD-1 negatively regulates T cell activation and plays an important role in tumor evasion from host immunity. The nivolumab dose would be 240 mg administered as an intravenous infusion over 60 minutes on days 1 and 15 of each 28-day cycle. There is a possibility that the toxicities of Compound 1 and nivolumab may overlap. Specifically, grade 4 neutropenia DLT has been seen in monotherapy with Compound 1, and myelosuppression can be confirmed with nivolumab treatment. The pharmacological actions of Compound 1 include the regulation of steroid biosynthesis. As a result, hyperlipidemia has been confirmed in subjects treated with Compound 1 and has also been reported in subjects treated with nivolumab. Biochemical abnormalities of liver function have been confirmed in the preclinical toxicity study of Compound 1, and immune-mediated hepatitis has been confirmed with nivolumab.

[0086] Ipilimumab Ipilimumab is a recombinant human IgG1 kappa monoclonal antibody that binds to cytotoxic T lymphocyte protein 4 (CTLA-4). CTLA-4 is a negative regulator of T cell activity. By binding to CTLA-4, ipilimumab blocks the interaction of its ligand, CD80 / CD86, with CTLA-4. Blockade of CTLA-4 has been shown to enhance T cell activation and proliferation, such as the activation and proliferation of tumor-infiltrating effector T cells. Inhibition of CTLA-4 signaling can also reduce regulatory T cell function, which can contribute to a general increase in T cell responsiveness, such as an anti-tumor immune response.

[0087] In some embodiments, the ipilimumab dosage is 3 mg / kg administered as an intravenous infusion on day 1 of each 28-day cycle for up to 4 administrations.

[0088] There is a potential for overlap in the toxicity of Compound 1 and ipilimumab. Biochemical abnormalities of liver function were confirmed in preclinical toxicity studies with Compound 1; immune-mediated hepatitis was confirmed with ipilimumab treatment.

[0089] Docetaxel Docetaxel is an anti-tumor agent belonging to the taxoid family. It is manufactured by semi-synthesis starting from a precursor extracted from the renewable needle biomass of yew plants. The chemical name of docetaxel is (2R,3S)-N-carboxy-3-phenylisoserine, N-t-butyl ester, 13-ester with 5b-20-epoxy-1,2a,4,7b,10b,13a-hexahydroxytax-11-en-9-one 4-acetate 2-benzoate trihydrate.

[0090] In some embodiments, docetaxel is administered as an intravenous infusion on days 1, 8, and 15 of each 28-day cycle. In some embodiments, the docetaxel dosage is 35 mg / m 2 It was. In some embodiments, 28 mg / m 2 is the docetaxel dosage.

[0091] There is a potential for overlap in the toxicity of Compound 1 and docetaxel. Specifically, grade 4 neutropenia DLT has been seen in monotherapy with Compound 1, and myelosuppression can be confirmed with docetaxel treatment. Biochemical abnormalities of liver function have been confirmed in preclinical toxicity studies with Compound 1; hepatotoxicity has been confirmed with docetaxel treatment.

[0092] Pembrolizumab Pembrolizumab is a programmed cell death receptor-1 (PD1) blocking antibody. Pembrolizumab is a humanized monoclonal IgG4 kappa antibody with an approximate molecular weight of 149 kDa. Pembrolizumab is produced in recombinant Chinese hamster ovary (CHO) cells.

[0093] In some embodiments, after all procedures and evaluations are completed, and before the administration of other drugs, and with a 30-minute interval between the administrations of the next drug, pembrolizumab is administered at a dose of 200 mg by intravenous infusion for 30 minutes on day 1 of each 21-day cycle.

[0094] Carboplatin The chemical name of carboplatin (United States Pharmacopeia) is platinum diamine [1,1-cyclobutanedicarboxylato(2-)-O,O']-(SP-4-2). Carboplatin (United States Pharmacopeia) is a crystalline powder. It is water-soluble at a concentration of about 14 mg / mL, and the pH of a 1% solution is 5 - 7. It is substantially insoluble in ethanol, acetone, and dimethylacetamide. Carboplatin produces mainly interstrand DNA cross-links rather than DNA-protein cross-links. This action appears to be cell cycle non-specific. Carboplatin induces an equal number of drug-DNA cross-links, resulting in corresponding lesions and biological effects.

[0095] In some embodiments, the initial dose of carboplatin infusion is determined using a formula based on the subject's existing renal function or renal function and the desired platelet nadir (renal excretion is the main route of carboplatin excretion). By using a dosing formula as compared to empirical dose calculations based on body surface area, it is possible to compensate for differences in the subject's pretreatment renal function such that otherwise, dosing may be insufficient (in subjects with the above-average renal function) or may result in overdose (in subjects with impaired renal function).

[0096] A simple formula for the dosage based on the glomerular filtration rate (GFR (mL / min)) of the subject and the area under the concentration-time target curve (AUC (mg / mL·min)) of carboplatin infusion was proposed by Calvert. In these studies, GFR was measured by Cr-EDTA clearance. The Calvert formula for carboplatin administration is as follows: Total dose (mg) = (target AUC) × (GFR + 25)

[0097] Note that using this formula, the total dose of carboplatin is calculated in mg, not mg / m 2 A target AUC of 4 - 6 mg / mL·min using single-agent carboplatin is thought to provide the most appropriate dosage range in pre-treated subjects. This study also showed a trend between the AUC of single-agent carboplatin administered to pre-treated subjects and the potential for toxicity.

[0098] Pemetrexed Pemetrexed (for injection) is a folic acid analogue metabolic inhibitor. The drug substance, pemetrexed disodium heptahydrate, has the chemical name L-glutamic acid, N-[4-[2-(2-amino-4,7-dihydro-4-oxo-1H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]-disodium salt heptahydrate with the molecular formula C 20 H 19 N 5 Na 2 O 6 ·7H 2 O and a molecular weight of 597.49 g / mol.

[0099] Pemetrexed exerts its antitumor activity by inhibiting folate-dependent metabolic processes essential for cell replication. From in vitro studies, it has been found that pemetrexed acts as a multitargeted folate antagonist by inhibiting thymidylate synthase (TS), dihydrofolate reductase (DHFR), and glycineamide ribonucleotide formyltransferase (GARFT), which are important for de novo biosynthesis of thymidine and purine nucleotides. The pemetrexed drug action in malignant cells becomes persistent by polyglutamylating the metabolites of pemetrexed that have a persistent intracellular half-life.

[0100] In some embodiments, the pemetrexed dosage is 500 mg / m on the first day of each 21-day cycle for up to 4 cycles. 2 In some embodiments, subjects treated with pemetrexed must be instructed to receive folic acid and vitamin B12 as prophylactic measures to reduce treatment-related hematological and GI toxicities. In some embodiments, corticosteroids may also be prescribed to the subject to be taken twice daily for 3 days starting on the day prior to each treatment with pemetrexed.

[0101] There may be overlapping toxicities among Compound 1, pemetrexed, and carboplatin. Specifically, grade 4 neutropenia DLT has been seen in monotherapy with Compound 1, and myelosuppression can be confirmed in combination therapy of Compound 1 and carboplatin with pemetrexed.

[0102] Adverse Event An adverse event (AE) is any undesirable medical occurrence in a subject or clinical subject who has received a pharmaceutical product and does not necessarily have to have a causal relationship with this treatment. Thus, whether related to the investigational product or not, an AE can be an undesirable and unintended sign (such as abnormal laboratory findings), symptom, or disease that temporarily accompanies the use of the investigational product.

[0103] Death and disease progression (PD) are not considered AEs. Death is considered the result of one or more major AEs, and PD is considered the worsening of the underlying disease. Pre-existing conditions (existing prior to the start of the AE collection period) are considered the current medical condition and are not considered AEs. However, a worsening or complication of such a current medical condition The disease is is an AE.

[0104] An AE or suspected adverse reaction is considered severe if it results in death; is life-threatening, i.e., the subject has an immediate risk of death from the reaction when it occurs, even if it occurs in a more severe form; requires hospitalization of the subject or prolongation of the current hospitalization; results in persistent or significant physical disability / incapacity; is a congenital malformation / congenital anomaly; or is a significant medical event, but does not include a reaction that might hypothetically cause death.

[0105] Method of treatment Cancer can be treated using the methods described herein.

[0106] As a result of cancer treatment, a reduction in the size or volume of the tumor can occur. For example, after treatment, the tumor size can shrink by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) compared to its size before treatment. The tumor size can be measured by any reproducible measurement means. The tumor size can be measured as the diameter of the tumor or by any reproducible measurement means.

[0107] As a further result of cancer treatment, a reduction in the number of tumors can occur. For example, after treatment, the number of tumors can decrease by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) compared to the number before treatment. The number of tumors can be measured by any reproducible measurement means. The number of tumors can be measured by counting the tumors visible to the naked eye or at a specified magnification (e.g., 2×, 3×, 4×, 5×, 10× or 50×).

[0108] As a result of treating cancer, the number of metastatic nodules in other tissues or organs far from the primary tumor site can be reduced. For example, after treatment, the number of metastatic nodules is reduced by 5% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) compared to the number before treatment. The number of metastatic nodules can be measured by some reproducible measuring means. The number of metastatic nodules can be measured by counting the metastatic nodules visible to the naked eye or at a specified magnification (e.g., 2×, 10× or 50×).

[0109] As a result of treating cancer, the average survival time of a population of subjects treated according to the present invention can be extended compared to a population of untreated controls. For example, the average survival time is extended by more than 30 days (more than 60 days, 90 days or 120 days). The extension of the average survival time of the population can be measured by some reproducible means. The extension of the average survival time of the population can be measured, for example, by calculating the average length of survival after the start of treatment with the compound of the present invention for the population. The extension of the average survival time of the population can also be measured, for example, by calculating the average length of survival after the completion of the first round of treatment with the compound of the present invention for the population.

[0110] As a result of treating cancer, also, the mortality rate of a population of treated subjects can be decreased compared to an untreated population. For example, the mortality rate is decreased by more than 2% (e.g., more than 5%, 10% or 25%). The decrease in the mortality rate of the population of treated subjects can be measured by some reproducible means, for example, by calculating the average number of disease-related deaths per unit time after the start of treatment with the compound of the present invention for the population. The decrease in the mortality rate of the population can also be measured, for example, by calculating the average number of disease-related deaths per unit time after the completion of the first round of treatment with the compound of the present invention for the population.

[0111] As a result of cancer treatment, the average progression-free survival time of the treated population can also be extended compared to the untreated population. For example, the average progression-free survival time is extended by more than 30 days (more than 60 days, 90 days, or 120 days). The extension of the average progression-free survival time of the population can be measured by any reproducible means. The extension of the average progression-free survival time of the population can be measured, for example, by calculating the average length of the progression-free survival rate for the population after the start of treatment with the compound of the present invention. The extension of the average progression-free survival time of the population can also be measured, for example, by calculating the average length of progression-free survival for the population after completion of the first round of treatment with the compound of the present invention.

[0112] Composition Compositions containing a suitable carrier and one or more of the above-mentioned therapeutic agents are within the scope of the present invention.

[0113] As described above, the pharmaceutical composition of the present invention further comprises any and all solvents, diluents, or other liquid excipients, dispersions or suspensions, acids, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, and lubricants suitable for the desired specific dosage form, etc., pharmaceutically acceptable excipients used herein. Remington’s Pharmaceutical Sciences, Sixteenth Edition, E.W. Martin (Mack Publishing Co., Easton, Pa., 1980) discloses various excipients used in the formulation of pharmaceutical compositions and known techniques for their manufacture. For example, as long as the compound of the present invention is not incompatible with conventional excipients by producing any undesirable biological effects or interacting detrimentally with other components of the pharmaceutical composition, its use is contemplated to be within the scope of the present invention.

[0114] The above compositions in any of the above forms can be used for the treatment of cancer, or any other disease or medical condition described herein. An effective amount means the amount of the active compound / agent necessary to confer a therapeutic effect on the subject being treated. The effective dosage will vary depending on the type of disease being treated, the route of administration, the use of excipients, and the possibility of co-use with other therapeutic treatments, as will be appreciated by those skilled in the art.

[0115] The pharmaceutical compositions of the present invention can be administered parenterally, orally, nasally, rectally, topically, or buccally. As used herein, the term "parenteral" means subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intra-arterial, intrasynovial, intrasternal, intramedullary, intralesional, or intracranial injection, and any suitable injection technique.

[0116] Injectable sterile compositions can be solutions or suspensions in parenterally acceptable non-toxic diluents or solvents.

[0117] Compositions for oral administration can be in any orally acceptable dosage form, such as capsules, tablets, emulsions, and aqueous suspensions, dispersions, and solutions. When an aqueous suspension or emulsion is administered orally, the active ingredient can be suspended or dissolved in an oil phase combined with an emulsifying or suspending agent. If desired, certain sweetening, flavoring, or coloring agents can be added.

[0118] The pharmaceutical compositions for topical administration according to the present invention described can be formulated as solutions, ointments, creams, suspensions, lotions, powders, pastes, gels, sprays, aerosols or oils. Alternatively, the topical formulation can take the form of a patch or dressing impregnated with the active ingredient, optionally containing one or more excipients or diluents. In some preferred embodiments, the topical formulation contains substances that will enhance the absorption or penetration of the agent through the skin or other affected area.

[0119] Combination therapy In some embodiments of the methods described herein, the pharmaceutical composition may further comprise an additional compound having antiproliferative activity.

[0120] It is also understood that the compounds and pharmaceutical compositions of the present invention can be formulated and used in combination therapies, i.e., the compounds and pharmaceutical compositions can be formulated with one or more other desired therapeutic agents or medical procedures (e.g., surgery and / or radiation therapy), or administered simultaneously with, before, or after one or more other desired therapeutic agents or medical procedures. The specific combination of therapies (therapeutic agents or procedures) to be used in a combination regimen takes into account the compatibility of the desired therapeutic agent and / or procedure and the desired therapeutic effect to be achieved. It will also be understood that the therapies used can achieve the desired effect for the same disorder or that they can achieve different effects (e.g., control of some adverse effect).

Examples

[0121] Example 1, Method X-ray powder diffraction (XRPD) Method 1 XRPD analysis was performed using a PANalytical X’pert pro, and the sample was scanned at 3 - 35° 2θ. The material was gently ground to break up agglomerates and placed on a multiwell plate together with a Mylar polymer film to support the sample. The multiwell plate was then placed on the diffractometer and analyzed using a Cu K line (α1 λ = 1.54060 Å; α2 = 1.54443 Å; β = 1.39225 Å; α1:α2 ratio = 0.5) operating in the transmission mode (step size 0.0130° 2θ) with a 40 kV / 40 mA generator setting.

[0122] Method 2 An X-ray powder diffraction pattern was obtained using a Bruker D8 Advance equipped with a Cu Kα radiation source (λ = 1.54 Å), a 9-position sample holder, and a LYNXEYE ultra-fast detector. The sample was placed on a zero-background, silicon plate holder.

[0123] Polarizing microscope (PLM) Method 1 The presence of birefringence was determined using an Olympus BX53 polarized microscope equipped with a Motic camera and image capture software (Motic Images Plus 3.0). All images were recorded using a 10× or 20× objective lens.

[0124] Method 2 The sample was analyzed using an Olympus BX53 polarized microscope equipped with a PAXcam 3 digital microscope camera.

[0125] Thermogravimetric analysis / differential thermal analysis (TG / DTA) Method 1 Approximately 5 mg of the material was weighed into an open aluminum pan, placed on a simultaneous thermogravimetric analysis / differential thermal analysis apparatus (TG / DTA), and held at room temperature. Next, under a nitrogen purge, the sample was heated from 20 °C to 300 °C at a rate of 10 °C / min, and during that time, the change in sample weight was recorded along with any differential thermal events (DTA).

[0126] Method 2 TGA data were collected using a TA Instruments TGA Q500. Usually, the sample (about 10 mg) was placed in a pre-tared open aluminum sample pan and scanned from 25 °C to 300 °C at a rate of 10 °C / min using a nitrogen purge at 60 mL / min.

[0127] Differential scanning calorimetry (DSC) Method 1 Approximately 5 mg of the substance was weighed into an aluminum DSC pan and sealed with an aluminum lid. Then, the sample pan was placed on a TA Instruments Discovery DSC 2500 differential scanning calorimeter equipped with an RC90 cooler. Once a stable heat flow response was obtained, the sample and reference were heated / cooled under a nitrogen purge by using the following temperature program, and the resulting heat flow response was monitored. Program 1 · Heat from 20 °C to 260 °C at 10 °C / min and hold at 260 °C for 3 minutes; ·Cool from 260 °C to 20 °C at 10 °C / min and hold at 20 °C for 3 minutes; ·Heat from 20 °C to 260 °C at 10 °C / min and hold at 260 °C for 3 minutes; ·Cool from 260 °C to 20 °C at 10 °C / min and hold at 20 °C for 3 minutes; Program 2 ·Heat from 20 °C to 270 °C at 10 °C / min and hold at 270 °C for 3 minutes; ·Cool from 270 °C to 20 °C at 10 °C / min and hold at 20 °C for 3 minutes; ·Heat from 20 °C to 270 °C at 10 °C / min and hold at 270 °C for 3 minutes; ·Cool from 270 °C to 20 °C at 10 °C / min and hold at 20 °C for 3 minutes;

[0128] Method 2 DSC data were collected using a TA Instruments Q10 DSC. Usually, the sample (2 - 8 mg) was placed in an unsealed but covered hermetically alodined aluminum sample pan and scanned from 30 to 300 °C at a rate of 10 °C / min under a nitrogen purge of 50 mL / min.

[0129] 1 1H nuclear magnetic resonance spectroscopy ( 1 1H-NMR) Method 1 1H-NMR spectroscopic experiments were carried out on a Bruker AV500 (frequency: 500 MHz). 1 The experiments were performed in CDCl 3 3, and each sample was prepared to a concentration of approximately 10 mM.

[0130] Method 2 Samples were prepared by dissolving the compound in deuterated dimethyl sulfoxide containing 0.05% (v / v) tetramethylsilane (TMS). Spectra were collected at ambient temperature on a Bruker Avance 300 MHz NMR using TopSpin software. The number of scans was 16 for proton NMR.

[0131] 19 19F nuclear magnetic resonance spectroscopy ( 19 19F-NMR) Using a Bruker AV500 (frequency: 470 MHz) 19 F-NMR spectroscopic experiments were carried out. The experiments were performed in CDCl 3 and each sample was prepared at a concentration of approximately 10 mM.

[0132] High-performance liquid chromatography-ultraviolet detection (HPLC-UV) In the HPLC method, a C18 column is used with an acetonitrile / water / trifluoroacetic acid gradient.

[0133] Dynamic vapor sorption (DVS) The samples were analyzed using an Aquadyne DVS-2 gravimetric water sorption analyzer. The relative humidity was adjusted from 2 to 95%, and the weight of the samples was continuously monitored and recorded.

[0134] Karl Fisher (KF) The apparent moisture content in the samples was determined by Karl Fisher titration using a Mettler Toledo DL39 Coulometric KF titrator. HYDRANAL-Coulomat AD was used as the titrant. Approximately 20 mg of the solid was used for the titration. The analytical parameters are shown in the following table:

[0135]

Table 1

[0136] Fourier transform infrared spectroscopy (FTIR) FTIR analysis was performed using a Thermo Scientific NICOLE (trademark) IS (trademark) 10 FTIR spectrometer with the attenuated total reflection (ATR) method. Sufficient material was placed in the center of the spectrometer plate to obtain a spectrum.

[0137] Example 2 Salt Screening Study 1 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]-methyl-(2,2- diphenylethyl )amino]butoxy]phenyl]acetic acid salts, a first study was conducted. 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]-methyl-(2,2- diphenylethyl )amino]butoxy]phenyl]acetic acid salts, 26 counterion sources were selected for screening. The counterion sources are shown in Table 2 below:

[0138]

Table 2

[0139] Among the counterion sources investigated, crystalline salts were obtained from only three (hydrochloride, DL-mandelic acid, and naphthalenesulfonic acid), and the reproduction and scale-up of the three crystalline salts were successful only for the hydrochloride.

[0140] Subsequent studies revealed that the hydrochloride, as well as other salts prepared from strong acids such as H 2 SO 4 , HBr, p-toluenesulfonic acid, and methanesulfonic acid, were unstable. In particular, these salts react with the excipients and / or components of the formulation. This discovery led to studies on the use of weak acids such as oleic acid, octanoic acid, and acetic acid as counterion sources, none of which formed salts with 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]-methyl-(2,2- diphenylethyl )amino]butoxy]phenyl]acetic acid.

[0141] Study 2 A second study was conducted to identify salts that impart both improved processability and stability. The 34 counterion sources and 12 solvent conditions investigated in this study are summarized in Tables 3 and 4 below:

[0142]

Table 3

[0143]

Table 4

[0144] Evaluation of the initial solubility of amorphous compound 1 in 12 solvents / solvent mixtures was performed Dissolution was confirmed for most of the samples. All of cooling, addition of anti-solvent, aging, liquid-assisted grinding, pure grinding, and salt screening by evaporation were investigated. The isolated gummy substances were subjected to heating / cooling maturation cycles (room temperature / 50 °C), sonication, long-term storage under vacuum, long-term storage at 40 °C / 75% relative humidity, titration with anti-solvent, and liquid-assisted grinding (LAG) with water. Eight hits were obtained from a wide range of salt screening. Amorphous hits with high glass transition temperature (Tg) were identified from zinc chloride, naphthalene-2-sulfonic acid, 1,5-naphthalene disulfonic acid, and ethanesulfonic acid. One crystalline hit was identified from ethanesulfonic acid. Poorly crystalline materials were isolated from calcium hydroxide, magnesium hydroxide, and aluminum hydroxide.

[0145] Due to the limited characterization of the isolated hits, a focused screen was performed for the majority of promising counterions: calcium hydroxide, aluminum hydroxide, ethanesulfonic acid, and naphthalene-2-sulfonic acid. Calcium hydroxide and aluminum hydroxide were selected to evaluate whether the initial hits isolated via counterion exchange were single metal or mixed metal salts. The solids isolated from further screening matched the initial hits. From further screening, these samples appeared to be single metal salts. Since these hits were not mixed salts, a limited mixed metal screen was performed using magnesium hydroxide and calcium hydroxide. These attempts ended unsuccessfully. Ethanesulfonic acid and naphthalene-2-sulfonic acid were selected for further screening. However, the initial hits were not reproduced in further screening attempts. Amorphous solids were isolated, but their Tg was lower than that of both the free compound (32 °C) and the HCl salt (about 70 - 75 °C).

[0146] The zinc salt was isolated using the following procedure: Compound 1 (about 250 mg) was dissolved in THF (5 volumes) at room temperature. The solution was dispensed into an HPLC vial (75 μl, about 15 mg in solution). The solution was maintained at 50 °C for 5 minutes and observations were recorded. The sample was treated with 1.0 equivalent of sodium and then with the selected corresponding counterion (0.5 equivalent of zinc chloride). The sample was maintained at 50 °C for 10 minutes and then cooled to 5 °C at 0.1 °C / min. The resulting solution was maintained at 5 °C overnight and then stored at -20 °C. After 5 days, the sample remained in solution state and was evaporated under ambient conditions. A gummy substance was obtained after evaporation, which was treated with 10 volumes of heptane and subsequently sonicated for 2 hours. The collected solid was analyzed by XRPD and found to be amorphous.

[0147] None of the salts isolated from this study were recommended for development because the isolation procedure was long, complex, and Compound 1 tended to form gummy substances. However, promising results were obtained suggesting that appropriate salts may be possible with several types of salts, such as zinc and aluminum, and further research is encouraged.

[0148] Study 3 A third salt screening study was conducted. At a 100 mg scale, salt formation reactions were attempted using bases with an approximate pKa greater than 8 or acceptable strong acids with a pKa less than 4. In the first set of salt screening experiments, the counterion sources shown in Table 5 below were examined.

[0149] [Table 5]

[0150] Briefly, 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]methyl-(2,2- diphenylethyl )amino]-butoxy]phenyl]acetic acid was suspended / dissolved in six different solvent systems - isopropanol, acetone, cyclohexanone, methyl ethyl ketone, ethyl acetate, and THF. The corresponding counterion source (1.0 equivalent) was then added as a solution (if possible) or by neat addition. The solution was temperature cycled between two specific temperatures (e.g., 25 °C and 40 °C). All experiments were monitored for precipitation / crystallization. In vials where crystallization / precipitation was not obvious, cooling to below ambient temperature, or cooling to below ambient temperature (e.g., 5 °C), or repeated addition of an appropriate anti-solvent with or without cooling was carried out. In vials where crystallization / precipitation was not obvious after cooling and / or addition of the anti-solvent, the solvent was removed by evaporation. The solid was isolated by filtration and analyzed by XRPD. Oils or gummy substances were mainly obtained.

[0151] A second salt screen was performed with the counterion sources shown in Table 6 below.

[0152]

Table 6

[0153] The procedure for preparing the salt using the counterion source of Table 5 is as follows. According to a known procedure (Collins et al. J. Med. Chem 2002, 45:1963 - 1966), 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]methyl-(2,2- diphenylethyl )amino]butoxy]-phenyl]acetic acid was generated from its hydrochloride salt. 100 mg of the free substance was transferred to a 20 mL vial. Ethyl acetate (1 mL) was added to each vial, followed by the addition of 1.0 equivalent of each counterion source. In a 4-hour cycle, the sample was subjected to a temperature cycle from ambient temperature to 40 °C with stirring for 24 hours. To any clear solution, heptane was added as an anti-solvent, and the sample was placed in the refrigerator for 24 hours. All the solids thus obtained were analyzed by XRPD, and the lids of all the samples that remained clear solutions were removed and evaporated. From these counterion sources, solids identified as the counterion source, or sticky, gummy solids were produced.

[0154] Using an aqueous precipitation approach, other salt screens were performed. In this salt screen, a metathesis procedure was used to prepare the 1:1 sodium salt of Compound 1, and 1.0, 0.5, or 0.3 equivalents of the selected metal counterion were added as its chloride or sulfate salt. Alternatively, the free Compound 1 was reacted with the metal counterion selected as its acetate or sulfate salt. These aqueous precipitations produced free-flowing white solids from zinc, aluminum, and bismuth.

[0155] The counterions shown in Table 7 below were examined.

[0156]

Table 7

[0157] A zinc salt was prepared via metathesis using the following procedure. Compound 1 (200 mg) was dissolved in methanol (2 mL) to form a mixture. Then, 1 M NaOH solution (336 μL, 1.0 equivalent) was added to the mixture to form a 1:1 sodium salt of Compound 1. Zinc chloride (22.5 mg, 0.5 equivalent) was dissolved in water (1 mL), and the resulting solution was added to the mixture, and simultaneously, a solid precipitated from the solution. An additional water (1 mL) was added to the mixture, and the reaction vessel was sealed with paraffin. Thereafter, the mixture was subjected to a temperature cycle at ambient temperature to 40 °C with stirring for 24 h in a 4 h cycle. Methanol was removed from the mixture by rotary evaporation, and water (10 mL) was added to reduce solubility. The resulting precipitate was isolated by Buchner filtration (Whatman grade 1 filter paper, φ = 55 mm) and washed with water (100 mL) to remove residual sodium chloride. The solid was dried on the filter paper to obtain a white powder (152.9 mg, 72%).

[0158] The substance obtained via the above salt metathesis procedure was characterized by ICP, XRPD (Figure 1), PLM (Figure 2), TG / DTA (Figure 3), DSC (Figure 4), 1 H-NMR (Figure 5), and 19 F-NMR (Figure 6) according to the method described in Example 1. From the characterization of the substance, the following information was obtained: · According to ICP analysis, this substance contained 5.27 wt% zinc. The stoichiometric salt is 9.9 wt%, and thus, approximately 0.51 equivalent of zinc (expected value 0.5 equivalent) is present. · According to ICP analysis, this substance contained 0.19 wt% sodium. The stoichiometric salt is 3.7 wt%, and thus, approximately 5% of the substance is the 1:1 sodium salt, and approximately 95% of the substance is the 2:1 zinc salt. · According to XRPD analysis, this substance was amorphous. · This substance showed some birefringence by PLM analysis with an irregular morphology. · There was a minimal mass loss in the TG trace up to decomposition. ·According to DSC, there was an endothermic event at the starting temperature of 51 °C in the first thermal cycle, and there was a sharp event corresponding to glass transition with the midpoint temperature at 52 °C in the second thermal cycle. · 1 The 1H-NMR spectrum was consistent with Compound 1. · 19 19F-NMR showed one main peak. ·The purity of the amorphous substance determined by HPLC analysis is shown in Table 8 below.

[0159]

Table 8

[0160] As shown in Table 8, the zinc salt prepared by the above method is stable after being stored at 40 °C / relative humidity 75% and 80 °C for one week.

[0161] The zinc salt was also prepared using zinc acetate as the counterion source. Briefly, Compound 1 (78 mg) was dissolved in methanol (1.5 mL), and zinc acetate was dissolved in methanol / water (9:1; 1.5 mL). Then, the two solutions thus obtained were combined to form a mixture. A white solid formed and precipitated immediately from the solution. Next, the sample was cycled at ambient temperature to 40 °C for about 5 hours, and then a small amount of white solid remained undissolved. Then, the solvent was removed by rotary evaporation, and methanol (5 mL) was added to the solid. The cycling and solvent evaporation steps were repeated, then methanol / water (9:1; 5 mL) was added to the sample, and the cycling and solvent evaporation steps were repeated once more. Subsequently, the sample was freeze-dried overnight to remove water, and then a white solid was obtained.

[0162] An aluminum salt was prepared via metathesis using the following procedure. Compound 1 (200 mg) was dissolved in methanol (2 mL). A 1 M NaOH stock solution (336 μL, 1.0 equivalent) was added to prepare the sodium salt. Methanol was removed by rotary evaporation. Aluminum sulfate and water (2 mL) were added, and the sample was subjected to temperature cycling at ambient temperature to 40 °C for 24 h in a 4 h cycle. The solid was isolated by centrifugation. Water (3 mL) was added and the slurry was stirred at ambient temperature for 18 h. The solid was filtered using a Büchner funnel (Whatman grade 1 filter paper, φ = 42.5 mm) and washed with water (100 mL) before drying on the filter paper for 10 min. The solid was dried under vacuum at ambient temperature for about 2 h to obtain a white powder (128.7 mg, 61%).

[0163] Example 3 FTIR Analysis Study CHCl 3 A stock solution of the free compound of Compound 1 and the zinc salt of Compound 1 in [CHCl] was prepared. In a volumetric flask, the free form of Compound 1 (50 mg) was diluted to 10 mL with [CHCl]. The zinc salt of Compound 1 (50 mg) was also diluted to 10 mL with [CHCl] in a separate volumetric flask. The FTIR spectrum of the free form of Compound 1 was obtained according to the FTIR method described in Example 1 (see Figure 7), and the spectrum of the zinc salt form of Compound 1 (see Figure 9) was obtained. 3 in 3 in · Free form of Compound 1: 1:1 (w / w) mixture of zinc salt (mixture of free form stock solution (500 μL) and zinc salt stock solution (500 μL)) (see Figure 8); · Free form of Compound 1: 1:4 (w / w) mixture of zinc salt (mixture of free form stock solution (200 μL) and zinc salt stock solution (800 μL)); · Free form of Compound 1: 1:9 (w / w) mixture of zinc salt (mixture of free form stock solution (100 μL) and zinc salt stock solution (900 μL)); · Free form of Compound 1: 5:95 (w / w) mixture of zinc salt (mixture of free form stock solution (50 μL) and zinc salt stock solution (950 μL)); For example, the zinc salt concentration of Compound 1 was increased to obtain the FTIR spectrum of the sample.

[0164] The spectra were compared with each other and with the FTIR spectra of ZnO (Figure 10) and ZnCl 2 (Figure 11).

[0165] The free form of Compound 1 showed a characteristic peak at approximately 1710 cm that was not present in the zinc salt spectrum (see Figure 9). As the percentage of the free form decreased, the peak at 1710 cm was confirmed to shrink. Further, a broad peak at approximately 1590 cm was confirmed when the intensity of the free compound was low, but an increase in intensity was confirmed as the concentration of the zinc salt increased. -1 -1 -1

[0166] Presumably due to the coordination of the carboxyl group of the carboxylic acid with zinc, the zinc salt is evidenced by the disappearance of the IR peak at 1710 cm. Further, the zinc salt does not contain the peaks found in the IR spectrum of ZnO (characterized by a distinct, large, broad band starting at 605 cm and less than 500 cm) or ZnCl -1 -1 -1 2 (characterized by a distinct broad peak at 3386 cm), thus demonstrating that the zinc salt is not merely a mixture of the free compound and inorganic zinc. -1

[0167] Other embodiments Regardless of such documents and similar formats, all documents and similar materials cited in this application, including but not limited to patents, patent applications, articles, books, papers, and web pages, are hereby expressly incorporated by reference in their entirety. If one or more of the incorporated documents and similar materials differ from or conflict with this application, including but not limited to defined terms, term usage, described techniques, etc., this application shall prevail.

[0168] ​​​​​​​Although the method has been described in combination with various embodiments and examples, the method is not limited to such embodiments or examples. On the contrary, as will be understood by those skilled in the art, the present disclosure encompasses various alternatives, modifications, and equivalents.

[0169] The method has been particularly shown and described with reference to specific representative embodiments, but it should be understood that various modifications can be made in form and detail without departing from the spirit and scope of the present disclosure. Accordingly, all embodiments within the scope and spirit of the present disclosure and their equivalents are intended to be claimed. The claims, the description of the methods, systems, and assays of the present disclosure, and the figures should not be read as being limited to the order of the elements described, unless otherwise stated as to their effect.

[0170] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 947,968, filed on December 13, 2019, which is hereby incorporated by reference in its entirety. The present invention includes the following embodiments. (Embodiment 1) The zinc salt of 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]methyl-(2,2-diphenylethyl)amino]butoxy]phenyl]acetic acid (Compound 1). (Embodiment 2) The zinc salt according to Embodiment 1, which is a 2:1 (Compound 1:zinc) salt. (Embodiment 3) The zinc salt according to Embodiment 1 or 2, which is amorphous. (Embodiment 4) The zinc salt according to any one of Embodiments 1 to 3, which has a mass loss of less than 1% until decomposition as measured by thermogravimetric analysis. (Embodiment 5) The aluminum salt of Compound 1. (Embodiment 6) The aluminum salt according to Embodiment 5, which is a 3:1 (Compound 1:aluminum) salt. (Embodiment 7) The aluminum salt according to Embodiment 5 or 6, which is amorphous. (Embodiment 8) The aluminum salt according to any one of Embodiments 5 to 7, which has a mass loss of less than 1% until decomposition as measured by thermogravimetric analysis. (Embodiment 9) A pharmaceutical composition comprising the zinc or aluminum salt according to any one of Embodiments 1 to 8 and a pharmaceutically acceptable excipient. (Embodiment 10) The pharmaceutical composition according to Embodiment 9, which contains less than 1.5% by weight of sodium. (Embodiment 11) The pharmaceutical composition according to Embodiment 10, which substantially does not contain the 1:1 sodium salt of Compound 1. (Embodiment 12) The pharmaceutical composition according to any one of Embodiments 9 to 11, which is in unit dosage form. (Embodiment 13) A method for treating cancer, which comprises administering an effective amount of the zinc or aluminum salt according to any one of Embodiments 1 to 8 or the pharmaceutical composition according to any one of Embodiments 9 to 12. (Embodiment 14) The method according to Embodiment 13, wherein the cancer is breast cancer, colon cancer, renal cell carcinoma, lung cancer, hepatocellular carcinoma, gastric cancer, ovarian cancer, pancreatic cancer, esophageal cancer, prostate cancer, sarcoma, bladder cancer, head and neck cancer, glioblastoma, diffuse large B-cell lymphoma, leukemia, or melanoma. (Embodiment 15) The method according to Embodiment 14, wherein the cancer is metastatic cancer. (Embodiment 16) The method according to Embodiment 15, wherein the effective amount includes an amount effective to suppress the formation of metastatic colonies of the cancer. (Embodiment 17) A method for producing a zinc salt of Compound 1, the method comprising combining Compound 1 or a salt thereof and a zinc salt under conditions sufficient to form the zinc salt of Compound 1. (Embodiment 18) The method according to Embodiment 17, wherein the zinc salt of Compound 1 is a 2:1 (Compound 1:zinc) salt. (Embodiment 19) The method according to Embodiment 17 or 18, wherein the Compound 1 or a salt thereof is a 1:1 (Compound 1:sodium) salt. (Embodiment 20) The method according to any one of Embodiments 17 to 19, wherein the zinc salt is zinc chloride. (Embodiment 21) The method according to any one of Embodiments 17 to 19, wherein the zinc salt is zinc acetate. (Embodiment 22) The method according to any one of Embodiments 17 to 21, comprising dissolving the Compound 1 or a salt thereof and the zinc salt in a solvent to form a mixture. (Embodiment 23) The method according to Embodiment 22, further comprising cycling the temperature of the mixture between ambient temperature and 40 °C. (Embodiment 24) The method according to Embodiment 23, wherein the cycling is performed in a 4-hour cycle. (Embodiment 25) The method according to Embodiment 24, wherein the cycling is performed for 24 hours. (Embodiment 26) A peak having an increase in intensity relative to the free acid at about 1590 ± 10 cm -1 and a peak having a decrease in intensity relative to the free acid at about 1710 ± 10 cm -1 The zinc salt according to Embodiment 3, having the above properties as measured by Fourier transform infrared spectroscopy (FTIR).

Claims

1. The zinc salt of 2-[3-[(3R)-3-[[2-chloro-3-(trifluoromethyl)phenyl]methyl-(2,2-diphenylethyl)amino]butoxy]phenyl]acetic acid (Compound 1), which is a 2:1 (Compound 1:zinc) salt.

2. The zinc salt according to Claim 1, which is amorphous.

3. The zinc salt according to Claim 1 or 2, which has a mass loss of less than 1% until decomposition as measured by thermogravimetric analysis.

4. A pharmaceutical composition comprising the zinc according to any one of Claims 1 to 3 and a pharmaceutically acceptable excipient.

5. The pharmaceutical composition according to Claim 4, wherein the amount of sodium is less than 1.5% by weight.

6. The pharmaceutical composition according to Claim 5, which does not contain the 1:1 sodium salt of Compound 1.

7. The pharmaceutical composition according to any one of Claims 4 to 6, which is in unit dosage form.

8. The pharmaceutical composition according to any one of Claims 4 to 7, for use in a method of treating cancer, which comprises administering the pharmaceutical composition in an effective amount.

9. The pharmaceutical composition according to Claim 8, wherein the cancer is breast cancer, colon cancer, renal cell carcinoma, lung cancer, hepatocellular carcinoma, gastric cancer, ovarian cancer, pancreatic cancer, esophageal cancer, prostate cancer, sarcoma, bladder cancer, head and neck cancer, glioblastoma, diffuse large B-cell lymphoma, leukemia, or melanoma.

10. The pharmaceutical composition according to Claim 9, wherein the cancer is metastatic cancer.

11. The pharmaceutical composition according to Claim 10, wherein the effective amount comprises an amount effective to inhibit metastatic colony formation of the cancer.

12. A peak having an increase in intensity relative to the free acid of Compound 1, measured by Fourier transform infrared spectroscopy (FTIR), at 1590 ± 10 cm -1 and a peak having a decrease in intensity relative to the free acid of Compound 1 at 1710 ± 10 cm -1 The zinc salt according to claim 2, having.

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