OXPHOS INHIBITORS FOR USE IN TREATING CANCER - Patent application

Abstract

The invention features a method of treating cancer in a subject, the method including administering an oxidative phosphorylation (OXPHOS) inhibitor to the subject, where the cancer is refractory to first-line treatment or recurrent.

Description

[Technical field]

[0001] Claiming priority This application claims the benefit of the filing date of U.S. Provisional Patent Application No. 63 / 268,111, filed February 16, 2022, the contents of which are incorporated herein by reference in their entirety.

[0002] Generally, the present invention relates to a method of treating cancer with an inhibitor of oxidative phosphorylation (OXPHOS) in a subject in need thereof. The present invention further relates to treating refractory or recurrent cancer with an inhibitor of oxidative phosphorylation in a subject in need thereof. [Background technology]

[0003] Cancer is a group of diseases caused by harmful, abnormal, uncontrolled and unwanted growth of cells, and appears in multiple tissues and organs, including breast, lung, bladder, colon, rectum, uterus, testes, kidney, blood, lymphatic system, liver, bile duct, skin, pancreas, prostate, thyroid, brain and stomach. Globally, approximately 10,000,000 individuals die from cancer annually. Current therapies for treating cancer may not be able to treat recurrent cancer. Therefore, there is a continuous need for improved therapies for treating cancer. Summary of the Invention

[0004] The present invention provides a method for treating cancer by administering an oxidative phosphorylation (OXPHOS) inhibitor, wherein the cancer is refractory or recurrent. The present invention provides a method for treating cancer by administering a compound of the following formula 1 or a pharma- ceutical acceptable salt thereof: We also provide a method to use TIFF2025506691000001.tif27170.

[0005] In one aspect, the invention features a method of treating cancer in a subject, the method including administering an oxidative phosphorylation (OXPHOS) inhibitor to the subject, where the cancer is refractory to first-line treatment or recurrent.

[0006] In one embodiment, the first-line treatment is a standard of care for cancer. In another embodiment, the cancer is a carcinoma, sarcoma, leukemia or lymphoma. In another embodiment, the cancer is a solid tumor. In an additional embodiment, the cancer is a metastatic cancer.

[0007] In some embodiments, the leukemia is acute lymphocytic leukemia, chronic lymphocytic leukemia, myeloid leukemia, or chronic myelogenous leukemia. In some embodiments, the lymphoma is Hodgkin's lymphoma or non-Hodgkin's lymphoma. In other embodiments, the cancer is breast cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, bladder cancer, colon cancer, rectal cancer, uterine cancer, vaginal or vulvar cancer, testicular cancer, kidney cancer, liver cancer, bile duct cancer, cholangiocarcinoma, gallbladder cancer, melanoma, pancreatic cancer, pancreatic ductal adenocarcinoma, BRAF mutant metastatic melanoma, prostate cancer, castration resistant prostate cancer, thyroid cancer, glioblastoma, endometrial cancer, ovarian cancer, cervical cancer, brain cancer, glioblastoma, liver cancer, colorectal cancer, skin cancer, basal cell carcinoma, squamous cell carcinoma, melanoma, gynecological cancer, head and neck cancer, mesothelioma, myeloma, esophageal cancer, or gastric cancer.

[0008] In certain embodiments, the OXPHOS inhibitor is a compound of formula 1 or a pharma- ceutically acceptable salt thereof.

[0009] In some embodiments, the compound of formula 1 is formulated as a tablet or capsule. In other embodiments, the tablet or capsule is formulated for delayed release. In further embodiments, the tablet or capsule comprises an enteric coating. In some embodiments, the compound of formula 1 is administered at a dose of about 100mg, 200mg, 400mg, 800mg or 1200mg per day. In certain embodiments, the compound of formula 1 is administered at a dose of about 800mg per day.

[0010] In other embodiments, the OXPHOS inhibitor is administered in combination with a second therapeutic compound.

[0011] In another aspect, the invention features a method of treating pancreatic ductal adenocarcinoma (PDAC), the method comprising administering to a subject in need thereof a therapeutically effective dose of a pharmaceutical composition, the composition comprising a compound of Formula 1 or a pharma- ceutical acceptable salt thereof as an active ingredient.

[0012] In one embodiment, the compound of Formula 1 is administered in combination with gemcitabine and abraxane.

[0013] In an additional aspect, the invention features a method of treating BRAF mutant melanoma, the method comprising administering to a subject in need thereof a pharmaceutical composition in a therapeutically effective dose, the composition comprising a compound of Formula 1 or a pharma- ceutical acceptable salt thereof as an active ingredient.

[0014] In some embodiments, the compound of formula 1 is administered in combination with encorafenib and binimetinib. In other embodiments, the BRAF mutant melanoma is metastatic. In some embodiments, the metastasis is to the central nervous system. In certain embodiments, the metastasis is to the brain. In another embodiment of this aspect, the method comprises the use of irradiation and / or the administration of temozolomide.

[0015] In another aspect, the invention features a method of treating castration-resistant prostate cancer (CRPC), the method comprising administering to a subject in need thereof a therapeutically effective dose of a pharmaceutical composition, the composition comprising a compound of Formula 1 or a pharma- ceutical acceptable salt thereof as an active ingredient.

[0016] In some embodiments, the subject has not responded to at least one course of immunotherapy. In other embodiments, the compound of formula 1 is administered in combination with a checkpoint inhibitor. In certain embodiments, the checkpoint inhibitor is a PD-1 inhibitor.

[0017] In some embodiments, the pharma- ceutically acceptable salt is a salt of an acid selected from the group consisting of formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, trifluoroacetic acid, malic acid, maleic acid, malonic acid, fumaric acid, succinic acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, benzensulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, dichloroacetic acid, aminooxyacetic acid, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, and boric acid.

[0018] In certain embodiments, the pharma- ceutically acceptable salt is a salt of acetic acid.

[0019] In some embodiments, the subject is a human.

[0020] definition In this application, unless otherwise clear from the context, (i) the term "a" may be understood to mean "at least one;" (ii) the terms "comprising" and "including" may be understood to encompass the listed components or steps, whether presented alone or in combination with one or more additional components or steps; (iii) the term "approximately" may be understood to allow for standard deviation as would be understood by one of ordinary skill in the art; and (iv) when a range is provided, the endpoints are included.

[0021] As used herein, the term "about" refers to a value that is within ±10% of the value that follows the term "about". Reference herein to "about" a value or parameter includes (and describes) the variation directed to the value or parameter itself. For example, a statement that refers to "about X" includes the statement of "X".

[0022] As used herein, the term "administration" refers to the administration of a composition (e.g., a compound or preparation comprising an OXPHOS inhibitor) to a subject or system. Administration to an animal subject (e.g., to a human) may be by any suitable route. For example, in some embodiments, administration may be bronchial (including by bronchial instillation), buccal, enteral, interdermal, intraarterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, intravaginal and intravitreal.

[0023] The "effective amount" of a compound (e.g., an OXPHOS inhibitor) may vary according to factors such as the individual's condition, age, sex, and weight, and the ability of the compound to induce a desired response. A therapeutically effective amount includes an amount in which any toxic or harmful effects of the compound are outweighed by the therapeutically beneficial effects. An effective amount also includes an amount sufficient to provide a benefit, such as a clinical benefit.

[0024] As used herein, the term "subject" or "participant" or "patient" refers to any organism to which a compound or composition according to the present invention may be administered, for example, for experimental, diagnostic, prophylactic and / or therapeutic purposes. Exemplary subjects include any animal (e.g., mammals such as dogs, mice, rats, rabbits, pigs, non-human primates and humans). A subject may be a human or animal that is seeking or in need of treatment, requesting treatment, undergoing treatment, or about to undergo treatment, or receiving the care of a trained professional for a particular disease or condition.

[0025] As used herein, the terms "treat", "treated" or "treating" refer to both therapeutic treatment and prophylactic or preventative measures, where the purpose is to prevent or slow down (alleviate) an undesirable physiological condition, disorder or disease, or to obtain a beneficial or desired clinical outcome. Beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms; reduction in the extent of a condition, disorder or disease; a stable (i.e., non-worsening) status of a condition, disorder or disease; delay in the onset of, or slowing down the progression of, a condition, disorder or disease; improvement or remission (whether partial or total) of a condition, disorder or pathology, whether detectable or not; an improvement in at least one measurable physical parameter, not necessarily discernible by the subject; or enhancement or amelioration of a condition, disorder or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival compared to expected survival in the absence of treatment.

[0026] As used herein, the term "checkpoint inhibitor" refers to an inhibitor of a protein, i.e., a checkpoint protein, that determines whether immune cells target cells for destruction. Cancer cells express proteins on their surface that bind to checkpoint proteins on T cells. The binding between a ligand on the surface of a cancer cell and a checkpoint inhibitor on an immune cell cannot cause the immune cell to destroy the cancer cell. A checkpoint inhibitor blocks the interaction between a ligand on the surface of a cancer cell and a checkpoint inhibitor, causing the immune cell to destroy the ligand-bearing cancer cell.

[0027] As used herein, the term "PD-1 inhibitor" refers to an inhibitor of programmed death ligand-1 (PD1), a checkpoint inhibitor expressed on the surface of T cells and B cells. Exemplary and non-limiting PD-1 inhibitors include pembrolizumab, nivolumab, cemplimab, and dostallimab.

[0028] As used herein, the term "recurrent cancer" refers to the growth of cancer cells that appears in a subject after the subject has undergone treatment for cancer.Cancer recurrence can be local recurrence if the cancer reappears in the same place where it was originally found, regional recurrence if the cancer reappears in lymph nodes near where it was originally found, or distant reoccurrence if the cancer reappears in a different part of the body than where it was originally found.

[0029] As used herein, the term "refractory cancer" is a cancer that does not respond to first line treatment, such as standard therapy, or that recurs after an initial response.

[0030] As used herein, the term "standard of care" means the treatment that a reasonably prudent physician would administer to a subject suffering from a disease or disorder.

[0031] As used herein, the term "carcinoma" refers to cancer that begins in the skin or in the tissues that surround or cover internal organs.

[0032] As used herein, the term "lymphoma" refers to a cancer that begins in the cells of the immune system.

[0033] As used herein, the term "leukemia" refers to cancer that begins in the blood cells or blood-forming tissues.

[0034] As used herein, the term "sarcoma" refers to cancer that begins in the bone or in the soft tissues of the body, including cartilage, fat, muscle, blood vessels, fibrous tissue, connective tissue, or supportive tissue.

[0035] As used herein, an "enteric coating" is a polymeric material or materials that encases the pharmaceutical core (e.g., containing N1-pyrrolidine-N5-(3-trifluoromethoxy)phenyl biguanide (a compound of formula 1) or a pharma- ceutically acceptable salt thereof) as it passes through the stomach. Suitable pH-sensitive polymers are those that dissolve in intestinal fluids at higher pH levels (pH greater than 4.5), such as in the small intestine, thereby allowing release of the pharmacologically active agent in the region of the small intestine, but not in the upper part of the GI tract, such as the stomach. The enteric coating also protects the stomach from irritating pharmaceutical ingredients. The enteric coating may be applied to a variety of formulations, including tablets, capsules, and microparticles. The enteric coating may be applied directly to the pharmaceutical core or may be an integral part of the tablet, capsule, or microparticle. Alternatively, the enteric coating may be separate from the pharmaceutical core, in which case the coating is not attached to the pharmaceutical core while encapsulating it.

[0036] As used herein, "delayed release" refers to a pharmaceutical preparation, such as an orally administered formulation, that passes through the acidic environment of the stomach substantially intact and dissolves in the more basic environment of the small intestine.In some embodiments, the delayed release of an active agent (e.g., N1-pyrrolidine-N5-(3-trifluoromethoxy)phenyl biguanide (the compound of formula 1) or its pharma-ceutically acceptable salt) occurs by using a pH-sensitive enteric coating of an oral dosage form.The enteric coating may be combined, for example, with a delayed release formulation, to extend the time it takes for the drug to be released.

[0037] As used herein, an "effective amount" means an amount of an agent sufficient to effect a beneficial or desired result in a patient, such as disease remission, and therefore an "effective amount" will depend on the context in which it is applied, including the age and weight of the patient, the organ affected by the disease, the nature of the disease, including pathology or level of activity, and other factors.

[0038] As used herein, "pharmaceutically acceptable salt" refers to a salt that is suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic response, etc., within the scope of sound medical judgment, and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and in Pharmaceutical Salts: Properties, Selection, and Use (eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. Salts can be prepared in situ during the final isolation and purification of the compounds of the present invention, or separately by reacting the free base group with a suitable organic or inorganic acid. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, bitartrate, borate, butyrate, camphor, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, and 2-hydroxy-ethanesulfonate. Salts include, but are not limited to, glyceryl stearate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate, and the like.Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc., as well as non-toxic ammonium, quaternary ammonium and amine cations, including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, etc. In one embodiment, the pharma- ceutically acceptable salt is a salt of acetic acid.

[0039] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs.Methods and materials for use in this disclosure are described herein, and other suitable methods and materials known in the art can also be used.Materials, methods and examples are illustrative only and are not intended to be limiting.All publications, patent applications, patents, sequences, database entries and other references mentioned herein are incorporated herein in their entirety by reference.In case of conflict, the present specification, including definitions, will prevail.

[0040] The details of one or more embodiments of the invention are set forth in the drawings and the description which follow. Other features, objects, and advantages of the invention will be apparent from the description, and from the claims. [Brief description of the drawings]

[0041] [Figure 1A] FIG. 1 is a plot showing pre- and post-treatment biopsies for 84 unique cancer therapies. [Figure 1B] FIG. 1 is a plot showing an increase in OXPHOS in post-treatment biopsies compared to pre-treatment biopsies and no corresponding increase in glycolysis in post-treatment biopsies compared to pre-treatment biopsies. [Figure 1C] FIG. 1 is a plot showing increased OXPHOS following leuprolide treatment in breast and prostate cancer. [Figure 1D]FIG. 1 is a plot showing increased OXPHOS following enzalutamide treatment in prostate cancer. [Figure 1E] FIG. 1 is a plot showing increased OXPHOS following fulvestrant, letrozole and palbociclib treatment in breast cancer. [Figure 1F] FIG. 1 is a plot showing increased OXPHOS following cyclophosphamide treatment in breast cancer and sarcoma. [Figure 1G] FIG. 1 is a plot showing increased OXPHOS following carboplatin treatment in breast, endometrial, non-small cell lung cancer (NSCLC) and ovarian cancer. [Figure 1H] FIG. 1 is a plot showing increased OXPHOS following doxorubicin treatment in bladder cancer, breast cancer and sarcoma. [Figure 1I] FIG. 1 is a plot showing increased OXPHOS following paclitaxel treatment in breast, endometrial, NSCLC and ovarian cancer. [Figure 1J] FIG. 1 is a plot showing increased OXPHOS following temozolomide treatment in glioblastoma. [Diagram 2] FIG. 1 is a plot showing that in vitro exposure of patient-derived xenograft cells to gemcitabine sensitizes tumor cells to the compound of Formula 1. [Diagram 3] 1 is a graph showing that a compound of Formula 1 in combination with gemcitabine increases animal survival. [Figure 4A] 1 is a photomicrograph showing expression of pAMPK in cells untreated or treated with a compound of Formula 1. [Figure 4B] 1 is a photomicrograph showing expression of HIF-1a in cells untreated or treated with a compound of Formula 1. [Diagram 5] FIG. 1 describes a Phase 2 clinical study protocol evaluating the use of the compound of Formula 1 in combination with standard of care (SOC) for the treatment of BRAF mutant metastatic melanoma. [Figure 6]FIG. 1 describes a Phase 1b / 2a clinical study protocol evaluating the use of the compound of Formula 1 in combination with standard of care (SOC) for the treatment of pancreatic ductal adenocarcinoma. [Figure 7A] 1 is a graph showing the response of cell subtypes from glioblastoma patients to compounds of Formula 1. [Figure 7B] 1 is a chart showing the differential response of MTC and GPM cell subtypes to compounds of Formula 1. [Figure 8] FIG. 1 describes a Phase 2 clinical study protocol evaluating the use of the compound of Formula 1 in the treatment of patients with glioblastoma or BRAF mutant melanoma and brain metastes. [Figure 9] FIG. 1 describes a Phase 2 clinical study protocol evaluating the use of the compound of Formula 1 in combination with standard of care (SOC) for the treatment of castration-resistant prostate cancer. [Figure 10] 1 is a chart describing the therapeutic window of compounds of formula 1. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0042] The present inventors have discovered that cancer cells with high oxidative phosphorylation (OXPHOS) levels can survive treatment with therapeutic agents that target metabolic pathways other than OXPHOS, and can recur in subjects after treatment with therapeutic agents that do not target OXPHOS. Thus, the present invention provides a method of treating cancer with an OXPHOS inhibitor in a subject in need thereof.

[0043] OXPHOS inhibitors OXPHOS is a metabolic pathway that involves the oxidation of chemical bonds in molecules, such as glucose, resulting in the production of adenosine triphosphate (ATP).The method of the present invention encompasses all inhibitors of OXPHOS, such as substances that inhibit proteins in OXPHOS pathways, such as enzymes that catalyze chemical reactions in OXPHOS metabolic pathways, and substances that inhibit the expression of proteins in OXPHOS metabolic pathways.

[0044] Exemplary, non-limiting OXPHOS inhibitors that may be used according to the methods of the invention include biguanide compounds, such as metformin, phenformin or buformin, BAY84-243, carboxyamidotriazole (CAI), fenofibrate, meta-iodobenzylguanidine (mIBG), pyrvinium, canagliflozin, pioglitazone, rosiglitazone, amobarbital, nefazodone, alphaTOS, lonidamine, atovaquone, arsenic trioxide, nitric oxide, hydrocortisone, or uncoupling proteins (UCPs), such as UCP1, UCP2, UCP3, UCP4 or UCP5, or derivatives or conjugates thereof.

[0045] In some embodiments, the OXPHOS inhibitor is N-1-pyrrolidine-N-5-(3-trifluoromethoxy)phenyl biguanide, a compound of formula 1, or a pharma- ceutically acceptable salt thereof. TIFF2025506691000002.tif27170

[0046] Methods for synthesizing compounds of formula 1 are disclosed in US Pat. No. 9,540,325, which is incorporated by reference in its entirety into this application.

[0047] Any suitable OXPHOS inhibitor, e.g., an OXPHOS inhibitor with an ionizable functional group, may be administered as a pharma- ceutically acceptable salt, e.g., a salt of an acid selected from the group consisting of formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, trifluoroacetic acid, malic acid, maleic acid, malonic acid, fumaric acid, succinic acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, dichloroacetic acid, aminooxyacetic acid, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, and boric acid.

[0048] Treatment OXPHOS inhibitors can be used to treat cancer. Cancer is a group of diseases characterized by harmful, abnormal, uncontrolled and unwanted growth of cells. In some embodiments, the uncontrolled growth is due to cells dividing and growing in the absence of signals (e.g., growth factors) that instruct them to do so. In some embodiments, the uncontrolled growth is due to cells that cannot respond to signals that instruct cells to stop growing and / or to engage in programmed cell death (i.e., apoptosis). In some embodiments, cancer cells may spread (i.e., metastasize) throughout the body. In some embodiments, cancer cells may form tumors. In some embodiments, cancer cells may form solid tumors.

[0049] The following description of the treatment of cancer with OXPHOS inhibitors is provided without wishing to be bound by theory. In some embodiments, cancer cells have metabolic activities, such as Krebs cycle and oxidative phosphorylation (OXPHOS) activities, that are unique to non-cancer cells' metabolic activities, such as Krebs cycle and OXPHOS activities. In some embodiments, OXPHOS inhibitors target the unique metabolic activities of cancer cells. In some embodiments, targeting the unique metabolic activities of cancer cells allows OXPHOS inhibitors to selectively kill cancer cells over non-cancer cells.

[0050] In some embodiments, the total population of cancer cells, for example the total population of cancer cells in a subject, the total population of cancer cells in an organ, the total population of cancer cells in a biological tissue, or the total population of cancer cells in a sample of cancer cells, may be composed of multiple subpopulations of cancer cells with distinct metabolic activities. One subpopulation of cancer cells may rely on OXPHOS to meet their metabolic needs, while a second subpopulation of cancer cells may not rely on OXPHOS to meet their metabolic needs, but may instead rely on an alternative metabolic pathway, for example glycolysis.

[0051] In some embodiments, treatment of a population of cancer cells, for example a population of cancer cells in a subject, with a therapeutic agent that is not an OXPHOS inhibitor can target only cancer cells that do not rely on OXPHOS, and can result in the death of only cancer cells that do not rely on OXPHOS. In some embodiments, treatment of a population of cancer cells with a therapeutic agent that targets a metabolic pathway other than OXPHOS can lead to the growth of cancer cells that rely on OXPHOS, for example the reappearance of OXPHOS-dependent cancer cells in a subject.

[0052] In some embodiments, treatment of a population of cancer cells with both non-OXPHOS-targeting therapeutics and OXPHOS-targeting therapeutics, i.e., OXPHOS inhibitors, may result in the killing of both OXPHOS-dependent and non-OXPHOS-dependent cancer cells.

[0053] In some embodiments, the non-OXPHOS targeting therapeutic agent and the OXPHOS targeting therapeutic agent may be administered sequentially. In some embodiments, the non-OXPHOS targeting therapeutic agent and the OXPHOS targeting therapeutic agent may be administered simultaneously.

[0054] In some embodiments, OXPHOS inhibitors can be used to treat the re-emergence of cancer cells, for example, the re-emergence of OXPHOS-dependent cancer cells that survive treatment with non-OXPHOS-targeting therapeutic agents.In some embodiments, the simultaneous treatment of a population of cancer cells with OXPHOS-targeting therapeutic agents and non-OXPHOS-targeting therapeutic agents results in the death of both OXPHOS-dependent cancer cells and non-OXPHOS-dependent cancer cells.In some embodiments, the simultaneous treatment of a population of cancer cells with OXPHOS-targeting therapeutic agents and non-OXPHOS-targeting therapeutic agents prevents the re-emergence of cancer cells, for example, the re-emergence of OXPHOS-dependent cancer cells.

[0055] Thus, the OXPHOS inhibitor can be administered to the subject in an effective amount to treat cancer.Non-limiting examples of cancer that can be treated by the OXPHOS inhibitor or its pharmaceutically acceptable salt include carcinoma, sarcoma, leukemia (e.g., acute lymphocytic leukemia, chronic lymphocytic leukemia, myelogenous leukemia or chronic myelogenous leukemia) and lymphoma (e.g., Hodgkin's lymphoma or non-Hodgkin's lymphoma).In some embodiments, the cancer is a solid tumor.In some embodiments, the cancer is metastasis. Particular types of cancer that may be treated by an OXPHOS inhibitor or a pharma- ceutically acceptable salt thereof include breast cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, bladder cancer, colon cancer, rectal cancer, uterine cancer, vaginal or vulvar cancer, testicular cancer, kidney cancer, liver cancer, bile duct cancer, cholangiocarcinoma, gallbladder cancer, melanoma, pancreatic cancer, pancreatic ductal adenocarcinoma, BRAF mutant metastatic melanoma, prostate cancer, castration-resistant prostate cancer, thyroid cancer, glioblastoma, endometrial cancer, ovarian cancer, cervical cancer, brain cancer, glioblastoma, liver cancer, colorectal cancer, skin cancer, basal cell carcinoma, squamous cell carcinoma, melanoma, gynecological cancer, head and neck cancer, mesothelioma, myeloma, esophageal cancer, and gastric cancer.

[0056] Pancreatic ductal adenocarcinoma Pancreatic ductal adenocarcinoma (PDAC) is an aggressive form of pancreatic cancer that arises in the lining of the pancreatic ducts. PDAC is the most common form of pancreatic cancer and accounts for more than 90% of pancreatic malignancies worldwide. PDAC is the fourth leading cause of cancer-related deaths worldwide and has an overall 5-year survival rate of less than 8%.

[0057] BRAF-mutated metastatic melanoma Metastatic melanoma is a form of melanoma that has spread to a location in the body other than where the melanoma began. BRAF-mutated metastatic melanoma is a form of metastatic melanoma in which the gene that codes for the B-Raf protein, the BRAF gene, contains one or more mutations. BRAF mutations are the most commonly found mutations in metastatic melanoma. Metastatic melanomas with BRAF mutations can be more aggressive than those without BRAF mutations. BRAF-mutated metastatic melanomas can metastasize to the brain.

[0058] glioblastoma Glioblastoma (GBM) is an aggressive type of cancer that can occur in the brain or spinal cord and arises from glial cells. The overall five-year survival rate for subjects diagnosed with GBM and receiving standard treatment is less than 7%, and the median survival time for subjects diagnosed with GBM is estimated to be 12 to 18 months.

[0059] Castration-resistant prostate cancer Castration-resistant prostate cancer is a form of prostate cancer that continues to grow even when the amount of testosterone in a subject is reduced to very low levels, for example after surgical or medical castration. The estimated average survival of subjects diagnosed with castration-resistant prostate cancer is approximately 9 to 36 months, and subjects diagnosed with castration-resistant prostate cancer and undergoing standard treatment have a poor quality of life.

[0060] Compositions for Combination Therapy The OXPHOS inhibitor or a pharma- ceutically acceptable salt thereof may be administered with or without a second active ingredient. Non-limiting examples of second active ingredients for treating cancer that may be administered with the OXPHOS inhibitor include lapatinib (Tykerb®), temozolomide (Temodar®), irinotecan (Camptosar®), olaparib (Lynparza®), doxorubicin (Adriamycin®), temsirolimus (Torisel®), gemcitabine (Gemzar®), erlotinib (Tarceva®), carboplatin (Parser®), and rivaroxaban (Ricin®). aplatin®), paclitaxel (Taxol®), bevacizumab (Avastin®), ipilimumab (Yervoy®), gefitinib (Iressa®), apatinib, vandetanib (Caprelsa®), sorafenib (Nexavar®), dabrafenib (Tafinlar®), vemurafenib (Zelboraf®), fulvestrant (Faslodex®), Alpelisib (Piqray®), palbociclib (Ibrance®), letrozole (Femara®), trastuzumab (Herceptin®), pertuzumab (Perjeta®), nelfinavir (Viracept®), bortezomib (Velcade®), disulfiram (Antabuse®), ritonavir (Norvir®), memantine hydrochloride (Namenda®), mefloquine (Lari am®), chloroquine, metronomic cyclophosphamide, liposomal doxorubicin (Caelyx®), docetaxel (Taxotere), rituximab (Rituxan®), cyclophosphamide, vincristine (Marqibo®), prednisone (Rayos®), everolimus (Zortress®), exemestane (Aromasin®), temsiroliumus (Torisel®),These include sapanisertib, rapamycin (Rapamune®), trametinib (Mekinist®), sintilimab (Tyvyt®), nivolumab (Opdivo®), zurvalumab (Imfinzi®), pembrolizumab (Keytruda®) or rosiglitazone (Avandia®).

[0061] Pharmaceutical Compositions The administration of the OXPHOS inhibitor may be by any suitable means that results in the treatment of cancer. The OXPHOS inhibitor may be contained in any suitable amount in any suitable carrier substance, and is generally present in an amount of 1-95% by weight of the total weight of the composition. The composition may be provided in a dosage form that is suitable for sublingual, buccal, oral, parenteral (e.g., intravenously, intramuscularly), pulmonary, intranasal, transdermal, vaginal or rectal administration routes. Thus, the composition may be in the form of, for example, a tablet, capsule, pill, powder, granule, suspension, emulsion, solution, gel, including hydrogel, paste, ointment, cream, plaster, drench, osmotic delivery device, suppository, enema, injection, spray, vapor or aerosol. Pharmaceutical compositions may be formulated according to conventional pharmaceutical practice (see, e.g., Remington: The Science and Practice of Pharmacy (23rd ed.) A. Adejare., ed., 2020, Academic Press, Philadelphia, PA).

[0062] The pharmaceutical compositions according to the present invention may be formulated to release the active compound substantially immediately after administration or at any predetermined time or period after administration. The latter type of composition is generally known as a controlled release formulation, which includes (i) a formulation that provides a substantially constant concentration of the active compound in the body over an extended period of time; (ii) a formulation that provides a substantially constant concentration of the active compound in the body over an extended period of time after a predetermined lag time; and (iii) a formulation that sustains the action of the active compound over a predetermined period of time by maintaining a relatively constant effective level of the active compound in the body while minimizing the undesirable side effects associated with fluctuations in the plasma level of the active compound (sawtooth kinetic pattern).

[0063] In order to obtain the controlled release of the active compound of interest, any one of a number of strategies can be pursued.In one example, controlled release is obtained by appropriate selection of various formulation parameters and raw materials, including, for example, various types of controlled release compositions and coatings.Therefore, drugs are formulated with suitable additives into pharmaceutical compositions that release active compounds in a controlled manner when administered.Examples include single or multiple unit tablet or capsule compositions, oily solutions, suspensions, emulsions, microcapsules, microspheres, nanoparticles, patches, and liposomes.

[0064] The pharmaceutical compositions contemplated by the present invention may be an OXPHOS inhibitor or a pharma- ceutically acceptable salt thereof in admixture with non-toxic pharma- ceutically acceptable additives. Pharmaceutically acceptable additives are known to those skilled in the art. Additives include, for example, sucrose, sorbitol, sugars, mannitol, microcrystalline cellulose, in particular microcrystalline cellulose PH101 or microcrystalline cellulose PH200, starch, including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate or sodium phosphate, or other inert diluents or fillers; crospovidone, sodium alginate, colloidal magnesium, aluminum silicate, calcium silicate, sodium starch glycolate, acrylic acid derivatives, microcrystalline cellulose, sodium carboxymethylcellulose, calcium carboxymethylcellulose, modified cellulose gum, crosslinking agents, such as cellulose acetate, cellulose acetate esters ... Disintegrants such as bridge povidone, alginic acid and alginates, pregelatinized starch, modified corn starch cellulose derivatives including microcrystalline cellulose, starches including potato starch, croscarmellose sodium, alginates or alginic acid; binders such as sucrose, glucose, sorbitol, acacia, alginic acid, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, hydroxypropyl cellulose EXF, ethylcellulose, polyvinylpyrrolidone or polyethylene glycol;Colloidal silicon dioxide, in particular colloidal silicon dioxide Cab-O-Sil® M5P, glycerol tribehenate, magnesium stearate, calcium stearate, talc, sodium stearyl fumarate, sodium behenate, stearic acid, cetyl alcohol, polyoxyethylene glycol, leucine, sodium benzoate, stearates, polyethylene glycol, glyceryl monostearate, glyceryl palmitostearate, liquid paraffin, poloxamer, sodium lauryl sulfate, magnesium lauryl sulfate, hydrogenated castor colloidal silicon dioxide. lubricants and / or glidants such as calcium carbonate, palmitostearate, stearic acid, zinc stearate, stearyl alcohol, silica or hydrogenated vegetable oils; anti-caking agents such as colloidal silicon dioxide, microcrystalline cellulose, tricalcium phosphate, microcrystalline cellulose, magnesium stearate, sodium bicarbonate, sodium ferrocyanide, potassium ferrocyanide, calcium ferrocyanide, calcium phosphate, sodium silicate, colloidal silicon dioxide, calcium silicate, magnesium trisilicate, talcum powder, sodium aluminosilicate, potassium aluminosilicate, calcium aluminosilicate, bentonite, aluminium silicate, stearic acid, polydimethylsiloxane; other pharmaceutically acceptable excipients may be colouring agents, flavouring agents, plasticisers, humectants and buffering agents;

[0065] Dosage The dosage of the OXPHOS inhibitor used in the methods described herein, for example, the OXPHOS inhibitor having the structure of formula I, may vary depending on many factors, such as the age, health and weight of the recipient; the nature and severity of symptoms; the frequency of treatment, and the type of co-treatment, if any; and the clearance rate of the compound in the treated animal. Those skilled in the art can determine the appropriate dosage based on the above factors. The composition used in the methods described herein may be initially administered at a suitable dosage, which may be adjusted as needed according to clinical response. In general, the suitable daily dosage of the compound of the present invention will be the amount of the compound that is the lowest dosage effective to produce a therapeutic effect. Such an effective dosage will generally depend on the factors mentioned above.

[0066] The effective amount of the composition of the present invention may be determined by the subject's body weight, although the attending physician will ultimately decide the appropriate amount and dosage regimen. For example, an effective amount may be between about 8 mg once a day (QD) and about 6,400 mg QD, e.g., between about 8 mg QD and about 3,200 mg QD, between about 8 mg QD and about 3,200 mg QD, between about 8 mg QD and about 1,200 mg QD, between about 8 mg QD and about 800 mg QD, e.g., between about 16 mg QD and about 400 mg QD, between about 32 mg QD and about 200 mg QD, between about 64 mg QD and 100 mg QD, between about 100 mg QD and 400 mg QD, between about 200 mg QD and about 300 mg QD, between about 400 mg QD and about 1,200 mg QD, between about 600 mg QD and about 1,000 mg QD, between about 700 mg QD and about 900 mg QD, between about 750 mg QD and about 850 mg QD, Between the QDs may be a solid dosage form or composition described herein. In some embodiments, the effective amount is between 8 mg QD±0.8 mg QD to 6,400 mg QD±640 mg QD, 8 mg QD±0.8 mg QD to 3,200 mg QD±320 mg QD, 8 mg QD±0.8 mg QD to 2,400 mg QD±240 mg QD, 8 mg QD±0.8 mg QD to 1,200 mg QD±160 mg QD, 8 mg QD±0.8 mg QD to 800 mg QD±80 mg QD, between 16 mg QD±1.6 mg QD to 400 mg QD±40 mg QD, between 32 mg QD±3.2 mg QD to 200 mg QD±20 mg QD, between 64 mg QD±6.4 mg QD to 100 mg QD±10 mg QD, between 100 mg QD±10 mg QD to 400 mg QD The solid dosage form or composition may be between 200 mg QD ± 20 mg QD and 300 mg QD ± 30 mg QD, between 400 mg QD ± 40 mg and 1,200 mg QD ± 120 mg, between 600 mg QD ± 60 mg and 1,000 mg QD ± 100 mg, between 700 mg QD ± 70 mg and 900 mg QD ± 90 mg, between 750 mg QD ± 75 mg and 850 mg QD ± 80 mg.In some embodiments, an effective amount may be between 8 mg QD and 6,400 mg QD, between 8 mg QD and 3,200 mg QD, between 8 mg QD and 2,400 mg QD, between 8 mg QD and 1,200 mg QD, between 8 mg QD and 800 mg QD, between 16 mg QD and 400 mg QD, between 32 mg QD and 200 mg QD, between 64 mg QD and 100 mg QD, between 100 mg QD and 400 mg QD, between 200 mg QD and 300 mg QD, between 400 mg QD and 1,200 mg QD, between 600 mg QD and 1,000 mg QD, between 700 mg QD and 900 mg QD, between 750 mg QD and 850 mg QD of a solid dosage form or composition described herein.

[0067] In some embodiments, this may be a dose of about 8 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of about 16 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of about 32 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of about 64 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of about 100 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of about 200 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of about 300 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of about 400 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of about 800 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be about 1,200 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be about 2,400 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be about 3,200 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be about 6,400 mg QD of the solid dosage form or composition described herein.

[0068] In some embodiments, this may be a dose of 8 mg QD ± 0.8 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of 16 mg QD ± 1.6 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of 32 mg QD ± 3.2 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of 64 mg QD ± 6.4 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of 100 mg QD ± 10 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of 200 mg QD ± 20 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of 300 mg QD ± 30 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of 400 mg QD ± 40 mg QD of the solid dosage forms or compositions described herein. In some embodiments, this may be a dose of 800 mg QD ± 80 mg QD of the solid dosage forms or compositions described herein. In some embodiments, this may be a daily dose of 1,200 mg QD ± 120 mg of the solid dosage forms or compositions described herein. In some embodiments, this may be a daily dose of 2,400 mg QD ± 240 mg of the solid dosage forms or compositions described herein. In some embodiments, this may be a daily dose of 3,200 mg ± 320 mg of the solid dosage forms or compositions described herein. In some embodiments, this may be a daily dose of 6,400 mg ± 640 mg of the solid dosage forms or compositions described herein.

[0069] In some embodiments, this may be a dose of 8 mg QD of a solid dosage form or composition described herein. In some embodiments, this may be a dose of 16 mg QD of a solid dosage form or composition described herein. In some embodiments, this may be a dose of 32 mg QD of a solid dosage form or composition described herein. In some embodiments, this may be a dose of 64 mg QD of a solid dosage form or composition described herein. In some embodiments, this may be a dose of 100 mg QD of a solid dosage form or composition described herein. In some embodiments, this may be a dose of 200 mg QD of a solid dosage form or composition described herein. In some embodiments, this may be a dose of 300 mg QD of a solid dosage form or composition described herein. In some embodiments, this may be a dose of 400 mg QD of a solid dosage form or composition described herein. In some embodiments, this may be a dose of 800 mg QD of a solid dosage form or composition described herein. In some embodiments, this may be a dose of 1,200 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of 2400 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of 3200 mg QD of the solid dosage form or composition described herein. In some embodiments, this may be a dose of 6400 mg QD of the solid dosage form or composition described herein.

[0070] In some embodiments, this may be about 1 mg / kg to about 80 mg / kg of the daily dosage of the solid dosage forms or compositions described herein. In some embodiments, this may be about 1 mg / kg to about 40 mg / kg of the daily dosage of the solid dosage forms or compositions described herein. In some embodiments, this may be about 3 mg / kg to about 30 mg / kg of the daily dosage of the solid dosage forms or compositions described herein. In some embodiments, this may be about 5 mg / kg to about 25 mg / kg of the daily dosage of the solid dosage forms or compositions described herein. In some embodiments, this may be about 10 mg / kg to about 20 mg / kg of the daily dosage of the solid dosage forms or compositions described herein. In some embodiments, this may be about 12 mg / kg to about 18 mg / kg of the daily dosage of the solid dosage forms or compositions described herein. In some embodiments, this may be about 1.25 mg / kg to about 15 mg / kg of the daily dosage of the solid dosage forms or compositions described herein. In some embodiments, this may be about 2.5 mg / kg to about 10 mg / kg of the daily dosage of the solid dosage forms or compositions described herein. In some embodiments, this may be about 5 mg / kg to about 8 mg / kg of the daily dosage of the solid dosage forms or compositions described herein. In some embodiments, this may be 1 mg / kg ± 0.1 mg / kg to 80 mg / kg ± 8300.0 mg / kg of the daily dosage of the solid dosage forms or compositions described herein. In some embodiments, this may be 1 mg / kg ± 0.1 mg / kg to 40 mg / kg ± 4.0 mg / kg of the daily dosage of the solid dosage forms or compositions described herein. In some embodiments, this may be a daily dosage of 3 mg / kg ± 0.3 mg / kg of a solid dosage form or composition described herein to 30 mg / kg ± 3 mg / kg of a solid dosage form or composition described herein.In some embodiments, this may be a daily dose of 5 mg / kg ± 0.5 mg / kg to about 25 ± 2.5 mg / kg of the solid dosage forms or compositions described herein. In some embodiments, this may be a daily dose of 10 mg / kg ± 1.0 mg / kg to about 20 ± 2 mg / kg of the solid dosage forms or compositions described herein. In some embodiments, this may be a daily dose of 12 mg / kg ± 1.2 mg / kg to 18 mg / kg ± 1.8 mg / kg of the solid dosage forms or compositions described herein. In some embodiments, this may be a daily dose of 1.25 ± 0.125 mg / kg to 15 ± 1.5 mg / kg of the solid dosage forms or compositions described herein. In some embodiments, this may be a daily dose of 2.5 mg / kg ± 0.25 to 10 ± 1.0 mg / kg of the solid dosage forms or compositions described herein. In some embodiments, this may be 5±0.5 mg / kg to 8±0.8 mg / kg of the solid dosage forms or compositions described herein. In some embodiments, this may be 1 mg / kg to 40 mg / kg of the solid dosage forms or compositions described herein. In some embodiments, this may be 3 mg / kg to 30 mg / kg of the solid dosage forms or compositions described herein. In some embodiments, this may be 5 mg / kg to 25 mg / kg of the solid dosage forms or compositions described herein. In some embodiments, this may be 10 mg / kg to 20 mg / kg of the solid dosage forms or compositions described herein. In some embodiments, this may be 12 mg / kg to 18 mg / kg of the solid dosage forms or compositions described herein. In some embodiments, this may be 1.25 mg / kg to 15 mg / kg of the solid dosage forms or compositions described herein. In some embodiments, this may be a daily dosage of 2.5 mg / kg to 10 mg / kg of a solid dosage form or composition described herein.In some embodiments, this may be a daily dose of 5mg / kg to 8mg / kg of the solid dosage form or composition described herein. In some embodiments, this may be a daily dose of about 30mg / kg. In some embodiments, this may be a daily dose of about 50mg / kg. In some embodiments, this may be a daily dose of about 60mg / kg.

[0071] Release Profile Controlled release compositions for oral use may be constructed to release the active drug by controlling the dissolution and / or diffusion of the active drug substance. Any of a number of strategies can be pursued to obtain controlled release and a target plasma concentration versus time profile. In one example, controlled release is obtained by appropriate selection of various formulation parameters and ingredients, including, for example, various types of controlled release compositions and coatings. Examples include single or multiple unit tablet or capsule compositions, oil solutions, suspensions, emulsions, microcapsules, microspheres, nanoparticles, patches, and liposomes. In certain embodiments, the composition includes a biodegradable, pH and / or temperature sensitive polymer coating (e.g., enteric coating). Dissolution or diffusion controlled release can be achieved by appropriate coating of tablet, capsule, pellet or granule formulations of the compound, or by incorporating the compound into a suitable matrix. The controlled release coating may include one or more of the coating materials mentioned above.

[0072] There are several techniques known in the art for applying enteric coatings to pharmaceutical compositions. In one non-limiting example, tablets and / or capsules may undergo a drum coating process, in which they are sprayed with coating solution while being rolled in a cylindrical drum. In an additional non-limiting example, tablets and / or capsules may undergo a fluidized bed coating process, in which air is passed over the tablets or capsules at a speed sufficient to separate the tablets and / or capsules into individual units, at which point the tablets and / or capsules are sprayed with coating solution from above (e.g., top spray coating), below (e.g., bottom spray coating), or from the side (e.g., HP spray coating). The amount of coating is determined by the amount of solution sprayed. Coatings may be applied in a single application, or layered through the use of multiple applications.

[0073] Solid Dosage Forms for Oral Administration The pharmaceutical compositions contemplated by the present invention include those formulated for oral administration ("oral dosage forms"). Oral dosage forms can be, for example, in the form of tablets, capsules, liquid solutions or suspensions, powders, or liquid or solid crystals, which contain the active ingredient in admixture with non-toxic pharma- ceutically acceptable excipients. These compositions can be prepared in a variety of ways well known in the pharmaceutical arts, and can be made to release the OXPHOS inhibitor, e.g., the compound of formula 1 or a pharma- ceutically acceptable salt thereof, at a specific segment of the gastrointestinal tract, at a controlled time, with a variety of excipients and formulation techniques. For example, formulations can be tailored to address specific diseases, achieve the plasma levels of the OXPHOS inhibitor, e.g., the compound of formula 1, required to achieve therapeutic efficacy, allow the desired duration of drug effect, and provide a range of compositions with different drug releases.

[0074] OXPHOS inhibition Several techniques for measuring OXPHOS inhibition are known in the art. Exemplary, non-limiting methods for measuring OXPHOS inhibition are described below.

[0075] Measurement of mitochondrial complex I inhibition The electron transport complex in mitochondria is composed of five complexes. Complex I receives electrons from NADH produced by glycolysis and the TCA cycle, and the electrons are transferred to complexes II, III, and IV, which finally transfer the electrons to O2 and produce water molecules. During electron transport, a proton gradient is produced, and the chemical gradient is the driving force for synthesizing ATP at complex V. Mitochondrial inhibition of complex I is indirectly assessed by measuring the oxygen consumption rate (OCR) at complex IV. When the mitochondrial ETC is inhibited, glycolysis is upregulated and lactate production increases. As lactate is transported out of the cell, the extracellular solution becomes acidic (lower pH). OCR and extracellular acidification rate (ECAR) are determined by an XF analyzer (Seahorse Biosciences). Thus, the OXPHOS inhibitory effect of a compound on a sample of cells, such as cancer cells, e.g. the OXPHOS inhibitory effect caused by a compound of formula I, can be measured by measuring the oxygen consumption rate of a sample of cells of the extracellular acidification rate of a sample of cells.

[0076] Cytotoxicity assay under low glucose conditions Inhibition of oxidative phosphorylation (OXPHOS) is not cytotoxic to cells under normal glucose conditions because it is assumed that normal cells have compensatory mechanisms under conditions of energy stress such as low glucose. However, OXPHOS inhibitors show cytotoxic effects on cells under glucose deprivation conditions. In the tumor microenvironment, glucose deprivation conditions are observed, potentially due to poor angiogenesis. Therefore, OXPHOS inhibitors may show anticancer effects on cancer cells under low glucose conditions, which may represent the tumor microenvironment.

[0077] Equivalents and Scope Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments in accordance with the invention described herein. The scope of the present invention is not intended to be limited to the above description.

[0078] The following examples are illustrative only and are not intended to limit the invention in any way. EXAMPLES

[0079] Example 1. Pre- and post-treatment biopsies of patients undergoing cancer therapy Biopsies were collected from subjects undergoing treatment for cancer (Figure 1A). Cells captured in the post-treatment biopsy utilized more OXPHOS than cells captured in the pre-treatment biopsy and utilized less glycolysis than cells captured in the pre-treatment biopsy (Figures 1B-1J). The following was found: 1) ENO1 and SLC45A1 expression are not robust biomarkers of OXPHOS and show positive and negative correlations, respectively, in different cancer types. a) Single copy loss (LOH) of a gene locus is associated with higher OXPHOS levels, particularly in cholangiocarcinoma, where it is common (approximately 33%). 2) A pan-cancer gene-based analysis identified seven genes with higher pathogenic mutation burden associated with increased OxPhos pathway scores (FDR < 5%) - STK11, CDH1, PRKAR1A, KEAP1, TSC2, MAP3K1 and PIK3CA. 3) Single variant analysis tested within each cancer identified seven variants associated with increased OxPhos score with medium effect sizes (FDR<10%) - AKT1, TP53 (two variants) STAG2, PIK3CA, KMT2C and MTOR. 4) A single-variant meta-analysis across cancers identified two variants in the genes RET and TP53 that were detected as contributing in more than once cancer. 5) Most therapies appear to increase OXPHOS (or select for clones with high OXPHOS levels). a) In particular, leuprolide, enzalutamide, fulvestrant, letrozole, palbociclib, cyclophosphamide, carboplatin, doxorubicin, paclitaxel, and temozolomide were the most significant therapies.

[0080] Example 2. Evaluation of the combination of Formula 1 and gemcitabine for treating pancreatic ductal adenocarcinoma Patient-derived xenograft cells were treated with the compound of Formula 1, gemcitabine (Gem), or the compound of Formula 1 and gemcitabine together (Figure 2). Cells treated with both Gem and Formula 1 grew slower than cells treated with Formula 1 or Gem alone.

[0081] Animals with pancreatic ductal adenocarcinoma were treated with either the compound of Formula 1, Gem, or a combination of the compound of Formula 1 and Gem (Figure 3). Animals treated with the combined Gem and compound of Formula 1 survived longer than animals treated with either Gem or the compound of Formula 1 alone.

[0082] Mechanism of action: Expression of biomarkers pAMPK and HIF-1a in untreated cells versus cells treated with the compound of Formula 1 confirms that Gem in combination with Compound 1 inhibits cancer cell growth and extends survival (Figures 4a and 4b).

[0083] Example 3. Clinical Study of Formula 1 in Combination with Standard of Care to Treat BRAF Mutant Metastatic Melanoma in Human Subjects Subjects with BRAF mutant metastatic melanoma are enrolled in a Phase 2 clinical study to evaluate the efficacy of the compound of Formula 1 in combination with standard of care (SOC) for treating BRAF mutant metastatic melanoma (Figure 5). Dose escalation and dose selection will be determined in an initial cohort of 20 subjects receiving SOC, a BRAFi (encorafenib, 450 mg PO QD) and a MEKi (binimetinib (45 mg PO BID)) in combination with the compound of Formula 1 (escalating doses from 100 mg to 200 mg, 400 mg, 800 mg).

[0084] After determining the dose of the compound of Formula 1, the cohort is expanded to 100 subjects. One arm is administered BRAFi, MEKi and the compound of Formula 1, and the second arm is administered BRAFi and MEIKi.

[0085] The primary endpoints were ORR and PFS. Secondary endpoints were safety, tolerability, and OS.

[0086] Example 4. Clinical Study of Formula 1 in Combination with Standard of Care to Treat Pancreatic Ductal Adenocarcinoma in Human Subjects Subjects with pancreatic ductal adenocarcinoma are enrolled in a clinical study to evaluate the efficacy of the compound of Formula 1 in combination with standard of care (SOC) for treating pancreatic ductal adenocarcinoma (Figure 6). Dose escalation and dose selection will be determined in an initial cohort of 12 subjects receiving SOC, gemcitabine and abraxane, in combination with the compound of Formula 1 (400mg>800mg).

[0087] After determining the dose of the Formula 1 compound, the cohort will be expanded to 13 subjects. One arm will receive SOC + Formula 1 compound at the determined dose. The comparator arm will be derived from existing MDACC data of patients with a similar profile to those enrolled in this study.

[0088] The primary endpoints were safety and tolerability. Secondary endpoints were ORR, PFS, and OS.

[0089] Example 5. Evaluation of the response of cell subtypes derived from glioblastoma patients to compounds of Formula 1 Cell subtypes derived from glioblastoma patients were treated with the compound of Formula 1 (FIG. 7A). The MTC GBM cell subtype was more responsive to treatment with the compound of Formula 1 than the GPM cell subtype (FIG. 7B).

[0090] Example 6. Clinical Study of Compound of Formula 1 in Combination with Standard of Care for Treating BRAF Mutant Metastatic Melanoma in Human Subjects A phase 0 onset / withdrawal (window of therapeutic opportunity) study in subjects newly diagnosed with GBM to assess the effect of the compound of Formula 1 on brain penetration and biomarkers will be performed in 12-24 subjects (Figure 8).

[0091] Newly diagnosed subjects with GBM with MTC subtype (N=60) will be enrolled in the study to evaluate the efficacy of the compound of Formula 1 in combination with SOC, radiation / tezolomide, in treating GBM with MTC subtype. One arm will receive SOC alone and the second arm will receive SOC in combination with the compound of Formula 1.

[0092] Subjects (N=20) with BRAF mutant melanoma and brain metastases are enrolled in the study to evaluate the efficacy of the compound of Formula 1 in treating BRAF mutant melanoma and brain metastases. Subjects are administered the compound of Formula 1 at a selected MTD dose.

[0093] The primary endpoints for both the study of subjects with GBM with MTC subtype and the study of patients with BRAF mutant melanoma and brain metastases are ORR and OFS. Secondary endpoints are safety, tolerability, and OS.

[0094] Example 7. Clinical Study of Formula 1 in Combination with Standard of Care to Treat Castration-Resistant Prostate Cancer Subjects (N=30) will be enrolled in the clinical study to evaluate the efficacy of the compound of formula 1 in combination with SOC, a CBI / PD-1 inhibitor, in treating castration-resistant prostate cancer (Figure 9). One arm will receive SOC + compound of formula 1. One arm will receive SOC alone. Primary endpoints are ORR and PFS. Secondary endpoints are safety, tolerability, and OS.

[0095] Example 8. Therapeutic Range of Compounds of Formula 1 The therapeutic window of the compound of formula 1 for various indications was determined (Figure 10).

[0096] Other embodiments Various modifications and variations of the described invention will become apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention that are obvious to those skilled in the art are intended to be within the scope of the present invention.

[0097] All references, patents, patent application publications and patent applications cited herein are hereby incorporated by reference in their entirety.

[0098] Other embodiments are within the claims.

Claims

1. 1. A method of treating cancer in a subject, the method comprising administering an oxidative phosphorylation (OXPHOS) inhibitor to the subject, wherein the cancer is refractory to first-line treatment or recurrent.

2. 10. The method of claim 1, wherein the first line treatment is a standard of care for cancer.

3. 3. The method of claim 1 or 2, wherein the cancer is carcinoma, sarcoma, leukemia or lymphoma.

4. The method of any one of claims 1 to 3, wherein the cancer is a solid tumor.

5. The method of any one of claims 1 to 4, wherein the cancer is a metastatic cancer.

6. 4. The method of claim 3, wherein the leukemia is acute lymphocytic leukemia, chronic lymphocytic leukemia, myeloid leukemia or chronic myelogenous leukemia.

7. 4. The method of claim 3, wherein the lymphoma is Hodgkin's lymphoma or non-Hodgkin's lymphoma.

8. 4. The method of claim 3, wherein the cancer is breast cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, bladder cancer, colon cancer, rectal cancer, uterine cancer, vaginal or vulvar cancer, testicular cancer, kidney cancer, liver cancer, bile duct cancer, cholangiocarcinoma, gallbladder cancer, melanoma, pancreatic cancer, pancreatic ductal adenocarcinoma, BRAF mutant metastatic melanoma, prostate cancer, castration resistant prostate cancer, thyroid cancer, glioblastoma, endometrial cancer, ovarian cancer, cervical cancer, brain cancer, glioblastoma, liver cancer, colorectal cancer, skin cancer, basal cell carcinoma, squamous cell carcinoma, melanoma, gynecological cancer, head and neck cancer, mesothelioma, myeloma, esophageal cancer or gastric cancer.

9. The OXPHOS inhibitor is a compound of formula 1 below or a pharma- ceutically acceptable salt thereof: The method according to any one of claims 1 to 8, wherein

10. 10. The method of claim 9, wherein the compound of formula 1 is formulated as a tablet or capsule.

11. 10. The method of claim 9, wherein the tablet or capsule is formulated for delayed release.

12. 12. The method of claim 10 or 11, wherein the tablet or capsule comprises an enteric coating.

13. 13. The method of any one of claims 9 to 12, wherein the compound of formula 1 is administered at a dose of about 100 mg, 200 mg, 400 mg, 800 mg or 1200 mg per day.

14. 14. The method of claim 13, wherein the compound of formula 1 is administered at a dosage of about 800 mg per day.

15. 10. The method of any one of claims 1 to 9, wherein the OXPHOS inhibitor is administered in combination with a second therapeutic compound.

16. A method for treating pancreatic ductal adenocarcinoma (PDAC), comprising administering to a subject in need thereof a therapeutically effective dose of a pharmaceutical composition, the composition comprising as an active ingredient a compound of formula 1 or a pharma- ceutical acceptable salt thereof: A method comprising:

17. 17. The method of claim 16, wherein the compound of formula 1 is administered in combination with gemcitabine and abraxane.

18. A method for treating BRAF mutant melanoma, the method comprising administering to a subject in need thereof a therapeutically effective dose of a pharmaceutical composition, the composition comprising as an active ingredient a compound of formula 1 or a pharma- ceutical acceptable salt thereof: A method comprising:

19. 19. The method of claim 18, wherein the compound of formula 1 is administered in combination with encorafenib and binimetinib.

20. 20. The method of claim 18 or 19, wherein the BRAF mutant melanoma is metastatic.

21. 21. The method of claim 20, wherein the metastasis is to the central nervous system.

22. 22. The method of claim 20 or 21, wherein the metastasis is to the brain.

23. 23. The method of claim 22, comprising the use of radiation and / or the administration of temozolomide.

24. A method for treating castration-resistant prostate cancer (CRPC), comprising administering to a subject in need thereof a therapeutically effective dose of a pharmaceutical composition, the composition comprising, as an active ingredient, a compound of formula 1 or a pharma- ceutical acceptable salt thereof: A method comprising:

25. 25. The method of claim 24, wherein the subject has failed to respond to at least one course of immunotherapy.

26. 26. The method of claim 24 or 25, wherein the compound of formula 1 is administered in combination with a checkpoint inhibitor.

27. 27. The method of claim 26, wherein the checkpoint inhibitor is a PD-1 inhibitor.

28. 28. The method of any one of claims 16 to 27, wherein the compound of formula 1 is formulated as a tablet or capsule.

29. 30. The method of claim 28, wherein the tablet or capsule is formulated for delayed release.

30. 30. The method of claim 28 or 29, wherein the tablet or capsule comprises an enteric coating.

31. 31. The method of any one of claims 16 to 30, wherein the compound of formula 1 is administered at a dose of about 100 mg, 200 mg, 400 mg, 800 mg, or 1200 mg per day.

32. 32. The method of claim 31, wherein the compound of formula 1 is administered at a dosage of about 800 mg per day.

33. 33. The method of any one of claims 9 to 32, wherein the pharma- ceutically acceptable salt is a salt of an acid selected from the group consisting of formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, trifluoroacetic acid, malic acid, maleic acid, malonic acid, fumaric acid, succinic acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, dichloroacetic acid, aminooxyacetic acid, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, and boric acid.

34. 34. The method of claim 33, wherein the pharma- ceutically acceptable salt is a salt of acetic acid.

35. 35. The method of any one of claims 1 to 34, wherein the subject is a human.

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