Methods for Treating Mitochondrial-Related Disorders

Abstract

Disclosed herein is a method of treatment comprising administering a therapeutically effective amount of 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharma- ceutically acceptable salt thereof, to a subject in need thereof. 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole is useful for treating mitochondrial-related disorders or conditions, including obesity, diabetes, hypertension, cardiovascular disease, and liver disease.

Description

[Technical field]

[0001] The present disclosure relates to methods for treating cardiovascular disease and mitochondrial-related disorders or conditions without causing clinically significant risks of adverse events or overdose. [Background technology]

[0002] Cardiovascular disease is a type of disease involving the heart and blood vessels, including coronary artery disease, heart attack, stroke, heart failure, hypertensive heart disease, and rheumatic heart disease. More than 6.5 million people in the United States suffer from heart failure (Cardiology Today, April 6, 2017). Heart failure with preserved ejection fraction (HFpEF), also known as diastolic heart failure, causes nearly half of the 6.5 million heart failure cases in the United States. HFpEF results from abnormalities in active ventricular relaxation and passive ventricular compliance, resulting in reduced stroke volume and cardiac output (Am Fam Physician. 2017 Nov 1; 96(9):582-588). In heart failure with reduced ejection fraction (HFrEF), also known as systolic heart failure, the heart muscle cannot contract adequately, resulting in less oxygen-rich blood being pumped out of the body. Mortality rates were similar in HFpEF and HFrEF patients (Cardiology Today, April 6, 2017). Heart failure with mildly reduced ejection fraction (HFmrEF) is a new category of heart failure that is intermediate between HFpEF and HFrEF. The prevalence of HFmrEF is 10-20% of heart failure patients (Maedica (Bucur), 2016, 11 (4): 320-324). Therefore, there is a great need for effective treatment of heart failure, including HFpEF, HFrEF, and HFmrEF.

[0003] Mitochondria control the metabolism of individual cells by burning sugars and fats. Mitochondrial uncoupling is a robust and natural process that the body uses to generate heat. Heat is generated in mitochondria through the uncoupling of respiration (complexes I-IV) and ATP phosphorylation (complex V). In fact, 20-40% of calories consumed are used to generate body heat. Mitochondrial-related disorders or conditions occur when mitochondria are unable to produce enough energy to function properly and can affect almost every part of the body, including cells of the brain, adipose tissue, nerves, muscles, heart, lungs, liver, kidneys, pancreas, eyes, and ears.

[0004] Administering chemical uncouplers for mitochondria as a means to reduce fat deposits has been a scientific goal for many years. There are several small molecules that uncouple mitochondrial oxidative phosphorylation, the best known being 2,4-dinitrophenol (DNP). DNP is known to uncouple with great potency, but unfortunately, it is associated with an unacceptably high rate of serious side effects (J.Med.Toxicol.2011 Sep;7(3):205-212). These side effects include hyperthermia, tachycardia, sweating and tachypnea, and ultimately death. DNP, a small, highly penetrating, lipophilic acid, is rapidly absorbed in the stomach. High concentrations are rapidly distributed and immediately uncouple, producing high levels of heat in a short period of time. Thus, DNP has a small therapeutic index and is very dangerous if overdosed. DNP was deemed "highly dangerous and unfit for human consumption" by the US Federal Food, Drug and Cosmetic Act of 1938. Accordingly, there is a need for uncouplers that can safely treat mitochondrial-associated disorders or conditions.

[0005] 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole is a novel small molecule uncoupler (compound 1). It acts as a controlled metabolic enhancer (CMA). CMA is designed to effectively address the accumulation of fat and sugar in the body, which is the root cause of metabolic disease. CMA works to improve cellular metabolism, increase energy expenditure and calorie expenditure, and reduce fat accumulation. Through a novel controlled and targeted manner, compound 1 can increase mitochondrial proton leak. Mitochondrial proton leak is an ongoing process in the body that dissipates energy, accounting for 20%-40% of daily calories. Compound 1 utilizes the mitochondrial uncoupling mechanism to increase substrate utilization. Compound 1 has been studied in preclinical models. Its potent therapeutic activity in relevant rodent disease models, together with its pharmacokinetic and safety profile, supports that compound 1 may be beneficially and safely used to treat a wide range of mitochondrial-related diseases. Compound 1 may also be effective in treating cardiovascular diseases. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Am Fam Physician.2017 Nov 1;96(9):582-588 [Non-Patent Document 2] Maedica(Bucur),2016,11(4):320-324 [Non-Patent Document 3] J.Med.Toxicol.2011 Sep;7(3):205-212 Summary of the Invention [Means for solving the problem]

[0007] In some embodiments, the disclosure provides a method of treating cardiovascular disease in a subject, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutically acceptable salt thereof.

[0008] In some embodiments, the disclosure provides a method of treating heart failure, comprising administering to a subject a therapeutically effective amount of Compound 1, or a pharma- ceutically acceptable salt thereof.

[0009] In some embodiments, the disclosure provides a method for reducing cardiovascular risk or mortality in a subject suffering from a cardiovascular disease condition, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0010] In some embodiments, the cardiovascular disease is selected from the group consisting of heart failure, heart attack, coronary artery disease, and coronary heart disease (CHD).

[0011] Heart failure as used herein includes heart failure with preserved ejection fraction (HFpEF), or heart failure with reduced ejection fraction (HFrEF), or heart failure with mildly reduced ejection fraction (HFmrEF).

[0012] In some embodiments, the cardiovascular disease is HFpEF.

[0013] In some embodiments, the cardiovascular disease is HFrEF.

[0014] In some embodiments, the cardiovascular disease is HFmrEF.

[0015] In some embodiments, the disclosure provides a method for treating heart failure with preserved ejection fraction (HFpEF) in a subject, the method comprising administering to the subject a therapeutically effective amount of 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharma- ceutically acceptable salt thereof.

[0016] In some embodiments, the disclosure provides a method for treating heart failure with reduced ejection fraction (HFrEF) in a subject, the method comprising administering to the subject a therapeutically effective amount of 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharma- ceutical acceptable salt thereof.

[0017] In some embodiments, the disclosure provides a method for treating heart failure with mildly reduced ejection fraction (HFmrEF) in a subject, the method comprising administering to the subject a therapeutically effective amount of 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharma- ceutical acceptable salt thereof.

[0018] In some embodiments, the subject suffers from at least one of shortness of breath, shortness of breath on exertion, impaired cardiac energy, dizziness, fatigue, dyspnea, palpitations (atrial fibrillation), chest discomfort, edema, fainting, and limitations in activities of daily living.

[0019] In some embodiments, limitations in activities of daily living are difficulties with personal care, mobility, and eating.

[0020] In some embodiments, the subject suffers from a symptom selected from: decreased exercise tolerance, fatigue, lethargy, increased recovery time after exercise, and swollen ankles.

[0021] In some embodiments, the subject suffers from at least one of coronary artery disease, hypertension, and a heart murmur.

[0022] In some embodiments, the disclosure provides a method of lowering blood pressure in a subject, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutically acceptable salt thereof.

[0023] In some embodiments, the subject suffers from at least one of cardiovascular disease, hypertension, resistant hypertension, and severe hypertension.

[0024] In some embodiments, the cardiovascular disease is heart failure (which may be HFpEF, HFrEF, or HFmrEF), heart attack, coronary artery disease, or coronary heart disease (CHD).

[0025] In some embodiments, the subject has hypertension associated with HFpEF.

[0026] In some embodiments, the subject has hypertension associated with HFrEF.

[0027] In some embodiments, the subject has hypertension associated with HFmrEF.

[0028] In some embodiments, the subject experiences a reduction in blood pressure of at least 5 mmHg following administration.

[0029] In some embodiments, the methods reduce the risk of developing cardiovascular disease and / or reduce the risk of HFpEF, HFrEF, or HFmrEF.

[0030] In some embodiments, the method slows the progression of HFpEF, HFrEF, or HFmrEF.

[0031] In some embodiments, the method comprises: i) The half-life of DNP (t 1 / 2 ) to be extended; ii) Time to maximum plasma concentration of DNP (T max ) to delay; iii) The peak plasma concentration of DNP (C max ) ; and iv) Increasing the area under the curve (AUC) Includes at least one of the following:

[0032] In some embodiments, the subject experiences no significant systemic toxicity, side effects, significant elevation in body temperature, or significant elevation in heart rate following administration.

[0033] In some embodiments, the present disclosure provides: i) DNP peak plasma concentration (C max ) steady state; ii) an average half-life (t 1 / 2 ); iii) The peak plasma concentration of DNP (T max ) median time to reach; iv) Area under the curve (AUC) extrapolated to infinity for DNP from about 3 h*μg / mL to about 420 h*μg / mL inf ) median; and v) AUC / C of about 18 max ratio The present invention provides a method for treating cardiovascular disease by administering about 30 mg to about 1,400 mg of compound 1 or a pharma- ceutical acceptable salt thereof to a subject, thereby achieving the above-mentioned objective.

[0034] In some embodiments, the disclosure provides a method of treating a mitochondrial-related disorder or condition in a subject without causing a clinically significant risk of an adverse event, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0035] In some embodiments, the disclosure provides a method for reducing toxicity or side effects in treating a mitochondrial-related disorder or condition in a subject, the method comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0036] In some embodiments, the disclosure provides a method for preventing overdosing in treating a mitochondrial-related disorder or condition in a subject, the method comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0037] In some embodiments, the disclosure provides a method for increasing metabolic rate or resting energy expenditure in a subject without causing a clinically significant risk of adverse events, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0038] In some embodiments, the disclosure provides a method for treating a metabolic disorder in a subject, the method comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutically acceptable salt thereof.

[0039] In some embodiments, the disclosure provides a method for treating hypertriglyceridemia in a subject associated with cardiovascular disease, atherosclerosis, obesity, hypertension, diabetes, insulin resistance, and / or liver disease, comprising administering a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof, to the subject.

[0040] In some embodiments, the disclosure provides a method for treating severe hypertriglyceridemia in a subject, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0041] In some embodiments, the disclosure provides a method for reducing liver fat by at least 50%, or reducing lipids by at least 10%, in a subject, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0042] In some embodiments, the disclosure provides a method of treating obesity, excess body fat, type 2 diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, atherosclerosis, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, Rett syndrome, metabolic syndrome associated with aging, metabolic diseases associated with increased reactive oxygen species (ROS), Friedreich's ataxia, NAFLD, NASH, non-sclerotic NASH, non-sclerotic NASH with liver fibrosis, hepatic steatosis, hepatic fibrosis, cirrhosis, or hepatocellular carcinoma, comprising administering a therapeutically effective amount of Compound 1, or a pharmacologic acceptable salt thereof, to a subject; i) Peak plasma concentrations of 2,4-dinitrophenol (C max ) steady state; ii) an average half-life (t 1 / 2 ); iii) The peak plasma concentration of 2,4-dinitrophenol (T max ) median time to reach; iv) Area under the curve (AUC) extrapolated to infinity for 2,4-dinitrophenol from about 3 h*μg / mL to about 420 h*μg / mL inf ) median; and v) AUC / C of about 18 max ratio To provide a method for achieving at least one of the above. [Brief description of the drawings]

[0043] [Figure 1] 1 shows plasma concentrations of 2,4-dinitrophenol following administration of Compound 1 and 2,4-dinitrophenol to dogs.

[0044] [Diagram 2] A and B show the AUC of 2,4-dinitrophenol after administration of Compound 1.

[0045] [Diagram 3] A and B show plasma concentrations of Compound 1 following oral administration of 500 mg of Compound 1 by food state (linear and semi-log scale).

[0046] [Figure 4] A and B show plasma concentrations of 2,4-dinitrophenol following oral administration of 500 mg of Compound 1 by food state (linear and semi-log scale).

[0047] [Diagram 5] 2,4-Dinitrophenol curves for the entire cohort are shown.

[0048] [Figure 6] 1 shows body temperatures of the MAD cohort during QD dosing of MAD.

[0049] [Figure 7] Shown is the resting energy expenditure (REE) of the MAD cohort during QD administration of MAD.

[0050] [Figure 8] 4 shows the weight loss after administration of Compound 1.

[0051] [Figure 9] 1 shows the reduction in mean glucose levels following administration of Compound 1.

[0052] [Figure 10] 4 shows the change in systolic blood pressure after administration of Compound 1.

[0053] [Figure 11] 4 shows the change in diastolic blood pressure after administration of Compound 1.

[0054] [Figure 12] The heart rates observed after administration of Compound 1 are shown.

[0055] [Figure 13] 4 shows the changes in body temperature observed after administration of Compound 1.

[0056] [Figure 14] PK results observed following administration of Compound 1 are shown.

[0057] [Figure 15] Figure 1 shows the placebo-corrected percent change from baseline values ​​in MRI-proton density fat fraction (PDFF) following administration of Compound 1.

[0058] [Figure 16] 1 shows an analysis of covariance mean change from baseline for MRI-proton density fat fraction (PDFF) at day 61 in the FAS population.

[0059] [Figure 17] Mean change from baseline FAS population in repeated measures analysis of InBody weight is shown.

[0060] [Figure 18] The observed changes in body weight after administration of Compound 1 are shown.

[0061] [Figure 19] 1 shows placebo-corrected percent change from baseline in abdominal MRI liver volume and fat fraction by treatment group FAS population.

[0062] [Figure 20] 1 shows placebo-corrected percent change from baseline in abdominal MRI liver volume and fat fraction by treatment group FAS population.

[0063] [Figure 21] MRI confirmed fat loss (total adipose tissue).

[0064] [Figure 22] The observed changes in liver volume are shown.

[0065] [Figure 23] The mean change from baseline in systolic blood pressure observed at day 61 is shown.

[0066] [Figure 24] The mean change from baseline in diastolic blood pressure observed at day 61 is shown.

[0067] [Diagram 25] The mean change from baseline in high sensitivity C-reactive protein (hsCRP) observed at day 61 is shown. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0068] definition While various embodiments and aspects of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments and aspects are provided by way of example only. Numerous variations, changes, and substitutions will occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be utilized in practicing the invention.

[0069] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. All documents or portions of documents cited in this application, including but not limited to patents, patent applications, papers, books, manuals, and articles, are expressly incorporated herein by reference in their entirety for all purposes.

[0070] 5-[(2,4-Dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole is a novel small molecule uncoupler and has the following structure: [ka]

[0071] 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole can be prepared by the procedure described in WO2018 / 129258.

[0072] In this disclosure, compound 1 and CM1 are interchangeable, and both refer to 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole.

[0073] Unless otherwise specified, technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. See, for example, Singleton et al., DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY, 2nd ed., J.Wiley & Sons (New York, NY 1994); Sambrook et al., MOLECULAR CLONING, A LABORATORY MANUAL, Cold Springs Harbor Press (Cold Springs Harbor, NY 1989). Any methods, devices and materials similar or equivalent to those described herein can be used to practice the present invention. The following definitions are provided to facilitate understanding of certain terms frequently used herein and are not intended to limit the scope of the present disclosure.

[0074] The terms "a" or "an" as used herein mean one or more.

[0075] The terms "comprise," "include," "having," and their derivatives are used interchangeably herein as inclusive, open-ended terms. For example, the use of "comprising," "including," or "having" means that whatever element is comprised of, held, or included is not the only element contained in the subject of the clause containing the verb.

[0076] As used herein, the term "about" refers to a range of values ​​that includes a particular value and that one of ordinary skill in the art would consider to be reasonably similar to the particular value. In some embodiments, the term "about" refers to within a range of standard deviation using measurements generally accepted in the art. In some embodiments, "about" refers to a range that covers ±10%, ±5%, or ±2% of the particular value. In some embodiments, "about" refers to the particular value.

[0077] As used herein, "treatment", "treat", "alleviate", "ameliorate" or "alleviate" are used interchangeably. These terms refer to a method for obtaining a beneficial or desired result, including but not limited to therapeutic benefit. Therapeutic benefit refers to eradication or amelioration of the underlying disease being treated. Therapeutic benefit is also achieved by eradication or amelioration of one or more physiological symptoms associated with the underlying disease, such that an improvement is observed in the subject, even though the subject may still be suffering from the underlying disease. Treatment includes delaying the onset of clinical symptoms of the disease by administration of the composition; suppressing the disease, i.e., causing a reduction in clinical symptoms of the disease; inhibiting the disease, i.e., preventing the onset of clinical symptoms by administration of the composition after the first appearance of symptoms; and / or alleviating the disease, i.e., causing a regression of clinical symptoms by administration of the composition after the first appearance of clinical symptoms.

[0078] "Patient", "subject", or "subject in need thereof" refers to an organism suffering from or susceptible to a disease or condition that can be treated using the methods provided herein. The term does not necessarily indicate that the subject has been diagnosed with a particular disease, but typically refers to an individual under medical care. Non-limiting examples include humans, other mammals, cows, rats, mice, dogs, cats, monkeys, goats, sheep, cows, deer, and other non-mammals. In some embodiments, the patient, subject, or subject in need thereof is a human.

[0079] As used herein, "administration" of the disclosed compounds includes delivering a compound as described herein, or a prodrug or other pharma- ceutically acceptable derivative thereof, to a subject using any suitable formulation or route of administration, e.g., as described herein.

[0080] "Pharmaceutically acceptable" refers to compounds, salts, compositions, dosage forms and other materials that are useful in preparing pharmaceutical compositions suitable for animal or human pharmaceutical use.

[0081] As used herein, "pharmaceutically acceptable salts" refers to salts of administered compounds prepared from pharmaceutically acceptable non-toxic acids and bases, including inorganic acids, inorganic bases, organic acids, inorganic bases, solvates, hydrates and clathrates thereof. Suitable pharmaceutically acceptable acid addition salts can be prepared from inorganic or organic acids. Examples of inorganic acids include sulfate, hydrogen sulfate, hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, sulfuric acid and phosphoric acid (including hydrogen phosphate and dihydrogen phosphate). Suitable organic acids may be selected from aliphatic, alicyclic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic organic acids, examples of which include formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, gluconic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, glucuronic acid, maleic acid, fumaric acid, pyruvic acid, aspartic acid, glutamic acid, benzoic acid, anthranilic acid, 4-hydroxybenzoic acid, phenylacetic acid, mandelic acid, embonic acid (pamoic acid), methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, pantothenic acid, trifluoromethanesulfonic acid, 2-hydroxyethanesulfonic acid, p-toluenesulfonic acid, sulfanilic acid, cyclohexylaminosulfonic acid, stearic acid, alginic acid, β-hydroxybutyric acid, salicylic acid, galactaric acid and galacturonic acid. Suitable pharma- ceutically acceptable base addition salts of the compounds of the present invention include, for example, ammonium salts and metal salts including alkali metal salts, alkaline earth metal salts and transition metal salts, such as, for example, calcium salts, magnesium salts, potassium salts, sodium salts and zinc salts. Pharmaceutically acceptable base addition salts also include organic salts prepared from basic amines such as, for example, N,N'-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine), and procaine. All of these salts can be prepared from the corresponding compound, for example, by reacting the appropriate acid or base with the compound.

[0082] An "effective amount" is an amount sufficient to achieve a stated purpose (e.g., achieve the effect for which it is administered, treat a disease, reduce enzyme activity, alleviate one or more symptoms of a disease or condition, reduce viral replication in cells). Examples of an "effective amount" include an amount sufficient to contribute to the treatment of a disease or the alleviation of a symptom(s), which may also be referred to as a "therapeutically effective amount". "Alleviation" of a symptom(s) (and grammatical equivalents of this phrase) means a decrease in the severity or frequency of the symptom(s), or the elimination of the symptom(s). Efficacy can also be expressed as a "fold" increase or a "fold" decrease. For example, a therapeutically effective amount may have at least a 1.2-fold, 1.5-fold, 2-fold, 5-fold, or more effect over a control.

[0083] As used herein, the term "significant increase in body temperature" in a subject refers to an increase in body temperature that is associated with adverse effects on the subject, including but not limited to illness, physical discomfort or pain, coma, and death. In one non-limiting embodiment, the significant increase in body temperature is an increase of about 0.5°C, about 1°C, about 1.5°C, about 2°C, about 2.5°C, about 3°C, about 3.5°C, about 4°C, about 4.5°C, about 5°C, about 5.5°C, about 6°C, or more. In another non-limiting embodiment, the significant increase in body temperature lasts for about 5 minutes, about 15 minutes, about 30 minutes, about 45 minutes, about 1 hour, about 1.5 hours, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, about 24 hours, or more.

[0084] As used herein, the term "significant systemic toxicity" in a subject refers to systemic toxicity that is associated with adverse effects to the subject, including but not limited to illness, physical discomfort or pain, coma, and death. In one non-limiting embodiment, significant systemic toxicity is indicated by an increase in liver enzyme, blood urea nitrogen, or creatinine levels compared to the corresponding levels in the subject when the composition is not administered.

[0085] Treatment method In some embodiments, the disclosure provides a method of treating cardiovascular disease in a subject, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof. Initial studies have shown that Compound 1: Controlled and improved PK-low C of 2,4-dinitrophenol max , high AUC, low variability, Possibility of once-daily oral administration; Liver-specific uncoupling and reduced systemic effects; Broad therapeutic index Thus, compound 1 is useful for safely treating a wide range of diseases.

[0086] In some embodiments, the disclosure provides a method for treating heart failure in a subject suffering from symptoms due to cardiovascular disease, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0087] In some embodiments, the disclosure provides a method for reducing cardiovascular risk or mortality in a subject suffering from a cardiovascular disease condition, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0088] In some embodiments, the symptoms of cardiovascular disease are shortness of breath, dizziness, chest pain, fainting, fatigue, or limitation in activities of daily living.

[0089] In some embodiments, a limitation in an activity of daily living is difficulty with personal care, mobility, or eating.

[0090] In some embodiments, the disclosure provides a method of treating cardiovascular disease in a subject, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutically acceptable salt thereof.

[0091] In some embodiments, the cardiovascular disease is associated with obesity. In some embodiments, the cardiovascular disease includes the following diseases, disorders, or conditions:

[0092] Hemodynamic disturbances of the cardiovascular system characterized by increased heart rate in sedentary individuals, increased risk of atrial fibrillation, increased blood volume, increased cardiac output, increased systemic vascular resistance in individuals with hypertension and insulin resistance, increased arterial pressure, increased left ventricular wall stress, increased pulmonary artery pressure, and altered intraventricular pressure in individuals with sleep apnea syndrome.

[0093] Atherosclerosis and myocardial infarction, which can be increased indirectly by promoting major atherosclerotic risk factors (diabetes, hypertension, dyslipidemia, etc.) or directly by adipose endocrine and immune dysregulation of epicardial adipose tissue.

[0094] Epicardial fat accumulation, pathogenic paracrine and vascular endocrine signaling, increased inflammatory macrophages, increased T lymphocytes and mast cells, increased proadiogenic adipokines, and decreased vasoprotective adipokines.

[0095] Heart failure (HF), particularly HF with preserved ejection fraction (HFpEF).

[0096] Atherosclerotic cardiovascular disease (ASCVD), arrhythmias, fatty infiltration of the heart, and increased coronary artery calcium.

[0097] Sleep apnea can lead to hypoxia, increased epinephrine (adrenaline) can lead to high blood pressure, and fluctuations in thoracic pressure increase left and right ventricular pressure.

[0098] Thrombosis and thromboembolic events: Increased adipose tissue compresses pelvic and lower extremity veins, impairing venous return and promoting deep vein thrombosis.

[0099] Cardiac cellular and structural abnormalities characterized by myocardial steatosis, apoptosis and fibrosis, as well as left ventricular remodeling and hypertrophy, left atrial hypertrophy, right ventricular hypertrophy, and increased pericardial and perivascular adipose tissue.

[0100] Sleep apnea leads to hypoxia, atherosclerosis, thrombosis, left ventricular dysfunction (both diastolic and systolic), and reduced cardiac function characterized by right ventricular failure.

[0101] Immune disorders characterized by an increase in pro-inflammatory adipocytokines, such as tumor necrosis factor (TNF), interleukins such as interleukin-6 (IL-6), monocyte chemotactic protein-1 (MCP-1) or C-reactive protein (CRP), or a decrease in anti-inflammatory adipocytokines (e.g., adiponectin) and IL-10.

[0102] An immune disorder characterized by severe asthma-like neutrophil activation and increased granulation tissue formation, as well as glucocorticoid-resistant severe asthma.

[0103] An endocrine disorder characterized by activation of the renin-angiotensin-aldosterone system leading to elevated blood pressure and altered expression of peroxisome proliferator-activated receptors.

[0104] An endocrine disorder characterized by hyperinsulinemia, systemic insulin resistance and steatosis, and myocardial insulin insensitivity.

[0105] An endocrine disorder characterized by leptin insensitivity with elevated leptin levels that may contribute to cardiac hypertrophy and heart failure.

[0106] Lipotoxicity, characterized by limited energy stores in peripheral subcutaneous adipose tissue.

[0107] Excessive supply of free fatty acids to the liver, muscle, pancreas, kidney and / or visceral, pericardial and perivascular adipose tissue.

[0108] In some embodiments, the cardiovascular disease is heart failure, heart attack, coronary artery disease, and coronary heart disease (CHD).

[0109] In some embodiments, the heart failure comprises HFpEF, HFrEF, or HFmrEF.

[0110] In some embodiments, the subject experiences a reduced risk of serious cardiovascular events following administration.

[0111] In some embodiments, a serious cardiovascular event is death or hospitalization due to worsening of the disease.

[0112] In some embodiments, the disclosure provides a method for treating heart failure with preserved ejection fraction (HFpEF) in a subject, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0113] In some embodiments, the disclosure provides a method for treating heart failure with reduced ejection fraction (HFrEF) in a subject, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0114] In some embodiments, the disclosure provides a method for treating heart failure with mildly reduced ejection fraction (HFmrEF) in a subject, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0115] In some embodiments, the subject suffers from obesity, excess body fat, diabetes, high blood pressure (hypertension), dyslipidemia, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, or metabolic syndrome.

[0116] In some embodiments, the subject suffers from at least one symptom selected from shortness of breath, shortness of breath on exertion, impaired cardiac energy, dizziness, fatigue, dyspnea, palpitations (atrial fibrillation), chest discomfort, edema, fainting, and limitation in activities of daily living.

[0117] In some embodiments, limitations in activities of daily living are difficulties with personal care, mobility, and eating.

[0118] In some embodiments, the subject suffers from at least one of decreased exercise tolerance, fatigue, lethargy, increased recovery time after exercise, and ankle swelling.

[0119] In some embodiments, the subject suffers from at least one of coronary artery disease, hypertension, and a heart murmur.

[0120] In some embodiments, the subject experiences improvement in cardiac bioenergetic deficiency following administration, where improvement comprises greater than 5% weight loss, lower blood pressure, improved quality of life, increased exercise tolerance, and / or reduced risk of serious cardiovascular events, where serious cardiovascular events are selected from the group consisting of death, hospitalization for worsening disease, and myocardial infarction.

[0121] In some embodiments, the method further comprises assessing peak oxygen consumption (VO2) and / or slope of VE / CO2 or VE / VCO2 in the subject during exercise before and after administration of a therapeutically effective amount of Compound 1, wherein an increase in VO2 in the subject after administration indicates a decrease in the degree of HFpEF, HFrEF, HFmrEF, or one or more symptomatic components or symptoms of these cardiovascular diseases in the subject.

[0122] In some embodiments, the method increases VO2 in a subject following administration.

[0123] In some embodiments, the method increases exercise capacity in a subject.

[0124] In some embodiments, the method increases the exercise capacity of a subject as measured by assessing the 6-minute walking distance (6MWD) before and after administration of a therapeutically effective amount of Compound 1, wherein an increase in the subject's 6MWD after administration indicates a reduction in the degree of HFpEF, or at least one of its symptomatic components or symptoms, in the subject.

[0125] In some embodiments, the method increases the 6MWD following administration.

[0126] In some embodiments, HFpEF in a subject is diagnosed in light of echocardiography (E / e') or biomarkers (NT-proBNP).

[0127] In some embodiments, the method further comprises assessing the subject's NYHA classification score before and after administration.

[0128] The NYHA functional classification categorizes the severity of heart failure symptoms into one of four functional categories. It is widely used in clinical practice and research because it provides a standard description of severity to assess response to treatment and guide management. The NYHA functional classification, based on the severity of symptoms and physical activity, is as follows: Class I: No limitation of physical activity. Normal physical activity does not cause undue shortness of breath, fatigue or palpitations. Class II: Some limitation in physical activity. Comfortable at rest, but ordinary physical activity causes excessive shortness of breath, fatigue or palpitations. Class III: Severe limitation of physical activity. Comfortable at rest, but less than normal physical activity causes excessive shortness of breath, fatigue or palpitations. Class IV: You are unable to perform any physical activity without discomfort. Symptoms may occur at rest. Any physical activity increases the discomfort.

[0129] In some embodiments, the method further comprises assessing the subject's NYHA classification score before and after administration of a therapeutically effective amount of Compound 1, wherein a decrease in the NYHA score after administration indicates a decrease in the extent of disease in the subject.

[0130] In some embodiments, the method reduces the subject's NYHA classification score from class III to class II, or from class II to class I following administration.

[0131] In some embodiments, the method improves the quality of life of the subject.

[0132] In some embodiments, the methods improve the subject's quality of life as assessed by a standardized questionnaire, such as the Kansas City Cardiomyopathy Questionnaire (KCCQ), KCCQ-12, KCCQ-Physical Limitation Score (KCCQ-PLS), KCCQ-Totally Symptom Score (KCCQ-TSS), KCCQ-Clinical Summary Score (KCCQ-CSS), KCCQ-Overall Summary Score (KCCQ-OSS), or other derivatives.

[0133] In some embodiments, the disclosure provides a method of lowering blood pressure in a subject, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutically acceptable salt thereof.

[0134] In some embodiments, the subject suffers from at least one of cardiovascular disease, hypertension, resistant hypertension, and severe hypertension.

[0135] In some embodiments, the cardiovascular disease is selected from the group consisting of heart failure, HFpEF, HFrEF, heart attack, coronary artery disease, and coronary heart disease (CHD).

[0136] In some embodiments, the subject has hypertension associated with HFpEF.

[0137] In some embodiments, the subject has hypertension associated with HFrEF.

[0138] In some embodiments, the subject has hypertension associated with HFmrEF.

[0139] In some embodiments, the subject is suffering from at least one symptom selected from headache, shortness of breath, chest pain, nosebleed, dizziness, fatigue, blurred vision, irregular heartbeat, blood in the urine, sweating, sleep disorders, and blood spots on the eyes.

[0140] In some embodiments, the condition is associated with HFpEF, HFrEF, or HFmrEF.

[0141] In some embodiments, reducing blood pressure includes reducing diastolic blood pressure and / or reducing systolic blood pressure.

[0142] In some embodiments, the subject experiences a reduction in blood pressure of at least 5 mmHg following administration.

[0143] In some embodiments, the method reduces the risk of developing cardiovascular disease, reduces the risk of HFpEF, or slows the progression of HFpEF.

[0144] In some embodiments, the methods reduce the risk of developing cardiovascular disease, reduce the risk of HFrEF, or slow the progression of HFrEF.

[0145] In some embodiments, the methods reduce the risk of developing cardiovascular disease, reduce the risk of HFmrEF, or slow the progression of HFmrEF.

[0146] In some embodiments, the subject is in a fasted state prior to administration.

[0147] In some embodiments, the subject is in a fed state prior to administration.

[0148] In some embodiments, the subject experiences a decrease in at least one of weight, blood pressure, and blood glucose following administration.

[0149] In some embodiments, the subject experiences at least one of the following: i) at least 5% or at least 10% weight loss; ii) a reduction in blood pressure of at least 5 mmHg; iii) HbA of at least 0.5% or at least 1.5% 1c Decrease in; iv) at least 10% lipid reduction; and v) A reduction in liver fat of at least 50%.

[0150] In some embodiments, the method comprises: The half-life of 2,4-dinitrophenol (t 1 / 2 ) to be extended; Time to maximum plasma concentration of 2,4-dinitrophenol (T max ) to delay; The maximum plasma concentration of 2,4-dinitrophenol (C max ) ; and Increasing the area under the curve (AUC) Includes at least one of the following:

[0151] In some embodiments, the average half-life of 2,4-dinitrophenol is increased to about 20-50 hours, 25-40 hours, or 30-40 hours.

[0152] In some embodiments, the T max The median time to saturation is extended to at least 6 hours or at least 8 hours.

[0153] In some embodiments, the T max The median time is extended to about 6 to 8 hours or about 6 to 10 hours.

[0154] In some embodiments, the C of 2,4-dinitrophenol max Reducing the C of 2,4-dinitrophenol of about 80 ng / mL to about 8300 ng / mL in subjects after administration max This includes providing a steady state of

[0155] In some embodiments, the method provides a method for determining whether a subject has an AUC / C max Ratios are provided.

[0156] In some embodiments, the subject experiences no significant systemic toxicity, side effects, significant elevation in body temperature, or significant elevation in heart rate following administration.

[0157] In some embodiments, side effects include at least one of nausea, vomiting, sweating, dizziness, headache, cataracts, glaucoma, fever, hyperthermia, tachycardia, hyperhidrosis, tachypnea, and death.

[0158] In some embodiments, the disclosure provides a method of treating cardiovascular disease, comprising administering to a subject about 30 mg to about 1400 mg of Compound 1, or a pharma- ceutical acceptable salt thereof; i) Peak plasma concentrations of 2,4-dinitrophenol (C max ) steady state; ii) an average half-life (t 1 / 2 ); iii) The peak plasma concentration of 2,4-dinitrophenol (T max ) median time to reach; iv) Area under the curve (AUC) extrapolated to infinity for 2,4-dinitrophenol from about 3 h*μg / mL to about 420 h*μg / mLinf ) median; and v) AUC / C of about 18 max ratio To provide a method for achieving at least one of the above.

[0159] In some embodiments, the disclosure provides a method of treating a mitochondrial-related disorder or condition in a subject without causing a clinically significant risk of an adverse event, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0160] In some embodiments, the disclosure provides a method for reducing toxicity or side effects when treating a mitochondrial-related disorder or condition in a subject, the method comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0161] In some embodiments, the disclosure provides a method for reducing the toxicity or side effects of 2,4-dinitrophenol in treating a mitochondrial-related disorder or condition in a subject, the method comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0162] In some embodiments, the disclosure provides a method for preventing overdosing in treating a mitochondrial-related disorder or condition in a subject, the method comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0163] In some embodiments, the disclosure provides a method for preventing overdosage of 2,4-dinitrophenol in treating a mitochondrial-related disorder or condition in a subject, the method comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0164] In some embodiments, mitochondrial-related diseases include obesity, excess body fat, diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, cardiovascular disease, atherosclerosis, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, Rett syndrome, metabolic syndrome associated with aging, metabolic diseases associated with increased reactive oxygen species (ROS), Friedreich's ataxia, or liver disease.

[0165] In some embodiments, the disorder is a branched chain amino acid (BCAA) metabolism disorder, a lysosomal storage disorder, or a glycogen storage disorder.

[0166] In some embodiments, the diabetes is type 2 diabetes mellitus (T2DM).

[0167] In some embodiments, the cardiovascular disease comprises heart failure, HFpEF, HFrEF, HFmrEF, heart attack, coronary artery disease or CHD.

[0168] In some embodiments, the liver disease comprises non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), non-cirrhotic NASH, non-cirrhotic NASH with liver fibrosis, hepatic steatosis, liver fibrosis, cirrhosis, or hepatocellular carcinoma.

[0169] In some embodiments, the mitochondrial-associated disorder comprises cardiovascular disease, hypertension, type 2 diabetes, dyslipidemia, obesity, or nonalcoholic steatohepatitis (NASH).

[0170] In some embodiments, the mitochondrial associated condition is at least one of steatosis, inflammation, fibrosis, cirrhosis, and hepatocellular injury in NASH.

[0171] In some embodiments, the toxicity, adverse events, side effects and overdosage are associated with the mitochondrial uncoupling agent.

[0172] In some embodiments, the mitochondrial uncoupling agent is 2,4-dinitrophenol.

[0173] In some embodiments, the method comprises: i) The half-life of 2,4-dinitrophenol (t 1 / 2 ) to be extended; ii) Time to maximum plasma concentration of 2,4-dinitrophenol (T max ) to delay; iii) Peak plasma concentration of 2,4-dinitrophenol (C max ) ; and iv) Increasing the area under the curve (AUC) Includes at least one of the following:

[0174] In some embodiments, the disclosure provides a method for increasing metabolic rate without causing a clinically significant risk of an adverse event in a subject, the method comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0175] In some embodiments, the disclosure provides a method for increasing resting energy expenditure in a subject, the method comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0176] In some embodiments, the disclosure provides a method for treating a metabolic disorder in a subject, the method comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutically acceptable salt thereof.

[0177] In some embodiments, the subject suffers from at least one of obesity, excess body fat, type 2 diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, atherosclerosis, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, Rett syndrome, metabolic syndrome associated with aging, metabolic disease associated with increased reactive oxygen species (ROS), Friedreich's ataxia, NAFLD, NASH, non-sclerotic NASH, non-sclerotic NASH with liver fibrosis, hepatic steatosis, hepatic fibrosis, cirrhosis, and hepatocellular carcinoma.

[0178] In some embodiments, the methods include increasing resting metabolic rate without causing a clinically significant risk of an adverse event.

[0179] In some embodiments, the resting metabolic rate is increased by at least 10%.

[0180] In some embodiments, the resting metabolic rate is increased by at least 20%.

[0181] In some embodiments, the subject experiences an increase in resting energy expenditure of at least 10% following administration.

[0182] In some embodiments, the subject experiences an increase in resting energy expenditure of at least 20% following administration.

[0183] In some embodiments, the subject experiences an increase in resting energy expenditure of about 30% following administration.

[0184] In some embodiments, the method slows the progression of at least one of atherosclerosis, NAFLD, NASH, non-sclerotic NASH, non-sclerotic NASH with liver fibrosis, hepatic steatosis, hepatic fibrosis, cirrhosis, and hepatocellular carcinoma.

[0185] In some embodiments, the method enhances the body's natural processes and improves cardiac metabolic processes.

[0186] In some embodiments, the disclosure provides a method for treating hypertriglyceridemia in a subject associated with cardiovascular disease, atherosclerosis, obesity, hypertension, diabetes, insulin resistance, and / or liver disease, comprising administering a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof, to the subject.

[0187] In some embodiments, the subject has moderate hypertriglyceridemia associated with cardiovascular disease, atherosclerosis, obesity, hypertension, diabetes, insulin resistance, and / or liver disease; or severe hypertriglyceridemia associated with cardiovascular disease, atherosclerosis, obesity, hypertension, diabetes, insulin resistance, and / or liver disease.

[0188] In some embodiments, the disclosure provides a method for treating severe hypertriglyceridemia in a subject, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0189] In some embodiments, the subject has a triglyceride blood level greater than 500 mg / dL.

[0190] In some embodiments, the subject has severe hypertriglyceridemia associated with cardiovascular disease, atherosclerosis, obesity, hypertension, diabetes, insulin resistance, and / or liver disease.

[0191] In some embodiments, the subject has treatment-resistant hypertriglyceridemia.

[0192] In some embodiments, the subject has treatment-resistant severe hypertriglyceridemia.

[0193] In some embodiments, the subject has treatment-resistant severe hypertriglyceridemia associated with cardiovascular disease, atherosclerosis, obesity, hypertension, diabetes, insulin resistance, and / or liver disease.

[0194] In some embodiments, the subject suffers from at least one of abdominal pain, pain in the mid-abdominal, chest, or back area, gastrointestinal pain, difficulty breathing, loss of appetite, nausea, vomiting, inflammation of the pancreas, memory loss, dementia, xanthelasma, ringworm, and xanthomas.

[0195] In some embodiments, the subject is an adult male.

[0196] In some embodiments, the subject is of Hispanic descent.

[0197] In some embodiments, the methods include lowering low density lipoprotein cholesterol levels and / or lowering non-high density lipoprotein cholesterol levels.

[0198] In some embodiments, the method comprises: i) reducing triglyceride levels by at least 5%, at least 10%, or at least 20%; ii) reducing low density lipoprotein cholesterol levels by at least 5%, at least 10%, or at least 20%; and iii) reducing non-high density lipoprotein cholesterol levels by at least 5%, at least 10%, or at least 20%. Includes at least one of the following:

[0199] In some embodiments, the disclosure provides a method for reducing liver fat in a subject by at least 50%, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0200] In some embodiments, the disclosure provides a method for reducing lipids in a subject by at least 10%, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutically acceptable salt thereof.

[0201] In some embodiments, the disclosure provides a method of treating or reducing the risk of cancer in a subject, comprising administering to the subject a therapeutically effective amount of Compound 1, or a pharma- ceutical acceptable salt thereof.

[0202] In some embodiments, the cancer includes biliary tract cancer, bladder cancer, brain cancer (i.e., meningioma), breast cancer (postmenopausal), cervical cancer, colorectal cancer, endometrial / uterine cancer, esophageal cancer, gallbladder cancer, head and neck cancer, kidney / renal cancer, leukemia, liver cancer, multiple myeloma, non-Hodgkin's lymphoma, ovarian cancer, pancreatic cancer, gastric cancer, and thyroid cancer, and prostate cancer.

[0203] In some embodiments, the cancer is associated with obesity, excess body fat, diabetes, hypertension, dyslipidemia, metabolic disease, liver disease, and / or cardiovascular disease.

[0204] In some embodiments, the disclosure provides a method of treating obesity, cancer, excess body fat, type 2 diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, atherosclerosis, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, Rett syndrome, metabolic syndrome associated with aging, metabolic disease associated with increased reactive oxygen species (ROS), Friedreich's ataxia, NAFLD, NASH, non-sclerotic NASH, non-sclerotic NASH with liver fibrosis, hepatic steatosis, hepatic fibrosis, cirrhosis, or hepatocellular carcinoma, comprising administering a therapeutically effective amount of Compound 1 or a pharmacologic acceptable salt thereof to a subject; i) Peak plasma concentrations of 2,4-dinitrophenol (C max ) steady state; ii) a half-life (t) of 2,4-dinitrophenol of about 20 to 50 hours, about 25 to 40 hours, or about 30 to 40 hours. 1 / 2 ) median; iii) The peak plasma concentration of 2,4-dinitrophenol (T max ) median time to reach; iv) Area under the curve (AUC) extrapolated to infinity for 2,4-dinitrophenol from about 3 h*μg / mL to about 420 h*μg / mL inf ) median; and v) AUC / C of about 18 max ratio To provide a method for achieving at least one of the above.

[0205] In some embodiments, the method further includes determining the subject's Fibroscan® vibration-damped transient elastography (VCTE), Fibroscan® controlled attenuation parameter (CAP) score, magnetic resonance imaging proton density fat fraction (MRI-PDFF), and enhanced liver fibrosis (ELF) score before and after administration.

[0206] In some embodiments, the subject has a CAP score of greater than 300 dB / m prior to administration.

[0207] In some embodiments, the subject has liver fat by MRI-PDFF of at least 8% prior to administration.

[0208] In some embodiments, the subject has an elevated body mass index (BMI).

[0209] In some embodiments, the subject is at least about 28.0 kg / m 2 to about 45.0 kg / m 2 has a BMI of.

[0210] In some embodiments, the diabetes is type 2 diabetes mellitus (T2DM).

[0211] In some embodiments, the cardiovascular disease comprises heart failure, HFpEF, HFrEF, HFmrEF, heart attack, coronary artery disease or CHD.

[0212] In some embodiments, the liver disease comprises non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), non-cirrhotic NASH, non-cirrhotic NASH with liver fibrosis, hepatic steatosis, liver fibrosis, cirrhosis, or hepatocellular carcinoma.

[0213] In some embodiments, the condition associated with mitochondria is at least one of steatosis, inflammation, fibrosis, cirrhosis, and hepatocellular injury in NASH.

[0214] In some embodiments, the subject has non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), and / or hepatic steatosis.

[0215] In some embodiments, the subject suffers from type 2 diabetes, obesity, HFpEF, HFrEF, NAFLD, and / or NASH.

[0216] In some embodiments, the subject is suffering from liver inflammation, fibrosis, or cirrhosis.

[0217] In some embodiments, the subject experiences no significant systemic toxicity, no serious side effects, no clinically significant risk of adverse events, and / or no overdose following administration.

[0218] In some embodiments, the toxicity, adverse events and side effects are associated with a mitochondrial uncoupling agent.

[0219] In some embodiments, the mitochondrial uncoupling agent is 2,4-dinitrophenol.

[0220] In some embodiments, subjects do not experience any clinically significant risk of adverse events, side effects, toxicity, and / or overdosage associated with 2,4-dinitrophenol, such that subjects in need of treatment may be treated safely without serious side effects and risk of overdosage.

[0221] In some embodiments, the adverse events or side effects include at least one of nausea, vomiting, sweating, dizziness, headache, cataracts, glaucoma, fever, hyperthermia, tachycardia, hyperhidrosis, tachypnea, and death.

[0222] In some embodiments, the adverse events or side effects include at least one of fever, hyperthermia, tachycardia, hyperhidrosis, tachypnea, and death.

[0223] In some embodiments, the adverse event or side effect is characterized by at least one of elevated body temperature, elevated heart rate, excessive sweating, erythema, sweating, dehydration, and abnormal respiratory rate.

[0224] In some embodiments, the adverse event or side effect is associated with cardiovascular collapse, cardiac arrest, and / or death.

[0225] In some embodiments, the adverse event or side effect is associated with cardiac arrest.

[0226] In some embodiments, the subject does not experience a significant increase in body temperature or a significant increase in heart rate.

[0227] In some embodiments, subjects experience saturable absorption of Compound 1 such that overdosing is prevented. In some embodiments, there is saturation of absorption at high single doses. In some embodiments, there is saturation of absorption at single oral doses of Compound 1 of more than 500 mg, more than 600 mg, more than 700 mg, more than 800 mg, more than 900 mg, more than 1000 mg, more than 1050 mg, more than 1100 mg, more than 1200 mg, more than 1300 mg, or more than 1400 mg.

[0228] In some embodiments, the subject experiences no correlation between dose and toxicity, adverse events, side effects, or overdosing.

[0229] In some embodiments, the clinically significant risk of adverse events, side effects, toxicity, and / or overdose is determined by: i) The half-life of 2,4-dinitrophenol (t 1 / 2 ) to be extended; ii) Time to maximum plasma concentration of 2,4-dinitrophenol (T max ) to delay; iii) Peak plasma concentration of 2,4-dinitrophenol (C max ) ; and iv) Increasing the area under the curve (AUC) is prevented by at least one of the following:

[0230] In some embodiments, a clinically significant risk of adverse events, side effects, toxicity, and / or overdose is reduced by administration of Compound 1 at a peak plasma concentration (C max ) steady state.

[0231] In some embodiments, a clinically significant risk of adverse events, side effects, toxicity, and / or overdose is reduced by administration of Compound 1 within about 20-50 hours, about 25-40 hours, or about 30-40 hours of the mean half-life of 2,4-dinitrophenol (t 1 / 2 ) is provided.

[0232] In some embodiments, the clinically significant risk of adverse events, side effects, toxicity, and / or overdose is reduced by about 6-8 hours or about 6-10 hours of peak plasma concentration of 2,4-dinitrophenol (T max This is prevented by providing a median time to reach

[0233] In some embodiments, a clinically significant risk of adverse events, side effects, toxicity, and / or overdose is reduced by administration of Compound 1 at a median area under the curve extrapolated to infinity (AUC) of 2,4-dinitrophenol of about 3 h*μg / mL to about 420 h*μg / mL. inf ) is provided.

[0234] In some embodiments, a clinically significant risk of adverse events, side effects, toxicity, and / or overdose is reduced by administration of Compound 1 at an AUC / C of about 18. max This is prevented by providing a ratio.

[0235] In some embodiments, the various methods described above include: i) The maximum plasma concentration (T max ) median time to reach; ii) A half-life (t 1 / 2 ) median; and iii) Area under the curve (AUC) extrapolated to infinity for Compound 1 from about 18 h*ng / mL to about 380 h*ng / mL inf ) median The present invention includes providing at least one of the following:

[0236] In some embodiments, the subject experiences a decrease in at least one of weight, blood pressure, and blood glucose following administration.

[0237] In some embodiments, the subject i) weight loss of at least 5% or 10%; ii) a reduction in blood pressure of at least 5 mmHg; iii) HbA of at least 0.5% or at least 1.5% 1c Decrease in; iv) lipid reduction of at least 10%; v) at least 50% reduction in liver fat; vi) decreased serum alanine aminotransferase (ALT); and vii) Decreased Aspartate Aminotransferase (AST) experience at least one of the following:

[0238] compound 1 Compound 1 refers to 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole.

[0239] In some embodiments, Compound 1 is administered at about 30 mg, 100 mg, 200 mg, 500 mg, or 1050 mg per day.

[0240] In certain embodiments, the therapeutically effective amount is from about 30 mg to about 1400 mg per day, from about 50 mg to about 100 mg per day, from about 150 mg to about 600 mg per day, or from 200 mg to 550 mg per day.

[0241] In certain embodiments, the therapeutically effective amount is about 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg or 600 mg per day.

[0242] In certain embodiments, the therapeutically effective amount is about 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg or 95 mg per day.

[0243] In certain embodiments, the therapeutically effective amount is about 150 mg, 300 mg, or 450 mg per day.

[0244] In some embodiments, the therapeutically effective amount is selected to modulate Cmax, Tmax, and AUC.

[0245] Pharmaceutical Salts The compounds of formula I can be used as such or as salts. In cases where the formation of stable non-toxic acid or base salts is desired, administration of the compounds as pharma- ceutically acceptable salts may be appropriate.

[0246] Suitable pharma- ceutically acceptable salts include sulfate, bisulfate, hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, sulfuric acid, phosphoric acid, formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, gluconic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, glucuronic acid, maleic acid, fumaric acid, pyruvic acid, aspartic acid, glutamic acid, benzoic acid, anthranilic acid, 4-hydroxybenzoic acid, phenylacetic acid, mandelic acid, embonic acid (pamoic acid), methanesulfonic acid, ethanesulfonic acid, and the like. salts prepared from inorganic and organic acids, including carboxylic acid, benzenesulfonic acid, pantothenic acid, trifluoromethanesulfonic acid, sulfanilic acid, stearic acid, alginic acid, 2-hydroxyethanesulfonic acid, p-toluenesulfonic acid, cyclohexylaminosulfonic acid, salicylic acid, galactaric acid, β-hydroxybutyric acid, and galacturonic acid; or salts prepared from ammonium salts and metal salts, including calcium salts, magnesium salts, potassium salts, sodium salts, and zinc salts.

[0247] Composition / Formulation The pharmaceutical compositions of the present disclosure may be manufactured by processes well known in the art, e.g., by conventional mixing, dissolving, granulating, dragee-making, wet-commuting, emulsifying, encapsulating, entrapping, freeze-drying processes or spray-drying.

[0248] Pharmaceutical compositions for use according to the present disclosure may be formulated in a conventional manner using one or more pharma- ceutically acceptable carriers, including excipients and auxiliaries that facilitate the processing of active compounds into pharma- ceutically usable preparations. Appropriate formulations depend on the selected route of administration. Pharmaceutically acceptable excipients and carriers are generally known to those skilled in the art and are therefore included in the present disclosure. Such excipients and carriers are described, for example, in “Remington's Pharmaceutical Sciences” Mack Pub. Co., New Jersey (1991).

[0249] In some embodiments, the pharmaceutical composition comprises Compound 1, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable excipient.

[0250] In some embodiments, the methods of the disclosure include administering to a subject a pharmaceutical composition comprising Compound 1, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable excipient.

[0251] Pharmaceutical dosage forms The present disclosure includes novel pharmaceutical dosage forms of Compound 1 or a pharma- ceutically acceptable salt thereof. The dosage forms described herein are suitable for oral administration to a subject. The dosage form may be any form suitable for oral administration, including, but not limited to, a capsule or tablet. In some embodiments, the present disclosure provides a single unit dose capsule or tablet dosage form containing about 30 mg to about 1400 mg, about 100 mg to about 1000 mg, about 150 mg to about 600 mg, or about 200 mg to 550 mg of Compound 1 or a pharma- ceutically acceptable salt thereof. In some embodiments, Compound 1 is administered in a hydroxypropyl methylcellulose capsule.

[0252] In some embodiments, the amount of Compound 1 in the unit dose is about 30mg, 50mg, 75mg, 100mg, 150mg, 170mg, 200mg, 250mg, 300mg, 340mg, 350mg, 400mg, 450mg, 500mg, 510mg, 550mg, 600mg, 650mg, 700mg, 750mg, 800mg, 850mg, 900mg, 950mg, 1000mg, 1050mg, 1100mg, 1150mg, 1200mg, 1250mg, 1300mg, 1350mg or 1400mg.In some embodiments, the single unit dosage form is a capsule.In some embodiments, the single unit dosage form is a tablet.

[0253] In some embodiments, the amount of Compound 1 in the unit dose is about 30mg, 40mg, 50mg, 60mg, 70mg, 75mg, 80mg, 90mg, 100mg, 150mg, 170mg, 200mg, 250mg, 300mg, 340mg, 350mg, 400mg, 450mg, 500mg, 510mg, 550mg, 600mg, 650mg, 700mg, 750mg, 800mg, 850mg, 900mg, 950mg, 1000mg, 1050mg, 1100mg, 1150mg, 1200mg, 1250mg, 1300mg, 1350mg or 1400mg.In some embodiments, the single unit dosage form is a capsule.In some embodiments, the single unit dosage form is a tablet.

[0254] In some embodiments, the amount of Compound 1 in a unit dose is about 30mg, 100mg, 200mg, 500mg, 600mg, 1050mg or 1400mg. In some embodiments, the amount of Compound 1 in a unit dose is about 200mg, 400mg or 550mg. In some embodiments, the amount of Compound 1 in a unit dose is about 170mg, 340mg, 510mg. In some embodiments, the amount of Compound 1 in a unit dose is about 150mg, 300mg, 450mg.

[0255] In some embodiments, the amount of Compound 1 in the unit dose is about 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg or 95 mg per day.

[0256] Route of administration In therapeutic use for controlling or preventing weight gain in a mammal, the compounds of the present disclosure or pharmaceutical compositions thereof may be administered orally or parenterally.

[0257] In certain embodiments, a compound of the present disclosure or a pharmaceutical composition thereof may be administered orally once daily. EXAMPLES

[0258] [Table 16-1] [Table 16-2] Example 1: Preclinical studies

[0259] The therapeutic activity of Compound 1 was evaluated by oral administration in a mouse model of NASH (DIAMOND™ mice) and a rat model of metabolic syndrome (Zucker diabetic fatty rats). Compound 1 showed efficacy in these models at dose levels of 5 mg / kg / day and above in mice and 0.5 mg / kg / day and above in rats.

[0260] The PK of compound 1 was evaluated in mice, rats, and dogs. Compound 1 was converted to 2,4-dinitrophenol and rapidly absorbed in all animal species. The results showed that the time to peak plasma concentration of 2,4-dinitrophenol (T max ) was significantly delayed. Figure 1 shows the plasma concentrations of 2,4-dinitrophenol after administration of Compound 1 and 2,4-dinitrophenol.

[0261] Administration of Compound 1 also resulted in a significantly higher AUC / C of 2,4-dinitrophenol compared with administration of 2,4-dinitrophenol directly. max This optimized pharmacokinetic profile of 2,4-dinitrophenol following oral administration of compound 1 led to a significant improvement in the tolerability and safety evaluation in animals. Furthermore, the in vitro plasma protein binding of compound 1 was tested in many animal species, and the tissue distribution and excretion following oral administration to rats were investigated. The metabolism of compound 1 was evaluated in vitro using liver microsomes and hepatocytes from both nonclinical species and humans, as well as using human recombinant metabolic enzymes. These results led to the selection of rats and dogs as rodents and nonrodents for the pivotal safety studies. Preliminary investigations into the potential drug-drug interactions of compound 1 revealed no potential drug-drug interactions.

[0262] Compound 1 was evaluated in a nonclinical toxicology program, including acute toxicity / tolerability studies, repeated-dose toxicity studies, and genotoxicity studies, in accordance with the International Council for Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) Guidance M3(R2). All pivotal studies supporting the safety evaluation were conducted in accordance with the U.S. Food and Drug Administration (FDA) Good Laboratory Practice (GLP). Overall, results from single-dose tolerability and repeated-dose toxicity studies in mice, rats, and dogs indicate that compound 1 was well tolerated at doses up to 100 mg / kg / day in mice (for 7 days), 120 mg / kg / day in rats (for 63 days), and 100 mg / kg / day in dogs (for 61 days).

[0263] There were no compound 1-related deaths or effects on body weight, body weight gain, food intake, temperature, ophthalmology, electrocardiogram, hematology, clinical chemistry or urinalysis (UA).

[0264] Example 2: Single Ascending Dose (SAD) Study

[0265] A double-blind, placebo-controlled Phase I study of the safety and pharmacokinetics of single ascending doses of Compound 1 was conducted in healthy volunteers. Compound 1 was administered orally to volunteers at doses ranging from 30 mg once daily (QD) to 1400 mg QD in a total of 67 subjects (50 active / 17 placebo) across eight cohorts. The objectives of the study are to: Establish the plasma pharmacokinetic profiles of compound 1 and 2,4-dinitrophenol following single doses in healthy volunteers. Establish dose / exposure relationships for a single dose of Compound 1 across a wide range of exposures in healthy volunteers. · Estimate the pharmacokinetic effects of a 50% fat meal in combination with Compound 1. Characterize the pharmacokinetics of compound 1 and 2,4-dinitrophenol.

[0266] The screening process took place 28 days prior to admission and lasted 3 days. In all cohorts except cohort 4 (fed / fasted), subjects were admitted to the clinical research unit for up to 4 days. Administration of a single dose of the study drug in the fasting state (8-h fast) was performed on day 1, except for the crossover food effect cohort, in which a 10-h fast was performed before eating. After all safety assessments and blood sampling for PK analysis were completed, subjects were discharged on day 3. The fed / fasted cohorts remained hospitalized and were discharged on day 7.

[0267] Double-blind dosing was performed in cohorts 1 to 8. In these cohorts, 6 subjects received Compound 1 and 2 received matching placebo. Dose escalation was in the following order: 30 mg, 100 mg, 200 mg, 500 mg, 1400 mg, 1050 mg, and 600 mg. Cohort 4 was transitioned to fed state (50% fat diet) dosing after pharmacokinetic testing and had a 72-hour hospital washout after the first dose before the final dose.

[0268] Dose escalation is shown in Table 1 below. [Table 1]

[0269] Subjects were admitted to the clinical research unit on day 1. Dosing occurred on the morning of day 1 after an 8-h fast, except for the fed cohort, which had a 10-h fast before the meal. Blood samples for PK parameter evaluation were taken according to the evaluation schedule. ICs were taken according to the evaluation schedule. Subjects were discharged on day 3 after completion of all blood samples and safety evaluations. The fed / fasted cohorts remained hospitalized and were discharged on day 7.

[0270] Schematic diagram of study design [Table 17]

[0271] Drug administration

[0272] Compound 1 or matching placebo was administered orally as a single dose. All subjects, except for the fed cohort, were dosed in the morning after an 8-hour fast, remained in a semi-reclined position for 1 hour, and fasted for 4 hours after dosing. Capsules were swallowed with 240 mL (8 fl oz) of room temperature water.

[0273] Cohort 4 was a crossover food-effect analysis. Subjects enrolled in Cohort 4 were administered 30 minutes after the start of a standardized (containing 50% fat) meal (after a 10-hour fast) to assess the effect of food. Study medication was administered with 240 mL (8 fluid ounces) of room temperature water.

[0274] This study shows that the 2,4-dinitrophenol curve is extended in all cohorts. Compound 1 maintains the efficacy of 2,4-dinitrophenol while increasing C max Flattening the curve reduces toxicity and provides a "trickle-drip" effect for near 24 / 7 delivery.

[0275] 2A and 2B show the AUC of 2,4-dinitrophenol after administration of Compound 1. It is shown that 2,4-dinitrophenol is absorbed saturably after administration of Compound 1, and excessive intake of 2,4-dinitrophenol can be prevented.

[0276] Pharmacokinetics of Compound 1 and 2,4-Dinitrophenol

[0277] Compound 1 was administered in fasted state at 30 mg, 100 mg, 200 mg, 500 mg and 1050 mg.

[0278] Compound 1 is rapidly absorbed and max The median time was 1.75 to 3.00 hours, and T lag The median t was less than 0.50 hours at all dose levels. 1 / 2 The time to clearance was short, ranging from 1.01 to 2.32 hours in the 500 mg and 1050 mg fasting cohorts. The mean apparent clearance and volume of distribution appeared to increase with increasing dose. Dose-normalized C max Compound 1 exposure based on AUC increased dose-proportionally between the 30 mg and 200 mg dose groups and did not appear to be dose-proportional at doses above 200 mg (500 mg and 1050 mg).

[0279] 2,4-Dinitrophenol was rapidly observed after administration of Compound 1, and T lag The median time was less than 0.50 hours, and T max The median t ranged from 6.01 to 10 hours. 1 / 2 The mean apparent clearance and volume of distribution were higher in the 500 mg and 1050 mg dose groups compared with the 30 mg, 100 mg, or 200 mg dose groups. max 2,4-dinitrophenol exposure based on and AUC appeared to increase less than dose proportionally between the 30 mg, 100 mg, or 200 mg dose levels and the 500 mg or 1050 mg dose levels, with less than an 18-fold increase in exposure with a 35-fold dose increase.

[0280] Example 3: Evaluation of food effects

[0281] A single 500 mg dose of Compound 1 capsule was taken in the fasted state followed by a single 500 mg dose of Compound 1 capsule with a standardized (containing 50% high fat) meal (after a 10 hour fast). max The geometric means of T were 14.8 ng / mL and 25.3 ng / mL, with geometric mean CV% of 52.1% and 49.3%, respectively, which showed moderate variability. max The median AUC was 3.0 hours under fed conditions and 6.0 hours under fed conditions. 0-24h The geometric mean AUCs were 53.5h*ng / mL and 273h*ng / mL, and the geometric mean CV% was 86.5% and 7.92%, respectively, showing high to low variability. last The geometric mean CV% for the fasted and fed groups was 66.2% and 44.3%, respectively, with moderate variability.

[0282] The geometric mean ratio (90% CI) of compound 1 fed (test) vs fasted (reference) was C max 1.82 (1.30-2.56), AUC 0-24h 5.11 (1.75-14.9), and AUC last The result was 4.19 (2.75-6.39).

[0283] 2,4-Dinitrophenol C max The geometric means of T were 694 ng / mL and 1680 ng / mL, with moderate variability of geometric mean CV% of 31.6% and 23.3%, respectively. max The median AUC was 8.0 hours under fed conditions and 18.0 hours under fed conditions. 0-24h The geometric mean AUCs were 12400 h*ng / mL and 25300 h*ng / mL, with moderate to low variability, with geometric mean CV% of 31.3% and 18.0%, respectively. lastwere 34,200 h*ng / mL and 94,000 h*ng / mL, and the geometric mean CV% for the fasted and fed groups was 46.2% and 25.9%, respectively, indicating moderate to low variability.

[0284] The geometric mean ratio (90% CI) of 2,4-dinitrophenol fed (test) vs fasted (reference) was C max 2.35 (1.85-2.99), AUC 0-24h 2.03 (1.53-2.70), and AUC last The result was 2.58 (1.88-3.56).

[0285] Figures 3a and 3b show the comparison of plasma Compound 1 concentrations (linear and semi-logarithmic scales) in different dietary states after oral administration of 500 mg of Compound 1. Figures 4a and 4b below show the comparison of plasma 2,4-dinitrophenol concentrations (linear and semi-logarithmic scales) in different dietary states after oral administration of 500 mg of Compound 1.

[0286] A summary of the plasma pharmacokinetic parameters after administration of Compound 1 is shown in Table 2 below. A summary of the plasma pharmacokinetic parameters of 2,4-dinitrophenol after administration of Compound 1 is shown in Table 3 below. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6]

[0287] In the completed SAD study, Compound 1 was rapidly absorbed and converted to saturable levels of 2,4-dinitrophenol with a mean half-life of 1-3 hours. 2,4-dinitrophenol levels were significantly higher with a mean half-life of less than 40 hours. A positive effect of food on Compound 1 absorption was evident, as was a positive effect of Compound 1 particle size on absorption, as two Compound 1 formulations with different particle sizes were evaluated. There was evidence of saturable absorption at high single doses. AUC / C max The ratio was approximately 18 regardless of dose.

[0288] Single oral doses of 30-1400 mg of Compound 1 were found to be safe and well tolerated in all subjects in the SAD study. No serious adverse events (SAEs) were reported. No empirical correlation was found between dose and incidence of adverse events. The most representative system organ class (SOC) of adverse events (AEs) was gastrointestinal, mainly lower abdominal discomfort and loose stools / diarrhea, with a higher incidence observed in the Compound 1 treatment group than in the placebo group. All AEs were mild or moderate in intensity, with the majority being judged as mild. Evaluation of ECG, vital signs, and clinical laboratory values ​​showed no trend for abnormal findings or association with dose. Green discoloration of urine observed in some subjects at higher doses appears to be a harmless phenomenon.

[0289] SAD research has shown that: The pharmacokinetics of Compound 1 is characterized by rapid absorption and rapid excretion. 2,4-Dinitrophenol appears rapidly and is excreted relatively slowly. This metabolite is rapidly formed, excreted relatively slowly, and has low circulating levels (<1%) compared to 2,4-dinitrophenol. Administration of a standard high fat (50%) meal delayed the absorption of Compound 1 and the appearance of 2,4-dinitrophenol while increasing the total exposure (Cmax and AUC) of Compound 1 and 2,4-dinitrophenol. Compared to fasted conditions, under fed conditions, the Cmax of Compound 1 and 2,4-dinitrophenol increased by more than 1.8-fold, and the AUC of Compound 1 and 2,4-dinitrophenol increased by more than 4.0-fold and more than 2.0-fold, respectively.

[0290] Example 4: Multiple Ascending Dose (MAD) Study

[0291] A double-blind, sponsored, open-label, placebo-controlled Phase I study was conducted in volunteers with high MBI to evaluate the safety, pharmacokinetics, and pharmacodynamics of multiple ascending doses of Compound 1. The first cohort, including 10 healthy, high body mass index (BMI) subjects, completed a dose of 200 mg of Compound 1. Two further cohorts are planned at doses of 400 mg and 550 mg QD.

[0292] Subjects tolerated oral Compound 1 well at a dose of 200 mg QD. There were no SAEs, and preliminary evaluation of the data revealed no significant changes in physical examination, vital signs, ECG, or clinical laboratory data. Figure 6 also shows that plasma concentrations of 2,4-dinitrophenol reached steady state and that the curves for 2,4-dinitrophenol were flat in all cohorts.

[0293] FIG. 6 shows that administration of Compound 1 did not result in a significant increase in body temperature.

[0294] Figure 7 below shows that Compound 1 increases resting energy expenditure (REE) by approximately 29% compared to placebo.

[0295] Figures 8 and 9 show that Compound 1 reduces body weight and glucose levels after administration.

[0296] Figures 10 and 11 show that Compound 1 reduces systolic and diastolic blood pressure after administration, which leads to significant risk reduction in patients with cardiovascular diseases such as HFpFF, HFrEF and HFmrEF. The results show that the reduction in blood pressure was statistically highly significant and changes were observed at all three dose levels. An almost immediate and rapid effect was observed on both diastolic and systolic blood pressure. The reduction in blood pressure, together with the reduction in blood glucose and fat fraction, strongly indicates that Compound 1 is effective in HFpEF. Figure 12 also shows that the reduction in blood pressure and blood glucose as well as the reduction in fat fraction are not accompanied by an increase in heart rate.

[0297] Example 5: Phase 2a study of Compound 1 in patients with elevated liver fat and high BMI

[0298] High liver fat and high body mass index (28-45 kg / m 2 A 61-day randomized, double-blind, placebo-controlled study evaluating the safety and efficacy of three doses of Compound 1 in patients with glaucoma or glaucoma is ongoing.

[0299] Research rationale

[0300] Subjects with high BMI and elevated liver fat better recapitulate the metabolic profile of the definitive patient population with NASH (Golabi et al., 2020). Given the importance of metabolic abnormalities in the pathogenesis of NASH, understanding the safety and therapeutic effects of compound 1 primarily on liver fat and secondarily on body weight in volunteers with high BMI and elevated liver fat will provide greater confidence in selecting a dose that may be effective for a Phase 2b clinical study. Establishing the dose and associated pharmacokinetic exposure that results in a significant reduction in liver fat (≥50% reduction in relative liver fat) in a 61-day study is expected to provide safety, target engagement, and efficacy data for a longer-term (9-month) Phase 2b study in biopsy-verified patients with NASH. Additionally, the clinical effects of mitochondrial uncoupling have been studied in populations reflecting similar obese metabolic profiles (Tainter et al., 1934; Harper et al., 2001), providing further historical precedent regarding safety for the patient population under study.

[0301] Overall, understanding the safety and efficacy of Compound 1 on liver fat and body weight in a subject population with the metabolic characteristics of the NASH patient population (high BMI and high liver fat) will provide greater confidence in dose selection and efficacy in subsequent long-term Phase 2b clinical studies.

[0302] Rationale for selected dose

[0303] Single exposures of all compounds 1 and 2,4-dinitrophenol showed acceptable AE profiles in the SAD study. Compound 1 has a short half-life, whereas 2,4-dinitrophenol has a half-life of approximately 40 hours. Significant accumulation of 2,4-dinitrophenol is expected at steady state when administered once daily. In cohort 1 of the MAD study, administration of 200 mg QD for 14 days resulted in a dose accumulation factor of approximately 3.5-fold, with steady state being reached by 7 consecutive days of daily dosing.

[0304] The pharmacodynamic effects of a single dose of Compound 1 in the SAD study were monitored using indirect calorimetry (IC) to ascertain changes in resting metabolic rate (RMR). Single dose exposure to Compound 1 at a high dose of 1400 mg in the SAD study resulted in an increase in RMR of up to approximately 20% through the first 33 hours of IC assessment post-dose.

[0305] Considering the safety of single dose exposure and the associated pharmacological effects on RMR, a starting repeated oral dose of 200 mg per day was expected to increase RMR by approximately 10%. This increase in RMR was confirmed in cohort 1 of the MAD study, where an increase in RMR of approximately 10% was observed by day 7 of oral dosing of 200 mg per day, which was sustained through day 15 of repeated dosing. Steady-state exposure of the 200 mg QD dose was as expected considering dose accumulation. Given the acceptable safety profile after 2 weeks of dosing with the 200 mg QD dose, the next cohort of the MAD study will receive compound 1 at 400 mg QD for 14 days. The expected increase in RMR is 20%, and steady-state dose exposure is expected to be within the range of the highest exposure achieved in the SAD study. If the 400 mg QD per day dose demonstrates an acceptable safety profile after 2 weeks of dosing, it is anticipated that subsequent cohorts will receive 550 mg QD of compound 1 per day, but this decision will depend on new data being reviewed in real time. The expected increase in RMR at this dose / exposure is 30%, which is within the range of dietary effects on RMR and below the acceptable safety profile seen in previous clinical studies of 2,4-dinitrophenol. Given that the half-life of 2,4-dinitrophenol is approximately 40 hours, it is likely that the steady-state exposure of 550 mg QD of compound 1 will be higher than the single-dose exposure measured in the SAD study. Nevertheless, population pharmacokinetic (PPK) modeling of compound 1 and 2,4-dinitrophenol concentrations indicates that the steady-state 2,4-dinitrophenol concentrations achieved with a dose of 550 mg of compound 1 per day will be below the levels observed in clinical and nonclinical studies that affect body temperature. Historically, 2,4-dinitrophenol has been associated with deleterious increases in body temperature at concentrations above 28,000 ng / mL in humans (Zhao 2015), and studies of compound 1 in dogs with plasma 2,4-dinitrophenol concentrations above 13,000 ng / mL have shown increased body temperature, panting, and erythema. Furthermore, an increase in RMR of approximately 200% would be expected to increase body temperature (Bachynsky 2015[US20150056160A1]).Therefore, the expected C when compound 1 was administered at 550 mg QD. max The increase in steady-state 2,4-dinitrophenol concentration and the resulting RMR are associated with an increase in body temperature. max and well below the increase in RMR.

[0306] The study duration of 61 days was chosen based on the dosing duration of the completed toxicity study of compound 1 and to reflect the clinical experience with 2,4-dinitrophenol from studies conducted by Maurice Tainter and Samuel Simkins (Tainter et al., 1934; Harper et al., 2001; Geisler, 2019). In these studies, 2,4-dinitrophenol was administered once daily for 1-3 months at doses that readily caused a 20-40% increase in RMR. The resulting experienced weight loss was 1.4-2.1 lbs / week. The drug was well tolerated at these dose levels for 1-3 months. At 61 days of dosing, compound 1 is expected to induce significant hepatic fat reduction in response to a reduction in body weight and a dose-related increase in RMR of 10-40%. For example, 150 mg of Compound 1 increases resting energy expenditure by 10%, 300 mg of Compound 1 increases resting energy expenditure by 20%, and 450 mg of Compound 1 increases resting energy expenditure by 30%. The resulting 61-day safety and efficacy data should provide important guidance for dose selection in longer-term Phase 2b trials.

[0307] primary purpose Efficacy: To evaluate the reduction in liver fat content, as assessed by magnetic resonance imaging proton density fat fraction (MRI-PDFF), from baseline to day 61 in subjects with a high BMI receiving Compound 1 compared to placebo. Safety: Evaluate the safety and tolerability of repeated daily doses of Compound 1 for 61 days in subjects with overweight and obesity as defined by BMI.

[0308] secondary purpose · Evaluate the rate and amount of weight loss following 61 days of Compound 1 treatment. Change from baseline in total body fat fraction after 61 days of Compound 1 treatment will be assessed by MRI. Characterize the PK profiles of compound 1 and 2,4-dinitrophenol over a 61-day treatment period in subjects with high BMI. Evaluate and correlate changes from baseline in liver composition measurements and changes in liver fat content following Compound 1 administration. · To investigate the pharmacodynamic (PD) effects of compound 1 on metabolic and cardiovascular risk factors. · To investigate the PD effect of compound 1 on metabolomic, proteomic and lipidomic profiles. Where data permit, determine the dose / exposure relationship of efficacy and PD effects of Compound 1.

[0309] Endpoints

[0310] Primary endpoint Relative change from baseline in liver fat content by MRI-PDFF at day 61.

[0311] Secondary Endpoints Pharmacodynamics Change from baseline in body weight at day 61. Change from baseline in total body fat fraction at day 61. · Change from baseline in surrogate measures of liver inflammation and fibrosis at day 61: Fibroscan® Vibration-Containing Transient Elastography (VCTE), Fibroscan® Controlled Attenuation Parameter (CAP) score, and Extended Liver Fibrosis (ELF) score. Changes from baseline in lipid parameters and cardiovascular risk biomarkers at day 61: serum high-sensitivity C-reactive protein (hs-CRP), Lp(A), Apo B, low-density lipoprotein (LDL), high-density lipoprotein (HDL), total cholesterol, triglycerides and free fatty acids (FFA). Changes from baseline in metabolic disease parameters at day 61: homeostasis model assessment of insulin resistance (HOMA-IR), fasting plasma glucose, glycated albumin, HbA 1c . Safety and Tolerability Summary of physical examination findings during the study period. Adverse event (AE) assessment during the study. Assessment of vital sign parameters throughout the study. Parameters include resting systolic and diastolic blood pressure, resting heart rate, resting respiratory rate, and oral temperature. Assessment of body weight during the study. · Safety 12-lead ECG for the duration of the study. Evaluation of clinical laboratory values ​​(hematology, complete chemistry panel (including lipid panel, CPK, magnesium, liver function tests), and urinalysis (UA)) over 61 days of treatment. - Evaluation of ophthalmologic examinations, including slit lamp examinations, pre-treatment and 61 days post-treatment. Pharmacokinetics Population PK analysis of compound 1 and 2,4-dinitrophenol. The following PK parameters are estimated accordingly: max , T max , t 1 / 2 , T lag , AUC 0-t , AUC 0-∞ , CL / F, Vd / F, λ z Other PK parameters may be calculated if data permit and appropriate. Non-compartmental analysis of compound 1 and 2,4-dinitrophenol, where data permit. The following PK parameters will be calculated: max , AUC, and accumulation. Modeling the exposure-response relationships of Compound 1 and 2,4-dinitrophenol, as well as efficacy / pharmacodynamic endpoints, as appropriate.

[0312] Exploratory Endpoints Change from baseline in metabolomic and lipidomic profiles at day 61 (One Way Liver-[OWL] metabolomic and lipidomic assays). Change from baseline in proteomic profile (SomaScan®) at day 61. Change from baseline in serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) concentrations at day 61, when data permit.

[0313] Study design

[0314] This is a Phase 2a randomized, parallel-group, placebo-controlled, double-blind, multiple-dose study evaluating the safety and efficacy of three oral doses of Compound 1 compared to placebo over a 61-day period in subjects with high BMI and elevated liver fat, as shown in the following scheme: [Table 18]

[0315] Subjects will be screened for 45 days and eligibility will be determined based on specific medical history, physical examination, laboratory tests, and imaging evaluations. Due to testing schedules, multiple visits will likely be required to complete the screening. However, if all screening evaluations, including MRI, and treatment can be completed within 30 days of the first dose, a single screening visit will be acceptable.

[0316] Eligible patients will be randomly assigned to either the Compound 1 treatment group or a matching placebo control group and will receive the compound once daily (fasting) for a total of 61 days. Subjects will be assessed at frequent clinic visits throughout the 61-day treatment period, with a follow-up visit within 10-14 days after the end of treatment.

[0317] Subjects will be instructed to maintain the same diet and activity / exercise levels for the duration of the study as they did before participating in the study.

[0318] Dosing will be administered once daily under fasting conditions. Subjects will be randomized equally to one of four treatment arms (N=20 per arm): Group 1: Matched placebo administered orally once daily for 61 days. Group 2: Compound 1 was orally administered at 150 mg once daily for 61 days. Group 3: Compound 1 was orally administered at 300 mg once daily for 61 days. Group 4: Compound 1 was orally administered at 450 mg once daily for 61 days.

[0319] Randomization was based on HbA 1c Blocked and stratified by (normal range vs. 5.7% to 9.0%, inclusive).

[0320] Inclusion criteria To be eligible, subjects must meet all of the following inclusion criteria: 1. Adult male or female, aged 28-65 years (inclusive) at the time of informed consent, with a BMI of 28.0-45.0 kg / m 2 (inclusive). a. Female subjects of childbearing potential must be non-lactating, non-pregnant with a negative urine pregnancy test at screening, and agree to continue effective contraception for at least 4 weeks or continue barrier methods from 2 weeks prior to the first study drug dose until 30 days after the last study drug dose. b. Female subjects of childbearing potential must not donate eggs during the study and for at least 30 days after the last dose of investigational product. c. Female subjects of non-childbearing potential must be surgically sterilized (e.g., hysterectomy, bilateral tubal ligation, oophorectomy) or postmenopausal (no menses for >1 year and follicle-stimulating hormone (FSH) >40 U / L at screening). d. Non-vasectomized male subjects, and / or subjects who have had a vasectomy but have not had two negative sperm tests post-surgery, must agree to use acceptable methods of contraception from the time of the first dose of study drug through 30 days after the last dose of study drug and to not donate sperm during the study and for at least 30 days after the last dose of study drug. 2. Inclusion by investigator assessment of general medical symptoms and documented by medical history, physical examination, vital signs assessment, 12-lead ECG, clinical laboratory assessment, and general observations. a. Subjects must be taking a stable dose of medication for an underlying condition associated with obesity for at least 2 months prior to screening. b. Diabetic patients may be treated with metformin, DPP-4 inhibitors, or sulfonylureas, but must have been on a stable dose for at least 2 months prior to screening. c. For screening, certain laboratory values ​​may be outside the reference ranges if they are consistent with the eligible subject's underlying obesity or related metabolic dysfunction (e.g., dyslipidemia and hyperglycemia), so long as these abnormalities do not suggest an underlying disease that may affect the safety of the subject in the study or that may interfere with the evaluation of Compound 1 or affect the interpretation of the study results. d. Abnormalities or deviations outside the normal ranges of other assessments (laboratory tests, ECG, vital signs, physical exam) that the Investigator determines to be clinically significant may be repeated once at the discretion of the Investigator(s). Results that continue to be outside the normal ranges must be determined by the Investigator to be not clinically significant and appropriate for study participation. e. Subjects with elevated unconjugated bilirubin due to presumed Gilbert syndrome are allowed. f. Subjects must be euthyroid as assessed at screening with a thyroid profile with thyroid stimulating hormone (TSH) and free thyroxine (T4) tests. Subjects with a stable history of thyroid disease and taking a stable dose of thyroid medication for at least 4 months may be enrolled. 3. Fibroscan® CAP score >300 dB / m. 4. Liver fat ≥ 8% by MRI-PDFF. 5. Understand the study procedures and requirements and provide written informed consent and authorization for disclosure of protected health information. 6. Willing and able to comply with the requirements of the clinical trial protocol.

[0321] Exclusion criteria Subjects will be excluded from the study if they meet any of the following criteria: 1. Insulin-controlled diabetes. 2. Pregnant, breastfeeding, or planning to become pregnant. 3. Subjects who cannot tolerate Magnetic Resonance Imaging (MRI) or cannot fit into an MRI scanner, or have any condition that contraindicates MRI testing, including but not limited to surgical clips / metal implants / shrapnel. Subjects must not be claustrophobic, have a history of claustrophobia, or have intolerance to closed or small spaces. 4. Weight gain or loss of >5% in the 3 months prior to the study or >10% in the 6 months prior to screening. 5. History of lap-band, intragastric balloon, duodenal-jejunal sleeve, or bariatric surgery within 5 years of screening, planned bariatric surgery before the end of study participation, or planned to lose weight through a special diet, exercise program, or both during the study. 6. History of malignant hyperthermia. 7. History of chronic severe recurrent skin rash of unknown cause. 8. History or ongoing clinically significant cardiovascular disease, including but not limited to transient ischemic attack, stroke, arrhythmia, syncope, unstable angina, myocardial infarction within 6 months prior to screening, congestive heart failure, or uncontrolled hypertension (defined as a systolic blood pressure ≥ 160mmHg or a diastolic blood pressure ≥ 100mmHg measured by the average of three resting blood pressure measurements in the seated position using an appropriately sized blood pressure cuff). 9. Resting heart rate <45 bpm or >110 bpm. 10. Screening ECG or medical history shows: Marked baseline prolongation of the QT / QTcF interval (e.g., repeated QTcF intervals >450 msec in men and >470 msec in women). b. Personal history of additional risk factors for Torsades de Pointes (TdP) (e.g., heart failure, hypokalemia, family history of long QT syndrome) or family history of sudden cardiac death of unknown cause. 11. Kidney disease, kidney transplant, or estimated glomerular filtration rate (eGFR) < 50 mL / min / 1.73 m based on the CKD-EPI creatinine equation (NKF 2009; https: / / www.kidney.org / content / ckd-epi-creatinine-equation-2009) 2 . 12. Significant lung disease requiring chronic daily medication, including chronic obstructive pulmonary disease (COPD), emphysema, pulmonary fibrosis or asthma. 13. Untreated Obesity Hypoventilation Syndrome (OHS) or Obstructive Sleep Apnea (OSA). 14. History of or ongoing (acute or chronic) liver disease other than nonalcoholic fatty liver disease (NAFLD) / nonalcoholic steatohepatitis (NASH), including but not limited to autoimmune liver disease, viral hepatitis, hereditary hemochromatosis, primary biliary cirrhosis, Wilson's disease, alpha-1-antitrypsin deficiency, alcoholic liver disease, acute fatty liver of pregnancy, or drug-induced (including acetaminophen) liver disease. 15. Clinically significant gastroparesis, inflammatory bowel disease, or history of any upper gastrointestinal surgery or treatment (except cholecystectomy and minor gastric surgery approved by a medical monitor). 16. History of cirrhosis and / or liver failure including ascites, hepatic encephalopathy or variceal bleeding. 17. History of acute pancreatitis within 1 year of screening or chronic pancreatitis from any cause. Serum triglyceride concentration greater than 18,500 mg / dL. 19.HbA 1c >9.0%. 20. Family member (mother / father / sibling) and / or personal history of retinal detachment at any time in the past. 21. Any history or current diagnosis of glaucoma. 22. Evidence at screening eye examination of: A peripheral retinal lesion or retinal break requiring treatment, or a retinal lattice requiring treatment. b. Diabetic retinopathy with macular exudate or edema as seen by optical coherence tomography (OCT) and laboratory. c. Progressive macular disease affecting vision, including macular folds (epidermal membranes) and macular degeneration. d. Visually significant cataract as determined by an ophthalmologist. e. History of any intravitreal injection of anti-VEGF agents for macular degeneration. f. History of vitreous surgery. 23. History of malignant neoplasms within 5 years of screening, except for basal or squamous cell skin cancer, cervical intraepithelial neoplasia, or prostate cancer that does not currently require or is not believed to require radiation therapy, chemotherapy, and / or surgical or hormonal intervention. 24. History of organ transplantation. 25. Received a COVID-19 vaccine within 1 week prior to dosing (Visit 2 / Day 1) and / or plans to receive a COVID-19 vaccine during the study period. 26. History of significant substance abuse within 1 year prior to screening, or heavy use of soft drugs (e.g., marijuana) within 3 months prior to the screening visit, or heavy use of hard drugs (e.g., cocaine, phencyclidine [PCP], opioid derivatives including heroin, and amphetamine derivatives) within 1 year prior to screening. 27. History of alcoholism within the past 2 years or current evidence of excessive alcohol use as assessed by screening using the Alcohol Use Disorders Identification Test (AUDIT, Thompson 2018), and a history of regular alcohol intake within 6 months of screening as determined by the investigator to exceed approximately 14 drinks / week for men and approximately 7 drinks / week for women (1 drink = 4 ounces (120 mL) of wine or 12 ounces (360 mL) of beer or 1 ounce (30 mL) of hard liquor). 28. A positive urine drug screen for drugs of abuse or a positive blood test for phosphatidylethanolamine (PEth) >100 ng / mL at screening. If the PEth value is exclusionary, enrollment may be considered if the investigator and medical monitor agree that the subject's medical history is not consistent with alcohol abuse. 29. Currently regularly use e-cigarettes or smoke 5 or more cigarettes or their equivalent per week. Use of nicotine patches to quit smoking is permitted. 30. Positive hepatitis B surface antigen (HBsAg), hepatitis C virus antibody (HCV Ab), or human immunodeficiency virus (HIV1 / 2) antibody. 31. Neutropenia (defined as absolute neutrophil count 1000 / μL or less). 32. Serum AST or ALT >5x upper limit of normal (ULN) at screening (one repeat test within 7 days allowed at the investigator's discretion). 33. Total bilirubin >ULN, except as caused by Gilbert's syndrome or as considered normal variation in the absence of other clinically relevant liver disorders as approved by the medical monitor. 34. International normalized ratio (INR) ≥ 1.3 at screening if there is other evidence of possible significant liver dysfunction. 35. Participation in other clinical trials at the time of screening, or exposure to any investigational drug, including topical medications, within 30 days of screening, or within 5 half-lives if the half-life is known. 36. No tattoos or body piercings during the study. Any underlying physical or mental illness that, in the opinion of the investigator or sponsor, would make the subject unlikely to be able to comply with the requirements of the study or to complete the study. 37. Any condition that, in the opinion of the investigator, may interfere with the ability to provide written informed consent or to comply with the instructions of the study, or that may confound the interpretation of the study results, or that may place the subject at undue risk. 38. Known or potential hypersensitivity to Compound 1 or any of its excipients. Prohibited drugs (currently in use): 39. Any herbal supplements, over-the-counter, mail-order or prescription drugs for weight loss. 40. Prescription or over-the-counter stimulants, including dextroamphetamine / Dexedrine, dextroamphetamine / amphetamine combinations / Adderall, or methylphenidate (Ritalin®, Concerta®). 41. Thiazolidinediones (TZDs): Pioglitazone / Actos, Rosiglitazone / Avandia. 42. Glucagon-like peptide 1 (GLP1) agonists: Exenatide / Byetta / Bydureon, lixisenatide / Adlyxin, liraglutide / Victoza, dulaglutide / Trulicity, semaglutide / Ozempic. 43. Sodium-glucose cotransporter-2 (SGLT2) inhibitors: canagliflozin / Invokana, dapagliflozin / Farxiga, empagliflozin / Jardiance, ertugliflozin / Steglatro. 44. Vitamin E: Use of ursodiol or high-dose vitamin E >400 IU / day for ≥1 month within the past 6 months or initiation of high-dose vitamin E within 3 months prior to screening. 45. Recent (within 3 months of screening) or current use of obeticholic acid / Ocaliva, systemic corticosteroids, methotrexate, tamoxifen, amiodarone, or long-term use of tetracyclines. 46. ​​Warfarin, heparin, factor Xa inhibitors (dabigatran, betrixaban, edoxaban, apixaban, and rivaroxaban). 47. Concomitant medications known to prolong the QT / QTc interval and be associated with an increased risk of torsades de pointes as identified in the listing category "Known Risk" on the https: / / crediblemeds.org / website. 48. Products containing cannabidiol (CBD).

[0322] Treatment Plans and Methods Study procedures must be performed as specified in the assessment schedule (Table 4). [Table 4-1] [Table 4-2] [Table 4-3]

[0323] Pharmacokinetic parameters The following PK parameters are determined from the concentration-time data: Population PK (PPK) analysis of compound 1 and 2,4-dinitrophenol. The following PK parameters are calculated: max , T max , t 1 / 2 , T lag , AUC 0-t , AUC 0-∞ , CL / F, Vd / F, λz. Other PK parameters may be calculated if data permit and appropriate. Non-compartmental analysis of compound 1 and 2,4-dinitrophenol, where data permit. The following PK parameters will be calculated: max , AUC, and accumulation.

[0324] Magnetic resonance imaging proton density fat fraction (MRI-PDFF)

[0325] Magnetic resonance imaging proton density fat fraction (MRI-PDFF) is a non-invasive, quantitative biomarker to assess liver fat content (steatosis). The percentage of fat in the liver or proton density fat fraction (PDFF) is measured using MR:MRI-PDFF at baseline and end of treatment. Liver volume is assessed from axial T1-weighted or dual-echo gradient-echo images covering the entire liver. This advanced MRI technique measures the fraction of mobile protons in the liver that are attributable to hepatic fat (PDFF), which is a direct indicator of liver fat content and a fundamental property of the tissue. Subjects are required to fast for 4 hours before MRI-PDFF is performed.

[0326] abdominal MRI

[0327] An MRI scan of the abdomen is taken to assess total fat. The two main measurements of this test are an estimate of total visceral adipose tissue (VAT), the type of fat stored within the body cavities, and subcutaneous adipose tissue (SAT), the type of fat visible just under the skin.

[0328] Fibroscan®

[0329] Fibroscan® is a non-invasive medical device that estimates the amount of liver fat (steatosis) and stiffness (fibrosis) of the liver. The test works by measuring the shear wave velocity. A small transducer at the end of an ultrasound probe transmits 50MHz waves to the liver. The tip of the probe also has a transducer that measures the speed of the shear wave (in meters per second) as it passes through the liver. The shear wave velocity is converted to liver stiffness, expressed in kilopascals (VCTE score). A second measurement is also taken to estimate liver steatosis by measuring the ultrasonic attenuation of the echo waves, called the Controlled Attenuation Parameter (CAP). Subjects are required to fast for 4 hours before undergoing Fibroscan®.

[0330] One Way Liver Assay (OWL)

[0331] OWL-Metabolomics

[0332] In this metabolomic assay, metabolites are extracted from the plasma and serum of volunteers to obtain a snapshot of cellular function. Liquid chromatography-mass spectrometry (LC-MS) metabolomics is used to identify serum biomarkers that distinguish between normal liver and NAFLD, and between NASH and NAFLD. The metabolomic profile also provides insight into cellular function and inflammation through the examination of various cellular metabolites that provide insight into key molecular pathways.

[0333] OWL-Lipidomics

[0334] Lipidomics is a non-invasive blood testing method to analyze and identify lipids in plasma and serum. These lipids are separated and characterized by mass spectrometry. Analysis includes fatty acids, fatty acid derivatives, glycerolipids, glycerophospholipids, sphingolipids and sterols.

[0335] Slow off-rate modified aptamer (SomaScan)

[0336] SomaScan is a non-invasive blood assay. Plasma and serum samples are probed with oligonucleotide aptamers whose three-dimensional conformation specifically binds to protein targets of interest. Over 7,000 proteins are probed and quantified, allowing a proteomic snapshot of the body. Various mathematical models are applied to large-scale clinical data to establish algorithms with predictive value for cardiovascular health, metabolic rate, lean body mass, liver inflammation, cardiorespiratory fitness, and glucose tolerance.

[0337] Enhanced Liver Fibrosis (ELF)

[0338] The ELF assay is a non-invasive blood test that measures three markers of liver inflammation and fibrosis: hyaluronic acid, procollagen III amino-terminal peptide (PIIINP) and tissue inhibitor of matrix metalloproteinase 1 (TIMP-1). The levels of these three markers, when used in conjunction with accompanying clinical data, have been shown to be highly predictive of liver inflammation and fibrosis symptoms when correlated with histological data in larger clinical trials.

[0339] Exploratory biomarkers

[0340] Blood samples will be collected and separated into plasma and serum components, from which 400 μL aliquots will be taken, labeled, and stored for future analysis of proteins, lipids, or gene expression that may help explain the pharmacological action of compound 1.

[0341] Eye Examination

[0342] To determine the subject's baseline status and to monitor changes from baseline during the treatment period, a complete eye examination including fundus photography of the posterior pole of the eye, OCT of the macula of both eyes, and slit lamp examination will be performed at screening and at the end of treatment (approximately day 61). Pupil dilation will be performed with 2.5% neosynephrine and 0.5% tropicamide (Mydriacyl), one drop per eye (unless contraindicated by the ophthalmologist), once for light-colored eyes and up to two times at 5-minute intervals for dark-colored eyes. A slit lamp is a biomicroscope with a bright light used in eye examinations to evaluate the front of the eye and various structures within the eye to determine health and detect eye diseases. OCT is a non-invasive imaging technique that uses light waves to take cross-sectional pictures of the retina. Fundus photography takes pictures of the back of the eye. These tests are standard tests involved in the medical evaluation of eye health and should be performed in collaboration with a limited number of ophthalmologists to ensure consistency of evaluation.

[0343] Vital signs

[0344] Vital signs include temperature, systolic blood pressure, diastolic blood pressure, heart rate, and respiratory rate. All blood pressure measurements must be taken with a blood pressure cuff that fits the subject's arm. A blood pressure cuff that is too small will result in inaccurately high blood pressure readings. Blood pressure and heart rate are recorded after the subject has been in the supine position for at least five minutes. Three blood pressure measurements are taken at each time point, with each measurement approximately two minutes apart. The first blood pressure is discarded. The second and third blood pressure measurements are entered into a database and averaged to produce the value for that time point.

[0345] Weight, muscle and body fat will be measured using an InBody scale and should be done pre-dose, in a fasting state and at approximately the same time of day.

[0346] Body temperature will also be monitored daily at home using a Braun Thermoscan 7 cochlear thermometer provided to the subject. This thermometer displays temperature in a color-coded format, indicating normal, elevated temperature (greater than 99.9°F, indicated as yellow) or fever (greater than 103°F, indicated as red). It has an audible feedback system to ensure proper use and alert the user that a temperature has been taken. The last nine readings are recorded on the thermometer. Temperatures should be taken daily and at the time of dosing. If subjects have symptoms suggestive of fever, they should have their temperature taken and confirmed. If a yellow or red indication is present and the temperature is elevated ≥100°F, subjects should discontinue taking Compound 1, avoid antipyretics (acetaminophen, aspirin or nonsteroidal anti-inflammatory drugs) and contact the investigational site.

[0347] Lab Test

[0348] Screening only: Viral serological tests include testing for the presence of hepatitis B antigen, anti-hepatitis C antibody, and anti-HIV antibody. TSH and free T4. eGFR=CKD-EPI creatinine equation (NKF 2009; www.kidney.org / content / ckd-epi-creatinine-equation-2009).

[0349] Hematology tests included mean corpuscular hemoglobin concentration (MCHC), mean corpuscular volume (MCV), hematocrit, hemoglobin, white blood cell count, and absolute lymphocyte, monocyte, neutrophil, basophil, eosinophil and platelet counts.

[0350] Serum chemistry analyses included glucose, calcium, albumin, total protein, sodium, potassium, bicarbonate, chloride, magnesium, blood urea nitrogen (BUN), creatinine, alkaline phosphatase, phosphate, uric acid, lactate dehydrogenase, ALT, AST, gamma-glutamyltransferase (GGT), bilirubin (total and direct bilirubin), amylase, and CPK. Baseline ALT and AST values ​​will be recorded, but for statistical evaluation of these parameters in exploratory analyses, the mean of the screening value and the pre-dose value on Day 1 will be used as the baseline value.

[0351] The lipid panel includes total cholesterol, HDL, LDL, VLDL, triglycerides and FFAs.

[0352] Additional tests at specific time points include glycated albumin, hs-CRP, ApoB, Lp(a) and HOMA-IR (which includes glucose, insulin, C-peptide; Wallace 2004), as well as PEth testing. For HOMA-IR, blood samples will be taken three times for each of the three analyses (blood glucose, serum insulin, and C-peptide) with a minimum of 5 minutes between each sample. The PEth test is a serum biomarker that can assess recent alcohol intake. It depends on both the amount of alcohol consumed and the time since consumption. The test will be evaluated at screening and on day 28. It may be performed at other times, at the discretion of the investigator, if there is concern about excessive alcohol intake based on medical history, symptoms, or laboratory evaluation (e.g., elevated liver function tests).

[0353] Urinalysis is performed by dipstick testing, followed by speckle microscopy if the dipstick shows little (1+), moderate (2+), or large (3+) amounts of blood or protein. If two urine collections show more than trace amounts of protein, spot tests for urine protein and albumin are performed.

[0354] Female subjects will be administered a urine pregnancy test.

[0355] Laboratory tests may be repeated once at Screening. Additional laboratory evaluations may be performed at the discretion of the investigator if medically justified upon evaluation of an adverse event.

[0356] 12-lead electrocardiogram

[0357] A single 12-lead ECG measurement will be taken after the subject has rested in the supine position for at least 10 minutes. External stimuli should be kept to a minimum. Video games, television viewing, and conversation are prohibited during this time. A digital ECG device will be used for the test. If the ECG time overlaps with any blood samples, the blood samples will be taken at the scheduled time points and the ECG will be taken ±10 minutes from that time point. Additionally, whenever possible, subjects should not eat within 2 hours prior to the ECG being performed.

[0358] ECG will be measured using an ECG device that automatically calculates heart rate and measures PR, RR, QRS, QT, and QTcF (Fridericia correction formula). Whenever possible, the same ECG device should be used for the same subjects throughout the study. ECG examinations must be performed in accordance with the sponsor-approved study unit SOPs.

[0359] Example 6: Exploratory Phase 2A Study of Compound 1 for the Treatment of Subjects with Obesity Heart Failure with Preserved Ejection Fraction (HFpEF)

[0360] The study is a Phase 2A randomized, parallel-group, placebo-controlled, double-blind, within-subject, dose-escalation study of compound 1 and placebo at three dose levels. An estimated 62 participants will be enrolled. Subjects will be randomized (1:1) to receive either compound 1 or placebo. Two dose levels (150 mg / day, then 300 mg / day) will be administered sequentially for 20 days each, reaching a third and highest dose of 450 mg / day if safety and tolerability are demonstrated at the previous two doses. The 450 mg high dose will continue for a total of 94 days, with a safety follow-up visit occurring within 14 days of the final dose.

[0361] Subjects will be screened for 40 days to determine eligibility based on specific medical history, physical examination, laboratory tests, and imaging evaluations according to an evaluation schedule. A single clinical site visit for screening is planned, but scheduling issues may necessitate additional visits to complete screening procedures. Many of these evaluations will be baseline before drug administration. A central laboratory will be used for all evaluations, including MRI, DEXA, clinical blood / plasma measurements, transthoracic echocardiography, and CPET.

[0362] Compound 1 is being evaluated for its effectiveness in improving cardiovascular function in obese subjects with heart failure with preserved ejection fraction (HFpEF).

[0363] Inclusion criteria: 1. Adult male or female aged 40 or older. 2. Be capable of understanding the information contained in an Institutional Review Board (IRB) or Independent Ethics Committee (IEC) approved Informed Consent Form (ICF) and must sign the form prior to the start of any study procedures. 3.Body mass index (BMI)>30kg / m2. 4. Signs and symptoms of HF as determined by the investigator, meeting the following severity criteria: a. KCCQ OSS<80; b. NYHA classification class II-III; c. baseline peak VO2 <18 mL / kg / min in women and <20 mL / kg / min in men; d. baseline respiratory exchange ratio (RER[RQ]) >1.0; e. left ventricular ejection fraction (EF) >50%; f. at least one of the following objective criteria for HF: i. documented hospitalization for HF within the past year or, if past year or more, documented with additional structural heart disease on echocardiography (enlarged left atrial volume or left ventricular hypertrophy, with sex-specific cut points according to Lang, 2015): · Left ventricular hypertrophy (LVH): a. Male: Septal wall thickness (cm) > 1.1 or posterior wall thickness > 1.1; b. Female: Septal wall thickness (cm) >1.0 or posterior wall thickness >1.0; Left Atrial Dilation (LAD): AP dimension (cm): >4.0 in men; >3.8 in women; ii. Pulmonary capillary wedge pressure (PCWP) >15mmHg at rest (or left ventricular end-diastolic pressure [LVEDP] >18mmHg) or >25mmHg with exercise (or 2.0mmHg / L / min) in the past year; iii. E / e' ratio of septal annulus at rest >14 on Doppler and tissue Doppler images in the past year; or iv. Currently elevated NT-proBNP, defined as >125pg / mL in subjects without atrial fibrillation / flutter and >375pg / mL in subjects with atrial fibrillation / flutter. 5. Participants must maintain a stable level of physical activity for the duration of the study and agree not to participate in any exercise training programs during the study. 6. Participants must maintain a stable diet and have no plans to participate in a weight loss program before or during the study. 7. Euthyroid function as assessed by thyroid profile with thyroid stimulating hormone (TSH) and free thyroxine (T4) tests at screening. Subjects with a stable history of thyroid disease and taking stable doses of thyroid medication for at least 4 months are eligible to be enrolled. 8.Ambulatory (not dependent on wheelchair or scooter) and able to perform upright exercise testing including 6MWT. 9. Stable (defined as no new medications or ≥50% change in existing medications) medications in the 30 days prior to screening, with the following additional special criteria for diuretics: a. Patients receiving loop or thiazide diuretic therapy must be on a stable regimen with flexibility for rescheduling of diuretic dosing.

[0364] Exclusion criteria: 1. Life expectancy less than 1 year due to non-cardiovascular reasons as determined by the investigator. History of malignancy within 2.5 years (excluding non-high-grade skin cancer, carcinoma in situ, or low-grade prostate cancer). 3. Weight change (gain or loss) of 10 pounds or more within the past 90 days (self-reported or recorded). 4. Bariatric surgery before screening or scheduled bariatric surgery during the study period. 5.Initiation of treatment with a GLP-1 receptor antagonist within 1 year of screening. 6.Start of SGLT2 inhibitor treatment within 6 months of screening. 7. MRI intolerance or any condition that contraindicates MRI scanning, including but not limited to: a. Wearing surgical clips / metal implants / shrapnel / internal electrical implants; or b. The subject cannot fit into an MRI scanner due to his or her size or the weight capacity of the scanner (typically 350 or 400 pounds, depending on the manufacturer); or c. Claustrophobia: History of severe claustrophobia that would prevent an MRI from being performed. 8. Acute compensated HF requiring ongoing IV diuretics or recent (<1 month prior to screening) hospitalization for HF. 9. Primary cardiomyopathy (e.g., constrictive, restrictive, infiltrative, toxic, hypertrophic [congenital], congenital, or other primary cardiomyopathies as determined by the investigator). 10. Ongoing myocarditis (COVID-induced or otherwise). 11. Active collagen vascular disease. 12. Active moderate or greater left or right sided valvular disease in the opinion of the investigator. 13. Scheduled cardiac surgery or catheterization during the study participation period. 14. Record of EF < 40% within the past 3 years. 15. Tachycardia (>110 beats / min) at screening. 16. Atrial fibrillation or atrial flutter with uncontrolled heart rate response or resting heart rate >110 bpm on screening electrocardiogram. Subjects may be rescreened after appropriate medication adjustments to manage atrial fibrillation. Up to 16 subjects with this condition may be enrolled in this study. 17. Untreated, life-threatening arrhythmia. Phase 2a study results The Phase 2a metabolism study of compound 1 was a 61-day randomized, double-blind, placebo-controlled study in obese subjects with high liver fat (≥8%) (body mass index 28-45 kg / m 2 The Phase 2a study was designed to evaluate the safety and efficacy of three dose levels of Compound 1 (150 mg, 300 mg, and 450 mg) in a randomized controlled trial. Eighty participants, ranging in age from 28 to 65 years, were randomly assigned to one of three Compound 1 treatment arms or matching placebo, stratified and blocked by HbA1C levels ≥ 5.7%, and administered once daily (fasted). Participants were instructed not to change their dietary or exercise behaviors. The Phase 2a study achieved its primary endpoint (liver fat reduction by MRI-PDFF) and secondary endpoints (weight and fat reduction by abdominal MRI). Key results and discussion include the following: There was a statistically significant reduction in liver fat at all three dose levels (p<0.0001 by ANCOVA). The relative reduction in liver fat was 33%, 43%, and 40%, with responder rates (relative reduction of >30%) of 40%, 71%, and 72% for the low, medium, and high doses, respectively. The relative reduction in liver fat for placebo was 2%, and the responder rate was 5%. Key cardiovascular and metabolic health indices observed across all dose levels were: Glycosylated albumin, an indicator of glucose control and insulin function, was reduced in a dose-dependent manner (p<0.0001, high dose). ○ There was a dose-dependent decrease in the inflammatory marker high-sensitivity C-reactive protein (hsCRP), an important parameter for cardiovascular risk (p<0.005, high dose). Compound 1 was well tolerated at all dose levels, with good compliance. No serious adverse events or deaths were reported. Diarrhea and transient flushing associated with alcohol intake were observed in 25% and 31.6% of Compound 1 subjects, respectively, and were the most commonly reported emergent adverse events. Most of these adverse events were mild, and one participant in the low-dose group discontinued Compound 1 due to diarrhea, whereas no participants in the high-dose group discontinued for any reason. Specific efficacy and safety results from Phase 2a are shown in the following figures and tables: (1) As shown in Table 5, Compound 1 was well tolerated over an 8-week period. [Table 5] (2) Body temperature changes as shown in Figure 13 and Table 6. [Table 6] (3) PK results (mean ± SEM) as shown in Figure 14 . (4) As shown in Figure 15 and Table 7, therapeutic effects were observed at all doses. Placebo-corrected percent change from baseline in MRI-proton density fat fraction (PDFF). [Table 7] (5) As shown in Figure 16 and Table 8, therapeutic effects were observed at all doses. Analysis of covariance of MRI-proton density fat fraction (PDFF). Mean change from baseline (LS mean ± 95%) at day 61 in the FAS population [Table 8] (6) As shown in Table 9, a therapeutic effect (a reduction in liver fat of more than 30% by MRI-PDFF) was observed in all treatment groups. [Table 9] (7) Significant weight loss at doses of 300 mg and 450 mg of Compound 1 administered once daily, as shown in FIG. 17 and Table 10. Repeated measures analysis of InBody weight. Mean change from baseline FAS population (LS mean ± 95% CI). [Table 10] (8) Observed body weight as shown in Figure 18 and Table 11. [Table 11] (9) Visceral adipose tissue—observational data of the FAS population as shown in FIG. 19 and Table 12. Placebo-corrected changes from baseline in abdominal MRI liver volume and fat fraction in the FAS population by treatment group (mean ± SEM). [Table 12] (10) Subcutaneous adipose tissue observation data of the FAS population, as shown in Figure 20 and Table 13. Placebo-corrected changes from baseline in abdominal MRI liver volume and fat fraction in the FAS population by treatment group (mean ± SEM). [Table 13] (11) Confirmation of fat loss (total adipose tissue) by MRI as shown in Figure 21. (12) Liver volume observation data for the FAS population as shown in Figure 22 and Table 14. [Table 14] (13) Systolic blood pressure observed as shown in FIG. 23. Mean change from baseline at day 61 in the FAS population (mean ± SEM) (14) Diastolic blood pressure observed as shown in Figure 24. Mean change from baseline at day 61 in the FAS population (mean ± SEM) (15) High sensitivity C-reactive protein (hsCRP) observed as shown in Figure 25 and Table 15. Mean change from baseline at day 61 for the FAS population (LS mean ± 95CI). [Table 15]

Claims

1. A composition for use in a method of reducing cardiovascular risk or mortality in a subject suffering from symptoms due to cardiovascular disease, said composition comprising 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof, said method comprising administering to said subject a therapeutically effective amount of 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof.

2. A) said symptoms are shortness of breath, shortness of breath during exertion, dizziness, chest pain, fainting, fatigue, cardiac energy impairment, or limitation of activities of daily living; optionally said limitation of activities of daily living is difficulty with self-care, mobility, or eating; B) said cardiovascular disease includes heart failure, myocardial infarction, coronary artery disease, or coronary heart disease (CHD); optionally said heart failure includes heart failure with preserved ejection fraction (HFpEF), heart failure with reduced ejection fraction (HFrEF), or heart failure with mildly reduced ejection fraction (HFmrEF); or C) said subject experiences a reduction in the risk of serious cardiovascular events after administration; optionally said serious cardiovascular event is death or hospitalization due to disease exacerbation, The composition according to claim 1.

3. A composition for use in a method of treating HFpEF, HFrEF or HFmrEF in a subject, said composition comprising 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof, said method comprising administering to said subject a therapeutically effective amount of 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof.

4. Said subject is A) suffering from at least one of the symptoms selected from shortness of breath, shortness of breath during exertion, cardiac energy impairment, dizziness, fatigue, dyspnea, palpitations (atrial fibrillation), chest discomfort, edema, fainting, and limitation of activities of daily living; optionally said limitation of activities of daily living is difficulty with self-care, mobility and eating; B) suffering from at least one symptom selected from reduced exercise tolerance, fatigue, lethargy, increased recovery time after exercise, and ankle swelling; or suffering from at least one symptom selected from coronary artery disease, hypertension, and heart murmur The composition according to claim 3.

5. A composition for use in a method of reducing blood pressure in a subject, said composition comprising 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof, said method comprising administering to said subject a therapeutically effective amount of 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof; Optionally, reducing said blood pressure comprises reducing diastolic blood pressure and / or reducing systolic blood pressure; Optionally, said method reduces or delays the progression of the risk of developing cardiovascular disease or heart failure. Composition.

6. Said subject is A) suffering from cardiovascular disease, hypertension, resistant hypertension, or severe hypertension, Optionally, said cardiovascular disease includes heart failure, heart attack, coronary artery disease, or coronary heart disease (CHD), Optionally, heart failure includes HFpEF, HFrEF, or HFmrEF; B) having hypertension associated with HFpEF, HFrEF, or HFmrEF; C) suffering from at least one symptom selected from headache, shortness of breath, chest pain, nosebleed, dizziness, fatigue, visual impairment, arrhythmia, hematuria, sweating, sleep disorder, and eye blood spots; D) experiencing a blood pressure reduction of at least 5 mmHg after administration; E) suffering from at least one symptom selected from headache, shortness of breath, chest pain, nosebleed, dizziness, fatigue, visual impairment, arrhythmia, hematuria, sweating, sleep disorder, and eye blood spots The composition according to claim 5.

7. A composition for use in a method of treating cardiovascular disease in a subject, said composition comprising 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof, said method comprising administering to said subject a therapeutically effective amount of 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof; said method achieving i) A steady state of the maximum plasma concentration (C ｍａｘ ) of 2,4-dinitrophenol from about 80 ng / mL to about 8300 ng / mL; ii) an average half-life (t １／２ ) of 2,4-dinitrophenol of from about 20 to 50 hours, from about 25 to 40 hours, or from about 30 to 40 hours; １／２ ) iii) The median time to reach the maximum plasma concentration (T ｍａｘ ) of 2,4-dinitrophenol of about 6 to 8 hours or about 6 to 10 hours; iv) The median value of the area under the curve extrapolated to infinity (AUC) of 2,4-dinitrophenol from about 3 h*μg / mL to about 420 h*μg / mL ｉｎｆ ); and v) AUC / C of about 18 ｍａｘ Ratio at least one of **Claim 8**: The composition according to any one of claims 1, 5 or 7, wherein the subject has obesity, excess body fat, diabetes, hypertension (hypertensive disease), dyslipidemia, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, or metabolic syndrome. **Claim 9**: i) A method for treating mitochondrial-related disorders or symptoms in a subject without causing a clinically significant risk of adverse events; or ii) A composition for use in a method for reducing toxicity or side effects or preventing overdose in treating mitochondrial-related disorders or symptoms in a subject, the composition comprising 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof, the method comprising administering to the subject a therapeutically effective amount of 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof; optionally, the overdose is associated with a mitochondrial uncoupling agent; optionally, the mitochondrial uncoupling agent is 2,4-dinitrophenol, a composition. **Claim 10**: A) The disorder is obesity, excess body fat, diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, cardiovascular disease, atherosclerosis, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, Rett syndrome, metabolic syndrome associated with aging, metabolic diseases associated with increased reactive oxygen species (ROS), Friedreich's ataxia or liver disease; optionally, the diabetes is type 2 diabetes (T2DM); B) The cardiovascular disease includes heart failure, HFpEF, HFrEF, HFmrEF, heart attack, coronary artery disease or CHD; C) The liver disease includes non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), non-cirrhotic NASH, non-cirrhotic NASH with hepatic fibrosis, hepatic steatosis, hepatic fibrosis, cirrhosis or hepatocellular carcinoma; D) The symptoms are at least one of steatosis, inflammation, fibrosis, cirrhosis and hepatocyte injury in NASH, the composition according to claim 9. **Claim 11**: The method is i) Extend the half-life (t １／２ ) of 2,4-dinitrophenol; ii) Delay the time (T ｍａｘ ) until the plasma concentration of 2,4-dinitrophenol reaches its maximum; iii) reducing the maximum plasma concentration (C ｍａｘ ) of 2,4-dinitrophenol; and, iv) increasing the area under the curve (AUC); or wherein the method, after administration, in the subject, v) a steady state of the maximum plasma concentration (Cmax) of 2,4-dinitrophenol from about 80 ng / mL to about 8300 ng / mL; vi) an average half-life (t1 / 2) of 2,4-dinitrophenol of about 20 - 50 hours, about 25 - 40 hours, or about 30 - 40 hours; vii) a median time for 2,4-dinitrophenol to reach the maximum plasma concentration (Tmax) of about 6 - 8 hours or about 6 - 10 hours; viii) a median value of the area under the curve extrapolated to infinity (AUCinf) of 2,4-dinitrophenol from about 3 h*μg / mL to about 420 h*μg / mL; and, ix) an AUC / Cmax ratio of about 18 and provides at least one of the composition according to claim 9 or claim 10. **Claim 12** A composition comprising 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof, A) the composition is for use in a method for increasing the metabolic rate without causing a clinically significant risk of adverse events in a subject, the method comprising administering to the subject a therapeutically effective amount of 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof; optionally the resting metabolic rate increases by at least 10% or at least 20%; or B) the composition is for use in a method for increasing the resting energy consumption of a subject, the method comprising administering to the subject a therapeutically effective amount of 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof; optionally the subject experiences an increase in resting energy consumption of at least 10%, at least 20% or about 30% after administration, composition. A composition for use in a method for treating metabolic disorders, said composition comprising 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof, said method comprising administering a therapeutically effective amount of 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof, to a subject. A composition comprising 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof, a) said composition is for use in a method for treating hypertriglyceridemia associated with cardiovascular disease, atherosclerosis, obesity, hypertension, diabetes, insulin resistance, and / or liver disease in a subject, said method comprising administering a therapeutically effective amount of 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof, to said subject; or b) said composition is for use in a method for treating severe hypertriglyceridemia in a subject, said method comprising administering a therapeutically effective amount of 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof, to said subject; optionally, said subject suffers from at least one of abdominal pain, pain in the mid-abdominal, chest, or back regions, gastrointestinal pain, dyspnea, anorexia, nausea, vomiting, inflammation of the pancreas, memory loss, dementia, xanthoma striatum, corneal arcus, and xanthoma; Composition. **Claim 15**: The composition according to any one of claims 12 to 14, wherein the subject suffers from at least one of obesity, excessive body fat, type 2 diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, atherosclerotic disease, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, Rett syndrome, metabolic syndrome associated with aging, metabolic diseases associated with an increase in reactive oxygen species (ROS), Friedreich's ataxia, NAFLD, NASH, non-sclerotic NASH, non-sclerotic NASH with hepatic fibrosis, hepatic steatosis, hepatic fibrosis, liver cirrhosis, and hepatocellular carcinoma. **Claim 16**: The method does not cause a clinically significant risk of adverse events in the subject after administration; Optionally; A) The adverse event includes at least one of nausea, vomiting, sweating, dizziness, headache, cataract, glaucoma, fever, hyperthermia, tachycardia, sweating, tachypnea, and death; B) The adverse event is characterized by at least one of increased body temperature, increased heart rate, abnormal sweating, erythema, sweating, dehydration, and abnormally rapid breathing; C) The adverse event is associated with cardiovascular collapse, cardiac arrest, and / or death; or D) The adverse event is associated with cardiac arrest; Optionally, the adverse event is associated with a mitochondrial uncoupling agent; optionally, the mitochondrial uncoupling agent is 2,4-dinitrophenol. The composition according to any one of claims 1 to 7, 9, 10, and 12 to 14. **Claim 17** A composition for use in a method for treating obesity, excess body fat, type 2 diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, arteriosclerosis, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, Rett syndrome, metabolic syndrome associated with aging, metabolic diseases associated with an increase in reactive oxygen species (ROS), Friedreich's ataxia, NAFLD, NASH, non-cirrhotic NASH, non-cirrhotic NASH with hepatic fibrosis, hepatic steatosis, hepatic fibrosis, cirrhosis, cancer, or hepatocellular carcinoma, wherein the composition comprises 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof, and the method comprises administering a therapeutically effective amount of 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof, to a subject; wherein the method i) A steady state of the maximum plasma concentration (C ｍａｘ ) of 2,4-dinitrophenol from about 80 ng / mL to about 8300 ng / mL; ii) An average half-life (t １／２ ) of 2,4-dinitrophenol of about 20 to 50 hours, about 25 to 40 hours, or about 30 to 40 hours; iii) The median time for 2,4-dinitrophenol to reach the maximum plasma concentration (T ｍａｘ ) of about 6 to 8 hours or about 6 to 10 hours; iv) The median value of the area under the curve extrapolated to infinity (AUC) of 2,4-dinitrophenol from about 3 h*μg / mL to about 420 h*μg / mL ｉｎｆ ); and v) AUC / C of about 18 ｍａｘ Ratio achieves at least one of the following, composition. **Claim 18** wherein the subject A) has a heightened body mass index (BMI); optionally the subject has a BMI of from about 28.0 kg / m2 to about 45.0 kg / m2; and / or B.1) is in a fasting state prior to administration; or B.2) is in a fed state prior to administration, The composition according to any one of claims 1-7, 9, 10, 12-14 and 17. **Claim 19** wherein the therapeutically effective amount is A) from about 30 mg to about 1400 mg per day, from about 100 mg to about 1000 mg per day, from about 150 mg to about 600 mg per day, or from 200 mg to 550 mg per day; B) about 30 mg, 50 mg, 75 mg, 100 mg, 150 mg, 170 mg, 200 mg, 250 mg, 300 mg, 340 mg, 350 mg, 400 mg, 450 mg, 500 mg, 510 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1050 mg, 1100 mg, 1150 mg, 1200 mg, 1250 mg, 1300 mg, 1350 mg or 1400 mg per day; C) about 30 mg, 100 mg, 200 mg, 500 mg, 600 mg, 1050 mg or 1400 mg per day; D) about 200 mg, 400 mg or 550 mg per day; E) Approximately 170 mg, 340 mg, 510 mg per day; or F) Approximately 150 mg, 300 mg, 450 mg per day The composition according to any one of claims 1 to 7, 9, 10, 12 to 14 and 17. **Claim 20** The 5-[(2,4-dinitrophenoxy)methyl]-1-methyl-2-nitro-1H-imidazole, or a pharmaceutically acceptable salt thereof, is a) Administered orally once a day; and / or b) Administered once a day for 2 weeks, 4 weeks, 6 weeks, 8 weeks, 10 weeks, 1 month, 2 months, 3 months, 4 months, 6 months, 8 months or 1 year The composition according to any one of claims 1 to 7, 9, 10, 12 to 14 and 17, characterized by the above.

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