CN117881399A

CN117881399A - Methods of treating associated conditions of the granulear body

Info

Publication number: CN117881399A
Application number: CN202280042269.9A
Authority: CN
Inventors: S·坎; D·约卡斯基
Original assignee: Levis Pharmaceutical Co ltd
Current assignee: Levis Pharmaceutical Co ltd
Priority date: 2021-05-20
Filing date: 2022-05-19
Publication date: 2024-04-12

Abstract

Disclosed herein are methods of treatment comprising administering to a subject in need thereof a therapeutically effective amount of 5- [ (2, 4-dinitrophenoxy) methyl ] -1-methyl-2-nitro-1H-imidazole or a pharmaceutically acceptable salt thereof. 5- [ (2, 4-dinitrophenoxy) methyl ] -1-methyl-2-nitro-1H-imidazole is useful in the treatment of granline-related disorders or conditions, including obesity, diabetes, hypertension, cardiovascular diseases, and liver diseases.

Description

Methods of treating associated conditions of the granulear body

Technical Field

The present disclosure relates to methods for treating cardiovascular diseases and granulear-associated disorders or conditions without causing adverse events or clinically significant risks of overdosing.

Background

Cardiovascular diseases are the types of diseases involving the heart and blood vessels, such as coronary artery disease, heart attacks, strokes, heart failure, hypertensive heart disease, and rheumatic heart disease. In the united states, over 650 ten thousand people suffer from heart failure (Cardiology Today, 4 months, 6 days in 2017). Heart failure with preserved ejection fraction (HFpEF), also known as diastolic heart failure, results in almost half of the us 650-thousand heart failure cases. HFpEF is caused by abnormalities in active ventricular relaxation and passive ventricular compliance, resulting in a decline in stroke volume and cardiac output (Am Fam Physician.2017Nov 1;96 (9): 582-588). In heart failure with reduced ejection fraction (HFrEF), also known as systolic heart failure, the myocardium is unable to contract sufficiently and less oxygen-enriched blood is expelled into the body. Mortality was similar between patients with HFpEF and HFrEF (Cardiology Today,2017, 4, 6). Heart failure with moderate ejection fraction (HFmrEF) is a new category of heart failure between HFpEF and hfrlef. HFmrEF has a incidence of 10-20% of heart failure patients (Maedica (Bucur), 2016,11 (4): 320-324). Thus, there is a great need for an effective treatment of heart failure including HFpEF, hfaref and HFmrEF.

The granulosomes control metabolism in individual cells by burning sugar and fat. Granulosa decoupling is a robust and natural process for the body to generate heat. Granulosomes are uncoupled from ATP phosphorylation (complex V) by respiration (complexes I-IV) to generate heat. In fact, 20-40% of the calories consumed are used to produce body temperature. When the granulosa body fails to produce enough energy for the body to function properly, a granulosa body-related disorder or condition occurs, affecting almost any part of the body, including cells of the brain, adipose tissue, nerves, muscles, heart, lung, liver, kidneys, pancreas, eyes, and ears.

For many years, the administration of granulin chemical uncoupling agents as a means of reducing fat deposition has been a scientific goal. Although there are a variety of small molecules that uncouple the oxidative phosphorylation of granosomes, the most well known small molecule is 2, 4-Dinitrophenol (DNP). Although DNP is known to have robust uncoupling, it is unfortunately associated with unacceptably high proportions of significant adverse effects (J.Med. Toxicol.2011, month 9; 7 (3): 205-212). These adverse effects may include hyperthermia, tachycardia, sweating, and tachypnea, ultimately leading to death. As a small highly permeable lipophilic acid, DNP is rapidly absorbed in the stomach. The high concentration rapidly distributes and uncouples immediately, generating a large amount of heat in a short time. Therefore, DNP has a small therapeutic index and is extremely dangerous when overdosed. DNP is labeled by the 1938 federal food, drug, and cosmetic laws as "extremely dangerous and unsuitable for human consumption". Thus, there is a need for decoupling agents that can safely treat a mitochondrial related disorder or condition.

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 Accelerator (CMA). It is designed to effectively address the root cause of metabolic diseases, i.e., the accumulation of fat and sugar in the body. CMA helps improve cellular metabolism and increase energy consumption and calorie consumption, reducing fat accumulation. By using a new controlled and targeted approach, compound 1 can increase granulomatous body leakage, an in vivo persistence process that consumes energy and accounts for 20% -40% of calories per day. Compound 1 utilizes a granulear uncoupling mechanism to increase substrate utilization.

Compound 1 has been studied using a non-clinical model. Demonstration of potent therapeutic activity in related rodent disease models, as well as pharmacokinetic and safety profiles, support compound 1 may be advantageously and safely used in the treatment of a wide range of granulomatous related diseases. Compound 1 has also been found to be effective in the treatment of cardiovascular disease.

Disclosure of Invention

In some embodiments, the present disclosure provides methods for treating cardiovascular disease in a subject comprising administering to the subject a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods of treating heart failure comprising administering to a subject a therapeutically effective amount of compound 1, or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods of reducing cardiovascular risk or mortality in a subject suffering from a symptom due to cardiovascular disease comprising administering to the subject a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof.

In some embodiments, the cardiovascular disease is selected from the group consisting of: heart failure, heart attacks, coronary artery disease, and Coronary Heart Disease (CHD).

Heart failure as used herein includes heart failure with a preserved ejection fraction (HFpEF) or heart failure with a reduced ejection fraction (hfrref) or heart failure with a moderate ejection fraction (HFmrEF).

In some embodiments, the cardiovascular disease is HFpEF.

In some embodiments, the cardiovascular disease is HFrEF.

In some embodiments, the cardiovascular disease is HFmrEF.

In some embodiments, the present 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 5- [ (2, 4-dinitrophenoxy) methyl ] -1-methyl-2-nitro-1H-imidazole or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods for treating heart failure with reduced ejection fraction (HFrEF) in a subject 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.

In some embodiments, the present disclosure provides a method for treating heart failure with moderate ejection fraction (HFmrEF) in a subject 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.

In some embodiments, the subject has at least one of shortness of breath, shortness of breath on exercise, impaired cardiac energy, dizziness, fatigue, dyspnea, palpitations (atrial tremor), chest discomfort, edema, syncope, and restricted activities of daily living.

In some embodiments, activities of daily living are limited by difficulties in personal care, mobility, and diet.

In some embodiments, the subject has a symptom selected from the group consisting of reduced exercise tolerance, fatigue, tiredness, increased recovery time after exercise, and ankle swelling.

In some embodiments, the subject has at least one of coronary artery disease, hypertension, and heart murmur.

In some embodiments, the present disclosure provides methods of reducing blood pressure in a subject comprising administering to the subject a therapeutically effective amount of compound 1, or a pharmaceutically acceptable salt thereof.

In some embodiments, the subject has at least one of cardiovascular disease, hypertension, refractory hypertension, and severe hypertension.

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

In some embodiments, the subject has high blood pressure associated with HFpEF.

In some embodiments, the subject has high blood pressure associated with HFrEF.

In some embodiments, the subject has high blood pressure associated with HFmrEF.

In some embodiments, wherein the subject experiences a decrease in blood pressure of at least 5mmHg after administration.

In some embodiments, the method reduces the risk of developing cardiovascular disease, and/or reduces the risk of HFpEF, HFrEF or HFmrEF.

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

In some embodiments, the method comprises at least one of:

i) Extending half-life of DNP (t) _1/2 )；

ii) delay of time to reach maximum plasma concentration of DNP (T) _max )；

iii) Reduction of maximum plasma concentration of DNP (C _max ) The method comprises the steps of carrying out a first treatment on the surface of the And

iv) increasing the area under the curve (AUC).

In some embodiments, the subject does not experience significant systemic toxicity, side effects, a significant increase in body temperature, or a significant increase in heart rate after administration.

In some embodiments, the present disclosure provides methods of treating cardiovascular disease by achieving the following goals:

i) A maximum plasma concentration (C) of DNP of about 80ng/mL to about 8300ng/mL _max ) Is a steady state of (2);

ii) an average half-life (t) of DNP of about 20-50 hours, about 25-40 hours or about 30-40 hours _1/2 )；

iii) Median time to maximum plasma concentration of DNP (T) of about 6-8 hours or about 6-10 hours _max )；

iv) extrapolated to infinite median Area Under Curve (AUC) of DNP from about 3h [ mu ] g/mL to about 420h [ mu ] g/mL _inf ) The method comprises the steps of carrying out a first treatment on the surface of the And

AUC/C of about 18 _max The ratio of the total number of the,

by administering to a subject from about 30mg to about 1400mg of compound 1 or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods of treating a subject's granulear-associated disorder or condition without causing clinically significant risk of an adverse event, the method comprising administering to the subject a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods of reducing toxicity or side effects in treating a subject's granline-related disorder or condition comprising administering to the subject a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods of preventing overdose in treating a subject's granulear-associated disorder or condition comprising administering to the subject a therapeutically effective amount of compound 1, or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods for increasing metabolic rate or resting energy expenditure in a subject without causing clinically significant risk of adverse events, comprising administering to the subject a therapeutically effective amount of compound 1, or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods for treating metabolic abnormalities in a subject comprising administering to the subject a therapeutically effective amount of compound 1, or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods of treating hypertriglyceridemia associated with cardiovascular disease, atherosclerosis, obesity, hypertension, diabetes, insulin resistance, and/or liver disease in a subject comprising administering to the subject a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods of treating severe hypertriglyceridemia in a subject comprising administering to the subject a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides a method of reducing liver fat in a subject by at least 50% or reducing lipid in a subject by at least 10%, the method comprising administering to the subject a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides a method of treating obesity, hyperlipidemia, 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-liver cirrhosis NASH with liver fibrosis, liver steatosis, liver fibrosis, liver cirrhosis, or hepatocellular carcinoma, the method comprising administering to a subject a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof so as to achieve at least one of:

i) A maximum plasma concentration (C) of 2, 4-dinitrophenol of about 80ng/mL to about 8300ng/mL _max ) Is a steady state of (2);

ii) an average 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 )；

iii) Median time to maximum plasma concentration (T) of 2, 4-dinitrophenol for about 6-8 hours or about 6-10 hours _max )；

iv) extrapolated area under the median curve (AUC) of about 3h [ mu ] g/mL to about 420h [ mu ] g/mL of 2, 4-dinitrophenol to infinity _inf ) The method comprises the steps of carrying out a first treatment on the surface of the And

v) AUC/C of about 18 _max Ratio.

Drawings

Figure 1 shows the plasma concentrations of 2, 4-dinitrophenol after administration of compound 1 and 2, 4-dinitrophenol in dogs.

Figures 2A and 2B show AUC of 2, 4-dinitrophenol after administration of compound 1.

Figures 3A and 3B show plasma compound 1 concentrations (linear and semilogarithmic scale) following oral administration of 500mg of compound 1 in food state.

Figures 4A and 4B show the plasma 2, 4-dinitrophenol concentration (linear and semilogarithmic scale) in food state after oral administration of 500mg of compound 1.

Fig. 5 shows the 2, 4-dinitrophenol curves for all groups.

Figure 6 shows body temperature of MAD groups during MAD QD dosing period.

Figure 7 shows Resting Energy Expenditure (REE) of MAD groups during the MAD QD dosing period.

Figure 8 shows weight loss following administration of compound 1.

Figure 9 shows the decrease in average glucose levels after administration of compound 1.

Figure 10 shows the change in systolic blood pressure after administration of compound 1.

Figure 11 shows the change in diastolic blood pressure after administration of compound 1.

Fig. 12 shows the heart rate observed after administration of compound 1.

Fig. 13 shows the observed change in body temperature after administration of compound 1.

Fig. 14 shows PK results observed after administration of compound 1.

Figure 15 shows the percentage of placebo corrected change in MRI Proton Density Fat Fraction (PDFF) from baseline value after administration of compound 1.

Fig. 16 shows an analysis of mean change in covariance of MRI Proton Density Fat Fraction (PDFF) from baseline on day 61 of FAS population.

Figure 17 shows the mean change from baseline for repeated measurement analysis of FAS population InBody body weight.

Figure 18 shows the observed change in body weight after administration of compound 1.

Figure 19 shows the percentage of placebo corrected change in abdominal MRI liver volume and obesity from baseline values for the FAS population of the treatment group.

Figure 20 shows the percentage of placebo corrected change in abdominal MRI liver volume and obesity from baseline values for the FAS population of the treatment group.

Fig. 21 shows fat loss (total adipose tissue) confirmed by MRI.

Fig. 22 shows the observed change in liver volume.

Figure 23 shows the average change from baseline in systolic blood pressure observed on day 61.

Figure 24 shows the average change from baseline in diastolic blood pressure observed on day 61.

Figure 25 shows the average change from baseline in hypersensitive C response protein (hsCRP) observed on day 61.

Detailed Description

Definition of the definition

While various embodiments and aspects of the present invention are shown and described herein, it will be apparent to those skilled in the art that these embodiments and aspects are provided by way of example only. Numerous variations, changes, and substitutions will now 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 employed in practicing the invention.

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 document parts cited in the application, including but not limited to patents, patent applications, articles, books, manuals, and treatises, are expressly incorporated by reference for any purpose.

5- [ (2, 4-dinitrophenoxy) methyl ] -1-methyl-2-nitro-1H-imidazole is a novel small molecule uncoupler. It has the following structure:

5- [ (2, 4-dinitrophenoxy) methyl ] -1-methyl-2-nitro-1H-imidazole can be prepared by the procedure described in WO 2018/129258.

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

Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. See, e.g., singleton et al, DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY, 2 nd edition, 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 in the practice of the present invention. The following definitions are provided to facilitate understanding of certain terms used frequently herein and are not intended to limit the scope of the present disclosure.

The term "a" or "an" as used herein means one or more.

The terms "comprising," "including," and "having," as well as derivatives thereof, are used interchangeably herein as a comprehensive, open-ended term. For example, the use of "including," "comprising," or "having" means that any element that includes, has, or comprises is not the only element encompassed by the subject of the clause containing the verb.

As used herein, the term "about" means a range of values that includes the specified value, which the skilled artisan considers reasonably similar to the specified value. In some embodiments, the term "about" means within standard deviation using measurements generally acceptable in the art. In some embodiments, "about" means a range extending to +/-10%, +/-5%, or +/-2% of the specified value. In some embodiments, "about" means the specified value.

As used herein, "treatment" or "alleviating" or "improving" or "reducing" are used interchangeably herein. These terms refer to methods of achieving a beneficial or desired result, including but not limited to therapeutic benefit. Therapeutic benefit means eradication or amelioration of the underlying disorder being treated. In addition, therapeutic benefit is achieved by eradicating or ameliorating one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the subject, although the subject may still have the underlying disorder. Treatment includes slowing the progression of clinical symptoms of the disease by administering the composition; throttling the disease, i.e., causing a decrease in clinical symptoms of the disease; inhibiting the disease, i.e., preventing the development of clinical symptoms by administering the composition after the initial appearance of symptoms; and/or alleviating the disease, i.e., causing regression of clinical symptoms by administering the composition after its initial appearance.

"patient" or "subject in need thereof" refers to a living organism that has or is susceptible to a disease or condition that can be treated by using the methods provided herein. The term does not necessarily indicate that a subject is diagnosed with a particular disease, but generally refers to an individual under medical supervision. Non-limiting examples include humans, other mammals, cattle, 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.

As used herein, "administering" a disclosed compound encompasses delivering a compound as described herein, or a prodrug or other pharmaceutically acceptable derivative thereof, to a subject using, for example, any suitable formulation or route of administration as described herein.

By "pharmaceutically acceptable" is meant compounds, salts, compositions, dosage forms, and other materials which are suitable for preparing a pharmaceutical composition suitable for veterinary or human medical use.

As used herein, the language "pharmaceutically acceptable salts" refers to salts of the applied 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 may be prepared from inorganic or organic acids. Examples of the inorganic acid include sulfuric acid, 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 the group of organic acids of the aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulphonic classes, 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, pamoic acid (pamoic acid), methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, pantothenic acid, trifluoromethanesulfonic acid, 2-hydroxyethanesulfonic acid, p-toluenesulfonic acid, sulfanilic acid, cyclohexylsulfamic acid, stearic acid, alginic acid, β -hydroxybutyric acid, salicylic acid, galactaric acid and galacturonic acid. Suitable pharmaceutically acceptable base addition salts of the compounds of the invention include, for example, ammonium salts and metal salts including alkali metal, alkaline earth metal and transition metal salts such as, for example, calcium, magnesium, potassium, sodium and zinc salts. Pharmaceutically acceptable base addition salts also include organic salts prepared from basic amines such as, for example, N' -dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and procaine. All of these salts can be prepared from the corresponding compounds by reacting, for example, an appropriate acid or base with the compound.

An "effective amount" is an amount sufficient to achieve the specified purpose (e.g., achieve the effect to which administration is intended, treat a disease, reduce enzymatic activity, reduce one or more symptoms of a disease or condition, reduce viral replication in a cell). An example of an "effective amount" is an amount sufficient to facilitate treatment, or reduce one or more symptoms of a disease, which may also be referred to as a "therapeutically effective amount". By "reducing" one or more symptoms (and grammatical equivalents of this phrase) is meant reducing the severity or frequency of the symptoms, or eliminating the symptoms. Efficacy may also be expressed as a "fold" increase or decrease. For example, a therapeutically effective amount may have at least 1.2-fold, 1.5-fold, 2-fold, 5-fold, or greater effect than a control.

As used herein, the term "significant increase in body temperature" in a subject refers to an increase in body temperature associated with deleterious effects on the subject, not limited to disease, 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 ℃, about 1 ℃, about 1.5 ℃, about 2 ℃, about 2.5 ℃, about 3 ℃, about 3.5 ℃, about 4.5 ℃, about 5 ℃, about 5.5 ℃, about 6 ℃ 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.

As used herein, the term "significant systemic toxicity" in a subject refers to systemic toxicity associated with deleterious effects on the subject, not limited to disease, physical discomfort or pain, coma, and death. In one non-limiting embodiment, significant systemic toxicity is indicated by an increase in the level of liver enzymes, blood urea nitrogen, or creatinine as compared to the corresponding level in the subject without the composition administered.

Therapeutic method

In some embodiments, the present disclosure provides a method of treating a cardiovascular disease in a subject comprising administering to the subject a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof. Preliminary studies have shown that compound 1 can achieve the following:

● Controlled and improved 2, 4-dinitrophenol PK-low C _max High AUC, low variability.

● Potentially administered orally once daily.

● Liver-specific uncoupling and reduced systemic effects.

● A broad therapeutic index.

Thus, compound 1 is useful for safely treating a wide range of diseases.

In some embodiments, the present disclosure provides a method of treating heart failure in a subject suffering from a symptom due to cardiovascular disease comprising administering to the subject a therapeutically effective amount of compound 1, or a pharmaceutically acceptable salt thereof.

In some embodiments, the symptom attributed to the cardiovascular disease is shortness of breath, dizziness, chest pain, syncope, fatigue, or restricted activities of daily living.

In some embodiments, activities of daily living are limited by difficulties in personal care, mobility, or diet.

In some embodiments, the present disclosure provides methods of treating cardiovascular disease in a subject comprising administering to the subject a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof.

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

Cardiovascular hemodynamic disturbances characterized by increased heart rate in patients lacking physical activity, increased risk of atrial fibrillation, increased blood volume, increased cardiac output, increased systemic vascular resistance, increased arterial pressure in sleep apnea patients, increased left ventricular wall pressure, increased pulmonary arterial pressure, and changes in ventricular pressure in patients with high blood pressure and insulin resistance.

Atherosclerosis and myocardial infarction, which may be increased indirectly by promoting significant risk factors for atherosclerosis (e.g., diabetes, hypertension, dyslipidemia) or directly by the steatosis endocrinopathy and immune diseases of epicardial adipose tissue.

Epicardial fat accumulation, pathogenic paracrine and vascular secretory signaling, inflammatory macrophage increase, T lymphocyte and obesity cell increase, adipogenic adipokine increase and vascular protective adipokine decrease

Heart Failure (HF), in particular HF with preserved ejection fraction (HFpEF)

Atherosclerotic cardiovascular disease (ASCVD), arrhythmia, heart fat infiltration, and coronary calcification.

Sleep apnea may cause hypoxia, elevated epinephrine (epinephrine) may cause hypertension, and intrathoracic pressure fluctuations increase left and right ventricular pressures.

Thrombotic and thromboembolic events, increased adipose tissue compresses the pelvis and lower limb veins, compromising venous return, and promoting deep vein thrombosis.

Abnormal cardiac cells and structures characterized by myocardial steatosis, apoptosis and fibrosis, as well as left ventricular remodeling and hypertrophy, left atrial enlargement, right ventricular hypertrophy, and increased adipose tissue surrounding the pericardium and blood vessels.

Reduced cardiac function characterized by hypoxia, atherosclerosis, thrombosis, left ventricular dysfunction (diastole and systole) and right ventricular failure due to sleep apnea.

Immune disorders characterized by an increase in pro-inflammatory adipocytes 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 adipocytes such as adiponectin and IL-10.

Immune disorders characterized by neutrophil activation and increased granulation tissue, such as severe asthma and severe asthma with glucocorticoid resistance.

Endocrinopathy characterized by activation of renin-angiotensin-aldosterone system leading to elevated blood pressure and altered expression of peroxisome proliferator-activated receptors

Endocrinopathy characterized by hyperinsulinemia, systemic insulin resistance and steatosis, and insensitivity to myocardial insulin.

Endocrinopathy, which is characterized by leptin insensitivity, elevated leptin levels can lead to cardiac hypertrophy and heart failure.

Lipotoxicity characterized by limited energy storage in surrounding subcutaneous adipose tissue.

Delivery extravasation of free fatty acids to liver, muscle, pancreas, kidney and/or visceral, pericardial and perivascular adipose tissue

In some embodiments, the cardiovascular disease is heart failure, heart attack, coronary artery disease, and Coronary Heart Disease (CHD).

In some embodiments, heart failure comprises HFpEF, hfaref or HFmrEF.

In some embodiments, the subject experiences a reduction in the risk of a significant cardiovascular event after administration.

In some embodiments, the major cardiovascular event is death or hospitalization for exacerbation of the disease.

In some embodiments, the present disclosure provides methods 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 pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods 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 pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods for treating heart failure with moderate ejection fraction (HFmrEF) in a subject comprising administering to the subject a therapeutically effective amount of compound 1, or a pharmaceutically acceptable salt thereof.

In some embodiments, the subject has obesity, hyperlipidemia, diabetes, hypertension, dyslipidemia, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, or metabolic syndrome.

In some embodiments, the subject has at least one symptom selected from the group consisting of: shortness of breath, shortness of breath on exercise, impaired cardiac energy, dizziness, fatigue, dyspnea, palpitations (atrial tremor), chest discomfort, oedema, syncope and restricted activities of daily living.

In some embodiments, the subject has at least one of the following: reduced exercise tolerance, fatigue, tiredness, increased recovery time after exercise, and ankle swelling.

In some embodiments, wherein the subject experiences an improvement in cardiac bioenergy deficiency after administration, wherein the improvement comprises a weight loss of >5%, a decrease in blood pressure, an increase in quality of life, an increase in exercise tolerance, and/or a decrease in risk of a major cardiovascular event, wherein the major cardiovascular event is selected from death, hospitalization for exacerbation of the disease, and myocardial infarction.

In some embodiments, the method further comprises assessing peak oxygen consumption (VO) of the subject during exercise before and after administering the therapeutically effective amount of compound 1 ₂ ) And/or VE/CO ₂ Or VE/VCO ₂ Slope, wherein VO in subject after administration ₂ An increase in HFpEF, HFrEF, HFmrEF in the subject is indicative of a decrease in the extent of HFpEF, HFrEF, HFmrEF, or a decrease in one or more symptomatic components or conditions of the cardiovascular disease thereof.

In some embodiments, the method increases VO in the subject after administration ₂ 。

In some embodiments, the method increases exercise tolerance in the subject.

In some embodiments, the method increases exercise tolerance in the subject as measured by assessing 6 minutes walking distance (6 MWD) before and after administration of a therapeutically effective amount of compound 1, wherein an increase in 6MWD in the subject after administration indicates a decrease in the level of HFpEF or at least one symptomatic component or condition thereof in the subject.

In some embodiments, the method increases the 6MWD after administration.

In some embodiments, HFpEF in a subject is diagnosed based on echocardiography (E/E') or a biomarker (NT-proBNP).

In some embodiments, the method further comprises evaluating the NYHA class score of the subject before and after administration.

NYHA functional classification rates the severity of heart failure symptoms as one of four functional categories. NYHA functional classification is widely used in clinical practice and research as it provides a standard description that can be used to assess the severity of response to treatment and guideline management. NYHA functions based on symptom severity and physical activity are classified as:

● Class I: physical activity is not limited. Normal physical activity does not cause excessive dyspnea, fatigue or palpitations

● Class II: physical activity is slightly restricted. While resting, it is comfortable, but normal physical activity can lead to excessive dyspnea, fatigue, or palpitations.

● Class III: physical activity is significantly restricted. Is comfortable at rest, but less than normal physical activity can lead to excessive dyspnea, fatigue, or palpitations.

● Class IV: no physical activity can be performed without discomfort. Symptoms can occur at rest. If any physical activity is performed, discomfort is increased.

In some embodiments, the method further comprises the steps of: the subject's NYHA classification score is assessed before and after administration of a therapeutically effective amount of compound 1, wherein a decrease in NYHA score after administration is indicative of a decrease in the extent of disease in the subject.

In some embodiments, the method reduces the NYHA class score of the subject after administration from class III to class II, or from class II to class I.

In some embodiments, the method increases the quality of life of the subject.

In some embodiments, the method improves the quality of life of a subject assessed by a standardized questionnaire, such as KCCQ (kansase cardiomyopathy questionnaire), KCCQ-12, KCCQ-body limit score (KCCQ-PLS), KCCQ-complete symptom score (KCCQ-TSS) KCCQ-clinical summary score (KCCQ-CSS), KCCQ-total summary score (KCCQ-OSS), or other derived questionnaires.

In some embodiments, the cardiovascular disease is selected from the group consisting of: heart failure, HFpEF, HFrEF, heart attacks, coronary artery disease, and Coronary Heart Disease (CHD).

In some embodiments, the subject has high blood pressure associated with HFpEF.

In some embodiments, the subject has high blood pressure associated with HFrEF.

In some embodiments, the subject has at least one symptom selected from the group consisting of: headache, shortness of breath, chest pain, nose bleeding, dizziness, fatigue, vision problems, arrhythmia, hematuria, sweating, sleep difficulties and eye bloodstains.

In some embodiments, the symptom is associated with HFpEF, hfaref or HFmrEF.

In some embodiments, wherein reducing blood pressure comprises reducing diastolic pressure and/or reducing systolic pressure.

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

In some embodiments, the method reduces the risk of developing cardiovascular disease, reduces the risk of HFrEF, or slows the progression of HFrEF.

In some embodiments, the method reduces the risk of developing cardiovascular disease, reduces the risk of HFmrEF, or slows the progression of HFmrEF.

In some embodiments, the subject is under fasting conditions prior to administration.

In some embodiments, the subject is in fed condition prior to administration.

In some embodiments, the subject experiences a decrease in at least one of body weight, blood pressure, and blood glucose after administration.

In some embodiments, the subject experiences at least one of:

i) Weight loss of at least 5% or at least 10%;

ii) a decrease in blood pressure of at least 5mmHg;

iii)HbA _1c a reduction of at least 0.5% or at least 1.5%;

iv) lipid reduction of at least 10%; and

v) liver fat reduction by at least 50%.

In some embodiments, the method comprises at least one of:

Extending half-life (t) of 2, 4-dinitrophenol _1/2 )；

Delay time to reach maximum plasma concentration of 2, 4-dinitrophenol (T _max )；

Reduction of the maximum plasma concentration (C) of 2, 4-dinitrophenol _max ) The method comprises the steps of carrying out a first treatment on the surface of the And

area under the curve (AUC) was increased.

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

In some embodiments, the median T of the 2, 4-dinitrophenol _max Extending to at least 6 hours or at least 8 hours.

In some embodiments, the median T of the 2, 4-dinitrophenol _max Extending to about 6-8 hours or about 6-10 hours.

In some embodiments, 2, 4-dinitrophenol C is reduced _max Comprising providing in the subject about 80ng/mL to about 8300ng/mL of C2, 4-dinitrophenol after administration _max Is a steady state of (c).

In some embodiments, the method provides an AUC/C of about 18 in the subject _max Ratio.

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

In some embodiments, the present disclosure provides methods of treating a cardiovascular disease comprising administering to a subject about 30mg to about 1400mg of compound 1, or a pharmaceutically acceptable salt thereof, so as to achieve at least one of:

v) AUC/C of about 18 _max Ratio.

In some embodiments, the present disclosure provides methods of reducing toxicity or side effects of 2, 4-dinitrophenol in treating a granline-related disorder or condition in a subject comprising administering to the subject a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods of preventing overdose of 2, 4-dinitrophenol in treating a subject's granulomatous related disorder or condition comprising administering to the subject a therapeutically effective amount of compound 1 or a pharmaceutically acceptable salt thereof.

In some embodiments, the granline-related disorder includes obesity, hyperlipidemia, diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, cardiovascular disease, atherosclerosis, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, retter's syndrome, metabolic syndrome associated with aging, metabolic disease associated with increased Reactive Oxygen Species (ROS), friedreich's ataxia, or liver disease.

In some embodiments, the disorder is a Branched Chain Amino Acid (BCAA) metabolic disorder, a lysosomal storage disorder, a glycogen storage disorder.

In some embodiments, the diabetes is type 2 diabetes (T2 DM).

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

In some embodiments, liver disease includes non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), non-cirrhosis NASH with liver fibrosis, liver steatosis, liver fibrosis, cirrhosis, or hepatocellular carcinoma.

In some embodiments, the granulomatous related disorder includes cardiovascular disease, hypertension, type 2 diabetes, dyslipidemia, obesity, or nonalcoholic steatohepatitis (NASH).

In some embodiments, the granulomatous related condition is at least one of steatosis, inflammation, fibrosis, cirrhosis, and hepatocyte damage in NASH.

In some embodiments, toxicity, adverse events, side effects, and overdosing are associated with the granosome decoupling agent.

In some embodiments, the granulometric decoupling agent is 2, 4-dinitrophenol.

In some embodiments, the method comprises at least one of:

i) Extending half-life (t) of 2, 4-dinitrophenol _1/2 )；

ii) delay of the time to reach the maximum plasma concentration of 2, 4-dinitrophenol (T) _max )；

iii) Reduction of the maximum plasma concentration (C) of 2, 4-dinitrophenol _max ) The method comprises the steps of carrying out a first treatment on the surface of the And

iv) increasing the area under the curve (AUC).

In some embodiments, the present disclosure provides methods for increasing the metabolic rate of 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 pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods for increasing resting energy expenditure in a subject comprising administering to the subject a therapeutically effective amount of compound 1, or a pharmaceutically acceptable salt thereof.

In some embodiments, the subject has at least one of the following: obesity, hyperlipidemia, type 2 diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, atherosclerosis, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, rett's syndrome, metabolic syndrome associated with aging, metabolic diseases associated with increased Reactive Oxygen Species (ROS), friedreich's ataxia, NAFLD, NASH, non-cirrhosis NASH with liver fibrosis, liver steatosis, liver fibrosis, cirrhosis and hepatocellular carcinoma.

In some embodiments, the method comprises increasing the resting metabolic rate without causing a clinically significant risk of adverse events.

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

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

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

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

In some embodiments, the subject experiences an increase in resting energy expenditure of about 30% after administration.

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

In some embodiments, the methods accelerate the natural processes of the human body that improve cardiovascular metabolic processes.

In some embodiments, the subject suffers from 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.

In some embodiments, the subject has a triglyceride blood level above 500 mg/dL.

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

In some embodiments, the subject has refractory hypertriglyceridemia.

In some embodiments, the subject has refractory severe hypertriglyceridemia.

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

In some embodiments, the subject has at least one of abdominal pain, pain in the mid-abdominal, thoracic or back regions, gastrointestinal pain, dyspnea, loss of appetite, nausea, vomiting, pancreatic inflammation, memory loss, dementia, macular tumors, corneal arcs, and xanthomas.

In some embodiments, the subject is an adult male subject.

In some embodiments, the subject is spanish progeny.

In some embodiments, the method comprises reducing low density lipoprotein cholesterol levels and/or reducing non-high density lipoprotein cholesterol levels.

In some embodiments, the method comprises at least one of:

i) Lowering triglyceride levels by at least 5%, at least 10% or at least 20%;

ii) lowering the low density lipoprotein cholesterol level by at least 5%, at least 10% or at least 20%; and

iii) Lowering non-high density lipoprotein cholesterol levels by at least 5%, at least 10% or at least 20%.

In some embodiments, the present disclosure provides a method of reducing liver fat in a subject by at least 50%, comprising administering to the subject a therapeutically effective amount of compound 1, or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods of reducing lipid in a subject by at least 10%, comprising administering to the subject a therapeutically effective amount of compound 1, or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides methods 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 pharmaceutically acceptable salt thereof.

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, gall bladder cancer, head and neck cancer, renal cancer, kidney cancer, leukemia, liver cancer, multiple myeloma, non-hodgkin's lymphoma, ovarian cancer, pancreatic cancer, gastric cancer, and thyroid cancer, and prostate cancer.

In some embodiments, the cancer is associated with obesity, hyperlipidemia, diabetes, hypertension, dyslipidemia, metabolic disease, liver disease, and/or cardiovascular disease.

In some embodiments, the present disclosure provides a method of treating obesity, cancer, hyperlipidemia, 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-cirrhosis NASH with liver fibrosis, liver steatosis, liver cirrhosis, or hepatocellular carcinoma, the method comprising administering to a subject a therapeutically effective amount of compound 1, or a pharmaceutically acceptable salt thereof, so as to achieve at least one of:

ii) a median 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 )；

v) AUC/C of about 18 _max Ratio.

In some embodiments, the method further comprises the steps of: determining the subject before and after administrationVibration Controlled Transient Elastography (VCTE),/>Controlled Attenuation Parameter (CAP) score, magnetic resonance imaging proton density fat fraction (MRI-PDFF), and Enhanced Liver Fibrosis (ELF) score.

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

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

In some embodiments, the subject has an elevated Body Mass Index (BMI).

In some embodiments, the subject has about 28.0kg/m ² To about 45.0kg/m ² Is a BMI of (B).

In some embodiments, the diabetes is type 2 diabetes (T2 DM).

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

In some embodiments, the subject has type 2 diabetes, obesity, HFpEF, HFrEF, NAFLD, and/or NASH.

In some embodiments, the subject has inflammation, fibrosis, cirrhosis of the liver.

In some embodiments, the subject does not experience significant systemic toxicity, serious side effects, adverse events, and/or clinically significant risk of overdosing after administration.

In some embodiments, toxicity, adverse events, and side effects are associated with the granosome decoupling agent.

In some embodiments, the granulometric decoupling agent is 2, 4-dinitrophenol.

In some embodiments, the subject does not experience clinically significant risk of adverse events, side effects, toxicity, and/or overdosing associated with 2, 4-dinitrophenol. Thus, subjects in need thereof can be safely treated without the risk of serious side effects and overdosing.

In some embodiments, the adverse event or side effect includes at least one of nausea, vomiting, sweating, dizziness, headache, cataracts, glaucoma, fever, hyperthermia, tachycardia, sweating, tachypnea, and death.

In some embodiments, the adverse event or side effect includes at least one of fever, hyperthermia, tachycardia, sweating, tachypnea, and death.

In some embodiments, the adverse event or side effect is characterized by at least one of elevated body temperature, accelerated heart rate, abnormal sweating, erythema, perspiration, dehydration, and abnormally rapid respiration.

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

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

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

In some embodiments, the subject experiences saturable absorption of compound 1 such that overdose is prevented. In some embodiments, at higher single doses, saturation of absorption is present. In some embodiments, there is saturation of absorption at a single oral dose of compound 1 above 500mg, above 600mg, above 700mg, above 800mg, above 900mg, above 1000mg, above 1050mg, above 1100mg, above 1200mg, above 1300mg, or above 1400 mg.

In some embodiments, the subject does not experience a correlation between dose and toxicity, adverse events, side effects, or overdosing.

In some embodiments, the clinically significant risk of adverse events, side effects, toxicity, and/or overdosing is prevented by at least one of:

i) Extending half-life (t) of 2, 4-dinitrophenol _1/2 )；

iv) increasing the area under the curve (AUC).

In some embodiments, clinically significant risk of adverse events, side effects, toxicity, and/or overdosing is achieved by providing a maximum plasma concentration of 2, 4-dinitrophenol (C _max ) Is prevented.

In some embodiments, clinically significant risk of adverse events, side effects, toxicity and/or overdosing is provided by administration of compound 1 to provide an average half-life (t _1/2 ) And is prevented.

In some embodiments, the clinically significant risk of adverse events, side effects, toxicity, and/or overdosing is achieved by providing a median time to maximum plasma concentration (T) of 2, 4-dinitrophenol of about 6-8 hours or about 6-10 hours with administration of compound 1 _max ) And is prevented.

In some embodiments, clinically significant risk of adverse events, side effects, toxicity, and/or overdose is provided by administration of compound 1 to provide an extrapolated to infinite median Area Under Curve (AUC) of 2, 4-dinitrophenol of about 3h μg/mL to about 420h μg/mL _inf ) And is prevented.

In some embodiments, a clinically significant risk of adverse events, side effects, toxicity, and/or overdosing is provided by administration of compound 1 to an AUC/C of about 18 _max The ratio is prevented.

In some embodiments, the above methods comprise providing at least one of i) a median time to maximum plasma concentration (T) of compound 1 of about 1-6 hours, about 1-3 hours, or about 1-2 hours _max )；

ii) median half-life (t) of compound 1 of about 1-3 hours or about 1-2 hours _1/2 ) The method comprises the steps of carrying out a first treatment on the surface of the And

iii) Extrapolated to infinite area under median curve (AUC) for compound 1 from about 18h ng/mL to about 380h ng/mL _inf )。

In some embodiments, the subject experiences at least one of:

i) Weight loss of at least 5% or 10%;

ii) a decrease in blood pressure of at least 5mmHg;

iii)HbA _1c a reduction of at least 0.5% or at least 1.5%;

iv) lipid reduction of at least 10%;

v) liver fat reduction of at least 50%;

vi) reduction of serum alanine Aminotransferase (ALT); and

vii) reduction of aspartate Aminotransferase (AST).

Compound 1

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

In some embodiments, compound 1 is administered at about 30mg, 100mg, 200mg, 500mg, or 1050mg per day.

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

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

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

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

In some embodiments, a therapeutically effective amount is selected so as to modulate Cmax, tmax and AUC.

Pharmaceutical salts

The compounds of formula I may be used in their as-is form or as salts. Where it is desired to form a stable non-toxic acid or base salt, it may be appropriate to administer the compound as a pharmaceutically acceptable salt.

Suitable pharmaceutically acceptable salts include those prepared from inorganic and organic acids, including sulfuric acid, 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, pamoic acid (pamoic acid), methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, pantothenic acid, trifluoromethanesulfonic acid, sulfanilic acid, stearic acid, alginic acid, 2-hydroxyethanesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfamic acid, salicylic acid, galactaric acid, β -hydroxybutyric acid, and galacturonic acid; or from ammonium salts and metal salts including calcium, magnesium, potassium, sodium and zinc salts.

Composition/formulation

The pharmaceutical compositions of the present disclosure may be manufactured by processes well known in the art, for example, by means of conventional mixing, dissolving, granulating, dragee-making, suspending, emulsifying, encapsulating, entrapping, lyophilizing processes, or spray drying.

Pharmaceutical compositions for use in accordance with the present disclosure may be formulated in conventional manner using one or more pharmaceutically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. The appropriate formulation depends on the route of administration selected. Pharmaceutically acceptable excipients and carriers are generally known to those skilled in the art and are therefore encompassed in the present disclosure. These excipients and carriers are described, for example, in "Remington' sPharmaceutical Sciences" Mack pub.co., new Jersey (1991).

In some embodiments, the pharmaceutical composition comprises compound 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

In some embodiments, the methods of the present disclosure comprise administering to a subject a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

Pharmaceutical dosage forms

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

In some embodiments, the amount of compound 1 in a 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.

In some embodiments, the amount of compound 1 in a 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.

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.

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

Route of administration

In therapeutic use to control or prevent weight gain in a mammal, the compounds of the present disclosure or pharmaceutical compositions thereof may be administered orally or parenterally.

In certain embodiments, the compounds of the present disclosure or pharmaceutical compositions thereof may be administered orally once daily.

Examples

/>

Example 1: preclinical studies

In the mouse model of NASH (DIAMOND ^TM Mice) and rat models of metabolic syndrome (Zucker diabetic adipose rats), the therapeutic activity of compound 1 was assessed after oral administration. In these models, compound 1 demonstrated efficacy at a dose level of 5 mg/kg/day in mice and at ≡0.5 mg/kg/day in rats.

PK of compound 1 was evaluated in mice, rats and dogs. In all species, compound 1 rapidly absorbed and converted to 2, 4-dinitrophenol, and the results demonstrated that when compared to direct oral administration of 2, 4-dinitrophenol itself, the time to peak plasma concentration (T _max ) Substantially delayed. Figure 1 shows the plasma concentrations of 2, 4-dinitrophenol after administration of compounds 1 and 2, 4-dinitrophenol.

Administration of compound 1 also results in a significantly higher AUC/C of 2, 4-dinitrophenol when compared to direct administration of 2, 4-dinitrophenol _max Ratio. This optimized pharmacokinetic profile of 2, 4-dinitrophenol following oral administration of compound 1 results in substantial improvement of tolerability and safety assessment in animals. In addition, compound 1 was tested for in vitro plasma protein binding in many species and tissue distribution and excretion after oral administration to rats was studied. Assessing the metabolism of compound 1 in vitro using liver microsomes and hepatocytes from non-clinical species and humans, and using human recombinant metabolizing enzymes; these results lead to the selection of rats and dogs as rodent for critical safety tests Dentate animals and non-rodent species. Preliminary investigation of the possibility of drug-drug interactions of compound 1 did not reveal the possibility of drug-drug interactions.

Compound 1 was evaluated in a non-clinical toxicology program including acute toxicity/tolerance, repeat dosing toxicity and genotoxicity studies according to the international coordination committee (ICH) guidelines M3 (R2). All studies critical to support safety assessments were performed in accordance with the Food and Drug Administration (FDA) Good Laboratory Practice (GLP) regulations. In summary, the results of single dose tolerance and repeat dosing toxicity studies in mice, rats and dogs demonstrated that compound 1 was well-tolerated at doses of up to 100 mg/kg/day (7 days), 120 mg/kg/day (up to 63 days) in rats, and 100 mg/kg/day (up to 61 days) in dogs.

There were no compound 1-related deaths or effects on body weight, weight gain, food consumption, body temperature, ophthalmic examination, electrocardiogram, hematology, clinical chemistry, or Urinalysis (UA).

Example 2: single increment dose (SAD) study

Double blind, placebo controlled, phase I studies of the safety and pharmacokinetics of single ascending doses of compound 1 were performed in healthy volunteers. Of a total of 67 subjects (50 actives/17 placebo) in 8 groups, volunteers were orally administered a dose of compound 1 in the range of 30mg once daily (QD) to 1400mg QD. The objectives of this study are as follows:

● After a single dose, a pharmacokinetic profile of compound 1 and 2, 4-dinitrophenol in the plasma of healthy volunteers was established.

● A dose/exposure relationship for a single dose of compound 1 was established over a broad range of exposures in healthy volunteers.

● The pharmacokinetic effect of a 50% fat meal used concurrently with compound 1 was estimated.

● The pharmacokinetics of compound 1 and 2, 4-dinitrophenol are characterized.

Screening procedures occurred from day-28 to day-3. For all groups except group 4 (fed/empty), subjects were allowed to enter the clinical study unit for up to 4 days. Administration of a single dose of study drug occurred on day 1 under fasting conditions (8 hours space-time abdomen), except for the cross-food effect group which had a 10 hour fasting before meal. After all safety assessments and PK analysis blood draws were completed, subjects were discharged on day 3. The fed/empty group remained in the unit and was discharged on day 7.

Double blind dosing occurred in groups 1 to 8. In these groups, six subjects received compound 1 and two received matching placebo. The doses are incremented in the following order: 30. 100, 200, 500, 1400, 1050, and 600mg.

● After pharmacokinetics, group 4 was transitioned to fed state (50% fat meal) administration, and hospitalized patient clearance occurred 72 hours after the first dose and the final dose.

The dose escalation is shown in table 1 below.

Table 1.

Subjects were allowed to enter the clinical study unit on day 1. After 8 hours of time and space, dosing was performed in the morning of day 1, except for the feeding group with 10 hours of empty stomach before meal. Blood draw evaluation PK parameters were performed according to the evaluation schedule. The IC is obtained according to an evaluation schedule. Subjects were released on day 3 after all blood draws and safety evaluations were completed. The fed/empty group remained in the unit and was discharged on day 7.

Schematic study design

Drug administration

Compound 1 or matched placebo was administered orally as a single dose. All subjects except the feeding group were dosed in the morning after 8 hours of time-space and remained in the semi-recumbent position for 1 hour and 4 hours on an empty stomach after administration. The capsule was swallowed with 240mL (8 fluid ounces) of room temperature water.

Group 4 is a cross food effect analysis. Subjects enrolled in group 4 were dosed 30 minutes after a standardized (consisting of 50% fat) meal beginning (after 10 hours of time-space) in order to evaluate food effects. Study drug was administered with 240mL (8 fluid ounces) of water at room temperature.

Studies have shown that the 2, 4-dinitrophenol curves of all groups are extended. By making C _max The curve flattens out providing a "trickle-like" effect for approximately 24/7 delivery, compound 1 maintains the efficacy of 2, 4-dinitrophenol while inhibiting toxicity.

Fig. 2A and 2B show AUC of 2, 4-dinitrophenol after administration of compound 1. It demonstrates the saturable absorption of 2, 4-dinitrophenol after application of compound 1. Thus, overdosing of 2, 4-dinitrophenol can be prevented.

Pharmacokinetics of Compounds 1 and 2, 4-dinitrophenol

Compound 1 was administered in the form of 30mg, 100mg, 200mg, 500mg and 1050mg in the fasting state.

Compound 1 had a median T of 1.75 to 3.00 hours at all dose levels _max And<median T of 0.50 hours _lag And is absorbed rapidly. Average t _1/2 Shorter and in the 500mg and 1050mg fasting groups, in the range of 1.01 hours to 2.32 hours. The average apparent clearance and the volume of distribution appear to increase with increasing dose. Dose-based normalization C _max And AUC, exposure of compound 1 appeared to increase in a dose-proportional manner between the 30mg to 200mg dose groups, and dose proportionality was smaller at doses greater than 200mg (500 mg and 1050 mg).

After administration of compound 1, to<Median T of 0.50 hours _lag And a median T in the range from 6.01 hours to 10 hours _max 2, 4-dinitrophenol appears rapidly. At the dose level, average t _1/2 Relatively long and in the range of 30.0 hours to 38.4 hours. The average apparent clearance and distribution volume was higher for the 500mg and 1050mg dose groups relative to the 30, 100 or 200mg dose groups. Based on C _max And AUC, at 30. Between a dose level of 100 or 200mg and a dose level of 500 or 1050mg, the exposure to 2, 4-dinitrophenol appears to increase in a smaller dose scale, and a 35-fold increase in dose shows an increase in exposure<18 times.

Example 3: assessing food effects

After a single dose of 500mg of compound 1 capsule in the fasting state, a single dose of 500mg of compound 1 capsule was taken together with a standardized (consisting of 50% high fat) meal (after 10 hours of time-space). Geometric mean Compound 1C _max 14.8ng/mL and 25.3ng/mL, respectively, and moderate variability was 52.1% and 49.3% geometric mean CV%. Median T under fasting conditions _max For 3.0 hours and 6.0 hours under fed conditions. Geometric mean AUC _0-24h 53.5h ng/mL and 273h ng/mL, respectively, and high to low variability of 86.5% and 7.92% geometric mean CV%. Geometric mean AUC for fasting and fed groups _last 44.3 ng/mL and 183h ng/mL, respectively, and moderate variability was 66.2% and 44.3% geometric mean CV%.

With respect to C _max Compound 1 geometric mean ratio (90% ci) of 1.82 (1.30-2.56) AUC compared to fasting (reference) _0-24h 5.11 (1.75-14.9), and AUC _last 4.19 (2.75-6.39).

Geometric mean 2, 4-dinitrophenol C _max 694ng/mL and 1680ng/mL, respectively, and moderate variability was 31.6% and 23.3% geometric mean CV%. Median T under fasting conditions _max 8.0 hours and 18.0 hours under fed conditions. Geometric mean AUC _0-24h 12400h ng/mL and 25300h ng/mL, respectively, and moderate to low variability 31.3% and 18.0% geometric mean CV%. Geometric mean AUC for fasting and fed groups _last 34200h ng/mL and 94000h ng/mL, respectively, and moderate to low variability was 46.2% and 25.9% geometric mean CV%.

With respect to C _max The geometric mean ratio (90% CI) of fed (test) to fasting (reference) 2, 4-dinitrophenol is 2.35 (1.85-2.99), AUC _0-24h 2.03 (1.53-2.70), and AUC _last 2.58 (1.88-3.56).

Figures 3a and 3b plasma compound 1 concentrations were compared (linear and semilogarithmic scale) according to food status after oral administration of 500mg compound 1. Figures 4a and 4b below compare plasma 2, 4-dinitrophenol concentrations (linear and semilogarithmic scale) according to food status after oral administration of 500mg of compound 1.

Following administration, a summary of the plasma pharmacokinetic parameters 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.

Cv=coefficient of variation, n=number of subjects with non-missing values, sd=standard deviation,% cv=100× (SD/average), where SD and average are the standard deviation and arithmetic average, respectively, of the untransformed data, geometric% cv=100×sqrt (e ^SD**2 -1), wherein SD is the standard deviation of the natural logarithmic transformation data.

TABLE 2 waiting to continue

Cv=coefficient of variation, n=number of subjects with non-missing valuesFor purposes, sd=standard deviation,% cv=100× (SD/average), where SD and average are the standard deviation and arithmetic average, respectively, of the untransformed data, geometric% cv=100×sqrt (e ^SD**2 -1), wherein SD is the standard deviation of the natural logarithmic transformation data.

TABLE 2 waiting to continue

Cv=coefficient of variation, n=number of subjects with non-missing values, sd=standard deviation,% cv=100× (SD/average), where SD and average are the standard deviation and arithmetic average, respectively, of the untransformed data, geometric% cv=100×sqry (e ^SD**2 -1), wherein SD is the standard deviation of the natural logarithmic transformation data.

Table 3.

TABLE 3 waiting to continue

Cv=coefficient of variation, n=number of subjects with non-missing values, sd=standard deviation,% cv=100× (SD/mean), where SD and mean are the standard deviation and the mean of the untransformed data, respectivelyArithmetic mean, geometric% cv=100×sqrt (e ^SD**2 -1), wherein SD is the standard deviation of the natural logarithmic transformation data.

TABLE 3 waiting to continue

In a full SAD study, compound 1 is rapidly absorbed and converted to a saturatable level of 2, 4-dinitrophenol, which has an average half-life in the range of 1-3 hours. 2, 4-dinitrophenol levels were significantly higher with an average half-life of less than 40 hours. As in the evaluation of the two formulations of compound 1 of different particle sizes, the effect of compound 1 particle size on absorption was evident for positive food effect of compound 1 absorption. At higher single doses, there is evidence of saturation of absorption. AUC/C _max The ratio was about 18, regardless of the dose.

In all subjects in the SAD study, a single oral dose of 30-1400mg of compound 1 was observed to be safe and well tolerated. Serious Adverse Events (SAE) were not reported. There was no demonstrable correlation between dose and incidence of adverse events. The most representative Systemic Organ Category (SOC) of Adverse Events (AEs) was the gastrointestinal tract, consisting mainly of lower abdominal discomfort and thin stool/diarrhea, and higher incidence was observed in compound 1 treated subjects compared to placebo subjects. All AEs were mild or moderate in intensity and most were assigned to mild. Review of ECG, vital signs and laboratory evaluations did not show any trend of abnormal findings or any relationship to dose. Note that some subjects stained urine at higher dose levels, described as green, which appears to be a benign phenomenon.

SAD study demonstrates

● The pharmacokinetics of compound 1 are characterized by rapid absorption and rapid elimination.

● 2, 4-dinitrophenol appears rapidly and has a relatively slow elimination.

● Metabolites form rapidly, have relatively slow elimination, and circulate at low levels (< 1%) compared to 2, 4-dinitrophenol.

● Standard high fat (50%) meals 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. Compound 1 and 2, 4-dinitrophenol Cmax increased > 1.8-fold and compound 1 and 2, 4-dinitrophenol AUC increased > 4.0-fold and > 2.0-fold, respectively, under fed conditions relative to fasting conditions.

Example 4: multiple escalation dose (MAD) study

Multiple escalating doses of safety, pharmacokinetic and pharmacodynamic double-blind, sponsored open, placebo-controlled phase I studies of compound 1 were performed in high MBI volunteers. A first group of 10 healthy higher Body Mass Index (BMI) subjects completed the administration of 200mg of compound 1. The other two groups also planned QD doses of 400 and 550 mg.

The subject well tolerated a 200mg QD dose of oral compound 1. There were no SAE and preliminary evaluation of the data indicated that there were no significant changes in physical examination, vital signs, ECG, or laboratory data. Figure 6 also shows that the 2, 4-dinitrophenol curves for all groups were flat, reaching a steady state of 2, 4-dinitrophenol plasma concentration.

Figure 6 shows that there is no significant increase in body temperature after administration of compound 1.

Figure 7 below shows that compound 1 increased Resting Energy Expenditure (REE) by about 29% compared to placebo.

Figures 8 and 9 show that compound 1 reduced body weight and glucose levels after administration of compound 1.

Figures 10 and 11 show that compound 1 reduces systolic and diastolic blood pressure after administration of compound 1, resulting in a significant reduction in risk of patients suffering from cardiovascular diseases such as HFpFF, hfrref and HFmrEF. The results show that the decrease in blood pressure is statistically very significant and changes are found at all three dose levels. A nearly immediate rapid effect on diastolic and systolic pressure was observed. The decrease in blood pressure and blood glucose and the decrease in obesity strongly indicate that compound 1 is effective in HFpEF. Figure 12 also indicates that a decrease in blood pressure and blood glucose, and a decrease in obesity are not accompanied by an increase in heart rate.

Example 5: phase 2 study of compound 1 in subjects with elevated liver fat and higher BMI

In the presence of elevated liver fat and a higher body mass index (28 to 45kg/m ² ) The 61-day randomized, double-blind, placebo-controlled trial to evaluate the safety and efficacy of the three doses of compound 1 was in progress.

Basic principle of research

High BMI subjects with signs of elevated liver fat better replicate the metabolic profile of the final population of patients with NASH (Golabi et al 2020). Given the forefront of metabolic abnormalities in NASH pathogenesis, in high BMI volunteers with elevated liver fat, understanding the safety of compound 1 and the therapeutic effects on liver fat primarily, and body weight secondarily, would provide greater confidence in the selection of potentially effective doses for phase 2 clinical studies. In a 61 day study, dosages and their associated pharmacokinetic exposures leading to significant reductions in liver fat (greater than or equal to 50% relative to liver fat reduction) were determined, and it was expected that safety, target participation, and efficacy data could be provided to biopsy long term (9 months) 2b in patients with NASH. Furthermore, the clinical effects of granulosa uncoupling were studied in populations reflecting similar metabolic characteristics of obesity (Tainter et al, 1934; harper et al, 2001), providing a further historical safety precedent for the patient population under study.

In summary, understanding the safety and effectiveness of compound 1 on liver fat and body weight in a population of subjects with metabolic characteristics (high BMI and elevated liver fat) of NASH patient populations would provide greater confidence in the dose selection and efficacy of longer term clinical studies following phase 2 b.

Basic principle of selecting a dose

In the SAD study, all single dose exposures of compound 1 and 2, 4-dinitrophenol had acceptable AE profiles. Although compound 1 has a short half-life, the 2, 4-dinitrophenol half-life is about 40 hours. By once daily administration, a significant accumulation of 2, 4-dinitrophenol is expected to be achieved in steady state. According to group 1 of the MAD study, 200mg QDs produced approximately 3.5 fold dose accumulation factor for 14 days, reaching steady state after 7 days of daily dosing.

The pharmacodynamic effects of single dose compound 1 in the SAD study were monitored using Indirect Calorimetry (IC) to determine changes in Resting Metabolic Rate (RMR). In the SAD study, a maximum increase of about 20% in RMR was found in the IC evaluation 33 hours prior to dosing at single dose exposure of compound 1 up to the 1400mg dose.

Given the safety of single dose exposure and the associated pharmacological effects on RMR, it is expected that an initial repeated oral dose of 200mg per day would increase RMR by about 10%. This increase in RMR was demonstrated in group 1 of the MAD study, where an increase in RMR of about 10% was evident to day 7 of 200mg orally daily and continued until day 15 of repeated dosing. In the case of dose accumulation, steady state exposure of 200mg QD dose was as expected. In view of the acceptable safety profile of the 200mg QD dose after 2 weeks of dosing, the next group in the MAD study took 400mg QD of compound 1 for 14 days. The expected increase in RMR was 20% and the dose exposure at steady state was expected at S Within the highest exposure range achieved in the AD study. If the 400mg QD dose had an acceptable safety profile after 2 weeks of dosing, it is expected that the subsequent cohort will receive 550mg QD of compound 1, but the decision will depend on the new data for immediate review. At this predicted dose/exposure, the RMR and expected increase was 30%, well within the impact of dining on RMR, and below the level of acceptable safety profile found by historical clinical studies of 2, 4-dinitrophenol. Given that the half-life of 2, 4-dinitrophenol is about 40 hours, the steady state exposure of 550mg QD compound 1 will be higher than the single dose exposure measured in the SAD study. Population Pharmacokinetic (PPK) models of compound 1 and 2, 4-dinitrophenol concentrations indicate that steady state 2, 4-dinitrophenol concentrations achieved with daily administration of 550mg of compound 1 will still be below the levels observed in clinical and non-clinical studies affecting body temperature. Historically, 2, 4-dinitrophenol concentrations exceeding 28,000ng/mL have been associated with adverse increases in human body temperature (Zhao 2015), and elevated body temperature, wheezing and erythema have been noted in studies conducted in dogs using compound 1, with plasma concentrations of 2, 4-dinitrophenol exceeding 13,000ng/mL. In addition, an increase in RMR of about 200% results in an increase in body temperature (backynsky 2015[ us 201516160 a1 ]). Thus, 550mg QD of compound 1 was administered and C was expected _max And steady state 2, 4-dinitrophenol concentration and resultant increase in RMR is completely lower than 2, 4-dinitrophenol C associated with increased body temperature _max And RMR increases.

The study duration of 61 days was selected based on the duration of administration of the completed compound 1 toxicology study and reflects the clinical experience of using 2, 4-dinitrophenol in 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, and the dose easily resulted in 20-40% increase in RMR. The resulting weight loss is in the range of 1.4-2.1 lbs/week. The drug was well tolerated at these dose levels over the course of 1-3 months. On day 61 of treatment, compound 1 was expected to induce significant liver fat loss, as well as weight loss and dose-related RMR increase in the range of 10-40%. For example, 150mg of compound 1 can increase resting energy expenditure by 10%;300mg of Compound 1 increased resting energy expenditure by 20%; and 450mg of compound 1 increased resting energy expenditure by 30%. Safety and efficacy data from day 61 should provide important guidance for dose selection for long term phase 2b studies.

Main objective

Efficacy:

● The decrease in liver fat content from baseline to day 61 was assessed in subjects treated with compound 1 for elevated BMI compared to placebo, as assessed by magnetic resonance imaging proton density fat fraction (MRI-PDFF).

Safety:

● In overweight and obese subjects defined by BMI, the safety and tolerability of repeated daily dosing of compound 1 for 61 days was assessed.

Secondary target

● The rate and amount of weight loss after 61 days of compound 1 treatment were assessed.

● Changes from baseline in systemic obesity were assessed by MRI 61 days after compound 1 treatment.

● In high BMI subjects, PK curves for Compound 1 and 2, 4-dinitrophenol were characterized 61 days after dosing.

● After compound 1 administration, the change in liver component measurements from baseline and the change in liver fat content were assessed and correlated.

● The Pharmacodynamic (PD) effect of compound 1 on metabolic and cardiovascular risk factors was studied.

● The PD effect of compound 1 on metabonomics, proteomics and lipidomic features was studied.

● The efficacy of compound 1 and the dose/exposure relationship of PD effect were characterized, where the data permits.

Endpoint (endpoint)

Primary endpoint

● Relative change from baseline in liver fat content assessed by MRI-PDFF on day 61.

Secondary endpoint

Pharmacodynamics of medicine

● Day 61 change in body weight from baseline.

● Changes from baseline in systemic obesity on day 61.

● Changes from baseline in surrogate indicators of liver inflammation and fibrosis on day 61:vibration Controlled Transient Elastography (VCTE),/>Controlled Attenuation Parameter (CAP) score and Enhanced Liver Fibrosis (ELF) score.

● Changes in lipid parameters and cardiovascular risk biomarkers from baseline on day 61: serum hypersensitive 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 in metabolic disease parameters from baseline on day 61: steady state model assessment of insulin resistance (HOMA-IR), fasting blood glucose concentration, glycated albumin concentration, hbA _1c 。

Safety and tolerability

● Summary of physical examination results during the study.

● Adverse Event (AE) evaluation during the study.

● Vital sign parameters were evaluated during the study. Parameters include resting systolic and diastolic pressures, resting heart rate, resting respiratory rate, and oral body temperature.

● Body weight was assessed during the course of the study.

● Safety 12 lead ECG during study.

● Clinical laboratory values (hematology, whole biochemical group (including lipidome, CPK, magnesium, liver function test) and Urine Analysis (UA) were assessed during the 61 day dosing period.

● Ophthalmic examinations, including slit lamps, were evaluated before and after 61 days of dosing.

Pharmacokinetics of

● Compound 1 and 2, 4-dinitrobenzenePopulation PK analysis of phenol. The following PK parameters will be estimated as appropriate: c (C) _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 where data allows and appropriate.

● As the data allow, a non-compartmental analysis was performed on compound 1 and 2, 4-dinitrophenol. The following PK parameters will be calculated: c (C) _max AUC and accumulation.

● The exposure response relationship of compound 1 and 2, 4-dinitrophenol and efficacy/pharmacodynamic endpoints was modeled as appropriate.

Exploratory endpoint

● Changes from baseline in metabolomics and lipidomic characteristics (One Way river- [ OWL ] metabolomics and lipidomic analysis) at day 61.

● Proteomic profiling at day 61Change from baseline.

● As the data allow, serum alanine Aminotransferase (ALT) and aspartate Aminotransferase (AST) concentrations changed from baseline on day 61.

Study design

This is a phase 2a, randomized, parallel group, placebo-controlled, double-blind, repeat dose study, in subjects with high BMI and evidence of elevated liver fat, safety and efficacy of three oral dose levels of compound 1 compared to placebo were assessed over the course of 61 days, as shown in the following figure:

10 visits, screens, baseline/first dose, day 2, day 7, day 14, day 28, day 42,

Day 49, day 61/end of treatment, and follow-up on days 71-75.

Access procedures included PK and PD measurements and security assessments.

Subjects will be screened over a period of 45 days to determine their qualifications based on specific medical history, physical, laboratory, and imaging evaluations. Due to the programming, multiple visits may be required to complete the screening process. However, if all screening evaluations and procedures (including MRI) can be completed within 30 days of the first administration, one screening visit is allowed.

Once qualified, the patients will be randomly assigned to one of the compound 1 treated group or matched placebo control group, administered once daily (fasting) for a total of 61 days. During the 61 day dosing period, the subjects will return to the clinic for frequent assessment visits. Follow-up will be performed within 10 to 14 days after dosing is complete.

Subjects will be instructed to maintain the same diet and activity/exercise level throughout the study as before participating in the study.

The administration is once daily in a fasting state. Eligible subjects will be randomly assigned (n=20 per group) to 1 out of 4 treatment groups:

● Group 1: the placebo was administered orally once daily for 61 days.

● Group 2: 150mg of Compound 1 was orally administered once daily for 61 days.

● Group 3: 300mg of Compound 1 was orally administered once daily for 61 days.

● Group 4: 450mg of Compound 1 was orally administered once daily for 61 days.

Will prevent randomization and according to HbA _1c Layering (normal range and between 5.7% and 9.0% (including 5.7% and 9.0%)).

Inclusion criterion

The subject must meet all of the following inclusion criteria to qualify:

1. adult males or females, with informed consent, are aged 28 to 65 years (inclusive) and BMI 28.0 to 45.0kg/m ² And (containing) the same.

a. Female subjects with fertility must be non-lactating, negative by urine pregnancy test at screening to confirm non-pregnancy, and agree to continue to use the effective contraceptive method for at least 4 weeks or barrier method for 2 weeks prior to the first study drug administration until 30 days after the last study drug.

b. Female subjects with fertility must not donate eggs during the study period and at least 30 days after the last dose of study medication.

c. Female subjects who are not fertility must be surgically sterilized (e.g., hysterectomy, bilateral tubal ligation, ovariectomy) or postmenopausal (no menstruation >1 year at screening, and Follicle Stimulating Hormone (FSH) > 40U/L).

d. Male subjects who have not undergone a vasectomy and/or subjects who have undergone a vasectomy but who have not undergone 2 post-operative sperm tests are negative must agree that acceptable contraceptive methods are used 30 days after the first dose of study medication to the last dose of study medication and that sperm are not donated for at least 30 days during the study period and after the last dose of study medication.

2. This can be incorporated based on the investigator's assessment of general medical condition and recorded by medical history, physical examination, vital sign assessment, 12-lead ECG, clinical laboratory assessment, and general observations.

a. The subject must take a stable dose of the drug at least 2 months prior to screening to treat the underlying obesity-related disease.

b. Diabetics may be treated with metformin, DPP-4 inhibitors or sulfonylureas, but must take a steady dose at least 2 months prior to screening.

c. At the time of screening, if commensurate with potential obesity or related metabolic dysfunction (e.g., dyslipidemia and hyperglycemia) in a qualified subject, certain laboratory values may be outside of reference range unless such abnormalities indicate potential conditions that may affect the safety of the test subject or interfere with the evaluation of compound 1 or affect the interpretation of the study results.

d. Other assessments deemed clinically significant by the researcher (clinical laboratory tests, ECG, vital signs, physical examination) are repeated at the discretion of the researcher for abnormalities or deviations outside of normal ranges. Results that continue beyond the normal range must be judged by the researcher to be clinically insignificant and acceptable for research participation.

e. Subjects with elevated unbound bilirubin are allowed due to the presumed gilbert syndrome.

f. The subject must be thyroid-normal as assessed by thyroid profile using Thyroid Stimulating Hormone (TSH) and free thyroxine (T4) tests at the time of screening. Subjects with a history of stable thyroid disease who have taken a stable dose of thyroid medication for at least 4 months may be enrolled in the group.

3.CAP scoring>300dB/m。

MRI-PDFF showed > 8% liver fat.

5. Knowing the procedure and requirements of the study and providing written informed consent and authorizing disclosure of the protected health information.

6. Is willing and able to comply with the requirements of the research scheme.

Exclusion criterion

Subjects will be excluded from the study if any of the following criteria are met:

1. insulin-controlled diabetes.

2. Pregnancy or lactation or planned pregnancy.

3. Intolerance to Magnetic Resonance Imaging (MRI) or MRI procedure contraindications exist, including but not limited to inability to fit into an MRI scanner or having surgical clips/metallic implants/clips. The subject must not have claustrophobia, have history of claustrophobia or be intolerant of confined or small spaces.

4. Weight gain or loss >5% within 3 months prior to study or >10% within 6 months prior to screening.

5. Screening for a history of gastric banding, intragastric balloon, duodenal-jejunal cannulas, or bariatric surgery within 5 years, planning bariatric surgery prior to the end of study participation, or planning weight loss during the study by special diet, exercise program, or both.

6. History of hyperthermia.

7. A history of chronically severe recurrent rashes of unknown cause.

8. Cardiovascular diseases of past or current clinical significance, including but not limited to transient ischemic attacks, strokes, arrhythmias, syncopes, unstable angina, myocardial infarction within 6 months prior to screening, congestive heart failure, or uncontrolled hypertension. (uncontrolled hypertension is defined as systolic pressure. Gtoreq.160 mmHg or diastolic pressure. Gtoreq.100 mmHg, based on the average of three resting measurements using appropriately sized cuffs in the sitting position).

9. Resting heart rate <45 or >110bpm.

10. Screening ECG or based on medical history:

qt/QTcF interval is significantly prolonged by baseline (e.g., men repeatedly show QTcF intervals >450 ms, women >470 ms).

b. Other risk factor medical history of torsades de pointes (TdP) such as heart failure, hypokalemia, family history of long QT syndrome, or family history of sudden cardiac death of unknown cause.

11. Kidney disease, kidney transplantation or estimation of glomerular filtration rate (eGFR) based on CKD-EPI creatinine equation (NKF 2009; https:// www.kidney.org/content/CKD-EPI-creatine-equation-2009)<50mL/min/1.73m ² 。

12. Serious pulmonary diseases requiring chronic daily dosing include Chronic Obstructive Pulmonary Disease (COPD), emphysema, pulmonary fibrosis or asthma.

13. Untreated Obesity Hypoventilation Syndrome (OHS) or Obstructive Sleep Apnea (OSA).

14. Non-alcoholic fatty liver disease (NAFLD)/non-alcoholic steatohepatitis (NASH) other than past or active (acute or chronic) liver disease, such as, but not limited to, autoimmune liver disease, viral hepatitis, hereditary hemoglobinopathy, primary biliary cirrhosis, wilson's disease, alpha-1-antitrypsin deficiency, alcoholic liver disease, gestational acute fatty liver, or drug-induced (including acetaminophen) liver disease.

15. Clinically significant history or treatment of gastroparesis, inflammatory bowel disease, or any upper gastrointestinal surgery other than cholecystectomy, or small gastric surgery approved by a medical inspector.

16. History of cirrhosis and/or liver function decompensation, including ascites, hepatic encephalopathy or variceal bleeding.

17. Screening for a history of acute pancreatitis or chronic pancreatitis of any cause within one year.

18. Serum triglyceride concentrations were above 500mg/dL.

19.HbA _1c >9.0％。

20. Family (mother/father/sibling) and/or history of personal retinal detachment at any time in the past.

21. Any history or current diagnosis of glaucoma.

22. The following evidence of ophthalmic examination at the time of screening:

a. peripheral retinopathy in need of treatment, retinal tears, or lattices in need of treatment.

b. Optical Coherence Tomography (OCT) and examination revealed diabetic retinopathy with macular exudates or macular edema.

c. Any active macular disease that affects vision, including macular pucker (epiretinal membrane) and macular degeneration.

d. Visually significant cataracts as determined by an ophthalmic physician.

e. Any previous intravitreal injection of anti-VEGF drug treats macular degeneration.

f. History of past vitrectomy.

23. Screening for a history of malignancy within 5 years, except basal cell or squamous cell skin cancer, cervical cancer in situ, or prostate cancer, currently or is not expected to require radiation therapy, chemotherapy, and/or surgical intervention or initiation of hormonal therapy.

24. History of organ transplantation.

25. Less than 1 week prior to dosing (visit 2/day 1) and/or the vaccine of covd-19 was scheduled for the duration of the study.

26. Screening for a history of severe drug abuse in the previous year, or for frequent use of soft drugs (e.g., cannabis) in the 3 months prior to the screening visit, or for hard drugs (e.g., cocaine, phencyclidine [ PCP ], opioid derivatives including heroin, and amphetamine derivatives) in the previous 1 year.

27. There was a history of alcohol abuse over the past 2 years, or by screening using the drinking disorder identification test (audiot, thompson 2018), there was evidence of excessive drinking at present, and a history of frequent drinking of 14 cups per week and 7 cups per week (1 cup = 4 oz (120 ml) wine or 12 oz (360 ml) beer or 1 oz (30 ml) spirits per female) within 6 months of screening, as determined by the investigator.

28. Urine drug screening for drug abuse is positive or phosphatidyl ethanol (PEth) blood test results are positive >100ng/mL. In the case of exclusionary PEth values, groups may be considered if the primary investigator and medical inspector agree that the subject's medical history is inconsistent with alcohol abuse.

29. At present, the cigarettes are regularly smoked or the cigarettes with more than 5 cigarettes or the same number are smoked every week. Allowing the use of nicotine patches for smoking cessation.

30. The detection result of the hepatitis B surface antigen (HBsAg), the hepatitis C virus antibody (HCV Ab) or the human immunodeficiency virus (HIV 1/2) antibody is positive.

31. Neutropenia is defined as absolute neutrophil count < 1000/. Mu.L.

32. Serum AST or ALT >5x normal Upper Limit (ULN) at the time of screening. (at the discretion of the investigator, one repeat test may be allowed to proceed within 7 days).

33. Total bilirubin > ULN is considered normal variability without other clinically relevant liver injury unless approved by a medical supervisor or due to gilbert syndrome.

34. If there is evidence of other potentially significant liver damage, the international normalized ratio at screening (INR) is 1.3 or more.

35. Another clinical trial was taken at screening or exposure to any study drug, including topical, was performed within 30 days of screening or within 5 half-lives if half-life is known.

36. There were no tattoos or body perforations during the study. Researchers or sponsors consider any potential physical or psychological medical condition that would make it unlikely that a subject would be following the requirements of a study or would not be able to complete a study.

37. The researcher considers any situation that would interfere with his/her ability to provide written informed consent, follow the instructions of the study, or possibly confuse the interpretation of the study results or put the subject at undue risk.

38. Known or potential hypersensitivity to compound 1 or an excipient thereof.

Disable drug (currently used):

39. any herbal supplement, over-the-counter, mail order or prescription for weight loss.

40. Prescription or over-the-counter stimulants include: dextroamphetamine/dextroamphetamine, dextroamphetamine/amphetamine combination/aderall or methylphenidate

41. Thiazolidinediones (TZDs): pioglitazone/Actos, rosiglitazone/Avandia.

42. Glucagon-like peptide 1 (GLP 1) agonists: exenatide/byuta/Bydureon, lixisenatide/Adlyxin, liraglutide/Victoza, dulcitide/Trulicity, cord Ma Lutai (semaglutide)/Ozempic.

43. Sodium-glucose cotransporter 2 (SGLT 2) inhibitors: canagliflozin (canagliflozin)/Invokana, dapagliflozin (dapagliflozin)/Farxiga, eagliflozin (empagliflozin)/Jardiancene, ertuglifuzin/Steglatro.

44. Vitamin E: xiong Erchun (ursodiol) or high dose vitamin E >400 IU/day for at least 1 month in the last 6 months, or high dose vitamin E is started in the last 3 months of screening.

45. Recently (within 3 months of screening) or currently using obeticholic acid/ocauliva, systemic corticosteroids, methotrexate, tamoxifen, amiodarone or long term tetracyclines.

46. Warfarin, heparin, factor Xa inhibitors (dabigatran), betrixaban (betrixaban), ai Duosha shift (edoxaban), apixaban (apixaban) and rivaroxaban (rivaroxaban)).

47. The prolonged QT/QTc interval, as determined in https:// credullemeds. Org/website list category "known risk" and concomitant medications associated with increased risk of torsades de pointes are known.

48. Products containing Cannabidiol (CBD).

Treatment planning and method

The study procedure should be completed as specified in the evaluation schedule (table 4).

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Pharmacokinetic parameters

The following PK parameters will be determined from the concentration-time data:

● Group PK (PPK) analysis of compound 1 and 2, 4-dinitrophenol. The following PK parameters will be calculated: c (C) _max 、T _max 、t _1/2 、T _lag 、AUC _0-t 、AUC _0-∞、 CL/F, vd/F, λz. Other PK parameters may be calculated where data allows and appropriate.

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

Magnetic resonance imaging proton density fat fraction (MRI-PDFF) is a non-invasive, quantitative biomarker for assessing liver fat content (steatosis). At baseline and end of treatment, the percentage of fat or Proton Density Fat Fraction (PDFF) in the liver was measured using MR: MRI-PDFF. The liver volume will be estimated from an axial T1 weighted or dual echo gradient echo image covering the whole liver. This advanced MRI technique measures the mobile proton fraction in the liver attributable to liver fat (PDFF), which is a direct measure of liver fat content and is a fundamental tissue property. Subjects were required to have a 4 hour empty stomach prior to performing MRI-PDFF.

Abdominal MRI

MRI images of the abdominal region will be taken to assess total fat. The two main measurements of this scan will estimate the total Visceral Adipose Tissue (VAT), i.e. the fat type stored in the body cavity, and the Subcutaneous Adipose Tissue (SAT), i.e. the fat type visible under the skin.

As a non-invasive medical device, liver fat content (steatosis) and liver hardness (fibrosis) can be estimated. The measurement works by measuring shear wave velocity. The 50-MHz wave is transmitted from a small transducer at the end of the ultrasound probe to the liver. The probe tip also has a transducer for measuring the velocity of the shear wave (in meters per second) as the wave passes through the liver. Shear wave velocity translates to liver stiffness, which is expressed in kilopascals (VCTE scores). A second measurement is also made which estimates liver steatosis by measuring the ultrasound attenuation of echoes, known as Controlled Attenuation Parameters (CAPs). In progress Previously, the subjectIt takes 4 hours to empty.

One Way river assay (OWL)

OWL-metabonomics

This metabonomics assay extracts metabolites from volunteer plasma and serum to take a snapshot of cell function. Liquid chromatography-mass spectrometry (LC-MS) metabonomics was used to identify serum biomarkers that distinguish normal liver from NAFLD and NASH from NAFLD. Metabolomic profiling also provides insight into cellular function and inflammation by examining various cellular metabolites that provide insight into key molecular pathways.

OWL-lipidomics

Lipidomics are non-invasive blood assays that analyze and recognize lipids in plasma and serum. These lipids will be isolated and characterized by mass spectrometry and analysis will include fatty acids, fatty acid derivatives, glycerolipids, glycerophospholipids, sphingolipids and sterols.

Slow dissociation rate modified aptamer (SomaScan)

SomaScan is a non-invasive blood assay. Plasma and serum samples will be analyzed using oligonucleotide aptamers whose three-dimensional conformational shape specifically binds to a protein target of interest. Over seven thousand proteins will be analyzed and quantified to achieve a proteomic snapshot of the body. Different mathematical models have been applied to large clinical data to build algorithms with predictive values in terms of cardiovascular health, metabolic rate, lean body mass, liver inflammation, cardiopulmonary health and glucose tolerance.

Enhanced Liver Fibrosis (ELF)

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 matrix metalloproteinase 1 tissue inhibitor (TIMP-1). The values of these three markers, when used in combination with accompanying clinical data, can highly predict the inflammatory and fibrotic status of the liver as demonstrated by correlating data with histology in larger clinical trials.

Exploratory biomarkers

Blood samples were obtained and separated into plasma and serum fractions from which individual 400 μl aliquots were extracted, labeled and stored for future analysis of protein, lipid or gene expression to help explain the pharmacological effects of compound 1.

Ophthalmic examination

A comprehensive ophthalmic examination will be performed at the completion of screening and dosing (about day 61), including fundus photographs of the posterior poles of the eyes, OCT and slit lamp evaluation of the macula of both eyes, to characterize the baseline status of the subject and monitor any changes from baseline during treatment. Pupil dilation will be accomplished using 2.5% new synephrine and 0.5% topiramate (Mydriacyl) (unless contraindicated by the ophthalmologist), once in light eyes, one drop per eye, and up to two times in dark eyes, 5 minutes apart. Slit lamps are biological microscopes that use intense light in ophthalmic examinations to evaluate different structures in the front of the eye and the interior of the eye to determine health and detect eye disease. OCT is a non-invasive imaging technique that uses light waves to take a cross-sectional photograph of the retina. Fundus photography involves photographing the back of the eye. These procedures are standard tests involving eye health medical assessment, which should be performed by a limited number of coordinated ophthalmologists to ensure consistency of the assessment.

Vital signs

Vital signs include body temperature, systolic and diastolic blood pressure, heart rate and respiratory rate. All blood pressure readings must be taken using a blood pressure cuff that fits the size of the subject's arm. Too small a blood pressure cuff may result in inaccurate, higher blood pressure measurements. Blood pressure and heart rate were recorded after 5 minutes or more supine on study subjects. Three blood pressures were obtained at each time point, each blood pressure being obtained approximately 2 minutes apart. The first blood pressure will be discarded. The second and third blood pressure measurements will be entered into the database, averaged, and will be the value at that time point.

The InBody scale will be used to measure body weight, muscle and body fat. This must be done before administration, in a fasting state, and at about the same time of day.

Subjects will also monitor body temperature daily at home using a Braun Thermoscan 7 inner ear thermometer provided to them. The thermometer provides a color coded display screen that displays temperature as well as normal, elevated (> 99.9°f, yellow display) or exothermic (> 103°f, red display) temperatures. There is an acoustic feedback system that ensures proper use and alerts the user that a temperature has been obtained. The first nine readings will be recorded on the thermometer. Temperature readings should be taken daily for administration. If the subjects have symptoms that may indicate fever, they should measure body temperature and verify. For a temperature rise of ≡100°f, as shown by the yellow or red color, the subject should stop taking compound 1, avoid using antipyretics (acetaminophen, aspirin or a non-steroidal anti-inflammatory drug) and call the research center.

Clinical laboratory testing

Only at screening:

● Viral serology will involve the detection of the presence of hepatitis b antigens, anti-hepatitis c antibodies and anti-HIV antibodies.

● TSH and free T4.

● eGFR=CKD-EPI creatinine equation (NKF 2009; www.kidney.org/content/CKD-EPI-creatine-equence-2009).

Hematological tests will include the mean red blood cell hemoglobin concentration (MCHC), mean red blood cell volume (MCV), hematocrit, hemoglobin, white blood cell count, absolute counts of lymphocytes, monocytes, neutrophils, basophils, eosinophils, and platelets.

Serum chemistry analysis will include 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-glutamyl transferase (GGT), bilirubin (total and direct bilirubin), amylase, and CPK.

● Baseline ALT and AST = although each value will be recorded, the mean of the screening and day 1 pre-dose values will be used as the baseline value in the statistical evaluation of these parameters in the exploratory analysis.

The lipidosome group will include total cholesterol, HDL, LDL, VLDL, triglycerides and FFA.

Other tests performed at selected time points include glycated albumin, hs-CRP, apoB, lp (a) and HOMA-IR (including glucose, insulin and C-peptide; wallace 2004) and PEth tests.

● For HOMA-IR,3 blood samples of 3 analytes (blood glucose, serum insulin and C-peptide) will be drawn after at least 5 minutes of separation between each sample.

● The PEth test is a serum biomarker that can be assessed for recent alcohol consumption. This value depends on the amount of alcohol consumed and the time after consumption. This will be evaluated at screening and day 28. This value may be obtained at other times at the discretion of the researcher if there is concern about excessive drinking based on medical history, symptoms or laboratory assessment (e.g., elevated liver function tests).

Urine analysis will include a dipstick evaluation, which is performed if the dipstick shows less (1+), medium (2+) or greater (3+), blood or protein. If urine > trace protein in 2 collections, urine protein and albumin need to be detected.

Female subjects will be subjected to a urine pregnancy test.

Laboratory tests may be repeated once during screening. In assessing adverse events, researchers may make additional laboratory assessments as appropriate and according to medical needs.

12-lead electrocardiogram

After the subject has rested in the supine position for at least 10 minutes, a single 12-lead ECG measurement will be obtained. External stimuli should be kept to a minimum. During this time, video games, television watching, and talking are not allowed. The test will use a digital ECG machine. If the ECG time point is consistent with any blood sample, the ECG will be performed within 10 minutes of obtaining the blood sample at the same time point. Furthermore, the subject should not eat within 2 hours prior to ECG administration, as long as possible.

The ECG will be measured using an ECG machine that automatically calculates heart rate and measures PR, RR, QRS, QT and QTcF (friericia correction formula). The same ECG machine should be used for the same subject throughout the study if possible. The ECG should be performed in accordance with the sponsor acceptable study entity SOP.

Example 6: by usingCompounds for treating obese subjects with heart failure (HFpEF) retaining ejection fraction Exploratory phase 2A study of 1

This is a phase 2A, randomized, parallel group, placebo controlled, double blind, in-subject dose escalation trial with 3 dose levels of compound 1 and placebo. It is expected that 62 participants will be in the group. The subjects will receive either compound 1 or placebo at random (1:1). The two dose levels will be administered sequentially (150 mg per day, then 300mg per day), each for 20 days, reaching the third and highest dose of 450mg per day if the lower previous 2 doses demonstrate safety and tolerability. The administration of a high dose of 450mg will last for a total of 94 days, with a safety follow-up within-14 days of the last administration.

Subjects will be screened over a 40 day period according to an evaluation schedule to determine their qualification based on specific medical history, physical, laboratory and imaging evaluation. Although one screening clinical site visit is required, additional visits may be required to complete the screening procedure due to scheduling problems. Some of these assessments will be taken as a baseline prior to dosing. The central laboratory will be used for all evaluations including MRI, DEXA, clinical blood/plasma measurements, transthoracic echocardiography, and CPET.

Compound 1 was evaluated for efficacy in improving cardiovascular function in HF (HFpEF) obese subjects with preserved ejection fraction.

Inclusion criteria:

1. adult males or females, are no less than 40 years old.

2. Information provided in the Institutional Review Board (IRB) or independent ethics board (IEC) approved Informed Consent Form (ICF) can be understood and the form must be signed before any study procedure is initiated.

3. Body Mass Index (BMI) >30kg/m2;

4. at the discretion of the investigator, the signs and symptoms of HF were as follows:

kccq OSS <80; NYHA class II-III; c. the baseline peak VO2 is less than 18mL/kg/min for females or less than 20mL/kg/min for males; d. respiratory exchange ratio at baseline (respiratory quotient) (RER [ RQ ]) >1.0; e. left ventricular Ejection Fraction (EF) >50%; f. at least 1 of the following HF objective criteria are satisfied: there has been a record in the last year that hospitalization was mainly due to HF, or if more than one year, an increase in structural heart disease (left atrial volume increase or left ventricular hypertrophy with gender specific tangent points according to Lang, 2015) on echocardiography was as follows:

■ Left Ventricular Hypertrophy (LVH):

a. male: the thickness (cm) of the partition wall is >1.1 or the thickness of the rear wall is >1.1;

b. female: the thickness (cm) of the partition wall is >1.0 or the thickness of the rear wall is >1.0;

■ Left atrial expansion (LAD): AP size (cm): male >4.0; female >3.8; in the last year, pulmonary Capillary Wedge Pressure (PCWP) at rest >15mmHg (or left ventricular end-diastole pressure [ LVEDP ] >18 mmHg) or at exercise >25mmHg (or 2.0 mmHg/L/min); in doppler and tissue doppler imaging in recent years, the E/E' ratio of the resting time-cell spacer ring is >14; in the absence of atrial tremor/flutter, the present elevated NT-proBNP is defined as >125pg/mL, and for subjects with atrial tremor/flutter >375pg/mL.

5. Participants should maintain a steady physical activity level throughout the study and must agree not to participate in the athletic training program during the study.

6. Participants should remain on a steady diet and not be intended to participate in weight loss programs prior to or during the course of the study.

7. Thyroid function is normal as assessed by thyroid profile using Thyroid Stimulating Hormone (TSH) and free thyroxine (T4) tests at the time of screening. Subjects with a history of stable thyroid disease who have taken a stable dose of thyroid medication for at least 4 months may be enrolled in the group.

8. Ambulatory (wheelchair-independent or scooter-independent) and capable of upright exercise testing, including 6MWT.

9. Drug dose stabilization 30 days prior to screening (defined as no new drug or no change in existing drug dose > 50%) and other specific criteria for diuretics:

a. if a loop diuretic or a thiazide diuretic is used for treatment, a stable treatment regimen must be employed, which allows for a flexible diuretic dosing regimen.

Exclusion criteria:

1. at the discretion of the investigator, life expectancy was <1 year for non-cardiovascular reasons.

Within 2.5 years there was a history of malignancy (except for non-high grade skin cancer, carcinoma in situ, or low grade prostate cancer).

3. Weight change (increase or decrease) was ≡10 pounds by self-report or weight loss recorded over the past 90 days.

4. Bariatric surgery prior to screening or during research is planned.

5. Screening treatment with GLP-1 receptor antagonists began within 1 year.

6. Treatment with SGLT2 inhibitors was started within 6 months of screening.

7. Intolerance to MRI or MRI procedure contraindications exist, including but not limited to:

a. there are surgical clips/metal implants/shrapnel/internal motorized implants; or alternatively

b. The subject is accustomed to or exceeds the weight tolerance limit of the scanner (typically 350 or 400 pounds, depending on the manufacturer) and cannot fit into the MRI scanner; or alternatively

c. Claustrophobia). Can lead to a history of severe claustrophobia where MRI is not possible.

8. Current acute decompensated HF requires Intravenous (IV) diuretics or recent (< 1 month prior to screening) hospitalization for HF.

9. Primary cardiomyopathy (e.g., constrictive, restrictive, invasive, toxic, hypertrophic [ congenital ], congenital, or any other primary cardiomyopathy at the discretion of the researcher.

10. Active myocarditis (covd induced or otherwise).

11. Active collagen vascular disease.

12. Researchers believe that there are currently greater than moderate left or right valve disease.

13. During participation in the trial, cardiac surgery or catheter intervention was planned.

14. In the last 3 years, previously recorded EF <40%.

15. Tachycardia (> 110/min) at screening.

16. Atrial tremor or atrial flutter, uncontrolled heart rate response, or ECG shows resting heart rate greater than 110bpm at screening. The subject may be rescreened after appropriate adjustments to the medication to control atrial fibrillation. Up to 16 subjects with this condition can be included in the study.

17. Untreated, life threatening arrhythmias.

Results of phase 2a test

The 2a phase metabolism test of compound 1 was a 61 day randomized, double-blind, placebo-controlled test aimed at assessing three dosage levels of compound 1 (150 mg, 300mg and 450 mg) in obese participants (body mass index 28 to 45 kg/m) with elevated liver fat (greater than 8%) ² ) Safety and effectiveness of the system. Eighty (80) participants between 28 and 65 years of age were randomized to one of three compound 1 treatment groups or matched placebo groups, stratified and blocked HbA1C levels of 5.7% or higher, and dosed once daily (fasting). Indicating that the participant is not to change the behavior in terms of diet or exercise. Phase 2a trials reached primary (reduction of liver fat by MRI-PDFF) and secondary (reduction of body weight and fat by abdominal MRI) endpoints. Key results and observations include:

● At all three dose levels, reduction of liver fat was statistically significant (p <0.0001 for ANCOVA).

The relative reductions in liver fat were 33%, 43% and 40%, corresponding to 40%, 71% and 72% response rates (> 30% relative reduction) at low, medium and high doses, respectively. Placebo relative reduction of liver fat was 2% and response rate was 5%.

● The main cardiovascular and metabolic health indicators observed at all dose levels included:

dose-dependent decrease of glycated albumin, which is an indicator of glucose control and insulin function (p <0.0001, high dose).

Dose-dependent decrease of the inflammatory marker hypersensitive C-reactive protein (hsCRP), an important parameter of cardiovascular risk (p <0.005, high dose).

● Compound 1 was well tolerated at all dose levels with excellent compliance. No serious adverse events or deaths were reported. Diarrhea and transient flushes associated with alcohol intake occurred in 25% and 31.6% of compound 1 subjects, respectively, as the most commonly reported adverse events occurring during treatment. Most of these events are mild; one participant discontinued compound 1 due to diarrhea in the low dose group, while no participant discontinued for any reason in the high dose group.

The specific efficacy and safety of the phase 2a results are shown in the following figures and tables:

(1) As shown in table 5, compound 1 was well tolerated for eight weeks.

Table 5.

^a Adverse events associated with treatment are those that the researcher evaluates as likely or likely to be associated with the research treatment

(2) The body temperature changes are shown in fig. 13 and table 6.

Table 6 study day


				Fixation Effect (type III)	P value	P value summary	Is there a statistical significance (P < 0.05)?
Time	0.1543	ns	Whether or not
				Treatment of	0.4318	ns	Whether or not
Time x treatment	0.7274	ns	Whether or not

(3) PK results (mean ± SEM) are shown in figure 14.

(4) As shown in fig. 15 and table 7, all doses showed therapeutic effect.

The MRI Proton Density Fat Fraction (PDFF) was corrected for percent change from placebo at baseline.

Table 7.

Baseline was the last non-missing value before the first dose of study drug

(5) All doses showed therapeutic effects as shown in fig. 16 and table 8.

Covariance analysis of MRI Proton Density Fat Fraction (PDFF). Mean change from baseline on day 61 of FAS population (LS mean ± 95%)

TABLE 8

Note that: this analysis was performed using a model that included treatment and baseline HbA1c stratification as factors as fixed effects, and baseline values of the response variables as covariates

Note that: ls=least squares, ci=confidence interval

(6) The therapeutic results in response (MRI-PDFF liver fat reduction > 30%) for all dose groups are shown in table 9.

TABLE 9

On day 61, MRI-PDFF response rates of 450mG and 300mG were comparable at > 300mG

(7) As shown in fig. 17 and table 10, compound 1 doses of 300mg and 450mg significantly reduced body weight once a day.

Repeated measures of InBody body weight analysis. Mean change from baseline (LS mean ± 95% ci) for FAS population.

Table 10

(8) The body weight observation results are shown in fig. 18 and table 11.

TABLE 11


						Day 61
Placebo compared to 150mg	1.131	-1.302 to 3.564	Whether or not	ns	0.4362
						Placebo compared to 300mg	3.904	1.665 to 6.142	Is that	**	0.0057
Placebo compared to 450mg	6.014	3.820 to 8.209	Is that	****	＜0.0001

(9) Visceral adipose tissue observations of FAS populations are shown in fig. 19 and table 12.

The treatment group FAS population had a placebo corrected percent change (mean ± SEM) in abdominal MRI liver volume and obesity from baseline.

Table 12

(10) Subcutaneous adipose tissue observations of FAS populations are shown in fig. 20 and table 13.

TABLE 13

(11) Fat loss (total adipose tissue) was confirmed by MRI, as shown in fig. 21.

(12) Liver volume observations of FAS populations are shown in fig. 22 and table 14.

TABLE 14

(13) The observed systolic blood pressure is shown in figure 23.

Mean change from baseline (mean ± SEM) of FAS population day 61

(14) The observed diastolic blood pressure is shown in fig. 24.

Mean change from baseline (mean ± SEM) of FAS population day 61

(15) The observed hypersensitive C response protein (hsCRP) is shown in FIG. 25 and Table 15.

Mean change from baseline (LS mean.+ -. 95 CI) for day 61 of FAS population Table 15

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Claims

1. A method of reducing cardiovascular risk or mortality in a subject suffering from a symptom due to cardiovascular disease 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.

2. The method of claim 1, wherein the symptom is shortness of breath, shortness of breath on exercise, dizziness, chest pain, syncope, fatigue, impaired cardiac energy, or restricted activities of daily living.

3. The method of claim 2, wherein the daily living activity is limited to personal care, mobility, or eating difficulties.

4. The method of claim 1, wherein the cardiovascular disease comprises heart failure, a heart attack, coronary artery disease, or Coronary Heart Disease (CHD).

5. The method of claim 4, wherein the heart failure comprises heart failure with a preserved ejection fraction (HFpEF), heart failure with a reduced ejection fraction (hfrref), or heart failure with a moderate ejection fraction (HFmrEF).

6. The method of any one of claims 1-5, wherein the subject experiences a reduction in risk of a significant cardiovascular event following administration.

7. The method of claim 6, wherein the major cardiovascular event is death or hospitalization for exacerbation of the disease.

8. A method for treating HFpEF, HFrEF or HFmrEF in a subject, 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.

9. The method of claim 8, wherein the subject has at least one symptom selected from the group consisting of: shortness of breath, shortness of breath on exercise, impaired cardiac energy, dizziness, fatigue, dyspnea, palpitations (atrial tremor), chest discomfort, oedema, syncope and restricted activities of daily living.

10. The method of claim 9, wherein the activities of daily living are limited to personal care, mobility and eating difficulties.

11. The method of claim 8, wherein the subject has at least one symptom selected from the group consisting of: reduced exercise tolerance, fatigue, tiredness, increased recovery time after exercise, and ankle swelling.

12. The method of claim 8, wherein the subject has at least one symptom selected from the group consisting of: coronary artery disease, hypertension and heart murmurs.

13. The method of claim 1 or 8, wherein the subject experiences an improvement in cardiac bioenergy deficiency following administration, wherein the improvement comprises: a) weight loss >5%, b) blood pressure reduction, and/or c) a reduction in risk of a major cardiovascular event, wherein the major cardiovascular event is selected from the group consisting of death, hospitalization for exacerbation of the disease, and myocardial infarction.

14. The method of claim 1 or 8, further comprising administering a therapeutically effective amount of 5- [ (2, 4-dinitrophenoxy) methyl]Before and after-1-methyl-2-nitro-1H-imidazole, the peak oxygen consumption (VO) of the subject during exercise was assessed ₂ ) And/or VE/CO ₂ Or VE/VCO ₂ Slope, wherein VO in the subject after administration ₂ An increase in HFpEF, HFrEF, HFmrEF of the subject is indicative of a decrease in the extent of HFpEF, HFrEF, HFmrEF, or a decrease in one or more symptomatic components or conditions of the cardiovascular disease thereof.

15. The method of claim 1 or 8, wherein after administration, the method increases VO in the subject ₂ 。

16. The method of claim 1 or 8, further comprising assessing the 6 minute walking distance (6 MWD) of the subject during exercise before and after administration of a therapeutically effective amount of 5- [ (2, 4-dinitrophenoxy) methyl ] -1-methyl-2-nitro-1H-imidazole, wherein an increase in the 6MWD in the subject after administration is indicative of a reduction in the HFpEF level, or at least one symptomatic component or condition thereof, in the subject.

17. The method of claim 1 or 8, wherein the method increases the 6MWD after administration.

18. The method of claim 1 or 8, wherein the treatment further comprises evaluating the NYHA class score of the subject before and after administration.

19. The method of claim 18, wherein a decrease in NYHA score after administration is indicative of a decrease in the extent of heart disease in the subject.

20. The method of claim 19, wherein following administration, the method reduces the NYHA classification score of the subject from class III to class II, or from class II to class I.

21. A method of lowering blood pressure in a subject 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.

22. The method of claim 21, wherein the subject has cardiovascular disease, hypertension, refractory hypertension, or severe hypertension.

23. The method of claim 22, wherein the cardiovascular disease comprises heart failure, a heart attack, coronary artery disease, or Coronary Heart Disease (CHD).

24. The method of claim 23, wherein heart failure comprises HFpEF, HFrEF, or HFmrEF.

25. The method of any one of claims 21-24, wherein the subject has hypertension associated with HFpEF, HFrEF or HFmrEF.

26. The method of any one of claims 21-25, wherein the subject has at least one symptom selected from the group consisting of: headache, shortness of breath, chest pain, nose bleeding, dizziness, fatigue, vision problems, arrhythmia, hematuria, sweating, sleep difficulties and eye bloodstains.

27. The method of any one of claims 21 to 26, wherein reducing blood pressure comprises reducing diastolic pressure and/or reducing systolic pressure.

28. The method of claim 21, wherein the subject experiences a blood pressure decrease of at least 5mmHg after administration.

29. The method of claim 21, wherein the method reduces the risk of developing or slows progression of cardiovascular disease or heart failure.

30. The method of claim 29, wherein heart failure comprises HFpEF, HFrEF, or HFmrEF.

31. The method of any one of claims 1, 8 or 21, wherein the subject has obesity, hyperlipidemia, diabetes, hypertension, dyslipidemia, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, or metabolic syndrome.

32. A method of treating a cardiovascular disease comprising administering a therapeutically effective amount of 5- [ (2, 4-dinitrophenoxy) methyl ] -1-methyl-2-nitro-1H-imidazole or a pharmaceutically acceptable salt thereof in a subject so as to achieve at least one of:

v) AUC/C of about 18 _max Ratio.

33. The method of any one of claims 1 to 32, wherein after administration, the method does not create a clinically significant risk of adverse events.

34. The method of claim 33, wherein the adverse event comprises at least one of nausea, vomiting, sweating, dizziness, headache, cataracts, glaucoma, fever, hyperthermia, tachycardia, sweating, tachypnea, and death.

35. The method of claim 34, wherein the adverse event is characterized by at least one of elevated body temperature, increased heart rate, abnormal sweating, erythema, perspiration, dehydration, and abnormally rapid respiration.

36. A method of treating a subject's granline-related disorder or condition without a clinically significant risk of an adverse event, 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.

37. The method of claim 36, wherein the disorder is obesity, hyperlipidemia, 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 disease associated with increased Reactive Oxygen Species (ROS), friedreich's ataxia, or liver disease.

38. The method of claim 37, wherein the diabetes is type 2 diabetes (T2 DM).

39. The method of claim 37, wherein the cardiovascular disease comprises heart failure, HFpEF, HFrEF, HFmrEF, a heart attack, coronary artery disease, or CHD.

40. The method of claim 37, wherein the liver disease comprises non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), non-cirrhosis NASH with liver fibrosis, liver steatosis, liver fibrosis, liver cirrhosis, or hepatocellular carcinoma.

41. The method of claim 36, wherein the condition is at least one of steatosis, inflammation, fibrosis, cirrhosis, and hepatocyte damage in NASH.

42. The method of any one of claims 36-41, wherein after administration, the method provides at least one of the following in the subject:

v) AUC/C of about 18 _max Ratio.

43. The method of claim 36, wherein the adverse event is associated with a granosome uncoupling agent.

44. The method of claim 43 wherein the granulometric body uncoupler is 2, 4-dinitrophenol.

45. The method of any one of claims 36 to 44, wherein the adverse event comprises at least one of nausea, vomiting, sweating, dizziness, headache, cataracts, glaucoma, fever, hyperthermia, tachycardia, sweating, tachypnea, and death.

46. The method of claim 45, wherein the adverse event is characterized by at least one of elevated body temperature, increased heart rate, abnormal sweating, erythema, perspiration, dehydration, and abnormally rapid respiration.

47. The method of claim 45, wherein the adverse event is associated with cardiovascular collapse, cardiac arrest, and/or death.

48. The method of claim 47, wherein the adverse event is associated with cardiac arrest.

49. A method of reducing toxicity or side effects in treating a granline-related disorder or condition in a subject, 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.

50. The method of claim 49, wherein the disorder is obesity, hyperlipidemia, diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, cardiovascular disease, atherosclerosis, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, rattky's syndrome, metabolic syndrome associated with aging, metabolic disease associated with increased Reactive Oxygen Species (ROS), friedel-crafts ataxia, or liver disease.

51. The method of claim 50, wherein the diabetes is type 2 diabetes (T2 DM).

52. The method of claim 50, wherein the cardiovascular disease comprises heart failure, HFpEF, HFrEF, HFmrEF, a heart attack, coronary artery disease, or CHD.

53. The method of claim 50, wherein the liver disease comprises non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), non-cirrhosis NASH with liver fibrosis, liver steatosis, liver fibrosis, liver cirrhosis, or hepatocellular carcinoma.

54. The method of claim 49, wherein the condition is at least one of steatosis, inflammation, fibrosis, cirrhosis, and hepatocyte damage in NASH.

55. The method of claim 49, wherein the side effect is associated with a granosome decoupling agent.

56. The method of claim 55, wherein the granulometric body uncoupler is 2, 4-dinitrophenol.

57. The method of claim 56, comprising at least one of:

i) Extending half-life (t) of 2, 4-dinitrophenol _1/2 )；

iv) increasing the area under the curve (AUC).

58. The method of claim 57, wherein the average half-life is extended to about 20-50 hours, 25-40 hours, or 30-40 hours.

59. The method of claim 57, wherein the median T _max Extending to at least 6 hours or at least 8 hours.

60. The method of claim 57, wherein the median T _max Extending to about 6-8 hours or about 6-10 hours.

61. The method according to claim 57 wherein 2, 4-dinitrophenol C is reduced _max Comprising providing in the subject from about 80ng/mL to about 8300ng/mL of C of 2, 4-dinitrophenol after administration _max Is a steady state of (c).

62. The method of claim 57, wherein the method provides an AUC/C of about 18 in the subject _max Ratio.

63. The method of any one of claims 49-62, wherein the side effects comprise at least one of nausea, vomiting, sweating, dizziness, headache, cataracts, glaucoma, fever, hyperthermia, tachycardia, sweating, tachypnea, and death.

64. The method of claim 63, wherein the side effect is characterized by at least one of elevated body temperature, increased heart rate, abnormal sweating, erythema, perspiration, dehydration, and abnormally rapid respiration.

65. The method of claim 63, wherein the side effects are associated with cardiovascular collapse, cardiac arrest, and/or death.

66. The method of claim 65, wherein the side effect is associated with cardiac arrest.

67. A method of preventing overdose in treating a granline-related disorder or condition in a subject, 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.

68. The method of claim 67, wherein the disorder is obesity, hyperlipidemia, diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, cardiovascular disease, atherosclerosis, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, retter's syndrome, metabolic syndrome associated with aging, metabolic disease associated with increased Reactive Oxygen Species (ROS), friedreich's ataxia, or liver disease.

69. The method of claim 68, wherein the diabetes is type 2 diabetes (T2 DM).

70. The method of claim 68, wherein the cardiovascular disease comprises heart failure, HFpEF, HFrEF, HFmrEF, a heart attack, coronary artery disease, or CHD.

71. The method of claim 68, wherein the liver disease comprises non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), non-cirrhosis NASH with liver fibrosis, liver steatosis, liver fibrosis, liver cirrhosis, or hepatocellular carcinoma.

72. The method of claim 67, wherein the condition is at least one of steatosis, inflammation, fibrosis, cirrhosis, and hepatocyte damage in NASH.

73. The method of claim 67, wherein the overdose is associated with a granosome decoupling agent.

74. The method of claim 73 wherein the granulometric body uncoupler is 2, 4-dinitrophenol.

75. The method of claim 74, wherein following administration, the method provides in the subject at least one of:

v) AUC/C of about 18 _max Ratio.

76. The method of any one of claims 67 to 75, wherein said method does not create a clinically significant risk of adverse events.

77. The method of claim 76, wherein the adverse event comprises at least one of nausea, vomiting, sweating, dizziness, headache, cataracts, glaucoma, fever, hyperthermia, tachycardia, sweating, tachypnea, and death.

78. The method of claim 76, wherein the adverse event is characterized by at least one of elevated body temperature, increased heart rate, abnormal sweating, erythema, perspiration, dehydration, and abnormally rapid respiration.

79. The method of claim 76, wherein the adverse event is associated with cardiovascular collapse, cardiac arrest, and/or death.

80. The method of claim 79, wherein the adverse event is associated with cardiac arrest.

81. A method for increasing the metabolic rate of a subject without causing a clinically significant risk of an adverse event, 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.

82. The method of claim 81, wherein the subject has at least one of: obesity, hyperlipidemia, type 2 diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, atherosclerosis, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, rett's syndrome, metabolic syndrome associated with aging, metabolic diseases associated with increased Reactive Oxygen Species (ROS), friedreich's ataxia, NAFLD, NASH, non-cirrhosis NASH with liver fibrosis, liver steatosis, liver fibrosis, cirrhosis and hepatocellular carcinoma.

83. The method of claim 81 or 82, comprising increasing resting metabolic rate without causing clinically significant risk of adverse events.

84. The method of claim 83, wherein the resting metabolic rate is increased by at least 10%.

85. The method of claim 83, wherein the resting metabolic rate is increased by at least 20%.

86. A method for increasing resting energy expenditure in a subject, 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.

87. The method of claim 86, wherein the subject has at least one of: obesity, hyperlipidemia, type 2 diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, atherosclerosis, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, rett's syndrome, metabolic syndrome associated with aging, metabolic diseases associated with increased Reactive Oxygen Species (ROS), friedreich's ataxia, NAFLD, NASH, non-cirrhosis NASH with liver fibrosis, liver steatosis, liver fibrosis, cirrhosis and hepatocellular carcinoma.

88. The method of any one of claims 81-87, wherein the subject experiences an increase in resting energy expenditure of at least 10% after administration.

89. The method of any one of claims 81-87, wherein the subject experiences an increase in resting energy expenditure of at least 20% after administration.

90. The method of any one of claims 81-87, wherein the subject experiences an increase in resting energy expenditure of about 30% after administration.

91. A method for treating a metabolic disorder in a subject, 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.

92. The method of claim 91, wherein the subject has at least one of: obesity, hyperlipidemia, type 2 diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, atherosclerosis, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, rett's syndrome, metabolic syndrome associated with aging, metabolic diseases associated with increased Reactive Oxygen Species (ROS), friedreich's ataxia, NAFLD, NASH, non-cirrhosis NASH with liver fibrosis, liver steatosis, liver fibrosis, cirrhosis and hepatocellular carcinoma.

93. The method of claim 91, wherein the subject has at least one of: increased blood pressure, hyperglycemia, lumbar peri-body hyperlipidemia, and abnormal cholesterol or triglyceride levels.

94. The method of any one of claims 86-93, wherein after administration, the method does not create a clinically significant risk of adverse events in the subject.

95. The method of any one of claims 81-94, wherein the adverse event is associated with a granosome decoupling agent.

96. The method of claim 95 wherein the granulometric body uncoupler is 2, 4-dinitrophenol.

97. The method of claim 96, wherein following administration, the method provides in the subject at least one of:

v) AUC/C of about 18 _max Ratio.

98. The method of any one of claims 94-97 wherein the adverse event comprises at least one of nausea, vomiting, sweating, dizziness, headache, cataracts, glaucoma, fever, hyperthermia, tachycardia, sweating, tachypnea, and death.

99. The method of claim 98, wherein the adverse event is characterized by at least one of elevated body temperature, increased heart rate, abnormal sweating, erythema, perspiration, dehydration, and abnormally rapid respiration.

100. The method of claim 98, wherein the adverse event is associated with cardiovascular collapse, cardiac arrest, and/or death.

101. The method of claim 100, wherein the method results in cardiovascular adverse events in less than xx% of patients during xx day treatment periods.

102. The method of any of the preceding claims, further comprising administering a therapeutically effective amount of 5- [ (2, 4-dinitrophenoxy) methyl]-a step of determining in the subject, before and after 1-methyl-2-nitro-1H-imidazole, at least one of: steady state model assessment of insulin resistance (HOMA-IR); fasting blood glucose concentration; concentration of glycated albumin; glycosylated hemoglobin (hemoglobin A1c, hbA) _1c )。

103. The method of claim 102, wherein the subject experiences a decrease in at least one of body weight, blood pressure, and blood glucose following administration.

104. The method of claim 103, wherein the subject experiences at least one of:

i) Weight loss of at least 5% or at least 10%;

ii) a decrease in blood pressure of at least 5mmHg;

iii)HbA _1c a reduction of at least 0.5% or at least 1.5%;

iv) lipid reduction of at least 10%; and

v) liver fat reduction by at least 50%.

105. The method of any one of claims 81 to 104, wherein the method slows down the progress of at least one of: atherosclerosis, NAFLD, NASH, non-cirrhosis NASH with liver fibrosis, liver steatosis, liver fibrosis, liver cirrhosis and hepatocellular carcinoma.

106. The method of any one of claims 81-105, wherein the method accelerates a natural process in humans that improves a cardiovascular metabolic process.

107. A method of treating hypertriglyceridemia associated with cardiovascular disease, atherosclerosis, obesity, hypertension, diabetes, insulin resistance, and/or liver disease in a subject 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.

108. The method of claim 107, wherein the cardiovascular disease comprises heart failure, HFpEF, HFrEF, HFmrEF, a heart attack, coronary artery disease, or CHD.

109. The method of claim 107, wherein the liver disease comprises non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), non-cirrhosis NASH with liver fibrosis, liver steatosis, liver fibrosis, liver cirrhosis, or hepatocellular carcinoma.

110. The method of claim 107, wherein the subject has at least one of abdominal pain, pain in the mid-abdominal, thoracic or back regions, gastrointestinal pain, dyspnea, loss of appetite, nausea, vomiting, pancreatic inflammation, memory loss, dementia, macular tumors, corneal arcas, and xanthomas.

111. The method of any one of claims 107-110, wherein 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.

112. A method of treating severe hypertriglyceridemia in a subject 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.

113. The method of claim 112, wherein the subject has at least one of abdominal pain, pain in the mid-abdominal, thoracic or back regions, gastrointestinal pain, dyspnea, loss of appetite, nausea, vomiting, pancreatic inflammation, memory loss, dementia, macular tumors, corneal arcas, and xanthomas.

114. The method of claim 112 or 113, wherein the subject has a triglyceride blood level above 500 mg/dL.

115. The method of any one of claims 112-114, wherein the subject has refractory severe hypertriglyceridemia.

116. The method of any one of claims 112-115, wherein the subject has severe hypertriglyceridemia associated with cardiovascular disease, atherosclerosis, obesity, hypertension, diabetes, insulin resistance, and/or liver disease.

117. The method of any one of claims 112-116, wherein the subject is an adult male subject.

118. The method of any one of claims 112-117, wherein the subject is spanish progeny.

119. The method of any one of claims 107-118, wherein the method comprises reducing low density lipoprotein cholesterol levels and/or reducing non-high density lipoprotein cholesterol levels.

120. The method of claim 119, wherein the method comprises at least one of:

i) Lowering triglyceride levels by at least 5%, at least 10% or at least 20%;

121. The method of any one of claims 107-120, wherein after administration, the method slows progression of at least one of cardiovascular disease, atherosclerosis, obesity, hypertension, diabetes, insulin resistance, and/or liver disease; and/or reduce the risk of significant cardiovascular events.

122. The method of claim 121, wherein the significant cardiovascular event is death or hospitalization for exacerbation of the disease.

123. The method of any one of claims 107-122, wherein after administration, the method does not create a clinically significant risk of adverse events in the subject.

124. The method of claim 123, wherein the adverse event is associated with a granosome uncoupling agent.

125. The method of claim 124 wherein the granulometric decoupling agent is 2, 4-dinitrophenol.

126. The method of any one of claims 123-125, wherein the adverse event comprises at least one of nausea, vomiting, sweating, dizziness, headache, cataracts, glaucoma, fever, hyperthermia, tachycardia, sweating, tachypnea, and death.

127. The method of claim 126, wherein the adverse event is characterized by at least one of elevated body temperature, increased heart rate, abnormal sweating, erythema, perspiration, dehydration, and abnormally rapid respiration.

128. The method of claim 126, wherein the adverse event is associated with cardiovascular collapse, cardiac arrest, and/or death.

129. The method of claim 128, wherein the adverse event is a cardiac arrest.

130. A method of reducing liver fat in a subject by at least 50% 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.

131. The method of claim 130, wherein the subject has non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), and/or liver steatosis.

132. The method of claim 130 or 131, further comprising the step of: determining the subject before and after administrationVibration Controlled Transient Elastography (VCTE),/>Controlled Attenuation Parameter (CAP) score, magnetic resonance imaging proton density fat fraction (MRI-PDFF), and Enhanced Liver Fibrosis (ELF) score.

133. The method of claim 132, wherein prior to administration, the subject has a CAP score of greater than 300 dB/m.

134. The method of claim 132, wherein prior to administration, the subject has at least 8% liver fat by MRI-PDFF.

135. A method of reducing lipid in a subject by at least 10% 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.

136. The method of claim 135, wherein the subject has at least one of: obesity, hyperlipidemia, type 2 diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, atherosclerosis, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, rett's syndrome, metabolic syndrome associated with aging, metabolic diseases associated with increased Reactive Oxygen Species (ROS), friedreich's ataxia, NAFLD, NASH, non-cirrhosis NASH with liver fibrosis, liver steatosis, liver fibrosis, cirrhosis and hepatocellular carcinoma.

137. The method of claim 135 or 136, further comprising the step of: before and after administration, the subject was determined for serum hypersensitive 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).

138. The method of any one of claims 129 to 137, wherein the method provides at least one of:

i) Weight loss of at least 10%;

ii)HbA _1c a reduction of at least 0.5% or at least 1.5%;

iii) A decrease in blood pressure of at least 5mmHg;

iv) lipid reduction of at least 10%;

v) liver fat reduction of at least 50%;

vi) reduction of serum alanine Aminotransferase (ALT); and

vii) reduction of aspartate Aminotransferase (AST).

139. The method of any one of claims 129-138, wherein after administration, the method does not create a clinically significant risk of an adverse event in the subject.

140. The method of claim 139, wherein the adverse event is associated with a granosome uncoupling agent.

141. The method of claim 140 wherein the granulometric body uncoupler is 2, 4-dinitrophenol.

142. The method of any one of claims 138-140, wherein the adverse event comprises at least one of nausea, vomiting, sweating, dizziness, headache, cataracts, glaucoma, fever, hyperthermia, tachycardia, sweating, tachypnea, and death.

143. The method of claim 142, wherein the adverse event is characterized by at least one of elevated body temperature, increased heart rate, abnormal sweating, erythema, perspiration, dehydration, and abnormally rapid respiration.

144. The method of claim 142, wherein the adverse event is associated with cardiovascular collapse, cardiac arrest, and/or death.

145. The method of claim 144, wherein the adverse event is associated with cardiac arrest.

146. The method of any one of claims 107-145 wherein after administration of about 30mg to about 1400mg of 5- [ (2, 4-dinitrophenoxy) methyl ] -1-methyl-2-nitro-1H-imidazole, the method provides in the subject at least one of:

v) AUC/C of about 18 _max Ratio.

147. A method of treating or reducing the risk of cancer in a subject 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.

148. The method of claim 147, wherein the cancer comprises biliary tract cancer, bladder cancer, brain cancer (i.e., meningioma), breast cancer (post-menopausal), cervical cancer, colorectal cancer, endometrial/uterine cancer, esophageal cancer, gall bladder cancer, head and neck cancer, kidney cancer, leukemia, liver cancer, multiple myeloma, non-hodgkin's lymphoma, ovarian cancer, pancreatic cancer, gastric and thyroid cancer, and prostate cancer.

149. The method of claim 147 or 148, wherein the cancer is associated with obesity, hyperlipidemia, diabetes, hypertension, dyslipidemia, metabolic disease, liver disease, and/or cardiovascular disease.

150. A method of treating obesity, excessive body fat, type 2 diabetes, insulin resistance or intolerance, hypertension, dyslipidemia, atherosclerosis, hypertriglyceridemia, acquired lipodystrophy, hereditary lipodystrophy, partial lipodystrophy, metabolic syndrome, rett's syndrome, metabolic syndrome associated with aging, metabolic diseases associated with increased Reactive Oxygen Species (ROS), friedreich ataxia, NAFLD, NASH, non-cirrhosis NASH with liver fibrosis, liver steatosis, liver fibrosis, liver cirrhosis or hepatocellular carcinoma, the 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, in a subject so as to achieve at least one of:

v) AUC/C of about 18 _max Ratio.

151. The method of any one of the preceding claims, wherein the subject has an elevated Body Mass Index (BMI).

152. The method of claim 151, wherein the subject has about 28.0kg/m ² To about 45.0kg/m ² Is a BMI of (B).

153. The method of any one of the preceding claims, wherein the subject to be treated meets inclusion and exclusion criteria of example 5 or example 6.

154. The method of any one of the preceding claims, wherein prior to administration, the subject is in a fasting condition.

155. The method of any one of the preceding claims, wherein prior to administration, the subject is in fed condition.

156. The method of any one of the preceding claims, wherein the therapeutically effective amount is about 30mg to about 1400mg per day, about 100mg to about 1000mg per day, about 150mg to about 600mg per day, or 200mg to 550mg per day.

157. The method of any one of the preceding claims, wherein the therapeutically effective amount 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 per day.

158. The method of claim 157, wherein the therapeutically effective amount is about 30mg, 100mg, 200mg, 500mg, 600mg, 1050mg, or 1400mg per day.

159. The method of claim 157, wherein the therapeutically effective amount is about 200mg, 400mg, or 550mg per day.

160. The method of claim 157, wherein the therapeutically effective amount is about 170mg, 340mg, 510mg per day.

161. The method of claim 157, wherein the therapeutically effective amount is about 150mg, 300mg, 450mg per day.

162. The method of any of the preceding claims wherein 5- [ (2, 4-dinitrophenoxy) methyl ] -1-methyl-2-nitro-1H-imidazole is administered orally once daily.

163. The method of any of the preceding claims, wherein 5- [ (2, 4-dinitrophenoxy) methyl ] -1-methyl-2-nitro-1H-imidazole is administered once daily for two weeks, four weeks, six weeks, eight weeks, ten weeks, one month, two months, three months, four months, six months, eight months, or one year.

164. The method of any of the preceding claims wherein 5- [ (2, 4-dinitrophenoxy) methyl ] -1-methyl-2-nitro-1H-imidazole is administered in hydroxypropyl methylcellulose capsules.