Metabolic syndrome (MetS), a constellation of medical conditions that includes obesity, type 2 diabetes, fattyliver, hypertension, cardiovascular disease, and dyslipidemia, creates a major burden for individuals andsociety. MetS is caused by insulin resistance and obesity, and a drug that could reverse these two conditionswould transform its treatment. Unfortunately, no safe and effective medication with such properties exists.Equator Therapeutics is developing a first-in-class drug that will increase the resting metabolic rate byactivating the native pathways for mitochondrial H+ leak and thermogenesis. This drug will correct the rootcause of metabolic disorders-the imbalance between excess caloric intake and limited energy expenditure-and is expected to be highly effective at reversing obesity and insulin resistance. Until recently, development ofsuch drug was impossible due to a very limited understanding of the mechanisms of the mitochondrial H+ leak.Instead, protonophores that indiscriminately increase H+ leak not only in mitochondria but all cell membranes,were used, leading to undesirable side effects. In contrast, our drug will activate H+ leak specifically inmitochondria via the physiologically regulated native H+ leak pathways mediated by uncoupling proteins and isexpected to be much safer. The development of this drug is enabled by our recent discoveries concerning themolecular mechanisms of the mitochondrial H+ leak. In Specific Aim 1, we will identify a set of diversedrug-like compounds that selectively activate a protein target natively responsible for themitochondrial H+ leak. Importantly, these compounds will have no protonophoric activity. We have selected alibrary of ~67,000 small molecules with desirable chemical properties specific to our target. We will screen thislibrary for hits that increase mitochondrial H+ leak using new high-throughput mitochondrial assays consistingof a primary screen and counter-screen to remove false positives. Compounds that pass both rounds of high-throughput screening will be validated by direct measurement of the H+ current via our target protein usingmitochondrial patch-clamp electrophysiology. Additional patch-clamp studies in the plasma membrane willfurther select only those of them that lack any protonophoric activity. In Specific Aim 2, we will determine theeffects of the novel activators of the native mitochondrial H+ leak pathways on the bioenergetics ofintact human cells in vitro. We will primarily focus on the ability of our compounds to stimulate uncoupledrespiration, the parameter reflecting their ability to activate mitochondrial H+ leak and thermogenesis in intactcells. Other parameters such as the respiration capacity of the cell, activity of the glycolytic pathways, and theintracellular ATP level will also be assessed. Finally, cytotoxicity assays will be performed. Successfulaccomplishment of these aims will generate a set of chemically diverse lead compounds that we will bringforward to animal studies with the goal of developing a safe drug to transform treatment of MetS.
Public Health Relevance Statement: Project Narrative This project will identify molecules that activate native pathways for mitochondrial thermogenesis, with the goal of developing a drug to safely increase metabolic rate. This drug will reverse insulin resistance and obesity and will have potential to transform the treatment of metabolic disorders. 1
Project Terms: Adenine Nucleotides ; Adenosine Phosphates ; Adult ; 21+ years old ; Adult Human ; adulthood ; Animals ; inhibitor/antagonist ; inhibitor ; Automobile Driving ; driving ; Bioenergetics ; Biological Assay ; Assay ; Bioassay ; Biologic Assays ; Brown Fat ; Brown Adipose Tissue ; Hibernating Gland ; Energy Intake ; Caloric Intake ; caloric dietary content ; Cardiovascular Diseases ; cardiovascular disorder ; Cell Line ; CellLine ; Strains Cell Lines ; cultured cell line ; Cell membrane ; Cytoplasmic Membrane ; Plasma Membrane ; plasmalemma ; Cells ; Cell Body ; Citric Acid Cycle ; Krebs Cycle ; TCA cycle ; Tricarboxylic Acid Cycle ; Diabetes Mellitus ; diabetes ; Non-Insulin-Dependent Diabetes Mellitus ; Adult-Onset Diabetes Mellitus ; Ketosis-Resistant Diabetes Mellitus ; Maturity-Onset Diabetes Mellitus ; NIDDM ; Non-Insulin Dependent Diabetes ; Noninsulin Dependent Diabetes ; Noninsulin Dependent Diabetes Mellitus ; Slow-Onset Diabetes Mellitus ; Stable Diabetes Mellitus ; T2 DM ; T2D ; T2DM ; Type 2 Diabetes Mellitus ; Type 2 diabetes ; Type II Diabetes Mellitus ; Type II diabetes ; adult onset diabetes ; ketosis resistant diabetes ; maturity onset diabetes ; type 2 DM ; type II DM ; type two diabetes ; Dinitrophenols ; Pharmaceutical Preparations ; Drugs ; Medication ; Pharmaceutic Preparations ; drug/agent ; Electron Transport ; electron transfer ; Electrophysiology (science) ; Electrophysiology ; Neurophysiology / Electrophysiology ; electrophysiological ; Energy Metabolism ; Energy Expenditure ; Epidemic ; Fatty Liver ; Liver Steatosis ; hepatic steatosis ; Glycolysis ; Goals ; Thermogenesis ; Heat Production ; Human ; Modern Man ; Hypertension ; Vascular Hypertensive Disease ; Vascular Hypertensive Disorder ; high blood pressure ; hyperpiesia ; hyperpiesis ; hypertensive disease ; In Vitro ; Insulin Resistance ; insulin resistant ; Lead ; Pb element ; heavy metal Pb ; heavy metal lead ; Libraries ; Liver diseases ; Hepatic Disorder ; hepatic disease ; hepatopathy ; liver disorder ; Metabolic Diseases ; Metabolic Disorder ; Thesaurismosis ; metabolism disorder ; Mitochondria ; mitochondrial ; Obesity ; adiposity ; corpulence ; Proteins ; Respiration ; respiratory mechanism ; Risk ; Running ; Safety ; Societies ; Specificity ; Testing ; Tissues ; Body Tissues ; Weight Gain ; Weight Increase ; body weight gain ; body weight increase ; wt gain ; Body Weight decreased ; Weight Loss ; Weight Reduction ; body weight loss ; wt-loss ; 2,4-Dinitrophenol ; 2,4-DNP ; DNP ; Generations ; Measures ; Mediating ; uncoupling protein 1 ; BAT uncoupling protein ; brown adipose tissue uncoupling protein ; thermogenin ; PPAR gamma ; PPAR-γ ; PPARgamma ; PPARγ ; Peroxisome Proliferative Activated Receptor Gamma ; Peroxisome Proliferator-Activated Receptor gamma ; Peroxisome Proliferator-Activated Receptor γ ; Thiazolidinedione Receptor ; improved ; Area ; Clinical ; Physiological ; Physiologic ; Medical ; Series ; Chemicals ; Hepatocyte ; Hepatic Cells ; Hepatic Parenchymal Cell ; Liver Cells ; Inner mitochondrial membrane ; insight ; Individual ; Dyslipidemias ; Measurement ; Patch-Clamp Techniques ; Patch-Clamp Technics ; Plant Roots ; root ; chemical property ; Agonist ; Therapeutic ; Nature ; extracellular ; Lytotoxicity ; cytotoxicity ; professor ; Basal Metabolism ; resting metabolic rate ; Basal metabolic rate ; novel ; Position ; Positioning Attribute ; Regulation ; Modeling ; Property ; drug development ; High Throughput Assay ; high throughput screening ; patch clamp ; Metabolic syndrome ; Seahorse ; Genus Hippocampus ; ADP Translocase ; ADP,ATP Carrier ; ADP,ATP Translocator Protein ; ATP Translocase ; ATP-ADP Translocase ; Adenine Nucleotide Translocase ; small molecule ; Address ; Molecular ; Knock-out ; Knockout ; Development ; developmental ; Pathway interactions ; pathway ; insulin sensitizing drugs ; insulin sensitizer ; mouse model ; murine model ; ratiometric ; counterscreen ; metabolic rate ; side effect ;
