Nephrilin peptide is a novel designed inhibitor of Rictor complex (also known as mTORC2), an evolutionarily conserved assembly believed to coordinate cellular stress signals, among other functions. As demonstrated in twelve separate rodent studies reported in six published research papers as well as in the data section of this proposal, inhibition of Rictor complex by daily subcutaneous bolus administration of nephrilin reverses the effects of stress in numerous animal models. Specifically, in burned animals, nephrilin administered beginning two hours after burn injury and continuing for seven days mitigates the loss of glycemic control, kidney function and lean body mass, and reverses increases in inflammation and sepsis mortality. One recent study has documented important effects of nephrilin treatment on the rat central nervous system after burn trauma. Nephrilin treatment following burn injury reverses epigenetic and signaling changes in kidney tissue leading to activation of Rac1, a key component of NADPH oxidase, as well as elevations in markers of oxidative stress such as urinary 8-isoprostane. Some, but not all, of the above effects of nephrilin appear to be influenced by the animal's iron status. Altered iron status has been observed in post-trauma human populations. Twenty-three grams of nephrilin were manufactured under GMP, quality assay methods were developed and validated and the manufactured drug product showed excellent stability over 18 months of storage. A CMC Section document was written. Although several wound healing products have been approved for use in burn patients, there is currently no pharmacologic intervention to treat the numerous severe and enduring systemic effects of burn trauma. Nephrilin is being developed for treatment of burn trauma with a main focus on these important systemic effects (not wound healing, for which products already exist). In Phase I of this SBIR project we will investigate the effect of iron supplementation and nephrilin dosage on the efficacy of nephrilin in a rat scald model. We will also complete the development of a method for bioassay of nephrilin in biological fluids and a product release bioassay for nephrilin potency. In Phase II, we will conduct GLP toxicology studies, transfer and validate methods and aseptic fill-and-finish of bulk drug substance under cGMP for a future clinical Phase I study, and develop a preliminary protocol for a future Phase 1 clinical trial.
Public Health Relevance Statement: Project Narrative Severe burn injury often causes an array of secondary complications, including sepsis, metabolic and organ dysfunction and cognitive changes, many of which can last a lifetime. At present most treatments are palliative and nephrilin is the first new drug to address the underlying epigenetic and signaling disruptions that lead to this vast array of secondary complications. Development of nephrilin will reduce long-term care costs and improve quality of life for victims of serious burns.
Project Terms: Address; Animal Model; Animals; base; Binding; Biological; Biological Assay; Bolus Infusion; Burn injury; Burn Trauma; care costs; Cellular Stress; Clinical; Clinical Research; cognitive change; Complex; Cyclic GMP; Data; design; Development; dosage; Dose; Drug Kinetics; Economics; Enzyme-Linked Immunosorbent Assay; Epigenetic Process; exposed human population; Fibroblasts; Functional disorder; Future; glycemic control; Goals; Health Care Costs; Hour; Human; Hydrogen Peroxide; improved; Inflammation; inhibitor/antagonist; Injections; Intervention; Iron; iron supplementation; Isoprostanes; Kidney; Lead; lean body mass; Liquid substance; Long-Term Care; Measures; Metabolic; Methods; Modeling; mortality; NADPH Oxidase; Neuraxis; novel; novel therapeutics; Organ; Oxidative Stress; Palliative Care; Paper; Patients; Peptides; Pharmaceutical Preparations; Pharmacology; Phase; phase 1 study; Phase I Clinical Trials; Population; preclinical development; Production; protective effect; Protocols documentation; Publishing; Quality of life; Rattus; Reactive Oxygen Species; Renal function; Reporting; Research; Rodent; Rodent Model; Sepsis; Serum; Serum iron level result; Signal Transduction; Small Business Innovation Research Grant; Solubility; Stress; Study models; subcutaneous; Tissues; Toxicokinetics; Toxicology; Trauma; United States; urinary; Wound Healing
