Summary NAD+ is biosynthesized mainly through the salvage pathway and is central for cellular homeostasis and energy production. Deficiency of NAD+ results in disruption of cellular functions, leading to decreased ability to regenerate and repair. In heart failure (HF), the heart undergoes metabolic disruption leading to redox imbalance with increased NADH/NAD+ ratio. Therefore, we hypothesized that “Activating NAD+ plays a central role in attenuating Heart failure.” Current therapeutics manage the disease symptoms, however, do not address the core issues of HF. Based on our innovation with US patents and new preliminary data, Nampt activator P7C3 is a prototype for generation of new pharmacological class of small molecules for cardioprotection. Using medicinal chemistry, 3D-SAR, cell permeability and drug design approaches we will activate NAD+ by P7C3-based novel small molecules for attenuating HF. A combination and tiered approach will be utilized for evaluating and narrowing down the new compound pipeline using in vitro models, followed with efficacy studies. Initial screening through in vitro assay will identify the molecules that are of high affinity and activity. Subsequently, 8-9 new small molecules will be tested using ex vivo Langendorff system for evaluating the cardiac changes, and NAD+ generation capacity. Finally, 1-2 lead small molecule(s) will be tested using in vivo system with preclinical model of HF using isoproterenol infusion, alternate HF models include db/db and MI to evaluate the pharmacological specificity by using a combination of cardiac-specific Nampt KO and wildtype (C57Bl/6J) mice for demonstrating the increased function with NAD+ activation in HF. The milestone-based approach for R61 phase will be utilized for medicinal chemistry, including design and synthesis of new small molecules, in vitro Nampt activity, cell permeability and toxicity, and assessment of therapeutic candidate's ex vivo. Whereas, in the R33 phase, we will perform pre-clinical assessment and pharmacological evaluation of 1-2 lead small molecule(s) in vivo. The proposal provides a systematic plan for continuous assessment and refinement of project management to develop P7C3-based small molecules for testing and refinement to achieve the milestones in a timely manner. The University of South Florida is a top producer of patents and is highly supportive with cost matching during R33 phase. In addition, the USF hosts incubator companies which along with pharmaceutical research companies in Tampa Bay area have shown interest and support for this project. The PI is an expert and well known in the region with pharmaceutical incubator companies and has received Florida High Tech Corridor matching awards and therefore anticipate developing the lead compounds for commercialization through the next stage of development. Overall, the successful completion of the project will allow new product development with rigorous and robust evaluation along with project management of novel small molecules for therapeutic development and discovery of innovative new class of “NAD+ activators” for translational development.

PROJECT NARRATIVE The proposed studies will identify novel small molecules using patented prototype P7C3 based discovery pipeline for Heart failure treatment. The new P7C3 based novel small molecule discovery and synthesis will allow to optimize the efficacy, cell permeability and EC50 for higher therapeutic potential. Overall, the project will generate high impact translational data with discovery of P7C3 (Nampt activator) based small molecules for attenuating cardiovascular diseases and new product development.