Abstract 178: Development of a New Structural Family of Microbial Choline Trimethylamine Lyase Inhibitors for the Treatment and Prevention of Cardiovascular Disease

Abstract

Ashraf Duzan, BPharm., MS 1,2 ; Xiaodong Gu, Ph.D. 1,3 ; Adam Roberts, Ph.D. 1,3 ; Jennifier Buffa, MS 1,3 ; James Anderson 1,3 , Valentine Gogonea, Ph.D 1,2,3 Stanley L. Hazen, MD, Ph.D. 1,2,3 . 1 Department of Cellular & Molecular Medicine, 3 Center for Microbiome and Human Health, 4 Department of Cardiovascular Medicine, Cleveland Clinic, Cleveland, Ohio 44195; 2 Department of Chemistry, Cleveland State University, Cleveland, Ohio 44115 Recent clinical research points to trimethylamine N-oxide (TMAO), a gut microbiota generated metabolite, as a biomarker associated with acute thrombotic event and cardiovascular disease risks, and a direct causative contributor to these adverse phenotypes. Our goal in this project is to develop novel microbial enzyme inhibitors that alter the biosynthetic pathway of TMAO in vivo through the selective potent inhibition of gut microbial choline trimethylamine (TMA) lyase activity, the rate limiting step in TMA and TMAO generation in vivo. TMA generation by gut microbiota is predominantly catalyzed by the gut microbial enzyme pair CutC/D, members of the microbial choline utilization ( cut ) gene cluster. We are using structure activity relationships (SAR) to predict new classes of chemical structures as potential efficient inhibitors. Thus, we are exploring the synthesis of new chemical compounds, non-lethal to the gut microbial community, with high inhibitory potency in multiple in vitro assays employing evolutionarily diverse microbial cutC/D (primary and secondary screens), polymicrobial communities (tertiary screens), and for inhibitors that pass the above screening assays, progression to in vivo studies. We are preparing and assessing inhibitors that can work either through irreversible non-competitive or competitive mechanisms, possess appropriate physico-chemical pharmaceutical properties and have minimal systemic exposure to the host in effort to minimize possibility of side effects. Our leading candidates have excellent enzyme blocking efficiency and display good pharmacokinetic/pharmacodynamics properties.