Chemical genetic validation of drug targets for Chagas disease

Abstract

The validation of a molecular target is a fundamental step in the drug research and development process. This project aims to advance the characterization and validation of four "potential" targets of the protozoan Trypanosoma cruzi, which causes Chagas disease. These targets are the enzymes L-threonine dehydrogenase (TcTDH), inosine-5-monophosphate dehydrogenase (TcIMPDH), NADPH-dependent Malic Enzyme (TcME) and glucose-6-phosphate dehydrogenase (TcG6PDH). The TcTDH and TcIMPDH enzymes were partially characterized in the previous project (FAPESP Regular Aid, n. 2018/22202-8). In this project, we intend to complete the characterization of the allosteric regulation of these enzymes and expand the high-throughput screening experiments to select more potent and selective inhibitors for in vitro assays against the parasite. Concomitant to the studies with TcTDH and TcIMPDH, we will continue the chemical-genetic validation of the enzymes TcME and TcG6PDH, for which resistant variants were already established towards specific inhibitors of each target (preliminary results). These inhibitors were identified by HTS trials performed previously (Mercaldi, G.F., 2014 and Ranzani, A. T, 2017) and the resistant variants were selected by the PhD student Amanda G. Eufrásio. If successful, this project will complete the requirements to move drug candidates, with defined mode of action, to the pre-clinical stages of the development of new drugs against Chagas disease. (AU)