Validating choline metabolism as a drug target in the fight against T.brucei (360G-Wellcome-086658_Z_08_Z)

In this proposal we will use multi-disciplinary approaches to investigate choline phospholipid biosynthesis and metabolism in Trypanosoma brucei as a source of potential chemotherapeutic targets. Emphasis will be on understanding choline metabolic pathways, so that we can genetically and chemically validate them as drug targets. This project aims to answer three key questions; 1) T.brucei are auxotrophic for choline, so, how do they obtain choline? 2) Are T.brucei vulnerable to inhibit ion of de novo phosphatidylcholine synthesis? 3) Is catabolism of choline containing species essential to the parasite? Thus our key goals are to: genetically validate the choline branch of the Kennedy pathway (phosphatidylcholine de novo synthesis) as a drug target. understand the uptake and usage of choline containing species by bloodstream T.brucei. develop high-throughput assays for enzymes involved in choline metabolism. obtain kinetic, specificity and structu ral information about the enzymes involved in choline metabolism, that will allow the construction of structure-activity relationships to facilitate the design of more potent parasite-specific inhibitors. ultimately allowing development of potent inhibitors with activity against live trypanosomes compliant with Lipinski s rules, thus suitable for future development as novel anti-trypanosomal drugs.