Exploring trypanosoma brucei brucei protein targets for drug discovery.

• Main Author: Cheong, Yue Sheng.
• Other Authors: School of Biological Sciences
• Format: Final Year Project
• Language: English
• Published: 2013
• Subjects: DRNTU::Science
• Online Access: http://hdl.handle.net/10356/52455
• Institution: Nanyang Technological University

We created novel recombinant plasmids harbouring the reporter genes green fluorescent protein (GFP), DsRed and Renilla luciferase respectively, and tagged DsRed to partial length invariant surface glycoprotein 65 (ISG65) by overlap PCR. Electroporation of GFP plasmid into bloodstream form Trypanosoma brucei brucei was performed, and found to be successful, albeit with a weak GFP signal. We have shown the feasibility of the parasite harbouring foreign reporter genes, and hope to introduce other reporter genes, including plasmids harbouring DsRed and Renilla luciferase which we have created, in future. We hope these novel strains will help pave the way for alternative live-cell assays for future experiments. We also tested a group of focused inhibitors targeting the cysteine proteases of the parasite, and shortlisted three Novartis proprietary compounds (NPCs), NPC 1, 3, and 4 that were more potent than L-3-trans-(propylcarbamoyl)oxirane-2-carbonyl]-L-isoleucyl-L-proline methyl ester (CA-074ME), our reference compound which also targets cysteine proteases. Moving forward, further chemistry derivatization will be carried out for these promising compounds to synthesize novel drugs to treat human African trypanosomiasis.