title: Yeast-based automated high-throughput screens to identify anti-parasitic lead compounds.
abstract: We have developed a robust, fully automated anti-parasitic drug-screening method that selects compounds specifically targeting parasite enzymes and not their host counterparts, thus allowing the early elimination of compounds with potential side effects. Our yeast system permits multiple parasite targets to be assayed in parallel owing to the strains' expression of different fluorescent proteins. A strain expressing the human target is included in the multiplexed screen to exclude compounds that do not discriminate between host and parasite enzymes. This form of assay has the advantages of using known targets and not requiring the in vitro culture of parasites. We performed automated screens for inhibitors of parasite dihydrofolate reductases, N-myristoyltransferases and phosphoglycerate kinases, finding specific inhibitors of parasite targets. We found that our 'hits' have significant structural similarities to compounds with in vitro anti-parasitic activity, validating our screens and suggesting targets for hits identified in parasite-based assays. Finally, we demonstrate a 60 per cent success rate for our hit compounds in killing or severely inhibiting the growth of Trypanosoma brucei, the causative agent of African sleeping sickness.
authors: Cambridge Systems Biology Centre and Department of Biochemistry, University of Cambridge, Sanger Building, 80 Tennis Court Road, Cambridge CB2 1GA, UK.
authors: Elizabeth
authors: E
authors: Bilsland
authors: Andrew
authors: A
authors: Sparkes
authors: Kevin
authors: K
authors: Williams
authors: Harry J
authors: HJ
authors: Moss
authors: Michaela
authors: M
authors: de Clare
authors: Pinar
authors: P
authors: Pir
authors: Jem
authors: J
authors: Rowland
authors: Wayne
authors: W
authors: Aubrey
authors: Ron
authors: R
authors: Pateman
authors: Mike
authors: M
authors: Young
authors: Mark
authors: M
authors: Carrington
authors: Ross D
authors: RD
authors: King
authors: Stephen G
authors: SG
authors: Oliver
mesh_terms: D000977:Antiparasitic Agents / Q000737:chemistry / Q000494:pharmacology*; D055808:Drug Discovery; D057166:High-Throughput Screening Assays; D006801:Humans; D007854:Lead / Q000737:chemistry* / Q000494:pharmacology; D054852:Small Molecule Libraries / Q000737:chemistry*; D014346:Trypanosoma brucei brucei / Q000187:drug effects; D014353:Trypanosomiasis, African / Q000188:drug therapy* / Q000473:pathology; D015003:Yeasts / Q000187:drug effects
chemical_list: D000977:Antiparasitic Agents; D054852:Small Molecule Libraries; D007854:Lead