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Identification of Potent Inhibitors of the Trypanosoma bruceiMethionyl-tRNA Synthetase via High-Throughput Orthogonal Screening

Authors :
PedrĂ³-Rosa, Laura
Buckner, Frederick S.
Ranade, Ranae M.
Eberhart, Christina
Madoux, Franck
Gillespie, J. Robert
Koh, Cho Yeow
Brown, Steven
Lohse, Jacqueline
Verlinde, Christophe L.M.
Fan, Erkang
Bannister, Thomas
Scampavia, Louis
Hol, Wim G.J.
Spicer, Timothy
Hodder, Peter
Source :
SLAS Discovery: Advancing Life Sciences R&D; January 2015, Vol. 20 Issue: 1 p122-130, 9p
Publication Year :
2015

Abstract

Improved therapies for the treatment of Trypanosoma brucei, the etiological agent of the neglected tropical disease human African trypanosomiasis, are urgently needed. We targeted T. bruceimethionyl-tRNA synthetase (MetRS), an aminoacyl-tRNA synthase (aaRS), which is considered an important drug target due to its role in protein synthesis, cell survival, and its significant differences in structure from its mammalian ortholog. Previous work using RNA interference of MetRS demonstrated growth inhibition of T. brucei, further validating it as an attractive target. We report the development and implementation of two orthogonal high-throughput screening assays to identify inhibitors of T. bruceiMetRS. First, a chemiluminescence assay was implemented in a 1536-well plate format and used to monitor adenosine triphosphate depletion during the aminoacylation reaction. Hit confirmation then used a counterscreen in which adenosine monophosphate production was assessed using fluorescence polarization technology. In addition, a miniaturized cell viability assay was used to triage cytotoxic compounds. Finally, lower throughput assays involving whole parasite growth inhibition of both human and parasite MetRS were used to analyze compound selectivity and efficacy. The outcome of this high-throughput screening campaign has led to the discovery of 19 potent and selective T. bruceiMetRS inhibitors.

Details

Language :
English
ISSN :
24725552 and 24725560
Volume :
20
Issue :
1
Database :
Supplemental Index
Journal :
SLAS Discovery: Advancing Life Sciences R&D
Publication Type :
Periodical
Accession number :
ejs59342161
Full Text :
https://doi.org/10.1177/1087057114548832