Screening For Inhibitors Of Essential Leishmania Glucose Transporters
Annual rept. 1 Jul 2010-30 Jun 2011
OREGON HEALTH AND SCIENCE UNIV PORTLAND
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The major objective of this project is to identify compounds that function as selective inhibitors of the essential glucose transporters of the parasite Leishmania mexicana. Toward this end, a cell growth assay has been developed that can be employed in a high throughput screen HTS for such inhibitors. The dye SYBR green has been identified as the optimal reagent for following cell growth by fluorescence. The assay has been optimized initially for a 96-well plate assay and subsequently for a higher density 384-well plate assay. Statistical criteria have been employed to demonstrate that the assay functions with high accuracy and reproducibility under control conditions, with a Z -factor of approximately 0.9. This assay was subsequently employed in a scaling screen of the 2000 compound MicroSource Discovery Spectrum Collection to yield a Z-factor of 0.84 in the screen of a real library. The high quality of the assay indicates that it is ready to proceed to an HTS of a much larger library. The assay has been transferred to the HTS facility at St. Jude Children s Research Hospital in Memphis, TN to initiate full high throughput screens.
- Medicine and Medical Research