High Throughput Assay for Screening Compounds Against T. Brucei

Sponsor: NIH National Institute of Neurological Disorders and Stroke

Location(s): United States


Parasitic diseases represent major global health problems affecting hundreds of millions of people. Because these are diseases primarily of developing countries there is little interest in drug development by the pharmaceutical industry. Drug discovery and development must therefore be an extremely cost-effective process. The Sandier Center for Research in Parasitic Diseases in conjunction with the Bay Area Screening Center is poised to establish high throughput screening assays suitable for evaluating moderate to large compound libraries as sources for new anti-parasitic drug leads. As a first step in this effort, we have developed a "cell-based" screening assay for Trypanosoma brucei, the causative agent of African sleeping sickness. This microtiter-based assay, which uses the luciferase reaction to detect ATP regeneration by the parasite, has been shown to accurately identify trypanocidal drugs in a preliminary study. We are seeking support to transfer this assay to our High Throughput Screening Facility, optimizing robotic liquid handling for each step of the assay, and validating its utility by pilot screening against a subset of the Chembridge and Chemdiv libraries already in-house. When the pilot study is complete and optimized, we will screen 2160 compounds of primarily FDA-approved drugs. We will miniaturize the current assay to 384 well plates for the goal of screening up to 10,000 compounds/day. Results with our luciferase assay will be cross validated with the Alamar blue assay and counter screened for mammalian cytotoxicity. This study will serve as a model for parallel development of similar cell-based assays for other parasite infections of global health significance. We have established the in-house expertise, equipment and synthetic chemistry capability to allow us, in the future, to exploit results from this initial study to rationally choose specific leads to expand in secondary diverse compound libraries. Computational and informatics capability is in place to acquire, store and cross-reference this data, and provide access to the scientific community through an open-source website.