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ACS Infect Dis. 2016 Apr 8;2(4):281-293. Epub 2016 Feb 10.

High-Throughput Luciferase-Based Assay for the Discovery of Therapeutics That Prevent Malaria.

Author information

1
School of Medicine, Department of Pediatrics, Pharmacology & Drug Discovery, University of California, San Diego , 9500 Gilman Drive, La Jolla, California 92093, United States.
2
The Broad Institute, 415 Main Street, Cambridge, Massachusetts 02142, United States.
3
Genomics Institute of the Novartis Research Foundation (GNF) , 10675 John Jay Hopkins Drive, San Diego, California 92121, United States.
4
Department of Parasitology, Biomedical Primate Research Centre , P.O. Box 3306, 2280 GH Rijswijk, The Netherlands.
5
Medicines for Malaria Venture (MMV) , Meyrin 2015, Switzerland.

Abstract

In order to identify the most attractive starting points for drugs that can be used to prevent malaria, a diverse chemical space comprising tens of thousands to millions of small molecules may need to be examined. Achieving this throughput necessitates the development of efficient ultra-high-throughput screening methods. Here, we report the development and evaluation of a luciferase-based phenotypic screen of malaria exoerythrocytic-stage parasites optimized for a 1536-well format. This assay uses the exoerythrocytic stage of the rodent malaria parasite, Plasmodium berghei, and a human hepatoma cell line. We use this assay to evaluate several biased and unbiased compound libraries, including two small sets of molecules (400 and 89 compounds, respectively) with known activity against malaria erythrocytic-stage parasites and a set of 9886 diversity-oriented synthesis (DOS)-derived compounds. Of the compounds screened, we obtain hit rates of 12-13 and 0.6% in preselected and naïve libraries, respectively, and identify 52 compounds with exoerythrocytic-stage activity less than 1 μM and having minimal host cell toxicity. Our data demonstrate the ability of this method to identify compounds known to have causal prophylactic activity in both human and animal models of malaria, as well as novel compounds, including some exclusively active against parasite exoerythrocytic stages.

KEYWORDS:

drug discovery; exoerythrocytic-stage malaria; high-throughput screening; liver-stage malaria; malaria

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