Anti-malaria drug discovery: high-throughput assay to identify inhibitors of the Plasmodium falciparum apicoplast DNA polymerase
Year: 2016
Type: other
Abstract: Infection by the parasite Plasmodium falciparum is the leading cause of malaria in humans. The parasite contains a plastid-like organelle called the apicoplast, which is essential for survival within the host. The apicoplast maintains its own genome, which must undergo replication and repair. apPOL has been identified as the putative apicoplast replicative DNA polymerase and as it shares no direct orthology to mammalian polymerases, P. falciparum apPOL is an attractive anti-malarial drug target. We have developed a fluorescence-based high-throughput DNA polymerase assay to screen for compounds that inhibit apPOL activity. Analysis of validation experiments indicates that the assay is statistically robust with a signal window of 7.90 and a Z’ factor of 0.71. A pilot screen of a 2880 compound library identified 62 possible inhibitors that cause at least 50% inhibition of polymerase activity. Additionally, a collection of 400 compounds called the Malaria Box was screened. This collection is composed of chemicals that are known to be active against P. falciparum but their targets and mechanism are unknown. Nine of these compounds were identified as possible inhibitors of P. falciparum apPOL using our high-throughput assay. The simplicity and robustness of the assay provides a solid platform for screening apicoplast polymerase inhibitors which could serve as lead compounds in the efforts to discover and produce anti-malaria drugs.
Institution: Iowa State University
Language: English
Topic: Malaria Research and Control
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W2725715430