Identifying inhibitors of the Leishmania inositol phosphorylceramide synthase with antiprotozoal activity using a yeast-based assay and ultra-high throughput screening platform

Norcliffe, J.L.; Mina, J.G.; Alvarez-Ruiz, E.; Cantizani-Perez, J.; de Dios-Anton, F.; Colmenarejo, G.; Gonzalez-Del Valle, S.; Marco-Martin, M.; Fiandor, J.M.; Martin-Plaza, J.J.; Steel, P.G.; Denny, P.W.

Abstract

Leishmaniasis is a Neglected Tropical Disease caused by the insect-vector borne protozoan parasite, Leishmania species. Infection affects millions of the world’s poorest, however vaccines are absent and drug therapy limited. Recently, public-private partnerships have developed to identify new modes of controlling leishmaniasis. Drug discovery is a significant part of these efforts and here we describe the development and utilization of a novel assay to identify antiprotozoal inhibitors of the Leishmania enzyme, inositol phosphorylceramide (IPC) synthase. IPC synthase is a membrane-bound protein with multiple transmembrane domains, meaning that a conventional in vitro assay using purified protein in solution is highly challenging. Therefore, we utilized Saccharomyces cerevisiae as a vehicle to facilitate ultra-high throughput screening of 1.8 million compounds. Antileishmanial benzazepanes were identified and shown to inhibit the enzyme at nanomolar concentrations. Further chemistry produced a benzazepane that demonstrated potent and specific inhibition of IPC synthase in the Leishmania cell.

Citation

Norcliffe, J., Mina, J., Alvarez-Ruiz, E., Cantizani-Perez, J., de Dios-Anton, F., Colmenarejo, G., …Denny, P. (2018). Identifying inhibitors of the Leishmania inositol phosphorylceramide synthase with antiprotozoal activity using a yeast-based assay and ultra-high throughput screening platform. Scientific Reports, 8, Article 3938. https://doi.org/10.1038/s41598-018-22063-9

• Journal Article Type: Article
• Acceptance Date: Feb 16, 2018
• Online Publication Date: Mar 2, 2018
• Publication Date: Mar 2, 2018
• Deposit Date: Jan 8, 2018
• Publicly Available Date: Mar 2, 2018
• Journal: Scientific Reports
• Publisher: Nature Research
• Peer Reviewed: Peer Reviewed
• Volume: 8
• Article Number: 3938
• DOI: https://doi.org/10.1038/s41598-018-22063-9
• Public URL: https://durham-repository.worktribe.com/output/1337242