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

Jennifer L. Norcliffe, John G. Mina, Emilio Alvarez, Juan Cantizani, Francisco de Dios-Anton, Gonzalo Colmenarejo, Silva Gonzalez-Del Valle, Maria Marco, Jose M. Fiandor, Julio J. Martin, Patrick G. Steel, Paul W. Denny

    Research output: Contribution to journalArticle

    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.
    Original languageEnglish
    Number of pages10
    JournalScientific Reports
    Volume8
    Issue number3938
    DOIs
    Publication statusPublished - 2 Mar 2018

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    Norcliffe, J. L., Mina, J. G., Alvarez, E., Cantizani, J., de Dios-Anton, F., Colmenarejo, G., Gonzalez-Del Valle, S., Marco, M., Fiandor, J. M., Martin, J. J., Steel, P. G., & Denny, P. W. (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(3938). https://doi.org/10.1038/s41598-018-22063-9