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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High-Throughput Screening Assays
Leishmania
Inositol
Leishmaniasis
Yeasts
Public-Private Sector Partnerships
Insect Vectors
Neglected Diseases
Active Immunotherapy
Enzyme Inhibitors
Drug Discovery
Saccharomyces cerevisiae
Membrane Proteins
Parasites
Drug Therapy
Enzymes
Infection
Proteins

Cite this

Norcliffe, Jennifer L. ; Mina, John G. ; Alvarez, Emilio ; Cantizani, Juan ; de Dios-Anton, Francisco ; Colmenarejo, Gonzalo ; Gonzalez-Del Valle, Silva ; Marco, Maria ; Fiandor, Jose M. ; Martin, Julio J. ; Steel, Patrick G. ; Denny, Paul W. / Identifying inhibitors of the Leishmania inositol phosphorylceramide synthase with antiprotozoal activity using a yeast-based assay and ultra-high throughput screening platform. In: Scientific Reports. 2018 ; Vol. 8, No. 3938.
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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.",
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Norcliffe, JL, Mina, JG, Alvarez, E, Cantizani, J, de Dios-Anton, F, Colmenarejo, G, Gonzalez-Del Valle, S, Marco, M, Fiandor, JM, Martin, JJ, Steel, PG & Denny, PW 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, vol. 8, no. 3938. https://doi.org/10.1038/s41598-018-22063-9

Identifying inhibitors of the Leishmania inositol phosphorylceramide synthase with antiprotozoal activity using a yeast-based assay and ultra-high throughput screening platform. / Norcliffe, Jennifer L.; Mina, John G.; Alvarez, Emilio; Cantizani, Juan; de Dios-Anton, Francisco; Colmenarejo, Gonzalo; Gonzalez-Del Valle, Silva; Marco, Maria; Fiandor, Jose M.; Martin, Julio J.; Steel, Patrick G.; Denny, Paul W.

In: Scientific Reports, Vol. 8, No. 3938, 02.03.2018.

Research output: Contribution to journalArticle

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AU - Marco, Maria

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