Amoebae-Based Screening Reveals a Novel Family of Compounds Restricting Intracellular <i>Legionella pneumophila</i>

Article Article scientifique Article Amoebae-Based Screening Reveals a Novel Family of Compounds Restricting Intracellular <i>Legionella pneumophila</i> eng 2015 author Harrison Christopher F. author Chiriano Gianpaolo 784524 author Finsel Ivo author Manske Christian author Hoffmann Christine author Steiner Bernhard author Kranjc Pietrucci Agata 712787 author Patthey Ophelie 135760 author Kicka Sébastien 736074 author Trofimov Valentin 709788 author Ouertatani Hajer 734371 author Soldati Thierry 0000-0002-2056-7931 135632 author Scapozza Leonardo 600318 author Hilbi Hubert 327-338 10.1021/acsinfecdis.5b00002 2373-8227 The causative agent of Legionnaires' disease, Legionella pneumophila, grows in environmental amoebae and mammalian macrophages within a distinct compartment, the ‘Legionella-containing vacuole' (LCV). Intracellular bacteria are protected from many antibiotics, and thus are notoriously difficult to eradicate. To identify novel compounds that restrict intracellular bacterial replication, we previously developed an assay based on a coculture of amoebae and GFP-producing L. pneumophila. This assay was used to screen a pathway-based, highly diverse chemical library, referred to as the Sinergia library. In this work, we chose to focus on a group of 11 hit compounds, the majority of which originated from the query molecule CN585, a compound that targets the protein phosphatase calcineurin. Further studies on 78 related compound variants revealed crucial structural attributes, namely a triple-ring scaffold with a central triazine moiety, substituted in positions 3 and 5 by two piperidine or pyrrolidine rings, and in position 1 by an amine group bearing a single aliphatic chain moiety. The most effective compound, ZINC00615682, inhibited intracellular replication of L. pneumophila with an IC50 of approximately 20 nM in Acanthamoeba castellanii and slightly less efficiently in Dictyostelium discoideum or macrophages. Pharmacological and genetic attempts to implicate calcineurin in the intracellular replication of L. pneumophila failed. Taken together, these results show that the amoebae-based screen and structure–activity relationship analysis is suitable for the identification of novel inhibitors of the intracellular replication of L. pneumophila. The most potent compound identified in this study targets (an) as yet unidentified host factor(s). Amoeba Antibiotics Antivirulence Calcineurin Intracellular replication Legionella Macrophage Pathogen vacuole Screen Structure−activity relationship (SAR) Type IV secretion Dewey 540 Dewey 612 Dewey 615 39 Département de biochimie 144 31 Section des sciences pharmaceutiques 17 134 Département de physiologie cellulaire et métabolisme 21C false 766 Groupe Soldati ACS infectious diseases 1 7 4a5d613a-a037-4c61-b24c-43547132d3c3 Article (Published version) Public acsinfecdis%2E5b00002(1).pdf 1932066 application/pdf openAccess Faculté des sciences / Section de chimie et biochimie / Département de biochimie Faculté des sciences / Section des sciences pharmaceutiques Faculté de médecine / Section de médecine fondamentale / Département de physiologie cellulaire et métabolisme

Title	Published in	Year	Views	Downloads

Amoebae-Based Screening Reveals a Novel Family of Compounds Restricting Intracellular Legionella pneumophila	ACS infectious diseases	2015	694	554
Melatonin improves muscle function of the dystrophic mdx(5Cv) mouse, a model for Duchenne muscular dystrophy	Journal of pineal research	2011	645	5

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