From drug screening to target deconvolution: A target-based drug discovery pipeline using Leishmania casein kinase 1 isoform 2 to identify compounds with anti-leishmanial activity
In: ISSN: 0066-4804, 2016
Online
academicJournal
Zugriff:
International audience ; Existing therapies for leishmaniases present significant limitations, such as toxic side effects, and are rendered inefficient by parasite resistance. It is of utmost importance to develop novel drugs targeting Leishmania that take these two limitations into consideration. We thus chose a target-based approach using an exo-protein kinase, Leishmania casein kinase 1.2 (LmCK1.2) that was recently shown to be essential for intracellular parasite survival and infectivity. We developed a four-step pipeline to identify novel selective anti-leishmanial compounds. In step 1, we screened 5018 compounds from kinase-biased libraries with Leishmania and mammalian casein kinase 1 in order to identify hit compounds and assess their specificity. For step 2, we selected 88 compounds among those with the lowest IC50 to test their biological activity on host-free parasites using a resazurin reduction assay and on intramacrophagic amastigotes using a high content phenotypic assay. Only seventy-five compounds showed anti-leishmanial activity and were retained for step 3 to evaluate their toxicity against mouse macrophages and human cell lines. The four compounds that displayed a selectivity index above 10 were then assessed for their affinity to LmCK1.2 using a target deconvolution strategy in step 4. Finally, we retained two compounds, PP2 and compound 42, for which LmCK1.2 seems to be the primary target. Using this four-step pipeline, we identify from several thousand molecules, two lead compounds with a selective anti-leishmanial activity.
Titel: |
From drug screening to target deconvolution: A target-based drug discovery pipeline using Leishmania casein kinase 1 isoform 2 to identify compounds with anti-leishmanial activity
|
---|---|
Autor/in / Beteiligte Person: | Durieu, Emilie ; Prina, Eric ; Leclercq, Olivier ; Oumata, Nassima ; Gaboriaud-Kolar, Nicolas ; Vougogiannopoulou, Konstantina ; Aulner, Nathalie ; Defontaine, Audrey ; No, Joo Hwan ; Ruchaud, Sandrine ; Skaltsounis, Alexios-Leandros ; Galons, Hervé ; Späth, Gerald F ; Meijer, Laurent ; Rachidi, Najma ; Phophorylation de protéines et Pathologies Humaines (P3H) ; Station biologique de Roscoff Roscoff (SBR) ; Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS) ; Immunophysiologie et Parasitisme Intracellulaire ; Institut Pasteur Paris (IP) ; Parasitologie moléculaire et Signalisation / Molecular Parasitology and Signaling ; Institut Pasteur Paris (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM) ; Therapeutics, ManRos ; Department of Pharmacognosy and Chemistry of Natural Products ; National and Kapodistrian University of Athens (NKUA) ; (CITECH), Imagopole ; Institut Pasteur Korea - Institut Pasteur de Corée ; Réseau International des Instituts Pasteur (RIIP) ; Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601) ; Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie - CNRS Chimie (INC-CNRS)-Centre National de la Recherche Scientifique (CNRS) ; This study was supported by the 7th Framework Program of the European Commission through grants to the LEISHDRUG project (223414), by ANR-11-RPIB-0016 TRANSLEISH, and by the French Government's Investissements d'Avenir Program Laboratoire d'Excellence Integrative Biology of Emerging Infectious Diseases (grant ANR-10-LABX-62-IBEID). The Imagopole-CiTech is part of the FranceBioImaging infrastructure supported by the French National Research Agency (ANR-10-INSB-04-01, Investments for the Future) and is supported by the Conseil de la Region Ile-de-France (program Sesame 2007, project Imagopole, S. Shorte) and the Fondation Française pour la Recherche Médicale (Programme Grands Equipements N.A. ) ; ANR-11-RPIB-0016,TRANSLEISH,Découverte de nouvelles protéines kinases chez Leishmania donovani à partir des inhibiteurs tête de série issus d'un criblage phénotypique(2011) ; ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010) ; ANR-10-INBS-0004,France-BioImaging,Développment d'une infrastructure française distribuée coordonnée(2010) ; European Project: 223414,EC:FP7:HEALTH,FP7-HEALTH-2007-B,LEISHDRUG(2008) |
Link: | |
Zeitschrift: | ISSN: 0066-4804, 2016 |
Veröffentlichung: | HAL CCSD ; American Society for Microbiology, 2016 |
Medientyp: | academicJournal |
DOI: | 10.1128/AAC.00021-16 |
Schlagwort: |
|
Sonstiges: |
|