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
| العنوان: | 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 |
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| المؤلفون: | Laurent Meijer, Gerald F. Späth, Alexios-Leandros Skaltsounis, Nicolas Gaboriaud-Kolar, Nathalie Aulner, Olivier Leclercq, Konstantina Vougogiannopoulou, Audrey Defontaine, Nassima Oumata, Hervé Galons, Emilie Durieu, Eric Prina, Najma Rachidi, Sandrine Ruchaud, Joo Hwan No |
| المساهمون: | 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), ManRos Therapeutics, Department of Pharmacognosy and Chemistry of Natural Products, National and Kapodistrian University of Athens (NKUA), Imagopole (CITECH), 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 du CNRS (INC)-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), Institut Pasteur [Paris], Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur [Paris], Parasitologie moléculaire et Signalisation, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), National and Kapodistrian University of Athens = University of Athens (NKUA | UoA), Institut Pasteur de Corée, Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS), 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-0062/10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), ANR-10-INBS-04-01/10-INBS-0004,France-BioImaging,Développment d'une infrastructure française distribuée coordonnée(2010), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Station biologique de Roscoff [Roscoff] (SBR), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS) |
| المصدر: | Antimicrobial Agents and Chemotherapy Antimicrobial Agents and Chemotherapy, 2016, ⟨10.1128/AAC.00021-16⟩ Antimicrobial Agents and Chemotherapy, American Society for Microbiology, 2016, ⟨10.1128/AAC.00021-16⟩ |
| بيانات النشر: | HAL CCSD, 2016. |
| سنة النشر: | 2016 |
| مصطلحات موضوعية: | 0301 basic medicine, 030106 microbiology, Antiprotozoal Agents, Biology, Cell Line, 03 medical and health sciences, Casein Kinase I, Drug Discovery, Animals, Humans, Protein Isoforms, [CHIM]Chemical Sciences, Pharmacology (medical), Amastigote, Leishmania, Pharmacology, Kinase, Drug discovery, Intracellular parasite, Macrophages, Biological activity, biology.organism_classification, 3. Good health, 030104 developmental biology, Infectious Diseases, Biochemistry, Casein kinase 1 |
| الوصف: | 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 exoprotein 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 antileishmanial compounds. In step 1, we screened 5,018 compounds from kinase-biased libraries with Leishmania and mammalian CK1 in order to identify hit compounds and assess their specificity. For step 2, we selected 88 compounds among those with the lowest 50% inhibitory concentration 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 75 compounds showed antileishmanial 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 antileishmanial activity. |
| اللغة: | English |
| تدمد: | 0066-4804 1098-6596 |
| DOI: | 10.1128/AAC.00021-16⟩ |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::43bb94a0e1dc335bc918aab047cec35e https://hal.sorbonne-universite.fr/hal-01299756/document |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....43bb94a0e1dc335bc918aab047cec35e |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 00664804 10986596 |
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| DOI: | 10.1128/AAC.00021-16⟩ |