title: From Drug Screening to Target Deconvolution: a Target-Based Drug Discovery Pipeline Using Leishmania Casein Kinase 1 Isoform 2 To Identify Compounds with Antileishmanial Activity.
abstract: 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.
authors.affiliation: Sorbonne Universités, UPMC Université de Paris 06, CNRS USR3151, Protein Phosphorylation and Human Diseases, Station Biologique, Roscoff, France.
authors.forename: Emilie
authors.initials: E
authors.lastname: Durieu
authors.affiliation: Institut Pasteur, Laboratoire d'Immunophysiologie et Parasitisme, Paris, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Prina
authors.affiliation: Institut Pasteur and INSERM U1201, Unité de Parasitologie moléculaire et Signalisation, Paris, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Leclercq
authors.affiliation: ManRos Therapeutics, Centre de Perharidy, Roscoff, France.
authors.forename: Nassima
authors.initials: N
authors.lastname: Oumata
authors.affiliation: Department of Pharmacognosy and Natural Products Chemistry, School of Pharmacy, University of Athens, Athens, Greece.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Gaboriaud-Kolar
authors.affiliation: Department of Pharmacognosy and Natural Products Chemistry, School of Pharmacy, University of Athens, Athens, Greece.
authors.forename: Konstantina
authors.initials: K
authors.lastname: Vougogiannopoulou
authors.affiliation: Institut Pasteur, Imagopole, Paris, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Aulner
authors.affiliation: Sorbonne Universités, UPMC Université de Paris 06, CNRS USR3151, Protein Phosphorylation and Human Diseases, Station Biologique, Roscoff, France.
authors.forename: Audrey
authors.initials: A
authors.lastname: Defontaine
authors.affiliation: Institut Pasteur Korea, Leishmania Research Laboratory, Seongnam-si, Republic of Korea.
authors.forename: Joo Hwan
authors.initials: JH
authors.lastname: No
authors.affiliation: Sorbonne Universités, UPMC Université de Paris 06, CNRS USR3151, Protein Phosphorylation and Human Diseases, Station Biologique, Roscoff, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Ruchaud
authors.affiliation: Department of Pharmacognosy and Natural Products Chemistry, School of Pharmacy, University of Athens, Athens, Greece.
authors.forename: Alexios-Leandros
authors.initials: AL
authors.lastname: Skaltsounis
authors.affiliation: Université Paris-Descartes, Laboratoire de Chimie Organique 2, INSERM U 648, Paris, France.
authors.forename: Hervé
authors.initials: H
authors.lastname: Galons
authors.affiliation: Institut Pasteur and INSERM U1201, Unité de Parasitologie moléculaire et Signalisation, Paris, France.
authors.forename: Gerald F
authors.initials: GF
authors.lastname: Späth
authors.affiliation: Sorbonne Universités, UPMC Université de Paris 06, CNRS USR3151, Protein Phosphorylation and Human Diseases, Station Biologique, Roscoff, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Meijer
authors.affiliation: Sorbonne Universités, UPMC Université de Paris 06, CNRS USR3151, Protein Phosphorylation and Human Diseases, Station Biologique, Roscoff, France najma.rachidi@pasteur.fr.
authors.forename: Najma
authors.initials: N
authors.lastname: Rachidi
mesh_terms: D000818:Animals; D000981:Antiprotozoal Agents / Q000737:chemistry / Q000494:pharmacology*; D047389:Casein Kinase I / Q000378:metabolism; D002460:Cell Line; D055808:Drug Discovery; D006801:Humans; D007891:Leishmania / Q000187:drug effects* / Q000378:metabolism; D008264:Macrophages / Q000469:parasitology; D020033:Protein Isoforms / Q000378:metabolism
chemical_list: D000981:Antiprotozoal Agents; D020033:Protein Isoforms; D047389:Casein Kinase I