Identification of novel leishmanicidal molecules by virtual and biochemical screenings targeting Leishmania eukaryotic translation initiation factor 4A
| Autor: | Imen Bassoumi-Jamoussi, Guillaume Bouvier, Michael Nilges, Mourad Barhoumi, N. Kyle Tanner, Josette Banroques, Arnaud Blondel, Ikram Guizani, Nathan Desdouits, Khadija Essafi-Benkhadir, Hélène Munier-Lehmann, Emna Harigua-Souiai, Yosser Zina Abdelkrim, Ons Zakraoui |
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| Přispěvatelé: | Laboratoire d'Epidémiologie Moléculaire et de Pathologie Expérimentale Appliquée aux Maladies Infectieuses (LR11IPT04), Université Tunis El Manar (UTM)-Institut Pasteur de Tunis, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Bioinformatique structurale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Expression Génétique Microbienne (EGM (UMR_8261 / FRE_3630)), Institut de biologie physico-chimique (IBPC), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Faculté des Sciences de Bizerte, Université de Carthage - University of Carthage, Chimie et Biocatalyse, This work received financial support from the Institut Pasteur PTR program (grant PTR426), and partially from the Ministry of Higher Education and Research in Tunisia (LR00IPT04 & LR11IPT04, IG), MN received financial support from the Investissement d’Avenir bioinformatics programme (Grant Bip:Bip ANR-BINF-03-01) and BayCellS FP7-IDEAS-ERC 294809, NKT and JB are supported by a Centre National de la Recherche Scientifique (CNRS), HeliDEAD grant [ANR-13- BSV8-0009-01] from the Agence Nationale de la Recherche and the Initiative d’Excellence program from the French State [DYNAMO, ANR-11-LABX-0011-01]. EHS received support from the UNESCO-L’Oréal 'For Women in Science' international fellowship and the Institut Pasteur International Network (Calmette and Yersin programme). The non commercial compounds were obtained from Dr. Laïla El Kihel from Université de Caen de Basse-Normandie, Centre d’Études et de Recherche sur le Médicament de Normandie (CERMN), UFR des Sciences Pharmaceutiques, for the minimal costs of shipping fees. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript, ANR-10-BINF-03-01/10-BINF-0003,Bip:Bip,Bip:Bip(2010), ANR-13-BSV8-0009,HeliDEAD,Les ARN hélicases à boite DEAD: quels sont leurs rôles, leurs partenaires, leurs substrats ARN et comment elles fonctionnent.(2013), ANR-11-LABX-0011/11-LABX-0011,DYNAMO,Dynamique des membranes transductrices d'énergie : biogénèse et organisation supramoléculaire.(2011), European Project: 294809,EC:FP7:ERC,ERC-2011-ADG_20110310,BAYCELLS(2012), Université de Tunis El Manar (UTM)-Institut Pasteur de Tunis, Bioinformatique structurale - Structural Bioinformatics, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut de biologie physico-chimique (IBPC (FR_550)), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Faculté des Sciences de Bizerte [Université de Carthage], ANR-10-BINF-0003,Bip:Bip,Paradigme d'inference bayesienne pour la Biologie structurale in silico(2010), ANR-11-LABX-0011,DYNAMO,Dynamique des membranes transductrices d'énergie : biogénèse et organisation supramoléculaire.(2011), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris] |
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
Models
Molecular 0301 basic medicine Life Cycles [SDV]Life Sciences [q-bio] Drug Evaluation Preclinical Protozoology MESH: Parasitic Sensitivity Tests Biochemistry Chemical library MESH: Eukaryotic Initiation Factor-4A Database and Informatics Methods White Blood Cells chemistry.chemical_compound Parasitic Sensitivity Tests Animal Cells [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases Medicine and Health Sciences Leishmania infantum MESH: Inhibitory Concentration 50 Protozoans Leishmania Adenosine Triphosphatases biology Chemistry lcsh:Public aspects of medicine Eukaryota Adenosine triphosphatase Parasitic diseases MESH: Leishmania infantum Enzymes 3. Good health Molecular Docking Simulation Infectious Diseases Helicases MESH: Drug Evaluation Preclinical Protozoan Life Cycles Cellular Types Sequence Analysis MESH: Models Molecular Research Article Amastigotes lcsh:Arctic medicine. Tropical medicine Bioinformatics lcsh:RC955-962 Immune Cells Immunology Antiprotozoal Agents Research and Analysis Methods Microbiology Inhibitory Concentration 50 03 medical and health sciences Eukaryotic translation MESH: Adenosine Triphosphatases MESH: Molecular Docking Simulation Initiation factor Binding site MESH: Antiprotozoal Agents Amastigote Virtual screening Blood Cells Binding Sites Promastigotes Macrophages Phosphatases Organisms Public Health Environmental and Occupational Health Biology and Life Sciences Proteins lcsh:RA1-1270 Cell Biology biology.organism_classification RNA helicases Parasitic Protozoans 030104 developmental biology MESH: Binding Sites Sequence motif analysis Eukaryotic Initiation Factor-4A Enzymology [INFO.INFO-BI]Computer Science [cs]/Bioinformatics [q-bio.QM] Developmental Biology |
| Zdroj: | PLoS Neglected Tropical Diseases PLoS Neglected Tropical Diseases, Public Library of Science, 2018, 12 (1), pp.e0006160. ⟨10.1371/journal.pntd.0006160⟩ PLoS Neglected Tropical Diseases, 2018, 12 (1), pp.e0006160. ⟨10.1371/journal.pntd.0006160⟩ PLoS Neglected Tropical Diseases, Vol 12, Iss 1, p e0006160 (2018) |
| ISSN: | 1935-2727 1935-2735 |
| DOI: | 10.1371/journal.pntd.0006160⟩ |
| Popis: | Leishmaniases are neglected parasitic diseases in spite of the major burden they inflict on public health. The identification of novel drugs and targets constitutes a research priority. For that purpose we used Leishmania infantum initiation factor 4A (LieIF), an essential translation initiation factor that belongs to the DEAD-box proteins family, as a potential drug target. We modeled its structure and identified two potential binding sites. A virtual screening of a diverse chemical library was performed for both sites. The results were analyzed with an in-house version of the Self-Organizing Maps algorithm combined with multiple filters, which led to the selection of 305 molecules. Effects of these molecules on the ATPase activity of LieIF permitted the identification of a promising hit (208) having a half maximal inhibitory concentration (IC50) of 150 ± 15 μM for 1 μM of protein. Ten chemical analogues of compound 208 were identified and two additional inhibitors were selected (20 and 48). These compounds inhibited the mammalian eIF4I with IC50 values within the same range. All three hits affected the viability of the extra-cellular form of L. infantum parasites with IC50 values at low micromolar concentrations. These molecules showed non-significant toxicity toward THP-1 macrophages. Furthermore, their anti-leishmanial activity was validated with experimental assays on L. infantum intramacrophage amastigotes showing IC50 values lower than 4.2 μM. Selected compounds exhibited selectivity indexes between 19 to 38, which reflects their potential as promising anti-Leishmania molecules. Author summary Leishmaniases constitute a group of neglected parasitic diseases that inflict major burden on public health. Novel drugs and targets need to be identified since current therapies have adverse side effects. Herein, we focused on Leishmania infantum translation initiation factor 4A (LieIF), as a potential drug target. LieIF, a pivotal enzyme in the translation machinery, is also implicated in host-pathogen interactions. We modeled its 3D structure and identified two pockets, which were used in virtual screenings of a chemical compound library. Therefore, we selected and purchased 305 compounds. We established a reliable ATPase screening assay to test the molecules against the enzymatic activity of LieIF and its mammalian homologue. A promising hit was retained and further characterized. It inhibited both proteins but showed different kinetic properties. It was used as a basis to identify similar analogues and two additional inhibitors were identified. All three hits reduced the viability of the extracellular promastigote form of the parasite, but they had no significant cytotoxic effects on host cells. They also affected the viability of the intracellular amastigote form and reduced the macrophage infection. This selectivity is very promising and indicates that these inhibitors would constitute an avenue to develop strategies to fight leishmaniases. |
| Databáze: | OpenAIRE |
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