Functional, thermodynamics, structural and biological studies of in silico-identified inhibitors of Mycobacterium tuberculosis enoyl-ACP(CoA) reductase enzyme
| Autor: | Mariane Rotta, Giuliano Machado Danesi, Bruno Lopes Abbadi, Ivani Pauli, Carla Denise Bonan, Osmar Norberto de Souza, Guilherme Oliveira Petersen, Diógenes Santiago Santos, Laura Roesler Nery, Rogério Valim Trindade, Valnês S. Rodrigues-Junior, Leonardo K. Martinelli, Maria M. Campos, Luiz Augusto Basso, Anne Drumond Villela |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
In silico Antitubercular Agents Microbial Sensitivity Tests Reductase Article Cell Line Mycobacterium tuberculosis Mice 03 medical and health sciences Bacterial Proteins Chlorocebus aethiops medicine Animals Humans Tuberculosis Computer Simulation Enzyme Inhibitors Vero Cells Ternary complex chemistry.chemical_classification Multidisciplinary 030102 biochemistry & molecular biology biology Chemistry Isoniazid biology.organism_classification zebrafish Dissociation constant Kinetics HaCaT RAW 264.7 Cells 030104 developmental biology Enzyme Biochemistry Thermodynamics Oxidoreductases medicine.drug |
| Zdroj: | Repositório Institucional PUCRS Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS) instacron:PUC_RS Scientific Reports |
| Popis: | Novel chemotherapeutics agents are needed to kill Mycobacterium tuberculosis, the main causative agent of tuberculosis (TB). The M. tuberculosis 2-trans-enoyl-ACP(CoA) reductase enzyme (MtInhA) is the druggable bona fide target of isoniazid. New chemotypes were previously identified by two in silico approaches as potential ligands to MtInhA. The inhibition mode was determined by steady-state kinetics for seven compounds that inhibited MtInhA activity. Dissociation constant values at different temperatures were determined by protein fluorescence spectroscopy. van’t Hoff analyses of ligand binding to MtInhA:NADH provided the thermodynamic signatures of non-covalent interactions (ΔH°, ΔS°, ΔG°). Phenotypic screening showed that five compounds inhibited in vitro growth of M. tuberculosis H37Rv strain. Labio_16 and Labio_17 compounds also inhibited the in vitro growth of PE-003 multidrug-resistant strain. Cytotoxic effects on Hacat, Vero and RAW 264.7 cell lines were assessed for the latter two compounds. The Labio_16 was bacteriostatic and Labio_17 bactericidal in an M. tuberculosis-infected macrophage model. In Zebrafish model, Labio_16 showed no cardiotoxicity whereas Labio_17 showed dose-dependent cardiotoxicity. Accordingly, a model was built for the MtInhA:NADH:Labio_16 ternary complex. The results show that the Labio_16 compound is a direct inhibitor of MtInhA, and it may represent a hit for the development of chemotherapeutic agents to treat TB. |
| Databáze: | OpenAIRE |
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