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Functional, thermodynamics, structural and biological studies of in silico-identified inhibitors of Mycobacterium tuberculosis enoyl-ACP(CoA) reductase enzyme

2017; Nature Portfolio; Volume: 7; Issue: 1 Linguagem: Inglês

10.1038/srep46696

2045-2322

Leonardo Martinelli, Mariane Rotta, Anne Drumond Villela, Valnês S. Rodrigues-Junior, Bruno Lopes Abbadi, Rogério Valim Trindade, Guilherme Oliveira Petersen, Giuliano Machado Danesi, Laura Roesler Nery, Ivaní Pauli, Maria M. Campos, Carla Denise Bonan, Osmar Norberto de Souza, Luiz Augusto Basso, Diógenes Santiago Santos,

Antibiotic Resistance in Bacteria

Abstract Novel chemotherapeutics agents are needed to kill Mycobacterium tuberculosis , the main causative agent of tuberculosis (TB). The M. tuberculosis 2- tran s-enoyl-ACP(CoA) reductase enzyme ( Mt InhA) is the druggable bona fide target of isoniazid. New chemotypes were previously identified by two in silico approaches as potential ligands to Mt InhA. The inhibition mode was determined by steady-state kinetics for seven compounds that inhibited Mt InhA activity. Dissociation constant values at different temperatures were determined by protein fluorescence spectroscopy. van’t Hoff analyses of ligand binding to Mt InhA:NADH provided the thermodynamic signatures of non-covalent interactions (Δ H °, Δ S °, Δ G °). 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 Mt InhA:NADH:Labio_16 ternary complex. The results show that the Labio_16 compound is a direct inhibitor of Mt InhA, and it may represent a hit for the development of chemotherapeutic agents to treat TB.