ABSTRACT
Aim: A structural model of chorismate synthase (CS) from the pathogenic fungus Candida albicans was used for virtual screening simulations. Methods: Docking, molecular dynamics, cell growth inhibition and protein binding assays were used for search and validation. Results: Two molecules termed CS8 and CaCS02 were identified. Further studies of the minimal inhibitory concentration demonstrated fungicidal activity against Paracoccidioides brasiliensis with a minimal inhibitory concentration and minimal fungicidal concentration of 512 and 32 µg·ml-1 for CS8 and CaCS02, respectively. In addition, CaCS02 showed a strong synergistic effect in combination with amphotericin B without cytotoxic effects. In vitro studies using recombinant CS from P. brasiliensis showed IC50 of 29 µM for CaCS02 supporting our interpretation that inhibition of CS causes the observed fungicidal activity.
Subject(s)
Antifungal Agents/pharmacology , Fungal Proteins/antagonists & inhibitors , Paracoccidioides/drug effects , Phosphorus-Oxygen Lyases/antagonists & inhibitors , Amino Acid Sequence , Amphotericin B/pharmacology , Animals , Antifungal Agents/chemistry , Antifungal Agents/metabolism , Candida albicans/enzymology , Chlorocebus aethiops , Drug Synergism , Fungal Proteins/chemistry , Fungal Proteins/metabolism , HeLa Cells , Humans , Inhibitory Concentration 50 , Microbial Sensitivity Tests , Molecular Docking Simulation , Molecular Structure , Paracoccidioides/enzymology , Phosphorus-Oxygen Lyases/chemistry , Phosphorus-Oxygen Lyases/metabolism , Protein Binding , Vero CellsABSTRACT
Aim: 17 new 4-methoxynaphthalene-N-acylhydrazones were synthesized in order to evaluate their biological action against important pathogens. Methods: In vitro susceptibility assays of compounds were performed against Paracoccidioidesbrasiliensis and Mycobacterium tuberculosis. Results: Compounds 4a, 4b and 4k were the most potent against P. brasiliensis, two with minimum inhibitory concentrations of ≤1 µg ml-1 and exhibited pharmacological synergy with amphotericin B. The compounds also showed activity against M. tuberculosis, with 4c and 4k being the more promising. Compound 4k showed good synergistic antimycobacterium activity with ethambutol. None of the compounds tested showed toxicity. Conclusion: We highlight the compound 4k, as a potential agent for the treatment of patients co-infected with paracoccidioidomycosis and tuberculosis.
Subject(s)
Anti-Bacterial Agents/pharmacology , Antifungal Agents/pharmacology , Coinfection/drug therapy , Mycobacterium tuberculosis/drug effects , Paracoccidioides/drug effects , Paracoccidioidomycosis/drug therapy , Tuberculosis/drug therapy , Amphotericin B/pharmacology , Anti-Bacterial Agents/chemical synthesis , Antifungal Agents/chemical synthesis , Drug Combinations , Drug Discovery , Drug Synergism , Ethambutol/pharmacology , Humans , Microbial Sensitivity Tests , Mycobacterium tuberculosis/pathogenicity , Paracoccidioides/pathogenicityABSTRACT
AIM: We investigated a proteome profile, protein-protein interaction and morphological changes of Mycobacterium tuberculosis after different times of eupomatenoid-5 (EUP-5) induction to evaluate the cellular response to the drug-induced damages. METHODS: The bacillus was induced to sub-minimal inhibitory concentration of EUP-5 at 12 h, 24 h and 48 h. The proteins were separated by 2D gel electrophoresis, identified by LC/MS-MS. Scanning electron microscopy and Search Tool for the Retrieval of Interacting Genes/Proteins analyses were performed. RESULTS: EUP-5 impacts mainly in M. tuberculosis proteins of intermediary metabolism and interactome suggests a multisite disturbance that contributes to bacilli death. Scanning electron microscopy revealed the loss of bacillary form. CONCLUSION: Some of the differentially expressed proteins have the potential to be drug targets such as citrate synthase (Rv0896), phosphoglycerate kinase (Rv1437), ketol-acid reductoisomerase (Rv3001c) and ATP synthase alpha chain (Rv1308).
