| Congresso Brasileiro de Microbiologia 2023 | Resumo: 1162-1 | ||||
Resumo:Tuberculosis (TB) continues to persist as a formidable challenge in the global health landscape, standing out as one of the deadliest infectious diseases caused by a single pathogen. Effective TB treatment is hampered by multiple factors, primarily due to poor adherence and completion of treatments, largely attributed to their prolonged duration and substantial side effects. In this scenario, Plectasin, a fungal defensin isolated from Pseudopectania nigrella, represents a new beacon of hope. This compound has demonstrated its efficacy in eliminating strains of Mycobacterium tuberculosis (MTB) and multiple drug-resistant MTB strains. The primary aim of this research was to develop more compact and potent analogs of Plectasin's antimicrobial peptide (AMP), capable of eradicating MTB without causing adverse effects. To achieve this goal, the original AMP sequence, composed of 40 amino acids organized into a beta-sheet and an alpha-helix, was fragmented. Using machine learning-based bioinformatic analysis, it was determined that the anti-MTB activity resided in the beta-sheet. From this finding, permutations were generated in the cysteines and glycines of the beta-sheet, resulting in a library of 1102 AMP analogs. Applying bioinformatic techniques, we selected the four most promising analogs. Selection was based on their antimycobacterial activity in silico (AntiTbPpred), pharmacokinetic properties (pkcsm), half-life (plifepred), and concluded with molecular docking assays between the receptors and the AMP analogs (Vina). Subsequently, the four most promising analogs were synthesized using the F-moc solid-phase synthesis method. The synthesized products underwent LC/MS analysis and were purified using HPLC. Subsequently, cytotoxicity assays were conducted on MRC-5 lung fibroblasts and J774A.1 macrophages. Minimum inhibitory concentrations were determined using the Resazurin Microtiter Assay on the H37Rv strain. The obtained results demonstrated potent anti-MTB activity without inducing cytotoxicity in any of the cell lines, even at concentrations exceeding 100 µg/ml. This study highlights the immense potential of bioinformatics tools for the identification and development of new drugs, achieving a significant reduction in the costs and times of the drug discovery process. Palavras-chave: Antimicrobial peptide, Bioinformatic, Drug discovery , Tuberculosis Agência de fomento:CAPES | |||||