Núcleo de Pesquisa Em Microbiologia Médica, Universidade Federal de Rio Grande, Rua General Osório s/n, Rio Grande, RS, Brazil.
Núcleo de Pesquisa Em Microbiologia Médica, Universidade Federal de Rio Grande, Rua General Osório s/n, Rio Grande, RS, Brazil.
Tuberculosis (Edinb). 2019 Sep;118:101853. doi: 10.1016/j.tube.2019.07.004. Epub 2019 Jul 23.
Herein, we evaluated tetrahydropyridine (THP) compounds (NUNM) as antimicrobials and inhibitors of the efflux mechanism in M. abscessus. subsp. abscessus. The modulation factor (MF) of efflux inhibitors was calculated from the minimum inhibitory concentrations (MICs) of amikacin (AMI), ciprofloxacin (CIP) and clarithromycin (CLA) in the absence and presence of subinhibitory concentrations of the NUNM compounds and canonical inhibitors carbonyl cyanide m-chlorophenyl hydrazone (CCCP) and verapamil (VP). The kinetics of the intracellular accumulation of the fluorimetric substrate ethidium bromide (EtBr) was evaluated and calculated by the relative final fluorescence (RFF). In addition, molecular modeling simulations for the MmpL5 and Tap efflux transporters with ligands (CLA, NUNM, CCCP, VP and EtBr) were performed to better understand the efflux mechanism. We highlight the NUNM01 compound because it reduced the MICs of AMI, CIP and CLA by 4-, 4- and 16-fold, respectively, had the highest effect on EtBr accumulation (RFF = 3.1) and showed a significant in silico affinity for the evaluated proteins in docking simulations. Based on the analyses performed in vitro and in silico, we propose that NUNM01 is a potential pharmacophore candidate for the development of a therapeutic adjuvant for M. abscessus infections.
在此，我们评估了四氢吡啶（THP）化合物（NUNM）作为分枝杆菌属亚种脓肿的抗菌剂和外排机制抑制剂。从阿米卡星（AMI）、环丙沙星（CIP）和克拉霉素（CLA）的最小抑菌浓度（MIC）中计算出外排抑制剂的调制因子（MF），这些 MIC 值是在存在和不存在亚抑菌浓度的 NUNM 化合物和经典抑制剂羰基氰化物 m-氯苯腙（CCCP）和维拉帕米（VP）的情况下得出的。通过相对最终荧光（RFF）评估和计算了荧光底物溴化乙锭（EtBr）的细胞内积累动力学。此外，还对 MmpL5 和 Tap 外排转运蛋白与配体（CLA、NUNM、CCCP、VP 和 EtBr）进行了分子建模模拟，以更好地理解外排机制。我们重点介绍 NUNM01 化合物，因为它使 AMI、CIP 和 CLA 的 MIC 分别降低了 4 倍、4 倍和 16 倍，对 EtBr 积累的影响最大（RFF=3.1），并且在对接模拟中显示出对评估蛋白的显著计算亲和力。基于体外和计算分析，我们提出 NUNM01 是开发脓肿分枝杆菌感染治疗佐剂的潜在药效团候选物。
