结核病(TB)是由结核分枝杆菌(Mtb)引起的传染病,这仍然是一个重大的全球卫生挑战。多药耐药(MDR)Mtb菌株的出现要求开发新的治疗策略。这项研究的重点是通过对内部化学图书馆的全面筛选来鉴定和评估针对MtbH37Ra的潜在抑制剂。随后,一种有前途的嘧啶衍生物(LQM495)被认为是有前途的,然后通过实验和计算机模拟方法进一步研究。在这种情况下,计算技术用于阐明LQM495抑制作用的潜在分子靶标。然后,共识反向对接(CRD)方案用于研究该化合物与几个Mtb靶标之间的相互作用.在调查的98个Mtb目标中,增强的细胞内存活(Eis)蛋白成为LQM495的靶标。要深入了解LQM495-Eis复合体的稳定性,分子动力学(MD)模拟在400ns的轨迹上进行。通过量子力学(QM)方法获得了对Eis结合位点内结合模式的进一步了解,使用密度泛函理论(DFT),与B3LYP/D3基础设置。这些计算揭示了LQM495的电子性质和反应性。随后,Eis活性的抑制试验和动力学研究用于研究LQM495的活性。然后,在Eis蛋白上发现LQM495的IC50值为11.0±1.4µM。此外,其Vmax,Km,和Ki参数表明它是竞争性抑制剂。最后,本研究提出LQM495作为一种有前途的MtbEis蛋白抑制剂,未来可以进一步探索开发新的抗结核药物。
Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (Mtb), which remains a significant global health challenge. The emergence of multidrug-resistant (MDR) Mtb strains imposes the development of new therapeutic strategies. This study focuses on the identification and evaluation of potential inhibitors against Mtb H37Ra through a comprehensive screening of an in-house chemolibrary. Subsequently, a promising pyrimidine derivative (LQM495) was identified as promising and then further investigated by experimental and in silico approaches. In this context, computational techniques were used to elucidate the potential molecular target underlying the inhibitory action of LQM495. Then, a consensus reverse docking (CRD) protocol was used to investigate the interactions between this compound and several Mtb targets. Out of 98 Mtb targets investigated, the enhanced intracellular survival (Eis) protein emerged as a target for LQM495. To gain insights into the stability of the LQM495-Eis complex, molecular dynamics (MD) simulations were conducted over a 400 ns trajectory. Further insights into its binding modes within the Eis binding site were obtained through a Quantum mechanics (
QM) approach, using density functional theory (DFT), with B3LYP/D3 basis set. These calculations shed light on the electronic properties and reactivity of LQM495. Subsequently, inhibition assays and kinetic studies of the Eis activity were used to investigate the activity of LQM495. Then, an IC50 value of 11.0 ± 1.4 µM was found for LQM495 upon Eis protein. Additionally, its Vmax, Km, and Ki parameters indicated that it is a competitive inhibitor. Lastly, this study presents LQM495 as a promising inhibitor of Mtb Eis protein, which could be further explored for developing novel anti-TB drugs in the future.