背景:人鼻病毒3C蛋白酶(HRV-3Cpro)在病毒增殖中起着至关重要的作用,将其确立为抗病毒治疗的首要目标。然而,关于鉴定HRV-3Cpro抑制剂的研究仍然有限。
目的:本研究有两个主要目的:为了验证终点比色测定的有效性,以前由我们的团队开发,用于鉴定HRV-3Cpro的潜在抑制剂;其次,在药用植物中发现抑制酶活性的植物化学物质。
方法:鲁普莱维,一种众所周知的HRV-3Cpro抑制剂,用于验证比色测定。在此之后,我们对2532种植物化学物质进行了两步筛选,这导致了八种活性化合物的鉴定:芹菜素,Carnosol,绿原酸,山奈酚,木犀草素,槲皮素,迷迭香酸,还有Rutin.我们随后在体外评估了这些候选物。为了进一步研究最有希望的候选人的抑制潜力,即,鼠尾草酚和迷迭香酸,进行分子对接研究以分析它们与HRV-3Cpro的结合相互作用。
结果:我们先前开发的比色测定法可有效鉴定选择性抑制HRV-3Cpro的化合物。鼠尾草酚和迷迭香酸作为强效抑制剂出现,体外抑制HRV-3Cpro活性超过55%。我们的分析表明,鼠尾草酚和迷迭香酸通过竞争机制发挥抑制作用。分子对接证实了它们与酶活性位点的竞争性结合。
结论:鼠尾草酚和迷迭香酸在开发冷疗方面的潜力值得进一步研究。通过强调这些化合物是有效的HRV-3Cpro抑制剂,我们的研究提供了一个有希望的方法来发现植物化学抑制剂对蛋白酶从类似的病原体。
BACKGROUND: Human rhinovirus 3C protease (HRV-3Cpro) plays a crucial role in viral proliferation, establishing it as a prime target for antiviral therapy. However, research on identifying HRV-3Cpro inhibitors is still limited.
OBJECTIVE: This study had two primary objectives: first, to validate the efficacy of an end-point colorimetric assay, previously developed by our team, for identifying potential inhibitors of HRV-3Cpro; and second, to discover phytochemicals in medicinal plants that inhibit the enzyme\'s activity.
METHODS: Rupintrivir, a well-known inhibitor of HRV-3Cpro, was used to validate the colorimetric assay. Following this, we conducted a two-step in silico screening of 2532 phytochemicals, which led to the identification of eight active compounds: apigenin, carnosol, chlorogenic acid, kaempferol, luteolin, quercetin, rosmarinic acid, and rutin. We subsequently evaluated these candidates in vitro. To further investigate the inhibitory potential of the most promising candidates, namely, carnosol and rosmarinic acid, molecular docking studies were performed to analyze their binding interactions with HRV-3Cpro.
RESULTS: The colorimetric assay we previously developed is effective in identifying compounds that selectively inhibit HRV-3Cpro. Carnosol and rosmarinic acid emerged as potent inhibitors, inhibiting HRV-3Cpro activity in vitro by over 55%. Our analysis indicated that carnosol and rosmarinic acid exert their inhibitory effects through a competitive mechanism. Molecular docking confirmed their competitive binding to the enzyme\'s active site.
CONCLUSIONS: Carnosol and rosmarinic acid warrant additional investigation for their potential in the development of common cold treatment. By highlighting these compounds as effective HRV-3Cpro inhibitors, our study presents a promising approach for discovering phytochemical inhibitors against proteases from similar pathogens.