Mastalipour Mohammadamin, Gering Ian, Coronado Mônika Aparecida, González Jorge Enrique Hernández, Willbold Dieter, Eberle Raphael Josef
Institut für Physikalische Biologie, Heinrich-Heine-Universität Düsseldorf, Universitätsstraße 1, 40225 Düsseldorf, Germany.
Institut für Biologische Informationsprozesse, Strukturbiochemie (IBI-7), Forschungszentrum Jülich, 52425 Jülich, Germany.
Curr Res Microb Sci. 2025 Mar 11;8:100376. doi: 10.1016/j.crmicr.2025.100376. eCollection 2025.
Chikungunya virus (CHIKV) is an emerging pathogen affecting populations worldwide, with rapidly increasing infection rates. CHIKV, an arbovirus of the alphavirus genus, is predominantly found in tropical regions and transmitted by mosquitoes. Climate change has accelerated the global spread of these vectors, leading to outbreaks in non-tropical regions, including parts of Europe. The absence of antiviral therapies and the potential for co-infections with other viruses make CHIKV a significant public health concern. CHIKV replication relies on nsP2 cysteine protease activity to cleave its viral polyprotein into functional nonstructural and structural proteins. Targeting the nsP2 protease represents a promising strategy for antiviral therapy development. In this study, phage display was used to screen a library of peptides for potential binders of the target protease. Biophysical and biochemical analyses of the identified peptides assessed their inhibitory potential. Among the six identified peptides (named as P1-P6), four demonstrated inhibitory effects on the nsP2 protease (nsP2). Peptide P1 exhibited the strongest inhibitory effect, with a half-maximal inhibitory concentration (IC) of 4.6 ± 1.9 µM, and a low cytotoxicity. The secondary structure analysis through CD spectroscopy and homology modelling revealed that P1 adopts an alpha-helical conformation. Finally, molecular dynamics simulations enabled us to investigate the dynamics of the nsP2 active site and molecular docking was employed to predict the orthosteric binding mode of P1, providing insights into protein-peptide interaction. These findings underscore the potential of peptide P1 as a lead compound for further investigation in the context of CHIKV research.
基孔肯雅病毒（CHIKV）是一种正在出现的病原体，影响着全球人口，感染率迅速上升。CHIKV是甲病毒属的一种虫媒病毒，主要存在于热带地区，通过蚊子传播。气候变化加速了这些传播媒介在全球的传播，导致包括欧洲部分地区在内的非热带地区爆发疫情。由于缺乏抗病毒疗法以及与其他病毒共感染的可能性，CHIKV成为一个重大的公共卫生问题。CHIKV的复制依赖于nsP2半胱氨酸蛋白酶的活性，将其病毒多蛋白切割成功能性的非结构蛋白和结构蛋白。靶向nsP2蛋白酶是抗病毒疗法开发的一个有前景的策略。在这项研究中，利用噬菌体展示技术从肽库中筛选出目标蛋白酶的潜在结合物。对鉴定出的肽进行生物物理和生化分析，评估它们的抑制潜力。在鉴定出的六种肽（命名为P1 - P6）中，有四种对nsP2蛋白酶（nsP2）表现出抑制作用。肽P1表现出最强的抑制作用，半数抑制浓度（IC）为4.6±1.9μM，且细胞毒性较低。通过圆二色光谱法进行的二级结构分析和同源建模表明，P1采用α - 螺旋构象。最后，分子动力学模拟使我们能够研究nsP2活性位点的动力学，并利用分子对接预测P1的正构结合模式，从而深入了解蛋白质 - 肽相互作用。这些发现强调了肽P1作为一种先导化合物在基孔肯雅病毒研究背景下进行进一步研究的潜力。
