da Rocha João Augusto Pereira, da Costa Renato Araújo, da Rocha Elaine Cristina Medeiros, Machado Alencar Kolinski, Bick Djenifer Leticia Ulrich, Fagan Solange Binotto, Wanzeller Ana Lucia Monteiro, Gomes Micael Douglas de Souza, Rodrigues João Lucas Lima, do Rego José de Arimatéia Rodrigues, Brasil Davi do Socorro Barros, Lima Anderson H
Laboratory of Modeling and Computational Chemistry, Federal Institute of Education, Science and Technology of Pará (IFPA) Campus Bragança, Bragança 68600-000, PA, Brazil.
Graduate Program in Chemistry, Federal University of Pará (UFPA), Belém 66075-110, PA, Brazil.
ACS Omega. 2025 Aug 6;10(32):35865-35877. doi: 10.1021/acsomega.5c02814. eCollection 2025 Aug 19.
Chikungunya virus (CHIKV) is an emerging arbovirus that causes acute and chronic disease with significant public health concerns. Although recent vaccines have been licensed, no specific antiviral treatments are currently available. Targeting viral entry remains a promising strategy, particularly by inhibiting the E1-E2 glycoprotein complex, which mediates host cell attachment and membrane fusion. Natural products such as piperine have demonstrated broad-spectrum bioactivity, including antiviral properties, making them attractive candidates for drug discovery. This study employed a multidisciplinary approach, combining assays and computational modeling to evaluate the antiviral potential of piperine against CHIKV. Cytotoxicity assays were conducted in VERO cells, followed by plaque reduction assays to assess piperine's effects at different stages of CHIKV infection. In parallel, molecular docking, MD simulations, and MM/GBSA calculations revealed a stable and energetically favorable binding of piperine to the E1-E2 fusion pocket. The results demonstrated that piperine exerts a time-dependent antiviral effect, with the most pronounced inhibition observed when administered during or after infection, suggesting activity beyond the initial entry step. Docking studies revealed that piperine binds within the E1-E2 fusion pocket, forming stable interactions with key residues such as MET88, LEU16, and TYR15, which are functionally important for CHIKV fusion. MD simulations confirmed that piperine maintains stable interactions at this interface and subtly alters the conformational dynamics of the glycoprotein complex. Cytotoxicity analysis showed that piperine is safe at low concentrations (0.001-10 μg/mL), while higher doses (≥100 μg/mL) induced oxidative stress and DNA damage in a dose-dependent manner. Collectively, these findings highlight piperine as a promising antiviral candidate targeting the CHIKV E1-E2 glycoprotein complex. Future studies should focus on structure-based optimization, pharmacokinetics, and efficacy to support the development of piperine-based antiviral therapies.
基孔肯雅病毒（CHIKV）是一种新兴的虫媒病毒，可引发急性和慢性疾病，引起了重大的公共卫生关注。尽管近期已有疫苗获批上市，但目前尚无特异性抗病毒治疗方法。靶向病毒进入过程仍然是一种有前景的策略，特别是通过抑制介导宿主细胞附着和膜融合的E1-E2糖蛋白复合物。胡椒碱等天然产物已显示出广谱生物活性，包括抗病毒特性，使其成为药物研发的有吸引力的候选物。本研究采用多学科方法，结合实验分析和计算建模来评估胡椒碱对CHIKV的抗病毒潜力。在VERO细胞中进行细胞毒性分析，随后进行蚀斑减少分析，以评估胡椒碱在CHIKV感染不同阶段的作用。同时，分子对接、分子动力学模拟和MM/GBSA计算显示胡椒碱与E1-E2融合口袋形成稳定且能量有利的结合。结果表明，胡椒碱发挥时间依赖性抗病毒作用，在感染期间或感染后给药时观察到最明显的抑制作用，提示其活性超出了初始进入步骤。对接研究表明，胡椒碱结合在E1-E2融合口袋内，与MET88、LEU16和TYR15等关键残基形成稳定相互作用，这些残基对CHIKV融合具有重要功能。分子动力学模拟证实，胡椒碱在该界面维持稳定相互作用，并微妙地改变糖蛋白复合物的构象动力学。细胞毒性分析表明，胡椒碱在低浓度（0.001-10μg/mL）下是安全的，而较高剂量（≥100μg/mL）以剂量依赖性方式诱导氧化应激和DNA损伤。总体而言，这些发现突出了胡椒碱作为靶向CHIKV E1-E2糖蛋白复合物的有前景的抗病毒候选物。未来的研究应集中在基于结构的优化、药代动力学和疗效方面，以支持基于胡椒碱的抗病毒疗法的开发。
