Sun Senfeng, Sun Tingting, Lei Xinyu, Li Fangfei, Wang Jing, Zhou Rui, Li Quanjie, Yi Dongrong, Wang Jing, Jin Hongwei, Cen Shan, Chen Shizhong
School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical School, Beijing 100050, China.
Phytomedicine. 2025 Oct;146:157105. doi: 10.1016/j.phymed.2025.157105. Epub 2025 Aug 5.
Zika virus (ZIKV) belonging to the Flaviviridae family causes critical neurological abnormalities, including congenital microcephaly and Guillain-Barré syndrome. Despite being a major public health concern, effective therapeutic interventions against ZIKV remain unavailable. NS2B-NS3 (ZIKVpro), the viral serine protease critical for polyprotein processing as well as viral replication, is a potential target for developing antiviral agents PURPOSE: This study aims to develop ZIKV inhibitors targeting the ZIKVpro from natural products.
Surface plasmon resonance (SPR) based assay were used for drug screening from a natural product library. qRT-PCR, Western blot and plaque assay was used for the evaluation of their antiviral activity and studies of their mechanisms of action. Hydrogen-deuterium exchange mass spectrometry (HDXMS) analysis and Molecular dynamics (MD) simulation were used for confirmation of action site.
We demonstrate that neferine, a bisbenzylisoquinoline alkaloid derived from the green embryo of the mature seeds of Nelumbo nucifera Gaertn (Plumula Nelumbinis), exhibits potent inhibitory effects against ZIKV replication in vitro and in vivo. Mechanistic analyses revealed that neferine directly binds to ZIKVpro, interacting with catalytic residues Tyr161 and Ser135, thereby suppressing protease activity in enzymatic assays and impairing polyprotein processing in infected cells. Notably, neferine also demonstrated broad-spectrum antiviral activity against Dengue virus, suggesting its potential as a pan-Flavivirus inhibitor.
Our results highlight neferine's therapeutic potential for combating ZIKV and related Flavivirus infections, providing a robust foundation for further drug development.
寨卡病毒（ZIKV）属于黄病毒科，可导致严重的神经异常，包括先天性小头畸形和吉兰-巴雷综合征。尽管它是一个重大的公共卫生问题，但针对寨卡病毒的有效治疗干预措施仍然缺乏。NS2B-NS3（ZIKVpro）是病毒丝氨酸蛋白酶，对多蛋白加工以及病毒复制至关重要，是开发抗病毒药物的潜在靶点。目的：本研究旨在从天然产物中开发靶向ZIKVpro的寨卡病毒抑制剂。
基于表面等离子体共振（SPR）的分析方法用于从天然产物库中进行药物筛选。qRT-PCR、蛋白质免疫印迹和蚀斑试验用于评估其抗病毒活性并研究其作用机制。氢-氘交换质谱（HDXMS）分析和分子动力学（MD）模拟用于确认作用位点。
我们证明，荷叶碱，一种从莲（Nelumbo nucifera Gaertn）成熟种子的绿色胚中提取的双苄基异喹啉生物碱，在体外和体内均对寨卡病毒复制表现出强大的抑制作用。机制分析表明，荷叶碱直接与ZIKVpro结合，与催化残基Tyr161和Ser135相互作用，从而在酶促试验中抑制蛋白酶活性，并损害感染细胞中的多蛋白加工。值得注意的是，荷叶碱还对登革病毒表现出广谱抗病毒活性，表明其作为泛黄病毒抑制剂的潜力。
我们的结果突出了荷叶碱在对抗寨卡病毒和相关黄病毒感染方面的治疗潜力，为进一步的药物开发提供了坚实的基础。
