Clofazimine targeting the spike protein and RdRp exhibits highly efficient antiviral activity against porcine epidemic diarrhea virus <i>in vitro</i>

Shuting Zhou; Junrui Zhu; Houde Zhao; Zixin Huang; Kangqi Zheng; Fan Xia; Yufan Xu; Guocheng Zhao; Jijie Jiang; En Zhang; Haoyang Nian; Li Cui; Tao Sun; Xiangfeng Wang; Yanjun Zhou; Zhibiao Yang; Zhe Wang

doi:10.1016/j.virs.2025.05.012

June 2025

. doi: 10.1016/j.virs.2025.05.012

Citation: Shuting Zhou, Junrui Zhu, Houde Zhao, Zixin Huang, Kangqi Zheng, Fan Xia, Yufan Xu, Guocheng Zhao, Jijie Jiang, En Zhang, Haoyang Nian, Li Cui, Tao Sun, Xiangfeng Wang, Yanjun Zhou, Zhibiao Yang, Zhe Wang. Clofazimine targeting the spike protein and RdRp exhibits highly efficient antiviral activity against porcine epidemic diarrhea virus in vitro .VIROLOGICA SINICA, 2025, 40(3) : 477-490. http://dx.doi.org/10.1016/j.virs.2025.05.012

氯法齐明通过靶向刺突蛋白和RdRp在体外对猪流行性腹泻病毒表现出高效的抗病毒活性

周书亭 ^a,b ,
朱钧锐 ^c ,
赵厚德 ^a,b ,
黄梓鑫 ^a,b ,
郑康琪 ^a,b ,
夏凡 ^d ,
徐宇帆 ^a,b ,
赵国成 ^a,b ,
蒋霁捷 ^a,b ,
张恩 ^b ,
念皞洋 ^b ,
崔立 ^b ,
孙涛 ^b ,
王向峰 ^d ,
周艳君 ^c ,
杨志彪 ^a,b,, ,
王哲 ^a,b,,

a 上海交通大学农业与生物学院/上海市现代种业协同创新中心;
b 上海交通大学农业与生物学院/上海市兽医生物技术重点实验室;
c 中国农业科学院上海兽医研究所猪传染病研究室;
d 中国农业大学农学与生物技术学院

通讯作者： 杨志彪, zbyang@sjtu.edu.cn; 王哲, wangz@sjtu.edu.cn
收稿日期： 2024-09-14
录用日期： 2025-05-28

摘要

猪流行性腹泻病毒（PEDV）感染会导致新生仔猪急性水样腹泻，给养猪业造成重大经济损失。本研究表明，氯法齐明（CFZ）在体外以剂量依赖性方式显著抑制PEDV复制，且细胞毒性可忽略不计。通过时间添加实验，我们发现CFZ能有效破坏病毒感染周期的多个阶段。利用CoV-RdRp-Gluc报告系统，我们评估了CFZ对PEDV RNA依赖性RNA聚合酶（RdRp）的抑制作用，并测得其IC50值为0.1364 μM，表明其高效性。分子对接研究进一步证实，CFZ在PEDV的刺突蛋白和RdRp蛋白活性位点具有高结合亲和力。对VeroE6细胞在有无CFZ处理下的转录组分析显示，感染后8小时（hpi）转录活性发生显著变化。此外，CFZ与核苷类似物联用可增强其在体外的抗PEDV效果。本研究强调了CFZ作为抗PEDV潜在治疗药物的可行性。
- 猪流行性腹泻病毒（PEDV）
- , 氯法齐明（CFZ）
- , 刺突蛋白
- , RdRp
- , 灭活

Clofazimine targeting the spike protein and RdRp exhibits highly efficient antiviral activity against porcine epidemic diarrhea virus in vitro

Shuting Zhou ^a,b ,
Junrui Zhu ^c ,
Houde Zhao ^a,b ,
Zixin Huang ^a,b ,
Kangqi Zheng ^a,b ,
Fan Xia ^d ,
Yufan Xu ^a,b ,
Guocheng Zhao ^a,b ,
Jijie Jiang ^a,b ,
En Zhang ^b ,
Haoyang Nian ^b ,
Li Cui ^b ,
Tao Sun ^b ,
Xiangfeng Wang ^d ,
Yanjun Zhou ^c ,
Zhibiao Yang ^a,b,, ,
Zhe Wang ^a,b,,

a. Shanghai Collaborative Innovation Center of Agri-Seeds/School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China;
b. Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China;
c. Department of Swine Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, China;
d. College of Agronomy and Biotechnology, China Agricultural University, Beijing, 100094, China

Corresponding author: Zhibiao Yang, zbyang@sjtu.edu.cn Zhe Wang, wangz@sjtu.edu.cn
Received Date: 14 September 2024
Accepted Date: 28 May 2025

Abstract

Porcine epidemic diarrhea virus (PEDV) infection causes acute watery diarrhea in neonatal piglets, leading to substantial economic losses within the pig farming industry. This study demonstrates that clofazimine (CFZ) significantly inhibits PEDV replication in a dose-dependent manner in vitro, with negligible cytotoxicity. Findings from our time-of-addition assays indicate that CFZ effectively disrupts multiple stages of the viral infection cycle. Using a CoV-RdRp-Gluc reporter system, we evaluated the potency of CFZ against PEDV RNA-dependent RNA polymerase (RdRp), and determined a low IC₅₀ value of 0.1364 μM. Molecular docking studies further confirmed that CFZ has high binding affinity at the active sites of the spike protein and RdRp protein in PEDV. Transcriptome analysis of Vero E6 cells, with and without CFZ treatment, revealed a significant change in transcriptional activity at 8 h post-infection (hpi). Moreover, the simultaneous application of CFZ and nucleoside analogs showed enhanced the anti-PEDV effect of CFZ in vitro. Our study underscores the potential of CFZ as a viable therapeutic agent against PEDV.
- Porcine epidemic diarrhea virus (PEDV)
- , Clofazimine (CFZ)
- , Spike
- , RdRp
- , Inactivation

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