The State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China; The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan, 430070, China; National Key Laboratory of Crop Genetic Improvement, National Centre of Plant Gene Research, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China; Department of Veterinary Medicine, College of Animal Science and Technology, Zhejiang A&F University, Hangzhou, 311300, China.
The State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China; The Cooperative Innovation Center for Sustainable Pig Production, Huazhong Agricultural University, Wuhan, 430070, China.
Antiviral Res. 2020 Jan;173:104651. doi: 10.1016/j.antiviral.2019.104651. Epub 2019 Nov 18.
Emerging coronaviruses (CoVs) primarily cause severe gastroenteric or respiratory diseases in humans and animals, and no approved therapeutics are currently available. Here, A9, a receptor tyrosine kinase inhibitor (RTKI) of the tyrphostin class, is identified as a robust inhibitor of transmissible gastroenteritis virus (TGEV) infection in cell-based assays. Moreover, A9 exhibited potent antiviral activity against the replication of various CoVs, including murine hepatitis virus (MHV), porcine epidemic diarrhea virus (PEDV) and feline infectious peritonitis virus (FIPV). We further performed a comparative phosphoproteomic analysis to investigate the mechanism of action of A9 against TGEV infection in vitro. We specifically identified p38 and JNK1, which are the downstream molecules of receptor tyrosine kinases (RTKs) required for efficient TGEV replication, as A9 targets through plaque assays, qRT-PCR and Western blotting assays. p38 and JNK1 inhibitors and RNA interference further showed that the inhibitory activity of A9 against TGEV infection was mainly mediated by the p38 mitogen-activated protein kinase (MAPK) signaling pathway. All these findings indicated that the RTKI A9 directly inhibits TGEV replication and that its inhibitory activity against TGEV replication mainly occurs by targeting p38, which provides vital clues to the design of novel drugs against CoVs.
新兴冠状病毒（CoV）主要在人类和动物中引起严重的胃肠道或呼吸道疾病，目前尚无批准的治疗方法。在这里，A9 是一种酪氨酸激酶抑制剂（RTKI）类的 tyrphostin，被鉴定为细胞水平测定中传染性胃肠炎病毒（TGEV）感染的有效抑制剂。此外，A9 对多种 CoV 的复制表现出强大的抗病毒活性，包括鼠肝炎病毒（MHV）、猪流行性腹泻病毒（PEDV）和猫传染性腹膜炎病毒（FIPV）。我们进一步进行了比较磷酸化蛋白质组学分析，以研究 A9 体外抗 TGEV 感染的作用机制。我们特别鉴定了 p38 和 JNK1，它们是 TGEV 复制所必需的受体酪氨酸激酶（RTKs）的下游分子，通过噬斑测定、qRT-PCR 和 Western blot 测定证实 A9 是其靶标。p38 和 JNK1 抑制剂和 RNA 干扰进一步表明，A9 对 TGEV 感染的抑制活性主要是通过 p38 丝裂原活化蛋白激酶（MAPK）信号通路介导的。所有这些发现表明，RTKI A9 直接抑制 TGEV 复制，其对 TGEV 复制的抑制活性主要通过靶向 p38 来实现，这为针对 CoV 的新型药物设计提供了重要线索。
