Zhang Han, Shi Shuxin, Sun Lujia, Li Shuangqu, Zhang Yan, Li Ziyue, Hou Jingjing, Li Pingan, Shen Jingshan, Cheng Xi, Jiang Shibo, Gao Zhaobing, Wang Xinling, Jiang Xiangrui, Xia Bingqing
School of Chinese Materia Medica, Nanjing University of Chinese Medicine, Nanjing 210046, PR China.
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, PR China.
Curr Res Microb Sci. 2025 May 22;9:100409. doi: 10.1016/j.crmicr.2025.100409. eCollection 2025.
The SARS-CoV-2-encoded 2-E channel is critical in the viral life cycle and pathogenesis. By facilitating viral replication, it promotes the dysregulation of inflammatory pathways, leading to cytokine storm, and triggers DNA damage response (DDR), thus exacerbating disease progression. The 2-E channel, a viroporin, is a promising antiviral target. However, the lack of specific inhibitors and effective screening methods has hindered therapeutic exploitation of the 2-E channel. To address this gap, we report on a fluorescence-based screening assay that targets the 2-E channel activity, resulting in the identification of potential inhibitory molecules. After performing both electrophysiological studies and surface plasmon resonance (SPR) analyses, we identified the top-ranked candidate, TPN10518, as a pore-blocking inhibitor of the 2-E channel. TPN10518 binds to a hydrophobic pocket in the C-terminal vestibule of the 2-E channel, thereby inhibiting its activity. Functional evaluation showed that TPN10518 exhibits significant antiviral efficacy , while, at the same time, effectively protecting against 2-E channel-mediated host damage and suppressing cytokine storm caused by dysregulated homeostasis of inflammatory pathways . Therefore, our work introduces a screening method for targeting 2-E channels, establishes the 2-E channel as a viable therapeutic target against SARS-CoV-2, and identifies TPN10518 as a promising antiviral candidate.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）编码的2-E通道在病毒生命周期和发病机制中至关重要。通过促进病毒复制，它会导致炎症途径失调，引发细胞因子风暴，并触发DNA损伤反应（DDR），从而加剧疾病进展。2-E通道作为一种病毒离子通道蛋白，是一个很有前景的抗病毒靶点。然而，缺乏特异性抑制剂和有效的筛选方法阻碍了对2-E通道的治疗开发。为了填补这一空白，我们报道了一种基于荧光的筛选试验，该试验针对2-E通道活性，从而鉴定出潜在的抑制分子。在进行电生理研究和表面等离子体共振（SPR）分析后，我们确定排名第一的候选物TPN10518是2-E通道的孔道阻断抑制剂。TPN10518与2-E通道C端前庭的一个疏水口袋结合，从而抑制其活性。功能评估表明，TPN10518具有显著的抗病毒功效，同时能有效防止2-E通道介导的宿主损伤，并抑制由炎症途径稳态失调引起的细胞因子风暴。因此，我们的工作引入了一种针对2-E通道的筛选方法，确立了2-E通道作为抗SARS-CoV-2的可行治疗靶点，并确定TPN10518为一种有前景的抗病毒候选物。
