Wei Shukun, Liu Chaolun, Du Lingyu, Wu Bin, Zhong Jin, Tong Yimin, Wang Shuqing, OuYang Bo
State Key Laboratory of Molecular Biology, Centre for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
Comput Struct Biotechnol J. 2022 Oct 28;20:5902-5910. doi: 10.1016/j.csbj.2022.10.035. eCollection 2022.
The hepatitis C virus (HCV) p7 viroporin protein is essential for viral assembly and release, suggesting its unrealised potential as a target for HCV interventions. Several classes of small molecules that can inhibit p7 through allosteric mechanisms have shown low efficacy. Here, we used a high throughput virtual screen to design a panel of eight novel cyclic penta-peptides (CPs) that target the p7 channel with high binding affinity. Further examination of the effects of these CPs in viral production assays indicated that CP7 exhibits the highest potency against HCV among them. Moreover, the IC efficacy of CP7 in tests of strain Jc1-S282T suggested that this cyclopeptide could also effectively inhibit a drug-resistant HCV strain. A combination of nuclear magnetic resonance (NMR) spectroscopy and molecular dynamics (MD) simulations revealed that CP7 blocking activity relies on direct binding to the p7 channel lumen at the -terminal bottleneck region. These findings thus present a promising anti-HCV cyclic penta-peptide targeting p7 viroporin, while also describing an alternative strategy for designing a new class of p7 channel blockers for strains resistant to direct-acting antiviral agents (DAA).
丙型肝炎病毒（HCV）的p7病毒孔蛋白对于病毒的组装和释放至关重要，这表明其作为HCV干预靶点的潜力尚未得到充分发挥。几类能够通过变构机制抑制p7的小分子显示出较低的疗效。在此，我们利用高通量虚拟筛选设计了一组针对p7通道具有高结合亲和力的八种新型环五肽（CPs）。在病毒产生试验中对这些CPs作用的进一步研究表明，CP7在其中对HCV表现出最高的效力。此外，CP7在Jc1-S282T毒株试验中的IC效力表明，这种环肽也能有效抑制耐药HCV毒株。核磁共振（NMR）光谱和分子动力学（MD）模拟相结合的结果表明，CP7的阻断活性依赖于在N末端瓶颈区域直接结合到p7通道内腔。因此，这些发现展示了一种有前景的靶向p7病毒孔蛋白的抗HCV环五肽，同时也描述了一种为对直接作用抗病毒药物（DAA）耐药的毒株设计新型p7通道阻滞剂的替代策略。
