Conformational changes of viral glycoproteins govern the fusion of viral and cellular membranes in the entry of enveloped viruses. Peptides mimicking domains of viral glycoproteins are apt to interfere with the fusion event, likely hampering the conformational rearrangements from the pre- to the post-fusion structures. We previously developed a peptide sequence with a high potential to inhibit the entry of herpes simplex type 1, which was able to trap glycoprotein B at an intermediate stage, arresting fusion. We propose that similarly to other viruses, membrane targeting through cholesterol conjugation may potently block fusion. The peptide conjugated to polyethylenglycol and cholesterol interacts with viral and cell membranes thanks to the presence of cholesterol and blocks the conformational rearrangements of the glycoprotein B. Here, we also probed the effect of the linker (polyethylenglycol) length on the activity. By targeting the peptide gBh1m to the membranes where fusion occurs and by engineering sequences with increased binding affinity for gB we have enhanced the antiviral potency of our prototype inhibitors. Our results provide proof of concept for the application of cholesterol tagging to develop inhibitors of HSV-1.

病毒糖蛋白的构象变化决定了包膜病毒进入时病毒膜和细胞膜的融合。模仿病毒糖蛋白结构域的肽易于干扰融合事件，可能会阻碍融合前结构到融合后结构的构象重排。我们先前开发了一种具有高潜力的肽序列，可以抑制1型单纯疱疹的进入，这种肽序列能够在中间阶段捕获糖蛋白B，从而阻止融合。我们提出，与其他病毒类似，通过胆固醇结合作用靶向膜可能会有效地阻止融合。由于胆固醇的存在，与聚乙二醇和胆固醇偶联的肽与病毒和细胞膜相互作用，并阻止糖蛋白B的构象重排。我们还探讨了接头（聚乙二醇）长度对活性的影响。通过将肽gBh1m靶向到发生融合的膜上，并通过对gB具有增加的结合亲和力的工程序列，我们增强了原型抑制剂的抗病毒效力。我们的结果为应用胆固醇标签开发HSV-1抑制剂提供了概念验证。