Charité-Universitätsmedizin Berlin, Institute of Virology, Berlin, Germany.
Universitätsklinikum Erlangen, Institute of Clinical and Molecular Virology, Erlangen, Germany.
Antimicrob Agents Chemother. 2021 Mar 18;65(4). doi: 10.1128/AAC.02103-20.
Herpesviruses are widespread and can cause serious illness. Many currently available antiviral drugs have limited effects, result in rapid development of resistance, and often exhibit dose-dependent toxicity. Especially for human cytomegalovirus (HCMV), new well-tolerated compounds with novel mechanisms of action are urgently needed. In this study, we characterized the antiviral activity of two new diazadispiroalkane derivatives, 11826091 and 11826236. These two small molecules exhibited strong activity against low-passage-number HCMV. Pretreatment of cell-free virus with these compounds greatly reduced infection. Time-of-addition assays where 11826091 or 11826236 was added to cells before infection, before and during infection, or during or after infection demonstrated an inhibitory effect on early steps of infection. Interestingly, 11826236 had an effect by addition to cells after infection. Results from entry assays showed the major effect to be on attachment. Only 11826236 had a minimal effect on penetration comparable to heparin. Further, no effect on virus infection was found for cell lines with a defect in heparan sulfate expression or lacking all surface glycosaminoglycans, indicating that these small molecules bind to heparan sulfate on the cell surface. To test this further, we extended our analyses to pseudorabies virus (PrV), a member of the , which is known to use cell surface heparan sulfate for initial attachment via nonessential glycoprotein C (gC). While infection with PrV wild type was strongly impaired by 11826091 or 11826236, as with heparin, a mutant lacking gC was unaffected by either treatment, demonstrating that primary attachment to heparan sulfate via gC is targeted by these small molecules.
疱疹病毒广泛存在，可导致严重疾病。目前许多抗病毒药物的效果有限，导致耐药性迅速发展，且常表现出剂量依赖性毒性。特别是对于人巨细胞病毒（HCMV），急需具有新型作用机制的新的耐受性良好的化合物。在本研究中，我们对两种新型氮杂二螺环烷烃衍生物 11826091 和 11826236 的抗病毒活性进行了表征。这两种小分子对低传代数 HCMV 具有很强的活性。用这些化合物预处理无细胞病毒可大大降低感染性。在添加实验中，将 11826091 或 11826236 在感染前、感染时和感染后加入细胞，结果表明这些化合物对感染的早期步骤具有抑制作用。有趣的是，11826236 在感染后加入细胞也具有抑制作用。进入实验的结果表明，主要作用是在附着阶段。只有 11826236 对穿透有最小的影响，与肝素相当。此外，在缺乏硫酸乙酰肝素表达或缺乏所有表面糖胺聚糖的细胞系中，没有发现这些小分子对病毒感染有影响，表明这些小分子与细胞表面的硫酸乙酰肝素结合。为了进一步验证这一点，我们将分析扩展到伪狂犬病病毒（PrV），它是疱疹病毒科的一员，已知其通过非必需糖蛋白 C（gC）利用细胞表面硫酸乙酰肝素进行初始附着。虽然 11826091 或 11826236 强烈抑制了 PrV 野生型的感染，但与肝素一样，缺乏 gC 的突变体不受两种处理的影响，表明这些小分子通过 gC 与硫酸乙酰肝素的初始附着是它们的作用靶点。
