Capsid Assembly Modulators Have a Dual Mechanism of Action in Primary Human Hepatocytes Infected with Hepatitis B Virus
Capsid Assembly Modulators Have a Dual Mechanism of Action in Primary Human Hepatocytes Infected with Hepatitis B Virus
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DOI：
10.1128/aac.00560-17
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发表时间：
2017-08-01
影响因子：
4.9
通讯作者：
Pauwels, Frederik
中科院分区：
文献类型：
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作者：
Berke, Jan Martin;Dehertogh, Pascale;Pauwels, Frederik
乙肝病毒衣壳的组装是病毒繁殖的关键步骤，由核心蛋白介导。由于其在病毒生命周期中的多种功能，CORE成为新的抗病毒治疗的有吸引力的靶点。衣壳组装调节剂(CAM)加速衣壳组装的动力学，阻止聚合酶-前基因组RNA(Pol-pgRNA)复合体的包裹，从而阻止病毒复制。CaM JNJ-632是一种新的、有效的体外复制乙肝病毒的抑制剂，通过大小排斥层析和电子显微镜研究表明，它可以诱导形态完整的病毒衣壳的形成。在原代人肝细胞中的抗病毒图谱显示，当化合物与病毒接种剂一起加入时，CAMS以剂量依赖的方式阻止共价闭合环状DNA的形成，而核(T)类似物(NAS)不能。这种保护作用表现为细胞内HBVRNA水平的剂量依赖性降低，以及细胞培养上清液中HBe/CAg和HBs Ag水平的降低。对另一种CAM(BAY41-4109)也进行了同样的观察，表明机械效应而不是化合物特有的效应起到了作用。我们的数据表明，CAM具有双重作用机制，抑制病毒生命周期的早期和晚期。这些效应明显地将CAMS与NAS区分开来，当单独或与目前的护理标准结合使用时，可能会在临床环境中转化为更高的功能治愈率。
Hepatitis B virus (HBV) capsid assembly is a critical step in the propagation of the virus and is mediated by the core protein. Due to its multiple functions in the viral life cycle, core became an attractive target for new antiviral therapies. Capsid assembly modulators (CAMs) accelerate the kinetics of capsid assembly and prevent encapsidation of the polymerase-pregenomic RNA (Pol-pgRNA) complex, thereby blocking viral replication. CAM JNJ-632 is a novel and potent inhibitor of HBV replication in vitro across genotypes A to D. It induces the formation of morphologically intact viral capsids, as demonstrated by size exclusion chromatography and electron microscopy studies. Antiviral profiling in primary human hepatocytes revealed that CAMs prevented formation of covalently closed circular DNA in a dose-dependent fashion when the compound was added together with the viral inoculum, whereas nucleos(t) ide analogues (NAs) did not. This protective effect translated into a dose-dependent reduction of intracellular HBV RNA levels as well as reduced HBe/cAg and HBsAg levels in the cell culture supernatant. The same observation was made with another CAM (BAY41-4109), suggesting that mechanistic rather than compound-specific effects play a role. Our data show that CAMs have a dual mechanism of action, inhibiting early and late steps of the viral life cycle. These effects clearly differentiate CAMs from NAs and may translate into higher functional cure rates in a clinical setting when given alone or in combination with the current standard of care.