新型乙型肝炎病毒衣壳靶向抗病毒，聚集核心颗粒和抑制病

StephenW

ACS Infect Dis. 2018 Dec 24. doi: 10.1021/acsinfecdis.8b00235. [Epub ahead of print]
Novel Hepatitis B Virus Capsid-Targeting Antiviral that Aggregates Core Particles and Inhibits Nuclear Entry of Viral Cores.
Huber AD, Pineda DL, Liu D, Boschert KN, Gres AT, Wolf JJ, Coonrod EM, Tang J, Laughlin TG, Yang Q, Puray-Chavez M, Ji J, Singh K, Kirby KA, Wang Z, Sarafianos SG.
Abstract

An estimated 240 million are chronically infected with hepatitis B virus (HBV), which can lead to liver disease, cirrhosis, and hepatocellular carcinoma. Currently, HBV treatment options include only nucleoside reverse transcriptase inhibitors and the immunomodulatory agent interferon alpha, and these treatments are generally not curative. New treatments with novel mechanisms of action, therefore, are highly desired for HBV therapy. The viral core protein (Cp) has gained attention as a possible therapeutic target because of its vital roles in the HBV life cycle. Several classes of capsid assembly effectors (CAEs) have been described in detail, and these compounds all increase capsid assembly rate, but inhibit HBV replication by different mechanisms. In this study, we have developed a thermal shift-based screening method for CAE discovery and characterization, filling a much-needed gap in high-throughput screening methods for capsid-targeting molecules. Using this approach followed by cell-based screening, we identified the compound HF9C6 as a CAE with low micromolar potency against HBV replication. HF9C6 caused large multi-capsid aggregates when capsids were assembled in vitro and analyzed by transmission electron microscopy. Interestingly, when HBV-expressing cells were treated with HF9C6, Cp was excluded from cell nuclei, suggesting that this compound may inhibit nuclear entry of Cp and capsids. Furthermore, mutational scanning of Cp suggested that HF9C6 binds the known CAE binding pocket, indicating that key Cp-compound interactions within this pocket have a role in determining CAE mechanism of action.

PMID:
    30582687
DOI:
    10.1021/acsinfecdis.8b00235

StephenW

ACS Infect Dis。 2018年12月24日doi：10.1021 / acsinfecdis.8b00235。 [提前打印]
新型乙型肝炎病毒衣壳靶向抗病毒，聚集核心颗粒和抑制病毒核的核进入。
Huber AD，Pineda DL，Liu D，Boschert KN，Gres AT，Wolf JJ，Coonrod EM，Tang J，Laughlin TG，Yang Q，Puray-Chavez M，Ji J，Singh K，Kirby KA，Wang Z，Sarafianos SG。
抽象

估计有2.4亿人慢性感染乙型肝炎病毒（HBV），这可能导致肝病，肝硬化和肝细胞癌。目前，HBV治疗选择仅包括核苷逆转录酶抑制剂和免疫调节剂干扰素α，并且这些治疗通常不是治愈性的。因此，具有新作用机制的新疗法对于HBV疗法是非常需要的。病毒核心蛋白（Cp）因其在HBV生命周期中的重要作用而作为可能的治疗靶点而受到关注。已经详细描述了几类衣壳组装效应物（CAE），并且这些化合物都增加衣壳组装速率，但通过不同机制抑制HBV复制。在这项研究中，我们开发了一种基于热转移的CAE发现和表征筛选方法，填补了衣壳靶向分子高通量筛选方法的急需空白。使用该方法然后进行基于细胞的筛选，我们将化合物HF9C6鉴定为具有针对HBV复制的低微摩尔效力的CAE。当衣壳在体外组装并通过透射电子显微镜分析时，HF9C6引起大的多衣壳聚集体。有趣的是，当表达HBV的细胞用HF9C6处理时，Cp被排除在细胞核之外，这表明该化合物可能抑制Cp和衣壳的核进入。此外，Cp的突变扫描表明HF9C6结合已知的CAE结合口袋，表明该口袋内的关键Cp-化合物相互作用在确定CAE作用机制中起作用。

结论：
    30582687
DOI：
    10.1021 / acsinfecdis.8b00235