Novel Hepatitis B Virus Capsid-Targeting Antiviral That Aggregates Core Particles and Inhibits Nuclear Entry of Viral Cores
Novel Hepatitis B Virus Capsid-Targeting Antiviral That Aggregates Core Particles and Inhibits Nuclear Entry of Viral Cores
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DOI：
10.1021/acsinfecdis.8b00235
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发表时间：
2019-05-01
影响因子：
5.3
通讯作者：
Sarafianos, Stefan G.
中科院分区：
文献类型：
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作者：
Huber, Andrew D.;Pineda, Dallas L.;Sarafianos, Stefan G.
估计有2.4亿人慢性感染B型肝炎病毒（HBV），可导致肝病、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能治愈。因此，具有新的作用机制的新治疗是HBV治疗的高度期望。病毒核心蛋白（Cp）由于其在HBV生命周期中的重要作用而作为可能的治疗靶点受到关注。已经详细描述了几类衣壳组装效应物（CAEs），并且这些化合物都通过不同的机制增加衣壳组装速率但抑制HBV复制。在这项研究中，我们开发了一种基于热位移的筛选方法，用于CAE发现和表征，填补了captain靶向分子高通量筛选方法中急需的空白。使用这种方法，然后进行基于细胞的筛选，我们将化合物HF 9 C6鉴定为对HBV复制具有低微摩尔效力的CAE。HF 9 C6引起大的多衣壳聚集体时，衣壳在体外组装，并通过透射电子显微镜分析。有趣的是，当用HF 9 C6处理HBV表达细胞时，Cp被排除在细胞核之外，这表明该化合物可以抑制Cp和衣壳进入细胞核。此外，Cp的突变扫描表明HF 9 C6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作用机制中具有作用。
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 multicapsid 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 the CAE mechanism of action.