基于乙肝病毒X蛋白与高迁移率族蛋白1互作靶点抗乙肝病毒药物的设计、生物活性评价及其作用机制研究

乙肝病毒X蛋白(HBx) 在乙肝病毒（HBV)复制生命周期中起着重要的作用，靶向该病毒蛋白是乙肝抗病毒治疗领域的研究热点，而阐明HBx调控宿主蛋白的分子机制是发展这一领域的关键。申请者前期利用体外细胞模型，HBx转基因动物及临床肝组织标本证实了HBx-HMGB1蛋白复合体在调控肝细胞自噬中的关键作用，增强的肝细胞自噬将促进HBV持续感染。在此基础上，本课题拟通过基于定量的构效关系模型利用虚拟筛选技术从化合物库中筛选抑制HBx-HMGB1互作的全新小分子化合物。探讨小分子化合物抑制剂对HBx-HMGB1蛋白互作、蛋白表达与亚细胞定位、细胞自噬及HBV复制的影响，从体外细胞实验及体内HBV感染动物模型多层次评估小分子化合物抑制剂的生物活性并深入阐明其发挥抗HBV作用的潜在分子机制。本课题的开展将有望推动基于HBx–HMGB1蛋白互作抗HBV研发新策略，为新型抗HBV药物的研发奠定实验室基础。

Hepatitis B viral X protein (HBx) plays a vital role in the life cycle of hepatitis B virus （HBV）. Targeting this viral protein potentiates a novel avenue for HBV treatments. Exploring HBx-focused protein-protein interaction (PPIs) network would result in rich resources leading to critical insights into intricate biological regulatory systems. Using cell model, HBx-transgenic mice model and clinical specimen, our previous work indicated that HBx could interact with HMGB1, upregulate its expression and promote the cytoplasmic translocation of HMGB1. HBx-HMGB1 complex thus plays a pivotal role in the regulation of hepatocyte autophagy, the enhanced autophagy in hepatocyte would facilitate the HBV replication and persistent infection. Here, we exploited the quantitative structure–activity relationship (QSAR)-based virtual screening to identify novel HBx–HMGB1 protein-protein interaction inhibitors. The potential effects of small molecular inhibitors on the HBx-HMGB1 interaction, the expression levels and cellular localization of HBx and HMGB1, cellular autophagy and HBV replication were examined. Using cell line and animal model for HBV infection, the molecular mechanisms underlying the action of the small molecular inhibitors were further explored. Besides, in vitro and in vivo evaluation of the anti-HBV activity of the compound was further performed. Hence, targeting HBx-HMGB1 interaction could be utilized as an anti-viral strategy, and our study would provide fundamental mechanism for the therapeutic discovery of anti-HBV reagents for the patients with chronic hepatitis B (CHB).