乙型肝炎病毒（HBV）进入抑制剂是颇具前景的新机制治疗乙型肝炎药物。最近发现，钠离子-牛磺胆酸-协同转运蛋白（NTCP）为HBV感染肝细胞所需的特异性靶点。红树木榄是广西京族医生治疗乙型肝炎的传统药物。在前期研究中，申请人发现木榄胚轴提取物具有抑制模型鸭体内HBV复制和保护肝细胞活性，高剂量组会抑制NTCP蛋白生成。申请人发现3个富含环亚己基己腈类化合物的活性部位（Y6、H4和H8），进一步从Y6中分离到7个具有较显著抗HBV复制的环亚己基己腈类活性化合物。本项目拟在前期工作基础上，运用HPLC-DAD(UV)-MS方法从H4和H8部位定向追踪环亚己基己腈类特征活性成分，运用现代波谱技术和化学手段鉴定其结构，借助体内和体外活性模型系统评价环亚己基己腈类化合物抗HBV复制活性，并探讨其对靶点蛋白NTCP作用，获得腈类先导化合物，研究成果为海洋来源的治疗乙型肝炎新型药物提供坚实物质基础。

HBV entry inhibitors represent a new generation of antivirals for the treatment of HBV infection, by blocking the HBV entry into the target cells. Sodium taurocholate cotransporting polypeptide (NTCP) was recently indentified and recognized as a fuctional target for HBV. Jing people is the only national marine fisheries of China. Mangrove Bruguiera gymnorrhiza is Jing's treatment of hepatitis B traditional herb medicine. In our previous study, we have discovered alcohol extract of the hypocotyl of B. gymnorrhiza (AEHB) show activities on anti-duck hepatitis B virus and rich of characteristic cyclohexylideneacetonitrile derivatives. High-dosage AEHB has an obvious protective effect on the liver function and liver tissue. Compared with control group, protein expression of NTCP in High-dosage AEHB were down-regulated.AEHB were fractionated to yield three active parts (Y6, H4 and H8) which contain cyclohexylideneacetonitrile derivatives and have protective effects potent on treatment of HBV carriers. Initial searching for bioactive metabolites from Part Y6 has led to the isolation of seven cyclohexylideneacetonitrile derivatives (of which 4 compounds are new) showing protective effects potent as a HBV inhibitor in cell model system..In order to obtain highly efficient and specific anti-HBV drugs with independent intellectual property, this project plans to systematically investigate the active anti-HBV cyclohexylideneacetonitrile derivatives from the hypocotyl of B. gymnorrhiza. The contents include: 1) targeted isolating cyclohexylideneacetonitrile derivatives from the Part H4 and Part H8 by HPLC-DAD(UV)-MS and various modern chromatography techniques; 2) determining the structures of cyclohexylideneacetonitrile derivatives by MS, 1D and 2D NMR, CD, ORD and chemical methods; 3) examining the activity of the isolated compounds based on cell model system and duck hepatitis B model system of cyclohexylideneacetonitrile derivatives; 4) studying the action targets of lead compounds through Sodium taurocholate cotransporting polypeptide (NTCP) as a funtional receptor of HBV on model cell system.The current research would provide fundamentally cyclohexylideneacetonitrile derivatives as lead compound for HBV infection management.