Fu L-L, Liu J, Chen Y, Wang F-T, Wen X, Liu H-Q, Wang M-Y, Ouyang L, Huang J, Bao J-K, Wei Y-Q
College of Life Sciences, Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, Sichuan University, Chengdu, 610064, China.
Cell Prolif. 2014 Aug;47(4):326-35. doi: 10.1111/cpr.12117. Epub 2014 Jun 26.
The aim of this study was to explore sodium taurocholate co-transporting polypeptide (NTCP) exerting its function with hepatitis B virus (HBV) and its targeted candidate compounds, in HBV therapy.
Identification of NTCP as a novel HBV target for screening candidate small molecules, was used by phylogenetic analysis, network construction, molecular modelling, molecular docking and molecular dynamics (MD) simulation. In vitro virological examination, q-PCR, western blotting and cytotoxicity studies were used for validating efficacy of the candidate compound.
We used the phylogenetic analysis of NTCP and constructed its protein-protein network. Also, we screened compounds from Drugbank and ZINC, among which five were validated for their authentication in HepG 2.2.15 cells. Then, we selected compound N4 (azelastine hydrochloride) as the most potent of them. This showed good inhibitory activity against HBsAg (IC50 = 7.5 μm) and HBeAg (IC50 = 3.7 μm), as well as high SI value (SI = 4.68). Further MD simulation results supported good interaction between compound N4 and NTCP.
In silico analysis and experimental validation together demonstrated that compound N4 can target NTCP in HepG2.2.15 cells, which may shed light on exploring it as a potential anti-HBV drug.
本研究旨在探索牛磺胆酸钠共转运多肽（NTCP）在乙型肝炎病毒（HBV）治疗中发挥其功能的作用机制及其靶向候选化合物。
通过系统发育分析、网络构建、分子建模、分子对接和分子动力学（MD）模拟，鉴定NTCP作为筛选候选小分子的新型HBV靶点。体外病毒学检测、q-PCR、蛋白质印迹和细胞毒性研究用于验证候选化合物的疗效。
我们对NTCP进行了系统发育分析并构建了其蛋白质-蛋白质网络。此外，我们从Drugbank和ZINC中筛选化合物，其中5种在HepG 2.2.15细胞中进行了验证。然后，我们选择化合物N4（盐酸氮卓斯汀）作为其中最有效的一种。它对HBsAg显示出良好的抑制活性（IC50 = 7.5μm）和对HBeAg的抑制活性（IC50 = 3.7μm），以及高SI值（SI = 4.68）。进一步的MD模拟结果支持化合物N4与NTCP之间有良好的相互作用。
计算机模拟分析和实验验证共同表明，化合物N4可以在HepG2.2.15细胞中靶向NTCP，这可能为将其开发为潜在的抗HBV药物提供思路。
