β-L-2',3'-双脱氧-5-氟胞苷在体外对乙型肝炎病毒复制的抑制作用

Inhibition of the Replication of Hepatitis B Virus in Vitro by β-L-2',3'-Dideoxy- 5-Fluoro-Cytidine

研究β-L－2'，3'-双脱氧-5-氟胞苷的抗乙型肝炎病毒作用。方法：从2．2．15细胞培养液中提取乙肝病毒DNA，用DNA印迹法分析药物对乙肝病毒DNA复制的影响。观察药物对人T-成淋巴样细胞的生长抑制作用，用狭线印迹法检测人T-成淋巴样细胞的线粒体DNA。结果：β-L-2'，3'-双脱氧-5-氟胞苷对乙肝病毒DNA合成的半数抑制浓度为0．05μmol／L，但其抗乙肝病毒作用是可逆的。对人T-成淋巴样细胞的半数生长抑制浓度为67μmol／L。选择指数远高于2'，3'-双脱氧胞苷。在浓度高到足以抑制乙肝病毒复制达90％以上时，对细胞线粒体DNA合成并无影响。结论：β-L-2'，3'-双脱氧-5-氟胞苷在体外有较强的选择性抗乙肝病毒作用。

Objective: To study the antiviral activity against hepatitis B virus(HBV) of β-L-2', 3'-dideoxy-5-fluoro-cytidine. Methods:HBV DNA was extracted from 2. 2. 15 cell cultures. Southern blot analysis of the DNAwas performed to determine the effect on HBV DNA replication. A growth inhibition assay was used to ascertainthe cytotoxicity to human T-lymphoblastoid(CEM) cells. Mitochondrial DNA (mtDNA) content in CEM cellswas determined by slot-blotting. Results:The 50% inhibition concentration against HBV DNA synthesis of β-L-2', 3'-dideoxy-5-fluoro-cytidine was 0. 05 μmol/L. Its antiviral action was reversible. The concentration of β-L-2', 3'-dideoxy-5-fluoro-cytidine required to inhibit 50% CEM cell growth in culture was 67 μmol/L. The selective index of β-L-2', 3'-dideoxy-5-fluoro-cytidine was much higher than that of 2', 3'-dideoxycytidine. β-L-2', 3'- dideoxy-5-fluoro-cytidine did not inhibit mtDNA synthesis in CEM cells at concentrations that caused over 90%inhibition of HBV DNA replication. Conclusion: β-L-2', 3'-dideoxy-5-fluoro-cytidine was shown to have potentand selective antiviral activity against HBV in vitro.