IN VITRO INHIBITION OF HEPATITIS B VIRUS BY 2',3'-DIDEOXYNUCLEOSIDES

HepG2来源的肝母细胞瘤细胞(2.2.15) 在存在或不存在的情况下培养乙肝病毒(HBV) 抗逆转录病毒药物2‘，3’-二脱氧鸟苷(DDG)，2‘，3’-二脱氧肌苷 (DDI)，或3‘-叠氮-2’，3‘-二脱氧胸苷(AZT)。所有这些特工 抑制2.2.15细胞中的病毒复制 在不影响细胞生长的情况下抑制染色体外的HBVDNA。在这些人中 三个双脱氧核苷，DDG是最有效的试剂，减少病毒 复制速度高达97%。Northern印迹分析显示没有明显的 处理细胞和未处理细胞前基因组RNA的差异 提示双脱氧核苷可抑制乙肝病毒的复制。 逆转录阶段。不同给药时间的效果观察 暴露表明，这些制剂可以抑制乙肝病毒复制，即使在 在培养后期添加，表明如果由 逆转录酶被持续抑制，染色体外DNA 分子是不稳定的，在一段时间后就会消失。高效液相色谱仪 用~(3)H-DDG分析表明，DDG的合成磷酸化为AN 假定的活性代谢物ddGTP存在于2.2.15细胞中。 这些数据表明，2‘，3’-双脱氧核苷可能是潜在的 靶向治疗乙肝病毒感染的实验药物 逆转录步骤，尽管数据没有涉及体内 毒性概况或治疗指数。

HepG2-derived hepatoblastoma cells (2.2.15) which actively produce hepatitis B virus (HBV) were cultured in the presence or absence of the antiretroviral agents 2',3'-dideoxyguanosine (ddG), 2',3'-dideoxyinosine (ddI), or 3'-azido-2',3'-dideoxythymidine (AZT). All of these agents diminished the viral replication in 2.2.15 cells as assessed by the amount of extrachromosomal HBV DNA without affecting cellular growth. Among the three dideoxynucleosides, ddG was the most potent agent, diminishing viral replication by as much as 97%. Northern blot analysis revealed no apparent difference between treated and untreated cells in the pregenomic RNA profile, suggesting that dideoxynucleosides suppress HBV replication at the stage of reverse transcription. The effect of varying the time of drug exposure showed that these agents can suppress HBV replication even when added late in culture, suggesting that if de novo DNA synthesis mediated by reverse transcriptase is continuously suppressed, the extrachromosomal DNA molecules are unstable and disappear after a certain period of time. HPLC analyses using (3)H-ddG showed that anabolic phosphorylation of ddG to an assumed active metabolite, ddGTP, occurs in 2.2.15 cells. These data suggest that 2',3'-dideoxynucleosides may be potential experimental drugs for the treatment of HBV infection by targeting the reverse transcription step, although the data do not address the in vivo toxicity profile or therapeutic index.