INSERM, U1052, 151 Cours Albert Thomas, Lyon 69424 Cedex 03, France.
Antiviral Res. 2011 Nov;92(2):271-6. doi: 10.1016/j.antiviral.2011.08.012. Epub 2011 Aug 18.
Long-term treatment of chronic hepatitis B with nucleos(t)ide analogs can lead to the emergence of HBV resistant mutants of the polymerase gene. The development of drugs with a different mode of action is warranted to prevent antiviral drug resistance. Only a few non-nucleosidic molecules belonging to the family of phenylpropenamides (AT-61 & AT-130) and heteroaryldihydropyrimidines (BAY41-4109) can prevent RNA encapsidation or destabilize nucleocapsids, respectively. The sensitivity of the main nucleos(t)ide analog- resistant mutants to these inhibitors was evaluated in vitro. HepG2 stable cell lines permanently expressing wild type (WT) HBV or the main HBV mutants resistant to lamivudine and/or adefovir (rtL180M+rtM204V, rtV173L+rtL180M+rtM204V, rtM204I, rtL180M+rtM204I, rtN236T, rtA181V, rtA181V+rtN236T, rtA181T, rtA181T+rtN236T) were treated with AT-61, AT-130 or BAY-41 4109. Analysis of intracellular encapsidated viral DNA showed that all mutants were almost as sensitive to these molecules as WT HBV; indeed, the fold-resistance ranged between 0.7 and 2.3. Furthermore, the effect of a combination of either AT-61 or AT-130 with BAY41-4109, and the combination of these compounds with tenofovir was studied on wild type HBV as well as on a lamivudine and an adefovir-resistant mutant (rtL180M+M204V and rtN236T, respectively). These combinations of compounds resulted in inhibition of viral replication but showed slight antagonistic effects on the three HBV species. Based on this in vitro study, BAY-41 4109, AT-61 and AT-130 molecules that interfere with capsid morphogenesis are active against the main lamivudine- and adefovir-resistant mutants. These results suggest that targeting nucleocapsid functions may represent an interesting approach to the development of novel HBV inhibitors to prevent and combat drug resistance.
长期使用核苷（酸）类似物治疗慢性乙型肝炎可导致聚合酶基因的乙型肝炎病毒耐药突变体的出现。需要开发具有不同作用模式的药物来预防抗病毒药物耐药性。只有少数属于苯丙烯酰胺（AT-61 和 AT-130）和杂芳基二氢嘧啶（BAY41-4109）家族的非核苷分子能够分别阻止 RNA 包裹或使核衣壳不稳定。在体外评估了主要核苷（酸）类似物耐药突变体对这些抑制剂的敏感性。用 AT-61、AT-130 或 BAY-41-4109 处理永生化 HepG2 细胞株，该细胞株稳定表达野生型（WT）HBV 或对拉米夫定和/或阿德福韦耐药的主要 HBV 突变体（rtL180M+rtM204V、rtV173L+rtL180M+rtM204V、rtM204I、rtL180M+rtM204I、rtN236T、rtA181V、rtA181V+rtN236T、rtA181T、rtA181T+rtN236T）。分析细胞内包裹的病毒 DNA 表明，所有突变体对这些分子的敏感性与 WT HBV 几乎相同；实际上，耐药倍数在 0.7 到 2.3 之间。此外，还研究了 AT-61 或 AT-130 与 BAY41-4109 的联合应用，以及这些化合物与替诺福韦的联合应用对野生型 HBV 以及拉米夫定和阿德福韦耐药突变体（rtL180M+M204V 和 rtN236T）的影响。这些化合物的组合导致病毒复制的抑制，但对三种 HBV 种属显示出轻微的拮抗作用。基于这项体外研究，干扰衣壳形态发生的 BAY-41-4109、AT-61 和 AT-130 分子对主要的拉米夫定和阿德福韦耐药突变体有效。这些结果表明，针对核衣壳功能可能是开发新型 HBV 抑制剂以预防和对抗耐药性的一种有前途的方法。
