Cell-permeable artificial zinc-finger proteins as potent antiviral drugs for human papillomaviruses
Cell-permeable artificial zinc-finger proteins as potent antiviral drugs for human papillomaviruses
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DOI：
10.1007/s00705-008-0125-7
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发表时间：
2008-07-01
影响因子：
2.7
通讯作者：
Sera, Takashi
中科院分区：
文献类型：
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作者：
Mino, Takashi;Mori, Tomoaki;Sera, Takashi
人乳头瘤病毒（HPV）是重要的药物靶点之一，因为高危类型的感染会导致浸润性宫颈癌。然而，迄今为止，尚未开发出针对HPV的有效抗病毒药物。在本研究中，我们通过将先前生成的用于抑制 HPV-18 DNA 复制的 AZP 与细胞穿透肽 (CPP) 融合，构建了细胞可渗透的人工锌指蛋白 (AZP)，作为针对 HPV 的新型抗病毒药物的候选药物。我们证实，当这些 CPP-AZP 融合体添加到培养基中时，会降低瞬时复制测定中的复制率。特别是，含有 9 聚体精氨酸作为 CPP 的 250 nM CPP-AZP（指定为 AZP-R9）可将 HPV-18 DNA 复制减少至对照的 3%，并且 50% 有效浓度（EC50）< 31 nM。此外，细胞毒性测定显示AZP-R9的50%抑制浓度（IC50）>10μM。因此，选择性指数（定义为IC50/EC50）>300，优于抗HPV病毒西多福韦。因此，我们的结果表明，细胞渗透性 AZP 可以作为有效的基于蛋白质的抗病毒药物。
Human papillomavirus (HPV) is one of the important pharmaceutical targets because infection of the high-risk types causes invasive cervical cancer. However, effective antiviral drugs for HPV have not been developed so far. In the present study, we constructed cell-permeable artificial zinc-finger proteins (AZPs) by fusing an AZP previously generated for inhibition of HPV-18 DNA replication with a cell-penetrating peptide (CPP) as candidates for new antiviral drugs against HPV. We confirmed that these CPP-AZP fusions reduced the replication rate in transient replication assays when added to the culture medium. In particular, 250 nM CPP-AZP (designated AZP-R9) containing a 9-mer of arginine as the CPP reduced HPV-18 DNA replication to 3% of that of a control, and the 50% effective concentration (EC50) was < 31 nM. Furthermore, a cytotoxicity assay revealed that the 50% inhibitory concentration (IC50) of AZP-R9 was > 10 mu M. Therefore, the selectivity index, defined as IC50/EC50, was > 300, which is better than that of the antiviral cidofovir for HPVs. Thus, our results demonstrate that cell-permeable AZPs could serve as potent protein-based antiviral drugs.