Cell-Dependent Activation of ProTide Prodrugs and Its Implications in Antiviral Studies.
Cell-Dependent Activation of ProTide Prodrugs and Its Implications in Antiviral Studies.
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DOI：
10.1021/acsptsci.3c00050
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发表时间：
2023-07
影响因子：
6
通讯作者：
Yue-ting Liu;Shuxin Sun;Jiapeng Li;Weiwen Wang;Hao‐Jie Zhu
中科院分区：
文献类型：
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作者：
Yue-ting Liu;Shuxin Sun;Jiapeng Li;Weiwen Wang;Hao‐Jie Zhu
磷酰胺酯前药设计是改善细胞渗透性和增强核苷酸抗病毒类似物的细胞内活化的有力工具。先前的体外研究表明，磷酰胺酯前药的活化在不同的细胞系中不同。在本研究中，我们研究了两种抗病毒前体药物替诺福韦艾拉酚胺（TAF）和索非布韦（SOF）在5种抗病毒研究中常用的细胞系，即Vero E6，Huh-7，Calu-3，A549和Caco-2中的活化概况。我们发现TAF和SOF以细胞依赖性的方式被激活，其中Vero E6是激活前药的效率最低的细胞系，Huh-7是激活前药的效率最高的细胞系。我们还证明了TAF的激活率明显高于SOF。我们进一步分析了激活酶羧酸酯酶1，组织蛋白酶A，组氨酸三联体核苷酸结合蛋白1，以及相关的药物转运蛋白P-糖蛋白和有机阴离子转运多肽1B 1和1B 3的蛋白表达在细胞系中使用蛋白质组学数据提取的文献和蛋白质组数据库。结果显示，细胞系之间的酶和转运蛋白的表达模式存在显着差异，这可能部分有助于观察到的TAF和SOF的细胞依赖性激活。这些发现突出了细胞系之间的活化酶和转运蛋白的丰度的可变性，并强调了选择适当的细胞系用于评估核苷/核苷酸前药的抗病毒功效的重要性。
The ProTide prodrug design is a powerful tool to improve cell permeability and enhance the intracellular activation of nucleotide antiviral analogues. Previous in vitro studies showed that the activation of ProTide prodrugs varied in different cell lines. In the present study, we investigated the activation profiles of two antiviral prodrugs tenofovir alafenamide (TAF) and sofosbuvir (SOF) in five cell lines commonly used in antiviral research, namely, Vero E6, Huh-7, Calu-3, A549, and Caco-2. We found that TAF and SOF were activated in a cell-dependent manner with Vero E6 being the least efficient and Huh-7 being the most efficient cell line for activating the prodrugs. We also demonstrated that TAF was activated at a significantly higher rate than SOF. We further analyzed the protein expressions of the activating enzymes carboxylesterase 1, cathepsin A, histidine triad nucleotide-binding protein 1, and the relevant drug transporters P-glycoprotein and organic anion-transporting polypeptides 1B1 and 1B3 in the cell lines using the proteomics data extracted from the literature and proteome database. The results revealed significant differences in the expression patterns of the enzymes and transporters among the cell lines, which might partially contribute to the observed cell-dependent activation of TAF and SOF. These findings highlight the variability of the abundance of activating enzymes and transporters between cell lines and emphasize the importance of selecting appropriate cell lines for assessing the antiviral efficacy of nucleoside/nucleotide prodrugs.