Activation of Tenofovir Alafenamide and Sofosbuvir in the Human Lung and Its Implications in the Development of Nucleoside/Nucleotide Prodrugs for Treating SARS-CoV-2 Pulmonary Infection.
Activation of Tenofovir Alafenamide and Sofosbuvir in the Human Lung and Its Implications in the Development of Nucleoside/Nucleotide Prodrugs for Treating SARS-CoV-2 Pulmonary Infection.
复制标题
DOI：
10.3390/pharmaceutics13101656
复制
发表时间：
2021-10-11
期刊：
影响因子：
5.4
通讯作者：
Zhu HJ
中科院分区：
文献类型：
--
作者：
Li J;Liu S;Shi J;Zhu HJ
ProTide 技术是设计核苷/核苷酸类似物前药的强大工具。 ProTide 前药设计可改善细胞通透性并增强细胞内活化。 ProTide 酯键的水解是前药细胞内激活效率和最终抗病毒功效的决定因素。水解由催化活性和活化酶的丰度决定。抗病毒药物替诺福韦艾拉酚胺 (TAF) 和索磷布韦 (SBV) 是典型的 ProTides。 TAF 和 SBV 也被提议用于治疗 COVID-19 患者。然而，这两种前药在肺中激活的机制仍不确定。在本研究中，我们使用基于活性的蛋白质分析测定来分析人肺 S9 组分中丝氨酸水解酶的催化活性。我们使用人肺和肝脏 S9 组分和纯化酶评估了 TAF 和 SBV 的水解。结果表明，CatA和CES1参与了两种前药在人肺中的水解。更具体地说，CatA 对 TAF 的水解活性比 SBV 高出近 4 倍，而 CES1 对 TAF 的水解活性略低于 SBV。总体而言，TAF 在人肺 S9 中的水解率比 SBV 高近 4 倍。我们利用从文献中提取的蛋白质组学数据进一步分析了人肺、肝和这两种组织的原代细胞中 CatA 和 CES1 的蛋白表达水平。 CatA 与 CES1 的相对蛋白质丰度在人肺和原代人气道上皮细胞中比在人肝脏和原代人肝细胞中高得多。研究结果表明，TAF 对 CatA 介导的水解高度敏感，导致 TAF 在人肺中有效水解，这表明在设计用于治疗呼吸道病毒感染的抗病毒酯前药时，CatA 可以用作目标激活酶。
ProTide technology is a powerful tool for the design of nucleoside/nucleotide analog prodrugs. ProTide prodrug design improves cell permeability and enhances intracellular activation. The hydrolysis of the ester bond of a ProTide is a determinant of the intracellular activation efficiency and final antiviral efficacy of the prodrug. The hydrolysis is dictated by the catalytic activity and abundance of activating enzymes. The antiviral agents tenofovir alafenamide (TAF) and sofosbuvir (SBV) are typical ProTides. Both TAF and SBV have also been proposed to treat patients with COVID-19. However, the mechanisms underlying the activation of the two prodrugs in the lung remain inconclusive. In the present study, we profiled the catalytic activity of serine hydrolases in human lung S9 fractions using an activity-based protein profiling assay. We evaluated the hydrolysis of TAF and SBV using human lung and liver S9 fractions and purified enzymes. The results showed that CatA and CES1 were involved in the hydrolysis of the two prodrugs in the human lung. More specifically, CatA exhibited a nearly 4-fold higher hydrolytic activity towards TAF than SBV, whereas the CES1 activity on hydrolyzing TAF was slightly lower than that for SBV. Overall, TAF had a nearly 4-fold higher hydrolysis rate in human lung S9 than SBV. We further analyzed protein expression levels of CatA and CES1 in the human lung, liver, and primary cells of the two tissues using proteomics data extracted from the literature. The relative protein abundance of CatA to CES1 was considerably higher in the human lung and primary human airway epithelial cells than in the human liver and primary human hepatocytes. The findings demonstrated that the high susceptivity of TAF to CatA-mediated hydrolysis resulted in efficient TAF hydrolysis in the human lung, suggesting that CatA could be utilized as a target activating enzyme when designing antiviral ester prodrugs for the treatment of respiratory virus infection.
登录
查看更多内容
影响因子：
4.8
作者：
Murakami, Eisuke;Tolstykh, Tatiana;Furman, Phillip A.
通讯作者：
Furman, Phillip A.
影响因子：
4.4
作者：
Foster MW;Gwinn WM;Kelly FL;Brass DM;Valente AM;Moseley MA;Thompson JW;Morgan DL;Palmer SM
通讯作者：
Palmer SM
影响因子：
4.4
作者：
Chien M;Anderson TK;Jockusch S;Tao C;Li X;Kumar S;Russo JJ;Kirchdoerfer RN;Ju J
通讯作者：
Ju J
影响因子：
4.6
作者：
Jockusch S;Tao C;Li X;Chien M;Kumar S;Morozova I;Kalachikov S;Russo JJ;Ju J
通讯作者：
Ju J
影响因子：
4.9
作者：
Birkus, Gabriel;Wang, Ruth;McDermott, Martin
通讯作者：
McDermott, Martin