Zhang Feng, Li Hong-Xin, Zhang Tian-Tian, Xiong Yuan, Wang Hao-Nan, Lu Zhen-Hui, Xiong Lei, He Yu-Qi, Ge Guang-Bo
Shanghai Frontiers Science Center for Chinese Medicine Chemical Biology, Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Shanghai Frontiers Science Center for Chinese Medicine Chemical Biology, Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai, China; School of Pharmacy, Zunyi Medical University, Zunyi, Guizhou, China.
Chem Biol Interact. 2022 Jan 5;351:109744. doi: 10.1016/j.cbi.2021.109744. Epub 2021 Nov 11.
Remdesivir, an intravenous nucleotide prodrug, has been approved for treating COVID-19 in hospitalized adults and pediatric patients. Upon administration, remdesivir can be readily hydrolyzed to form its active form GS-441524, while the cleavage of the carboxylic ester into GS-704277 is the first step for remdesivir activation. This study aims to assign the key enzymes responsible for remdesivir hydrolysis in humans, as well as to investigate the kinetics of remdesivir hydrolysis in various enzyme sources. The results showed that remdesivir could be hydrolyzed to form GS-704277 in human plasma and the microsomes from human liver (HLMs), lung (HLuMs) and kidney (HKMs), while the hydrolytic rate of remdesivir in HLMs was the fastest. Chemical inhibition and reaction phenotyping assays suggested that human carboxylesterase 1 (hCES1A) played a predominant role in remdesivir hydrolysis, while cathepsin A (CTSA), acetylcholinesterase (AchE) and butyrylcholinesterase (BchE) contributed to a lesser extent. Enzymatic kinetic analyses demonstrated that remdesivir hydrolysis in hCES1A (SHUTCM) and HLMs showed similar kinetic plots and much closed K values to each other. Meanwhile, GS-704277 formation rates were strongly correlated with the CES1A activities in HLM samples from different individual donors. Further investigation revealed that simvastatin (a therapeutic agent for adjuvant treating COVID-19) strongly inhibited remdesivir hydrolysis in both recombinant hCES1A and HLMs. Collectively, our findings reveal that hCES1A plays a predominant role in remdesivir hydrolysis in humans, which are very helpful for predicting inter-individual variability in response to remdesivir and for guiding the rational use of this anti-COVID-19 agent in clinical settings.
瑞德西韦是一种静脉注射用核苷酸前药，已被批准用于治疗住院的成人和儿童COVID-19患者。给药后，瑞德西韦可迅速水解形成其活性形式GS-441524，而羧酸酯裂解为GS-704277是瑞德西韦活化的第一步。本研究旨在确定人类中负责瑞德西韦水解的关键酶，并研究瑞德西韦在各种酶源中的水解动力学。结果表明，瑞德西韦可在人血浆以及人肝脏微粒体（HLMs）、肺微粒体（HLuMs）和肾微粒体（HKMs）中水解形成GS-704277，而瑞德西韦在HLMs中的水解速率最快。化学抑制和反应表型分析表明，人羧酸酯酶1（hCES1A）在瑞德西韦水解中起主要作用，而组织蛋白酶A（CTSA）、乙酰胆碱酯酶（AchE）和丁酰胆碱酯酶（BchE）的作用较小。酶动力学分析表明，hCES1A（SHUTCM）和HLMs中瑞德西韦的水解表现出相似的动力学曲线，且K值彼此非常接近。同时，GS-704277的形成速率与来自不同个体供体的HLM样品中的CES1A活性密切相关。进一步研究发现，辛伐他汀（一种辅助治疗COVID-19的药物）强烈抑制重组hCES1A和HLMs中瑞德西韦的水解。总体而言，我们的研究结果表明，hCES1A在人类瑞德西韦水解中起主要作用，这对于预测个体对瑞德西韦反应的变异性以及指导该抗COVID-19药物在临床环境中的合理使用非常有帮助。
