Key Metabolic Enzymes Involved in Remdesivir Activation in Human Lung Cells

作者：Ruidong Li, Albert Liclican, Yili Xu, Jared Pitts, Congrong Niu, Jingyu Zhang, Cynthia Kim, Xiaofeng Zhao, Daniel Soohoo, Darius Babusis, Qin Yue, Bin Ma, Bernard P. Murray, Raju Subramanian, Xuping Xie, Jing Zou, John P. Bilello, Li Li, Brian E. Schultz, Roman Sakowicz, Bill J. Smith, Pei‐Yong Shi, Eisuke Murakami, Joy Y. Feng · 发表于：Antimicrobial Agents and Chemotherapy · 年份：2021 · DOI：10.1128/aac.00602-21 · 被引用次数：63 · 研究领域：RNA modifications and cancer、interferon and immune responses、Respiratory viral infections research

Remdesivir (RDV; GS-5734, Veklury), the first FDA-approved antiviral to treat COVID-19, is a single-diastereomer monophosphoramidate prodrug of an adenosine analogue. RDV is taken up in the target cells and metabolized in multiple steps to form the active nucleoside triphosphate (TP) (GS-443902), which, in turn, acts as a potent and selective inhibitor of multiple viral RNA polymerases. In this report, we profiled the key enzymes involved in the RDV metabolic pathway with multiple parallel approaches: (i) bioinformatic analysis of nucleoside/nucleotide metabolic enzyme mRNA expression using public human tissue and lung single-cell bulk mRNA sequence (RNA-seq) data sets, (ii) protein and mRNA quantification of enzymes in human lung tissue and primary lung cells, (iii) biochemical studies on the catalytic rate of key enzymes, (iv) effects of specific enzyme inhibitors on the GS-443902 formation, and (v) the effects of these inhibitors on RDV antiviral activity against SARS-CoV-2 in cell culture. Our data collectively demonstrated that carboxylesterase 1 (CES1) and cathepsin A (CatA) are enzymes involved in hydrolyzing RDV to its alanine intermediate MetX, which is further hydrolyzed to the monophosphate form by histidine triad nucleotide-binding protein 1 (HINT1). The monophosphate is then consecutively phosphorylated to diphosphate and triphosphate by cellular phosphotransferases. Our data support the hypothesis that the unique properties of RDV prodrug not only allow lung-spe...