A Novel HPLC-MS/MS Method for the Intracellular Quantification of the Active Triphosphate Metabolite of Remdesivir: GS-443902.

IF 4.4 Q1 TOXICOLOGY

Journal of Xenobiotics Pub Date : 2025-07-03 DOI:10.3390/jox15040107

Alice Palermiti, Amedeo De Nicolò, Miriam Antonucci, Sara Soloperto, Martina Billi, Alessandra Manca, Jessica Cusato, Giorgia Menegatti, Mohammed Lamorde, Andrea Calcagno, Catriona Waitt, Antonio D'Avolio

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引用次数: 0

Abstract

Background: Remdesivir (RDV) is a broad-spectrum antiviral prodrug, which is rapidly metabolized in vivo within cells to the pharmacologically active triphosphate metabolite, GS-443902. On the other hand, the dephosphorylated metabolite GS-441524 is the main form detected in plasma. RDV acts against RNA viruses, and it was the first antiviral drug to receive EMA and FDA approval for treating COVID-19. Nevertheless, its intracellular pharmacokinetics in real life are poorly explored, particularly due to technical challenges.

Methods: The aim of this study was to validate an HPLC-MS/MS method for the direct quantification of GS-443902 in peripheral blood mononuclear cells (PBMCs) with a chromatographic separation of 15 min.

Results: The method was validated following EMA and FDA guidelines in terms of sensitivity, specificity, accuracy, precision, matrix effect, recovery, carryover, and stability, and then applied to PBMC isolates from a small cohort of patients with severe COVID-19 who received RDV.

Conclusions: This work represents the first method for the direct quantification of GS-443902 in PBMCs, with possible future application to intracellular pharmacokinetic studies in different scenarios, such as new oral prodrugs or drug-drug interaction studies.

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一种新的高效液相色谱-质谱/质谱法测定瑞德西韦活性三磷酸代谢物GS-443902

背景：雷姆德西韦（Remdesivir， RDV）是一种广谱抗病毒前药，在体内细胞内迅速代谢为具有药理活性的三磷酸代谢物GS-443902。另一方面，去磷酸化代谢物GS-441524是血浆中检测到的主要形式。RDV作用于RNA病毒，是首个获得EMA和FDA批准用于治疗COVID-19的抗病毒药物。然而，其在现实生活中的细胞内药代动力学研究很少，特别是由于技术上的挑战。结果：该方法在敏感性、特异性、准确性、精密度、基质效应、回收率、携带性和稳定性等方面均符合EMA和FDA的指导方针，并应用于一小群接受RDV治疗的重症COVID-19患者的PBMC分离株。结论：本工作是首个直接定量GS-443902在PBMCs中的方法，未来可能应用于不同情况下的细胞内药代动力学研究，如新的口服前药或药物-药物相互作用研究。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Xenobiotics TOXICOLOGY-

CiteScore

5.30

自引率

1.70%

发文量

审稿时长

10 weeks

期刊介绍： The Journal of Xenobiotics publishes original studies concerning the beneficial (pharmacology) and detrimental effects (toxicology) of xenobiotics in all organisms. A xenobiotic (“stranger to life”) is defined as a chemical that is not usually found at significant concentrations or expected to reside for long periods in organisms. In addition to man-made chemicals, natural products could also be of interest if they have potent biological properties, special medicinal properties or that a given organism is at risk of exposure in the environment. Topics dealing with abiotic- and biotic-based transformations in various media (xenobiochemistry) and environmental toxicology are also of interest. Areas of interests include the identification of key physical and chemical properties of molecules that predict biological effects and persistence in the environment; the molecular mode of action of xenobiotics; biochemical and physiological interactions leading to change in organism health; pathophysiological interactions of natural and synthetic chemicals; development of biochemical indicators including new “-omics” approaches to identify biomarkers of exposure or effects for xenobiotics.