Comprehensive Metabolic Profiling of Resigratinib, a Novel FGFR Inhibitor, Using Integrated LC–MS/MS and LC-Orbitrap-HRMS

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry Pub Date : 2025-09-17 DOI:10.1002/rcm.10141

Xiaoxia An, Yanting Mao, Ali Fan, Ting Ma

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引用次数: 0

Abstract

Rationale

Resigratinib, a potent fibroblast growth factor receptor (FGFR) inhibitor, is under clinical development for solid tumors such as cholangiocarcinoma. However, data on its hepatic metabolism remain limited. To support further development, this study aimed to characterize its in vitro metabolism using rat, dog, monkey, and human liver microsomes.

Methods

A sensitive and robust liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was validated for quantifying resigratinib in liver microsomes. Metabolite characterization was performed using LC coupled with benchtop Orbitrap high-resolution mass spectrometry (LC-Orbitrap-HRMS) in full-scan MS/dd-MS² and parallel reaction monitoring (PRM). This approach enabled accurate mass measurement, chemical formula assignment, and structural elucidation via MS² fragmentation interpretation.

Results

The established method exhibited excellent linearity over the concentration range of 1.0–1000 nM. Resigratinib displayed low clearance in dog (t_1/2 = 91.2 min), intermediate clearance in rat (t_1/2 = 20.2 min), and high clearance in monkey (t_1/2 = 6.8 min) and human (t_1/2 = 14.0 min) systems. Ten metabolites were identified, with M3 (bis-demethylation), M5 (O-demethylation), and M9 (N-demethylation) identified as the major metabolites. Recombinant human cytochrome P450 enzyme analysis and chemical inhibition studies indicated that CYP3A4 is the predominant enzyme responsible for resigratinib metabolism.

Conclusion

This study presents the first integrated analytical approach, combining LC–MS/MS and LC-Orbitrap-HRMS, for the in vitro metabolic assessment of resigratinib. The observed metabolic profiles provide an essential foundation for further toxicological and clinical investigations.

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基于LC-MS/MS和LC-Orbitrap-HRMS的新型FGFR抑制剂瑞格替尼综合代谢谱分析

理由：瑞格替尼是一种有效的成纤维细胞生长因子受体（FGFR）抑制剂，目前正处于用于胆管癌等实体肿瘤的临床开发阶段。然而，关于其肝脏代谢的数据仍然有限。为了支持进一步的开发，本研究旨在用大鼠、狗、猴和人的肝微粒体来表征其体外代谢。方法：建立高效液相色谱-串联质谱（LC-MS/MS）定量分析肝微粒体中瑞格瑞替尼的方法。在全扫描MS/dd-MS2和平行反应监测（PRM）中，使用LC耦合台式Orbitrap高分辨率质谱（LC-Orbitrap- hrms）进行代谢物表征。这种方法可以实现精确的质量测量，化学式分配，并通过MS2碎片解释进行结构解释。结果：所建立的方法在1.0 ~ 1000 nM的浓度范围内线性良好。瑞格替尼在犬体内清除率低（t1/2 = 91.2 min），在大鼠体内清除率中等（t1/2 = 20.2 min），在猴体内清除率高（t1/2 = 6.8 min），在人体内清除率高（t1/2 = 14.0 min）。鉴定出10种代谢物，其中M3（双去甲基化）、M5 （o -去甲基化）和M9 （n -去甲基化）是主要代谢物。重组人细胞色素P450酶分析和化学抑制研究表明，CYP3A4是瑞格替尼代谢的主要酶。结论：本研究首次采用LC-MS/MS和LC-Orbitrap-HRMS相结合的综合分析方法进行瑞格替尼体外代谢评价。观察到的代谢谱为进一步的毒理学和临床研究提供了必要的基础。

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

Rapid Communications in Mass Spectrometry 化学-分析化学

CiteScore

4.10

自引率

5.00%

发文量

219

审稿时长

2.6 months

期刊介绍： Rapid Communications in Mass Spectrometry is a journal whose aim is the rapid publication of original research results and ideas on all aspects of the science of gas-phase ions; it covers all the associated scientific disciplines. There is no formal limit on paper length ("rapid" is not synonymous with "brief"), but papers should be of a length that is commensurate with the importance and complexity of the results being reported. Contributions may be theoretical or practical in nature; they may deal with methods, techniques and applications, or with the interpretation of results; they may cover any area in science that depends directly on measurements made upon gaseous ions or that is associated with such measurements.