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Updated: Jan 17, 2026
An Integrated Raman Spectroscopy and Mass Spectrometry Platform to Study Single-Cell Drug Uptake, Metabolism, and Effects
Published on: January 9, 2020
Comprehensive Metabolic Profiling of Resigratinib, a Novel FGFR Inhibitor, Using Integrated LC-MS/MS and
Xiaoxia An1, Yanting Mao1, Ali Fan2
1Department of Pharmacy, Children's Hospital Affiliated to Zhengzhou University, Zhengzhou, China.
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 (t1/2 = 91.2 min), intermediate clearance in rat (t1/2 = 20.2 min), and high clearance in monkey (t1/2 = 6.8 min) and human (t1/2 = 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.
Insights
This study characterized the in vitro metabolism of resigratinib, a fibroblast growth factor receptor (FGFR) inhibitor, in liver microsomes. CYP3A4 was identified as the primary enzyme responsible for resigratinib metabolism, with ten metabolites identified.
Area of Science:
- Pharmacology
- Drug Metabolism
- Biochemistry
Background:
- Resigratinib is a potent fibroblast growth factor receptor (FGFR) inhibitor investigated for solid tumors like cholangiocarcinoma.
- Limited data exists regarding the hepatic metabolism of resigratinib, necessitating further investigation for its clinical development.
Purpose of the Study:
- To characterize the in vitro hepatic metabolism of resigratinib.
- To identify major metabolites and the primary enzymes involved in resigratinib metabolism across different species.
Main Methods:
- Validated a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for resigratinib quantification in liver microsomes.
- Employed LC coupled with Orbitrap high-resolution mass spectrometry (LC-Orbitrap-HRMS) for metabolite identification and structural elucidation.
- Utilized recombinant cytochrome P450 enzymes and chemical inhibition studies to determine the metabolic enzymes involved.
Main Results:
- Resigratinib exhibited low clearance in dogs and high clearance in monkeys and humans.
- Ten metabolites were identified, with M3 (bis-demethylation), M5 (O-demethylation), and M9 (N-demethylation) being the major ones.
- Cytochrome P450 3A4 (CYP3A4) was identified as the predominant enzyme responsible for resigratinib metabolism.
Conclusions:
- This study provides the first integrated analytical approach for the in vitro metabolic assessment of resigratinib.
- The identified metabolic profiles and pathways are crucial for future toxicological and clinical studies of resigratinib.
