Metabolic Profiling of Canertinib: A Comprehensive Cross-Species Investigation Using Advanced UPLC-MS/MS and...

Area of Science:

• Pharmacology and Drug Metabolism
• Analytical Chemistry
• Biochemistry

Background:

• Canertinib is an EGFR tyrosine kinase inhibitor investigated for leukemia and non-small cell lung cancer.
• Understanding drug metabolism is crucial for predicting efficacy and toxicity.
• Species-specific metabolic differences can impact drug development and clinical outcomes.

Purpose of the Study:

• To develop and validate a UPLC-MS/MS method for quantifying canertinib metabolic stability in liver microsomes.
• To characterize the metabolites of canertinib across different species.
• To investigate interspecies variations in canertinib metabolism.

Main Methods:

• Ultra-Performance Liquid Chromatography-tandem Mass Spectrometry (UPLC-MS/MS) for quantitative analysis.
• Liquid Chromatography-Orbitrap High-Resolution Mass Spectrometry (LC-Orbitrap-HRMS) for metabolite identification.
• In vitro incubation with liver microsomes from human, rat, and monkey.
• Post-acquisition data mining techniques including high-resolution extracted ion chromatograms and multimass defect filtering.

Main Results:

• The UPLC-MS/MS method demonstrated excellent linearity (10-1000 nM) for high-throughput screening.
• Canertinib exhibited significant species-dependent intrinsic clearance (CL_{in vitro, mic}): human < rat < monkey.
• Sixteen NADPH-dependent metabolites and one GSH conjugate were structurally characterized, with specific metabolites identified as human- or monkey-specific.
• Metabolic pathways included oxidative defluorination, O-dealkylation, oxidative deamination, piperidine ring opening, lactam formation, and GSH conjugation.

Conclusions:

• This study presents the first cross-species metabolic investigation of canertinib.
• Significant interspecies metabolic disparities were observed, highlighting the importance of comparative metabolism studies.
• The elucidated metabolic framework provides critical insights into canertinib's pharmacological activity and potential toxicity profiles, aiding further drug development.