Background: EAI045 is the fourth-generation epidermal growth factor receptor tyrosine kinase inhibitor (EGFR TKI), which can overcome acquired resistance to the third-generation EGFR TKIs and is the first allosteric inhibitor that targets T790M and C797S EGFR mutants.
Methods: A rapid and sensitive LC-MS/MS method was established and validated for the quantification of EAI045 in rat plasma. Chromatographic separation was carried out at 25°C on a Hypersil GOLD C18 column (50 mm × 2.1 mm, 3 μm) and eluted on a gradient mobile phase of water (containing 0.1% formic acid) and acetonitrile at a flow rate of 0.5 mL/min. The mass spectrometer was operated in the positive ESI mode and selected reaction monitoring mode.
Results: The assay was validated over a concentration range of 1.0 - 1000 ng/mL for EAI045 with a lower limit of quantification (LLOQ) of 1.0 ng/mL. The intra- and inter-batch accuracy for the EAI045 ranged from 92.25% to 97.18% and 95.94% to 102.69%, and the intra- and inter-batch precision for the EAI045 ranged from 1.41% to 4.57% and 5.18% to 6.37%, respectively. The extraction recovery, matrix effect and stability met all requirements of the guidelines for bioanalytical method validation.
Conclusion: The rapid and sensitive LC-MS/MS method was successfully applied in a pharmacokinetic study of EAI045 following oral administration (5 mg/kg) to rats.
[http://dx.doi.org/10.1021/acs.jmedchem.5b01082] [PMID: 26275028]
[http://dx.doi.org/10.2174/1573412912666160527125937] [PMID: 28845150]