Bioanalytical assay for the quantification of the tyrosine kinase inhibitor EAI045 and its major metabolite PIA in mouse plasma and tissue homogenates using liquid chromatography–tandem mass spectrometry
Susam, M. Merve; Wang, Jing; Schinkel, Alfred H.; Beijnen, Jos H.; Sparidans, Rolf W.
(2022) Biomedical Chromatography, volume 36, issue 11
(Article)
Abstract
EAI045 is a tyrosine kinase inhibitor (TKI) that targets the mutant epidermal growth factor receptor (EGFR). It was developed to control resistance to available EGFR TKIs. In this study, a major metabolite of EAI045, (5-fluoro-2-hydroxyphenyl)(1-oxo-1,3-dihydro-2H-isoindol-2-yl)acetic acid (PIA), was discovered as a hydrolysis product of the parent drug. A validated assay
... read more
for both analytes in mouse plasma and tissue homogenates from brain, kidney, liver, lung, spleen, and small intestine with content was set up using LC–MS/MS. Samples were prepared by protein precipitation with acetonitrile and with PLX4720 as internal standard. Separation was performed on a bridged ethylene hybrid C18 column by gradient elution with 0.1% v/v formic acid and methanol. Using positive electrospray, detection was performed in selected reaction monitoring mode. A linear calibration range of 2–2,000 ng/ml was used and validated for both analytes. Precision values ranged between 2.0 and 7.5% for EAI045 and between 2.2 and 12.1% for the metabolite, and accuracy values were between 91.1 and 107.6% for EAI045 and between 87.6 and 100.6% for the metabolite. Both analytes were sufficiently stable under the relevant analytical conditions. Finally, the assay was applied to analyze mouse plasma and tissue levels in a pharmacokinetic study in FVB/NRj wild-type female mice treated with oral EAI045.
show less
Download/Full Text
Keywords: EAI045, EGFR-TKI, LC–MS/MS, metabolite, mouse matrices, Analytical Chemistry, Biochemistry, Molecular Biology, Pharmacology, Drug Discovery, Clinical Biochemistry
ISSN: 0269-3879
Publisher: John Wiley and Sons Ltd
Note: Funding Information: This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. M. Merve Susam: Methodology, Validation, Formal analysis, Investigation, Writing—original draft, Visualization, Project administration. Jing Wang: Formal analysis, Investigation, Writing—review & editing, Visualization. Alfred H. Schinkel: Conceptualization, Resources, Writing—review & editing, Supervision. Jos H. Beijnen: Conceptualization, Methodology, Resources, Writing—review & editing, Supervision. Rolf W. Sparidans: Conceptualization, Methodology, Investigation, Resources, Writing—review & editing, Supervision, Project administration. Publisher Copyright: © 2022 The Authors. Biomedical Chromatography published by John Wiley & Sons Ltd.
(Peer reviewed)