In vitro and in vivo pharmacokinetics and metabolism of MK‐8353 by liquid chromatography combined with diode array detector and Q‐Exactive‐Orbitrap tandem mass spectrometry

Abstract

HighlightsAn high resolution LC/MS method for the determination of MK‐8353 in rat plasma.Pharmacokinetic profiles were investigated.In vitro and in vivo metabolites were structurally characterized.Metabolic pathways referred to dealkylation, oxygenation and lactam formation. ABSTRACT In this study, a simple and sensitive quantitation method based on liquid chromatography combined with diode array detector and Q‐Exactive‐Orbitrap tandem mass spectrometry was developed for the determination of MK‐8353 in rat plasma. The chromatographic separation was carried out on a Waters ACQUITY BEH C18 column by using water containing 1 mM ammonium acetate and acetonitrile containing 0.1% formic acid as mobile phase. The developed assay was linear (r > 0.999) over the concentration range of 1–1000 ng/mL. The selectivity, precision, accuracy, recovery, matrix effects and stability were all within the required limits. The validated assay has been further applied to the pharmacokinetic study of MK‐8353 in rat after intravenous and oral administration, which revealed that MK‐8353 showed low clearance and satisfactory bioavailability. More importantly, the metabolites of MK‐8353 present in rat plasma, RLM, DLM and HLM were identified and profiled. Under the current conditions, a total of 10 metabolites were detected and their chemical structures were proposed in terms of the accurate masses and their fragment ions. Our results revealed that MK‐8353 was metabolized mainly through dealkylation, demethylation, depropylation, oxygenation, sulfur oxidation and formation of lactam. Compared with animal species, no human‐specific metabolite was found in HLM. This study provides overall in vitro and in vivo profiles of MK‐8353, which is of great help in understanding its PK/PD profiles and in predicting human pharmacokinetic profiles.