title: High-performance liquid chromatography-tandem mass spectrometry method for the determination of perampanel, a novel α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor antagonist in human plasma.
abstract: Perampanel (Fycompa(®)) is a novel α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor antagonist registered for the adjunctive treatment of patients (≥12 years) with refractory partial onset seizures. In order to support clinical trials, as well as therapeutic drug monitoring, a sensitive bioanalytical method for the determination of perampanel concentrations in human plasma was established and validated using liquid chromatography with tandem mass spectrometry. Perampanel and an internal standard were extracted from human plasma (100 μL) by liquid extraction using methyl t-butyl ether, then evaporated and reconstituted. The chromatographic separation was conducted on a C8 column with isocratic elution at a flow rate of 0.2 mL/min. The established method showed linearity in the range 0.25-200 ng/mL with correlation coefficients of >0.99 that could be extended 10-fold as validated by dilution integrity analyses. No significant endogenous peaks were detected in the elution of analytes in blank human plasma and no significant matrix effect was observed. The intra- and inter-batch reproducibility analyses demonstrated accuracy and precision within the acceptance criteria. To check the impact of anti-epileptic drugs on the perampanel assay, accuracy, precision, and specificity were assessed in the presence of 14 anti-epileptic drugs. No anti-epileptic drugs at clinically relevant levels showed a significant impact on the perampanel assay.
authors.affiliation: Drug Metabolism and Pharmacokinetics, Biopharmaceutical Assessment Core Function Unit, Eisai Co., Ltd., 1-3, 5-chome, Tokodai, Tsukuba-shi, Ibaraki 300-2635, Japan. Electronic address: y2-mano@hhc.eisai.co.jp.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Mano
authors.affiliation: Drug Metabolism and Pharmacokinetics, Biopharmaceutical Assessment Core Function Unit, Eisai Co., Ltd., 1-3, 5-chome, Tokodai, Tsukuba-shi, Ibaraki 300-2635, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Takenaka
authors.affiliation: Drug Metabolism and Pharmacokinetics, Biopharmaceutical Assessment Core Function Unit, Eisai Co., Ltd., 1-3, 5-chome, Tokodai, Tsukuba-shi, Ibaraki 300-2635, Japan.
authors.forename: Kazutomi
authors.initials: K
authors.lastname: Kusano
mesh_terms: D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D009570:Nitriles; D010949:Plasma / Q000737:chemistry*; D011728:Pyridones / Q000097:blood* / Q000737:chemistry*; D018091:Receptors, AMPA / Q000037:antagonists & inhibitors*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid / Q000378:metabolism*
chemical_list: D009570:Nitriles; D011728:Pyridones; D018091:Receptors, AMPA; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; C551441:perampanel
keywords: Epilepsy; Perampanel; Pharmacokinetics; Plasma; Validation