RESUMO
A sensitive and rapid liquid chromatography tandem mass spectrometry(LC-MS/MS)method was established for the quantification of total and unbound concentrations of LY3214996,an extracellular signal-regulated kinase inhibitor;abemaciclib,a cyclin-dependent kinase 4/6 inhibitor;and abemaciclib active metabolites,M2 and M20,in human plasma,brain tumor,and cerebrospinal fluid samples.The method was validated over a concentration range of 0.2-500 nM within a total run time of 3.8 min using isocratic elution on a Kinetex? Fs column.Detection was performed on a Sciex QTRAP 6500+mass spectrometer employing multiple reaction monitoring mode under positive electrospray ionization.The intra-and inter-batch accuracy as well as the precision of the method for all matrices was within±20％and≤20％at the lower limit of quantification,and within±15％and≤15％for other quality control levels for all analytes.The unbound fractions of drugs and metabolites in spiked and patient samples were determined using an optimized equilibrium dialysis.The validated method was successfully applied in a phase 0/2 clinical trial to assess the central nervous system penetration of LY3214996 and abemaciclib.
RESUMO
A rapid,sensitive,and robust reversed-phase liquid chromatography with tandem mass spectrometrymethod was developed and validated for the determination of total and unbound ceritinib,a secondgenerationALK inhibitor,in patient plasma and brain tumor tissue samples.Sample preparation involvedsimple protein precipitation with acetonitrile.Chromatographic separation was achieved on a Waters ACQUITYUPLC BEH C18 column using a 4-min gradient elution consisting of mobile phase A(0.1% formic acidinwater)andmobile phase B(0.1% formic acid in acetonitrile),at a flow rate of 0.4 mL/min.Ceritinib and theinternal standard([13C6]ceritinib)were monitored using multiple reaction monitoring mode under positiveelectrospray ionization.The lower limit of quantitation(LLOQ)was 1 nM of ceritinib in plasma.The calibrationcurve was linear over ceritinib concentration range of 1–2000 nM in plasma.The intra-and interdayprecision and accuracy were within the generally accepted criteria for bioanalytical method(<15%).The method was successfully applied to assess ceritinib brain tumor penetration,as assessed by the unbounddrug brain concentration to unbound drug plasma concentration ratio,in patients with brain tumors.