Validation of an LC–MS/MS method for simultaneous detection of four HDAC inhibitors – belinostat, panobinostat, rocilinostat and vorinostat in mouse plasma and its application to a mouse pharmacokinetic study

Abstract

A sensitive and rapid LC-MS/MS method was developed and validated for the simultaneous quantitation of four HDAC inhibitors, namely belinostat (BST), panobinostat (PST), rocilinostat (RST) and vorinostat (VST), in mouse plasma as per regulatory guidelines. The analytes and internal standard were extracted from 50 μL mouse plasma by protein precipitation, followed by chromatographic separation using an Atlantis C column with an isocratic mobile phase comprising 0.1% formic acid-acetonitrile (25:75, v/v) at a flow rate of 0.5 mL/min within 2.5 min. Detection and quantitation were done by multiple reaction monitoring on a triple quadrupole mass spectrometer following the transitions: m/z 319 → 93, 350 → 158, 434 → 274 and 265 → 232 for BST, PST, RST and VST, respectively, in the positive ionization mode. The calibration curves were linear from 2.92 to 2921 ng/mL for BST and PST and from 1.01 to 1008 ng/mL for RST and VST with r ≥ 0.99 for all of the analytes. The intra- and inter-batch accuracy and precision (CV) across quality controls varied from 85.5 to 112% and from 2.30 to 12.5, respectively, for all of the analytes. Analytes were found to be stable under different stability conditions. The method was applied to an i.v. pharmacokinetic study in mice.

First-in-Class Hydrazide-Based HDAC6 Selective Inhibitor with Potent Oral Anti-Inflammatory Activity by Attenuating NLRP3 Inflammasome Activation

“…Blood samples were taken and analyzed for 35m using an LC-MS/MS system. As shown in Table , the in vivo t 1/2 of 35m by iv and po administration were 7.95 and 6.83 h, respectively, which are significantly longer than that of Tubastatin A (0.35 h iv and 0.86 h po) and ACY-1215 (2.62 h iv) . The AUC 0‑inf value of orally administrated 35m at 20 mg/kg is 10292 ng·h/mL, which is 76.8 times higher than that of 30 mg/kg Tubastatin A per-os and 30.1 times higher than ACY-1215 5mg/kg via iv administration .…”

“…46 The AUC 0-inf value of orally administrated 35m at 20 mg/kg is 10292 ng•h/mL, which is 76.8 times higher than that of 30 mg/kg Tubastatin A per-os 45 and 30.1 times higher than ACY-1215 5mg/kg via iv administration. 46 To our surprise, the oral bioavailability (F%) of 35m is up to 93.4%, which is 15.8 times that of Tubastatin A per-os (5.9%). 45 Therefore, 35m displays a significant improvement in the pharmacokinetic profile than the hydroxamic acid-based Tubastatin A and ACY-1215, which provides the possibility of 35m being a better orally active HDAC6 selective inhibitor.…”

A Review on Liquid Chromatography-Tandem Mass Spectrometry Methods for Rapid Quantification of Oncology Drugs

“…All of them have been classified into two categories based on the number of analytes. Forty-nine publications involved analyzing one drug (n = 1) and the remaining 23 involved analyzing multiple drugs/metabolites (n ≥ 2) are summarized in Table 1 [4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52] and Table 2 [53,54,55,56,57,58,59,60,61,62,<...>…”