A sensitive liquid chromatography-mass spectrometry bioanalytical assay for a novel anticancer candidate - ZMC1

• Published in: Biomedical chromatography Vol. 29; no. 11; pp. 1708 - 1714
• Main Authors: Lin, Hongxia, Yu, Xin, Eng, Oliver S., Buckley, Brian, Kong, Ah-Ng Tony, Bertino, Joseph R., Carpizo, Darren R., Gounder, Murugesan K.
• Format: Journal Article
• Language: English
• Published: England Blackwell Publishing Ltd 01.11.2015
• Subjects: Antineoplastic Agents - blood; Chromatography, Liquid - methods; Humans; LC-MS/MS; Limit of Detection; plasma stability; protein binding; Pyridines - blood; Reference Standards; Reproducibility of Results; Tandem Mass Spectrometry - methods; thiosemicarbazone; thiosemicarbazone; LC-MS/MS; plasma stability; protein binding
• ISSN: 0269-3879; 1099-0801
• DOI: 10.1002/bmc.3483

ZMC1 {azetidinecarbothioic acid, [1‐(2‐pyridinyl) ethylidene] hydrazide} is a lead compound being developed as one of the first mutant p53 targeted anti‐cancer drugs. Establishing a precise quantitative method is an integral component of this development. The aim of this study was to develop a sensitive LC/MS/MS assay suitable for assessing purity, stability and preclinical pharmacokinetic studies of ZMC1. Acetonitrile protein precipitation extraction was chosen for plasma sample preparation with satisfactory recovery (84.2–92.8%) for ZMC1. Chromatographic separation was achieved on an Xterra C18 column (50 × 4.6 mm, 3.5 µm) using a gradient elution with mobile phase of 0.1% formic acid in water and acetonitrile. ZMC1 and internal standard 2‐amino‐6‐bromobenzothiazole were identified using selected‐ion monitoring mode at m/z 235.2/178.2 and m/z 231.0/150.0 at retention times of 5.2 and 6.3 min, respectively. The method was validated with a linearity range of 3.9–500.0 ng/mL in human plasma and showed acceptable reproducibility with intra‐ and interday precisions <5.9 and 10.5%, and accuracy within ±5.4% of nominal values. This analytical method together with basic stability data in plasma and plasma binding experiments provides a reliable protocol for the study of ZMC1 pharmacokinetics. This will greatly facilitate the pre‐clinical development of this novel anti‐cancer drug. Copyright © 2015 John Wiley & Sons, Ltd.