Lin Hongxia, Yu Xin, Eng Oliver S, Buckley Brian, Kong Ah-Ng Tony, Bertino Joseph R, Carpizo Darren R, Gounder Murugesan K
Cancer Institute of New Jersey, Rutgers, The State University of New Jersey, 195 Little Albany Street, New Brunswick, NJ, 08901, USA.
Chemical Analytical Core Laboratory, EOSHI, Rutgers, The State University of New Jersey, Piscataway, NJ, USA.
Biomed Chromatogr. 2015 Nov;29(11):1708-14. doi: 10.1002/bmc.3483. Epub 2015 May 5.
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.
ZMC1{氮杂环丁烷碳硫代酸，[1-(2-吡啶基)亚乙基]酰肼}是一种正在开发的先导化合物，作为首批靶向抗癌药物之一用于突变型p53。建立精确的定量方法是这一开发过程中不可或缺的一部分。本研究的目的是开发一种灵敏的液相色谱/串联质谱法，适用于评估ZMC1的纯度、稳定性和临床前药代动力学研究。血浆样品制备采用乙腈蛋白沉淀萃取法，ZMC1的回收率令人满意（84.2-92.8%）。使用Xterra C18柱（50×4.6 mm，3.5 µm）进行色谱分离，流动相为0.1%甲酸水溶液和乙腈，采用梯度洗脱。采用选择离子监测模式，在m/z 235.2/178.2和m/z 231.0/150.0处分别在保留时间5.2和6.3分钟时鉴定出ZMC1和内标2-氨基-6-溴苯并噻唑。该方法在人血浆中的线性范围为3.9-500.0 ng/mL，日内和日间精密度分别<5.9%和10.5%，准确度在标称值的±5.4%以内，具有可接受的重现性。该分析方法连同血浆中的基本稳定性数据和血浆结合实验为ZMC1的药代动力学研究提供了可靠的方案。这将极大地促进这种新型抗癌药物的临床前开发。
