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

Abstract

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.

Zinc Metallochaperones Reactivate Mutant p53 Using an ON/OFF Switch Mechanism: A New Paradigm in Cancer Therapeutics

“…ZMC1 was administered to male BALB/c mice and plasma was harvested at various time followed by LC/MS/MS analysis for Pharmacokinetics. Quantitation of ZMC1 in plasma samples was analyzed by a validated LC/MS/MS assay as in previous report (14). The details are in Supplementary Information.…”

Influence of verapamil on pharmacokinetics of pristimerin in rats

“…Owing to the heterogeneous nature of plasma, a sample pre‐treatment procedure is often needed to remove protein and potential interfering substances before the LC‐MS/MS analysis. Currently, the most widely used biological sample preparation methods are protein precipitation, solid‐phase extraction and liquid–liquid extraction (Lin et al ., ; Soltani and Jouyban, ). All of these sample pretreatment methods were investigated to achieve good resolution and high recovery of the analyte from spiked biological matrices.…”

Copper-Binding Small Molecule Induces Oxidative Stress and Cell-Cycle Arrest in Glioblastoma-Patient-Derived Cells

“…NSC319726 was originally discovered as a selective lethal compound for cancer cell lines carrying allele specific TP53 mutation, R175H (Yu et al, 2014; Lin et al, 2015). The gain-of-function R175H mutation is one of the most frequent mutations observed in TP53 gene, contributing to various properties of transformed cancers, such as invasion, proliferation, drug resistance (Soussi and Wiman, 2007).…”