An LC–MS/MS method for determination ofthe bromodomain inhibitor ZEN-3694 and itsmetabolite ZEN-3791 in human plasma: supplementary data

We have developed and validated a novel LC–MS/MS method for the simultaneous quantification of ZEN-3694 and its active metabolite ZEN-3791 in human plasma after protein precipitation. Stable isotope labeledversions were used as internal standards. Chromatographic separation was achieved on a KinetexC18 column using 0.1% formic acid in H2O and 0.1% formic acid in MeOH as mobile phases. Detectionwas performed via positive electrospray ionization mode with multiple reaction monitoring. The assayexhibited linearity in the concentration range of 5–5000 ng/ml for both analytes. Intra- and inter-assayprecision and accuracy were within ±11%. ZEN-3694 and ZEN-3791 recoveries were between 93 and 105%.This LC–MS/MS assay is an essential tool to study ZEN-3694 in an ongoing clinical trial (NCT04840589).

本研究建立并验证了一种新型液相色谱-串联质谱（LC–MS/MS）分析方法，可通过蛋白沉淀法同时定量人血浆中的ZEN-3694及其活性代谢物ZEN-3791。本方法采用稳定同位素标记物作为内标。色谱分离在Kinetex C18色谱柱上完成，流动相为含0.1%甲酸的水溶液与含0.1%甲酸的甲醇溶液。检测采用正离子电喷雾电离模式结合多反应监测技术。该分析方法对两种分析物的线性浓度范围均为5~5000 ng/ml。批内及批间精密度与准确度均控制在±11%以内。ZEN-3694与ZEN-3791的回收率介于93%至105%之间。本LC–MS/MS分析方法是一项核心工具，可用于正在进行的临床试验（NCT04840589）中对ZEN-3694的相关研究。