Tang Peng-Fei, Bao Su-Su, Xiao Zhong-Xiang, Xie Wei-Fei, Wu Xue-Meng, Ge Hong-Lei, Shao Chuan-Feng
Affiliated Yueqing Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, 325600, China.
Market Supervision Administration of Yueqing city, Wenzhou, Zhejiang, 325600, China.
BMC Chem. 2023 Aug 30;17(1):107. doi: 10.1186/s13065-023-01017-x.
This study establishes a UHPLC‒MS/MS method for the detection of zanubrutinib and explores its interaction with fluconazole and isavuconazole in rats.
A protein precipitation method using acetonitrile was used to prepare plasma samples using ibrutinib as an internal standard. Chromatographic separation and mass spectrometric detection of the analytes and internal standards were performed on a Shimadzu 8040 UHPLC‒MS/MS equipped with a Shim-pack velox C18 column (2.1 × 50 mm, 2.7 µm). Methanol and 0.1% formic acid-water were used as mobile phases. Intraday and interday precision and accuracy, extraction recoveries, and matrix effects of this method were determined. The linearity and sample stability of the method were assessed. Eighteen male Sprague‒Dawley (SD) rats were randomly divided into three groups with zanubrutinib (30 mg/kg) alone, zanubrutinib in combination with fluconazole (20 mg/kg) or zanubrutinib in combination with isavuconazole (20 mg/kg). Blood samples (200 µL) were collected at designated time points (ten evenly distributed time points within 12 h). The concentration of zanubrutinib was determined using the UHPLC‒MS/MS method developed in this study.
The typical fragment ions were m/z 472.15 → 290.00 for zanubrutinib and m/z 441.20 → 138.10 for ibrutinib (IS). The range of the standard curve was 1-1000 ng/mL with a regressive coefficient (R) of 0.999. The recoveries and matrix effects were 91.9-98.2% and 97.5-106.3%, respectively, at different concentration levels. The values for intra- and interday RSD% were lower than 9.8% and 5.8%, respectively. The RSD% value was less than 10.3%, and the RE% value was less than ± 4.0% under different storage conditions. Analysis of pharmacokinetic results suggested that coadministration with isavuconazole or fluconazole significantly increased the area under the curve (1081.67 ± 43.81 vs. 1267.55 ± 79.35 vs. 1721.61 ± 219.36), peak plasma concentration (332.00 ± 52.79 vs. 396.05 ± 37.19 vs. 494.51 ± 130.68), and time to peak (1.83 ± 0.41 vs. 2.00 ± 0.00 vs. 2.17 ± 0.41) compared to zanubrutinib alone.
This study provides information to understand the metabolism of zanubrutinib with concurrent use with isavuconazole or fluconazole, and further clinical trials are needed to validate the results in animals.
本研究建立了一种用于检测泽布替尼的超高效液相色谱-串联质谱（UHPLC-MS/MS）方法，并探究其在大鼠体内与氟康唑和艾沙康唑的相互作用。
采用乙腈蛋白沉淀法制备血浆样本，以依鲁替尼作为内标。在配备Shim-pack velox C18柱（2.1×50 mm，2.7 µm）的岛津8040 UHPLC-MS/MS上对分析物和内标进行色谱分离和质谱检测。以甲醇和0.1%甲酸-水作为流动相。测定该方法的日内和日间精密度与准确度、提取回收率及基质效应。评估该方法的线性和样品稳定性。将18只雄性Sprague-Dawley（SD）大鼠随机分为三组，分别单独给予泽布替尼（30 mg/kg）、泽布替尼与氟康唑（20 mg/kg）联用或泽布替尼与艾沙康唑（20 mg/kg）联用。在指定时间点（12小时内十个均匀分布的时间点）采集血样（200 µL）。采用本研究建立的UHPLC-MS/MS方法测定泽布替尼的浓度。
泽布替尼的典型碎片离子为m/z 472.15 → 290.00，依鲁替尼（内标）的典型碎片离子为m/z 441.20 → 138.10。标准曲线范围为1 - 1000 ng/mL，回归系数（R）为0.999。在不同浓度水平下，回收率和基质效应分别为91.9 - 98.2%和97.5 - 106.3%。日内和日间相对标准偏差（RSD%）值分别低于9.8%和5.8%。在不同储存条件下，RSD%值小于10.3%，相对误差（RE%）值小于±4.0%。药代动力学结果分析表明，与单独使用泽布替尼相比，与艾沙康唑或氟康唑联用显著增加了曲线下面积（1081.67±43.81 vs. 1267.55±79.35 vs. 1721.61±219.36）、血浆峰浓度（332.00±52.79 vs. 396.05±37.19 vs. 494.51±130.68）以及达峰时间（1.83±0.41 vs. 2.00±0.00 vs. 2.17±0.41）。
本研究为了解泽布替尼与艾沙康唑或氟康唑同时使用时的代谢情况提供了信息，需要进一步的临床试验来验证动物实验结果。
