Research Division of Clinical Pharmacology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
School of Pharmacy, Nanjing Medical University, Nanjing 211166, China.
Molecules. 2023 Jan 26;28(3):1205. doi: 10.3390/molecules28031205.
Ibrutinib, orelabrutinib, and zanubrutinib are all Bruton's tyrosine kinase inhibitors, which have greatly improved the treatment of B-cell malignancies. In this study, an LC-MS/MS method was developed and validated for the determination of orelabrutinib, zanubrutinib, ibrutinib, and its active metabolite dihydrodiol ibrutinib in human plasma. The Ibrutinib-d5 was used as the internal standard. Pretreatment was performed using a simple protein precipitation step using acetonitrile. The ACQUITY UPLC HSS T3 column (2.1×50 mm, 1.8 μm) was used to separate the analytes, and the run time was 6.5 min. The mobile phase consisted of acetonitrile and 10 mM of ammonium formate, which contained 0.1% formic acid. The multiple reactions' monitoring transitions were selected at m/z 428.1→411.2, 472.2→455.2, 441.1→304.2, 475.2→304.2 and 446.2→309.2 respectively for orelabrutinib, zanubrutinib, ibrutinib, dihydrodiol ibrutinib and ibrutinib-d5 using positive ion electrospray ionization. The standard curves were linear, from 0.400 to 200 ng/mL for ibrutinib and dihydrodiol ibrutinib, 1.00-500 ng/mL for orelabrutinib, and 2.00-1000 ng/mL for zanubrutinib. Selectivity, the lower limit of quantitation, precision, accuracy, matrix effect, recovery, stability, and dilution integrity all met the acceptance criteria of FDA guidance. This method was used to quantify the plasma levels of orelabrutinib, zanubrutinib, ibrutinib, and dihydrodiol ibrutinib in clinical patients.
依鲁替尼、奥瑞巴替尼和泽布替尼均为布鲁顿酪氨酸激酶抑制剂，极大地改善了 B 细胞恶性肿瘤的治疗效果。本研究建立并验证了一种 LC-MS/MS 方法，用于测定人血浆中的奥瑞巴替尼、泽布替尼、依鲁替尼及其活性代谢物二氢二醇依鲁替尼。以依鲁替尼-d5 为内标。采用乙腈简单沉淀蛋白预处理。采用 ACQUITY UPLC HSS T3 柱（2.1×50mm，1.8μm）进行分离，运行时间为 6.5min。流动相由乙腈和 10mM 甲酸铵组成，含有 0.1%甲酸。选择正离子电喷雾电离多反应监测的转换，用于奥瑞巴替尼、泽布替尼、依鲁替尼、二氢二醇依鲁替尼和依鲁替尼-d5 的 m/z 428.1→411.2、472.2→455.2、441.1→304.2、475.2→304.2 和 446.2→309.2。依鲁替尼和二氢二醇依鲁替尼的标准曲线线性范围为 0.400-200ng/mL，奥瑞巴替尼为 1.00-500ng/mL，泽布替尼为 2.00-1000ng/mL。选择性、定量下限、精密度、准确度、基质效应、提取回收率、稳定性和稀释稳定性均符合 FDA 指南的验收标准。该方法用于定量分析临床患者的奥瑞巴替尼、泽布替尼、依鲁替尼和二氢二醇依鲁替尼的血浆水平。
