Cui Xinge, Zheng Xin, Jiang Ji, Tan Fenlai, Ding Lieming, Hu Pei
Clinical Pharmacology Research Center, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, No. 41. Damucang Hutong, Xicheng District, Beijing, 100032, China.
Betta Pharmaceuticals Co., Ltd, No. 589 Hongfeng Road, Yuhang Economic and Technological Development Area, Hangzhou, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Nov 15;1068-1069:33-40. doi: 10.1016/j.jchromb.2017.10.017. Epub 2017 Oct 8.
BPI-9016M is a novel dual-target small-molecule inhibitor targeting c-Met and AXL, which was developed by Betta Pharmaceuticals Co., Ltd (Hangzhou, China). It has great potential in the treatment of advanced cancer. A high throughput quantitation method, based on liquid chromatography-tandem mass spectrometry, was developed and validated for the simultaneous determination of BPI-9016M and its main metabolite, M1 and M2-2, in human plasma with a sample preparation method of precipitation of protein. Liquid chromatographic separation was performed with a gradient elution of formic acid-10mM ammonium acetate aqueous solution (1:1000, v/v) and acetonitrile at a flow rate of 0.4mL/min within 2.2min. A Waters ACQUITY UPLC BEH C18 column (1.7μm, 2.1×50mm) was chosen, of which the temperature was set to be 40°C. Mass spectrometric detection, which were achieved in positive mode, were performed by multiple reaction monitoring with SCIEX API 5500 Qtrap equipped with an ESI ion source. This method showed good linearity, accuracy and precision in the range of 0.4-200ng/mL for BPI-9016M and 0.8-800ng/mL for M1 and M2-2, with high recovery and slight matrix effect for all analytes. And under the conditions same as stability assessments in method validation, the three analytes stayed stable during the entire destiny of a clinical sample from the collection of whole blood to the analysis of plasma by this method. The validated method was successfully applied to a first-in-human, dose-escalation phase I clinical trial in Chinese advanced solid tumor patients for the pharmacokinetic research of BPI-9016M tablet after oral administration. The concentration-time curves of BPI-9016M, M1, M2-2 were detailly captured with good veracity. And according to the results of hemolysis assessment, plasma concentrations of analytes in hemolyzed plasma samples could be reported normally without label.
BPI-9016M是一种新型双靶点小分子抑制剂，靶向c-Met和AXL，由贝达药业股份有限公司（中国杭州）研发。它在晚期癌症治疗方面具有巨大潜力。建立了一种基于液相色谱-串联质谱的高通量定量方法，并进行了验证，用于同时测定人血浆中的BPI-9016M及其主要代谢物M1和M2-2，采用蛋白质沉淀的样品制备方法。液相色谱分离采用甲酸-10mM醋酸铵水溶液（1:1000，v/v）和乙腈梯度洗脱，流速为0.4mL/min，在2.2分钟内完成。选用沃特世ACQUITY UPLC BEH C18柱（1.7μm，2.1×50mm），柱温设定为40°C。质谱检测采用正离子模式，通过配备电喷雾离子源的SCIEX API 5500 Qtrap进行多反应监测。该方法在0.4-200ng/mL范围内对BPI-9016M以及0.8-800ng/mL范围内对M1和M2-2表现出良好的线性、准确性和精密度，所有分析物回收率高且基质效应小。在与方法验证中的稳定性评估相同的条件下，这三种分析物在临床样品从全血采集到血浆分析的整个过程中保持稳定。该验证方法成功应用于中国晚期实体瘤患者的首次人体剂量递增I期临床试验，用于口服BPI-9016M片剂后的药代动力学研究。详细准确地捕捉到了BPI-9016M、M1、M2-2的浓度-时间曲线。根据溶血评估结果，溶血血浆样品中分析物的血浆浓度无需标记即可正常报告。
