Division of Pharmaceutical Care Sciences, Keio University Graduate School of Pharmaceutical Sciences, 1-5-30 Shibakoen, Minato-ku, Tokyo 105-8512, Japan.
Division of Pharmaceutical Care Sciences, Keio University Graduate School of Pharmaceutical Sciences, 1-5-30 Shibakoen, Minato-ku, Tokyo 105-8512, Japan; Division of Pharmaceutical Care Sciences, Center for Social Pharmacy and Pharmaceutical Care Sciences, Keio University Faculty of Pharmacy, 1-5-30 Shibakoen, Minato-ku, Tokyo 105-8512, Japan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2022 May 30;1199:123245. doi: 10.1016/j.jchromb.2022.123245. Epub 2022 Apr 4.
Reports on the therapeutic drug monitoring (TDM) of second- and third-generation epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) in non-small cell lung cancer patients are limited and are required to improve the safety of EGFR-TKI therapy. Some EGFR-TKIs have active metabolites with similar or higher potency compared with the parent compounds; thus, monitoring the parent compound as well as its active metabolites is essential for truly effective TDM. In this study, we developed and validated a method that simultaneously quantifies second- and third-generation EGFR-TKIs (afatinib, dacomitinib, and osimertinib) and the active metabolites of osimertinib, AZ5104 and AZ7550, in the human serum using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The clinical application of the method was also evaluated. The analytes were extracted from a 100 μL serum sample using a simple protein precipitation method and analyzed using LC-MS/MS. Excellent linearity of calibration curves was observed at ranges of 2.5-125.0 ng/mL for afatinib, 2.5-125.0 ng/mL for dacomitinib, 4.0-800.0 ng/mL for osimertinib, 1.0-125.0 ng/mL for AZ5104, and 2.5-125.0 ng/mL for AZ7550. The precision and accuracy were below 14.9% and within ± 14.9% of the nominal concentrations, respectively. The mean recovery was higher than 94.7% and the coefficient of variation (CV) was lower than 8.3%. The mean internal-standard normalized matrix factors ranged from 94.6 to 111.9%, and the CVs were lower than 9.7%. This analytical method met the acceptance criteria of the U.S. Food and Drug Administration guidelines. The method was also successfully applied to the analysis of 45 clinical samples; it supports the efficient and valuable analysis for TDM investigations of EGFR-TKIs.
有关第二代和第三代表皮生长因子受体酪氨酸激酶抑制剂（EGFR-TKIs）在非小细胞肺癌患者中的治疗药物监测（TDM）的报告有限，需要提高 EGFR-TKI 治疗的安全性。一些 EGFR-TKIs 具有与母体化合物具有相似或更高效力的活性代谢物；因此，监测母体化合物及其活性代谢物对于真正有效的 TDM 至关重要。在这项研究中，我们开发并验证了一种使用液相色谱-串联质谱（LC-MS/MS）同时定量人血清中第二代和第三代 EGFR-TKIs（阿法替尼、达可替尼和奥希替尼）及其奥希替尼的活性代谢物 AZ5104 和 AZ7550 的方法。还评估了该方法的临床应用。使用简单的蛋白质沉淀法从 100μL 血清样品中提取分析物，并使用 LC-MS/MS 进行分析。阿法替尼的校准曲线在 2.5-125.0ng/mL 的范围内表现出优异的线性，达可替尼的校准曲线在 2.5-125.0ng/mL 的范围内表现出优异的线性，奥希替尼的校准曲线在 4.0-800.0ng/mL 的范围内表现出优异的线性，AZ5104 的校准曲线在 1.0-125.0ng/mL 的范围内表现出优异的线性，AZ7550 的校准曲线在 2.5-125.0ng/mL 的范围内表现出优异的线性。精密度和准确度均低于 14.9%，且在名义浓度的±14.9%范围内。平均回收率高于 94.7%，变异系数（CV）低于 8.3%。平均内标归一化基质因子范围为 94.6-111.9%，CV 低于 9.7%。该分析方法符合美国食品和药物管理局指南的验收标准。该方法还成功应用于 45 个临床样本的分析；它支持对 EGFR-TKIs 的 TDM 研究进行高效和有价值的分析。
