Service and Laboratory of Clinical Pharmacology, Department of Laboratory Medicine and Pathology, University Hospital and University of Lausanne, Lausanne, Switzerland.
Service and Laboratory of Clinical Pharmacology, Department of Laboratory Medicine and Pathology, University Hospital and University of Lausanne, Lausanne, Switzerland.
J Chromatogr B Analyt Technol Biomed Life Sci. 2023 Nov 15;1230:123917. doi: 10.1016/j.jchromb.2023.123917. Epub 2023 Oct 29.
Janus kinase inhibitors (JAKi) are oral small molecules used in the treatment of a broad spectrum of autoimmune and myeloproliferative diseases. JAKi exhibit significant intra- and inter-individual pharmacokinetic variabilities, due to fluctuations in compliance with oral treatments and their metabolism essentially driven by cytochrome P450 enzymes. Intrinsically, JAKi have dose-response relationship and narrow therapeutic index: therapeutic drug monitoring (TDM) is expected to optimize and adapt their dosage regimen in order to resolve problems of efficacy and tolerance linked to dose and safety. A sensitive analytical method using multiplex high-performance liquid-chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) was developed and validated for the simultaneous quantification in plasma of the 6 major currently used JAKi, namely abrocitinib, baricitinib, fedratinib, ruxolitinib, tofacitinib, and upadacitinib. Plasma samples are subjected to protein precipitation with MeOH, using stable isotopically labelled internal standards. The separation of JAKi in supernatants diluted 1:1 with ultrapure HO was performed using a C column Xselect HSS T3 2.5 µm, 2.1x150 mm using a mobile phase composed of formic acid (FA) 0.2% and acetonitrile (+FA 0.1%) in gradient mode. The analytical run time for the multiplex assay was 7 min. JAKi drugs were monitored by electrospray ionization in the positive mode followed by triple-stage quadrupole MS/MS analysis. The method was validated according to SFSTP and ICH guidelines over the clinically relevant concentration ranges (0.5-200 ng/mL for abrocitinib, baricitinib and upadacitinib; 1-400 ng/mL for tofacitinib; 0.5-400 ng/mL for ruxolitinib, and 10-800 ng/mL for fedratinib). This multiplex HPLC-MS/MS assay achieved good performances in term of trueness (91.1-113.5%), repeatability (3.0-9.9%), and intermediate precision (4.5-11.3%). We developed and validated a highly sensitive method for the multiplex quantification of the JAKi abrocitinib, baricitinib, fedratinib, ruxolitinib, tofacitinib, and upadacitinib in human plasma. The method will be applied for prospective clinical pharmacokinetic studies to determine whether TDM programs for JAKi based on residual drug concentrations can be recommended using disease-specific therapeutic ranges.
Janus 激酶抑制剂 (JAKi) 是一类用于治疗广泛自身免疫和骨髓增生性疾病的口服小分子药物。由于口服治疗的顺应性和主要由细胞色素 P450 酶驱动的代谢存在波动，JAKi 表现出显著的个体内和个体间药代动力学变异性。JAKi 具有剂量反应关系和狭窄的治疗指数：治疗药物监测 (TDM) 有望优化和调整其剂量方案，以解决与剂量和安全性相关的疗效和耐受性问题。本研究开发并验证了一种使用多重高效液相色谱-串联质谱 (HPLC-MS/MS) 的灵敏分析方法，用于同时定量测定目前使用的 6 种主要 JAKi，即阿布昔替尼、巴瑞替尼、非达替尼、芦可替尼、托法替尼和培哚替尼。使用甲醇进行蛋白沉淀，并用稳定同位素标记的内标进行血浆样品预处理。在超纯水中将上清液稀释 1:1 后，使用 C 柱 Xselect HSS T3 2.5 µm，2.1x150 mm，以包含 0.2%甲酸 (FA) 和乙腈 (+FA 0.1%) 的梯度模式进行分离。多重分析的分析运行时间为 7 分钟。通过电喷雾电离在正模式下监测 JAKi 药物，随后进行三重四极杆 MS/MS 分析。该方法根据 SFSTP 和 ICH 指南进行验证，涵盖了临床相关浓度范围（0.5-200ng/mL 用于阿布昔替尼、巴瑞替尼和培哚替尼；1-400ng/mL 用于托法替尼；0.5-400ng/mL 用于芦可替尼；10-800ng/mL 用于非达替尼）。该多重 HPLC-MS/MS 测定法在准确性（91.1%-113.5%）、重复性（3.0%-9.9%）和中间精密度（4.5%-11.3%）方面表现出良好的性能。我们开发并验证了一种用于人血浆中 JAKi 阿布昔替尼、巴瑞替尼、非达替尼、芦可替尼、托法替尼和培哚替尼的多重定量分析的高灵敏度方法。该方法将应用于前瞻性临床药代动力学研究，以确定基于残留药物浓度的 JAKi 的 TDM 方案是否可以使用疾病特异性治疗范围推荐。
