Sunovion Pharmaceuticals Inc., 84 Waterford Drive, Marlborough, MA, 01752, United States.
Sunovion Pharmaceuticals Inc., 84 Waterford Drive, Marlborough, MA, 01752, United States.
J Pharm Biomed Anal. 2020 Oct 25;190:113493. doi: 10.1016/j.jpba.2020.113493. Epub 2020 Jul 26.
A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the simultaneous quantification of apomorphine and its metabolites apomorphine sulfate and norapomorphine in human plasma for supporting clinical development of a novel apomorphine sublingual thin film (APL) for the treatment of Parkinson's disease. Analytes and internal standards (IS) were extracted from human plasma by Oasis HLB SPE cartridge, followed by a reversed phase LC-MS/MS analysis using multiple reaction monitoring (MRM) in positive mode (m/z 268 → 237 for apomorphine, 348 → 237 for apomorphine sulfate, and 348 → 237 for norapomorphine). Stable isotope-labeled compounds were used as IS for respective analytes. The validated curve ranges were 0.02-20 ng/mL, 10-1000 ng/mL, and 0.5-20 ng/mL for apomorphine, apomorphine sulfate and norapomorphine, respectively. Extraction recoveries were found to be 73.4 % (apomorphine), 81.1 % (apomorphine sulfate), and 58.6 % (norapomorphine). Established long-term plasma frozen storage stabilities were 504 days at -20 °C and276 days at -60 °C, respectively. The method has been successfully used for analyzing pharmacokinetics (PK) samples collected from a comparative bioavailability study of APL and the marketed apomorphine subcutaneous (s.c.) product Apo-go®. The results demonstrated that the 15-mg APL film administrated via sublingual produced comparable PK characteristics of apomorphine when compared to the commercial product Apo-go (2-mg) via s.c. administration, hence establishing the dose regimen for this sublingual formulation. It was also noticed that the sublingual 15-mg APL film produced a significantly higher apomorphine sulfate metabolite level than the 2-mg s.c. Apo-go, and both treatments yielded a negligible level of norapomorphine metabolite in humans.
建立并验证了一种液相色谱-串联质谱（LC-MS/MS）方法，用于同时定量人血浆中的阿扑吗啡及其代谢物硫酸阿扑吗啡和去甲阿扑吗啡，以支持新型阿扑吗啡舌下薄膜（APL）治疗帕金森病的临床开发。分析物和内标（IS）用人血浆 Oasis HLB SPE 小柱提取，然后采用正相多反应监测（MRM）模式进行反相 LC-MS/MS 分析（m/z 268→237 用于阿扑吗啡，348→237 用于硫酸阿扑吗啡，348→237 用于去甲阿扑吗啡）。稳定同位素标记化合物被用作各自分析物的 IS。阿扑吗啡、硫酸阿扑吗啡和去甲阿扑吗啡的验证曲线范围分别为 0.02-20 ng/mL、10-1000 ng/mL 和 0.5-20 ng/mL。提取回收率分别为 73.4%（阿扑吗啡）、81.1%（硫酸阿扑吗啡）和 58.6%（去甲阿扑吗啡）。分别在-20°C 和-60°C 下，长期血浆冷冻保存稳定性可达到 504 天和 276 天。该方法已成功用于分析 APL 与市售阿扑吗啡皮下（s.c.）产品 Apo-go®比较生物利用度研究中采集的药代动力学（PK）样品。结果表明，与商业产品 Apo-go（2-mg）通过 s.c.给药相比，通过舌下给予 15-mg APL 薄膜产生了可比的阿扑吗啡 PK 特征，从而为这种舌下制剂建立了剂量方案。还注意到，舌下 15-mg APL 薄膜产生的硫酸阿扑吗啡代谢物水平明显高于 2-mg s.c. Apo-go，两种治疗方法在人体中均产生可忽略不计的去甲阿扑吗啡代谢物水平。
