Analytical and Bioanalytical Development, Bristol-Myers Squibb Company, Princeton, NJ 08543, USA.
J Pharm Biomed Anal. 2013 Sep;83:237-48. doi: 10.1016/j.jpba.2013.05.019. Epub 2013 May 20.
BMS-927711 is a calcitonin gene-related peptide (CGRP) receptor antagonist that is being developed for the treatment of migraine. A rapid, accurate and robust assay was developed and validated for the quantitation of BMS-927711 in rat, monkey, rabbit and mouse plasma using ultra high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS). A simplified method screening strategy was utilized that included a liquid-liquid extraction (LLE) methodology and eleven LC columns (ten sub-2 μm UHPLC columns and one 2.6 μm HPLC column) for screening with emphasis on the removal of phospholipids, avoidance of metabolite interference and ruggedness of LC conditions. A stable isotope labeled [(13)C2, D4]-BMS-927711 was used as the internal standard, and 50 μL of plasma samples were used for extraction by automated LLE with methyl tert-butyl ether (MTBE) in 96-well format. Chromatographic separation was achieved with an isocratic elution and a gradient column wash on a Waters Acuity UPLC(®) BEH C18 column (2.1 mm × 50 mm, 1.7 μm) with run time of 3.7 min. Positive electrospray ionization was performed using selected reaction monitoring (SRM) with transitions of m/z 535>256 for BMS-927711 and m/z 541>256 for [(13)C2, D4]-BMS-927711. The standard curve, which ranged from 3.00 to 3000 ng/mL for BMS-927711, was fitted to a 1/x(2) weighted linear regression model. The intra-assay precision was within 5.2% CV, inter-assay precision was within 5.9% CV, and the assay accuracy was within ±5.2% deviation (%Dev) of the nominal values in all the species. The stability of an N-carbamoyl glucuronide metabolite was carefully investigated, and the conversion of this metabolite to BMS-927711 was minimal and manageable without a stabilization procedure. The method was successfully applied to multiple non-clinical toxicokinetic studies in different species in support of the investigative new drug (IND) filing.
BMS-927711 是一种降钙素基因相关肽 (CGRP) 受体拮抗剂，目前正在开发用于治疗偏头痛。本研究建立并验证了一种用于定量检测大鼠、猴子、兔子和小鼠血浆中 BMS-927711 的超高效液相色谱-串联质谱法（UHPLC-MS/MS）。本研究采用简化的方法筛选策略，包括液液萃取（LLE）方法和 11 根 LC 柱（10 根亚 2μm UHPLC 柱和 1 根 2.6μm HPLC 柱）进行筛选，重点在于去除磷脂、避免代谢物干扰和 LC 条件的稳健性。以稳定同位素标记的 [(13)C2, D4]-BMS-927711 作为内标，采用自动 LLE 用甲基叔丁基醚（MTBE）在 96 孔板中提取 50μL 血浆样品。色谱分离采用等度洗脱和梯度柱冲洗在 Waters Acuity UPLC®BEH C18 柱（2.1mm×50mm，1.7μm）上实现，运行时间为 3.7min。正电喷雾电离采用选择反应监测（SRM）进行，BMS-927711 的母离子-子离子跃迁为 m/z 535>256，[(13)C2, D4]-BMS-927711 的母离子-子离子跃迁为 m/z 541>256。BMS-927711 的标准曲线范围为 3.00-3000ng/mL，拟合为 1/x(2)加权线性回归模型。在所有物种中，内、日间精密度均小于 5.2%CV，准确度均在名义值的±5.2%偏差（%Dev）范围内。仔细研究了 N- 氨甲酰葡萄糖醛酸代谢物的稳定性，该代谢物转化为 BMS-927711 的转化率很小且可以控制，无需稳定化处理。该方法成功应用于支持 investigational new drug（IND）申报的多个不同物种的非临床毒代动力学研究。
