Amylin Pharmaceuticals, LLC (a wholly-owned subsidiary of Bristol-Myers Squibb), 9625 Towne Centre Drive, San Diego, CA 92121, USA.
Amylin Pharmaceuticals, LLC (a wholly-owned subsidiary of Bristol-Myers Squibb), 9625 Towne Centre Drive, San Diego, CA 92121, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Apr 15;957:24-9. doi: 10.1016/j.jchromb.2014.02.040. Epub 2014 Mar 4.
Peptide agonists of the glucagon-like peptide-1 receptor (GLP-1R) and the cholecystokinin-1 receptor (CCK1-R) have therapeutic potential because of their marked anorexigenic and weight lowering effects. Furthermore, recent studies in rodents have shown that co-administration of these agents may prove more effective than treatment either of the peptide classes alone. To correlate the pharmacodynamic effects to the pharmacokinetics of these peptide drugs in vivo, a sensitive and robust bioanalytical method is essential. Furthermore, the simultaneous determination of both analytes in plasma samples by a single method offers obvious advantages. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is well suited to this goal through its ability to simultaneously monitor multiple analytes through selected reaction monitoring (SRM). However, it is a challenge to find appropriate conditions that allow two peptides with widely disparate physiochemical properties to be simultaneously analyzed while maintaining the necessary sensitivity for their accurate plasma concentrations. Herein, we report an on-line solid phase extraction (SPE) LC-MS/MS method for simultaneous quantification of the CCK1-R agonist AC170222 and the GLP-1R agonist AC3174 in rodent plasma. The assay has a linear range from 0.0975 to 100ng/mL, with lower limits of quantification of 0.0975ng/mL and 0.195ng/mL for AC3174 and AC170222, respectively. The intra- and inter-day precisions were below 15%. The developed LC-MS/MS method was used to simultaneously quantify AC3174 and AC170222, the results showed that the terminal plasma concentrations of AC3174 or AC170222 were comparable between groups of animals that were administered with the peptides alone (247±15pg/mL of AC3174 and 1306±48pg/mL of AC170222), or in combination (222±32pg/mL and 1136±47pg/mL of AC3174 and AC170222, respectively). These data provide information on the drug exposure to aid in assessing the combination effects of AC3174 and AC170222 on rodent metabolism.
胰高血糖素样肽-1 受体 (GLP-1R) 和胆囊收缩素-1 受体 (CCK1-R) 的肽激动剂具有治疗潜力，因为它们具有明显的厌食和降低体重的作用。此外，最近在啮齿动物中的研究表明，这些药物的联合给药可能比单独治疗任何一种肽类更有效。为了将这些肽类药物的药效学作用与体内药代动力学相关联，需要一种灵敏而强大的生物分析方法。此外，通过单一方法同时测定血浆样品中的两种分析物具有明显的优势。液相色谱-串联质谱法 (LC-MS/MS) 通过其通过选择反应监测 (SRM) 同时监测多种分析物的能力非常适合实现这一目标。然而，找到允许两种具有广泛不同理化性质的肽同时进行分析的合适条件，同时保持对其准确血浆浓度的必要灵敏度，是一项挑战。本文报道了一种在线固相萃取 (SPE) LC-MS/MS 方法，用于同时定量分析啮齿动物血浆中的 CCK1-R 激动剂 AC170222 和 GLP-1R 激动剂 AC3174。该测定法的线性范围为 0.0975 至 100ng/mL，AC3174 和 AC170222 的定量下限分别为 0.0975ng/mL 和 0.195ng/mL。日内和日间精密度均低于 15%。所开发的 LC-MS/MS 方法用于同时定量分析 AC3174 和 AC170222，结果表明，单独给予肽（AC3174 为 247±15pg/mL，AC170222 为 1306±48pg/mL）或联合给予（AC3174 为 222±32pg/mL，AC170222 为 1136±47pg/mL）时，AC3174 或 AC170222 的末端血浆浓度在动物组之间是可比的。这些数据提供了有关药物暴露的信息，有助于评估 AC3174 和 AC170222 对啮齿动物代谢的联合作用。
