Department of Chemistry, Cleveland State University, 2121 Euclid Avenue, Cleveland, OH 44115, United States.
Case Comprehensive Cancer Center, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, United States.
J Pharm Biomed Anal. 2018 May 10;153:145-151. doi: 10.1016/j.jpba.2018.02.034. Epub 2018 Feb 19.
GS87 is a novel, highly specific GSK3 inhibitor, which has shown to induce extensive differentiation of acute myeloid leukemia (AML) cells in early mouse studies and has great potential for therapeutic advancement. This work described the development and validation of an LC-MS/MS method for quantitative determination of GS87 in mouse plasma. In this method, GS87 and T6447952 (a structural analog used as internal standard) were extracted from plasma using hexane as extraction solvent, and separated isocratically on a Waters XTerra MS C8 column (2.1 × 50 mm, 3.5 μm) using a mobile phase consisting of acetonitrile and 5.00 mM ammonium formate (35:65, v/v) pumped at a flow rate of 0.200 mL min. Quantitation of GS87 was done by positive electrospray ionization tandem mass spectrometry operated in multiple-reaction-monitoring (MRM) mode. The method has been validated in accordance with the US Food and drug administration guidelines for bioanalytical method validation. It has linear calibration range of 2.50-250 ng mL with correlation coefficient of >0.999. The intra- and inter- assay accuracy and precision were ≤ ±5 and ≤6%, respectively. The IS normalized recovery of GS87 was 103-106%. The stability studies showed that GS87 was stable under all tested conditions. The method developed has been successfully applied to the measurement of GS87 concentrations in mouse plasma samples from an animal study, and may be useful for further preclinical investigation of GS87.
GS87 是一种新型的、高度特异的 GSK3 抑制剂，在早期的小鼠研究中已显示出能诱导广泛的急性髓性白血病（AML）细胞分化，具有很大的治疗进展潜力。本工作描述了一种用于定量测定小鼠血浆中 GS87 的 LC-MS/MS 方法的开发和验证。在该方法中，GS87 和 T6447952（用作内标的结构类似物）用正己烷作为提取溶剂从血浆中提取，然后在 Waters XTerra MS C8 柱（2.1×50mm，3.5μm）上以等度洗脱，流动相由乙腈和 5.00mM 甲酸铵（35:65，v/v）组成，流速为 0.200mL/min。GS87 的定量分析采用正电喷雾串联质谱，以多反应监测（MRM）模式运行。该方法已按照美国食品和药物管理局（FDA）生物分析方法验证指南进行验证。它具有 2.50-250ng/mL 的线性校准范围，相关系数>0.999。日内和日间准确度和精密度分别≤±5%和≤6%。GS87 的 IS 归一化回收率为 103-106%。稳定性研究表明，GS87 在所有测试条件下均稳定。所开发的方法已成功应用于动物研究中测定小鼠血浆样品中的 GS87 浓度，可能有助于进一步对 GS87 进行临床前研究。
