Zhong Bo, Li Yaxin, Idippily Nethrie, Petty Aaron, Su Bin, Wang Bingcheng
Department of Chemistry, Center for Gene Regulation in Health and Disease, College of Sciences and Health Professions, Cleveland State University, Cleveland, Ohio, USA.
Rammelkamp Center for Research and Department of Medicine, MetroHealth System, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA.
Biomed Chromatogr. 2019 Apr;33(4):e4461. doi: 10.1002/bmc.4461. Epub 2019 Jan 16.
Compound 27 {1, 12-bis[4-(4-amino-6,7-dimethoxyquinazolin-2-yl)piperazin-1-yl]dodecane-1,12-dione} is a novel small molecule agonist of EphA2 receptor tyrosine kinase. It showed much improved activity for the activation of EphA2 receptor compared with the parental compound doxazosin. To support further pharmacological and toxicological studies of the compound, a method using liquid chromatography and electrospray ionization tandem mass spectrometry (LC-MS/MS) has been developed for the quantification of this compound. Liquid-liquid extraction was used to extract the compound from mouse plasma and brain tissue homogenate. Reverse-phase chromatography with gradient elution was performed to separate compound 27 from the endogenous molecules in the matrix, followed by MS detection using positive ion multiple reaction monitoring mode. Multiple reaction monitoring transitions m/z 387.3 → 290.1 and m/z 384.1 → 247.1 were selected for monitoring compound 27 and internal standard prazosin, respectively. The linear calibration range was 2-200 ng/mL with the intra- and inter-day precision and accuracy within the acceptable range. This method was successfully applied to the quantitative analysis of compound 27 in mouse plasma and brain tissue with different drug administration routes.
化合物27{1,12 - 双[4 - (4 - 氨基 - 6,7 - 二甲氧基喹唑啉 - 2 - 基)哌嗪 - 1 - 基]十二烷 - 1,12 - 二酮}是一种新型的EphA2受体酪氨酸激酶小分子激动剂。与母体化合物多沙唑嗪相比，它对EphA2受体激活表现出显著提高的活性。为支持该化合物进一步的药理和毒理学研究，已开发出一种使用液相色谱和电喷雾电离串联质谱(LC - MS/MS)的方法来定量该化合物。液 - 液萃取用于从小鼠血浆和脑组织匀浆中提取该化合物。采用梯度洗脱的反相色谱法将化合物27与基质中的内源性分子分离，随后使用正离子多反应监测模式进行质谱检测。分别选择多反应监测跃迁m/z 387.3→290.1和m/z 384.1→247.1来监测化合物27和内标哌唑嗪。线性校准范围为2 - 200 ng/mL，日内和日间精密度及准确度均在可接受范围内。该方法已成功应用于不同给药途径下小鼠血浆和脑组织中化合物27的定量分析。
