N-苄氧羰基-甘脯酰表阿霉素体内分析方法的建立与血药浓度测定

目的: 建立一种灵敏、稳定的新型靶向侯选化合物N-苄氧羰基-甘氨酰脯氨酸表阿霉素(Z-GP-EPI)体内分析方法,并考察大鼠单次给药血药浓度变化及求算药代动力学参数。 方法: 液-液萃取法进行样品处理。色谱条件:色谱柱为Ultimate^®XB-C₁₈(4.6 mm×250 mm,5 μm);流动相为乙腈-0.1% TFA水(45∶55,v/v);流速为1 mL·min^-1;荧光检测器,激发波长为495 nm,发射波长为590 nm。SD大鼠颈静脉注射给药(18 mg·kg^-1),HPLC测定血药浓度经时变化,绘制平均血药浓度-时间曲线,统计矩模型估算药代动力学参数。 结果: Z-GP-EPI在0.1~100 μg·mL^-1浓度范围内线性相关性良好(r=0.9993),回归方程为:Y=0.526X-0.014,定量下限为0.1 μg·mL^-1。低、中、高质控样品的萃取回收率、日间/日内精密度与准确度均符合化学药物非临床药代动力学技术指导原则。主要药代动力学参数如下:AUC_0-∞为(10.165±2.863)μg·mL^-1·h,MRT_0-∞为(1.283±0.629)h,t_1/2z为(2.208±1.475)h,V_z为(5.458±2.80)L·kg^-1,CL_z为(1.714±0.885)L·h^-1·kg。 结论: 建立稳定、可靠的Z-GP-EPI体内分析方法,为后续开展系统性药代动力学研究奠定基础。

Objective: To develop a sensitive and stable in vivo analysis method for detecting concentration of new targeted candidate antitumor prodrug Z-GP-EPI.The changes of blood concentration after single administration to rats and pharmacokinetic parameters will be investigated. Methods: Liquid-liquid extraction was used for sample preparation.Chromatographic separation was performed on an UltimateXB C₁₈ column(4.6 mm×250 mm,5 μm)with the mobile phase of acetonitrile-0.1% trifluoroacetic acid water(45∶55,v/v)at a flow rate of 1 mL·min^-1.The detector was fluorescence detection under the excitation wavelength of 495 nm and the emission wavelength of 590 nm.The variation of concentration-time was detected by HPLC,then the mean concentration-time profile was described and the pharmacokinetic parameters was calculated with statistical moment after administration via jugular vein at a dose of 18 mg·kg^-1 to Sprague-Dawley rats. Results: A good linearity was observed over the concentration range of 0.1-100 μg·mL^-1 and the correlation coefficient was 0.9993.The regression equation was Y=0.526X-0.014 and the limit of quantity was 0.1 μg·mL^-1.The extraction recovery,intra-and inter-day precision and accuracy of low,medium,high quality controlled samples were qualified according to the technological guiding principle of non-clinical pharmacokinetics of chemical drugs.The main pharmacokinetic parameters were measured:AUC_0-∞was(10.165±2.863)μg·mL^-1·h,MRT_0-∞was(1.283±0.629)h,t_1/2z was(2.208±1.475)h,V_z was(5.458±2.80)L·kg^-1,CL_z was(1.714±0.885)L·h^-1·kg. Conclusion: A stable and reliable method that could be used for further systematic pharmacokinetic study is established.