基于超高效液相色谱-串联质谱快速检测索非布韦及其代谢物在大鼠体内的生物等效性分析

Evaluation of a rapid UPLC-MS/MS method for determination of sofosbuvir and its metabolite in rat plasma:application to a bioequivalence study

目的:建立超高效液相色谱-串联质谱(UPLC-MS/MS)快速并同时测定大鼠血浆中抗丙肝药索非布韦及其代谢物GS-331007的含量,探讨索非布韦代谢产物作为标记物测定药时曲线的可能性,研究不同厂家抗丙肝药索非布韦在大鼠体内的生物等效性。方法:通过液质联用检测原研药A和仿制药B以36 mg·kg-1灌胃大鼠各时间点索非布韦和GS-331007的血药浓度。用DAS2.1.1和SPSS17.0软件计算药动学参数并比较原研药A和仿制药B的一致性。结果:原研药A和仿制药B中索非布韦药动学参数Cmax分别为(1 376.08±174.95)ng·mL^(-1)和(1 297.58±164.93)ng·mL^(-1),t_(max)分别为(0.75±0.08)h和(0.72±0.16)h,t1/2分别为(1.57±0.20)h和(1.73±0.45)h,AUC(0→t)分别为(2 691.67±280.85)ng·mL^(-1)·h和(2 851.20±199.54)ng·mL^(-1)·h,AUC_(0→∞)分别为(2 748.51±258.91)ng·mL^(-1)·h和(3 007.75±364.02)ng·mL^(-1)·h,原研药A和仿制药B代谢物GS-331007 Cmax分别为(1 302.52±163.73)ng·mL^(-1)和(1 430.88±107.52)ng·mL^(-1),t_(max)分别为(3.97±0.74)h和(3.95±1.38)h,t1/2分别为(5.56±2.55)h和(5.44±1.38)h,AUC(0→t)分别为(9 723.24±1170.38)ng·mL^(-1)·h和(9 032.31±1 037.76)ng·mL^(-1)·h,AUC_(0→∞)分别为(9 893.26±1 251.89)和(9 316.90±1 293.44)ng·mL^(-1)·h。结论:本实验建立的UPLC-MS/MS方法可在3.5 min内准确测定大鼠血浆中索非布韦及其代谢物GS-331007含量。根据索非布韦和其代谢物GS-331007药时曲线得出原研药A和仿制药B的药动学参数一致性较好(P>0.05)。本工作发现用代谢物GS-331007作为索非布韦生物等效性研究的可能性。

OBJECTIVE To establish a rapid and simultaneous determination method of sofosbuvir,an anti-hepatitis C virus（HCV）drug,and its metabolite GS-331007 in rat plasma by high performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）,investigate sofosbuvir metabolites as markers for determinations of drug concentration time curve and explore the bioequivalence of sofosbuvir produced by different manufacturers in rats.METHODS The plasma concentrations of sofbuvir and GS-331007 at different time points were measured by liquid chromatography and mass spectrometry in rats at 36 mg/kg.The DAS 2.1.1 and SPSS 17.0 software were used to calculate the pharmacokinetic parameters and to compare the identity of original drug A and generic drug B.RESULTS For the original drug A and generic drug B of sofosbuvir,pharmacokinetic parameters of Cmax were（1376.08±174.95）and（1297.58±164.93）ng·mL-1,tmax（0.75±0.08）and（0.72±0.16）h,t1/2（1.57±0.20）and（1.73±0.45）h,AUC（0→t）were（2691.67±280.85）and（2851.20±199.54）ng·mL-1·h,AUC（0→∞）（2748.51±258.91）and（3007.75±364.02）ng·mL-1·h,respectively.The Cmaxof metabolite GS-331007 in original drug A and generic B were（1302.52±163.73）and（1430.88±107.52）ng·mL-1,tmax（3.37±0.74）and（3.25±1.38）h,t1/2（5.56±2.55）and（5.44±1.38）h,AUC（0→t）（9723.24±1170.38）and（9032.31±1037.76）ng·mL-1·h,AUC（0→∞）（9893.26±1251.89）and（9316.90±1293.44）ng·mL-1·h.CONCLUSION The UPLC-MS/MS method established in this experiment can accurately determine the contents of sofosbuvir and its metabolite GS-331007 in rat plasma within 3.5 minutes.Based on the drug-time curve of sofobuvir and its metabolite GS-331007,the pharmacokinetic parameters of original drug A and generic drug B are consistent（P〈0.05）.This study finds out the possibility of using the metabolite GS-331007 as a bioavailability study of sofosbuvir.