南北五味子提取物对大鼠体内环磷酰胺代谢的影响

目的:探讨南北五味子提取物分别对环磷酰胺(CTX)及其代谢产物羧基磷酰胺(CPM)、4-酮基环磷酰胺(4-Keto CTX)、脱氯乙基环磷酰胺(DC-CTX)血浆药动学及尿排泄的影响,从药物相互作用影响环磷酰胺代谢的角度阐明南北五味子的减毒作用机制。方法:采用UHPLC-MS/MS法同时测定大鼠血浆或尿液中CTX及其代谢产物的浓度。将SD大鼠随机分组分别给药分别取样,采集不同时间点血浆样品,绘制药-时曲线,并用DAS2.0软件一室模型拟合药动学参数,使用单因素方差分析法比较组间药动学参数的差异性;采集不同时间段尿液样本,计算待测成分的累积排泄量。结果:CTX单独给药组、合并南五味子提取物(制剂五酯胶囊,WZC,300 mg·kg^-1)给药组,以及合并北五味子醇提物(BWWZ,750 mg·kg^-1)给药组,不同待测物的AUC(μg·h^-1·mL^-1)分别为:CTX:411.818±82.10,483.492±174.49,683.92±91.70;DC-CTX:683.97±91.70,35.95±70.51,8.44±3.96;4-Keto CTX:87.27±38.81,25.78±10.03,17.18±37.2;CPM:99.74±16.18,110.21±57.99,104.90±16.23。Cmax(μg·mL^-1)分别为:CTX:233.41±32.10,255.021±45.90,249.27±17.52;DC-CTX:3.63±1.25,1.21±13.48,1.31±0.72;4-Keto CTX:5.24±1.04,5.34±2.41,2.84±0.81;CPM:25.88±5.26,27.53±8.11,22.42±4.75。48 h内的累积排泄量(μg)分别为:CTX:1 455.36,3 829.16,4 830.69;DC-CTX:2 136.21,921.18,468.87;4-Keto CTX:759.18,1 348.54,338.38;CPM:9 268.06,17 386.18,8 902.05。结论:WZC和BWWZ对CTX的代谢产物DC-CTX的药动学及尿液排泄均有显著影响,均可通过抑制CYP3A4活性使得侧链代谢毒性产物氯乙醛的生成明显降低;结合血浆、尿液中的实验结果可知在血浆中浓度较高的待测物在尿液中排泄也较多,使用WZC和BWWZ均可促进CTX原型和CPM的排泄。

Effects of Wuzhi capsules and Schisandrae chinensis fructus extract on cyclophosphamide metabolism in rats

OBJECTIVE To discuss the effects of Wuzhi capsules (WZC) and Schisandrae chinensis fructus extract (BWWZ) on the pharmacokinetics and urinary excretion behaviors of cyclophosphamide (CTX) as well as its metabolites carboxyl phosphoramide (CPM), 4-Keto cyclophosphamide (4-Keto CTX) as well as dechlorination ethyl cyclophosphamide (DC-CTX), and to illustrate the mechanism of toxicity attenuation effects of WZC and BWWZ by exploring their interactions with CTX metabolism.METHODS A valid UHPLC-MS/MS method was applied for simultaneous determination of CTX as well as its metabolites in rat plasma and urine. The Sprague Dawley rats were randomly divided into three groups (CTX group; BWWZ group, 750 mg·kg^-1; WZC group, 300 mg·kg^-1) and plasma samples were collected at different time points using an orbital sampling method. The concentration time curve was constructed using a DAS2.0 software for calculation of the pharmacokinetic parameters by one-compartment model. The pharmacokinetic parameters from different groups were compared by one-way ANOVA for significance analysis. In addition, urine samples were gained from different time periods to obtain the total cumulative excretion.RESULTS The areas under the curve (AUC,μg·h·mL^-1) of the analytes were 411.818±82.10, 483.492±174.49 and 683.92±91.70 for CTX; 683.97±91.70, 35.95±70.51 and 8.44±3.96 for DC-CTX; 87.27±38.81, 25.78±10.03 and 17.18±37.2 for 4-Keto CTX; 99.74±16.18, 110.21±57.99 and 104.90±16.23 for CPM in CTX, WZC and BWWZ groups, respectively. Maximum plasma concentrations (C_max,μg·mL^-1) of the analytes were 233.41±32.10, 255.021±45.90 and 249.27±17.52 for CTX; 3.63±1.25, 1.21±13.48 and 1.31±0.72 for DC-CTX; 5.24±1.04, 5.34±2.41 and 2.84±0.81 for 4-Keto CTX; 25.88±5.26, 27.53±8.11 and 22.42±4.75 for CPM in CTX, WZC and BWWZ groups. The 48 h urinary cumulative excretions(μg) were 1 455.36, 3 829.16 and 4 830.69 for CTX; 2 136.21, 921.18 and 468.87 for DC-CTX; 759.18, 1 348.54 and 338.38 for 4-Keto CTX; 9 268.06, 17 386.18 and 8 902.05 for CPM in CTX, WZC and BWWZ groups.CONCLUSION Both WZC and BWWZ can influence the pharmacokinetic behaviors of DC-CTX which results in the reduction of the toxic metabolite chloroacetaldehyde, as a consequence of CYP3A4 inhibition. It can be observed from the experiment that the rich abundance of the analyte in plasma, the greater the urinary excretion. Both WZC and BWWZ can accelerate the excretion of CTX and CPM.