艾可清对药物代谢酶活性及对洛匹那韦与利托那韦的大鼠血浆药代动力学参数的影响  

Effects of Aikeqing on Drug-Metabolizing Enzyme Activity and on Pharmacokinetic Parameters of Lopinavir and Ritonavir in Rat Plasma

作　　者：何晓峰 卢元媛[2] 陈剑涛[2] 符林春[2] 沈小玲[1,2] 胡英杰[1,2] HE Xiaofeng;LU Yuanyuan;CHEN Jiantao;FU Linchun;SHEN Xiaoling;HU Yingjie(Science and Technology Innovation Center,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Institute of Tropical Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学科技创新中心,广东广州510405 [2]广州中医药大学热带医学研究所,广东广州510405

出　　处：《中药新药与临床药理》2024年第7期1046-1054,共9页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家科技重大专项课题(2014ZX10005002)。

摘　　要：目的考察中药复方艾可清对药物代谢酶和抗艾滋病病毒(HIV)药物克力芝组分洛匹那韦(LPV)和利托那韦(RTV)药物代谢动力学的影响。方法将人肝微粒体与混合探针和艾可清共孵育,用高效液相色谱-质谱(HPLC-MS)联用技术测定探针的含量变化,计算艾可清对肝微粒体中药物代谢酶细胞色素P450不同亚型的半数抑制浓度(IC50),确定艾可清抑制的P450亚型。建立HPLC-MS法同时测定大鼠血浆中LPV和RTV含量;大鼠灌胃艾可清或溶媒,每日1次,连续7 d,末次灌胃艾可清后0.5 h灌胃克力芝,检测LPV和RTV的血药浓度,分析艾可清对其药代动力学参数的影响。结果艾可清甲醇提取物在0~500μg·mL^(-1)浓度范围内,对P450代谢酶表型CYP2D6、CYP2C8、CYP2E1、CYP2C19、CYP1A2、CYP2B6、CYP2C9和CYP3A4酶活性的IC50值依次为7.7、75.3、144.0、99.5、43.5、104.5、49.3和204.9μg·mL^(-1)。建立了定量分析LPV和RTV的HPLC-MS/MS方法,LPV和RTV分别在30~10000 ng·mL^(-1)和3~1000 ng·mL^(-1)范围呈线性关系,最低定量限分别为30 ng·mL^(-1)和3 ng·mL^(-1),日内、日间精密度均小于5%,LPV和RTV的准确度均在96.3%~109.0%之间,提取回收率均不小于88.7%,基质效应均在93.8%~105.0%之间,血浆样品稳定性较好。与克力芝组比较,艾可清与克力芝联用组LPV的非房室模型参数AUC0-t、AUC0-∞、MRT0-t、t1/2z、tmax、Vz/F、Clz/F、Cmax均无明显影响(P>0.05),但可延长MRT0-∞(P<0.05);联用组RTV所有药动学参数均无明显影响(P>0.05)。结论艾可清对人肝脏药物代谢酶CYP450中8个亚型有不同程度的抑制作用,其中对CYP2D6的抑制作用最明显;大鼠灌胃人等效剂量艾可清对洛匹那韦和利托那韦的药代动力学参数无明显的影响。Objective To investigate the effects of Aikeqing(AKQ),a compound of Chinese medicine,on drugmetabolizing activity and on the pharmacokinetic parameters of HIV-1 protease inhibitors lopinavir(LPV)and ritonavir(RTV)in Kaletra tablets.Methods Human liver microsomes were co-incubated with mixed probes andAKQ.HPLC-MS was employed to measure the content of probe.Half-inhibitory concentration(IC50)of AKQ on different subtypes of cytochrome P450 enzymes involved in drug metabolism of liver microsomes was calculated.The P450 subtype,whose activity was significantly inhibited by AKQ was then identified.HPLC-MS analytical method for simultaneous determination of LPV and RTV in rat plasma was established.SD rats were orally given AKQ or vehicle once a day for 7 consecutive days.After half an hour of the last gavage of AKQ,the rats were given Kaletra by intragastric administration.Then,the blood concentration of LPV and RTV were measured and the effect of AKQ on pharmacokinetic parameters of LPV and RTV were analyzed.Results The methanol extract of AKQ at the concentrations of 0~500μg·mL^(-1)showed inhibitory effects on the metabolic activity of CYP2D6,CYP2C8、CYP2E1,CYP2C19,CYP1A2,CYP2B6,CYP2C9 and CYP3A4,with IC50 values of 7.7,75.3,144.0,99.5,43.5,104.5,49.3 and 204.9μg·mL^(-1),respectively.An HPLC-MS/MS method was established for simultaneous quantification of LPV and RTV in blood samples.LPV and RTV showed linear relationships in the ranges of 30~10000 ng·mL^(-1)and 3~1000 ng·mL^(-1),respectively.The lowest limits of quantification were 30 ng·mL^(-1)and 3 ng·mL^(-1).Intra-day and inter-day precision were all less than 5%,and the accuracy of LPV and RTV was in the range of 96.3%~109%.The extraction recovery rates were not less than 88.7%,the matrix effects were 93.8%~105.0%,and the plasma samples were stable.Compared with Kaletra group,there was no significant changes in noncompartmental model parameters including AUC0-t,AUC0-∞,MRT0-t,t1/2z,tmax,Vz/F,Clz/F and Cmax of LPV in AKQ+Kaletra group(P>0.05).But MRT0-�

关 键 词：艾可清 细胞色素P450 洛匹那韦 利托那韦 高效液相色谱-质谱 药代动力学 大鼠 

分 类 号：R285.5[医药卫生—中药学]