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目的:本文研究抗艾滋病中药复方ZYSH对人肝微粒体CYP3A4的活性抑制和对茚地那韦体外代谢的增效作用,及其对茚地那韦在SD大鼠体内暴露量的增加。方法:中药复方ZYSH总浸膏用磷酸缓冲液(PBS)稀释后与混合人肝微粒体预孵育15 min,加入探针底物睾酮和辅酶β-NADPH孵育30 min,用LC-MS/MS定量检测6β-羟基睾酮的生成,计算ZYSH对CYP3A4的抑制率(IC50)。5 mg.mL-1和10 mg.mL-1的ZYSH在与人肝微粒体共同孵育15 min后加入茚地那韦和β-NADPH分别孵育0,15,30,60,90和120min,检测复方对茚地那韦体外代谢的影响。SD大鼠体内灌胃给予ZYSH 7 d,再经灌胃给予茚地那韦,检测茚地那韦的血药浓度,观察复方ZYSH对茚地那韦的体内暴露的影响。结果:复方ZYSH能明显抑制CYP3A4的活性,IC50为3.21 mg.mL-1。5 mg.mL-1和10 mg.mL-1的ZYSH在体外能明显抑制茚地那韦的代谢,其t1/2由47.5 min变为184和1 404 min,微粒体对茚地那韦的固有清除率由对照组的36.6 mL.min-1.kg-1分别改变为9.4和1.2 mL.min-1.kg-1。灌胃给予ZYSH 7 d后茚地那韦在SD大鼠体内AUC明显增加,为对照组的2.1倍。结论:复方ZYSH能够抑制人肝微粒体CYP3A4的活性,通过代谢性相互作用减缓茚地那韦的体外代谢,增加大鼠体内暴露量。
OBJECTIVE: To study the inhibitory effect of ZYSH on human liver microsomal CYP3A4 and its synergistic effect on in vitro metabolism of indinavir and the increase of exposure to indinavir in SD rats. Methods: The total extract of ZYSH was diluted with phosphate buffered saline (PBS) and preincubated with mixed human liver microsome for 15 min. The probe substrate was incubated with coenzyme β-NADPH for 30 min and quantified by LC-MS / MS The formation of 6β-hydroxytestosterone was detected and the inhibition rate of CYP3A4 by ZYSH (IC50) was calculated. 5 mg.mL-1 and 10 mg.mL-1 of ZYSH incubated with human liver microsomes for 15 min after adding indinavir and β-NADPH were incubated for 0, 15, 30, 60, 90 and 120 min, respectively Effect of compound on in vitro metabolism of indinavir. Sprague-Dawley rats were intragastrically given ZYSH for 7 days and then given indinavir via intragastric administration. The plasma concentration of indinavir was measured to observe the effect of compound ZYSH on in vivo exposure of indinavir. Results: Compound ZYSH could significantly inhibit the activity of CYP3A4 with the IC50 of 3.21 mg.mL-1.5 mg.mL-1 and 10 mg.mL-1 of ZYSH in vitro can significantly inhibit the metabolism of indinavir, t1 / 2 from 47.5 min to 184 and 1 404 min, the intrinsic clearance rate of microsomal to indinavir was changed from 36.6 mL.min-1.kg-1 in the control group to 9.4 and 1.2 mL.min-1.kg -1. After 7 days of intragastric administration of ZYSH, the AUC of indinavir in SD rats increased significantly, 2.1 times that of the control group. Conclusion: Compound ZYSH can inhibit the activity of CYP3A4 in human liver microsomes and reduce the in vitro metabolism of indinavir through metabolic interaction and increase the in vivo exposure of rats.