Inhibitory effect of active ingredients of Tripterygium wilfordii Hook.F.on human carboxylesterases  

雷公藤有效成分对人羧酸酯酶的抑制作用

作　　者：LIANG Jiahong GONG Jiamin DU Zuo 梁佳鸿;龚家敏;杜佐(川北医学院公共卫生学院,四川南充637100;川北医学院临床医学院,四川南充637100)

机构地区：[1]School of Public Health,North Sichuan Medical College,Nanchong 637100,China [2]School of Clinical Medicine,North Sichuan Medical College,Nanchong 637100,China

出　　处：《中国药理学与毒理学杂志》2024年第9期652-660,共9页Chinese Journal of Pharmacology and Toxicology

基　　金：川北医学院博士科研启动基金(CBY20-QD02);南充市市校科技战略合作专项资金项目(20SXQT0318)。

摘　　要：OBJECTIVE The inhibitory effect of active ingredients of Tripterygium wilfordii Hook.F.(TWHF)(celastrol,triptolide,triptonide,wilforlide A,wilforgine and wilforine)on human carboxylester⁃ase 1(CES1)and CES2 was detected to investigate the herb-drug interactions(HDIs)of TWHF.METHODS Human liver microsomes catalysed hydrolysis of 2-(2-benzoyl-3-methoxyphenyl)benzothi⁃azole(BMBT)and fluorescein diacetate(FD)were used as the probe reaction to phenotype the activity of CES1 and CES2,respectively.The residual activities of CES1 and CES2 were detected by ultrahigh performance liquid chromatography(UPLC)after intervention with celastrol,triptolide,triptonide,wilforlide A,wilforgine and wilforine(100μmol·L^(-1)).Kinetics analysis,involving half inhibitory concentra⁃tion(IC_(50)),inhibition type and kinetic parameter(Ki),and in vitro-in vivo extrapolation(IVIVE),was carried out to predict the HDIs between these compounds and CES-metabolizing drugs.Molecular docking was performed to analyze the ligand-enzyme interaction.RESULTS Out of the six main con⁃stituents of TWHF,only celastrol exhibited strong inhibition towards both CES1 and CES2,with the inhibitory rates of 97.45%(P<0.05)and 95.62%(P<0.05),respectively.The IC_(50)was 9.95 and 4.02 mol·L^(-1),respectively,and the types of inhibition were all non-competitive inhibition.Based on the kinetics analysis,the Ki values were calculated to be 5.10 and 10.55μmol·L^(-1)for the inhibition of celastrol on CES1 and CES2,respectively.IVIVE indicated that celastrol might disturb the metabolic hydrolysis of clinical drugs in vivo by inhibiting CES1.Molecular docking results showed that hydrogen bonds and hydrophobic contacts contributed to the interaction of celastrol and CESs.CONCLUSION The inhibitory effect of celastrol on CES1 and CES2 might cause HDIs with clinical drugs hydrolysed by CESs.目的通过检测雷公藤有效成分(雷公藤红素、雷公藤甲素、雷公藤内酯酮、雷公藤内酯甲、雷公藤晋碱和雷公藤次碱)对人羧酸酯酶1(CES1)和CES2的抑制作用研究雷公藤药物-药物相互作用(HDI)机制。方法以2-(2-苯甲酰基-3-甲氧基苯基)苯并噻唑(BMBT)和荧光素二乙酸酯(FD)分别作为CES1和CES2的特异性底物,在体外构建人肝微粒体催化反应孵育体系,采用超高效液相色谱(UPLC)法分别检测雷公藤红素、雷公藤甲素、雷公藤内酯酮、雷公藤内酯甲、雷公藤晋碱和雷公藤次碱(100μmol·L^(-1))对CES1和CES2的抑制作用,包括抑制率以及半抑制浓度(IC_(50))、抑制类型和抑制动力学常数(Ki)等动力学参数。通过体外-体内外推法(IVIVE)预测雷公藤有效成分与CES代谢药物之间的HDI,并通过分子对接研究其分子间相互作用。结果雷公藤的6个主要成分中,只有雷公藤红素100μmol·L^(-1)对CES1和CES2均表现出强抑制作用,抑制率分别为97.45%(P<0.05)和95.62%(P<0.05),IC_(50)分别为9.95和4.02μmol·L^(-1),抑制类型均为非竞争性抑制,Ki分别为5.10和10.55μmol·L^(-1)。IVIVE结果表明,雷公藤红素可能在体内通过抑制CES1产生HDI。分子对接结果显示,雷公藤红素和CES通过氢键和疏水作用产生分子间相互作用。结论雷公藤红素对CES1和CES2具强烈抑制作用,可能在体内通过影响CES相关药物的代谢产生HDI。

关 键 词：Tripterygium wilfordii Hook.F. CELASTROL CARBOXYLESTERASES enzyme inhibition 

分 类 号：R96[医药卫生—药理学] R285[医药卫生—药学]