基于计算机虚拟技术研究花椒毒素及其同分异构体对细胞色素P450的抑制作用差异  

Study on inhibitory effects of methoxsalen and its isomers on drug metabolic enzyme CYP450 based on computer virtual technology

作　　者：刘世豪 王丽莉[1] 刘文莉 靳超群 陈佳萍 何新[1,2] LIU Shihao;WANG Lili;LIU Wenli;JIN Chaoqun;CHEN Jiaping;HE Xin(Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]天津中医药大学,天津301617 [2]广东药科大学,广东广州510006

出　　处：《药物评价研究》2021年第1期15-24,共10页Drug Evaluation Research

基　　金：教育部创新团队发展计划资助项目(IRT14R41);广东省自然科学基金资助项目(2020A1515010156);广东省普通高校重点领域专项(2019KZDZX1006)。

摘　　要：目的研究花椒毒素及其同分异构体对药物代谢酶细胞色素P450(CYPs)的抑制差异。方法采用计算机虚拟技术研究花椒毒素及其同分异构体香柑内酯、异佛手柑内酯与6种CYPs酶(CYP1A2、CYP2D6、CYP3A4、CYP2C19、CYP2C9、CYP2A6)的相互作用;采用Amber(ff99SB)软件进行花椒毒素、香柑内酯、异佛手柑内酯与CYP2A6的动力学模拟研究;采用"Cocktail"药物探针法进行验证,分别测定这3种同分异构体对人肝微粒CYPs酶活性的影响,以及对CYP2A6与CYP1A2的半数抑制浓度(IC50)。结果计算机模拟技术结果显示,花椒毒素、香柑内酯、异佛手柑内酯对6种CYPs结合能力不同,对CYP2A6的结合能力差异明显。分子对接、分子动力学结果表明,花椒毒素与CYP2A6形成的氢键ASN297∶HD22-M∶O4是花椒毒素与CYP2A6结合过程中最重要的且稳定的氢键,香柑内酯与ASN297产生的分子间氢键稳定性低于花椒毒素体系,异佛手柑内酯在动力学中所产生的氢键均不稳定;受体活性口袋中疏水性氨基酸与口袋形状互补是促成配体受体结合的重要因素。"Cocktail"药物探针法表明,花椒毒素、香柑内酯、异佛手柑内酯对CYPs均具有不同程度的抑制作用,其中对CYP1A2具有显著抑制作用,相对剩余酶活力分别为2.67%、4.64%、13.58%,其IC50分别为1.42、2.29、5.12μmol/L;对CYP2A6的抑制作用存在差异,相对剩余酶活力分别为8.10%、38.91%、74.98%,其IC50值分别为3.89、44.03、103.30μmol/L。结论花椒毒素、香柑内酯、异佛手柑内酯对CYP1A2具有明显的抑制作用,对CYP2A6的抑制效果存在明显差异。甲氧基的取代位置与呋喃环骈合的位置对3种化合物与CYP2A6结合影响较大,其中CYP2A6活性口袋中ASN297是蛋白受体与配体结合的关键氨基酸残基。Objective To study the inhibitory effects of methoxsalen and its isomers on drug metabolizing enzyme CYP450.Methods The interaction of methoxsalen and its isomers bergapten,isobergapten and six CYPs were studied by computer simulation technology.Amber(ff99 SB)software was used to simulate the dynamics of zanthoxylin,bergamot lactone,isobergamot lactone and CYP2 A6.The results of computer simulation were verified by"Cocktail"drug probe method,and their half maximal inhibitoryconcentration(IC50)values to CYP2 A6 and CYP1 A2 were determined.Result Methoxsalen,bergapten,isobergapten had different binding to the six CYPs,among which the binding to CYP2 A6 is significantly different.The results of molecular docking and molecular dynamics showed that ASN297∶HD22-M∶O4 was the most important and stable hydrogen bond between methoxsalen and CYP2 A6,The results showed that the stability of intermolecular hydrogen bond between bergapten and ASN297 was lower than that of methoxsalen system,and the hydrogen bond of isobergamot lactone in isobergapten was unstable.The complementary of hydrophobic amino acids and pocket shape in receptor active pocket was an important factor to promote ligand receptor binding.The"Cocktail"drug probe method showed that the three isomers have different degrees of inhibition on CYPs,which had significant inhibition on CYP1 A2.The relative remaining enzyme activities were 2.67%,4.64%,and 13.58%,and their IC50 values were 1.42,2.29,and 5.12μmol/L;the inhibitory effect on CYP2 A6 was different,the relative remaining enzyme activities were 8.10%,38.91%and 74.98%,and the IC50 values were 3.89,44.03,and 103.30μmol/L.Conclusion Methoxsalen,bergapten,isobergapten have significant inhibitory effects on CYP1 A2,and there are significant differences effects on CYP2 A6.Molecular docking and molecular dynamics results show that the substitution position of the methoxy group and the position of the furan ring have a greater impact on the binding of the three compounds to CYP2 A6.ASN297 in the CYP2 A6 activ

关 键 词：花椒毒素 香柑内酯 异佛手柑内酯 同分异构体 细胞色素P450 计算机虚拟技术 “Cocktail”药物探针法 

分 类 号：R969.1[医药卫生—药理学]