细胞色素P450酶诱导-抑制的研究方法及在药物研究中的应用

CN341206/R,ISSN10092501(5):579-58520110215收稿20110312修回国家自然科学基金项目(30701101)卜明华,女,硕士研究生,研究方向:生药学Tel:01062835641Email:[e-mail]张颖,通信作者,女,副研究员,研究方向:药代动力学Tel:01062835641Email:[e-mail]细胞色素P450酶诱导/抑制的研究方法及在药物研究中的应用卜明华1,2,张颖1,刘建勋1,刘东春21中国中医科学院西苑医院实验研究中心,北京100091;2沈阳药科大学中药学院,沈阳110016,辽宁细胞色素P450(CYP450)在外源及内源物质的生物转化过程中发挥着至关重要的作用,是肝脏中重要的药物代谢酶CYP450酶活性/含量的改变直接影响了经其代谢的药物在体内的有效量和作用时间,与药效和用药安全性有密切的联系因此,了解药物对CYP450酶可能存在的诱导或抑制作用是药物研究过程中不可缺少的研究内容本文综述了近年来研究CYP450酶诱导/抑制水平和机制的试验方法以及在药物尤其是在天然产物研究中的应用细胞色素P450酶;诱导/抑制;研究方法;药物开发;天然产物:R969.1:A:10092501(2011)05057907,,6:P450;(NAT)(MT)CYP450,90%CYP450,()CYP450,CYP450(/);,CYP450,,,,CYP450[1]CYP450/1CYP450/CYP450/,,,CYP450,NCYP2E1[2],,579CYP,,CYP450,,CYP450/,CYP450Westernblot,,ACYP1A2CYP2E1,[3],,,mRNA/0.1mmol/LCYP1A22D6,,[4],,,[5]P450,/CYP450,/,LiuY[6]CYP450Rb1Rb2RcRdReRg1Rg2SRg3Rh1Rh2F1CKPpdPpt,CKPpdPptCYP2C9;PpdPptCYP3A4CYP450,,,CYP450CYP450,/,CYP450[7],,80Cocktail,,,,3CYP1A22D63A4,3CYP2D6,AUC[8][9]CYP4505(CYP1A2)(CYP2D6)(CYP2C9)(CYP2C19)(CYP3A4)5(CYP1A2)(CYP2D6)(CYP2C9)(CYP2C19)(CYP3A4),LC/MS5(451),55,,IC50,,CYP4502CYP450/CYP450/,,/,CYP450/CYP450:(1)DNAAh,,CYPDNA(2)mRNA/(3)CYP,580ChinJClinPharmacolTher2011May;16(5)(4),CYP,,,CYPCYP1A2mRNA,CYP1A2,[10]X(PXR)mRNACYP3A4CYP3A4,2PXRmRNA,DMAIIIRXR[11],CYP450,CYP4505:(AhR),CYP1A1CYP1A2;,CYP2E1;(PPAR),CYP4A;CYP2B3A1A2;PXRCYP3A(budesonide),PXRCYP3A4[12]PXRHepG2CYP3A4;IIAPXR,CYP3A4[13]3,AhRCYP1A1,[14]PXR,CYP3A4,(DEHP)CYP3A4PXR,CYP3A4[15]CYP450[16]:(1)(2)CYP450,CYP450,CYP450,,(3),(4)CYP450MICYP(5)CYP[17]HPLC(),6CYP450,50mol/LCYP1A2CYP2E1CYP2C19CYP2D6Yuka[18]60CYP3A42C9,,LineweaverBurk,CYP3A42C9,CYP2C9KiCYP2C9B33.1(transienttransfectionassays),DNA/RNA,,,24~72h(),:(1);(2);(3);(4);(5)[19],HepG2(GBE)(BB)A(GA)B(GB)PXR5812011May;16(5)CYP3A4[20]HepG2,PXRCYP3A4,WangK[21]HepG2Caco2,FuGENE6tk(3A4)3Luc,1248h,9cis,9cisRARARXR/VDRChenSY[22]RXRCARtk(3A4)3LucHepG2,48hDMSOTCPOBOP,TCPOBOP5(9cis,9cisRA,13cisRA,RA,RP)RXR/CARRXR/CAR,9cisRARARXR/CAR3.2(electrophoreticmobilityshiftassay,EMSA)DNA,DNAIstrateM[23]PXRCYP3A4,CYP3A45'PXRER6TR1VDRTR1PXRCARFXRCOUPTFLXR,DR3VDRCOUP-TFCOUP-TFCARTR1PXRTR1LXRFXR,PXRKouichi[24](halfsite)hCARhRXR,hCAR/hRXRER8Toriyabe,PXR,CYP3A4,,hPXR/hRXReNR3A4(essentialdistalnuclearreceptorbindingelementforCYP3A4induction),hPXR/hRXRCYP3A43(dNR1prER6eNR3A4),CYP3A4,eNR3A4,[25]PavekP[26]eNR3A4LS174TPXRCYP3A43.3(reportergeneassay),(DNA)(),,,:(LUC)(gal)(CAT)(GUS)(SEAP),,,,,Kouichi[24]HepG2,hCAR511554;AhRCARCYP1A1CYP1A2miR148aPXRmRNAmiR148a,PXR;PXRmiR148aPXRCYP3A4mRNA,miR148ahPXRCYP3A4[27]3.4(chromatinimmunopre582ChinJClinPharmacolTher2011May;16(5)cipitationassay,ChIP)DNA,DNA,ChenS[22]ChIP9cis9cisRARXR/CARCYP3A4YoshinariK[24]EMSA,ChIPHepG2pcDNAhCAR,hCARCYP1A1CYP1A2hCARhRXRCYP1A1ER8,AhRRothA[28]ChIPCyp2b10,1(SREBP1)SREBP1DNA,PXRCAR(1),CYPs,,DNArealtimePCRWesternblot,LiuF[29]FER6HNF4,PXRCYP3A4,PXRFER6,HNF4,,HNF4CYP3A4CervenyL[30](VPA)PXRCARCYP3A4VPACAR/RXRCYP3A4DR3,PXRCYP3A4;,VPACYP3A4mRNA,CYP3A4FangH[31]23,CYP3A4,12SULT2A1mRNA,(11),SULT2A15'HepG2,SULT2A1HNF4;PXRSULT2A1HNF4,HNF4,SULT2A14CYP450,CYP450/,,,/,;/,,,,mRNA1~3h,/,72hEMSAChIPDNA,EMSA,;ChIP,,,,,CYP4505832011May;16(5),,,CYP450,,CYP450/[1],.P450:[J].,2008,35(1):62-64.[2],.P450[J].,2008,21(3):132-134,145.[3],,,.-AP450[J].,2009,40(6):938-942.[4],,,.P450[J].,2008,11(2):131-134.[5],,,.[J].,2007,24(1):32-36.[6]LiuY,ZhangJ,LiW,etal.Ginsenosidemetabolites,ratherthannaturallyoccurringginsenosides,leadtoinhibitionofhumancytochromeP450enzymes[J].ToxicolSci,2006,91(2):356-364.[7],,,.P450[J].,2010,15(12):1330-1333.[8],,.CYP1A2CYP2D6CYP3A4[J].,2010,15(7):775-780.[9],,,.P4505[J].,2007,42(6):589-594.[10],,,.CYP1A1[J].,2006,(10):1241-1242.[11]MedinaDazIM,EstradaMu?izE,ReyesHernndezOD,etal.Arseniteanditsmetabolites,MMAIIIandDMAIII,modifyCYP3A4,PXRandRXRalphaexpressioninthesmallintestineofCYP3A4transgenicmice[J].ToxicolApplPharmacol,2009,239(2):162-168.[12]ZimmermannC,WaterschootR,HarmsenS,etal.PXRmediatedinductionofhumanCYP3A4andmouseCyp3a11bytheglucocorticoidbudesonide[J].EurJPharmSci,2009,36(4/5):565-571.[13]YuCN,YeSS,SunHY,etal.PXRmediatedtranscriptionalactivationofCYP3A4bycryptotanshinoneandtanshinoneIIA[J].ChemBiolInteract,2009,177(1):58-64.[14]WeemsJ,YostG.3MethylindoleMetabolitesInduceLungCYP1A1andCYP2F1EnzymesbyAhRandNonAhRMechanisms,Respectively[J].ChemResToxicol,2010,23(3):696-704.[15]CooperB,ChoT,ThompsonP,etal.PhthalateinductionofCYP3A4isdependentonglucocorticoidregulationofPXRexpression[J].ToxicolSci,2008,103(2):268-277.[16],.P450[J].,2006,8(3):333-335.[17],,,.P450[J].,2005,3(4):243-245.[18]KimuraY,ItoH,OhnishiR,etal.InhibitoryeffectsofpolyphenolsonhumancytochromeP4503A4and2C9activity[J].FoodChemToxicol,2010,48(1):429-435.[19],,,.XCYP3A4[J].,2009,20(3):234-237.[20],,,.PXRP4503A[J].,2008,24(4):504-509.[21]WangK,ChenS,XieW,etal.RetinoidsinducecytochromeP4503A4throughRXR/VDRmediatedpathway[J].BiochemPharmacol,2008,75(11):2204-2213.[22]ChenS,WangK,WanY.RetinoidsactivateRXR/CARmediatedpathwayandinduceCYP3A[J].BiochemPharmacol,2010,79(2):270-276.[23]IstrateM