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药物代谢及其动力学在新药研发中的应用胡卓汉博士瑞德肝脏疾病研究(上海)有限公司复旦大学药学院2004年12月30日中国.北京EfficacyHitsOptimizedLeadGoornogodecisionCompoundforDevelopment(CD)NEWDRUGINDNDAR&D临床实验临床前实验研究和发现药物研发的三大任务•药效Efficacy/Pharmacodynamics•安全Safety/Toxicology•药物代谢动力学DrugMetabolism/Pharmcokinetics药物代谢动力学的任务0.010.111010010000.511.522.533.54.（最大无毒性浓度）(最小有效浓度）(最小药效时间）血浆浓度时间药效毒理药代最佳血浆浓度EfficacyHitsOptimizedLeadGoornogodecisionCompoundforDevelopment(CD)NEWDRUGINDNDAR&D临床实验临床前实验研究和发现研究和发现阶段•能否被吸收？permeability•是否被代谢？metabolicstability•代谢产物？metaboliteidentification•代谢途径？pathwayidentification•对其它药物的影响？drug-druginteractionEfficacyHitsOptimizedLeadGoornogodecisionCompoundforDevelopment(CD)NEWDRUGINDNDAR&D临床实验临床前实验研究和发现临床前阶段•生物利用度bioavailability•血浆浓度的线性和非线性doseescalation&proportionality•多次给药和体内积蓄multipledoses&accumulation•吸收和排泄模式massbalance•体内分布distribution•从动物代谢推算人体代谢extrapolationEfficacyHitsOptimizedLeadGoornogodecisionCompoundforDevelopment(CD)NEWDRUGINDNDAR&D临床实验临床前实验研究和发现临床阶段•长期毒性实验的动物选择metabolismprofilinginanimalsandhumansEfficacyHitsOptimizedLeadGoornogodecisionCompoundforDevelopment(CD)NEWDRUGINDNDAR&D临床实验临床前实验研究和发现临床实验准则GoodClinicalPractice(GCP)非临床实验准则GoodLaboratoryPractice(GLP)二五原则•5毫克•5天临床前实验药物代谢动力学的生物模型体外和离体模型(invitro/insitumodels)•吸收模型absorption/permeability•代谢模型metabolism•体外推测和体内(invitro/invivocorrelation)动物模型(invivoanimalmodels)动物推测人(speciesextrapolation)排出太快/药效时间太短口服吸收差/血浆浓度太低分布排泻代谢问题吸收问题蛋白质相互作用分布体积肾脏排泄肝脏代谢溶解度肠道吸收膜通透性肠道消化早期研发阶段后期研发阶段SituationAnalysisinvitro体外metabolisminsitu离体permeabilityinvivo体内bioavailabilityPlasmaconcentrationsofBCH-3840anditsmetabolite(BCH-6440)inmicedosed50mg/kgorallyPoororalbioavailability06012018002004006008001000ParentMetaboliteTmax(min)6.146.53Cmax(g/ml)0.0830.799AUC(g/ml/min)4.82321.810Bioavailability%2.52311.434R20.92920.9964BCH3840MetaboliteTime(min)Concentration[ng/ml]药物吸收模型计算机脂溶度脂层转移细胞层转移十二指肠灌流14absorption/distributionmodel脂层转移模型水相Aqueousphase水相Aqueousphase有机相OrganicphasepH=6.5pH=7.4PermeabilityEvaluation–invitro15invitroabsorption/distributionmodelInVitro/InVivoCorrelationPooledDatafromFourBiostudies020406080100020406080100formulationfindingstudyBEstudy1BEstudy2Y=4.2+1.00X,R2=.987specificationstudy%Distributed%AbsorpedCaco-2TransportPathways人大肠癌细胞模型TransportPathways药物吸收机制被动细胞间主动P糖蛋白ProbesforTransportPathways肠道吸收标准对照药物•Transcellular（被动吸收）Propranolol,Testosterone•Paracellular（细胞间渗透）Mannitol,Inulin•Carriermediated（主动吸收）Glucose•P-Glycoproteinmediated（P－糖蛋白调节）底物Vinblastine抑制物VerapamilGlucose（蔗糖）vsInulin（木香素）主动吸收vs细胞间渗透050100150200250020406080100Time(min)Flux050100150200250300350400450Papp(nm/sec)day18/23/991999-8-301999-9-131999-9-211999-9-281999-10-51999-11-11999-11-81999-12-61999-12-131999-12-202000-1-102000-1-172000-1-24TestDayMannitolPropranololPropranololvsMannitol被动吸收vs细胞间渗透020406080100120Papp(nm/sec)day18/23/991999-8-301999-9-131999-9-211999-9-281999-10-51999-11-11999-11-81999-12-61999-12-131999-12-202000-1-102000-1-172000-1-24TestDayVinblastineVinblastine/verapamil由P－蛋白所调节的药物吸收－使用P－糖蛋白抑制剂VerapamilOralAbsorption3-dayDrugsinHumansCaco-2(%)Kpcaffeine100227ibuprofen100201desipramine95261acetaminophen95218propranolol90265hydrazine90155Ketoconazole76120terbutaline7356atenolol5030acetbutalol4029nadolol3522losartan3342mannitol1640inulin512Chong,Dando&Morrison;Pharm.Res.1997FalsePositive假阳性＝低FalseNegative假阴性＝高Caco-2TransportPathways人大肠癌细胞吸收模型insituratintestinalperfusion(singlepass)离体大鼠十二指肠灌流模型（单循环）METHODAnimal:MaleSprague-Dawleyrats(250-350g),fastedovernight.Ratisanesthetizedbyurethane1.5g/kg,im.beforeperfusionstarts.Perfusate:Phosphatebuffer,pH=6.510mMglucosePhenolred(negativecontrol)Acetaminophen(positivecontrol)Finalconcentrationsoftestarticle=0.05-0.30mg/mLPerfusionProcedures:•ratisputonaheatingpadtomaintainbodytemperature•jejunumisexposedviaamiddlelineincision•sutures:1stismadeat5cmdistaltotheligamentofTreitz2ndismadeatabout20cmdistalto1stone•theinletofcannula-asyringeinfusionpump•theoutletofcannula-afractioncollector•theperfusionsegmentisprecleanedbypassing10mlofblankperfusatebuffer•perfusiontimeandrate=0.1ml/minfor120min•outletperfusionsamplesarecollectedevery10min•plasmasamplesarecollectedat30,60,90and120minafterperfusionCalculations:Permeability(Peff,cm/min)=(Q/2RLp)x(1-C’out/C’in)C’out/C’in=(Cout/Cin)x[phenolred]in/[phenolred]outinsituratintestinalperfusion(singlepass)Insituratintestinalpermeability(singlepass)0.0000.0010.0020.0030.0040.005020406080100Permeability(cm/min)HumanOralBioavailability(%)Predictionwithin90%interval=19/31(61.3%)In-housevalidation假阳性假阴性PlasmaconcentrationsofBCH-3840anditsmetabolite(BCH-6440)inmicedosed50mg/kgorallyPoororalbioavailability06012018002004006008001000ParentMetaboliteTmax(min)6.146.53Cmax(g/ml)0.0830.799AUC(g/ml/min)4.82321.810Bioavailability%2.52311.434R20.92920.9964BCH3840MetaboliteTime(min)Concentration[ng/ml]排出太快/药效时间太短口服吸收差/血浆浓度太低分布排泻代谢问题吸收问题蛋白质相互作用分布体积肾脏排泄肝脏代谢溶解度肠道吸收膜通透性肠道消化早期研发阶段后期研发阶段SituationAnalysisinvitro体外metabolisminsitu离体permeabilityinvivo体内bioavailabilityInSituRatIntestinalPermeability:Good50607080901001101201301401500255075100PhenolRedAcetaminophenBCH-3840