药用高分子材料siRNA药物专家讲座

姚老师作业：聚乳酸本体聚合工艺，对应机理和反应方程第1页第六章高分子纳米药品-siRNA药品病变组织基因活性正常表示过分表示低表示小分子抑制剂?蛋白疗法?抑制剂:RNA干扰疗法,反义RNA,…下调上调增强表示:

质粒DNAViralVectorsEmulateDrug?第2页

抗体:Bindprotein,butonlyextracellularepitopesLowthroughput,highcost反义RNA/核酶:Stopgeneexpression(production),mostproteinsChemicalstabilizationessentialBarriers:sequenceselection,potency,toxicities,…

RNA干扰:Stopgeneexpression(production),mostproteinsGoodactivitywithoutchemicalstabilizationRapidlyreplacingantisense:reallybetter?特异性基因下调伎俩第3页RNA干扰（RNAinterference,RNAi）技术功效：特异性地缄默内源或外源性靶基因影响：年《Science》杂志十大科学成就之二 年《Science》杂志十大科学成就之首医学领域应用：基因功效研究；基因敲除；基因治

疗；基因表示调控等;

抗病毒以及糖尿病、肥胖症和癌症、 黄斑变性眼科疾病治疗等。第4页RNAi作用机制Nature，，430：161第5页Log[Oligo](M)NormalizedGeneExpressionLevelAntisensesiRNAControlsFromH.Winkler,IBCConference“RNAInterference”,SanDiego,CA,Feb第6页siRNAdsRNAsiRNARNAseshRNAInVitroInVivoInVivomiRNAAdaptedFromDykxhoornetal.,NatureReviews4,457,含有RNAi机制药品第7页TargetIDsiRNATargetValidationGrowingClinicalLeadDiscovery&OptimizationPreclinical药品递送是关键挑战siRNA药品?siRNA药品:革命性突破第8页

Company

siRNA

Disease

Routeofadministration

Stage

ALN-TTR01ATTRintravenousALN-RSVRSVinfectionIntranasalALN-VSPLiverCancerIntravenousQPI-1007NAIONIntravenousI5NPARFIntravenousPF-4523655AMDIntravitrealBevasiranibAMDIntravitrealDMEIntravitrealTKM-080301CancerIntravenousPRO-040201HypercholesterolemiaIntravenous

ATU027AdvancedSolidTumorsIntravenoussiG12DLODERPancreaticCancer

intratumoralTD101PCIntradermalsir027AMDIntravitreal

RTP801i-14AMDIntravitreal

CALAA-01SolidtumoursIntravenousPotentialofsiRNA-BasedTherapyPhase1Phase2Phase3RNA干扰疗法潜力siRNA药品快速进入临床研究第9页难以被细胞摄取：大分子，阴离子脱靶效应：非特异性缄默，引发免疫刺激等难以富集在肿瘤部位：体内不稳定，半衰期短

1

2

3PotentialofsiRNA-BasedTherapyRNA干扰疗法潜力siRNA药品临床引用关键第10页Nature,468,p487,Nov.25,PotentialofsiRNA-BasedTherapyRNA干扰疗法潜力siRNA给药技术成为关键挑战第11页表示siRNA质粒DNA传递病毒（NatureBiotechnology,,20:1006-10）非病毒类载体化学合成siRNA传递 非病毒类载体：脂质体和高分子 物理伎俩:electroporation,highpressure/highvolumeinjectionPotentialofsiRNA-BasedTherapyRNA干扰疗法潜力siRNA传递：体内RNAi技术应用关键第12页脂质类Figure2.ChemicalmodificationsofsiRNAs.Shownarestructuresofsugar,backboneandbasemodificationsandofthecholesterolconjugate.NatueChemicalBiology,2(),711-719.Naturereviewsdrugdiscovery,

6(),443-453高分子纳米载体抗体类化学修饰siRNAsiRNA缀合物PotentialofsiRNA-BasedTherapyRNA干扰疗法潜力siRNA非病毒载体种类第13页内涵体逃避靶向结合吞噬核转运DNA释放到细胞质形成RISC内涵体逃避靶向结合吞噬siRNA释放到细胞质siRNA传递DNA传递组织转运DAN和siRNA传递比较蛋白质表示转录mRNA第14页化学修饰siRNA第15页Figure2.ChemicalmodificationsofsiRNAs.Shownarestructuresofsugar,backboneandbasemodificationsandofthecholesterolconjugate.NatueChemicalBiology,2(),711-719.第16页Naturereviewsdrugdiscovery,

6(),443-453Nature，，432：173-178

化学修饰siRNA第17页Challenge1:稳定Challenge2:长循环Challenge3:靶向Challenge4:渗透Challenge5:细胞摄取Challenge6:制剂化基于纳米载体siRNA给药系统第18页EngineeringNanoparticlesforsiRNADeliveryExtendedCirculationTissueTargetingPenetrationCellinternalization第19页脂质体类siRNA纳米载体第20页脂质体类siRNA纳米载体第21页StableNucleicAcidLipidParticle(SNALP)第22页StableNucleicAcidLipidParticle(SNALP)Meanparticlesize:140±12nmMeanencapsulationefficiency:93%±3%Nucleicacid/lipidratio:42±4

g/

mol第23页siRNA-SNALPwithalongerhalf-lifeinplasmaandliverreducedserumHBVDNAtitersinmousemodelfollowingintravenousinjectionNatureBiotechnology

23,1002-1007()第24页ApoB-siRNA-SNALPresultedinsilencingofApoBmRNAexpressioninliverandsignificantreductionsinApoBprotein,serumcholesterolandlow-densitylipoproteinlevelsNature441,111-114(4May)第25页高分子类siRNA给药系统第26页StericCoat(Inert)ActiveCoreExposedTargetingLigandsCleavableLink(Shedding)LigandsStericpolymerCarrierCleavablelinkage+++++++siRNAPayload水溶性阳离子高分子纳米载体:第27页NUCLEICACIDSRESEARCH32(19):Art.No.e149第28页PNAS,104,5715-5721靶向修饰水溶性阳离子高分子纳米载体:第29页CancerResearch,65,8984-8992第30页LigandPEGDegradablelinkagesiRNAJACS,127,1624-1625靶向修饰水溶性高分子-siRNA键合物:第31页第32页JACS,126(42),13612-13613第33页LoadingsiRNApostnanoparticleformationPEG:prolongedbloodcirculationBiocompatibleDegradable,potentiallyintracellularsiRNAreleaseCo-deliveryofchemotherapeuticdrugforsynergisticeffect++++++++++++++++++++++++++++++siRNA亲水嵌段疏水嵌段带正电荷嵌段阳离子胶束/siRNA复合纳米颗粒:第34页abc30minFAM-siRNA12h24hRho-PTX30min12h24hRho-PTXMicelleplexFAM-siRNAFAM-siRNAPBSabcdeabcdeabcdeRho-PTXMicelleplexNCsiRNA+MicelleplexFAM-siRNAbcdeaSunTM,etal.,ACSNano,,5,1483-1494第35页Dayspost-xenografting121620242832364044TumorVolume(mm3)100200300400500600PaclitaxelMicelleplexNCsiRNAPaclitaxelMicelleplexsiPlk1PaclitaxelMicelleplexNCsiRNA+MicelleplexsiPlk1MicelleplexsiPlk1PBSBlankMicellesMicelleplexNCsiRNASunTM,etal.,ACSNano,,5,1483-1494β-actin020406080100120Plk1mRNA(%ofControl)MicelleplexNCsiRNAPBSBlankmicellesMicelleplexsiPlk1PaxlitaxelMicelleplexPaxlitaxelMicelleplexsiPlk1PaxlitaxelMicelleplexNCsiRNA+MicelleplexsiPlk1Plk1第36页KimA.Woodrow,W.MarkSaltzmanetal.,NatureMaterials,,8,526–533ChristopherJ.Cheng,

W.MarkSaltzman,