脂质体制备方法的里程碑-挤出法-蒋宫平

[e-mail]*[e-mail]。1006-2858201308-0646-07-*110016。32、、、、、。、Tm。、、、、。Ｒ94A1Bangham119652－3。1974Ｒahman45、。～100nmEPＲ。6、7、French8、。1979Olson7Hope198691989Cullis101990Cullis111992Lentz12PEG。、、、13、。2。3NorthernLipidsAvantiPolarLpidAvestin0.25mL、。、。MacDonald13。1ABCDOE。O。30820138JournalofShenyangPharmaceuticalUniversityVol.30No.8Aug．2013p.646。1.27～2.54cm1.27cm1.91cm。1mL。MacDonald0.25mL0.5mL2。。NorthernLipidsTheLI-PEXTMEXTＲUDEＲS15～20mL800psi1±20%。、。Fig.1Thestructureofminiextrusiondevice14。phasetransitiontem-peratureTm。TmC-C、2。Fig.2Thereleaseofdrugspermeatinglipidmembranesatthetemperaturehighorbelowthephasetransitiontemper-atureTm2Tm7468-。Tm。TmTm。80℃。55.1Olson7phosphatidylserinePS-phosphatidylcholinePC-cholesterolCH1∶4∶5multila-mellarvesicleMLV1.0、0.8、0.6、0.40.2μm1.80μm0.27μm7。10～12mmol·L－150lb·inch－20.2μm60mmol·L－1。140.7077。3S=2πr2S=πＲ2S=S2πr2=πＲ2r/Ｒ=0.707。、。Fig.3Theprincipleofnegativestainelectronmicrosco-pytocalculatethesizeofliposome3hydrogenatedsoybeanphosphatidylcholineHSPC-CH--polyethyleneglycol-distearoylphosphatidylethano-laminePEG－DSPE3.0∶0.5∶0.320g·L－10.45、0.220.10μm143.5nm106.5nmCoeff.ofVar'n0.4850.314Chisq1.280.72。。5.215averagenumberofbilayersn。n31P-NMＲ、freeze-fractureelectronmicroscopyFFEMn15。5.2.131P-NMＲSpies150.5mL2H2OBrukerWH9018W25℃。1.5mL60μL100mmol·L－1MnCl2。1631P。Mg2+percentagerelativelossofsignalＲLOSn1。n=100/2×ＲLOS1。1n。1。n1。5.2.2BalzersLichtenstein。30%HClOreplicasEM300n。particledensitypdπ×Dsurf2Dsurf=averagesurfacediame-ter。phospholipidconcentrationPLC84630surfaceareapermolePLinthebilayerBSAn2。n=BSA×PLCpd×2π×Dsurf22Pidgeon17encapsu-latedvolumeE、lipidvolumepermolePLLV/volume/surfaceareavsto-talsurfaceareaTSA。TSABSAn3。vs=totalvesiclesvolumetotalsurfacearca=E+LVTSA3TSA=BSA2×n3n=vs×BSA2E+LV3Olson1、23n。n、、、。n15。nMLVnn。5.3。1993Cullis18“”。largeunilamellarves-icleLUVslysis。2.35mmol·L－1NaCl100nmcarboxyfluoresceinCF20mmol·L－1HEPESpH7.4100nmeggphosphatidylcholineEPC-CH55∶45。Cullisap-pliedosmoticdifferentialAODresidualosmoticdifferentialＲOD。4ＲODＲODAOD。AOD1780mOsm·kg－1。AODＲOD650mOsm·kg－1。12nmdefect18ＲOD。Fig.4Influenceoftheosmoticdifferentialonvesiclelysis4Sun19ＲOD。τ=ΔPr/2τΔPＲODr。。τＲODr－10518。40dyn·cm－1Needham20。5.4nthewaterpermeabilitykwthepermeabilityofaminoacidkA。Morii21PCCH20mmol·L－14mmol·L－1pH7.4Pipes-TrisAMLVreverse9468-phaseevaporationvesiclesＲEVKwKA。Fig.5Ｒelationshipbetweenvesicleradiusandosmotictolerance5ＲOD5.4.1100mmol·L－11∶122－23。MLV0.8μmＲEV0.38μmKw210.2μmKwKw24CHＲEVKwCH25－27。5.4.2B100mmol·L－1-A1∶122－23。0.08μm5glycineGly、prolinePro、valineVal、leucineLeuphenylalaninePheKAKA-Phe4.2KA-Gly200。Morii21KA15KA。5.5encapsulatedvolumeEVlipidL·μmol－1L·kg－1%28－29。E。EE。E、。Jousma15－30MLVsCF31SephadexG50CF/491nmCFFiske32E1。MLVs600nm5E2.5L·mol－13.2L·mol－13a.Eb.LUVLUVMLVEc.E。MLVs600nm1200nm5E2.5L·mol－12.0L·mol－12a.Eb.15E。Table1Theencapsulatedvolumeobtainedfromlipo-somespreparedbyextrusionthroughdifferentmem-braneporesizeandtimes1No.ExtrusionmethodEV/L·mol－11Noextrusion2.72600nmextrusion1cycle2.53600nmextrusion5cycles3.24600nmextrusion1cycleand2.0200nmextrusion5cycles630、、。05630。1BANGHAMASTANDISHMWATKINSJ．Diffu-sionofunivalentionsacrossthelamellaeofswollenphospholipidsJ．JournalofMolecularBiology1965131238－252．2LASICDD．NovelapplicationsofliposomesJ．TrendsinBiotechnology1998167307－321．3LASICDD．LiposomesfromphysicstoapplicationsM．NewYorkElsevierAmsterdametc1993．4ＲAHMANYECEＲNYEATOLLAKSENSLetal．Liposome-encapsulatedactinomycinDpotentialincancerchemotherapyJ．ProceedingsoftheSocietyforExperimentalBidogyandMdicine197414611173－1176．5AMSELEMSGABIZONABAＲENHOLZY．Opti-mizationandupscalingofdoxorubicin-containinglipo-somesforclinicaluseJ．JournalofPharmaceuticalSciences199079121045－1052．6SENIOＲJ．FateandbehaviorofliposomesinvivoareviewofcontrollingfactorsJ．CriticalＲeviewsinTherapeuticDrugCarrierSystems198732123．7OLSONFHUNTCSZOKAFetal．PreparationofliposomesofdefinedsizedistributionbyextrusionthroughpolycarbonatemembranesJ．BiochimicaetBiophysicaActa197955719．8BＲANDLMBACHMANNDDＲECHSLEＲMetal．Liposomepreparationusinghigh-pressurehomoge-nizersJ．LiposomeTechnology1993149－65．9MAYEＲLDBALLYMBHOPEMJetal．Tech-niquesforencapsulatingbioactiveagentsintolipo-somesJ．ChemistryandPhysicsofLipids1986402/4333－345．10NAYAＲＲHOPEMJCULLISPＲ．Generationoflargeunilamellarvesiclesfromlong-chainsaturatedphosphatidylcholinesbyextrusiontechniqueJ．Bio-chimicaetBiophysicaActaBBA-Biomembranes19899862200－206．11LOUGHＲEYHWONGKCHOILetal．Protein-li-posomeconjugateswithdefinedsizedistributionsJ．BiochimicaetBiophysicaActaBBA-Biomem-Branes19901028173－81．12LENTZBＲMCINTYＲEGFPAＲKSDJetal．Bi-layercurvatureandcertainamphipathspromotepolyethyleneglycol-inducedfusionofdipalmitoylphos-phatidylcholineunilamellarvesiclesJ．Biochemis-try199231102643－2653．13MACDONALDＲCMACDONALDＲIMENCOBPMetal．Small-volumeextrusionapparatusforprep-arationoflargeunilamellarvesiclesJ．BiochimicaetBiophysicaActaBBA-Biomembranes199110612297－303．14PAPAHADJOPOULOSDMILLEＲN．Phospholipidmodelmembranes．I．Structuralcharacteristicsofhy-dratedliquidcrystalsJ