白蛋白纳米球药物载体的制备及表征

42Vol.4No.220044TheChineseJournalofProcessEngineeringApr.2004(100080)−60∼100nm....16:1.Higuchi.R944.2A1009−606X(2004)02−0155−051.[1]..200nm[2].[3].(albuminnanosphere).5µm,[1,3][4−7]1µm200nm.Mller[3−5]200nm.−60∼100nm.22.1(BSA)(Amresco)(Amresco,1:250)25%(Sigma)(10mg).Arlacel83(Sigma)HCO−40(40Nikkol)(SigmaTypeII−S),Span85(2003−05−282003−07−18(20125616)(1979−)Tel:010−82627072.1564)Span80()..JY92−2D()H−600()Du7500(Beckman)THZ−C()DZF−6020().2.21:50.234mol/L.1ml(20%0.25%)50ml(2%)600r/min1h,10min300W.2h1h.2324h.2.32.3.1.n(n200)di(nm)d(nm)CV(coefficientofvariation)1221()1CVniidddn=−=∑.2.3.250mg5000r/min5min−=×100%.2.3.3C0.2,0.5,1,2,4,6,8,10,12,14,16,18,20mg/L365nmAA=0.04222C+0.00176,R=0.99977.2mg4mg10mlpH7.437oC.10ml..2.3.430mg5ml25ml37oC2ml2ml.33.1Durbin[9]Landfester[10−11].2157...5W/O1.5ml20%BSA4ml90s(80W),1.Arlacel83HCO−40.HCO−40Arlacel832%.1Table1Timeforphaseseparationinemulsionspreparedwithvariousemulsifiers(min)EmulsifierEmulsifiercontent0.5%Emulsifiercontent5%HCO-4045420Span8045240Lecithin530Span85530Arlacel83454203.22BSANH2+CHO(CH2)3CHO→BSANCH(CH2)3CH==NBSA,.2.16:1.2Table2InfluenceoftheamountofglutaraldehydeonpropertiesofalbuminnanospheresRatioofaldehydegrouptoaminogroup(moleratio)Diameterofnanosphere(nm)CV(%)Yield(%)Drugcontent(%)Drugloadingefficiency(%)Swellingdegree(%)1:199.3434.6854.060.7928.45268.784:195.2731.2565.120.8034.53207.238:186.2530.0670.531.1554.24119.7312:172.8529.6481.241.6690.15107.8616:167.6336.3797.611.2983.67112.10..2(1).(2)..100nm1TEMFig.1TEMphotographofalbuminnanospheres(−CHO:−NH2=16:1)16:1TEM1.1584[3−5](300W)(10min),.100%[1]90.38%[8]75%90.15%.2.5h...[1](560nm)5h19%55h18.6%∼24.5%.020406080100010203040506070Releasedfraction(%)Time(h)CHO:NH21:14:18:112:116:12Fig.2Invitroreleaseofmitomycinfromthealbuminnanospheresmadewithdifferentamountsofglutaraldehyde(16:1)Higuchi3Qt.Higuchi.3Table3ThefittingofthereleasecurveofalbuminnanospheresFittedequationModel0∼5h5∼92hZero-orderkineticsQ=0.0359t+0.0257,R=0.9413Q=1.44×10−3t+0.236,R=0.8970First-orderkinetics1−Q=0.975e−0.04041t,R=0.94691−Q=0.764e−0.00202t,R=0.9081HiguchiequationQ=0.08427t1/2+0.00177,R=0.9685Q=0.01857t1/2+0.1859,R=0.9540Higuchi4.RHiguchi.Higuchi.4HiguchiTable4TheHiguchiequationfittingofthereleasedfractionofdrugfromalbuminnanospherespreparedwithdifferentamountsofglutaraldehydeFittedequation−CHO:−NH20∼5h5∼92h1:1Q=0.1075t1/2+0.0133,R=0.9863Q=0.0584t1/2+0.1257,R=0.99044:1Q=0.0789t1/2+0.0258,R=0.9136Q=0.0501t1/2+0.0402,R=0.98308:1Q=0.0834t1/2+0.0159,R=0.9692Q=0.0375t1/2+0.111,R=0.994112:1Q=0.0801t1/2+0.0092,R=0.9671Q=0.0285t1/2+0.1313,R=0.99304−(300W)(10min)2159(Arlacel83)60∼100nm...Higuchi..[1],.[J].,1999,30(5):353−356.[2].[M].2002.63.[3]MllerBG,LeuenbergerH,KisselT.AlbuminNanospheresasCarriersforPassiveDrugTargeting:AnOptimizedManufacturingTechnique[J].Pharm.Res.,1996,13(1):32−37.[4]LinW,GarnettMC,SchachtE,etal.PreparationandinvitroCharacterizationofHSA−mPEGNanoparticles[J].Int.J.Pharm.,1999,189:161−170.[5]LinW,GarnettMC,DavisSS,etal.PreparationandCharacterizationofRoseBengal-loadedSurface-modifiedAlbuminNanoparticles[J].J.Control.Rel.,2001,71:117−126.[6]CoombesAGA,BreezeV,LinW,etal.LacticAcid-stabilisedAlbuminforMicrosphereFormulationandBiomedicalCoatings[J].Biomaterials,2001,22:1−8.[7]ChenGO,LinW,CoombesAGA,etal.PreparationofHumanSerumAlbuminMicrospheresbyaNovelAcetone-heatDenaturationMethod[J].J.Microencapsul.,1994,11(4):395−407.[8]GuptaPK,HungCT,LamFC.FactorialDesignBasedOptimizationoftheFormulationofAlbuminMicrospheresContainingAdriamycin[J].J.Microencapsul.,1989,6(2):147−160.[9]DurbinDP,El-AasserMS,PoehleinGW,etal.InfluenceofMonomerPreemulsificationonFormationofParticlesfromMonomerDropinEmulsionPolymerization[J].J.Appl.Polymer.Sci.,1979,24:703−707.[10]LandfesterK,WillertM,AntoniettiM.PreparationofPolymerParticlesinNonaqueousDirectandInverseMiniemulsions[J].Macromolecules,2000,33:2370−2376.[11]WillertM,LandfesterK.AmphiphilicCopolymersfromMiniemulsifiedSystems[J].Macromol.Chem.Phys.,2002,203:825−836.[12]FujimotoS,MiyazakiM,EndohF,etal.MitomycinCarryingMicrospheresasaNovelMethodofDrugDelivery[J].CancerDrugDeliv.,1985,2(3):173−181.PreparationandCharacterizationofAlbuminNanospheresasDrugCarrierWANGKai,MAGuang-hui(StateKeyLab.Biochem.Eng.,Inst.ProcessEng.,ChineseAcademyofSciences,Beijing100080,China)Abstract:Mitomycin-loadedalbuminnanospheressizedbetween60and100nmwerepreparedbytheultrasonicemulsification−crosslinkingmethod.Afterthealbuminaqueousphasecontainingmitomycinwasemulsifiedintotheoilphasebytheultrasonifiertoobtainthenanodroplets,glutaraldehydewasaddedtocrosslinkthenanodroplets.Theoptimalemulsifierwaschosenbycomparingthestabilityofemulsionpreparedwithvariousemulsifiers.ItwasfoundthatthemoststableW/OemulsionwasobtainedwhenArlacel83wasusedastheemulsifier.Theinfluenceoftheamountofglutataldehydeonthepropertiesofthealbuminnanosphereswasinvestigated.Itwasfoundthattheyieldanddrugcontentofnanospheresandloadingefficiencyofthedrugincreased,whilethediameterandswellingdegreeofnanospheres,andtherateofdrugreleaseinvit