半乳糖化阿昔洛韦三节前药纳米给药系统的研究

广东药学院硕士学位论文半乳糖化阿昔洛韦三节前药纳米给药系统的研究姓名：洪华云申请学位级别：硕士专业：药剂学指导教师：朱亮20090501IANPGANPN-NHSNN'-DCCSACVSACVHSBSA-ACVpHHepG-2SACVNHSSACVDCCSACVBSA-ACVBSApHHPLCBSA-ACVpHBSA-ACVBSA-ACVpHBSA-ACV-ANPMTTGANPHepG-2FTIRSACVNHSIISACV79.1%BSA-ACV24.1%HPLCBSA-ACVpHpH2.0t1/258.73hpH9.7t1/25.41hTEMLPA187nm4.22%48h56.19%HepG-248h96.5%0.954mg/mL8.853mg/mL5min69.6%AUCANP19.25ACV34.05GANPiStudyontheglycosylgalactoseacyclovirtripartedpre-drugnanoparticledeliverysystemHongHuayun(pharmaceutics)Supervisor:ZhuLiang(professor)ABSTRACTObjective:Toprepareacyclovirtripartedpre-drugandnanoparticle,thesurfacewasmodifiedbygalactosethencouldovercomethelimitedaccessofdrugtolivercellsandincreasedlivertarget.acyclovirsuccinimideactiveesterwaspreparedbyN-hydroxysuccinimideandSACVactivatedwithDCC,BSA-ACVwaspreparedbyBSAandacyclovirsuccinimideactiveester,it’sstabilityindifferentpHbuffersolutionwasevaluatedaswell;ThenmakeBSA-ACVintonanoparticleusingdesolvationpreocess.ToxicitytoHepG-2cellsandlivertargetinvivowerestudied.Methods:Theorthogonaldesignwasappliedtotheoptimizationofthesynthesis.TheeffectssuchastherateofNHSandSACV,therateofDCCandSACV,thereactiontime,temperatureandactivationratewereinvestigated.Theactivationratewas79.1%undertheoptimalcondition,ThesynthesisconditionsofBSA-ACVincludingBSAconcentrantion,pH,reactiontimeandmoleratiowereoptimizedinordertoreachhighcouplerate,TheconcentrationsofACVindifferentpHbuffersolutionweredeteminedbyHPLC,andthenindirectcalculatedtheconcentrationsofBSA-ACV,fromwhichcoulddeducethefunctions.Nanoparticlewaspreparedbydesolvationmethod,theinfluenceelementsincludingBSA-ACVconcentration,pH,alcoholconsumptionandtheamountofcuringagentwereevaluatedtoreceivethebestpropertiesofthemorphology,sizedistribution,Theoptimalpreparationelementswereconfirmedandcoupledwithglycosylonnanoparticlesurface.TheproductswereanalyzedandcharacterizedbySpectrophotometer,FTIR,TEM,LPA(laserparticleanalyzer).ThecytotoxicityofGANPoniihumanlivercancercellsHepG-2comparedwithANPandACVsolutiongroupwasstudiedusingMTTmethord,ThelivertargetofGANPandANPwereinvestigatedafterintraperitonealinjectioninmice,ComparedwithACVsolutioninjection.Result:TheactivatedSACVanalyzedbyFTIRhadthecharacteristicpeakofNHS,soitshowedtheproductwasacyclovirsuccinimideactiveester,andactivationratewas79.1%,BSA-ACVanalyzedbySpectrophotometer,FTIR,andDTA(MicrocumputerDifferentialThermalAnalyzers)indicatedthatBSAconnectedsomenumbersofACV,andthencouplerateofBSA-ACVmeasuredbyninhydrincolorationmethodreached24.1%.ThestudyofBSA-ACVstabilitysuggestedthatitdisplaypseudo-first-orderkineticsinbuffersolutions,andItdegratedfasterwhenPHincreased,andit’shalf-lifewas58.73hwhenpHwas2.0,but5.41hwhen9.7,AfteranlyzedbyTEMandLPA,Itindicatedthatnanoparticlepossessasmoothandroundappearanceandaveragediameterwas187nm,drugloadingwas4.2237%,Afterreleasestudyinvitroshowedthat56.19%ofACVcouldbereleasedfromGANPoveraperiodof48h.TheGANPgrouphadmostapparenthighrateofinhibitionreached96.5%,andIC50wasonly0.954mg/mL,farlessthantheIC50ofacyclovirsolutionwhichwas8.853mg/mL.Afterintraperitonealinjection,maximallyabout69.6%oftheinjucteddosewastakenupbytheliver,TheAUCinliverwasabout19.25timesofANPgroup,34.05TimesofACVsolutiongroup.Conclution:Theresultsofthestudyindicatedthatthepreparationofacyclovirforthetripartedpre-drugandthenpreparedfornanoparticlewhosesurfacecoupledwithglycosylasadrugdeliverysystems(TDDS),Whichcouldenhancetheuptakeofdrugsintoliver,wasaneffectivestrategyandanewapproachforthetreatmentofhepatitisB.Keywords:galactose;acyclovir;tripartedpre-drug;livertargeting1HepatitisBVirus,HBVHBsAg1HBVHBVHBVHBsAg[1]HBsAgHBsAgDNADNADNApolymeraseDNAPHBcAg[2]HBsAg30%40%[3,4]50[5]HIV[6][7](AcyclovirACVC8H11N503)[8]2ΗΝΝΝΝΟNΟΟΗHH1982IIIEBEpsteinBarr[9]DNADNADNA1982Weller[10]Smith[11][12][13,14][15][16,17]2070Narty3011000nm3[18]RES0.13µm730µm50nmalbumin[1920]serumalbumineggalbumin[21]100[22]ASGPR20%pH9MTT45AcyclovirACVC8H11N5031SOCl2COCl2DMFDMAP246--135-/TEAPPh3/CCl42[23,24,25]3[26]246-PfOHN-HOSuN-HOBt4DCC[27]6H3CCOH2CCOH2CH2CCH(CH3)NOOOCROMixedCarbonicAnhydrideActiveEsterRCOOCNHNO-Acylisoucea(DCC)O-DCC1DCCNHSDCCBoissonnas[29]NHSSACVDCCSACVSACVNHNNNONH2OOHsuccinconhydrideDMF/NEt3NHNNNONH2OOOHOONOOOHDMF/DCCNHNNNONH2OOOOONOO2711.11TU1800PCNicolet470FT-IRBS224SSartoriusZK-82A1.22AcyclovirACV99.8%N-NHSCPDCCCPCPCP22.1CollaL[30]1.1125g5mmol1.0g0.71mLNN-DMF6021h40mL2mol/LHclpH2SACV82.2SACVHSSACVNN-DMFN-NHSDCCDMF2.2.1XHX=CNO[31]12mg200mgKBr2µm510107pa3433428cm-1NH21748cm-1COOR1695cm-1COOH43452cm-1NH21734cm-11616cm-1C=O1831cm-11792cm-1C=O1206cm-1O9342.2.2NHSMiromTaliaWilchekMier[32]19820.1mol/LN-260nmNHSNHS10HNNNNOH2NOOOOONH4OHHNNNNOH2NOOOOHONOOOHNOO56NHS10.1mol/L100mL20.5mmol/LNHS3260nm3NHS35[NHS]/mmol/L/mLNHS/mL/mLA2600300.500.530.10.40.126130.20.30.2341.530.30.20.335230.40.10.4342.530.500.54011y=0.206x+0.025R2=0.999800.10.20.30.40.50.60123CA7NHSy0.206x0.025R20.99980.5mmol/L3mL0.5mL260nm()%1005.05.05.335206.0025.0××××−=Aα2.3NHSSACVN-DCCNHSSACVDCCSACVN-NHS2.3.12.22h124h6h8h10h4h%12422463.136474833.551030.8%20304050607002468101