[生物医药论文精品a]羟基喜树碱转铁蛋白隐形纳米泡囊的制备和体内、外评价

1羟基喜树碱转铁蛋白隐形纳米泡囊的制备和体内、外评价洪鸣凰，裴元英A1A2A0A3A4A6A4A5A8A9A7200032A11摘要目的以羟基喜树碱（HYDROXYCAMPTOTHECIN,HCPT）为模型药物，制备转铁蛋白（TRANSFERRIN,TF）修饰隐形纳米泡囊，探索聚乙二醇（POLYETHYLENEGLYCOL，PEG）修饰的隐形化效果与TF修饰的主动靶向相结合，有效递送药物到达肿瘤部位的可行性。方法以人工合成的聚乙二醇化聚十六烷基氰基丙烯酸酯（PEGPHDCA）为表面修饰材料，共价连接氧化的TF，采用薄膜水化超声分散法制备载HCPT隐形纳米泡囊（TFPEGNS），考察粒径、包封率、体外释放、细胞毒性、细胞摄取和细胞内分布，大鼠药动学和S180荷瘤小鼠体内组织分布，抗肿瘤疗效。结果TFPEGNS粒径为116NM，包封率为9300038，体外释放48H达90。体外KB细胞对TFPEGNS的摄取呈G10628G8999G5242与G7114G19400G1393G17194性，G1000G3324G1314G9213G7477G1226G6122G13785G17819G18339TF的G4396G3324G991，摄取G2475到G6245制。与G5078G2818G8892G4568G2070、G7234G17902泡囊和PEG修饰泡囊（PEGNS）相G8616，TFPEGNS对KB、K562、S180G989G12193肿瘤细胞毒性G7380G5390G727胞内药物摄取G18339，G4600G1866G7171细胞G7692内G7380G3822G727肿瘤部位药G7114G7366G13459G991面G12227G7380大G727抗肿瘤G8975性G7380G5390，对S180荷瘤小鼠的肿瘤G6245制率达71。结论G6164制备的转铁蛋白修饰隐形纳米泡囊可G1328为有效的载体递送HCPTG14279肿瘤细胞G2469G6393疗效。关键词G726转铁蛋白G727羟基喜树碱G727G19762G12175G4388表面G8975性G2070泡囊G727聚乙二醇G727肿瘤靶向PREPERATIONOFTRANSFERRINMODIFIEDHYDROXYCAMPTOTHECINLOADEDPEGYLATEDNIOSOMESANDITSEVALUATIONINVITROANDINVIVOHONGMINGHUANG,PEIYUANYINGA10SCHOOLOFPHARMACY,FUDANUNIVERSITY,SHANGHAI200032,CHINAABSTRACTOBJECTIVETOEXPLOITTHEPOSSIBILITYOFCOMBINATIONOFTHESTEALTHACTIONBYPOLYETHYLENEGLYCOLCYANOACRYLATECOHEXADECYLCYANOACRYLATEPEGPHDCAMODIFIEDNIOSOMESANDACTIVETARGETINGFUNCTIONOFTRANSFERRINTFBYA12A13A14A15A16A17A18A19A20A21973A22A23A242007CB935802A25A26A16A17A27A28A29A30A18A31A2430572266A25A32A33A22A23A34A35A36A37A38A39A40,A41,A42A43A44A45A46A47A48A49A50,A42A43A51A52A53TEL02154237186EMAILYYPEISHMUEDUCN2TRANSFERRINRECEPTORMEDIATEDENDOCYTOSISTOPROMOTEDRUGDELIVERYTOSOLIDTUMORFOLLOWINGINTRAVENOUSADMINISTRATIONWITHHYDROXYCAMPTOTHECINHCPTASMODELDRUGMETHODSHCPTLOADEDPEGNIOSOMESPEGNSWEREPREPAREDBYTHINFILMHYDRATIONANDULTRASOUNDMETHODTFWASCOUPLEDTOTERMINALAMINOGROUPOFPEGTOPRODUCETHEACTIVETARGETINGVESICLESTFPEGNSTHEPARTICLESIZEDISTRIBUTION,ENTRAPMENTRATE,INVITRORELEASE,CYTOTOXICITY,CELLUPTAKE,INTRACELLULARDRUGDISTRIBUTION,INVIVOPHARMACOKINETICSINRATS,TISSUEDISTRIBUTIONANDANTITUMOREFFECTINS180TUMORMICEMODELWEREDETERMINEDRESULTSTHEPREPAREDTFPEGNSWITHAVERAGEDIAMETERSOF116NMSHOWEDSUSTAINEDRELEASEPROFILEINVITROTHEUPTAKEOFTFPEGNSINTOKBCELLSWASCONCENTRATIONANDTIMEDEPENDENT,WHICHCOULDBEINHIBITEDBYLOWTEMPERATUREANDFREETFCOMPAREDWITHHCPTINJECTION,NONSTEALTHNIOSOMESANDPEGNS,TFPEGNSDEMONSTRATEDTHESTRONGESTCYTOTOXICITYTOTHREECARCINOMATOUSCELLLINESKB,K562ANDS180CELLS,THEGREATESTINTRACULLARUPTAKEESPECIALLYINNUCLEI,THEHIGHESTTUMORCONCENTRATIONANDLARGESTAREAUNDERTHEINTRATUMORALHYDROXYCAMPTOTHECINCONCENTRATIONCURVE,ASWELLASTHEMOSTPOWERFULANTITUMORACTIVITYWITHTHEINHIBITIONRATEOF71AGAINSTS180TUMORINMICECONCLUSIONTHETRANSFERRINMODIFIEDPEGYLATEDNIOSOMESCOULDBEONEOFTHEPROMISINGSOLUTIONSTODELIVERYANTITUMORDRUGSTOTUMORKEYWORDSTRANSFERRINHYDROXYCAMPTOTHECINNIOSOMESPOLYETHYLENEGLYCOLTUMORTARGETINGG19762G12175G4388表面G8975性G2070泡囊NONIONICSURFACTANTVESICLES，NIOSOMESG7171G11013合成的G19762G12175G4388表面G8975性G2070G3324水性G1183G17148G1025组成G2345G4472G6122G3822G4472封G19393G2464G4630结G7512的有G5219组织聚G19610体，G12628G12228泡囊。G11013G19762G12175G4388表面G8975性G2070G7379G1207G11979G14038G13792形成的泡囊相G8616G1122G14038G17148体，G1867有成分G11842G4462、结G7512G12295G4462、G7143G1122G1457G4396、成G7424G1314、G7092毒性G2462G993G14403G2465G5224G12573G1260G9869。G9994G13792，G1328为G14026体载药G13007G13491，泡囊G19757G14045G8892G4568G2530G1262G15999G2345G7692G5052G3136细胞G13007G13491（MPS）摄取G13792G5567G17907G9177G19512。采用G1158水性G20652分G4388材料,G3926聚乙二醇POLYETHYLENEGLYCOL,PEG修饰制备隐形纳米载药体G13007,可以G18003G1825MPS的G2546G3136,G5322G19283药物G3324G15892G9094G5502G10627G13007G13491G1025的G9394G11053G7114G19400,G17839G13792G17902G17819EPR效G5224G4512G19610G1122肿瘤组织。用G1867有肿瘤靶向性的G3848基修饰泡囊，可以G17839G980G8505G6564G20652抗肿瘤药物的疗效。转铁蛋白G2475体G3324G3822G12193肿瘤细胞表面G20652表达，G1363G1866G1328为靶G98693G1183G4560药物转G17828到G3698值细胞内成为可能1,2。G7424实验G4472前期成功合成了转铁蛋白聚乙二醇肿瘤坏死因G4388共轭物TFPEGTNF3，G1866与肿瘤细胞表面TFR的结合常数与游G12175TF相似。共轭物G15892浆半衰期较原药明显G5322G19283，肿瘤蓄G12227G18339G3698加，抗癌G8975性分别G7171TNF和PEGTNF的53和18倍。近年来，已有TF药物共轭物4、TFG14038G17148体5、纳米粒6、树突状聚合物7G12573研究，但未见转铁蛋白修饰G19762G12175G4388表面G8975性G2070泡囊的报G4560。羟基喜树碱（HYDROXYCAMPTOTHECIN,HCPT）G7171喜树G1025分G12175G6564取的同类抗肿瘤G2345体G1025抗癌G1328用G7380G5390的微G18339生物碱。临床主要G17828用G1122消化G13007G13491癌症G727对口腔、颈面部癌、G3848颈部圆柱型腺癌G2462膀胱癌也有较好疗效。但G7171，HCPTG4396G3324半衰期短（530MIN左右），G14038溶性和水溶性均差，G12295G4462性差以G2462毒副G1328用大G12573缺G9869，G1866临床G5224用G2475到G980G4462的限制。G7424课题组前期工G13288以SPAN60和胆固醇为载体，聚乙二醇化聚十六烷基氰基丙烯酸酯MEPEGPHDCA为膜表面修饰材料，制备了HCPT隐形纳米泡囊。结果表明粒径G332490NM左右MEPEG5000PHDCA载药纳米泡囊G1867有G7380佳的肿瘤靶向、规G18003肝脾G1328用和G6245癌G1328用。G3324此基础上，G7424文以TF为靶向G3848基，制备了载HCPT的主动靶向隐形纳米泡囊，G17839行体外药物释放、细胞毒以G2462亚细胞器（包括细胞G17148和细胞G7692）药物经G7114分布评价，大鼠体内药动学、荷瘤小鼠组织分布、抗肿瘤疗效，并与未修饰泡囊和HCPTG8892G4568G2070G17839行G8616较，来探索以泡囊为载体，用PEG修饰的隐形化效果与TF修饰的主动靶向相结合，有效递送药物到达肿瘤部位的可行性。1材料与仪器11材料T叔丁氧羰基氨基羟基聚乙二醇BOCNHPEGOH,MW5,000（德国IRISBIOTECHGMBH公司）G727羟基喜树碱（上海骏杰生物技术有限公司）G727羟基喜树碱G8892G4568G9094（湖北黄石药业有限公司）G727NH2PEGPHDCA、MEPEGPHDCA与PHDCA（G7424实验G4472自制）G727司盘60、胆固醇（G1025国医药（G19610团）上海化学试G2070公司）G727甲氧基聚乙二醇MEPEG,MW5,000、HOLOTRANSFERRIN、苯甲基磺酰氟（美国SIGMA公司）G727人转铁蛋白ELISAG4462G18339试G2070盒美国BETHYL公司G727SEPHAROSECL4B（瑞典PHARMACIA公司G727胎牛G15892G9177（杭州四季青公司）G727RPMI1640、胰蛋白酶（美国GIBCO公司）G727MTT（美国AMRESCO公司）G727BCA蛋白G4462G18339试G2070盒（美4国ROCKFORD公司）G727G1866G1325试G2070均为分G7524G13443。12细胞人G5942性G20647性白G15892G11161K562细胞、人口腔表G11394G7691癌KB细胞、小鼠G14418G8675G14157水瘤S180细胞、G8503常人肝L02细胞（G1025G12197G19510上海生物细胞学研究G6164）13动物G19608性WISTAR大鼠20020G和G7130明小鼠202G（G3809G7098大学实验动物部）14G1214器UV2401PCG13055外分G1821G1821G5242G1214G7097G7424SHIMADZU公司G727NICOMPTM380ZLS粒G5242/ZETAG11017位G8991G4462G1214美国NICOMP公司G727MERCURYPLUS400MHZG7692G11925共G6403G1214（美国VARIAN公司）G727LC10ATSPD10AG20652效G9094相G14406G16901G1214（G7097G7424SHIMADZU公司）G7271100SERIESG14651G1821G20652效G9094相G14406G16901G1214（美国AGILENT公司）G727ELX800酶G7643G1214（美国BIOTEK公司）。2方法21HCPT纳米泡囊的制备G2462表G5461G3324G7424实验G4472前期基础上8，采用薄膜水化超声分散法制备未修饰的PHDCA泡囊（NNS）和PEG修饰泡囊（PEGNS）。主动靶向泡囊的制备G2454考文G104989，并加以G6925G17839。G20330G1820以NH2PEGPHDCA为修饰材料，制备隐形纳米泡囊（NH2PEGNS）。G2490G4570TF100MG溶G1122HEPESG13543G1926G9094（HBS，150MMNACL，20MMHEPES，PH74,NAIO44MG溶G1122G18271酸G19060G13543G1926G9094（30MM，PH5），G9163合G2530G1122G7275G3800、G1924G9032G7477G1226G991G2465G522490MIN，经SEPHADEXG25PD10柱、HBSG8939G14085，G1122280NMG3800G11429G8991G5483到氧化转铁蛋白。超G9400G12175G5527G12661（MWCO3,000）G12175G552710MING8999G13565G2530，G17817G17907加G1849G14279NH2PEGNS（G6365G10043G6717G4584G8616PEGG726TF2G7261），G4472G9213G6617G6304G2465G522412HG2530，以HBSG8939G14085G17819SEPHAROSECL4BG1969G14026柱，G1122280NMG3800G11429G8991，G19512G2447未连接蛋白，G2375G5483HCPT的主动靶向隐形纳米泡囊（TFPEGNS）。采用TFELISAG4462G18339试G2070盒G8991G4462G5647TFG1022数，G8616G8990法G8991G4462泡囊G1022数，G1016G13785G1055G8616G2375为G5191均G8611G1022泡囊表面的TF连接G1022数10。G7693G6466实验G4472原有G7053法29G8991G4462TFPEGNS、PEGNS和NNS的粒径、ZETAG11017位、包封率、载药G18339。22体外释放G5242实验采用转G12738法考察HCPT溶G9094、TFPEGNS、PEGNS和NSG3324G12573G9195PBS（PH574）的释放G5242。23体外细胞毒考察G993同制G2070，包括HCPTG8892G4568G2070、TFPEGNS、PEGNS以G2462NNS的细胞毒性。K562、KB、S180、L02细胞以1104G1022/G4392接G12193G112296G4392G7507，37G263G3533G187124HG2530，分别加G1849G993同G8999G5242的G2520G12193制G2070（3000，2500，1250，650，300，150，075，050GG934ML1），G13499G13505G3533G187124HG2530，采用MTT法G8991G5483IC50。24细胞摄取KB细胞以1105G1022/G4392接G12193G112224G4392G3533G1871G7507，37G263G3533G187124HG2530，分别加G1849G8999G5242为15GG934ML1的HCPTG8892G4568G2070、TFPEGNS、PEGNS以G2462NNSG252003ML，G13499G13505G4425G13958，G1122G993同G7114G19400G9869（025，05，1，2，4H）取G1998。G1924G11979酸G11428G13543G1926G9094（PBS，PH74）G8939G9080G989G17953，消化，3000RG934MIN1G12175G55275MING6922G19610细胞，G1889用PBS3MLG8939G9080G989G8437，1TRITONX10004MLG4425G13958G17819G3824。G12175G5527G19512G2447细胞G11874G10267，采用G14651G1821HPLCG8991G4462细胞G16022G16311G9094G1025的药G18339，BCA试G2070盒G8991G4462细胞蛋白G8999G5242，摄取G12255G5242表G12046为HCPTNG/蛋白G18339MG。同法考察4G263G4425G13958，G6122G13785G17819G18339TFG4396G3324，G6122G13785G993同HCPTG8999G524205,1,25,5,15,30GG934ML1G112237G263G4425G139581HG7477G1226G991的细胞摄取G12255G5242。25亚细胞器药物分布G2454考文G1049811并加以G6925G17839。KB细胞以1106G1022/G4392接G12193G1122100MMG3533G1871G11411，37G263G3533G187112HG2530，分别加G1849G8999G5242为100GG934ML1的HCPTG8892G4568G2070、TFPEGNS、PEGNS以G2462NSG25201ML，G13499G13505G4425G13958，G1122G993同G7114G19400G9869（05，1，2，4，8，12H）取G1998，PBSG8939G9080G989G17953，消化，3000RG934MIN1G12175G55275MING6922G19610细胞，G1889用PBS3MLG8939G9080G989G8437，G8797G9108用TM2G13543G1926G9094（001MTRISHCL，PH74，0002MMGCL2，00005M苯甲基磺酰氟）300LG18337G5760，G4472G9213G991G19757G85021MING2530，G1924G90325MIN，G13499G13792加G1849TRITONX10015L，G1924G90325MIN，G11224G263，800RG934MIN1G12175G552710MIN，上G9177G9094用G1122G7828G8991细胞G17148G1025药物G2559G18339，G8797G9108用TM2G13543G1926G90941MLG18337G5760G2530用G1122G7828G8991细胞G7692G1025药物G2559G18339。细胞G17148和细胞G7692蛋白G2559G18339采用BCA试G2070盒G8991G4462。26大鼠体内G15892浆药动学24G2494G19608性WISTAR大鼠G19555G7438分为四组,G6365组别G4626G19757G14045分别G8892G4568G5078G2818HCPTG8892G4568G2070、TFPEGNS、PEGNS以G2462NS,G2070G18339为08MGG934KG1。G13485药G2530分别G11220083,025,05,1,2,4,6,8,12,24HG1122G11536G5225取G15892，采用G14651G1821HPLCG8991G4462G15892药G8999G5242。627S180荷瘤小鼠体内组织分布和抗肿瘤药效学采用IODOGEN法制备125IHCPT。S180荷瘤小鼠G19555G7438分为四组，G6365组别G4626G19757G14045分别G8892G4568G5078G2818HCPTG8892G4568G2070、TFPEGNS、PEGNS以G2462NS,G2070G18339为1MGG934KG1125CI。G13485药G2530分别G112205,2,4,8,12,24H取G15892G2530G3800死，取肿瘤、G5527、肝、脾、G13966、G13970、G14053，G12228G18337，G8991G4462放G4568性G16757数，G16757G12651相对摄取率ID/G。G7130明小鼠接G12193S180瘤24小G7114G2530G19555G7438分为5组，G19464性对G10043组G13485G1116生G10714G11428水,G1866G13254组HCPT制G2070G63651MGG934KG1连G13505G19757G14045G13485药7G3837。停药24HG2530G3800死动物，G12228体G18337、瘤G18337，求G1998肿瘤G6245制率。并以4G3822聚甲醛固G4462,石蜡包埋,切G10267,HE染G14406,G1821学显微镜G991G17839行肿瘤组织G11161G10714组织学观察。28G13491G16757分G7524抗肿瘤药效学结果采用ANOVAG7828验，G1866G1325数G6466分G7524采用TG7828验判断差异的显著性，结果以SX表G12046，PPEGNSNNSG8892G4568G2070G727G13792对G1122G8503常细胞，毒性差异G993明显。表2HCPT制剂对肿瘤细胞和正常细胞毒性IC50值N3TAB2THEIC50VALUESOFHCPTPREPARATIONSAGAINSTCARCINOMATOUSANDNORMALCELLLINESN3INJECTIONUG/MLNNSUG/MLPEGNSUG/MLTFPEGNSUG/MLK5625384740623027242993260533细胞摄取KB细胞对泡囊的摄取呈G10628G8999G5242G1393G17194性，并G19555G4425G13958G7114G19400的G5322G19283，摄取G18339G3698加8FIG2A,B。与PEGNS相G8616，TFPEGNS的摄取G17907G5242G5567，G12255G5242大。与37C相G8616，4CG4425G13958G7114细胞对TFPEGNS的摄取G12255G5242有显著性G991降，仅为4C的2672（P005。图2KB细胞对TFPEGNS和PEGNS的摄取A,G993同G8999G524237COR4CG7477G1226G9919G4425G139581HB,固G4462G8999G524215G/ML，37CG7477G1226G991G4425G13958G993同G7114G19400C,固G4462G8999G524215G/ML，游G12175TF1MG/MLG4396G3324G61224CG4425G139581H。N3,P脾肝G13966G13970G5527G14053。12HG7114，G7092论与G1866他制G2070G6122G7171与G1866他脏器相G8616，TFPEGNS组G3324肿瘤的药物G8999G5242也G7171G7380G20652。图5C显G12046了G993同HCPT制G2070G3324肿瘤部位的药G7114G7366G13459。TFPEGNS组峰G8999G5242G7380G20652，分别为G8892G4568G2070、NNS和PEGNS的400,288和151倍G727药G7114G7366G13459G991面G12227G7380大，分别为G8892G4568G2070、NNS和PEGNS的1697,332和145倍。G4570药动学G2454数G17839G980G8505G3800G10714，采用相对摄取率（RE）和峰G8999G5242G8616（CE）来G17839行肿瘤靶向性评价（表4），G1866规律为G726TFPEGNSPEGNSNNS。表3HCPT泡囊与注射剂的相对摄取率（RE）和峰浓度比（CE）N3TAB3THERELATIVETUMORTISSUEEXPOSURESREANDTHERATIOSOFHCPTPEAKCONCENTRATIONSCEINTUMOROFNIOSOMESCOMPAREDWITHHCPTINJECTIONN3NNSPEGNSTFPEGNSRE51111731697C5HCPTG8892G4568G2070,NNS,PEGNS和TFPEGNSG13485药4HA和12HB组织分布C,肿瘤部位的药物G8999G5242G7114G19400G7366G13459N3,SXFIG5TISSUEDISTRIBUTIONSOFHCPTINJECTIONANDNNS,PEGNS,TFPEGNSAT4HOURSAAND12HOURSBAFTERADMINISTRATIONC,HCPTLEVELVSTIMECURVESINS180TUMORSOFMICEFOLLOWINGINTRAVENOUSADMINISTRATIONOFHCPTPREPARATIONSN3,SX37体内抗肿瘤药效泡囊的体内抗肿瘤药效试验结果G3926图6，表4G6164G12046，疗G12255结束G2530，G19464性对G10043组的G5191均瘤G18337为068G，G13792TFPEGNS组仅为020004G，显著轻G1122G8892G4568G2070组04813G,P001,NNS组044G,P001和PEGNS组037G,P001。肿瘤G6245制率达71，分别为PEGNS，NNS和G8892G4568G2070的155,200和240倍，疗效有显著性G6564G20652，未见明显的毒副G1328用。图6HCPTG8892G4568G2070,NNS,PEGNS和TFPEGNS治疗G2530肿瘤大小FIG6TUMORGROWTHAFTERSYSTEMICAPPLICATIONOFHCPTINJECTION,NNS,PEGNSANDTFPEGNS表4HCPT注射剂和泡囊的S180荷瘤小鼠体内抗肿瘤药效TAB4ANTITUMOREFFECTSOFHCPTANDHCPTLOADEDNIOSOMESAGAINSTSARCOMA180TUMORINMICEDOSENUMBEROFANIMALSBODYWEIGHTGTUMORWEIGHTGINHIBITIONMG/KGINITIALFINALINITIALFINALMEANSDSALINE20202182849068014HCPT1710102149281804800329NNS1710102142283204400235PEGNS171010216284203700346TFPEGNS1710102151280902000471肿瘤组织G11161G10714组织观察显G12046（图7）G726生G10714G11428水G19464性对G10043组肿瘤组织G1025癌细胞丰G4512,G7692大深染,肿瘤细胞生G19283旺盛,有较G3822G7692分G16022像,呈卵圆形、梭形、G989角形G246214G993规则形,呈淡蓝G14406,大G3822数细胞内有1G1022G7692,部分细胞内有2G1022G6122以上的G7692，胞浆较少。采用HCPT制G2070治疗G2530，肿瘤组织有着G993同G12255G5242的坏死。G4600G1866G7171TFPEGNS组的肿瘤组织大G10267坏死,G8503常结G7512消失。癌细胞数G18339减少，胞浆较丰G4512，呈淡红G14406，可见G7692G8999G13565、G7692G11874G16022，G8999G13565的染G14406G17148可围绕G1122G7692周边G6122形成新月体结G7512。图7生G10714G11428水A,HCPTINJECTIONB,NNSC,PEGNSD和TFPEGNSE治疗G2530肿瘤部位G11161G10714切G10267FIG7HESECTIONSOFTUMORAFTERSYSTEMICAPPLICATIONOFSALINEA,HCPTINJECTIONB,NNSC,PEGNSDANDTFPEGNSE4讨论G3324治疗疾G11161的G17819G12255G1025，为了G6564G20652药物疗效，降G1314G1866毒副G1328用和减少药源性疾G11161，G4570药物G4462向G8999G19610G1122靶器官、靶组织、靶细胞的靶向G13485药G13007G13491已成为G10628G1207药G2070学研究的热G9869G1055G980，G1866关键G3324G1122寻找合适的靶G3848。转铁蛋白G1328为内源性蛋白G1867有G7092毒、G7092G1825疫原性、可生物降G16311G12573特G9869，因G13792备G2475青睐12。转铁蛋白连接到微粒表面常见的G7053法有戊二醛交联法、转铁蛋白巯基化以G2462转铁蛋白氧化G12573，前G1016G12193G7053法分别以转铁蛋白分G4388上的氨基和二硫键为G2465G5224部位，G4396G3324着G2465G5224位G9869G993明G11842的问题。G7424文采用G20652碘酸G11428氧化法G1867有显著的G1260势，这G7171因为TFG17902G17819糖基化G1328用部位上的糖苷键与NH2PEGNSG2465G5224,已证明该糖基G993影响TF与TFR的结合。G13792G1000，伸G1998的糖基链G3698加TF同泡囊表面的距G12175，降G1314与G2475体结合的位阻，从G13792G1457证TF的结合G8975性损失G7380G1314。15与PEGNS，NNS相G8616，TFPEGNSG1867有G7380G5390的肿瘤细胞毒性。TFPEGNS、PEGNS的细胞摄取都能G15999G1314G9213G6164G6245制，证明泡囊的内G2546G7171G980G1022能G18339G1393G17194的G17819G12255。TFPEGNS的细胞结合（4C，G993内化）和蓄G1222737C,内化都要G20652G1122PEGNS。游G12175TF能显著G6245制TFPEGNS的摄取，证明TFPEGNSG7171G17902G17819G2475体G1183G4560的细胞内G2546G1849胞。G13792G1000，细胞内药物分布实验还G2469G10628TFPEGNS组细胞G7692G1025药物水G5191G7380G20652，这对G1122HCPT疗效的G2469G6393G7171很有利的，因为HCPTG7171G1328用G1122细胞G7692G1025的TOPOG13792G6245制肿瘤的。与体外细胞毒试验结果G980致，TF修饰G2530泡囊G3324S180荷瘤小鼠体内药效也有显著G6564G20652。G13485G1116较G1314G2070G18339（1MGG934KG1）情况G991，TFPEGNS的肿瘤G6245制率达71，分别为PEGNS，NNS和G8892G4568G2070的155,200和240倍。疗效的G6564G20652可以归结为以G991几G7053面原因。G20330G1820，PEGPHDCA的修饰起到了隐形化效果，从G13792G5322G19283泡囊的G15892G9094G5502G10627，有利G1122G17902G17819EPR效G5224G4512G19610G1122肿瘤组织。第二，TF修饰泡囊G4570HCPTG4462向输送到肿瘤部位。第G989，TF修饰泡囊能有效地G3698加肿瘤细胞G7692内药物G18339，有利G1122抗肿瘤药物疗效的G2469G6393。G7380G2530，泡囊的包封能有效地G6564G20652药物的G12295G4462性，有利G1122G1866G8975性的G1457持。G6164制备的TFPEGNSG1867有G13543释G1328用，能G3698加药物的G7692内摄取G18339，G3698G5390细胞毒性，G5322G19283G13007G13491G5502G10627G7114G19400，G1363药物G4512G19610G1122肿瘤，从G13792G6564G20652药物的抗肿瘤疗效。因此，G17902G17819G4570TF共价连接到PEG修饰的泡囊G6164G5483的主动靶向泡囊G7171G980G12193较G1867潜力的抗肿瘤药物载体。当G9994，载药G13007G13491内G2546G7438制G2462胞内转G17828途径仍需G17839G980G8505研究，G1866他因素，G3926膜流动性和表面靶G3848密G5242的影响也值G5483G17839G980G8505探讨。REFERENCES1MARUYAMAK,ISHIDAO,KASAOKAS,ETALINTRACELLULARTARGETINGOFSODIUMMERCAPTOUNDECAHYDRODODECABORATEBSHTOSOLIDTUMORSBYTRANSFERRINPEGLIPOSOMES,FORBORONNEUTRONCAPTURETHERAPYBNCTJCONTROLRELEASE2004981952072KURSAM,WALKERGF,ROESSLERV,ETALNOVELSHIELDEDTRANSFERRINPOLYETHYLENEGLYCOLPOLYETHYLENIMINE/DNACOMPLEXESFORSYSTEMICTUMORTARGETEDGENETRANSFERBIOCONJUGCHEM200