靶向整合素_v_3受体的新型RGD肽二聚体的_131_I标记与生物活性的初步

:(973,2006GB705705)(,985-2-056)(0707)SupportedbytheMajorStateBasicResearchProgramofChina(973Program,2006GB705705),theSpecialFundforPromotionofEducation,MinistryofEducationP.R.C.(985-2-056),andtheKeyLaboratoryofRadiopharma-ceuticalsofMinistryofEducation,CollegeofChemistry,BeijingNormalUniversity(0707)Correspondingauthorse-mai,[e-mail],[e-mail]:2011-3-495530:技术方法靶向整合素v3受体的新型RGD肽二聚体的131I标记与生物活性的初步评价张春丽1,2,王荣福2,张丽2,郭凤琴2,李玲2,康磊2,闫平2,杨铭1(1.,100191;2.,100034)[]目的:--(Arg-Gly-Asp,RGD)131I,,方法:c(RGDfV),cRGD[c(RGD)2],ChT131I,c(RGD)2;,131I-c(RGD)2结果:131Ic(RGD)2(76.352.33)%,SephadexG10(95.203.25)%c(RGD)2Ka(0.1370.057)109/mol/L,lgP/-1.628,131I-c(RGD)21h3h5h24h(0.670.13)%ID/g(0.420.08)%ID/g(0.510.11)%ID/g(0.180.02)%ID/g,,(T/NT),24h,(T/M)(T/B)4.421.702.270.45131I-c(RGD)2,1h2.100.60;131I-c(RGD)2,3h结论:c(RGD)2ChT131I,,131I-c(RGD)2,[];;;v3[]R977.6[]A[]1671-167X(2011)02-0295-06do:i10.3969/.jissn.1671-167X.2011.02.026131IlabelingandbioactivityevaluationofanovelRGDdimertargetedtointegrinv3receptorZHANGChun-li1,2,WANGRong-fu2,ZHANGLi2,GUOFeng-qin2,LILing2,KANGLei2,YANPing2,YANGMing1(1.StateKeyLaboratoryofNaturalandBiomimeticDrugs,PekingUniversitySchoolPharmaceuticalSciencesCenter,Beijing100191,China;2.DepartmentofNuclearMedicine,PekingUniversityFirstHospita,lBeijing100034,China)ABSTRACTObjective:Tostudythe131Ilabeling,tumortargetingproperty,biodistrubutionandimagingofanoveldisulfidebridgedArg-Gly-Asp(RGD)peptidedimerandinvestigateitspossibilityfortumorangiogenesisimaging.Methods:AdisulfidebridgedcRGDpeptidedimer[c(RGD)2]wasdesignedac-cordingtothepublishedresultsofstructure-activityrelationshipstudyofc(RGDfV)andmodifiedwithhydrophilicaminoacid.Theaffinityofthepeptidetov3receptorwasdeterminedbybindingassay.Thepeptidewaslabeledwith131IbyChTmethod.Thestabilityandlgn-octano-lwaterpartitioncoefficientofthelabeledpeptideweretested.Thebiodistribution,inhibitionwithunlabeledpeptideandimagingwereperformedbyinjectingthelabeledpeptideintothemicebearingmelomaB16.Results:The131Ila-belingefficiencyandradiochemicalpuritywere(76.352.33)%and(95.203.25)%,respectively.Kawas(0.1370.057)109/mol/L,andlgPoctanol/waterwas-1.628.Thetumoruptakeofthelabeledpeptideat1h,3h,5hand24hwere(0.670.13)%ID/g,(0.420.08)%ID/g,(0.510.11)%ID/gand(0.180.02)%ID/g,respectively.Thetumorclearancewaslow,sotheT/NTincreasedwithtime.TheT/MandT/Bwere4.421.70and2.270.45,respectively,24h295()JOURNALOFPEKINGUNIVERSITY(HEALTHSCIENCES)Vo.l43No.2Apr.2011pos-tinjection.Thehepaticuptakeofthelabeledpeptidewaslow.Thetumor-to-liverratiowas2.100.60at1hpos-tinjection,coinjectionofc(RGD)2with131I-c(RGD)2resultedinasignificantdecreaseintumoruptake.Intotalbodyimages,onlykidneysweredistinctivewhereastumorwasclearinchestimages3hpos-tinjection.Conclusion:c(RGD)2canbesuccessfullylabeledwith131IbyChTmethod.Thelabeledpeptidecanbespecificallycombinedwithtumor,whichsuggestsitspossibilityintumoran-giogenesisimaging.Itslowliveruptakeprovidesanadvantagefortumorimaging.KEYWORDSOligopeptides;Dimerization;Iodineradioisotopes;IntegrinalphaVbeta3,v3,,[1]--(Arg-Gly-Asp,RGD)v3,RGD[2-4],RGD,,,RGD,,RGD,131I,,11.1:HANGPINGFA1104,;FT609,;RM905,;SPECTGE-MRP,GE:RGD,c(RGD)2,1,HPLC95%,;-T,,;Na131INa125I,;,1c(RGD)2Figure1Aminoacidsequenceofc(RGD)2:B16,U87,:(SPF),,15~17g,,1.2131I(125I)T(ChT)0.5g/Lc(RGD)20.5mol/LpH7.4,131I-NaI(125I-NaI),ChT0.9g/L,2min,SephadexG10131I(125I),1,//=5/1/2(),131I(125I)-c(RGD)2Rf=0~0.1,131I(125I)Rf=0.4~0.60.9~1.0131I-c(RGD)2437,1.3c(RGD)2v3U8710%()DMEM,2,0.25%()5105/100L24EP100L,,0nmol/L0.125nmol/L0.25nmol/L0.5nmol/L1nmol/L2nmol/L4nmol/LcRGD100L,,1105nmol/Lc(RGD)2100L,2105/min125I-c(RGD)250L,10%pH7.4Tris200L296()JOURNALOFPEKINGUNIVERSITY(HEALTHSCIENCES)Vo.l43No.2Apr.20113,42h,(T),,100LPBS2,,200LPBS,(B)Scatchard(B/FB,F,T-B),,Ka1.410L131I-c(RGD)21mL,1mL,1min,3000r/min3min,,100L,lgP/=lg(cts/cts),ctscts100L100L1.5B1610%RPMI-1640,,107/100L75%(),B16,1071.0cm131I-c(RGD)21.6255,5,4185kBq(5Ci)/0.17g131I-c(RGD)2100L,1h3h5h24h1;1,185kBq(5Ci)/0.17g131I-c(RGD)240gc(RGD)2,3h,,,(%ID/g,/100%)(T/NT,/)1.7131I-c(RGD)2131I,30.1%()131I2,7.4MBq(200Ci)131I-c(RGD)2,1h3h5h24h():256256,140keV,2,100K1.8SPSS16.0,tP0.0522.1131I131Ic(RGD)2(76352.33)%,SephadexG10,(95.203.25)%125I-c(RGD)287.25%99.01%131I-c(RGD)243723,,131I-c(RGD)241d376h90%131I-c(RGD)2lgP/-1.628,297,v3RGD131I2.2c(RGD)2v3c(RGD)2v3Ka(0.1370.057)109/mol/L2.3131I-c(RGD)2B161,T/NT4,c(RGD)2,,,1(0.670.13)%ID/g,,131I-c(RGD)2,T/NT(4)5h,;,T/NT1.4,24hT/NT25h24h,(T/M)4.190.314.421.70,(T/B)1.090.392.270.45,3h3ht,t=4.392,P0.05,4T/NTFigure4T/NTatdifferenttimepost-injection1131I-c(RGD)2[(xs)%ID/g,n=5]Table1Biodistributionof131I-c(RGD)2inmicebearingB16meloma[(xs)%ID/g,n=5]Tissues1h3h5h24h3h(Inhibitiongroup)Bladder0.590.320.360.120.710.360.240.000.580.26Stomach1.880.182.110.663.111.160.210.061.960.74Smallintestine0.410.560.330.150.340.100.070.020.330.11Kidney31.61.9219.36.1719.27.063.691.5522.94.46Liver0.320.040.230.060.220.040.060.000.230.06Lung0.720.160.520.250.360.050.090.020.390.08M