蛋白多肽类药物的药代动力学分析方法的研究进展

　,＊(,　100850):蛋白多肽类药物的药代动力学与传统药物不同,目标蛋白多肽给药量小,而各种内源性蛋白含量很高,这种干扰使准确测量目标分子非常困难。此类研究必须以建立灵敏、专一、准确的测定方法为前提。本文综述了近年来研究蛋白多肽类药物的药代动力学各种分析方法,包括生物检定法、同位素标记示踪法、免疫分析法、液相色谱、气相色谱、毛细管电泳、质谱、核磁共振、液相色谱-质谱(LC-MS)和毛细管电泳-质谱(CE-MS)联用技术等,其中LC-MS和CE-MS联用技术可望成为较有前途的分析方法。:蛋白多肽类药物;药代动力学;分析方法:R972　:A　:1001-0971(2006)01-0063-04　:2005-05-19　:(1975-),,,:,Tel:[phone],E_mail:[e-mail]　＊:,,,,:,Tel:010_66931649　　,、、,SARS、SARS。,,,、,。,:(1),,、;(2),,,,。,,、、、(LC)、(CE)、(GC)、(MS)、(NMR)、-(LC-MS)-(CE-MS)。1　(),()。,。:()。,,,,。,,,、。2　,,,、、。Liu[1](iodogen)[125I](sifu-virtide),,(TCA),,TCA,[125I],。,:,、;;;·63·　20062　331DOI:10.13220/j.cnki.jipr.2006.01.023,,。3　,,。(RIA)、(IRMA)(ELISA)。,ELISA,、、、、、[2]。,;;。,,Hartmann[3]LCRIA(GLP)-2,RIA。4　(RP-HPLC),LC55%,,RP-HPLC,-。Gallwitz[4]GLP-1N1Ⅳ,RP-HPLC,,α--GLP-1,GLP-1NⅣ。Wen[5](LHRH)、,RP-HPLC,LHRH,,。Besser[6]RP-HPLC(somato-stain)-14,-141h,,15h。5　CE、、。,,CE。(CZE),(MECC)。,、。Lacher[7]CE,Cu(Ⅱ),。Zhang[8](CEC),CEC。CE,。1～2;;,,;,,(LIF),,Chen[9](DPDPE),TRITC,CE-LIF1.3×10-9mol·L-1,。6　MS,(Mr)。MS,,。,(FAB)、(ESI)(MALDI),μmol·L-1nmol·L-1Mr,MS-MS、,,、。FAB-MS、。ESI-MS,MSMr,Mr,。García-Garayoa[10]ESI-MS·64·ForeignMedicalSciencesSectionofPharmacy　2006Feb;33(1)(neurotensin)8～13。ESI-MSLCCE,,。MALDI-MS、。mmol·L-1,,,。MALDI-MS,Mr。Wen[11]LHRH,RP-HPLC,UV,MS(MALDI-TOF-MSESI-Q-MS)。7　LC,,,,,。-(LC-MSCE-MS),。7.1　-LC-MS、、Mr,,LC-MS。Sandin[12]RP-HPLC-ESI-MS(SEC)-LC-ESI-MS(BEND),MS-MS,,BEND(1～19)BEND(20～31)。Feng[13]LC-MS-MS,(B201),。2.5～1500μg·L-1,(LOD)0.25μg·L-1,(LOQ)2.5μg·L-1,、、(RSD)。LC-MS,、(SPE),。、、,、、。,。Dai[14,15]SPE-LC-MS,、(OSIM)(DIM),、、,,OSIM-SPE-LC-MS-MS,,300。7.2　-,CEMS、,Mr。Ensing[16]CE-MSY(NPY),,CE-UV。Cao[17]CE-ESI-TOF-MS(hemoglobin)-S,。Li[18]-CE-ESI-MS-CE-MS-MS,CE-MS,,。Preisler[19]CEMALDI-TOF-MS,。,CE-MSLC-MS,。8　8.1　GCGC,。GC-MS,Yi[20]。,20%GC,,。8.2　、NMR,、,NMR。NMR·65·　20062　331、。,LC-NMR,Ⅱ[21]。LC-NMR-MSNMRMS,。9　,10,,,;;,;LC-ESI-MS、LC-MALDI-TOF-MSCE-MS,,Mr,。,,,,,,,。[1]　LiuDS,MengZY,ZhaoGS,etal.Pharmacokineticsof[125I]sifuvir-tideinrats[J].ChinJPharmacolToxicol,2005,19(4):296-299.[2]　LambiaseA,TirassaP,MiceraA,etal.Pharmacokineticsofcon-junctivallyappliednervegrowthfactorintheretinaandopticnerveofadultrats[J].InvestOphthalmolVisSci,2005,46(10):3800-3806.[3]　HartmannB,HarrMB,JeppesenPB,etal.Invivoandinvitrodegradationofglucagon-likepeptide-2inhumans[J].JClinEn-docrinolMetab,2000,85(8):2884-2888.[4]　GallwitzB,RopeterT,Morys-WortmannC,etal.GLP-1-analoguesresistanttodegradationbydipeptidyl-peptidaseⅣinvitro[J].RegulPept,2000,86(1-3):103-111.[5]　WenJY,LedgerR,McLeodBJ,etal.Enzymaticdegradationofluteinizinghormonereleasinghormone(LHRH)bymucosalho-mogenatesfromtheintestineofthecommonbrushtailpossum(Tri-chosurusvulpecula)[J].LifeSci,2002,71(25):3019-3030.[6]　BesserD,MullerB,KleinwachterP,etal.Synthesisandcharacte_rizationofoctapeptidesomatostatinanaloguewithbackbonecycliza-tion:comparisonofdifferentstrategies,biologicalactivitiesandenzy-maticstabilities[J].JPraktChem,2000,342(6):537-545.[7]　LacherNA,GarrisonKE,LunteSM.Separationanddetectionofan-giotensinpeptidesbyCu(Ⅱ)complexationandcapillaryelec-trophoresiswithUVandelectrochemicaldetection[J].Electrophore-sis,2002,23(11):1577-1584.[8]　ZhangK,JiangZ,YaoC,etal.Separationofpeptidesbypressu_rizedcapillaryelectrochromatography[J].JChromatogrA,2003,987(1/2):453-458.[9]　ChenC,JefferyD,JorgensonJW,etal.Sensitiveanalysisof[D-Pen2,5]enkephalininratserumbycapillaryelectrophoresisandlase-rinducedfluorescencedetection[J].JChromatogrBBiomedSciAp-pl,1997,697(1/2):149-162.[10]　García-GarayoaE,Allemann-TannahillL,BläuensteinP,etal.InvitroandInvivoevaluationofnewradiolabeledneurotensin(8-13)analogueswithhighaffinityforNT1receptors[J].NuclMedBiol,2001,28(1):75-84.[11]　WenJ,DaviesN,LedgerR,etal.Isocraticliquidchromatographicassayformonitoringthedegradationofluteinizinghormonereleasinghormonebyextractsfromthegastrointestinaltractofpossums[J].JChromatogrBAnalytTechnolBiomedLifeSci,2002,779(2):221-227.[12]　SandinJ,NylanderI,SilberringJ.Metabolismofβ-endorphininplasmastudiedbyliquidchromatography-electrosprayionizationmassspectrometry[J].RegulPept,1998,73(1):67-72.[13]　FengWY,ChanKK,CoveyJM.ElectrosprayLC-MS/MSquantita-tion,stability,andpreliminarypharmacokineticsofbradykininanta-gonistpolypeptideB201(NSC710295)inthemouse[J].JPharmBiomedAnal,2002,28(3/4):601-612.[14]　DaiSJ,SongHF,DouGF,etal.Quantificationofsifuvirtideinmonkeyplasmabyanon-linesolid-phaseextractionprocedurecom-binedwithliquidchromatography/electrosprayionizationtandemmassspectrometry[J].RapidCommunMassSpectrom,2005,19(10):1273-1282.[15]　DaiSJ,DouGF,QiangXH,etal.Pharmacokineticsofsifuvirtide,anovelanti-HIV-1peptide,inmonkeysanditsinhibitoryconcentra-tioninvitro[J].ActaPharmacolSin,2005,26(10):1274-1280.[16]　EnsingK,deBoerT,SchreuderN,etal.Separationandidentifica-tionofneuropeptideY,twoofitsfragmentsandtheirdegradationproductsusingcapillaryelectrophoresis-massspectrometry[J].JChromatogrBBiomedSciAppl,1999,727(1/2):53-61.[17]　CaoP,MoiniM.Pressure-assistedandpressure-programmedcapil-laryelectrophoresis/electrosprayionizationtimeofflight-massspec-trometryfortheanaly