目的合成RGD(精氨酸-甘氨酸-天门冬氨酸)肽类似物-AoGDw(ω-氨基辛酸-甘氨酸-天门冬氨酸-色氨酸),研究其结构对血小板聚集功能的影响。方法采用固相法合成AoGDW,经Sephadex G-10纯化,RP-HPLC(反相高效液相色谱法)、质谱和氨基酸组成分析对其进行鉴定,并采用比浊法测定其抗血小板聚集的活性及稳定性。结果获得了纯度为98.3%的目的肽、质谱和氨基酸组成分析结果均与理论值一致。AoGDW及对照组抑制血小板聚集的IC50分别为(4.7l±2.51)和(61.82±8.08)μmol·L-1在血浆中孵育3 h,仍保持100%的活性。结论AoGDW具有更强的抗血小板聚集的活性及稳定性,说明与RGD序列紧邻的氨基酸残基对其功能有重要的影响。
Abstract
OBJECTIVE To synthesize RGD(arginine-glycine-aspartic acid-tryptophan)peptide analog-AoGDW(ω-amino- octanoic acid-glycine-aspartic acid-tryptophan)and investigate its effect on inhibiting platelet aggregation.METHODS AoGDW was synthesized by solid phase method and purified on Sephadex G-10.The synthesized peptide was identified by RP-HPLC(reversed- phase high performance liquid chromatography).The structure of peptide was confirmed by MS(mass spectrograph)and amino acid a- nalysis.The activity and stability of antiplatelet aggregation of AoGDW were studied using aggregometer and expressed as percentage change in light transmittance.RESULTS The purity of purified peptide was 98.3%.Peptide structure verified through MS and ami- no acid analysis was consistent with theory.IC50 values of inhibiting platelet aggregation of AoGDW and control group were(4.71±2.51)and(61.82±8.08)μmol·L-1.AoGDW was achieved with improved plasma stability(100% activity after 3 h).CONCLU- SION AoGDW was highly active,highly steady inhibitor of platelet aggregation,which suggests the side chains and backbone ele- ments of residues flanking the RGD sequence may play very important effects on its function.
关键词
精氨酸-甘氨酸-天门冬氨酸 /
固相法 /
血小板
{{custom_keyword}} /
Key words
RGD /
solid-phase method /
platelet /
activities
{{custom_keyword}} /
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] TAKAGI J. Structural basis for ligand recognition by RGD (Arg-Gly-Asp)-dependent integrins[J] . Biochem Soc Trans, 2004, 32：403-406.
[2] MEYER A, AUERNHEIMER J, MODLINGER A, et al. Targeting RGD recognizing integrins: drug development, biomaterial research, tumor imaging and targeting[J] . Curr Pharm Des, 2006, 12(22):2723-2747.
[3] DUNEHOO A L, ANDERSON M, MAJUMDAR S, et al. Cell adhesion molecules for targeted drug delivery[J] . J Pharm Sci, 2006, 95(9):1856-1872.
[4] FOSTER M R, HORNBY E J, BROWN S, et al. Improved potency and specificity of Arg-Gly-Asp (RGD) containing peptides as fibrinogen receptor blocking drugs[J] . Thromb Res, 1993, 72(3):231-245.
[5] CHENG S, CRAIG W S, MULLEN D, et al. Design and synthesis of novel cyclic RGD-containing peptides as highly potent and selective integrin alpha Iib beta 3 antagonists[J] . J Med Chem, 1994, 37(1):1-8.
[6] RUOLAHTI E，PIESCHBACHER M D.New perspectives in cell adhesion: RGD and integrins[J] . Science, 1987, 238(4826):491-497.
[7] TOMIYAMA Y, BROJER E, RUGGERI Z M, et al. A molecular model of RGD ligands. Antibody D gene segments that direct specificity for the integrin alpha Iib beta 3[J] . J Biol Chem, 1992, 267(25):18085-18092.
[8] FOK K F, PANZER-KNODLE S G, NICHOLSO N S, et al. Aminopeptidase resistant Arg-Gly-Asp analogs are stable in plasma and inhibit platelet aggregation[J] . Int J Pept Protein Res, 1991, 38(2):124-130.
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
