Preparation and functional evaluation of RGD-modified proteins as αvβ3 integrin directed therapeutics

被引:144
作者
Kok, RJ [1 ]
Schraa, AJ [1 ]
Bos, EJ [1 ]
Moorlag, HE [1 ]
Asgeirsdóttir, SA [1 ]
Everts, M [1 ]
Meijer, DKF [1 ]
Molema, G [1 ]
机构
[1] Univ Groningen, IDE, Dept Pharmacokinet & Drug Delivery, Univ Ctr Pharm, NL-9713 AV Groningen, Netherlands
关键词
D O I
10.1021/bc015561+
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Tumor blood vessels can be selectively targeted by RGD-peptides that bind to alpha(v)beta(3) integrin on angiogenic endothelial cells. By inhibiting the binding of these integrins to its natural ligands, RGD-peptides can serve as antiangiogenic therapeutics. We have prepared multivalent derivatives of the cyclic RGD-peptide c(RGDfK) by covalent attachment of the peptide to side chain amino groups of a protein. These RGDpep-protein conjugates inhibited alpha(v)beta(3)-mediated endothelial cell adhesion in vitro, while conjugates prepared with a control RAD-peptide showed no activity. Radiobinding and displacement studies with endothelial cells demonstrated an increased affinity of the RGDpep-protein conjugates compared to the free peptide, with IC50 values ranging from 23 to 0.6 nM, depending on the amount of coupled RGDpep per protein. Compared to the parental RGD-peptide and the related RGD-peptide ligand c(RGDfV), the RGDpep-protein conjugates showed a considerable increase in affinity (IC50 parent RGDpep: 818 nM; IC50 c(RGDfV): 158 nM). We conclude that the conjugation of RGD-peptides to a protein, resulting in products that can bind multivalently, is a powerful approach to increase the affinity of peptide ligands for alpha(v)beta(3)/alpha(v)beta(5) integrins.
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页码:128 / 135
页数:8
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