Preparation and functional evaluation of RGD-modified proteins as αvβ3 integrin directed therapeutics
Preparation and functional evaluation of RGD-modified proteins as αvβ3 integrin directed therapeutics
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DOI：
10.1021/bc015561
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发表时间：
2002-01-01
影响因子：
4.7
通讯作者：
Molema, G
中科院分区：
文献类型：
--
作者：
Kok, RJ;Schraa, AJ;Molema, G
肿瘤血管可以被rgd -肽选择性靶向，rgd -肽与血管生成内皮细胞上的α (v) β(3)整合素结合。通过抑制这些整合素与其天然配体的结合，rgd -肽可以作为抗血管生成治疗药物。我们制备了环rgd -肽c（RGDfK）的多价衍生物，通过多肽共价连接到蛋白质的侧链氨基。这些rgdpep -蛋白偶联物在体外抑制α (v) β(3)介导的内皮细胞粘附，而用对照rad -肽制备的偶联物则没有活性。内皮细胞的放射结合和位移研究表明，与游离肽相比，RGDpep-蛋白偶联物的亲和力增加，IC50值从23到0.6 nM不等，这取决于每个蛋白偶联RGDpep的量。与亲本rgd -肽和相关rgd -肽配体c（RGDfV）相比，RGDpep蛋白偶联物的亲和力显著提高（IC50亲本RGDpep: 818 nM； IC50 c(RGDfV): 158 nM）。我们得出结论，rgd -多肽与蛋白质结合，产生可多价结合的产物，是一种增强肽配体对α (v) β (3)/ α (v) β(5)整合素亲和力的有效方法。
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.