2′-Fluoropyrimidine RNA-based Aptamers to the 165-Amino Acid Form of Vascular Endothelial Growth Factor (VEGF165)

• Published in: The Journal of biological chemistry Vol. 273; no. 32; p. 20556
• Main Authors: Judy Ruckman, Louis S. Green, Jim Beeson, Sheela Waugh, Wendy L. Gillette, Dwight D. Henninger, Lena Claesson-Welsh, Nebojsa Janjic
• Format: Journal Article
• Language: English
• Published: American Society for Biochemistry and Molecular Biology 07.08.1998
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.273.32.20556

Vascular endothelial growth factor (VEGF) has been implicated in the pathological induction of new blood vessel growth in a variety of proliferative disorders. Using the SELEX process ( s ystematic e volution of l igands by ex ponential enrichment), we have isolated 2′-F-pyrimidine RNA oligonucleotide ligands (aptamers) to human VEGF 165 . Representative aptamers from three distinct sequence families were truncated to the minimal sequence capable of high affinity binding to VEGF (23–29 nucleotides) and were further modified by replacement of 2′- O -methyl for 2′-OH at all ribopurine positions where the substitution was tolerated. Equilibrium dissociation constants for the interaction of VEGF with the truncated, 2′- O -methyl-modified aptamers range between 49 and 130 p m . These aptamers bind equally well to murine VEGF 164 , do not bind to VEGF 121 or the smaller isoform of placenta growth factor (PlGF 129 ), and show reduced, but significant affinity for the VEGF 165 /PlGF 129 heterodimer. Cysteine 137 in the exon 7-encoded domain of VEGF 165 forms a photo-inducible cross-link to a single uridine residue in each of the three aptamers. The aptamers potently inhibit the binding of VEGF to the human VEGF receptors, KDR and Flt-1, expressed by transfected porcine aortic endothelial cells. Furthermore, one of the aptamers is able to significantly reduce intradermal VEGF-induced vascular permeability in vivo .