The use of one-bead one-compound combinatorial library technology to discover high-affinity αvβ3 integrin and cancer targeting arginine-glycine-aspartic acid ligands with a built-in handle.
The αvβ3 integrin, expressed on the surface of various normal and cancer cells, is involved in numerous physiologic processes such as angiogenesis, apoptosis, and bone resorption. Because this integrin plays a key role in angiogenesis and metastasis of human tumors, αvβ3 integrin ligands are of great interest to advances in targeted therapy and cancer imaging. In this report, one-bead one-compound (OBOC) combinatorial libraries containing the arginine-glycine-aspartic acid (RGD) motif were designed and screened against K562 myeloid leukemia cells that had been transfected with the human αvβ3 integrin gene. Cyclic peptide LXW7 was identified as a leading ligand with a built-in handle that binds specifically to αvβ3 and showed comparable binding affinity (IC(50) = 0.68 ± 0.08 μmol/L) to some of the well-known RGD "head-to-tail" cyclic pentapeptide ligands reported in the literature. The biotinylated form of LXW7 ligand showed similar binding strength as LXW7 against αvβ3 integrin, whereas biotinylated RGD cyclopentapeptide ligands revealed a 2- to 8-fold weaker binding affinity than their free forms. LXW7 was able to bind to both U-87MG glioblastoma and A375M melanoma cell lines, both of which express high levels of αvβ3 integrin. In vivo and ex vivo optical imaging studies with the biotinylated ligand/streptavidin-Cy5.5 complex in nude mice bearing U-87MG or A375M xenografts revealed preferential uptake of biotinylated LXW7 in tumor. When compared with biotinylated RGD cyclopentapeptide ligands, biotinylated LXW7 showed higher tumor uptake but lower liver uptake.
Xiao
Wenwu
W
Department of Biochemistry and Molecular Medicine, University of California Davis, Sacramento, California 95817, USA.
Wang
Yan
Y
Lau
Edmond Y
EY
Luo
Juntao
J
Yao
Nianhuan
N
Shi
Changying
C
Meza
Leah
L
Tseng
Harry
H
Maeda
Yoshiko
Y
Kumaresan
Pappanaicken
P
Liu
Ruiwu
R
Lightstone
Felice C
FC
Takada
Yoshikazu
Y
Lam
Kit S
KS
D000818:Animals; D020650:Combinatorial Chemistry Techniques; D005434:Flow Cytometry; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000378:metabolism*; D020014:K562 Cells; D008024:Ligands; D051379:Mice; D008819:Mice, Nude; D009369:Neoplasms / Q000378:metabolism*; D009842:Oligopeptides / Q000378:metabolism*; D014183:Transplantation, Heterologous
D039302:Integrin alphaVbeta3; D008024:Ligands; D009842:Oligopeptides