Synthesis, characterization, and preliminary assessment of anti-Flt1 peptide-hyaluronate conjugate for the treatment of corneal neovascularization.
Anti-Flt1 peptide of GNQWFI has been reported to inhibit vascular endothelial growth factor receptor 1 (VEGFR1) - mediated endothelial cell migration and tube formation. In this work, a protocol to synthesize anti-Flt1 peptide-hyaluronate (HA) conjugate was successfully developed for the treatment of corneal neovascularization. Using tetrabutyl ammonium salt of HA (HA-TBA), water-insoluble anti-Flt1 peptide could be conjugated with HA in dimethyl sulfoxide (DMSO) by the amide bond formation between carboxyl groups of HA and N-terminal amine groups of GGNQWFI. The formation of anti-Flt1 peptide-HA conjugate was confirmed by (1)H NMR and fluorometric analyses. The average number of grafted peptide molecules in anti-Flt1 peptide-HA conjugates could be controlled from 3 to 30 per single HA chain by changing the feeding amount of peptide for the conjugation reaction. According to in vitro biological activity tests, anti-Flt1 peptide-HA conjugate exhibited a significant inhibition effect on the binding of Flt1-Fc to VEGF(165) coated on the well. Furthermore, in vivo biological activity of anti-Flt1 peptide-HA conjugate was confirmed from the inhibitory effect on corneal neovascularization in silver nitrate cauterized corneas of SD rats. The VEGF receptor 2 expression was also reduced after treatment with anti-Flt1 peptide-HA conjugate. The water-soluble anti-Flt1 peptide-HA conjugate was thought to have a potential to be developed as anti-angiogenic therapeutics for the treatment of corneal neovascularization.
Oh
Eun Ju
EJ
Department of Materials Science and Engineering, Pohang University of Science and Technology (POSTECH), Pohang, Kyungbuk, South Korea.
Park
Kitae
K
Choi
Jun-Sub
JS
Joo
Choun-Ki
CK
Hahn
Sei Kwang
SK
D020533:Angiogenesis Inhibitors / Q000494:pharmacology; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D002465:Cell Movement; D016510:Corneal Neovascularization / Q000378:metabolism*; D015195:Drug Design; D042783:Endothelial Cells / Q000166:cytology; D005470:Fluorometry / Q000379:methods; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000737:chemistry / Q000276:immunology*
D020533:Angiogenesis Inhibitors; D001672:Biocompatible Materials; D010455:Peptides; D000644:Quaternary Ammonium Compounds; D006820:Hyaluronic Acid; C501162:FLT1 protein, human; D040281:Vascular Endothelial Growth Factor Receptor-1