title: Synthesis, characterization, and preliminary assessment of anti-Flt1 peptide-hyaluronate conjugate for the treatment of corneal neovascularization.
abstract: 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.
authors: Department of Materials Science and Engineering, Pohang University of Science and Technology (POSTECH), Pohang, Kyungbuk, South Korea.
authors: Eun Ju
authors: EJ
authors: Oh
authors: Kitae
authors: K
authors: Park
authors: Jun-Sub
authors: JS
authors: Choi
authors: Choun-Ki
authors: CK
authors: Joo
authors: Sei Kwang
authors: SK
authors: Hahn
mesh_terms: 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*
chemical_list: 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