Oh Eun Ju, Park Kitae, Choi Jun-Sub, Joo Choun-Ki, Hahn Sei Kwang
Department of Materials Science and Engineering, Pohang University of Science and Technology (POSTECH), Pohang, Kyungbuk, South Korea.
Biomaterials. 2009 Oct;30(30):6026-34. doi: 10.1016/j.biomaterials.2009.07.024. Epub 2009 Jul 31.
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.
据报道，GNQWFI抗Flt1肽可抑制血管内皮生长因子受体1（VEGFR1）介导的内皮细胞迁移和管腔形成。在本研究中，成功开发了一种合成抗Flt1肽-透明质酸（HA）偶联物的方案，用于治疗角膜新生血管。使用HA的四丁铵盐（HA-TBA），通过HA的羧基与GGNQWFI的N端胺基之间形成酰胺键，可使水不溶性抗Flt1肽与HA在二甲基亚砜（DMSO）中偶联。通过（1）H NMR和荧光分析证实了抗Flt1肽-HA偶联物的形成。通过改变偶联反应中肽的进料量，可将抗Flt1肽-HA偶联物中每个单链HA上接枝的肽分子平均数控制在3至30个之间。根据体外生物活性测试，抗Flt1肽-HA偶联物对包被在孔中的Flt1-Fc与VEGF(165)的结合表现出显著的抑制作用。此外，通过对SD大鼠硝酸银烧灼角膜中角膜新生血管的抑制作用，证实了抗Flt1肽-HA偶联物的体内生物活性。用抗Flt1肽-HA偶联物处理后，VEGF受体2的表达也降低了。水溶性抗Flt1肽-HA偶联物被认为有潜力开发成为治疗角膜新生血管的抗血管生成疗法。
