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Updated: Jun 27, 2026
A Simple Bioassay for the Evaluation of Vascular Endothelial Growth Factors
Published on: March 15, 2016
Identification of a novel vascular endothelial growth factor receptor 2 inhibitor and its effect for choroidal
Hidenori Takahashi1, Yasuhiro Tamaki, Nobuya Ishii
1Department of Ophthalmology, University of Tokyo School of Medicine, Bunkyo-ku, Tokyo, Japan.
Purpose:
To select a novel orally administered VEGFR-2 (KDR/flk-1) specific tyrosine kinase inhibitor in a murine model of choroidal neovascularization (CNV).
Methods:
From a compound library, potent VEGFR2 inhibitors were selected by VEGF-induced phosphorylation of VEGFR-2 and RAF kinases and the proliferation analysis by HUVEC cultures and in vitro tube formation assay. CNV was induced in C57/BL6 mice using diode laser photocoagulation. The antiangiogenic effect of selected compounds was assessed by angiographic examination, in which extent of fluorescein leakage was scored and histological analysis, allowing for measurement of CNV membrane under light microscope. In addition, C57/BL6 mice were treated with daily oral administration of selected compounds for 14 days and body weights were measured.
Results:
Six compounds that potently inhibited VEGFR-2 were selected for further investigation. Selected compounds-treated conditions showed a dose-dependent inhibition of phosphorylation of VEGFR-2 tyrosine kinase with an IC50 of 0.0022 to 0.098 microm. Selected compounds did not inhibit the HCT116 proliferation but did demonstrate a strong inhibition effect for VEGFR-2 dependant HUVEC (IC50=0.0018 to 0.058 microm). Selected compounds treatment also resulted in a dose-dependent attenuation of in vitro tube formation. In the murine CNV model, #0451 is the most effective compound. The intensity of fluorescein leakage was significantly lower in doses of 12.5, 25, 50, and 100 mg/kg #0451-treated eyes compared to controls. Histologically, CNV membrane volumes were significantly reduced in #0451-treated eyes in a dose-dependent manner. At therapeutic doses of 100 mg/kg or less, there was no significant weight loss between the treated and untreated groups.
Conclusion:
Oral administration of #0451, a novel VEGFR-2 (KDR/flk-1)-specific tyrosine kinase inhibitor, demonstrates anti-angiogenic effects in our murine model of CNV. #0451 may be useful to treat the choroidal neovascularization associated with AMD.
Insights
A novel oral tyrosine kinase inhibitor, #0451, effectively reduced choroidal neovascularization (CNV) in mice. This VEGFR-2 specific compound demonstrated anti-angiogenic effects without significant weight loss, suggesting potential for treating AMD-related CNV.
Area of Science:
- Ophthalmology
- Pharmacology
- Angiogenesis Research
Background:
- Choroidal neovascularization (CNV) is a key factor in vision loss associated with age-related macular degeneration (AMD).
- Targeting vascular endothelial growth factor receptor 2 (VEGFR-2) is a promising strategy for anti-angiogenic therapy.
Purpose of the Study:
- To identify and evaluate a novel, orally administered VEGFR-2 specific tyrosine kinase inhibitor for treating CNV.
- To assess the efficacy and safety of selected compounds in a murine model of CNV.
Main Methods:
- Compounds were screened for VEGFR-2 inhibition and anti-proliferative effects on HUVECs.
- CNV was induced in mice via laser photocoagulation.
- Anti-angiogenic effects were evaluated using fluorescein angiography and histological analysis; body weight was monitored.
Main Results:
- Six potent VEGFR-2 inhibitors were identified, with significant dose-dependent inhibition of VEGFR-2 phosphorylation and HUVEC proliferation.
- Compound #0451 showed the most potent anti-angiogenic activity in the murine CNV model, significantly reducing fluorescein leakage and CNV volume.
- #0451 was well-tolerated at therapeutic doses, with no significant weight loss observed.
Conclusions:
- Oral administration of #0451, a novel VEGFR-2 specific tyrosine kinase inhibitor, exhibits significant anti-angiogenic effects in a murine CNV model.
- #0451 demonstrates therapeutic potential for treating choroidal neovascularization associated with AMD.
