Department of Ophthalmology, College of Medicine, College of Pharmacy, The Ohio State University, Columbus, OH 43210, USA.
Exp Eye Res. 2010 Dec;91(6):837-43. doi: 10.1016/j.exer.2010.10.013. Epub 2010 Nov 3.
The aim of this study was to test the selectivity, in-vivo effectiveness, and potential mechanism of action of a linomide analogue (N-phenyl-1,2-dihydro-4-hydroxyl-2-oxo-quinoline-3-carboxamide, Lin05) for inhibition of choroidal neovascularization. The selectivity of Lin05 was tested in cell proliferation assays with human umbilical vein endothelial cells (HUVEC) and a retinal pigmented epithelial cell line(ARPE-19). In-vivo anti-angiogenic effect of Lin05 was investigated utilizing an experimental laser-induced choroidal neovascularization (ECNV) model in adult Brown Norway rats. Western blot and/or reverse transcriptase-PCR was used to test the effect of Lin05 on potential targets. Our results indicate that Lin05 is at least an 8-fold more selective inhibitor of endothelial cell proliferation compared to RPE cells. Systemic administration of Lin05 in an ECNV model was associated with a significant decrease in both vascular leakage on fluorescein angiography and lesion size by histopathology (p = 0.02). No systemic toxicity was detected for Lin05 in major organs such as the liver, lung and kidneys. Lin05 did not inhibit VEGF-induced VEGFR2 (KDR) phosphorylation in HUVEC nor was associated with decreased VEGF gene expression. Also it did not inhibit insulin-like growth factor (IGF-1) and Epidermal Growth Factor (EGF) induced activation of p42/p44 MAPK activation. It inhibited both PDGF- and bFGF-induced p42/p44 MAPK phosphorylation. However, the effect on PDGF was variable in different HUVEC cells. In conclusion, Lin05 is a potential anti-angiogenic agent for the treatment of eye diseases associated with pathological neovascularization. The anti-angiogenic effect of Lin05 is likely through inhibition of bFGF but not through inhibition of the VEGF/KDR pathway.
本研究旨在测试一种来氟米特类似物（N-苯基-1,2-二氢-4-羟基-2-氧代-喹啉-3-甲酰胺，Lin05）抑制脉络膜新生血管形成的选择性、体内有效性和潜在作用机制。Lin05 的选择性在人脐静脉内皮细胞（HUVEC）和视网膜色素上皮细胞系（ARPE-19）的细胞增殖测定中进行了测试。利用成年棕色挪威大鼠实验性激光诱导脉络膜新生血管（ECNV）模型，研究了 Lin05 的体内抗血管生成作用。Western blot 和/或逆转录-PCR 用于测试 Lin05 对潜在靶点的影响。我们的结果表明，Lin05 对内皮细胞增殖的选择性至少是 RPE 细胞的 8 倍。在 ECNV 模型中，Lin05 的全身给药与荧光素血管造影中的血管渗漏和组织病理学中的病变大小均显著减少（p = 0.02）相关。在肝脏、肺和肾脏等主要器官中未检测到 Lin05 的全身毒性。Lin05 不会抑制 HUVEC 中 VEGF 诱导的 VEGFR2（KDR）磷酸化，也不会导致 VEGF 基因表达减少。它也不会抑制胰岛素样生长因子（IGF-1）和表皮生长因子（EGF）诱导的 p42/p44 MAPK 激活。它抑制 PDGF 和 bFGF 诱导的 p42/p44 MAPK 磷酸化。然而，其对不同 HUVEC 细胞中 PDGF 的作用是可变的。总之，Lin05 是一种有潜力的抗血管生成药物，可用于治疗与病理性新生血管形成相关的眼部疾病。Lin05 的抗血管生成作用可能是通过抑制 bFGF 而不是通过抑制 VEGF/KDR 途径实现的。
