AG13764 and AG13711 Reverses VEGF-Induced Choroidal Neovascularization in Rat Eye

目的：老年性黄斑变性(AMD)是60岁以上人群致盲的主要原因。在湿态AMD中，针对生长因子信号通路的化合物，如血管内皮生长因子，一直是治疗干预的主要焦点。在先前建立的大鼠CNV模型中，我们在CNV建立后使用了两种受体酪氨酸激酶抑制剂(RTKi)来阻断VEGFR-1、VEGFR-2和PDGFR信号转导。 方法：将AAV-VEGF165注射于出生后15~17天的大鼠视网膜下。6周后，将RTK抑制剂AG013764或AG013711的混悬液注射到腹膜内(ip，每天两次)或玻璃体内(每隔5天)，持续两周。动物处死后制备FITC-葡聚糖全层RPE-脉络膜巩膜。用NeuroLucida对CNV面积进行定量，以测量FITC-葡聚糖整体支架上的超强荧光。如前所述进行组织学和免疫组织化学检查。 结果：免疫组织化学证实对照组和治疗眼均有VEGF的表达，组织学切片显示治疗眼视网膜形态恢复，新生血管减少。在注射AG013764或AG013711的动物中，平均CNV水平比对照组降低了25%~33%，但这一效果没有统计学意义。玻璃体内注射AG013764或AG013711与对照组相比，CNV水平降低了约60%(分别为P&lt；0.005或P&lt；0.05)。结论：这些数据表明，两种RTK抑制剂AG013764或AG013711玻璃体内给药，可显著抑制AAV-VEGF165模型CNV的血管增殖。

Purpose: Age-related macular degeneration (AMD) is the major cause of blindness for people over 60. In the wet form of AMD compounds targeting growth factor signaling pathways such as VEGF have been a major focus for therapeutic interventions. In a previously developed rat model of CNV, we utilized two receptor tyrosine kinase inhibitors (RTKi) to block VEGFR-1, VEGFR-2 and PDGFR signaling following the establishment of CNV. Methods: AAV-VEGF165 was injected into the subretinal space of rats at postnatal days 15-17. Six weeks later, a suspension of RTK inhibitors, AG013764 or AG013711, was injected intraperitoneally (IP, twice daily) or intravitreally (every five days) over a two week period. FITC-dextran whole-mounts of RPE-choroid-sclera were prepared after the animals were sacrificed. CNV area was quantified using Neurolucida to measure the hyperfluorescence on FITC-dextran whole-mounts. Histology and immunohistochemistry were performed as described previously. Results: VEGF expression in control and treated eyes was confirmed by immunohistochemistry and histological sections indicated recovery of retinal morphology and CNV reduction in treated eyes. In the animals injected by IP with AG013764 or AG013711 the mean CNV level was reduced by 25 to 33% compared to control, but this effect did not achieve statistical significance. Intravitreal injections of AG013764 or AG013711 reduced the level of CNV by approximately 60% compared to control (p< 0.005 or p< 0.05, respectively). Conclusions: These data show that two RTK inhibitors, AG013764 or AG013711, delivered intravitreally, significantly reduce blood vessel proliferation in this AAV-VEGF165 model of CNV.