Inhibition of inflammatory corneal angiogenesis by TNP-470

PURPOSE. To determine the efficacy of the angiogenic inhibitor TNP-470 on i nflammatory corneal neovascularization. Topical and systemic delivery of th e drug were investigated in a murine model as well as inhibition of endothe lial cell proliferation in vitro and in vivo. METHODS. The effect of TNP-470 on VEGF- and bFGF-stimulated bovine capillar y endothelial (BCE) cell proliferation was evaluated in vitro. Corneal neov ascularization was induced in vivo by mechanical debridement of the corneal and limbal epithelium with 0.15 M NaOH on C57BL6 mice. TNP-470 was adminis tered systemically at 30 mg/kg body weight (BW) every other day or topicall y three times daily in a concentration of 5 ng/ml dissolved in methylcellul ose. Vessel length was investigated on day 7. VEGF protein content in murin e corneas was analyzed by ELISA on days 2, 4, and 7 of treatment. A modifie d bromouridine (BrdU) ELISA was used to quantify endothelial cell prolifera tion. RESULTS. TNP-470 exerted a dose-dependent inhibition of bFGF-and VEGF-induc ed endothelial cell proliferation in vitro. Both systemic and topical appli cation of TNP-470 led to a significant reduction of inflammatory corneal ne ovascularization (P < I X 10(-5)). BrdU labeling showed that TNP-470 inhibi ted endothelial cell proliferation. VEGF protein levels were reduced by sys temic TNP-470 treatment. CONCLUSIONS. These results suggest that TNP-470 reduces inflammatory cornea l angiogenesis by directly inhibiting endothelial cell proliferation. Topic al and systemic treatment with TNP-470 reduces VEGF levels that are respons ible for vessel growth during the neovascularization process.