We plan to make a fluorescent marker for neovascularization. Specifically, we want to stain new blood vessels with a fluorescent analogue of fluorescein. This would be useful in assessing diabetic retinal changes and also in assessing limited subsets of age related macular degeneration. We have found a specific marker for neovascularization, being UPA. We will then find and develop a fluorescein compound, specifically to label the UPA present on new vessels. Two classes have been proposed with some apparent preliminary success:(1) Amiloride plus fluorescein, since Amiloride has UPA specificity and inhibitory properties, and(2) amino fluorescein, which has less avid binding of UPA, but has some specificity. We will test these compounds four ways:(1) protease inhibition for UPA, TPA, and a variety of other proteases,(2) the dissociative constant to find the usefulness of the drug,(3) albumin binding to check fluorescence as well as to check any change in emission and absorption properties of the new compound, and(4) test its binding to UPA in the wounded endothelial cell monolayer culture system.Awardee's statement of the potential commercial applications of the research:The major product of this research is a targeted fluorescein analog which would provide more selective visualization of neovascularization by fluorescein angiography. Such a compound would have potential as a replacement for fluorescein in the most widely used retinal diagnostic procedure and as a research reagent in the basic studies of neovascularization.National Eye Institute (NEI)
