TARGETED FLUORESCEIN DYES FOR ANGIOGRAPHY

WE PLAN TO MAKE A FLUORESCENT MARKER FOR NEOVASCULARIZATION. SEPCIFICALLY, WE WANT TO STAIN NEW BLOOD VESSELS WITH FLUORESCENT ANALOGUE OF FLUORESCEIN. THIS WOULD BE USEFUL IN ASSESSING DIABETIC RETINAL CHANGES AND ALSO IN LIMITED SUBSETS OF AGE RELATED MACULAR DEGERNATION. 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 (AMILORIDE HAS UPA SPECIFICITY AND INHIBITORY PROPERTIES) AND (2) AMINO FLUORESCEIN (HAS LESS AVID BINDING OF UPA BUT SOME SPECIFICITY). WE WILL TEST THESE COMPOUNDS FOUR WAYS: (1) PROTEASE INHIBITION FOR UPA, TPA, A VARIETY OF OTHER PROTEASES, (2) THE DISSOCIATIVE CONSTANT TO FIND USEFULNESS OF THE DRUG, (3) ALBUMIN BINDING TO CHECK FLUORESCENT 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.