TARGETING OF UROKINASE TO FIBRIN CLOTS

THE LONG-TERM OBJECTIVE OF THIS PROJECT IS THE DEVELOPMENT AND CHARACTERIZATION OF EFFECTIVE CLOT-DISSOLVING THERAPY AND, IN PARTICULAR, THE STUDY OF A CONJUGATE BETWEEN HUMAN UROKINASE AND A MONOCLONAL ANTIBODY SPECIFIC FOR HUMAN FIBRIN. THIS COMPLEX WOULD ALLOW FOR ENHANCED PLASMINOGEN ACTIVATION AT THE SITES OF FIBRIN CLOTS WHILE REDUCING SYSTEMIC ACTIVATION OF PLASMINOGEN AND THE ASSOCIATED DELETERIOUS EFFECTS. THE INITIAL GOAL OF THESE STUDIES WILL BE EXAMINE THE IMMUNOCHEMISTRY OF HUMAN FIBRIN AND FIBRINOGEN, USING POLYCLONAL RABBIT ANTISERUM TO HUMAN FIBRIN AND EXTENSIVE ASORPTION WITH HUMAN FIBRINOGEN, WITH THE OBJECTIVE OF FINDING EPITOPES UNIQUE TO FIBRIN. SECONDLY, THE PLASMINOGEN ACTIVATING ACTIVITY OF HUMAN UROKINASE THAT HAS BEEN COUPLED TO POLYCLONAL RABBIT ANTI- HUMAN FIBRINOGEN WILL BE INVESTIGATED. STANDARD COUPLING PROCEDURES WILL BE EMPLOYED, AND THE ANTIGENIC SPECIFICITY, ENZYMATIC ACTIVITY, AND SUSCEPTIBILITY TO PROTEASE INHIBITORS OF THE RESULTING COMPLEXES WILL BE DETERMINED. INADDITION TO POTENTIAL DEVELOPMENT OF AN IMPROVED THROMBOLYTIC AGENT FOR CLINICAL USE, THESE STUDIES MAY LEAD TO THE DEVELOPMENT OF A MORE GENERAL APPROACH FOR TARGETING SPECIFIC ENZYMES TO PATHOLOGICAL SITES.