Use of heparinase to decrease inflammatory responses

Abstract

Heparinase enzymes can be used as a medical treatment to reduce localized inflammatory responses. Treatment of activated endothelium with heparinase inhibits leukocyte rolling, adhesion and extravasation. Most of the heparin and heparan sulfate on endothelial cell surfaces and in basement membranes is degraded by exposure to heparinase. In addition, immobilized chemokines, which are attached to heparin/heparan sulfate on activated endothelium are solubilized by heparinase digestion. Heparinase can be infused into the vascular system to inhibit accumulation of leukocytes in inflamed tissue and decrease damage resulting from localized inflammations. Targeting of heparinase to activated endothelium can be accomplished through localized administration and/or use of genetically engineered heparinase containing endothelium ligand-binding domains.

What is claimed is:  
     
         1 . A method to decrease localized inflammatory responses in a tissue of a patient comprising administering to said patient heparinase enzyme in an effective amount sufficient to decrease said localized inflammatory response.  
     
     
         2 . The method of    claim 1   , wherein said administration of said heparinase enzyme removes and digests heparin and heparan sulfate from endothelial cell surfaces and extracellular matrices of said tissue.  
     
     
         3 . The method of    claim 1   , wherein said administration of said heparinase enzyme decreases the accumulation of leukocytes in tissue adjacent to endothelial cell surfaces and extracellular matrices.  
     
     
         4 . The method of    claim 1   , wherein said administration of said heparinase enzyme inhibits leukocyte extravasation by releasing immobilized chemokines and destroying chemokines immobilized to endothelium.  
     
     
         5 . The method of    claim 1   , wherein said administration of said heparinase enzyme inhibits leukocyte rolling on endothelium.  
     
     
         6 . The method of    claim 1   , wherein said heparinase enzyme is overexpressed from a recombinant nucleotide sequence, in  Flavobacterium heparinum.    
     
     
         7 . The method of    claim 1   , wherein said heparinase enzyme is expressed from a recombinant nucleotide sequence in an organism in which it does not naturally occur.  
     
     
         8 . A method to decrease a localized inflammatory response in a tissue of a patient comprising administering to said patient a fusion protein comprising a ligand which binds to activated endothelial cells and a heparinase enzyme in an amount sufficient to decrease said localized inflammatory response in said tissue.  
     
     
         9 . The method of    claim 8   , wherein said fusion protein is made by genetic engineering techniques.  
     
     
         10 . A pharmaceutical composition comprising a heparinase enzyme together with a pharmaceutically or a veterinarilly acceptable carrier.  
     
     
         11 . A pharmaceutical composition comprising fusion molecule comprising a ligand which binds to activated endothelium and a heparinase enzyme.  
     
     
         12 . The pharmaceutical composition of    claim 11   , wherein said ligand binding domain is selected from the group consisting of cytokines, antibodies, integrins, and selecting.  
     
     
         13 . The pharmaceutical composition of    claim 11   , wherein said ligand binding domain are fragments of said cytokines, antibodies, integrins, and selecting.  
     
     
         14 . The pharmaceutical composition of    claim 11   , wherein said fragments are selected from the group consisting of cytokine receptor binding domains, Fab fragments, antibody variable regions, integrin I-domains, and selectin domains.  
     
     
         15 . The pharmaceutical composition of either claims  10  or  11 , wherein said carrier is selected from the group consisting of liposomes, lipospheres, proteosomes, microspheres, microcapsules, and biodegradable polymeric matrices.  
     
     
         16 . The use of a heparinase enzyme in the preparation of a medicant for treatment of decreasing localized inflammatory responses in a patient's tissue.  
     
     
         17 . The use of a heparinase enzyme in the preparation of a medicant comprising a fusion protein comprising a ligand which binds to activated endothelial cells and a heparinase enzyme for the treatment of decreasing localized inflammatory responses in a patient's tissue.