Composition and method for enhancing fibrinolysis

Abstract

The present invention relates to a novel alpha-2-antiplasmin-binding molecules and treatment for pulmonary embolism, myocardial infarction, thrombosis or stroke in a patient which comprises administering an alpha-2-antiplasmin-binding molecule capable of preventing inhibition of plasmin by endogenous alpha-2-antiplasmin. The invention also relates to a treatment for pulmonary embolism, myocardial infarction, thrombosis or stroke in a patient comprising coadministrating an alpha-2-antiplasmin-binding molecule of the invention together with a thrombolytic agent.

What is claimed is:  
     
         1 . An immunologic molecule wherein said immunologic molecule is capable of binding to both (1) human and nonhuman circulating α2-antiplasmins and (2) human and nonhuman fibrin crosslinked α2-antiplasmins.  
     
     
         2 . The immunologic molecule of  claim 1 , wherein said immunologic molecule is a chimeric antibody.  
     
     
         3 . The immunologic molecule of  claim 1 , wherein said immunologic molecule is a humanized antibody.  
     
     
         4 . The immunologic molecule of  claim 1 , wherein said immunologic molecule is an antibody fragment.  
     
     
         5 . The immunologic molecule of  claim 1 , wherein said immunologic molecule is a monoclonal antibody.  
     
     
         6 . The immunologic molecule of  claim 1 , wherein said immunologic molecule comprises amino acids 1 to 107 of SEQ ID NO: 9 and amino acids 1 to 119 of SEQ ID NO: 15.  
     
     
         7 . The immunologic molecule of  claim 1 , wherein said immunologic molecule comprises amino acids 1 to 107 of SEQ ID NO: 5 and amino acids 1 to 119 of SEQ ID NO: 11.  
     
     
         8 . The immunologic molecule of  claim 1 , wherein said immunologic molecule comprises amino acids 1 to 107 of SEQ ID NO: 7 and amino acids 1 to 119of SEQ ID NO: 13.  
     
     
         9 . The immunologic molecule of  claim 1 , selected from the group consisting of: 
 (a) an immunologic molecule, wherein the CDR1 region of the light chain of said immunologic molecule comprises amino acids 26 to 32 of SEQ ID NO: 75;    (b) an immunologic molecule, wherein the CDR2 region of the light chain of said immunologic molecule comprises amino acids 50 to 52 of SEQ ID NO: 75;    (c) an immunologic molecule, wherein the CDR3 region of the light chain of said immunologic molecule comprises amino acids 91 to 96 of SEQ ID NO: 75;    (d) an immunologic molecule, wherein the CDR1 region of the heavy chain of said immunologic molecule comprises amino acids 26 to 32 of SEQ ID NO: 79;    (e) an immunologic molecule, wherein the CDR2 region of the heavy chain of said immunologic molecule comprises amino acids 53 to 56 of SEQ ID NO: 79; and    (f) an immunologic molecule, wherein the CDR3 region of the heavy chain of said immunologic molecule comprises amino acids 100 to 107 of SEQ ID NO: 79.    
     
     
         10 . The immunologic molecule of  claim 1 , selected from the group consisting of: 
 (a) an immunologic molecule, wherein the CDR1 region of the light chain of said immunologic molecule comprises amino acids 26 to 32 of SEQ ID NO: 76;    (b) an immunologic molecule, wherein the CDR2 region of the light chain of said immunologic molecule comprises amino acids 50 to 52 of SEQ ID NO: 76;    (c) an immunologic molecule, wherein the CDR3 region of the light chain of said immunologic molecule comprises amino acids 91 to 96 of SEQ ID NO: 76;    (d) an immunologic molecule, wherein the CDR1 region of the heavy chain of said immunologic molecule comprises amino acids 26 to 32 of SEQ ID NO: 80;    (e) an immunologic molecule, wherein the CDR2 region of the heavy chain of said immunologic molecule comprises amino acids 53 to 56 of SEQ ID NO: 80; and    (f) an immunologic molecule, wherein the CDR3 region of the heavy chain of said immunologic molecule comprises amino acids 100 to 107 of SEQ ID NO: 80.    
     
     
         11 . The monoclonal antibody of  claim 5 , wherein said monoclonal antibody is 77A3.  
     
     
         12 . The monoclonal antibody of  claim 5 , wherein said monoclonal antibody is 49C9.  
     
     
         13 . The monoclonal antibody of  claim 5 , wherein said monoclonal antibody is 70B11.  
     
     
         14 . A method of making the monoclonal antibody of  claim 5  comprising: 
 (a) immunizing an animal with α2-antiplasmin or fragment thereof;  
 (b) fusing cells from the animal with tumor cells to make a hybridoma cell line;  
 (c) cloning the hybridoma cell line;  
 (d) selecting for the monoclonal antibody capable of binding to both (1) human and nonhuman circulating α2-antiplasmins and (2) human and nonhuman fibrin crosslinked α2-antiplasmins; and  
 (e) obtaining the monoclonal antibody.  
 
     
     
         15 . A hybridoma cell line which produces the monoclonal antibody of  claim 5 .  
     
     
         16 . The hybridoma cell line of  claim 15 , wherein said hybridoma cell line is ATCC Accession No. HB-12192.  
     
     
         17 . A method of making the hybridoma cell line of  claim 15  comprising: 
 (a) immunizing an animal with α2-antiplasmin or fragment thereof;  
 (b) fusing the cells from the animal with tumor cells to make the hybridoma cell line; and  
 (c) obtaining the hybridoma cell line which produces the monoclonal antibody capable of binding to both (1) human and nonhuman circulating α2-antiplasmins and (2) human and nonhuman fibrin crosslinked α2-antiplasmins.  
 
     
     
         18 . A nucleic acid molecule, selected from the group consisting of: 
 (a) a nucleic acid molecule comprising a nucleotide sequence encoding for amino acids 1 to 107 of SEQ ID NO: 5;    (b) a nucleic acid molecule comprising a nucleotide sequence encoding for amino acids 1 to 107 of SEQ ID NO: 7;    (c) a nucleic acid molecule comprising a nucleotide sequence encoding for amino acids 1 to 107 of SEQ ID NO: 9;    (d) a nucleic acid molecule comprising a nucleotide sequence encoding for amino acids 1 to 107 of SEQ ID NO: 75;    (e) a nucleic acid molecule comprising a nucleotide sequence encoding for amino acids 1 to 119 of SEQ ID NO: 11;    (f) a nucleic acid molecule comprising a nucleotide sequence encoding for amino acids 1 to 119 of SEQ ID NO: 13;    (g) a nucleic acid molecule comprising a nucleotide sequence encoding for amino acids 1 to 119 of SEQ ID NO: 15; and    (h) a nucleic acid molecule comprising a nucleotide sequence encoding for amino acids 1 to 119 of SEQ ID NO: 79.    
     
     
         19 . A method for treating pulmonary embolism, myocardial infarction, or thrombosis in a patient comprising administering a therapeutically effective amount of an immunologic molecule of  claim 1  to said patient.  
     
     
         20 . The method of  claim 19 , wherein said immunologic molecule is a monoclonal antibody.  
     
     
         21 . The method of  claim 20 , wherein said monoclonal antibody is 77A3.  
     
     
         22 . The method of  claim 19 , wherein said immunologic molecule is administered by continuous intravenous infusion or by bolus.  
     
     
         23 . A method of treatment for pulmonary embolism, myocardial infarction, or thrombosis in a patient which comprises co-administering to a patient in need of such treatment: 
 (a) a therapeutically effective amount of an immunologic molecule of  claim 1;  and    (b) a therapeutically effective amount of a thrombolytic agent, wherein said immunologic molecule (a) is different from said thrombolytic agent (b), thereby treating said patient.    
     
     
         24 . The method of  claim 23 , wherein said immunologic molecule is a monoclonal antibody.  
     
     
         25 . The method of  claim 24 , wherein said monoclonal antibody is 77A3.  
     
     
         26 . The method of  claim 23 , wherein said thrombolytic agent is plasmin.  
     
     
         27 . The method of  claim 23 , wherein said thrombolytic agent is an anti-coagulant which inhibits fibrin.  
     
     
         28 . The method of  claim 27 , wherein said anti-coagulant is selected from the group consisting of heparin, hirudin and activated protein C.  
     
     
         29 . The method of  claim 23 , wherein said thrombolytic agent is an anti-coagulant which inhibits platelets.  
     
     
         30 . The method of  claim 23 , wherein said thrombolytic agent is a plasminogen activator.  
     
     
         31 . The method of  claim 30 , wherein said plasminogen activator is selected from the group consisting of streptokinase, prourokinase, urokinase, tissue-type plasminogen activator, staphylokinase, and vampire bat plasminogen activator.  
     
     
         32 . The method of  claim 23 , wherein both said immunologic molecule (a) and said thrombolytic agent (b) are provided to said patient by an intravenous infusion or by an intravenously injected bolus.  
     
     
         33 . The method of  claim 23 , wherein said patient is provided with a first bolus containing said immunologic molecule (a) and a subsequently administered second bolus containing said thrombolytic agent (b).  
     
     
         34 . The method of  claim 23 , wherein: 
 (1) said immunologic molecule (a) is provided to said patient at a dose of between 3 to 600 nmole per kg of patient weight; and    (2) said thrombolytic agent (b) is provided to said patient at a dose of between 0.01 to 3.0 mg per kg of patient weight.    
     
     
         35 . A kit useful for carrying out the method of  claim 23 , being compartmentalized in close confinement to receive two or more container means therein, which comprises: 
 (1) a first container containing a therapeutically effective amount of said immunologic molecule (a); and    (2) a second container containing a therapeutically effective amount of said thrombolytic agent (b), wherein said immunologic molecule (a) is different from said thrombolytic agent (b).