Reversibly inactivated acidified plasmin

Abstract

The present invention provides a fibrinolytic composition useful as a therapeutic for administration to a patient having a thrombotic occlusion. In one aspect of the present invention, the fibrinolytic composition comprises a reversibly inactivated acidified serine protease substantially free of a plasminogen activator, a low buffering capacity buffer, and optionally, a stabilizing agent. In another aspect of the invention, the fibrinolytic composition of the present invention comprises a reversibly inactivated acidified plasmin substantially free of a plasminogen activator, a low buffering capacity buffer, and optionally, a stabilizing agent.

What is claimed is:  
     
         1 . A fibrinolytic composition comprising: 
 a reversibly inactive, acidified plasmin, the plasmin being substantially free of a plasminogen activator;    a low buffering capacity buffer; and    optionally, a stabilizing agent.    
     
     
         2 . The composition of  claim 1 , wherein the plasmin is selected from Glu-plasmin, Lys-plasmin, midi-plasmin, mini-plasmin, or micro-plasmin.  
     
     
         3 . The composition of  claim 1 , wherein the fibrinolytic composition is lyophilized.  
     
     
         4 . The composition of  claim 1 , further comprising an aqueous carrier.  
     
     
         5 . The composition of  claim 1 , further comprising an anticoagulant.  
     
     
         6 . The composition of  claim 1 , wherein the fibrinolytic composition has a pH between about 2.5 and about 4.  
     
     
         7 . The composition of  claim 1 , wherein the buffer comprises at least one acid.  
     
     
         8 . The composition of  claim 1 , wherein the buffer comprises a carboxylic acid, at least one amino acid, a derivative of the at least one amino acid, a dipeptide, an oligopeptide which includes the at least one amino acid, or a combination thereof.  
     
     
         9 . The composition of  claim 1 , wherein the buffer is selected from formic acid, acetic acid, citric acid, hydrochloric acid, lactic acid, malic acid, tartaric acid, benzoic acid, serine, threonine, methionine, glutamine, alanine, glycine, isoleucine, valine, alanine, aspartic acid, derivatives thereof, or combinations thereof.  
     
     
         10 . The composition of  claim 1 , wherein the stabilizing agent is selected from a polyhydric alcohol, a salt, citrulline, or combinations thereof.  
     
     
         11 . The composition of  claim 1 , wherein the plasmin is in the concentration range of between about 0.01 mg/ml to about 50 mg/ml.  
     
     
         12 . The composition of  claim 7 , wherein the acid is in the concentration range of between about 1 mM and about 100 mM.  
     
     
         13 . The composition of  claim 1 , wherein the stabilizing agent is a pharmaceutically acceptable carbohydrate, salt, glucosamine, thiamine, niacinamide, citrulline, or combinations thereof.  
     
     
         14 . The composition of  claim 1 , wherein the stabilizing agent is a sugar or sugar alcohol selected from glucose, maltose, mannitol, sorbitol, sucrose, lactose, trehalose, or combinations thereof.  
     
     
         15 . The composition of  claim 1 , wherein the stabilizing agent is selected from monosacchrides, disaccharides, polysaccharides, polyhydric alcohols, or combinations thereof.  
     
     
         16 . The composition of  claim 13 , wherein the carbohydrate has a concentration in the range of about 0.2% w/v to about 20% w/v.  
     
     
         17 . The composition of  claim 1 , wherein the stabilizing agent is selected from sodium chloride, potassium chloride, magnesium chloride, calcium chloride, manganese chloride or combinations thereof.  
     
     
         18 . The composition of  claim 1 , wherein the stabilizing agent is selected from a salt, glucosamine, thiamine, niacinamide, or a combination thereof and the concentration of the stabilizing agent is in the range of about 0.01M to about 1M.  
     
     
         19 . The composition of  claim 1 , wherein the buffer is present in the composition at a concentration at which the pH of the composition is raised to a physiological pH by contacting a body fluid or a thrombus.  
     
     
         20 . A fibrinolytic composition comprising: 
 a reversibly inactive, acidified serine protease, the serine protease being substantially free of a plasminogen activator;    a low buffering capacity buffer; and    optionally, a stabilizing agent.    
     
     
         21 . The composition of  claim 20 , wherein the fibrinolytic composition is lyoplilized.  
     
     
         22 . The composition of  claim 20 , further comprising an aqueous carrier.  
     
     
         23 . The composition of  claim 20 , further comprising an anticoagulant.  
     
     
         24 . The composition of  claim 20 , wherein the fibrinolytic composition has a pH between about 2.5 and about 4.  
     
     
         25 . The composition of  claim 20 , wherein the buffer comprises at least one acid.  
     
     
         26 . The composition of  claim 20 , wherein the low buffering capacity buffer comprises a carboxylic acid, at least one amino acid, a derivative of the at least one amino acid, a dipeptide, an oligopeptide which includes the at least one amino acid, or a combination thereof.  
     
     
         27 . The composition of  claim 20 , wherein the buffer is selected from acetic acid, citric acid, hydrochloric acid, lactic acid, malic acid, tartaric acid, benzoic acid, serine, threonine, methionine, glutamine, alanine, glycine, isoleucine, valine, alanine, aspartic acid, derivatives thereof, or combinations thereof.  
     
     
         28 . The composition of  claim 20 , wherein the stabilizing agent is selected from a polyhydric alcohol, a salt, citrulline, or combinations thereof.  
     
     
         29 . The composition of  claim 20 , wherein the plasmin is in the concentration range of between about 0.01 mg/ml to about 50 mg/ml.  
     
     
         30 . The composition of  claim 25 , wherein the acid is in the concentration range of between about 1 mM and about 100 mM.  
     
     
         31 . The composition of  claim 20 , wherein the stabilizing agent is a pharmaceutically acceptable carbohydrate, salt, glucosamine, thiamine, niacinamide, citrulline, or combinations thereof.  
     
     
         32 . The composition of  claim 20 , wherein the stabilizing agent is a sugar or sugar alcohol selected from glucose, maltose, mannitol, sorbitol, sucrose, lactose, trehalose, or combinations thereof.  
     
     
         33 . The composition of  claim 20 , wherein the stabilizing agent is selected from monosacchrides, disaccharides, polysaccharides, polyhydric alcohols, or combinations thereof.  
     
     
         34 . The composition of  claim 31 , wherein the carbohydrate has a concentration in the range of about 0.2% w/v to about 20% w/v.  
     
     
         35 . The composition of  claim 20 , wherein the stabilizing agent is selected from sodium chloride, potassium chloride, magnesium chloride, calcium chloride, or combinations thereof.  
     
     
         36 . The composition of  claim 20 , wherein the stabilizing agent is selected from a salt, glucosamine, thiamine, niacinamide, or a combination thereof and the concentration of the stabilizing agent is in the range of about 0.01M to about 1M.  
     
     
         37 . The composition of  claim 20 , wherein the buffer is present in the composition at a concentration at which the pH of the composition is raised to a neutral pH by adding no more than about an equal volume of serum to the composition.  
     
     
         38 . The composition of  claim 20 , wherein the serine protease is selected from trypsin, chymotrypsin, pancreatic elastase II, cathepsin G, prostate-specific antigen, leukocyte elastase, chymase, tryptase, acrosin, human tissue kallikrein, and plasmin.  
     
     
         39 . The composition of  claim 38 , wherein the plasmin is a truncated variant thereof selected from midi-plasmin, mini-plasmin, or micro-plasmin.