Novel human kunitz-type inhibitors and methods relating thereto

Abstract

The present invention provides isolated DNA molecules comprising a DNA segment encoding novel human Kunitz-type inhibitors. Also provided are DNA constructs comprising a first DNA segment encoding a novel human Kunitz-type inhibitor wherein said first DNA segment is operably linked to additional DNA segments required for the expression for the first DNA segment, as well as host cells containing such DNA constructs and methods for producing proteins from the host cells.

1 . An isolated human Kunitz-type inhibitor that inhibits plasmin in a mammal and wherein DNA sequence encoding the human Kunitz-type inhibitor hybridizes to nucleotides 138-305 of SEQ ID NO:1 under highly stringent hybridization conditions.  
     
     
         2 . The isolated human Kunitz-type inhibitor of  claim 1  wherein any differences between the human Kunitz-type inhibitor and amino acid number 34 to amino acid number 89 of SEQ ID NO:2 are due to conservative amino acid substitutions.  
     
     
         3 . A pharmaceutical composition comprising the human Kunitz-type inhibitor of  claim 1 .  
     
     
         4 . The pharmaceutical composition of  claim 3  wherein the human Kunitz-type inhibitor is isolated from  E. coli.    
     
     
         5 . A DNA construct comprising a first DNA segment, wherein the first DNA segment is the DNA sequence of  claim 1 , operably linked to additional DNA segments required for the expression of the first DNA segment.  
     
     
         6 . A host cell comprising the DNA construct of  claim 5  wherein the host cell expresses the human Kunitz-type inhibitor encoded by the first DNA segment.  
     
     
         7 . The host cell of  claim 6  wherein the host cell is  E. coli.    
     
     
         8 . A method for producing human Kunitz-type inhibitor comprising: 
 culturing a cell according to  claim 6;  and    isolating the human Kunitz-type inhibitor produced by the cell.    
     
     
         9 . The method of  claim 8  wherein the cell is  E. coli.    
     
     
         10 . An isolated DNA sequence that hybridizes to nucleotides 138-305 SEQ ID NO:1 under highly stringent hybridization conditions, wherein the isolated DNA sequence encodes a human Kunitz-type inhibitor that inhibits plasmin in a mammal.  
     
     
         11 . The isolated DNA sequence of  claim 10  wherein any differences between the encoded human Kunitz-type inhibitor and amino acid number 34 to amino acid number 89 of SEQ ID NO:2 are due to conservative amino acid substitutions.  
     
     
         12 . An isolated human Kunitz-type inhibitor that inhibits plasmin in a mammal and wherein DNA sequence encoding the human Kunitz-type inhibitor hybridizes to nucleotides 39-743 of SEQ ID NO:1 under highly stringent hybridization conditions.  
     
     
         13 . The isolated human Kunitz-type inhibitor of  claim 12  wherein any differences between the human Kunitz-type inhibitor and amino acid number 1 to amino acid number 235 of SEQ ID NO:2 are due to conservative amino acid substitutions.  
     
     
         14 . A pharmaceutical composition comprising the human Kunitz-type inhibitor of  claim 12 .  
     
     
         15 . The pharmaceutical composition of  claim 14  wherein the human Kunitz-type inhibitor is isolated from  E. coli.    
     
     
         16 . A DNA construct comprising a first DNA segment, wherein the first DNA segment is the DNA sequence of  claim 12 , operably linked to additional DNA segments required for the expression of the first DNA segment.  
     
     
         17 . A host cell comprising the DNA construct of  claim 16  wherein the host cell expresses the human Kunitz-type inhibitor encoded by the first DNA segment.  
     
     
         18 . The host cell of  claim 17  wherein the host cell is  E. coli.    
     
     
         19 . A method for producing human Kunitz-type inhibitor comprising: 
 culturing a cell according to  claim 17;  and    isolating the human Kunitz-type inhibitor produced by the cell.    
     
     
         20 . The method of  claim 19  wherein the cell is  E. coli.    
     
     
         21 . An isolated DNA sequence that hybridizes to nucleotides 39-743 SEQ ID NO: 1 under highly stringent hybridization conditions, wherein the isolated DNA sequence encodes a human Kunitz-type inhibitor that inhibits plasmin in a mammal.  
     
     
         22 . The isolated DNA sequence of  claim 21  wherein any differences between the encoded human Kunitz-type inhibitor and amino acid number 1 to amino acid number 235 of SEQ ID NO:2 are due to conservative amino acid substitutions.