Human coagulation factor VII polypeptides

Abstract

The present invention relates to novel human coagulation Factor VIIa variants having coagulant activity as well as polynucleotide constructs encoding such variants, vectors and host cells comprising and expressing the polynucleotide, pharmaceutical compositions, uses and methods of treatment.

1 . A polynucleotide construct encoding a Factor VII polypeptide, wherein said polypeptide comprises at least two substitutions relative to the amino acid sequence of SEQ ID NO:1 and wherein said substitutions comprise: (i) a first substitution consisting of replacement of F374 with any other amino acid, and (ii) one or more substitutions consisting of replacement with any other amino acid of one or more amino acids selected from the group consisting of L305, S314, K157, K337, D334, S336, V158, E296, and M298.  
     
     
         2 . The polynucleotide construct according to  claim 1 , wherein said construct is a vector.  
     
     
         3 . A host cell comprising the polynucleotide construct according to  claim 1 .  
     
     
         4 . The host cell according to  claim 3 , which is a eukaryotic cell.  
     
     
         5 . The host cell according to  claim 4 , which is of mammalian origin.  
     
     
         6 . The host cell according to  claim 5 , wherein the cell is selected from the group consisting of CHO cells, HEK cells and BHK cells.  
     
     
         7 . A transgenic animal containing and expressing the polynucleotide construct as defined in  claim 1 .  
     
     
         8 . A transgenic plant containing and expressing the polynucleotide construct as defined in  claim 1 .  
     
     
         9 . A method for producing the Factor VII polypeptide, the method comprising (i) cultivating a cell as defined in  claim 3  in an appropriate growth medium under conditions allowing expression of the polynucleotide construct and (ii) recovering the resulting polypeptide from the culture medium.  
     
     
         10 . A method for producing the Factor VII polypeptide, the method comprising recovering the Factor VII polypeptide from milk produced by the transgenic animal defined in  claim 7 .  
     
     
         11 . A method for producing the Factor VII polypeptide, the method comprising (i) cultivating a cell of a transgenic plant as defined in  claim 8 , and (ii) recovering the Factor VII polypeptide from the plant.  
     
     
         12 . The host cell according to  claim 2 , wherein the cell is selected from the group consisting of CHO cells, HEK cells and BHK cells.