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 . An isolated polynucleotide construct encoding a Factor VII variant, wherein said variant is a polypeptide comprising at least one amino acid substitution relative to the amino acid sequence of SEQ ID NO:1, wherein said substitution is replacement with any other amino acid of one or more amino acids selected from the group consisting of amino acids, 313, 314, 317-329, 364, 366, 373, and 376 of SEQ ID NO:1. 
     
     
         1 . The polynucleotide construct as defined in  claim 1 , wherein S314 is replaced with any other amino acid. 
     
     
         3 . The polynucleotide construct as defined in  claim 1 , wherein the polypeptide further comprises an additional replacement with any other amino acid of at least one amino acid in the remaining positions in the protease domain. 
     
     
         4 . The polynucleotide construct as defined in  claim 3 , wherein at the most 20 additional amino acids in the remaining positions in the protease domain have been replaced with any other amino acids. 
     
     
         5 . The polynucleotide construct as defined in  claim 4 , wherein at least one amino acid corresponding to an amino acid at a position selected from 330-339 of SEQ ID NO:1 has been replaced with any other amino acid. 
     
     
         6 . A polynucleotide construct as defined in  claim 5 , wherein the additional replacement comprises replacement of one or more amino acids selected from the group consisting of amino acids L305, K157, K337, D334, S336, V158, E296, and M298 of SEQ ID NO:1. 
     
     
         7 . A polynucleotide construct as defined in  claim 1 , wherein S314 has been replaced by an amino acid selected from the group consisting of Gly and Glu. 
     
     
         8 . A polynucleotide construct as defined in  claim 1 , wherein said S314 has been replaced by Glu. 
     
     
         9 . A polynucleotide construct as defined in  claim 1 , wherein said polypeptide further comprises replacement of K316 by Gin. 
     
     
         10 . A polynucleotide construct as defined in  claim 1 , wherein the polypeptide is selected from the group consisting of S314E/K316H-FVII and S314E/K316Q-FVII. 
     
     
         11 . A polynucleotide as defined in  claim 1 , wherein said construct is a vector. 
     
     
         12 . A host cell comprising a polynucleotide construct as defined in  claim 11 . 
     
     
         13 . A host cell as defined in  claim 12 , wherein said cell is a eukaryotic cell. 
     
     
         14 . A host cell as defined in  claim 13 , wherein said cell is of mammalian origin. 
     
     
         15 . A host cell as defined in  claim 14 , wherein the cell is selected from the group consisting of CHO cells, HEK cells, and BHK cells. 
     
     
         16 . A method for producing a Factor VII polypeptide, the method comprising (i) cultivating a cell as defined in 12 in an appropriate growth medium under conditions allowing expression of the polynucleotide construct and (ii) recovering the resulting polypeptide from the culture.