Human Coagulation Factor VII Variants

Abstract

The present invention encompasses isolated human coagulation Factor VII variants comprising a substitution of Phe in position 374 of SEQ ID NO 1 with another amino acid residue.

1 . An isolated human coagulation Factor VII variant comprising a substitution of Phe in position 374 of SEQ ID NO 1 with an amino acid residue selected from the group consisting of Val, Ile, Met, Phe, Trp, Pro, Gly, Ser, Thr, Cys, Tyr, Asn, Glu, Lys, Arg, His, Asp and Gln. 
     
     
         2 . A Factor VII variant as defined in  claim 1 , wherein the substituted amino acid is Pro. 
     
     
         3 . A Factor VII variant as defined in  claim 1 , further comprising a second substitution selected from the group consisting of (i) position 274; (ii) any of positions 300-304; (iii) any of positions 306-312; and (iv) combinations of any of the foregoing. 
     
     
         4 . A Factor VII variant as defined in  claim 3 , wherein the second substitution is at position 274. 
     
     
         5 . A Factor VII variant as defined in  claim 3 , wherein the second substitution is at any of positions 300-304. 
     
     
         6 . A Factor VII variant as defined in  claim 3 , wherein the second substitution is at any of positions 306-312. 
     
     
         7 . A Factor VII variant as described in  claim 1 , wherein the Phe residue in position 374 is the only amino acid residue that has been replaced relative to the sequence of SEQ ID NO:1. 
     
     
         8 . A Factor VII variant as defined in  claim 1 , wherein the ratio between the activity of the variant and the activity of native Factor VII polypeptide having a sequence shown in SEQ ID NO 1 is at least about 1.25 when tested in an in vitro hydrolysis assay. 
     
     
         9 . A Factor VII variant as defined in  claim 8 , wherein the ratio is at least about 2.0, 
     
     
         10 . A Factor VII variant as defined in  claim 9 , wherein the ratio is at least about 4.0. 
     
     
         11 . An isolated nucleic acid construct comprising a nucleotide sequence encoding a Factor VII variant as defined in  claim 1 . 
     
     
         12 . A recombinant vector comprising a nucleic acid construct as defined in  claim 11 . 
     
     
         13 . A recombinant host cell comprising a nucleic acid construct as defined in  claim 11 . 
     
     
         14 . A recombinant host cell as defined in  claim 13 , wherein the cell is of mammalian origin. 
     
     
         15 . A recombinant host cell as defined in  claim 14 , wherein the cell is selected from the group consisting of CHO cells and BHK cells. 
     
     
         16 . A method for producing a human coagulation Factor VII variant, which comprises (i) cultivating a cell as defined in  claim 13  in an appropriate growth medium under conditions allowing expression of the nucleic acid construct and (ii) recovering the resulting polypeptide from the culture medium. 
     
     
         17 . A pharmaceutical composition comprising (i) a human coagulation Factor VII variant as defined in  claim 1  and (ii) a pharmaceutically acceptable carrier or excipient. 
     
     
         18 . A method for the treatment of bleeding episodes in a subject or for the enhancement of the normal haemostatic system, the method comprising administering to a subject in need of such treatment a therapeutically or prophylactically effective amount of a human coagulation Factor VII variant as defined in  claim 1 .