Human Coagulation Factor VII Variants

Abstract

The invention concerns novel coagulation factor VII variants, wherein the Phe residue in position 374 of SEQ ID NO 1 has been replaced by another amino acid residue which can be encoded by nucleic acid constructs and, optionally, wherein at least one other amino acid residue in the remaining positions in the protease domain has been replaced by another amino acid residue which can be encoded by nucleic acid constructs; with the proviso that the variant is not FVII(Ala305).

1 . A nucleic acid construct comprising a nucleotide sequence encoding a human coagulation Factor VII variant, wherein said variant comprises a substitution of the Leu in position 305 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 and 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. 
     
     
         2 . A recombinant vector comprising a nucleic acid construct as defined in  claim 1 . 
     
     
         3 . A recombinant host cell comprising a nucleic acid construct as defined in  claim 1 . 
     
     
         4 . A recombinant host cell as defined in  claim 3 , wherein the cell is of mammalian origin. 
     
     
         5 . A recombinant host cell as defined in  claim 4 , wherein the cell is selected from the group consisting of CHO cells and BHK cells. 
     
     
         6 . A transgenic animal comprising the nucleic acid construct defined in  claim 1 . 
     
     
         7 . A transgenic plant comprising the nucleic acid construct defined in  claim 1 . 
     
     
         8 . A method for producing a human coagulation Factor VII variant, which comprises (i) cultivating a cell as defined in  claim 3  in an appropriate growth medium under conditions allowing expression of the nucleic acid construct and (ii) recovering the resulting polypeptide from the culture medium. 
     
     
         9 . A method for producing a human coagulation Factor VII variant, which comprises recovering the variant from milk produced by a transgenic animal as defined in  claim 6 . 
     
     
         10 . A method for producing a human coagulation Factor VII variant, comprising (i) cultivating a cell of a transgenic plant as defined in  claim 7  under conditions in which the variant is expressed, and (ii) recovering the variant from the resulting plant.