Coagulation factor VII polypeptides
The present invention relates to novel coagulation Factor VII polypeptides, polynucleotide constructs encoding such polypeptides, as well as vectors and host cells comprising and expressing the polynucleotide, pharmaceutical compositions, uses and methods of treatment.
1. An isolated Factor VII variant polypeptide comprising an additional N-glycosylation site N-Xaa-S/T compared to the sequence of wild-type Factor VII (SEQ ID NO:1), wherein said additional glycosylation site is an amino acid substitution at amino acid I69 of SEQ ID NO:1, wherein Xaa is any amino acid except P, and wherein said variant polypeptide exhibits decreased Tissue Factor binding affinity compared to wild-type Factor VII.
2. The isolated Factor VII variant polypeptide of claim 1 , wherein the isolated Factor VII variant polypeptide is FVII-I69N.
3. The isolated Factor VII variant polypeptide of claim 1 , wherein the dissociation constant (K d ) of said isolated Factor VII polypeptide for Tissue Factor binding is higher than 5 nM.
4. The isolated Factor VII variant polypeptide of claim 3 , wherein the dissociation constant (K d ) of said isolated Factor VII polypeptide for Tissue Factor binding is higher than 50 nM.
5. A composition comprising the isolated Factor VII variant polypeptide of claim 1 and a pharmaceutically acceptable carrier.
6. A method for treating bleeding episodes or bleeding disorders in a subject in need thereof, comprising: administering to the subject in need thereof a therapeutically effective amount of the isolated Factor VII variant polypeptide of claim 1 .
7. A method for treating bleeding episodes or bleeding disorders or for the enhancement of normal haemostatic system in a subject in need thereof, comprising:
administering to the subject in need thereof a prophylactically effective amount of the isolated Factor VII variant polypeptide of claim 1 .
8. The isolated Factor VII variant polypeptide of claim 1 , wherein said additional glycosylation site further comprise an amino acid substitution at amino acid Q64 of SEQ ID NO:1.
9. The isolated Factor VII variant polypeptide of claim 1 , wherein the N-glycosylation site is introduced at amino acid I69 of SEQ ID NO: 1.
10. The isolated Factor VII variant polypeptide of claim 1 , wherein said additional glycosylation site further comprise an amino acid substitution at amino acid F71 of SEQ ID NO:1.
11. The isolated Factor VII variant polypeptide of claim 1 , wherein said additional glycosylation site further comprise an amino acid substitution at amino acid E77 of SEQ ID NO:1.
12. The isolated Factor VII variant polypeptide of claim 1 , wherein said additional glycosylation site further comprise an amino acid substitution at amino acid G78 of SEQ ID NO:1.
13. The isolated Factor VII variant polypeptide of claim 1 , wherein said additional glycosylation site further comprise an amino acid substitution at amino acid Q88 of SEQ ID NO:1.
14. The isolated Factor VII variant polypeptide of claim 1 , wherein said additional glycosylation site further comprise an amino acid substitution at amino acid F275 of SEQ ID NO:1.
15. The isolated Factor VII variant polypeptide of claim 1 , wherein said additional glycosylation site further comprise an amino acid substitution at amino acid M306 of SEQ ID NO:1.
16. The isolated Factor VII variant polypeptide of claim 1 , wherein said additional glycosylation site further comprise an amino acid substitution at amino acid T307 of SEQ ID NO:1.
17. The isolated Factor VII variant polypeptide of claim 1 , wherein said additional glycosylation site further comprise an amino acid substitution at amino acid D309 of SEQ ID NO:1.
18. The isolated Factor VII variant polypeptide of claim 1 , wherein the isolated Factor VII variant polypeptide is FVII-(I69N/F71T).
19. The isolated Factor VII variant polypeptide of claim 1 , wherein the isolated Factor VII variant polypeptide is FVII-(F71N/L73T).
20. The isolated Factor VII variant polypeptide of claim 1 , wherein the isolated Factor VII variant polypeptide is FVII-(D309N/L311T).