Proteolytically cleavable fusion proteins with high molar specific activity

1. A fusion protein comprising:

a) a coagulation factor,

b) a half-life enhancing polypeptide (HLEP) wherein the half-life enhancing polypeptide is, and

c) a peptide linker which joins the coagulation factor and the half-life enhancing polypeptide;

wherein the peptide linker is cleavable by proteases involved in coagulation, and wherein the fusion protein has, in comparison to a respective fusion protein linked by a non-cleavable linker having the amino acid sequence GGGGGGV (SEQ ID NO: 94), at least one of the following properties:

i) an increased molar specific activity in at least one coagulation-related assay,

ii) an increased inactivation rate of the activated coagulation factor after the peptide linker is proteolytically cleaved in a coagulation-related mode, and

iii) an increased elimination rate of the activated coagulation factor after the peptide linker is proteolytically cleaved in a coagulation-related mode.

2. The fusion protein of claim 1 , wherein said fusion protein has a higher in vivo recovery compared to the in vivo recovery of the coagulation factor when it is not fused to a half-life enhancing polylpeptide.

3. The fusion protein of claim 1 , wherein said fusion protein has an increased half-life in plasma compared to the half-life in plasma of the coagulation factor when it is not fused to a half-life enhancing polylpeptide.

4. The fusion protein of claim 1 wherein the coagulation factor is a vitamin-K dependent coagulation factor.

5. The fusion protein according of claim 1 wherein the coagulation factor is FIX.

6. The fusion protein of claim 1 wherein the peptide linker is cleavable by FXIa and/or FVIIa/TF.

7. The fusion protein of claim 1 , wherein the molar specific activity of the fusion protein is increased at least 25% compared to that of the respective fusion protein linked by a non-cleavable linker having the amino acid sequence GGGGGGV (SEQ ID NO: 94) in at least one coagulation-related assay.

8. The fusion protein of claim 1 , wherein the inactivation rate of the coagulation factor after cleavage of the peptide linker which links the coagulation factor to the half-life enhancing polypeptide is increased by at least 10% as compared to the inactivation rate of the coagulation factor in the corresponding fusion protein linked by a non-cleavable linker having the amino acid sequence GGGGGGV (SEQ ID NO: 94).

9. The fusion protein of claim 1 , wherein the elimination rate of the coagulation factor after cleavage of the peptide linker which links the coagulation factor to the half-life enhancing polypeptide is increased by at least 10% as compared to the elimination rate of the coagulation factor in the corresponding fusion protein linked by a non-cleavable linker having the amino acid sequence GGGGGGV (SEQ ID NO: 94).

10. The fusion protein of claim 1 , wherein the linker is cleavable by a protease that naturally activates the coagulation factor in vivo.

11. The fusion protein of claim 10 , wherein the kinetics of the linker cleavage by the protease is not delayed by more than a factor of 3 compared to the kinetics of the activation of said coagulation factor.

12. The fusion protein of claim 1 , wherein the linker is cleavable by a protease that is naturally activated in vivo by the coagulation factor.

13. The fusion protein of claim 1 , wherein the linker is cleavable by FXIa and/or by FVIIaITF, and wherein the coagulation factor is FIX.

14. The fusion protein of claim 1 wherein the linker is cleavable by FXa and/or FVIIa/TF, and wherein the coagulation factor is FVIIa.

15. The fusion protein of claim 1 , wherein the linker comprises SEQ ID NO:113.

16. A polynucleotide encoding the fusion protein of claim 1 .

17. A plasmid or vector comprising the polynucleotide of claim 16 .

18. A plasmid or vector according to claim 17 , which is an expression vector.

19. A plasmid or vector according to claim 17 , wherein the vector is a transfer vector for use in human gene therapy.

20. A host cell comprising a polynucleotide according to claim 16 .

21. A method of producing a fusion protein of claim 1 , comprising culturing host cells comprising a polynucleotide encoding the fusion protein under conditions such that the fusion protein is expressed.

22. A pharmaceutical composition comprising

(a) the fusion protein of claim 1 ,

(b) a polynucleotide encoding said fusion protein, or

(c) a plasmid or vector comprising a polynucleotide encoding said fusion protein.

23. A method of administering an effective amount of the fusion protein of claim 1 to a patient in need thereof, comprising

(a) administering said fusion protein.

24. The method of claim 23 , wherein the patient suffers from a blood coagulation disorder.

25. The method of claim 24 , wherein the blood coagulation disorder is hemophilia B.

26. The method of claim 24 , wherein the blood coagulation disorder is FVII and/or FVIIa deficiency.

27. The method according to claim 24 , wherein the blood coagulation disorder is hemophilia A.

28. The method according to claim 24 , wherein the fusion protein is effective to act in the patient as a procoagulant.

29. The fusion protein of claim 1 , wherein the coagulation factor, albumin, or immunoglobulin comprises a sequence that is 95% identical to the sequence of a wild-type human coagulation factor, human serum albumin, or a wild-type human immunoglobulin.

30. The method of claim 21 further comprising recovering the fusion protein from the host cells or from the culture medium.

31. The method of claim 24 , wherein the administration comprises administering a composition comprising the fusion protein of claim 1 .