Methods for preparing modified von Willebrand Factor
The present invention provides modified von Willebrand Factor molecules, methods for their preparation and uses thereof. The invention further provides pharmaceutical compositions for treating coagulation disorders.
1. A glycoprotein comprising a truncated von Willebrand Factor (VWF), wherein the truncated VWF is capable of binding to a Factor VIII (FVIII), and wherein the glycoprotein comprises N-glycans, wherein i) at least 75% of the N-glycans comprise at least one sialic acid moiety, ii) less than 35% of the N-glycans comprise two or more terminal and non-sialylated galactose residues, and/or iii) less than 6% of the N-glycans comprise three or more terminal and non-sialylated galactose residues.
2. The glycoprotein of claim 1 , wherein at least 70% of the N-glycans comprise at least one a-2,6-sialic acid moiety or at least one a-2,3-sialic acid moiety.
3. The glycoprotein of claim 1 , wherein at least 80% of the N-glycans comprise at least one sialic acid moiety.
4. The glycoprotein of claim 1 , wherein at least 85% of the N-glycans comprise at least one sialic acid moiety.
5. The glycoprotein of claim 1 , wherein the truncated VWF comprises (a) amino acids 776 to 805 of SEQ ID NO:9, or (b) an amino acid sequence having at least 90% sequence identity to amino acids 776 to 805 of SEQ ID NO:9.
6. The glycoprotein of claim 1 , wherein the truncated VWF consists of (a) amino acids 776 to 805 of SEQ ID NO:9, or (b) an amino acid sequence having at least 90% sequence identity to amino acids 764 to 1242 of SEQ ID NO:9.
7. The glycoprotein of claim 1 , wherein the truncated VWF comprises (a) amino acids 764 to 1242 of SEQ ID NO:9, or (b) an amino acid sequence having at least 90% sequence identity to amino acids 764 to 1242 of SEQ ID NO:9.
8. The glycoprotein of claim 1 , wherein the truncated VWF consists of (a) amino acids 764 to 1242 of SEQ ID NO:9, or (b) an amino acid sequence having at least 90% sequence identity to amino acids 764 to 1242 of SEQ ID NO:9.
9. The glycoprotein of claim 1 , further comprising a half-life extending heterologous polypeptide fused to the truncated VWF, and/or a half-life extending moiety is conjugated to the glycoprotein.
10. The glycoprotein of claim 1 , wherein the glycoprotein is a dimer.
11. The glycoprotein of claim 10 , wherein the affinity of the dimeric glycoprotein to FVIII is greater than the affinity of a corresponding monomeric glycoprotein to FVIII.
12. A pharmaceutical composition comprising the glycoprotein of claim 1 and a pharmaceutically acceptable excipient.
13. The pharmaceutical composition of claim 12 , wherein at least 50% of the glycoprotein in the pharmaceutical composition is present as a dimer.
14. A pharmaceutical kit comprising (i) a FVIII and (ii) the glycoprotein of claim 1 .
15. The pharmaceutical kit of claim 14 , wherein at least 50% of the glycoprotein in the pharmaceutical kit is present as a dimer.