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 method of treating a blood coagulation disorder, comprising administering to a subject in need thereof an effective amount of a glycoprotein and an effective amount of a Factor VIII (FVIII),
wherein the glycoprotein comprises a truncated von Willebrand Factor (VWF) capable of binding to the FVIII,
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,
wherein the glycoprotein is administered intravenously or subcutaneously, and
wherein the FVIII is administered intravenously or subcutaneously.
2. The method according to claim 1 , wherein the mean residence time (MRT) of the FVIII is increased and/or the clearance of the FVIII is decreased by co-administration of the glycoprotein, as compared to a treatment with FVIII alone; and/or wherein the frequency of administration of the FVIII is reduced as compared to a treatment with FVIII alone.
3. The method of claim 1 , wherein the glycoprotein and the FVIII are administered simultaneously.
4. The method of claim 1 , wherein the glycoprotein and the FVIII are administered separately and sequentially.
5. The method of claim 1 , wherein at least 70% of the N-glycans comprise at least one α-2,6-sialic acid moiety or at least one α-2,3-sialic acid moiety.
6. The method of claim 1 , wherein at least 80% of the N-glycans comprise at least one sialic acid moiety.
7. The method of claim 1 , wherein at least 85% of the N-glycans comprise at least one sialic acid moiety.
8. The method 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.
9. The method 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.
10. The method 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.
11. The method 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.
12. The method of claim 1 , wherein a half-life extending heterologous polypeptide is fused to the truncated VWF, and/or a half-life extending moiety is conjugated to the glycoprotein.
13. The method of claim 1 , wherein the glycoprotein is a dimer.
14. The method of claim 13 , wherein the affinity of the dimeric glycoprotein to FVIII is greater than the affinity of the corresponding monomeric glycoprotein to FVIII.