Glycopegylated factor IX

• US 8,632,770 B2
• Filed: 08/06/2010
• Issued: 01/21/2014
• Est. Priority Date: 12/03/2003
• Status: Active Grant

• Current Assignee: Novo Nordisk A/S
• Original Assignee: Novo Nordisk A/S
• Inventors: DeFrees, Shawn, Bayer, Robert J., Bowe, Caryn, Panneer-Selvam, Krishnasamy
• Primary Examiner(s): ROBINSON, HOPE A
• Application Number:
  US12/851,651
• Publication Number:
  US 20100330060A1
• Time in Patent Office: 1,264 Days
• Field of Search: None
• US Class Current: 424/94.3
• CPC Class Codes: A61K 38/4846 Factor VII (3.4.21.21); Factor IX (3.4.21.22); Factor Xa (3.4.21.6); Factor XI (3.4.21.27); Factor XII (3.4.21.38)
• CPC Class Codes: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/04 Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present invention provides conjugates between Factor IX and PEG moieties. The conjugates are linked via an intact glycosyl linking group interposed between and covalently attached to the peptide and the modifying group. The conjugates are formed from glycosylated peptides by the action of a glycosyltransferase. The glycosyltransferase ligates a modified sugar moiety onto a glycosyl residue on the peptide. Also provided are methods for preparing the conjugates, methods for treating various disease conditions with the conjugates, and pharmaceutical formulations including the conjugates.

Claims

1. A Factor IX peptide conjugate comprising a glycosyl linking group attached to an amino acid residue of a Factor IX peptide comprising SEQ ID NO:

1, wherein the glycosyl linking group is attached to an amino acid of SEQ ID NO;

1 selected from the group consisting of Asn 157 and Asn 167, the glycosyl linking group comprising a modified sialyl residue having the formula;

2. The Factor IX peptide conjugate according to claim 1, wherein the moiety:

3. The Factor IX peptide conjugate according to claim 2, wherein said moiety has a formula that is a member selected from:

4. The Factor IX peptide conjugate according to claim 1, wherein said glycosyl linking group has a formula selected from:

5. The Factor IX peptide conjugate according to claim 4, wherein said glycosyl linking group has the formula:

6. The Factor IX peptide conjugate according to claim 5, wherein said glycosyl linking group attached to said amino acid residue has the formula:

7. The Factor IX peptide conjugate according to claim 1, wherein said peptide comprises at least one glycosyl linking group comprising a substructure having the formula:

8. The Factor IX peptide according to claim 7, wherein said at least one glycosyl linking group attached to an amino acid of said peptide has a formula selected from:

9. A pharmaceutical formulation comprising the Factor IX peptide conjugate of claim 1, and a pharmaceutically acceptable carrier.

18. The Factor IX peptide conjugate according to claim 1, wherein

R² is COOR⁷,

wherein R⁷ is H;

R³ is H;

R⁴ is OR⁸, wherein

R⁸ is H;

L^a is NHC(O)CH₂NHC(O)OCH₂

X⁵ is H; and

R¹⁶ and R¹⁷ are poly(ethylene glycol) having a total molecular weight of about 40 kDa.

20. The Factor IX peptide conjugate according to claim 1, wherein

R² is COOR⁷,

wherein R⁷ is H;

R³ is H;

R⁴ is OR⁸, wherein

R⁸ is H;

L^a is NHC(O)CH₂NHC(O)OCH₂

X⁵ is H; and

R¹⁶ and R¹⁷ are poly(ethylene glycol) having a total molecular weight of about 40 kDa.

10. A Factor IX peptide conjugate comprising a glycosyl linking group attached to an amino acid residue of a Factor IX peptide comprising SEQ ID NO:

1, wherein the glycosyl linking group is attached to an amino acid of SEQ ID NO;

1 selected from the group consisting of Asn 157 and Asn 167, the glycosyl linking group comprising a modified sialyl residue having the formula;

11. The Factor IX peptide conjugate according to claim 10, wherein said modified sialyl residue has the formula:

12. The Factor IX peptide conjugate according to claim 11, wherein Q is selected from H and CH₃.

13. The Factor IX peptide conjugate according to claim 10, wherein said peptide comprises at least one glycosyl linking group comprising a substructure having the formula:

14. The Factor IX peptide conjugate according to claim 13, wherein said at least one glycosyl linking group attached to an amino acid of said peptide has a formula selected from:

15. The peptide conjugate according to claim 10, wherein said glycosyl linking group comprises the formula:

16. The peptide conjugate according to claim 15, wherein s is 1;

and f is an integer from about 200 to about 300.

17. A pharmaceutical formulation comprising the Factor IX peptide conjugate according to claim 10, and a pharmaceutically acceptable carrier.

19. A pharmaceutical formulation comprising the Factor IX peptide conjugate according to claim 10, and a pharmaceutically acceptable carrier.