Glycopegylated interferon α

• US 8,268,967 B2
• Filed: 09/12/2005
• Issued: 09/18/2012
• Est. Priority Date: 09/10/2004
• Status: Active Grant

• Current Assignee: Novo Nordisk A/S
• Original Assignee: Novo Nordisk A/S
• Inventors: DeFrees, Shawn, Zopf, David A., Clausen, Henrik, Xing, Ruye
• Primary Examiner(s): Landsman, Robert
• Assistant Examiner(s): Hissong, Bruce D
• Application Number:
  US11/659,942
• Publication Number:
  US 20090028822A1
• Time in Patent Office: 2,563 Days
• Field of Search: None
• US Class Current: 530/351
• CPC Class Codes: A61K 38/212 IFN-alpha
• 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 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic 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 IFN-α conjugates including IFN-α peptides and modifying groups such as PEG moieties. The IFN-α peptide and modifying group are linked via an intact glycosyl linking group interposed between and covalently attached to the IFN-α peptide and the modifying group. The IFN-α conjugates are formed from glycosylated peptides by the action of a glycosyltransferase. The glycosyltransferase ligates a modified sugar onto an amino acid or a glycosyl residue on the IFN-α peptide. Also provided are methods for preparing the IFN-α conjugates, methods for treating various disease conditions with the IFN-α conjugates, and pharmaceutical formulations including the IFN-α conjugates.

Claims

1. An IFN-α

conjugate comprising;

an IFN-α

peptide; and

a glycosyl group attached to an asparagine, lysine, serine, or threonine residue of said IFN-α

peptide, said glycosyl group comprising a modified sialyl residue having the formula;

2. The IFN-α

conjugate according to claim 1, wherein

3. The IFN-α

conjugate according to claim 2, wherein

4. The IFN-α

conjugate according to claim 1, wherein said glycosyl group having a formula that is a member selected from;

5. The IFN-α

conjugate according to claim 1, wherein said glycosyl group having the formula;

6. The IFN-α

conjugate according to claim 5, wherein said glycosyl group attached to said amino acid residue has the formula;

7. The IFN-α

conjugate according to claim 1 wherein said glycosyl group has the formula which is a member selected from;

8. The IFN-α

conjugate according to claim 1 wherein said glycosyl group has the formula which is a member selected from;

9. The IFN-α

conjugate according to claim 8 wherein said glycosyl group has the formula which is a member selected from;

10. The IFN-α

conjugate according to claim 1, wherein said amino acid residue is a member selected from asparagine and lysine.

11. The IFN-α

conjugate according to claim 1, wherein said amino acid residue is a member selected from serine and threonine.

12. The IFN-α

conjugate according to claim 1, wherein said peptide comprises the amino acid sequence of SEQ ID NO;

1 or SEQ ID NO;

2.

14. The IFN-α

conjugate according to claim 1, wherein said IFN-α

peptide comprises at least one glycosyl group comprising a structure having the formula;

15. The IFN-α

conjugate according to claim 14, wherein said at least one glycosyl linking group attached to an amino acid of said IFN-α

peptide has a formula selected from;

16. The IFN-α

conjugate according to claim 15, wherein said amino acid residue is an asparagine residue.

17. A method of preparing an IFN-α

conjugate according to claim 1, said method comprising;

(a) contacting a substrate IFN-α

peptide comprising a glycosyl moiety selected from;

18. The method of claim 17, further comprising, prior to step (a), expressing a nucleic acid encoding said substrate IFN-α

peptide in a suitable host.

19. The method of claim 18, wherein said host is selected from a bacterial cell, an insect cell and a mammalian cell.

20. The method of claim 19, wherein said host is an E. coli cell.

21. The method of claim 19, wherein said host is a Spodoptera frugiperda cell.

22. A method of inhibiting viral replication, increasing the lytic potential of NK cells, and modulating MHC molecule expression, said method comprising administering to a mammal an effective amount of the peptide conjugate according to claim 1.

23. A method of treating a disease in a mammal, wherein said disease is a member selected from hairy cell leukemia, malignant melanoma, follicular lymphoma, condylomata acuminata, AIDS-related Kaposi'"'"'s sarcoma, Hepatitis C, Hepatitis B, human papilloma virus infection, Chronic Myeloid Leukemia (CML), chronic phase Philadelphia chromosome (Ph) positive Chronic Myelogenous Leukemia, non-Hodgkin'"'"'s lymphoma (NHL), lymphoma, bladder cancer, and a renal cancer, said method comprising administering to said mammal an effective amount of a peptide conjugate according to claim 1.

24. A method of treating a viral infection in a subject in need thereof, said method comprising the step of administering to the subject an amount of a peptide conjugate according to claim 1 effective to treat said viral infection in said subject.

25. A pharmaceutical formulation comprising the IFN-α

conjugate according to claim 1, and a pharmaceutically acceptable carrier.

26. A method of preparing an IFN-α

conjugate according to claim 1, said method comprising;

(a) contacting a substrate IFN-α

peptide comprising a glycosyl moiety selected from;

27. The method of claim 26, further comprising, prior to step (a), expressing a nucleic acid encoding the substrate IFN-α

peptide in a suitable host.

28. The method of claim 27, wherein the host is selected from a bacterial cell, an insect cell and a mammalian cell.

29. The method of claim 28, wherein the host is an E. coli cell.

30. The method of claim 28, wherein the host is a Spodoptera frugiperda cell.

31. The method of claim 28, wherein the host is a Chinese hamster ovary (CHO) cell.

13. An IFN-α

conjugate having a formula which is a member selected from the group consisting of;