Glycopegylated erythropoietin

• US 8,916,360 B2
• Filed: 06/10/2011
• Issued: 12/23/2014
• Est. Priority Date: 11/24/2003
• Status: Expired due to Fees

• Current Assignee: Novo Nordisk A/S
• Original Assignee: Novo Nordisk A/S
• Inventors: DeFrees, Shawn, Zopf, David A., Bowe, Caryn Lang
• Primary Examiner(s): GOON, SCARLETT Y
• Application Number:
  US13/157,575
• Publication Number:
  US 20120107867A1
• Time in Patent Office: 1,292 Days
• Field of Search: 435/68.1, 514/7.7, 514/20.9
• US Class Current: 435/68.1
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/22 Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin A61K38/33, e.g. corticotropin A61K38/35
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: C12P 21/005 Glycopeptides, glycoproteins

Abstract

The present invention provides conjugates between erythropoietin 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 cell-free, in vitro method of forming a covalent conjugate between an erythropoietin peptide and poly(ethylene glycol), which method comprises:

(a) providing an erythropoietin peptide,

(b) providing a modified sugar donor comprising an intact glycosyl linking group covalently linked to poly(ethylene glycol), wherein the intact glycosyl linking group has a structure according to the formula;

2. The method of claim 1, wherein the poly(ethylene glycol) has a molecular weight distribution that is essentially homodisperse.

3. The method of claim 1, wherein the intact glycosyl linking group has a structure according to the formula:

4. The method of claim 1, wherein the intact glycosyl linking group is covalently bound to a member selected from the group consisting of (a) an amino acid residue of the peptide which is a member selected from Ser, Thr and Asn;

(b) a glycosyl residue covalently bound to an amino acid residue of the peptide which is a member selected from Ser, Thr and Asn; and

(c) combinations thereof.

5. The method of claim 3, wherein a is 0 and b is 0.

6. The method of claim 3, wherein a is 0 and b is 1.

7. The method of claim 3, wherein a is 1 and b is 0.

8. The method of claim 3, wherein a is 1 and b is 1.