Glycopegylation methods and proteins/peptides produced by the methods

• US 8,853,161 B2
• Filed: 09/27/2011
• Issued: 10/07/2014
• Est. Priority Date: 04/09/2003
• Status: Expired due to Term

• Current Assignee: Novo Nordisk A/S
• Original Assignee: Novo Nordisk A/S
• Inventors: DeFrees, Shawn, Zopf, David A., Bayer, Robert J., Hakes, David James, Bowe, Caryn, Chen, Xi
• Primary Examiner(s): HEARD, THOMAS SWEENEY
• Application Number:
  US13/246,512
• Publication Number:
  US 20120220517A1
• Time in Patent Office: 1,106 Days
• Field of Search: None
• US Class Current: 514/13.5
• CPC Class Codes: A61K 38/1793 for cytokines; for lymphokines; for interferons
• CPC Class Codes: A61K 38/1816 Erythropoietin [EPO]
• CPC Class Codes: A61K 38/185 Nerve growth factor [NGF]; Brain derived neurotrophic factor [BDNF]; Ciliary neurotrophic factor [CNTF]; Glial derived neurotrophic factor [GDNF]; Neurotrophins, e.g. NT-3
• CPC Class Codes: A61K 38/193 Colony stimulating factors [CSF]
• CPC Class Codes: A61K 38/212 IFN-alpha
• CPC Class Codes: A61K 38/215 IFN-beta
• CPC Class Codes: A61K 38/24 Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
• CPC Class Codes: A61K 38/36 Blood coagulation or fibrinolysis factors
• CPC Class Codes: A61K 38/40 Transferrins, e.g. lactoferrins, ovotransferrins
• CPC Class Codes: A61K 38/465 acting on ester bonds (3.1), e.g. lipases, ribonucleases
• CPC Class Codes: A61K 38/47 acting on glycosyl compounds (3.2), e.g. cellulases, lactases
• CPC Class Codes: A61K 38/482 Serine endopeptidases (3.4.21)
• 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 38/57 from animals; from humans A61K38/553, A61K38/556 take precedence
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• 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: A61K 47/61 the organic macromolecular compound being a polysaccharide or a derivative thereof
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 29/00 : Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 : for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/04 : Antibacterial agents
• CPC Class Codes: A61P 31/10 : Antimycotics
• CPC Class Codes: A61P 31/12 : Antivirals
• CPC Class Codes: A61P 31/14 : for RNA viruses
• CPC Class Codes: A61P 31/16 : for influenza or rhinoviruses
• CPC Class Codes: A61P 31/18 : for HIV
• CPC Class Codes: A61P 31/20 : for DNA viruses
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 37/04 : Immunostimulants
• CPC Class Codes: A61P 37/08 : Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/10 : of the posterior pituitary hormones, e.g. oxytocin, ADH
• 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: A61P 7/06 : Antianaemics
• CPC Class Codes: C07K 1/1077 : by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids
• CPC Class Codes: C07K 14/00 : Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
• CPC Class Codes: C07K 16/241 : Tumor Necrosis Factors
• CPC Class Codes: C07K 2317/24 : containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/40 : characterized by post-translational modification
• CPC Class Codes: C07K 2317/41 : Glycosylation, sialylation, or fucosylation
• CPC Class Codes: C07K 2317/52 : Constant or Fc region; Isotype
• CPC Class Codes: C07K 5/0215 : containing natural amino acids, forming a peptide bond via their side chain functional group, e.g. epsilon-Lys, gamma-Glu
• CPC Class Codes: C07K 5/1013 : the side chain containing O or S as heteroatoms, e.g. Cys, Ser
• CPC Class Codes: C07K 5/1016 : and aromatic or cycloaliphatic
• CPC Class Codes: C07K 9/00 : Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof
• CPC Class Codes: C07K 9/008 : directly attached to a hetero atom of the saccharide radical, e.g. actaplanin, avoparcin, ristomycin, vancomycin
• CPC Class Codes: C12N 9/6424 : Serine endopeptidases (3.4.21)
• CPC Class Codes: C12N 9/6437 : Coagulation factor VIIa (3.4.21.21)
• CPC Class Codes: C12N 9/644 : Coagulation factor IXa (3.4.21.22)
• CPC Class Codes: C12N 9/6459 : t-plasminogen activator (3.4.21.68), i.e. tPA
• CPC Class Codes: C12N 9/96 : Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
• CPC Class Codes: C12P 21/005 : Glycopeptides, glycoproteins
• CPC Class Codes: C12Y 301/27005 : Pancreatic ribonuclease (3.1.27.5)
• CPC Class Codes: C12Y 302/01045 : Glucosylceramidase (3.2.1.45), i.e. beta-glucocerebrosidase
• CPC Class Codes: C12Y 304/21021 : Coagulation factor VIIa (3.4.21.21)
• CPC Class Codes: C12Y 304/21022 : Coagulation factor IXa (3.4.21.22)
• CPC Class Codes: C12Y 304/21068 : Tissue plasminogen activator (3.4.21.68), i.e. tPA

Abstract

The invention includes methods and compositions for remodeling a peptide molecule, including the addition or deletion of one or more glycosyl groups to a peptide, and/or the addition of a modifying group to a peptide.

Claims

1. A covalent conjugate between Factor IX and a water-soluble polymer selected from the group consisting of poly(ethers), poly(amines), and poly(carboxylic acids), wherein said water-soluble polymer is covalently attached to Factor IX through an intact glycosyl linking group.

2. The covalent conjugate according to claim 1, wherein said water-soluble polymer is a poly(ether).

3. The covalent conjugate according to claim 2, wherein said poly(ether) is a poly(alkylene oxide).

4. The covalent conjugate according to claim 3, wherein said poly(alkylene oxide) is a poly(ethylene glycol).

5. The covalent conjugate according to claim 4, wherein said poly(ethylene glycol) is a member selected from linear poly(ethylene glycol) and branched poly(ethylene glycol).

6. The covalent conjugate according to claim 4, wherein said poly(ethylene glycol) has a degree of polymerisation of from about 1 to about 5,000.

7. The covalent conjugate according to claim 6, wherein said poly(ethylene glycol) has a degree of polymerisation of from about 1 to about 1,000.

8. The covalent conjugate according to claim 1, wherein said intact glycosyl linking group is attached to a member selected from a carbohydrate moiety, an amino acid moiety and combinations thereof.

9. The covalent conjugate according to claim 8, wherein said intact glycosyl linking group is attached to a carbohydrate moiety, wherein the carbohydrate moiety is an O- or N-linked glycan, or combination thereof.

10. The covalent conjugate according to claim 8, wherein said intact glycosyl linking group is attached to a hydroxyl or amino group or a combination thereof of an amino acid moiety.

11. The covalent conjugate according to claim 8, wherein said intact glycosyl linking group comprises sialic acid, galactose, N-acetylglucosamine or N-acetylgalactosamine residue.

12. The covalent conjugate according to claim 1, wherein said water-soluble polymer is attached to a sialic acid residue at the 5- or 9-position of said sialic acid.

13. A pharmaceutical composition comprising the covalent conjugate according to claim 1 and a pharmaceutically acceptable diluent.

14. A cell-free, in vitro method of forming a covalent conjugate between Factor IX and a water-soluble polymer selected from the group consisting of poly(ethers), poly(amines), and poly(carboxylic acids), wherein said water-soluble polymer is covalently attached to Factor IX through an intact glycosyl linking group, said method comprising:

contacting Factor IX with a mixture comprising at least one nucleotide sugar covalently linked to said water soluble polymer and at least one glycosyltransferase for which said nucleotide sugar is a substrate, thereby forming said covalent conjugate of Factor IX.

15. The method according to claim 14, wherein said water-soluble polymer is a poly(ether).

16. The method according to claim 15, wherein said poly(ether) is a poly(ethylene glycol).

17. The method of claim 14, wherein said glycosyltransferase is selected from the group consisting of sialyltransferase, galactosyltransferase, glucosyltransferase, GalNAc transferase, GlcNAc transferase, fucosyltransferase, and mannosyltransferase.