Methods for generating monovalent IgG

• US 8,193,322 B2
• Filed: 10/23/2006
• Issued: 06/05/2012
• Est. Priority Date: 10/21/2005
• Status: Active Grant

• Current Assignee: Amgen Inc.
• Original Assignee: Amgen Inc.
• Inventors: Yan, Wei, Wittekind, Michael, Forte, Carla
• Primary Examiner(s): BRISTOL, LYNN ANNE
• Application Number:
  US11/585,505
• Publication Number:
  US 20070105199A1
• Time in Patent Office: 2,052 Days
• Field of Search: 530/388.2, 530/391.1
• US Class Current: 530/388.22
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 15/08 for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• 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 37/02 Immunomodulators
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/00 Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 5/14 of the thyroid hormones, e.g. T3, T4
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/2878 against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/30 : Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Abstract

The present invention relates to monovalent antibody, methods of making thereof and therapeutic uses thereof. In particular, the present invention provides a heterodimeric polypeptide comprising an immunoglobulin heavy chain and a fusion protein comprising an immunoglobulin light chain and an Fc molecule.

Claims

1. A heterodimeric polypeptide comprising

(a) a human or humanized IgG1, IgG2, or IgG4 heavy chain from an antibody that binds specifically to an antigen and that comprises an Fe region, wherein the heavy chain comprises a heavy chain variable (V_H) region, followed by a first heavy chain constant (C_H1) region, followed by a hinge region, followed by a second heavy chain constant (C_H2) region, followed by a third heavy chain constant (C_H3) region, and

(b) a fusion protein, wherein the fusion protein comprises a light chain from the antibody followed by a human IgG1, IgG2, or IgG4 Fe molecule, wherein the light chain comprises a light chain variable (V_L) region followed by a light chain constant (C_L) region and wherein the human IgG1, IgG2, or IgG4 Fe molecule comprises a hinge region followed by a C_H 2 region followed by a C_H 3 region, wherein

(i) the first five amino acids of the hinge region of the fusion protein are deleted, or

(ii) the hinge region of the fusion protein is not shortened or lengthened,

wherein the heavy chain and the fusion protein are two polypeptide chains that are oriented identically with respect to their N- and C-termini and associate to form the heterodimeric polypeptide,

wherein the heterodimeric polypeptide is monovalent, and

wherein the heterodimeric polypeptide is capable of specifically binding to the antigen.

2. The heterodimeric polypeptide of claim 1, wherein the heavy chain and the fusion protein are not mutated in their CH3 regions.

3. The heterodimeric polypeptide of claim 1, wherein the heavy chain and the fusion protein are mutated in their CH3 regions so as to form multimerization domains.

4. The heterodimeric polypeptide of claim 3, wherein the mutations are substitutions.

5. The heterodimeric polypeptide of claim 1, wherein the light chain is a kappa light chain.

6. The hoterodimeric polypeptide of claim 1, wherein the light chain is a lambda light chain.

7. The heterodimeric polypeptide of claim 1, wherein the antibody is a human antibody.

8. The heterodimeric polypeptide of claim 1, wherein the antibody is a humanized antibody.

9. The heterodimeric polypeptide of claim 1, wherein the heavy chain and the fusion protein are covalently linked.

10. The heterodimeric polypeptide of claim 9, wherein the covalent linkage is a disulfide bond.

11. The heterodimeric polypeptide of claim 1, wherein the hinge region of the fusion protein is not shortened or lengthened.

12. The heterodimeric polypeptide of claim 1, wherein the first five amino acids of the binge region of the fusion protein are deleted.

13. The heterodimeric polypeptide of claim 1, wherein the heavy chain further comprises a tagging moiety.

14. The heterodimeric polypeptide of claim 13, wherein the tagging moiety is selected from the group consisting of:

hexa-histidine tag, streptavidin-binding peptide, maltose-binding protein, glutathione S-transferase, mic-tag, and FLAG-tag.

15. The heterodimeric polypeptide of claim 14, wherein the tagging moiety comprises a hexa-histidine tag.

16. The heterodimeric polypeptide according to claim 1, wherein the sequences of the hinge region are naturally occurring sequences.

17. The heterodimeric polypeptide of claim 1, wherein the heavy chain and the lusion protein contain no cysteine residues that are unpaired.

18. The heterodimeric polypeptide of claim 12, wherein the Fe region of the heavy chain and the Fe molecule of the fusion protein are of the IgG1 isotype.

19. The heterodimeric polypeptide of claim 11, wherein the Fe region of the heavy chain and the Fe molecule of the fusion protein are of the IgG2 isotype.

20. The heterodimerie polypeptide of claim 11, wherein the Fe region of the heavy chain and the Fe molecule of the fusion protein are of the IgG4 isotype.

21. The heterodimeric polypeptide of claim 1, wherein the antigen is a receptor which is activated by the oligoimerization of its subunits.

22. The heterodimeric polypeptide of claim 21, which antagonizes the activity of the receptor.

23. The heterodimeric polypeptide of claim 21, wherein the oligomerization forms an oligomer which is at least a dimer.

24. The heterodimeric polypeptide of claim 23, wherein the oligomerizatim forms an oligomer which is at least a trimer.

25. The heterodimeric polypeptide of claim 24, wherein the oligomerization forms an oligomer which is at least a tetramer.

26. The heterodimeric polypeptide of claim 21, wherein the receptor is a member of a family selected from the group consisting of:

TNF/TNF receptor superfamily, cytokine receptors, receptor tyrosine kinases, G-protein coupled receptors (GPCRs), Fe receptors (FeRs), AT1 receptors, tissue factors, and integrins.

27. The heterodimeric polypeptide of claim 26, wherein the receptor is a member of the TNF/TNF receptor superfamily.

28. The heterodimeric of claim 27, wherein the member is selected from the group consisting of:

TNFR1, TNFR2, NGFR, Troy, EDAR, XEDAR, CD40, DeR3, Fas, OX40, AITR, CD30,HveA, 4-1BB, DR3, CD27, LTβ

R, RANK, TWEAK receptor, TACI, BCMA, DR6, OPG, DR4, DR5, DeR1 and DeR2.

29. The heterodimeric polypeptide of claim 26, wherein the receptor is a receptor tyrosine kinase.

30. The heterodimeric polypeptide of claim 29, wherein the receptor tyrosine kinase is selected from the group consisting of:

growth hormone receptor, erythropoietin receptor, VEGF receptor, FGF receptor, PFGF receptor and EGF receptor.

31. The heterodimeric polypeptide of claim 30, wherein the receptor tyrosine kinase is an EGF receptor selected from the group consisting of ErbB1, ErbB2, ErbB3 and ErbB4.

32. The heterodimeric polypeptide of claim 26, wherein the receptor is an interleukin receptor.

33. The heterodimeric polypeptide of claim 32, wherein the interleukin receptor is a receptor of an interleukin selected from the group consisting of:

IL-1, IL-2, IL-4, IL-15, IL7, TSLP, LIF, IL-13, IL-23 and IL-31.

34. The heterodimeric polypeptide of claim 21, which does not agonize the activity of the receptor.

35. The heterodimeric polypeptide of claim 1, wherein the heavy chain comprises CDRs having the amino acid sequences of SEQ ID NOS:

11, 12 and 13, and the light chain comprises CDRs having the amino acid sequences of SEQ ID NOS;

18, 19 and 20 and wherein the heterodimeric polypeptide is capable of binding to and inhibiting the activity of TNFR1.

36. The heterodimeric polypeptide of claim 1, wherein the heavy chain and the fusion protein form a symmetrical structure beyond the fusion juncture of the fusion protein.

37. The heterudimeric polypeptide of claim 3, wherein the heavy chain and the fusion protein are mutated at one or more interface amino acid residues in their CH3 regions.

38. The heterodimeric polypeptide of claim 3, wherein the heavy chain and the fusion protein are mutated at one or more contact amino acid residues in their CH3 regions.

39. A. composition comprising the heterodimeric polypeptide of claim 1 and a pharmaceutically acceptable carrier.

40. An isolated heterodimeric polypeptide comprising a first polypeptide comprising the amino acid sequence set forth in SEQ ID NO:

9 and a second polypeptide comprising the amino acid sequence set forth in SEQ ID NO;

10.