DEGRADED AGONIST ANTIBODY

• US 20090311718A1
• Filed: 07/02/2009
• Published: 12/17/2009
• Est. Priority Date: 10/20/2000
• Status: Abandoned Application

• Current Assignee: Chugai Seiyaku Kabushiki Kaisha (Roche Holding AG)
• Original Assignee: Chugai Seiyaku Kabushiki Kaisha (Roche Holding AG)
• Inventors: Tsuchiya, Masayuki, Ohtomo, Toshihiko, Yabuta, Naohiro, Fukushima, Naoshi, Uno, Shinsuke, Tsunod, Hiroyuki
• Application Number:
  US12/497,131
• Publication Number:
  US 20090311718A1
• Time in Patent Office: Days
• US Class Current: 435/7.200
• CPC Class Codes: A61K 2039/505 comprising antibodies
• 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: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• 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 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: C07K 16/24 against cytokines, lymphokines or interferons
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/2866 against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C07K 16/2869 against hormone receptors for antibodies against neuromediator receptors C07K16/286
• CPC Class Codes: C07K 16/3061 Blood cells
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/73 : Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide

Abstract

The invention relates to a modified antibody which contains two or more H chain V regions and two or more L chain V regions of monoclonal antibody and can transduce a signal into cells by crosslinking a cell surface molecule(s) to thereby serve as an agonist. The modified antibody can be used as a signal transduction agonist and, therefore, useful as a preventive and/or remedy for various diseases such as cancer, inflammation, hormone disorders and blood diseases.

Claims

1. A modified antibody comprising two or more H chain V regions and two or more L chain V regions of the same or different monoclonal antibody and showing an agonist action by crosslinking a cell surface molecule(s) or intracellular molecule(s).

3. The modified antibody of claim 1 or 2, wherein the H chain V region and the L chain V region are connected through a linker.

4. The modified antibody of claim 3, wherein the linker is a peptide linker comprising at least one amino acid.

5. The modified antibody of any one of claims 1 to 4, wherein the modified monoclonal antibody is a multimer of single chain Fv comprising an H chain V region and an L chain V region.

6. The modified antibody of claim 5, wherein the modified antibody is composed of tetramer, trimer or dimer of single chain Fv.

7. The modified antibody of claim 6, wherein the modified antibody is composed of dimer of single chain Fv.

8. The modified antibody of any one of claims 5 to 7, wherein the H chain V region and the L chain V region existing in the same chain are not associated to form an antigen-binding site.

9. The modified antibody of any one of claims 1 to 4, wherein the modified antibody is a single chain polypeptide comprising two or more H chain V regions and two or more L chain V regions.

10. The modified antibody of claim 9, wherein the modified antibody is a single chain polypeptide comprising two H chain V regions and two L chain V regions.

11. The modified antibody of any one of claims 1 to 10, wherein the modified antibody further comprises an amino acid sequence(s) for peptide purification.

12. The modified antibody of any one of claims 1 to 11, wherein the modified antibody has been purified.

13. The modified antibody of any one of claims 1 to 12, wherein H chain V region and/or L chain V region is H chain V region and/or L chain V region derived from a human antibody.

14. The modified antibody of any one of claims 1 to 13, wherein H chain V region and/or L chain V region is humanized H chain V region and/or L chain V region.

15. The modified antibody of any one of claims 1 to 14, wherein the cell surface molecule or intracellular molecule is a hormone receptor, a cytokine receptor, tyrosine kinase receptor or intranuclear receptor.

16. The modified antibody of any one of claims 1 to 15, wherein the cell surface molecule or intracellular molecule is erythropoietin (EPO) receptor, thrombopoietin (TPO) receptor, granulocyte colony stimulating factor (G-CSF) receptor, macrophage colony stimulating factor (M-CSF) receptor, granular macrophage colony stimulating factor (GM-CSF) receptor, tumor necrosis factor (TNF) receptor, interleukin-1 (IL-1) receptor, interleukin-2 (IL-2) receptor, interleukin-3 (IL-3) receptor, interleukin-4 (IL-4) receptor, interleukin-5 (IL-5) receptor, interleukin-6 (IL-6) receptor, interleukin-7 (IL-7) receptor, interleukin-9 (IL-9) receptor, interleukin-10 (IL-10) receptor, interleukin-11 (IL-11) receptor, interleukin-12 (IL-12) receptor, interleukin-13 (IL-13) receptor, interleukin-15 (IL-15) receptor, interferon-alpha (IFN-alpha) receptor, interferon-beta (IFN-beta) receptor, interferon-gamma (IFN-gamma) receptor, growth hormone (GH) receptor, insulin receptor, blood stem cell proliferation factor (SCF) receptor, vascular endothelial growth factor (VEGF) receptor, epidermal cell growth factor (EGF) receptor, nerve growth factor (NGF) receptor, fibroblast growth factor (FGF) receptor, platelet-derived growth factor (PDGF) receptor, transforming growth factor-beta (TGF-beta) receptor, leukocyte migration inhibitory factor (LIF) receptor, ciliary neurotrophic factor (CNTF) receptor, oncostatin M (OSM) receptor, Notch family receptor, E2F, E2F/DP1 or TAK1/TAB1.

17. The modified antibody of any one of claims 1 to 16, wherein the agonist action is apoptosis induction, cell proliferation induction, cell differentiation induction, cell division induction or cell cycle regulation action.

18. The modified antibody of any one of claims 1 to 17, wherein the modified antibody is mono-specific modified antibody.

19. The modified antibody of any one of claims 1 to 17, wherein the modified antibody is multi-specific modified antibody.

20. The modified antibody of claim 19, wherein the modified antibody is bi-specific modified antibody.

21. The monoclonal antibody of claim 20, wherein the L chain V region and the H chain V region are from the same monoclonal antibody.

22. The monoclonal antibody of any one of claims 1 to 21 which shows an equivalent or better agonist action (ED50) compared with the parent monoclonal antibody.

23. The monoclonal antibody of claim 22 which shows at least 2-fold agonist action (ED50) compared with the parent monoclonal antibody.

24. The monoclonal antibody of claim 23 which shows at least 10-fold agonist action (ED50) compared with the parent monoclonal antibody.

25. The monoclonal antibody of any one of claims 1 to 21 which is derived from a parent antibody having substantially no agonist action.

29. The modified antibody or compound of any one of claims 1 to 28 which has substantially no intercellular adhesion action.

30. The modified antibody or compound of any one of claims 1 to 28 which has intercellular adhesion action (ED50) not more than 1/10 compared with the parent antibody.

31. A DNA which encodes the modified antibody or compound of any one of claims 1 to 28.

32. An animal cell which produces the modified antibody or compound of any one of claims 1 to 28.

33. A microorganism which produces the modified antibody or compound of any one of claims 1 to 28.

34. Use of the modified antibody or compound of any one of claims 1 to 28 as an agonist.

38. A method of causing agonist action to cells by crosslinking a cell surface molecule(s) or intracellular molecule(s) using the modified antibody or compound of any one of claims 1 to 28.

39. The method of claim 38 wherein the agonist action is apoptosis induction, cell proliferation induction, cell differentiation induction, cell division induction or cell cycle regulation action.

40. A medicine comprising as active ingredient the modified antibody or compound of any one of claims 1 to 29.

41. Use of the modified antibody or compound of any one of claims 1 to 29 as medicine.

44. A method of producing a modified antibody comprising two or more H chain V regions and two or more L chain V regions of monoclonal antibody and showing an agonist action by crosslinking a cell surface molecule(s) or intracellular molecule(s) which comprises the steps

(1) to culture animal cells of claim 32 or microorganisms of claim 33 to produce the modified antibody and

(2) to purify said monoclonal antibody.

2. The modified antibody comprising two or more H chain V regions and two or more L chain V regions of monoclonal antibody and showing an agonist action by crosslinking a cell surface molecule(s).

26. A compound comprising two or more H chain V regions and two or more L chain V regions of monoclonal antibody and showing an equivalent or better agonist action (ED50) compared with a natural ligand that binds to a cell surface molecule(s) or intracellular molecule(s).

27. The compound of claim 26 which shows at least 2-fold agonist action (ED50) compared with a natural ligand that binds to a cell surface molecule(s) or intracellular molecule(s).

28. The compound of claim 27 which shows at least 10-fold agonist action (ED50) compared with a natural ligand that binds to a cell surface molecule(s) or intracellular molecule(s).

35. A method of inducing an agonist action to cells which comprises administering the first ligand and the second ligand that bind to a cell surface molecule(s) or intracellular molecule(s) and administering a substance that binds to the first and the second ligands and crosslinks the first and the second ligands.

36. The method of claim 35 wherein the first and the second ligands are the same or different single chain Fv monomers.

37. The method of claim 35 or 36 wherein the substance that crosslinks the ligands is an antibody, an antibody fragment or a bivalent modified antibody.

42. A method of screening a modified antibody comprising two or more H chain V regions and two or more L chain V regions of antibody and showing an agonist action by crosslinking a cell surface molecule(s) or intracellular molecule(s) which comprises the steps

(1) to produce a modified antibody comprising two or more H chain V regions and two or more L chain V regions of antibody and binding specifically to a cell surface molecule(s) or intracellular molecule(s),

(2) to subject cells expressing said cell surface molecule(s) or intracellular molecule(s) to react with the modified antibody and

(3) to measure the agonist action in the cells caused by crosslinking said cell surface molecule(s) or intracellular molecule(s).

43. A method of measuring an agonist action of a modified antibody comprising two or more H chain V regions and two or more L chain V regions of antibody and showing an agonist action by crosslinking a cell surface molecule(s) or intracellular molecule(s) which comprises the steps

(1) to produce a modified antibody comprising two or more H chain V regions and two or more L chain V regions of antibody and binding specifically to a cell surface molecule(s) or intracellular molecule(s),

(2) to subject cells expressing said cell surface molecule(s) or intracellular molecule(s) to react with the modified antibody and

(3) to measure the agonist action in the cells caused by crosslinking said cell surface molecule(s) or intracellular molecule(s).