Interleukin-1 alpha ABS and methods of use

• US 20090298096A1
• Filed: 06/01/2009
• Published: 12/03/2009
• Est. Priority Date: 05/30/2008
• Status: Active Grant

• Current Assignee: Janssen Biotech, Inc. (Johnson & Johnson)
• Original Assignee: XBiotech Inc.
• Inventors: Simard, John
• Granted Patent: US 8,034,337 B2
• Time in Patent Office: Days
• US Class Current: 435/7.21
• CPC Class Codes: A61K 2039/55527 Interleukins
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• 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 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/5412 IL-6
• CPC Class Codes: C07K 14/545 IL-1
• CPC Class Codes: C07K 16/24 against cytokines, lymphokines or interferons
• CPC Class Codes: C07K 16/245 IL-1
• CPC Class Codes: C07K 2317/14 Specific host cells or culture conditions, e.g. components, pH or temperature
• 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 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C12N 2330/50 Biochemical production, i.e. in a transformed host cell
• CPC Class Codes: C12N 5/069 Vascular Endothelial cells
• CPC Class Codes: C12N 5/0693 Tumour cells; Cancer cells
• CPC Class Codes: G01N 2333/545 : IL-1
• CPC Class Codes: G01N 33/6863 : Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
• CPC Class Codes: G01N 33/6869 : Interleukin

Abstract

Fully human monoclonal Abs includes (i) an antigen-binding variable region that exhibits very high binding affinity for IL-1α and (ii) a constant region that is effective at both activating the complement system though C1q binding and binding to several different Fc receptors.

Claims

1-22. -22. (canceled)

23. A purified human IgG1 mAb that specifically binds to human IL-1α

, the mAb comprising a heavy chain covalently joined to a light chain.

24. The purified human IgG1 monoclonal Ab of claim 23, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:

9 and the light chain comprises the amino acid sequence of SEQ ID NO;

11.

25. A set of isolated nucleic acids comprising a first nucleic acid encoding the heavy chain of a human IgG1 mAb that specifically binds to IL-1α

, and a second nucleic acid encoding the light chain of the human IgG1 mAb that specifically binds to human IL-1α

.

26. The set of isolated nucleic acids of claim 25, wherein the first nucleic acid encodes the amino acid sequence of SEQ ID NO:

9 and the second nucleic acid encodes the amino acid sequence of SEQ ID NO;

11.

27. The set of nucleic acids of claim 26, wherein the first nucleic acid comprises the nucleotide sequence of SEQ ID NO:

10 and the second nucleic acid comprises the nucleotide sequence of SEQ ID NO;

12.

28. The set of isolated nucleic acids of claim 25, wherein the set of isolated nucleic acids is comprised within at least one expression vector.

29. The set of isolated nucleic acids of claim 26, wherein the set of isolated nucleic acids is comprised within at least one expression vector.

30. The set of isolated nucleic acids of claim 27, wherein the set of isolated nucleic acids is comprised within at least one expression vector.

31. The set of isolated nucleic acids of claim 25, wherein the set of isolated nucleic acids is comprised within an isolated host cell.

32. The set of isolated nucleic acids of claim 26, wherein the set of isolated nucleic acids is comprised within an isolated host cell.

33. The set of isolated nucleic acids of claim 32, wherein the host cell is a mammalian cell.

34. A method of killing a cell expressing human IL-1α

, the method comprising the step of contacting the cell with a purified human IgG1 mAb that specifically binds to human IL-1α

.

35. The method of claim 34, wherein the human IgG1 mAb comprises a heavy chain comprising the amino acid sequence of SEQ ID NO. 9 and a light chain comprising the amino acid sequence of SEQ ID NO:

11.

36. A method of inhibiting migration of a cell through a basement membrane matrix, the method comprising the step of adding a purified mAb that specifically binds to human IL-1α

to a mixture comprising the basement membrane matrix and the cell.

37. The method of claim 36, wherein the purified mAb is a human IgG1 mAb comprising a heavy chain covalently joined to a light chain.

38. The method of claim 37, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:

9 and a light chain comprises the amino acid sequence of SEQ ID NO;

11.

39. A method of inhibiting an IL-1α

-induced increase in expression of ICAM-1 or E-selectin on the surface of a human endothelial cell, the method comprising the step of adding a purified mAb that specifically binds to human IL-1α

to a mixture comprising the endothelial cell and IL-1α

.

40. The method of claim 39, wherein the purified mAb is a human IgG1 mAb comprising a heavy chain covalently joined to a light chain.

41. The method of claim 40, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:

9 and a light chain comprises the amino acid sequence of SEQ ID NO;

11.

42. A method of tracking inflammation in a human subject previously subjected to the steps of:

obtaining from the subject a first sample of peripheral blood mononuclear cells at a first time;

contacting the first sample with a purified mAb that specifically binds to human IL-1α

; and

determining the percent of cells in the first sample that bind the monoclonal Ab, the method comprising the steps of;

(a) obtaining from the subject a second sample of peripheral blood mononuclear cells at a second time;

(b) contacting the second sample with the purified mAb that specifically binds to human IL-1α

;

(c) determining the percent of cells in the second sample that bind the monoclonal Ab; and

(d) comparing the percent of cells in the first sample that bind the mAb to the percent of cells in the second sample that bind the monoclonal Ab.

43. The method of claim 42, wherein the purified mAb is a human IgG1 mAb comprising a heavy chain covalently joined to a light chain.

44. The method of claim 43, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:

9 and a light chain comprises the amino acid sequence of SEQ ID NO;

1.

45. A method comprising the steps of:

(a) enriching a biological sample obtained from a human subject using a filter that separates molecules according to molecular weight into a first fraction comprising intact IgG complexed with IL-1α and

second fraction comprising molecule less than 100 Kda; and

(b) quantifying the amount of IL-1α

in the first fraction.

46. The method of claim 45, comprising the steps of:

(a) enriching a sample of plasma obtained from a human subject using a filter that separates molecules according to molecular weight into a first fraction comprising intact IgG complexed with IL-1α and

second fraction comprising molecules less than 100 Kda;

(b) adding the first fraction to a substrate comprising immobilized anti-human IgG Abs under conditions that allow IgG in the first fraction to specifically bind the anti-human IgG Abs immobilized on the substrate;

(c) washing the substrate to remove material in the first fraction that does not specifically bind the immobilized anti-human IgG Abs;

(d) contacting the substrate washed in step (c) with an Ab that specifically binds human IL-1α

under conditions that allows the Ab that specifically binds human IL-1α

to specifically bind any human IL-1α

bound to the substrate;

(e) washing the substrate to remove any of the Ab that specifically binds human IL-1α

that is not bound to the substrate; and

(f) quantifying the amount of Ab that specifically binds human IL-1α

remaining bound to the substrate after step (e).