Methods of binding TNF-alpha using Anti-TNF-alpha antibody fragment-polymer conjugates

• US 20130084302A1
• Filed: 02/13/2012
• Published: 04/04/2013
• Est. Priority Date: 01/21/1999
• Status: Active Grant

• Current Assignee: Gerardo Zapata, Iphigenia Koumenis, Steven Leong, Vanessa Hsei, Zahra Shahrokh
• Original Assignee: Gerardo Zapata, Iphigenia Koumenis, Steven Leong, Vanessa Hsei, Zahra Shahrokh
• Inventors: Hsei, Vanessa, Koumenis, Iphigenia, Leong, Steven, Shahrokh, Zahra, Zapata, Gerardo
• Granted Patent: US 8,652,468 B2
• Time in Patent Office: Days
• US Class Current: 424/178.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• 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 47/6845 the antibody targeting a cytokine, e.g. growth factors, VEGF, TNF, a lymphokine or an interferon
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• 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 31/04 Antibacterial agents
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/244 Interleukins [IL]
• CPC Class Codes: C07K 17/02 Peptides being immobilised on, or in, an organic carrier
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/53 Hinge
• CPC Class Codes: C07K 2317/54 F(ab')2
• CPC Class Codes: C07K 2317/55 : Fab or Fab'
• CPC Class Codes: C07K 2317/56 : variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 : Complementarity determining region [CDR]
• 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

Abstract

Described are conjugates formed by an antibody fragment covalently attached to a nonproteinaceous polymer, wherein the apparent size of the conjugate is at least about 500 kD. The conjugates exhibit substantially improved half-life, mean residence time, and/or clearance rate in circulation as compared to the underivatized parental antibody fragment. Also described are conjugates directed against human vascular endothelial growth factor (VEGF), human p185 receptor-like tyrosine kinase (HER2), human CD20, human CD18, human CD11a, human IgE, human apoptosis receptor-2 (Apo-2), human tumor necrosis factor-α (TNF-α), human tissue factor (TF), human α₄β₇ integrin, human GPIIb-IIIa integrin, human epidermal growth factor receptor (EGFR), human CD3, and human interleukin-2 receptor α-chain (TAC) for diagnostic and therapeutic applications.

Claims

1-32. -32. (canceled)

33. A method of binding a conjugate to a tumor necrosis factor α

(TNF-α

) polypeptide in a mammal comprising a TNF-α

polypeptide, the method comprising administering to said mammal a conjugate consisting essentially of an antibody fragment covalently modified by one or two nonproteinaceous polymer molecules at a free sulfhydryl group of a cysteine residue within the hinge region of the antibody fragment,

wherein said antibody fragment is engineered to both (1) provide an unpaired cysteine within the hinge region, so as to provide said free sulfhydryl group, and (2) to avoid disulfide bridge formation between said cysteine and an amino acid in the opposite chain of said antibody fragment,

wherein (a) the average actual molecular weight of each nonproteinaceous polymer molecule is at least 20 kD, (b) the conjugate binds the same antigen as the parental molecule that is not covalently modified by one or two nonproteinaceous polymer molecules, wherein the antibody fragment comprises an antigen binding site that binds to a human TNF-α

polypeptide.

34. The method of claim 33 wherein the mammal suffers from Crohn'"'"'s disease.

35. The method of claim 33 wherein the mammal suffers from inflammatory bowel disease.

36. The method of claim 33 wherein the mammal suffers from rheumatoid arthritis.

37. The method of claim 35, wherein the inflammatory bowel disease is ulcerative colitis.

38. The method of claim 33 wherein the mammal suffers from an inflammation caused by neutrophil chemotaxis.

39. The method of claim 33 wherein the mammal suffers from an inflammatory disorder selected from inflammatory skin disease, systemic scleroderma, sclerosis, inflammatory bowel disease, a myocardial ischemic condition, an ischemic reperfusion disorder, stroke, reperfusion after stroke, cerebral edema secondary to stroke, cranial trauma, hypovolemic shock, asphyxia, adult respiratory distress syndrome, acute lung injury, Behcet'"'"'s Disease, dermatomyositis, polymyositis, multiple sclerosis, dermatitis, meningitis, encephalitis, uveitis, osteoarthritis, lupus nephritis, an autoimmune disease, a disease involving leukocyte diapedesis, central nervous system (CNS) inflammatory disorder, multiple organ injury syndrome secondary to septicaemia or trauma, alcoholic hepatitis, asthma, bacterial pneumonia, an antigen-antibody complex mediated disease, sepsis, sarcoidosis, an immunopathologic response to tissue/organ transplantation, and an inflammation of the lung.

40. The method of claim 39, wherein the inflammatory disorder is selected from acute lung injury, adult respiratory distress syndrome, an ischemic reperfusion disorder, hypovolemic shock, asthma, bacterial pneumonia and inflammatory bowel disease.

41. The method of claim 39, wherein the inflammatory disorder is an ischemic reperfusion disorder selected from surgical tissue reperfusion injury, myocardial infarction, cardiac arrest, reperfusion after cardiac surgery, constriction after percutaneous transluminal coronary angioplasty, stroke, reperfusion after stroke, cerebral edema secondary to stroke, and an abdominal aortic aneurysm.

42. The method of claim 39, wherein the inflammatory disorder is an inflammatory skin disease selected from psoriasis and atopic dermatitis.

43. The method of claim 39, wherein the inflammatory disorder is an autoimmune disease selected from rheumatoid arthritis, Sjorgen'"'"'s syndrome, and vasculitis.

44. The method of claim 39, wherein the inflammatory disorder is an inflammation of the lung selected from pleurisy, alveolitis, vasculitis, pneumonia, chronic bronchitis, bronchiectasis, diffuse panbronchiolitis, hypersensitivity pneumonitis, idiopathic pulmonary fibrosis (IPF), and cystic fibrosis.

45. The method of claim 39, wherein the inflammatory disorder is an antigen-antibody complex mediated disease.

46. The method of claim 45, wherein the antigen-antibody complex mediated disease is glomerulonephritis.

47. The method of claim 39, wherein the inflammatory disorder is systemic scleroderma or sclerosis.

48. The method of claim 41, wherein the inflammatory disorder is an ischemic reperfusion disorder selected from surgical tissue reperfusion injury, myocardial ischemia, and acute myocardial infarction.

49. The method of claim 39, where in the inflammatory disorder is reperfusion after stroke.

50. The method of claim 33, wherein said binding affects the activity of said TNF-α

polypeptide in said mammal.

51. The method of claim 50, wherein said mammal suffers from an inflammatory disorder.