Human antibodies that bind human TNFalpha

• US 20030092059A1
• Filed: 03/07/2001
• Published: 05/15/2003
• Est. Priority Date: 02/09/1996
• Status: Active Grant

• Current Assignee: AbbVie Biotechnology Ltd. (Puerto Rico) (Abbvie Incorporated)
• Original Assignee: BASF AG (BASF SE)
• Inventors: Hoogenboom, Hendricus R.J.M., Schoenhaut, David, Mcguinness, Brian T., Sakorafas, Paul, White, Michael, Salfeld, Jochen G., Kaymakcalan, Zehra, Labkovsky, Boris, Vaughan, Tristan J., Allen, Deborah J., Roberts, Andrew J., Wilton, Alison J., Mankovich, John A.
• Granted Patent: US 7,223,394 B2
• Time in Patent Office: Days
• US Class Current: 435/7.1
• 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 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/02 Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
• 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 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/16 Central respiratory analeptics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/06 Antigout agents, e.g. antihyperuricemic or uricosuric agents
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• 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 29/02 without antiinflammatory effect
• CPC Class Codes: A61P 3/04 : Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/10 : for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 : Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 : Antibacterial agents
• CPC Class Codes: A61P 31/12 : Antivirals
• CPC Class Codes: A61P 31/18 : for HIV
• CPC Class Codes: A61P 31/20 : for DNA viruses
• CPC Class Codes: A61P 31/22 : for herpes viruses
• CPC Class Codes: A61P 33/06 : Antimalarials
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 35/04 : specific for metastasis
• CPC Class Codes: A61P 37/00 : Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 : Immunomodulators
• CPC Class Codes: A61P 37/04 : Immunostimulants
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 37/08 : Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 39/02 : Antidotes
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 : Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/02 : Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 7/04 : Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 : Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/08 : Vasodilators for multiple indications
• 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 16/241 : Tumor Necrosis Factors
• CPC Class Codes: C07K 2317/21 : from primates, e.g. man
• 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: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
• CPC Class Codes: Y10S 424/81 : involving autoimmunity, allergy, immediate hypersensitivity, delayed hypersensitivity, immunosuppression, immunotolerance, or anergy

Abstract

Human antibodies, preferably recombinant human antibodies, that specifically bind to human tumor necrosis factor a (hTNFα) are disclosed. These antibodies have high affinity for hTNFα (e.g., K_d=10⁻⁸ M or less), a slow off rate for hTNFα dissociation (e.g., K_off=10⁻³ sec⁻¹ or less) and neutralize hTNFα activity in vitro and in vivo. An antibody of the invention can be a full-length antibody or an antigen-binding portion thereof. The antibodies, or antibody portions, of the invention are useful for detecting hTNFα and for inhibiting hTNFα activity, e.g., in a human subject suffering from a disorder in which hTNFα activity is detrimental. Nucleic acids, vectors and host cells for expressing the recombinant human antibodies of the invention, and methods of synthesizing the recombinant human antibodies, are also encompassed by the invention.

Claims

1. An isolated human antibody, or an antigen-binding portion thereof, that dissociates from human TNFα

with a K_d of 1×

10<sup>−

8</sup> M or less and a K_offrate constant of 1×

10<sup>−

3</sup> s<sup>−

1</sup> or less, both determined by surface plasmon resonance, and neutralizes human TNFα

cytotoxicity in a standard in vitro L929 assay with an IC₅₀ of 1×

10<sup>−

7</sup> M or less.

2. The isolated human antibody, or antigen-binding portion thereof, of claim 1, which dissociates from human TNFα

with a K_offrate constant of 5×

10<sup>−

4</sup> s<sup>−

1</sup> or less.

3. The isolated human antibody, or antigen-binding portion thereof, of claim 1, which dissociates from human TNFα

with a K_offrate constant of 1×

10<sup>−

4</sup> s<sup>−

1</sup> or less.

4. The isolated human antibody, or antigen-binding portion thereof, of claim 1, which neutralizes human TNFα

cytotoxicity in a standard in vitro L929 assay with an IC₅₀ of 1×

10<sup>−

8</sup> M or less.

5. The isolated human antibody, or antigen-binding portion thereof, of claim 1, which neutralizes human TNFα

cytotoxicity in a standard in vitro L929 assay with an IC₅₀ of 1×

10<sup>−

9</sup> M or less.

6. The isolated human antibody, or antigen-binding portion thereof, of claim 1, which neutralizes human TNFα

cytotoxicity in a standard in vitro L929 assay with an IC₅₀ of 5×

10<sup>−

10</sup> M or less.

7. The isolated human antibody, or antigen-binding portion thereof, of claim 1, which is a recombinant antibody, or antigen-binding portion thereof.

8. The isolated human antibody, or antigen-binding portion thereof, of claim 1, which inhibits human TNFα

-induced expression of ELAM-1 on human umbilical vein endothelial cells.

45. A pharmaceutical composition comprising the antibody, or antigen-binding portion thereof, of any of claims 1-24, and a pharmaceutically acceptable carrier.

46. The pharmaceutical composition of claim 45, which further comprises at least one additional therapeutic agent for treating a disorder in which TNFα

activity is detrimental.

47. A method for inhibiting human TNFα

activity comprising contacting human TNFα

with the antibody, or antigen-binding portion thereof, of any of claims 1-24 such that human TNFα

activity is inhibited.

48. A method for inhibiting human TNFα

activity in a human subject suffering from a disorder in which TNFα

activity is detrimental, comprising administering to the human subject the antibody, or antigen-binding portion thereof, of any of claims 1-24 such that human TNFα

activity in the human subject is inhibited.

49. The method of claim 48, wherein the disorder is sepsis.

50. The method of claim 49, wherein the antibody is administered to the human subject together with the cytokine interleukin-6 (IL-6) or is administered to a human subject with a serum or plasma concentration of IL-6 above 500 μ

g/ml.

51. The method of claim 48, wherein the disorder is an autoimmune disease.

52. The method of claim 51, wherein the autoimmune disease is selected from the group consisting of rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis and gouty arthritis.

53. The method of claim 51, wherein the autoimmune disease is selected from the group consisting of an allergy, multiple sclerosis, autoimmune diabetes, autoimmune uveitis and nephrotic syndrome.

54. The method of claim 48, wherein the disorder is an infectious disease.

55. The method of claim 48, wherein the disorder is transplant rejection or graft-versus-host disease.

56. The method of claim 48, wherein the disorder is a malignancy.

57. The method of claim 48, wherein the disorder is a pulmonary disorder.

58. The method of claim 48, wherein the disorder is an intestinal disorder.

59. The method of claim 48, wherein the disorder is a cardiac disorder.

60. The method of claim 48, wherein the disorder is selected from the group consisting of inflammatory bone disorders, bone resorption disease, alcoholic hepatitis, viral hepatitis, fulminant hepatitis, coagulation disturbances, burns, reperfusion injury, keloid formation, scar tissue formation, pyrexia, periodontal disease, obesity and radiation toxicity.

61. Use of the antibody, or antigen-binding portion thereof, of any of claims 1-24 in the manufacture of a medicament for the treatment of a a disorder in which TNFα

activity is detrimental.

62. The use of claim 61, wherein the disorder is sepsis.

63. The use of claim 62, wherein the antibody is administered to the human subject together with the cytokine interleukin-6 (IL-6) or is administered to a human subject with a serum or plasma concentration of IL-6 above 500 μ

g/ml.

64. The use of claim 61, wherein the disorder is an autoimmune disease.

65. The use of claim 64, wherein the autoimmune disease is selected from the group consisting of rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis and gouty arthritis.

66. The use of claim 64, wherein the autoimmune disease is selected from the group consisting of an allergy, multiple sclerosis, autoimmune diabetes, autoimmune uveitis and nephrotic syndrome.

67. The use of claim 61, wherein the disorder is an infectious disease.

68. The use of claim 61, wherein the disorder is transplant rejection or graft-versus-host disease.

69. The use of claim 61, wherein the disorder is a malignancy.

70. The use of claim 61, wherein the disorder is a pulmonary disorder.

71. The use of claim 61, wherein the disorder is an intestinal disorder.

72. The use of claim 61, wherein the disorder is a cardiac disorder.

73. The use of claim 61, wherein the disorder is selected from the group consisting of inflammatory bone disorders, bone resorption disease, alcoholic hepatitis, viral hepatitis, fulminant hepatitis, coagulation disturbances, burns, reperfusion injury, keloid formation, scar tissue formation, pyrexia, periodontal disease, obesity and radiation toxicity.

9. An isolated human antibody, or antigen-binding portion thereof, with the following characteristics:

a) dissociates from human TNFα

with a K_offrate constant of 1×

10<sup>−

3</sup> s<sup>−

1</sup> or less, as determined by surface plasmon resonance;

b) has a light chain CDR3 domain comprising the amino acid sequence of SEQ ID NO;

3, or modified from SEQ ID NO;

3 by a single alanine substitution at position 1, 4, 5, 7 or 8 or by one to five conservative amino acid substitutions at positions 1, 3, 4, 6, 7, 8 and/or 9;

c) has a heavy chain CDR3 domain comprising the amino acid sequence of SEQ ID NO;

4, or modified from SEQ ID NO;

4 by a single alanine substitution at position 2, 3, 4, 5, 6, 8, 9, 10 or 11 or by one to five conservative amino acid substitutions at positions 2, 3, 4, 5, 6, 8, 9, 10, 11 and/or 12.

10. The isolated human antibody of claim 9, or an antigen-binding portion thereof, which dissociates from human TNFα

with a K_offrate constant of 5×

10<sup>−

4</sup> s<sup>−

1</sup> or less.

11. The isolated human antibody of claim 9, or an antigen-binding portion thereof, which dissociates from human TNFα

with a K_offrate constant of 1×

10<sup>−

4</sup> s<sup>−

1</sup> or less.

12. An isolated human antibody, or an antigen-binding portion thereof, with a light chain variable region (LCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO:

3, or modified from SEQ ID NO;

3 by a single alanine substitution at position 1, 4, 5, 7 or 8, and with a heavy chain variable region (HCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO;

4, or modified from SEQ ID NO;

4 by a single alanine substitution at position 2, 3, 4, 5, 6, 8, 9, 10 or 11.

13. The isolated human antibody, or an antigen-binding portion thereof, of claim 12, wherein the LCVR further has a CDR2 domain comprising the amino acid sequence of SEQ ID NO:

5 and the HCVR further has a CDR2 domain comprising the amino acid sequence of SEQ ID NO;

6.

14. The isolated human antibody, or an antigen-binding portion thereof, of claim 13, wherein the LCVR further has CDR1 domain comprising the amino acid sequence of SEQ ID NO:

7 and the HCVR has a CDR1 domain comprising the amino acid sequence of SEQ ID NO;

8.

15. An isolated human antibody, or an antigen binding portion thereof, with a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

1 and a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO;

2.

16. The isolated human antibody of claim 15, which has an IgG1 heavy chain constant region.

17. The isolated human antibody of claim 15, which has an IgG4 heavy chain constant region.

18. The isolated human antibody of claim 15, which is a Fab fragment.

19. The isolated human antibody of claim 15, which is a single chain Fv fragment.

20. An isolated human antibody, or an antigen-binding portions thereof, with a light chain variable region (LCVR) having a CDR3 domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:

3, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

16, SEQ ID NO;

17, SEQ ID NO;

18, SEQ ID NO;

19, SEQ ID NO;

20, SEQ ID NO;

21, SEQ ID NO;

22, SEQ ID NO;

23, SEQ ID NO;

24, SEQ ID NO;

25, SEQ ID NO;

26 or with a heavy chain variable region (HCVR) having a CDR3 domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

27, SEQ ID NO;

28, SEQ ID NO;

29, SEQ ID NO;

30, SEQ ID NO;

31, SEQ ID NO;

32, SEQ ID NO;

33 and SEQ ID NO;

34.

21. A recombinant human antibody, or antigen-binding portion thereof, that neutralizes the activity of human TNFα

but not human TNFβ

.

22. The recombinant human antibody, or antigen-binding portion thereof, of claim 21, which also neutralizes the activity of chimpanzee TNFα

and at least one additional primate TNFα

selected from the group consisting of baboon TNFα

, marmoset TNFα

, cynomolgus TNFα and

rhesus TNFα

.

23. The recombinant human antibody, or an antigen-binding portion thereof, of claim 22, which also neutralizes the activity of canine TNFα

.

24. The recombinant human antibody, or an antigen-binding portion thereof, of claim 22, which also neutralizes the activity of pig TNFα

.

25. An isolated nucleic acid encoding a light chain CDR3 domain comprising the amino acid sequence of SEQ ID NO:

3, or modified from SEQ ID NO;

3 by a single alanine substitution at position 1, 4, 5, 7 or 8, or by one to five conservative amino acid substitutions at positions 1, 3, 4, 6, 7, 8 and/or 9.

26. The isolated nucleic acid of claim 25, which encodes an antibody light chain variable region (LCVR).

27. The isolated nucleic acid of claim 26, wherein the CDR2 domain of the antibody LCVR comprises the amino acid sequence of SEQ ID NO:

5.

28. The isolated nucleic acid of claim 27, wherein the CDR1 domain of the antibody LCVR comprises the amino acid sequence of SEQ ID NO:

7.

29. An isolated nucleic acid encoding a heavy chain CDR3 domain comprising the amino acid sequence of SEQ ID NO:

4, or modified from SEQ ID NO;

4 by a single alanine substitution at position 2, 3, 4, 5, 6, 8, 9, 10 or 11, or by one to five conservative amino acid substitutions at positions 2, 3, 4, 5, 6, 8, 9, 10, 11 and/or 12.

30. The isolated nucleic acid of claim 29, which encodes an antibody heavy chain variable region (HCVR).

31. The isolated nucleic acid of claim 30, wherein the CDR2 domain of the antibody HCVR comprises the amino acid sequence of SEQ ID NO:

6.

32. The isolated nucleic acid of claim 31, wherein the CDR1 domain of the antibody HCVR comprises the amino acid sequence of SEQ ID NO:

8.

33. An isolated nucleic acid encoding a CDR3 domain comprising an amino acid sequence selected from the group consisting of:

SEQ ID NO;

3, SEQ ID NO 4, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

16, SEQ ID NO;

17, SEQ ID NO. 18, SEQ ID NO;

19, SEQ ID NO;

20, SEQ ID NO;

21, SEQ ID NO;

22, SEQ ID NO;

23, SEQ ID NO;

24, SEQ ID NO;

25, SEQ ID NO;

26, SEQ ID NO;

27, SEQ ID NO;

28, SEQ ID NO;

29, SEQ ID NO;

30, SEQ ID NO;

31, SEQ ID NO;

32, SEQ ID NO;

33 and SEQ ID NO;

34.

34. An isolated nucleic acid encoding an antibody light chain variable region comprising the amino acid sequence of SEQ ID NO:

1.

35. The isolated nucleic acid of claim 34, which encodes the antibody light chain variable region and an antibody light chain constant region.

36. The isolated nucleic acid of claim 35, which is in a recombinant expression vector.

37. An isolated nucleic acid encoding an antibody heavy chain variable region comprising the amino acid sequence of SEQ ID NO:

2.

38. The isolated nucleic acid of claim 37, which encodes the antibody heavy chain variable region and an antibody heavy chain constant region.

39. The isolated nucleic acid of claim 38, wherein the antibody heavy chain constant region is an IgG1 constant region.

40. The isolated nucleic acid of claim 38, wherein the antibody heavy chain constant region is an IgG4 constant region.

41. The isolated nucleic acid of claim 38, which is in a recombinant expression vector.

42. A recombinant expression vector encoding

a) an antibody light chain having a variable region comprising the amino acid sequence of SEQ ID NO:

1; and

b) an antibody heavy chain having a variable region comprising the amino acid sequence of SEQ ID NO;

2.

43. A host cell into which the recombinant expression vector of claim 42 has been introduced.

44. A method of synthesizing a human antibody that binds human TNFα

, comprising culturing the host cell of claim 43 in a culture medium until a human antibody that binds human TNFα

is synthesized by the cell.