Human antibodies that bind human TNF.alpha.

• US 6,090,382 A
• Filed: 02/09/1996
• Issued: 07/18/2000
• Est. Priority Date: 02/09/1996
• Status: Expired due to Term

• Current Assignee: AbbVie Biotechnology Ltd. (Puerto Rico) (Abbvie Incorporated)
• Original Assignee: BASF AG (BASF SE)
• Inventors: McGuinness, Brian T., Sakorafas, Paul, White, Michael, Salfeld, Jochen G., Kaymakcalan, Zehra, Labkovsky, Boris, Hoogenboom, Hendricus R. J. M., Vaughan, Tristan J., Allen, Deborah J., Roberts, Andrew J., Wilton, Alison J., Mankovich, John A., Schoenhaut, David
• Primary Examiner(s): Saunders, David
• Application Number:
  US08/599,226
• Time in Patent Office: 1,621 Days
• Field of Search: 424/130.1, 424/133.1, 424/135.1, 424/142.1, 424/145.1, 350/387.3, 350/388.23, 350/388.24, 350/388.15
• US Class Current: 424/133.1
• 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 1/04 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 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 3/10 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 31/22 : 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 37/06 : 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 7/00 : Drugs for disorders of the blood or the extracellular fluid
• 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 9/10 : 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: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• 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 424/80 : Antibody or fragment thereof whose amino acid sequence is disclosed in whole or in part
• CPC Class Codes: Y10S 424/81 : involving autoimmunity, allergy, immediate hypersensitivity, delayed hypersensitivity, immunosuppression, immunotolerance, or anergy
• CPC Class Codes: Y10S 530/868 : involving autoimmunity, allergy, immediate hypersensitivity, delayed hypersensitivity, immunosuppression, or immunotolerance

Abstract

Human antibodies, preferably recombinant human antibodies, that specifically bind to human tumor necrosis factor α (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^-8 M or less and a K_off rate constant of 1×

10^-3 s^-1 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^-7 M or less.

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

with a K_off rate constant of 5×

10^-4 s^-1 or less.

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

with a K_off rate constant of 1×

10^-4 s^-1 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^-8 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^-9 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 1×

10^-10 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.

24. A pharmaceutical composition comprising the antibody, or antigen-binding portion thereof, of any one of claims 1-23, and a pharmaceutically acceptable carrier.

25. The pharmaceutical composition of claim 24, which further comprises at least one additional therapeutic agent.

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

a) dissociates from human TNFα

with a K_off rate constant of 1×

10^-3 s^-1 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_off rate constant of 5×

10^-4 s^-1 or less.

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

with a K_off rate constant of 1×

10^-4 s^-1 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. An isolated human antibody, or antigen-binding portion thereof, that neutralizes the activity of human TNFα

, chimpanzee TNFα and

at least one additional primate TNFα

selected from the group consisting of baboon TNFα

, marmoset TNFα

, cynomolgus TNFα and

rhesus TNFα

.

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

.

23. The isolated human antibody, or an antigen-binding portion thereof, of claim 21, which also neutralizes the activity of pig TNFα

.

26. An isolated human antibody, or antigen-binding portion thereof, that binds to human TNFα

and comprises 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, and

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

29. A pharmaceutical composition comprising the isolated human antibody of any one of claims 26, 27 or 28, and a pharmaceutically acceptable carrier.

30. The pharmaceutical composition of claim 29, which further comprises at least one additional therapeutic agent.

27. An isolated human antibody, or an antigen-binding portion thereof, that binds human TNFα

and comprises 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 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.

28. An isolated human antibody that binds human TNFα

and is the antibody D2E7 or an antigen binding portion thereof.