HUMAN MONOCLONAL ANTIBODIES AGAINST INTERLEUKIN 8 (IL-8)

• US 20120164143A1
• Filed: 12/27/2011
• Published: 06/28/2012
• Est. Priority Date: 12/16/2002
• Status: Active Grant

• Current Assignee: Cormorant Pharmaceuticals AB (Bristol-Myers Squibb Company)
• Original Assignee: Genmab A/S
• Inventors: TEELING, Jessica, PARREN, Paul, BAADSGAARD, Ole, HUDSON, Debra, PETERSEN, Jørgen
• Granted Patent: US 8,603,469 B2
• Time in Patent Office: Days
• US Class Current: 424/134.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• 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 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/10 of the bladder
• 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 17/06 Antipsoriatics
• CPC Class Codes: A61P 17/10 Anti-acne agents
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• 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 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/12 : Antivirals
• CPC Class Codes: A61P 31/16 : for influenza or rhinoviruses
• CPC Class Codes: A61P 31/18 : for HIV
• CPC Class Codes: A61P 31/22 : for herpes viruses
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 35/04 : specific for metastasis
• 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 7/00 : Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/06 : Antianaemics
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• 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 16/244 : Interleukins [IL]
• CPC Class Codes: C07K 16/4241 : against anti-human or anti-animal Ig
• 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 2317/565 : Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/622 : Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: G01N 33/6869 : Interleukin

Abstract

Isolated human monoclonal antibodies which bind to IL-8 (e.g., human IL-8) are disclosed. The human antibodies can be produced in a hybridoma, transfectoma or in a non-human transgenic animal, e.g., a transgenic mouse, capable of producing multiple isotypes of human monoclonal antibodies by undergoing V-D-J recombination and isotype switching. Also disclosed are pharmaceutical compositions comprising the human antibodies, non-human transgenic animals, hybridomas, and transfectomas which produce the human antibodies, and therapeutic and diagnostic methods for using the human antibodies.

Claims

1. A method of treating cancer mediated by human IL-8 comprising administering to a subject an effective amount of a monoclonal antibody which specifically binds to human IL-8 comprising:

(a) a heavy chain variable region comprising an amino acid sequence which is set forth in SEQ ID NO;

12 or which is at least 95% homologous to SEQ ID NO;

12 or which is encoded by a nucleic acid set forth in SEQ ID NO;

10 or encoded by a nucleotide sequence which is at least 95% homologous to SEQ ID NO;

10;

or

(b) a light chain variable region comprising an amino acid sequence which is set forth in SEQ ID NO;

8 or which is at least 95% homologous to SEQ ID NO;

8 or which is encoded by a nucleic acid set forth in SEQ ID NO;

6 or which is encoded by a nucleotide sequence which is at least 95% homologous to SEQ ID NO;

6.

2. The method of claim 1, wherein the antibody comprises human heavy and light chain variable regions comprising amino acid sequences set forth in SEQ ID NOs:

12 and 8, respectively, or amino acid sequences which are at least 95% homologous to SEQ ID NOs;

12 and 8, respectively;

or encoded by nucleic acids comprising nucleotide sequences in their variable regions as set forth in SEQ ID NOs;

10 and 6, respectively;

or encoded by nucleic acids comprising nucleotide sequences which are at least 95% homologous to SEQ ID NOs;

10 and 6, respectively.

17. The method of claim 1, wherein the antibody is selected from the group consisting of an IgG1, an IgG2, an IgG3, an IgG4, an IgM, an IgA1, an IgA2, a secretory IgA, an IgD, and an IgE antibody.

18. The method of claim 17, wherein the antibody is an IgG1 antibody.

19. The method of claim 18, wherein the antibody is an IgG1,κ

or IgG1,λ

isotype.

23. The method of claim 1, wherein the antibody:

(i) inhibits IL-8 binding to its receptors (CXCR1 and CXCR2);

(ii) inhibits IL-8 induced proinflammatory effects;

(iii) inhibits IL-8 induced chemotactic activity for neutrophils;

(iv) inhibits IL-8 induced calcium flux;

(v) inhibits IL-8 induced changes in expression levels of adhesion molecules on neutrophils;

(vi) inhibits IL-8 induced increased expression of CD11b (Mac-1) and inhibits IL-8 induced decreased expression of L-selectin on neutrophils;

(vii) does not cross-react with related chemokines selected from the group consisting of human GRO-α

, human GRO-β

, human IP-10 and human NAP-2;

or

(viii) significantly inhibits chemotaxis induced by biological fluids which contain multiple chemotactic factors including IL-8.

24. The method of claim 1, wherein the antibody has a dissociation equilibrium constant (KD) of approximately 10⁻

8 M or less, when determined by surface plasmon resonance (SPR) technology in a BIACORE 3000 instrument using recombinant human IL-8 as the analyte and the antibody as the ligand.

25. The method of claim 1, wherein the antibody is an intact antibody selected from the group consisting of an intact IgG1 antibody, an intact IgG2 antibody, an intact IgG3 antibody, an intact IgG4 antibody, an intact IgM antibody, an intact IgA1 antibody, an intact IgA2 antibody, an intact secretory IgA antibody, an intact IgD antibody, and an intact IgE antibody, wherein the antibody is glycosylated in a eukaryotic cell.

26. The method of claim 1, wherein the antibody is an antibody fragment or a single chain antibody.

27. The method of claim 1, wherein the antibody is a binding-domain immunoglobulin fusion protein comprising

(i) a variable heavy chain amino acid sequence as set forth in SEQ ID NO:

12 or a variable heavy chain sequence which is at least 95% homologous to SEQ ID NO;

12, a variable light chain amino acid sequence as set forth in SEQ ID NO;

8 or a variable light chain sequence which is at least 95% homologous to SEQ ID NO;

8, or a variable heavy chain amino acid sequence as set forth in SEQ ID NO;

12 or a variable heavy chain sequence which is at least 95% homologous to SEQ ID NO;

12, fused to a variable light chain amino acid sequence as set forth in SEQ ID NO;

8 or a variable light chain sequence which is at least 95% homologous to SEQ ID NO;

8 via a linker peptide, that is fused to an immunoglobulin hinge region polypeptide,

(ii) an immunoglobulin heavy chain CH2 constant region fused to the hinge region, and

(iii) an immunoglobulin heavy chain CH3 constant region fused to the CH2 constant region.

28. The method of claim 1, wherein the cancer is selected from the group consisting of melanoma, thyroid carcinoma, transitional cell carcinoma, trichilemmona, squamous cell carcinoma and breast cancer.

29. The method of claim 1, comprising administering at least one additional therapeutic agent to the subject.

30. The method of claim 29 wherein the agent is selected from the group consisting of coal tar, A vitamin, anthralin, calcipotrien, tarazotene, corticosteroids, methotrexate, retinoids, and hydroxyurea.

31. The method of claim 29, further comprising the step of exposing the subject to sunlight or phototherapy.

32. The method of claim 29, wherein the agent is selected from the group consisting of agents that block or interfere with the function of CC or CXC chemokine receptors, and agents that block the function of chemokine ligands.

33. The method of claim 32, wherein the agent is selected form the group consisting of, an antagonist of CXCR1, an antagonist of CXCR2, an antagonist of CCR1, an antagonist of CCR2, an antagonist of CCR5, an antibody to MIP-1α

, an antibody to MIP-1β

, an antibody to RANTES, an antibody to MCP-1, an antibody to MCP-2, an antibody to MCP-3, and an antibody to MCP-4.

34. The method of claim 1, wherein the antibody is a human, humanized, or chimeric antibody.

3. A method of treating cancer mediated by human IL-8 comprising administering to a subject an effective amount of a monoclonal antibody which specifically binds to human IL-8 antibody comprising at least one CDR sequence selected from the group consisting of:

VL CDR1 of SEQ ID NO;

16, VL CDR2 of SEQ ID NO;

17, VL CDR3 of SEQ ID NO;

18, VH CDR1 of SEQ ID NO;

22, VH CDR2 of SEQ ID NO;

23, and VH of SEQ ID NO;

24.

4. The method of claim 3, wherein the antibody comprises the VH CDR3 of SEQ ID NO:

24.

5. The method of claim 3, wherein the antibody comprises at least four CDR sequences selected from the group consisting of:

VL CDR1 of SEQ ID NO;

16, VL CDR2 of SEQ ID NO;

17, VL CDR3 of SEQ ID NO;

18, VH CDR1 of SEQ ID NO;

22, VH CDR2 of SEQ ID NO;

23, and VH CDR3 of SEQ ID NO;

24.

6. The method of claim 5, wherein the antibody comprises VL CDR1 of SEQ ID NO:

16, VL CDR2 of SEQ ID NO;

17, VL CDR3 of SEQ ID NO;

18, VH CDR1 of SEQ ID NO;

22, VH CDR2 of SEQ ID NO;

23, and VH CDR3 of SEQ ID NO;

24.

35. The method of claim 3, wherein the antibody has a dissociation equilibrium constant (KD) of approximately 10⁻

8 M or less, when determined by surface plasmon resonance (SPR) technology in a BIACORE 3000 instrument using recombinant human IL-8 as the analyte and the antibody as the ligand.

36. The method of claim 3, wherein the cancer is selected from the group consisting of melanoma, thyroid carcinoma, transitional cell carcinoma, trichilemmona, squamous cell carcinoma and breast cancer.

7-8. -8. (canceled)

9. A method of treating cancer mediated by human IL-8 comprising administering to a subject an effective amount of a monoclonal antibody which specifically binds to human IL-8 antibody comprising:

(a) a VL CDR3 domain having the amino acid sequence;

10. The method of claim 9, wherein X1 is different from Gly, X2 is different from Ser, and X3 is different from Pro.

11. The method of claim 10, wherein X1 is Ala, and X2 and X3 independently are Gly, Ala, Val, Leu, or Ile.

12. A method of treating cancer mediated by human IL-8 comprising administering to a subject an effective amount of a monoclonal antibody which specifically binds to a human IL-8 antibody comprising CDR1, CDR2, and CDR3 heavy chain regions derived from the VH 3-33 germline amino acid sequence shown in FIG. 5 (SEQ ID NO:

11) or the CDR1, CDR2, and CDR3 light chain regions derived from the Vκ

A-27 germline amino acid sequence shown in FIG. 3 (SEQ ID NO;

7), and wherein at least one of the CDR domains is selected from the group consisting of;

(a) a light chain CDR1 region comprising an amino acid sequence which is at least 92% identical to the amino acid sequence shown in FIG. 3 (SEQ ID NO;

16);

(b) a light chain CDR2 region comprising an amino acid sequence which is at least 86% identical to the amino acid sequence shown in FIG. 3 (SEQ ID NO;

17);

(c) a light chain CDR3 region comprising an amino acid sequence which is at least 43% identical to the amino acid sequence shown in FIG. 3 (SEQ ID NO;

18);

(d) a heavy chain CDR1 region comprising an amino acid sequence which is at least 61% identical to the amino acid sequence shown in FIG. 5 (SEQ ID NO;

22);

(e) a heavy chain CDR2 region comprising an amino acid sequence which is at least 77% identical to the amino acid sequence shown in FIG. 5 (SEQ ID NO;

23); and

(f) a heavy chain CDR3 region comprising an amino acid sequence which is at least 76% identical to the amino acid sequence shown in FIG. 5 (SEQ ID NO;

24); and

(g) any combination of (a), (b), (c), (d), (e), or (f).

13. The method of claim 12, wherein the antibody comprises:

(a) a CDR1 human heavy chain region comprises a histidine and a tyrosine residue at positions 1 and 5, respectively, shown in FIG. 5 (SEQ ID NO;

22);

(b) a CDR2 human heavy chain region comprises an isoleucine, tyrosine, asparagine, and alanine residue at positions 2, 8, 11, and 12, respectively, shown in FIG. 5 (SEQ ID NO;

23);

(c) a CDR3 human heavy chain region comprises an arginine and leucine residue at positions 2 and 5, respectively, shown in FIG. 5 (SEQ ID NO;

24);

(d) a CDR1 human light chain region comprises an isoleucine residue at position 6, shown in FIG. 3 (SEQ ID NO;

16);

(e) a CDR2 human light chain region comprises a proline residue at position 2, shown in FIG. 3 (SEQ ID NO;

17); and

(f) a CDR3 human light chain region comprises a tyrosine, alanine, glycine, and leucine residue at positions 3, 4, 5, and 7, respectively, shown in FIG. 3 (SEQ ID NO;

18).

14. The method of claim 12, wherein the antibody comprises:

(a) a light chain variable region having an amino acid sequence which is at least 94% identical to the Vκ

A-27 germline amino acid sequence shown in FIG. 3 (SEQ ID NO;

7) or

(b) a heavy chain variable region having an amino acid sequence which is at least 92% identical to the VH 3-33 germline amino acid sequence shown in FIG. 5 (SEQ ID NO;

11).

15. The method of claim 14, wherein the antibody comprises a light chain variable region derived from the Vκ

A-27 germline amino acid sequence shown in FIG. 3 (SEQ ID NO;

7) and having an amino acid sequence which comprises at least one residue selected from the group consisting of an isoleucine at position 29, a proline residue at position 52, an alanine residue at position 93, a glycine residue at position 94, a leucine residue at position 96, a proline residue at position 100, an aspartic acid at position 105, and any combination thereof.

16. The method of claim 14 wherein the antibody comprises a heavy chain variable region derived from the VH 3-33 germline amino acid sequence shown in FIG. 5 (SEQ ID NO:

11) and having an amino acid sequence which comprises at least one residue selected from the group consisting of a glutamine at position 3, a histidine residue at position 31, a tyrosine residue at position 35, an isoleucine residue at position 51, a tyrosine residue at position 57, an asparagine residue at position 60, an alanine residue at position 61, an isoleucine residue at position 70, an asparagine residue at position 74, a glutamine residue at position 82, an arginine residue at position 100, a leucine residue at position 103, and any combination thereof.

20-22. -22. (canceled)