Compositions and methods of use for antibodies of c-Met

• US 8,101,727 B2
• Filed: 03/29/2007
• Issued: 01/24/2012
• Est. Priority Date: 03/30/2006
• Status: Active Grant

• Current Assignee: Novartis Ag
• Original Assignee: Novartis Ag
• Inventors: Stover, David Raymond, Prassler, Josef, Berger, Catrin, Brocks, Bodo
• Primary Examiner(s): Wen, Sharon
• Application Number:
  US12/295,535
• Publication Number:
  US 20090175860A1
• Time in Patent Office: 1,762 Days
• Field of Search: None
• US Class Current: 530/388.22
• 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 35/02 specific for leukemia
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: C07K 16/2863 against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• 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 2317/567 Framework region [FR]
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• 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: Y10S 435/81 Packaged device or kit

Abstract

Antibodies and fragments that bind to the protein target c-Met, particularly to epitopes located in the c-Met extracellular domain, are provided, as are methods of use of the antibodies and kits, for treating an unwanted cell, in particular, a cell associated with a c-Met-related condition such as a cancer, a metastasis, or an inflammatory condition.

Claims

1. An isolated human or humanized antibody or functional fragment thereof comprising an antigen-binding region that is specific for target protein c-Met, wherein the antibody or functional fragment thereof binds to c-Met, and wherein the sequence of the antibody or functional fragment thereof comprises:

the sequences of CDR1, CDR2 and CDR3 of heavy chain 4687, wherein the sequences of CDR1, CDR2, and CDR3 of heavy chain 4687 are residues 26-35, 50-65, and 98-102, respectively, of SEQ ID NO;

58; and

the sequences of CDR1, CDR2, and CDR3 of light chain 5097, wherein the sequences of CDR1, CDR2, and CDR3 of light chain 5097 are residues 24-39, 55-61, and 94-100 of SEQ ID NO;

37.

2. The antibody or functional fragment thereof according to claim 1, wherein the antibody or functional fragment thereof binds the target protein c-Met with a K_D of 2.0×

10<sup>−

5</sup> M or less, 2.0×

10<sup>−

6</sup> M or less, 2.0×

10<sup>−

7</sup> M or less, 2.0×

10<sup>−

8</sup> M or less, and 2.0×

10<sup>−

9</sup> M or less.

3. The antibody or functional fragment thereof according to claim 1, wherein the antibody or functional fragment thereof has an off rate (K_off) for target protein c-Met of 1.0×

10<sup>−

2</sup> per sec or smaller, 1.0×

10<sup>−

3</sup> per sec or smaller, 1×

10<sup>−

4</sup> per sec or smaller or 1.0×

10<sup>−

5</sup> per sec or smaller.

4. The antibody or functional fragment thereof according to claim 1, wherein the antibody or functional fragment thereof binds the target protein c-Met with a K_D of 2.0×

10<sup>−

5</sup> M or less, 2.0×

10<sup>−

6</sup> M or less, 2.0×

10<sup>−

7</sup> M or less, 2.0×

10<sup>−

8</sup> M or less, and 2.0×

10<sup>−

9</sup> M or less, and inhibits HGF binding to c-Met.

5. The antibody or functional fragment thereof according to claim 1, wherein the antibody or functional fragment thereof binds the target protein c-Met and modulates c-Met phosphorylation.

6. The antibody or functional fragment thereof according to claim 5, wherein activating c-Met phosphorylation stimulates at least one of an activity selected from the group of organ regeneration, wound healing, and tissue regeneration.

7. The antibody or functional fragment thereof according to claim 6, wherein the organ is selected from the group of kidney, liver, pancreas, heart, lung, intestine, skin, thymus, and thyroid.

8. The antibody according to claim 1, wherein binding of the antibody to c-Met is determined by at least one assay selected from an amount of antagonism or agonism of:

ligand induction of a c-Met signal transduction pathway enzyme activity;

ligand induction of a c-Met signal transduction pathway gene expression;

electrochemiluminescence-based binding of a ligand to c-Met;

enzyme-linked immunosorbent assay of binding of a ligand to c-Met; and

proliferation, survival, migration or metastasis of a cell.

9. An isolated functional fragment according to claim 1.

14. The isolated antibody according to claim 1, which is an IgG.

15. The isolated antibody according to claim 14, which is an IgG1, an IgG2, an IgG3 or an IgG4.

16. The isolated antibody or functional fragment according to claim 1 that is a Fab or scFv antibody fragment.

17. The isolated antibody or functional fragment thereof according to claim 1, comprising an antigen-binding region that is specific for an epitope of c-Met, wherein the epitope is a conformational epitope.

18. The isolated antibody or functional fragment thereof according to claim 17 wherein the conformational epitope comprises residues of an amino acid sequence of an extracellular domain of c-Met.

19. A pharmaceutical composition comprising at least one antibody or functional fragment according to claim 1 and a pharmaceutically acceptable carrier or excipient therefore.

20. A human or humanized antibody or antibody fragment thereof as described in claim 1, wherein the antibody is a synthetic antibody.

21. The pharmaceutical composition according to claim 19, further comprising an additional therapeutic agent.

22. The pharmaceutical composition according to claim 21, wherein the additional therapeutic agent is selected from the group consisting of an anti-cancer agent;

an antibiotic;

an anti-inflammatory agent;

a growth factor; and

a cytokine.

23. An immunoconjugate comprising a first component which is an antibody or fragment thereof according to claim 1.

24. The immunoconjugate according to claim 23, comprising a second component having a second amino acid sequence.

25. The immunoconjugate according to claim 24, further comprising a cytotoxin.

26. The immunoconjugate according to claim 24, wherein the second component is a binding protein or antibody having a binding specificity for a target that is different from c-Met.

27. A bispecific antibody comprising an immunoconjugate comprising a first component comprising a functional fragment according to claim 1.

28. The bispecific antibody according to claim 27, wherein the target of the binding specificity different from c-Met is a tumor antigen or tumor-associated protein on a surface of a cancer cell.

29. A kit comprising an antibody or fragment thereof according to claim 1.

30. The kit according to claim 29, further comprising a pharmaceutically acceptable carrier or excipient therefore.

31. The kit according to claim 29 wherein the antibody is present in a unit dose and further comprising instructions for use in administering to a subject.

10. An isolated amino acid sequence that binds to and is specific for target protein c-Met comprising CDR1, CDR2 and CDR3 sequences of the heavy and light chains of an antigen-binding region that is specific for target protein c-Met, wherein the CDR1, CDR2 and CDR3 of the heavy and light chains are encoded by nucleotides 78-105, 150-195, and 294-306, respectively, of SEQ ID NO:

24 and nucleotides 72-117, 165-183, and 282-300, respectively, of SEQ ID NO;

7.

11. The isolated amino acid sequence of claim 10 comprising an antigen-binding region, said region comprising a light chain encoded by a nucleotide sequence of SEQ ID NO:

7.

12. The isolated amino acid sequence of claim 10 comprising an antigen-binding region, said region comprising a heavy chain encoded by a nucleotide sequence of SEQ ID NO:

24.

13. An isolated amino acid sequence comprising CDR1, CDR2 and CDR3 sequences of a light chain and a heavy chain of an antigen-binding region that is specific for target protein c-Met, wherein the CDR sequences from the heavy chain are:

CDR1=GGTFSSYAIS (residues 26-35 of SEQ ID NO;

     58)

CDR2=GIDPFGTANYAQKFQG (residues 50-65 of SEQ ID NO;

     58)

CDR3=VYQDV (residues 98-102 of SEQ ID NO;

     58), and the CDR sequences from the light chain are;

CDR1=RSSQSILYGINNNFLG (residues 24-39 of SEQ ID NO;

     37)

CDR2=WASTRES (residues 55-61 of SEQ ID NO;

     37)

CDR3=QQYAFGW (residues 94-100 of SEQ ID NO;

     37).