Humanized anti-TGF-beta antibodies

• US 7,527,791 B2
• Filed: 03/31/2005
• Issued: 05/05/2009
• Est. Priority Date: 03/31/2004
• Status: Active Grant

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Ferrara, Napoleone, Presta, Leonard G., Adams, Camellia W., Mao, Weiguang, Tejada, Max L., Filvaroff, Ellen H.
• Primary Examiner(s): Rao; Manjunath
• Assistant Examiner(s): Shafer; Shulamith H
• Application Number:
  US11/096,046
• Publication Number:
  US 20050276802A1
• Time in Patent Office: 1,496 Days
• Field of Search: None
• US Class Current: 424/145.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• 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 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• 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 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• 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: Y10S 435/81 Packaged device or kit

Abstract

Humanized anti-TGF-beta antibodies are provided, as well as methods for their preparation and use, including methods for treating TGF-beta disorders, for example, cancer. Also provided are articles of manufacture designed for various uses that contain the humanized antibodies.

Claims

1. A humanized antibody that binds a TGF-beta1, TGF-beta2 or TGF-beta3 protein, the humanized antibody comprising:

a variable heavy (V_H) domain that comprises the non-human hypervariable region residues of SEQ ID NO;

21, SEQ ID NO;

23 and SEQ ID NO;

43 incorporated into a human V_H domain, said variable domain further comprising a framework region (FR) comprising the framework residues of SEQ ID NO;

6 and comprising one or more FR substitutions in SEQ ID NO;

6 at a position selected from the group consisting of 48, 49, 68, and 72; and

comprising variable light (V_L) domain complementarity-determining-region (CDR) residues RASQSVLYSSNQKNYLA (SEQ ID NO;

18) or RASQGISSYLA (SEQ ID NO;

7);

WASTRES (SEQ ID NO;

19) or YASSLQS (SEQ ID NO;

8); and

HQYLSSDT (SEQ ID NO;

20).

2. The humanized antibody of claim 1 comprising FR substitutions at positions 49, 68, and 72.

3. The humanized antibody of claim 2 wherein at position 49 the alanine is changed to a glycine, at position 68 phenylalanine is changed to an alanine, and at position 72 the arginine is changed to an alanine.

4. The humanized antibody of claim 1 comprising FR substitutions at positions 48, 49, and 72.

5. The humanized antibody of claim 4 wherein at position 48 the valine is changed to an isolcucine, at position 49 the alanine is changed to a glycine, and at position 72 the arginine is changed to an alanine.

6. The humanized antibody of claim 1 comprising FR substitutions at positions 49, 70, and 72.

7. The humanized antibody of claim 6 wherein at position 49 the alanine is changed to a glycine, at position 70 the isoleucine is changed to a leucine, and at position 72 the arginine is changed to an alanine.

8. The humanized antibody of claim 6 wherein an additional FR substitution is at position 74.

9. The humanized antibody of claim 8 wherein at position 74 the asparagine is changed to a lysine.

10. The humanized antibody of claim 1 comprising FR substitutions at positions 49, 72, and 74.

11. The humanized antibody of claim 10 wherein at position 49 the alanine is changed to a glycine, at position 72 the arginine is changed to an alanine, and at position 74 the asparagine is changed to a lysine.

12. The humanized antibody of claim 1 comprising FR substitutions at positions 49, 72, and 79.

13. The humanized antibody of claim 12 wherein at position 49 the alanine is changed to a glycine, at position 72 the arginine is changed to an alanine, and at position 79 the leucine is changed to an alanine.

14. The humanized antibody of claim 1 comprising variable light (V_L) domain complementarity-determining-region (CDR) residues RASQSVLYSSNQKNYLA (SEQ ID NO:

18);

WASTRES (SEQ ID NO;

19); and

HQYLSSDT (SEQ ID NO;

20).

15. The humanized antibody of claim 1 comprising the V_L domain amino acid sequence in SEQ ID NO:

3.

16. The humanized antibody of claim 1 comprising the V_H domain amino acid sequence in SEQ ID NO:

4.

17. The humanized antibody of claim 1 that is an intact IgG1 antibody.

18. The humanized antibody of claim 1 that is an antibody fragment.

19. The humanized antibody of claim 18 that is a Fab fragment.

20. The humanized antibody of claim 1 that is not conjugated with a cytotoxic agent.

21. The humanized antibody of claim 1 that is conjugated with a cytotoxic agent.

22. A composition comprising the humanized antibody of claim 1 and a carrier.

23. An article of manufacture comprising a container containing the humanized antibody of claim 1 and instructions directing a user to treat a TGF-beta disorder in a mammal with the antibody in an effective amount, wherein the TGF-beta disorder is a colo-rectal cancer, a melanoma or a cancer of the breast, prostate or lung.

24. The article of claim 23 additionally comprising a container containing a therapeutic agent other than the humanized antibody, wherein the instructions direct the user to treat the disorder with the antibody in combination with the agent in effective amounts.

25. The article of claim 23 wherein the mammal is a human.

26. The humanized antibody of claim 1, further comprising a FR substitution at positions 70, 74 and 79.