Anti-HER2 antibody variants

• US 7,435,797 B2
• Filed: 04/09/2003
• Issued: 10/14/2008
• Est. Priority Date: 04/10/2002
• Status: Active Grant

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Lowman, Henry B., Carter, Paul J., Gerstner, Resi B.
• Primary Examiner(s): Blanchard; David J.
• Application Number:
  US10/410,894
• Publication Number:
  US 20030228663A1
• Time in Patent Office: 2,015 Days
• Field of Search: 530/387.3, 530/350, 530/388.2, 530/326, 530/300, 530/387.1, 530/387.5, 530/387.7, 424/133.1, 424/130.1, 424/143.1, 424/277.1
• US Class Current: 530/387.3
• CPC Class Codes: A61J 1/00 Containers specially adapted for medical or pharmaceutical purposes capsules or the like for oral use A61J3/07; specially adapted for surgical or diagnostic appliances or instruments A61B50/30; containers for radioactive substances G21F5/00
• CPC Class Codes: A61J 1/18 Arrangements for indicating condition of container contents, e.g. sterile condition
• CPC Class Codes: A61J 2205/30 Printed labels
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: C07K 16/3015 Breast
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• 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 2317/54 F(ab')2
• 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 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Abstract

The present invention concerns novel antibody variants, particularly anti-HER2 antibody variants having substitutions at positions within the variable domains of the heavy and light chains

Claims

1. An isolated antibody that is capable of binding to the extracellular domain of HER2, which comprises an antibody light chain variable domain of SEQ ID NO:

1, and an antibody heavy chain variable domain of SEQ ID NO;

2, wherein the light chain variable domain of SEQ ID NO;

1 comprises an amino acid substitution selected from the group consisting of N30(V_L)S;

T31(V_L)S;

Y49(V_L)D, and Y49(V_L)W numbered according to the Kabat numbering system, and the heavy chain variable domain of SEQ ID NO;

2 comprises an amino acid substitution selected from the group consisting of D98(V_H)W, F100(V_H)P;

Y102(V_H)K; and

Y102(V_H)L, numbered according to the Kabat numbering system.

2. The antibody of claim 1 wherein the amino acid substitution in the light chain variable domain is Y49(V_L)D.

3. The antibody of claim 1, wherein the amino acid substitution in the light chain variable domain is N30(V_L)S.

5. The antibody of claim 1, which is a humanized antibody.

6. The antibody of claim 1, which is a antibody fragment selected from the group consisting of Fv, Fab, Fab′

and F(ab′

)₂ fragments.

7. The antibody of claim 1 wherein the amino acid substitution in the heavy chain variable domain is D98(V_H)W.

8. The antibody of claim 1 wherein the amino acid substitution in the heavy chain variable domain is F100(V_H)P or Y102(V_H)K.

9. The antibody of claim 1 wherein the amino acid substitution in the heavy chain variable domain is F100(V_H)P or Y102(V_H)L.

4. An isolated antibody that is capable of binding to the extracellular domain of HER2, which comprises an antibody light chain variable domain of SEQ ID NO:

1, wherein N30(V_L) is substituted with S, H91(V_L) is substituted with F, and Y92(V_L) is substituted with W, numbered according to the Kabat numbering system, and an antibody heavy chain variable domain of SEQ ID NO;

2, comprising an amino acid substitution selected from the group consisting of substitutions D98(V_H)R, F100(V_H)P and Y102(V_H)K, numbered according to the Kabat numbering.

10. An isolated antibody that is capable of binding to the extracellular domain of HER2, which comprises an antibody light chain variable domain of SEQ ID NO:

1, and an antibody heavy chain variable domain of SEQ ID NO;

2, wherein said light chain variable domain comprises an amino acid substitution selected from the group consisting of Y49(V_L)D, F53(V_L)W, and Y55(V_L)W, numbered according to the Kabat numbering system.

12. The antibody of claim 10 or 11, which is a humanized antibody.

13. The antibody of claim 10 or 11, which is an antibody fragment selected from the group consisting of Fv, Fab, Fab′

and F(ab′

)₂ fragments.

11. An isolated antibody that is capable of binding to the extracellular domain of HER2, which comprises an antibody light chain variable domain of SEQ ID NO:

1 and an antibody heavy chain variable domain of SEQ ID NO;

2, wherein in said light chain variable region N30(V_L) is substituted with S, H91(V_L) is substituted with F, and Y92(V_L) is substituted with W.

14. An isolated antibody that is capable of binding to the extracellular domain of HER2, which comprises an antibody light chain variable domain of SEQ ID NO:

1 and an antibody heavy chain variabic domain of SEQ ID NO;

2, wherein said heavy chain variable domain comprises amino acid substitutions F100(V_H)P and Y102(V_H)K, numbered according to the Kabat numbering.

15. The antibody of claim 14, which is a humanized antibody.

16. The antibody of claim 14, which is an antibody fragment selected from the group consisting of Fv, Fab, Fab′

and F(ab′

)₂ fragments.

17. An antibody that is capable of binding to the extracellular domain of HER2, which comprises an antibody light chain variable domain of SEQ ID NO:

1 and an antibody heavy chain variable domain of SEQ ID NO;

2, wherein in the light chain variable domain N30(V_L) is substituted with S, H91(V_L) is substituted with F, Y92(V_L) is substituted with W, T94(V_L) is substituted with S, and in the heavy chain variable domain D98(V_H) is substituted with W, Y100a(V_H) is substituted with F, and Y102(V_H) is substituted with V, numbered according to the Kabat numbering system.

18. An isolated antibody that is capable of binding to the extracellular domain of HER2, which comprises comprising an antibody light chain variable domain of SEQ ID NO:

1 and an antibody heavy chain variable domain of SEQ ID NO;

2 with the following substitutions;

Y49(V_L)D, F53(V_L)W, Y55(V_L)W, F100(V_H)P, and Y102(V_H)L.

19. The antibody of claim 18 wherein the heavy chain variable domain of SEQ ID NO:

2 further comprises the substitution D98(V_H)W.

22. The antibody of claim 18, 19, 20, or 21, wherein the binding affinity of the antibody for the HER2 extracellular domain is at least three-fold better than the binding affinity of humanized monoclonal antibody 4D5-8 for the HER2 extracellular domain.

23. The antibody of claim 18, 19, 20, or 21, which is a humanized antibody.

24. The antibody of claim 18, 19, 20, or 21, which is an antibody fragment selected from the group consisting of Fv, Fab, Fab′

and F(ab′

)₂ fragments.

28. An article of manufacture comprising a container, a composition contained therein, and a package insert or label indicating that the composition can be used to treat cancer characterized by the overexpression of HER2, wherein the composition comprises the antibody of claim 26 or 22.

29. The article of manufacture of claim 28, wherein the cancer is breast cancer.

20. An isolated antibody that is capable of binding to the extracellular domain of HER2, which comprises an antibody light chain variable domain of SEQ ID NO:

1 and an antibody heavy chain variable domain of SEQ ID NO;

2 with the following substitutions;

Y49(V_L)D, F53(V_L)W, Y55(V_L)W, F100(V_H)P, and Y102(V_H)K.

21. The antibody of claim 20 wherein the the heavy chain variable domain of SEQ ID NO:

2 further comprises the substitution D98(V_H)W.

25. An isolated antibody that is capable of binding to the extracellular domain of HER2, which comprises an antibody ligbt chain variable domain of SEQ ID NO:

1 and an antibody heavy chain variable domain of SEQ ID NO;

2, wherein N30(V_L) is substituted with S, H91(V_L) is substituted with F, and Y92(V_L) is substituted with W, numbered according to the Kabat numbering system.

26. A humanized anti-HER2 antibody 4D5-8, comprising one or more amino acid substitutions selected from the group consisting of N30(V_L)S, Y49(V_L)W, F53(V_L)W, H91(V_L)F, Y92(V_L)W, and D98(V_H)W, numbered according to the Kabat numbering system.

27. A humanized anti-HER2 antibody 4D5-8, which comprises a combination of amino acid substitutions selected from the group consisting of (i) Y95(V_L)W, F53(V_L)V, H91(V_L)F and D98(V_H)R, F100(V_H)L;

(ii) Y49(V_L)D, F53(V_L)W, Y55(V_L)W, F100(V_H)P, Y102(V_H)K;

(iii) Y49(V_L)D, F53(V_L)W, Y55(V_L)W, F100(V_H)P, Y102(V_H)L;

(iv) Y49(V_L)V, F53(V_L)V, Y55(V_L)W, F100(V_H)W, Y102(V_H)L;

(v) Y49(V_L)W, F53(V_L)O, Y55(V_L)F, Y102(V_H)W;

(vi) Y49(V_L)D, F53(V_L)W, Y55(V_L)W, F100(V_H)P, Y102(V_H)K;

(vii) D28(V_L)G, N30(V_L)S, T31(V_L)S, R66(V_L)N, Y92(V_L)W, F100(V_H)P, Y102(V_H)K; and

(viii) D28(V_L)G, N30(V_L)S, T31(V_L)S, R66(V_L)M, Y92(V_L)F, F100(V_H)P, Y102(V_H)L.

30. An isolated antibody that is capable of binding to the extracellular domain of HER2, which comprises an antibody light chain variable domain of SEQ ID NO:

1 and an antibody heavy chain variable domain of SEQ ID NO;

2 wherein the heavy chain variable domain comprises an amino acid substitution 98(V_H)W.