Polypeptide variants with altered effector function

• US 20060194957A1
• Filed: 05/08/2006
• Published: 08/31/2006
• Est. Priority Date: 01/15/1999
• Status: Active Grant

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Presta, Leonard
• Granted Patent: US 7,335,742 B2
• Time in Patent Office: Days
• US Class Current: 530/388.22
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/2839 against the integrin superfamily
• CPC Class Codes: C07K 16/2887 against CD20
• CPC Class Codes: C07K 16/4291 against IgE
• CPC Class Codes: C07K 2317/14 Specific host cells or culture conditions, e.g. components, pH or temperature
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/524 CH2 domain
• CPC Class Codes: C07K 2317/71 Decreased effector function due to an Fc-modification
• CPC Class Codes: C07K 2317/732 Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2317/734 Complement-dependent cytotoxicity [CDC]
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C12N 2799/022 where the vector is derived from an adenovirus

Abstract

The present invention concerns polypeptides comprising a variant Fc region. More particularly, the present invention concerns Fc region-containing polypeptides that have altered effector function as a consequence of one or more amino acid modifications in the Fc region thereof.

Claims

1. A variant of a parent polypeptide comprising an Fc region, which variant mediates antibody-dependent cell-mediated cytotoxicity (ADCC) in the presence of human effector cells more effectively, or binds an Fc gamma receptor (Fcγ

R) with better affinity, than the parent polypeptide and comprises at least one amino acid modification in the Fc region.

2. The variant of claim 1 which comprises an antibody.

3. The variant of claim 1 wherein the parent polypeptide Fc region comprises a human IgG Fc region.

4. The variant of claim 3 wherein the human IgG Fc region comprises a human IgG1, IgG2, IgG3 or IgG4 Fc region.

5. The variant of claim 1 which mediates ADCC about 1.5 fold to about 100 fold more effectively than the parent polypeptide.

6. The variant of claim 1 which binds an Fcγ

RIII with better affinity than the parent polypeptide.

7. The variant of claim 6 which further binds an Fcγ

RII with worse affinity than the parent polypeptide.

8. The variant of claim 1 which comprises at least one amino acid substitution in the Fc region.

9. The variant of claim 1 which comprises at least one amino acid modification in a CH2 domain of the Fc region.

10. The variant of claim 1 which comprises at least one amino acid modification in the Fc region, other than in a lower hinge region thereof.

11. The variant of claim 1 which comprises an amino acid substitution at any one or more of amino acid positions 256, 290, 298, 312, 326, 330, 333, 334, 360, 378 or 430 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.

12. The variant of claim 11 which comprises two or more amino acid substitutions at the amino acid positions listed therein.

13. The variant of claim 11 which comprises three or more amino acid substitutions at the amino acid positions listed therein.

36. A composition comprising the polypeptide variant of claim 1 and a pharmaceutically acceptable carrier.

37. The composition of claim 36 which is sterile.

38. Isolated nucleic acid encoding the polypeptide variant of claim 1.

39. A vector comprising the nucleic acid of claim 38.

40. A host cell containing the vector of claim 39.

41. A method for producing a polypeptide variant comprising culturing the host cell of claim 40 so that the nucleic acid is expressed.

42. The process of claim 41 further comprising recovering the polypeptide variant from the host cell culture.

43. A method for treating a disorder in a mammal comprising administering to the mammal a therapeutically effective amount of the polypeptide variant of claim 1.

14. A polypeptide comprising a variant Fc region with altered Fc gamma receptor (Fcγ

R) binding affinity, which polypeptide comprises an amino acid modification at any one or more of amino acid positions 238, 239, 248, 249, 252, 254, 255, 256, 258, 265, 267, 268, 269, 270, 272, 276, 278, 280, 283, 285, 286, 289, 290, 292, 293, 294, 295, 296, 298, 301, 303, 305, 307, 309, 312, 315, 320, 322, 324, 326, 327, 329, 330, 331, 333, 334, 335, 337, 338, 340, 360, 373, 376, 378, 382, 388, 389, 398, 414, 416, 419, 430, 434, 435, 437, 438 or 439 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.

15. The polypeptide of claim 14 wherein the variant Fc region comprises a variant human IgG Fc region.

16. The polypeptide of claim 14 which displays reduced binding to an Fcγ

R and comprises an amino acid modification at any one or more of amino acid positions 238, 239, 248, 249, 252, 254, 265, 268, 269, 270, 272, 278, 289, 292, 293, 294, 295, 296, 298, 301, 303, 322, 324, 327, 329, 333, 335, 338, 340, 373, 376, 382, 388, 389, 414, 416, 419, 434, 435, 437, 438 or 439 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.

17. The polypeptide of claim 14 which displays reduced binding to an Fcγ

RI.

18. The polypeptide of claim 17 which displays reduced binding to the Fcγ

RI and comprises an amino acid modification at any one or more of amino acid 238, 265, 269, 270, 327 or 329 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.

19. The polypeptide of claim 14 which displays reduced binding to an Fcγ

RII.

20. The polypeptide of claim 19 which displays reduced binding to the Fcγ

RII and comprises an amino acid modification at any one or more of amino acid positions 238, 265, 269, 270, 292, 294, 295, 298, 303, 324, 327, 329, 333, 335, 338, 373, 376, 414, 416, 419, 435, 438 or 439 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.

21. The polypeptide of claim 14 which displays reduced binding to an Fcγ

RIII.

22. The polypeptide of claim 21 which displays reduced binding to the Fcγ

RIII and comprises an amino acid modification at any one or more of amino acid 238, 239, 248, 249, 252, 254, 265, 268, 269, 270, 272, 278, 289, 293, 294, 295, 296, 301, 303, 322, 327, 329, 338, 340, 373, 376, 382, 388, 389, 416, 434, 435 or 437 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.

23. The polypeptide of claim 14 which displays increased binding to an Fcγ

R and comprises an amino acid modification at any one or more of amino acid positions 255, 256, 258, 267, 268, 272, 276, 280, 283, 285, 286, 290, 298, 301, 305, 307, 309, 312, 315, 320, 322, 326, 330, 331, 333, 334, 337, 340, 360, 378, 398 or 430 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.

24. The polypeptide of claim 23 which displays increased binding to an Fcγ

RIII.

25. The polypeptide of claim 24 which further displays decreased binding to an Fcγ

RII.

26. The polypeptide of claim 25 which displays increased binding to the Fcγ

RIII and further displays decreased binding to the Fcγ

RII, wherein the polypeptide comprises an amino acid modification at positions 298 and/or 333 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.

27. The polypeptide of claim 23 which displays increased binding to an Fcγ

RII.

28. The polypeptide of claim 27 which displays increased binding to the Fcγ

RII and comprises an amino acid modification at any one or more of amino acid 255, 256, 258, 267, 268, 272, 276, 280, 283, 285, 286, 290, 301, 305, 307, 309, 312, 315, 320, 322, 326, 330, 331, 337, 340, 378, 398 or 430 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.

29. The polypeptide of claim 27 which further displays decreased binding to an Fcγ

RIII.

30. The polypeptide of claim 29 which displays increased binding to the Fcγ

RII and further displays decreased binding to the Fcγ

RIII, wherein the polypeptide comprises an amino acid modification at any one or more of amino acid positions 268, 272, 298, 301, 322 or 340 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.

31. A polypeptide comprising a variant Fc region with altered neonatal Fc receptor (FcRn) binding affinity, which polypeptide comprises an amino acid modification at any one or more of amino acid positions 238, 252, 253, 254, 255, 256, 265, 272, 286, 288, 303, 305, 307, 309, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 386, 388, 400, 413, 415, 424, 433, 434, 435, 436, 439 or 447 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.

32. The polypeptide of claim 31 which displays reduced binding to an FcRn.

33. The polypeptide of claim 32 which displays reduced binding to the FcRn and comprises an amino acid modification at any one or more of amino acid positions 252, 253, 254, 255, 288, 309, 386, 388, 400, 415, 433, 435, 436, 439 or 447 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.

34. The polypeptide of claim 31 which displays increased binding to FcRn.

35. The polypeptide of claim 34 which displays increased binding to FcRn and comprises an amino acid modification at any one or more of amino acid positions 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424 or 434 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.

44. A method for making a variant Fc region with altered Fc receptor (FcR) binding affinity, or altered antibody-dependent cell-mediated cytotoxicity (ADCC) activity, comprising:

(a) introducing one or more amino acid modifications into an Fc region of a parent polypeptide in order to generate a variant Fc region;

(b) determining binding of the variant Fc region to an FcR, or determining ADCC activity of the variant Fc region.

45. The method of claim 44 wherein step (b) comprises determining binding of the variant Fc region to an FcR in vitro.

46. The method of claim 44 wherein step (b) comprises identifying a variant Fc region with improved FcR binding affinity, or with improved ADCC activity.

47. The method of claim 44 wherein the FcR is human Fc gamma receptor III (Fcγ

RIII).

48. The method of claim 44 wherein step (b) comprises determining binding of the variant Fc region to at least two different FcRs.

49. The method of claim 48 wherein the Fc receptors include human Fc gamma receptor II (Fcγ

RII) and human Fc gamma receptor III (Fcγ

RIII).