Polypeptide variants with altered effector function

• US 6,737,056 B1
• Filed: 01/14/2000
• Issued: 05/18/2004
• Est. Priority Date: 01/15/1999
• Status: Expired due to Term

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Presta, Leonard
• Primary Examiner(s): SAUNDERS, DAVID A
• Application Number:
  US09/483,588
• Time in Patent Office: 1,586 Days
• Field of Search: 424/133.1, 530/387.3
• US Class Current: 424/133.1
• 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 a human IgG1 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 III (Fcγ

RIII) with better affinity, than the parent polypeptide and comprises two or more amino acid substitutions in the Fc region.

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

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

4. The variant of claim 1 which binds an Fcγ

RIII with better affinity than the parent polypeptide.

5. The variant of claim 4 which further binds an Fcγ

RII with worse affinity than the parent polypeptide.

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

7. The variant of claim 1 which comprises at least one amino acid modification in the Fc region, other than at amino acid positions 233 to 239.

8. The variant of claim 1 which comprises an amino acid substitution at any one or more of amino acid positions 256, 290, 298, 312, 356, 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.

9. The variant of claim 8 which comprises two or more amino acid substitutions at the amino acid positions listed therein.

10. The variant of claim 8 which comprises three or more amino acid substitutions at the amino acid positions listed therein.

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

12. The composition of claim 11 which is sterile.

13. A polypeptide comprising a variant Fc region which is not a native sequence Fc region and has increased binding to an Fc gamma receptor (Fcγ

R), 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, 279, 280, 283, 285, 298, 289, 290, 292, 293, 294, 295, 296, 298, 301, 303, 305, 307, 312, 315, 324, 327, 329, 330, 335, 337, 3338, 340, 360, 373, 376, 379, 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.

14. The polypeptide of claim 13 wherein the variant Fc region comprises a variant human IgG Fc region.

15. The polypeptide of claim 13 which display 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, 280, 283, 285, 286, 290, 298, 301, 305, 307, 312, 315, 330, 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.

16. The polypeptide of claim 15 which displays increased binding to an Fcγ

RIII.

17. The polypeptide of claim 16 which further displays decreased binding to an Fcγ

RII.

18. The polypeptide of claim 17 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 position 298 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 15 which displays increased binding to an Fcγ

RII.

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

RII and comprises an amino acid modification at any one or more of amino acid positions 255, 256, 258, 267, 268, 272, 280, 283, 285, 286, 290, 301, 305, 307, 312, 315, 330, 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.

21. The polypeptide of claim 19 which further displays decreased binding to an Fcγ

RIII.

22. The polypeptide of claim 21 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.

23. A polypeptide comprising a variant Fc region which is not a native sequence Fc region and has increased binding to an Fc gamma receptor (Fcγ

R), which polypeptide comprises an amino acid modification at any one or more of amino acid position 256, 290, 298, 312, 330, 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.

24. The polypeptide of claim 23 which comprises two or more amino acid substitutions at the amino acid positions listed therein.

25. The polypeptide of claim 23 which comprises three or more amino acid substitutions at the amino acid positions listed therein.

26. The polypeptide of claim 23 which comprises an antibody.

27. The polypeptide of claim 23 which comprises an amino acid substitution at position 298.

28. The polypeptide of claim 23 which comprises amino acid substitutions at two or three of positions 298, 333 and 334.

29. The polypeptide of claim 28 which comprises amino acid substitutions at positions 298, 333, and 334.

30. The polypeptide of claim 24 which consists of amino acid substitutions at positions 298, 333, and 334.

31. The polypeptide of claim 23 which comprises a variant human IgG1, IgG2, IgG3 or IgG4 Fc region.

32. The polypeptide of claim 31 which comprises a variant human IgG1 Fc region.

33. A variant of a parent polypeptide comprising a human IgG Fc region, which variant binds an Fc gamma receptor III (Fcγ

RIII) with better affinity than the parent polypeptide and comprises two or more amino acid substitutions in the Fc region, wherein the Fc region of the variant is not a native sequence Fc region.