Polypeptide variants

• US 6,528,624 B1
• Filed: 03/31/1999
• Issued: 03/04/2003
• Est. Priority Date: 04/02/1998
• Status: Expired due to Term

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Mulkerrin, Michael George, Idusogie, Esohe Ekinaduese, Presta, Leonard G.
• Primary Examiner(s): Nolan, Patrick J.
• Application Number:
  US09/282,846
• Time in Patent Office: 1,434 Days
• Field of Search: 435/328, 424/142.1, 530/387.3, 530/387.1
• US Class Current: 530/387.3
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: A61K 47/6873 the antibody targeting an immunoglobulin; the antibody being an anti-idiotypic antibody
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 16/4291 against IgE
• 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 2317/71 Decreased effector function due to an Fc-modification
• CPC Class Codes: C07K 2317/72 Increased 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 2319/00 Fusion polypeptide

Abstract

A variant of a polypeptide comprising a human IgG Fc region is described, which variant comprises an amino acid substitution at one or more of amino acid positions 270, 322, 326, 327, 329, 331, 333 or 334 of the human IgG Fc region. Such variants display altered effector function. For example, C1q binding and/or complement dependent cytotoxicity (CDC) activity may be altered in the variant polypeptide. The application also discloses a variant of a parent polypeptide comprising a human IgG Fc region, which variant has a better binding affinity for human C1q than the parent polypeptide.

Claims

1. A variant of an antibody or immunoadhesin comprising a human IgG Fec region, which variant comprises an amino acid substitution at amino acid position 270 or 329, or at two or more of amino acid positions 270, 322, 329 and 331 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat, wherein the antibody variant is able to bind an antigen, and the immunoadhesin variant is able to bind a ligand or receptor.

2. The variant of claim 1 having Pro329 of the human IgG Fc region substituted with another amino acid.

3. The variant of claim 1 having Asp270 of the human IgG Fc region substituted with another amino acid.

4. The variant of claim 1 wherein the polypeptide comprises an antibody.

5. The variant of claim 1 wherein the polypeptide comprises a human IgG1 Fc region.

6. The variant of claim 1 which does not activate complement.

7. The variant of claim 6 which binds an FcR.

8. The variant of claim 7 which binds Fcγ

RI, Fcγ

RII, Fcγ

RIII or FcRn.

10. A composition comprising the variant of claim 1 and a physiologically acceptable carrier.

9. A variant of an antibody or immunoadhesin comprising a human IgG Fc region, which variant binds Fcγ

RI, Fcγ

RII, Fcγ

RIII or FcRn but does not activate complement and comprises an amino acid substitution at amino acid position 322 or amino acid position 329, or both amino acid positions of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat, wherein the antibody variant is able to bind an antigen, and the immunoadhesin variant is able to bind a ligand or receptor.

11. A method for modifying an antibody or irmunoadhesin comprising a human IgG Fc region, the method comprising substituting an amino acid residue at amino acid position 270 or 329, or at two or more of amino acid positions 270, 322, 329 and 331 of the human IgG Of region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat, to generate a variant wherein the antibody variant is able to bind an antigen, and the immunoadhesin variant is able to bind a ligand or receptor.

12. A variant of an antibody or immunoadhesin comprising a human IgG Fc region, which variant comprises an amino acid substitution at amino acid position 326, 327, 333 or 334 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat, wherein the antibody variant is able to bind an antigen, and the immunoadhesin variant is able to bind a ligand or receptor.

13. The variant of claim 12 which comprises amino acid substitutions at two or more of amino acid positions 326, 327, 333 or 334.

14. The variant of claim 13 which comprises amino acid substitutions at three or more amino acid positions 326, 327, 333 or 334.

15. The variant of claim 14 which comprises amino acid substitutions at amino acid positions 326, 327, 333 and 334.

16. A method for modifying an antibody or immunoadhesin comprising a human IgG Fc region, the method comprising substituting an amino acid residue at amino acid position 326, 327, 333 or 334 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat, to generate a variant wherein the antibody variant is able to bind an antigen, and the immunoadhesin variant is able to bind a ligand or receptor.

17. A variant of an antibody comprising a human IgG Fc region, which variant comprises an amino acid substitution at amiao acid position 270 or 329, or at two or more of amino acid positions 270, 322, 329 and 331 of the human IgG Fc region, where the numbering of the residues in the IgG Fec region is that of the EU index as in Kabat, wherein the antibody variant binds an antigen.

18. A variant of an antibody comprising a human TgG Fc region, which variant comprises an amino acid substitution at amino acid position 326, 327, 333 or 334 of the human IgG Fc region, where the numbering of the residues in the IgG Fc region is that of the EU index as in Kabat, wherein the antibody variant binds an antigen.

19. The variant of claim 18 wherein the antigen is CD20.

20. The variant of claim 18 wherein the antigen is HER2 receptor.