ANTIBODY Fc VARIANTS

• US 20120251531A1
• Filed: 03/27/2012
• Published: 10/04/2012
• Est. Priority Date: 03/29/2011
• Status: Active Grant

• Current Assignee: Roche Glycart AG (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Baehner, Monika, Kubbies, Manfred, Moessner, Ekkehard, Schlothauer, Tilman, Jenewein, Stefan
• Granted Patent: US 8,969,526 B2
• Time in Patent Office: Days
• US Class Current: 424/133.1
• 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 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• 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 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/71 Decreased effector function due to an Fc-modification
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• 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 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: Y10S 435/81 Packaged device or kit

Abstract

The invention relates to engineered polypeptides comprising Fc variants and their uses. More specifically, Fc variants are described exhibiting reduced effector function. These variants cause a benefit for a patient suffering from a disease which could be treated with an antibody for which it is desirable to reduce the effector function elicited by antibodies.

Claims

1. A polypeptide comprising an Fc variant of a wild-type human IgG Fc region, said Fc variant comprising an amino acid substitution at position Pro329 and at least one further amino acid substitution, wherein the residues are numbered according to the EU index of Kabat, and wherein said polypeptide exhibits a reduced affinity to one or more of human Fcγ

RIIIA, Fcγ

RIIA, and Fcγ

RI compared to the polypeptide comprising the wildtype IgG Fc region, and wherein the antibody-dependent cell-mediated cytotoxicity (ADCC) induced by said polypeptide is reduced to at least 20% of the ADCC induced by the polypeptide comprising a wild-type human IgG Fc region.

2. The polypeptide according to claim 1, wherein Pro329 of the wild-type human Fc region is substituted with glycine or arginine or an amino acid residue large enough to destroy the proline sandwich within the Fc region.

3. The polypeptide according to claim 1, wherein said at least one further amino acid substitution is selected from S228P, E233P, L234A, L235A, L235E, N297A, N297D, and P331S.

4. The polypeptide according to claim 3, wherein said at least one further amino acid substitution is L234A and L235A of the human IgG1 Fc region or S228P and L235E of the human IgG4 Fc region.

5. The polypeptide according to claim 1, wherein the affinity to at least one further human receptor reduced compared to the polypeptide comprising a wild-type human IgG Fc region wherein the further human receptor is selected from Fcγ

I, Fcγ

IIA, and C1q.

6. The polypeptide according to claim 1, wherein the polypeptide comprises a human IgG1 or IgG4 Fc region.

7. The polypeptide according to claim 1, wherein the polypeptide is an antibody or an Fc fusion protein.

8. The polypeptide according to claim 1, wherein thrombocyte aggregation induced by the polypeptide is reduced compared to the thrombocyte aggregation induced by a polypeptide comprising a wild-type human IgG Fc region.

9. The polypeptide according to claim 1, wherein CDC induced by the polypeptide is strongly reduced compared to the CDC induced by a polypeptide comprising a wild-type human IgG Fc region.

11. The polypeptide according to any one of claims 1-9, wherein the polypeptide is an anti-CD9 antibody comprising a heavy chain variable region comprising SEQ ID NO:

9 and a variable light chain region comprising SEQ ID NO;

8.

12. A method of treating a disease comprising administering the polypeptide according to claim 20, wherein it is favorable for treating the disease that an effector function of the polypeptide is strongly reduced compared to the effector function induced by the polypeptide comprising a wild-type human IgG Fc region.

14. A method of treating an individual having a disease, wherein it is favorable that the effector function of the polypeptide comprising an Fc variant of a wild-type human IgG Fc region is strongly reduced compared to the effector function induced by the polypeptide comprising a wildtype human Fc polypeptide, comprising administering to an individual an effective amount of the polypeptide according to claim 20.

20. A pharmaceutical formulation comprising the polypeptide according to claim 11.

21. An article of manufacture comprising the polypeptide according to claim 11.

10. (canceled)

13. (canceled)

15. A method for down-modulating ADCC comprising a polypeptide comprising an Fc variant of a wild-type human IgG Fc region, said polypeptide having Pro329 of the human IgG Fc region substituted with glycine, wherein the residues are numbered according to the EU index of Kabat, wherein said polypeptide exhibits a reduced affinity to the human Fcγ

RIIIA and Fcγ

RIIA, wherein ADCC is down-modulated to at least 20% of the ADCC induced by the polypeptide comprising the wildtype human IgG Fc region.

16. The method according to any one of claims 15, 22, or 23 wherein the Fc variant comprises at least two further amino acid substitution at L234A and L235A of the human IgG1 Fc region or S228P and L235E of the human IgG4 Fc region.

17. The method of claim 16, wherein thrombocyte aggregation induced by the polypeptide is reduced compared to the thrombocyte aggregation induced by the polypeptide comprising a wildtype human Fc region, wherein the polypeptide is a platelet activating antibody.

22. The method of claim 15 further comprising down-modulating antibody-dependent cellular phagocytosis (ADCP).

18. A method of treating an individual having a disease with a polypeptide, said polypeptide having Pro329 of the human IgG Fc region substituted with Gly, wherein the residues are numbered according to the EU index of Kabat, wherein said polypeptide is characterized by a strongly reduced binding to Fcγ

RIIIA and Fcγ

RIIA compared to the polypeptide comprising a wildtype human IgG Fc region, comprising administering to the individual an effective amount of said polypeptide.

19. The method according to claim 18, wherein said polypeptide comprises at least two further amino acid substitution at L234A and L235A of the human IgG1 Fc region or S228P and L235E of the human IgG4 Fc region.

23. A method for down-modulating ADCP comprising a polypeptide comprising an Fc variant of a wild-type human IgG Fc region, said polypeptide having Pro329 of the human IgG Fc region substituted with glycine, wherein the residues are numbered according to the EU index of Kabat, wherein said polypeptide exhibits a reduced affinity to the human Fcγ

RIIIA and Fcγ

RIIA.