Engineered immunoglobulin FC polypeptides

• US 8,952,132 B2
• Filed: 02/06/2012
• Issued: 02/10/2015
• Est. Priority Date: 02/07/2011
• Status: Active Grant

• Current Assignee: Research & Development Foundation
• Original Assignee: Research & Development Foundation
• Inventors: Georgiou, George, Jung, Sang Taek, Kelton, William, Kang, Tae Hyun
• Primary Examiner(s): DAHLE, CHUN WU
• Application Number:
  US13/367,063
• Publication Number:
  US 20120208238A1
• Time in Patent Office: 1,100 Days
• Field of Search: None
• US Class Current: 530/350
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• 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 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 16/283 against Fc-receptors, e.g. CD16, CD32, CD64 CD23 C07K16/2851
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 2317/14 Specific host cells or culture conditions, e.g. components, pH or temperature
• CPC Class Codes: C07K 2317/41 Glycosylation, sialylation, or fucosylation
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• 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 2317/92 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

Abstract

Methods and compositions involving polypeptides having an aglycosylated antibody Fc domain. In certain embodiments, polypeptides have an aglycosylated Fc domain that contains one or more substitutions compared to a native Fc domain. Additionally, some embodiments involve an Fc domain that is binds some Fc receptors but not others. For example, polypeptides are provided with an aglycosylated Fc domain that selectively binds FcγRIIa, but that is significantly reduced for binding to the highly homologous FcγRIIb receptors. Furthermore, methods and compositions are provided for promoting antibody-dependent cell-mediated toxicity (ADCC) using a polypeptide having a modified aglycosylated Fc domain and a second non-Fc binding domain, which can be an antigen binding region of an antibody or a non-antigen binding region. Some embodiments concern antibodies with such polypeptides, which may have the same or different non-Fc binding domain.

Claims

1. A polypeptide comprising an aglycosylated antibody human variant IgG Fc domain capable of binding a human Fcγ

RIIa polypeptide, wherein the Fc domain comprises an S298G and T299A amino acid substitutions at amino acids 298 and 299, wherein the numbering of the residues in the Fc domain is that of the EU index as in Kabat, and further wherein the variant IgG Fc domain comprises at least one additional substitution selected from the group consisting of;

a) E382V and V263E;

b) E382V, N390D and M428L;

c) K392E, E382V, V397M and M428L;

d) N315D, E382V and M428L, or e) H268P, E294K, N361S, E382V and M428L.

2. The polypeptide of claim 1, wherein the Fc domain comprises the substitutions S298G, T299A, E382V and V263E.

3. The polypeptide of claim 2, wherein the Fc domain comprises SEQ ID NO:

9.

4. The polypeptide of claim 1, wherein the Fc domain comprises the substitutions S298G, T299A, E382V, N390D and M428L.

5. The polypeptide of claim 4, wherein the Fc domain comprises SEQ ID NO:

7.

6. The polypeptide of claim 1, wherein the Fc domain comprises the substitutions S298G, T299A, E382V, K392E, V397M and M428L.

7. The polypeptide of claim 6, wherein the Fc domain comprises SEQ ID NO:

6.

8. The polypeptide of claim 1, wherein the Fc domain comprises the substitutions S298G, T299A, N315D, E382V, and M428L.

9. The polypeptide of claim 8, wherein the Fc domain comprises SEQ ID NO:

5.

10. The polypeptide of claim 1, wherein the Fc domain comprises the substitutions S298G, T299A, H268P, E294K, N361S, E382V and M428L.

11. The polypeptide of claim 10, wherein the Fc domain comprises SEQ ID NO:

4.

12. The polypeptide of claim 1, further comprising a non-FcR binding domain.

13. The polypeptide of claim 12, wherein the non-FcR binding domain is an antigen binding site of an antibody.

14. The polypeptide of claim 12, wherein the non-FcR binding region is not an antigen binding site of an antibody.

15. The polypeptide of claim 14, wherein the non-FcR binding region binds a cell-surface protein.

16. The polypeptide of claim 15, wherein the cell-surface protein is a receptor.

17. The polypeptide of claim 16, wherein the receptor is a tyrosine kinase.

18. The polypeptide of claim 17, wherein the non-FcR binding region binds multiple tyrosine kinase receptors.