ENGINEERED IMMUNOGLOBULIN FC POLYPEPTIDES DISPLAYING IMPROVED COMPLEMENT ACTIVATION

• US 20160297886A1
• Filed: 02/09/2016
• Published: 10/13/2016
• Est. Priority Date: 02/09/2015
• Status: Active Grant

• Current Assignee: Research & Development Foundation
• Original Assignee: Research & Development Foundation
• Inventors: LEE, Chang-Han, GEORGIOU, George
• Granted Patent: US 10,457,737 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• 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 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/2887 against CD20
• 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 2317/524 CH2 domain
• CPC Class Codes: C07K 2317/72 Increased 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 2317/77 Internalization into the cell
• CPC Class Codes: C07K 2317/90 characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2317/94 Stability, e.g. half-life, pH, temperature or enzyme-resistance

Abstract

Methods and compositions involving polypeptides having an aglycosylated antibody Fc domain are provided. 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 Clq, and optionally activating Fc receptors, but that is significantly reduced for binding to the inhibitory FcγRIIb receptor. Furthermore, methods and compositions are provided for promoting complement dependent cytotoxicity (CDC) 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 a different non-Fc binding domain.

Claims

1. A polypeptide comprising an aglycosylated variant human IgG Fc domain capable of binding human Clq, wherein the Fc domain comprises an amino acid substitution selected from the group consisting of:

(a) 308, 337, 338, 340, 342, 344, 345, and 372;

(b) 320 and 386;

(c) 235, 236, 237, and 351;

(d) 246, 322, and 402;

(e) 242, 315, 336, 340, 342, 378, and 386;

(f) 334, 351, and 421;

(g) 341 and 351;

(h) 252, 341, and 351;

(i) 246, 260, 315, and 386;

(j) 246, 252, 322, 344, 345, and 372;

(k) 242, 252, 338, 341, and 345; and

(1) 334, 402, 338, 342, 344, 345, and 372, wherein the numbering of the residues in the Fc domain is that of the EU index as in Kabat.

2. The polypeptide of claim 1, wherein the polypeptide has specific affinity for binding human Clq.

3. The polypeptide of claim 1, wherein the polypeptide does not detectably bind Fcγ

receptors.

4. The polypeptide of claim 1, wherein the polypeptide has specific affinity for binding human Clq and activating Fc receptors.

5. The polypeptide of claim 4, wherein the polypeptide does not detectably bind Fcγ

RIIb.

6. The polypeptide of claim 2, wherein the aglycosylated variant human IgG Fc domain comprises substitutions K320E and Q386R.

7. The polypeptide of claim 6, wherein the aglycosylated variant human IgG Fc domain is Fc801 (SEQ ID NO:

7).

8. The polypeptide of claim 2, wherein the aglycosylated variant human IgG Fc domain comprises substitutions L235K, G236M, G237R, and L351Q.

9. The polypeptide of claim 8, wherein the aglycosylated variant human IgG Fc domain is Fc802 (SEQ ID NO:

10).

10. The polypeptide of claim 4, wherein the aglycosylated variant human IgG Fc domain comprises substitutions V308A, S337P, K338Q, K340R, Q342P, R344G, E345Y, and F372L.

11. The polypeptide of claim 10, wherein the aglycosylated variant human IgG Fc domain is Fc805 (SEQ ID NO:

22).

12. The polypeptide of claim 4, wherein the aglycosylated variant human IgG Fc domain comprises the substitution M252V.

13. The polypeptide of claim 12, wherein the aglycosylated variant human IgG Fc domain is Fc-V1 (SEQ ID NO:

36).

14. The polypeptide of claim 12, wherein the aglycosylated variant human IgG Fc domain further comprises the substitutions G431A and L351Q.

15. The polypeptide of claim 14, wherein the aglycosylated variant human IgG Fc domain is Fc-V18 (SEQ ID NO:

41).

16. The polypeptide of claim 12, wherein the aglycosylated variant human IgG Fc domain further comprises the substitutions K246N, K322E, R344G, E345Y, and F372L.

17. The polypeptide of claim 16, wherein the aglycosylated variant human IgG Fc domain is Fc-V23 (SEQ ID NO:

43).

18. The polypeptide of claim 12, wherein the aglycosylated variant human IgG Fc domain further comprises the substitutions F242L, K338Q, G341A, and E345Y.

19. The polypeptide of claim 18, wherein the aglycosylated variant human IgG Fc domain is Fc-V24 (SEQ ID NO:

44).

53. A method of treating a subject having a tumor comprising administering to the subject an effective amount of a polypeptide according to claim 1 in a pharmaceutically acceptable carrier.

20-52. -52. (canceled)

54-66. -66. (canceled)