Identification and engineering of antibodies with variant Fc regions and methods of using same

• US 7,355,008 B2
• Filed: 01/09/2004
• Issued: 04/08/2008
• Est. Priority Date: 01/09/2003
• Status: Active Grant

• Current Assignee: MacroGenics, Inc., Marcogenics Inc.
• Original Assignee: MacroGenics, Inc.
• Inventors: Stavenhagen, Jeffrey, Huang, Ling, Rankin, Christopher, Gorlatov, Sergey, Vijh, Sujata
• Primary Examiner(s): Haddad; Maher M.
• Assistant Examiner(s): DAHLE, CHUN WU
• Application Number:
  US10/754,922
• Publication Number:
  US 20050037000A1
• Time in Patent Office: 1,551 Days
• Field of Search: None
• US Class Current: 530/387.1
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 11/02 Nasal agents, e.g. decongestants
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/02 of urine or of the urinary tract, e.g. urine acidifiers
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• 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 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/20 for DNA viruses
• CPC Class Codes: A61P 33/00 Antiparasitic agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 37/08 : Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/36 : from Actinomyces; from Streptomyces (G)
• CPC Class Codes: C07K 14/405 : from algae
• CPC Class Codes: C07K 14/70535 : Fc-receptors, e.g. CD16, CD32, CD64 (CD2314/705F)
• CPC Class Codes: C07K 16/005 : constructed by phage libraries
• CPC Class Codes: C07K 16/283 : against Fc-receptors, e.g. CD16, CD32, CD64 CD23 C07K16/2851
• CPC Class Codes: C07K 16/2887 : against CD20
• CPC Class Codes: C07K 16/32 : against translation products of oncogenes
• CPC Class Codes: C07K 16/44 : against material not provided for elsewhere , e.g. haptens, metals, DNA, RNA, amino acids
• CPC Class Codes: C07K 2317/24 : containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/52 : Constant or Fc region; Isotype
• 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 2317/76 : Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2319/70 : containing domain for protein-protein interaction
• CPC Class Codes: G01N 2333/70535 : Fc-receptors, e.g. CD16, CD32, CD64 (CD2314/705F)
• CPC Class Codes: G01N 33/6857 : Antibody fragments

Abstract

The present invention relates to molecules, particularly polypeptides, more particularly immunoglobulins (e.g., antibodies), comprising a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification relative to a wild-type Fc region, which variant Fc region binds FcγRIIA and/or FcγRIIA with a greater affinity, relative to a comparable molecule comprising the wild-type Fc region. The molecules of the invention are particularly useful in preventing, treating, or ameliorating one or more symptoms associated with a disease, disorder, or infection. The molecules of the invention are particularly useful for the treatment or prevention of a disease or disorder where an enhanced efficacy of effector cell function (e.g., ADCC) mediated by FcγR is desired, e.g., cancer, infectious disease, and in enhancing the therapeutic efficacy of therapeutic antibodies the effect of which is mediated by ADCC.

Claims

1. An antibody which comprises an antigen binding region and a variant Fe region, wherein said variant Fc region:

(A) differs from a wild-type Fe region by comprising an amino acid modification at position 396, according to the EU index as in Kabat, relative to said wild-type Fc region; and

(B) binds an Fcγ

R with an increased affinity relative to a said wild-type Fc region.

2. The antibody of claim 1, wherein said variant Fc region differs from said wild-type Fc region in comprising additional amino acid modification(s) at position 210;

215;

217;

246;

250;

255;

268;

288;

290;

or 419 of said wild-type Fc region.

3. The antibody of claim 1, wherein said variant Fc region differs from said wild-type Fc region in comprising additional amino acid modification(s) of said wild-type Fc region selected from the group consisting of:

210GM;

217S;

227S;

240A;

242F;

244H;

246T;

247S;

248M;

250A;

250S;

255I;

255L;

258D;

268D;

268N;

303I;

305L;

323I;

326I;

334N;

358P;

370E;

375C;

379M;

384K;

392T;

400F;

410H;

419H;

419L;

or 427A.

4. The antibody of claim 1, wherein said variant Fc region differs from said wild-type Fc region in comprising additional amino acid modification(s) of said wild-type Fc region selected from the group consisting of:

(1) 221E;

270E;

308A, 311H and 402D;

(2) 319F and 352L;

(3) 288R;

307A; and

344E;

(4) 210M and 261N; and

(5) 243L;

305I;

378D; and

404S.

5. The antibody of any of claims 1-4, wherein said Fcγ

R is Fcγ

RIIIA.

6. The antibody of claim 5, wherein said variant Fc region of said antibody has decreased affinity for Fcγ

RIIB relative to said wild-type Fc region.

7. The antibody of claim 6, wherein said variant Fc region comprises amino acid modifications of said wild-type Fc region selected from the group consisting of:

(1) 221E;

270E;

308A;

311H;

396L and 402D;

(2) 243L;

305I;

376D;

404S; and

396L;

(3) 255I and 396L;

(4) 370E and 396L.

(5) 392T and 396L; and

(6) 410H and 396L.

8. The antibody of any of claims 1-4 wherein said wild-type Fc region is an Fc region of a human IgG Fc region.

9. The antibody of claim 8, wherein said human IgG Fe region is a human IgG1, IgG2, IgG3, or IgG4 Fc region.

10. The antibody of claim 8, wherein said variant Fc region of said antibody specifically binds Fcγ

RIIIA with at least two times greater affinity than the wild-type Fc region.

11. The antibody of claim 1-4, wherein said antigen binding region binds a cancer antigen or an antigen associated with an infectious disease.

12. The antibody of claim 11, wherein said antibody binds a cancer antigen and mediates antibody dependent cell mediated cytotoxicity 2-fold more effectively than an antibody comprising said wild-type Fc region.

13. The antibody of claim 11, wherein said antibody binds a cancer antigen and said cancer antigen is HER-2/neu, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, MUM-1, CDK4, MUC-1, N-acetylglucosaminyltransferase, p15, beta-catenin, human papillomavirus-E6, or human papillomavirus-E7.

14. The antibody of claim 1, wherein said amino acid modification at position 396 is a substitution of leucine at said position.

15. The antibody of claim 5, wherein said amino acid modification at position 396 is a substitution of leucine at said position.

16. The antibody of claim 6, wherein said amino acid modification at position 396 is a substitution of leucine at said position.

17. The antibody of claim 8, wherein said amino acid modification at position 396 is a substitution of leucine at said position.

18. The antibody of claim 10, wherein said amino acid modification at position 396 is a substitution of leucine at said position.

19. The antibody of claim 11, wherein said amino acid modification at position 396 is a substitution of leucine at said position.

20. The antibody of claim 12, wherein said amino acid modification at position 396 is a substitution of leucine at said position.

21. The antibody of claim 13, wherein said amino acid modification at position 396 is a substitution of leucine at said position.

22. The antibody of claim 14, wherein said amino acid modification at position 396 is a substitution of leucine at said position.