Antibody-based therapeutics with enhanced ADCC activity

• US 7,700,321 B2
• Filed: 10/20/2006
• Issued: 04/20/2010
• Est. Priority Date: 10/21/2005
• Status: Active Grant

• Current Assignee: Genzyme Corporation (Sanofi )
• Original Assignee: Genzyme Corporation (Sanofi )
• Inventors: Zhou, Qun, Edmunds, Tim, McPherson, John M.
• Primary Examiner(s): OUSPENSKI, ILIA I
• Application Number:
  US11/551,679
• Publication Number:
  US 20070092521A1
• Time in Patent Office: 1,278 Days
• Field of Search: None
• US Class Current: 435/70.21
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• 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 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• 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 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• 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 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/14 of the thyroid hormones, e.g. T3, T4
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 : Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• 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 16/30 : from tumour cells
• CPC Class Codes: C07K 16/3023 : Lung
• CPC Class Codes: C07K 2317/41 : Glycosylation, sialylation, or fucosylation
• 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

Abstract

Methods for producing antibody-based therapeutics with enhanced ADCC activity are disclosed. The enhanced ADCC activity is attributed to oligomannose-type N-glycans on the antibodies and Fc fusion proteins of the invention. Also disclosed are methods of using such antibody-based therapeutics for targeted killing of cells in a mammal, including therapeutic methods of treating cancers, autoimmune diseases and other diseases.

Claims

1. A method of producing a glycosylated monoclonal antibody or Fc fusion protein comprising:

(a) providing a monoclonal antibody or Fc fusion protein producing cell;

(b) culturing the cell in the presence of an amount of kifunensine sufficient to produce a monoclonal antibody or Fc fusion protein having at least 50% Man_5-9(GlcNAc)₂ N-glycans; and

(c) recovering the glycosylated antibody or Fc fusion protein.

2. The method of claim 1, wherein the amount of kifunensine is less than 1000 ng/ml.

3. The method of claim 1, wherein the amount of kifunensine is less than 500 ng/ml.

4. The method of claim 1, wherein the amount of kifunensine is less than 100 ng/ml.

5. The method of claim 1, wherein the amount of kifunensine is less than 80 ng/ml.

6. The method of claim 1, wherein the amount of kifunensine is at least about 60 ng/ml.

7. The method of claim 1, wherein the amount of kifunensine is about 60 to about 2500 ng/ml.

8. The method of claim 1, wherein the monoclonal antibody or Fc fusion protein has at least 60% Man_5-9(GlcNAc)₂ N-glycans.

9. The method of claim 1, wherein the monoclonal antibody or Fc fusion protein has at least 70% Man_5-9(GlcNAc)₂ N-glycans.

10. The method of claim 1, wherein the monoclonal antibody or Fc fusion protein has at least 90% Man_5-9(GlcNAc)₂ N-glycans.