Covalent diabodies and uses thereof

Patent metadata

Field	Value
Publication number	US-10093739-B2
Application number	US-201615046000-A
Country	US
Kind code	B2
Filing date	Feb 17, 2016
Priority date	Apr 15, 2005
Publication date	Oct 9, 2018
Grant date	Oct 9, 2018

Abstract

The present invention is directed to diabody molecules and uses thereof in the treatment of a variety of diseases and disorders, including immunological disorders, infectious disease, intoxication and cancers. The diabody molecules of the invention comprise two polypeptide chains that associate to form at least two epitope binding sites, which may recognize the same or different epitopes on the same or differing antigens. Additionally, the antigens may be from the same or different molecules. The individual polypeptide chains of the diabody molecule may be covalently bound through non-peptide bond covalent bonds, such as, but not limited to, disulfide bonding of cysteine residues located within each polypeptide chain. In particular embodiments, the diabody molecules of the present invention further comprise an Fc region, which allows antibody-like functionality to engineered into the molecule.

First claim

What is claimed:

1. A method for treating a disease or disorder characterized by a pathogenic antigen, said method comprising administering to a patient in need thereof an effective amount of a diabody molecule, wherein said diabody molecule comprises a first polypeptide chain and a second polypeptide chain, said chains being covalently bonded to one another wherein:
(I) said first polypeptide chain comprises in the N-terminal to C-terminal direction:
(i) a VL1 domain comprising a binding region of a light chain variable domain of a first immunoglobulin specific for a first epitope;
(ii) a VH2 domain comprising a binding region of a heavy chain variable domain of a second immunoglobulin specific for a second epitope; and
(iii) a cysteine-containing hinge domain that comprises:
(a) the amino acid sequence: VEPKSC (SEQ ID NO:79); or
(b) the amino acid sequence: AEPKSC (amino acid residues 245-250 of SEQ ID NO:18);

 wherein said VL1 domain and said VH2 domain are covalently linked by a peptide linker such that said VL1 domain and said VH2domain do not associate with one another to form an epitope binding site for said first or second epitope;
(II) said second polypeptide chain comprises in the N-terminus to C-terminus direction:
(i) a VL2 domain comprising a binding region of a light chain variable domain of the second immunoglobulin;
(ii) a VH1 domain comprising a binding region of a heavy chain variable domain of the first immunoglobulin; and
(iii) a cysteine-containing light chain constant domain that comprises the C-terminal 2 to 8 amino acid residues of a human light chain constant domain;

 wherein said VL2 domain and said VH1 domain are covalently linked by a peptide linker such that said VL2 domain and said VH1 domain do not associate with one another to form an epitope binding site for said first or second epitope; and
(III) (i) said VL1 domain and said VH1 domain associate with one another to form a first binding site (VL1)(VH1) that binds said first epitope;
(ii) said VH2 domain and said VL2 domain associate with one another to form a second binding site (VL2)(VH2) that binds said second epitope;
(iii) said first and second epitopes are different; and
(iv) said first polypeptide chain and said second polypeptide chain are covalently linked via a disulfide bond between said cysteine residue of said hinge domain and said cysteine of said light chain constant domain;

and wherein one of said first and said second binding sites binds to said pathogenic antigen.

2. The method of claim 1 , wherein said cysteine-containing hinge domain additionally comprises an Fc domain or portion thereof.

3. The method of claim 1 , wherein said light chain constant domain is a domain of a human kappa light chain or a human lambda light chain.

4. The method of claim 1 , wherein one of said first and said second binding sites binds to said pathogenic antigen and the second of said first and said second binding sites binds to an antigen expressed on the surface of a B-cell, a T-cell, a phagocytic cell, a natural killer (NK) cell or a dendritic cell.

5. The method of claim 1 , wherein said disease or disorder is cancer and said pathogenic antigen is a cancer antigen that is characteristic of said cancer.

6. The method of claim 5 , wherein said cancer is a bladder cancer, a breast cancer, a cancer of mesenchymal origin, a cancer of the central and peripheral nervous system, a cervical cancer, a colon cancer, a hematopoietic cancer, a kidney cancer, a liver cancer, a lung cancer, a pancreatic cancer, a prostate cancer, a skin cancer, a stomach cancer, a thyroid cancer, or an ovarian cancer.

7. The method of claim 1 , wherein said disease or disorder is an infectious disease and said pathogenic antigen is a bacterial antigen, a viral antigen, a parasite antigen, or a fungal antigen that is characteristic of said infectious disease.

8. The method of claim 1 , wherein said disease or disorder is an autoimmune disease, wherein one of said first and said second binding sites binds to a pathogenic antigen that is an autoimmune antigen characteristic of said autoimmune disease, and the other of said first and said second binding sites binds to a FcγRIIB.

9. The method of claim 8 , wherein said autoimmune disease is selected from the group consisting of: alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, autoimmune disease of the adrenal gland, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, Behcet's disease, bullous pemphigoid, cardiomyopathy, celiac sprue-dermatitis, chronic fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, Churg-Strauss syndrome, cicatrical pemphigoid, CREST syndrome, cold agglutinin disease, Crohn's disease, discoid lupus, essential mixed cryoglobulinemia, fibromyalgia-fibromyositis, glomerulonephritis, Graves' disease, Guillain-Barre, Hashimoto's thyroiditis, idiopathic pulmonary fibrosis, idiopathic thrombocytopenia purpura (ITP), IgA neuropathy, juvenile arthritis, lichen planus, lupus erthematosus, Ménière's disease, mixed connective tissue disease, multiple sclerosis, type 1 or immune-mediated diabetes mellitus, myasthenia gravis, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychrondritis, polyglandular syndrome, polymyalgia rheumatica, polymyositis and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynauld's phenomenon, Reiter's syndrome, Rheumatoid arthritis, sarcoidosis, scleroderma, Sjögren' s syndrome, stiff-man syndrome, systemic lupus erythematosus, lupus erythematosus, takayasu arteritis, temporal arteristis/giant cell arteritis, ulcerative colitis, uveitis, and a vasculitide.

10. The method of claim 8 , wherein said autoimmune disease is an inflammatory disorder selected from the group consisting of: asthma, encephilitis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), an allergic disorder, septic shock, pulmonary fibrosis, undifferentitated spondyloarthropathy, undifferentiated arthropathy, arthritis, inflammatory osteolysis, and a chronic inflammation resulting from chronic viral or bacterial infection.

11. The method of claim 8 , wherein said autoimmune antigen is selected from the group consisting of DNA, RNA and collagen.

12. The method of claim 3 , wherein said light chain constant domain comprises the amino acid sequence FNRGEC (SEQ ID NO:23).

13. The method of claim 9 , wherein said vasculitide is dermatitis herpetiformis vasculitis, vitiligo, or Wegener's granulomatosis.

Assignees

• Macrogenics Inc

Inventors

• Johnson Leslie S
• Huang Ling

Classifications

• A61K2039/505

  comprising antibodies · CPC title

• C07K16/12

  against material from bacteria · CPC title

• C07K2317/52

  Constant or Fc region; Isotype · CPC title

• C07K2317/53

  Hinge · CPC title

• A61P31/00

  Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics · CPC title