Covalently-associated diabody complexes that possess charged coil domains and that are capable of enhanced binding to serum albumin

Patent metadata

Field	Value
Publication number	US-9889197-B2
Application number	US-201113811883-A
Country	US
Kind code	B2
Filing date	Jul 29, 2011
Priority date	Apr 15, 2005
Publication date	Feb 13, 2018
Grant date	Feb 13, 2018

Abstract

Diabody molecules and uses thereof in the treatment of a variety of diseases and disorders, including immunological disorders, infectious disease, intoxication and cancers are disclosed. The diabody molecules 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 disulfide bonding of cysteine residues located within each polypeptide chain. The diabody molecules may further comprise an Fc region, which allows antibody-like functionality to be engineered into the molecule.

First claim

What is claimed is:

1. A diabody complex that consists of a first polypeptide chain, a second polypeptide chain and a third polypeptide chain, each comprising an N-terminus and a C-terminus, wherein:
(A) said first polypeptide chain of said diabody complex comprises, in the N-terminal to C-terminal direction:
(1) a first domain comprising a binding region of a light chain variable domain of a first immunoglobulin, said first immunoglobulin being specific for a first epitope;
(2) a peptide linker having fewer than 12 amino acid residues; and
(3) a second domain comprising a binding region of a heavy chain variable domain of a second immunoglobulin, said second immunoglobulin being specific for a second epitope; and

(B) said second polypeptide chain of said diabody complex comprises, in the N-terminal to C-terminal direction:
(1) a first domain comprising a binding region of a light chain variable domain of the second immunoglobulin;
(2) a peptide linker having fewer than 12 amino acid residues; and
(3) a second domain comprising a binding region of a heavy chain variable domain of the first immunoglobulin; and

(C) said third polypeptide chain of said diabody complex comprises:
(1) a polypeptide portion of a protein that binds to serum albumin, said polypeptide portion being capable of binding to said serum albumin, and
(2) a coil domain that is an E-coil domain comprising the amino acid sequence of SEQ ID NO:299 or a K-coil domain comprising the amino acid sequence of SEQ ID NO:300;

wherein:
(1) said first domain of said first polypeptide chain of said diabody complex comprises three light chain CDRs and said second domain of said second polypeptide chain of said diabody complex comprises three heavy chain CDRs, and wherein said domains associate to form a first binding site that binds the first epitope;
(2) said first domain of said second polypeptide chain of said diabody complex comprises three light chain CDRs and said second domain of said first polypeptide chain of said diabody complex comprises three heavy chain CDRs and wherein said domains associate to form a second binding site that binds the second epitope;
(3) one of said first or said second polypeptide chains of said diabody complex additionally comprises a C-terminally positioned coil domain that is an E-coil domain comprising the amino acid sequence of SEQ ID NO:299 or a K-coil domain comprising the amino acid sequence of SEQ ID NO:300;
(4) when said coil domain of said first or second polypeptide chain of said diabody complex is an E-coil, said coil domain of said third polypeptide chain of said diabody complex is a K-coil, and when said coil domain of said first or second polypeptide chain of said diabody complex is a K-coil, said coil domain of said third polypeptide chain of said diabody complex is an E-coil, wherein said coil domain of said first or second polypeptide chain of said diabody complex associates with said coil domain of said third polypeptide chain of said diabody complex.

2. The diabody complex of claim 1 , wherein said serum albumin binding protein is streptococcal protein G and said polypeptide portion of said serum albumin-binding protein is an albumin-binding domain (ABD) of said streptococcal protein G.

3. The diabody complex of claim 2 , wherein said albumin-binding domain (ABD) of said streptococcal protein G is albumin-binding domain 3 (ABD3) of protein G of Streptococcus strain G148.

4. The diabody complex of claim 1 , wherein said diabody complex exhibits an in vivo serum half-life greater than 2 hours.

5. The diabody complex of claim 1 , wherein said first or second epitope is an epitope from a Natural Killer Group 2D (NKG2D) receptor, an epitope from a T cell receptor (TCR), an epitope from CD3, or an epitope from a hapten.

6. The diabody complex of claim 5 , wherein said diabody of said complex additionally binds to a tumor-associated antigen.

7. The diabody complex of claim 6 , wherein said tumor-associated antigen is a breast cancer antigen, an ovarian cancer antigen, a prostate cancer antigen, a cervical cancer antigen, a pancreatic carcinoma antigen, a lung cancer antigen, a bladder cancer antigen, a colon cancer antigen, a testicular cancer antigen, a glioblastoma cancer antigen, an antigen associated with a B cell malignancy, an antigen associated with multiple myeloma, an antigen associated with non-Hodgkin's lymphoma, or an antigen associated with chronic lymphocytic leukemia.

8. The diabody complex of claim 7 , wherein said tumor-associated antigen is colorectal carcinoma antigen (A33 antigen); Disintegrin and Metalloproteinase Domain-Containing Protein 9 (ADAM-9); CD166 Antigen (ALCAM); Cyclin B1 (B1 antigen); B Melanoma Antigen (BAGE); beta-catenin; Cancer Antigen 125 (CA125); Carboxypeptidase M; CD5 Antigen (CD5); B-Lymphocyte Antigen CD19 (CD19); B-Lymphocyte Antigen CD20 (CD20); Cluster of Differentiation 22 (CD22); Fc Epsilon RH (CD23); IL-2 Receptor Alpha Chain (CD25); CD27 Antigen (CD27); Cluster of Differentiation 28 (CD28); Low Affinity Immunoglobulin Gamma Fc Receptor II-b (CD32B); Cluster of Differentiation 36 (CD36); Cluster of Differentiation 40 (CD40); Cluster of Differentiation 45 (CD45); Cluster of Differentiation 46 (CD46); Neural cell adhesion molecule (CD56); B-cell antigen Receptor Complex-Associated Protein Alpha Chain (CD79a); CD79B antigen like complex (CD79bl); Cluster of Differentiation 103 (CD103); CD40 Ligand (CD154); Cyclin-Dependent Kinase 4 (CDK4); Carcinoembryonic Antigen (CEA); Cytotoxic T-Lymphocyte-Associated Protein 4 (CTLA4); Cytokeratin 8; an Ephrin receptor; Epidermal Growth Factor Receptor (ErbB1); Erb-B2 Receptor Tyrosine-Protein Kinase-3 (ErbB3); Erb-B2 Receptor Tyrosine-Protein Kinase-4 (ErbB4); G Antigen 1 (GAGE-1); G Antigen 2 (GAGE-2); Ganglioside G2 (GD2); Ganglioside G3 (GD3); Ganglioside GM2 (GM2); Melanoma Differentiation Antigen (gp100); Receptor Tyrosine-Protein Kinase erbB-2 (HER-2/neu); human papillomavirus-E6; human papillomavirus-E7; Integrin Alpha-V-Beta-6; Junctional Adhesion Molecule C (JAM-3); KID3 Antigen; KID31 Antigen; Colorectal Carcinoma-Associated GA733 Antigen; Epithelial Cell Adhesion Molecule (KSA); LUCA-2 Antigen; Melanoma-Associated Antigen 1 (MAGE-1); Melanoma-Associated Antigen 3 (MAGE-3); Melanoma-Associated Antigen Recognized by T Cells (MART); Mucin 1 Cell Surface Associated (MUC-1); Mutated Melanoma-Associated Antigen 1 (MUM-1); N-acetylglucosaminyltransferase; Oncostatin M (Oncostatin Receptor Beta); Cyclin-Dependent Kinase Inhibitor 2B (p15); PIPA Antigen; Prostate-Specific Antigen (P SA); Prostate-Specific Membrane Antigen (PSMA); RAAG10 Antigen; Receptor Tyrosine Kinase-Like Orphan Receptor 1 (ROR1); Sialyl-Tn (sTn); TEST Antigen; Tumor Necrosis Factor-Alpha Receptor (TNF-α receptor); Tumor Necrosis Factor-Beta Receptor (TNF-β receptor); Tumor Necrosis Factor-Gamma Receptor (TNF-γ receptor); Transferrin Receptor; or the Vascular Endothelial Growth Factor Receptor (VEGF receptor).

9. The diabody complex of claim 8 , wherein said tumor-associated antigen is HER-2/neu.

10. The diabody complex of claim 8 , wherein said tumor-associated antigen is CD32B.

11. The diabody complex of claim 1 , wherein said third polypeptide chain comprises, in the N-terminal to C-terminal direction, said coil domain and said polypeptide portion of said protein that binds to a said serum albumin.

12. The diabody complex of claim 1 , wherein said third polypeptide chain comprises, in the N-terminal to C-terminal direction, said polypeptide portion of said protein that binds to said serum albumin and said coil domain.

Assignees

• Johnson Leslie S
• Huang Ling
• Rainey Godfrey Jonah Anderson
• Macrogenics Inc

Inventors

• Johnson Leslie S
• Huang Ling
• Rainey Godfrey Jonah Anderson

Classifications

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title

• C07K2317/31

  multispecific · CPC title

• C07K16/32

  against translation products of oncogenes · CPC title

• A61K39/39558 Primary

  against tumor tissues, cells, antigens · CPC title

• A61K38/164

  from bacteria · CPC title