Binding domain-immunoglobulin fusion proteins
View Patent ↗The invention relates to novel binding domain-immunoglobulin fusion proteins that feature a binding domain for a cognate structure such as an antigen, a counterreceptor or the like, a hinge region polypeptide having either zero or one cysteine residue, and immunoglobulin CH2 and CH3 domains, and that are capable of ADCC and/or CDC while occurring predominantly as monomeric polypeptides. The fusion proteins can be recombinantly produced at high expression levels. Also provided are related compositions and methods, including immunotherapeutic applications.
1. A method of treating a malignancy or immune system disorder, comprising administering to a patient in need thereof a therapeutically effective amount of a single chain protein that binds to a target biological molecule, wherein the single chain protein comprises from amino terminus to carboxy terminus:
(a) an immunoglobulin binding domain polypeptide that specifically binds to the target biological molecule,
(b) an IgG1 hinge peptide in which the number of cysteines is one,
(c) an immunoglobulin heavy chain CH2 constant region polypeptide, and
(d) an immunoglobulin heavy chain CH3 constant region polypeptide, and wherein the target biological molecule is associated with a malignancy or immune system disorder.
2. The method of claim 1 , wherein the immunoglobulin binding domain polypeptide is a single chain Fv polypeptide.
3. The method of claim 2 , wherein the heavy and light chain variable regions of the single chain Fv are joined by a polypeptide linker of about 5 amino acids to about 15 amino acids.
4. The method of claim 3 , wherein the linker comprises an amino acid sequence of Gly-Gly-Gly-Gly-Ser (SEQ ID NO:39).
5. The method of claim 4 , wherein the immunoglobulin heavy chain CH2 and CH3 constant region polypeptides are human IgG1 CH2 and CH3 constant region polypeptides.
6. The method of claim 1 , wherein the immunoglobulin heavy chain CH2 and CH3 constant region polypeptides are human CH2 and CH3 constant region polypeptides.
7. The method of claim 6 , wherein the human CH2 and CH3 constant region polypeptides are IgG1 CH2 and CH3 constant region polypeptides.
8. The method of claim 1 , wherein the target biological molecule comprises a B-cell molecule.
9. The method of claim 8 , wherein the single chain protein is capable of depleting a population of target B-cells.
10. The method of claim 8 , wherein the B-cell molecule is CD20.
11. The method of claim 1 , wherein the binding domain polypeptide is a single chain Fv capable of binding human CD20, and the immunoglobulin heavy chain CH2 and CH3 constant region polypeptides are human IgG1 CH2 and CH3 constant region polypeptides.
12. The method of claim 11 , wherein the single chain Fv is derived from the heavy chain and light chain variable regions of monoclonal antibody 2H7.
13. The method of claim 11 , wherein the heavy chain CH2 constant region polypeptide comprises a CH2 domain in which a leucine has been replaced with serine at position 234.
14. The method of claim 8 , wherein the B-cell molecule is CD37.
15. The method of claim 1 , wherein the binding domain polypeptide is a single chain Fv capable of binding a human CD37, and the immunoglobulin heavy chain CH2 and CH3 constant region polypeptides are IgG1 CH2 and CH3 constant region polypeptides.
16. The method of claim 15 , wherein the single chain protein Fv is derived from the heavy chain and light chain variable regions of monoclonal antibody G28-1.
17. The method of claim 15 , wherein the heavy chain CH2 constant region polypeptide comprises a CH2 domain in which a leucine has been replaced with serine at position 234.
18. The method of claim 8 , wherein the B-cell molecule is selected from the group consisting of CD19, CD22, CD30 ligand, and CD54.
19. The method of claim 1 , wherein the target biological molecule is selected from the group consisting of CD2, CD5, CD10, CD27, CD28, CD40, CD40 ligand, CD43, CD59, CD48, CD72, CD70, CD83, CD86/B7.2, CD106, CTLA-4, DEC-205, 4-1BB, 4-1BB ligand, interferon-γ, interleukin-4, interleukin-12, interleukin-17, and VLA-4 (α 4 β 7 ).
20. The method of claim 1 , wherein the target biological molecule is selected from the group consisting of HER1, HER2, HER3, HER4, vascular endothelial cell growth factor, insulin-like growth factor-I, insulin-like growth factor-II, MUC-1, NY-ESO-1, NA 17-A, Melan-A/MART-1, tyrosinase, Gp-100, MAGE, BAGE, GAGE, the HOM-MEL-40 antigen encoded by the SSX2 gene, carcinoembyonic antigen, and PyLT.
21. The method of claim 1 , wherein the target biological molecule is selected from the group consisting of an interleukin-17 receptor, an epidermal growth factor receptor, a vascular endothelial cell growth factor receptor, a transferrin receptor, an estrogen receptor, a progesterone receptor, a follicle stimulating hormone receptor, a retinoic acid receptor, and any of the CTA class receptors.
22. The method according to any one of claims 1 - 21 , wherein the immunoglobulin binding domain polypeptide is humanized.
23. The method according to any one of claims 1 , 2 , 6 or 7 , wherein the single chain protein promotes antibody dependent cell-mediated cytotoxicity, complement fixation, or both.
24. The method of claim 1 , wherein the malignancy is a B-cell malignancy.
25. The method of claim 24 , wherein the B-cell malignancy is a B-cell lymphoma or chronic lymphocytic leukemia.
26. The method of claim 1 , wherein the immune system disorder is a B-cell disorder.
27. The method of claim 26 , wherein the B-cell disorder is psoriasis, systemic lupus erythematosus, type I diabetes mellitus, multiple sclerosis, immune thrombocytopenic purpura, inflammatory bowel disease, Crohn's disease, or ulcerative colitis.
28. The method of claim 26 , wherein the B-cell disorder is rheumatoid arthritis.
29. The method of claim 1 , wherein the single chain protein binds to a cancer cell molecule.