Bispecific antigen binding protein complex and preparation methods of bispecific antibodies

Patent metadata

Field	Value
Publication number	US-9932412-B2
Application number	US-201314069091-A
Country	US
Kind code	B2
Filing date	Oct 31, 2013
Priority date	Oct 31, 2012
Publication date	Apr 3, 2018
Grant date	Apr 3, 2018

Abstract

A bispecific antigen binding protein complex comprising a first polypeptide comprising a first antigen binding site at an N terminus; a second polypeptide comprising a second antigen binding site at an N terminus; and a linker connecting the first polypeptide and the second polypeptide; wherein the linker comprises a tag at one terminus thereof, and wherein the tag is connected to a C-terminus of the first polypeptide or to an N-terminus of the second polypeptide, and comprises a cleavable amino acid sequence; as well as related compositions and methods.

First claim

What is claimed is:

1. A bispecific antigen binding protein complex comprising:
a first polypeptide comprising a first antigen binding site at an N terminus;
a second polypeptide comprising a second antigen binding site at an N terminus; and
a peptide linker connecting the first polypeptide and the second polypeptide; wherein the linker consists of 2 to 50 amino acids;
wherein the peptide linker comprises a tag attached to at least one terminus thereof, and wherein the tag is connected to at least one of a C-terminus of the first polypeptide and an N-terminus of the second polypeptide, and comprises a protease-cleavable amino acid sequence.

2. The protein complex of claim 1 , wherein the peptide linker includes a first tag at one terminus of the linker and a second tag at another terminus of the peptide linker, and wherein the first tag is connected to a C-terminus of the first polypeptide, the second tag is connected to an N-terminus of the second polypeptide, and the first tag and the second tag each includes a protease-cleavable amino acid sequence.

3. The protein complex of claim 1 , wherein at least one of the first polypeptide and the second polypeptide comprises an antibody heavy chain, an antibody light chain, a single-domain antibody, or an antibody fragment selected from a group consisting of Fab, Fab′, Fv, and scFv.

4. The protein complex of claim 1 , wherein the tag is selected from a group consisting of ubiquitin, ubiquitin-like protein, and TEV cleavage peptide.

5. The protein complex of claim 1 , wherein the first antigen binding site and the second antigen binding site each independently comprises a site binding specifically to a target antigen selected from the group consisting of VEGF, EGFR, EpCAM, CCRS, CD19, HER-2 neu, HER-3, HER-4, EGFR, PSMA, CEA, MUC-1 (mucin), MUC2, MUC3, MUC4, MUCS AC, MUC5 B, MUC7, βhCG, Lewis-Y, CD20, CD33, CD30, ganglioside GD3, 9-O-acetyl-GD3, GM2, Globo H, fucosyl GM1, poly SA, GD2, Carboanhydrase IX (MN/CA IX), CD44v6, Sonic Hedgehog (Shh), Wue-1, Plasma Cell Antigen, (membrane bound) IgE, Melanoma Chondroitin Sulfate Proteoglycan (MCSP), CCR8, TNF-alpha precursor, STEAP, mesothelin, A33 antigen, Prostate Stem Cell Antigen (PSCA) antigen, Ly-6, desmoglein 4, E-cadherin neoepitope, Fetal Acetylcholine Receptor, CD25, CA19-9 marker, CA-125 marker, Mullerian Inhibitory Substance (MIS) II receptor, sTn (sialyated Tn antigen; TAG-72), FAP (fibroblast activation antigen), endosialin, EGFRvIII, LG, SAS, and CD63.

6. The protein complex of claim 1 , wherein the first and second polypeptides each comprise an amino acid sequence independently selected from the group consisting of SEQ ID NOs: 8 to 44.

7. A polynucleotide encoding the protein complex of claim 1 .

8. The polynucleotide of claim 7 , wherein the polynucleotide comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 45 to 81.

9. A method of preparing a bispecific protein complex, the method comprising:
transforming a host cell with a recombinant expression vector comprising a polynucleotide of claim 7 ;
culturing the transformed host cell so as to express a bispecific protein complex; and
isolating the bispecific protein complex.

10. The method of claim 9 , wherein the tag is cleaved.

11. A method for treatment of a disease in a subject, comprising:
administering a bispecific protein complex of claim 1 to the subject, wherein the disease is a proliferative disorder, a neoplastic disease, an inflammatory disease, an autoimmune disease, an infectious disease, a viral disease, an allergic condition, a graft-versus-host disease, or a host-versus-graft disease.

12. The method of claim 11 , wherein the protein complex is bound to a second active agent, and targets the second active agent to a disease site.

13. A method for diagnosing a disease comprising obtaining a biological sample from a subject and contacting the biological sample with a composition comprising a bispecific protein complex of claim 1 , wherein the composition can detect an antigen specifically found in a disease, and wherein the disease is selected from the group consisting of a proliferative disorder, a neoplastic disease, an inflammatory disease, an autoimmune disease, an infectious disease, a viral disease, an allergic condition, a graft-versus-host disease, and a host-versus-graft disease.

14. The method of claim 13 , wherein the composition comprises a detectable label attached to the bispecific protein complex.

15. A method for diagnosing a disease in a subject comprising injecting the subject with a composition comprising a bispecific protein complex of claim 1 , wherein the composition can detect an antigen specifically found in a disease, and wherein the disease is selected from the group consisting of a proliferative disorder, a neoplastic disease, an inflammatory disease, an autoimmune disease, an infectious disease, a viral disease, an allergic condition, a graft-versus-host disease, and a host-versus-graft disease.

16. The method of claim 15 , wherein the composition comprises a detectable label attached to the bispecific protein complex.

17. A composition comprising the protein complex of claim 1 and a detectable label attached to the protein complex.

18. The protein complex of claim 1 , wherein the first polypeptide and second polypeptide each comprise a Fc region.

19. The protein complex of claim 18 , wherein the Fc region comprises a hinge region and CH2 and CH3 region.

Assignees

• Samsung Electronics Co Ltd

Inventors

• Kim Min-Kyung
• Lee Jungwook
• Hwang Sujeong
• Lee Jaeil

Classifications

• A61P37/06

  Immunosuppressants, e.g. drugs for graft rejection · CPC title

• A61P37/08

  Antiallergic agents (antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14) · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P37/02

  Immunomodulators · CPC title

• A61P37/00

  Drugs for immunological or allergic disorders · CPC title