BISPECIFIC ANTIBODY

• US 20150284475A1
• Filed: 11/21/2012
• Published: 10/08/2015
• Est. Priority Date: 11/21/2012
• Status: Active Grant

• Current Assignee: Wuhan YZY Biopharma Co., Ltd.
• Original Assignee: Wuhan YZY Biopharma Co., Ltd.
• Inventors: Zhou, Pengfei, Zhang, Jing, Yan, Yongxiang
• Granted Patent: US 9,562,110 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/35 Valency
• CPC Class Codes: C07K 2317/51 Complete heavy chain or Fd fragment, i.e. VH + CH1
• CPC Class Codes: C07K 2317/515 Complete light chain, i.e. VL + CL
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/64 comprising a combination of variable region and constant region components
• CPC Class Codes: C07K 2317/732 Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2317/94 Stability, e.g. half-life, pH, temperature or enzyme-resistance

Abstract

Provided are bispecific antibodies comprised of a single-chain unit having specificity to an immune cell and a monovalent unit having specificity to a tumor cell or a microorganism. The single-chain unit includes a single-chain variable fragment (scFv) fused to an Fc fragment and the monovalent unit includes a light chain and heavy chain pair. Also provided are methods of preparing bispecific antibodies and pharmaceutical and diagnostic uses of these antibodies.

Claims

1. An antibody comprising:

(a) a monovalent unit comprising a light chain-heavy chain pair having specificity to a tumor antigen selected from the group consisting of EGFR, Her2, EpCAM, CD20, CD30, CD33, CD47, CD52, CD133, CEA, gpA33, Mucins, TAG-72, CIX, PSMA, folate-binding protein, GD2, GD3, GM2, VEGF, VEGFR, Integrin, ERBB2, ERBB3, MET, IGF1R, EPHA3, TRAILR1, TRAILR2, RANKL, FAP and Tenascin; and

(b) a single-chain unit comprising a fusion peptide comprising a single-chain variable fragment (scFv) having specificity to an antigen selected from the group consisting of CD3, CD16, CD19, CD28 and CD64, wherein the scFv has Formula (I);

VH—

X—

VL-Fc, and wherein X is a linker.

4. The antibody of claim 1, wherein the light chain-heavy chain pair has specificity to a protein that is overexpressed on a tumor cell as compared to a corresponding non-tumor cell.

7. The antibody of claim 1, wherein the immune cell is selected from the group consisting of a T cell, a B cell, a monocyte, a macrophage, a neutrophil, a dendritic cell, a phagocyte, a natural killer cell, an eosinophil, a basophil, and a mast cell.

9. The antibody of claim 1, wherein the light chain is bound to the heavy chain through a disulfide bond.

10. The antibody of claim 1, wherein the heavy chain is bound to the fusion peptide through one or more disulfide bonds.

11. The antibody of claim 1, wherein the heavy chain comprises a human or a humanized Fc fragment.

12. The antibody of claim 11, wherein the Fc fragment of the heavy chain comprises a human IgG Fc fragment.

13. The antibody of claim 1, wherein the Fc fragment of the fusion peptide comprises a human or a humanized Fc fragment.

14. The antibody of claim 13, wherein the Fc fragment of the fusion peptide comprises a human IgG Fc fragment.

15. The antibody of claim 1, wherein the heavy chain and/or the Fc fragment of the fusion peptide comprise one or more substitutions, as compared to a wild-type antibody fragment, that form an ionic bond between the heavy chain and the Fc fragment.

16. The antibody of claim 15, wherein the substitutions are selected from Table 2.

17. The antibody of claim 1, wherein the heavy chain and/or the Fc fragment of the fusion peptide comprises one or more substitutions, as compared to a wild-type antibody fragment, that form a knob-into-the-hole conformational pairing between the heavy chain and the Fc fragment.

18. The antibody of claim 17, wherein the substitutions are selected from Table 3.

19. The antibody of claim 1, further comprising a detectable label.

20. The antibody of claim 1, wherein the CH2 domain is located between the scFv fragment and the CH3 domain.

21. The antibody of claim 1, wherein the fusion peptide does not contain a CH1 domain.

22. A composition comprising an antibody of claim 1 and a carrier.

23. The composition of claim 22, wherein the carrier is a pharmaceutical carrier.

24. A complex comprising an antibody of claim 1 bound to one or more antigens.

2-3. -3. (canceled)

5-6. -6. (canceled)

8. (canceled)

25-26. -26. (canceled)

27. An antibody comprising:

(a) a monovalent unit comprising a light chain-heavy chain pair having specificity to a tumor antigen selected from the group consisting of CD20, CD133 and EGFR; and

(b) a single-chain unit comprising a fusion peptide comprising a single-chain variable fragment (scFv) having specificity to CD3, wherein the scFv has Formula (I);

VH—

X—

VL-Fc, and wherein X is a linker.