Fc-free antibodies comprising two Fab-fragments and methods of use

• US 20160130347A1
• Filed: 10/03/2013
• Published: 05/12/2016
• Est. Priority Date: 10/08/2012
• Status: Active Grant

• Current Assignee: Roche Glycart AG (Roche Holding AG)
• Original Assignee: Roche Glycart AG (Roche Holding AG)
• Inventors: Bruenker, Peter, Fauti, Tanja, Jaeger, Christiane, Klein, Christian, Umaa, Pablo
• Granted Patent: US 10,087,250 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/39541 against normal tissues, cells
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 16/3053 Skin, nerves, brain
• CPC Class Codes: C07K 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/54 F(ab')2
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/66 comprising a swap of domains, e.g. CH3-CH2, VH-CL or VL-CH1
• CPC Class Codes: C07K 2317/75 Agonist effect on antigen
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2317/94 Stability, e.g. half-life, pH, temperature or enzyme-resistance
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

The present invention relates to bispecific antibodies comprising at least two Fab fragments, wherein the first Fab fragment comprises at least one antigen binding site specific for a first antigen; and the second Fab fragment comprises at least one antigen binding site specific for a second antigen, wherein either the variable regions or the constant regions of the second Fab heavy and light chain are exchanged and the two Fab fragments are connected to each other by two peptide linkers; and wherein the bispecific antibody is devoid of a Fc domain; methods for their production, pharmaceutical compositions containing said antibodies, and uses thereof.

Claims

1. A bispecific antibody comprising at least two Fab fragments, wherein

the first Fab fragment comprises at least one antigen binding site specific for a first antigen;

and the second Fab fragment comprises at least one antigen binding site specific for a second antigen wherein either the variable regions or the constant regions of the second Fab heavy and light chain are exchanged; and

the two Fab fragments are connected to each other by two peptide linkers and wherein the bispecific antibody is devoid of a Fc domain.

2. The bispecific antibody of claim 1, additionally comprising a third Fab fragment.

3. The bispecific antibody of claim 2, wherein the third Fab fragment comprises at least one antigen binding site specific for the first or second antigen.

4. The bispecific antibody of claim 3, wherein the third Fab fragment comprises at least one antigen binding site specific for the first antigen.

5. The bispecific antibody of any of claims 2 to 4, wherein the third Fab fragment is connected to the first Fab fragment by one or two peptide linkers.

6. The bispecific antibody of claims 2 to 4, wherein the third Fab fragment is connected to the second Fab fragment by one or two peptide linkers.

7. The bispecific antibody of any of claims 1 to 6, wherein the first Fab fragment comprises at least one antigen binding site specific for a Tumor Antigen;

and the second Fab fragment comprises at least one antigen binding site specific for a T-cell activating antigen.

8. The bispecific antibody of claim 7, wherein the T-cell activating antigen is a CD3 T-Cell Co-Receptor (CD3) antigen.

9. The bispecific antibody of any of claim 7 or 8, wherein the Tumor Antigen is selected from the group of Melanoma-associated Chondroitin Sulfate Proteoglycan (MCSP), Epidermal Growth Factor Receptor (EGFR), Carcinoembryonic Antigen (CEA), Fibroblast Activation Protein (FAP) and CD33.

10. The bispecific antibody of claim 9, wherein the Tumor Antigen is MCSP.

11. A pharmaceutical composition comprising a bispecific antibody of claims 1 to 10.

12. The bispecific antibody of claims 1 to 10 for the treatment of cancer.

13. The bispecific antibody of claims 1 to 10 for use as a medicament.

14. Use of the bispecific antibody of claims 1 to 10 in the manufacture of a medicament.

15. The use of claim 14, wherein the medicament is for treatment of cancer.

16. A prokaryotic or eukaryotic host cell comprising vectors comprising nucleic acid molecules encoding the light chains and heavy chains of the bispecific antibody of any of claims 1 to 10.

17. A method of producing an antibody comprising culturing the host cell of claim 16 so that the antibody is produced.

18. The invention as described herein, especially with reference to the foregoing examples.