MULTISPECIFIC DOMAIN EXCHANGED COMMON VARIABLE LIGHT CHAIN ANTIBODIES

• US 20160319036A1
• Filed: 04/07/2016
• Published: 11/03/2016
• Est. Priority Date: 10/11/2013
• Status: Active Grant

• Current Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: BRUENKER, PETER, JAEGER, CHRISTIANE, KLEIN, CHRISTIAN, MOESSNER, EKKEHARD, SCHAEFER, WOLFGANG
• Granted Patent: US 10,323,099 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/2878 against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
• CPC Class Codes: C07K 16/3053 Skin, nerves, brain
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/35 Valency
• CPC Class Codes: C07K 2317/41 Glycosylation, sialylation, or fucosylation
• 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 2317/522 CH1 domain
• CPC Class Codes: C07K 2317/524 CH2 domain
• CPC Class Codes: C07K 2317/526 CH3 domain
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/60 characterized by non-natural combinations of immunoglobulin fragments
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation

Abstract

The present invention relates to multispecific antibodies based on antibodies that share a common light chain. The multispecific antibodies include modified heavy chains comprising by domain exchange a common light chain variable domain VL; and two modified light chains comprising by domain exchange variable heavy chain domains of a first antibody (VH₁) and a second antibody (VH₂), wherein one light chain is of kappa isotype and one light chain is of lambda isotype. The present invention also relates to methods for the manufacture of said antibodies and their use.

Claims

1. A multispecific antibody, comprising:

a) two modified heavy chains;

wherein each heavy chain comprises in C-terminal to N-terminal direction heavy chain constant domains 3, 2 and 1 (CH3, CH2 and CH1) and a light chain variable domain (VL), wherein the light chain variable domain (VL) is the variable domain of a common light chain;

b) one modified light chain, wherein the modified light chain comprises in C-terminal to N-terminal direction a constant light chain domain (CL) of kappa isotype and a variable heavy chain domain (VH₁) derived from an antibody, which specifically binds to a first antigen; and

c) one modified light chain, wherein the modified light chain comprises in C-terminal to N-terminal direction a constant light chain domain of lambda isotype and a variable heavy chain domain (VH₂) derived from an antibody, which specifically binds to a second antigen;

wherein the variable domains VH₁ and VL form a first antigen binding site which specifically binds to a first antigen, and wherein the variable domains VH₂ and VL form a second antigen binding site which specifically binds to second antigen.

2. The multispecific antibody according to claim 1, wherein the antibody is a bivalent, bispecific antibody.

3. The multispecific antibody according to claim 1, wherein the antibody is of IgG class.

4. The multispecific antibody according to claim 1, wherein the antibody is of IgG1 or IgG4 subclass.

5. A nucleic acid encoding a multispecific antibody according to claim 1.

6. An expression vector containing a nucleic acid according to claim 5 capable of expressing said nucleic acid in a prokaryotic or eukaryotic host cell.

7. A prokaryotic or eukaryotic host cell comprising an expression vector according to claim 6.

8. A method for the preparation of a multispecific antibody according to claim 1, comprising the steps of

a) transforming a host cell with expression vectors comprising nucleic acid molecules encoding a multispecific antibody according to claim 1;

b) culturing the host cell under conditions that allow synthesis of said multispecific antibody molecule; and

c) recovering said multispecific antibody molecule from said culture.

9. A pharmaceutical composition comprising the multispecific antibody according to claim 1 and at least one pharmaceutically acceptable excipient.

12. A method for the treatment of a patient in need of therapy, characterized by administering to the patient a therapeutically effective amount of a multispecific antibody according to claim 1.

17. A pharmaceutical composition comprising the multispecific antibody according to claim 2 and at least one pharmaceutically acceptable excipient.

18. A pharmaceutical composition comprising the multispecific antibody according to claim 3 and at least one pharmaceutically acceptable excipient.

19. A pharmaceutical composition comprising the multispecific antibody according to claim 4 and at least one pharmaceutically acceptable excipient.

20. A method for the treatment of a patient in need of therapy, characterized by administering to the patient a therapeutically effective amount of a multispecific antibody according to claim 2.

21. A method for the treatment of a patient in need of therapy, characterized by administering to the patient a therapeutically effective amount of a multispecific antibody according to claim 3.

22. A method for the treatment of a patient in need of therapy, characterized by administering to the patient a therapeutically effective amount of a multispecific antibody according to claim 4.

10-11. -11. (canceled)

13. A method for the generation of a multispecific antibody based on a first antibody, which specifically binds to a first antigen, and a second antibody, which specifically binds to a second antigen, wherein the first antibody and the second antibody comprise a common light chain, comprising the steps of

a) modifying the light chain derived from said first antibody to obtain a light chain comprising in C-terminal to N-terminal direction a constant light chain domain of kappa isotype and a heavy chain variable domain (VH₁) by replacing the light chain variable domain (VL) by the heavy chain variable domain (VH₁) of the heavy chain derived from said first antibody and, optionally, replacing the original constant light chain domain by a constant light chain domain of kappa isotype;

b) modifying the light chain derived from said second antibody to obtain a light chain comprising in C-terminal to N-terminal direction a constant light chain domain of lambda isotype and a heavy chain variable domain (VH₂) by replacing the light chain variable domain (VL) by the heavy chain variable domain (VH₂) of the heavy chain derived from said second antibody and, optionally, replacing the original constant light chain domain by a constant light chain domain of lambda isotype;

c) modifying the heavy chain derived from said first antibody in order to obtain a heavy chain comprising in C-terminal to N-terminal direction heavy chain constant domains 3, 2 and 1 (CH3, CH2 and CH1) and a light chain variable domain (VL) by replacing the heavy chain variable domain (VH₁) by the light chain variable domain (VL) of the light chain derived from said first antibody, and/or

modifying the heavy chain derived from said second antibody in order to obtain a heavy chain comprising in C-terminal to N-terminal direction heavy chain constant domains 3, 2 and 1 (CH3, CH2 and CH1) and a light chain variable domain (VL) by replacing the heavy chain variable domain (VH₂) by the light chain variable domain (VL) of the light chain derived from said second antibody.

14. The method of claim 13 further comprising the steps of

a) transforming a host cell with expression vectors comprising nucleic acid molecules encoding the obtained multispecific antibody,

b) culturing the host cell under conditions that allow synthesis of said multispecific antibody molecule;

and

c) recovering said multispecific antibody molecule from said culture.

15. A multispecific antibody obtained by the method of claim 13.

16. A multispecific antibody obtained by the method of claim 14.