Multispecific domain exchanged common variable light chain antibodies

• US 10,323,099 B2
• Filed: 04/07/2016
• Issued: 06/18/2019
• 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, Neumann, Christiane, Klein, Christian, Moessner, Ekkehard, Schaefer, Wolfgang
• Primary Examiner(s): Bristol, Lynn A
• Application Number:
  US15/092,847
• Publication Number:
  US 20160319036A1
• Time in Patent Office: 1,167 Days
• Field of Search: 4241331
• 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

Multispecific antibodies based on antibodies that share a common light chain are described herein. The multispecific antibodies include: modified heavy chains having, by domain exchange, a common light chain variable domain VL; and two modified light chains having, 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 such antibodies and their use.

Claims

1. A multispecific antibody, comprising:

(a) a first modified light chain, wherein the first modified light chain comprises in C-terminal to N-terminal direction;

a constant light chain domain of kappa isotype (CLκ

); and

a variable heavy chain domain (VH₁) derived from an antibody which specifically binds to a first antigen;

(b) a second modified light chain, wherein the second modified light chain comprises in C-terminal to N-terminal direction;

a constant light chain domain of lambda isotype (CLλ

); and

a variable heavy chain domain (VH₂) derived from an antibody which specifically binds to a second antigen; and

(c) two modified heavy chains, wherein each modified heavy chain comprises in C-terminal to N-terminal direction;

heavy chain constant domains 3 and 2 (CH3 and CH2);

a hinge region;

a heavy chain constant domain 1 (CH1); and

a light chain variable domain (VL),

wherein the VL in one of the two modified heavy chains and the VL in the other of the two modified heavy chains are identical,

wherein the first modified light chain of (a) pairs with one of the two modified heavy chains to form a first antigen binding site which specifically binds to the first antigen, and

wherein the second modified light chain of (b) pairs with the other of the two modified heavy chains to form a second antigen binding site which specifically binds to the 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 pharmaceutical composition comprising the multispecific antibody according to claim 1 and at least one pharmaceutically acceptable excipient.

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

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

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

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

10. An expression vector containing a nucleic acid that encodes the multispecific antibody of claim 1.

11. A prokaryotic or eukaryotic host cell comprising an expression vector according to claim 10.

12. 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.