Activatable bispecific antibodies

Patent metadata

Field	Value
Publication number	US-2022017640-A1
Application number	US-202017091883-A
Country	US
Kind code	A1
Filing date	Nov 6, 2020
Priority date	Aug 24, 2010
Publication date	Jan 20, 2022
Grant date	—

Abstract

The current invention relates to bispecific antibodies wherein the binding affinity to one of the two antigens is reduced and which can be activated by tumor- or inflammation-specific proteases, and the preparation and use of such bispecific antibodies.

First claim

1 . A bispecific antibody comprising
a) a first antibody that binds to a first antigen comprising a VH 1 domain and a VL 1 domain, and b) a second antibody that binds to a second antigen,
wherein the VH 1 domain is fused N-terminally via a first peptide linker to the
second antibody, and the VL 1 domain is fused N-terminally via a second peptide linker to the second antibody, and

characterized in that
one of the linkers comprises a tumor- or inflammation-specific protease cleavage site, and the other linker does not comprise a protease cleavage site; and
the binding affinity of the bispecific antibody to the first antigen is reduced 5 times or more compared to the corresponding bispecific antibody in which the protease cleavage site is cleaved.

2 . The bispecific antibody according to claims 1 characterized in that the second antibody is a whole antibody; and
the VH 1 domain is fused N-terminally via the first linker to the C-terminus of the first heavy chain of the second antibody, and
the VL 1 domain is fused N-terminally via the second linker to the C-terminus of the second heavy chain of the second antibody.

3 . The bispecific antibody according to claim 2 characterized in comprising from
C- to N-terminus the following polypeptide chains
one VH 1 -peptide linker-CH3-CH2-CH1-VH 2 chain
two CL-VL 2 chains
one VL 1 -peptide linker-CH3-CH2-CH1-VH 2 chain

4 . The bispecific antibody according to claim 2 characterized in that
the first CH3 domain of the heavy chain of the whole antibody and the second CH3 domain of the whole antibody each meet at an interface which comprises an alteration in the original interface between the antibody CH3 domains;
wherein i) in the CH3 domain of one heavy chain,
an amino acid residue is replaced with an amino acid residue having a larger side chain volume, thereby generating a protuberance within the interface of the CH3 domain of one heavy chain which is positionable in a cavity within the interface of the CH3 domain of the other heavy chain and
ii) in the CH3 domain of the other heavy chain,
an amino acid residue is replaced with an amino acid residue having a smaller side chain volume, thereby generating a cavity within the interface of the second CH3 domain within which a protuberance within the interface of the first CH3 domain is positionable.

5 . The bispecific antibody according to claim 4 characterized in that
said amino acid residue having a larger side chain volume is selected from the group consisting of arginine (R), phenylalanine (F), tyrosine (Y), and tryptophan (W) and said amino acid residue having a smaller side chain volume is selected from the group consisting of alanine (A), serine (S), threonine (T), and valine (V).

6 . The bispecific antibody according to claim 5 characterized in that
both CH3 domains are further altered by the introduction of a cysteine (C) residue in positions of each CH3 domain such that a disulfide bridge between the CH3 domains can be formed.

7 . The bispecific antibody according to claim 1 , characterized in that
the VH 1 domain and the VL 1 domain are stabilized a) by a disulfide bridge; and/or b) by a CH1 domain and a CL domain

8 . The bispecific antibody according to claim 1 characterized in that
the second antibody is a Fv fragment; and
the VH 1 domain is fused N-terminally via the first linker to the C-terminus of the first chain of the second antibody Fv fragment, and
the VL 1 domain is fused N-terminally via a second linker to the C-terminus of the second chain of the second antibody Fv fragment.

9 . The bispecific antibody according to claim 8 characterized in that the first antibody is a whole antibody.

10 . The bispecific antibody according to claim 9 characterized in comprising from C- to N-terminus the following polypeptide chains
a) two CH3-CH2-CH1-VH 1 -peptide linker-VH 2 chains
two CL-VL 1 -peptide linker-VL 2 -chains; or

b) two CH3-CH2-CH1-VH 1 -peptide linker-VL 2 chains
two CL-VL 1 -peptide linker-VH 2 chains

11 . The bispecific antibody according to claim 8 , characterized in that
the VH 2 domain and the VL 2 domain are stabilized by a disulfide bridge.

12 . The bispecific antibody according to claim 1 characterized in that
the second antibody is a Fab fragment; and
the VH 1 domain is fused N-terminally via the first linker to the C-terminus of the first chain of the second antibody Fab fragment, and
the VL 1 domain is fused N-terminally via a second linker to the C-terminus of the second chain of the second antibody Fab fragment

13 . The bispecific antibody according to claim 12 characterized in that the first antibody is a whole antibody.

14 . The bispecific antibody according to claim 12 characterized in comprising from C-to N-terminus the following polypeptide chains
a) two CH3-CH2-CH1-VH 1 -peptide linker-CH1-VH 2 chains
two CL-VL 1 -peptide linker-CL-VL 2 -chains; or

b) two CH3-CH2-CH1-VH 1 -peptide linker-CL-VL 2 chains
two CL-VL 1 -peptide linker-CH1-VH 2 chains

15 . The bispecific antibody according to claim 12 characterized in that the first antibody is a Fv fragment.

16 . The bispecific antibody according to claim 15 characterized in comprising from C-to N-terminus the following polypeptide chains
a) one VH 1 -peptide linker-CH1-VH 2 chain
one VL 1 -peptide linker-CL-VL 2 chains; or

b) one VH 1 -peptide linker-CL-VL 2 chain
one VL 1 -peptide linker-CH1-VH 2 chains

17 . The bispecific antibody according to claim 1 characterized in that
the binding affinity of the bispecific antibody to the first antigen is reduced 10 times or more compared to the corresponding bispecific antibody in which the protease cleavage site is cleaved.

18 . A pharmaceutical composition comprising the antibody according to claim 1 .

19 - 22 . (canceled)

23 . Method of treatment of patient suffering from cancer by administering an antibody according to claim 1 to a patient in need of such treatment.

24 . (canceled)

Assignees

• Hoffmann La Roche

Inventors

• Brinkmann Ulrich
• Croasdale Rebecca
• Metz Silke
• Schanzer Juergen Michael
• Sustmann Claudio
• Umana Pablo

Classifications

• C07K16/2863 Primary

  against receptors for growth factors, growth regulators · CPC title

• C07K16/468 Primary

  Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies · CPC title

• C07K2319/00

  Fusion polypeptide · CPC title

• C07K2317/64

  comprising a combination of variable region and constant region components · CPC title

• C07K16/461

  Igs containing Ig-regions, -domains or -residues form different species · CPC title