US 12,227,594 B2
Bispecific 2+1 contorsbodies
Maria Amann, Schlieren (CH); Claudia Ferrara Koller, Schlieren (CH); Reto Flury, Schlieren (CH); Guy Georges, Penzberg (DE); Sandra Grau-Richards, Schlieren (CH); Alexander Haas, Penzberg (DE); Friederike Hesse, Penzberg (DE); Sabine Imhof-Jung, Penzberg (DE); and Christian Klein, Schlieren (CH)
Assigned to Hoffmann-La Roche Inc., Little Falls, NJ (US)
Appl. No. 16/760,820
Filed by Hoffmann-La Roche Inc., Little Falls, NJ (US)
PCT Filed Oct. 31, 2018, PCT No. PCT/EP2018/079785
§ 371(c)(1), (2) Date Apr. 30, 2020,
PCT Pub. No. WO2019/086500, PCT Pub. Date May 9, 2019.
Claims priority of application No. 17199537 (EP), filed on Nov. 1, 2017.
Prior Publication US 2021/0324108 A1, Oct. 21, 2021
Int. Cl. C07K 16/40 (2006.01); A61K 45/06 (2006.01); C07K 16/28 (2006.01); C07K 16/30 (2006.01)
CPC C07K 16/40 (2013.01) [A61K 45/06 (2013.01); C07K 16/2878 (2013.01); C07K 16/30 (2013.01); C07K 2317/31 (2013.01); C07K 2317/524 (2013.01); C07K 2317/526 (2013.01); C07K 2317/53 (2013.01); C07K 2317/55 (2013.01); C07K 2317/565 (2013.01); C07K 2319/30 (2013.01)] 14 Claims
 
1. A bispecific antibody comprising two fusion polypeptides and comprising 2+1 binding domains binding domain capable of specific binding to a tumor-specific antigen comprising:
(a)
(1) a first fusion polypeptide from N-terminus to C-terminus comprising a first heavy chain variable domain (first VH), a CH1 domain, a first peptide linker, a spacer domain, a second peptide linker, a first light chain variable domain (first VL), a CL domain (Ckappa), a third peptide linker, a second light chain variable domain (second VL), and a CH1 domain, wherein the first VH and the first VL forms a first binding domain, and
(2) a second fusion polypeptide from N terminus to C-terminus comprising a first heavy chain variable domain (first VH), a CH1 domain, a first peptide linker, a spacer domain, a second peptide linker, a first light chain variable domain (first VL), a CL domain (Ckappa), a third peptide linker, a second heavy chain variable domain (second VH), and a CL domain (Ckappa),
wherein the first VH and the first VL forms a first binding domain, and wherein the second VH and the second VL domain forms a second antigen binding domain, wherein the spacer domain comprises a hinge, a CH2 domain and a CH3 domain or a fragment thereof;
or
(b)
(1) a first fusion polypeptide from N-terminus to C-terminus comprising a first heavy chain variable domain (first VH), a CL domain (Ckappa), a first peptide linker, a spacer domain, a second peptide linker, a first light chain variable domain (first VL), a CH1 domain, a third peptide linker, a second heavy chain variable domain (second VH), and a CH1 domain, wherein the first VH and the first VL forms a first binding domain, and
(2) a second fusion polypeptide from N-terminus to C-terminus comprising a first heavy chain variable domain (first VH), a CL domain (Ckappa), a first peptide linker, a spacer domain, a second peptide linker, a first light chain variable domain (first VL), a CH1 domain, a third peptide linker, a second light chain variable domain (second VL), and a CL domain (Ckappa),
wherein the first VH and the first VL forms a first binding domain, and wherein the second VH and the second VL domain forms a second antigen binding domain, wherein the spacer domain comprises a hinge, a CH2 domain and a CH3 domain or a fragment thereof,
or
(c)
(1) a first fusion polypeptide from N-terminus to C-terminus comprising a first light chain variable domain (first VL), a CL domain (Ckappa), a first peptide linker, a spacer domain, a second peptide linker, a first heavy chain variable domain (first VH), a CH1 domain, a third peptide linker, a second light chain variable domain (second VL), and a CH1 domain, wherein the first VH and the first VL forms a first binding domain, and
(2) a second fusion polypeptide from N-terminus to C-terminus comprising a first light chain variable domain (first VL), a CL domain (Ckappa), a first peptide linker, a spacer domain, a second peptide linker, a first heavy chain variable domain (first VH), a CH1 domain, a third peptide linker, a second heavy chain variable domain (second VH), and a CL domain (Ckappa),
wherein the first VH and the first VL forms a first binding domain, and wherein the second VH and the second VL domain forms a second antigen binding domain, wherein the spacer domain comprises a hinge, a CH2 domain and a CH3 domain or a fragment thereof,
or
(d)
(1) a first fusion polypeptide from N-terminus to C-terminus comprising a second light chain variable domain (second VL), a CH1 domain, a first peptide linker, a first heavy chain variable domain (first VH), a CL domain (Ckappa) a second peptide linker, a spacer domain, a third peptide linker, a first light chain variable domain (first VL) and a CH1 domain, wherein the first VH and the first VL forms a first binding domain, and
(2) a second fusion polypeptide from N-terminus to C-terminus comprising a second heavy chain variable domain (second VH), a CL domain (Ckappa), a first peptide linker, a first heavy chain variable domain (first VH), a CL domain (Ckappa), a second peptide linker, a spacer domain, a third peptide linker, a first light chain variable domain (first VL) and a CH1 domain,
wherein the first VH and the first VL forms a first binding domain, and wherein the second VH and the second VL domain forms a second antigen binding domain, wherein the spacer domain comprises a hinge, a CH2 domain and a CH3 domain or a fragment thereof;
wherein for each of (a)-(d) above, the first and second antigen binding domain are associated with each other to form a circular fusion polypeptide, and wherein the spacer domain of the first fusion polypeptide and the spacer domain of the second fusion polypeptide are associated covalently to each other by a disulfide bond and comprise at least one modification promoting the association of the first fusion polypeptide and second fusion polypeptide.