	US 12,435,137 B2
	Antigen-binding molecule containing two antigen-binding domains that are linked to each other
Hirotake Shiraiwa, Shizuoka (JP); and Tatsuya Kawa, Shizuoka (JP)
Assigned to CHUGAI SEIYAKU KABUSHIKI KAISHA, Tokyo (JP)
Appl. No. 17/264,388
Filed by Chugai Seiyaku Kabushiki Kaisha, Tokyo (JP)
PCT Filed Aug. 2, 2019, PCT No. PCT/JP2019/030564 § 371(c)(1), (2) Date Jan. 29, 2021, PCT Pub. No. WO2020/027330, PCT Pub. Date Feb. 6, 2020.
Claims priority of application No. 2018-146929 (JP), filed on Aug. 3, 2018.
Prior Publication US 2022/0195045 A1, Jun. 23, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C07K 16/28 (2006.01); C12N 15/63 (2006.01)

CPC C07K 16/2809 (2013.01) [C07K 16/2818 (2013.01); C12N 15/63 (2013.01); C07K 2317/31 (2013.01); C07K 2317/52 (2013.01); C07K 2317/55 (2013.01); C07K 2317/569 (2013.01); C07K 2317/75 (2013.01); C07K 2317/76 (2013.01)]

12 Claims

1. An antigen-binding molecule comprising a first antigen-binding domain crosslinked with a second antigen-binding domain,

wherein the first antigen-binding domain and the second antigen-binding domain domains comprise a hinge region,

wherein the first and second antigen-binding domains each comprise an antibody fragment which binds to a particular antigen,

wherein the first and second antigen-binding domains are linked with each other by two or more bonds between amino acid residues in the first and second antigen binding domains,

wherein at least one of the bonds is a disulfide bond, and

wherein at least one amino acid residue from which the two or more bonds originate is present within a CH1 region and/or within a CL region of the antibody fragment of the first and second antigen-binding domains and at least one amino acid residue from which the bonds originate is present within a hinge region, wherein at least one of the bonds linking the two antigen-binding domains is formed by:

linking an amino acid residue in a CH1 region of the first antigen-binding domain with an amino acid residue in a CH1 region of the second antigen-binding domain,

wherein at least one of the bonds linking the two antigen-binding domains is formed by linking any two amino acid residues selected from the group consisting of positions 119 to 123, 131 to 140, 148 to 150, 155 to 167, 174 to 178, 188 to 197, and 201 to 214, according to EU numbering, in a CH1 region of the first antigen-binding domain and a CH1 region of the second antigen binding domain,

linking an amino acid residue in a CL region of the first antigen-binding domain with an amino acid residue in a CL region of the second antigen-binding domain,

wherein at least one of the bonds linking the two antigen-binding domains is formed by linking any two amino acid residues selected from the group consisting of positions 108, 109, 112, 121, 123, 126, 128, 151, 152, 153, 156, 184, 186, 188, 189, 190, 195, 196, 200, 201, 202, 203, 208, 210, 211, 212, and 213, according to Kabat numbering, in a CL region of the first antigen-binding domain and a CL region of the second antigen binding domain, or

linking an amino acid residue in a CH1 region of the first antigen-binding domain with an amino acid residue in a CL region of the second antigen-binding domain,

wherein the amino acid residue in a CH1 region is selected from the group consisting of positions 188, 189, 190, 191, 192, 193, 194, 195, 196, and 197, according to EU numbering, and the amino acid residue in a CL region is selected from the group consisting of positions 121, 122, 123, 124, 125, 126, 127, and 128, according to Kabat numbering, and

wherein said antigen-binding molecule has increased resistance to protease cleavage as compared to a control antigen-binding molecule, wherein the control antigen-binding molecule differs from the antigen-binding molecule only in that the control antigen-binding molecule has one less bond between the two antigen-binding domains.