MONOVALENT ASYMMETRIC BISPECIFIC ANTIBODIES WITH TANDEM FAB Russian patent published in 2022 - IPC A61K39/395 A61K51/10 C07K16/24 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology and is a monovalent asymmetric bispecific antibody with tandem Fab (MAT-Fab antibody), containing polypeptide chains (a), (b), (c), and (d), where (a) is a heavy polypeptide chain (heavy chain), wherein each heavy chain contains (in the direction from N-end to C-end): VL_A-CL-VH_B-CH1-hinge-CH2-CH3m1, where VL_A is a variable domain of a human immunoglobulin light chain, directly attached to CL, which is a constant domain of the human light chain, wherein VL_A-CL is a component of the immunoglobulin light chain of the first binding unit Fab recognizing the first antigen or epitope, and is directly attached to VH_B, wherein VH_B is a variable domain of the human immunoglobulin heavy chain, directly attached to CH1, which is a constant CH1 domain of the human immunoglobulin heavy chain, wherein VH_B-CH1 is a component of the immunoglobulin heavy chain of the second binding unit Fab recognizing the second antigen or epitope, wherein VH_B-CH1 is directly attached to the hinge-CH2 structure, wherein the hinge-CH2 structure is a hinge-CH2 region of the immunoglobulin heavy chain, and the hinge-CH2 structure is directly attached to CH3m1, which is the first constant CH3 domain of the human immunoglobulin heavy chain, mutated by one or more mutations providing the formation of “knob-into-hole” (KiH) structures with the formation of a structural knob or a structural hole in the specified constant domain CH3m1; (b) is the first light chain of MAT-Fab, containing VH_A-CH1, wherein VH_A is a variable domain of the human immunoglobulin heavy chain, directly attached to CH1, which is a constant CH1 domain of the human immunoglobulin heavy chain, and VH_A-CH1 is a component of the heavy chain of the specified first binding unit Fab; (c) is the second light chain of MAT-Fab, containing VL_B-CL, wherein VL_B is a variable domain of a human immunoglobulin light chain, directly attached to CL, which is CL domain of the human immunoglobulin light chain, and VL_B-CL is a component of the immunoglobulin light chain of the specified second binding unit Fab; and (d) is a polypeptide Fc chain containing hinge-CH2-CH3m2, where the hinge-CH2 structure is the hinge-CH2 region of the immunoglobulin heavy chain, and the hinge-CH2 structure is directly attached to CH3m2, which is the second constant CH3 domain of the human immunoglobulin heavy chain, mutated by one or more mutations providing the formation of “knob-into-hole” (KiH) structures with the formation of a structural knob or a structural hole in the specified constant domain CH3m2; provided that, when CH3m1 domain of the heavy chain is mutated to form a structural knob, CH3m2 Fc chain domain is mutated to form a complementary structural hole, which promotes the pairing of CH3m1 domain with CH3m2 domain; and, when CH3m1 domain of the specified heavy chain is mutated to form a structural hole, CH3m2 Fc chain domain is mutated to form a complementary structural knob, which promotes the pairing of CH3m1 domain with CH3m2 domain; and the specified MAT-Fab antibody optionally contains a mutation in CH3m1 domain and in CH3m2 domain, leading to the introduction of a cysteine residue promoting the formation of a disulfide bond, when CH3m1 domain is paired with CH3m2 domain.

EFFECT: invention makes it possible to increase the efficiency of tumor treatment.

71 cl, 6 dwg, 11 tbl, 1 ex