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US 11,634,502 B2Granted
Heterodimeric bispecific antibodies
AssigneeAmgen Inc.
InventorsWei Yan, Martin J. Pentony, Luis G. Borges, Mark L. Michaels
Life of this patent
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Abstract
Provided herein are heterodimeric bispecific antibodies that can mediate cytolysis of a target cell by an immune effector cell, nucleic acids encoding such antibodies, methods of making such antibodies, and methods of using such antibodies. These antibodies comprise two different polypeptide chains, each comprising two immunoglobulin variable regions and, optionally, a half life-extending moiety.
Claims24
A heterodimeric bispecific antibody comprising
- (a) a first polypeptide chain comprising an amino acid sequence having the formula V1-L1-V2-L2-CH1, wherein V1 and V2 are immunoglobulin variable regions, L1 and L2 are linkers, L2 can be present or absent, and CH1 is a first immunoglobulin heavy chain constant region comprising SEQ ID NO:
- 70;
- and
- (b) a second polypeptide chain comprising an amino acid sequence having the formula V3-L3-V4-L4-CL, wherein V3 and V4 are immunoglobulin variable regions, L3 and L4 are linkers, L4 can be present or absent, and CL is an immunoglobulin light chain constant region comprising SEQ ID NO:
- 71 or 73;
- wherein either or both of the first polypeptide chain and the second polypeptide chain further comprise(s) an in vivo half life-extending moiety positioned C-terminally to the first polypeptide chain and/or the second polypeptide chain;
- wherein the half life-extending moiety is
- (i) an Fc polypeptide comprising any one of the amino acid sequences of SEQ ID NOS:
- 2-5 or a variant thereof containing no more than 10 amino acid substitutions, insertions, and/or deletions of a single amino acid per 100 amino acids;
- (ii) albumin or a fragment thereof;
- (iii) a fibronectin derivative comprising SEQ ID NO:
- 1, or a variant thereof containing no more than 10 amino acid substitutions, insertions, and/or deletions of a single amino acid per 100 amino acids, or a fragment thereof;
- or
- (iiii) polyethylene glycol (PEG);
- wherein the heterodimeric bispecific antibody binds to a cancer antigen on a target cell and a cluster of differentiation 3 (CD3) epsilon protein on a T cell and mediates cytolysis of the target cell by the T cell in vivo;
- wherein V1, V2, V3, and V4 are selected from the group consisting of:
- (a) V1 and V2 are each heavy chain variable regions and V3 and V4 are each light chain variable regions, and V1 and V3 create a complete VH/VL antigen binding pair binding the cancer antigen on the target cell and V2 and V4 create a complete VH/VL antigen binding pair binding the CD3 epsilon protein on the T cell;
- (b) V1 and V2 are each light chain variable regions and V3 and V4 are each heavy chain variable regions, and V1 and V3 create a complete VH/VL antigen binding pair binding the CD3 epsilon protein on the T cell and V2 and V4 create a complete VH/VL antigen binding pair binding the cancer antigen on the target cell;
- (c) V1 and V3 are each heavy chain variable regions and V2 and V4 are each light chain variable regions, and V1 and V4 create a complete VH/VL antigen binding pair binding the cancer antigen on the target cell and V2 and V3 create a complete VH/VL antigen binding pair binding the CD3 epsilon protein on the T cell;
- and
- (d) V1 and V3 are each light chain variable regions and V2 and V4 are each heavy chain variable regions, and V1 and V4 create a complete VH/VL antigen binding pair binding the CD3 epsilon protein on the T cell and V2 and V3 create a complete VH/VL antigen binding pair binding the cancer antigen on the target cell;
- and
- wherein L1 and L3 are each no more than 10 amino acids long;
- wherein when L2 and/or L4 are present, L2 and L4 are each no more than 10 amino acids long;
- wherein the VH region binding the CD3 epsilon protein comprises the heavy chain CDR1, CDR2, and CDR3 sequences in SEQ ID NO:
- 42, 44, or 82;
- and
- wherein the VL region binding the CD3 epsilon protein comprises the light chain CDR1, CDR2, and CDR3 sequences in SEQ ID NO:
- 43, 45, or 83.
26 more elements
A heterodimeric bispecific antibody comprising
- (a) a first polypeptide chain comprising an amino acid sequence having the formula V1-L1-V2-L2-CH1, wherein V1 and V2 are immunoglobulin variable regions, L1 and L2 are linkers, L2 can be present or absent, and CH1 is a first immunoglobulin heavy chain constant region comprising SEQ ID NO:
- 70;
- and
- (b) a second polypeptide chain comprising an amino acid sequence having the formula V3-L3-V4-L4-CL, wherein V3 and V4 are immunoglobulin variable regions, L3 and L4 are linkers, L4 can be present or absent, and CL is an immunoglobulin light chain constant region comprising SEQ ID NO:
- 71 or 73;
- wherein one of V1 and V4 is an immunoglobulin heavy chain variable (VH) region and the other is an immunoglobulin light chain variable (VL) region, and V1 and V4 can bind to a cancer antigen on a target cell or a cluster of differentiation 3 (CD3) epsilon protein on a T cell when they are part of an IgG and/or an scFv antibody;
- wherein one of V2 and V3 is a VH region and the other is a VL region, and V2 and V3 can bind to the cancer antigen on the target cell or the CD3 epsilon protein on the T cell when they are part of an IgG and/or an scFv antibody;
- wherein when V1 and V4 bind the cancer antigen V2 and V3 bind the CD3 epsilon protein, and when V1 and V4 bind the CD3 epsilon protein V2 and V3 bind the cancer antigen;
- wherein either or both of the first polypeptide chain and the second polypeptide chain further comprise(s) an in vivo half life-extending moiety positioned C-terminally to the first polypeptide chain and/or the second polypeptide-chain;
- wherein the half life-extending moiety is
- (i) an Fc polypeptide comprising any one of the amino acid sequences of SEQ ID NOS:
- 2-5 or a variant thereof containing no more than 10 amino acid substitutions, insertions, and/or deletions of a single amino acid per 100 amino acids;
- (ii) albumin or a fragment thereof;
- (iii) a fibronectin derivative comprising SEQ ID NO:
- 1, or a variant thereof containing no more than 10 amino acid substitutions, insertions, and/or deletions of a single amino acid per 100 amino acids, or a fragment thereof;
- or
- (iiii) polyethylene glycol (PEG);
- wherein the heterodimeric bispecific antibody binds to the cancer antigen and the CD3 epsilon protein and mediates cytolysis of the target cell by the T cell in vivo;
- wherein L1 and L3 are each no more than 10 amino acids long;
- wherein when L2 and/or L4 are present, L2 and L4 are each no more than 10 amino acids long;
- wherein the VH region binding the CD3 epsilon protein comprises the heavy chain CDR1, CDR2, and CDR3 sequences in SEQ ID NO:
- 42, 44, or 82;
- and
- wherein the VL region binding the CD3 epsilon protein comprises the light chain CDR1, CDR2, and CDR3 sequences in SEQ ID NO:
- 43, 45, or 83.
22 more elements
A heterodimeric bispecific antibody comprising
- (a) a first polypeptide chain comprising an amino acid sequence having the formula V1-L1-V2-CH1, wherein V1 and V2 are immunoglobulin variable regions, L1 is a linker no more than 10 amino acids long, and CH1 is a first immunoglobulin heavy chain constant region comprising SEQ ID NO:
- 70;
- and
- (b) a second polypeptide chain comprising an amino acid sequence having the formula V3-L3-V4-CL, wherein V3 and V4 are immunoglobulin variable regions, L3 is a linker no more than 10 amino acids long, and CL is an immunoglobulin light chain constant region comprising SEQ ID NO:
- 71 or 73;
- wherein either or both of the first polypeptide chain and the second polypeptide chain further comprise(s) an in vivo half life-extending moiety positioned C-terminally to the first polypeptide chain and/or the second polypeptide chain;
- wherein the half life-extending moiety is
- (i) an Fc polypeptide comprising any one of the amino acid sequences of SEQ ID NOS:
- 2-5 or a variant thereof containing no more than 10 amino acid substitutions, insertions, and/or deletions of a single amino acid per 100 amino acids;
- (ii) albumin or a fragment thereof;
- (iii) a fibronectin derivative comprising SEQ ID NO:
- 1, or a variant thereof containing no more than 10 amino acid substitutions, insertions, and/or deletions of a single amino acid per 100 amino acids, or a fragment thereof;
- or
- (iiii) polyethylene glycol (PEG);
- wherein the heterodimeric bispecific antibody binds to a cancer antigen on a target cell and a cluster of differentiation 3 (CD3) epsilon protein on a T cell and mediates cytolysis of the target cell by the T cell in vivo;
- wherein V1, V2, V3, and V4 are selected from the group consisting of:
- (a) V1 and V2 are each heavy chain variable regions and V3 and V4 are each light chain variable regions, and V1 and V3 create a complete VH/VL antigen binding pair binding the cancer antigen on the target cell and V2 and V4 create a complete VH/VL antigen binding pair binding the CD3 epsilon protein on the T cell;
- (b) V1 and V2 are each light chain variable regions and V3 and V4 are each heavy chain variable regions, and V1 and V3 create a complete VH/VL antigen binding pair binding the CD3 epsilon protein on the T cell and V2 and V4 create a complete VH/VL antigen binding pair binding the cancer antigen on the target cell;
- (c) V1 and V3 are each heavy chain variable regions and V2 and V4 are each light chain variable regions, and V1 and V4 create a complete VH/VL antigen binding pair binding the cancer antigen on the target cell and V2 and V3 create a complete VH/VL antigen binding pair binding the CD3 epsilon protein on the T cell;
- and
- (d) V1 and V3 are each light chain variable regions and V2 and V4 are each heavy chain variable regions, and V1 and V4 create a complete VH/VL antigen binding pair binding the CD3 epsilon protein on the T cell and V2 and V3 create a complete VH/VL antigen binding pair binding the cancer antigen on the target cell;
- and
- wherein the VH region binding the CD3 epsilon protein comprises the heavy chain CDR1, CDR2, and CDR3 sequences in SEQ ID NO:
- 42, 44, or 82;
- and
- wherein the VL region binding the CD3 epsilon protein comprises the light chain CDR1, CDR2, and CDR3 sequences in SEQ ID NO:
- 43, 45, or 83.
24 more elements
Dependencies are inferred from the claim text.