BISPECIFIC BINDING PROTEINS
The present disclosure provides bispecific proteins that bind to two antigens, as well as their compositions, uses, and methods of making.
1 . A bispecific binding protein capable of binding CD40 and capable of binding mesothelin, comprising two polypeptides of formula (II):
scFv X -H-Fc-L-scFv Y (II),
wherein
L is a polypeptide linker,
H is a hinge region,
Fc comprises CH2 and CH3 regions of an immunoglobulin,
scFv X and scFv Y are each independently a single chain variable fragment,
scFv X specifically binds a first antigen,
scFv Y specifically binds a second antigen, and
one of the two antigens is CD40 and the other is mesothelin.
2 . The bispecific binding protein of claim 1 , wherein scFv X - has the structure of formula (V) or (VI):
V H X -L X -V L X - (V),
V L X -L X -V H X - (VI),
wherein V H X is a variable heavy chain, V L X is a variable light chain, and L X is a polypeptide linker.
3 . The bispecific binding protein of claim 1 , wherein -scFv Y has the structure of formula (VII) or (VIII):
-V L Y -L Y -V H Y (VII),
-V H Y -L Y -V L Y (VIII),
wherein V H Y is a variable heavy chain, V L Y is a variable light chain, and L Y is a polypeptide linker.
4 . The bispecific binding protein of claim 1 , wherein L has an amino acid sequence comprising any one of SEQ ID NOS:251 and 253-262.
5 . The bispecific binding protein of claim 2 , wherein L X and L Y each has an amino acid sequence comprising any one of SEQ ID NOS:301-306.
6 . The bispecific binding protein of claim 1 , wherein the scFv that binds CD40 comprises a V H having a CDR-H1 according to SEQ ID NO:25, a CDR-H2 according to SEQ ID NO:26, and a CDR-H3 according to SEQ ID NO:27, and a V L having a CDR-L1 according to SEQ ID NO:55, a CDR-L2 according to SEQ ID NO:56, and a CDR-L3 according to SEQ ID NO:57.
7 . (canceled)
8 . The bispecific binding protein of claim 1 , wherein the scFv that binds CD40 comprises a V H according to SEQ ID NO: 17 and a V L according to SEQ ID NO:44.
9 .- 11 . (canceled)
12 . The bispecific binding protein of claim 1 , wherein the scFv that binds mesothelin comprises a V H having a CDR-H1 according to SEQ ID NO:241, a CDR-H2 according to SEQ ID NO:242, and a CDR-H3 according to SEQ ID NO:243, and a V L having a CDR-L1 according to SEQ ID NO:244, a CDR-L2 according to SEQ ID NO:245, and a CDR-L3 according to SEQ ID NO:246.
13 . (canceled)
14 . The bispecific binding protein of claim 1 , wherein the scFv that binds mesothelin comprises a V H according to SEQ ID NO:120 and a V L according to SEQ ID NO:137.
15 . The bispecific binding protein of claim 1 , wherein the scFv that binds mesothelin comprises a V H according to SEQ ID NO:120 and a V L according to SEQ ID NO:141.
16 .- 18 . (canceled)
19 . The bispecific binding protein of claim 1 , wherein the two polypeptides each comprise the amino acid sequence of SEQ ID NO:406.
20 . The bispecific binding protein of claim 1 , wherein the two polypeptides each comprise the amino acid sequence of SEQ ID NO:407.
21 . The bispecific binding protein of claim 1 , wherein the two polypeptides each comprise the amino acid sequence of SEQ ID NO:408.
22 . (canceled)
23 . The bispecific binding protein of claim 1 , wherein the two polypeptides each comprise the amino acid sequence of SEQ ID NO:419.
24 . The bispecific binding protein of claim 1 , wherein the two polypeptides each comprise the amino acid sequence of SEQ ID NO:420.
25 . (canceled)
26 . A pharmaceutical composition comprising a bispecific binding protein of any claim 1 , and a pharmaceutically acceptable carrier.
27 . A method of treating a cancer, comprising administering to a patient in need thereof a bispecific binding protein of claim 1 , optionally wherein the cancer is a solid tumor.
28 . A bispecific binding protein capable of binding 4-1BB and capable of binding prostate-specific membrane antigen, comprising two polypeptides of formula (II):
scFv X -H-Fc-L-scFv Y (II),
wherein
L is a polypeptide linker,
H is a hinge region,
Fc comprises CH2 and CH3 regions of an immunoglobulin,
scFv X and scFv Y are each independently a single chain variable fragment,
scFv X specifically binds a first antigen,
scFv Y specifically binds a second antigen, and
one of the two antigens is 4-1BB and the other is prostate-specific membrane antigen.
29 .- 32 . (canceled)
33 . The bispecific binding protein of claim 28 , wherein the scFv that binds 4-1BB comprises a V H having a CDR-H1 according to SEQ ID NO:280, a CDR-H2 according to SEQ ID NO:281, and a CDR-H3 according to SEQ ID NO:282, and a V L having a CDR-L1 according to SEQ ID NO:283, a CDR-L2 according to SEQ ID NO:284, and a CDR-L3 according to SEQ ID NO:285.
34 . (canceled)
35 . The bispecific binding protein of claim 33 , wherein the scFv that binds 4-1BB comprises a V H according to SEQ ID NO:69 and a V L according to SEQ ID NO:89.
36 . The bispecific binding protein of claim 33 , wherein the scFv that binds 4-1BB comprises a V H according to SEQ ID NO:71 and a V 1 according to SEQ ID NO:94.
37 . The bispecific binding protein of claim 28 , wherein the scFv that binds prostate-specific membrane antigen comprises a V H according to any one of SEQ ID NOS:191-197 and a V L according to any one of SEQ ID NOS:201-207.
38 .- 40 . (canceled)
41 . The bispecific binding protein of claim 28 , wherein the two polypeptides each comprise the amino acid sequence of SEQ ID NO:447.
42 . (canceled)
43 . A pharmaceutical composition comprising a bispecific binding protein of claim 28 , and a pharmaceutically acceptable carrier.
44 . A method of treating a cancer, comprising administering to a patient in need thereof a bispecific binding protein of claim 28 , optionally wherein the cancer is a solid tumor.
45 . A bispecific binding protein capable of binding CD40 and capable of binding mesothelin, comprising two polypeptides of formula (III):
Fab-H-Fc-L-scFv Y (III),
wherein
L is a polypeptide linker,
H is a hinge region,
Fc comprises CH2 and CH3 regions of an immunoglobulin,
Fab is a fragment antigen-binding region,
scFv Y is a single chain variable fragment,
Fab specifically binds a first antigen,
scFv Y specifically binds a second antigen, and
the first antigen is mesothelin and the second antigen is CD40.
46 .- 48 . (canceled)
49 . The bispecific binding protein of claim 45 , wherein the scFv that binds CD40 comprises a Vii having a CDR-H1 according to SEQ ID NO:25, a CDR-H2 according to SEQ ID NO:26, and a CDR-H3 according to SEQ ID NO:27, and a V L having a CDR-L1 according to SEQ ID NO:55, a CDR-L2 according to SEQ ID NO:56, and a CDR-L3 according to SEQ ID NO:57.
50 .- 51 . (canceled)
52 . The bispecific binding protein of claim 45 , wherein the scFv that binds CD40 comprises a Vii according to SEQ ID NO:22 and a V L according to SEQ ID NO:48.
53 . The bispecific binding protein of claim 45 , wherein the scFv that binds CD40 comprises a V H according to SEQ ID NO:23 and a V L according to SEQ ID NO:49.
54 . (canceled)
55 . The bispecific binding protein of claim 45 , wherein the Fab that binds mesothelin comprises a V H according to SEQ ID NO: 120 and a V L according to SEQ ID NO:137.
56 . The bispecific binding protein of claim 45 , wherein the Fab that binds mesothelin comprises a Vii according to SEQ ID NO:129 and a V L according to SEQ ID NO:143.
57 . (canceled)
58 . The bispecific binding protein of claim 45 , wherein the two polypeptides each comprise the heavy chain amino acid sequence of SEQ ID NO: 359 and the light chain amino acid sequence of SEQ ID NO: 371.
59 . The bispecific binding protein of claim 45 , wherein the two polypeptides each comprise the heavy chain amino acid sequence of SEQ ID NO: 360 and the light chain amino acid sequence of SEQ ID NO: 371.
60 .- 61 . (canceled)
62 . A pharmaceutical composition comprising a bispecific binding protein of claim 45 , and a pharmaceutically acceptable carrier.
63 . A method of treating a cancer, comprising administering to a patient in need thereof a bispecific binding protein of claim 45 , optionally wherein the cancer is a solid tumor.
64 . A bispecific binding protein comprising two polypeptides of formula (I):
X-H-Fc-L-scFv Y (I),
wherein
X is scFv X or a Fab region,
wherein X specifically binds a first antigen and scFv Y specifically binds a second antigen,
H is a hinge region,
Fc comprises CH2 and CH3 regions of an immunoglobulin,
L is a polypeptide linker,
scFv X and scFv Y are each independently a single chain variable fragment,
one of the two antigens is an immunomodulatory protein and the other is a tumor antigen, and
the bispecific binding protein activates the immunomodulatory protein.
65 . (canceled)
66 . The bispecific binding protein of claim 64 , wherein the immunomodulatory protein is selected from CD40, 4-1BB, TNFR2, ICOS, TRAILR1, and TRAILR2.
67 .- 68 . (canceled)
69 . The bispecific binding protein of claim 64 , wherein the tumor antigen is selected from mesothelin, nectin-4, epidermal growth factor receptor, prostate-specific membrane antigen, and B7-H4.
70 . (canceled)
71 . The bispecific binding protein of claim 64 , wherein the first antigen is an immunomodulatory protein and the second antigen is a tumor antigen.
72 . (canceled)
73 . A pharmaceutical composition comprising a bispecific binding protein of claim 64 , and a pharmaceutically acceptable carrier.
74 . (canceled)
75 . A nucleic acid comprising a nucleotide sequence encoding a bispecific binding protein of claim 1 .
76 .- 77 . (canceled)
78 . A eukaryotic host cell transformed with the vector comprising the nucleic acid of claim 75 .
79 .- 80 . (canceled)
81 . A method of producing a bispecific binding protein, comprising: (a) culturing the host cell of claim 78 and (b) recovering the protein.
82 . A method of activating the immune system, comprising administering to a patient in need thereof a bispecific binding protein of claim 1 .