CD3-targeting antibody, bispecific antibody and use thereof
Disclosed are a CD3-targeting antibody, a bispecific antibody and the use thereof. The CD3-targeting antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH). The VL has the amino acid sequence as shown in SEQ ID NO: 56 or a mutation thereof. The VH has mutations on the amino acid sequence as shown in SEQ ID NO: 42, and the mutations occur at one or more of the sites of amino acid residues selected from positions 30, 73, 76, 78, 93 and 94. The bispecific antibody comprises a first protein functional region and a second protein functional region, wherein the first protein functional region comprises the CD3-targeting antibody as described above. The CD3-targeting antibody reduces the toxicity caused by cytokine release syndrome, and the bispecific antibody prepared therefrom is stable and has the ability to bind to T cells, and also reduces the difficulty of producing.
1 . A CD3-targeting antibody, comprising a light chain variable region (VL) and a heavy chain variable region (VH), wherein,
the amino acid sequence of the VH is set forth in SEQ ID NO: 44, and the amino acid sequence of the VL is set forth in SEQ ID NO: 58; or,
the amino acid sequence of the VH is set forth in SEQ ID NO: 51, and the amino acid sequence of the VL is set forth in SEQ ID NO: 58; or,
the amino acid sequence of the VH is set forth in SEQ ID NO: 45, and the amino acid sequence of the VL is set forth in SEQ ID NO: 58; or,
the amino acid sequence of the VH is set forth in SEQ ID NO: 52, and the amino acid sequence of the VL is set forth in SEQ ID NO: 58; or,
the amino acid sequence of the VH is set forth in SEQ ID NO: 43, and the amino acid sequence of the VL is set forth in SEQ ID NO: 58; or,
the amino acid sequence of the VH is set forth in SEQ ID NO: 50, and the amino acid sequence of the VL is set forth in SEQ ID NO: 58; or,
the amino acid sequence of the VH is set forth in SEQ ID NO: 47, and the amino acid sequence of the VL is set forth in SEQ ID NO: 58; or,
the amino acid sequence of the VH is set forth in SEQ ID NO: 48, and the amino acid sequence of the VL is set forth in SEQ ID NO: 58; or,
the amino acid sequence of the VH is set forth in SEQ ID NO: 49, and the amino acid sequence of the VL is set forth in SEQ ID NO: 58; or,
the amino acid sequence of the VH is set forth in SEQ ID NO: 53, and the amino acid sequence of the VL is set forth in SEQ ID NO: 58; or,
the amino acid sequence of the VH is set forth in SEQ ID NO: 54, and the amino acid sequence of the VL is set forth in SEQ ID NO: 58; or,
the amino acid sequence of the VH is set forth in SEQ ID NO: 55, and the amino acid sequence of the VL is set forth in SEQ ID NO: 58; or,
the amino acid sequence of the VH is set forth in SEQ ID NO: 46 and the amino acid sequence of the VL is set forth in SEQ ID NO: 58.
2 . The CD3-targeting antibody of claim 1 , wherein the antibody comprises a single chain antibody (scFv) of VL linker-VH or VH-linker-VL.
3 . The CD3-targeting antibody of claim 1 , wherein the antibody further comprises a constant region.
4 . A bispecific antibody, comprising a first protein functional region and a second protein functional region, wherein the first protein functional region comprises the CD3-targeting antibody of claim 1 , and the second protein functional region is a ROR1 targeting antibody.
5 . The bispecific antibody of claim 4 , wherein the bispecific antibody comprises the following three chains: (1) VL1-linker-VH1-Hinge-CH2-CH3 (knob) or VH1-linker-VL1-Hinge-CH2-CH3 (knob) of the first protein functional region, (2) VH2-CH1-Hinge-CH2-CH3 (hole) of the second protein functional region, and (3) VL2-CL of the second protein functional region; the second protein functional region is a non-CD3-targeting antibody.
6 . The bispecific antibody of claim 5 , wherein the bispecific antibody comprises VL1-linker-VH1-Hinge-CH2-CH3 (knob) as shown in SEQ ID NO: 88, VH2-CH1-Hinge-CH2-CH3 (hole) as shown in SEQ ID NO: 87, and VL2-CL as shown in SEQ ID NO: 85.
7 . An isolated nucleic acid, encoding the CD3-targeting antibody of claim 1 or a bispecific antibody comprising a first protein functional region and a second protein functional region, wherein the first protein functional region comprises the CD3-targeting antibody, and the second protein functional region is a ROR1 targeting antibody.
8 . An expression vector, comprising the isolated nucleic acid of claim 7 .
9 . A genetically modified cell, transfected with the expression vector of claim 8 .
10 . A pharmaceutical composition, comprising the CD3-targeting antibody of claim 1 , a bispecific antibody, and a pharmaceutically acceptable carrier;
wherein the bispecific antibody comprises a first protein functional region and a second protein functional region, wherein the first protein functional region comprises the CD3-targeting antibody, and the second protein functional region is a ROR1 targeting antibody.
11 . A method for treating a patient in need of a medicament for tumor, comprising administering to the patient a medicament comprising an effective amount of the CD3-targeting antibody of claim 1 , a bispecific antibody or a pharmaceutical composition;
wherein the bispecific antibody comprises a first protein functional region and a second protein functional region, wherein the first protein functional region comprises the CD3-targeting antibody, and the second protein functional region is a ROR1 targeting antibody;
wherein the pharmaceutical composition comprises the CD3-targeting antibody, the bispecific antibody and a pharmaceutically acceptable carrier.
12 . The CD3-targeting antibody of claim 2 , wherein the linker is (G 4 S) n or a variant thereof, wherein n is a non-zero natural number.
13 . The CD3-targeting antibody of claim 12 , wherein the amino acid sequence of the scFv is set forth in SEQ ID NO: 73, SEQ ID NO: 78, SEQ ID NO: 79 or SEQ ID NO: 80.
14 . The CD3-targeting antibody of claim 13 , wherein the antibody further comprises the Fc, wherein the Fc is linked to the scFv by a hinge.
15 . The CD3-targeting antibody of claim 3 , wherein the constant region is a human constant region comprising a human light chain constant region and a human heavy chain constant region.
16 . The CD3-targeting antibody of claim 15 , wherein the human heavy chain constant region is from hIgG1, hIgG2, hIgG3, hIgG4, or a variant thereof.
17 . The bispecific antibody of claim 5 , wherein the linker is (G 4 S) n , and n is a non-zero natural number of 1 to 20.
18 . The CD3-targeting antibody of claim 16 , wherein the human heavy chain constant region is a heavy chain constant region as shown in SEQ ID NO: 63 or SEQ ID NO: 64.