Multi-specific antibodies and methods of using thereof
The application provides tri-specific antibody monomers having a N-terminal and a C-terminal, comprising in tandem from the N-terminal to the C-terminal, a first scFv domain at the N-terminal, a Fab domain, a Fc domain, and a second scFv domain at the C-terminal. In one embodiment, the first scFv domain, the Fab domain, and the second scFv domain each has a binding specificity against a different antigen.
1 . A method for treating a cancer in a subject, said method comprising administering to the subject an effective amount of a pharmaceutical composition comprising a purified multi-specific antibody, wherein the multi-specific antibody comprises a tri-specific antibody monomer having a N-terminal and a C-terminal, comprising in tandem from the N-terminal to the C-terminal,
a first scFv domain at the N-terminal, comprising the following pairs of amino acid sequences:
SEQ ID NOs: (2 and 4), (6 and 8), or (10 and 12), having the binding specificity against ROR1,
SEQ ID NOs: (14 and 16), having the binding specificity against PD-L1,
SEQ ID NOs: (40 and 42), having the binding specificity against EGFRvIII, or
SEQ ID NOs: (44 and 46), having the binding specificity against CD19,
a Fab domain, having a binding specificity against PD-L1, EGFRvIII, or CD19,
a Fc domain, and
a second scFv domain at the C-terminal, having a binding specificity against CD3 and comprising the following pairs of sequences SEQ ID NOs: (18 and 20), (22 and 24), (26 and 28), (30 and 32), or (30 and 34),
wherein the first scFv domain, the Fab domain, and the second scFv domain each has a binding specificity against a different antigen, and wherein the cancer comprises cells expressing ROR1, PD-L1, EGFRvIII, or CD19.
2 . The method of claim 1 , further comprising co-administering an effective amount of a therapeutic agent.
3 . The method of claim 2 , wherein the therapeutic agent comprises an anti-estrogen agent, a receptor tyrosine kinase inhibitor, a kinase inhibitor, a cell cycle inhibitor, a DNA, RNA or protein synthesis inhibitor, a RAS inhibitor, a check point inhibitor, an inhibitor of PD1, PDL1, CTLA4, 4-1BB, OX40, GITR, ICOS, LIGHT, TIM3, LAG3, TIGIT, CD40, CD27, HVEM, BTLA, VISTA, B7H4, CSF1R, NKG2D, CD73, a derivative or a combination thereof.
4 . The method of claim 1 , wherein the first scFv domain the has a binding specificity against EGFRvIII and comprises 3 CDRs of SEQ ID NO:40 and 3 CDRs of SEQ ID NO:42,
wherein the first scFv domain has a binding specificity against CD19 and comprises 3 CDRs of SEQ ID NO:44 and 3 CDRs of SEQ ID NO:46, or
wherein the first scFv domain has a binding specificity against PD-L1 and comprises 3 CDRs of SEQ ID NO: 14 and 3 CDRs of SEQ ID NO: 16.
5 . The method of claim 1 , wherein the first scFv domain, the Fab domain, and the second scFv domain each independently has a binding specificity against an antigen selected from ROR1, PD-L1, and CD3.
6 . The method of claim 1 , wherein the first scFv domain has a binding specificity against ROR1, and wherein the first scFv comprises 3 complementarity determining regions (CDRs) of SEQ ID NO:6 and 3 CDRs of SEQ ID NO:8.
7 . The method of claim 1 , wherein the Fab domain has a binding specificity against PD-L1.
8 . The method of claim 1 , wherein the second scFv domain has a binding specificity against CD3, and wherein the second scFv comprises 3 CDRs of SEQ ID NO: 22 and 3 CDRs of SEQ ID NO:24.
9 . The method of claim 1 , wherein the first scFv domain has a binding specificity against PD-L1, and wherein the first scFv comprises 3 CDRs of SEQ ID NO: 14 and 3 CDRs of SEQ ID NO: 16.
10 . The method of claim 1 , wherein the Fc domain comprises SEQ ID NO: 48 or 50.
11 . The method of claim 1 , wherein the tri-specific antibody monomer has a binding specificity to human PD-L1, human CD3, and one of human ROR1, human CD19, or EGFR VIII.
12 . The method of claim 1 , wherein the first scFv domain comprises a (G4S) n linker, wherein n is an integral from 2 to 4 and wherein the first scFv domain is linked to the Fab domain through the linker.
13 . The method of claim 1 , wherein the tri-specific antibody monomer comprises a binding domain having a binding specificity to human ROR1, wherein the binding domain comprises a pair of amino acid sequences selected from SEQ ID: 2 and 4, SEQ ID: 6 and 8, and SEQ ID: 10 and 12.
14 . The method of claim 1 , wherein the tri-specific antibody monomer comprises a binding domain having a binding specificity to human CD19, wherein the binding domain comprises a pair of amino acid sequences of SEQ ID: 44 and 46.
15 . The method of claim 1 , wherein the tri-specific antibody monomer comprises a binding domain having a binding specificity to EGFR vIII, wherein the binding domain comprises a pair of amino acid sequences of SEQ ID: 40 and 42.
16 . The method of claim 1 , wherein the tri-specific antibody monomer comprises a binding domain having a binding specificity to human PD-L1, wherein the binding domain comprises a pair of amino acid sequences of SEQ ID: 14 and 16.
17 . A method for treating a cancer that comprises cells expressing ROR1, PD-L1, EGFRvIII, or CD19 in a subject, said method comprising administering to the subject an effective amount of a pharmaceutical composition comprising a tri-specific antibody having a N-terminal and a C-terminal, comprising in tandem from the N-terminal to the C-terminal,
a first scFv domain at the N-terminal,
a Fab domain,
a Fc domain, and
a second scFv domain at the C-terminal, comprising 3 CDRs of SEQ ID NO: 22 and 3 CDRs of SEQ ID NO: 24,
wherein the tri-specific antibody comprises 3 CDRs of SEQ ID NO: 14 and 3 CDRs of SEQ ID NO: 16, and one of:
3 CDRs of SEQ ID NO: 6 and 3 CDRs of SEQ ID NO: 8,
3 CDRs of SEQ ID NO: 40 and 3 CDRs of SEQ ID NO: 42, and
3 CDRs of SEQ ID NO: 44 and 3 CDRs of SEQ ID NO: 46.
18 . The method of claim 17 , wherein the tri-specific antibody has a binding specificity to PD-L1, CD3, and one of ROR1, CD19 and EGFRvIII.