Bispecific antigen-binding constructs targeting HER2
Provided herein are biparatopic antigen-binding constructs that specifically bind HER2. The biparatopic antigen-binding constructs comprise one antigen-binding moiety that binds to ECD2 of HER2, a second antigen-binding moiety that binds to ECD4 of HER2, and an Fc. At least one of the antigen-binding moieties is an scFv. The biparatopic antigen-binding constructs can be used in the treatment of cancer.
1. A method of treating a subject having a HER2 (human epidermal growth factor receptor 2) expressing (HER2+) tumor comprising administering an effective amount of an antigen binding construct to the subject, wherein the antigen binding construct comprises:
a first antigen-binding polypeptide construct which specifically binds a HER2 (human epidermal growth factor receptor 2) ECD2 (extracellular domain 2) antigen on a HER2-expressing cell, wherein the first antigen binding polypeptide construct comprises a first heavy chain variable region (VH1) and a first light chain variable region (VL1), wherein the VH1 comprises a first complementarity determining region (CDR1) comprising the amino acid sequence shown in SEQ ID NO:348, a second complementarity determining region (CDR2) comprising the amino acid sequence shown in SEQ ID NO:335, and a third complementarity determining region (CDR3) comprising the amino acid sequence shown in SEQ ID NO:336, and the VL1 comprises a CDR1 comprising the amino acid sequence shown in SEQ ID NO:340, a CDR2 comprising the amino acid sequence shown in SEQ ID NO:338, and a CDR3 comprising the amino acid sequence shown in SEQ ID NO:347, wherein the first antigen-binding polypeptide construct comprises a first heavy chain (H1) comprising the VH1 and a first light chain (L1) comprising the VL1, and the first H1 comprises the amino acid sequence shown in SEQ ID NO:97 and the first L1 comprises the amino acid sequence shown in SEQ ID NO:69;
wherein the antigen binding construct further comprises a heterodimeric Fc comprising a first Fc polypeptide and a second Fc polypeptide, the first Fc polypeptide comprising a first CH3 sequence and the second Fc polypeptide comprising a second CH3 sequence, wherein the first Fc polypeptide is operably linked to the first antigen-binding polypeptide construct.
2. The method of claim 1 , wherein the first antigen binding polypeptide construct is a Fab.
3. The method of claim 1 , wherein the first H1 consists of the amino acid sequence shown in SEQ ID NO:97 and the first L1 consists of the amino acid sequence shown in SEQ ID NO:69.
4. The method of claim 1 , wherein the antigen binding construct further comprises a second antigen binding polypeptide construct operably linked to the first antigen binding polypeptide construct, wherein the second antigen binding polypeptide construct comprises a second heavy chain variable domain (VH2) and a second light chain variable domain (VL2).
5. The method of claim 4 , wherein the VH2 comprises a CDR1 comprising the amino acid sequence shown in SEQ ID NO:343, a CDR2 comprising the amino acid sequence shown in SEQ ID NO:341, and a CDR3 comprising the amino acid sequence shown in SEQ ID NO:342, and the VL2 comprises a CDR1 comprising the amino acid sequence shown in SEQ ID NO:346, a CDR2 comprising the amino acid sequence shown in SEQ ID NO:344, and a CDR3 comprising the amino acid sequence shown in SEQ ID NO:345.
6. The method of claim 4 , wherein the first antigen polypeptide construct is a Fab and the second antigen polypeptide construct is an scFv.
7. The method of claim 5 , wherein the first antigen polypeptide construct is a Fab and the second antigen polypeptide construct is an scFv.
8. The method of claim 5 , wherein the first antigen binding polypeptide construct is a Fab and the VH1 comprises the amino acid sequence shown in SEQ ID NO:99 and the VL1 comprises the amino acid sequence shown in SEQ ID NO:71.
9. The method of claim 5 , wherein the first antigen binding polypeptide construct is a Fab and the VH1 comprises the amino acid sequence shown in SEQ ID NO:99 and the VL1 comprises the amino acid sequence shown in SEQ ID NO:71; and the second antigen binding polypeptide construct is an scFv and the VH2 comprises the amino acid sequence shown in SEQ ID NO:305 and the VL2 comprises the amino acid sequence shown in SEQ ID NO:297.
10. The method of claim 5 , wherein the first antigen binding polypeptide construct comprises a first heavy chain (H1) comprising the VH1 and a first light chain (L1) comprising the VL1, and the H1 consists of the amino acid sequence shown in SEQ ID NO:97 and the L1 consists of the amino acid sequence shown in SEQ ID NO:69; and the second antigen binding polypeptide construct consists of the amino acid sequence shown in SEQ ID NO:295.
11. The method of claim 1 , wherein the heterodimeric Fc is a human IgG1 Fc, the first Fc polypeptide is operably linked to the first antigen binding polypeptide construct with an IgG1 hinge region linker, the first CH3 sequence comprises T350V_L351Y_F405A_Y407V in the first Fc polypeptide, and the second CH3 sequence comprises T350V_T366L_K392L_T394W according to EU numbering compared to a wide-type homodimeric human IgG1 Fc.
12. The method of claim 5 , wherein the antigen binding construct further comprises a heterodimeric Fc comprising a first Fc polypeptide and a second Fc polypeptide, the first Fc polypeptide comprising a first CH3 sequence and the second Fc polypeptide comprising a second CH3 sequence, wherein the first Fc polypeptide is operably linked to the first antigen-binding polypeptide construct and the second Fc polypeptide is operably linked to the second antigen-binding polypeptide construct.
13. The method of claim 12 , wherein the heterodimeric Fc is a human IgG1 Fc, the first Fc polypeptide is operably linked to the first antigen binding polypeptide construct with an IgG1 hinge region linker, the first CH3 sequence comprises T350V_L351Y_F405A_Y407V modifications in the first Fc polypeptide, and the second CH3 sequence comprises T350V_T366L_K392L_T394W modifications according to EU numbering compared to a wild-type homodimeric human IgG1 Fc.
14. The method of claim 12 , where the heterodimeric Fc is a human IgG1 Fc, the first Fc polypeptide is operably linked to the first antigen binding polypeptide construct with an IgG1 hinge region linker, the first CH3 sequence comprises T350V_L351Y_F405A_Y407V modifications in the first Fc polypeptide, the second Fc polypeptide is operably linked to the second antigen binding polypeptide construct with an IgG1 hinge region linker, and the second CH3 sequence comprises T350V_T366L_K392L_T394W modifications according to EU numbering compared to a wild-type homodimeric human IgG1 Fc.
15. The method of claim 1 , wherein the heterodimeric Fc comprises one or more CH2 modifications.
16. The method of claim 1 , wherein the antigen binding construct is glycosylated.
17. The method of claim 1 , wherein the antigen binding construct is conjugated to a drug.
18. A method of treating a subject having a HER2 (human epidermal growth factor receptor 2) expressing (HER2+) tumor comprising administering an effective amount of antigen binding construct to the subject, wherein the antigen binding construct comprises:
a heavy chain 1 (H1), a heavy chain 2 (H2), and a light chain 1 (L1), wherein:
(a) H1 comprises the complementarity determining region (CDR) sequences CDR1, CDR3, and CDR2 set forth in SEQ ID NO: 101, SEQ ID NO: 103, and SEQ ID NO: 105, respectively;
(b) H2 comprises the CDR sequences CDR-L1, CDR-L3, and CDR-L2 set forth in SEQ ID NO:
299, SEQ ID NO: 301, SEQ ID NO: 303, respectively, and the CDR sequences CDR-H1, CDR-H3 and CDR-H2 set forth in SEQ ID NO: 307, SEQ ID NO: 309, and SEQ ID NO: 311, respectively; and
(c) L1 comprises the CDR sequences CDR1, CDR3 and CDR2 set forth in SEQ ID NO: 73, SEQ ID NO: 75, and SEQ ID NO: 77 respectively
wherein the antigen binding construct further comprises a heterodimeric Fc comprising a first Fc polypeptide and a second Fc polypeptide, the first Fc polypeptide comprising a first CH3 sequence and the second Fc polypeptide comprising a second CH3 sequence, wherein the first Fc polypeptide is operably linked to the first antigen-binding polypeptide construct.
19. The method of claim 18 , wherein (a) H1 comprises the amino acid sequence set forth in SEQ ID NO: 97; (b) H2 comprises the amino acid sequence set forth in SEQ ID NO: 295; and (c) L1 comprises the amino acid sequence set forth in SEQ ID NO: 69.