# US 12,215,166 B2 · Bispecific antigen binding constructs targeting HER2 > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,215,166 B2 - Document ID: US12215166B2 - Status: Granted - Kind: B2 - Application number: 17/728,491 - Filed: 2022-04-25 - Published: 2023-04-20 - Granted: 2025-02-04 - Earliest claimed: 2013-11-27 - Assignees: Zymeworks BC Inc. - Applicants: Zymeworks Bc Inc. - Inventors: Nina E. Weisser; Gordon Yiu Kon Ng; Grant Raymond Wickman; Surjit Bhimarao Dixit; Eric Escobar-Cabrera; Mario Sanches - Web page: https://patentgrep.com/patent/US12215166B2 ## CPC - C07K 16/32: against translation products of oncogenes - A61K 39/39558: {against tumor tissues, cells, antigens} - A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca - A61K 47/68033: {the drug being a maytansine} - A61K 47/6851: {the antibody targeting a determinant of a tumour cell} - A61K 47/6855: {the tumour determinant being from breast cancer cell} - A61K 47/6869: {the tumour determinant being from a cell of the reproductive system: ovaria, uterus, testes, prostate} - A61P 35/00: Antineoplastic agents - C07K 16/3015: {Breast} - C07K 16/3069: {Reproductive system, e.g. ovaria, uterus, testes, prostate} - G01N 33/5759: {involving compounds localised on the membrane of tumour or cancer cells} - A61K 2039/505: {comprising antibodies} - C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered - C07K 2317/31: multispecific - C07K 2317/35: Valency - C07K 2317/41: Glycosylation, sialylation, or fucosylation - C07K 2317/52: Constant or Fc region; Isotype - C07K 2317/524: CH2 domain - C07K 2317/526: CH3 domain - C07K 2317/55: Fab or Fab' - C07K 2317/622: Single chain antibody (scFv) - C07K 2317/73: Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation - C07K 2317/732: Antibody-dependent cellular cytotoxicity [ADCC] - C07K 2317/77: Internalization into the cell - C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value - C07K 2317/94: Stability, e.g. half-life, pH, temperature or enzyme-resistance - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract 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. ## Claims ### Claim 1 An isolated antigen binding construct comprising: (a) 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 the sequence shown in SEQ ID NO:99 and the VL1 comprises the sequence shown in SEQ ID NO:71; and (b) a second antigen binding polypeptide construct which specifically binds a HER2 ECD4 (extracellular domain 4) antigen on a HER2-expressing cell, wherein the second antigen binding polypeptide construct comprises a second heavy chain variable domain (VH2) and a second light chain variable domain (VL2), wherein the VH2 comprises the sequence shown in SEQ ID NO: 305 and the VL2 comprises the sequence shown in SEQ ID NO:297, and wherein the second antigen binding polypeptide construct is operably linked to the first antigen binding polypeptide construct. ### Claim 2 (depends on 1) The isolated construct of claim 1, wherein the first antigen binding polypeptide is a Fab and the second antigen binding polypeptide is a scFv. ### Claim 3 (depends on 1) The isolated construct of claim 1, wherein the first antigen binding polypeptide is an scFv and the second antigen binding polypeptide is a Fab. ### Claim 4 (depends on 2) The isolated construct of claim 2, 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 comprises the sequence shown in SEQ ID NO:97 and the L1 comprises the sequence shown in SEQ ID NO:69; and the second antigen binding polypeptide construct comprises the sequence shown in SEQ ID NO:295. ### Claim 5 (depends on 1) The isolated construct of claim 1, further comprising a heterodimeric Fc, wherein the heterodimeric Fc is a human IgG1 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; and wherein 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 wild-type homodimeric human IgG1 Fc. ### Claim 6 An isolated polynucleotide or set of isolated polynucleotides that encode(s): (a) 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 the sequence shown in SEQ ID NO:99 and the VL1 comprises the sequence shown in SEQ ID NO:71; and (b) a second antigen binding polypeptide construct which specifically binds a HER2 ECD4 (extracellular domain 4) antigen on a HER2-expressing cell, wherein the second antigen binding polypeptide construct comprises a second heavy chain variable domain (VH2) and a second light chain variable domain (VL2), wherein the VH2 comprises the sequence shown in SEQ ID NO:305 and the VL2 comprises the sequence shown in SEQ ID NO:297. ### Claim 7 (depends on 6) The isolated polynucleotide or set of isolated polynucleotides of claim 6, 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 comprises the sequence shown in SEQ ID NO:97 and the L1 comprises the sequence shown in SEQ ID NO:69. ### Claim 8 (depends on 6) The isolated polynucleotide or set of isolated polynucleotides of claim 6, wherein the second antigen binding polypeptide construct comprises the sequence shown in SEQ ID NO:295. ### Claim 9 (depends on 7) The isolated polynucleotide or set of isolated polynucleotides of claim 7, wherein the second antigen binding polypeptide construct comprises the sequence shown in SEQ ID NO:295. ### Claim 10 (depends on 6) The isolated polynucleotide or set of isolated polynucleotides of claim 6, further encoding a heterodimeric Fc, wherein the heterodimeric Fc is a human IgG1 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 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 wild-type homodimeric human IgG1 Fc. ### Claim 11 (depends on 9) The isolated polynucleotide or set of isolated polynucleotides of claim 9, further encoding a heterodimeric Fc, wherein the heterodimeric Fc is a human IgG1 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 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 wild-type homodimeric human IgG1 Fc. ### Claim 12 A vector or set of vectors comprising an isolated polynucleotide or set of isolated polynucleotides that encode(s): (a) 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 the sequence shown in SEQ ID NO:99 and the VL1 comprises the sequence shown in SEQ ID NO:71; and (b) a second antigen binding polypeptide construct which specifically binds a HER2 ECD4 (extracellular domain 4) antigen on a HER2-expressing cell, wherein the second antigen binding polypeptide construct comprises a second heavy chain variable domain (VH2) and a second light chain variable domain (VL2), wherein the VH2 comprises the sequence shown in SEQ ID NO:305 and the VL2 comprises the sequence shown in SEQ ID NO:297. ### Claim 13 (depends on 12) The vector or set of vectors of claim 12, 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 comprises the sequence shown in SEQ ID NO:97 and the L1 comprises the sequence shown in SEQ ID NO:69. ### Claim 14 (depends on 12) The vector or set of vectors of claim 12, wherein the second antigen binding polypeptide construct comprises the sequence shown in SEQ ID NO:295. ### Claim 15 (depends on 13) The vector or set of vectors of claim 13, wherein the second antigen binding polypeptide construct comprises the sequence shown in SEQ ID NO:295. ### Claim 16 (depends on 12) The vector or set of vectors of claim 12, where the isolated polynucleotide or set of isolated polynucleotides further encode(s) a heterodimeric Fc, wherein the heterodimeric Fc is a human IgG1 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 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 wild-type homodimeric human IgG1 Fc. ### Claim 17 (depends on 15) The vector or set of vectors of claim 15, where the isolated polynucleotide or set of isolated polynucleotides further encode(s) a heterodimeric Fc, wherein the heterodimeric Fc is a human IgG1 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 CH3 sequence comprises T350V_L351Y_F405A_Y407V in the first Fc polypeptide, and the second CH3 sequence comprises T350V_T366L_K392L_T394W. ### Claim 18 (depends on 6) An isolated cell comprising the isolated polynucleotide or set of isolated polynucleotides of claim 6. ### Claim 19 (depends on 7) An isolated cell comprising the isolated polynucleotide or set of isolated polynucleotides of claim 7. ### Claim 20 (depends on 8) An isolated cell comprising the isolated polynucleotide or set of isolated polynucleotides of claim 8. ### Claim 21 (depends on 9) An isolated cell comprising the isolated polynucleotide or set of isolated polynucleotides of claim 9. ### Claim 22 (depends on 10) An isolated cell comprising the isolated polynucleotide or set of isolated polynucleotides of claim 10. ### Claim 23 (depends on 11) An isolated cell comprising the isolated polynucleotide or set of isolated polynucleotides of claim 11.