Methods of treating cancer by administering antigen-binding molecules comprising a TNF family ligand trimer

Patent metadata

Field	Value
Publication number	US-11306154-B2
Application number	US-201916522391-A
Country	US
Kind code	B2
Filing date	Jul 25, 2019
Priority date	Nov 14, 2014
Publication date	Apr 19, 2022
Grant date	Apr 19, 2022

Abstract

The invention relates to novel TNF family ligand trimer-containing antigen binding molecules comprising (a) at least one moiety capable of specific binding to a target cell antigen and (b) a first and a second polypeptide that are linked to each other by a disulfide bond, characterized in that the first polypeptide comprises two ectodomains of a TNF ligand family member or fragments thereof that are connected to each other by a peptide linker and in that the second polypeptide comprises only one ectodomain of said TNF ligand family member or a fragment thereof.

First claim

What is claimed is:

1. A method of treating a cancer in an individual, the method comprising administering to said individual a therapeutically effective amount of a composition comprising a TNF family ligand trimer-containing antigen-binding molecule, said molecule comprising:
(a) at least one antigen-binding domain comprising a light chain variable region (VL) and a heavy chain variable region (VH) capable of specific binding to a target cell antigen, wherein the target cell antigen is Fibroblast Activation Protein (FAP),
(b) a first polypeptide and a second polypeptide that are linked to each other by a disulfide bond, wherein the first polypeptide comprises two ectodomains of a TNF family ligand, or fragments thereof, that are connected to each other by a peptide linker and the second polypeptide comprises only one ectodomain of the TNF family ligand, or fragment thereof,
wherein the ectodomain of the TNF family ligand, or fragment thereof, comprises the amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO: 2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:96, SEQ ID NO: 373, SEQ ID NO:374, and SEQ ID NO:375, and
(c) an Fc domain composed of a first subunit and a second subunit, wherein the first subunit and the second subunit are capable of stable association with each other,
wherein the TNF family ligand is 4-1BBL, and wherein the composition comprising the TNF family ligand trimer-containing antigen-binding molecule is in a pharmaceutically acceptable form.

2. The method of claim 1 , wherein the cancer is a solid tumor, bladder cancer, renal cell carcinoma, brain cancer, head and neck cancer, pancreatic cancer, lung cancer, breast cancer, ovarian cancer, uterine cancer, cervical cancer, endometrial cancer, esophageal cancer, colon cancer, colorectal cancer, rectal cancer, gastric cancer, prostate cancer, blood cancer, skin cancer, squamous cell carcinoma, bone cancer, kidney cancer, melanoma, B-cell lymphoma, B-cell leukemia, non-Hodgkin lymphoma, or acute lymphoblastic leukemia.

3. The method of claim 1 , wherein
(i) the first polypeptide comprises a heavy chain constant 1 (CH1) domain or a light chain constant (CL) domain and the second polypeptide comprises a CL domain or a CH1 domain, wherein the second polypeptide is linked to the first polypeptide by a disulfide bond between the CH1 domain and the CL domain, wherein the first polypeptide comprises two ectodomains of the TNF family ligand, or fragments thereof, that are connected to each other and to the CH1 domain or the CL domain of the first polypeptide by a peptide linker, and wherein the second polypeptide comprises only one ectodomain of the TNF family ligand, or fragment thereof, connected to the CL domain or the CH1 domain of the second polypeptide by a peptide linker,
(ii) the first polypeptide comprises a heavy chain constant 3 (CH3) domain and the second polypeptide comprises a CH3 domain, wherein the first polypeptide comprises two ectodomains of the TNF family ligand, or fragments thereof, that are connected to each other and to the C-terminus of the CH3 domain of the first polypeptide by a peptide linker, and wherein the second polypeptide comprises only one ectodomain of the TNF family ligand, or fragment thereof, connected to C-terminus of the CH3 domain of the second polypeptide by a peptide linker, or
(iii) the first polypeptide comprises a heavy chain variable region-light chain constant domain (VH-CL) or a light chain variable region-heavy chain constant 1 domain (VL-CH1) and the second polypeptide comprises a VL-CH1 domain or a VH-CL domain, wherein the second polypeptide is linked to the first polypeptide by a disulfide bond between the CH1 domain and the CL domain, wherein the first polypeptide comprises two ectodomains of the TNF family ligand, or fragments thereof, that are connected to each other and to the VH or the VL of the first polypeptide by a peptide linker, and wherein the second polypeptide comprises only one ectodomain of the TNF family ligand, or fragment thereof, connected to the VL or the VH of the second polypeptide by a peptide linker.

4. The method of claim 1 , wherein the TNF family ligand costimulates human T-cell activation.

5. The method of claim 1 , wherein the ectodomain of the TNF family ligand, or fragment thereof, comprises the amino acid sequence of SEQ ID NO:1.

6. The method of claim 1 , wherein the ectodomain of the TNF family ligand, or fragment thereof, comprises the amino acid sequence of SEQ ID NO:96.

7. The method of claim 1 , wherein the first polypeptide comprises the amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:97, SEQ ID NO:98, and SEQ ID NO:99 and the second polypeptide comprises the amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:96, SEQ ID NO:3, and SEQ ID NO:4.

8. The method of claim 1 , wherein the antigen-binding domain capable of specific binding to Fibroblast Activation Protein (FAP) is selected from the group consisting of an antibody fragment, a Fab molecule, a crossover Fab molecule, a single chain Fab molecule, a Fv molecule, a scFv molecule, a single domain antibody, an aVH, and a scaffold antigen-binding protein.

9. The method of claim 1 , wherein the TNF family ligand trimer-containing antigen-binding molecule comprises one antigen-binding domain capable of specific binding to Fibroblast Activation Protein (FAP).

10. The method of claim 1 , wherein the antigen-binding domain capable of specific binding to Fibroblast Activation Protein (FAP) is a Fab molecule.

11. The method of claim 1 , wherein the Fc domain is an IgG domain.

12. The method of claim 1 , wherein the Fc domain is an IgG1 Fc domain comprising amino acid substitutions at positions 234 and 235 (EU numbering) and/or 329 (EU numbering) of the IgG heavy chain.

13. The method of claim 1 , wherein the Fc domain is an IgG1 Fc domain or an IgG4 Fc domain.

14. The method of claim 1 , wherein the TNF family ligand trimer-containing antigen-binding molecule activates the NFκB signaling pathway.

15. The method of claim 1 , wherein the VH comprises (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:7 or SEQ ID NO:100, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:8 or SEQ ID NO:101, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:9 or SEQ ID NO:102, and the VL comprises (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:10 or SEQ ID NO:103, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:11 or SEQ ID NO:104, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:12 or SEQ ID NO:105.

16. The method of claim 1 , wherein the VH comprises the amino acid sequence of SEQ ID NO:16 and the VL comprises the amino acid sequence of SEQ ID NO:17 or wherein the VH comprises the amino acid sequence of SEQ ID NO:106 and the VL comprises the amino acid sequence of SEQ ID NO:107.

17. The method of claim 1 , the first polypeptide comprises a CH3 domain and the second polypeptide comprises a CH3 domain, wherein the first polypeptide comprises two ectodomains of the TNF family ligand, or fragments thereof, that are connected to each other and to the C-terminus of the CH3 domain by a peptide linker, and wherein the second polypeptide comprises only one ectodomain of the TNF family ligand, or fragment thereof, connected to the C-terminus of the CH3 domain of the second polypeptide by a peptide linker.

18. The method of claim 17 , wherein the TNF family ligand trimer-containing antigen-binding molecule comprises two antigen-binding domains capable of

Assignees

• Hoffmann La Roche

Inventors

• Amann Maria
• Bruenker Peter
• Claus Christina
• Ferrara Koller Claudia
• Grau-Richards Sandra
• Klein Christian
• Levitski Viktor
• Moessner Ekkehard
• Regula Joerg Thomas
• Umana Pablo

Classifications

• C07K2317/55

  Fab or Fab' · CPC title

• C07K16/2803

  against the immunoglobulin superfamily · CPC title

• C07K16/3007 Primary

  Carcino-embryonic Antigens · CPC title

• C07K16/40 Primary

  against enzymes · CPC title

• C07K14/70596

  Molecules with a "CD"-designation not provided for elsewhere · CPC title