Antigen-binding molecules comprising a tumor necrosis factor (TNF) family ligand trimer

Patent metadata

Field	Value
Publication number	US-11267903-B2
Application number	US-201916522412-A
Country	US
Kind code	B2
Filing date	Jul 25, 2019
Priority date	Nov 14, 2014
Publication date	Mar 8, 2022
Grant date	Mar 8, 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 tumor necrosis factor (TNF) family ligand trimer-containing antigen-binding molecule comprising:
(a) at least one antigen-binding domain comprising an antibody light chain variable region (VL) and an antibody heavy chain variable region (VH) capable of specific binding to a target cell antigen, wherein the target cell antigen is CD19, and
(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 TNF family ligand is 4-1BBL.

2. The TNF family ligand trimer-containing antigen-binding molecule of claim 1 , further comprising:
(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.

3. The TNF family ligand trimer-containing antigen-binding molecule of claim 1 , wherein
(i) the first polypeptide comprises an antibody heavy chain constant 1 (CH1) domain or an antibody 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 an antibody 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 an antibody heavy chain variable region-light chain constant domain (VH-CL) or an antibody 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 TNF family ligand trimer-containing antigen-binding molecule of claim 1 , wherein the TNF family ligand costimulates human T-cell activation.

5. The TNF family ligand trimer-containing antigen-binding molecule of claim 1 , wherein the ectodomain of the TNF family ligand or fragment thereof comprises an 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.

6. The TNF family ligand trimer-containing antigen-binding molecule 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 TNF family ligand trimer-containing antigen-binding molecule of claim 1 , wherein the first polypeptide comprises an 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 an 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 TNF family ligand trimer-containing antigen-binding molecule of claim 1 , wherein the antigen-binding domain capable of specific binding to CD19 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 TNF family ligand trimer-containing antigen-binding molecule of claim 1 , wherein the molecule comprises one antigen-binding domain capable of specific binding to CD19.

10. The TNF family ligand trimer-containing antigen-binding molecule claim 1 , wherein the antigen-binding domain capable of specific binding to CD19 is a Fab molecule.

11. The TNF family ligand trimer-containing antigen-binding molecule of claim 2 , wherein the Fc domain is an IgG domain.

12. The TNF family ligand trimer-containing antigen-binding molecule of claim 2 , 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 TNF family ligand trimer-containing antigen-binding molecule of claim 1 , wherein 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.

14. The TNF family ligand trimer-containing antigen-binding molecule of claim 13 , comprising two antigen-binding domains capable of specific binding to CD19.

15. The TNF family ligand trimer-containing antigen-binding molecule of claim 1 , wherein the VH comprises (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO:195 or SEQ ID NO:252, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO:196 or SEQ ID NO:253, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO:197 or SEQ ID NO:254, and the VL comprises (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO:198 or SEQ ID NO:249, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO:199 or SEQ ID NO:250, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO:200 or SEQ ID NO:251.

16. The TNF family ligand trimer-containing antigen-binding molecule of claim 1 , wherein the VH comprises the amino acid sequence of SEQ ID NO:201 and the VL comprises the amino acid sequence of SEQ ID NO:202 or wherein the VH comprises the amino acid sequence of SEQ ID NO:357 and the VL comprises the amino acid sequence of SEQ ID NO:358.

17. The TNF family ligand trimer-containing antigen-binding molecule of claim 1 , wherein the ectodomain of the TNF family ligand or fragment thereof comprises the amino acid sequence of SEQ ID NO:1.

18. The TNF family ligand trimer-containing antigen-binding molecule of claim 2 , wherein the Fc domain is an IgG1 Fc domain or an IgG4 Fc domain.

19. The TNF family ligand trimer-containing antigen-binding molecule of claim 1 , wherein the antigen-binding molecule activate

Assignees

• Hoffmann La Roche
• Hofmann La Roche Inc

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

• C07K2317/33

  Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity · CPC title

• C07K14/70596

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