Antigen binding molecules comprising a TNF family ligand trimer and a Tenascin binding moiety

Patent metadata

Field	Value
Publication number	US-11149083-B2
Application number	US-201816186443-A
Country	US
Kind code	B2
Filing date	Nov 9, 2018
Priority date	May 11, 2016
Publication date	Oct 19, 2021
Grant date	Oct 19, 2021

Abstract

The invention relates to novel TNF family ligand trimer-containing antigen binding molecules comprising (a) at least one antigen binding moiety capable of specific binding to Tenascin-C (TnC) and (b) a first and a second polypeptide that are linked to each other by a disulfide bond, wherein the antigen binding molecules are characterized in that the first polypeptide comprises two ectodomains of a TNF ligand family member or two 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. The invention further relates to methods of producing these molecules and to methods of using the same.

First claim

The invention claimed is:

1. A TNF (tumor necrosis factor) family ligand trimer-containing antigen binding molecule comprising
(a) at least one antigen binding moiety capable of specific binding to Tenascin-C(TnC) and
(b) a first and a second polypeptide that are linked to each other by a disulfide bond,
wherein the antigen binding molecule is characterized in that the first polypeptide comprises two ectodomains of a TNF ligand family member or a fragment 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.

2. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , further comprising (c) an Fc domain composed of a first and a second subunit capable of stable association.

3. The TNF family ligand trimer-containing antigen binding molecule of claim 1 ,
wherein the antigen binding molecule is further characterized in that
(i) the first polypeptide contains a CH1 or CL domain and the second polypeptide contains a CL or CH1 domain, respectively, wherein the second polypeptide is linked to the first polypeptide by a disulfide bond between the CH1 and CL domain, or
(ii) the first polypeptide contains a CH3 domain and the second polypeptide contains a CH3 domain, respectively, or
(iii) the first polypeptide contains a VH-CL or a VL-CH1 domain and the second polypeptide contains a VL-CH1 domain or a VH-CL domain, respectively, wherein the second polypeptide is linked to the first polypeptide by a disulfide bond between the CH1 and CL domain.

4. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the TNF ligand family member costimulates human T-cell activation.

5. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the TNF ligand family member is selected from 4-1BBL and OX40L.

6. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the TNF ligand family member is 4-1BBL.

7. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the moiety capable of specific binding to TnC 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.

8. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the molecule comprises one or two moieties capable of specific binding to TnC.

9. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the antigen binding moiety capable of specific binding to TnC is a Fab molecule capable of specific binding to TnC.

10. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the antigen binding moiety capable of specific binding to TnC comprises a VH domain comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 67, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 68, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 69, and a VL domain comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 55, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 56, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 57.

11. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the antigen binding moiety capable of specific binding to TnC comprises a VH domain comprising (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 70, (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 71, and (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 72, and a VL domain comprising (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 58, (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 59, and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 60.

12. The TNF family ligand trimer-containing antigen binding molecule of claim 2 , wherein the Fc domain is an IgG1 Fc domain comprising the amino acid substitutions at positions 234 and 235 (EU numbering) and/or 329 (EU numbering).

13. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the antigen binding molecule comprises a first heavy chain and a first light chain, both comprising a Fab molecule capable of specific binding to TnC,
a first peptide comprising two ectodomains of a TNF ligand family member or fragments thereof connected to each other by a first peptide linker fused at its C-terminus by a second peptide linker to a second heavy or light chain, and a second peptide comprising one ectodomain of said TNF ligand family member fused at its C-terminus by a third peptide linker to a second light or heavy chain, respectively.

14. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the first peptide comprising two ectodomains of a TNF ligand family member or fragments thereof connected to each other by a first peptide linker is fused at its C-terminus by a second peptide linker to a CH1 domain that is part of a heavy chain, and the second peptide comprising one ectodomain of said TNF ligand family member or a fragment thereof is fused at its C-terminus by a third peptide linker to a CL domain that is part of a light chain.

15. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the first peptide comprising two ectodomains of a TNF ligand family member or fragments thereof connected to each other by a first peptide linker is fused at its C-terminus by a second peptide linker to a CL domain that is part of a heavy chain, and the second peptide comprising one ectodomain of said TNF ligand family member or a fragment thereof is fused at its C-terminus by a third peptide linker to a CH1 domain that is part of a light chain.

16. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the first peptide comprising two ectodomains of a TNF ligand family member or fragments thereof connected to each other by a first peptide linker is fused at its C-terminus by a second peptide linker to a VH domain that is part of a heavy chain, and the second peptide comprising one ectodomain of said TNF ligand family member or a fragment thereof is fused at its C-terminus by a third peptide linker to a VL domain that is part of a light chain.

17. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the antigen binding moiety binds to human TnC, and cross-reacts to mouse and cynomolgus TnC.

18. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the antigen binding moiety binds to at least one of human, mouse and cynomolgus TnC with a K D range of about 1 μM to about 0.001 nM.

19. The TNF family ligand trimer-containing antigen binding molecule of claim 17 , wherein the antigen binding moiety binds to human TnC with a K D range of about 1 μM to about 0.001 nM, and cross-reacts to mouse and cynomolgus TnC with a binding affinity within the K D range of a factor of 20 relative to the K D for binding human TnC.

20. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , for use as a medicament.

21. The TNF family ligand trimer-containing antigen binding molecule of claim 1 , for use in the treatment of cancer.

22. The TNF family ligand trimer-containing

Assignees

• Hoffmann—La Roche Inc
• Hoffmann La Roche

Inventors

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

Classifications

• A61K47/6813 Primary

  the drug being a peptidic cytokine, e.g. an interleukin or interferon · CPC title

• C07K14/7151

  for tumor necrosis factor [TNF], for lymphotoxin [LT] · CPC title

• C07K16/241

  Tumor Necrosis Factors · CPC title

• A61K47/6845

  the antibody targeting a cytokine, e.g. growth factors, VEGF, TNF, a lymphokine or an interferon · CPC title

• C07K2319/30

  Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto · CPC title