Antigen binding molecules comprising a tnf family ligand trimer

Patent metadata

Field	Value
Publication number	US-2016200833-A1
Application number	US-201615067024-A
Country	US
Kind code	A1
Filing date	Mar 10, 2016
Priority date	Nov 14, 2014
Publication date	Jul 14, 2016
Grant date	—

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 TNF family ligand trimer-containing antigen binding molecule 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, 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 , 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, wherein the antigen binding molecule is 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, and wherein the first polypeptide comprises two ectodomains of a TNF ligand family member or fragments thereof that are connected to each other and to the CH1 or CL domain by a peptide linker and wherein the second polypeptide comprises one ectodomain of said TNF ligand family member or a fragment thereof connected via a peptide linker to the CL or CH1 domain of said polypeptide, or
(ii) the first polypeptide contains a CH3 domain and the second polypeptide contains a CH3 domain, respectively, and wherein the first polypeptide comprises two ectodomains of a TNF ligand family member 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 said TNF ligand family member or a fragment thereof connected via a peptide linker to C-terminus of the CH3 domain of said polypeptide, 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, and wherein the first polypeptide comprises two ectodomains of a TNF ligand family member or fragments thereof that are connected to each other and to to VH or VL by a peptide linker and wherein the second polypeptide comprises one ectodomain of said TNF ligand family member or a fragment thereof connected via a peptide linker to VL or VH of said polypeptide.

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 ectodomain of a TNF ligand family member 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, particularly the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:96.

8 . The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the ectodomain of a TNF ligand family member comprises the amino acid sequence of SEQ ID NO:96.

9 . The TNF family ligand trimer-containing antigen binding molecule of claim 1 , 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, wherein the antigen binding molecule is characterized in that 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 in that 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.

10 . The TNF family ligand trimer-containing antigen binding molecule of claim 1 , comprising
(a) at least one moiety capable of specific binding to a target cell antigen, and (b) a first polypeptide containing a CH1 or CL domain and a second polypeptide containing 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, and wherein the antigen binding molecule is characterized in that the first polypeptide comprises two ectodomains of a TNF ligand family member or fragments thereof that are connected to each other and to the CH1 or CL domain by a peptide linker and in that the second polypeptide comprises only one ectodomain of said TNF ligand family member or a fragment thereof connected via a peptide linker to the CL or CH1 domain of said polypeptide.

11 . The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the moiety capable of specific binding to a target cell antigen 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.

12 . The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the molecule comprises one moiety capable of specific binding to a target cell antigen.

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

14 . The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the target cell antigen is selected from the group consisting of Fibroblast Activation Protein (FAP), Melanoma-associated Chondroitin Sulfate Proteoglycan (MCSP), Epidermal Growth Factor Receptor (EGFR), Carcinoembryonic Antigen (CEA), CD19, CD20 and CD33.

15 . The TNF family ligand trimer-containing antigen binding molecule of claim 1 , wherein the target cell antigen is Fibroblast Activation Protein (FAP).

16 . The TNF family ligand trimer-containing antigen binding molecule of claim 15 , wherein the moiety capable of specific binding to FAP comprises a VH domain comprising (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 a VL domain comprising (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.

17 . The TNF family ligand trimer-containing antigen binding molecule of claim 15 , wherein the moiety capable of specific binding to FAP comprises a variable heavy chain compris

Assignees

• Hoffmann La Roche

Inventors

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

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title

• C07K16/2803

  against the immunoglobulin superfamily · CPC title

• C07K16/3007 Primary

  Carcino-embryonic Antigens · CPC title

• C07K14/70575

  NGF/TNF-superfamily, e.g. CD70, CD95L, CD153, CD154 (NGF C07K14/48, TNF C07K14/525) · CPC title