Antibodies binding to fibroblast activation protein alpha and death receptor 4
Inventors
DE GOEIJ, Bart E.C.G. • Jongerius, Ilse • ANDRINGA, Grietje • Paauwe, Madelon • Plantinga, Theodorus Sjouke • Satijn, David • Laoukili, Jamila • Kranenburg, Onno Wouter • Overdijk, Marije
Assignees
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Publication Number
US-12351650-B2
Publication Date
2025-07-08
Expiration Date
Abstract
The present invention relates to a multispecific antibody comprising at least a FAPα binding region comprising a first heavy chain variable region and a first light chain variable region; and a DR4 binding region comprising a second heavy chain variable region and a second light chain variable region. The invention further provides pharmaceutical compositions comprising the antibodies and use of the antibodies for therapeutic and diagnostic procedures, in particular in cancer therapy.
Core Innovation
The invention relates to a bispecific antibody that comprises a fibroblast activation protein alpha (FAPα) binding region and a death receptor 4 (DR4) binding region. The FAPα binding region comprises a heavy chain variable (VH) region and a light chain variable (VL) region, and the DR4 binding region comprises a VH region and a VL region. The VH and VL regions are defined by complementarity determining regions CDR1, CDR2, and CDR3 within specific amino acid sequences set forth in SEQ ID NO: 13 and SEQ ID NO: 14 for FAPα, and SEQ ID NO: 15 and SEQ ID NO: 16 for DR4.
The antibody variants use Fc/CH3 interface residue changes to control heterodimer/bispecific Fab-arm exchange, using Eu numbering. Examples include heavy-chain substitutions L234F/L235E/D265A (FEA) or L234F/L235E/G236R (FER), together with CH3 mutations such as F405L and/or K409R. These design elements support heterodimeric interaction rather than homodimerization and regulate Fab-arm exchange behavior.
The described rationale links multispecific binding to conditional DR4 transactivation on DR4+/FAPα+ tumor microenvironment interactions and to trans-binding-dependent apoptosis, including mechanisms associated with caspase-8 and caspase-3 activity and downstream signaling involving FADD. The invention also describes pharmaceutical compositions, a companion diagnostic kit associated with the bispecific antibody, and cancer therapy, including solid cancers characterized by DR4 and FAPα expression.
The document further provides specified VH/VL CDR sequence sets and full-length sequences for anti-FAPα and anti-DR4 variants, including binding affinity metrics, comparative binding results, and cross-reactivity to FAPα/DR4 orthologs. Additional experimental examples described include binding to fibroblasts/CAFs and multiple tumor cell lines, PDO targeting, in vivo PDX models, and cell death-related mechanisms.
Claims Coverage
The independent claims present two inventive formulations of a bispecific antibody. Both independent claims define a FAPα binding region and a DR4 binding region using VH and VL regions whose CDR1, CDR2, and CDR3 sequences are fixed by SEQ ID numbers, and the broader content also describes Fc/CH3 interface residue changes for controlled Fab-arm exchange.
FAPα and DR4 bispecific binding regions with CDR-defined VH/VL sequences
A bispecific antibody comprising a FAPα binding region comprising a VH region and a VL region, and a DR4 binding region comprising a VH region and a VL region, wherein the FAPα VH region comprises the three complementarity determining regions CDR1, CDR2, and CDR3 present within the amino acid sequence set forth in SEQ ID NO: 13, the FAPα VL region comprises the three complementarity determining regions CDR1, CDR2, and CDR3 present within the amino acid sequence set forth in SEQ ID NO: 14, the DR4 VH region comprises CDR1, CDR2, and CDR3 present within the amino acid sequence set forth in SEQ ID NO: 15, and the DR4 VL region comprises CDR1, CDR2, and CDR3 present within the amino acid sequence set forth in SEQ ID NO: 16.
Controlled Fab-arm exchange through Fc/CH3 interface residue changes
Fc/CH3 interface residue changes using Eu numbering, including heavy-chain substitutions L234F/L235E/D265A (FEA) or L234F/L235E/G236R (FER), together with CH3 mutations such as F405L and/or K409R, are used to control heterodimer/bispecific Fab-arm exchange and support heterodimeric interaction rather than homodimerization.
Across the independent claims and related description, the core claim coverage is a FAPα/DR4 bispecific antibody whose binding regions are defined by CDR1/2/3 sequences in specified SEQ ID numbers, with controlled Fab-arm exchange supported by Fc/CH3 interface residue changes.
Stated Advantages
Reduced Fc-mediated effector function is described in the context of DR4 antagonist/effector-modulation concepts.
Reported comparative binding performance includes apparent affinity and EC50 reduction versus reference antibodies.
Enables conditional DR4 transactivation on DR4+/FAPα+ tumor microenvironment interactions.
Provides trans-binding-dependent apoptosis linked to caspase-8 and caspase-3 and associated signaling involving FADD.
Provides therapeutic compositions for treating solid cancers characterized by DR4 and FAPα expression.
Provides a companion diagnostic kit associated with the bispecific antibody.
Documented Applications
Treating cancer by administering an effective amount of the bispecific antibody to a subject in need.
Use in treating solid cancers with DR4 and FAPα expression, including CRC, pancreatic, gastric, breast, and NSCLC/TNBC as referenced in the provided content.
A diagnostic kit/companion diagnostic associated with the bispecific antibody.
Binding to fibroblasts/CAFs and binding to multiple tumor cell lines as part of the reported experimental evidence.
PDO targeting and in vivo PDX models are described as further experimental examples.
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