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EP-4735115-A1 - ANTIBODIES BINDING TO FIBROBLAST ACTIVATION PROTEIN ALPHA AND DEATH RECEPTOR 4

EP4735115A1EP 4735115 A1EP4735115 A1EP 4735115A1EP-4735115-A1

Abstract

The present invention relates to a multispecific antibody comprising at least a FAPa 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.

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

  • Genmab A/S

Dates

Publication Date
20260506
Application Date
20240626

Claims (20)

  1. 1. A multispecific antibody comprising at least (i) a FAPa binding region comprising a first heavy chain variable region and a first light chain variable region; and (ii) a DR4 binding region comprising a second heavy chain variable region and a second light chain variable region.
  2. 2. The multispecific antibody according to claim 1, wherein the FAPa binding region comprises a heavy chain variable region (VH) comprising the three complementarity determining regions, CDR1, CDR2, and CDR3, present within the amino acid sequence set forth in SEQ ID NO: 13.
  3. 3. The multispecific antibody according to any one of the preceding claims, wherein the FAPa binding region comprises a light chain variable region (VL) comprising the three complementarity determining regions, CDR1, CDR2, and CDR3, present within the amino acid sequence set forth in SEQ ID NO: 14.
  4. 4. The multispecific antibody according to any one of the preceding claims, wherein the FAPa binding region comprises a heavy chain variable region (VH) comprising the three complementarity determining regions, CDR1, CDR2, and CDR3, present within the amino acid sequence set forth in SEQ ID NO: 13, and a light chain variable region (VL) comprising the three complementarity determining regions, CDR1, CDR2, and CDR3, present within the amino acid sequence set forth in SEQ ID NO: 14.
  5. 5. The multispecific antibody according to any one of the preceding claims, wherein the FAPa binding region comprises a heavy chain variable region (VH) comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs. : 1, 2 and 3, respectively.
  6. 6. The multispecific antibody according to any one of the preceding claims, wherein the FAPa binding region comprises a light chain variable region (VL) comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs. : 4, 5 and 6, respectively.
  7. 7. The multispecific antibody according to any one of the preceding claims, wherein the FAPa binding region comprises a heavy chain variable region (VH) comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs. : 1, 2 and 3, respectively, and a light chain variable region (VL) comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NOs. : 4, 5 and 6, respectively.
  8. 8. The multispecific antibody according to any of the preceding claims, wherein the VH sequence of the FAPa binding region has at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO. : 13.
  9. 9. The multispecific antibody according to any of the preceding claims, wherein said VL sequence of the FAPa binding region has at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO. : 14.
  10. 10. The multispecific antibody according to any of the preceding claims, wherein the VH sequence of the FAPa binding region has at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO. : 13 and said VL sequence of the FAPa binding region has at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO. : 14.
  11. 11. The multispecific antibody according to any of the preceding claims, wherein the framework regions of the VH sequence of the FAPa binding region has at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% amino acid sequence identity to the framework regions of the amino acid sequence as set forth in SEQ ID NO. : 13.
  12. 12. The multispecific antibody according to any of the preceding claims, wherein the framework regions of the VL sequence of the FAPa binding region has at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% amino acid sequence identity to the framework regions of the amino acid sequence as set forth in SEQ ID NO. : 14.
  13. 13. The multispecific antibody according to any of the preceding claims, wherein the framework regions of the VH sequence of the FAPa binding region has at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% amino acid sequence identity to the framework regions of the amino acid sequence as set forth in SEQ ID NO. : 13 and said framework regions of the VL sequence of the FAPa binding region has at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% amino acid sequence identity to the framework regions of the amino acid sequence as set forth in SEQ ID NO. : 14.
  14. 14. The multispecific antibody according to any one of the preceding claims, wherein said VH sequence of the FAPa binding region deviate from SEQ ID NO. : 13 by at the most 10 substitutions, such as at the most 9 substitutions, like at the most 8 substitutions, such as at the most 7 substitutions, like at the most 6 substitutions, such as at the most 5 substitutions, like at the most 4 substitutions, such as at the most 3 substitutions, like at the most 2 substitutions, such as at the most 1 substitution.
  15. 15. The multispecific antibody according to any one of the preceding claims, wherein said VL sequence of the FAPa binding region deviate from SEQ ID NO. : 14 by at the most 10 substitutions, such as at the most 9 substitution, like at the most 8 substitutions, such as at the most 7 substitutions, like at the most 6 substitutions, such as at the most 5 substitutions, like at the most 4 substitutions, such as at the most 3 substitutions, like at the most 2 substitutions, such as at the most 1 substitution.
  16. 16. The multispecific antibody according to any one of the preceding claims, wherein the VH and VL sequences of the FAPa binding region only deviate in the framework regions.
  17. 17. The multispecific antibody according to any of the preceding claims, wherein the VH sequence of the FAPa binding region comprises, consists essentially of or consists of a VH sequence as set forth in SEQ ID NO. : 13.
  18. 18. The multispecific antibody according to any of the preceding claims, wherein the VL sequence of the FAPa binding region comprises, consists essentially of or consists of a VL sequence as set forth in SEQ ID NO. : 14.
  19. 19. The multispecific antibody according to any of the preceding claims, wherein the VH and VL sequences of the FAPa binding region comprise, consist essentially of or consist of a VH sequence as set forth in SEQ ID NO. : 13 and a VL sequence as set forth in SEQ ID NO. : 14.
  20. 20. The multispecific antibody according to any one of the preceding claims, wherein FAPa is human FAPa such as the mature polypeptide of SEQ ID NO: 33 or a soluble FAPa of SEQ ID NO: 34; FAPa is mouse FAPa such as the mature polypeptide of SEQ ID NO: 35; FAPa is rat FAPa such as the mature polypeptide of SEQ ID NO: 36; FAPa is dog FAPa such as the mature polypeptide of SEQ ID NO: 37; FAPa is pig FAPa such as the mature polypeptide of SEQ ID NO: 38; or cynomolgus monkey FAPa, such as the mature polypeptide of SEQ ID NO: 39.

Description

ANTIBODIES BINDING TO FIBROBLAST ACTIVATION PROTEIN ALPHA AND DEATH RECEPTOR 4 Field of Invention The present invention relates to antibodies binding to Fibroblast Activation Protein Alpha (FAPa) and Death Receptor 4 (DR4). The invention further provides pharmaceutical compositions comprising the antibodies and use of the antibodies for therapeutic and diagnostic procedures, such as treatment of cancer. Background Therapeutic monoclonal antibodies are a promising type of immunotherapy due to their specific features which include target specificity, immune modulation, and generally low toxicity. Monoclonal antibodies are made of two distinct functional units: the antigen binding fragment Fab that binds to the targeted antigen, and the constant fragment (Fc) which mediates antibody-dependent immune effector functions. The main Fc domain- mediated mechanisms of action are complement dependent cytotoxicity (CDC) and binding to Fc-gamma receptors (specific for IgG) on various immune cells resulting in e.g. antibody-dependent cell-mediated cytotoxicity (ADCC). Additionally, Fab binding to the target can result in signaling perturbation. The knowledge of antibody-based therapeutic strategies has advanced dramatically over recent years, delivering breakthroughs on target biology, mechanisms of action but also on antibody formats and developments. This has led to antibody-based therapies aimed at enhancing clinical effectiveness, as well as improving the target specificity (and therefore safety). Apoptosis is a form of programmed cell death. Tumor necrosis factor (TNF)- related apoptosis-inducing ligand (TRAIL) can trigger apoptosis via binding to its agonist receptors that contain intracellular death domains (DD). DR4, also known as Tumor Necrosis Factor Receptor Superfamily member 10A (TNFRSF10A), TRAIL receptor 1 (TRAIL-R1) and CD261, is a cell surface receptor of the TNF receptor superfamily that binds TRAIL and mediates apoptosis. DR4 shares 60% homology with death receptor 5 (DR5), the other known TRAIL receptor capable of inducing apoptosis. DR4 is a singlepass type I membrane protein with at least three extracellular cysteine-rich domains (CRDs), a transmembrane domain (TM) and a cytoplasmic DD. TRAIL binding leads to DR4 activation via receptor trimerization, resulting in clustering of the DD, followed by recruitment of the Fas-associated death domain (FADD) adaptor protein. Next, FADD recruits caspase-8 and -10 to form the death-inducing signaling complex (DISC). Active caspase-8 and -10 are then released in the cytosol, where they activate downstream effector caspases such as caspase-3. Activation of effector caspases culminates with apoptotic cell death. FAPa is a type-II transmembrane (homodimer) serine protease, that is overexpressed in pathological conditions including fibrosis, arthritis, and cancer. FAPa is primarily expressed by activated stromal fibroblasts such as CAFs. FAPa can also be shed from the cell membrane forming soluble FAPa. FAPa is a member of the prolyl peptidase family and shares 70% amino acid sequence homology with the well-described dipeptidyl peptidase 4 (DPP4). Both peptidases contain dipeptidyl peptidase enzymatic activity, whilst endopeptidase activity is FAPa-specific and targets substrates including denatured collagen and o-2 anti-plasmin. While for most cancers elevated FAPa expression is associated with a worse outcome, the underlying biological mechanisms remain poorly understood. Several first-generation DR agonist antibodies have been tested in the clinic showing limited antitumor efficacy, likely due to their inability to induce efficient receptor clustering which is essential to trigger apoptosis (Dubuisson and Micheau, Antibodies (Basel) 6(4), 2017). One such example is mapatumumab (HGS-ETR1), a DR4-specific agonist monoclonal antibody which showed limited clinical activity as explored in multiple phase 1/2 trials (Snajdauf et al., Front Mol Biosci 8: 628332, 2021). Next-generation agents, such as the TRAIL-R agonist Fc-fusion protein Eftozanermin alfa (ABBV-621), showed encouraging clinical activity but also induced side effects such as liver toxicity (Papadopoulos et al, Cancer Chemother Pharmacol 75(5):887-895, 2015; LoRusso et al., Invest New Drugs 40(4): 762-772, 2022; Di Cristofano et al., Biochem Soc Trans 51(1): 57-70, 2023). The dual targeting of DR and FAPa has been explored with RG7386, an optimized tetravalent bispecific antibody targeting FAPa and DR5 (US9926379B2). However, clinical development for solid malignancies was stopped in 2018. Thus, there is a clear unmet need for development of novel therapeutics with improved safety and efficacy. Summary of Invention It is an object of the present invention to provide multispecific antibodies with improved safety and efficacy. The multispecific antibodies according to the present invention comprise a DR4 binding region and a FAPa binding region. Thus, the proposed mechanism of action for the multispecifi