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EP-4739716-A1 - EGFR/C-MET BISPECIFIC BINDING PROTEIN AND USE THEREOF

EP4739716A1EP 4739716 A1EP4739716 A1EP 4739716A1EP-4739716-A1

Abstract

Related to the field of disease treatment, and provided are bispecific antibodies against EGFR and c-MET, nucleic acid molecules encoding them, and methods for preparing them. Said anti-EGFR and c-MET bispecific antibody has high specificity and high affinity for EGFR and c-MET. Also provided is the use of said bispecific antibody in the treatment and diagnosis of diseases.

Inventors

  • ZHENG, YONG
  • SHAN, WEI
  • LI, Senwu
  • SONG, Hongmei
  • TAN, Miao
  • TAN, Xiangyang
  • GE, Junyou
  • LEI, Dongmei
  • ZHOU, Yuehua
  • DI, Yanshu
  • YI, Shuting
  • LONG, Hu
  • LUO, Shuntao
  • YUAN, Xiaoxi
  • LI, Mingxiong

Assignees

  • Sichuan Kelun-Biotech Biopharmaceutical Co., Ltd.

Dates

Publication Date
20260513
Application Date
20240705

Claims (20)

  1. A bispecific antibody, comprising a first antigen-binding domain specifically binding to c-MET and a second antigen-binding domain specifically binding to EGFR; wherein the first antigen-binding domain comprises a first light chain variable region (VL) and a first heavy chain variable region (VH) , and the first VL and the first VH collectively form a domain capable of specifically binding to c-MET; and the second antigen-binding domain comprises a second VL and a second VH, and the second VL and the second VH together form a domain capable of specifically binding to EGFR.
  2. The bispecific antibody of claim 1, wherein the first antigen-binding domain and the second antigen-binding domain are each independently selected from scFv, Fab, and scFab.
  3. The bispecific antibody according to claim 1 or 2, further comprising an Fc domain, wherein the Fc domain comprises a first Fc domain monomer and a second Fc domain monomer, and the first and second Fc domain monomers comprise one or more modifications that promote heterodimerization of the Fc domain monomers.
  4. The bispecific antibody of claim 3, wherein the Fc domain comprises a first Fc domain monomer comprising modifications that form a knob structure and a second Fc domain monomer comprising modifications that form a hole structure, wherein the hole structure can be paired with the knob structure to form a heterodimeric Fc domain.
  5. The bispecific antibody of claim 4, wherein the first Fc domain monomer comprises the amino acid sequence as shown in SEQ ID NO: 49 or 51, and the second Fc domain monomer comprises the amino acid sequence as shown in SEQ ID NO: 50 or 52.
  6. The bispecific antibody of any one of claims 1-5, wherein the first antigen-binding domain and the second antigen-binding domain are each linked to one of the first and second Fc domain monomers of the Fc domain.
  7. The bispecific antibody of claim 6, wherein the first antigen-binding domain is linked to the first Fc domain monomer, and the second antigen-binding domain is linked to the second Fc domain monomer; or the first antigen-binding domain is linked to the second Fc domain monomer, and the second antigen-binding domain is linked to the first Fc domain monomer.
  8. The bispecific antibody of any one of claims 1-7, wherein the first VL comprises LCDR1, LCDR2, and LCDR3 amino acid sequences of a VL amino acid sequence set forth in SEQ ID NO: 17 or 59; and/or the first VH comprises HCDR1, HCDR2, and HCDR3 amino acid sequences of a VH amino acid sequence set forth in SEQ ID NO: 18 or 60.
  9. The bispecific antibody of any one of claims 1-8, wherein: the first VL comprises: (i) LCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 34, LCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 36, and LCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 38; or (ii) LCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 35, LCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 37, and LCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 38; and/or the first VH comprises: (i) HCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 39, HCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 43, and HCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 47; (ii) HCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 40, HCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 44, and HCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 47; (iii) HCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 42, HCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 46, and HCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 47; or (iv) HCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 41, HCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 45, and HCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 48.
  10. The bispecific antibody of any one of claims 1-9, wherein: the first VL comprises the amino acid sequence as shown in SEQ ID NO: 17 or 59, and/or the first VH comprises the amino acid sequence as shown in SEQ ID NO: 18 or 60; or the first VL comprises the amino acid sequence as shown in SEQ ID NO: 17, the first VH comprises the amino acid sequence as shown in SEQ ID NO: 18; or the first VL comprises the amino acid sequence as shown in SEQ ID NO: 59, the first VH comprises the amino acid sequence as shown in SEQ ID NO: 60.
  11. The bispecific antibody of any one of claims 1-10, wherein the second VL comprises the LCDR1, LCDR2, and LCDR3 amino acid sequences of a VL amino acid sequence set forth in SEQ ID NO: 15; and/or the VH comprises the HCDR1, HCDR2, and HCDR3 amino acid sequences of a VH amino acid sequence set forth in SEQ ID NO: 16.
  12. The bispecific antibody of any one of claims 1-11, wherein: the second VL comprises: (i) LCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 19, LCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 21, and LCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 23; or (ii) LCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 20, LCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 22, and LCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 23; and/or the second VH comprises: (i) HCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 24, HCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 28, and HCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 32; (ii) HCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 29, and HCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 32; (iii) HCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 27, HCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 31, and HCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 32; or (iv) HCDR1 comprising the amino acid sequence as shown in SEQ ID NO: 26, HCDR2 comprising the amino acid sequence as shown in SEQ ID NO: 30, and HCDR3 comprising the amino acid sequence as shown in SEQ ID NO: 33.
  13. The bispecific antibody of any one of claims 1-12, wherein the second VL comprises the amino acid sequence as shown in SEQ ID NO: 15, and/or the second VH comprises the amino acid sequence as shown in SEQ ID NO: 16.
  14. The bispecific antibody of any one of claims 1-13, wherein the first antigen-binding domain is a Fab, and the second antigen-binding domain is an scFv.
  15. The bispecific antibody of claim 14, wherein the bispecific antibody comprises a peptide chain I-A, a peptide chain I-B and a peptide chain I-C; wherein the peptide chain I-A comprises the first VL and a light chain constant region; the peptide chain I-B comprises: the VH, a heavy chain CH1 region and the first Fc domain monomer (or the second Fc domain monomer) ; the peptide chain I-C comprises: the second VL, the second VH and the second Fc domain monomer (or the first Fc domain monomer) .
  16. The bispecific antibody of claim 15, wherein the peptide chain I-A comprises from N-terminus to C-terminus the first VL and the light chain constant region; the peptide chain I-B comprises from N-terminus to C-terminus the first VH, the heavy chain CH1 region and the first Fc domain monomer (or the second Fc domain monomer) ; and/or the peptide chain I-C comprises from N-terminus to C-terminus (i) the second VL, the second VH and the second Fc domain monomer (or the first Fc domain monomer) , or (ii) the second VH, the second VL and the second Fc domain monomer (or the first Fc domain monomer) .
  17. The bispecific antibody of claim 15 or 16, wherein the adjacent domains of the peptide chain I-A are connected optionally with or without a linker, and the adjacent domains of the peptide chain I-B are connected optionally with or without a linker, and/or the adjacent domains of the peptide chain I-C are connected optionally with or without a linker.
  18. The bispecific antibody of claim 17, wherein the linkers are each independently the same peptide linker or different peptide linkers (e.g., rigid peptide linkers or flexible peptide linkers) ; or the peptide linkers are each independently selected from the group consisting of peptide linkers comprising one or more glycines (G) and/or serines (S) , for example, having the structure shown as (GGGGS) n , wherein n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 (SEQ ID NOs: 55-58 or 63-68) ; or the peptide linkers each independently comprise the amino acid sequence shown in SEQ ID NOs: 55-58, 61, or 63-68.
  19. The bispecific antibody of any one of claims 15-18, wherein the light chain constant region comprises the amino acid sequence as shown in SEQ ID NO: 53, and/or the heavy chain CH1 region comprises the amino acid sequence as shown in SEQ ID NO: 54.
  20. The bispecific antibody of any one of claims 15-19, wherein the peptide chain I-A comprises the amino acid sequence as shown in SEQ ID NO: 1, the peptide chain I-B comprises the amino acid sequence as shown in SEQ ID NO: 2 or 9, and/or the peptide chain I-C comprises the amino acid sequence as shown in SEQ ID NO: ID NO: 3 or 10.

Description

EGFR/C-MET BISPECIFIC BINDING PROTEIN AND USE THEREOF FIELD OF THE INVENTION The present disclosure relates to therapeutic bispecific antibodies, and more specifically, to bispecific antibodies against EGFR and c-MET and methods of use thereof. DESCRIPTION OF RELATED ART EGFR (epidermal growth factor receptor, abbreviated as EGFR, ErbB-1 or HER1) is a member of the epidermal growth factor receptor (HER) family. This family includes HER1 (erbB1, EGFR) , HER2 (erbB2, NEU) , HER3 (erbB3) and HER4 (erbB4) . The HER family plays an important regulatory role in cellular physiological processes. EGFR is a type I transmembrane glycoprotein composed of 1210 amino acids, with a molecular weight of about 170KD. Its main structure includes a ligand-binding extracellular domain, a hydrophobic transmembrane region, and an intracellular region comprising tyrosine kinase. EGFR is highly expressed in various tumors such as colorectal cancer, head and neck cancer, and non-small cell lung cancer, and it is also expressed in normal epithelial tissues such as skin and lungs. Highly expressed EGFR in tumor tissue undergoes homologous or heterologous dimerization through the binding of ligands such as EGF or TGFα, and the dimerization leads to the activation of tyrosine kinases and protein phosphorylation in tumor cells, mediating gene transcription and activation of various cell signaling pathways in cell cycle progression, which plays an important role in promoting the survival, proliferation and migration of tumor cells and tumor angiogenesis. Tyrosine protein kinase Met (c-MET) , also known as hepatocyte growth factor receptor (HGFR) , is a heterodimeric transmembrane tyrosine kinase receptor encoded by the Met proto-oncogene. c-MET is a type I transmembrane glycoprotein composed of 1390 amino acids, with a molecular weight of about 190KD, and comprises two polypeptide chains connected by disulfide bonds through cleavage, namely the α chain (50 kDa) and the β chain (140 kDa) . MET protein can be divided into SEMA domain (Semaphorin domain, SEMA) , PSI domain (Plexin-semaphorin-integrin domain, PSI) , four immunoglobulin-like repeat domains (Immunoglobulin-plexins-transcription domain, IPT) , a transmembrane region, a juxtamembrane region (JM) , a tyrosine kinase domain (TK) and a carboxy-terminal tail region (CT) . The natural ligand for the c-MET receptor is hepatocyte growth factor (HGF) , an inactive protein that is converted to its active form by proteolytic cleavage. Aberrant regulation of c-MET has been reported in a variety of cancers, including colorectal cancer, non-small cell lung cancer, gastric cancer, and breast  cancer. High activation of c-MET and its downstream signaling pathways have been shown to trigger hyperproliferation, tumor invasion, and angiogenesis, and be associated with poor survival. Anti-EGFR macromolecular antibody drugs, such as cetuximab, panitumumab, nimotuzumab, and necitumumab have been approved for marketing, targeting colorectal cancer, non-small cell lung cancer, etc. EGFR-tyrosine kinase inhibitors (TKIs) , such as osimertinib, alimertinib, and avitinib, have become the standard treatment for advanced cancer patients positive for EGFR mutation. c-MET-TKIs have also achieved good efficacy in non-small cell lung cancer. The EGFR and c-MET signaling pathways have been clinically verified respectively. Nearly 60%of tumors resistant to EGFR-TKIs had increased c-MET expression, amplified c-MET, or increased c-MET ligand HGF (Turke et al., Cancer Cell, 17: 77-88, 2010) , suggesting that c-MET signaling compensates for EGFR signaling, in the tumor cell. Both EGFR and c-MET signal through the same survival and anti-apoptotic pathways (ERK and AKT) . The bispecific antibody targeting EGFR and c-MET at the same time is expected to simultaneously inhibit the signaling pathways of EGFR and c-MET, avoiding the occurrence and development of tumors caused by the activation of c-MET signaling pathway after EGFR inhibition, thereby improving the overall efficacy and safety. Anti-c-MET and anti-EGFR antibodies are known in the art, for example, Telisotuzumab vedotin (Teliso-v or ABBV399) , which is disclosed in U. S. Patent No. 10603389, is anti-c-MET antibody ABT-700 conjugated to vc-MMAE (vedotin) , and MRG003 (ADC-3 or MYK-3) , which is disclosed in WO2023040941, is anti-EGFR antibody BA03 conjugated to vedotin. Several EGFR/c-Met bispecific antibodies have been disclosed in the art. Bispecific antibody BSAB01 disclosed in WO2010115551 comprises the EGFR binding VH/VL pair (Fab) of cetuximab and the c-Met binding VH/VL pair (Fab) of onartuzumab, which is currently in Phase III trials. Amivantamab, a bispecific antibody marketed under the tradenameis disclosed in U.S. Patent No. 9593164. Amivantamab comprises an Anti-EGFR Fab comprising the VH and VL of Zalutumumab (See, U.S. Patent No. 7247301 and U.S. Patent No. 7595378) and an anti-cMET Fab comprising the VH and VL of antibody 069 disclosed