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CN-122029194-A - Novel TM4SF5 specific antibody

CN122029194ACN 122029194 ACN122029194 ACN 122029194ACN-122029194-A

Abstract

The present invention relates to an antibody or an antigen-binding fragment thereof which specifically binds to TM4SF5 and is thus useful for the prevention, treatment and diagnosis of diseases caused by overexpression of TM4SF 5.

Inventors

  • JIN SHIMEI
  • LI ZHENGYUAN
  • JIN YINMEI
  • GAO DONGJUN
  • YIN ZHENHUA

Assignees

  • 韩国生命工学研究院
  • 首尔大学校产学协力团

Dates

Publication Date
20260512
Application Date
20240712
Priority Date
20230714

Claims (13)

  1. 1. A TM4SF5 binding antibody or antigen binding fragment thereof comprising a CDR-comprising variable domain selected from the group consisting of: (i) A heavy chain variable domain (VH) comprising HCDR1 having the amino acid sequence of SEQ ID NO. 16, HCDR2 having the amino acid sequence of SEQ ID NO. 17 and HCDR3 having the amino acid sequence of SEQ ID NO. 18, and a light chain variable domain (VL) comprising LCDR1 having the amino acid sequence of SEQ ID NO. 11, LCDR2 having the amino acid sequence of SEQ ID NO. 12 and LCDR3 having the amino acid sequence of SEQ ID NO. 13; (ii) A heavy chain variable domain (VH) comprising HCDR1 having the amino acid sequence of SEQ ID NO. 24, HCDR2 having the amino acid sequence of SEQ ID NO. 25 and HCDR3 having the amino acid sequence of SEQ ID NO. 26, and a light chain variable domain (VL) comprising LCDR1 having the amino acid sequence of SEQ ID NO. 11, LCDR2 having the amino acid sequence of SEQ ID NO. 12 and LCDR3 having the amino acid sequence of SEQ ID NO. 13; (iii) A heavy chain variable domain (VH) comprising HCDR1 having the amino acid sequence of SEQ ID NO: 32, HCDR2 having the amino acid sequence of SEQ ID NO: 34 and HCDR3 having the amino acid sequence of SEQ ID NO: 34, and a light chain variable domain (VL) comprising LCDR1 having the amino acid sequence of SEQ ID NO:11, LCDR2 having the amino acid sequence of SEQ ID NO: 12 and LCDR3 having the amino acid sequence of SEQ ID NO: 13, and (Iv) A heavy chain variable domain (VH) comprising HCDR1 having the amino acid sequence of SEQ ID NO: 40, HCDR2 having the amino acid sequence of SEQ ID NO: 41 and HCDR3 having the amino acid sequence of SEQ ID NO: 42, and a light chain variable domain (VL) comprising LCDR1 having the amino acid sequence of SEQ ID NO: 48, LCDR2 having the amino acid sequence of SEQ ID NO: 49 and LCDR3 having the amino acid sequence of SEQ ID NO: 50, Wherein the CDR sequences are specified according to the Kabat numbering system.
  2. 2. The TM4SF5 binding antibody or antigen binding fragment thereof of claim 1, comprising a CDR-comprising variable domain selected from the group consisting of: (i) A heavy chain variable domain (VH) comprising HCDR1 having the amino acid sequence of SEQ ID NO. 16, HCDR2 having the amino acid sequence of SEQ ID NO. 17 and HCDR3 having the amino acid sequence of SEQ ID NO. 18, and a light chain variable domain (VL) comprising LCDR1 having the amino acid sequence of SEQ ID NO. 11, LCDR2 having the amino acid sequence of SEQ ID NO. 12 and LCDR3 having the amino acid sequence of SEQ ID NO. 13; (ii) A heavy chain variable domain (VH) comprising HCDR1 having the amino acid sequence of SEQ ID NO: 24, HCDR2 having the amino acid sequence of SEQ ID NO: 25 and HCDR3 having the amino acid sequence of SEQ ID NO: 26, and a light chain variable domain (VL) comprising LCDR1 having the amino acid sequence of SEQ ID NO: 11, LCDR2 having the amino acid sequence of SEQ ID NO: 12 and LCDR3 having the amino acid sequence of SEQ ID NO: 13, and (Iii) A heavy chain variable domain (VH) comprising HCDR1 having the amino acid sequence of SEQ ID NO. 32, HCDR2 having SEQ ID NO. 33 and HCDR3 having the amino acid sequence of SEQ ID NO. 34, and a light chain variable domain (VL) comprising LCDR1 having the amino acid sequence of SEQ ID NO. 11, LCDR2 having the amino acid sequence of SEQ ID NO. 12 and LCDR3 having the amino acid sequence of SEQ ID NO. 13, Wherein the CDR sequences are specified according to the Kabat numbering system.
  3. 3. The TM4SF5 binding antibody or antigen binding fragment thereof of claim 1, wherein the antibody or antigen binding fragment thereof comprises: i) A heavy chain variable domain having an amino acid sequence selected from the group consisting of SEQ ID NOS: 14, 22 and 30, and a light chain variable domain having an amino acid sequence of SEQ ID NO: 9, or Ii) a heavy chain variable domain having the amino acid sequence of SEQ ID NO. 38, and a light chain variable domain having the amino acid sequence of SEQ ID NO. 46.
  4. 4. The TM4SF5 binding antibody or antigen binding fragment thereof of claim 1, wherein the antibody is IgG or scFv-Fc.
  5. 5. A pharmaceutical composition for preventing or treating a TM4SF5 related disease, the pharmaceutical composition comprising the antibody or antigen binding fragment thereof of any one of claims 1-4.
  6. 6. The pharmaceutical composition of claim 5, wherein the TM4SF 5-related disease is selected from the group consisting of fatty liver, liver fibrosis, liver cancer, stomach cancer, colorectal cancer, rectal cancer, oral cancer, pharyngeal cancer, laryngeal cancer, lung cancer, colon cancer, breast cancer, cervical cancer, endometrial cancer, ovarian cancer, prostate cancer, testicular cancer, bladder cancer, kidney cancer, pancreatic cancer, bone cancer, connective tissue cancer, skin cancer, brain cancer, thyroid cancer, leukemia, hodgkin's disease, soft tissue sarcoma, lymphoma, and multiple myeloma blood cancer.
  7. 7. The pharmaceutical composition according to claim 5, wherein the TM4SF 5-related disease is selected from the group consisting of fatty liver, liver fibrosis, liver cancer, stomach cancer, colorectal cancer, colon cancer, prostate cancer, pancreatic cancer and soft tissue sarcoma.
  8. 8. The pharmaceutical composition of claim 5, wherein the pharmaceutical composition is administered in combination with a substance selected from the group consisting of 5-fluorouracil (5-FU), oxaliplatin, doxorubicin, sorafenib, and cetuximab.
  9. 9. A polynucleotide encoding the antibody or antigen-binding fragment thereof of any one of claims 1 to 4.
  10. 10. An expression vector comprising the polynucleotide of claim 9.
  11. 11. A transformant comprising the polynucleotide of claim 10 or an expression vector comprising the polynucleotide.
  12. 12. A composition for diagnosing a TM4SF5 related disease, the composition comprising the antibody or antigen-binding fragment thereof of any one of claims 1-4.
  13. 13. A method of providing information for diagnosing a TM4SF5 related disease, the method comprising detecting transmembrane 4L six family member 5 (TM 4SF 5) protein in a biological sample by an antigen-antibody reaction using the antibody or antigen binding fragment thereof of any one of claims 1-4.

Description

Novel TM4SF5 specific antibody Technical Field The invention relates to a TM4SF5 specific antibody and application thereof. Background Transmembrane 4 superfamily (transmembrane 4 superfamily,TM4SF) proteins are a group of hydrophobins with molecular weights of about 25kDa to 50kDa that include four transmembrane domains, two extracellular loops and two short cytoplasmic tail, also known as transmembrane tetra-protein (tetraspanin) or tetra-transmembrane protein (tetraspan). TM4SF proteins form complexes with cell adhesion molecules (e.g., integrins) on the cell membrane, thereby creating a large transmembrane tetra-protein rich microdomain (tetraspanin-enriched microdomain, TERM) and contributing to a variety of biological functions (e.g., cell adhesion, proliferation, differentiation, metabolic control, interactions with the immune system, and migration). TM4SF5 refers to transmembrane 4L six family member 5 (transmembrane 4L six family member 5) or four transmembrane L6 superfamily member 5 (Four-transmembrane L6 superfamily member 5), which are members of the transmembrane four protein family whose structure includes four insoluble protein domains that penetrate the cell membrane, two loops outside the cell, one loop and two tails in the cytoplasm. TM4SF5 is a homolog of tumor-associated antigen L6 (tumor-associated antigen L6, TM4SF 1), and mRNA of TM4SF1 is known to be highly overexpressed in pancreatic cancer, gastric cancer, liver cancer, colorectal cancer, soft tissue sarcoma, and the like. In addition, artificial expression of TM4SF5 protein in COS7 cells resulted in actin recombination (actin reorganization) and focal adhesion turnover (focal adhesion turnover), suggesting that TM4SF5 may be involved in cell migration through epithelial-mesenchymal transition (epithelial-MESENCHYMAL TRANSITION, EMT) (Lee s.a. et al, J CLIN INVEST 2008, 118 (4): 1354-66). In addition, since TM4SF5 protein has high amino acid sequence homology with L6 (a cancer-related gene), it has been suspected to be encoded by a cancer-related gene, and it has been reported to be related to the occurrence and progress of cancer. TM4SF5 promotes progression of the G1/S phase through intracellular expression of p27Kip1 and activity of RhoA GTPase, thereby participating in cell proliferation (Kim h.et al, biochim Biophys Acta 2010 1803 (8): 975-82), and crosstalk (cross-talk) in the signaling pathway between transforming growth factor- β1 (transforming growth factor- β1, tgf- β1) and Epidermal Growth Factor Receptor (EGFR) involved in EMT is known to induce expression of TM4SF5, thereby causing EMT (Kang m.et al., biochem J2012 443 (3): 691-700). Disclosure of Invention Technical problem As described above, with the increasing importance of TM4SF5 as a specific protein and anticancer target in new cancer diagnosis, a method of diagnosing cancer targeting TM4SF5 has been studied. In addition, studies for inhibiting the biological activity of TM4SF5 have also been conducted in various fields for cancer treatment targeting TM4SF 5. Specifically, studies have been conducted on compounds capable of inhibiting the activity of TM4SF5, for example, it has been reported that sulfonamide-substituted chalcone derivatives and sulfonate-substituted chalcone derivatives inhibit the biological activity of TM4SF5 (Korean patent No. 10-0934706). In addition to the compounds described above that inhibit the biological activity of TM4SF5, the importance of developing antibody therapeutics capable of specifically binding TM4SF5 is also growing. Accordingly, the present inventors have developed a humanized antibody capable of specifically binding to human TM4SF5 protein and have further improved it, thereby completing the present invention. Technical proposal It is an object of the present invention to provide an antibody or antigen-binding fragment thereof that binds TM4SF 5. It is another object of the present invention to provide a pharmaceutical composition for preventing or treating a TM4SF 5-related disease, which comprises the antibody or antigen-binding fragment thereof. It is a further object of the present invention to provide a composition for diagnosing a TM4SF5 related disease, comprising the antibody or antigen binding fragment thereof. It is a further object of the present invention to provide a polynucleotide encoding the antibody or antigen-binding fragment thereof and a transformant comprising the polynucleotide. Advantageous effects The antibodies of the invention are useful in the prevention, treatment and diagnosis of diseases in which TM4SF5 protein is overexpressed. Drawings Fig. 1 and 2 show the results of confirming physicochemical properties of huAb27-8 antibody, huAb27-9 antibody, huAb27-43 antibody and huAb27-69 antibody produced using the same vector and expression system as the chimeric antibody. (A of FIG. 1 and A of FIG. 2: SDS-PAGE; B of FIG. 1 and B of FIG. 2: SE-HPLC) FIG. 3 shows the results of protein analysis of h