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CN-122011189-A - Anti-CEACAM 5 antibody or antigen binding fragment thereof and application thereof

CN122011189ACN 122011189 ACN122011189 ACN 122011189ACN-122011189-A

Abstract

The invention relates to the technical field of biological medicines, in particular to an anti-CEACAM 5 antibody or an antigen binding fragment thereof and application thereof. The invention also relates to antibody-conjugated medicaments comprising an anti-CEACAM 5 antibody or antigen binding fragment thereof and their use in the treatment of tumors.

Inventors

  • QU AIDONG
  • ZHANG KUNMING
  • LIANG HONGYUAN
  • WU LINA
  • QIU JIANHUA

Assignees

  • 上海生物制品研究所有限责任公司

Dates

Publication Date
20260512
Application Date
20241108

Claims (20)

  1. 1. An anti-CEACAM 5 antibody or antigen-binding fragment thereof, wherein said anti-CEACAM 5 antibody or antigen-binding fragment thereof comprises a VHCDR1-3 of a heavy chain variable region and/or a VLCDR1-3 of a light chain variable region, wherein: The amino acid sequence of VHCDR1 comprises the amino acid sequence shown in SEQ ID NO 16, 22, 28, 34, 40 or 46; The amino acid sequence of VHCDR2 comprises the amino acid sequence shown in SEQ ID NO 17, 23, 29, 35, 41 or 47; the amino acid sequence of VHCDR3 comprises the amino acid sequence shown in SEQ ID NO 18, 30, 36 or 48; The amino acid sequence of VLCDR1 comprises the amino acid sequence shown in SEQ ID NO 19, 31, 37 or 49; The amino acid sequence of VLCDR2 comprises the amino acid sequence shown as SEQ ID NO. 20, 32, 38 or 50; The amino acid sequence of VLCDR3 comprises the amino acid sequence shown in SEQ ID NO. 21 or 39.
  2. 2. The anti-CEACAM 5 antibody or antigen-binding fragment thereof of claim 1, wherein the amino acid sequences of VHCDR1-3 and VLCDR1-3 comprise:
  3. 3. The anti-CEACAM 5 antibody or antigen-binding fragment thereof of claim 1, wherein the amino acid sequences of VHCDR1-3 and VLCDR1-3 comprise:
  4. 4. the anti-CEACAM 5 antibody or antigen-binding fragment thereof of any one of claims 1-3, wherein said anti-CEACAM 5 antibody or antigen-binding fragment thereof further comprises an FR region, preferably VHFR1-4 comprising a heavy chain variable region and/or VLFR1-4 comprising a light chain variable region; preferably, the VHFR1-4 or VLFR1-4, respectively, is independently derived from a human or non-human animal.
  5. 5. The anti-CEACAM 5 antibody or antigen-binding fragment thereof as claimed in any one of claims 1-4, wherein the heavy chain variable region comprises, in order from N-terminus to C-terminus, VHFR1, VHCDR1, VHFR2, VHCDR2, VHFR3, VHCDR3, VHFR4, and/or, The light chain variable region comprises VLFR1, VLCDR1, VLFR2, VLCDR2, VLFR3, VLCDR3 and VLFR4 from the N terminal to the C terminal in sequence.
  6. 6. The anti-CEACAM 5 antibody or antigen-binding fragment thereof of any one of claims 1-5, wherein the amino acid sequence of the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID No. 4 or 6; The amino acid sequence of the light chain variable region comprises the amino acid sequence shown in SEQ ID NO. 5 or 7.
  7. 7. The anti-CEACAM 5 antibody or antigen-binding fragment thereof of any one of claims 1-6, wherein the anti-CEACAM 5 antibody or antigen-binding fragment thereof comprises Fab, fd, fab ', fab ' -SH, fv, scFv, F (ab ') 2, or a diabody (dAb).
  8. 8. The anti-CEACAM 5 antibody or antigen-binding fragment thereof of any one of claims 1-7, wherein said anti-CEACAM 5 antibody or antigen-binding fragment thereof comprises a heavy chain constant region and/or a light chain constant region, preferably wherein said heavy chain constant region and/or light chain constant region is derived from a human or non-human animal.
  9. 9. The anti-CEACAM 5 antibody or antigen-binding fragment thereof of claim 8, wherein the heavy chain constant region is selected from the group consisting of a constant region of any one of IgG, igD, igA, igM or IgE heavy chain, preferably wherein the IgG heavy chain comprises an IgG1 heavy chain, an IgG2b heavy chain, an IgG3 heavy chain, or an IgG4 heavy chain; Preferably, the amino acid sequence of the heavy chain constant region comprises SEQ ID NO. 10.
  10. 10. The anti-CEACAM 5 antibody or antigen-binding fragment thereof of claim 8 or 9, wherein the light chain constant region is selected from the constant region of any one of a lambda-type light chain or a kappa-type light chain; preferably, the amino acid sequence of the light chain constant region comprises SEQ ID NO. 11 or 12.
  11. 11. A method of preparing an anti-CEACAM 5 antibody or antigen-binding fragment thereof according to any one of claims 1-10, comprising: 1) Obtaining a nucleic acid encoding a CEACAM5 antibody or antigen binding fragment thereof; 2) Transforming the nucleic acid obtained in step 1) into a host cell and then inducing its expression.
  12. 12. A nucleic acid comprising a nucleotide sequence encoding the anti-CEACAM 5 antibody or antigen-binding fragment thereof of any one of claims 1-10.
  13. 13. A carrier, characterized in that, the vector comprises the nucleic acid of claim 12.
  14. 14. A host cell comprising the nucleic acid of claim 12 and/or the vector of claim 13.
  15. 15. Use of an anti-CEACAM 5 antibody or antigen-binding fragment thereof of any one of claims 1-10, the nucleic acid of claim 12, the vector of claim 13, or the host cell of claim 14 in the preparation of an antibody-conjugated drug.
  16. 16. An antibody conjugated drug comprising the anti-CEACAM 5 antibody or antigen-binding fragment thereof of any one of claims 1-10 and a small molecule drug.
  17. 17. The antibody conjugated drug of claim 16, wherein the small molecule drug comprises a cytotoxin, such as hydroxycamptothecin and derivatives thereof, cyclophosphamide, nitrogen mustard, maytansine, methyl australistatin E, methotrexate, mitomycin C, cytarabine, paclitaxel, vinorelbine or docetaxel.
  18. 18. The antibody conjugated drug of claim 16 or 17, wherein the anti-CEACAM 5 antibody or antigen-binding fragment thereof is site-specifically conjugated to a small molecule drug to obtain the antibody conjugated drug.
  19. 19. A method of preparing an antibody conjugated drug according to any one of claims 16 to 18, comprising introducing an azide group onto an anti-CEACAM 5 antibody or antigen binding fragment thereof according to any one of claims 1 to 10, and reacting the antibody conjugated drug with DBCO.
  20. 20. Use of an anti-CEACAM 5 antibody or antigen-binding fragment thereof according to any one of claims 1-10, a nucleic acid according to claim 12, a vector according to claim 13, a host cell according to claim 14 or an antibody-conjugated drug according to any one of claims 16-18 for the preparation of a product for the diagnosis and/or treatment of a tumor.

Description

Anti-CEACAM 5 antibody or antigen binding fragment thereof and application thereof Technical Field The invention relates to the technical field of biological medicines, in particular to an anti-CEACAM 5 antibody or an antigen binding fragment thereof and application thereof. Background Carcinoembryonic antigen-related cell adhesion molecule 5 (Carcinoembryonic antigen-RELATED CELL adhesion molecule, ceacam 5) is expressed in normal tissues with limited expression, but is highly expressed on the surface of various epithelial tumor cells. CEACAM5, also known as CD66e, gene is located on human chromosome 19q13.2, consisting of 9 exons and 3 non-coding exons. CEACAM5 activates endothelial cells and tumor angiogenesis by participating in regulation of cell differentiation, apoptosis and cell polarity, and the highly fucosylation of 28N-glycosylation sites on the surface of CEACAM5 protein can directly promote epithelial-mesenchymal transition (EPITHELIAL-MESENCHYMAL TRANSITION, EMT) and dedifferentiation, and research proves that the interaction between CEACAM5 on the surface of tumor cells and CEACAM1 on the surface of NK cells can inhibit NK cell-mediated antitumor immune effects, thus CEACAM5 is one of the key driving factors in the development process of tumorigenesis. CEACAM5 was originally discovered during the 20 th century 70 in the course of research into human embryo development, and a significant increase in CEACAM5 protein expression levels in a variety of tumors was observed in later studies, while CEACAM5 protein was shed from the tumor cell surface to form soluble CEACAM5, making it an effective clinical biomarker and potential therapeutic target in melanoma, lung cancer, colorectal cancer, gastric cancer, and pancreatic cancer. Over-expressing CEACAM5 in more than 90% of tumor tissues in gastric cancer patients, and significantly higher than normal tissues in 98.8% of colorectal cancers (colorectal cancer, CRC), CEACAM5 is a biomarker for monitoring tumor recurrence in colorectal cancer patients, and elevated expression levels are generally regarded as signs of high tumor burden and poor prognosis. In CRC patients, 5-year survival correlated inversely with CEACAM5 tissue expression is a prognostic biomarker and monitoring biomarker currently used clinically to determine treatment regimens. The invention aims at developing a novel specific antibody targeting CEACAM5, and has important significance for diagnosis and treatment of tumors. Disclosure of Invention In the invention, in order to obtain the antibodies of the membrane-proximal end of the cell adhesion molecule 5 (Carcinoembryonic antigen-RELATED CELL adhesion molecule, CEACAM5) related to the specific targeting carcinoembryonic antigen by screening, different fusion proteins are adopted to immunize mice, and a plurality of fusion proteins are designed for screening. Finally, a plurality of mouse monoclonal antibodies which are specifically combined with the CEACAM5 near-membrane end domain are obtained, and after chimeric modification, the specific combination capacity and in-vitro cytotoxicity activity of the antibodies at the protein and cell level are evaluated, so that the antibodies are proved to have excellent endocytosis and are expected to be further developed into antibody coupled drugs (Antibody Drug Conjugates, ADC). The ADC is further prepared by site-specific coupling small molecule drugs, and the result shows that the ADC has obvious killing effect on tumor cells. The specific scheme is as follows: In a first aspect of the invention there is provided an anti-CEACAM 5 antibody or antigen-binding fragment thereof, said anti-CEACAM 5 antibody or antigen-binding fragment thereof comprising a VHCDR1-3 of a heavy chain variable region and/or a VLCDR1-3 of a light chain variable region. Wherein: The amino acid sequence of VHCDR1 comprises a sequence of at least 4 consecutive amino acids, preferably at least 4,5, 6, 7, 8, 9 or 10 consecutive amino acids in GYTFTNYGMN (SEQ ID NO: 55). Or the amino acid sequence of VHCDR1 comprises a sequence of at least 4 consecutive amino acids, preferably at least 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18 consecutive amino acids in GYIFTNYGMS (SEQ ID NO: 56). In a specific embodiment of the invention, the amino acid sequence of VHCDR1 comprises the amino acid sequence of SEQ ID NO 16, 22, 28, 34, 40 or 46 or comprises an amino acid sequence having 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more or 99% or comprises an amino acid sequence having one, two, three, four, five, six, seven, eight, nine or ten amino acids substituted, deleted or added in the amino acid sequence of SEQ ID NO 16, 22, 28, 34, 40 or 46. The amino acid sequence of VHCDR2 comprises at least 4 consecutive amino acids, preferably at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17 consecutive a