WO2021091359A1 - 조절 t 세포 표면 항원의 에피토프 및 이에 특이적으로 결합하는 항체 - Google Patents

조절 t 세포 표면 항원의 에피토프 및 이에 특이적으로 결합하는 항체 Download PDF

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WO2021091359A1
WO2021091359A1 PCT/KR2020/015656 KR2020015656W WO2021091359A1 WO 2021091359 A1 WO2021091359 A1 WO 2021091359A1 KR 2020015656 W KR2020015656 W KR 2020015656W WO 2021091359 A1 WO2021091359 A1 WO 2021091359A1
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seq
antibody
cancer
epitope
lrig
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French (fr)
Korean (ko)
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김정호
김범석
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Good T Cells Inc
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Good T Cells Inc
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Priority to US17/758,550 priority Critical patent/US20250243272A1/en
Priority to AU2020380072A priority patent/AU2020380072B2/en
Priority to CA3156600A priority patent/CA3156600A1/en
Priority to BR112022008730A priority patent/BR112022008730A2/pt
Priority to EP20885858.9A priority patent/EP4056586A4/en
Priority to JP2022525129A priority patent/JP2023500248A/ja
Priority to CN202080077856.2A priority patent/CN114929733A/zh
Publication of WO2021091359A1 publication Critical patent/WO2021091359A1/ko
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • A—HUMAN NECESSITIES
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6849—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
    • A—HUMAN NECESSITIES
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    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6854—Immunoglobulins
    • G01N33/6857—Antibody fragments
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/34—Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
    • G—PHYSICS
    • G01—MEASURING; TESTING
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    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3

Definitions

  • the present invention relates to an epitope of Lrig-1 (leucine-rich and immunoglobulin-like domains 1) protein, which is an antigen present on the surface of regulatory T cells, and an antibody or antigen-binding fragment specifically binding thereto.
  • Lrig-1 leucine-rich and immunoglobulin-like domains 1
  • the most important trait in all normal individuals is that they do not react detrimentally to the antigenic substances that make up their own, while the ability to recognize and eliminate non-self antigens. .
  • the non-response of a living body to its own antigen is called immunologic unresponsiveness or tolerance.
  • Self-tolerance occurs by removing lymphocytes that may have specific receptors for self-antigens, or by inactivating their ability to respond after contact with self-antigens.
  • an immune response to the self-antigen occurs, and a disease resulting from this is called an autoimmune disease.
  • the regulatory T cells play an important role in naturally preventing the occurrence of excessive inflammation and immune responses, but when autoimmune diseases and chronic inflammatory diseases occur, the function and number of regulatory T cells are significantly reduced. Reported. Therefore, in the case of patients with immune diseases and inflammatory diseases, it is important that regulatory T cells are produced at a normal level, which can be one of the treatments for these diseases.
  • CDR complementarity determining regions
  • An object of the present invention is to provide an epitope of Lrig-1 (leucine-rich and immunoglobulin-like domains 1) protein present on the surface of regulatory T cells (Treg cells).
  • Another object of the present invention is to provide an antibody or antigen-binding fragment capable of specifically binding to the epitope.
  • Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer comprising an antibody or antigen-binding fragment capable of specifically binding to the epitope as an active ingredient.
  • Another object of the present invention is to provide an antibody-drug conjugate (ADC) in which, for example, an anticancer agent is combined with the antibody and a drug.
  • ADC antibody-drug conjugate
  • Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer comprising an antibody-drug conjugate as an active ingredient.
  • Another object of the present invention is an antibody or antigen-binding fragment capable of specifically binding to the epitope; And it is to provide a method of preventing or treating cancer using an antibody-drug conjugate.
  • the present invention relates to an epitope of Lrig-1 (leucine-rich and immunoglobulin-like domains 1) protein or an antibody or antigen-binding fragment that specifically binds to the epitope.
  • Lrig-1 leucine-rich and immunoglobulin-like domains 1
  • the "Lrig-1 protein” is a transmembrane protein present on the surface of a regulatory T cell, and the extracellular or lumen side leucine-rich repeat (LRR) and three immune body-like domains (immunoglobulin-like domains), transmembrane sequence and cytoplasmic tail.
  • the LRIG gene family includes LRIG1, LRIG2 and LRIG3, and the amino acids constituting each family are highly conservative.
  • the LRIG1 gene is highly expressed in normal skin and can be expressed in basal and hair follicle cells to regulate the proliferation of epithelial stem cells.
  • the Lrig-1 protein may be a protein present in humans or mice, but is not limited thereto.
  • the Lrig-1 protein may be a Lrig-1 protein derived from mammals, including primates such as humans and monkeys, rodents such as mice and rats, and the like.
  • the Lrig-1 protein may be a human-derived Lrig-1 protein represented by SEQ ID NO: 1, which may be encoded by a nucleic acid sequence represented by SEQ ID NO: 2, but is not limited thereto.
  • the Lrig-1 protein may be a mouse-derived Lrig-1 protein represented by SEQ ID NO: 3, which may be encoded by a nucleic acid sequence represented by SEQ ID NO: 4, but is not limited thereto.
  • the Lrig-1 protein may be an extracellular domain of the Lrig-1 protein, but is not limited thereto.
  • the Lrig-1 extracellular domain of the present invention may be an extracellular domain of Lrig-1 protein derived from mammals, including primates such as humans and monkeys, rodents such as mice and rats, and the like.
  • the extracellular protein of the Lrig-1 protein may be an extracellular domain of the Lrig-1 protein derived from human or mouse, but is not limited thereto.
  • the extracellular domain of the Lrig-1 protein may be represented by SEQ ID NO: 5 corresponding to the 35th to 794th amino acid sequence of the human-derived Lrig-1 protein, but is not limited thereto.
  • the extracellular domain of the Lrig-1 protein may be represented by SEQ ID NO: 6 corresponding to the 35th to 794th amino acid sequence of the mouse-derived Lrig-1 protein, but is not limited thereto.
  • an epitope of the Lrig-1 protein some amino acids of the Lrig-1 protein represented by SEQ ID NOs: 1, 3, 5, and 5, for example, 2 to 50; 6 to 45; Or 10 to 44 amino acids, but is not limited thereto.
  • an amino acid sequence represented by SEQ ID NOs: 7 to 70 it provides an epitope comprising at least one polypeptide selected from the group consisting of polypeptides consisting of.
  • the epitope of the Lirg-1 protein may be an epitope including a polypeptide consisting of an amino acid sequence represented by SEQ ID NO: 71 or SEQ ID NO: 72, but is not limited thereto.
  • the epitope of the present invention may be a conformational epitope.
  • the "stereoscopic epitope" of the present invention is composed of a discontinuous amino acid sequence, unlike a one-dimensional linear epitope composed of a continuous sequence. These conformational epitopes react with the three-dimensional structure of the antibody antigen-binding site.
  • nucleic acid molecule encoding the epitope provided by the present invention is provided.
  • the nucleic acid molecule of the present invention includes all nucleic acid molecules in which the amino acid sequence of the polypeptide provided by the present invention is translated into a polynucleotide sequence as known to those skilled in the art. Therefore, various polynucleotide sequences can be prepared by ORF (open reading frame), all of which are also included in the nucleic acid molecule of the present invention.
  • ORF open reading frame
  • an expression vector into which the isolated nucleic acid molecule provided in the present invention is inserted is provided.
  • the "vector” is a nucleic acid molecule capable of transporting another nucleic acid to which a nucleic acid molecule is linked.
  • a vector which refers to circular double-stranded DNA to which additional DNA segments can be ligated.
  • a phage vector Another type of vector is a viral vector, in which additional DNA segments can be ligated to the viral genome.
  • Certain vectors are capable of autonomous replication in the host cell into which they have been introduced (eg, bacterial vectors are episomal mammalian vectors that have a bacterial origin of replication).
  • vectors e.g., non-episomal mammalian vectors
  • vectors can be incorporated into the host cell's genome as it enters the host cell, through which it replicates with the host genome.
  • some vectors are capable of directing the expression of genes to which they are linked in the operating dimension.
  • Such vectors are referred to herein as “recombinant expression vectors” or simply “expression vectors”.
  • expression vectors useful in recombinant DNA techniques often exist in the form of plasmids.
  • plasmid and vector may be used interchangeably because plasmid is the most commonly used form among vectors.
  • the expression vector in the present invention include commercially widely used pCDNA vector, F, R1, RP1, Col, pBR322, ToL, Ti vector; Cosmid; Phages such as lambda, lambdoid, M13, Mu, p1 P22, Q ⁇ , T-even, T2, T3, and T7; It may be selected from the group consisting of plant viruses, but is not limited thereto, and all expression vectors known as expression vectors to those skilled in the art can be used in the present invention, and when selecting an expression vector, it depends on the properties of the target host cell.
  • the vector When the vector is introduced into the host cell, it may be performed by calcium phosphate transfection, viral infection, DEAE-dextran control transfection, lipofectamine transfection, or electroporation, but is not limited thereto, and those skilled in the art use.
  • An introduction method suitable for an expression vector and a host cell can be selected and used.
  • the vector contains one or more selection markers, but is not limited thereto, and may be selected according to whether or not the product is produced using a vector that does not contain a selection marker.
  • the selection of the selection marker is selected by the host cell of interest, and the present invention is not limited thereto because it uses a method already known to those skilled in the art.
  • a tag sequence may be inserted and fused onto the expression vector.
  • the tags include, but are not limited to, a hexa-histidine tag, a hemagglutinin tag, a myc tag, or a flag tag, and tags that facilitate purification known to those skilled in the art can all be used in the present invention.
  • a host cell line transformed with the expression vector provided by the present invention is provided.
  • the "host cell” includes individual cells or cell cultures that may be recipients of the vector(s) for incorporation of the polypeptide insert or were recipients.
  • a host cell includes the progeny of a single host cell, which progeny may not necessarily be completely identical (morphologically or in genomic DNA complement) to the original parental cell due to natural, accidental or deliberate mutations.
  • Host cells include cells transformed in vivo with the polypeptide(s) herein.
  • the host cells may include cells of mammalian, plant, insect, fungal or cellular origin, and include bacterial cells such as E. coli, Streptomyces, and Salmonella typhimurium; Fungal cells such as yeast cells and Pichia pastoris; Insect cells such as Drozophila and Spodoptera Sf9 cells; CHO (Chinese hamster ovary cells), SP2/0 (mouse myeloma), human lymphoblastoid, COS, NSO (mouse myeloma), 293T, Bow melanoma cells, HT-1080, BHK (Baby Hamster Kidney cells), HEK (Human Embryonic Kidney cells) or PERC.6 (Human Retinal Cells) animal cells; Alternatively, it may be a plant cell, but the present invention is not limited thereto, and any cell that can be used as a host cell line known to those skilled in the art may be used.
  • bacterial cells such as E. coli,
  • the transformation method of the present invention is an arbitrary method of injecting a vector of interest into the host cell, and any known method capable of injecting the vector into the host cell may be included.
  • a method using CaCl 2 Electroporation method, microinjection method, calcium phosphate precipitation method, electroporation method, liposome-mediated transfection method, DEAE-dextran treatment method, transformation method using gene bombadment and virus, etc., but are limited thereto. It is not.
  • an antibody or antigen-binding fragment that specifically binds to the epitope of the present invention.
  • the "antibody” refers to a protein molecule that acts as a receptor for specifically recognizing an antigen, including immunoglobulin molecules that are immunologically reactive with a specific antigen.
  • the antigen may be Lrig-1 protein present on the surface of regulatory T cells. Preferably, it may specifically recognize the leucine rich region or immunoglobulin-like domain of the Lrig-1 protein, but is not limited thereto.
  • the "immunoglobulin” has a heavy chain and a light chain, and each of the heavy and light chains includes a constant region and a variable region.
  • the variable regions of the light and heavy chains include three multivariable regions and four framework regions, referred to as complementarity determining regions (hereinafter referred to as "CDR").
  • CDRs mainly play a role in binding to the epitope of the antigen.
  • the CDRs of each chain are typically referred to as CDR1, CDR2 and CDR3 sequentially starting from the N-terminus, and are also identified by the chain in which the particular CDR is located.
  • the "full-length antibody” is a structure having two full-length light chains and two full-length heavy chains, each light chain is connected by a heavy chain and a disulfide bond, IgA, IgD, IgE, IgM, and IgG.
  • the IgG is a subtype, and includes IgG1, IgG2, IgG3 and IgG4.
  • the "antigen-binding fragment” refers to a fragment having an antigen-binding function
  • examples of the antigen-binding fragment include (1) the variable region of the light chain (VL) and the variable region of the heavy chain (VH) and the light chain.
  • a proteolytic enzyme such as papain or pepsin
  • the antibody is a monoclonal antibody, a polyclonal antibody, a chimeric antibody, a humanized antibody, a human antibody, a bivalent, Bispecific molecule, minibody, domain antibody, bispecific antibody, antibody mimic, unibody, diabody, triabody, tetrabody, or It may be a fragment thereof, but is not limited thereto.
  • the "monoclonal antibody” refers to an antibody molecule having a single molecular composition obtained from substantially the same antibody population, and exhibits a single binding specificity and affinity for a specific epitope.
  • the "chimeric antibody” is an antibody obtained by recombining the variable region of a mouse antibody and the constant region of a human antibody, and has an improved immune response compared to a mouse antibody.
  • the "humanized antibody” refers to an antibody in which the protein sequence of an antibody derived from a non-human species is modified to resemble an antibody variant naturally produced in humans.
  • the humanized antibody can be prepared by recombining a mouse-derived CDR with a human antibody-derived FR to produce a humanized variable region, and then recombining it with a desired human antibody constant region to prepare a humanized antibody.
  • the "binding" or “specific binding” refers to the affinity of the antibody or antibody composition of the present application for an antigen.
  • “Specific binding” in antigenic antibody binding can be distinguished from non-specific background binding, typically when the dissociation constant (Kd) is less than 1x10 -5 M or less than 1x10 -6 M or less than 1x10 -7 M.
  • Specific binding can be detected by methods known in the art, such as ELISA, surface plasmon resonance (SPR), immunoprecipitation, coprecipitation, etc., and non-specific binding and specific binding Include appropriate controls that can be distinguished.
  • the antibody or antigen-binding fragment of the present invention may exist as a multimer, such as a dimer, a trimer, a tetramer, or a pentamer including at least a part of the antigen-binding ability of the monomer.
  • Such multimers also include homomultimers, or heteromultimers. Since the antibody multimer contains a large number of antigen-binding sites, it has excellent antigen-binding ability compared to monomers.
  • Antibody multimers are also easy to produce bifunctional, trifunctional, and tetrafunctional antibodies.
  • multifunctional refers to an antibody or antigen-binding fragment having two or more activities or functions (eg, antigen-binding ability, enzyme activity, ligand or receptor-binding ability).
  • the antibody of the present invention is Polypeptides having enzymatic activity such as luciferase, acetyltransferase, galactosidase, and the like can be combined.
  • Multifunctional antibodies also include antibodies in multivalent or bispecific, trispecific, etc. forms.
  • composition comprising an antibody or antigen-binding fragment as an active ingredient
  • the antibody according to the present invention specifically binds to an epitope containing a polypeptide represented by any one of SEQ ID NOs: 7 to 72 present on regulatory T cells to activate or maintain the function of the regulatory T cells. Let it be; Or by inhibiting, it is possible to prevent or treat cancer by regulating the activity of effector T cells.
  • the “cancer” of the present invention represents or refers to a physiological condition characterized by unregulated cell growth, typically in mammals.
  • the cancer to be prevented, improved or treated may be a solid tumor composed of a mass generated by abnormal cell growth in a solid organ, and gastric cancer, liver cancer, or Gliocytoma, ovarian cancer, colon cancer, head and neck cancer, bladder cancer, kidney cell cancer, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma, or lung cancer, but are not limited thereto.
  • the antibody or antigen-binding fragment provided by the present invention provides an antibody-drug conjugate (Antibody-Drug Conjugate, ADC) comprising a drug.
  • ADC Antibody-Drug Conjugate
  • the "antibody-drug conjugate (ADC)" refers to a form in which a drug and an antibody are chemically linked without reducing the biological activity of the antibody and the drug.
  • the antibody-drug conjugate is a form in which a drug is bound to an amino acid residue at the N-terminus of the heavy and/or light chain of an antibody, specifically, a drug at the N-terminus, an ⁇ -amine group of the heavy and/or light chain of the antibody It refers to this combined form.
  • the "drug” may mean any substance having a specific biological activity in cells, which is a concept including DNA, RNA, or peptide.
  • the drug may be in a form including a reactive group capable of crosslinking by reacting with an ⁇ -amine group, and also includes a form in which a linker including a reactive group capable of crosslinking by reacting with an ⁇ -amine group is connected.
  • an example of a reactive group capable of crosslinking by reacting with the ⁇ -amine group is not particularly limited as long as it can be crosslinked by reacting with an ⁇ -amine group at the N-terminus of the heavy or light chain of an antibody. All types that react with known amine groups are included. Examples include Isothiocyanate, Isocyanates, Acyl azide, NHS ester, Sulfonyl chloride, Aldehyde, Glyoxal, Epoxide, Oxirane, Carbonate, Aryl halide, Imidoester, Carbodiimide, Anhydride and Fluorophenyl ester), but is not limited thereto.
  • the antibody-drug conjugate is an epitope of the present invention, that is, Lrig-1 protein; Or an epitope comprising a polypeptide consisting of a partial amino acid sequence of the extracellular domain of the Lrig-1 protein; An epitope comprising a polypeptide consisting of an amino acid sequence represented by Formula 1 above; Or an antibody or antigen-binding fragment that specifically binds to an epitope comprising a polypeptide represented by the amino acid sequence of any one of SEQ ID NOs: 48 to 113, wherein the drug is used to treat diseases targeted by the Lrig-1 antibody.
  • Any drug that can be included may be included, and for example, it may be a drug capable of treating an anticancer agent capable of treating cancer, but is not limited thereto.
  • the anticancer agent may be included without limitation as long as it is a drug used for the prevention, improvement or treatment of cancer, for example, nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, nerati Nip, Lapatinib, Gefitinib, Vandetanib, Nirotinib, Semasanib, Bosutinib, Akcitinib, Cediranib, Restaurtinib, Trastuzumab, Gefitinib, Bortezomib, Sunitinib, Carbople Latin, sorafenib, bevacizumab, cisplatin, cetuximab, viscum album, asparaginase, tretinoin, hydroxycarbamide, dasatinib, estramustine, gemtuzumab ozogamycin, ibritumomab tusetan
  • composition comprising an antibody-drug conjugate (ADC) as an active ingredient
  • the antibody or antigen-binding fragment provided by the present invention provides a pharmaceutical composition for the prevention or treatment of cancer comprising an antibody-drug conjugate (Antibody-Drug Conjugate, ADC) as an active ingredient.
  • an antibody-drug conjugate Antibody-Drug Conjugate, ADC
  • the antibody or antigen-binding fragment contained as an active ingredient in the pharmaceutical composition;
  • the antibody-drug conjugate to which the drug is bound specifically binds to an epitope comprising a polypeptide represented by any one of SEQ ID NOs: 7 to 72, thereby inhibiting the function of the regulatory T cells, and inhibiting the function of the effector T cells.
  • cancer By maintaining or increasing activity, cancer can be treated very effectively.
  • the cancer may be a solid tumor consisting of a mass caused by abnormal cell growth in a solid organ, and as a specific example, gastric cancer, liver cancer, gliocytoma, ovarian cancer, depending on the site of the solid organ.
  • gastric cancer gastric cancer
  • liver cancer gliocytoma
  • ovarian cancer depending on the site of the solid organ.
  • Colon cancer, head and neck cancer, bladder cancer, kidney cell cancer, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma, or lung cancer but are not limited thereto.
  • prevention may include, without limitation, any action of blocking, suppressing or delaying the symptoms of a disease using the pharmaceutical composition of the present invention.
  • treatment may include, without limitation, any action that improves or benefits the symptoms of a disease by using the pharmaceutical composition of the present invention.
  • the pharmaceutical composition may be in the form of capsules, tablets, granules, injections, ointments, powders, or beverages, and the pharmaceutical composition may be characterized in that it is intended for humans.
  • the pharmaceutical composition is not limited thereto, but each is formulated in the form of oral dosage forms such as powders, granules, capsules, tablets, aqueous suspensions, external preparations, suppositories, and sterile injection solutions according to conventional methods.
  • the pharmaceutical composition of the present invention may contain a pharmaceutically acceptable carrier.
  • Pharmaceutically acceptable carriers can be used as binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, coloring agents, flavoring agents, etc. for oral administration, and buffers, preservatives, and painlessness in the case of injections. Agents, solubilizers, isotonic agents, stabilizers, etc.
  • the formulation of the pharmaceutical composition of the present invention can be prepared in various ways by mixing with a pharmaceutically acceptable carrier as described above.
  • a pharmaceutically acceptable carrier as described above.
  • it when administered orally, it can be prepared in the form of tablets, troches, capsules, elixir, suspension, syrup, wafers, etc., and in the case of injections, it can be prepared in unit dosage ampoules or multiple dosage forms. have.
  • it can be formulated as a solution, suspension, tablet, capsule, sustained-release preparation, and the like.
  • suitable carriers, excipients and diluents for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , Methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil.
  • fillers, anti-coagulants, lubricants, wetting agents, flavoring agents, emulsifiers, preservatives, and the like may additionally be included.
  • the route of administration of the pharmaceutical composition is not limited thereto, but oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, Includes sublingual or rectal. Oral or parenteral administration is preferred.
  • the "parenteral" includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or injection techniques.
  • the pharmaceutical composition of the present invention may also be administered in the form of suppositories for rectal administration.
  • the pharmaceutical composition of the present invention depends on several factors, including the activity of the specific compound used, age, weight, general health, sex, formulation, time of administration, route of administration, excretion rate, drug formulation, and the severity of the specific disease to be prevented or treated. It may vary in various ways, and the dosage of the pharmaceutical composition varies depending on the patient's condition, weight, degree of disease, drug form, route and duration of administration, but may be appropriately selected by those skilled in the art, and 0.0001 to 50 mg/kg per day Alternatively, it may be administered at 0.001 to 50 mg/kg. Administration may be administered once a day, or may be divided several times. The above dosage does not limit the scope of the present invention in any way.
  • the pharmaceutical composition according to the present invention may be formulated as a pill, dragee, capsule, liquid, gel, syrup, slurry, or suspension.
  • the antibody or antigen-binding fragment according to the present invention relates to a method for preventing or treating cancer comprising administering an Antibody-Drug Conjugate (ADC) to an individual.
  • ADC Antibody-Drug Conjugate
  • the “cancer” of the present invention represents or refers to a physiological condition characterized by unregulated cell growth, typically in mammals.
  • the cancer to be prevented, improved or treated may be a solid tumor composed of a mass generated by abnormal cell growth in a solid organ, and gastric cancer, liver cancer, or Gliocytoma, ovarian cancer, colon cancer, head and neck cancer, bladder cancer, kidney cell cancer, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma, or lung cancer, but are not limited thereto.
  • the antibody or antigen-binding fragment of the present invention specifically binds to an epitope comprising a polypeptide represented by any one of the amino acid sequences of SEQ ID NOs: 7 to 72, thereby inhibiting the function of the regulatory T cells, and maintaining or increasing the activity of the effector T cells. So that cancer can be treated very effectively.
  • the "individual” is an individual suspected of developing cancer, and the suspected individual means a mammal, including mice, livestock, etc., including humans who have or may develop the disease.
  • Individuals treatable with the antibody or antibody-drug conjugate of the invention are included without limitation.
  • the methods of the present invention may comprise administering an antibody or antibody-drug conjugate in a pharmaceutically effective amount.
  • the appropriate total daily use amount may be determined by the treating physician within the range of correct medical judgment, and may be administered once or in several divided doses.
  • a specific therapeutically effective amount for a specific patient is a specific composition, including the type and degree of reaction to be achieved, whether other agents are used in some cases, the patient's age, weight, general health condition, It is preferable to apply differently according to various factors including sex and diet, administration time, administration route and secretion rate of the composition, treatment period, drugs used with or concurrently with the specific composition, and similar factors well known in the medical field.
  • the method of preventing or treating cancer may be a combination therapy further comprising administering a compound or substance having therapeutic activity against one or more cancer diseases.
  • the "combination" should be understood to represent simultaneous, individual or sequential administration.
  • the interval between administration of the second component should be such that the beneficial effect of the combination is not lost.
  • the dosage of the antibody or antibody-drug conjugate may be about 0.0001 ⁇ g to 500 mg per 1 kg of the patient's body weight, but is not limited thereto.
  • the antibody or antigen-binding fragment that specifically binds to the epitope of the Lrig-1 protein according to the present invention specifically binds to the epitope of the present invention present on the regulatory T cell to inhibit the function of the regulatory T cell, and an effector By maintaining or increasing the activity of T cells, it can be used very efficiently in the prevention, improvement or treatment of cancer.
  • FIG 1 shows the structure of the Lrig-1 protein according to an embodiment of the present invention.
  • FIG. 2 shows the structure of the Lrig-1 protein according to an embodiment of the present invention.
  • FIG. 3 shows the level of expression of Lrig-1 mRNA according to an embodiment of the present invention.
  • Figure 4 shows the level of expression of Lrig-1 mRNA according to an embodiment of the present invention.
  • FIG. 5 shows the level of expression of Lrig-1 mRNA according to an embodiment of the present invention.
  • FIG. 6 shows the level of expression of Lrig-1, Lrig-2, and Lrig-3 mRNA according to an embodiment of the present invention.
  • FIG. 7 shows a result of comparing the expression levels of Lrig-1 protein in regulatory T cells and non-regulated T cells according to an embodiment of the present invention.
  • FIG 8 shows the expression of Lrig-1 protein on the surface of regulatory T cells according to an embodiment of the present invention.
  • Lrig-1 protein GTC210-01, GTC210-02, GTC210-03, GTC210-04, GTC110-01, GTC110-02, GTC110-03 and GTC110
  • -04 regulatory mechanisms of Lrig-1 protein-induced Stat3 phosphorylation in T cells were analyzed.
  • FIG. 11 shows a design diagram of a cancer treatment experiment using a monoclonal antibody specific for Lrig-1 protein in an embodiment of the present invention.
  • FIG. 12 shows the effect of cancer treatment using monoclonal antibodies (GTC110-01, GTC110-02, GTC110-03 and GTC110-04) specific to Lrig-1 protein in an embodiment of the present invention.
  • 16 and 17 show the results of covalent labeling MS using chymotrypsin in the extracellular domain of the Lirg-1 protein according to an embodiment of the present invention.
  • FIG. 22 shows the results of cross-linked MS using chymotrypsin in the extracellular domain of the Lirg-1 protein according to an embodiment of the present invention.
  • 25 shows an epitope site to which the GTC110-04 antibody specifically binds in the extracellular domain of the Lrig-1 protein according to an embodiment of the present invention.
  • the present invention relates to an epitope of Lrig-1 (leucine-rich and immunoglobulin-like domains 1) protein or an antibody or antigen-binding fragment that specifically binds to the epitope.
  • Lrig-1 leucine-rich and immunoglobulin-like domains 1
  • the extracellular domain of the Lrig-1 protein may be represented by SEQ ID NO: 5 corresponding to the 35th to 794th amino acid sequence of the human-derived Lrig-1 protein, but is not limited thereto.
  • the extracellular domain of the Lrig-1 protein may be represented by SEQ ID NO: 6 corresponding to the 35th to 794th amino acid sequence of the mouse-derived Lrig-1 protein, but is not limited thereto.
  • T cell subtypes Th0, Th1, Th2, Th17 and iTreg were prepared. Unlike the naturally isolated nTreg, iTreg refers to cells artificially inducing differentiation in a medium containing the following composition.
  • T cells The subtype of T cells is RPMI1640 (Invitrogen Gibco, Grand Island, NY) containing 10% fetal bovine serum (FBS; hyclone, logan, UT) after separating naive T cells obtained from the spleen of mice.
  • FBS fetal bovine serum
  • Table 1 Each of the components of Table 1 below was further included in the nutrient medium, and differentiation was induced into each cell through culture for 72 hours in an incubator at 37°C and 5% CO 2.
  • the three-dimensional structure of the extracellular domain of the Lrig-1 protein was predicted.
  • LRR1 to LRR15 were present in the Lrig-LRR domain (amino acid sequence 41 to 494) of the extracellular domain of the Lrig-1 protein.
  • LRR domains consisted of 23 to 27 amino acids, and 3 to 5 leucines were present.
  • three immunoglobulin-like domains were present in amino acid sequences 494 to 781 of the Lrig-1 protein among the extracellular domains of the Lrig-1 protein.
  • Lrig-1 protein can act as a specific biomarker for regulatory T cells.
  • CD4 + T cells were isolated from the spleen of mice using a magnet-activated cell sorting (MACS) through CD4 beads. Thereafter, control T (CD4 + CD25 + T) cells and non-regulated T (CD4 + CD25 - T) cells were isolated using a fluorescent active cell sorter (FACS) using a CD25 antibody.
  • FACS fluorescent active cell sorter
  • mRNA was extracted using Trizol, and for genomic RNA, gDNA was removed by a protocol provided by the manufacturer using a gDNA extraction kit (Qiagen). . The mRNA from which gDNA was removed was synthesized as cDNA through the BDsprint cDNA synthesis kit (Clonetech).
  • the real-time polymerase chain reaction was performed under the conditions of 40 cycles at 95°C for 3 minutes, 61°C for 15 seconds, and 72°C for 30 seconds according to the protocol provided by the manufacturer using SYBR Green (Molecular Probes). Primers were used, and relative gene expression levels were calculated using the ⁇ CT method, and normalized using HPRT, and the results are shown in FIGS. 3 to 6.
  • Lrig-1 is 18.1 times higher in regulatory T (CD4 + CD25 + T) cells than in non-regulated T (CD4 + CD25 -T) cells. This was about a 10-fold higher level of expression compared to Lag3 and Ikzf4, which are known regulatory T cell markers.
  • Lrig-1 was the highest among Lrig-1, Lrig-2, and Lrig-3 corresponding to the Lrig family.
  • the Lrig-1 protein according to the present invention is specifically expressed in regulatory T cells, particularly naturally occurring regulatory T cells.
  • Lrig-1 protein expressed from Lrig-1 mRNA was specifically expressed only in regulatory T cells.
  • a FOXP3-RFP-injected (Knock-in) mouse with RFP (Red fluorescence protein) bound to the FOXP3 promoter, which is a regulatory T cell-specific transcription factor a magnet-activated cell sorter (magnet-activated) from the spleen of the mouse to CD4 beads.
  • CD4 + T cells were isolated using cell sorting; MACS). Thereafter, using the RFP protein, regulated T (CD4 + RFP + T) cells and non-regulated T (CD4 + RFP - T) cells were isolated and obtained through a fluorescent active cell sorter (FACS). Each of the cells was stained with the purchased Lrig-1 antibody and the negative control isotype, and the expression level of Lrig-1 was measured with a fluorescently active cell sorter, and the results are shown in FIG. 7.
  • the Lrig-1 protein according to the present invention is specifically expressed in regulatory T cells.
  • the Lrig-1 protein In order for the Lrig-1 protein to be a target for cell therapy, it must be expressed on the surface of regulatory T cells for more effective target treatment, so it was confirmed whether the Lrig-1 protein was expressed on the surface.
  • Each of the differentiated T cell subtypes of Preparation Example 1 was stained with anti-CD4-APC and anti-Lrig-1-PE antibodies, and the surface of each cell using a fluorescence-activated cell sorter (FACS).
  • FACS fluorescence-activated cell sorter
  • the Lrig-1 protein according to the present invention is not only specifically expressed in regulatory T cell (Treg) cells, but is particularly highly expressed on the surface of regulatory T cells.
  • An antibody specific for the Lrig-1 protein according to the present invention was prepared. This antibody was not produced by specifying a specific antigenic determinant, and an antibody capable of binding to the Lrig-1 protein at any site was produced.
  • cells expressing the Lrig-1 protein were prepared. More specifically, after cutting the DNA fragment and pcDNA (hygro) corresponding to SEQ ID NO: 2 with a cleavage enzyme, incubating at 37°C for ligation, and inserting the DNA sequence of the Lrig-1 protein. The prepared pcDNA was produced. The prepared pcDNA into which SEQ ID NO: 2 was inserted was introduced into L cells through transfection, so that the Lrig-1 protein could be expressed on the surface of the L cells.
  • pcDNA hygro
  • a total of eight heavy chains and light chains were selected by selecting the sequence of the light chain and heavy chain amino acids capable of binding to Lrig-1 expressed on the cell surface in the Human scFv library.
  • the selected heavy and light chain amino acid sequences were fused with mlgG2a Fc region or human IgG1 Fc region to prepare a monoclonal antibody.
  • the sequence of the monoclonal antibody is shown in Table 3 below.
  • each of the antibodies of Preparation Examples 1 to 8 was added to L cells stably expressing Lrig-1. After binding, a secondary antibody conjugated with eFlour 670 while being able to recognize a mouse antibody was added, and the binding force of the monoclonal antibody to the Lrig-1 protein was analyzed using FACS. Is shown in FIG. 9.
  • Lrig-1 protein-specific monoclonal antibodies (GTC210-01, GTC210-02, GTC210-03 and GTC210-04) according to the present invention have the same level of phosphorylation of Stat3 as Th17 cells. It could be confirmed that the increase was made. On the other hand, Lrig-1 protein-specific monoclonal antibodies (GTC110-01, GTC110-02, GTC110-03 and GTC110-04) according to the present invention continue to maintain the phosphorylation of Stat3 at the same level as iTreg cells, and It was confirmed to reduce.
  • the Lrig-1 protein-specific monoclonal antibody according to the present invention inhibits the growth of various solid cancer cells and can effectively prevent, improve, or treat them.
  • Each of the extracellular domains (antigens) of the Lrig-1 protein was prepared.
  • DEPC Diethyl pyrocarbonate
  • Methanol was added to each of the samples after the reaction was terminated, and the supernatant was removed by centrifugation. Thereafter, a solution containing 8 M of urea and 0.1 M of NaCl was added, resuspended, and then chymotrypsin was added to allow the peptide bond to be broken.
  • mass spectrometry Q Exactive TM Plus Mass Spectrometer (Thermo SCIENTIFIC) was performed according to the conditions described in FIGS. 13 to 15, and mass spectrometry values of the measured antigen-antibody conjugate and the antigen were compared, and the results are shown in FIG. 16 Wow, it is shown in Table 5. Further, the measured mass spectrometric value was calculated as an antigen-antibody conjugate/antigen and shown in FIG. 17.
  • the polypeptides listed in the above results have the potential of an epitope, and in particular, SEQ ID NO: It can be seen that the polypeptide consisting of the amino acid sequence represented by 29 to 32 is more likely to be an epitope of the Lrig-1 protein against the GTC110-04 antibody.
  • the polypeptides listed in the above results have the potential of an epitope, and in particular, SEQ ID NO: 32 It can be seen that a polypeptide consisting of an amino acid sequence represented by any one of SEQ ID NOs: 40 to 42, 64, and 68 is more likely to be an epitope of the Lrig-1 protein against the GTC110-04 antibody.
  • the GTC110-04 antibody is the 46-87th in the extracellular domain of the Lrig-1 protein; Alternatively, it can be seen that it can specifically bind to a polypeptide consisting of amino acids 46-64 (circled portions in Fig. 25).
  • the antibody or antigen-binding fragment that specifically binds to the epitope of the Lrig-1 protein according to the present invention specifically binds to the epitope of the present invention present on the regulatory T cell to inhibit the function of the regulatory T cell, and an effector By maintaining or increasing the activity of T cells, it can be used very efficiently in the prevention, improvement or treatment of cancer.

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PCT/KR2020/015656 2019-11-08 2020-11-09 조절 t 세포 표면 항원의 에피토프 및 이에 특이적으로 결합하는 항체 Ceased WO2021091359A1 (ko)

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AU2020380072A AU2020380072B2 (en) 2019-11-08 2020-11-09 Epitope of regulatory T cell surface antigen, and antibody specifically binding thereto
CA3156600A CA3156600A1 (en) 2019-11-08 2020-11-09 Epitope of regulatory t cell surface antigen, and antibody specifically binding thereto
BR112022008730A BR112022008730A2 (pt) 2019-11-08 2020-11-09 Epítopo de antígeno de superfície de células t reguladoras e um anticorpo que se liga especificamente ao mesmo
EP20885858.9A EP4056586A4 (en) 2019-11-08 2020-11-09 REGULATORY T CELL SURFACE ANTIGEN EPITOPE, AND ANTIBODIES BINDING SPECIFICALLY THEREFOR
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023095801A1 (ja) * 2021-11-24 2023-06-01 レグセル株式会社 ヒト誘導性制御性ｔ細胞およびその作製方法

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102302393B1 (ko) * 2018-10-17 2021-09-16 주식회사 굳티셀 Lrig-1 단백질에 특이적인 결합 분자 및 이의 용도
CN117716238A (zh) * 2021-07-16 2024-03-15 古德T细胞有限公司 调节性t细胞表面抗原的表位及其特异性结合的抗体
KR20240115192A (ko) * 2023-01-17 2024-07-25 주식회사 굳티셀 조절 t 세포 표면 항원 및 이에 특이적으로 결합하는 항체
CN120882739A (zh) * 2023-03-10 2025-10-31 古德T细胞有限公司 调节性t细胞表面抗原的表位，以及特异性结合该表位的抗体

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20180022761A (ko) * 2018-02-26 2018-03-06 연세대학교 산학협력단 조절자 Ｔ 세포에 특이적으로 존재하는 새로운 표면단백질 Ｌｒｉｇ-1의 용도
KR20180116925A (ko) * 2017-04-18 2018-10-26 주식회사 굳티셀 암 또는 면역 질환의 예방 또는 치료용 약학 조성물
KR20180117067A (ko) * 2017-04-18 2018-10-26 주식회사 굳티셀 Lrig-1 단백질에 특이적인 결합 분자 및 이의 용도
KR20180116924A (ko) * 2017-04-18 2018-10-26 주식회사 굳티셀 면역 세포 표면 단백질의 항원 결정기인 폴리펩티드
KR20190124665A (ko) * 2018-04-26 2019-11-05 주식회사 굳티셀 신규한 융합 단백질 및 이를 포함하는 암의 예방 또는 치료용 약학적 조성물

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011066048A1 (en) * 2009-10-22 2011-06-03 Thomas Jefferson University Cell-based anti-cancer compositions and methods of making and using the same
KR101847523B1 (ko) * 2011-01-24 2018-05-28 연세대학교 산학협력단 조절자 Ｔ 세포에 특이적으로 존재하는 새로운 표면단백질 Ｌｒｉｇ-1의 용도
DE102016123859B3 (de) * 2016-12-08 2018-03-01 Immatics Biotechnologies Gmbh Neue T-Zellrezeptoren und deren Verwendung in Immuntherapie
EP3755350A4 (en) * 2018-02-23 2021-11-03 TrueBinding, Inc. TREATING CANCER BY BLOCKING THE INTERACTION OF VISTA AND HIS ASSOCIATION PARTNER

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20180116925A (ko) * 2017-04-18 2018-10-26 주식회사 굳티셀 암 또는 면역 질환의 예방 또는 치료용 약학 조성물
KR20180117067A (ko) * 2017-04-18 2018-10-26 주식회사 굳티셀 Lrig-1 단백질에 특이적인 결합 분자 및 이의 용도
KR20180116924A (ko) * 2017-04-18 2018-10-26 주식회사 굳티셀 면역 세포 표면 단백질의 항원 결정기인 폴리펩티드
KR20180022761A (ko) * 2018-02-26 2018-03-06 연세대학교 산학협력단 조절자 Ｔ 세포에 특이적으로 존재하는 새로운 표면단백질 Ｌｒｉｇ-1의 용도
KR20190124665A (ko) * 2018-04-26 2019-11-05 주식회사 굳티셀 신규한 융합 단백질 및 이를 포함하는 암의 예방 또는 치료용 약학적 조성물

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
R. K. GERSHONK. KONDO, IMMUNOLOGY, vol. 18, 1970, pages 723 - 37
See also references of EP4056586A4

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023095801A1 (ja) * 2021-11-24 2023-06-01 レグセル株式会社 ヒト誘導性制御性ｔ細胞およびその作製方法
JPWO2023095801A1 (https=) * 2021-11-24 2023-06-01
JP7572755B2 (ja) 2021-11-24 2024-10-24 レグセル株式会社 ヒト誘導性制御性ｔ細胞およびその作製方法

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AU2020380072A1 (en) 2022-05-19
BR112022008730A2 (pt) 2022-07-19
CN114929733A (zh) 2022-08-19
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