WO2020130467A2 - Composition pharmaceutique pour la prévention ou le traitement d'une maladie médiée par les lymphocytes t de mémoire cd8+ - Google Patents

Composition pharmaceutique pour la prévention ou le traitement d'une maladie médiée par les lymphocytes t de mémoire cd8+ Download PDF

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Publication number
WO2020130467A2
WO2020130467A2 PCT/KR2019/017416 KR2019017416W WO2020130467A2 WO 2020130467 A2 WO2020130467 A2 WO 2020130467A2 KR 2019017416 W KR2019017416 W KR 2019017416W WO 2020130467 A2 WO2020130467 A2 WO 2020130467A2
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WIPO (PCT)
Prior art keywords
cell
cells
memory
treatment
disease
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PCT/KR2019/017416
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English (en)
Korean (ko)
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WO2020130467A3 (fr
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신의철
한지원
김종훈
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한국과학기술원
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Publication of WO2020130467A3 publication Critical patent/WO2020130467A3/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/13—Nucleic acids or derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/13—Amines
    • A61K31/145—Amines having sulfur, e.g. thiurams (>N—C(S)—S—C(S)—N< and >N—C(S)—S—S—C(S)—N<), Sulfinylamines (—N=SO), Sulfonylamines (—N=SO2)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • 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
    • 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
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • G01N33/5047—Cells of the immune system
    • G01N33/505—Cells of the immune system involving T-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
    • 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
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/136—Screening for pharmacological compounds
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/70—Mechanisms involved in disease identification
    • G01N2800/7042—Aging, e.g. cellular aging

Definitions

  • the present invention relates to a pharmaceutical composition for the prevention or treatment of CD8 + memory T cell mediated diseases.
  • the HIF-2 ⁇ protein of the present invention is a protein encoded by the EPAS1 gene, and has a transcription factor activity in response to a decrease in oxygen available in a hypoxic state.
  • the HIF-2 ⁇ protein may have an amino acid sequence represented by SEQ ID NO: 1, and the EPAS1 gene may be composed of a nucleotide sequence represented by SEQ ID NO: 2, but is not limited thereto.
  • the inhibitor of the present invention may include all that can inhibit the expression or activity of HIF-2 ⁇ protein, for example, at least one selected from the group consisting of oligonucleotides, antibodies, antigen-binding fragments, aptamers and chemicals It may be, but is not limited thereto.
  • the antibody of the present invention is a protein molecule containing an immunoglobulin molecule that is immunologically reactive with a specific antigen, and refers to a protein molecule that acts as an antigen receptor that specifically recognizes and reacts when a specific antigen invades the body. do.
  • the antibody of the present invention also includes special antibodies such as humanized antibodies and human antibodies, and antibodies known in the art may be included in addition to novel antibodies.
  • the antibody includes functional fragments of the antibody molecule, as well as a complete form with the full length of two heavy and two light chains, as long as it has the properties of binding specifically recognizing the HIF-2 ⁇ protein.
  • treatment of the present invention may include, without limitation, any action that improves or ameliorates symptoms caused by CD8 + memory T cell mediated disease using the pharmaceutical composition of the present invention.
  • the pharmaceutical composition of the present invention may be characterized in that it is in the form of capsules, tablets, granules, injections, ointments, powders or beverages, and the pharmaceutical composition may be characterized for humans.
  • the food composition of the present invention can exhibit the effect of preventing or improving CD8 + memory T cell mediated diseases by including the inhibitor of expression or activity of HIF-2 ⁇ protein described in the pharmaceutical composition as an active ingredient. Therefore, content related to HIF-2 ⁇ , an inhibitor of expression or activity, CD8+ memory T cell mediated disease and prevention is omitted to avoid excessive complexity of the specification according to the repeated description.
  • Food composition comprising the inhibitor of the present invention as an active ingredient may be prepared in the form of various foods, for example, beverages, gum, tea, vitamin complexes, powders, granules, tablets, capsules, confectionery, rice cakes, bread, etc. have.
  • the above components of the present invention can be used independently or in combination.
  • the ratio of the additives does not correspond to the essential elements of the present invention, but may be selected from 0.1 to about 50 parts by weight per 100 parts by weight of the food composition of the present invention, but is not limited thereto.
  • the biological sample of the present invention refers to any substance, biological body fluid, tissue or cell obtained from or derived from an individual, for example, whole blood, leukocytes, peripheral blood mononuclear cells ( peripheral blood mononuclear cells, blood, including leukocyte soft coat, plasma and serum, sputum, tears, mucus, nasal washes, Nasal aspirate, breath, urine, semen, saliva, peritoneal washings, pelvic fluids, cystic fluid , Meningeal fluid, amniotic fluid, glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, nipple aspirate, bronchial aspirate ( bronchial aspirate, synovial fluid, joint aspirate, organ secretions, cells, cell extracts or cerebrospinal fluid It does not work.
  • peripheral blood mononuclear cells peripheral blood mononuclear cells, blood, including leukocyte soft coat, plasma and serum, sputum, tears, mucus, nasal washes, Nasal
  • the step of determining the level of the present HIF-2 ⁇ protein of the present invention is a Western blot, ELISA (enzyme linked immunosorbent assay), radioimmunoassay using an agent measuring the protein level containing the antibody specific for the protein (radioimmunoassay, RIA), radioimmunodiffusion, ouchterlony immunodiffusion, rocket immunoelectrophoresis, immunoprecipitation assay, complement fixation assay, flow cytometry (FACS) and a protein chip (protein chip), and the like, but is not limited thereto.
  • the antibody of the present invention can be easily produced by a method known to those of ordinary skill in the art with reference to SEQ ID NO: 1, which is the amino acid sequence of HIF-2 ⁇ .
  • the candidate substance when the expression level of the HIF-2 ⁇ protein or the gene encoding it measured after treatment of the candidate substance is reduced compared to before the candidate substance treatment, the candidate substance is judged as a therapeutic agent for CD8 + T cell mediated disease It further comprises the steps.
  • composition for measuring aging of the present invention includes an agent that measures the level of HIF-2 ⁇ protein or a gene encoding it.
  • the contents related to the CD69 + CD103 - CD8 + memory T cells, HIF-2 ⁇ , etc. are omitted to avoid excessive complexity of the specification according to the repeated description.
  • the CD69 + CD103 - - - On the basis of being a CD8 + memory T cell populations (Subpopulation) resident tissue, the CD69 + CD103 - - The compositions according to the present invention CD8 + memory T cells from the CD69 + CD103 existing between the CD8 + memory only T cell-specific By inhibiting the expression level or activity of HIF-2 ⁇ present at a high level, it is possible to induce the apoptosis of CD69 + CD103 - CD8 + memory T cells, or to effectively suppress the function of the cells. Furthermore, through this effect, it can be very usefully applied to the prevention, improvement or treatment of CD8 + memory T cell mediated diseases.
  • FIGS. 7 to 10 show the results of confirming the types of genes specifically expressed in CD69 + CD103 - CD8 + memory T cells according to an embodiment of the present invention.
  • mononuclear cell suspensions were prepared from the tissues using a tumor dissociation kit (Miltenyi Biotec, Germany) and GentleMACS TM dissociator.
  • HLA Human leukocyte antigen
  • T which is an indicator related to terminal differentiation of T cells in the CD103 - cell subset It was confirmed that the frequency of the presence of EMRA cells, CD57 + cells and T-bet low Eomes hi cells was higher.
  • the instrument for sequencing was HiSeq2500 (Illumina, USA), and the sequencing conditions were such that RAW reads were set to conditions aligned to the reference genome (UCSC.hg38) through the HISAT2 package, and the reads sorted using the StringTie package. A copy assembly was generated, and the expression profile of the gene could be presented as a fragment per kilobase of transcript per million mapping leads.
  • CD69 + CD8 + memory T cells sorted from LSMCs of chronic HBV infected patients maintained activity and confirmed cell death
  • CD69 + CD8 + memory T cells in the CD103 LSMCs obtained from a chronic HBV infected-cell subset and CD103 + cells was confirmed TCR (T cell receptor) in a reactive subset.
  • TCR T cell receptor
  • 1 ⁇ g/mL HBVcore overlapping peptide was treated with RPMI culture containing LSMCs and 10% fetal calf serum, and cultured for 1 hour, Brefeldin A (BD Biosciences), monensin (BD Biosciences) and The CD107a antibody (PE- or FITC-conjugated) was further added, followed by further incubation for 5 hours. Then, after washing the cells in which the culture was completed, the antibody to which the fluorescence for a specific surface marker was conjugated was added and cultured.

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Abstract

La présente invention concerne une composition pour la prévention, le soulagement, ou le traitement d'une maladie médiée par les lymphocytes T de mémoire CD8+. Sur la base du fait que les lymphocytes T de mémoire CD69+ CD103- CD8+ parmi les lymphocytes T de mémoire CD8 + résidant dans le foie forment une sous-population résidente dans les tissus, la composition selon la présente invention inhibe le niveau d'expression ou l'activité de HIF-2α, qui est spécifiquement présent à un niveau d'expression élevé uniquement dans les lymphocytes T de mémoire CD69+ CD103- CD8+, ce qui a pour effet d'induire la mort des lymphocytes T de mémoire CD69+ CD103- CD8+ ou de supprimer efficacement leur fonction. En outre, avec de tels effets, la composition peut être appliquée à la prévention, au soulagement ou au traitement d'une maladie médiée par les lymphocytes T de mémoireCD8+.
PCT/KR2019/017416 2018-12-17 2019-12-10 Composition pharmaceutique pour la prévention ou le traitement d'une maladie médiée par les lymphocytes t de mémoire cd8+ WO2020130467A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020180163063A KR102225039B1 (ko) 2018-12-17 2018-12-17 Cd8+ 메모리 t 세포 매개성 질환의 예방 또는 치료용 약학 조성물
KR10-2018-0163063 2018-12-17

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WO2020130467A2 true WO2020130467A2 (fr) 2020-06-25
WO2020130467A3 WO2020130467A3 (fr) 2020-08-06

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Publication number Priority date Publication date Assignee Title
WO2023121197A1 (fr) * 2021-12-24 2023-06-29 주식회사 리버스매직 Composition et kit d'évaluation du vieillissement immunitaire

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20180049995A1 (en) 2015-03-11 2018-02-22 Peloton Therapeutics, Inc. Hif-2-alpha inhibitor polymorphs
WO2018160772A1 (fr) * 2017-02-28 2018-09-07 The United State Of America, As Represented By The Secretary, Department Of Health & Human Services Procédé de traitement de l'obésité, de la résistance à l'insuline, d'une stéatose hépatique non alcoolique comprenant une stéatohépatite non alcoolique
CN108969515A (zh) * 2018-08-01 2018-12-11 大连天星生物技术有限责任公司 HIF-2α抑制剂用于制备防治急性高原反应药物的用途
JP7440509B2 (ja) 2018-11-09 2024-02-28 ベス イスラエル デアコネス メディカル センター Ｃｄｃｐ１－標的化療法

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KR20200074545A (ko) 2020-06-25
KR102225039B1 (ko) 2021-03-10

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