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WO2022139530A1 - Composition destinée à réduire, prévenir ou traiter des maladies auto-immunes comprenant des vésicules extracellulaires dérivées de lactobacillus sakei en tant que principe actif - Google Patents

Composition destinée à réduire, prévenir ou traiter des maladies auto-immunes comprenant des vésicules extracellulaires dérivées de lactobacillus sakei en tant que principe actif Download PDF

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WO2022139530A1
WO2022139530A1 PCT/KR2021/019780 KR2021019780W WO2022139530A1 WO 2022139530 A1 WO2022139530 A1 WO 2022139530A1 KR 2021019780 W KR2021019780 W KR 2021019780W WO 2022139530 A1 WO2022139530 A1 WO 2022139530A1
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autoimmune diseases
preventing
pharmaceutical composition
extracellular vesicles
lactobacillus sakei
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Korean (ko)
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진화섭
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Liscure Biosciences Co Ltd
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Liscure Biosciences Co Ltd
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Priority claimed from KR1020210186079A external-priority patent/KR20220091427A/ko
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]

Definitions

  • the present invention relates to a composition for improving, preventing or treating autoimmune diseases such as rheumatoid arthritis.
  • the immune system plays a role in protecting the body from invading antigens from the outside. Antigens include bacteria, viruses, toxins, cancer cells, blood and tissues of other people or animals. The immune system produces antibodies to destroy these harmful substances. However, when there is an autoimmune abnormality, the immune system cannot distinguish between healthy organs and harmful antigens, destroying normal tissues. A disease induced through this reaction is an autoimmune disease.
  • Rheumatoid arthritis a representative autoimmune disease, is an inflammatory autoimmune disease that appears multiple in multiple joints.
  • inflammatory cells such as macrophages, T-cells, B cells, and dendritic cells act excessively, causing chronic inflammation, causing joint and cartilage damage. characterized by causing pain.
  • the treatment of autoimmune reactions is performed for the purpose of controlling the autoimmune response and restoring the body's damaged immune function. For example, if there is a blood problem, a blood transfusion may be required, and if there is a problem with a bone, joint or muscle, exercise or other functional treatment may be required. Medications are also prescribed to modulate the immune system's response. Accordingly, immunosuppressive medicines prescribed include prednisone, a corticosteroid drug, cyclophosphamide, azathioprine, and tacrolimus, a nonsteroid drug. ) are known.
  • these therapeutic agents may cause side effects in the body and the therapeutic effect is not sufficient, so it is necessary to develop a new therapeutic agent for autoimmune diseases such as rheumatoid arthritis, which has excellent therapeutic effect while ensuring safety.
  • An object of the present invention is to promote tolDC induction from dendritic cells to directly inhibit the proliferation of CD4 T cells, and to promote the expression of immunosuppressive activity-inducing genes such as DC-SIGN and DCIR to prevent autoimmune diseases such as rheumatoid arthritis or To provide a therapeutic pharmaceutical composition.
  • Another object of the present invention is to promote tolDC induction from dendritic cells to directly inhibit the proliferation of CD4 T cells, and to promote the expression of immunosuppressive activity-inducing genes such as DC-SIGN and DCIR to prevent autoimmune diseases such as rheumatoid arthritis Or to provide a food composition that can be improved.
  • compositions for preventing or treating autoimmune diseases comprising Lactobacillus sakei-derived extracellular vesicles as an active ingredient.
  • the autoimmune disease is rheumatoid arthritis, multiple sclerosis, Sjogren syndrome, lupus, atopic dermatitis, rhinitis, alopecia areata, psoriasis, pemphigus , asthma, aphthous stomatitis, chronic thyroiditis, acquired aplastic anemia, Behcet's disease, Crohn's disease, silicon pneumonitis, streptococcal glomerulonephritis (PSGN), dermatomyosit is, polymyositis, It may be any one selected from autoimmune hemolytic anemia, autoimmune encephalomyelitis, myasthenia gravis, Grave's disease, and polyarteritis nodosa.
  • the pharmaceutical composition for preventing or treating autoimmune diseases may be for promoting induction of tolDC (Autologous tolerogenic dendritic cells) from dendritic cells.
  • tolDC Autologous tolerogenic dendritic cells
  • the pharmaceutical composition for preventing or treating autoimmune diseases may be for inhibiting proliferation of CD4 T cells.
  • the pharmaceutical composition for preventing or treating autoimmune diseases may be for promoting expression of one or more immunosuppressive activity-inducing genes selected from DC-SIGN and DCIR.
  • a food composition for preventing or improving autoimmune diseases comprising Lactobacillus sakei-derived extracellular vesicles as an active ingredient.
  • the pharmaceutical composition for preventing or treating autoimmune diseases comprising Lactobacillus sakei-derived extracellular vesicles as an active ingredient of the present invention promotes tolDC induction from dendritic cells to directly promote CD4 T cell proliferation. It is possible to prevent or treat autoimmune diseases such as rheumatoid arthritis by inhibiting and promoting the expression of immunosuppressive activity-inducing genes such as DC-SIGN and DCIR.
  • the food composition for preventing or improving autoimmune diseases comprising Lactobacillus sakei-derived extracellular vesicles as an active ingredient of the present invention promotes tolDC induction from dendritic cells and directly promotes proliferation of CD4 T cells. It is possible to prevent or improve autoimmune diseases such as rheumatoid arthritis by inhibiting and promoting the expression of immunosuppressive activity-inducing genes such as DC-SIGN and DCIR.
  • FIG. 1 is an electron micrograph of an extracellular vesicle derived from Lactobacillus sakeai isolated according to Example 1.
  • FIG. 2 is a result of analysis of tolDC induction in mature dendritic cells according to Experimental Example 1.
  • the pharmaceutical composition for preventing or treating autoimmune diseases of the present invention contains Lactobacillus sakei-derived extracellular vesicles as an active ingredient.
  • the Lactobacillus sakeai is not limited, but preferably Lactobacillus sakeai derived from kimchi, more preferably Lactobacillus sakeai deposited under the accession number KCTC13818BP.
  • Lactobacillus sakeai of the above accession number KCTC13818BP was deposited with the Center for Biological Resources on March 07, 2019.
  • the autoimmune disease is rheumatoid arthritis, multiple sclerosis, Sjogren syndrome, lupus, atopic dermatitis, rhinitis, alopecia areata, psoriasis, pemphigus, Asthma, aphthous stomatitis, chronic thyroiditis, acquired aplastic anemia, Behcet's disease, Crohn's disease, silicon pneumonitis, streptococcal glomerulonephritis (PSGN), dermatomyosit is, polymyositis, autologous It may be any one selected from autoimmune hemolytic anemia, autoimmune encephalomyelitis, myasthenia gravis, Grave's disease and polyarteritis nodosa, preferably rheumatoid arthritis ), multiple sclerosis, or Sjogren syndrome, and more preferably rheumatoid arthritis.
  • the pharmaceutical composition for preventing or treating autoimmune diseases may be for promoting induction of tolDC (Autologous tolerogenic dendritic cells) from dendritic cells.
  • tolDC Autologous tolerogenic dendritic cells
  • the pharmaceutical composition for preventing or treating autoimmune diseases may be for inhibiting proliferation of CD4 T cells.
  • CD4 T cells are helper T cells and refer to a group of cells that activate various immune cells. There are various Th1, Th2, Th1 7 CD4 T cells including Tfh2 cells in the aggregate called CD4 T cells.
  • the pharmaceutical composition for preventing or treating autoimmune diseases has at least one immunosuppressive activity selected from DC-SIGN (Dendritic Cell-Specific Intercellular Adhesion Molecule-3-Grabbing Non-integrin) and dendritic cell immunoreceptor (DCIR). It may be for promoting expression of an inducible gene.
  • DC-SIGN Dendritic Cell-Specific Intercellular Adhesion Molecule-3-Grabbing Non-integrin
  • DCIR dendritic cell immunoreceptor
  • DC-SIGN is a type of complement receptor and acts by using C1q as a ligand. It belongs to the C-type lectin receptor (CLR), which is a C-type lectin (CLEC) receptor, and is a receptor mainly expressed in macrophages and dendritic cells. .
  • CLR C-type lectin receptor
  • CLEC C-type lectin receptor
  • DC-SIGN and similar receptors are frequently found in macrophages located in lymph nodes or marginal zones of the spleen.
  • dendritic cells are responsible for antigen presentation to T cells, and in this process, CLRs on the surface of dendritic cells, including DC-SIGN, may be involved.
  • DC-SIGN can efficiently present antigens to CD4 T cells among CLRs.
  • dendritic cell immunoreceptor can inhibit the inflammatory response by interaction with NA2 (asialo-biantennary N-glycan), a glycoprotein on the surface of dendritic cells and bone cells.
  • NA2 asialo-biantennary N-glycan
  • extracellular vesicle refers to a vesicle having a lipid bilayer structure with a diameter ranging from 20 to 1,000 nm that is secreted into the extracellular environment through the fusion of polycystic bodies and plasma membranes in various cells.
  • the average diameter of the extracellular vesicles produced by the method of the present invention may be in the range of 20 to 500 nm, preferably 50 to 300 nm, more preferably 150 to 200 nm. .
  • Extracellular vesicles having a microdiameter within this range are called exosomes.
  • step a culture Lactobacillus sakei
  • Lactobacillus sakeai can be obtained from a stock solution of kimchi extract.
  • step b the Lactobacillus sakeai cultured medium is centrifuged to recover the culture supernatant.
  • the culture supernatant is mixed with a solution containing sodium chloride and an organic solvent and reacted, followed by centrifugation to obtain an extracellular vesicle pellet (step c).
  • the culture supernatant and the solution containing sodium chloride and the organic solvent may be mixed in a volume ratio of 1:0.5 to 1:1.5, preferably in a volume ratio of 1:0.8 to 1:1.2, more preferably the same amount can be mixed with
  • the organic solvent may be polyethylene glycol.
  • the solution containing sodium chloride and the organic solvent preferably has a sodium chloride concentration of 0.5 to 1 M.
  • centrifugation may be performed, and then the mixing and centrifugation may be repeated.
  • extracellular vesicle pellet is resuspended in extracellular vesicle extraction buffer to prepare a suspension, and the extracellular vesicles are isolated from the suspension (step d).
  • the term 'comprising as an active ingredient' means including an amount sufficient to achieve the efficacy or activity of the Lactobacillus sakeai-derived extracellular vesicles.
  • the Lactobacillus sakeai-derived extracellular vesicles in the composition of the present invention are, for example, 0.001 mg/kg or more, preferably 0.1 mg/kg or more, more preferably 10 mg/kg or more. or more, more preferably 100 mg/kg or more, still more preferably 250 mg/kg or more, and most preferably 1 g/kg or more.
  • the upper quantitative limit of the Lactobacillus Sakei-derived extracellular vesicles contained in the composition of the present invention can be selected and carried out by those skilled in the art by selecting within an appropriate range. have.
  • the pharmaceutical composition of the present invention may be prepared by using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant includes an excipient, a disintegrant, a sweetener, a binder, a coating agent, a swelling agent, a lubricant, and a lubricant. agent or flavoring agent, etc. may be used.
  • the pharmaceutical composition may be preferably formulated as a pharmaceutical composition by including one or more pharmaceutically acceptable carriers in addition to the active ingredients described above for administration.
  • Formulations of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops or injectables.
  • the active ingredient may be combined with an orally, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like.
  • suitable binders, lubricants, disintegrants and color-developers may also be included in the mixture.
  • Suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tracacanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like.
  • Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.
  • acceptable pharmaceutical carriers are sterile and biocompatible, and include saline, sterile water, Ringer's solution, buffered saline, albumin injection, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostats may be added as needed.
  • diluents, dispersants, surfactants, binders and lubricants may be additionally added to form an injectable formulation such as an aqueous solution, suspension, emulsion, etc., pills, capsules, granules or tablets.
  • the pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc., preferably oral administration.
  • a suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, administration mode, age, weight, pathological condition, food, administration time, route of administration, excretion rate, and response sensitivity of the patient, usually skilled A trained physician can readily determine and prescribe an effective dosage for the desired treatment or prevention.
  • the daily dose of the pharmaceutical composition of the present invention is 0.001-10 g/kg.
  • the pharmaceutical composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person of ordinary skill in the art to which the present invention pertains. Alternatively, it may be prepared by being introduced into a multi-dose container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or stabilizer.
  • the present invention provides a food composition for preventing or improving autoimmune diseases comprising Lactobacillus sakei-derived extracellular vesicles as an active ingredient.
  • the Lactobacillus sakeai is not limited, but preferably Lactobacillus sakeai derived from kimchi, more preferably Lactobacillus sakeai deposited under the accession number KCTC13818BP.
  • Lactobacillus Sakei-derived extracellular vesicles are the same as those described in the pharmaceutical composition for preventing or treating autoimmune diseases comprising the above-described Lactobacillus Sakei-derived extracellular vesicles as an active ingredient. shall refer to.
  • the food composition according to the present invention may be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods.
  • Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, sweets, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gums, ice cream, vitamin complexes, health supplements etc.
  • the food composition of the present invention may include not only the active ingredients, but also ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents.
  • examples of the above-mentioned carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose, oligosaccharides and the like; and polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol.
  • flavoring agents natural flavoring agents [taumatine, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.
  • stevia extract eg, rebaudioside A, glycyrrhizin, etc.
  • synthetic flavoring agents sacharin, aspartame, etc.
  • the present invention provides a health functional food comprising a food composition for preventing or improving autoimmune diseases comprising the Lactobacillus sakeai-derived extracellular vesicles as an active ingredient.
  • Health functional food is a food prepared by adding Lactobacillus sakei-derived extracellular vesicles to food materials such as beverages, teas, spices, gum, and confectionery, or by encapsulating, powdering, or suspension, etc. It means to bring a specific effect, but unlike general drugs, it has the advantage of not having side effects that may occur when taking the drug for a long period of time by using food as a raw material.
  • the health functional food of the present invention obtained in this way is very useful because it can be ingested on a daily basis.
  • Lactobacillus sakeai-derived extracellular vesicles added in such a health functional food cannot be uniformly defined as it varies depending on the type of health functional food being targeted, but it can be added within a range that does not impair the original taste of the food, It is usually in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight, based on the target food.
  • a health functional food in the form of pills, granules, tablets or capsules it is usually added in an amount of 0.1 to 100% by weight, preferably 0.5 to 80% by weight.
  • the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.
  • Lactobacillus sakeai-derived extracellular vesicles according to the present invention or a composition comprising the same as an active ingredient may be used as a feed additive or feed.
  • the composition When used as a feed additive, the composition may be 20 to 90% highly concentrated or prepared in powder or granular form.
  • the feed additives include organic acids such as citric acid, humic acid, adipic acid, lactic acid, and malic acid, phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polyphosphate), polyphenol, catechin, alpha-tocopherol, rosemary Extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, any one or more of natural antioxidants such as phytic acid may be further included.
  • the composition When used as a feed, the composition may be formulated in the form of a conventional feed, and may include common feed ingredients together.
  • the feed additive and feed include grains such as milled or crushed wheat, oats, barley, corn and rice; plant protein feeds, such as feeds based on rape, soybean, and sunflower; animal protein feeds such as blood meal, meat meal, bone meal and fish meal; Sugar and dairy products, for example, may further include dry ingredients made of various powdered milk and whey powder, and may further include nutritional supplements, digestion and absorption enhancers, growth promoters, and the like.
  • the feed additive may be administered alone or in combination with other feed additives in an edible carrier.
  • the feed additive can be easily administered to the animal as a top dressing, directly mixing them with animal feed, or in an oral formulation separate from the feed.
  • a pharmaceutically acceptable edible carrier as well known in the art, and may be prepared into an immediate release or sustained release formulation.
  • Such edible carriers may be solid or liquid, for example cornstarch, lactose, sucrose, soybean flakes, peanut oil, olive oil, sesame oil and propylene glycol.
  • the feed additive may be a tablet, a capsule, a powder, a troche or a sugar-containing tablet, or a top dressing in a microdispersed form.
  • the feed additive may be in the form of a gelatin soft capsule, or a syrup, suspension, emulsion, or solution.
  • the feed additives and feed may contain auxiliary agents, for example, preservatives, stabilizers, wetting or emulsifying agents, solution accelerators, and the like.
  • the feed additive may be used by adding to animal feed by immersion, spraying, or mixing.
  • the feed or feed additive of the present invention can be applied to a number of animal diets including mammals, poultry and fish.
  • the mammal it can be used for pigs, cattle, sheep, goats, laboratory rodents, and laboratory rodents, as well as pets (eg, dogs, cats), etc., as the poultry, chickens, turkeys, ducks, geese, pheasants, and quail, and the like, and may be used as the fish such as trout, but is not limited thereto.
  • the present invention provides the use of the Lactobacillus sakeai-derived extracellular vesicles for the manufacture of a medicament or food for preventing, treating or improving autoimmune diseases.
  • the Lactobacillus Sakeai-derived extracellular vesicles can be used for the treatment or improvement of autoimmune diseases.
  • the present invention provides a method for improving, preventing or treating an autoimmune disease comprising administering to a mammal an effective amount of an extracellular vesicle derived from Lactobacillus sakeai.
  • mammal refers to a mammal that is the subject of treatment, observation or experimentation, preferably a human.
  • the term "effective amount” refers to the amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human as conceived by a researcher, veterinarian, physician or other clinician, which is the disease in question. or an amount that induces alleviation of the symptoms of the disorder.
  • the effective amount and frequency of administration for the active ingredient of the present invention may vary depending on the desired effect.
  • the optimal dosage to be administered can be easily determined by those skilled in the art, and the type of disease, the severity of the disease, the content of active ingredients and other components contained in the composition, the type of formulation, and the age, weight, and general health of the patient It can be adjusted according to various factors including state, sex and diet, administration time, administration route and secretion rate of the composition, treatment period, and concurrently used drugs.
  • the extract for adults, it is preferable to administer the extract at a dose of 0.001 g/kg to 10 g/kg when administered once to several times a day.
  • the composition comprising Lactobacillus sakeai-derived extracellular vesicles as an active ingredient is administered via oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, or intradermal routes. It can be administered in a conventional manner.
  • Example 1 Lactobacillus sakeai-derived extracellular vesicle (P12E) isolation
  • the microorganism was named Lactobacillus sakei WIKIM0109 (Lactobacillus sakei WIKIM0109) and deposited at the Center for Biological Resources (Accession No. KCTC13818BP).
  • Lactobacillus Sakeai accesion No. KCTC13818BP
  • 0.1% of Lactobacillus Sakeai was inoculated into 30 ml of MRS broth and cultured at 30° C. for 18 hours. After incubation, the culture supernatant was recovered by first centrifugation at 3500 rpm, 4°C for 10 minutes, followed by a second centrifugation at 10,000 x g, 4°C for 20 minutes, and finally filtered through a 0.22 ⁇ m filter to remove cells. was recovered. Then, the culture supernatant and 1 M NaCl solution dissolved in 16% PEG 6000 were mixed in equal amounts and reacted at 4 ° C. for 15 hours, followed by centrifugation at 10,000 x g, 4 ° C.
  • a medium in which Lactobacillus sakeai was cultured in (1) of Example 1 was prepared.
  • Example 1 the Lactobacillus sakeai of (1) was prepared without separating the extracellular vesicles.
  • Monocytes from bone marrow cells of the femur and tibia of mice were centrifuged according to the Ficoll-Paque® density gradient centrifugation method.
  • the isolated monocytes were prepared in a 6-well plate at a concentration of 2 x 10 6 /well, and GM-CSF (Granulocyte-macrophage colony-stimulating factor) was added to a medium (RPMI) containing Fetal Bovine Serum (FBS). 20 ng/ml and 10 ng/ml of IL-4 (interleukin 4) were co-treated. After that, it was cultured for 6 days, and when 3 days had elapsed, it was replaced with a fresh medium and cytokines to obtain differentiated immature dendritic cells.
  • GM-CSF Granulocyte-macrophage colony-stimulating factor
  • the positive control LPS Lipopolysaccharide
  • the Lactobacillus sakeai of Comparative Example 2 and the Lactobacillus sakeai derived extracellular vesicles (EV) of Example 1 were inoculated with MOI (multiplicity of infection) 1 as a secondary stimulus, and co-cultured for 24 hours.
  • MOI multiplicity of infection
  • PD-L1 Programmed Death-Ligand 1
  • LPS Lactobacillus sakeai-derived extracellular vesicle-treated group
  • Dendritic cells have immune activation signal and immune suppression signal, and each signal is transmitted by various factors for homeostasis of immune response.
  • Toll-like receptor 4 TLR4
  • DC-SIGN dendritic cell-specific intercellular adhesion molecule (DC-SIGN) -3-grabbing non-integrin) and dendritic cell immunoreceptor (DCIR).
  • DC-SIGN is a C-type lectin receptor, and many ligands exist, and when co-stimulated with a toll-like receptor (TLR), it can suppress the immune response.
  • Primer information and qPCR conditions are as follows.
  • DC-SIGN expression was strongly inhibited in the mature dendritic cell treatment group (LPS group), and the Lactobacillus sakeai culture medium treatment group of Comparative Example 1 (LPS+P12 media) and Lactobacillus of Comparative Example 2
  • LPS+P12 the Lactobacillus sakeai culture medium treatment group of Comparative Example 1
  • LPS+P12E the Lactobacillus sakeai-derived extracellular vesicle-treated group
  • DC-IR expression was also strongly inhibited in the mature dendritic cell treatment group (LPS group), and the Lactobacillus sakeai culture medium treatment group (LPS+P12 media) of Comparative Example 1 and Lactobacillus sake of Comparative Example 2
  • DC-SIGN expression was slightly increased in the eye treatment group (LPS+P12), and DC-IR expression was significantly increased in the Lactobacillus sakeai-derived extracellular vesicle treatment group (LPS+P12E) of Example 1.
  • CD4 T cells Increased division of CD4 T cells is known to be involved in various inflammatory responses through activation of T cells.
  • dendritic cells cultured for 24 hours with Lactobacillus sakeai of Comparative Example 2 and dendritic cells cultured for 24 hours with the Sakeai-derived extracellular vesicles of Example 1 were prepared, and CD4 T cells were harvested from mice. It was isolated and stained with carboxyfluorescein succinimidyl ester (CFSE) for cell labeling. Thereafter, tolDC-induced cultured dendritic cells: CFSE-stained CD4 T cells were co-cultured at a ratio of 1:1 for 3 days, and the co-cultured cells were harvested after 3 days to measure the degree of decrease in CFSE.
  • the results of the CD4 T cell activity inhibitory ability analysis according to this are shown in FIG. 5 .
  • the LPS-treated dendritic cells and the co-cultured T cells showed a proliferation rate of about 90%, whereas the Lactobacillus sakeai-derived extracellular vesicles of Example 1 and dendritic cells cultured for 24 hours.
  • the proliferation of CD4 T cells was significantly reduced compared to the LPS-treated group, showing a proliferation rate of about 60%, and it was confirmed that it was suppressed to a level close to that of the dendritic cell untreated group (Non-DCs). there was.
  • the immunosuppressive ability of the Lactobacillus sakeai-derived extracellular ER was realized by directly inhibiting the activity of CD4 T cells by tolDC induced by the Lactobacillus sakeai-derived extracellular ER.
  • the Lactobacillus sakeai-derived extracellular vesicles of the present invention can promote tolDC induction of dendritic cells, and tolDC can improve rheumatoid arthritis by inhibiting the activity of CD4 T cells.
  • the above ingredients are mixed and filled in an airtight bag to prepare a powder.
  • tablets are prepared by tableting according to a conventional manufacturing method of tablets.
  • the above ingredients are mixed and filled in a gelatin capsule to prepare a capsule.
  • the content of the above components per 1 ampoule is prepared.
  • each component is added to purified water to dissolve, an appropriate amount of lemon flavor is added, the above components are mixed, purified water is added, the whole is adjusted to 100 g by adding purified water, and then filled in a brown bottle to sterilize to prepare a liquid.
  • composition ratio of the vitamin and mineral mixture is relatively suitable for granules in a preferred embodiment, but the mixing ratio may be arbitrarily modified. It can be prepared and used in the preparation of a health functional food composition according to a conventional method.
  • the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, then refrigerated. It is used to prepare the functional beverage composition of the present invention.
  • composition ratio is prepared by mixing ingredients suitable for relatively favorite beverages in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and national preferences such as demand class, demanding country, and use.
  • the pharmaceutical composition for preventing or treating autoimmune diseases comprising Lactobacillus sakei-derived extracellular vesicles as an active ingredient of the present invention promotes tolDC induction from dendritic cells to directly promote CD4 T cell proliferation. It is possible to prevent or treat autoimmune diseases such as rheumatoid arthritis by inhibiting and promoting the expression of immunosuppressive activity-inducing genes such as DC-SIGN and DCIR.
  • the food composition for preventing or improving autoimmune diseases comprising Lactobacillus sakei-derived extracellular vesicles as an active ingredient of the present invention promotes tolDC induction from dendritic cells and directly promotes proliferation of CD4 T cells. It is possible to prevent or improve autoimmune diseases such as rheumatoid arthritis by inhibiting and promoting the expression of immunosuppressive activity-inducing genes such as DC-SIGN and DCIR.

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  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

La présente invention concerne une composition pour la prévention, le traitement ou la réduction de maladies auto-immunes, contenant des vésicules extracellulaires dérivées de Lactobacillus sakei en tant que principe actif. En conséquence, la composition de la présente invention inhibe directement la prolifération des lymphocytes T CD4 en favorisant l'induction de tolDC à partir de cellules dendritiques, et favorise l'expression de gènes induisant une activité immunosuppressive tels que le DC-SIGN et le DCIR, et peut ainsi prévenir, traiter ou soulager des maladies auto-immunes telles que la polyarthrite rhumatoïde.
PCT/KR2021/019780 2020-12-23 2021-12-23 Composition destinée à réduire, prévenir ou traiter des maladies auto-immunes comprenant des vésicules extracellulaires dérivées de lactobacillus sakei en tant que principe actif Ceased WO2022139530A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR10-2020-0181541 2020-12-23
KR20200181541 2020-12-23
KR10-2021-0186079 2021-12-23
KR1020210186079A KR20220091427A (ko) 2020-12-23 2021-12-23 락토바실러스 사케아이 유래 세포밖 소포체를 유효성분으로 포함하는 자가면역질환의 개선, 예방 또는 치료용 조성물

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WO2022139530A1 true WO2022139530A1 (fr) 2022-06-30

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160060652A1 (en) * 2013-03-13 2016-03-03 Lapian's Corp Microvesicle and method for producing the same
KR20200053531A (ko) * 2017-09-08 2020-05-18 에벨로 바이오사이언시즈, 인크. 박테리아 세포외 소포
KR20200060637A (ko) * 2018-11-22 2020-06-01 (주)프로스테믹스 엑소좀 및 이의 다양한 용도
KR102123505B1 (ko) * 2019-03-26 2020-06-17 한국식품연구원 관절염 저감화 효능을 갖는 김치 유산균 락토바실러스 사케아이 wikim0109
WO2020214083A1 (fr) * 2019-04-17 2020-10-22 Biogaia Ab Microvésicules thérapeutiques de bactéries probiotiques

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160060652A1 (en) * 2013-03-13 2016-03-03 Lapian's Corp Microvesicle and method for producing the same
KR20200053531A (ko) * 2017-09-08 2020-05-18 에벨로 바이오사이언시즈, 인크. 박테리아 세포외 소포
KR20200060637A (ko) * 2018-11-22 2020-06-01 (주)프로스테믹스 엑소좀 및 이의 다양한 용도
KR102123505B1 (ko) * 2019-03-26 2020-06-17 한국식품연구원 관절염 저감화 효능을 갖는 김치 유산균 락토바실러스 사케아이 wikim0109
WO2020214083A1 (fr) * 2019-04-17 2020-10-22 Biogaia Ab Microvésicules thérapeutiques de bactéries probiotiques

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