KR20220154512A - A COMPOSITION FOR ACTIVATING TGF-β RECEPTOR OF DENDRITIC CELL AND A PREVENTING OR TREATING METHOD OF AUTOIMMUNE DISEASE USING THE SAME - Google Patents

Abstract

A noble synthesized compound represented by chemical formula 1 according to one aspect of the present invention can activate a TGF-β receptor of a dendritic cell to increase the immune tolerance of the dendritic cell. Furthermore, the dendritic cell with increased immune tolerance can exhibit an effect of preventing and/or treating an autoimmune disease through an immunosuppressive effect in vivo. In the chemical formula 1, each of R_1 to R_3 is independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, and substituted or unsubstituted alkynyl and n represents the number of OR_3, wherein n is an integer selected from 1 to 5 and if n is 2 or more, 2 or more pieces of OR_3 may be the same or different from each other.

Description

Composition for activating TGF-β receptors in dendritic cells and method for preventing or treating autoimmune diseases using the same

It relates to a composition for activating TGF-β receptors in dendritic cells and a method for preventing or treating autoimmune diseases using the same.

Transforming growth factor-β (TGF-β) is a representative immunosuppressive cytokine, which plays a role in maintaining immune homeostasis and is involved in recovery of inflammatory responses and protection from autoimmune diseases.

Autoimmune disease is a state in which the body's immune balance is broken, and the main cause is an excessive immune response of Th1 and Th17 cells, which are subtypes of helper T cells. This is improved by restoring immune homeostasis as cells that function in immune tolerance, such as regulatory T cells, increase.

Dendritic cells are antigen-presenting cells that are activated by delivering antigen information to T cells, and function in immune activation or immune tolerance depending on their maturity. Depending on environmental signals present around dendritic cells, maturation levels are differentiated differently, and in particular, the immunotolerant characteristics of dendritic cells are increased by TGF-β signals. In immune-tolerant dendritic cells, the expression of cell surface molecules and the secretion of inflammatory cytokines are reduced, thereby reducing the differentiation of Th1 and Th17 cells, while promoting the differentiation and proliferation of regulatory T cells.

Therefore, a new synthetic TGF-β agonist (hereinafter referred to as T74) capable of stimulating TGF-β signaling is prepared, treated with dendritic cells to produce immune-tolerant dendritic cells, and developed as a cell therapy agent for autoimmune diseases.

Korean Patent Publication No. 10-2021-0009197

One aspect is to provide a composition for activating a TGF-β receptor comprising a compound of formula 1 or a pharmaceutically acceptable salt thereof:

[Formula 1]

Wherein R ₁ to R ₃ are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, and substituted or unsubstituted alkynyl,

wherein n represents the number of OR ₃ , n is an integer selected from 1 to 5;

When n is 2 or more, OR ₃ of 2 or more is the same as or different from each other.

Another aspect is to provide a composition for activating a TGF-β receptor comprising a compound of Formula 2 or a pharmaceutically acceptable salt thereof:

[Formula 2]

Wherein R ₁ to R ₃ and R ₃ ′ are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, and substituted or unsubstituted alkynyl.

Another aspect is to provide a composition for activating a TGF-β receptor comprising a compound of Formula 3 or a pharmaceutically acceptable salt thereof:

[Formula 3]

Wherein R ₁ to R ₃ and R ₃ ′ are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, and substituted or unsubstituted alkynyl.

Another aspect is to provide a composition for activating a TGF-β receptor comprising a compound of Formula 4 or a pharmaceutically acceptable salt thereof:

[Formula 4]

.

.

Another aspect includes culturing immature dendritic cells and the composition for activating the TGF-β receptor together; and

It is to provide a method for producing dendritic cells with enhanced immunotolerance comprising culturing the cultured immature dendritic cells together with an antigen or a fragment thereof.

Another aspect is to provide dendritic cells with enhanced immune tolerance prepared according to the above production method.

Another aspect is to provide a pharmaceutical composition for preventing or treating autoimmune diseases containing dendritic cells having enhanced immune tolerance.

Another aspect is to provide a method for preventing or treating an autoimmune disease comprising administering the dendritic cells with enhanced immune tolerance to a subject in need thereof.

Another aspect is a health functional food for preventing or improving autoimmune diseases containing dendritic cells with enhanced immune tolerance,

The pharmaceutically acceptable salt is to provide a food-acceptable salt, a health functional food.

The compounds described herein may contain one or more asymmetric centers and thus may exist in various isomeric forms, eg enantiomers and/or diastereomers. For example, the compounds described herein may be in the form of individual enantiomers, diastereomers, or geometric isomers, or may be in the form of mixtures of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. Isomers can be isolated from mixtures by methods known to those skilled in the art including chiral high pressure liquid chromatography (HPLC) and formation and crystallization of chiral salts; Alternatively, the desired isomer may be prepared by asymmetric synthesis. The present invention further includes the compounds described herein as individual isomers, substantially free of other isomers, and, alternatively, as mixtures of various isomers. Furthermore, the present invention is not intended to be limited in any way by the exemplary list of substituents set forth herein.

One aspect provides a composition for activating a TGF-β receptor comprising a compound of Formula 1 or a pharmaceutically acceptable salt thereof:

[Formula 1]

Wherein R ₁ to R ₃ are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, and substituted or unsubstituted alkynyl,

wherein n represents the number of OR ₃ , n is an integer selected from 1 to 5;

When n is 2 or more, OR ₃ of 2 or more is the same as or different from each other.

The term "substituted" group has a substituent at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituents at each position are the same or different from each other. The term "substituted" is intended to include substitution with all permissible substituents of an organic compound, wherein any substituent described herein results in the formation of a stable compound. The present invention contemplates any and all combinations of these to arrive at a stable compound.

The term “alkyl” refers to a radical of a straight-chain or branched saturated hydrocarbon group having 1 to 5 carbon atoms (“C _1-5 alkyl”). In some embodiments, an alkyl group has 1 to 4 carbon atoms (“C _1-4 alkyl”). In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C _1-3 alkyl”). In some embodiments, an alkyl group has 1 to 2 carbon atoms (“C _1-2 alkyl”). In some embodiments, an alkyl group has 1 carbon atom (“C ₁ alkyl”). In some embodiments, an alkyl group has 1 to 5 carbon atoms (“C _1-5 alkyl”). Examples of C _1-5 alkyl are methyl (C ₁ ), ethyl (C ₂ ), propyl (C ₃ ) (eg n-propyl, isopropyl), butyl (C ₄ ) (eg n- butyl, tert-butyl, sec-butyl, isobutyl) and pentyl (C ₅ ) (eg n-pentyl, 3-pentanyl, amyl, neopentyl, 3-methyl-2-butanyl, tert-amyl) includes Unless otherwise specified, each instance of an alkyl group is independently unsubstituted (“unsubstituted alkyl”) or substituted with one or more substituents (eg, halogen such as F) (“substituted alkyl”).

The term “alkenyl” refers to a radical of a straight-chain or branched unsaturated hydrocarbon group having 2 to 5 carbon atoms (“C _2-5 alkenyl”). In some embodiments, an alkenyl group has 2 to 4 carbon atoms (“C _2-4 alkenyl”). In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C _2-3 alkenyl”). In some embodiments, an alkenyl group has 2 carbon atoms (“C ₂ alkenyl”). In some embodiments, an alkenyl group has 2 to 5 carbon atoms (“C _2-5 alkenyl”). Examples of C _2-5 alkenyl include ethenyl (C ₂ ), propenyl (C ₃ ) (eg 1-propenyl), butenyl (C ₄ ) (eg 1-butenyl, 2 -butenyl, 2-methyl-1-propenyl, etc.) and pentenyl (C ₅ ) (eg, 1-pentenyl, 2-pentenyl, 2-methyl-1-butenyl, 2-methyl-2 -butenyl, etc.). Unless otherwise specified, each instance of an alkenyl group is independently unsubstituted (“unsubstituted alkenyl”) or substituted with one or more substituents (eg, halogen such as F) (“substituted alkenyl”). ").

The term “alkynyl” refers to a radical of a straight-chain or branched unsaturated hydrocarbon group having 2 to 5 carbon atoms (“C _2-5 alkynyl”). In some embodiments, an alkynyl group has 2 to 4 carbon atoms (“C _2-4 alkynyl”). In some embodiments, an alkynyl group has 2 to 3 carbon atoms (“C _2-3 alkynyl”). In some embodiments, an alkynyl group has 2 carbon atoms (“C ₂ alkynyl”). In some embodiments, an alkynyl group has 2 to 5 carbon atoms (“C _2-5 alkynyl”). Examples of C _2-5 alkynyl include ethynyl (C ₂ ), propynyl (C ₃ ) (eg, 1-propynyl, 2-methyl-1-ethynyl, etc.), butynyl (C ₄ ) ( eg, 1-butynyl, 2-butynyl, etc.) and pentynyl (C ₅ ) (eg, 1-pentynyl, 2-pentynyl, 3-methyl-1-pentynyl, etc.). . Unless otherwise specified, each instance of an alkynyl group is independently unsubstituted (“unsubstituted alkynyl”) or substituted with one or more substituents (eg, halogen such as F) (“substituted alkynyl”). ").

The compound of Formula 1 or a pharmaceutically acceptable salt thereof may be derived from nature or synthesized using a known organic synthesis method.

In addition, the compound of Formula 1 or a pharmaceutically acceptable salt thereof may be a non-protein compound, a peptide, a plant-derived tissue or cell extract, a product obtained by culturing a microorganism (eg bacteria or fungi, and particularly yeast). have.

The term "salt" refers to a salt prepared using a specific compound according to one aspect and a relatively non-toxic acid or base.

The term "pharmaceutically acceptable" means exhibiting properties that are not toxic to cells or humans exposed to the composition.

The term "pharmaceutically acceptable salt" refers to a salt prepared using a specific compound according to one aspect and a relatively non-toxic acid or base. When the compounds contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include salts of sodium, potassium, calcium, ammonium, organic amines, or magnesium or similar salts. When the compounds contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of an acid in neat solution or in a suitable inert solvent. Pharmaceutically acceptable acid addition salts include salts of inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, hydrogen carbonate ion, phosphoric acid, hydrogen phosphate ion, dihydrogen phosphate ion, sulfuric acid, hydrogen sulfate ion, hydroiodic acid or phosphorous acid; and salts of organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, and methanesulfonic acid. and salts of amino acids (eg, arginine) and salts of organic acids such as glucuronic acid.

The pharmaceutically acceptable salt can be synthesized from a parent compound containing an acidic or basic moiety by conventional chemical methods. Generally, these salts are prepared by reacting the free acid or base form of these compounds with an appropriate stoichiometric amount of the base or acid, either in water or in an organic solvent or in a mixture of the two. In general, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred.

The compound of Formula 1 or a pharmaceutically acceptable salt thereof can activate TGF-β receptors in immature or mature dendritic cells.

Another aspect provides a composition for activating a TGF-β receptor comprising a compound of Formula 2 or a pharmaceutically acceptable salt thereof:

[Formula 2]

Wherein R ₁ to R ₃ and R ₃ ′ are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, and substituted or unsubstituted alkynyl.

"Alkyl", "alkenyl", "alkynyl", "substitution", "salt" and the like may be within the foregoing ranges.

The compound of Formula 2 or a pharmaceutically acceptable salt thereof can also activate TGF-β receptors in immature or mature dendritic cells.

Another aspect provides a composition for activating a TGF-β receptor comprising a compound of Formula 3 or a pharmaceutically acceptable salt thereof:

[Formula 3]

Wherein R ₁ to R ₃ and R ₃ ′ are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, and substituted or unsubstituted alkynyl.

"Alkyl", "alkenyl", "alkynyl", "substitution", "salt" and the like may be within the foregoing ranges.

The compound of Formula 3 or a pharmaceutically acceptable salt thereof can also activate TGF-β receptors in immature or mature dendritic cells.

Another aspect provides a composition for activating a TGF-β receptor comprising a compound of Formula 4 or a pharmaceutically acceptable salt thereof:

[Formula 4]

.

.

"Salt", "pharmaceutically acceptable salt" and the like may be within the foregoing range.

The compound of Formula 4 or a pharmaceutically acceptable salt thereof can also activate TGF-β receptors in immature or mature dendritic cells.

Another aspect includes culturing immature dendritic cells and the composition for activating the TGF-β receptor together; and

Provided is a method for producing dendritic cells with enhanced immune tolerance comprising culturing the cultured immature dendritic cells together with an antigen or a fragment thereof.

The dendritic cell (DC) is an antigen-presenting cell of the mammalian immune system, and is a cell that processes an antigen material, expresses it on the cell surface, and presents it to T cells. The dendritic cells are derived from bone marrow hematopoietic stem cells, and these progenitor cells initially transform into immature dendritic cells, which engulf pathogens and break down their proteins into small peptide fragments, and as they mature, use MHC proteins to Displaying this piece on the cell surface simultaneously activates T cells and B cells.

The immature dendritic cells can be transformed into mature dendritic cells by culturing with antigens or fragments thereof. By culturing the immature dendritic cells and the composition for activating the TGF-β receptor together before culturing with the antigen or fragment thereof, the TGF-β receptor of the immature dendritic cells is activated and the cultured immature dendritic cells are Dendritic cells with enhanced immune tolerance can be prepared by culturing with an antigen or a fragment thereof.

The term "fragment" refers to a part of a substance, and specifically, "the antigen or a fragment thereof" refers to the antigen and a part of the antigen (eg, some amino acids, some sugar chains, a substance in which some amino acids and some sugar chains are linked, etc. ) means.

In one aspect, the antigen may be an autologous antigen.

The term "autoantigen" refers to a tissue or component of one's own that stimulates the formation of autoantibodies, and refers to a tissue or component capable of inducing autoimmunity. Specifically, the antigen may be an antigen capable of inducing autoimmunity, and more specifically, may be collagen.

In the production method according to one aspect, when the antigen is collagen, dendritic cells with enhanced immune tolerance to autoimmune diseases that can be induced by collagen, for example, rheumatoid arthritis, can be prepared.

Another aspect provides a dendritic cell with enhanced immune tolerance prepared according to the above production method.

The term "immune tolerance" refers to a multi-step protection mechanism that prevents immune cells or antibodies from attacking the host, i.e. itself, and the term "immune tolerance" refers to the protective ability to prevent immune cells or antibodies from attacking the host, i.e. itself. it means.

In one aspect, the dendritic cells with enhanced immune tolerance may have reduced expression of at least one cell surface molecule selected from the group consisting of CD80, CD86 and MHC class II, specifically CD80, CD86 and MHC. It may be that the expression of all of class II is reduced.

The dendritic cells with enhanced immune tolerance are cultured with the composition for activating the TGF-β receptor in an immature dendritic cell state, thereby increasing the expression of at least one cell surface molecule selected from the group consisting of CD80, CD86 and MHC class II. may have been reduced.

Further, in one aspect, the dendritic cells with enhanced immune tolerance have reduced expression of at least one inflammatory cytokine selected from the group consisting of IL-12p40, IL-6, IL-1β and TNF-α. Specifically, the expression of all of IL-12p40, IL-6, IL-1β, and TNF-α may be decreased.

Dendritic cells with enhanced immune tolerance are cultured with the composition for activating the TGF-β receptor in an immature dendritic cell state, thereby at least selected from the group consisting of IL-12p40, IL-6, IL-1β and TNF-α. It may be that expression of one inflammatory cytokine is reduced.

In one aspect, the dendritic cells with enhanced immune tolerance are at least one selected from the group consisting of T cell Th1 (CD4 ⁺ IFN-γ ⁺ ) cells and Th17 (CD4 ⁺ IL-17 ⁺ ) cells. It may inhibit differentiation into cells, and specifically, the dendritic cells may inhibit the differentiation of T cells into both Th1 (CD4 ⁺ IFN-γ ⁺ ) cells and Th17 (CD4 ⁺ IL-17 ⁺ ) cells. can

Dendritic cells with enhanced immune tolerance are cultured with the composition for activating the TGF-β receptor in an immature dendritic cell state, thereby increasing T cell Th1 (CD4 ⁺ IFN-γ ⁺ ) cells and Th17 (CD4 ⁺ IL-17 ⁺ ) may inhibit differentiation into at least one cell selected from the group consisting of cells.

Also, in one aspect, the dendritic cells with enhanced immune tolerance may induce differentiation of T cells into Treg (CD4 ⁺ Foxp3 ⁺ ) cells.

Dendritic cells with enhanced immune tolerance may be cultured with the composition for activating the TGF-β receptor in an immature dendritic cell state, thereby inducing differentiation of T cells into Treg (CD4 ⁺ Foxp3 ⁺ ) cells.

Also, in one aspect, the dendritic cells with enhanced immune tolerance may have increased phosphorylation of TGF-β receptor I (TGFBRI).

Dendritic cells with enhanced immune tolerance may be cultured with the composition for activating the TGF-β receptor in an immature dendritic cell state, thereby increasing phosphorylation of the TGF-β receptor (TGF-β receptor I, TGFBRI).

Another aspect provides a pharmaceutical composition for preventing or treating autoimmune diseases comprising the dendritic cells having enhanced immune tolerance.

The "immune tolerance", "dendritic cells with enhanced immune tolerance", etc. may be within the aforementioned range.

As used herein, the term "autoimmune disease" is caused by a process in which a problem occurs in inducing or maintaining self-tolerance, resulting in an immune response against self-antigens, resulting in a phenomenon that attacks one's own tissue. means disease. The self-tolerance refers to immunologic unresponsiveness that does not adversely react to antigenic substances constituting self.

The autoimmune diseases include, for example, multiple sclerosis, inflammatory bowel disease, graft-versus-host disease, asthma, atopy, psoriasis, rheumatoid arthritis, systemic lupus erythematosus, type 1 diabetes, polymyalgia rheumatoid, ankylosing spondylitis, psoriatic arthritis, Eczema, scleroderma, IL-17-induced dementia, peripheral neuritis, uveitis, polymyositis/dermatomyositis, autoimmune cytopenia, autoimmune myocarditis, acute myocardial infarction due to autoimmune myocarditis, atopic dermatitis, primary cirrhosis, dry eye , Fibromyalgia, Goodpeicher syndrome, autoimmune meningitis, Sjogren's syndrome, Addison's disease, alopecia areata, autoimmune hepatitis, autoimmune mumps, dystrophic epidermolysis bullosa, epididymitis, glomerulonephritis, Graves' disease, celiac disease, Guillain-Barre Syndrome, Hashimoto's disease, hemolytic anemia, myasthenia gravis, amyotrophic lateral sclerosis, rheumatoid fever, sarcoidosis, spondyloarthrosis, thyroiditis, vasculitis, vitiligo, myxedema, pernicious anemia, antiphospholipid syndrome, late and chronic rejection of solid organ transplantation, autoimmunity Arthritis, Crohn's disease, autoimmune encephalomyelitis, thyrotoxicosis, autoimmune atrophic gastritis, early menopause, juvenile diabetes, pemphigus, sympathetic ophthalmitis, idiopathic leukopenia, primary biliary sclerosis, chronic active hepatitis, cirrhosis, ulcerative colitis, scleroderma, Wegener Granulomatosis, discoid lupus, systemic sclerosis, mixed connective tissue disease, idiopathic thrombocytopenic purpura, irritable bowel syndrome, narcolepsy, IgA deficiency, systemic lupus erythematosus, gluten-sensitive enteropathy, autoimmune neutropenia, autoimmune infertility and hyperdendrosis It may be at least one autoimmune disease selected from the group consisting of depositional diseases, specifically at least one autoimmune disease selected from the group consisting of rheumatoid arthritis, type 1 diabetes and psoriasis, and more specifically It could be rheumatoid arthritis.

In one aspect, the autoimmune disease may be caused by a self antigen, and the self antigen may be, for example, collagen.

The term "prevention" may refer to any action that suppresses or delays the onset of an autoimmune disease in a subject by administration of a pharmaceutical composition according to one aspect.

The term "treatment" may refer to any activity that improves or beneficially changes the symptoms of an autoimmune disease of a subject by administration of a pharmaceutical composition according to one aspect.

According to one aspect, the dendritic cells with enhanced immune tolerance can exhibit an effect of preventing or treating autoimmune diseases through in vivo immune suppression, and specifically, the dendritic cells with enhanced immune tolerance are TGF-β receptors (TGF-β receptor I, TGFBRI) phosphorylation is increased, the expression of at least one cell surface molecule selected from the group consisting of CD80, CD86 and MHC class II is decreased, IL-12p40, IL-6, IL- A group consisting of Th1 (CD4 ⁺ IFN-γ ⁺ ) cells and Th17 (CD4 ⁺ IL-17 ⁺ ) cells of T cells by reducing the expression of at least one inflammatory cytokine selected from the group consisting of 1β and TNF-α Inhibits differentiation into at least one cell selected from, and induces differentiation of T cells into Treg (CD4 ⁺ Foxp3 ⁺ ) cells, thereby exhibiting an effect of preventing or treating autoimmune diseases.

In addition, in one aspect, the pharmaceutical composition may further include a pharmaceutical composition for preventing or treating other autoimmune diseases in addition to the dendritic cells with enhanced immune tolerance.

That is, the pharmaceutical composition may be provided in combination with other conventionally known compositions for preventing or treating autoimmune diseases or newly developed compositions for preventing or treating other autoimmune diseases.

When the pharmaceutical composition includes a composition for preventing or treating other autoimmune diseases, it is important to mix the composition in an amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by a person skilled in the art.

When the pharmaceutical composition further includes a composition for preventing or treating other autoimmune diseases, the effect of preventing or treating autoimmune diseases becomes more remarkable than when the pharmaceutical composition contains only dendritic cells with enhanced immune tolerance as an active ingredient. A synergistic effect may emerge.

Also, in one aspect, the pharmaceutical composition may be administered alone or in combination with other compositions for preventing or treating autoimmune diseases.

The composition for preventing or treating other autoimmune diseases may be conventionally known compositions for preventing or treating other autoimmune diseases or newly developed compositions for preventing or treating other autoimmune diseases.

The pharmaceutical composition may be administered in parallel with other compositions for preventing or treating autoimmune diseases, simultaneously, separately, or sequentially, and may be administered single or multiple times. Considering all of the above factors, it is important to administer an amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.

When the pharmaceutical composition is administered in combination with other compositions for preventing or treating autoimmune diseases, a synergistic effect in which the effect of preventing or treating autoimmune diseases is more remarkable than when the pharmaceutical composition is administered alone may occur.

The term "administration" refers to introducing a predetermined substance into an individual by an appropriate method, and "subject" refers to all organisms such as rats, mice, livestock, and the like, including humans, which may have autoimmune diseases. As a specific example, it may be mammals including humans.

In one aspect, the route of administration of the pharmaceutical composition includes, but is not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, Topical, sublingual or rectal administration is included, and specifically, the pharmaceutical composition may be administered subcutaneously.

The pharmaceutical composition is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the patient's age, sex, condition, body weight, and active ingredient in the body. Factors including absorption, inactivation rate, excretion rate, patient's disease type, severity, drug activity, drug sensitivity, administration time, route of administration and excretion rate, duration of treatment, severity of obesity, drugs used concurrently And it may be determined according to other factors well known in the medical field, for example, the pharmaceutical composition is 1 x 10 ² cells / μl to 1 x 10 ⁴ cells / μl, 1 x 10 ² cells / μl to 5 x 10 ³ cells/μl, 1 x 10 ² cells/μl to 3 x 10 ³ cells/μl, 5 x 10 ² cells/μl to 1 x 10 ⁴ cells/μl, 5 x 10 ² cells/μl to 5 x 10 ³ cells /μl, 5 x 10 ² cells/μl to 3 x 10 ³ cells/μl, 1 x 10 ³ cells/μl to 1 x 10 ⁴ cells/μl, 1 x 10 ³ cells/μl to 5 x 10 ³ cells/μl Alternatively, it may be administered at a concentration of 1 x 10 ³ cells/μl to 3 x 10 ³ cells/μl.

The pharmaceutical composition may be provided as a pharmaceutical composition including the active ingredient alone or including one or more pharmaceutically acceptable carriers, excipients or diluents.

Specifically, the carrier may be, for example, a colloidal suspension, powder, saline solution, lipid, liposome, microspheres or nano-spherical particles. They may be complexed with or associated with the delivery vehicle and are known in the art such as lipids, liposomes, microparticles, gold, nanoparticles, polymers, condensation reagents, polysaccharides, polyamino acids, dendrimers, saponins, adsorption enhancing substances or fatty acids. It can be delivered in vivo using known delivery systems.

When the pharmaceutical composition is formulated, it is prepared using diluents or excipients such as commonly used lubricants, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, fillers, extenders, binders, wetting agents, disintegrants, surfactants, etc. can Solid preparations for oral administration may include tablets, pills, powders, granules, capsules, etc., and such solid preparations may contain at least one excipient in the composition, for example, starch, calcium carbonate, sucrose ) or by mixing lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral use include suspensions, solutions for oral use, emulsions, syrups, etc., and various excipients such as wetting agents, sweeteners, aromatics, preservatives, etc. may be included in addition to water and liquid paraffin, which are commonly used simple diluents. . Formulations for parenteral administration may include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspensions. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycero-geratin, etc. may be used, and when prepared in the form of eye drops, known diluents or excipients may be used. have.

Another aspect provides a method for preventing or treating an autoimmune disease comprising administering the dendritic cells with enhanced immune tolerance to a subject in need thereof.

The "immune tolerance", "dendritic cells with enhanced immune tolerance", "subject", "administration", "autoimmune disease", "prevention", "treatment" and the like may be within the aforementioned range.

Another aspect is a health functional food for preventing or improving autoimmune diseases containing dendritic cells with enhanced immune tolerance,

The pharmaceutically acceptable salt provides a food-acceptable salt, a health functional food.

The "immune tolerance", "dendritic cells with enhanced immune tolerance", "autoimmune disease", "prevention" and the like may be within the aforementioned range.

The term "improvement" can mean any action that at least reduces the severity of a parameter associated with the condition being treated, eg, a symptom. In this case, the health functional food may be used before or after the onset of the disease, simultaneously with or separately from a drug for treatment, to prevent or improve the autoimmune disease.

The term “acceptable food” means exhibiting properties that are not toxic to cells or humans exposed to the compound.

The term "food chemically acceptable salt" means a salt prepared using a specific compound according to one aspect and a relatively non-toxic acid or base.

In one aspect, the health functional food may further include a health functional food for preventing or improving other autoimmune diseases.

The health functional food may be provided in combination with other known health functional food for preventing or improving autoimmune diseases or newly developed health functional food for preventing or improving other autoimmune diseases.

When the health functional food includes a health functional food for preventing or improving other autoimmune diseases, it is important to mix the amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art. have.

When the health functional food further includes a health functional food for preventing or improving other autoimmune diseases, the effect of preventing or improving autoimmune diseases is higher than when the functional food contains only dendritic cells with enhanced immune tolerance as an active ingredient. Significant synergistic effects can occur.

The health functional food may be consumed in parallel with other conventionally known health functional foods for preventing or improving autoimmune diseases or newly developed health functional foods for preventing or improving other autoimmune diseases, simultaneously, separately, or sequentially. It can be taken as a single or multiple intake. Taking all of the above factors into consideration, it is important to consume an amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.

When the health functional food is consumed in combination with other health functional foods for preventing or improving other autoimmune diseases, compared to when the health functional food containing only dendritic cells with enhanced immune tolerance as an active ingredient is consumed alone, autoimmune disease A synergistic effect in which the prevention or improvement effect of

In the health functional food, the active ingredient may be added to food as it is or used together with other food or food ingredients, and may be appropriately used according to conventional methods. The mixing amount of the active ingredient can be suitably determined depending on the purpose of its use (for prevention or improvement). In general, when preparing food or beverage, the health functional food may be added in an amount of about 15% by weight or less, more specifically about 10% by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be less than the above range.

The health functional food may be formulated as one selected from the group consisting of tablets, pills, powders, granules, powders, capsules and liquid formulations by further including one or more of carriers, diluents, excipients and additives. Foods to which dendritic cells with enhanced immune tolerance according to one aspect can be added include various foods, powders, granules, tablets, capsules, syrups, beverages, gums, teas, vitamin complexes, and health functional foods.

Specific examples of the carrier, excipient, diluent, and additive include lactose, dextrose, sucrose, sorbitol, mannitol, erythritol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium phosphate, calcium silicate, and microcrystalline cellulose. , polyvinylpyrrolidone, cellulose, polyvinylpyrrolidone, methylcellulose, water, sugar syrup, methylcellulose, methyl hydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate and mineral oil It may be at least one selected from.

In addition to containing the active ingredient, the health functional food may contain other ingredients as essential ingredients without particular limitation. For example, it may contain various flavoring agents or natural carbohydrates as additional ingredients like a normal beverage. Examples of the aforementioned natural carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sucrose 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. As flavoring agents other than those mentioned above, natural flavoring agents (thaumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can advantageously be used. can The ratio of the natural carbohydrates may be appropriately determined by a person skilled in the art.

In addition to the above, the health functional food according to one aspect is various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and salts thereof , alginic acid and its salts, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, and the like. These components can be used independently or in combination, and the ratio of these additives can also be appropriately selected by those skilled in the art.

According to one aspect, the newly synthesized compound activates the TGF-β receptor of dendritic cells, thereby enhancing the immune tolerance of dendritic cells. Furthermore, the dendritic cells with enhanced immune tolerance can exhibit an effect of preventing and/or treating autoimmune diseases through an immunosuppressive effect in vivo.

Figure 1 is a diagram confirming the change in the surface antigen molecule expression of dendritic cells according to the treatment of T74.
Figure 2 is a diagram confirming the change in inflammatory cytokine expression in dendritic cells according to the treatment of T74.
Figure 3 is a diagram confirming subtypes of helper T cells after co-culture of T74-treated dendritic cells and T cells.
Figure 4 is a diagram confirming the secretion of cytokines after co-culture of T74-treated dendritic cells and T cells.
5 is a diagram confirming the degree of phosphorylation of TGF-β receptors in dendritic cells according to T74 treatment.
6 is a diagram showing the results of administering T74-treated dendritic cells to a rheumatoid arthritis mouse model.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are intended to illustrate the present invention by way of example, and the scope of the present invention is not limited to these examples.

manufacturing example

Preparation Example 1. Synthesis of (E)-1-(3-Aminophenyl)-3-(2,5-dimethoxyphenyl)prop-2-en-1-one (T74) as a novel synthetic TGF-β agonist

2,5-dimethoxy benzaldehyde (0.62 g, 3.70 mmol) was added to an ethanol mixture (30 mL) in which 3-aminoacetophenone (0.50 g, 3.70 mmol) and NaOH (0.44 g, 11.10 mmol) were dissolved in a 100 mL round bottom flask. Stir at room temperature (17 h). Water was added to the reaction mixture, extracted with ethyl acetate, and the organic layer was collected and washed successively with brine and water. The organic layer was dried with anhydrous MgSO ₄ , filtered, and distilled under reduced pressure to remove the organic solvent. The remaining residue was purified by silica gel column chromatography (eluent: ethyl acetate: hexane = 1:2 → acetate: dichloromethane: hexane = 1:1:1 → methanol: dichloromethane = 1:4) to obtain an orange liquid compound (0.41 g, 39.2%) was obtained.

The prepared (E)-1-(3-Aminophenyl)-3-(2,5-dimethoxyphenyl)prop-2-en-1-one (T74) was the same as the compound of Formula 4:

[Formula 4]

(R _f 0.51 (ethyl acetate : hexane = 1:1); HPLC R _T 4.61 min (purity: 98.0%); ¹ H-NMR (DMSO- d ₆ , 400 MHz) δ 3.79 (s, 3H), 3.84 ( s, 3H), 5.35 (s, 2H), 6.84 (ddd, J = 8.0, 2.0, 1.2 Hz, 1H), 7.02 (dd, J = 9.2, 2.8 Hz, 1H), 7.05 (d, J = 8.8 Hz) , 1H), 7.20 (dd, J = 8.0, 8.0 Hz, 1H), 7.24 (dd, J = 2.0, 1.6 Hz, 1H), 7.30 (ddd, J = 8.0, 1.6, 0.8 Hz, 1H), 7.47 ( d, J = 2.4 Hz, 1H), 7.76 (d, J = 15.6 Hz, 1H), 7.94 (d, J = 15.6 Hz, 1H)).

Preparation Example 2. Preparation of T74-treated dendritic cells

6-week-old C57BL/6 mice (Orient Bio) were purchased and bone marrow cells were collected from the hind limb bones. 10% heat-treated fetal bovine serum (FBS) (Gibco), 1X antibiotic-antimycotic (Gibco), 1X 2-Mercaptoethanol (Gibco), 1X Glutamax (Gibco), 20 ng/mL granulocytes Prepare RPMI medium containing macrophage colony stimulating factor (GM-CSF) (JW creagene) and IL-4 (JW creagene), and inoculate 2 x 10 ⁶ cells per 10 mL of medium in a 100 mm diameter culture dish at 37°C and It was cultured under 5% CO ₂ conditions.

After 3 days, a fresh medium was prepared in which T74 dissolved in DMSO was added to the medium in the culture dish at a concentration of 10 μM, and 10 mL was added to the culture dish. Replace mL with fresh medium. The final concentration of DMSO in the prepared medium was 0.2%.

On day 8, cultured cells were obtained, and 10 μM of T74, 1 μg/mL of lipopolysaccharide (Sigma-Aldrich), and 50 μg of chicken-derived type II collagen (Sigma-Aldrich) were added. Aldrich) for 4 hours at 37° C. and 5% CO ₂ to mature and differentiate immature dendritic cells.

Example

Example 1. Confirmation of immunotolerant characteristics of T74 in prepared dendritic cells

(1) Confirmation of changes in the expression of surface antigen molecules

Antibodies (anti-CD11c-APC antibody, anti-CD80-PE antibody, anti-CD86-FITC antibody, anti-MHC II-PE antibody) labeled with fluorescent substances for cell surface molecules in dendritic cells prepared in Preparation Example 2 and reacted at 4° C. for 20 minutes to stain the cells. Then, the cells were washed with phosphate-buffered saline (PBS), and the washed cells were suspended in PBS and analyzed with a flow cytometer, NovoCyte ^® 3000 (ACEA). The fluorescence intensity of a total of 30,000 cells was measured and analyzed with BD Biosciences' FlowJo10 software.

Specifically, on the 3rd, 6th and 8th days of culture, T74 was treated at a concentration of 10 μM, and on the 8th day, LPS and type II collagen were treated for 4 hours to confirm the expression of cell surface molecules in dendritic cells induced to mature. To do this, a fluorescently labeled antibody was stained and flow cytometry was performed.

As a result, as shown in FIG. 1, it was confirmed that the expression of CD80, CD86, and MHC class II, which are costimulatory molecules of dendritic cells, decreased when treated with T74 compared to NT (Non-treated, control group not treated with T74). The above molecules are required in the process of inducing immune activity, such as the presentation and activation of antigens by dendritic cells to T cells, and their expression increases as the maturity of dendritic cells increases.

Therefore, it was confirmed that dendritic cells prepared by treatment with T74 showed immunotolerant properties.

(2) Confirmation of changes in cytokine secretion

The supernatant was obtained by centrifuging the dendritic cell culture medium and the dendritic cell-T cell co-culture medium, and ELISA was performed to measure the secretion of each cytokine. Interleukin (IL)-12p40 (BD Bioscience), IL-1β (Biolegend), IL-6 (R&D systems), tumor necrosis factor alpha (TNF)-α, interferon (IFN)-γ (Biolegend), IL-17A (Biolegend) and transforming growth factor (TGF)-β ELISA kits were used. Anti-IL-12p40 antibody, anti-IL-6 antibody, anti-IL-1β antibody, anti-TNF-α antibody, anti-IFN-γ antibody, anti-IL-17A antibody, anti-TGF- The β antibody was coated overnight at 4° C., and the culture medium was reacted for 2 hours, followed by reaction with biotin-labeled secondary antibody and avidin-labeled horseradish peroxidase (HRP) for 1 hour. After treatment with TMB (Tetramethylbenzidine) substrate, absorbance was measured at 450 nm (**: p<0.01, ***: p<0.001, ****: p<0.0001).

Specifically, on the 3rd, 6th, and 8th days of culture, T74 was treated at a concentration of 10 μM, and on the 8th day, LPS and type II collagen were treated for 4 hours to obtain a culture medium of dendritic cells inducing maturation, thereby increasing the inflammatory cytokines. The secreted amounts of IL-12p40, IL-6, IL-1β, and TNF-α were measured by ELISA.

As a result, as shown in FIG. 2, it was confirmed that the expression of each inflammatory cytokine decreased when treated with T74 compared to NT (Non-treated, control group not treated with T74).

Therefore, it was confirmed that dendritic cells prepared by treatment with T74 showed immunotolerant properties.

Example 2. Confirmation of function of T74 in prepared dendritic cells

(1) Generation of helper T cells by dendritic cells

For evaluation of T cell differentiation capacity of dendritic cells, dendritic cells prepared in Preparation Example 2 and CD3 ⁺ T cells isolated from C57BL/6 mouse spleen were prepared at a ratio of 1:10 at 37° C. and 5% CO ₂ for about 72 hours. co-cultured. Isolation of T cells was performed by first crushing the spleen, removing red blood cells using ACK lysis buffer (LONZA), and then passing through a nylon wool (Polysciences Inc., Warrington, PA, USA) column. After co-culture, fluorescently labeled anti-CD4-APC antibody (Biolegend), anti-CD25-FITC antibody (BD Antibodies), anti-Foxp3-PE antibody (eBioscience), anti-IFN-γ-PE antibody (BD Antibodies) , Flow cytometry was performed using an anti-IL-17A-PE antibody (BD Antibodies). The ratio of T cell subtypes secreting each cytokine was measured and analyzed with FlowJo10 software.

Specifically, on the 3rd, 6th and 8th days of culture, T74 was treated at a concentration of 10 μM, and on the 8th day, LPS and type II collagen were treated for 4 hours to obtain maturation-induced dendritic cells and isolated from mouse spleens. It was co-cultured with CD4 ⁺ T cells at a ratio of 1:10. After 3 days, T cells were obtained and subtypes of helper T cells were confirmed through flow cytometry.

As a result, as shown in Figure 3, the ratio of Th1 (CD4 ⁺ IFN-γ ⁺ ) cells and Th17 (CD4 ⁺ IL-17 ⁺ ) cells, which play a major role in causing an excessive immune response in autoimmune diseases when treated with T74 decreased. On the other hand, the ratio of Treg (CD4 ⁺ Foxp3 ⁺ ) cells suppressing inflammatory immune cells increased. In addition, as a result of confirming the secretion of cytokines using the co-culture medium, as shown in FIG. 4, it was confirmed that IFN-γ secreted by Th1 cells and IL-17A secreted by Th17 cells significantly decreased. In the case of TGF-β, there was no significant change in the amount of secretion.

Therefore, it was confirmed that dendritic cells prepared by treatment with T74 showed immunotolerant properties.

receptor I, TGFBRI)의 인산화 확인 (2) TGF-β receptor (TGF-

Confirmation of phosphorylation of receptor I, TGFBRI)

Protein extraction solution (PRO-PREP™ Protein Extraction Solution) was added to the dendritic cells prepared in Preparation Example 2 to extract proteins and Western blotting was performed.

After SDS-PAGE was performed on a 7% polyacrylamide gel, the protein was transferred to a PVDF membrane (Millipore), and anti-TGF-β receptor I (TGFBRI) antibody and anti-phospho TGFBRI antibody (Thermo Fisher Scientific), anti-GAPDH (Bioss Antibodies) antibody was reacted overnight at 4°C. Then, HRP-conjugated anti-rabbit IgG antibody and anti-mouse IgG antibody (Santa Cruz Biotechnology) were reacted at room temperature for 2 hours. Bands were detected by processing the ECL substrate (Thermo Fisher Scientific) and quantified by ImageJ software.

Specifically, on the 3rd, 6th, and 8th days of culture, T74 was treated at a concentration of 10 μM, and on the 8th day, LPS and type II collagen were treated for 4 hours to obtain maturation-induced dendritic cells, protein extraction and western Blotting was performed. After SDS-PAGE, total protein and phosphorylated forms of TGFBRI (TGF-β receptor I) were confirmed using anti-TGFBRI antibody, anti-phospho(p) TGFBRI antibody, and anti-GAPDH antibody.

As a result, as shown in Figure 5, it was confirmed that the expression of p-TGFBRI increased.

Through this, it was confirmed that T74 is an agonist of the TGF-β receptor, and that the immunotolerant characteristics and functions of dendritic cells prepared by treating T74 were expressed by signal transduction via the TGF-β receptor.

3. Confirmation of immunosuppressive effect of prepared dendritic cells in vivo

In order to confirm the in vivo immunosuppressive effect of the dendritic cells prepared in Preparation Example 2, the therapeutic effect was confirmed in a rheumatoid arthritis mouse model. Pathological evaluation was performed when dendritic cells prepared by inducing a collagen induced arthritis (CIA) model, which is a representative mouse model of rheumatoid arthritis, were administered twice. After purchasing 6-week-old male DBA/1 mice (Orient Bio) and undergoing a 2-week acclimatization period, 200 μg of type 2 collagen per head was mixed with the same amount of Complete Freund's adjuvant (Sigma Aidrich) and injected subcutaneously into the tail. injected. After 3 weeks, 200 μg of type 2 collagen was mixed with the same amount of Incomplete Freund's adjuvant (Sigma Aidrich) and injected in the same manner, and the prepared dendritic cells were injected subcutaneously into the lower abdomen at 2 x 10 ³ cells/μl per mouse. . Eight days later, dendritic cells were injected again in the same manner. Negative control (NC) without CIA induction, positive control (PC) injected with PBS after CIA induction, experimental group injected with T74-untreated dendritic cells, and dendritic cells prepared by treatment with T74 A total of 4 groups were set as the experimental group injected with , and 4 animals were used in the control group and 10 animals were used in the experimental group respectively. Swelling and deformation of the feet of the mice were observed every 2 to 3 days up to 50 days after the induction of arthritis, and the thickness of the soles of the feet was measured to evaluate the incidence and severity of arthritis. The evaluation criteria are 0 points based on the non-inflamed feet of the negative control group, 1 point for swelling in 1 or 2 toes, 2 points for edema in 3 or more toes or swollen instep, and 2 points for toes and toes. Swelling in both the instep and ankle was scored as 3 points, and the entire foot was swollen and unable to function or the toe and ankle were deformed as 4 points. did

Specifically, on the 3rd, 6th, and 8th days of culture, 10 μM of T74 was treated, and on the 8th day, LPS and type II collagen were treated for 4 hours to obtain maturation-induced dendritic cells, which were then tested in vivo in a collagen-induced arthritis mouse model. experiment was conducted. Arthritis was induced by injecting type 2 collagen twice at 3-week intervals through tail subcutaneous injection, and the prepared dendritic cells were injected at 2 x 10 ³ cells/μl per mouse on the 21st and 29th days after the first collagen injection. The arthritis improvement effect was evaluated.

As a result of evaluating the severity of arthritis according to the evaluation criteria at intervals of 2 to 3 days after the second collagen injection, as shown in FIG. In the group (T74) to which the prepared dendritic cells were added, less edema was observed in the toes, soles, and ankles. The result of measuring the thickness of the sole also showed a tendency similar to that of the score. Through the actual mouse foot pictures below, it can be confirmed that the positive control group and the NT group showed severe swelling in the entire foot, whereas the T74 group showed swelling of about one toe.

Through this, it was confirmed that the treatment effect of arthritis appeared in vivo in mice in which arthritis was induced by dendritic cells prepared by treatment with T74.

Claims (21)

A composition for activating TGF-β receptors comprising a compound of Formula 1 or a pharmaceutically acceptable salt thereof:
[Formula 1]

Wherein R ₁ to R ₃ are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, and substituted or unsubstituted alkynyl,
wherein n represents the number of OR ₃ , n is an integer selected from 1 to 5;
When n is 2 or more, OR ₃ of 2 or more is the same as or different from each other.
A composition for activating a TGF-β receptor comprising a compound of Formula 2 or a pharmaceutically acceptable salt thereof:
[Formula 2]

Wherein R ₁ to R ₃ and R ₃ ′ are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, and substituted or unsubstituted alkynyl.
A composition for activating a TGF-β receptor comprising a compound of Formula 3 or a pharmaceutically acceptable salt thereof:
[Formula 3]

Wherein R ₁ to R ₃ and R ₃ ′ are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, and substituted or unsubstituted alkynyl.
A composition for activating a TGF-β receptor comprising a compound of Formula 4 or a pharmaceutically acceptable salt thereof:
[Formula 4]

.
culturing immature dendritic cells with the composition for activating the TGF-β receptor according to any one of claims 1 to 4; and
A method for producing dendritic cells with enhanced immune tolerance comprising culturing the cultured immature dendritic cells together with an antigen or a fragment thereof.
The method according to claim 5, wherein the antigen is a self antigen, the method of producing dendritic cells with enhanced immune tolerance.
The method according to claim 5, wherein the antigen is collagen, the method of producing dendritic cells with enhanced immune tolerance.
Dendritic cells with enhanced immune tolerance prepared according to the method of claim 5.
The dendritic cell according to claim 8, wherein the expression of at least one cell surface molecule selected from the group consisting of CD80, CD86 and MHC class II is reduced in the dendritic cell with enhanced immune tolerance.
The method according to claim 8, wherein the dendritic cells with enhanced immune tolerance have reduced expression of at least one inflammatory cytokine selected from the group consisting of IL-12p40, IL-6, IL-1β and TNF-α. dendritic cells.
The method according to claim 8, wherein the dendritic cells with enhanced immune tolerance are at least one cell selected from the group consisting of T cell Th1 (CD4 ⁺ IFN-γ ⁺ ) cells and Th17 (CD4 ⁺ IL-17 ⁺ ) cells. To inhibit the differentiation of, dendritic cells.
The dendritic cell according to claim 8, wherein the dendritic cell with enhanced immune tolerance induces the differentiation of T cells into Treg (CD4 ⁺ Foxp3 ⁺ ) cells.
The method according to claim 8, wherein the dendritic cells with enhanced immune tolerance is TGF-β receptor (TGF-β receptor I, TGFBRI) phosphorylation is increased, dendritic cells.
A pharmaceutical composition for preventing or treating autoimmune diseases comprising the dendritic cells with enhanced immune tolerance of claim 8.
The method according to claim 14, wherein the autoimmune disease is multiple sclerosis, inflammatory bowel disease, graft-versus-host disease, asthma, atopy, psoriasis, rheumatoid arthritis, systemic lupus erythematosus, type 1 diabetes, polymyalgia rheumatoid, ankylosing spondylitis, psoriatic arthritis , eczema, scleroderma, IL-17-induced dementia, peripheral neuritis, uveitis, polymyositis/dermatomyositis, autoimmune cytopenia, autoimmune myocarditis, acute myocardial infarction due to autoimmune myocarditis, atopic dermatitis, primary cirrhosis, eye Dryness, fibromyalgia, Goodpeicher syndrome, autoimmune meningitis, Sjogren's syndrome, Addison's disease, alopecia areata, autoimmune hepatitis, autoimmune mumps, dystrophic epidermolysis bullosa, epididymitis, glomerulonephritis, Graves' disease, celiac disease, Guillain Barré syndrome, Hashimoto's disease, hemolytic anemia, myasthenia gravis, amyotrophic lateral sclerosis, rheumatoid fever, sarcoidosis, spondyloarthrosis, thyroiditis, vasculitis, vitiligo, myxedema, pernicious anemia, antiphospholipid syndrome, late and chronic rejection of solid organ transplantation, autologous Immune arthritis, Crohn's disease, autoimmune encephalomyelitis, thyrotoxicosis, autoimmune atrophic gastritis, early menopause, juvenile diabetes mellitus, pemphigus, sympathetic ophthalmitis, idiopathic leukopenia, primary biliary sclerosis, chronic active hepatitis, cirrhosis, ulcerative colitis, scleroderma, Wegener's granulomatosis, discoid lupus, systemic sclerosis, mixed connective tissue disease, idiopathic thrombocytopenic purpura, irritable bowel syndrome, narcolepsy, IgA deficiency, systemic lupus erythematosus, gluten-sensitive enteropathy, autoimmune neutropenia, autoimmune infertility, and At least one autoimmune disease selected from the group consisting of dense deposits, a pharmaceutical composition.
The pharmaceutical composition of claim 14, wherein the autoimmune disease is at least one autoimmune disease selected from the group consisting of rheumatoid arthritis, type 1 diabetes, and psoriasis.
The pharmaceutical composition according to claim 14, wherein the autoimmune disease is caused by collagen.
The pharmaceutical composition according to claim 14, wherein the pharmaceutical composition is administered subcutaneously.
The method according to claim 14, wherein the pharmaceutical composition is administered at a concentration of 1 x 10 ² cells / μl to 1 x 10 ⁴ cells / μl, the pharmaceutical composition.
A method for preventing or treating an autoimmune disease comprising administering the dendritic cells with enhanced immune tolerance of claim 8 to a subject in need thereof.
As a health functional food for preventing or improving autoimmune diseases containing dendritic cells with enhanced immune tolerance of claim 8,
The pharmaceutically acceptable salt is a food pharmaceutically acceptable salt, health functional food.

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