KR20150033978A - Composition for Suppression of Dendritic Cell Maturation Comprising Tussilagone As Active Ingredient - Google Patents

Abstract

The present invention relates to a novel use of inhibiting activity and maturation of dendritic cell of Tussilagone wherein the Tussilagone efficiently inhibits nitric oxide generated from maturated dendritic cells, expression of molecules like MHC-I, MHC-II, CD40, CD80 and CD86 on cell surfaces, phagocyte from working, cytokine like IL-12, TNF-α, IL-1β, IFN-α and IFN-β from being generated, movement to lymph node, mitogen activated protein kinase (MAPK ) from being activated, and inducement performance of T lymphocyte in lymph node from being activated wherein the Tussilagone has a capacity of inhibiting the maturation and activity of dendritic cells and is accordingly used for developing immunosuppressive drug and autoimmune disease treating drug.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for suppressing the maturation of dendritic cells comprising tylsilagon as an active ingredient,

The present invention relates to a composition for inhibiting maturation or activation of dendritic cells comprising tylsilagon as an active ingredient. The present invention also relates to a composition for treating, preventing or ameliorating an autoimmune disease and an immunosuppressive agent comprising tylsylagon as an active ingredient.

Dendritic cells (DCs) are known to be the most important cells capable of inducing anti-tumor immunity while being known to be the cells capable of most effectively inducing the immune response by T lymphocytes. Dendritic cells strongly induce the activation of T lymphocytes with antigen-specific T cell receptors by acquiring antigens through phagocytosis, treating them in cells, and expressing antigenic peptides on MHC. In addition, activated dendritic cells express cytokines such as IL-12, TNF-α, and IL-1 to inhibit T-lymphocyte apoptosis, induce T lymphocyte differentiation and CTL activity, To increase anti-tumor immunity.

Dendritic cells are known to induce immune tolerance through a variety of mechanisms besides their ability to induce an immune response. Recent studies have shown that persistent maturation and activation of dendritic cells are involved in autoimmune diseases such as systemic lupus erythematosus (Lupus) and Sjogren's syndrome. Thus, inhibiting the maturation of dendritic cells is thought to be useful for the treatment of such autoimmune diseases.

Kantou (Tussilago farfara L.) is a perennial plant of Asteraceae widely distributed throughout the world such as North Asia, Southwest Asia, Europe, North Africa and North America. It is cultivated only medicinally in Korea. It grows using seeds and undergrowth, and the flowering season is from February to March. It blooms bright yellow flowers similar to dandelions and has round, wide leaves. The length of the flowering stems grows up to 5-30 cm and has the characteristic that the flowers bloom before it leaves in early spring. The Kanto region is known to have the efficacy of the lungs to soak dry things, to lower the flies, to stop the coughing, and to stop coughing. The Kanto flower bud called "Kantouhwa" has long been used as a cough suppressant for respiratory diseases such as asthma and bronchitis Has been used. Kanto is known to have effects such as gonadal function, antiinflammatory action, vasopressor action, central nervous stimulation action, vasoconstrictor action, parasympathetic action, bronchial relaxation action, and in oriental medicine, it regulates gonads, sprains, coughs, It is used as a herbal medicine. The Kanto extract contains active ingredients such as chlorogenic acid, rutin, and tetracylone. Of these, tubylagon is reported to inhibit macrophage activation and inflammatory cytokine secretion. However, no studies have been reported on the activity of tubular RNA and dendritic cells.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

Korean Patent Publication No. 10-2007-0008228

The present inventors have sought to discover candidate substances that can be developed as immunosuppressive drugs and autoimmune disease drugs through the activity of inhibiting the activation and maturation of dendritic cells, which are powerful antigen presenting cells, from natural products. As a result, the present inventors have completed the present invention by experimentally proving that Tusilagon extracted from Kanto region effectively inhibits maturation and activation of dendritic cells.

Accordingly, it is an object of the present invention to provide a composition for inhibiting dendritic cell maturation or activation comprising tylsilagon as an active ingredient.

Another object of the present invention is to provide an immunosuppressive agent comprising tylsilagon as an active ingredient.

It is still another object of the present invention to provide a composition for treating, preventing or ameliorating an autoimmune disease comprising tylsilagon as an active ingredient.

The objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, there is provided a composition for inhibiting the maturation or activation of dendritic cells comprising tylsilagon as an active ingredient.

According to another aspect of the present invention, there is provided an immunosuppressive agent comprising a tubylagon as an active ingredient.

According to still another aspect of the present invention, there is provided a composition for treating, preventing or ameliorating an autoimmune disease comprising tylsilagon as an active ingredient.

The present invention is based on experimentally demonstrating that tylsylagon effectively inhibits maturation and activation of immature dendritic cells into mature resin cells.

Tussilagone, an active ingredient of the present invention, is a compound represented by the following formula (1).

The effective ingredient of the composition of the present invention, tucilagon, may be a natural product, for example, extracted from Kanto and may be chemically synthesized. It is apparent to those skilled in the art that chemically synthesized tetracalon has the same activity as that extracted from the natural product.

As used herein, the term "dendritic cell (DC)" is used to absorb an antigen into cells and treat it to present an antigen or antigen-derived peptide with a major histocompatibility complex class I complex or MHC class II complex Which is a specialized antigen presenting cell. The dendritic cells in the present invention are meant to refer to cells having the typical phenotype and characteristics of dendritic cells presented by Steinman et al. (Annual Rev. Immunol. 9: 271-296, 1991 and Banchereau and Steinman Nature 392: 245-252, do. Dendritic cells contain both immunogenic and immuno tolerogenic antigen presenting cells and are classified into immature dendritic cells (semimature DC) and mature dendritic cells (mature dendritic cells) according to their maturity Can be classified.

The term "dendritic cell maturation or activation" as used herein refers to the process by which dendritic cells acquire their capacity as professional antigen presenting cells.

The maturation of dendritic cells is mediated by interactions between molecules derived from pathogens such as bacteria or viruses, such as LPS, CpG DNA, and dsRNA and various types of TLR receptors capable of recognizing them.

Dendritic cells initially determine whether the immune system will trigger an inflammatory response or tolerance to stimulants and then determine which T lymphocytes inexperienced T lymphocytes will differentiate into T lymphocytes It plays a key role in determining. Immature dendritic cells capture antigens by endocytosis through phagocytosis or receptors, process antigens through a series of processes in cells, mount antigenic peptides on MHC and present them to T lymphocytes. Along with the process of antigen processing, dendritic cells become more mature and lose receptors used for phagocytosis and intracellular entry, and the expression of surface molecules such as MHC class I, MHC class II, CD (cluster determinant) 80, CD86 and CD40 , And expresses a new cytokine receptor, which migrates to a region rich in T lymphocytes in the surrounding lymph nodes and presents an antigen to T lymphocytes, resulting in a T lymphocyte immune response.

The tubylagon of the present invention effectively inhibits the maturation and activation of dendritic cells induced by lipopolysaccharide (LPS), as demonstrated in the following specific example. More specifically, the cytosol argon is produced by nitric oxide (NO) -induced production of lipopolysaccharide (LPS) -treated dendritic cells, expression of cell surface molecules such as MHC-I, MHC-II, CD40, CD80 and CD86, , Activation of lymphocytes, activation of mitogen-activated protein kinase (MAPK), and activation of T lymphocytes in the lymph nodes, as well as the production of cytokines such as IL-12, TNF-a, IL-1 ?, IFN- ?, IFN-? Suppress induction.

According to another aspect of the present invention, there is provided an immunosuppressive agent comprising a tubylagon as an active ingredient.

The maturation and activation of dendritic cells are essential for inducing an immune response by T lymphocytes and immature dendritic cells are known to exhibit immunosuppressive activity by inducing immune tolerance. Therefore, in the present invention having dendritic cell maturation and activation inhibitory activity Can be useful as an immunosuppressive agent.

The cytosol argon of the present invention can be used for diseases or conditions requiring immunosuppression and can be useful as an immunosuppressive agent for preventing graft rejection of tissues or organs mediated by, for example, an immune response. The graft rejection refers to the fact that the genetic background between the donor and the recipient of the graft (the cell, tissue or organ as part of the transplanted organism) differs (1) the disease caused by the donor's graft-derived immune cells recognizing the consumer as an external substance (Ii) graft-versus-host disease) and (2) diseases caused by the recipient's attack by recognizing the graft of the donor as an external substance (i.e. graft rejection). Examples of implanted tissues and organs causing rejection include not only the heart, kidney, liver, cartilage, skin, cornea, lung, pancreas, small intestine, limb, muscle, nerve, duodenum, small intestine, pancreatic insulin- , And graft-versus-host disease caused by cartilage transplantation.

According to still another aspect of the present invention, there is provided a composition for treating, preventing or ameliorating an autoimmune disease comprising tylsilagon as an active ingredient. The maturation and activation of dendritic cells are essential for inducing an immune response by T lymphocytes. Immature dendritic cells are known to exhibit immunosuppressive activity by inducing immune tolerance. Therefore, it is known that dulcimer Can be usefully used for the treatment, prevention and improvement of autoimmune diseases.

As used herein, the term " autoimmune disease "refers to a disease caused by the production of antibodies against the body's own tissues, tissues or components. The autoimmune diseases to be treated, prevented or ameliorated in the present invention include, for example, rheumatoid arthritis, multiple sclerosis, type I diabetes mellitus, myasthenia gravis, systemic lupus erythematosus (lupus), Sjogren's syndrome, Graves' But are not limited to, chronic colitis, Crohn's disease, ankylosing spondylitis, Hashimoto's disease, inflammatory bowel disease, experimental autoimmune encephalomyelitis, and neutropenia. More preferably, the autoimmune disease is systemic lupus erythematosus (lupus), or Sjogren's syndrome.

The composition of the present invention may be provided in the form of a pharmaceutical composition. In this case, the pharmaceutical composition of the present invention may contain a pharmaceutically acceptable carrier in addition to the active ingredient tylsylagon. Such carriers are those conventionally used in the field of the art and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone , Cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, . Meanwhile, the oral dose of the pharmaceutical composition of the present invention is preferably 0.001 to 1000 mg / kg (body weight) per day.

The pharmaceutical composition of the present invention can be administered orally or parenterally. When administered parenterally, it can be administered by intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, transdermal administration, and the like.

The concentration of the active ingredient contained in the composition of the present invention can be determined in consideration of the purpose of treatment, the condition of the patient, the period of time required, and the like, and is not limited to a specific range of concentration.

The pharmaceutical composition of the present invention may be formulated into a unit dose form by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by a person having ordinary skill in the art to which the present invention belongs. Or by intrusion into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in an oil or aqueous medium, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

The composition of the present invention may also be provided in the form of a functional food composition for improving autoimmune diseases. In this case, the food composition of the present invention may contain, in addition to the active ingredient, . For example, proteins, carbohydrates, fats, nutrients, seasonings and sweeteners. Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. As a sweetening agent, a natural sweetening agent (e.g., tau Martin, stevia extract (e.g., rebaudioside A, glycyrrhizin)) and a synthetic sweetener (saccharin, aspartame, etc.) may be used. For example, when the food composition of the present invention is prepared as a drink, it may further contain citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, mulberry extract, jujube extract, licorice extract, have.

The present invention relates to new uses for dysplasia cell maturation and inhibition of activation of tubylagon. The present invention provides a method for producing a cytosol of the present invention comprising the steps of: (a) culturing a dorsal root of a tubular epithelial cell; (b) effectively inhibits the production of cytokines such as IL-1, IL-1β, IFN-α and IFN-β, migration into lymph nodes, activation of mitogen-activated protein kinase (MAPK), and activation of T lymphocyte activation in lymph nodes . The tubylagon of the present invention has the activity of inhibiting maturation and activation of dendritic cells, and thus can be developed as an immunosuppressant and an autoimmune disease therapeutic agent.

Figure 1 shows the inhibitory effect of the tetracylagon on the production of nitric oxide (NO) in dendritic cells induced by treatment with lipopolysaccharide.
2 shows the inhibitory effect of the tetracylagon on the change of the phenotype of dendritic cells induced by treatment with lipopolysaccharide.
Fig. 3 shows the inhibitory effect of the tetracylagon on the change of the phagocytic action of dendritic cells induced by treatment with lipopolysaccharide.
Fig. 4 shows the inhibitory effect of the cytosol on the secretion of cytokines of dendritic cells induced by treatment with lipopolysaccharide.
Fig. 5 shows the effect of the tetrazolone inhibiting effect on the migration ability of dendritic cells induced by treatment with lipopolysaccharide.
FIG. 6 shows the effect of the tetracylagon on the change of the signal transduction system of dendritic cells induced by treatment with lipopolysaccharide.
Fig. 7 shows the inhibitory effect of the tylosilone on the T lymphocyte activation ability of dendritic cells induced by treatment with lipopolysaccharide.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

EXAMPLE 1 Influence of the Tylsylagon on Dendritic Cell Maturation Induced by Lipopolysaccharide

Preparation Example 1: Preparation of experimental animals

The experimental animals were fed C57BL / 6 female mice (BioLink, Chungbuk), weighing 18-22g, and the animals were sacrificed at a constant temperature (temperature: 21 ± 2 ℃, The animals were given free access to water and food until the start of the experiment. Before the start of the experiment, the animals were kept in the animal breeding room for 7 days.

Preparation Example 2: Preparation of dendritic cells

Bone marrow cells were isolated from C57BL / 6 mice and cultured at a concentration of 2 × 10 ⁵ cells / ml. GM-CSF was added at a concentration of 2 ng / ml for 8 days, and the dendritic cells were known to be immature dendritic cells.

Experimental Example 1: Effect of dendritic cells on nitric oxide (NO) production

Immature dendritic cells were prepared in the same manner as in Production Example 2. These immature dendritic cells were plated on 96-well plates at a concentration of 5 × 10 ⁵ cells / ml and stabilized for 2 hours. Tucilagon was added to the dendritic cells at a concentration of 1-30 μM, lipopolysaccharide was added at a concentration of 1 μg / ml, and the cells were cultured for 24 hours. After incubation, 100 μl of griess A (g = 1: 1) was added to the supernatant (100 μl) and incubated at room temperature for 10 minutes. Absorbance was measured at 540 nm to quantitate NO production.

As shown in FIG. 1, the production of nitric oxide (NO) was 4% in the control group and 100% in the lipopolysaccharide-treated group. In the experimental group treated with lipopolysaccharide and tetracylagon simultaneously And decreased to 89, 68, 46, and 2% depending on the concentration.

Experimental Example 2: Effect of dendritic cells on expression of cell surface factors

Immature dendritic cells were prepared in the same manner as in Production Example 2. To these immature dendritic cells were added tetracylon at a concentration of 1-30 μM and lipopolysaccharide was added at a concentration of 1 μg / ml and cultured for 24 hours. Cells were collected and stained with antibodies such as FITC (Fluorescein Isothiocyanate) -MHC I, FITC-MHC II, FITC-CD40, FITC-CD80, FITC-CD86, and PE (R-Phycoerythrin) Analysis was performed using a flow cytometer and the results were expressed as Mean Fluorescence Intensity (MFI). The higher the MFI value, the stronger the expression of the surface factor, and also the greater the maturation of the dendritic cells. CD11c is a surface factor selectively expressed in dendritic cells, and MHC-I, MHC-II, CD40, CD80, and CD86 are surface factors that are expressed more as dendritic cells mature.

As shown in FIG. 2, the expression level of MHC-I was 19.88 in the control group, 93.89 in the lipopolysaccharide-treated group, 93.16 in the experimental group treated with lipopolysaccharide and tetrazolone, 85.97, 79.35 and 63.51, respectively. The expression level of MHC-II was decreased to 61.52, 58.46, 52.97, and 34.76, respectively, in the experimental group treated with lipopolysaccharide and tetracylon at a concentration of 18.26 in the control group and 65.51 in the lipopolysaccharide treated group Respectively. The expression level of CD40 was 18.02 in the control group and increased to 99.75 in the lipopolysaccharide treatment group and decreased to 96.64, 93.71, 72.28, and 52.11 in the experimental group treated with lipopolysaccharide and tetrazolone simultaneously. The expression level of CD80 was increased to 35.98 in the control group, 19.11 in the control group, and decreased to 34.66, 34.20, 28.77 and 21.16 in the experimental group treated with lipopolysaccharide and tetracylagon simultaneously. The expression level of CD86 was increased to 80.58 in the control group, 19.24 in the control group, 80.58 in the lipopolysaccharide treatment group, and decreased to 78.72, 75.84, 65.00 and 32.36 in the experimental group treated with lipopolysaccharide and tetrazolone simultaneously.

Experimental Example 3: Effect of dendritic cells on antigen uptake

Immature dendritic cells are good for antigen acquisition, but mature dendritic cells are known to decrease antigen acquisition capacity. It was expected that the action of the lipopolysaccharide would reduce the phagocytosis to a relatively small extent if the tylosilagon inhibited the maturation of the dendritic cells induced by the lipopolysaccharide.

Immature dendritic cells were prepared in the same manner as in Production Example 2. These immature dendritic cells were cultured for 24 hours by adding 1 - 30 μM of tylsylagon and 1 μg / ml of lipopolysaccharide. After culturing, the cells were collected and treated with FITC-dextran at a concentration of 0.7 mg / ml for 2 hours. Cells were washed and the excess FITC-dextran was completely removed and the amount of antigen introduced into dendritic cells was measured using a flow cytometer. Dendritic cells were stained with CD11c-PE antibody selectively binding to dendritic cells to analyze the degree of antigen acquisition after selectively separating and analyzing dendritic cells.

As can be seen from FIG. 3, when the cells were cultured at 37 ° C for 2 hours, the antigen acquisition of the control group was 40%, and the antigen acquisition of lipopolysaccharide-treated dendritic cells was reduced to 19%. In addition, unlike earlier estimates, the antigen uptake was further reduced to 16, 14, 12, and 12% depending on the concentration in the experimental group treated with 1 - 30 μM tetrazolone and 1 μg / ml lipopolysaccharide. This means that the tetracylagon inhibits the ability of the dendritic cells to acquire an antigen, a unique function. When cultured at 4 ° C, the cells were completely stopped from acquiring the antigen, and all the cells showed low 2-4% acquisition of antigen. This means that antigen acquisition is an active immune response.

Experimental Example 4: Effect of dendritic cells on cytokine production

Immature dendritic cells were prepared in the same manner as in Production Example 2. To these immature dendritic cells were added tetracylon at a concentration of 1-30 μM and lipopolysaccharide was added at a concentration of 1 μg / ml and cultured for 24 hours. After cultivation, the cells were harvested and RNA was isolated from dendritic cells. The amount of cytokine expressed by reverse transcription-polymerase chain reaction (RT-PCR) was measured and the amount of cytokine secreted from the dendritic cells And measured by enzyme-linked immunoassay. Materials were purchased from R & D Systems and tested according to the test methods provided by the company.

As can be seen from the panel A of FIG. 4 and the panel B of FIG. 4, the control group hardly secreted cytokines of IL-12, TNF-a, IL-1 ?, IFN- ?, IFN-? The secretion of these cytokines was strongly increased in the rye treatment group, but the secretion of cytokines was inhibited in a concentration dependent manner when the lipopolysaccharide and the tetracylagon were treated at the same time. From these results, it can be seen that the maturation of dendritic cells by lipopolysaccharide is inhibited by the tetracylagon.

Experimental Example 5: Influence on the migration ability of dendritic cells

When dendritic cells capture and mature the antigen, they migrate to lymph nodes (LNs) with T cells to activate T cells. Immature dendritic cells express CC chemokine receptors (CCR) 1 and CCR5, which are chemokine receptors related to inflammation, but the expression of CCR7 increases as maturation progresses. Changes in chemokine receptors on the surface of dendritic cells affect the ability of dendritic cells to migrate. Immature dendritic cells were prepared in the same manner as in Production Example 2. To these immature dendritic cells were added tetracylagon at a concentration of 30 μM and lipopolysaccharide was added at a concentration of 1 μg / ml and cultured for 4 hours. After incubation, the cells were harvested and the expression of chemokine receptor was measured by RT-PCR (reverse transcription-polymerase chain reaction).

As can be seen from panel A of FIG. 5, the CCR1 and CCR5 genes were strongly expressed in the control group and the expression level was decreased in the lipopolysaccharide-treated group. However, the expression levels of CCR1 and CCR5 genes were increased in a concentration-dependent manner in the group treated with both tetracylone and lipopolysaccharide. On the other hand, the CCR7 gene was hardly expressed in the control group, but strongly expressed in the lipopolysaccharide treated group, and the CCR7 gene expression was decreased in the concentration-dependent manner in the group treated with the tetracylagon.

Next, a change in the migration ability of dendritic cells was confirmed using a transwell. Immature dendritic cells were prepared in the same manner as in Production Example 2. To these immature dendritic cells were added tetracylon at a concentration of 30 μM and lipopolysaccharide was added at a concentration of 1 μg / ml and cultured for 24 hours. After incubation, the cells were collected and dendritic cells were dispensed into the upper chamber at a concentration of 2 × 10 ⁶ cells / ml. In the lower chamber, RANTES, a ligand of CCR5, and MIP-3β, a ligand of CCR7, Were added at concentrations of 30-300 ng / ml, respectively, and incubated at 37 ° C for 6 hours. The number of cells transferred to the lower chamber was then measured using a flow cytometer.

As can be seen from the panel B of FIG. 5, the control group did not show any change in the movement due to MIP-3 ?, and the movement increased depending on the concentration of RANTES. In the lipopolysaccharide-treated group, the movement was increased depending on the concentration of MIP-3? As opposed to the control group, and the movement of RANTES did not change. In the group treated with both tylsylagon and lipopolysaccharide, the movement was increased in both MIP-3β and RANTES, but the movement was more dependent on the concentration of RANTES than MIP-3β. From this result, it was found that the change of the migration ability of dendritic cells by lipopolysaccharide was suppressed by the cytochalasin.

Experimental Example 6: Effect of dendritic cells on signal transduction system

Dendritic cells are known to activate mitogen-activated protein kinases such as ERK, JNK, and p38 during maturation. Immature dendritic cells were prepared in the same manner as in Preparation Example 2, and then treated with 1 to 30 μM of tylsylagon and 1 μg / ml of lipopolysaccharide for 20 minutes, and the cells were disrupted to separate the proteins. The amount of phosphorylated ERK, JNK, and p38 protein was measured by a Western blotting method using a specific antibody.

6, the amount of phosphorylated p-JNK and p-p38 increased in the lipopolysaccharide-treated group but decreased in the concurrent treatment group of the lipopolysaccharide and the tylsilagon in a concentration-dependent manner. The amount of p-ERK did not decrease.

Dendritic cells are known to activate NF-κB (nuclear factor-kappa B) during maturation. Immature dendritic cells were prepared in the same manner as in Preparation Example 2, and lipopolysaccharide and tylsylagon were treated for 20 minutes. Cell nuclei were separated and then disrupted to separate all proteins. The amount of NF-κB in the nucleus was confirmed by p65, a subunit of NF-κB, and measured by western blotting.

As shown in panel B of FIG. 6, although the amount of p65 in the nucleus was increased in the lipopolysaccharide-treated group compared to the control group, it was confirmed that the concentration of the tetrazolone was decreased in the group treated with lipopolysaccharide and the tetrazolone simultaneously.

Tucilagon inhibited the migration of NF-κB p65 into the nucleus, and therefore it was tested whether it inhibits IkB degradation at the higher level. Immature dendritic cells were prepared in the same manner as in Preparation Example 2, and then treated with 1 to 30 μM of tylsylagon and 1 μg / ml of lipopolysaccharide for 20 minutes, and the cells were disrupted to separate the proteins. The degradation of IκB-α and IκB-β was measured by a western blotting method.

As shown in panel C of FIG. 6, the amounts of IκB-α and IκB-β were abundant in the control group, but the amount of IκB-α and IκB-β was decreased when the lipopolysaccharide was treated. In the group treated with both lipopolysaccharide and tetracylagon It was confirmed that the decomposition was inhibited depending on the concentration of the tylsilagon.

Example 2: Influence of Tylsylagon on the T lymphocyte Activation Capacity of Dendritic Cells Induced by Lipopolysaccharide

Experimental Example 1: Effect of dendritic cells on T lymphocyte activation activity

The most important immune function of mature dendritic cells is activation of T lymphocytes. Immature dendritic cells acquire antigens, undergo maturation, migrate to immune organs with T cells, and activate T cells. T cells activated by dendritic cells actively proliferate and secrete cytokines.

Immature dendritic cells prepared by the same method as in Preparation Example 2 were treated with 30 μM of tylsylagon and 1 μg / ml of lipopolysaccharide, followed by culturing for 24 hours. After cultivation, the cells were harvested and mixed with allogeneic T cells for 3 days. Cell culture During the last 16 hours, [ ³ H] -thymidine was treated to infiltrate into the DNA of the proliferating T cells, and cell proliferation was measured.

The dendritic cells treated with lipopolysaccharide as in the panel A of FIG. 4 showed strong T lymphocyte proliferative capacity, but it was confirmed that the dendritic cells treated with the lipopolysaccharide and the tetracylagon were inhibited the T lymphocyte proliferative capacity. The amount of IFN-γ and IL-2 secreted by T cells was measured. As can be seen from the panel B of FIG. 4 and the panel C of FIG. 4, the amount of cytokine production was strongly increased in the T lymphocyte co-cultured with dendritic cells treated with lipopolysaccharide, but the lipopolysaccharide- T lymphocytes co-cultured with dendritic cells were inhibited cytokine production. This result implies that the treatment of tubalagon inhibits the dendritic cell maturation by lipopolysaccharide and suppresses T lymphocyte activation by dendritic cells.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (8)

1. A composition for inhibiting maturation or activation of dendritic cells comprising a tubular ligand represented by the following formula (1) as an active ingredient.
[Chemical Formula 1]

Claims 1. An immunosuppressant comprising as an active ingredient a tubylagon represented by the following formula (1).
[Chemical Formula 1]

3. The immunosuppressant according to claim 2, wherein the cytosol argon inhibits maturation or activation of dendritic cells.
A composition for treating, preventing or ameliorating an autoimmune disease, which comprises a tubularagon represented by the following formula (1) as an active ingredient.
[Chemical Formula 1]

5. The method of claim 4, wherein the autoimmune disease is selected from the group consisting of rheumatoid arthritis, multiple sclerosis, Type I or type II diabetes, myasthenia gravis, systemic lupus erythematosus (lupus), Sjogren's syndrome, Graves disease, glomerulonephritis, psoriasis, ulcerative colitis , Crohn's disease, ankylosing spondylitis, Hashimoto's disease, inflammatory bowel disease, experimental autoimmune encephalomyelitis, or immunodeficiency.
5. The composition of claim 4, wherein the tetrazolone inhibits maturation or activation of dendritic cells.
5. The composition of claim 4, wherein said composition is in the form of a pharmaceutical composition for the treatment or prevention of autoimmune diseases.
5. The composition of claim 4, wherein said composition is in the form of a functional food composition for the amelioration of autoimmune disease.

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