KR101317507B1 - Composition for preventing or treating cancer or immune disease comprising rebamipide - Google Patents

Abstract

The present invention relates to a composition for the prevention or treatment of cancer or immune diseases comprising levamipid as an active ingredient, and more specifically, the present invention comprises a levamifeed compound or a pharmaceutically acceptable salt thereof as an active ingredient. It relates to a composition for preventing or treating cancer or immune disease. The levamifeed compound according to the present invention has an excellent effect of inhibiting the differentiation of cytotoxic Th17 cells that produce and secrete inflammatory cytokines, and has the property of inhibiting the function of abnormally activated immune cells and controlling the inflammatory response. Since it has an excellent effect of enhancing the activity of immunoregulatory T cells (Treg), it is a pharmaceutical that can prevent or treat immune diseases such as autoimmune diseases, inflammatory diseases, and transplant rejection diseases caused by various cancer diseases or immune control disorders. It can be usefully used as a composition or an immunosuppressant.

Description

Composition for preventing or treating cancer or immune disease comprising rebamipide as an active ingredient

The present invention relates to a pharmaceutical use of levamipid used to prevent or treat cancer or immune diseases caused by an abnormal immune response, and more particularly, to a cancer or immune disease comprising levamipid as an active ingredient. It relates to a prophylactic or therapeutic composition.

T cells are one of the cell groups that plays a central role in the immune system as a bio-defense system for various pathogens. T cells are produced in the thymus of the human body, and undergo a series of differentiation processes, resulting in differentiation into T cells with inherent characteristics. The differentiated T cells are largely classified into type 1 (Th1) Auxiliary cells (Th2). Among them, the main function of Th1 cells is involved in cell mediated immunity, Th2 cells are involved in humoral immunity, and in the immune system, these two cell populations are balanced with each other so that they are not activated with each other.

Therefore, most of the immune diseases are caused by the imbalance between these two immune cells. For example, when the activity of Th1 cells abnormally increases, an autoimmune disease may occur and the activity of Th2 cells abnormally increases It is known that immune diseases are caused by hypersensitivity reactions.

Meanwhile, according to a recent study on the differentiation of Th1 cells, the existence of a new group of immunoregulatory T cells (Tregs), which can control Th1 cell activity, is known, and studies on the treatment of immune diseases using the same are emerging. In addition, since Treg cells inhibit the function of abnormally activated immune cells to control the inflammatory response, many experiments have been reported to treat immune diseases through the action of increasing the activity of Treg cells.

In addition to the Treg cells, another group made during the differentiation is Th17 cells, which are known to be formed through a process similar to the differentiation of Treg cells during the differentiation of undifferentiated T cells. In other words, differentiation of Treg cells and Th17 cells occurs in the presence of TGF-ß but does not require IL-6 for Treg cells, whereas IL-6 is present with TGF-ß in Th17 cells. Differentiate from In addition, differentiated Th17 cells are characterized by the secretion of IL-17.

Unlike Treg cells, Th17 cells have been found to be involved in the forefront of inflammatory reactions seen in immune diseases, maximizing the signal of inflammatory responses and accelerating disease progression. Therefore, in the case of autoimmune diseases which are not controlled by Treg cells among autoimmune diseases, development of therapeutic agents for autoimmune diseases that target the inhibition of Th17 cell activity has been highlighted.

Currently used immunosuppressive drugs are immunosuppressants that block signal transduction pathways in T cells. These immunosuppressive agents are toxic, infection, lymphoma, diabetes, tremor, headache, diarrhea, high blood pressure, and nausea. There is a problem that side effects such as renal failure occur.

In addition to the treatment of immune diseases through a method of inhibiting the activation of T cells, therapies that regulate the amount of cytokines secreted from immune cells (see Korean Patent Publication No. 2008-0068767) and cytokines secreted from immune cells Therapies using antibodies targeting kine (see Korean Patent Publication No. 2000-0034847 and Korean Patent Publication No. 2002-0020730) are under development. However, this method has to take a long time to apply to the patients after the actual clinical trials, the method using the antibody has a problem that too much cost in the antibody manufacturing process.

Therefore, there is a need for the development of new cancer and immune disease treatment agents that have no side effects and are inexpensive and have excellent therapeutic effects.

Accordingly, the present inventors have confirmed that rebamipide inhibits Th17 activity and increases Treg cell activity, thereby confirming that the rebamipide can be used as a therapeutic agent in various tumors or all immune-related diseases, and completed the present invention.

Accordingly, an object of the present invention is to prevent cancer or immune disease comprising a levami feed compound or a pharmaceutically acceptable salt thereof as an active ingredient to effectively prevent or treat immune diseases caused by various cancers and immune response abnormalities. Or to provide a therapeutic composition.

Another object of the present invention is to reduce or inhibit the differentiation of undifferentiated T cells into Th17 cells in vitro, comprising treating undifferentiated T cells with a levamifeed compound or a pharmaceutically acceptable salt thereof. To provide a way.

Another object of the present invention is a method of activating regulatory T cells comprising treating a LevamiFeid compound or a pharmaceutically acceptable salt thereof to Regulatory T cells (Treg) in vitro. To provide.

Furthermore, another object of the present invention is to provide an immunosuppressive composition comprising a levamifeed compound or a pharmaceutically acceptable salt thereof as an active ingredient.

In order to achieve the above object, the present invention provides a composition for the prophylaxis or treatment of cancer or immune disease comprising a levamifeed compound or a pharmaceutically acceptable salt thereof as an active ingredient.

In one embodiment of the present invention, the levamipid can reduce or inhibit the differentiation of undifferentiated T cells into Th17 cells.

In one embodiment of the present invention, the levamipid may promote or increase the activity or amplification of Regulatory T cells (Treg).

In one embodiment of the present invention, the cancer is colon cancer, rectal cancer, anal muscle cancer, bone cancer, gastric cancer, cerebrospinal tumor, head and neck cancer, thymic tumor, mesothelioma, esophageal cancer, biliary tract cancer, bladder cancer, testicular cancer, small intestine cancer, germ cell tumor, uterus Endocarcinoma, fallopian tube carcinoma, vaginal carcinoma, vulvar carcinoma, multiple myeloma, sarcoma, endocrine adenocarcinoma, thyroid cancer, parathyroid cancer, adrenal cancer, bladder cancer, urethral cancer, pituitary adenoma, renal pelvic carcinoma, spinal cord tumor, multiple myeloma, glioma, central CNS central nervous system tumor, hematopoietic tumor, fibrosarcoma, neuroblastoma, astrocytoma, breast cancer, cervical cancer, ovarian cancer, prostate cancer, pancreatic cancer, kidney cancer, liver cancer, brain cancer, lung cancer, lymphoma, leukemia, malignant melanoma and It can be selected from skin cancers.

In one embodiment of the present invention, the immune disease may be selected from the group consisting of autoimmune diseases, inflammatory diseases and transplantation rejection diseases of cells, tissues or organs.

In one embodiment of the present invention, the immune disorder is selected from the group consisting of Behcet's disease, multiple myelitis / dermatomyositis, autoimmune hemolytic disease, autoimmune myocarditis, atopic dermatitis, asthma, primary cirrhosis, dermatomyositis, Atherosclerosis, neurodegenerative disease, meningitis, Sjogren's syndrome, systemic lupus erythematosus, Addison's disease, alopecia areata, ankylosing spondylitis, autoimmune hepatitis, autoimmune mumps, Crohn's disease, insulin dependent diabetes mellitus, eosinophilic epidermolysis epididymitis, glomerulonephritis, The present invention relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the treatment of ulcerative colitis, Can be selected.

The present invention also provides a method of reducing or inhibiting the differentiation of undifferentiated T cells into Th17 cells in vitro, comprising treating undifferentiated T cells with a levamifeed compound or a pharmaceutically acceptable salt thereof. To provide.

In one embodiment of the present invention, the reduction or inhibition of differentiation of undifferentiated T cells into Th17 cells may be achieved through inhibition of production of IL-17 cytokines.

In addition, the present invention provides a method of activating regulatory T cells comprising the step of treating the LevamiFeid compound or a pharmaceutically acceptable salt thereof to Regulatory T cells (Treg) in vitro. to provide.

In one embodiment of the present invention, activated regulatory T cells may have increased expression of Foxp3.

Furthermore, the present invention provides a composition for immunosuppression comprising a levamifeed compound or a pharmaceutically acceptable salt thereof as an active ingredient.

The levamifeed compound according to the present invention has an excellent effect of inhibiting the differentiation of cytotoxic Th17 cells that produce and secrete inflammatory cytokines, and has the property of inhibiting the function of abnormally activated immune cells and controlling the inflammatory response. Since it has an excellent effect of enhancing the activity of immunoregulatory T cells (Treg), it is a pharmaceutical that can prevent or treat immune diseases such as autoimmune diseases, inflammatory diseases, and transplant rejection diseases caused by various cancer diseases or immune control disorders. It can be usefully used as a composition or an immunosuppressant. In addition, there is no toxicity and side effects to the drug can be used safely even when taking long-term, has a stable effect on the body.

1 shows the results of measuring the secretion amount of IL-17 cytokines produced after treatment of splenocytes isolated from mice under Th17 differentiation conditions and treatment with levamifeed for each concentration. The graph shown.
2 is a graph showing the results of measuring the secretion amount of IL-17 cytokines produced after treatment of human PBMC cells with Th17 differentiation conditions and treatment with levamifeed at each concentration in one embodiment of the present invention. .
Figure 3 is a graph showing the results of FACs stain performed to compare the expression of IL-17 and Tregs in mouse spleen cells 3 days after differentiation into Th17 cells according to an embodiment of the present invention.
Figure 4 is a graph showing the results of the FACs stain performed to compare the expression of IL-17 and Tregs in human PBMC cells 3 days after differentiation into Th17 cells according to an embodiment of the present invention.
5 is a Th17 differentiation environment when levami feed is treated at a concentration of 1000 μM using a mouse splenocyte model to investigate whether there is an action of activating Treg cells, which are immunoregulatory cells, according to an embodiment of the present invention. The expression level of IL-17, Foxp3, RORrt, STAT3, CCL20 and CCR6 in rough cells was analyzed by RT-PCR.
6a is a graph showing the therapeutic effect of arthritis through clinical index after oral administration of levamifeed 0.6mg / kg and 6mg / kg in collagen-induced arthritis animal model according to an embodiment of the present invention. to be.
Figure 6b is oral administration of levamifeed 0.6mg / kg, 6mg / kg, respectively, in the collagen-induced arthritis animal model according to an embodiment of the present invention, and then killed the animal model, joint and cartilage tissue The degree of destruction was observed through tissue staining.
Figure 7a shows the results confirmed by Western blotting analysis of phosphorylation changes of IL-6 dependent STAT3 by levamifeed in accordance with an embodiment of the present invention.
Figure 7b shows the results confirmed by flow cytometry changes in phosphorylation of TH17 cell differentiation induction STAT3 by levamifeed according to an embodiment of the present invention.

The present invention for the first time that the levamipid has the effect of preventing or treating cancer or immune diseases through the immunomodulatory action, and thus the present invention is a levamipid compound or a pharmaceutically acceptable salt thereof as an active ingredient It is characterized by providing a composition for the prevention or treatment of cancer or immune diseases, including.

The present inventors have focused on rebamipide compounds to develop new immunomodulators for the treatment of immune diseases, which have excellent effects in the treatment of gastric mucosal damage caused by acute exacerbation of gastric ulcers, acute gastritis or chronic gastritis. Eggplant is a medicine, widely used as a therapeutic agent for peptic ulcer, and its chemical name is 2- (4-chlorobenzoylamino) -3- [2 (1H) -quinolinone-4-yl] propionic acid. The drug protects gastric mucosa by promoting PGE2 biosynthesis to increase mucus, promotes cell proliferation, and inhibits bacterial adhesion and invasion to gastric mucosa cells, especially in patients infected with Helicobacter pylori. Suppresses stomach inflammation.

However, there is no mention in the prior art that levamipid can be used for the treatment of immune diseases or cancer diseases.

Therefore, in the present invention, the levamifeed compound may be used for preventing or treating cancer or immune disease, and in particular, levamifeed inhibits the differentiation of Th17 cells, which are pathogenic cells, and simultaneously regulates Treg cells (Regulatory T cells: Treg). It was first identified that the activity of) can induce a prophylactic or therapeutic effect of the immune disease or cancer disease through a mechanism for promoting it.

On the other hand, in general, the immune system controls specific immune responses to autoantigens in a normal state, and also suppresses immune responses to external antigens. For example, a pregnant woman's response to a fetus and a chronically infected microorganism are present. Immune response. These phenomena are known to be induced by clonal deletion, clone anergy and active control by immunoregulatory T cells (Treg) as a mechanism by which antigen specific immunotolerance can be induced. Investigating some patients who accidentally acquired immunotolerance against transplanted antigens or experimentally induced animal models showed that all three of these mechanisms are involved in immunological tolerance. Is attracting attention as an important cell involved in controlling almost all immune responses of living body such as autoimmune, tumoral immunity, infectious immune response as well as transplantation immune response.

In particular, immunoregulatory T cells, ie immunoregulatory T lymphocytes (Tregs), whose presence has recently been identified, can be largely divided into natural and adaptive Treg cells, and CD4 + CD25 + T cells, which are natural Tregs, are cells. Has been given immunosuppressive function when it is newly created in the thymus, and is present at a frequency of 5 to 10% of peripheral CD4 + T lymphocytes in normal individuals. Although the immunosuppressive mechanism of this cell has not yet been elucidated yet, it has recently been shown that the expression control gene of Foxp3 plays an important role in the differentiation and activity of this cell. In addition, peripheral natural T cells can be differentiated into cells that exhibit immunosuppressive effects upon stimulation of autologous or external antigens under certain circumstances, which are called adaptive or inducible Tregs and secrete IL-10. Tr1, Th3 and CD8 Ts that secrete TGF-ß, and the like.

In addition, as described above in the prior art, T cells are also differentiated into Th17 cells through differentiation in addition to Treg cells. Th17 cells are formed in the presence of TGF-ß in common with Treg cells, but in the case of Treg cells, IL- On the other hand, Th17 cells are characterized by differentiating in the presence of IL-6 with TGF-ß and secreting IL-17.

In addition, Th17 cells have cytotoxic properties that maximize signal of inflammatory response and accelerate disease progression. Thus, inhibition of differentiation or activity into Th17 cells is one of the ways to treat cancer or immune diseases.

Accordingly, the present inventors have determined whether the levamifeed compound of the present invention can treat immune or inflammation-related diseases and cancer diseases by regulating T cells, specifically, differentiating T cells into Treg cells and Th17 cells. According to one embodiment of the present invention, the splenic cells isolated from the mouse and human PBMC cells are treated with stimulation conditions that can differentiate the undifferentiated T cells present in the cells into Th17 cells and at the same time rebam Feed compounds were treated and cultured at each concentration, and then the amount of IL-17 cytokine expressed in the cells was measured. In general, the inflammatory cytokine IL-17 is produced from Th17 cells and can measure the differentiation of undifferentiated T cells into Th17 cells through the amount of IL-17 produced.

As a result, in the control group not treated with levami feed, Th17 cells were differentiated by Th17 cell differentiation conditions, and the production of IL-17 cytokines was significantly increased compared with the group without any treatment, whereas levami In the feed-treated group, the production of IL-17 cytokines was significantly reduced compared to the non-levamifeed-treated group. In addition, the amount of IL-17 cytokine produced was found to decrease in proportion to the concentration of treated levamifeed (see FIGS. 1 and 2).

Accordingly, the present inventors have found that the Levamifeed compound reduces or inhibits the differentiation of the undifferentiated T cells into Th17 cells, which are pathogenic cells that produce the inflammatory cytokine IL-17.

Therefore, the levamifeed compound of the present invention is characterized in that it can prevent or treat cancer or immune diseases through the action of reducing or inhibiting the differentiation of undifferentiated T cells into Th17 cells, which are pathological cells. That is, the present invention can prevent or treat cancer or immune disease by suppressing an immune response or an inflammatory response.

Furthermore, the present inventors measured the expression level of Foxp3 gene according to the treatment of levamipid in order to investigate how levamipid affects the activity or proliferation of immunoregulatory T cells (Treg).

Foxp3 is mainly present in regulatory T cells derived from the thymus and is a transcriptional factor present in cells with CD4 + CD25 + labeling antigen, and its function is to express Foxp3. Recognition of IL-2 against T cells that are potentially responsive to antigens and that may potentially cause autoimmune among CD4 + CD25- T cells that do not express Foxp3 differentiated from the thymus at the time of antigen recognition against T cells It has a role as a suppressor T cell that suppresses production and cell division. In addition, Foxp3 stimulates not only IL-2 but also IL-4, which is influenced by NFAT, which is a transcription factor, in CD25- T cells through regulatory T cells expressing Foxp3 and cell-cell contact with them, Lt; RTI ID = 0.0 > IFN-gamma. ≪ / RTI > Therefore, in the case of T cells expressing Foxp3, which has the above-mentioned action, it is applied to the treatment of immune diseases through the action of suppressing or modulating the immune response, and it is also applied to the treatment of CD4 T cells expressing Foxp3 To increase the number of autoantigen-specific T cells (self-antigen specific T cell clone) by treatment with high concentration of IL-2 cytokine and combination treatment with anti-CD3 and anti-CD28 antibody, There have been attempts to. Thus, confirmation of expression of Foxp3 is an indicator of the activity or amplification of Treg cells.

According to another embodiment of the present invention, the splenocytes isolated from the mouse are treated with stimulation conditions capable of differentiating into Th17 cells and simultaneously treated with levamipid and then cultured, and then the degree of amplification or activity of differentiated Treg cells. As a result of measuring and analyzing the amount of Foxp3 expressed, the amount of Foxp3 mRNA expressed in Treg cells was significantly increased in the group treated with levamifeed compared to the group without levamifeed. The increase was shown to be proportional to the concentration of treated levamifeed. In addition, the transcription factors RORrt and STAT3, which are involved in the differentiation of Th17 cells, and CCL20 and CCR6, which are migration markers of Th17 cells, were also reduced by treatment with levamifeed (see FIG. 5).

Therefore, the present inventors have found that the levamifeed compound can be used to prevent or treat cancer or immune diseases by activating or amplifying regulatory T cells (Treg).

Furthermore, in one embodiment of the present invention, it was investigated whether levamipid can cure an immune disease in an animal model, and arthritis in a group administered with levamipid and a group not administered to an arthritis animal model, respectively. As a result of analyzing the degree, the arthritis symptom was significantly improved in the levamifeed-administered group compared to the control group (see FIGS. 6A and 6B).

On the other hand, STAT (Signal transducers and activators of transcription) is a signal transduction and transcription regulator protein, is activated by extracellular stimulation of cytokines, hormones, growth factors, etc., phosphorylation of tyrosine residues, dimers by the interaction of the SH2 domain A dimer is formed that enters the nucleus and binds to a specific promoter. This STAT protein signaling system can be inhibited by dephosphorylation and protein degradation.

Recently, STAT3, STAT3, and STAT5 have been found to be active forms in various carcinomas. STAT3 is not only a blood cancer such as leukemia, but also breast cancer, head and neck cancer, melanoma, ovarian cancer, lung cancer, pancreatic cancer, prostate cancer It is activated in a variety of solid tumors and has become an important cancer target (Hua Yu and Richard Jove, Nature Review Cancer ., 2004, 8, 945).

In addition, STAT3 activity has been known to inhibit apoptosis, induce angiogenesis, and induce immune evasion (Wang T. et al., Nature Medicine ., 2004, 10, 48). Therefore, inhibition of STAT3 activity has an effect of controlling the tumor by a complex anticancer mechanism. STAT3 protein is involved not only in tumor but also various cellular functions, so that its inhibitor can be developed as an immunosuppressant. Therefore, if the activity of STAT3 can be inhibited, diseases such as cancer and immune diseases can be treated.

In order to investigate whether the levamifeed compound of the present invention can increase the phosphorylation during differentiation of TH17 cells that secrete cytokines such as IL-6 and inflammatory cytokines, it can inhibit the activity of activated STAT3. Western blot and flow cytometry analysis of the tissues of the mouse model treated with levamifeed showed a significant decrease in phosphorylation of STAT3 in the group treated with levamifeed compared to the group without treatment. 7a and 7b, the results indicate that the inventors can treat the cancer or immune disease through the mechanism of inhibiting the phosphorylation of STAT3.

Therefore, the levamifeed compound of the present invention inhibits or reduces the differentiation of undifferentiated T cells into Th17 cells secreting inflammatory cytokines and / or activates or amplifies regulatory T cells that can normally regulate excessive immune responses. It is characterized by being able to prevent or treat diseases related to immunity or inflammation and cancer diseases through promoting or increasing action.

In the present invention, the term "active" means that all the mechanisms of a regulatory T cell (Treg), that is, a Treg cell including both natural and adaptive Treg cells, are promoted or promoted in vivo. It means that the immunomodulatory action such as an immunosuppressive reaction is promoted or promoted so that the immune response in the body is in a steady state.

In addition, the term "expansion" refers to the differentiation and proliferation of undifferentiated T cells into regulatory T cells, and 'differentiation' refers to a phenomenon in which structures or functions are specialized while the cells divide and grow, In other words, the shape or function of a cell or tissue of an organism is changed in order to perform a given task, and 'proliferation' refers to the division of cells and the expansion of homogeneous ones. It means to increase.

Therefore, the present invention can provide a composition for the prevention or treatment of cancer or immune diseases comprising levamipid or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the levamipid according to the present invention may be a compound represented by the following formula (1).

≪ Formula 1 >

In addition, the rebamipide compound according to the present invention can be used in the form of a salt, preferably a pharmaceutically acceptable salt. The salt is preferably an acid addition salt formed by a pharmaceutically acceptable free acid, and the free acid may be an organic acid or an inorganic acid. The organic acids include, but are not limited to, citric, acetic, lactic, tartaric, maleic, fumaric, formic, propionic, oxalic, trifluroacetic, benzoic, gluconic, methosulfonic, glycolic, succinic, Glutamic acid and aspartic acid. The inorganic acid includes, but is not limited to, hydrochloric acid, bromic acid, sulfuric acid, and phosphoric acid.

The levamifeed compound according to the present invention may be used that is isolated from nature or prepared by chemical synthesis known in the art.

The levamitide compound according to the present invention can reduce or inhibit Th17 cells, which are pathological cells, while preventing or treating cancer or immune diseases through the action of promoting or enhancing the activity or amplification of regulatory T cells.

In the present invention, "cancer" is defined as a disease characterized by the fast and uncontrolled growth of abnormal cells. Cancer cells can spread locally or through the bloodstream and lymphatic system to other parts of the body. In addition, the type of cancer in the present invention is not limited to this, colon cancer, rectal cancer, anal muscle cancer, bone cancer, gastric cancer, cerebrospinal tumor, head and neck cancer, thymic tumor, mesothelioma, esophageal cancer, biliary tract cancer, bladder cancer, testicular cancer, small intestine cancer , Germ cell tumor, endometrial cancer, fallopian tube carcinoma, vaginal carcinoma, vulvar carcinoma, multiple myeloma, sarcoma, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, bladder cancer, urethral cancer, pituitary adenoma, renal pelvic carcinoma, spinal cord tumor, multiple myeloma , Glioma, CNS central nervous system tumor, hematopoietic tumor, fibrosarcoma, neuroblastoma, astrocytoma, breast cancer, cervical cancer, ovarian cancer, prostate cancer, pancreatic cancer, kidney cancer, liver cancer, brain cancer, lung cancer, lymphoma, leukemia , Malignant melanoma, skin cancer, and the like.

As used herein, the term "immune disease" refers to a disease in which components of the mammalian immune system cause, mediate or otherwise contribute to the pathology of a mammal. In addition, the stimulation or discontinuation of the immune response may include all diseases that have a compensatory effect on the progress of the disease, and the present invention may include diseases caused by an irritable immune response. Examples of such immune disorders include, but are not limited to, autoimmune diseases; Inflammatory disease; And transplant rejection of cells, tissues, or organs.

In addition, one of the most important properties of all normal individuals is that they do not react negatively to the antigenic material that constitutes the self, while non-self antigens are recognized and reacted to remove I have the ability. Such a non-response of a living body to a self-antigen is called immunologic unresponsiveness or tolerance.

However, when there is a problem in inducing or maintaining such self-tolerance, an immune response occurs to autoantigens, and this causes an attack on one's own tissue. The disease caused by this process is called an "autoimmune disease". It is called.

In addition, an "inflammatory disease" refers to a tumor necrosis factor-a (TNF-a) or IL-1 (excessively stimulated by the immune system such as macrophages due to excessive stimulation of the human immune system due to harmful stimuli such as inflammation-inducing factors or irradiation). interleukin-1), IL-6, prostaglandin (prostagladin), luecotriene or nitric oxide (nitric oxide, NO) refers to a disease caused by proinflammatory substances (inflammatory cytokines).

On the other hand, in order to successfully transplant the organ, the recipient's immune rejection response to the transplanted cells and organs must be overcome. The main mediator of transplantation immune rejection is T cell, which is expressed in the graft. Major histocompatibility complex (MHC) is recognized by T cell receptor (T cell receptor) A reaction occurs. Immunosuppressive agents have been used to reduce these transplant immune rejection responses. A common goal of these immunosuppressive agents is to inhibit T cell-mediated immune responses to grafts. Recently, regulatory T cells have been used to suppress immune responses, Methods to treat refractory diseases have been attempted.

In addition, the immunological diseases of the present invention include, but are not limited to, Behcet's disease, multiple myositis / dermatomyositis, autoimmune hemodilution, autoimmune myocarditis, atopic dermatitis, asthma, primary cirrhosis, dermatomyositis, Autoimmune hepatitis, autoimmune mumps, Crohn's disease, insulin-dependent diabetes mellitus, eosinophilic epidermolysis, epididymitis, glomerulonephritis, autoimmune thrombocytopenia, autoimmune thrombocytopenia, autoimmune meningitis, Sjogren's syndrome, systemic lupus erythematosus, Rheumatic fever, rheumatoid arthritis, sarcoidosis, scleroderma, spondyloarthropathies, thyroiditis, vasculitis, vitiligo, mucinous anemia, hemolytic anemia, multiple sclerosis, myasthenia gravis, pemphigus vulgaris, , Ulcerative colitis, and the like.

Therefore, the composition according to the present invention can be used as a pharmaceutical composition that can prevent or treat cancer or immune diseases.

The term “treatment”, unless stated otherwise, means to reverse, alleviate, inhibit the progression of, or prevent the disease or condition to which the term applies, or one or more symptoms of the disease or condition. As used herein, the term "treatment" refers to the act of treating when "treating" is defined as above. Thus, in mammals, "treatment" or "therapy" of cancer or immune disease may include one or more of the following:

(1) inhibits the growth of cancer or immune disease, ie inhibits its development,

(2) preventing the spread of cancer or immune disease, ie preventing metastasis,

(3) reduce cancer or immune disease.

(4) prevent the recurrence of cancer or immune disease, and

(5) Palliating the symptoms of cancer or immune disease

The composition for preventing or treating cancer or immune disease according to the present invention may include a pharmaceutically effective amount of a levamifeed compound or a salt thereof alone or may include one or more pharmaceutically acceptable carriers, excipients or diluents. . The pharmaceutically effective amount herein refers to an amount sufficient to prevent, ameliorate and treat the symptoms of cancer or immune disease.

The pharmaceutically effective amount of the levamifeed compound or salt thereof according to the present invention is 0.5 to 100 mg / day / kg body weight, preferably 0.5 to 5 mg / day / kg body weight. However, the pharmacologically effective amount may be appropriately changed depending on the severity of the immune disease symptoms, the age, body weight, health condition, sex, administration route and treatment period of the patient.

Also, the pharmaceutically acceptable hereinabove refers to a composition that is physiologically acceptable and does not normally cause an allergic reaction such as gastrointestinal disorder, dizziness, or the like when administered to a human. Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. Further, it may further include a filler, an anticoagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent and an antiseptic agent.

In addition, the compositions of the present invention may be formulated using methods known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal. The formulations may be in the form of powders, granules, tablets, emulsions, syrups, aerosols, soft or hard gelatine capsules, sterile injectable solutions, sterile powders.

In addition, the composition for preventing or treating cancer or immune disease according to the present invention may be administered through various routes including oral, transdermal, subcutaneous, intravenous or intramuscular, and the dosage of the active ingredient is determined by the route of administration, the age of the patient, It may be appropriately selected according to various factors such as sex, weight and severity of the patient, and the composition for preventing or treating cancer or immune disease according to the present invention has the effect of preventing, improving or treating the symptoms of cancer or immune disease. It can be administered in combination with a known compound.

Accordingly, the present invention can provide a medicament for the prevention or treatment of cancer or immune diseases, including a composition containing a levamifeed compound or a salt thereof as an active ingredient, and the present invention further provides a levamifeed compound or a salt thereof. It is possible to provide an immunosuppressive composition comprising as a component.

In addition, the present invention can provide a method for reducing or inhibiting the differentiation of undifferentiated T cells into Th17 cells in vitro, comprising treating the undifferentiated T cells with a levamifeed compound or salt thereof. The reduction or inhibition of the differentiation of the undifferentiated T cells into Th17 cells may be achieved through the inhibition of production of IL-17 cytokines.

In addition, the present invention can provide a method for activating regulatory T cells comprising the step of treating the levamifeed compound or salt thereof to Regulatory T cells (Treg) in vitro, The regulatory T cells activated according to the present invention may increase the expression of Foxp3.

The method of reducing or inhibiting the differentiation of undifferentiated T cells into Th17 cells in vitro and the method of activating regulatory T cells in the above-described method according to the present invention are as described above. The levamifeed compound may be directly treated to the medium for culturing cells, or the composition containing the levamifeed compound according to the present invention as an active ingredient may be treated to the culture medium. In addition, at this time, the concentration of the levamifeed compound that can be treated in the culture medium may be treated so that the final concentration is 50μM ~ 1500μM, in one embodiment of the present invention the concentration of levamifeed compound is the final 100μM, 250μM, 500 μM and 1000 μM were added to the culture medium of the cells.

Furthermore, the composition for the prevention or treatment of cancer or immune disease according to the present invention is excellent in the activity or amplification effect of immune regulatory T cells (Treg), and excellent in inhibiting the differentiation of Th17 cells, which are pathogenic cells producing inflammatory cytokines. In addition, there is no toxicity and side effects for the drug, so it can be used safely even when taken for a long time, and has a stable characteristic for the body.

Therefore, the present invention can provide a food composition which can improve or prevent the symptoms of cancer or immune diseases containing the levami feed compound or salt thereof as an active ingredient, the food composition according to the invention is cancer or immune Food, which is effective for improving or preventing disease symptoms, for example, it can be easily utilized as a main ingredient, side ingredients, food additives, functional food or beverage of food.

As used herein, the term "food" means a natural product or a processed product containing one or more nutrients, preferably a state capable of being directly eaten through a certain degree of processing, , Food, food additives, functional foods and beverages.

Foods to which the food composition according to the present invention may be added include, for example, various foods, beverages, gums, teas, vitamin complexes, functional foods, and the like. In addition, in the present invention, the food may include special nutritive foods (e.g., crude oil, spirits, baby food, etc.), meat products, fish meat products, tofu, mackerel, noodles (Such as soy sauce, soybean paste, kochujang, mixed potatoes), sauces, confectionery (eg, snacks), candies, chocolate, gums, ice cream, milk products (eg, fermented milk, cheese, But are not limited to, pickled foods (various kinds of kimchi, pickles, etc.), beverages (e.g., fruit drinks, vegetable beverages, beverages, fermented beverages and the like) and natural seasonings (e.g. The food, beverage or food additive may be prepared by a conventional production method.

The above-mentioned "functional food" refers to a food group which is imparted with added value to function and express the function of the food by physical, biochemical or biotechnological techniques, or to control the bio-defense rhythm of the food composition, Refers to a food prepared by processing a body so as to sufficiently express the body's control function on the body, such as recovery, and the like. Specifically, it may be a health functional food. The functional food may include a food-acceptable food-aid additive, and may further comprise suitable carriers, excipients and diluents conventionally used in the production of functional foods.

In addition, in the present invention, the term "beverage" refers to a generic term for drinking to quench thirst or to enjoy a taste and includes a functional beverage. The beverage contains, as the essential ingredients, the composition for improving or preventing the symptoms of cancer or immune disease as an essential ingredient, and there are no special limitations to the other ingredients, and various flavors or natural carbohydrates, such as ordinary drinks, may be added. It may contain as.

Furthermore, in addition to the foregoing, foods containing food compositions for improving or preventing symptoms of cancer or immune diseases of the present invention may be flavored with various nutrients, vitamins, minerals (electrolytes), synthetic flavors, and natural flavoring agents. , Colorants and fillers (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. It may contain, the components can be used independently or in combination.

In the food containing the composition for food of the present invention, the amount of the composition according to the present invention may comprise from 0.001% to 90% by weight of the total food weight, preferably from 0.1% to 40% by weight In the case of a beverage, it may be included in a ratio of 0.001g to 2g, preferably 0.01g to 0.1g based on 100ml, in the case of long-term intake for health and hygiene purposes or health control purposes It may be less than the above range, and the active ingredient is not limited to the above range because it may be used in an amount above the above range because there is no problem in terms of safety.

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited to these embodiments.

< Example  1>

Th17  In a cell Levamifeed  by IL -17 Production inhibitory activity assay

The present inventors investigated whether the levamifeed compound is active in inhibiting the production of the inflammatory cytokine IL-17 in Th17 cells, which are pathogenic cells.

<1-1> Th17  Induction of cell differentiation and Levamifeed  process

① mouse splenocytes: Spleen cells were obtained from the mouse, that is, the spleens extracted from the mice were finely chopped of the spleen tissue using a glass slide (teasing slide), and then red blood cells in the spleen were removed with erythrocyte hemolysis solution. Splenocytes were then obtained by adding PBS buffer solution and washing by centrifugation.

Then, 1 × 10 ⁶ single cells isolated from the spleen were dispensed into a 24-well plate coated with 1 μg / ml of anti-CD3 antibody, and anti-CD28 antibody under conditions capable of stimulating Th17 cells. Treatment of 1 μg / ml, TGF-ß 20ng / ml, IL-6 20ng / ml, anti-IL-4 10ng / ml, anti-IFNr 10ng / ml together and incubated cells for 3 days to differentiate Th17 cells Induced. At this time, the cells were incubated by treating the levamifeed compound at concentrations of 250 μM, 500 μM, and 1000 μM, respectively.

② Human cell lines: Peripheral blood mononuclear cells (PBMCs) isolated from human peripheral blood (PBMC) 1 × 10 ⁶ were dispensed into 24 well plates coated with anti-CD3 antibody coated with 1 μg / ml, and Th17 cells Anti-CD28 antibody 1μg / ml, IL-6 20ng / ml, IL-1ß 20ng / ml, IL-23 20ng / ml, anti-IL-4 10ng / ml, anti-IFNr 10ng / After simultaneous treatment with ml, cells were cultured for 3 days to induce Th17 cell differentiation. At this time, the cells were incubated by treating the level of levamifeed compounds at concentrations of 100 μM and 500 μM, respectively.

<1-2> IL -17 cytokine measurement ( ELISA )

To measure the amount of IL-17 cytokine produced, the cell-cultured supernatant was collected and examined for IL-17 expression using IL-17 and sandwich ELISA. The monoclonal anti-IL-17 was reacted overnight at 4 ° C. at 2 μg / ml in a 96-well plate, and non-specific binding was blocked with a blocking solution (1% BSA / PBST) after the reaction. IL-17 recombinant was serially diluted by 1/2 and used as a standard. Cell culture supernatant was added and reacted at room temperature for 2 hours. Thereafter, the biotinylated anti-IL-17 was reacted at room temperature for 2 hours and washed four times. Then, the ExtraAvidin-Alkaline Phosphatase conjugate was diluted and added and reacted at room temperature for 2 hours. Thereafter, PNPP / DEA solution was added, followed by color development, and absorbance at 405 nm was measured.

As a result, in the mouse splenocyte model, as shown in FIG. 1, IL-17, an inflammatory cytokine produced from Th17 cells, which are pathogenic cells, was found to be inhibited by the levamifeed compound of the present invention. It has been shown that the production of IL-17 is reduced depending on the level of levamifeed.

On the other hand, in the case of the human cell line model, as shown in FIG. 2, IL-17, an inflammatory cytokine produced from pathogenic cells, Th17 cells, was shown to be inhibited in production by the levamipid compound of the present invention. It was shown that the production of IL-17 is reduced depending on the mifeed concentration.

< Example  2>

FACs stain  analysis

Three days after differentiation into Th17 cells, FACs staining was performed to compare the expression of IL-17 and Tregs in the cells.

<2-1> mouse Splenocyte

After collecting and washing the cells with FACs buffer, the reaction was blocked for 5 minutes at 4 ℃ to inhibit non-specific binding and washed with FACs buffer. Anti CD4 percp cy5.5 and anti CD25 APC were added and reacted at 4 ° C. for 30 minutes and washed with FACs buffer. 100ul each of Cytoperm / cytofix buffer was added and reacted at 4 ° C. for 30 minutes and washed with perm wash buffer. To inhibit nonspecific binding, the blocking was reacted at 4 ° C. for 15 minutes and then washed with perm wash buffer. Anti IL-17 FITC and anti Foxp3 PE were added and reacted at 4 ° C. for 30 minutes and washed with perm wash buffer. The stained cells were washed with FACs buffer and analyzed using a flow cytometer (FACs, fluorescent-activated cell sorter).

As a result, as shown in Figure 3, when treated with 250μM levamifeed, Foxp3 expression was 11.6, IL-17 expression was 28.9, and when treated with 1000μM levamifeed, Foxp3 expression was 20.2 , IL-17 expression was 15.7, while untreated control group Foxp3 expression was 9.81, IL-17 expression was 25.5. These results confirm that Foxp3 expression gradually increases and IL-17 expression decreases depending on the treatment concentration according to the treatment of levamifeed.

<2-2> human PBMC  cell

After collecting the cells and washing with FACs buffer, the reaction was blocked for 15 minutes at 4 ℃ to inhibit the non-specific binding and washed with FACs buffer. Anti CD4 percp cy5.5 and anti CD25 APC were added and reacted at 4 ° C. for 30 minutes and washed with FACs buffer. 100ul each of cytotom / cytofix buffer was added and reacted at 4 ° C. for 30 minutes, and then washed with perm wash buffer. Then, anti Foxp3 PE was added and reacted at 4 ° C. for 30 minutes, followed by washing with perm wash buffer. The stained cells were washed with FACs buffer and analyzed using a flow cytometer (FACs, fluorescent-activated cell sorter).

As a result, as shown in Figure 4, Foxp3 expression was 3.22 when treated with 100μM levamipid, 5.64 when treated with 500μM levamifeed, whereas untreated control was 2.59. These results confirmed that the expression of Foxp3 gradually increased depending on the treatment concentration according to the treatment of levamipid.

< Example  3>

Levamifeed Th17  Cells and Treg  Analysis of effects on cells RT - PCR )

Controlled T lymphocytes, or Treg (regulatory T lymphocytes) cells, are known to play a key role in the maintenance of immune tolerance (Nat. Rev. Immunol. 2009, 7: 305) and are produced primarily in the thymus and are transcription factors Expression of Foxp3 is required and Foxp3 is known to play a role in inhibiting the production of inflammatory cytokines. In addition, RORrt and STAT3 are known as transcription factors involved in differentiating Th17 cells, and CCL20 and CCR6 are known as Th17 migration markers.

In order to investigate whether the levamifeed compound according to the present invention has an action of activating the activity of Treg cells, which are immunoregulatory cells, the cells undergoing the Th17 differentiation environment of Example 1 using a mouse splenocyte model RT-PCR was performed to analyze the expression levels of IL-17, Foxp3, RORrt, STAT3, CCL20 and CCR6.

First, levamipid was treated to 1000 μM of cells for Th17 differentiation conditions, and then RNAs were extracted from the cells according to methods known in the art, respectively, to obtain cDNA products. The polymerase chain reaction was performed using the primers shown in Table 1 below to amplify IL-17, Foxp3, RORrt, STAT3, CCL20, CCR6 and ß-actin as a control using the obtained cDNA product. The reaction compound for RT-PCR was brought to a total of 25 μl, and the composition of the solution was used for each primer pair having 2.5 μl of 10 × reaction buffer 0.5 mM dNTP and the sequence described in Table 1 below. For the amplification, the reaction was repeated 25 times for a denaturation step of 94 ° C. 30 seconds, annealing step 60 ° C. 30 seconds, and extension step 72 ° C. 30 seconds using a dual-bay thermal cycler system instrument. Is shown in Table 1 below.

Primer sequence primer Primer sequence SEQ ID NO: IL-17 sense  5'-CCTCAAAGCTCAGCGTGTCC-3 ' One IL-17 antisense  5'- GAGCTCACTTTTGCGCCAAG-3 ' 2 Foxp3 sense  5'-CCCAGGAAAGACAGCAACCTT-3 ' 3 Foxp3 antisense  5'-CAAACAGGCCGCCGTCTGGAGCC-3 ' 4 RORrt sense  5'-TGTCCTGGGCTACCCTACTG-3 ' 5 RORrt antisense  5'- GTGCAGGAGTAGGCCACATT-3 ' 6 STAT3 sense  5'-GACCCGCCAACAAATTAAGA-3 ' 7 STAT3 antisense  5'-TCGTGGTAAACTGGACACCA-3 ' 8 CCL20 sense  5'-CAGCTGTTGCCTCTCGTACA-3 ' 9 CCL20 antisense  5'-CACCCAGTTCTGCTTTGGAT-3 ' 10 CCR6 sense  5'-CCATGACTGACGTCTACCTGTTGAACA-3 ' 11 CCR6 Antisense  5'-GAACAGCTCCAGTCCCATACCCAGCAG-3 ' 12 ß-actin sense  5'-GAAATCGTGCGTGACATCAAAG-3 ' 13 ß-actin antisense  5'-TGTAGTTTCATGGATGCCACAG-3 ' 14

As a result, as shown in Figure 5, when the level of levamifeed treated with 1000μM concentration of IL-17 in the cells was shown to be significantly reduced compared to the control, Foxp3 expression was increased compared to the control appear. In addition, transcription factors RORrt and STAT3, which are involved in the differentiation of Th17 cells, and CCL20 and CCR6, which are migration markers of Th17 cells, were also decreased by treatment with levamifeed.

< Example  4>

In an arthritis animal model Levamifeed  Analysis of treatment effect according to administration

<4-1> Preparation of Animal Model

The test animal was a 7-week-old male DBA / 1J mouse, and in order to prepare an arthritis animal model, type 2 collagen (Cll) was dissolved in 0.1 N acetic acid solution to 4 mg / ml, followed by dialysis buffer, Dialysis with 50mM Tris, 0.2N Nacl) and mixed in the same amount with complete Freund's adjuvant (CFA, Chondrex) containing M. tuberculosis and subcutaneously injected at the base of the tail of the mouse to 100μL per animal Ie 100 μl / 100 μl) (first injection). Two weeks later, the same CII was mixed with the same amount of incomplete Freud's adjuvant (IFA, Chondrex) and then 100 μl (ie 100 μl / 100 μl) was injected into one hind limb (foot pad) (second injection).

After the second immunization through the second injection, 10 mg of 0.6 mg / kg and 6 mg / kg of levamipids were injected into the oral cavity three times a week, respectively, wherein the levamifeed was dissolved in 0.5% CMC solution and used in each group. Were conducted in 5 rats and arthritis was evaluated up to 10 weeks. In addition, each animal was killed at a time when the arthritis index was significantly different for an in vitro test, and the effect of arthritis disease in the blood and joint tissues and the therapeutic effect by levamifeed were as follows. Investigate.

<4-2> Arthritis in Animal Models Levamifeed  by Rheumatism  Arthritis Treatment Activity Assessment

<4-2-1> Joint Index Evaluation

Three weeks after the first inoculation, four observers, who did not know the contents of the experiment, evaluated the severity of the joint inflammation three times a week and observed up to 10 weeks. At this time, arthritis evaluation was averaged by dividing the scores of the three legs except the legs to which CII / CFA was administered at the second dose per dividing according to the following scale, divided by three, and then obtained by three observers in each animal model. The average value divided by summing was used. The scores and criteria according to arthritis evaluation are as follows.

-Evaluation standard-

0 points: There is no edema or swelling.

1 point: mild edema and redness limited to the foot or ankle joint

2 points: Mild swelling and redness from the ankle joint to the metatarsal

3 points: moderate swelling and redness from the ankle joint to the ankle bone

4 points: swelling and redness from ankle to leg

The best arthritis index per head is 4, so the best disease index per mouse is 16.

Observations showed that the collagen-induced arthritis mouse model continued to increase arthritis scores over four weeks, resulting in severe arthritis symptoms, whereas the arthritis index decreased in animal models injected with levamifeed, especially Leva Mice injected with mipid at an amount of 6 mg / kg showed little increase in arthritis index (see FIG. 6A).

<4-2-2> Histological Examination

The collagen-induced arthritis (CIA) animal model was orally administered with levamipid at doses of 0.6 mg / kg and 6 mg / kg, respectively. After 10 weeks, the animals were euthanized, and the hind paws of the mice were fixed with 10% formalin, decalcified from bone, and paraffin-embedded. Joint sections (7 um) were prepared and stained with hematoxylin and eosin. In addition, histological examination was performed by toluidine blue and safranin O staining to confirm the degree of cartilage destruction.

As a result of histological examination, the joints of CIA animals were infiltrated with many immune cells, and the formation of pannus, cartilage destruction and bone erosion were observed. On the other hand, animals given oral administration of 0.6 mg / kg or 6 mg / kg of levamifeed were observed to have a similar degree of destruction of joints and cartilage and remained similar to normal mice (see FIG. 6B).

<4-2-3> Western Blot  Analysis

Splenocytes were obtained from the mouse, and CD4 + T Cell 1 × 10 ⁷ isolated from the spleen was pretreated at a concentration of 1000 μM of levamifeed for 1 hour, and then stimulated for 1 hour at a concentration of 20ng / ml of IL-6. 20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS, 0.02% sodium azide, 1 mM phenylmethysulfonyl fluoride and 5 μg / mL leupeptin After homogenization and centrifugation at 13,000 rpm for 15 minutes, proteins were measured by Bradford method (Bio-Rad, Hercules, CA, USA). The quantified protein was mixed with the same extraction solution to which 4% SDS and 10% mercaptoethanol were added at a predetermined ratio, and then boiled at 100 ° C. for 10 minutes. 50 μg of protein was loaded on 10% SDS-PAGE gel, followed by electrophoresis, and then blotting onto nitrocellulose membrane using the electrical properties of the protein. 5% skim milk powder (in 0.05% Tween-20, PBS) was used to eliminate non-specific immune responses, phospho and total STAT3 or ß-actin primary and goat anti-rabbit IgG or goat anti-mouse IgG After treatment with the antibody, the degree of protein expression was observed using enhanced chemiluminescense (Amersham, Burlingame, CA, USA).

As a result, it was confirmed that the phosphorylated form of STAT3 increased by IL-6 was reduced in the degree of phosphorylation by treatment with levamifeed (see FIG. 7A).

<4-2-4> Flow cell  analysis

In order to stimulate 1 × 10 ⁶ of CD4 + T cells isolated from the spleen under Th17 cell differentiation conditions, an anti-CD3 antibody may be dispensed into a 24-well plate coated with 1 μg / ml and stimulate Th17 cells. Treatment with 1 μg / ml of anti-CD28 antibody, TGF-ß2ng / ml, IL-6 20ng / ml, anti-IL-4 10ng / ml, and anti-IFNr 10ng / ml was performed simultaneously. Stimulate At this time, the cells were incubated with pre-treatment of the levamifeed compound at a concentration of 1000 μM for one hour. The collected cells were loaded with PerCPμ-anti-mouse CD4 antibody and PE-anti-mouse phospho STAT3 antibody and reacted in the dark field for 30 minutes at 4 ° C, and washed with FACs buffer (0.002% sodium azide, 0.2% BSA / PBS). The cells were washed twice with Permwash buffer and resuspended in FACs buffer for analysis by flow cytometry.

Flow cytometry showed changes in the phosphorylation of Th17 cell differentiation-induced STAT3 by levamifeed. The phosphorylation of STAT3 increased during Th17 differentiation.

Finally, through the above results, the present inventors confirmed that the levamifeed compound according to the present invention inhibits cell differentiation into the pathogenic cell Th17, whereas the expression of Foxp3 has an activity of increasing levamifeed. Has been shown to be able to prevent or treat immune diseases or cancer diseases by activating Treg cells, which are immunoregulatory cells. It was found that the disease could be cured.

So far I looked at the center of the preferred embodiment for the present invention. Those skilled in the art will appreciate that the present invention can be implemented in a modified form without departing from the essential features of the present invention. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

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Claims (11)

delete delete delete delete delete delete A method of reducing or inhibiting differentiation of undifferentiated T cells into Th17 cells in vitro, comprising treating undifferentiated T cells with a rebamipide compound or a pharmaceutically acceptable salt thereof. The method of claim 7, wherein
Reduction or inhibition of differentiation of undifferentiated T cells into Th17 cells is achieved through the inhibition of production of IL-17 cytokines, which reduces or inhibits the differentiation of undifferentiated T cells into Th17 cells in vitro. Way. A method of activating regulatory T cells comprising treating a rebamipide compound or a pharmaceutically acceptable salt thereof in a regulatory T cell (Treg) in vitro. 10. The method of claim 9,
Activated regulatory T cells are a method of activating regulatory T cells, characterized in that the expression of Foxp3 is increased. delete

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