KR20170088671A - Composition for preventing or treating autoimmune disease comprising TWEAK inhibitor - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating autoimmune diseases, comprising a TWEAK inhibitor as an active ingredient. The TWEAK inhibitor according to the present invention can effectively treat autoimmune diseases such as arthritis and lupus by inhibiting signal transduction of TWEAK through targeting B cells.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for preventing or treating an autoimmune disease that contains a TWEAK inhibitor,

The present invention relates to a composition for preventing or treating an autoimmune disease containing a TWEAK inhibitor.

The immune system plays a role in protecting the body from harmful foreign substances. These types of antigens include bacteria, viruses, toxins, cancer cells and blood and tissues from other people or animals. The immune system produces antibodies to destroy these harmful substances. When an autoimmune disorder occurs, the immune system fails to differentiate between organs and harmful antigens of its healthy body, destroying normal tissues. The disease that causes is autoimmune disease.

If an autoimmune abnormality occurs, a reaction occurs to the normal tissues of the body. The reason why the immune system can not distinguish between the organ of the body and the antigen is that a microorganism such as bacteria or a drug has a gene that is susceptible to autoimmune diseases The hypothesis is that it causes illness in people born.

Treatment of autoimmune reactions is aimed at regulating the autoimmune response and restoring the damaged immune function of the body, which varies according to the type of autoimmune disease. For example, blood transfusions should be performed if there is a problem with the blood, and exercise or other functional treatment should the bones, joints, or muscles develop. Drugs are also prescribed to control the immune system's response. Immunosuppressive medicines are known to be used as immunosuppressant drugs, including corticosteroids, prednisone, cyclosteramide, cyclophosphamide, azathioprine, , And tacrolimus.

However, since these therapeutic agents cause side effects in the body and their therapeutic effects are not excellent, it is necessary to develop a new therapeutic agent for autoimmune disease which is more safe and effective.

TNF-related weak inducer of apoptosis (TWEAK) is an apoptosis derivative. It is a kind of ligand belonging to TNF superfamily. It is a kind of ligand, and it has various kinds of ligands such as anti-inflammatory reaction, It is known as a cytokine that regulates the response. However, there is no study on the relationship between TWEAK and autoimmune diseases.

Korea Patent Publication No. 2007-0029677

The present invention has been accomplished by confirming that the autoimmune disease can be effectively treated by suppressing the signal of TWEAK.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating an autoimmune disease comprising an TWEAK inhibitor as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating an autoimmune disease comprising an TWEAK inhibitor as an active ingredient.

In one embodiment of the present invention, the TWEAK inhibitor may be TWEAKR-Fc.

In one embodiment of the present invention, the TWEAK inhibitor may be one that inhibits the production of antibody-producing B cells or reduces the activity thereof, thereby exhibiting the therapeutic effect of an autoimmune disease.

In one embodiment of the present invention, the autoimmune disease is selected from the group consisting of rheumatoid arthritis, asthma, dermititis, psoriasis, cystic fibrosis, late organ transplantation, Multiple sclerosis, systemic lupus erythematosus, Sjogren's syndrome, Hashimoto's thyroiditis, polymyositis, scleroderma, postmenopausal osteoporosis, multiple sclerosis, inflammatory bowel disease, scleroderma, Addison disease, vitiligo, pernicious anemia, glomerulonephritis and pulmonary fibrosis, inflammatory bowel dieses, autoimmune diabetes ), Diabetic retinopathy, rhinitis, ischemia-reperfusion injury, post-angioplasty restenosis, Chronic obstructive pulmonary diseases (COPD), Graves disease, gastrointestinal allergies, conjunctivitis, atherosclerosis, coronary artery disease, angina ), Cancer metastasis, and small artery disease.

The present invention relates to a pharmaceutical composition for preventing or treating an autoimmune disease comprising an TWEAK inhibitor as an active ingredient. The TWEAK inhibitor according to the present invention can effectively treat autoimmune diseases such as arthritis and lupus by inhibiting TWEAK signaling by targeting B cells.

Figure 1 shows the results of an analysis of arthritis index and arthritis incidence according to TWEAKR-Fc treatment.
FIG. 2 shows the results of real-time PCR and confocal microscopy of the degree of B cell expression by TWEAK treatment in a lupus animal model. FIG. 2A shows the increase of B cell activation transcription factor by stimulating TWEAK by isolating CD19 + B cells from the splenocytes of Sanroque mice. FIG. 2B shows the splenocytes, CD4 + T cells and CD19 + B cells isolated from the spleen of Sanroque mice As a result of confirming the expression of Fn14, a TWEAK receptor, the expression of Fn14 was high in B cell, and Fig. 2C. Confocal microscopy revealed the expression of B220 + B cells and Fn14, a TWEAK receptor, in the spleen of Sanroque mice.
FIG. 3 shows the result of analysis of the amount of immunoglobulin in serum after injecting TWEAKR-Fc into an animal model of lupus. FIG. 3A shows IgG expression in serum of TWEAKR-Fc protein-treated mouse, FIG. 2B shows that anti-dsDNA was less in the TWEAKR-Fc group than in the case of FIG. 2A, when Anti-dsDNA was measured in the same serum.
Figure 4 shows the results of confocal microscopy of the degree of GCDML formation in spleen tissues of a group injected with TWEAKR-Fc into a lupus animal model and a control group without injection (4a) (Fig. 4b). Specifically, FIG. 4 shows the results of experiments in which sarcoque mice, a model of spontaneous lupus, were sacrificed three times a week for 3 weeks, and sacrificed. FIG. 4A shows the results of GC B cells The results of the confocal microscopy showed that the size of GC B cells was decreased in the TWEAKR-Fc protein-treated group. In the case of 4B, the cells were isolated from the spleen of the TWEAKR-Fc protein- The results showed that the expression level of the TWEAKR-Fc protein was lowered.
FIG. 5 shows the results of observing the expression level of follicular helper T (Tfh) cells in splenic tissue using a confocal microscope after immune staining in a group injected with a lupus animal model TWEAKR-Fc and a control group not injected.
FIG. 6 shows the results of confirming that glomerulonephritis induced by injection of TWEAKR-Fc can be improved. FIG. 6A shows the results of TWEAKR-Fc FIG. 6B shows that when the expression of IgG was observed in the kidney of the mouse treated with TWEAKR-Fc protein, the expression of TWEAKR-Fc protein was lowered in the group treated with TWEAKR-Fc protein .

The present invention is characterized by providing a pharmaceutical composition for the prevention or treatment of an autoimmune disease comprising an TWEAK inhibitor as an active ingredient.

TWEAK, a member of the TNF ligand family, is known to act as a proinflammatory cytokine, and the present invention has demonstrated that it is possible to treat autoimmune diseases through the modulation of TWEAK.

Wherein the TWEAK inhibitor is an antibody to a TWEAK ligand; An antibody to a TWEAK receptor; An agent that alters the binding of a TWEAK ligand to a TWEAK receptor; And agents capable of interfering with intracellular signaling of the TWEAK receptor. In one embodiment of the present invention, antibodies against TWEAK receptors were used.

The autoimmune diseases to which the composition of the present invention can be treated include, but are not limited to, rheumatoid arthritis, asthma, dermitis, psoriasis, cystic fibrosis, Post-transplantation late and chronic solid organ rejection, Multiple Sclerosis, systemic lupus erythematosus, Sjogren syndrome, Hashimoto thyroiditis, multiple myositis polymyositis, scleroderma, Addison disease, vitiligo, pernicious anemia, glomerulonephritis and pulmonary fibrosis, Inflammatory Bowel Dieses, autoimmune diseases such as autoimmune diseases, It has been reported that autoimmune diabetes, diabetic retinopathy, rhinitis, ischemia-reperfusion injury, (Eg, post-angioplasty restenosis, chronic obstructive pulmonary diseases (COPD), Graves disease, gastrointestinal allergies, conjunctivitis, atherosclerosis, coronary artery disease Coronary artery disease, angina, cancer metastasis, and small artery disease.

The term treatment refers to reversing, alleviating, inhibiting, or preventing the disease or disorder to which the term applies, or one or more symptoms of the disease or disorder, unless otherwise stated, As used herein, the term " treatment " refers to an act of treating when the treatment is defined as above. The treatment or therapies of autoimmune diseases in mammals thus may include one or more of the following:

(1) inhibiting the growth of autoimmune diseases, i.e., inhibiting its development,

(2) preventing the spread of autoimmune diseases, i.e., preventing metastasis,

(3) alleviating autoimmune diseases.

(4) preventing recurrence of autoimmune disease, and

(5) palliating symptoms of autoimmune disease

Compositions for the prevention or treatment of autoimmune diseases according to the present invention may comprise a pharmaceutically effective amount of a TWEAK inhibitor alone or may comprise one or more pharmaceutically acceptable carriers, excipients or diluents. A pharmaceutically effective amount as used herein refers to an amount sufficient to prevent, ameliorate, and treat symptoms of autoimmune diseases.

The pharmaceutically effective amount of the TWEAK inhibitor 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 pharmaceutically effective amount may be appropriately changed depending on the degree of the autoimmune disease symptoms, the age, body weight, health condition, sex, administration route and treatment period of the patient.

The term " pharmaceutically acceptable " as used herein 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 humans. 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 autoimmune disease according to the present invention may be administered through various routes including oral, transdermal, subcutaneous, intravenous, or muscular, and the dose of the active ingredient may vary depending on the administration route, , Body weight and severity of the patient, and the composition for preventing or treating autoimmune disease according to the present invention may be selected according to various factors such as known compounds having an effect of preventing, ameliorating or treating symptoms of autoimmune diseases May be administered concurrently.

Hereinafter, the present invention will be described in detail by way of the following examples, but the present invention is not limited thereto for any reason.

< Example  1>

TWEAKR - Fc Of Arthritis Treatment

The present inventors conducted the following experiments to confirm whether TWEAKR-Fc has an effect of treating arthritis. To prepare arthritis-induced mice, SKG mice were injected IP with zymosan and then injected 2 weeks later to induce an arthritis animal model. One week after the first injection of Zymosan, 50 μg of TWEAKR-Fc protein was injected three times a week for a total of 4 weeks, and then the degree of arthritis progression was observed.

As a result of the analysis, as shown in Fig. 1, the group injected with TWEAKR-Fc showed a decrease in arthritis index and a marked decrease in the incidence of arthritis compared to the group not injected.

< Example  2>

TWEAKR - Fc of Lupus  Analysis of treatment effect

<2-1> Promotion of B cell activation

CD19 + B cells were isolated from Sanroque mouse spleen, an animal model that naturally exhibits lupus disease characteristics. After stimulation of cells with TWEAK in isolated CD19 + B cells, the expression level of B-cell related transcription factors was analyzed by real-time PCR Respectively.

As shown in FIG. 2, the expression of AID, Blimp-1 and IRF4 was increased during TWEAK stimulation and mRNA expression of TWEAK receptor Fn14 in splenic CD4 + T cells and splenic CD19 + B cells of Sanroque mice Also, the increase of Fn14 expression in B220 + B cells in the spleen tissue of Sanroque mouse was confirmed by confocal microscopy.

<2-2> Loops  Analysis of disease improvement effect

Serauroque mice were injected with 100 μg of TWEAKR-Fc and control-Fc, respectively, three times per week for 3 weeks in total, after which serum was obtained from the mice and the amount of immunoglobulin in the serum was measured.

As shown in FIG. 3, the amount of immunoglobulin in the serum was decreased compared with the group not injected with TWEAKR-Fc, and the amount of anti-dsDNA was decreased compared with the control group .

Therefore, the present inventors have found that TWEAKR-Fc can improve lupus disease through inhibition of immune response.

<2-3> TWEAKR - Fc through GC  Formation decrease observation

After splenic tissue was isolated from the mouse used in the above Example <2-2>, the degree of GC formation in spleen tissue was observed through a confocal microscope. The GC is referred to as the germinal center, which, with the help of T cells in the follicle, rapidly grows and activates B cells to form a spherical structure that is histologically dyschromic, which is called the germinal center .

As a result, as shown in FIG. 4, GC formation in the spleen tissue of the TWEAKR-Fc-injected group was decreased compared with that of the control group, and mRNA expression of factors related to B-cell activity in the splenocytes of the TWEAKR- Respectively.

<2-4> TWEAKR - Fc through Tfh  Cell reduction assay

The splenic tissues of the TWEAKR-Fc injected group were isolated from the mice used in the above Example <2-2>, and the expression level of follicular helper T (Tfh) cells in the splenic tissues was analyzed by confocal microscopy after immunostaining .

As a result of the analysis, it was confirmed that the number of Tfh cells in the splenic tissue of the TWEAKR-Fc-injected group was remarkably reduced as compared with the control group, as shown in Fig.

<2-5> TWEAKR - Fc Glomeruli through Nephritis  Analysis of improvement effect

The kidney tissues of the TWEAKR-Fc injected group used in Example <2-2> were subjected to H & E staining and PAS staining method to analyze the degree of glomerulonephritis.

In addition, the amount of urinary albumin in the kidney tissue of the TWEAKR-Fc injected group was examined by tissue staining and ELISA.

As shown in FIG. 6, the degree of glomerulonephritis was decreased in the group injected with TWEAKR-Fc compared to the group injected with TWEAKR-Fc, and the amount of urinary albumin in the kidney tissue was also injected with TWEAKR-Fc Compared to the control group.

Taken together these results, the present inventors have found that TWEAKR-Fc has a function of treating autoimmune diseases such as arthritis and lupus, and that it can induce therapeutic effect by targeting B cells I could.

The present invention has been described above with reference to preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. 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.

Claims (4)

A pharmaceutical composition for preventing or treating an autoimmune disease comprising an TWEAK inhibitor as an active ingredient. The method according to claim 1,
Wherein the TWEAK inhibitor is TWEAKR-Fc. The method according to claim 1,
Wherein the TWEAK inhibitor inhibits the production of antibody-producing B cells or reduces the activity thereof, thereby exhibiting a therapeutic effect of an autoimmune disease. The method according to claim 1,
The autoimmune disease may be selected from the group consisting of rheumatoid arthritis, asthma, dermatitis, psoriasis, cystic fibrosis, post-transplantation late chronic solid organ rejection Multiple sclerosis, systemic lupus erythematosus, Sjogren's syndrome, Hashimoto thyroiditis, polymyositis, scleroderma, Addison disease, Glomerulonephritis and pulmonary fibrosis, inflammatory bowel dieses, autoimmune diabetes, diabetic retinopathy, diabetic retinopathy, diabetic retinopathy, diabetic retinopathy, diabetic retinopathy, Rhinitis, Gingival-reperfusion injury, Post-angioplasty restenosis, Chronic obstruc- tive disease (Chronic obstruc- tive) cystic fibrosis, ctive pulmonary diseases (COPD), Graves disease, gastrointestinal allergies, conjunctivitis, atherosclerosis, coronary artery disease, angina, cancer metastasis, &Lt; RTI ID = 0.0 &gt; diseases. &Lt; / RTI &gt;

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