KR20170050540A - Pharmaceutical composition for preventing or treating skin diseases comprising metformin as active ingredient - Google Patents

Abstract

The present invention relates to a composition capable of effectively treating skin disease including psoriasis or scleroderma. More specifically, the present invention relates to a pharmaceutical composition for preventing or treating skin disease including metformin as an active ingredient. According to the present invention, the composition including metformin reduces erythema, dead skin cells, and epidermal thickness, inhibits expression of IL-17, IL-6, which are inflammatory cytokines, and STAT3, alleviates damage to epidermal tissue, and, consequently, can treat psoriasis in an effective manner. In addition, the composition inhibits activity of fibrosis, inhibits expression of IL-17, IL-6, which are inflammatory cytokines, and expression of Th17 cell, inhibits COLI1a and TGF- genes, which are representative transcriptors of fibrosis, and thus can effectively treat scleroderma.

Description

[0001] The present invention relates to a pharmaceutical composition for preventing or treating skin diseases, which comprises metformin as an active ingredient,

The present invention relates to a composition capable of effectively treating skin diseases including psoriasis or scleroderma, and more particularly, to a pharmaceutical composition for preventing or treating skin diseases comprising metformin as an effective ingredient.

Psoriasis is a very common skin inflammatory disease that affects about 1 to 2% of the current population and affects about 100 million people worldwide. It is an autoimmune disease. When psoriasis spreads very little, almost all skin of the whole body is covered. Although the cause of psoriasis has not yet been fully elucidated, the Koebner phenomenon is known to be affected by genetic factors and partial psoriasis when the skin is damaged, and various environmental factors have been suggested to aggravate psoriasis . This includes stress, interruption of the permeable corticosteroids, and other environmental factors. Psoriasis is a disease that does not interfere with life, but it causes psychological pressure, loss of self-confidence, depression, and is a disease that requires long-term treatment.

In psoriasis, unlike the normal immune system, T cells are activated and secrete cytokines even when no antigen is present in the skin. These cytokines act on normal skin cells that regenerate and mature over a period of about 28 days, To reproduce and mature at a very fast 3 to 4 day cycle. This rapidly renewed and mature skin migrates to the surface of the skin, where dead cells accumulate to form grayish white streaks, blood vessels expand, blood flow increases, and skin color turns red. Regarding T cell activation in psoriasis, it has been reported that Th-1 cells trigger psoriasis and psoriasis symptoms are alleviated when IL-10 inhibits the activity of Th-1 cells. In addition, cytokines such as IL-1β, IL-6, IL-8, GM-CSF and TNF-α induce psoriasis triggering and VEGT and Substances P concentrations are also associated with psoriasis.

Common treatments for psoriasis include topical, phototherapy, and underwear medication. Topical treatments are treatments that include steroids, coal tar, anthralin, vitamin D3 and its analogs, retinoids, sunburn, etc. These topical treatments have side effects of skin thinning, stretch marks, burns, irritation and photosensitivity. Phototherapy involves the administration of the sunralen with ultraviolet B or ultraviolet A and has the disadvantage that skin ages quickly and the incidence of skin cancer is increased. The topical application to the most severe psoriasis includes administration of methotrexate, oral retinoid, cyclosporine, and the like. However, since cyclosporine may cause kidney damage and blood pressure problems, careful observation is needed, and methotrexate is effective for both psoriasis and psoriatic arthritis. However, cumulative use of methotrexate may cause cirrhosis or fibrosis. Should be used. In addition, the use of oral retinoids should be used with caution because they can seriously impair women's fertility. Thus, drugs conventionally used to treat psoriasis have side effects or disadvantages, and drugs such as methotrexate and cyclosporin are drugs that inhibit the activity of T cells and are so toxic that they have less side effects than these drugs Drugs such as Amevive, Enbrel, Inficiximab and Raptiva have been developed as biologics.

Skin sclerosis is also referred to as systemic sclerosis, and systemic fibrosis forms include two subtypes depending on the degree of skin involvement. Restricted skin types affect limbs, while decentralized subtypes also affect the body. In addition to systemic sclerosis, there is also a morphea that does not affect the internal organs. Systemic sclerosis is a widespread microvascular injury and progressive fibrosis in the skin and internal organs, and generally the most obvious clinical manifestations are, for example, skin hardening, flushing, It is a related scar. Currently, some available agents can soften the skin and reduce inflammation, but additional and effective drugs are needed for the pathogenesis of scleroderma.

Metformin has been used in the treatment of non-insulin dependent diabetes mellitus (NIDDM) as an oral antidiabetic agent and as a blood-sugar drug in the treatment of NIDDM patients by lowering both basal plasma glucose and postprandial plasma glucose Improves glucose tolerance. Metformin has been widely used as a treatment for type 2 diabetes, and clinical approaches for polycystic ovary syndrome, weight loss, and cancer treatment have been made. However, the use of metformin for the prevention and treatment of skin diseases has not been disclosed until now.

Korea Patent Publication No. 2009-0005513

Accordingly, the present inventors have completed the present invention by confirming that metformin is effective in suppressing psoriasis and scleroderma by reducing erythema, keratin and epidermal thickness, alleviating damage to skin epidermal tissue, and inhibiting fibrosis activity.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating skin diseases comprising metformin as an active ingredient.

Another object of the present invention is to provide a method for treating skin diseases comprising administering metformin to a subject suffering from a skin disease other than human.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating skin diseases comprising metformin as an active ingredient.

Further, in one embodiment of the present invention, the composition may contain metformin in an amount of 0.05 to 20 mg.

Further, in one embodiment of the present invention, the skin disease may be selected from psoriasis or scleroderma.

Also, in one embodiment of the present invention, the composition may reduce or inhibit the expression of IL-17, IL-6 or STAT3, which is an inflammatory cytokine.

In one embodiment of the present invention, the composition may inhibit the expression of fibroblast transcription factors COLIla and TGF-beta.

The composition comprising metformin according to the present invention reduces erythema, keratin and epidermal thickness and inhibits the expression of the inflammatory cytokines IL-17, IL-6 and STAT3, thereby alleviating damage to skin epidermal tissue and, consequently, Can be effectively treated. In addition, the composition inhibits the activity of fibrosis, inhibits the expression of inflammatory cytokines IL-6 and STAT3 and the expression of Th17 cells, inhibits the expression of COLI1a and TGF-beta genes, which are typical transcription factors of fibrosis, Can effectively treat sclerosis.

FIG. 1A is a Psoriasis Area and Severity Index (PASI) score that measures the clinical severity of psoriasis in normal control (NC), vehicle control (VC), and experimental group (metformin) And the results are shown in FIG. Figure 1B is a graph showing PASI scores in normal, control, and experimental groups.
Figure 2 shows the results of hematoxylin and eosin (H & E) staining of normal, control and experimental skin tissues.
FIG. 3 shows the results of comparing the expression of IL-17, IL-6 and STAT3 in the normal group, the control group and the experimental group.
FIG. 4 shows the visual acuity of skin sclerosis improvement in the normal group, the control group and the experimental group.
Figure 5 shows the results of hematoxylin and eosin (H & E) staining of lung tissues in normal, control and experimental groups.
6 shows the results of comparing the thickness of the dermis and the number of hair follicles in the normal group, the control group and the experimental group.
Figure 7 shows the results of comparing the expression of IL-6 and STAT3 in normal, control and experimental groups.
FIG. 8 shows the results of comparing the amount of expression of Th17 cells after staining anti-CD4-pe and anti-IL-17-FITC in cells isolated from the spleen of normal, control and experimental groups.
FIG. 9 shows the results of analysis of the expression of COLI1a and TGF-beta gene by real-time PCR by isolating RNA by treating RNA zolB to cells isolated from the skin of control and experimental groups, and then synthesizing cDNA from the isolated RNA.

The present invention relates to a pharmaceutical composition for preventing or treating skin diseases comprising metformin as an active ingredient.

The present inventors have focused on the metformin compounds of the present invention while studying new drugs capable of preventing or treating skin diseases, and metformin has been known to be effective for the treatment of diabetes. Korean Patent Laid-Open Publication No. 2009-0005513 discloses that metformin malonate has antidiabetic activity, but there is no mention of the use of metformin for the prevention and treatment of skin diseases.

The usual treatments for psoriasis include topical medication, phototherapy, and underwear medication, but topical treatment has side effects such as skin thinning, stretch marks, burns, irritation, and photosensitivity, while phototherapy is administered with ultraviolet B or ultraviolet A And the skin is rapidly aged and the incidence of skin cancer is increased. Thus, drugs conventionally used to treat psoriasis have side effects or disadvantages, and drugs such as methotrexate and cyclosporin are drugs that inhibit the activity of T cells and are highly toxic. Therefore, And the like. In addition, in order to treat scleroderma, there is currently used an agent that softens the skin and reduces inflammation. However, there is a need for a drug that can fundamentally treat scleroderma.

Hereinafter, a pharmaceutical composition for preventing or treating skin diseases comprising metformin as an active ingredient according to an embodiment of the present invention will be described in detail. This example focused on experiments on psoriasis and skin sclerosis models. However, the present invention is not limited thereto and can be applied to all skin diseases.

According to one embodiment of the present invention, it has been found that the composition comprising metformin reduces erythema, keratin, epidermal thickness.

According to one embodiment of the present invention, it has been found that a composition comprising metformin reduces or inhibits the expression of IL-17, IL-6 or STAT3, an inflammatory cytokine.

According to one embodiment of the present invention, it has been found that the composition containing metformin inhibits the expression of fibroblast transcription factors, COLIla and TGF-beta genes.

The metformin contained in the composition of the present invention may be used in the form of the respective compound itself, preferably in the form of 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 compound according to the present invention can be isolated from nature or can be prepared by chemical synthesis methods known in the art. The metformin compound according to the present invention can be isolated from nature or can be prepared by chemical synthesis methods known in the art.

In addition, the target disease to which the composition according to the present invention can be treated may be a skin disease. In the present invention, the " skin " means all skin including mammals hair, nails and toenails including humans. In addition, " skin disease " used in the present invention means all abnormalities appearing in the " skin ".

Examples of the skin diseases that can be applied in the present invention include, but are not limited to, psoriasis, skin sclerosis, larynx, bowen's disease, papilloma, eczema, addictive eczema, atopic dermatitis, , Seborrheic dermatitis, squamous cell carcinoma, squamous cell carcinoma, foot pediculosis, biliary keratosis, ray keratosis, and the like.

Also, in the present invention, the treatment means, unless otherwise stated, a disease or condition to which the term applies, or a condition that reverses, alleviates, inhibits or prevents the progression of one or more symptoms of the disease or condition And the term " treatment " as used herein refers to an act of treating when defined as above.

The composition for the prevention or treatment of skin diseases according to the present invention may comprise a pharmaceutically effective amount of metformin or may comprise one or more pharmaceutically acceptable carriers, excipients or diluents. The pharmacologically effective amount is an amount sufficient to prevent, ameliorate, and treat the symptoms of a skin disease, and is appropriately determined depending on the severity of the skin disease, the age, body weight, health condition, sex, Can be changed.

Also, the pharmaceutically acceptable hereinabove is physiologically acceptable and refers to a composition which, when administered to a human, does not normally cause an allergic reaction such as a gastrointestinal disorder, dizziness, or the like. 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 skin diseases 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 route of administration, The composition of the present invention can be appropriately selected according to various factors such as body weight and the severity of the patient and the composition for preventing or treating skin diseases according to the present invention can be used in combination with known compounds having the effect of preventing, Lt; / RTI >

Accordingly, the present invention can provide a medicament for preventing or treating skin diseases, wherein a composition comprising metformin as an active ingredient is provided.

Further, the present invention can provide a method for treating skin diseases, which comprises administering metformin to a subject suffering from a skin disease other than human, in combination.

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  One] Metformin  Depending on the treatment psoriasis  Measure improvement and Psoriatic  Clinical severity degree analysis

To examine whether metformin is effective for the prevention and treatment of psoriasis, the present inventors prepared a psoriasis model by applying imiquimod 5% cream to C57BL / 6 normal mice, and administered orally administered metformin to the animal model, Were measured.

For this purpose, we used a C57BL / 6 normal mouse, such as 8-10 weeks old, and a hairless epilating cream (Veet ^® , Korea) on the hips, and after 1 day, Aldara ^®, 3M Health care Limited, UK) was applied at 62.5 mg / mice every day for 6 days. In the animal model, metformin (10 mg / kg) was administered orally from the second day after psoriasis induction to determine the degree of improvement of psoriasis visually and to measure the clinical severity of psoriasis. PASI score (Psoriasis Area and Severity Index) And thickness. The experimental group was the group treated with metformin and the control group was treated with physiological saline dissolved in metformin instead of metformin.

As a result, from the day after the application of quim mode, the control group showed a significant clinical change of psoriasis due to an increase in erythema, keratinization and thickening of skin lesions. On the other hand, the clinical severity of erythema, keratinization, and thickening was observed in the experimental group, but gradually increased compared to the control group, and the severity of clinical severity was significantly decreased from 4 to 5 days. (See Figs. 1A and 1B).

[ Example  2] psoriasis  In animal models Metformin  Analysis of Skin Damage in Skin Tissue by Treatment

The present inventors conducted a histological examination of the skin tissue of an animal model of psoriasis-induced mice in order to analyze the degree of improvement of epidermal tissue damage in skin tissue following treatment with metformin.

For this purpose, the skin of each mouse group was fixed with 10% formalin and the block was made with paraffin on the 8th day after induction of psoriasis in the experimental group and the control group. Skin sections were prepared and attached to the slide. After deparaffinization with xylene, the skin sections were dyed with high concentration and low concentration of ethanol, and skin sections were stained with hematoxylin and eosin.

As a result of observation of the stratum corneum layer thickness and inflammation level in the experimental group after the experiment, it was found that the stratum corneum and granule layer of the control group without treatment with metformin were significantly thickened and damaged, , And visual and numerical confirmation that the degree of damage was alleviated (see FIG. 2).

[Example 3] Inhibitory effect of epidermal inflammatory cytokine expression in skin tissue by metformin treatment in an animal model of psoriasis

The present inventors further performed a histological examination of the skin tissue of the psoriasis-induced mouse animal model to analyze the expression of inflammatory cytokines in the epidermis of the skin tissue following metformin treatment.

To this end, the present inventors prepared skin slices in the same manner as in Example 2, attached to slides, and then immunohistochemically stained IL-17, IL-6 and STAT3, typical inflammatory cytokines related to autoimmune psoriasis And analyzed with an optical microscope.

As a result, the expressions of IL-17, IL-6 and STAT3 were significantly inhibited in the skin tissue of the metformin-treated group of the present invention compared with the control group. From the results, it was found that metformin inhibited the activity of inflammatory cytokines in autoimmune psoriasis (See FIG. 3).

That is, according to the above Examples 1 to 3, metformin reduces erythema, keratin, epidermal thickness, and inhibits the expression of inflammatory cytokines IL-17, IL-6 and STAT3, And the effect of suppressing psoriasis was excellent. Thus, it can be seen that the composition comprising metformin of the present invention can be effectively used for prevention or treatment of psoriasis.

[ Example  3] psoriasis  In animal models Metformin  Of Inhibitory Effect on Skin Epidermal Inflammatory Cytokine Expression by Treatment

The present inventors further performed a histological examination of the skin tissue of the psoriasis-induced mouse animal model to analyze the expression of inflammatory cytokines in the epidermis of the skin tissue following metformin treatment.

To this end, the present inventors prepared skin slices in the same manner as in Example 2, attached to slides, and then immunohistochemically stained IL-17, IL-6 and STAT3, typical inflammatory cytokines related to autoimmune psoriasis And analyzed with an optical microscope.

As a result, the expressions of IL-17, IL-6 and STAT3 were significantly inhibited in the skin tissue of the metformin-treated group of the present invention compared with the control group. From the results, it was found that metformin inhibited the activity of inflammatory cytokines in autoimmune psoriasis (See FIG. 3).

That is, according to the above Examples 1 to 3, metformin reduces erythema, keratin, epidermal thickness, and inhibits the expression of inflammatory cytokines IL-17, IL-6 and STAT3, And the effect of suppressing psoriasis was excellent. Thus, it can be seen that the composition comprising metformin of the present invention can be effectively used for prevention or treatment of psoriasis.

[ Example  4] Metformin  Measuring degree of improvement of skin sclerosis according to treatment and analysis of degree of severity

In order to examine whether metformin is effective in preventing and treating skin sclerosis, the present inventors firstly dissected C57BL / 6 or balb / c normal mouse hair and then prepared bleomycin was dissolved in 0.9% sodium chloride (1 mg / ml), 100 ul each day was subcutaneously injected subcutaneously from the throat to the back of the neck for 4 weeks, and the sclerosis index was visually measured by applying metformin to the animal model .

To this end, the present inventors used a hair removal cream (Veet ^® Korean product) to epilate from the neck of a C57BL / 6 or balb / c normal mouse at 8-10 weeks of age, and after 1 day, the bleomycin was dissolved in 0.9% sodium chloride (1 mg / ml), 100 ul each day for 4 weeks, and subcutaneously injected into each of four corners of the square at the back of the neck. In the animal model, metformin (10 mg / kg) was applied to the skin one week after the onset of sclerosis, and the degree of improvement of sclerosis was visually measured and compared with the severity of skin thickness. The experimental group was the group treated with metformin and the control group was treated with physiological saline dissolved in metformin instead of metformin.

As a result, from the first week of application of bleomycin subcutaneous injection, thickening of the skin was increased in the control group, and it was visually confirmed that hair did not grow remarkably. In the experimental group, hairiness was significantly prolonged compared to the control group, and skin thickness similar to that of the normal skin was visually confirmed (see FIG. 4).

[ Example  5] Skin sclerosis in animal models Metformin  Analysis of damage caused by fibrosis of lung tissue by treatment

The present inventors conducted a histological examination of the lung tissue of a sclerosis-induced mouse animal model in order to analyze the degree of improvement of lung tissue damage by metformin treatment.

For this purpose, the mice to which metformin was administered and the control group were fixed with 10% formalin in each mouse group on day 28 after induction of sclerosis, and blocks were prepared with paraffin. From these, the lung sections were prepared and adhered to the slides. After deparaffinization with xylene, high density and low density ethanol were used, and the lung sections were stained with hematoxylin and eosin.

As a result of observing the pulmonary fibrosis of the experimental group after the above experiment, the lung fibrosis of the control group without methampin treatment was remarkably increased, but the experimental group showed a decrease in pulmonary fibrosis compared to the control group, Histology and severity index (see Figure 5).

[ Example  6] In an animal model of scleroderma Metformin  Analysis of dermal damage in skin tissue by treatment

The present inventors conducted a histological examination of the skin tissue of a sclerosis-induced mouse animal model in order to analyze the degree of improvement of dermal damage in the dermal tissue following metformin treatment.

For this purpose, the animals to which metformin was administered and the control group were fixed with 10% formalin in each mouse group on day 28 after induction of sclerosis, and blocks were prepared with paraffin. Skin sections were prepared and attached to the slide. After deparaffinization with xylene, the skin sections were dyed with high concentration and low concentration of ethanol, and skin sections were stained with hematoxylin and eosin. Skin sections were also stained with Masson's Trichrome (MT) to determine the density of the collagen fibers.

As a result of observing the thickness of the skin dermis and the density of the collagen fibers in the test group after the experiment as described above, it was found that the density of the collagen fibers in the dermis and the density of the collagen fibers in the control group without metformin was dense, (Fig. 6). However, in the experimental group, the decrease in dermal thickness or the increase in the number of hair follicles compared to the control group was confirmed to be mitigated to the same phenomenon as the normal group.

[ Example  7] In skin models of sclerosis animals Metformin  Of Inhibitory Effect of Dermal Inflammatory Cytokine Expression in Skin Tissue by Treatment

The present inventors further performed histological examination of the skin tissue of the skin sclerosis-induced mouse animal model to analyze the expression of inflammatory cytokines in the dermis of the skin tissue following metformin treatment.

To this end, the present inventors prepared skin slices in the same manner as in Example 3, adhered to slides, stained IL-6 and STAT3, which are typical inflammatory cytokines related to autoimmune sclerosis, using immunochemical staining, This was analyzed with an optical microscope.

As a result, the expression of IL-6 and STAT3 in the skin tissue of the group treated with metformin of the present invention was markedly suppressed as compared with that of the control. From the results, metformin could control the activity of inflammatory cytokines in autoimmune scleroderma (See Fig. 7).

[ Example  8] In an animal model of skin sclerosis Metformin  By treatment Th17  Analysis of inhibition effect on cell expression

In order to investigate the effect of the metformin treatment of the present invention on Th17 cells, the present inventors stained anti-CD4-pe and anti-IL-17-FITC cells in a mouse spleen, Were confirmed using a confocal microscope.

As a result, the expression of Th17 cells, which is an apoptotic cell, was significantly suppressed in the spleen of mice treated with metformin, compared with the control group, skin sclerosis experimental animals. From the above results, it was confirmed that metformin can inhibit the expression and activity of Th17 cells (see FIG. 8).

[ Example  9] In scleroderma animal models Metformin  Analysis of inhibition of fibrosis in skin cells by treatment

In order to investigate the effect of the metformin treatment of the present invention on skin cells, the present inventors treated RNA zolB to cells isolated from sclered skin of mice to isolate RNA, synthesized cDNA from isolated RNA, -β gene was analyzed by real-time PCR.

As a result, the expressions of COLI1a and TGF-beta genes, which are representative transcription factors of fibrosis, in the skin of mice treated with metformin were markedly reduced as compared with control animals of skin sclerosis (see FIG. 9).

That is, according to the above Examples 4 to 9, metformin inhibits the activity of fibrosis, inhibits the expression of inflammatory cytokines IL-6 and STAT3 and the expression of Th17 cells, and the representative transcription factors of fibrosis, COLI1a and By inhibiting the expression of the TGF-beta gene, fibrosis of skin tissue was alleviated and the effect of inhibiting sclerosis was excellent. Accordingly, it can be seen that the composition containing metformin of the present invention can be effectively used for the prevention or treatment of skin sclerosis.

Claims (8)

A pharmaceutical composition for preventing or treating skin diseases comprising metformin as an active ingredient. The method according to claim 1,
Wherein the composition comprises metformin in an amount of 0.05 to 20 mg. The method according to claim 1,
Wherein the skin disease is psoriasis or scleroderma. ≪ RTI ID = 0.0 > 11. < / RTI > The method according to claim 1,
Wherein said composition reduces or inhibits the expression of IL-17, IL-6 or STAT3 which is an inflammatory cytokine. The method according to claim 1,
Wherein said composition inhibits the expression of fibroblast transcription factors COLI1a and TGF-beta gene. A method for treating skin diseases comprising administering metformin to a subject suffering from a skin disease other than human. The method according to claim 6,
Wherein the composition comprises metformin in an amount of 0.05 to 20 mg. The method according to claim 6, wherein the skin disease is psoriasis or scleroderma.

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