KR101013061B1 - An alcohol extract of Cordyceps bassiana for treating and preventing atopy dermatitis and pruritic skin lesion - Google Patents

Abstract

The present invention relates to the use and treatment of skin inflammation, including pruritus and atopic dermatitis of yellow bunch Cordyceps alcohol extract, wherein the extract of yellow bunch Cordyceps sinensis with ethanol, propanol or butanol significantly reduces the thickness of the inflamed epidermal layer, and It can reduce the IgE concentration, lower the dermatitis score and apply it as a composition for preventing and treating skin inflammation.

Skin pruritus, atopic dermatitis, yellow cordyceps, alcohol extract, butanol extract

Description

An alcohol extract of Cordyceps bassiana for treating and preventing atopy dermatitis and pruritic skin lesions

The present invention relates to the treatment and prevention of skin inflammation, including skin pruritus, atopic dermatitis, of the alcoholic extract of yellow bunch Cordyceps sinensis.

Atopic dermatitis is a chronic skin disease accompanied by dry skin, keratosis and itching. It affects 0.5% to 1% of the population, especially 5% to 10% of children, and the number of patients is increasing. Infections with atopic dermatitis significantly increase infiltration of immune-related cells such as macrophages, mast cells and Th lymphocytes. In patients with atopic dermatitis, blood IgE levels increase significantly because the number of Th2 cells increases and IL-4 secreted by these cells stimulates B lymphocytes to promote IgE secretion. Therefore, atopic dermatitis is classified as a Th2-type skin disease associated with abnormalities of the immune system. Atopic dermatitis inflicts a lot of economic damage along with physical and mental pain for the patient, but no specific drug has yet been found to treat the disease. Currently, steroids, antihistamines, etc. are used as atopic dermatitis treatments, but long-term use of these drugs causes serious side effects.

Cordyceps sinensis is a fungal insect parasite belonging to Cordyceps , and more than 300 species are known to be distributed all over the world. Cordyceps sinensis invade insect larvae, chrysalis, adult insects, etc., and kills insects, forming a fruiting body and fruit as a host. Cordyceps sinensis has long been known as a elixir of immortality in China and is widely used as a herbal medicine for the treatment of respiratory diseases including asthma and liver diseases including jaundice. In recent years, research on the enhancement of immunity of Cordyceps sinensis has been extensively conducted. There are Cordyceps bat moth (C. sinensis) extract, Cordyceps militaris reported that also serves to extract or only serve as a main component coordination sepin (cordycepin) The immunoregulatory material of Cordyceps as anti-inflammatory substances (C. militaris).

C. bassiana ( C. bassiana ), a type of cordyceps, is the first parasitic fungus to be found in the genus Butterfly Lepidoptera. Beauveria bassiana , a form that emerges during the life cycle of yellow bunched cordyceps, has been used for the treatment of neurological diseases, cancers, skin infections and wounds for a long time in China and Korea. Nothing is known about its efficacy.

Korean Patent Laid-Open Publication No. 10-2006-92373 discloses a herbal composition for the prevention and improvement of atopic dermatitis, which contains a herbal extract consisting of pollen, white nymph, Cordyceps sinensis, casualties. However, the above technique can be used for Cordyceps militaris ), as well as the oral administration of the extract of the hydrothermal extract of a number of the herbal medicines can not be said that the effect is entirely due to Cordyceps sinensis. In addition, although the above description is not toxic, oral administration of the combined herbal medicine always carries the risk of liver toxicity.

The present invention is to provide a composition that can prevent and treat dermatitis, such as atopic dermatitis or pruritus, using a natural product having no side effects.

In the present invention, a composition for inhibiting atopic dermatitis was prepared by using a yellow bunched cordyceps extract, including the fruiting body extract of the yellow bunched cordyceps. NC / Nga mice were used as model animals for atopic dermatitis, and atopic dermatitis was induced using 2,4-DNFB (hereinafter referred to as "DNFB"). This mouse not only has a naturally occurring characteristic of atopic dermatitis when raised in a normal environment, but also has a characteristic of enhancing atopic dermatitis by repeatedly applying a chemical antigen such as DNFB to the skin.

The atopic dermatitis animal model was used to induce atopic dermatitis for 4 weeks and to apply the butanol fraction (CBBF) of yellow cordyceps to petroleum jelly at different concentrations and apply it to the affected area, resulting in a concentration-dependent decrease in dermatitis score, epidermal thickness and serum IgE concentration. In addition, the acute dermatitis caused by 6 weeks was significantly improved.

Cordyceps cordyceps of the present invention alcohol extracts of bassiana ) and compositions comprising the same have been shown to lower dermatitis scores of skin pruritus and atopic dermatitis, significantly reduce the thickness of inflamed epidermal layers, and reduce serum IgE concentrations. And / or for the purpose of preventing, treating or ameliorating atopic dermatitis.

The present invention provides an alcohol extract of Cordyceps bassiana for preventing and treating skin pruritus and atopic dermatitis.

The present invention the alcohol is ethanol, propanol, and the skin pruritus, it characterized in that the at least one alcohol selected from butanol, and the prevention and treatment of atopic dermatitis yellow bundle Cordyceps (Cordyceps for bassiana ) provides an alcoholic extract. The alcohol is 5 to 100% concentration, preferably 70% concentration is used.

The present invention is a yellow bunch for skin pruritus and atopic dermatitis, characterized in that the alcohol extract is a butanol fraction obtained by adding water: butanol mixed solvent to the water fraction obtained by extracting the ethanol extract with a water: hexane mixed solvent. Provide an alcoholic extract of Cordyceps bassiana .

The present invention yellow cordyceps cordyceps Provided is a composition for preventing and treating skin pruritus and atopic dermatitis containing an alcoholic extract of bassiana ).

The invention Yellow bundle Cordyceps (Cordyceps, characterized in that the alcohols ethanol, propanol and the at least one alcohol selected from butanol Provided is a composition for preventing and treating skin pruritus and atopic dermatitis containing an alcoholic extract of bassiana ).

In the present invention, the alcohol extract is a butanol fraction obtained by adding water: butanol mixed solvent to the water fraction obtained by adding water: hexane mixed solvent to the ethanol extract, Cordyceps Provided is a composition for preventing and treating skin pruritus and atopic dermatitis containing an alcoholic extract of bassiana ).

The present invention is characterized in that the composition for the prevention and treatment of skin pruritus and atopic dermatitis is a composition for external application of the skin, yellow bunch Cordyceps Provided is a composition for preventing and treating skin pruritus and atopic dermatitis containing an alcoholic extract of bassiana ).

The present invention yellow cordyceps cordyceps Provided is a cosmetic composition for improving skin pruritus and atopic dermatitis containing an alcohol extract of bassiana ).

The invention Yellow bundle Cordyceps (Cordyceps, characterized in that the alcohols ethanol, propanol and the at least one alcohol selected from butanol Provided is a cosmetic composition for improving skin pruritus and atopic dermatitis containing an alcohol extract of bassiana ).

In the present invention, the alcohol extract is a butanol fraction obtained by adding water: butanol mixed solvent to the water fraction obtained by adding water: hexane mixed solvent to the ethanol extract, Cordyceps Provided is a cosmetic composition for improving skin pruritus and atopic dermatitis containing an alcohol extract of bassiana ).

In the present invention, NC / Nga mice, a model animal of atopic dermatitis, were applied in a normal environment for 4 weeks with DNFB applied at a concentration of 0.2% (w / v). As a result, typical changes associated with atopic dermatitis were observed, such as an increase in dermatitis scores including itching, erythema, bleeding, and peeling, thickening of the epidermal layer of the affected area, and a significant increase in serum IgE concentration.

Thus induced atopic dermatitis for 4 weeks and the yellow bunch Cordyceps butanol fraction (CBBF) obtained by the method of FIG. 1 was mixed with petrolatum at a concentration of 5, 10 or 20 mg / g and applied to the affected area for 12 days dermatitis score, Epidermal thickness and serum IgE concentrations were reduced in a concentration dependent manner. These results indicate that the alcoholic extract of the Yellow Cordyceps sinensis has an inhibitory effect on experimentally induced atopic dermatitis in experimental animals. Thus, the present inventors attempted to investigate the effect of the yellow bund Cordyceps alcohol extract according to the severity of atopic dermatitis. Extending the induction period of atopic dermatitis to 6 weeks significantly increased all the indicators such as dermatitis score, epidermal thickness, serum IgE concentration and serum histamine concentration for 4 weeks. In this state, when the butanol fraction (CBBF) of FIG. 1 was applied at a concentration of 10 mg / g, all indicators were significantly decreased, and the reduction rate of each indicator was greater than that of atopic dermatitis for 4 weeks. These results suggest that atopic dermatitis is severely induced to have a greater effect of improving butanol fraction.

On the other hand, in order to observe the effect of the common extract of yellow worm cordyceps on atopic dermatitis, atopic dermatitis was induced for 4 weeks, and the water extract and ethanol extract of the yellow worm cordyceps obtained as shown in FIG. 1 were mixed at concentrations of 10 mg / g, respectively. It was applied to the affected area for 12 days. As a result, the dermatitis score, epidermal thickness, and serum IgE concentration which were elevated by atopic dermatitis were not changed by water extract (CBWE) but decreased by ethanol extract (CBEE). These results suggest that the atopic dermatitis inhibitors contained in the yellow bunched Cordyceps sinensis are not well extracted by boiling water but are extracted by alcohols such as ethanol, propanol and butanol. Propanol extracts also showed similar results as ethanol extracts (results not shown).

Since the causes and processes of atopic dermatitis are very complicated, the results of this experiment alone cannot fully explain the mechanisms by which the extract of yellow cordyceps can improve the atopic dermatitis. However, atopic dermatitis is an immunoreactive disease and various immune cells and cytokines. This is involved. That is, dendritic cells ingesting allergens that infiltrate the affected area of atopic dermatitis differentiate CD4 ⁺ T cells into Th1 cells or Th2 cells. In general, Th1 cells secrete INF-γ to cause a delayed immune response, and Th2 cells secrete IL-4 to cause an acute immune response. IL-4 secreted from Th2 cells acts on B cells to induce Ig homologous conversion to promote IgE production, and IgE stimulates mast cells to increase histamine secretion. Therefore, alcohol extracts, especially butanol fraction (CBBF), decreased the serum IgG and serum histamine concentrations associated with atopic dermatitis, and the alcohol extracts of yellow cordyceps were inhibiting Th2 immune response to improve and prevent atopic dermatitis. It is thought to suggest that there is a function. These results indicate that the alcoholic extract of the Yellow Cordyceps sinensis shows a function of improving skin inflammation such as skin pruritus or atopic dermatitis. Therefore, the alcohol extract of the yellow bunch Cordyceps sinensis suggests a variety of applications in the treatment and prevention of skin inflammation.

A pharmaceutical composition containing an alcoholic extract of yellow bundled Cordyceps sinensis as an active ingredient may be formulated in an injectable form or a coating by conventional methods in combination with a carrier that is conventionally acceptable in the pharmaceutical art. Injectable compositions are preferably aqueous isotonic solutions or suspensions, and the compositions mentioned are sterile and / or contain auxiliaries (eg, preservatives, stabilizers, wetting or emulsifier solution promoters, salts or buffers for controlling osmotic pressure). In addition, they may contain other therapeutically valuable substances.

The pharmaceutical preparations thus prepared may be parenterally, ie, subcutaneously, intramuscularly, or topically, as desired. The dosage may range from 1 mg to 10 mg / kg in a daily dosage of 1 to 10 mg / kg. It can be divided into several doses. Dosage levels for a particular patient may vary depending on the patient's weight, age, sex, health condition, time of administration, method of administration, severity of the disease, and the like.

In addition, the coating agent containing the fusion protein of the present invention as an active ingredient can be easily prepared in any form according to a conventional manufacturing method. As an example, in the preparation of a cream coating agent, an alcoholic extract of the yellow bundle Cordyceps sinensis of the present invention is contained in a creamy oil-in-water (O / W) or water-in-oil (W / O) cream base. Antioxidants, preservatives, etc. may be used as necessary, and synthetic or natural materials such as proteins, minerals, vitamins, etc. may be used in combination for the purpose of improving physical properties.

In addition, the alcohol extract of the yellow bunch Cordyceps sinensis of the present invention can be used as a cosmetic, by adding a pH adjuster, flavoring, emulsifiers, preservatives, etc. as needed, in the usual cosmetic preparation method, such as lotion, gel, water-soluble powder, fat-soluble powder, It may be formulated with a water soluble liquid, lotion, cream, essence, shampoo, cleanser, rinse, hair conditioner and the like.

The present inventors were able to confirm that the thickness of the outer skin layer was reduced as a result of applying the alcoholic extract of the yellow bundled Cordyceps sinensis to the skin induced by atopic dermatitis in the mouse. Accordingly, it has been found that the alcoholic extract of the yellow bundled Cordyceps sinensis can be used as the main component of the pharmaceutical composition and / or coating agent (used herein in the same sense as "skin external preparation") composition.

Hereinafter, the configuration of the present invention in more detail through the embodiment. However, the scope of the present invention is not limited to the scope of the following examples.

<Materials and Methods>

< Example  1: Extraction of Biologically Active Compounds from Yellow Bunny Cordyceps Sinensis>

Yellow bunch Cordyceps sinensis: In this experiment, yellow bunch Cordyceps sinus cultivated in Mushtek (Chuncheon, Korea) was dried and used as a sample.

Water extraction and ethanol extraction: tertiary distilled water or 100% ethanol equivalent to 10 times the sample weight is mixed with the sample, heated and extracted for 4 hours, and then the extract is concentrated and lyophilized, respectively, to extract water (CBWE) and ethanol extract ( CBEE).

Preparation of butanol fraction: A dried yellow bunch of Cordyceps sinensis as a sample and 100% ethanol corresponding to 20 times the weight of the sample were mixed and extracted by heating for 4 hours, and the extract was concentrated under reduced pressure and lyophilized. The dried ethanol extract was dissolved in 0.5 L of distilled water, the same amount of hexane was mixed, and the hexane layer was separated and removed. An equal amount of 70% n-butanol was mixed with the remaining solution, and the butanol layer was separated, concentrated, and lyophilized to obtain a butanol fraction (CBBF) (FIG. 1).

< Example  2: Observation of the healing effect of each extract of yellow Cordyceps sinensis against atopic dermatitis>

Induction of Atopic Dermatitis in Experimental Animals: NC / Nga mice (male, 6 weeks old), a model animal of atopic dermatitis, were used in this study. The animals were used for experiments after adapting the animals for 1 week in a normal breeding room where the temperature and humidity were maintained at 20 ° C. and 50%, and the contrast was changed every 12 hours. The back of the NC / Nga mouse was epilated and left for 24 hours to heal fine wounds on the skin. 150 μl of acetone / olive oil (3: 1) containing 0.2% 2,4-DNFB (dinitrofluorobenzene) was applied twice a week for 4 weeks to induce atopic dermatitis. Extended to 6 weeks.

Treatment of Various Extracts of Yellow Cordyceps sinensis for Atopic Dermatitis: In the normal control group, only the back area was epilated, and in the atopic control group, dermatitis was induced and only petrolatum was applied. In the experimental group, atopic dermatitis was induced, and then water extract (CBWE), ethanol extract (CBEE), and butanol fraction (CBBF) mixed with petroleum jelly were applied twice daily to the affected area for 12 days.

< Example  3: dermatitis score ( dermatitis score Output)

After the application of each extract of Cordyceps sinensis, the affected area was observed, and gross changes such as erythema, bleeding, and scab were evaluated in 1 ~ 10 stages and used as dermatitis scores.

< Example  4: Epidermal thickness measurement>

In the center of the affected area, skin tissue was collected in a size of 1 cm × 1 cm, fixed in 10% formaldehyde solution, embedded in paraffin, and a section having a thickness of 4 μm was prepared. Tissue sections were stained with hematoxylin-eosin and then magnified 200 times using an optical microscope and the thickness of the epidermal layer was measured by selecting 5 fields of view.

< Example  5: serum IgE  Concentration and Histamine Concentration>

Blood was collected from the heart and serum IgE and histamine levels were measured by ELISA (Enzyme-linked immunosorbent assay).

Statistical processing of measured values

All measurements obtained in this experiment are presented as mean ± standard deviation. The mean difference was statistically tested using Student's t test and the significance level was less than 5%.

Experiment result

1. Water extract against atopic dermatitis ( CBWF ) And ethanol extract ( CBEF ) Effect

Atopic dermatitis was induced over 4 weeks, and water extract (CBWE) and ethanol extract (CBEE) of yellow bunched cordyceps were applied to the affected area for 12 days at a concentration of 10 mg / g, respectively. As a result, the dermatitis score, epidermal thickness, and serum IgE concentration increased by atopic dermatitis were not affected by water extract, but decreased by ethanol extract (Table 1).

Table 1 shows the effects of water and ethanol extracts on the indices of increased dermatitis caused by atopic dermatitis. DNFB was applied for 4 weeks to induce atopic dermatitis, and each extract was mixed with petrolatum at a concentration of 10 mg / g. This is the result obtained by coating for 12 days. Experimental Value: 5 Means ± Standard Error.

Atopy control
Water Extract (CBWE)
Ethanol Extract (CBEE)
Dermatitis score
7.33 ± 0.67
8.67 ± 1.20
6.67 ± 0.33
Skin layer thickness (㎛)
236.42 ± 28.94
217.48 ± 45.92
195.28 ± 9.35
Serum IgE Concentration (μg / mL)
10.86 ± 2.24
13.77 ± 1.81
7.86 ± 3.06

2. against atopic dermatitis Butanol  Fraction CBBF Concentration-dependent effects of

Atopic dermatitis was induced over 4 weeks and the butanol fraction (CBBF) was applied to the affected area for 12 days at a concentration of 5, 10 or 20 mg / g. As a result, the dermatitis score, epidermal thickness, and blood IgE concentrations increased by butanol fraction (CBBF) due to atopic dermatitis (FIG. 2, FIG. 3).

3. Atopic Dermatitis According to Period Butanol  Fraction CBBF ) Effect

In order to observe the improvement effect of butanol fraction (CBBF) according to the induction period of atopic dermatitis, the induction period of atopic dermatitis was extended to 6 weeks, and the butanol fraction (CBBF) was applied to the affected area for 12 days at a concentration of 10 mg / g. The same result was obtained.

1) Gross and histological changes of the affected area: As shown in FIG. 4, when the induction period of atopic dermatitis was extended to 6 weeks, the dermatitis score did not increase slightly, but the epidermal layer thickness increased significantly. Treatment with butanol fraction reduced both the dermatitis score and epidermal layer thickness, and caused both measurements for 6 weeks (54% decrease in dermatitis score, 64% decrease in epidermal thickness) (42% decrease in dermatitis score, epidermal layer). 58% reduction in thickness).

2) Changes in Serum IgE Concentration and Serum Histamine Concentration: As shown in FIG. 5, both serum IgE and serum histamine concentrations were significantly increased by prolonging the atopic dermatitis induction period for 6 weeks. Treatment with butanol fraction reduced both serum IgE and serum histamine levels, both measurements induced for 6 weeks (69% decrease in IgE, 70% reduction in histamine) (52% decrease in IgE, 40% histamine 40%). Decrease).

1 is a view showing the extraction process of ethanol extract (CBEE) and butanol fraction (CBBF) of the present invention from yellow bunch Cordyceps sinensis,

Figure 2 shows the effect of butanol fraction (CBBF) on each dermatitis index increased by atopic dermatitis. The longitudinal unit of Epidermal Thickness is μm and the serum IgE unit is μg / ml.

NC: normal group, AC: atopy group, CBBF 5, 10, 20 mg / g administration group. Experimental Value: 6 Means ± Standard Error.

Figure 3 is a photograph showing the effect of butanol fraction (CBBF) on the histological changes of the skin caused by atopic dermatitis.

A: normal group, B; Atopy group, C: CBBF 10 mg / g administration group. Magnification × 100, scale bar = 100 μm.

4 is a graph showing the effect of butanol fraction (CBBF) on the increased dermatitis score and epidermal layer thickness by atopic dermatitis.

NC: normal group, AC: atopy group, CBBF 10 mg / g administration group. Experimental Value: 6 Means ± Standard Error.

5 is a graph showing the effect of CBBF on serum IgE and histamine concentrations increased by atopic dermatitis.

NC: normal group, AC: atopy group, CBBF 10 mg / g administration group. Experimental Value: 6 Means ± Standard Error.

Claims (10)

Cordyceps for the prevention and treatment of skin pruritus and atopic dermatitis alcohol extract of bassiana ). The method of claim 1, The alcohol may be one or more alcohols selected from ethanol, propanol and butanol, and it is Cordyceps for preventing and treating skin pruritus and atopic dermatitis. alcohol extract of bassiana ). The method of claim 1, The alcohol extract is a butanol fraction obtained by adding water: butanol mixed solvent to the water fraction obtained by extracting the ethanol extract with a water: hexane mixed solvent, and it is Cordyceps for preventing and treating skin pruritus and atopic dermatitis. alcohol extract of bassiana ). Yellow tuft fungus (Cordyceps A composition for preventing and treating skin pruritus and atopic dermatitis containing an alcoholic extract of bassiana ). The method of claim 4, wherein The alcohol is yellow bundle fungus, characterized in that ethanol, propanol, and the at least one alcohol selected from butanol (Cordyceps A composition for preventing and treating skin pruritus and atopic dermatitis containing an alcoholic extract of bassiana ). The method of claim 4, wherein The alcoholic extract is an ethanol extract of water: hexane fraction obtained by extracting the water was added to a mixed solvent: yellow bundle Cordyceps of butanol fraction of said extract is obtained by adding the butanol mixed solvent (Cordyceps A composition for preventing and treating skin pruritus and atopic dermatitis containing an alcoholic extract of bassiana ). The method according to any one of claims 4 to 6, The composition is characterized in that the composition for external application to the skin, yellow bunch Cordyceps A composition for preventing and treating skin pruritus and atopic dermatitis containing an alcoholic extract of bassiana ). Yellow tuft fungus (Cordyceps Cosmetic composition for improving skin pruritus and atopic dermatitis containing an alcohol extract of bassiana ). The method of claim 8, The alcohol is yellow bundle fungus, characterized in that ethanol, propanol, and the at least one alcohol selected from butanol (Cordyceps Cosmetic composition for improving skin pruritus and atopic dermatitis containing an alcohol extract of bassiana ). The method of claim 8, The alcoholic extract is an ethanol extract of water: hexane fraction obtained by extracting the water was added to a mixed solvent: yellow bundle Cordyceps of butanol fraction of said extract is obtained by adding the butanol mixed solvent (Cordyceps Cosmetic composition for improving skin pruritus and atopic dermatitis containing an alcohol extract of bassiana ).

Images