KR101121514B1 - Anti atopic dermatitis containing extracts from Agrimonia pilosa as active ingredients - Google Patents

Abstract

The present invention relates to a cosmetic composition for anti-atopic dermatitis, and more particularly, to an anti-atopic cosmetic composition containing a straw shinnaum extract and an active ingredient thereof. The cosmetic composition according to the present invention can be effectively used to improve atopic dermatitis by effectively improving atopic symptoms by showing erythema improvement, itching, and sedation effect through cytokine secretion control and immune suppression. .

Anti-atopic dermatitis

Description

Anti atopic dermatitis containing extracts from Agrimonia pilosa as active ingredients

The present invention relates to a cosmetic composition for anti-atopic dermatitis, and more particularly, to an anti-atopic cosmetic composition containing a straw shinnaum extract and an active ingredient thereof.

Atopy dermatitis is an allergic disease that aggravates the stratum corneum, the outermost skin barrier, due to genetic and environmental immunological causes. Atopic dermatitis is a complex entanglement of a variety of causes, repeated relief and recurrence. Allergic diseases caused by atopy predisposition include allergic dermatitis, allergic rhinitis, asthma, allergic conjunctivitis, and atopic urticaria.

The exact pathophysiology of the atopic dermatitis is not yet fully understood, but it is thought that immunological and non-immunological mechanisms may be involved with genetic predisposition. Exogenous atopic dermatitis, which accounts for the majority of atopic dermatitis, is caused by an immune mechanism associated with immunoglobulin E (IgE), and there are many reports that the primary immune response is caused by T-cell abnormalities rather than a secondary immune response to a particular allergen. As cell-mediated immune dysfunction is associated with an increase in immunoglobulin E in patients with atopic dermatitis, there are increasing reports claiming that the primary immune response is involved in the pathogenesis of atopic dermatitis in addition to the secondary immune response (Kee Young, et al. , Korean Medicine, Seoul, 2001, pp23 ~ 52).

It is known that cell-mediated immune disorders occur in patients with atopic dermatitis up to about 80%, and sensitivity to skin infections caused by viruses and skin filaments is increased and sensitivity to contact allergens tends to decrease. In the past, T-cell maturation deficiency and the decrease of CD8 + inhibitory T-cells have been reported, but recently, the role of CD4 + T-cells is known to be more important. In other words, T-cells secreting interleukin 4, 5 that induces or promotes the production of immunoglobulin E from B cells, and interleukin 6 that amplify are Th2 cells among CD4 + T-cells, which are Th1 cells in the skin of atopic patients. It was found that Th2 cells were present higher than that of the cells. These cytokines stimulate immune cells such as mast cells, macrophages and basophils to promote secretion of histamine, leukotrien, prostaglandin and nitric oxide (NO). These inflammatory mediators are known to promote skin inflammation. Another characteristic of atopic dermatitis is the lack of gamma-linolenic acid in lipids that form the cell membranes of skin cells. Gamma linolenic acid deficiency is a phenomenon caused by impaired synthesis of linoleic acid to gamma linolenic acid due to a loss of function of delta-6-desaturase in the biosynthetic pathway of essential fatty acids, followed by prostaglandins Do not control the synthesis of the back, the production of inflammatory mediators will increase. In addition, the deficiency of gamma linolenic acid causes abnormalities in the structure of the skin lipid membrane, resulting in excessive water leakage, resulting in the breakdown of the skin barrier, and at this time, secondary infection such as bacteria occurs.

In view of the etiology, mechanisms, and symptoms of atopic dermatitis as described above, a therapeutic agent for atopic dermatitis has been developed. As a result, a number of natural or synthetic immunosuppressive agents, antihistamines, steroid preparations, and the like have been developed. For example, immunosuppressants include Cyclosporin A, FK506 (Tacrolimus, Tacrolimus; Wasik et al., Immunopharmacology 20: 57-61, 1990) and SDZ ASM981 (Pimecrolimus, Pimecrolimus; Grassberger et al., Br. J. Dermatology 141: 264-273, 1999). On the other hand, steroids and antihistamines, which are used as treatments for atopic dermatitis, have the effect of temporarily relieving the symptoms, but skin thinning and osteoporosis in patients with long-term use of external or oral steroid hormones. In children, it is clinically problematic due to local and systemic side effects such as growth inhibition. Therefore, there is a need to develop a therapeutic agent having no such side effects and excellent efficacy.

The cause of this atopic dermatitis is not well known, but it has been found to be mainly related to genetic factors and immune system deficiency. In addition, dry skin, the characteristics of skin itching easier than normal people, infections caused by bacteria, viruses, fungi, etc., emotional factors, environmental factors seems to be caused by a combination of each other.

The main symptoms of atopic dermatitis are severe itching, dry skin, rashes, rashes, crusts and scaly skin (in scales). Above all, itching is characterized by severe itching.

Because atopic dermatitis cannot be fundamentally repaired, atopic dermatitis is a disease that is controlled by avoiding the triggers and appropriate treatment rather than aiming at cure. Prescriptions for atopic dermatitis are mainly drug therapy such as steroids, antihistamines and antibiotics. Steroids (adrenal corticosteroids) have anti-inflammatory and immunosuppressive effects and have excellent effects, but when applied for a long time, side effects such as skin weakness, systemic hormonal symptoms, and addictive effects may occur. Antihistamines prevent the release of histamine from mast cells and reduce itchy symptoms, but may be used as a temporary measure, such as insomnia, anxiety, and loss of appetite.

Therefore, in order to treat such atopic dermatitis, the antibacterial, moisturizing, and anti-inflammatory effects are excellent, and there is an urgent need for the development of a cosmetic material that can easily use a product containing the material and have no side effects and low cost.

In view of the above, the inventors of the present invention, while conducting research on natural products that can treat atopic dermatitis, observed that straw extract exhibits cytokine secretion regulation and immunosuppression, and thus, atopy-induced animals As a result of applying to the subjects confirmed that the atopic symptoms are significantly alleviated, the present invention was completed by confirming that the straw extract of the present invention can be usefully used in atopic diseases.

An object of the present invention is to provide a cosmetic composition for anti-atopic dermatitis containing Agrimonia pilosa extract as an active ingredient.

The present invention provides a straw herb extract extracted from ethanol in order to achieve the above object.

In addition, the present invention is a straw herb ( Agrimonia pilosa ) provides an anti-atopic cosmetic composition containing an extract as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention is characterized by providing an anti-atopic dermatitis (Agrimonia pilosa) extract for improving atopic dermatitis, wherein the extract is characterized in that obtained from the ethanol extract.

The straw herb extract may be selected from the group consisting of methanol, ethanol, isopropyl alcohol and butanol, preferably methanol or ethanol, and more preferably ethanol.

As a specific example, the straw shinnaum extract is preferably carried out at 60 ℃ to 80 ℃ temperature conditions using the 100% ethanol to crushed straw shinsa, extraction time may vary depending on the temperature conditions during extraction Extract by stirring slowly for 1 day.

The present invention is a straw herb ( Agrimonia pilosa ) provides an anti-atopic cosmetic composition containing an extract as an active ingredient.

The cosmetic composition may be effectively used for atopic improvement or treatment through cytokine secretion regulation and immune suppression. In order to confirm the usefulness of the extract of hay fever, experiments on cytokine secretion inhibitory activity and immune response inhibitory activity were performed.

The pro-inflammatory cytokines are interleukin-4, interleukin-5, interleukin-6, interleukin-8, interleukin-13, interferon gamma (IFN-γ), monocyte chemoattractant protein-1 (MCP- It is characterized in that it is selected from the group consisting of 1).

Specifically, the straw extract of the present invention inhibits the expression of pro-inflammatory cytokines, and is a monocyte chemoattractant compared to the steroid preparation dexamethasone (dexamethasone) as a control of human monocytes (THP-1). It was confirmed that the secretion of the substance protein-1, interleukin-6 and interleukin-8 is effectively suppressed, and also the concentration-dependent inhibition of pro-inflammatory cytokines (Sp. It was confirmed that the inhibitory capacity becomes maximum when it is concentration. This is confirmed that the straw extract according to the present invention has the ability to regulate inflammatory cytokines (pro-inflammatory cytokines).

The straw herb ( Agrimonia pilosa ) extract is characterized in that the suppression of the expression of pro-inflammatory cytokines (regulated) regulates the concentration of immunoglobulin-E (lgE) in the blood.

As a specific example, dinitrochlorobenzene (DNCB) -induced atopic mice were collected from the eye mucosa using capillaries at intervals of about 2 weeks to quantify blood immunoglobulin-E (lgE) in blood. As a result, it was confirmed that the secretion of immunoglobulin-E (lgE) in the blood was more suppressed in the group treated with the dexamethasone (dexamethasone), which was orally administered with the extract of the herbaceous ginseng extract. It was confirmed that it showed excellent immunosuppressive activity. See FIG. 5.

In addition, the improvement of atopic dermatitis was examined by evaluating the degree of dermatitis on the extract of Straw Sprouts using Dinitrochlorobenzene (DNCB) -induced atopic mice. As a result, the degree of dermatitis was remarkably alleviated in the group treated with oral ginseng extract, and the symptom was relieved within a week, mostly in the itching and erythema. In addition to the reduction, sores reduction, etc., the effect persists over time. See FIG. 6.

Straw herb of the present invention ( Agrimonia pilosa ) extract is preferably contained in an amount of 5 to 30% by weight of the total cosmetic composition as an active ingredient, more preferably 10 to 20% by weight.

When using less than 5% by weight of the extract of straw herb contained in the cosmetic composition is difficult to show the effect, when used in excess of 30% by weight may indicate problems such as skin side effects, instability of the formulation.

The present invention can be useful as a therapeutic agent or therapeutic aid for cosmetics for skin diseases such as atopic dermatitis and acne, which is a kind of immune cell mediated inflammatory response caused by an excessive immune response. Dermatitis, allergic dermatitis, eczema, psoriasis, acne and the like can be included.

The cosmetic composition of the present invention is not particularly limited in its formulation, and may be, for example, a cosmetic composition having a formulation of a flexible cosmetic water, a nourishing cosmetic water, a massage cream, a nourishing cream, a pack, a gel or a skin adhesive cosmetic, and Transdermal dosage forms such as lotions, ointments, gels, creams, patches or sprays.

As described above, the extract of straw according to the present invention acts on cytokine secretion control and anti-inhibition, and shows effects such as erythema reduction, itching extinction, antimicrobial activity, deductive inhibition and control effect, and applied to the improvement of atopic dermatitis. This can be usefully used.

The present invention will be described in more detail by the following examples and experimental examples. However, the following Examples and Experimental Examples are only for better understanding of the present invention, and the scope of the present invention is not limited by these Examples and Experimental Examples in any sense.

Hereinafter, the technical and scientific terms used herein will be understood by those skilled in the art without departing from the scope of the present invention. Descriptions of known functions and configurations that may be unnecessarily blurred are omitted.

[ Manufacturing example Straw Sprouts ( Agrimonia pilosa A) Ethanol Extract Manufacturer

After extracting and grinding the dried straw thin herbs, the extraction solvent was mixed and extracted. At this time, a variety of extraction solvents may be used, but 100% ethanol, which is widely used in industry, was selected and used. When the 100% ethanol was mixed with crushed straw vinegar, the mixture was mixed at a ratio of 500 ml of extraction solvent per 30 g of sample, and the temperature condition was performed at 70 ° C., and the extraction time may vary depending on the temperature condition during extraction. The extraction proceeded with stirring. Thereafter, the extraction solution was recovered and concentrated under reduced pressure to completely remove the ethanol solvent. 10 g of the final straw shoots extract obtained in the above process was obtained.

[ Test Example  1] person of straw herb extract In monocytes  Cytotoxicity Check

In order to confirm the cytotoxicity of the extract according to the production of straw shinnamul in human monocytes (THP-1, Bank of the United States Cell Line) treated with untreated, 1 ㎍ / ㎖ straw extract, 10 ㎍ / ㎖ straw extract After 24 hours, cytotoxicity (MTT assay) was confirmed.

The cytotoxicity (MTT assay) is a water-soluble MTT formazan (3- (4,5-dimethylthiazol-2-yl) -2,5 which has a blue purple color of MTT tetrazolium, a yellow water-soluble substrate by dehydrogenase action. This test uses the ability of the mitochondria to reduce to -diphenyl-tetrazolium bromide. The absorbance of MTT formazan is maximal at a wavelength of 540 nm, and the absorbance measured at this wavelength reflects the concentration of live, metabolic cells.

As can be seen in Figure 1, the cytotoxicity of the extract of hay fever did not decrease the cell viability compared to the untreated, which is a result confirming that the toxicity to the cells at the treatment concentration of the present invention.

[ Test Example  2] Inflammatory Cytokines of Extracts from Straw Sprouts pro - inflammatory cytokines Expression suppression confirmation of

(One) Monocytes  Chemical Attractant Protein-1 ( MCP Secretion of -1) Inhibitory ability

RPMI 1640 medium (Life Technologies Inc.) containing 0.5% FBS and antibiotics (100 U / ml penicillin, 100 μg / ml streptomycin) in human monocytes (THP-1). Incubated for 16 hours at 37 ° C.), and treated with dexamethasone (dexamethasone), a steroid preparation, as a control, and treated with 0.1 μg / ml, 1.0 μg / ml, and 10 μg / ml of the straw herb extract prepared in the above preparation. It was.

After 1 hour of treatment with the dexamethasone and hay fever extract, 1 μg / ml of house dust mite extract (DP) was treated for 24 hours, and then monocyte chemoattractant protein-1 in supernatant using ELISA. The amount of (monocyte chemoattractant protein-1, MCP-1) was measured.

As a result, as can be seen in Figure 2, it was confirmed that the secretion of the monocyte chemoattractant protein-1 is effectively inhibited compared to the control, and also the concentration-dependent monocyte chemoattractant protein-1 (MCP-) It was confirmed that the secretion of 1) was suppressed and the inhibitory capacity was maximum when the concentration was 10 µg / ml.

(2) Interleukin -6 secretion Inhibitory ability

RPMI 1640 medium (Life Technologies Inc.) containing 0.5% FBS and antibiotics (100 U / ml penicillin, 100 μg / ml streptomycin) in human monocytes (THP-1). Incubated for 16 hours at 37 ° C.), and treated with dexamethasone (dexamethasone), a steroid preparation, as a control, and treated with 0.1 μg / ml, 1.0 μg / ml, and 10 μg / ml of the straw herb extract prepared in the above preparation. It was.

After 1 hour of treatment with the dexamethasone and straw herb extract, 1 μg / ml of house dust mite extract (DP) was treated for 24 hours, and then the amount of interleukin-6 in the supernatant was measured by ELISA. It was.

As a result, as shown in FIG. 3, it was confirmed that the secretion of interleukin-6 was effectively inhibited compared to the control, and the secretion of interleukin-6 was suppressed in a concentration-dependent manner, and the inhibition was maximized at the concentration of 10 μg / ml. It was confirmed.

(3) Interleukin Secretion for -8 Inhibitory ability

RPMI 1640 medium (Life Technologies Inc.) containing 0.5% FBS and antibiotics (100 U / ml penicillin, 100 μg / ml streptomycin) in human monocytes (THP-1). Incubated for 16 hours at 37 ° C.), and treated with dexamethasone (dexamethasone), a steroid preparation, as a control, and treated with 0.1 μg / ml, 1.0 μg / ml, and 10 μg / ml of the straw herb extract prepared in the above preparation. It was.

After 1 hour of treatment with the dexamethasone and hay fever extract, 1 μg / ml of house dust mite extract (DP) was treated for 24 hours, and then the amount of interleukin-8 in the supernatant was measured by ELISA. It was.

As a result, as can be seen in Figure 4, it was confirmed that the secretion of interleukin-8 is suppressed in a concentration-dependent manner, it was confirmed that the maximum when the inhibitory capacity is 10 ㎍ / ㎖ concentration.

[ Test Example  3] of straw herb extract Anti-allergy  Effect verification

(One) Dinitrochlorobenzene ( DNCB Induction of a) Induced Atopic Mice

After hair removal from 5 to 6 week old Nc / Nga mice, the hair is removed and left for 24 hours to heal fine wounds on the skin. 200 μl of 1% DNCB solution (acetone: olive oil = 3: 1) is applied to the back area. After 4 days, 150 μl of 0.2% DNCB solution was applied to the back site for 4 to 5 weeks, 2-3 times a week to induce anitic mice induced with dinitrochlorobenzene (DNCB).

The induced dinitrochlorobenzene (DNCB) -induced atopic mice were administered orally every day for three weeks at a concentration of 10 mg / kg and 100 mg / kg of dexamethasone, which is a steroid preparation, dexamethasone and 100 mg / kg. After sensory evaluation, the blood is taken and the skin at the back is excised and stored in 10% formalin solution. The blood precipitated the blood cells, and only serum was taken and stored frozen at -70 ° C.

(2) blood immunoglobulin-E ( lgE Secretion for Inhibitory ability

Blood was collected from intraocular mucosa using capillary blood at about 2 weeks from dinitrochlorobenzene (DNCB) -induced dinitrochlorobenzene (DNCB) -induced amplification of blood immunoglobulin-E (lgE) in blood. . Usually, when the disease is induced and the clinical score exceeds 6-7 points, the blood immunoglobulin-E (lgE) concentration becomes 10 µg / ml or more, and the collected blood is measured by diluting about 200 to 500 times.

Blood immunoglobulin-E (lgE) concentrations were performed twice at the end of dinitrochlorobenzene (DNCB) treatment using the ELISA method and at the end of treatment of the therapeutic drug. The serum immunoglobulin-E (lgE) concentration in serum was calculated by comparing the blood immunoglobulin-E (lgE) concentration before the drug treatment with the blood immunoglobulin-E (lgE) concentration after the treatment, and then treated with the drug. The significance was verified by comparing with the decrease ratio of.

As can be seen in Figure 5, it was confirmed that the secretion of blood immunoglobulin-E (lgE) in the group treated with the steroid preparation dexamethasone (dexamethasone) in the group orally administered straw shinnamul 10 mg / kg, Considering the fact that it is a natural extract, it can be seen that it shows a very good immunosuppressive activity.

(3) Effect of Straw Sprout Extract on Dermatitis

In order to evaluate the degree of dermatitis in atopic dermatitis induced by dinitrochlorobenzene (DNCB) induced in (1), a clinical visual evaluation method generally used in atopic dermatitis was used. The visual evaluation result is the sum of the scores of each of the following five items. The assessment items are divided into erythema, itching and dry skin, edema and escoriation, erosion and lichenification. Each of these items is scored as none (0), weak (1), severity (2), and severe (3). Therefore, you can get a score between at least 0 and at most 15. In general, when dermatitis is induced using DNCB, it can be determined that the dermatitis peaked at about 12 to 13 points.

Sensory evaluation was performed twice at the end of dinitrochlorobenzene (DNCB) treatment and at the end of treatment of the treatment drug. Scoring of dermatitis by sensory evaluation was performed by two or more experienced subjects, and was determined by consultation.

As a result of evaluating the degree of dermatitis, the treatment was evaluated by comparing the pretreatment score and the posttreatment score.

As can be seen in Figure 6, in the group treated with oral ginseng 10 mg / kg dexamethasone (dexamethasone) in the group treated than the degree of dermatitis was significantly relieved, most of the symptoms within a week to confirm that In addition to reducing itching and erythema and reducing sores, it was confirmed that the effect persists over time.

(4) interferon gamma ( IFN secretion for Inhibitory ability

The spleen cells of the atopy mice induced with dinitrochlorobenzene (DNCB) induced in (1) were isolated, and the amount of interferon gamma (IFN-γ) according to the extract of Streptomyces was measured in the supernatant using ELISA. .

As a result, as can be seen in FIG. 7, the secretion of interferon gamma (IFN-γ) is more suppressed in the group treated with dexamethasone (dexamethasone), which was orally administered with 10 mg / kg of straw shoots. there was.

1 is a result of confirming the cytotoxicity in human monocytes of the extract of straw according to the present invention,

Figure 2 is a result confirming the secretion inhibitory ability of the mononuclear cell chemoattractant protein-1 (MCP-1) in human monocytes according to the treatment concentration of the extract of the extract according to the present invention,

3 is a result confirming the secretion inhibitory ability of interleukin-6 in human monocytes in accordance with the treatment concentration of the extract of the present invention.

4 is a result confirming the secretion inhibitory ability against interleukin-8 in human monocytes in accordance with the treatment concentration of the extract of the straw Shinmul of the present invention,

5 is a result of confirming the secretion inhibitory ability of immunoglobulin-E (lgE) in the blood of adenosine mice induced with dinitrochlorobenzene (DNCB) of the extract according to the present invention,

6 is a result of visual evaluation of the effect on dermatitis in atopic dermatitis induced by dinitrochlorobenzene (DNCB) of the extract according to the present invention,

(●: PBS, ■: Dexa, ▲: Straw Sprout Extract 10mg / kg, ×: Straw Sprout Extract 100mg / kg)

7 is a result of confirming the secretion inhibitory ability of interferon gamma (IFN-γ) in atopic dermatitis-induced dinitrochlorobenzene (DNCB) of the extract according to the present invention.

Claims (6)

A cosmetic composition containing Agrimonia pilosa extract as an active ingredient. The method of claim 1, The Agrionia pilosa extract is a cosmetic composition, characterized in that obtained from the ethanol extract. 3. The method of claim 2, The extract according to Agrimonia pilosa is a cosmetic composition, characterized in that it contains 5 to 30% by weight of the total composition as an active ingredient. The method of claim 3, The extract of Agrionia pilosa is a cosmetic composition, characterized in that to suppress the expression of inflammatory cytokines (pro-inflammatory cytokines). The method of claim 4, wherein The extract of Agrimonia pilosa is a cosmetic composition, characterized in that the suppression of the expression of inflammatory cytokines (pro-inflammatory cytokines) regulates the concentration of immunoglobulin-E (lgE) in the blood. The method according to claim 4 or 5, The pro-inflammatory cytokines are interleukin-4, interleukin-5, interleukin-6, interleukin-8, interleukin-13, interferon gamma (IFN-γ), monocyte chemoattractant protein-1 (MCP- Cosmetic composition, characterized in that it is selected from the group consisting of 1).

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