KR101330686B1 - Composition for prevention or treatment of dermatitis Comprising an extract of herbal combination - Google Patents

Abstract

The present invention relates to a composition for the prevention or treatment of dermatitis diseases, including a mixed extract of bokbunja, eoseongcho, saengjihwang and hides as an active ingredient. Complex extracts of bokbunja, eochocho, raw sulfur and hides of the present invention reduce inflammatory cell invasion of skin tissues, blood IgE (Imnunoglobulin) concentration and IL-4 (InterLeukin-4), IL-13 and IL-17 cytokine secretion It can be usefully used for the prevention or treatment of dermatitis diseases by reducing the.

Description

Composition for prevention or treatment of dermatitis Comprising an extract of herbal combination}

The present invention relates to a composition for the prevention or treatment of dermatitis diseases comprising a composite extract of bokbunja, eoseongcho, saengjihwang and Hwapi as an active ingredient.

The skin of the human body is an indispensable organ for the maintenance of life, which physically and chemically protects the body from extraterrestrial and performs the biochemical functions necessary for metabolism of the whole body. All abnormalities that appear on human skin are called skin diseases, that is, skin diseases. Since the skin covers the surface of the body, there are many opportunities for direct contact with stimuli or various pathogens from the outside, and are strongly influenced by the body. Furthermore, even minor changes in the skin can be seen by eye and touched by hand, and it is easy to examine pathological histologically by extracting a part of the lesion, to examine microorganisms, and to clearly diagnose the diagnosis of individual diseases. However, their types are considerably larger than those of other organs, and their disease names are complicated and diverse. Relatively common skin diseases include atopic dermatitis, contact dermatitis, seborrheic dermatitis, urticaria, athlete's foot, psoriasis, psoriasis, herpes simplex, shingles, dry skin, housewife eczema, acne.

Contact dermatitis is a dermatitis caused by contact with foreign substances and is a kind of eczema. Contact dermatitis has irritant contact dermatitis and allergic contact dermatitis depending on the mechanism in which it occurs, and one substance can cause these two reactions simultaneously. Most of the substances that cause contact dermatitis are organic compounds. Once the skin comes into contact with these substances again, it is thought that memory cells detect them and secrete several chemicals that cause inflammation. Contact dermatitis is often limited to contact areas, so site identification is very important in identifying substances.

Atopic dermatitis is a chronic relapsing dermatitis that occurs mostly in infancy and childhood and improves and worsens. Diagnosis criteria include pruritus, characteristic form and distribution, chronic dermatitis, history of atopic disease, and family history. It is becoming. The prevalence is steadily increasing in Korea, with prevalence of 10-20% in children and 2-4% in adults, increased air exposure, exposure to antigens due to changes in residential conditions, reduced breastfeeding, and childhood infectious diseases. It is believed that the decline, the western way of life, and mental stress are the causes of the increase.

The clinical symptoms of atopic dermatitis may vary from person to person, but the most common symptoms are first, severe itching, second, the patient's skin remains dry, and third, dry skin. Due to the loss of the defense function of the skin surface, it is easy to invade the skin of irritants from the outside, and the invasion causes the skin to reject the inflammatory reaction (Lancet. 1998; 351: 1715-1721, J. Pediatr). Health Care. 2002; 16; 143-145.

The exact pathophysiology of atopic dermatological diseases is not fully understood yet, but it is known that external factors cause overexpression of inflammatory and immune responses in the body. When atopic dermatitis is induced, inflammation-related cells such as eosinophils, neutrophils, and mast cells are observed at the lesion site. Along with this, immunoglobulin antibody E (IgE, Immunoglobulin E) produced from mast cells rapidly increases. In addition, aberrant proliferation of T cells is observed, and in the process, activation of type 2 T-cells and the increase of cytokines, IL-4, IL-5, and IL-13 amplify the immune response. Let's do it. Based on this, research to develop atopic dermatitis therapeutics through inflammation and immunosuppression is increasing.

Many studies have been conducted on the cause and treatment of atopic dermatitis until recently, but due to the complexity of the disease itself and conflicting research data, no clear theory has yet been established on the exact etiology and effective treatment. Currently developed atopic dermatitis treatments include topical anti-inflammatory drugs such as steroids, tacrolimus, pimecrolimus, antihistamines and immunosuppressants such as cyclosporin. Photo chemotherapy is also used in the clinic. However, these therapies or therapies help to control the symptoms to an appropriate level, rather than the fundamental treatment, to date do not fully meet the needs of patients with atopic dermatitis.

Accordingly, many attempts have been made to find therapeutic agents having high therapeutic effects and low side effects from natural products having proven safety. Anti-atopy-related substances known to date include Dansam, Darae, Chamsori, Sambaekcho, Jaungo, and Hwangyeonhaedoktang.

On the other hand, the bokbunja (Rubus Coreanus Miquel) refers to the immature fruit of the bokbunja berries and related plants of the same species of deciduous shrub belonging to the rosaceae, also called a stigma. Rehmannia glutinosa Libschitz var.purpurea Makino refers to the fresh roots of a perennial vegetation belonging to the Hyunsam family. Houttuynia cordata thunb is obtained by drying the ground part of flowering plants of perennial herbaceous herbaceous plants of over three hundred. The beetula cortex is the dried bark of the birch, a deciduous tree belonging to the birch family.

The present inventors continue to search for a therapeutic agent for a dermatitis disease having a high therapeutic effect and low side effects, and have excellent dermatitis when a complex extract of bokbunja, raw yellow sulfur, fish herb and skin is treated by skin application or oral administration in an animal model of skin disease. The present invention has been completed by confirming that the disease exhibits therapeutic efficacy.

It is an object of the present invention to provide a pharmaceutical composition for the prevention or treatment of dermatitis diseases, which is effective in treating dermatitis diseases while having fewer side effects in vivo.

In addition, an object of the present invention is to provide a health functional food for the prevention or improvement of dermatitis diseases having a less effective side effect of dermatitis disease in vivo.

In addition, an object of the present invention is to provide a cosmetic composition for the prevention or alleviation of dermatitis diseases having a less effective side effect of dermatitis disease in vivo.

In order to achieve the object of the present invention as described above, the present invention provides a pharmaceutical composition for the prevention or treatment of dermatitis diseases comprising a composite extract of bokbunja, raw jihwang, eoseongcho and Hwapi as an active ingredient.

In another aspect, the present invention provides a health functional food for the prevention or improvement of dermatitis diseases comprising a complex extract of bokbunja, raw jihwang, eoseongcho and Hwapi as an active ingredient.

In another aspect, the present invention provides a cosmetic composition for the prevention or alleviation of dermatitis diseases comprising a complex extract of bokbunja, raw jihwang, eoseongcho and Hwapi as an active ingredient.

A pharmaceutical composition for the prevention or treatment of dermatitis diseases comprising a complex extract of bokbunja, raw egg yolk, fish vinegar and hides as an active ingredient inhibits inflammation-related cell numbers and suppresses T cell activity and IgE in animal models of contact dermatitis and atopic dermatitis. Inhibition has a marked therapeutic effect on dermatitis diseases. Therefore, the composition may be used in medicines for the prevention or treatment of dermatitis diseases including contact dermatitis and atopic dermatitis or in health functional foods and cosmetics for the prevention or improvement of dermatitis diseases including contact dermatitis and atopic dermatitis. have.

Figure 1 is a graph confirming the change in the weight of the mouse during the acute dermatitis mouse model experiment using DCNB.
2 is a blood analysis result (red blood cells, platelets, leukocytes, lymphocytes, monocytes, neutrophils, eosinophils in acute dermatitis mouse model using DCNB).
Figure 3 is a picture comparing the degree of infiltration of inflammatory cells through hematoxylin & eosin staining of the skin tissue in the acute dermatitis mouse model using DCNB (top) and a picture of the cell number of the skin (bottom).
Figure 4 is a picture analyzing the amount of T cells of the skin through CD3 staining in acute dermatitis mouse model using DCNB.
FIG. 5 is a diagram illustrating the concentration of immunoglobulin E (IgE) in blood in acute dermatitis mouse model using DCNB.
6 is a diagram illustrating the expression level of cytokines (IL-4, IL-6, TNF-α, IFN-γ) in acute dermatitis mouse model using DCNB.
Figure 7 is a diagram comparing the degree of infiltration of inflammatory cells through hematoxylin & eosin staining of skin tissue in a chronic dermatitis mouse model using OVA.
FIG. 8 is a diagram comparing the expression levels of mast cells through toluidine blue staining in skin tissues in a chronic dermatitis mouse model using OVA (top) and quantifying them (bottom).
Figure 9 is the result of blood analysis (eosinophils, neutrophils, lymphocytes, monocytes, total leukocyte order) in the chronic dermatitis mouse model using OVA.
FIG. 10 is a diagram illustrating the concentration of immunoglobulin E (IgE) in blood in a chronic dermatitis mouse model using OVA.
11 is a diagram analyzing the expression level of cytokines (IL-4, IL-13, IL-17) in a chronic dermatitis mouse model using OVA.

Hereinafter, the present invention will be described in more detail.

The present invention provides a pharmaceutical composition for the prevention or treatment of dermatitis diseases comprising a composite extract of bokbunja, raw jihwang, eoseongcho and Hwapi as an active ingredient.

Composite extracts of bokbunja, raw jihwang, eochocho and hupi used in the present invention can be obtained as follows.

Bokbunja, raw sulfur, fish vinegar and hides are washed with water to remove foreign substances, dried in the shade and crushed. Bokbunja, raw sulfur, fish vinegar and bark may be used without limitation, such as those grown or marketed. Appropriate amount of solvent is added to the ground bokbunja, raw sulfur, fish vinegar and hides to ensure complete immersion. The extraction solvent is preferably a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, more preferably ethanol. Next, the extracts of Bokbunja, Saenghwanghwang, Eochocho and Hwapi obtained above were filtered and concentrated under reduced pressure to obtain a final extract.

In the composition according to the present invention, the mixing ratio of the bokbunja, raw sulfur, fish vinegar and hides of the complex extract is 10% to 50%: 10% to 50%: 10% to 50%: 10% to 50%: 10 % To 50% by weight, preferably 15 to 30%: 15 to 30%: 15 to 30%: 15 to 30% by weight. If the mixing ratio is out of the range, there is no significant difference in allergic reactions or inflammatory reactions compared to the case of using the bokbunja, raw jihwang, eoseongcho, Hwapi, Angelica extract alone, there is a concern of side effects In the above composition ratio, the inhibitory effect of inflammatory cytokines is best.

Dermatitis diseases defined in the present invention include atopic dermatitis, contact dermatitis, seborrheic dermatitis, urticaria, athlete's foot, gland, psoriasis, herpes simplex, shingles, dry skin, housewife eczema, acne, and more preferably contact Dermatitis or atopic dermatitis.

Pharmaceutical composition for the prevention or treatment of dermatitis diseases comprising a composite extract of bokbunja, raw jihwang, eochocho and chapi of the present invention as an active ingredient, 0.1 to 90% by weight of the composite extract, more preferably based on the total weight of the composition 0.1 to 50% by weight. However, the composition is not limited thereto, and may vary depending on the condition of the patient, the type of disease, and the progress of the disease.

The composition of the present invention can be used with a complex extract of bokbunja, saengjihwang, Echochocho and Hwapi as an active ingredient, as well as the previously known preventive or therapeutic agent for skin diseases.

The pharmaceutical composition comprising a composite extract of bokbunja, raw jihwang, eoseongcho and Hwapi as an active ingredient of the present invention may be prepared using a pharmaceutically acceptable and physiologically acceptable adjuvant in addition to the active ingredient. Excipients, disintegrants, sweeteners, binders, coatings, swelling agents, lubricants, lubricants or flavoring agents and the like can be used. In addition, the pharmaceutical composition may be preferably formulated into a pharmaceutical composition including one or more pharmaceutically acceptable carriers in addition to the active ingredient described above for administration.

The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include but are not limited to natural and synthetic gums such as starch, gelatin, glucose or beta-lactose, corn sweeteners, acacia, trackercance or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like. Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile water and sterile water suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

An effective amount of the composition refers to an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, doctor or other clinician, which is the disease or disorder being treated. Includes amounts that induce relief of symptoms. It will be apparent to those skilled in the art that the therapeutically effective dose and the number of administrations of the active ingredient of the present invention will vary depending on the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like. In the treatment method of the present invention, in the case of an adult, when administering the complex extract of Bokbunja, Saenghwang, Echocho and Hwapi of the present invention once or several times a day, it is administered at a dose of 1 mg / kg to 250 mg / kg desirable.

The composition comprising the complex extracts of the bokbunja, saenghwanghwang, eoseongcho and chapi of the present invention as an active ingredient, oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes Can be administered in a conventional manner.

In another aspect, the present invention provides a health functional food for the prevention or improvement of dermatitis diseases comprising a complex extract of bokbunja, raw jihwang, eoseongcho and Hwapi as an active ingredient.

The food composition according to the present invention can be formulated in the same manner as the above pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gums, ice creams, alcoholic beverages, vitamin complexes, health supplements, and the like. .

The amount of the complex extract of Bokbunja, raw sulfur, fish vinegar and bark in food or beverage can be added at 0.01 to 15% by weight of the total food weight, and the health functional beverage composition is 0.02 to 10g, preferably 0.3 to 1g based on 100 ml. It can be added at the ratio of.

The food composition of the present invention may contain complex extracts of bokbunja, raw sulfur, fish vinegar, and hides, which are active ingredients, and may contain various flavors or natural carbohydrates as additional ingredients, as in general food compositions. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. The aforementioned flavoring agents can advantageously be used natural flavoring agents (tautin), stevia extracts (for example rebaudioside A, glycyrzin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.).

In addition, the food composition is not only complex extracts of bokbunja, raw sulfur, fish vinegar and hides, but also various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, such as flavoring agents, coloring and neutralizing agents (cheese, chocolate, etc.), Pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks and the like. In addition, the food composition of the present invention may contain natural fruit juice and fruit flesh for the production of fruit juice beverages and vegetable beverages.

Complex extracts of bokbunja, raw jihwang, eoseongcho and Hwapi as the active ingredient of the present invention is a natural substance with little toxicity and side effects, so it can be used with confidence even for long-term administration for skin disease prevention purposes.

In another aspect, the present invention provides a cosmetic composition for the prevention or improvement of dermatitis diseases comprising a composite extract of bokbunja, raw jihwang, eoseongcho and Hwapi as an active ingredient.

 The cosmetic composition according to the present invention may be formulated as an external preparation for skin or cosmetics. The external preparation for the skin may include an ointment, a warning agent, a spray, a suspension, an emulsion, a cream, a gel, and the like.

In addition, the cosmetic composition of the present invention, in addition to the above-mentioned active ingredient, various components generally used in the external preparation for skin within the range of not lowering the effect of the present invention, if necessary, for example, water-soluble components, powder components, oil, surfactants, It is possible to mix | blend a moisturizer, a viscosity modifier, a preservative, antioxidant, a fragrance | flavor, and a pigment | dye. Also, vitamin A derivatives such as moisturizers such as ceramides or lipids, steroids such as hydrocortisone, retinyl palmitate or retinyl palmitate, which have been widely used in conventional atopic skin treatments. And tocopherol (Tocopherol), and other plant extracts can be used in combination with N-stearoyl-phytosphingosine in the form of liposomes (liposome) using a known lecithin or (Nstearoyl-phytosphingosine).

Formulation of the cosmetic composition of the present invention can be selected arbitrarily, conventional cosmetic formulations such as skin ointment, essence, whitening cream, lotion, emulsion, pack, general cosmetics, skin milk, cream, serum, beauty soap, softening cosmetics, medicinal It can be prepared in various forms, such as lotion, hair tonic, systemic cleanser and oil gel. Cosmetic composition of the present invention further comprises at least one additive selected from the group consisting of oil, water, surfactants, moisturizers, thickeners, chelating agents, pigments, preservatives, and fragrances to produce a cosmetic composition, characterized in that Can be.

Hereinafter, preferred preparation examples and examples and experimental examples are presented to help understand the present invention. However, the following Preparation Examples and Examples and Experimental Examples are provided only to more easily understand the present invention, and the contents of the present invention are not limited by the Preparation Examples, Examples and Experimental Examples.

Manufacturing example  1. Preparation of complex extract

Echoweed used in this experiment ( Houttuynia cordata Thunberg ), Bokbunja ( Rubus coreanus ), Raw fish ( Rehmannia glutinosa , Prunus yedoensis ) was purchased from OmniHub Corporation (Korea). Each medicine was pulverized into fine powder using a blender, and then 100 g of the powdered sample was quantified and sonicated with 1 L of 80% ethanol for 30 minutes using an Ultrasonicator. The extracted solution was filtered using filter paper (Whatman, UK), and the remaining medicine was sonicated for 30 minutes by adding 1 L of 80% ethanol again. The extract was concentrated at 45 ° C. using a vacuum concentrator, and the concentrate was lyophilized. After lyophilization, the medicine is blocked and finely ground using a bowl to make powder. Each medicinal herb in powder state was added to saline and dissolved using a homogenizer, and then used for experiments. Powdered medicines were stored frozen and used whenever needed. The amount of each medicine was made by mixing the same amount of bokbunja, raw yellow sulfur, eoseongcho, Hwapi each experimental drug.

Example  One. DNCB (2,4- dinitrochloro benzene Model design for acute dermatitis

1.1 Preparation of Experimental Animals

The experimental animals received 12 male Balb / c mice aged 6 weeks from Orient (Korea). A weekly temperature (24 ± 2 ° C), humidity (40 to 60%), and day and night cycles (12 hours each) were used to adjust the environment by ingesting solid feed and water at the Kyung Hee University College of Oriental Medicine. Three cages were allocated to each cage and divided into four cages.

1.2 DNCB (2,4- dinitrochloro benzene ) To induce skin diseases

Three balb / c mice adapted for 1 week in a laboratory environment were divided into four groups, a normal group, a control group, a skin coating group (DAHM), and an oral administration group (OAHM). After depilation, DNCB was treated to cause acute dermatitis. However, the normal group applied acetone, which is a solvent used to dissolve the DNCB, in place of the DNCB. DNCB (Dinitrochlorobenzene) is a representative substance that causes delayed contact dermatitis. It is known that repeated exposure of DNCB causes skin swelling and thickening of skin tissue due to infiltration of CD4 + and CD8 + T cell mediated inflammatory cells. Sexual dermatitis animal models are currently one of the most widely used animal models (Nakagawa et al., J. Invest. Dermatol., 57, pp 369-377, 1971; Oka et al., Dermatologica., 172, pp 12-17, 1986; Friedmann, Adv. Dermatol., 5, pp175-195, 1990; Upadhye and Maibach, Contact Dermatitis, 27, pp281-286, 1992; Bronaugh et al., Food Chem.Toxicol., 32, pp113-117, 1994 Kolde, Exp.Dermatol., 3, pp 269-275, 1994). It is known that thickening of skin tissue with histologically significant inflammatory cell infiltration is observed (J et al., Vet. Pathol., 36, pp 42-50, 1999). In addition, skin lesions that occur after DNCB application are strictly referred to as atopic dermatitis-like contact hypersensitivity, or atopic dermatitis-induced immune response such as an increase in IgE and Th2 cytokines. have.

DNCB sensitization was performed twice a week. In the first week, 2% of DNCB was used, and after a week of rest, 0.2% DNCB was treated twice a week, causing acute dermatitis. Then, after a week of rest period, DNCB was treated twice a week while prescribing medication to maintain the symptoms of dermatitis.

Example  2. Ovalbumin  ( ovalbumin , OVA Design of animal model for chronic dermatitis

Seven-week-old female Balb / c mice maintained in the same environment as in Example 1.1 were intraperitoneally mixed with 20 μg of ovalbumin (OVA) and 4 mg of Alu (Aluminum hydroxide hydrate) in saline for three weeks once a week. Administration to induce atopic dermatitis sensitization. OVA skin sensitization model takes a relatively long time to induce compared to chemical skin irritation model, and the actual symptoms of dermatitis are difficult to see visually. After 3 weeks of OVA intraperitoneal administration, 100 μg of OVA and 20 mg of Alu were mixed to the skin to induce dermatitis on the skin by sensitizing three times per week for 3 weeks with a 1 × 1 cm patch. After the induction of atopic dermatitis, the composite extracts HM-A (fish vinegar, bokbunja, raw sulfur, skin) and HM-B (including donkey instead of bark) prepared in the manner of Preparation Example 1 were applied for 2 weeks for 2 weeks after patch sensitization. Oral administration at a concentration of 50 mg / kg once. Animal group was normal group (N = 4), which did not induce atopic dermatitis, control group (OVA group; N = 4) that induced atopic dermatitis, and experimental group (HM-A group, HM- orally administered with multiple extracts). The experiment was divided into four groups with B group; each N = 4).

Experimental Example  One. DNCB (2,4- dinitrochloro benzene Therapeutic Effects in Animal Models of Acute Dermatitis

1.1. Weight change

Mouse weight changes were measured during the experimental period of Example 1.

As shown in FIG. 1, no significant weight change was seen by the drug. Therefore, it was confirmed that the therapeutic efficacy of skin diseases was not due to the change in weight afterwards.

1.2 Blood Analysis

After the two-week experimental period of Example 1, blood collected from the heart of the mouse was analyzed using a blood analyzer to observe changes in erythrocytes, platelets, leukocytes, lymphocytes, mononuclear leukocytes, neutrophils, and eosinophils in the mice.

Changes in blood components are indicative of whether or not dermatitis, in particular, atopic dermatitis is invented. Hematological changes in atopic dermatitis are mostly increased by immune-related cells due to an inflammatory response. Among them, platelets are cells involved in innate immunity in the immune response and are activated by allergens to induce an inflammatory response. In addition, white blood cells have a role of decomposing bacteria or foreign substances invaded from the outside, the number of white blood cells increases when inflammation is expressed. In addition, after a certain period of time after the production of immune cells, they should be destroyed by apoptosis naturally. In patients with dermatitis diseases such as atopic dermatitis, cell death of immune cells is inhibited by cytokines and the like, and lymphocytes increase, resulting in diseases such as atopic dermatitis. . Mononuclear leukocytes (monocytes) also play an important role in both intrinsic and adaptive immunity and are responsible for differentiating macrophages and antigen presenting cells. However, excessive mononuclear leukocytes can cause severe inflammation. Neutrophils are the most granular white blood cells in the blood and play an important role in antibacterial and antibacterial activity, but they also mediate tissue destruction in inflammatory diseases such as rheumatoid arthritis. Therefore, excessive amounts of neutrophils can cause inflammatory reactions, which can lead to more severe dermatitis symptoms. Finally, eosinophils are granular leukocytes that mainly act on immunity such as parasites and are generally associated with atopic diseases and mast cells. Eosinophils contain major basic protein (MBP), which is highly toxic to tissues and is markedly observed in atopic dermatitis.

As shown in Figure 2, there was no change in the amount of red blood cells in all groups, it was confirmed that the level of platelets in the drug administration group. In addition, the levels of leukocytes, lymphocytes, monocytes, neutrophils, and eosinophils increased by DNCB were significantly decreased in the normal range in the drug treatment group (skin application group (DAHM), oral administration group (OAHM)).

1.3 Histological Analysis

Mouse skin tissue of Example 1 was analyzed via hematoxylin & eosin (H & E) staining and CD3 staining. In general, it is known that the amount of T cells is increased in inflammatory diseases such as atopic dermatitis.

As shown in FIG. 3, as a result of H & E staining, it was confirmed that the epidermis and dermis were thickened in the model in which dermatitis disease was invented by DNCB, compared to the normal group, and the number of cells stained with hematoxylin in the skin layer was normal. It was measured about four times higher than the group. On the contrary, it was confirmed that the dermatitis symptom was alleviated because the number of cells of about 3 times in the skin application group and about 2 times in the oral administration group was confirmed.

In addition, as shown in Figure 4, the amount of T cells stained with CD3 can be confirmed that the skin coating group (DAHM), oral administration group (OAHM) of the drug is reduced compared to the control group, the drug is excessive T cells It was confirmed that the inhibition of the production of.

1.4. In the blood IgE  analysis

IgE (immunoglobulin E) is an immunoglobulin that appears in high concentrations in allergic or parasitic infection patients and is known to be present in high concentrations in patients with dermatitis diseases such as atopic dermatitis. In addition, Eosinophil, basophil, and mast cells are combined to sensitize the cells, and allergens are combined to degranulate and secrete bioactive substances such as histamine.

As a result of confirming the amount of IgE in the blood of the mouse of Example 1, as shown in Fig. 5, the amount of IgE in the blood was increased in the control group compared to the normal group, but the drug group (DAHM), oral administration group (OAHM) ), All of them decreased, especially in the oral group.

1.5. Analysis of Cytokine Expression in the Spleen

MRNA was extracted from spleen cells of the mouse of Example 1, IL-4 (interleukin-4) and TNF (tumor necrosis factor) -α, which are the major cytokines for Th 2 immune response, and IL- which are highly related to inflammatory response. MRNA expression levels of IFN (interferon) -γ, a cytokine of the 6 and Th 1 immune responses, were compared. Th1 and Th2 cells interact to maintain immune balance, with IL-4 inhibiting the Th1-type response and IFN-γ inhibiting the Th2-type response. T cells of patients with dermatitis diseases, such as atopic dermatitis, have a decreased ability to produce IFN-γ and have a strong reactivity to IL-4. In atopic dermatitis, an immune response by Th2 type cells is considered to be the main factor. In addition, various cytokines such as IL-5 and IL-13 are involved in causing atopic dermatitis, which promotes epithelial hyperplasia and the expression of granulocytes macrophage colony stimulating factor (GM-CSF). IL-4 plays a major role in the activation of Th2 cells and promotes the production of IgE. IL-6 is a proinflammatory cytokine secreted by macrophages. It is rapidly expressed in the presence of wounds or infections, and when excessively expressed, promotes and sustains the inflammatory response, leading to inflammatory diseases. TNF-α affects atopic dermatitis by acting on the production of various cytokines, cell growth and differentiation.

As shown in FIG. 6, IL-4, IL-6, and IFN-γ did not show significant changes in the skin application group (DAHM) and oral administration group (OAHM) of the drug compared to the control group, but TNF-α. It was confirmed that the expression was reduced compared to the control.

Experimental Example  2. Ovalbumin  ( ovalbumin , OVA Treatment effect in chronic dermatitis induced animal model

2.1. Histopathological evaluation

Skin tissue extracted from the mouse of Example 2 was washed with saline, and then OCT frozen tissue sections were made to hematoxylin & Eosin (H & E) staining and toluidine blue staining mast cells. Staining was done. Mast cells are inflammatory cells involved in immediate allergic reactions in inflammation-related diseases such as asthma and atopic dermatitis, and are key inflammatory cells leading the chain reaction of allergic reactions. Fc receptors that can bind IgE to the mucous membranes of mast cells It is characterized by including histamine and the like. The stained tissues were observed for inflammatory cell infiltration and sensitization of mast cells under 400 magnification. The degree of infiltration of mast cells and inflammatory cells was expressed as cells / HPFs by counting 5 field / N high power fields (HPFs) under 1000 magnification.

As shown in FIG. 7, as a result of performing H & E staining to confirm the invasion of inflammatory cells as a whole, the invasion of inflammatory cells was significantly increased in the OVA group compared to the control group, whereas the experimental group HM-A (eosancho, bokbunja) , Saengjihwang, hides) was confirmed to decrease compared to the OVA group.

In addition, as shown in Figure 8, mast cells were significantly increased in the OVA group compared to the control group, and the expression of mast cells in HM-A (eojecho, bokbunja, fresh yellow, skin) was suppressed.

Through this, it was confirmed that the administration of the drug reduces the infiltration rate of inflammatory cells in the skin tissue and reduces the mast cells, which are the core cells of the allergic reaction, thereby reducing the inflammatory response of the animal's actual skin tissue.

2.2. Blood analysis

Mouse blood of Example 2 was collected and tested for total white blood cell (WBC) levels using an automated hematology analyzer HEMAVET 950 (Drew Scientific, UK).

As shown in FIG. 9, the levels of eosinophils expressed in the blood of atopic dermatitis patients were more than doubled in the OVA group compared to the control group, and HM-A (eosancho, bokbunja, saenghwanghwa, Hwapi) and HM -B (eosancho, bokbunja, saengjihwang, Angelica) decreased compared to the OVA group. In particular, the HM-A (eosancho, bokbunja, raw sulfur, namulhwa) group showed a similar level of reduction as the control. In addition, neutrophils, lymphocytes, and monocytes, which are increased during the inflammatory response, also decreased in HM-A. However, the total WBC was increased in the OVA group compared to the control group, but there was no decrease in drug administration.

2.3. In the blood IgE  analysis

As a result of analyzing the amount of IgE from the blood of the mouse of Example 2, as shown in FIG. It was confirmed that the decrease.

2.4 Cytokine Expression Analysis

Expression of type 2 cytokines IL-4, IL-13 and IL-17 associated with increasing type 2 T cells in atopic dermatitis patients was analyzed. The results were corrected by GAPDH, a housekeeping gene. Representative cytokines associated with type 1 T cells include IFN-γ, and cytokines associated with type 2 T cells include IL-4, IL-5, IL-6, IL-10, IL-13. In addition, IL-17 used in this experiment is a cytokine associated with CD4 helper T cells, and CD4 is a glycoprotein, a helper T cell, a macrophage, a monocyte, and a dendritic cell. It is a protein that is expressed on the surface of is also a cytokine known to increase expression in the immune process.

As shown in FIG. 11, the cytokine was increased in the OVA group compared to the control group, and the results were decreased in both HM-A (eosancho, bokbunja, saenghwanghwang, Hwapi) and HM-B (eosancho, bokbunja, saenghwanghwang, Angelica) It was. In particular, IL-13 showed a lower expression level in HM-A (eosancho, bokbunja, fresh sulfur, namul) than the control.

Hereinafter, the preparation of pharmaceutical compositions, health functional foods, and cosmetic compositions containing the composition of the present invention will be described, but the present invention is not intended to limit the present invention.

Formulation example  1: Preparation of pharmaceutical preparation

One. Sanje  Produce

2g complex extract of Bokbunja, Eoseongcho, Saengjihwang and Hwapi

1g lactose

The above components were mixed and packed in airtight bags to prepare powders.

2. Preparation of tablets

100mg Complex Extract of Bokbunja, Eoseongcho, Saengjihwang and Hwapi

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

3. Preparation of Capsule

100mg Complex Extract of Bokbunja, Eoseongcho, Saengjihwang and Hwapi

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

Formulation example  2 ; Manufacture of food

1. Preparation of dietary supplements

100 mg of complex extract of Bokbunja, Eoseongcho, Saengjihwang and Hwapi

Vitamin mixture quantity

Vitamin A acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

Vitamin B12 0.2 μg

Vitamin C 10 mg

Biotin 10 μg

Nicotinic acid amide 1.7 mg

Folic acid 50 μg

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

2. Health drink  Produce

100 mg of complex extract of Bokbunja, Eoseongcho, Saengjihwang and Hwapi

Vitamin C 15 g

Vitamin E (powder) 100 g

19.75 g of ferrous lactate

3.5 g of zinc oxide

Nicotinic acid amide 3.5 g

Vitamin A 0.2 g

Vitamin B1 0.25 g

Vitamin B2 0.3g

Water quantification

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 liter container, It is used in the production of the health beverage composition of the invention.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Formulation example  3: Manufacture of cosmetics

1. Preparation of Soft Cosmetics (Skin Lotion)

Combined extract of Bokbunja, Eoseongcho, Saenghwangwang and Hwapi 0.5%

Beta-1,3-Glucan 1.0%

Butylene Glycol 2.0%

Propylene Glycol 2.0%

Carboxy vinyl polymer 0.1%

Fiji-12 nonylphenyl ether 0.2%

Polysorbate 80 0.4%

Ethanol 10.0%

Triethanolamine 0.1%

Preservatives, colorings, flavors

Purified water to 100%

2. Nutritional Cosmetics ( Milk lotion Manufacturing

Combined extract of Bokbunja, Eoseongcho, Saenghwangwang and Hwapi 0.5%

Beta-1,3-Glucan 1.0%

Beeswax 4.0%

Polysorbate 60 1.5%

Sorbitan sesquioleate 1.5%

Liquid paraffin 0.5%

Caprylic / Capric Triglycerides 5.0%

Glycerin 3.0%

Butylene Glycol 3.0%

Propylene Glycol 3.0%

Carboxy vinyl polymer 0.1%

Triethanolamine 0.2%

Preservatives, colorings, flavors

Purified water to 100%

 3. Manufacture of nutritional cream

Bokbunja, Eoseongcho, Saenghwanghwang, Bokhwa extract 1.0%

Beta-1,3-Glucan 5.0%

Beeswax 10.0%

Polysorbate 60 1.5%

Sebum 60 Cured Castor Oil 2.0%

Solbitan Sesquioleate 0.5%

Liquid paraffin 10.0%

Squalane 5.0%

Caprylic / Capric Triglycerides 5.0%

Glycerin 5.0%

Butylene Glycol 3.0%

Propylene Glycol 3.0%

Triethanolamine 0.2%

Preservatives, colorings, flavors

Purified water to 100%

4. Agents for topical administration (gel ointment)

Complex extract of Bokbunja, Eoseongcho, Saenghwanghwang, Hwapi 33.0%

Beta-1,3-Glucan 10.0%

Polyacrylic Acid (Carbopol 940) 1.5%

Isopropanol 5.0%

Hexylene glycol 25.0%

Triethanolamine 1.7%

Deionized water to 100%

5. Agents for topical administration Patch Manufacturing

Bokbunja, Eoseongcho, Saenghwanghwang, Bokhwa extract 1.0%

Beta-1,3-Glucan 3.0%

Hexylene glycol 20.0%

Diethylamine 0.7%

Polyacrylic Acid (Carbopol 934P) 1.0%

Sodium sulfite 0.1%

Polyoxyethylene lauryl ether (E.O = 9) 1.0%

Polyhydroxyethylene cetyl stearyl ether (Cetomacrogol 1000) 1.0%

Viscous Paraffin Oil 2.5%

Caprylic Acid Ester / Capric Acid Ester Cetiol LC) 2.5%

Polyethylene Glycol 400 3.0%

Deionized water to 100%

Claims (10)

A pharmaceutical composition for the prevention or treatment of contact dermatitis or atopic dermatitis, comprising a composite extract containing Bokbunja, Eochocho, raw ground sulfur and Hwapi in a weight ratio of 1: 1: 1: 1. The composition of claim 1, wherein the complex extracts of Bokbunja, Eoseongcho, Saenghwanghwang and Hwapi are extracted with a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof. The composition of claim 2, wherein the complex extract is extracted with 60 to 85% (v / v%) of ethanol. delete delete The composition of claim 1, wherein the complex extract is contained in an amount of 0.001 to 50 wt% based on the total composition. Health functional food for the prevention or improvement of contact dermatitis or atopic dermatitis, comprising a complex extract containing Bokbunja, Eochocho, raw ground sulfur and hides in a weight ratio of 1: 1: 1: 1. delete delete delete

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