KR101068375B1 - Composition for the treatment of atopic dermatitis containing extracts of Saenghwanghwang, Seokchangpo, Red ginseng, Korean vinegar, and yellowish white as active ingredients - Google Patents

Abstract

The present invention relates to a composition for treating atopic dermatitis, which contains extracts of raw jihwang, Seokchangpo, red ginseng, licorice and yellow baek as active ingredients. More specifically, it can be used in various ways as a cosmetic composition, a pharmaceutical composition, and health functional food for treating atopy including the extract.

Description

Composition comprising extract from Rehmannia glutinosa Liboschitz, Acori Graminei Rhizoma, Sophora angustifola root, Lithospermi Radix and Phellodendron amurense bark for treating atopy}

The present invention relates to atopic dermatitis therapeutic composition containing the extracts of raw jihwang, Seokchangpo, ginseng, vinegar and sulfur white as an active ingredient, and more particularly, to a cosmetic composition, pharmaceutical composition, and health functional food for treating atopy including the extract.

Atopic dermatitis is a skin disease that recurs for a long time and shows itching and characteristic lesions. It is caused by various mechanisms such as genetic predisposition, environmental factors, skin barrier problems, pharmacological factors, psychological factors, and immunological factors. Most factors except skin barrier-related factors are difficult to control, so lesions may develop or worsen for a long time, and side effects from long-term treatment are difficult to avoid. Therefore, appropriate management is needed to prevent the recurrence or worsening of the lesion. The basic treatment of atopic dermatitis is skin moisturization, steroid external preparation, detection and removal of exacerbation factors, and education of patients and family, reflecting the importance of skin care. It can be said.

The stratum corneum is the most important barrier to protect our bodies from the outside. Its structure is a matrix of dead keratinocytes surrounded by membranes filled with keratin and filaggrin proteolytic products and mainly nonpolar lipids that fill the gaps, similar to bricking and cementing walls (Elias). PM.Stratum corneum defensive functions: an interated view.J Invest Dermatol 2005; 125: 183-200).

These intercellular lipids are composed of 50% ceramides, 25% cholesterol, and 10-20% long-chain free fatty acids to form multiple layers of lamellar sheets, which are permeable. It is essential to maintain barrier function normally.

Until recently, immune system abnormalities were considered to be the primary cause of atopic dermatitis, but evidence has been found that abnormalities in epidermal barrier function may provide a primary cause. Filaggrin is an important molecule in the formation of stratum corneum cytoplasm and filament coherence, while the degradation products play a central role in the hydration and acidification of the stratum corneum. The abnormalities have been found to be related to atopic dermatitis (Palmer CN, Irvine) AD, Terron-Kwiakowski A, Zhao Y, Liao H, Lee SP, et al. Common loss-of-function variants of the epidermal varrer protein filaggrin are a major prediposing factor for atopic deramtitis.Nat Genet 2006; 38: 441-446 ).

In addition, the secretion of the laminae, which has already mentioned the importance of maintaining the normal function of the barrier, is not normal in atopic dermatitis, resulting in a lack of intercellular lipids, especially ceramide components, of which type 3 ceramide is a transepidermal water loss. , TEWL). It is well known that atopic dermatitis also damages the antimicrobial barrier, leading to superinfection of Staphylococcus aureus, which in turn exacerbates dermatitis. Degradation of spingosine, a ceramide metabolite with a strong antimicrobial action as an antimicrobial barrier damage mechanism (Arikawa J, Ishivashi M, Karashima M, TAKAGI Y, Ichikawa G, Decreased levels of sphingosine, a natural antimicrobial agent, my be associated with vulnerability of the stratum corneum from patients with atopic dermatitis to colonization by Staphylococcus aureus.J Invest Dermatol. 2002; 119; 433-439), further damage of barrier function due to the release of ceramidases by microorganisms, skin Lack of antimicrobial peptides, such as LL-37, beta-defensins, and cathelicidin, produced in lamellar bodies and stored in keratosomes MH, Di Nardo A, Gallo RL.Keratinocytes store the antimicrobial peptide cathelicidin in lamellar bodies.J Invest Dermatol 2005; 124: 394-400). In addition, the chance of infection by viruses and fungi also increases. That is, in atopic dermatitis, water loss is caused by changes in the composition of filaggrin and intercellular lipids, which increases calcium ion concentration and cytokine in the epidermis, leading to proliferation of the epidermis, thickening and itching of the stratum corneum, and deterioration. Peptide deficiency increases the chance of infection by microorganisms. In addition, the deterioration of the protective membrane increases the chance of inducing an immune response by increasing the influx of external allergens into the skin. Therefore, skin care of atopic dermatitis should be focused on replenishing water loss due to deterioration of the protective film and eliminating infection opportunities.

At present, various studies have been conducted in the treatment of atopic dermatitis, but side effects and long-term recurrences of long-term use of therapeutic drugs are still typical problems. It is thought that suggesting an answer to this may be a convincing basis for the treatment of atopic dermatitis by using herbal medicine.

Skin plays a very important role as a barrier to protect individuals from the outside. Barrier function is to protect against various stimuli (chemicals, air pollutants, dry environment, ultraviolet rays, etc.) from the outside and to prevent excessive release of moisture through the skin. This protective function is a stratum corneum composed of keratinocytes. The function can be maintained only when is normally formed.

Transepidermal Water Loss (TEWL) is increased in normal-looking skin as well as in lesions and dry skin of patients with atopic dermatitis. The dry skin of AD patients is more closely related to the moisture content in the stratum corneum and the moisture entering through the stratum corneum rather than the normal skin. Moisture-binding capacity and water-holding capacity play an important role. In dry skin stratum corneum of AD patients, the water-binding capacity is not different from normal people, but water-binding capacity is reported to be decreased.

The outermost stratum corneum horner layer of the epidermis is formed from keratinocytes and consists of the differentiated keratinocytes and the surrounding lipid layer (Marcelo CL et al, J. Invest. Dermatol., 80 , pp 37-44, 1983). Keratinocytes are characteristic cells whose basal cells, which continuously proliferate in the epidermal layer, undergo morphological and functional changes in stages and rise to the surface of the skin. Exfoliated and new keratinocytes take over their functions, and this repetitive sequence of changes is called “epidermal differentiation” or “keratinization.” Also, keratinocytes are known to be naturally moisturizing factors. Natural moisturizing factor (NMF) and intercellular fats (ceramides, cholesterol, fatty acids) are produced, forming the stratum corneum, making the stratum corneum firm and flexible, retaining its function as a skin barrier.

 The stratum corneum can easily lose its function due to excessive face washing, lifestyle factors such as bathing, environmental factors such as dry air pollutants, and endogenous diseases such as atopic or senile skin. Recently, due to a variety of factors, more and more skin complaints and dry skin complaints are increasing in recent years.

Serum IgE is high in about 80-90% of patients, but serum IgE titer and severity of dermatitis are not necessarily proportional. There are two ways in which IgE autoresponse affects AD. First, autoallergens immediately induce allergic symptoms by cross-linking to applicator cells bound to IgE antibodies and releasing infectious media. This argument is supported by the discovery that IgE autoantigens release only hista from basophils and cause an immediate skin reaction (Meter C, 1999) (Crameri R, 1996). Second, it can be explained by the activation of the T-cell. In response, T-cells can be activated by monocute or dendritic cells through the appearance of IgE antibody mediated autoallergens. The presence of T-cells recognizing IgE antoantignes was indeed demonstrated in AD patients (Mossabeb R, 2002).

When the inflammatory reaction occurs, several proinflammatory mediators are produced, which are prostagrandin produced by nitric oxide (NO) and cyclooxygenase-2 (COX-2), which are produced by inducible nitric oxide synthase (iNOS). E ₂ (PGE ₂ ), and the like. These inflammatory factors activate the nuclera factor-κB (NF-κB), a transcription factor of the inflammatory response, resulting in excessive NO and PGE2 inflammation. In mammalian cell nitric oxide synthase (NOS), three isozymes exist: neuronal NOS (nNOS), endothelial NOS (eNOS), and inducible NOS (iNOS). Among these, iNOS is involved in the inflammatory response. In the case of iNOS, Interferon-γ. It is expressed in the presence of lipopolysaccharide (LPS) and various inflammatory cytokines. PGE2 is produced from Arachidonic acid by cyclooxygenase (COX). COX-2 is expressed in macrophage in response to infection or damage caused by microorganisms or stress of various factors. In other words, iNOS and COX-2 expression, NO and PGE ₂ production are representative inflammatory factors of immune cells. Inflammatory factors include tumor necrosis factor-α (TNF-α), interleulin-1β (IL-1β), and interleukin-6 (IL-6), which are inflammatory cytokines.

Therefore, various studies have been conducted to supply moisture from the outside to prevent proper skin moisture or to prevent moisture loss from the body, and various moisturizers having water retention capabilities have been developed. As a skin moisturizer, it is common to use lipid components such as ceramides and substances capable of increasing water retention in the stratum corneum such as essential fatty acids and lipid complexes (Rawlings AV et al, J. Invest. Dermatol., 5, pp731-). 741, 1994). However, in recent years, the risk factors for skin are gradually increasing, and the change of dietary habits slows down the formation and dropping rate of stratum corneum, and the function of keratinocytes decreases the amount of moisturizing factors and lipids in the stratum corneum, resulting in normal stratum corneum. There is a growing number of people with skin that do not function as a skin barrier. This disruption of skin barrier function causes various skin diseases such as dry skin, atopic dermatitis, contact dermatitis, and psoriasis. These diseases can be expected to alleviate symptoms only with a conventional moisturizer having a conventional water retention function, but fundamental healing is difficult.

In order to solve the existing problems, the present inventors have confirmed the effect as a composition for treating atopic dermatitis containing extracts of raw jihwang, changchangpo, ginseng, sorghum and baekbaek as active ingredients and completed the present invention.

An object of the present invention is to provide a cosmetic composition, a pharmaceutical composition, and a health functional food as a composition for treating atopic dermatitis containing extracts of raw jihwang, Seokchangpo, red ginseng, licorice and yellow white as active ingredients.

The present invention provides a composition for treating atopic dermatitis containing raw jihwang, Seokchangpo, Gosam, Jacho, and yellow baek extract as active ingredients.

In addition, the present invention provides a cosmetic composition comprising a composition containing raw jihwang, Seokchangpo, red ginseng, licorice, and baekbaek extract as an active ingredient.

In addition, the present invention provides a pharmaceutical composition comprising a composition containing raw jihwang, Seokchangpo, red ginseng, licorice, and baekbaek extract as an active ingredient.

In addition, the present invention provides a health functional food comprising a composition containing raw jihwang, Seokchangpo, Gosam, Licorice, and yellowish white extract as active ingredients.

The composition for the treatment of atopic dermatitis containing the raw jihwang, Seokchangpo, red ginseng, licorice, and baekbaek extract of the present invention as an active ingredient can be used in a variety of cosmetic compositions, pharmaceutical compositions and health functional food.

1 is a histological picture of the incision ear (NC / Nga mouse DNCB-induced dermatitis model- Nikon, 100 times the original size of Japan).
Figure 2 is a histological picture of the incision skin (NC / Nga mouse DNCB-induced dermatitis model- Nikon, 100 times the original size of Japan).
3 is a graph of scratching frequency over time for 7-8 subjects (mean ± SEM; P <0.05).
4 is a graph of scratching intensity over time for 7-8 subjects (mean ± SEM; a> b).
5 is a graph of the total IgE levels measured by ELISA in plasma (mean ± SEM; a> b).
Figure 6 is a graph of total IL-6 levels measured on animal cell culture media by ELISA (mean ± SEM).

Hereinafter, the components and technical features of the present invention will be described in more detail with reference to the following examples. However, the following examples are merely to illustrate the content of the present invention is not limited to the scope of the invention. The documents cited in the present invention are incorporated herein by reference.

Example

Example  1: Preparation of Sample

In order to achieve the above object, the acquisition of the medicines used in the present invention is as follows.

Where to buy Production Saenghwanghwang and Seokchangpo Namdo Herbal Agricultural Association Haengwon-ri, Jangheung-eup, Jangheung-gun, Jeonnam Gosam Chinese Medicine Love Inchon-ri, Bonghwa-gun, Gyeongbuk Self Chinese Medicine Love Fishing Village, Damyang, Chungbuk Yellow white Chinese Medicine Love Munchon-si, Gyeongbuk

The medicines were purchased in a dry state, stored frozen and ground and used. The organic solvents used in this study were grade 1 and special grade.

Each medicine was extracted repeatedly at room temperature with 70% alcohol. In other words, each of herbs such as raw jihwang, Seokchangpo, red ginseng, vinegar and yellow baek was repeatedly extracted three times by adding three times 70% alcohol solution per gram weight and soaking at room temperature for 3 days. Extracts of each medicine were used under reduced pressure filtration using a pore size 5㎛ paper filter paper (Advantec, No. 5).

For later experiments, the atopy solution was prepared by mixing the ratios of sulfuric acid: yellowish white: Seokchangpo: red ginseng: licorice in a ratio of 3: 2: 1: 1: 1. In addition, the cosmetic extract (cream) was prepared to be 10% of the total capacity of the mixed extract for animal experiments. The specific formulation is as follows.

Formulation example  1. Nutrition Cream Raw material Example 1 One Extract Mixture of Raw Yellow Sulfur, Seokchangpo, Red Ginseng, Licorice and Yellowish White  10 2 glycerin 1-4 3 Betaine 0.1 ~ 2 4 Caprylic / Capricglycerides 3 ~ 7 5 Jojoba Seed Oil 3 ~ 7 6 Camellia oil 2 to 4 7 Dimethicone 0.5 ~ 2 8 Cyclopentasiloxane 0.5 ~ 2 9 Tocopheryl acetate 0.1 ~ 2 10 Sorbitan tristearate 0.1 ~ 2 11 Sorbitan olivate 1-3 12 Hydroxyethyl cellulose 0.1 ~ 2 13 Sodium hyaluronate 0.5 ~ 3 14 Disodium ID 0.1 ~ 2 15 Allantoin 0.1 ~ 2 16 Sodium Polyacrylate 0.1 ~ 2 17 Rose Mario oil 0.1 ~ 2 18 Purified water Balance system 100.00

Hereinafter, the formulation examples of the pharmaceutical composition containing the extract of raw jihwang, Seokchangpo, Gosam, Jacho and yellowish white of the present invention will be described.

Formulation example  2. Powder Raw material Example 2 One Extracts of Raw Yellow Sulfur, Seokchangpo, Red Ginseng, Licorice and Yellowish White (Example 1) 300 mg 2 Lactose 100 mg 3 Talc 10 mg

Formulation example  3. Tablet Raw material Example 3 One Extracts of Raw Yellow Sulfur, Seokchangpo, Red Ginseng, Licorice and Yellow White (Example 1) 300 mg 2 Corn starch 100 mg 3 Lactose 100 mg 4 Magnesium Stearate 2 mg

Formulation example  4. Capsule Raw material Example 4 One Extracts of Raw Yellow Sulfur, Seokchangpo, Red Ginseng, Licorice and Yellow White (Example 1) 300 mg 2 Corn starch 100 mg 3 Lactose 100 mg 4 Magnesium Stearate 2 mg

Formulation example  5. Injection Raw material Example 5 One Extracts of Raw Yellow Sulfur, Seokchangpo, Red Ginseng, Licorice and Yellow White (Example 1) 300 mg 2 Sterile Distilled Water for Injection Quantity 3 pH regulator Quantity

Hereinafter, the formulation of the health functional food containing the extract of raw jihwang, Seokchangpo, Gosam, Jacho and yellowish white of the present invention will be described.

Formulation example  6. Healthy food Raw material Example 6 One Extracts of Raw Yellow Sulfur, Seokchangpo, Red Ginseng, Licorice and Yellow White (Example 1) 1000 mg 2 Vitamin mixtures Quantity 3 Vitamin A Acetate 1.0 mg 4 Vitamin E 0.13 mg 5 Vitamin B1 0.15 mg 6 Vitamin B2 0.5 mg 7 Vitamin B6 0.2 ug 8 Vitamin B12 10 mg 9 Vitamin c 10 ug 10 Biotin 1.7 mg 11 Nicotinic acid amide 50 ug 12 Folic acid 0.5 mg 13 Calcium Pantothenate Quantity 14 Mineral mixture Quantity 15 Ferrous sulfate 1.75 mg 16 Zinc oxide 0.82 mg 17 Magnesium carbonate 25.3 mg 18 Potassium phosphate monobasic 15 mg 19 Dibasic Potassium Phosphate 55 mg 20 Potassium citrate 90 mg 21 Calcium carbonate 100 mg 22 Magnesium chloride 24.8 mg

Formulation example  7. Healthy drink Raw material Example 7 One Extracts of Raw Yellow Sulfur, Seokchangpo, Red Ginseng, Licorice and Yellow White (Example 1) 1000 mg 2 Citric acid 1000 mg 3 oligosaccharide 100 g 4 Plum concentrate 2 g 5 Taurine 1 g 6 Purified water Quantity

Example  2: Atopic Inhibitory Effects in Animal Models

2-1. Preparation of the ingredients

After removing the back of the 11-week-old NC / Nga mice, they were left for 24 hours to heal fine wounds on the skin. Then 200 μL of 1% DNCB solution (acetone: olive oil = 3: 1) was applied to the back area, and after 4 days, 150 μL of DNCB solution was applied to the back area 2-3 times a week (from 12 to 16 weeks). . After 4 weeks of treatment, dermatitis was sufficiently induced, and all of the back skin was peeled off, and new dermatitis was formed on the site, so that the scratching action was intensified. DNCB treatment was stopped, and sensory evaluation was performed. 11-week old BALB / c mice were used as a normal group.

The dermatitis-induced mice were divided into two control groups (without atopy solution) and experimental group (20% atopy solution), and the two creams were applied for 5 weeks (from 12 to 17 weeks). After the sensory evaluation before and after drug treatment, blood was taken, and skin and lymph from the back and left ear were excised and stored in 10% formalin solution.

2-2. Histological evaluation

The skin tissue was examined to see how the atopy solution affected the skin tissue. Skin tissues were observed mainly on the overall condition of the skin, infiltration and degranulation of mast cells, and eosinophil infiltration. One of the characteristics of atopic dermatitis is that immune cells such as T cells, mast cells and eosinophils are infiltrated into the skin, and the epidermis and dermis become thick and bleeding occurs. Therefore, tissue staining was performed to confirm whether the atopy solution alleviated the symptoms of atopic dermatitis by preventing the infiltration of immune cells into the skin tissue.

Skin tissue immobilized in 10% formalin was embedded in paraffin, cut into 4um thickness, and stained with hematoxylin-eosin. In order to examine the infiltration of inflammatory cells, mast cells were stained by toluidine blue staining, and tissues were stained using congo red staining to confirm eosinophils. The magnification of was observed.

As a result, in the hematoxylin-eosin staining, the regular arrangement of tissue cells such as epithelial cells of the control group was destroyed, but the regular arrangement of tissue cells such as epithelial cells of the experimental group was observed. Toluidine blue staining, and Congo red staining, which stained mast cells, inhibited the degranulation of mast cells and their cells infiltrating skin tissue by atopic induction.

1 and 2 illustrate the effects of herbal extracts in atopic dermal animal models on infiltration and degranulation of mast cells.

2-3. Pruritus Behavioral Effects

In the case of atopic dermatitis, which is considered an immune disease, many studies have reported an increase in serum IgE levels. Atopic patients also have severe itching (pruritus). Analysis and comparison results to confirm the degree of pruritus, which is one of the general clinical symptoms of atopic dermatitis, are shown in Table 10, FIG. 3 and FIG.

Scratch Pattern Analysis Total Scratch Count ^{1 )} Scratch strength ^{2 )} Control group 169.89 ± 20.93 95.71 ± 10.63 Atopy 106.53 ± 16.65 ^* 60.30 ± 16.44 ^*

Average scratching of mice for 5 weeks. (/ 60 min) n = 8 Scratching behaviors of mice in all groups were taken with a video camera for 60 minutes.

1) This result calculated one period at a time.

2) Scratch strength was measured by the standards previously shown.

As the atopy is induced in the present invention, the number of scratching was increased, and when comparing the results of Weeks 2 and 3, the number and frequency of scratching were significantly lower in the experimental group of the present invention than in the control group.

On the other hand, Takahashi's study showed that scratching caused by pruritus increased in NC / Nga mice, an animal model of atopic dermatitis, and that the number of scratches in skin lesions in NC mice increased more than in skin-free lesions in NC mice. Reported.

2-4. In plasma IgE Influence of

IgE is an antibody that plays an important role in promoting atopic dermatitis by activating mast cells and is generally increased in atopic patients.

At 12, 15, 18, and 21 weeks of application, approximately 100 μL of blood was collected from the mice's eyes using capillaries, followed by centrifugation at 6,500 rpm for 20 minutes, and 30 μL of serum was isolated. . Serum was taken and stored frozen at −70 ° C. until used for experiments. Serum IgE was measured by ELISA kit (K & D system). Antibodies were diluted in coating buffer and coated in microwells and then placed at 4 ° C. overnight. Each well was washed three times with a wash buffer solution and then 100 μL of serum (100-fold dilution) was dispensed. This was allowed to stand at room temperature for 1 hour, washed twice with a wash buffer solution, and then treated with 100 μL of antibody avidin-HRP conjugated. The TMB substrate was dispensed by 100 μL and left in the dark for 30 minutes, and then treated with 50 μL of stop solution, and the absorbance was measured at 450 nm of ELISA reader. After 21-week-old NC / Nga mice were anesthetized with ketamine hydrochloride, blood was drawn by cardiac puncture, and serum was isolated. Quantification was performed with an immunoassay kit (ELISA kit).

Plasma IgE was significantly decreased by 20% in 117.25 ± 6.47 ng / ml in normal group, 133.92 ± 7.59 ng / ml in control group and 106.58 ± 8.11 ng / ml in experimental group (P <0.05). IgE is an antibody that plays an important role in promoting atopic dermatitis by activating mast cells, and is generally increased in atopic patients. In the present invention, the total IgE content in plasma was increased in the atopic dermatitis control group compared to the normal group. However, the experimental group significantly decreased to the level of the normal group. It was found that the experimental group reduced the IgE content in the plasma. In other words, it is 20% lower than that of the control group, and the above results suggest that the experimental group is effective in lowering the level of the immune factor IgE, but is not direct but may help to alleviate the symptoms of atopy (Fig. 5). ).

2-5. Of skin cell culture fluid IL -6 impact

IL-6 is a cytokine produced from several types of epithelial cells, such as activated macrophages, endothelial cells, and fibroblasts, which act on both innate and adaptive immunity, is involved in the growth of B lymphocytes, and mediates an overactive immune response.

After epilating the back of NC / Nga mice, 1g of skin tissue was removed, washed with DMEM culture medium, chopped into fine pieces, and replaced with 10% FBS-DMEM. After separating the culture supernatant for 7 days, the IL-6 secretion of the culture was measured by an enzyme immunoassay kit.

In the invention of IL-6 production of skin cell culture medium, the increase in DNCB-induced atopic dermatitis was increased in the control group. (FIG. 6).

While the present invention has been described with reference to exemplary embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. You will know. In addition, many modifications may be made to adapt a particular situation and material to the teachings of the invention without departing from the essential scope thereof. Accordingly, it is intended that the invention not be limited to the particular embodiment disclosed as the best mode contemplated for carrying out this invention, but that the invention be construed as including all embodiments falling within the scope of the appended claims.

Claims (3)

A composition for improving atopic dermatitis comprising raw jihwang, Seokchangpo, red ginseng, licorice, and yellowish white extract as active ingredients. A pharmaceutical composition for the treatment of atopic dermatitis comprising raw kojihwang, Seokchangpo, red ginseng, licorice, and yellowish white extract as active ingredients. Health functional food for improving atopic dermatitis containing raw kojihwang, Seokchangpo, red ginseng, vinegar and yellow white extract as active ingredients.

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