KR20200015989A - Composition for the treatment of atopic dermatitis, which comprises an extract of Rumex japonicus Houtt - Google Patents

Abstract

The present invention relates to: a composition for treating and alleviating atopic dermatitis containing a Rumex japonicus extract; and a method for manufacturing the extract. The composition for treating and alleviating atopic dermatitis of the present invention uses the Rumex japonicus extract extracted by using an organic solvent, and has an excellent effect of treating atopic dermatitis and alleviating symptoms thereof.

Description

Composition for the treatment of atopic dermatitis, including the extract of Chameori scabbard {which comprises an extract of Rumex japonicus Houtt}

The present invention relates to a composition for the treatment and improvement of atopic dermatitis and extracts comprising the extract of Chamsori.

Atopic dermatitis is a multi-interactive skin disease with complex interactions. Various factors, including immunological abnormalities, contribute to the pathogenesis and onset of atopic dermatitis (Choi, et al., 2013). Atopic dermatitis, which occurs mainly in childhood, is the most common chronic skin disease and can affect the quality of an individual's life span (Choi, et al. 2012). Common symptoms of atopic dermatitis include dry and inflamed skin, intense itching, skin irritability, repeated rashes, persistent scratches, erythema and blisters, and chronic or severe sleep disorders resulting in insomnia, psychology And emotional pain, poor quality of life. (Papier and Strowd 2018) However, the exact etiology and effective treatment for atopic dermatitis has not yet been established. Thus, therapeutic agents for atopic dermatitis, which have been developed to date, are therefore anti-inflammatory agents such as steroids, tacrolimus, topical anti-inflammatory agents such as Pimecrolimus, antihistamines and cyclosporines (Lancet. 1991; 338: 137 -140), and hypoallergenic moisturizer as an adjuvant therapy, photochemotherapy with ultraviolet A (Photochemotherapy, J. Am. Acad. Dermatol. 1998; 38: 589-593, Lancet. 2001; 357: 2012 2016 is also being implemented 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.

Until recently, topical steroids have been the dominant drug for treating atopic dermatitis, but when used for a long time, various side effects occur, infections worsen due to immune mechanism disorders, and side effects that weaken skin resistance are used. When to stop there is a problem that causes a serious symptom called steroidal contact dermatitis. Along with this, emollients or oral antihistamines are used as the primary therapy for atopic dermatitis, but these drugs are also always a problem for long-term use (Choi, et al., 2012; -S., Et al. , 2006). To minimize these side effects, patients with atopic dermatitis may use herbal remedies (Gao, et al., 2005; Koo and Arain 1998). Therefore, the development of atopic dermatitis treatment without side effects is required.

Rumex japonicus Houtt. Is a perennial herb distributed throughout Japan, Korea, and China, which has traditionally been used as an antimicrobial, purifying, anti-inflammatory and anti-tumor agent in folk medicine (BELKIN and FITZGERALD 1952; Jiang, 2007; Zhou, et al 2005). Chassams are also known to contain large amounts of antioxidants such as anthraquinones, flavonoids and triterpenoids (Vasas, et al., 2015). In the previous study, EtOH extract of Rinnia root had a strong antioxidant activity (total polyphenol = 158 ± 6.3 μg / ml, DPPH radical scavenging activity = 71.31 ± 3.8%), which contains a large amount of phenolic compounds in the extract It was confirmed that it is.

In this regard, the present inventors have confirmed the anti-atopic effect of the extract of Sesame Japonica on TNF-α-induced responses in vitro and in vivo. In particular, the Sesame Japonica extract acts on the regulatory MAPK pathway and the NF-κB activation pathway. It was confirmed to have an atopy effect. In addition, the effect of the extract of Chamsori-rigi to reduce the development of atopic dermatitis was also confirmed. In particular, the effects on the solvent-specific extracts are different, and thus the present inventors have completed the present invention including the extraction method of the organic solvent extract having the best therapeutic effect against atopic dermatitis.

Patent Registration No. 10-1523554

An object of the present invention is to provide a pharmaceutical composition for the treatment or prevention of atopic dermatitis, including the extract of Chamsori root.

Another object of the present invention is to provide a cosmetic composition comprising the root extract of Chamsori, which has a skin irritation relief, promote regeneration, and further have a symptom relief effect associated with atopic dermatitis.

Still another object of the present invention is to provide a method of extracting the extract having the above effect from the root of Chamsori.

In order to achieve the object of the present invention as described above, the present invention provides a pharmaceutical composition for the treatment or prevention of atopic dermatitis, including the extract of Sesame root.

In one embodiment of the present invention, the extract of Roots of Chamsori is characterized by using an organic solvent except water, but is not limited thereto, methanol (methanol), ethanol (ethanol), propanol, isopropanol (isopropanol) ), Alcohol containing 1 to 4 carbon atoms including butanol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride , Characterized in that extracted with at least one organic solvent selected from the group consisting of hexane (cyclohexane). In one embodiment of the present invention, the extract was prepared from the roots of Dried Cham Suk Rim using ethanol. In the case of using an organic solvent, the active ingredient content in the extract is different from the water extract, and it was experimentally confirmed that the treatment and amelioration effect for atopic dermatitis was superior.

The extract of the root of Chamsori of the present invention may be included in the pharmaceutical composition at a concentration of 1 to 200 µg / ml, and more preferably 10 to 50 µg / ml. When the concentration of the extract is less than 1 μg / ml, the effects of treatment and improvement on atopic dermatitis symptoms are insignificant, and the effects are difficult to measure, and when the concentration is 200 μg / ml or more, the therapeutic effect may be lowered.

 The pharmaceutical compositions of the present invention also have the effect of inhibiting the MAPK and NF-κB signaling pathways, which in turn have the effect of treating and treating symptoms for atopic dermatitis. From the results of inflammation-induced cell experiments, in particular, the effects of participating in and inhibiting the signal transduction pathways were confirmed through experiments. In addition, clinical therapeutic effects were confirmed through animal experiments.

In another aspect of the present invention, the pharmaceutical composition of the present invention is characterized in that it is applied topically to the affected area as an external preparation for skin. Although there is no limitation in the application method, it has been confirmed that the clinical effect is excellent as a topical application rather than a systemic oral administration. Especially, when used as an external skin supplement with oral administration of other drugs, the therapeutic effect against atopic dermatitis is excellent. Do.

In addition, in order to achieve the other object of the present invention, the present invention provides a cosmetic composition comprising the root extract. The cosmetic composition does not have a therapeutic effect due to a slight effect on the human body, but is a functional cosmetic composition that can alleviate the symptoms of atopic dermatitis due to effects such as skin irritation relief and skin regeneration promotion.

Finally, in order to achieve another object of the present invention, the present invention provides a method for preparing the extract of Chamsori. It has been experimentally confirmed that the extract of Chassori root, which is prepared according to the method of extract of the present invention, has a very good treatment or improvement effect of atopic dermatitis symptoms, in particular, due to the differentiation of the solvent.

The present invention relates to a pharmaceutical composition for the treatment or prevention of atopic dermatitis, including the extract of Chamsori root, a cosmetic composition for alleviating the symptoms of atopic dermatitis, and a method for preparing the extract. Excellent treatment and prevention of atopic dermatitis, symptom relief effect.

Figure 1 shows the growth rate of the cells treated with Sesame root root extract, Figure 1A is treated with HaCaT cells of various concentrations of Sesame root root extract, after measuring the growth rate over time, Figure 1B is in TNF-α By measuring the effect of the extract of the roots of the Koreans in the inflammatory response induced by the cell.
Figure 2 is the result of measuring the expression level of the protein induced by TNF-α in HaCaT cells.
Figure 3 is a result of measuring the effect of the root extract of sangryokja on TNF-α-induced protein expression in HaCaT cells, Figure 3A shows the expression of ERK1 / 2, p38 induced in the MAPK signaling pathway, 3B shows the effect on Akt induced by stimulation of TNF-α, and FIG. 3C shows the effect on the NF-κB signaling pathway.
4 is a diagram schematically illustrating an experimental design for an animal model causing atopic dermatitis.
FIG. 5 shows the results of experiments comparing the effects of erythema, dryness, itching, edema, blisters, thyroiditis, etc., and clinical treatments of the root extract of Ames dermatitis.
Figure 6 shows the results of experiments comparing the effect of the ear thickness of the mouse induced atopic dermatitis, and the treatment of the root extract.
7 and 8 show the result of comparing the weight of the visceral organs of mice that induced atopic dermatitis.
9 and 10 show the results of histological changes in the back skin and ears of the mouse induced atopic dermatitis.
Figure 11 shows the results of comparing the polyphenol content in the extract of the root of chamsori (ethanol extract) and hot water extract of the present invention.
Figure 12 shows the results of comparing the radical scavenging ability from the DPPH measurement results of the chamsori root extract (ethanol extract) and hot water extract of the present invention.

The present invention relates to the use for the treatment and prevention of atopic dermatitis, etc. of the root extract of Chamsori, the extract can be utilized as a pharmaceutical composition or cosmetic composition.

As mentioned above, the extract of Roots of Chamsori is characterized by being extracted from an organic solvent, including methanol, ethanol, propanol, isopropanol, butanol, and the like. Alcohols with 1 to 4 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane and cyclohexane It can be extracted using at least one organic solvent selected from the group consisting of).

In addition, the extraction method may be used by selecting any one of cold extraction, reflux cooling extraction, solvent extraction, steam distillation, ultrasonic extraction, elution, compression method. In addition, the desired extract may further be subjected to a conventional fractionation process, and may be purified using conventional purification methods. There is no restriction | limiting in particular in the manufacturing method of the extract of this invention, Any known method can be used. Therefore, in the present invention, the chamsori root extract is a concept that includes all the extracts, fractions and purified products obtained in each step of extraction, fractions or purification, their dilutions, concentrates or dried products.

More specifically, in one embodiment of the present invention was extracted three times using a cold needle extraction method using ethanol. More specifically, washing and pulverizing dried Chaksam root; Cold extraction was performed three times at 25 ° C using 95% ethanol. The extract obtained was then freeze-dried to obtain the extract of the present invention.

The composition of the present invention is a pharmaceutical composition comprising the extract of the root of Chamsori as an active ingredient can be prepared using a pharmaceutically acceptable and physiologically acceptable adjuvant in addition to such an active ingredient, the adjuvant, boron Releases, sweeteners, binders, coatings, swelling agents, lubricants, lubricants, flavoring agents, and the like can be used. The pharmaceutical composition may be preferably formulated into a pharmaceutical composition including one or more pharmaceutically acceptable carriers in addition to the above-described active ingredient for administration.

Formulation forms of the pharmaceutical compositions may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, nontoxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrating agents and coloring agents may also be included in the mixture. Suitable binders include, but are not limited to, natural and synthetic gums such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as 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 in compositions formulated in liquid solutions are sterile and biocompatible, which include saline, sterile water, Ringer's solution, buffered saline, albumin injectable solutions, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers and bacteriostatic agents may be added as necessary. Diluents, dispersants, surfactants, binders and lubricants may also be added in addition to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA can be formulated according to each disease or component according to the appropriate method in the art.

In particular, the pharmaceutical composition of the present invention may be preferably formulated into a topical application to the skin, but is not limited thereto, and may be a formulation such as an ointment, a spray or a patch. When formulated for sprays such as aerosols, a propellant or the like may be combined with the additives to disperse the dispersed dispersion or wet powder. When the pharmaceutical composition is an ointment, an ointment base such as an oil-based base, a water-soluble base, and an emulsion base, an antioxidant, an antiseptic, a moisturizing agent, and a dissolution aid may be included.

Chamsori root extract of the present invention may be formulated in the form of cosmetics. When formulated in cosmetics, it may include ingredients conventionally used in cosmetic compositions, such as conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavors, and carriers. In addition, the composition of the present invention may be used in addition to the above-mentioned Chaksam Ryukju root extract, atopic dermatitis inhibitors that have been conventionally used in a range that does not impair the skin protection effect by reacting with Chaksam Risa root extract.

Examples of products to which the cosmetic composition of the present invention may be added include, for example, cosmetics such as astringent cosmetics, soft cosmetics, nourishing cosmetics, various creams, essences, packs, foundations, and the like, cleansing agents, face washes, soaps, treatments, and cosmetics. Etc.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited to these examples.

< Example  1> Experimental Materials and Methods

1-1. Experimental material

Bovine serum albumin (BSA), Dulbecco's modified Eagle's Medium (DMEM), Fetal bovine serum (FBS), penicillin, streptomycin and trypsin were purchased from Gibco-BRL (Grand Island, NY, USA). Dimethylsulfoxide (DMSO), 3- (4,5-dimethyl thiazol-2-yl) -2,5 diphenyltetrazolium bromide (MTT) was manufactured by Sigma-Aldrich Inc. (St. Louis, MO) Was obtained and used. Phospho-Akt (Ser473), phospho-ERK, phospho-p38, phosphor-IκBα, NF-κB p65, Lamin B1 and GAPDH antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA). TNF-α was purchased from R & D systems (Tokyo, Japan). All other reagents used used the most appropriate grade.

1-2. Preparation of Sesame Root Extract

The dried roots of the Rumex japonicus Houtt were provided by Keratin Korea (Busan, Korea). The roots were washed three times to remove impurities, dried at room temperature and stored at 4 ° C. until use. The dried roots were ground with a grinder and extracted three times with 95% ethanol for 3 days at 25 ° C., and then the extract was filtered through Advantech No. 3 filter paper (Osaka, Japan). The filtered liquid was evaporated using a rotary vacuum evaporator (Tokyo Rikakikai Co., Ltd, Tokyo, Japan). The final product obtained was lyophilized in vacuo.

1-3. Cell culture and cell viability measurement

Cell viability was measured by the MTT assay described above. HaCaT human keratinocyte cells were incubated at 37 ° C., 5% CO ₂ conditions in DMEM containing 10% FBS and 100 μg / ml penicillin-streptomycin. HaCaT cells were plated at a density of ⁴ × 10 ⁴ cells / well in 24-well plates and incubated over night in growth DMEM medium. Cells were washed with fresh culture to remove cells, and the extracts from the previous stage were treated with various concentrations and cultured for 24 hours, 48 hours and 72 hours. MTT solution (5 mg / ml) was added to each well and incubated for 3 hours at 37 ° C. Finally, DMSO was added to dissolve the formazan salt and the amount of formazan produced was measured using an GENios microplate spectrophotometer (PowerWaveTMXS, BioTek Instruments, Inc., Winooski, USA) for optical density at 540 nm (OD). Was measured.

1-4. Western blotting analysis

Cells were incubated at a density of 5 × 10 ⁴ cell / well in 6-well plates for 24 hours to incorporate DMEM. After the adaptation process, the cells were treated for 30 minutes with the extract of Rinse root, and then stimulated with TNF-α (10 ng / mL) for 30 minutes in serum-free medium. The cells were washed with cold PBS and then scraped and treated with 300 μL of RIPA buffer containing a protease inhibitor (TransLab, Daejeon, Korea). The lysate was separated by 10% SDS-polyacrylamide gel and then transferred to PVDF membrane (Bio-Rad, CA USA). Western blot was probed with a specific antibody at 4 ℃, over night. After incubation with horseradish peroxidase-conjugated secondary antibody (Bethyl Laboratories Inc., Montgomery, USA) for 1 hour at room temperature. This was visualized using an enhanced chemiluminescence method (ECL), Amersham Biosciences, Buckinghamshire, UK and analyzed by ChemiDoc XRS (Bio-Rad, CA). Densitometer analysis was performed using a Hewlett-Packard scanner and Image Lab software.

1-5. Experimental animals

Five-week-old female Balb / c mice were purchased from Samtako Inc. (Osan, Korea) and bred at the experimental animal research center of Gyeongsang National University. Mice were housed in cages under standard conditions, freely fed food / water and bred in a 12 hour light cycle. The room temperature was 23 ± 2 ° C. and the humidity was 35 to 60%. The animal testing protocol used in this study was approved by the Gyeongsang National University Animal Testing and Use Council. The animal testing protocol number is GNU-160623-M0015.

1-6. Experimental Study of Chamsori Root Extract on Atopic Dermatitis

Experimental mice were divided into 5 groups (n = 5 per group). DNCB was applied to the back skin and ears to induce immune responses and skin lesions caused by atopic dermatitis. After complete removal of the dorsal skin of the mice, 200 μL of 0.5% DNCB solution (dissolved in acetone and olive oil 3: 1 mixture) was treated on days 1 to 3 of the experiment. On days 8 to 35, 20% of 1% DNCB and 200μL were applied three times per week to induce atopic dermatitis, followed by topical application of Sesame root extract to the skin and ears of mice for 3 weeks daily. As a control, Dexamethasone was applied to mice with a 1.5 mg / kg I.P injection three times per week. On day 35 of the experiment, the experimental animals were sacrificed.

1-7. Atopic Dermatitis Lesion Evaluation

The severity of dermatitis was evaluated by the method proposed by Yamamoto, et al . The severity of dermatitis was assessed once a week. The degree of erythema / bleeding, scar / dry, edema, threshing / corrosion was evaluated as 0 (none), 1 (slight), 2 (moderate), 3 (severe). The sum of the individual list scores was taken as the dermatitis score.

1-8. Ear thickness and organ weight measurements

Ear thickness was measured by micrometer on the day of sacrifice. Micrometers were recorded and measured (μm) near the tip of the ear next to the cartilage ridge. Lymph nodes and spleen weights were measured on an electronic balance, respectively.

1-9. Pathological research

The skin and ear lesions were thinly sliced and the tissue sections fixed in 10% buffered neutral formalin for 24 hours. Fixed tissue sections were embedded in paraffin wax, sectioned, deparaffinized and rehydrated using standard techniques. Sections 5 μm thick were subjected to hematoxylin and eosin (H & E) staining and toluidine blue staining for detection of various inflammatory cells, and histopathological changes were observed under an optical microscope. Each microscope field observed an arbitrary range at magnifications of x100 (skin) and x200 (ear).

1-10. Statistical analysis

The results are expressed as mean ± standard deviation (S.D.). One-way ANOVA was used to assess the importance of the difference between the two mean values.

* P <0.05 and ** P <0.01 were considered statistically significant.

Example 2 Experimental Results

2-1. Effect of Black Root Extract on the Survival Rate of HaCat Cells

MTT analysis was performed to determine the effect of Rhizome root extract in HaCaT cells. HaCaT cells were incubated for 24, 48 and 72 hours at various concentrations of RJ. As a result, when treated with the extract of the roots of the present invention, the cell viability showed a tendency to increase most (Fig. 1A), the roots extract did not have cytotoxicity at all concentrations. Chamsori root extract was used at concentrations of 25 and 50 μg / ml in future experiments. In addition, the cytotoxic effect of the Black Root Extract on HaCaT cells in the presence of TNF-α was measured. As a result, as shown in FIG. 1B, the extract of Roots of Roots of Cerrus was not cytotoxic up to a concentration of 50 μg / ml in HaCaT cells stimulated with TNF-α. This result suggests that the extract of Chamsori root may have anti-atopic dermatitis effect through HaCaT cells.

2-2. Effect of Black Root Extract on TNF-α Induced MAPK Family and NF-κB Pathway

Next, the phosphorylation of p-38, ERK and Akt was investigated in TNF-α stimulated in HaCaT cells. p-p38 levels increased from 30 minutes to 180 minutes with TNF-α treatment, and phospho-ERK and phospho-Akt levels increased after 30 minutes. TNF-α was treated for 30 minutes for further experiments, which induced a substantial inflammatory response to TNF-α increase in p38, erk and akt expression. (FIG. 2).

Cells stimulated by TNF-α increased the expression of p-p38 and p-erk. However, when treated with the root extract of Chamsori, p-p38 and p-erk significantly decreased (Fig. 3A). In addition, the effects of the Black Root Extract on the NF-κB pathway were evaluated. As shown by Western blot analysis, it was found that Akt was overexpressed in HaCaT cells treated with TNF-α. However, when treated with the root extract of Chaksam, significantly reduced the phosphorylation induced by TNF-α of Akt (Fig. 3B). In the unstimulated state, NF-κB is maintained in the cytoplasm as an inactive complex with IκBα, an inhibitory protein that blocks the nuclear importing sequence of NF-κB. Transcriptional activation of NF-κB is regulated by phosphorylation at serine 536 residues in the transcriptional activation domain of the functionally active subunit p65 / RelA (Kalra, et al., 2008). We have confirmed the effects of the oleander root extract on the activation of NF-κB induced by TNF-α. As a result, the Rhizome root extract dose-dependently inhibited the phosphorylation of IκBα and nuclear transfer stimulated by TNF-α of NF-κB p65 (FIG. 3C).

2-3. Anti-atopic Effects of Extracts from the Roots of Roots in Vivo

From the results demonstrating that Creeper Root Extract can effectively reduce inflammation in HaCaT cells, we used Balb / c mice to investigate the role of Creeper Root Extract in anti-atopic efficacy. Mice that induced atopic dermatitis were topically treated with DNCB, and then applied using dexamethasone as a root control extract and a positive control as described in FIG. 4. Application of DNCB for 1 week resulted in significant inflammation including erythema, vesicles, edema, scratches, moss and dry skin. Subsequently, after treatment by applying the root extract of the ulsori to atopic dermatitis mice, it showed a reduced dermatitis score (Fig. 5). On the other hand, the dermatitis score of mice treated with dexamethasone was higher than that of Roots of Roots. As a result similar to the dermatitis score, reduced ear thickness was observed in a dose-dependent manner in the control group of the Blackberry root extract treatment group (FIG. 6). This result demonstrates that the Black Root Extract reduces atopic symptoms in Balb / c mice.

2-4. Lymph Node and Spleen Weight Reduction Effect of Korean Radix Root Extract

Lymph nodes and spleen weights were measured to determine whether topical administration of the extract of Rhizome root induced anti-atopic effects in mice. The control group had increased lymph node and spleen weight in mice compared to the sham group. Group of the Black Root Extracts were dose dependently inhibited in lymph node weight (FIG. 7). In addition, the spleen weight was slightly decreased after treatment with the Black Root Extract (FIG. 8). Taken together, this shows that topically applied Chamsori root extract can have an anti-atopic effect.

2-5. Effect of Rhizome Root Extract on Atopic Dermatitis-like Lesion Mice

To investigate the association between in vivo and in vitro experiments, we confirmed the effect of topical application of the extract of Rhizome root on DNCB-induced mice. Atopic dermatitis is known to cause increased skin thickness and excessive lymphocyte infiltration into the dermis. Skin and ears were stained with hematoxylin & eosin and toluidine blue and observed with light microscopy. Observations through H & E staining showed that the control group significantly increased the thickness of epidermal and dermal tissues, whereas the Chassoris root extract treatment group significantly alleviated these changes, especially in epidermal tissues compared to the control. In addition, toluidine blue staining was noticeable in the number of mast cells in the dermal area, while the Rhizome root extract treatment group reduced the number of mast cells depending on the dose administered (FIGS. 9 and 10).

Example 3 Comparison of Root Water Extract and Ethanol Extract

1. Polyphenol Content Comparison

In order to compare the polyphenol content of the ethanol extract of Chamsoris roots prepared in Example 1 of the present invention, and the hot water extracts of Chamsoris roots, the ethanol extracts, hydrothermal extracts, and garlic of the roots of Chamsoris extracts extracted by the extraction method of the present invention. Acids were prepared by diluting to 200, 100, 50, 25, 12.5, 6.25, 3.125 and 0 μg / ml, respectively. 10 μl of each extract sample and gallic acid were dispensed into a 96 well plate, and 50 μl of folin-ciocalteu reagent was added to the foil, followed by shaking for 5 minutes. Next, after adding 40 μl 7.5% Na ₂ CO ₃ , it was wrapped in foil and shaken for 2 hours. The OD value of each sample was measured at 750 nm.

As a result, it was measured that 33.6 µg / mg of the ethanol extract of the root of Chamsori, and 26.2 µg / mg of the hot water extract contained polyphenols. In the case of the ethanol extracts of the root of Chamsori, the polyphenol content was determined to be about 1.3 times higher than that of the hydrothermal extract. (Figure 11)

2. DPPH Analysis

In order to compare the radical scavenging ability of the ethanol extract and the hot water extract of the root of Chamsori-ri, extracted by the extraction method of the present invention, DPPH analysis of each extract was performed. Each extract sample was prepared at a concentration of 2.5 mg / ml, 5 mg / ml, 10 mg / ml, and the DPPH stock solution was diluted in DMSO (DPPH MW: 394.2 g / L = 0.0118 g / ml, then diluted 50-fold, 200 μl DPPH + 9.8 ml DMSO).

450 μl of the DPPH solution was added to the e-tube, and 50 μl of the extract sample was added thereto. A control was prepared by adding the same amount of extract to 450 μl of DMSO. 100 μl of the prepared sample was dispensed on a 96 plate and shaken for 1 hour. The OD values of the prepared samples were measured at 517 nm and compared. (Radical scavenger ratio: (Control- (sample-sample blank)) / control * 100)

As a result of comparing the measured values, it was confirmed that the radical scavenging ability of each extract was excellent in ethanol extract, especially 71.8% at the concentration of 5 mg / ml of ethanol extract, and 10 mg of hot water extract. The highest was 27.6% at / ml. (Figure 12)

So far I looked at the center of the preferred embodiment for the present invention. Those skilled in the art will appreciate that the present invention can be implemented in a modified form without departing from the essential features of the present invention. Therefore, the disclosed embodiments should be considered in descriptive sense only and not for purposes of limitation. The scope of the present invention is shown in the appended claims rather than the foregoing description, and all differences within the scope will be construed as being included in the present invention.

Claims (12)

A pharmaceutical composition for the treatment or prevention of atopic dermatitis, including the extract of Chaksam. The method of claim 1,
The extract of Roots of Chamsori is alcohol having 1 to 4 carbon atoms, including a methanol, ethanol, ethanol, propanol, isopropanol, butanol, butanol, acetone, acetone, ether , Extracted using at least one organic solvent selected from the group consisting of benzene, chloroform, ethyl acetate, methylene chloride, hexane, and cyclohexane. Pharmaceutical composition for treating or preventing atopic dermatitis. The method of claim 2,
The Chamsori root extract is a pharmaceutical composition for treating or preventing atopic dermatitis, characterized in that extracted using ethanol (ethanol). The method of claim 1,
The extract is a pharmaceutical composition for treating or preventing atopic dermatitis, characterized in that the composition is contained in a concentration of 1 to 200 ㎍ / ml. The method of claim 1,
The extract of Roots of Chamsori Jeryong, characterized in that it has the effect of inhibiting the MAPK and NF-κB signal transduction pathways for the treatment or prevention of atopic dermatitis. The method of claim 1,
The pharmaceutical composition for treating or preventing atopic dermatitis, characterized in that the external application for the skin. Cosmetic composition comprising the root extract. The method of claim 7, wherein
The cosmetic composition is a cosmetic composition, characterized in that having a skin irritation relief or skin regeneration promoting effect. The method of claim 7, wherein
The cosmetic composition is a cosmetic composition, characterized in that having atopic dermatitis symptomatic effect. Washing and drying the dried sesame root;
Extracting at room temperature by adding an organic solvent to the root of Chamsori; And
The freezing and drying step of extracting the root of the extract, Chamsori root extract. The method of claim 10,
The organic solvent is methanol (methanol), ethanol (ethanol), propanol (propanol), isopropanol (isopropanol) and butanol (butanol) is a method for producing a root extract of Chamsori, characterized in that at least one selected from the group consisting of. The method of claim 10,
The chassam root extract is characterized in that it has a treatment or amelioration effect of atopic dermatitis symptoms, the method of producing Chamsori root extract.

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