KR102149851B1 - Novel Use of Sodium Surfactin for Improvement of Inflammatory Skin Diseases - Google Patents

Abstract

The present invention relates to a novel use of sodium surfactin, and more particularly, to a use of sodium surfactin for treatment/improvement of inflammatory skin diseases or skin moisturizing.
Sodium surfactin according to the present invention not only concentration-dependently reduces the expression of NO induced by irritants or IL-6, an inflammatory cytokine, but also skin dryness, eczema, itching, erythema in mouse models or actual human application tests. It has an alleviating effect, and also provides a skin barrier strengthening effect and a skin moisturizing effect, and does not show cytotoxicity, so it is also useful as a good treatment strategy for inflammatory skin diseases that do not cause side effects.

Description

Novel Use of Sodium Surfactin for Improvement of Inflammatory Skin Diseases

The present invention relates to a novel use of sodium surfactin, and more particularly, to a use of sodium surfactin for treatment/improvement of inflammatory skin diseases or skin moisturizing.

Sodium Surfactin is a peptide lipid composed of amino acids and fatty acids, and is produced from fermentation of Bacillus subtilis. It is known as a biological surfactant and is environmentally friendly because it is easily biodegradable. Biological surfactants can be applied to food, detergent, pulp and paper, crude oil, pharmaceuticals, cosmetics, and environmental purification. In addition, Korean Patent Laid-Open Publication No. 10-2004-0055035 discloses a method for preparing sodium surfactin and an inhibitory effect on collagen degrading enzymes using the prepared cerdium surfactin, but an anti-inflammatory effect, particularly a use related to inflammatory skin diseases, has been reported. none.

Meanwhile, among the recent living environments, harmful factors to humans are gradually increasing, and the elderly population is rapidly increasing. Accordingly, the rate of formation and drop-out of the stratum corneum is slowed, the ability of keratinocytes to synthesize lipids decreases, or the amount of moisturizing factors and lipids in the stratum corneum is reduced due to the inability to divide, mature and differentiate normal cells from the epidermis. There is also an increasing trend of people who have skin that cannot maintain the normal stratum corneum function, that is, the skin barrier function is broken.

Due to the division and differentiation of such abnormal epidermal cells, the skin causes various inflammatory skin diseases such as dry skin, atopy, and psoriasis. These diseases can only slightly alleviate the symptoms only with conventional moisturizers that have only the moisture retention function on their own. In other words, it is difficult to expect that fundamental healing. Therefore, it is time to develop a composition for treating and improving inflammatory skin diseases such as skin atopy, dry skin, and eczema. Therefore, therapeutics or functional cosmetics are being developed to improve inflammatory skin diseases such as allergic dermatitis or atopic dermatitis, but there are cases where the effect of improving inflammatory skin diseases is insignificant or toxicity or side effects are concerned by using the compound as an active ingredient. Development of new materials derived from natural products was requested.

Accordingly, the inventors of the present invention have made diligent efforts to develop a new inflammatory skin disease treatment or cosmetic composition that can effectively work on inflammatory skin diseases and solve the side effects of existing drugs or functional cosmetics.As a result, sodium surfactin, a natural material, has an anti-inflammatory effect. While it was confirmed that it has a moisturizing effect to improve skin itching and dry skin, the present invention was completed.

The information described in the background section is only for improving an understanding of the background of the present invention, and thus does not include information forming the prior art known to those of ordinary skill in the art to which the present invention belongs. May not.

Patent Document 1: KR10-2004-0055035 A

It is an object of the present invention to provide a novel and novel use of sodium surfactin.

Another object of the present invention is to provide a composition for preventing or treating/improving inflammatory skin diseases.

Another object of the present invention is to provide a natural material that improves skin inflammation and itchiness, dry skin, eczema, etc. without side effects and provides excellent moisturizing effects.

In order to achieve the above object, the present invention provides a novel use of sodium surfactin.

Specifically, the present invention provides a pharmaceutical composition for the prevention or treatment of inflammatory skin diseases containing sodium surfactin as an active ingredient.

The present invention also provides a cosmetic composition for improving inflammatory skin diseases or moisturizing skin, containing sodium surfactin as an active ingredient.

The present invention also provides a functional cosmetic for improving inflammatory skin diseases containing sodium surfactin as an active ingredient.

The present invention provides a novel composition for preventing or treating inflammatory skin diseases and a cosmetic composition for improving inflammatory skin diseases capable of specifically and effectively treating/improving inflammatory skin diseases without problems of toxicity or side effects of existing drugs or functional cosmetics.

Sodium surfactin according to the present invention not only concentration-dependently reduces the expression of NO induced by irritants or IL-6, an inflammatory cytokine, but also skin dryness, eczema, itching, erythema in mouse models or actual human application tests. It has an alleviating effect, and also provides a skin barrier strengthening effect and skin moisturizing effect, and does not show cytotoxicity, so it is also useful as a good treatment strategy for inflammatory skin diseases that do not cause side effects.

1 is a graph showing the inhibitory effect of NO production according to the concentration of sodium surfactin treatment in Raw 264.7 cells.
2 is a photograph showing the expression levels of IL-6, TNF-alpha, and GAPDH according to the concentration of sodium surfactin treatment in Raw 264.7 cells.
3 is a graph showing changes in body weight and ear thickness between PA untreated group, PA alone treated group, and PA and sodium surfactin treated groups by applying phthalic anhydride (PA) to the back and back of the ear of a mouse model for 4 weeks.
Figure 4 is a graph measuring the amount of IL-6 expression between PA untreated group, PA alone treated group, PA and sodium surfactin treatment group in serum at autopsy after applying phthalic anhydride (PA) to the back and back of the ear of a mouse model for 4 weeks to be.
FIG. 5 is a photograph of visual observation of skin diseases such as PA untreated group, PA alone treated group, and PA and sodium surfactin treated groups by applying phthalic anhydride (PA) to the back and back of the ear of a mouse model for 4 weeks.
6 is a photograph showing the degree of skin barrier recovery after 1 week and 2 weeks when sodium surfactin is applied after damage to the skin barrier using SLS in a human application test.
7 is a graph showing the amount of moisture increase before and after application of a cream containing sodium surfactin.
8 is a graph showing the results of measuring moisturizing over time of use of a cream containing sodium surfactin.
9 is a graph of changes in the amount of transdermal moisture loss over time of use of a cream containing sodium surfactin.

Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by an expert skilled in the art to which the present invention belongs. In general, the nomenclature used in this specification is well known and commonly used in the art.

Definitions of main terms used in the detailed description of the present invention are as follows.

The term “treatment” as used herein refers to an approach to obtain beneficial or desirable clinical outcomes. For the purposes of the present invention, beneficial or desirable clinical outcomes include, but are not limited to, alleviation of symptoms, reduction of disease range, stabilization of disease state (i.e., not exacerbation), delay or decrease in disease progression, disease state. Amelioration or temporal alleviation and alleviation of (which may be partial or total), detectable or undetectable. “Treatment” refers to both therapeutic treatment and prophylactic or preventative measures. These treatments include the disorder to be prevented, as well as the treatment required for the intestine that has already occurred. “Improving” a disease means that the extent of the disease state and/or undesirable clinical signs are reduced or the temporal transition of progression is slowed or prolonged, compared to the case without treatment.

The term "prevention" as used herein refers to a therapy that protects against the onset of a disease or disorder so that clinical symptoms of the disease do not develop. Thus, “prophylaxis” relates to administering a therapy to a subject (eg, administering a therapeutic substance) before signs of disease are detectable in the subject. A subject may be an individual at risk of developing a disease or disorder, such as an individual having one or more risk factors known to be associated with the development or onset of the disease or disorder. Thus, in certain embodiments, the term “preventing inflammatory skin disease” refers to administering an inflammatory skin disease treatment substance to a subject who does not have a detectable inflammatory skin disease. It is understood that a subject for anti-inflammatory prophylactic therapy may be an individual at risk of developing an inflammatory skin disease.

As used herein, the term “therapeutically effective amount” or “effective amount” is an amount effective to elicit a desired biological or medical response, such as sodium surfactin sufficient to produce such treatment for a disease when administered to a subject for treating the disease. Refers to sheep. The effective amount will depend on the disease and its severity and the age, weight, etc. of the subject to be treated. An effective amount can include a range of amounts. As understood in the art, an effective amount may be one or more doses, i.e., a single dose or multiple doses may be required to achieve the desired therapeutic endpoint. An effective amount may be contemplated in connection with administering one or more therapeutic agents, and a single agent may be considered to be provided in an effective amount in combination with one or more other agents in which a desired or beneficial result may or be achieved. A suitable dose of any co-administered compound may optionally be reduced due to the combinatorial action (eg, additional or synergistic effect) of the compound.

Throughout this specification, unless otherwise indicated in the context, the phrases "include" and "include" are meant to imply that the presented step or group of steps is included, but no other step or group of steps is excluded. Must understand.

Hereinafter, the present invention will be described in detail.

In one aspect, the present invention relates to a pharmaceutical composition for the prevention or treatment of inflammatory skin diseases containing sodium surfactin as an active ingredient.

In the present invention, sodium surfactin is a composition characterized in that it has a structure represented by Formula 1:

[Formula 1]

Where n=9.

In an embodiment of the present invention, it was confirmed that sodium surfactin not only has the activity of inhibiting NO production when inflammation is induced using LPS, but also decreases the expression of IL-6, an inflammatory cytokine, in a concentration-dependent manner.

In another embodiment of the present invention, in a mouse model stimulated with PA, sodium surfactin was found to alleviate changes in crust formation and thickening of the dermal layer due to PA induction, and IL-6 expression in the mouse model serum was also reduced. In addition, it was confirmed that dry skin and eczema were greatly relieved by visual inspection.

In another embodiment of the present invention, it was confirmed that erythema and itching in atopic dermatitis patients were improved.

In addition, in another embodiment of the present invention, as a result of recruiting and testing 20 subjects, it was possible to confirm the skin barrier strengthening effect and skin moisturizing effect of the cream containing sodium surfactin.

Accordingly, in another aspect, the present invention relates to a cosmetic composition for improving inflammatory skin diseases or moisturizing skin, containing sodium surfactin as an active ingredient.

In the present invention, "skin disease" means an abnormal condition or symptom occurring in the skin. In one embodiment of the present invention, the skin disease is an inflammatory skin disease. The inflammatory skin disease refers to a skin disease exhibiting symptoms such as itching and erythema due to an inflammatory reaction mediated by immune cells. At this time, the inflammatory skin disease may be allergic dermatitis, atopic dermatitis, eczema, athlete's foot, allergic urticaria, pruritus, ringworm, psoriasis, or dry skin. Here, atopic dermatitis is a skin eczema disease accompanied by severe itching that recurs chronically, and is a part of dermatitis.

The composition of the present invention can be prepared as a pharmaceutical composition.

According to a preferred embodiment of the present invention, the composition of the present invention comprises (a) a pharmaceutically effective amount of sodium surfactin of the present invention described above; And (b) is a pharmaceutical composition comprising a pharmaceutically acceptable carrier.

When the composition of the present invention is prepared as a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier included in the pharmaceutical composition of the present invention is usually used in formulation. As used, lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl Including, but not limited to, cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, etc. The pharmaceutical composition of the present invention is not limited to the above ingredients, but also lubricants, humectants, sweeteners, flavors. Agents, emulsifiers, suspending agents, preservatives and the like may further be included. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed, 1995).

The pharmaceutical composition of the present invention can be administered parenterally or orally, such as skin administration, intravenous administration, subcutaneous administration, intramuscular administration, intraperitoneal administration, topical administration, intranasal administration, intrapulmonary administration, rectal administration, dura mater It can be administered by methods such as intramuscular administration, ocular administration, and transdermal administration. In clinical practice, the sodium surfactin of the present invention can be administered by any conventional route, especially as an external agent.

The sodium surfactin is administered to a subject or patient, wherein the “subject(s)” are animals such as non-primates (eg cows, pigs, horses, cats, dogs, rats and mice) and primates (E.g., monkeys, such as cynomolgous monkeys, chimpanzees and humans), including mammals, and, for example, humans. In another embodiment, the subject is a livestock animal (eg, horse, cow, pig, etc.) or a pet animal (eg, a dog or cat). In a preferred embodiment, the subject is a human.

A suitable dosage of the pharmaceutical composition of the present invention may be prescribed in various ways depending on factors such as formulation method, administration mode, age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity. The general dosage of the pharmaceutical composition of the present invention is within the range of 00001-1000 mg/kg on an adult basis, but is not limited thereto.

The pharmaceutical composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person skilled in the art. In this case, the formulation may be in the form of a solution, suspension, syrup or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, powder, granule, tablet or capsule, and a dispersant or Stabilizing agents may additionally be included.

The composition of the present invention may be prepared as a cosmetic composition. When the composition for preventing or improving inflammatory skin diseases comprising sodium surfactin of the present invention as an active ingredient is prepared as a cosmetic composition, a cosmetic composition other than sodium surfactin as an active ingredient Ingredients commonly used in, for example, stabilizers, solubilizers, conventional adjuvants such as vitamins, pigments and flavors, and carriers.

The cosmetic composition of the present invention may be prepared in any formulation commonly prepared in the art, for example, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing , Oil, powder foundation, emulsion foundation, wax foundation, and spray may be formulated, but are not limited thereto. More specifically, flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing It can be prepared in the form of a cream, cleansing foam, cleansing water, pack, spray or powder.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as carrier components. I can.

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as a carrier component. In particular, in the case of a spray, additional chlorofluorohydrocarbon, Propellants such as propane/butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, ethoxylated isostearyl alcohol, suspending agents such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, crystallites Sex cellulose, aluminum metahydroxide, bentonite, agar or lacant, and the like may be used.

When the formulation of the present invention is a surfactant containing cleansing, as a carrier component, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters may be used.

Example

Hereinafter, the present invention will be described in more detail through examples. These examples are for illustrative purposes only, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not construed as being limited by these examples.

Confirmation of the cytotoxicity of sodium surfactin

The following experiment was conducted to confirm the cytotoxicity of sodium surfactin (Kaneka corporation).

After suspending mouse fibroblast L929 cells (mouse fibroblast L929, ATCC CCL-1, USA) in DMEM containing 10% FBS, the concentration of cells in each well of a 96-well tissue culture plate is 5×10 ⁴ cells/ After dispensing to become a well, it was incubated for 24 hours under conditions of 5% CO ₂ and 37°C. After cultivation, the medium was treated with sodium surfactin at the concentrations shown in Table 1 and cultured for 24 hours. After the culture was completed, 10 µl of MTT (2 mg/ml in PBS, Thiazolyl Blue Tetrazolium Bromide, Sigma-aldrich, USA) was added to each well and reacted for 3 hours. MTT forms formazan in response to mitochondria in completely living or intact cells. After the reaction, the culture medium was removed, and 100 µl of DMSO (Dimethyl Sulfoxide, Sigma-aldrich, USA) was added to react for 10 minutes, and the absorbance was measured at 540 nm using an ELISA reader (ELX 808, BioTek Instruments Inc., USA). The cytotoxicity values for each concentration were measured and shown in Table 1 below.

Concentration (w/%) 0.001 0.01 0.1 One MTT50 Survival rate (%) 100 100 95 72 >1

As a result, as shown in Table 1, sodium surfactin according to the present invention exhibited MTT50 at 300 µg/ml or more, and thus it was found to be non-toxic or non-toxic.

In addition, cytotoxicity was measured and compared in the same manner as described above using each biosurfactant.

Surfactants NR50(g/L, w/w) Linear Alkyl Sulfonate (LAS) <0.01 Sodium Dodecyl Sulfate (SDS) 0.05 Na-Surfactin >10 Glycerol 50

As a result, as shown in Table 2, it was also found to be non-toxic or non-toxic even when compared with conventional surfactants.

That is, it can be seen that sodium surfactin according to the present invention is a very safe raw material.

Confirmation of the effect of sodium surfactin on improving inflammatory skin diseases (1)

2-1: Measurement of the inhibitory activity of Na-surfactin on the production of Nitric oxide (NO)

The degree of inhibition of inflammation was tested using the Raw 264.7 cell line, which is a general mouse macrophage.

The amount of NO produced from Raw 264.7 cells (ATCC, TIB-71) was measured in the form of NO ² present in the cell culture solution using griess reagent according to the method of Green et al. Raw 264.7 cells were aliquoted into a 6 well plate at 1 × 10 ⁵ cells/well. After incubation for 24 hours in a 37°C CO ₂ incubator, it was washed twice with 1X phosphate buffered saline (PBS). After 2 hours of treatment with 1 μg/ml of LPS except for the control (normal), a sample solution prepared by concentration (sodium surfactin) was treated and incubated for 24 hours to obtain a supernatant, and then the same amount of griess reagent was added to 96 wells. After 10 minutes reaction on the plate, the absorbance at 540 nm was measured.

As a result, as shown in FIG. 1, when Na-surfactin was treated at a concentration of 6.25, 12.5, 25, and 50 μg/ml in Raw 264.7 cells, 22.9 μM was generated in LPS alone treatment, but 22.7 and 20.8 in these experimental groups. , 18.0, 16.1 μM was measured to confirm that the concentration-dependent decrease. That is, it can be seen that sodium surfactin according to the present invention effectively inhibits NO, indicating that sodium surfactin has an anti-inflammatory effect.

2-2: Measurement of the inhibitory activity of Na-surfactin on IL-6 expression

Additionally, the expression levels of IL-6, TNF-α, and COX-2 according to Na-surfactin treatment were confirmed using Raw 264.7 cell line.

RAW 264.7 cells were dispensed in a 100mm cell culture plate at a concentration of 5×10 ⁵ cells/well and cultured for 24 hours, and LPS (1 μg/ml) and samples were each treated for 24 hours. Samples after sample treatment are washed with PBS and then RIPA buffer [Radio Immuno Precipataion Assay, 50mM Tris/HCl (pH 7.2), 150mM NaCl, 1% Triton X-100 (or NP-40), 1% Sodium deoxy cholate, 0.1 % SDS, 1% Trasylol (protease inhibitor)] was suspended with 100 ul, transferred to a 1.5 ml eppendroff tube, and reacted at 4° C. for 1 hour. The supernatant was recovered by centrifugation at 4° C. and 13,000 rpm for 5 minutes using a high-speed centrifuge, and protein quantification was measured by diluting the ratio of the sample dilution solution and the reagent to 1:9 with Bradford reagent (Sigma).

The prepared sample was mixed in the same amount with 1×Sample buffer [1M Tris/HCl (pH 6.8), 50% glycerol, 10% SDS, 2-mercaptoet hanol, 1% bromphenol blue in 3rd DW], heated for 5 minutes, and then western 10% SDS-PAGE (80V for 30 minutes, 120V for 2 hours) was performed using electrophresis (20 ug/lane of protein, BIO-RAD). The developed protein was transferred to the PVDF membrane at 100 V for 1 hour. The membrane was blocked with 5% skim milk for 1 hour at room temperature, and 1 ^st antibody (IL-6, TNF-α, GAPDH, 1:1000, Santacruze) was treated overnight at 4°C shaker. After that, the secondary antibody was reacted at room temperature for 1 hour.

As a result, as shown in Figure 2, when treated with sodium surfactin (Na-surfactin) at a concentration of 6.25, 12.5, 25, 50 ㎍ / ㎖ in Raw 264.7 cells, IL-6 and TNF-alpha expression It was confirmed that the LPS was increased in the treatment group, but decreased in a concentration-dependent manner by treatment with Na-surfactin.

Confirmation of the effectiveness of sodium surfactin in improving inflammatory skin diseases (2)

To confirm the effect of improving inflammatory skin diseases of sodium surfactin, sodium surfactin was treated in a PA (phthalic anhydride)-treated mouse model as follows, weight change, ear thickness change, cytokine measurement in serum after autopsy, and visual inspection. Was performed.

Specified pathogen-free (SPF) hairless mice (8 weeks old, 25±5g) were purchased from Central Laboratory Animals (Seoul, Korea), and irradiated feeds (Purina, Seoungnam, Korea) were fed freely. They were reared at a 12-hour lighting cycle (08:00-20:00), a temperature of 23±1℃, and a relative humidity of 50±5%. Experimental animals were randomly assigned to 3 groups (Vehicle treatment group, phthalic anhydride (PA) treatment group, and PA+ Na-surfactin treatment group). In the vehicle treatment group, purified water was applied to the ears and head of hairless mice 12 times for 4 weeks at 100 μL/1 time, and the PA treatment group was treated with 12.5% PA solution (100 μL) under the same conditions. The PA+Na-surfactin-treated group treated 12.5% of PA while simultaneously applying the cream to the skin every other day for 4 weeks. After 4 weeks, mice were anesthetized by administering Zoletil 1 mL/kg, and blood was collected. (Ear thickness measurement: The thickness of the ear was measured once a week, using a Vernier calipers (Germany) to measure the mid-central part of the ear.)

First, as a result of applying PA for 4 weeks to the back and back of the ear of the mouse model, in the case of body weight, as shown in FIG. 3A, the untreated control group (Con), the PA alone treatment group (PA), the PA and sodium surfantine treatment group (Na- Surfactin) showed little change. However, as shown in Fig. 3B, in the case of the ear thickness, in the PA-only treatment group (PA), while the PA induction was performed, a crust was gradually formed, and fat cells were pinched in the dermal layer, showing a change in thickness, but the experimental group PA and sodium surfactin Treatment group (Na-Surfactin) showed a tendency to alleviate.

In the mouse model 4 weeks after autopsy, the serum was separated and ELISA was performed using a mouse IL-6 Quantikine ELISA kit (R&D systems, Inc. USA) to measure IL-6 levels. While the mouse was anesthetized with Zoletil, blood was collected from the abdominal vein, left for 30 minutes in a heparin tube, and then centrifuged to separate the serum. First, capture antibodies were dispensed into an immunoplate and attached for 1 hour at room temperature, and unattached antibodies were used three times with a washing solution (50 mM Tris, 0.14 M NaCl, 0.05% Tween 20, pH 8.0). It was removed by washing. Serum isolated from mice and diluted standard IL-6 were added to each well of the immunoplate conjugated with the capture antibody and bound for 1 hour, then the unattached solution was removed, and the horseradish peroxidase-bound antibody Was added. Finally, a substrate solution was added and the reaction was carried out for 30 minutes at room temperature in the dark. The reaction result was confirmed by measuring the absorbance at 450 nm using a MicroplateReader (Molecular Device Co., USA).

As a result, as shown in FIG. 4, it was confirmed that IL-6 expression was significantly reduced in the PA and sodium surfactin treatment group (Na-Surfactin) compared to the PA treatment group (PA).

In addition, PA was applied to the back of a mouse model for 4 weeks and observed. As shown in FIG. 5, skin dryness and eczema occurred severely in the PA alone treatment group (PA), but the PA and sodium surfactin treatment group (Na-Surfactin ), it was confirmed that this phenomenon was greatly alleviated.

Confirmation of the skin barrier strengthening effect of sodium surfactin

Sodium surfactin-containing cream is prepared by adding 3% concentration of sodium surfactin to a cosmetic composition of a conventional cream formulation to confirm the treatment/improvement efficacy of skin diseases such as dry skin or eczema. The barrier strengthening effect was tested. After requesting the Chungcheongbuk-do Cosmetics Clinical Research Support Center, a total of 20 subjects were recruited, and the skin barrier was damaged, and the transdermal moisture loss (TEWL) of the whole skin was measured and evaluated using a Tewameter.

All evaluations were conducted under constant temperature and humidity (22±2℃, RH40~60%) conditions. First, by visually evaluating the skin condition of the entire abdominal region and conducting a basic questionnaire, 20 subjects to participate in the test were selected by evaluating factors that may affect the test (points, wounds, presence of dermatology treatment, etc.). The whole-thin part to be tested was washed and acclimated for at least 30 minutes under constant temperature and humidity conditions. Thereafter, the TWEL of the pre-cut area was measured before the test, and a 1% SLS (Sodium- Lauryl Sulfate) solution was patched on the pre-cut site.

After removing the patch, wait for at least 1 hour under constant temperature and humidity conditions, and then measure the TWEL of the test site again. Then, after applying a cream containing 3% sodium surfactin, the presence or absence of side effects was evaluated.

The improvement rate of skin barrier strengthening was calculated according to the following equation.

Skin barrier enhancement improvement rate (%) = (TEWL measurement immediately after SLS treatment-TEWL measurement after sodium surfactin) / (TWEL measurement immediately after SLS treatment-TEWL measurement before SLS treatment) * 100

The paired t-test method was used for changes before and after use, and the statistical results were considered to be statistically significant when the significant difference of 5% (p<0.05), which is the most used in biological statistical analysis.

Measurement point Before use Immediately after SLS treatment 1 week after product use 2 weeks after product use Product use Sodium surfactin No application Sodium surfactin No application Sodium surfactin No application Sodium surfactin No application TEWL measured value (mean ± standard deviation, g/m ³ /h) 6.26±1.56 6.08±1.27 48.24±13.64 47.96±14.55 13.76±3.53 14.05±3.47 8.70±1.79 10.32±2.39 Skin barrier strengthening improvement rate (%) - - - - 82.14 80.96 94.20 89.87 p value - - - - p<0.01 p<0.01 p<0.01 p<0.01

As a result, as shown in Table 3, the skin barrier strengthening effect of 82.14% after 1 week of use and 94.20% after 2 weeks of use was exhibited. When compared with the untreated control site, the effect of improving the skin barrier strengthening of the test site was statistically significantly different after 2 weeks of use. In addition, no skin adverse events occurred during the test period. 6 is an example of the results of a human application test, and shows that when sodium surfactin of the present invention is used, the improvement effect on skin damage is clear compared to the unused area.

Therefore, sodium surfactin is considered to be a substance that helps to strengthen skin barrier from 2 weeks after use.

Confirmation of the moisturizing effect of sodium surfactin

5-1: Measurement of moisturizing power of cream formulation containing sodium surfactin

A cream containing sodium surfactin was prepared by adding sodium surfactin at a concentration of 3% to a cosmetic composition of a conventional cream formulation. Thereafter, skin moisturizing power tests were performed using Skin-O-Mat (pH, moisture, oil measurement equipment, Cosmomed, Germany).

The Skin-O-Mat device is a method of measuring skin moisture using the skin capacitance. It is used for atopy, moisturizing, etc., and the degree of skin improvement can be verified objectively and reliably. . After marking the arm at regular intervals, a certain amount of the prepared sodium surfactin-containing cream was evenly applied, and after a period of time, the moisture content of the skin was measured through an equipment.

As a result, as shown in FIG. 7, when the cream containing sodium surfactin was applied, an average moisture increase rate of 69.1% was confirmed compared to before application.

5-2: Human body application test to improve moisturizing and transdermal moisture loss of cream formulation containing sodium surfactin

In order to confirm the effectiveness of sodium surfactin in the treatment/improvement of skin diseases such as dry skin or eczema, the effects of moisturizing and improving transdermal moisture loss were tested. A total of 20 subjects were recruited by requesting the Chungcheongbuk-do Cosmetics Clinical Research Support Center to perform a test for moisturizing subjects with agenda and TWEL effectiveness.

The cream containing sodium surfactin according to the present invention was used in the morning/evening, and stored according to a general cosmetic storage method. Through a subjective questionnaire, the moisture content of the whole skin is measured using a Corneometer that objectively measures the skin moisture content of the subject, which the subject perceives as dry, and the subject with the measured value less than 30 is selected as a very dry subject. Was carried out. Skin moisture content by time was measured using a Corneometer, and the amount of transdermal water loss was evaluated by measuring the transepidermal water loss (TEWL) at the test site using a Tewameter. All tests were conducted under constant temperature and humidity (22±2℃, RH 40~60%).

The effectiveness of the cream containing sodium surfactin was evaluated by measuring and analyzing the moisture content and TEWL of the test site before and after use. Moisturizing and TEWL improvement rates were calculated according to the following equation.

Moisture improvement rate (%) = (Moisture content after product use-Moisture content before product use) / (Moisture content before product use) * 100

Skin barrier (transdermal moisture loss TEWL) improvement rate (%) = (-) (TEWL after product use-TEWL before product use) / (TEWL before product use) * 100

The paired t-test method was used for changes before and after use, and the statistical results were considered to be statistically significant when the significant difference of 5% (p<0.05), which is the most used in biological statistical analysis.

Measurement point Before use 1 week after use 2 weeks after use Moisture content measurement value (average ± standard deviation. Unit: A.U.) 27.32±2.65 40.22±5.92 43.44±5.35 Skin moisture improvement rate (%) - 47.37 59.14 p value - p<0.01 p<0.01

As a result, as shown in Table 4 and FIG. 8, the skin moisture content was improved by 47.37% after 1 week after use and 59.14% after 2 weeks after use. In statistical analysis, the moisture content increased statistically significantly from 1 week after use compared to before use. During the evaluation period, no skin adverse events occurred for this product.

Measurement point Before use 1 week after use 2 weeks after use Moisture content measurement value (average ± standard deviation. Unit: A.U.) 8.97±1.25 8.16±1.39 6.72±1.87 Skin moisture improvement rate (%) - 9.08 25.13 p value - p<0.01 p<0.01

In addition, as a result of evaluating the skin barrier improvement effect by measuring the amount of transdermal moisture loss by sodium surfactin, as shown in Table 5 and FIG. 9, in terms of the amount of transdermal water loss both after 1 week of sodium surfactin and after 2 weeks of use, Showed improvement effect. In addition, from 1 week after use, the moisture content was increased statistically significantly compared to before use. In addition, no adverse skin events occurred during the evaluation period.

Confirmation of the effect of sodium surfactin on erythema and itching in patients with atopic dermatitis

The sodium surfactin-containing cream of Example 5 was used to confirm the effect of improving erythema and itching in men in their 50s suffering from atopic dermatitis. The patient complained of symptoms such as itching, erythema, scratching on the inflamed area, and wound on the scratched area with a medical history of 12 years.

The cream containing sodium surfactin was applied to the patient on the back of the left calf where atopic dermatitis occurred, 3 times a day, for 4 days, and then the improvement of erythema was visually confirmed, and the improvement of itching was confirmed by SCORE.

As a result, after applying the cream containing sodium surfactin for 4 days, it was confirmed visually that the erythema of the applied area was improved, and the itching was also confirmed to be resolved over time.

As described above, a specific part of the content of the present invention has been described in detail. For those of ordinary skill in the art, this specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereby It will be obvious. Accordingly, it will be said that the substantial scope of the present invention is defined by the appended claims and their equivalents.

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Claims (5)

A pharmaceutical composition for the prevention or treatment of atopic dermatitis through skin moisturization containing sodium surfactin as an active ingredient. delete The composition of claim 1, wherein the sodium surfactin has a structure represented by Chemical Formula 1:
[Formula 1]

Where n=9. A cosmetic composition for improving atopic dermatitis by moisturizing the skin containing sodium surfactin as an active ingredient.
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