KR20050110149A - A cosmetic composition containing an extract of sorbus commixta - Google Patents

Abstract

The present invention relates to a rowan extract and a cosmetic composition containing the same as an active ingredient, more specifically, a deciduous broad-leafed arborescent of Rosaceae as an active ingredient, the extract of the stem and bark of the rowan 0.001 to 30.0% by weight, It relates to a cosmetic composition excellent in antioxidant, anti-wrinkle effect, whitening effect, hair growth effect, acne prevention effect, stimulating alleviation, characterized in that it contains 0.01 to 5% by weight.

According to the present invention, rowan extract has anti-oxidant activity as well as human fibroblast activity effect, skin fine wrinkle improvement effect, whitening effect, hair growth effect, acne prevention effect, skin irritation relieving effect can be used to prepare various functional cosmetics using this substance have.

Description

Cosmetic composition containing rowan extract {A COSMETIC COMPOSITION CONTAINING AN EXTRACT OF SORBUS COMMIXTA}

The present invention relates to a cosmetic composition containing the plant extract as the main active ingredient.

Human skin has various physical and chemical changes in the aging process, and its causes are classified into internal aging and photo-aging. That is, it may be caused by ultraviolet rays, stress, disease state, environmental factors, wounds, activation of free radicals with age, etc., and when this condition is intensified, it destroys the antioxidant defense network existing in the living body and destroys cells and tissues. Damage to promote adult disease and aging. More specifically, lipids, proteins, polysaccharides, and nucleic acids, which are major constituents of the skin, are oxidized to destroy skin cells and tissues, resulting in skin aging. In particular, the oxidation of protein is caused by the breakdown of collagen, hyaluronic acid, elastin, proteoglycan, and fibronectin, which are the tissues of the skin, causing severe excessive inflammatory reactions and disturbance of skin elasticity. This leads to a situation of reduced immune function.

Therefore, cell membranes are protected by eliminating free radicals generated by metabolic processes in the body, ultraviolet rays, and inflammatory reactions.Also, damaged cells must be regenerated by active metabolism to proliferate the cells. You can recover quickly and maintain healthy skin.

On the other hand, aging involves not only free radicals, but also an enzyme called MMP. In vivo, the synthesis and degradation of extracellular substrates such as collagen is properly regulated, but the synthesis decreases as aging progresses, and the substrate metal protein, an enzyme that degrades collagen. Expression of the degrading enzyme (MMP) is promoted, which reduces the elasticity of the skin and forms wrinkles. In addition, these degrading enzymes are also activated by ultraviolet irradiation. Therefore, there is a need for the development of a substance capable of regulating MMP expression that inhibits activation or inhibiting its activity in cells.

Next, skin aging due to pigmentation may be mentioned, and pigments affecting skin color include melanin, melazoid, carotene, hemoglobin, and the like. The most important thing is melanin, the biggest factor affecting biosynthesis is ultraviolet radiation and hormone secretion in the body. Melanin plays an important role in absorbing or scattering ultraviolet rays to prevent damage to the skin from ultraviolet rays. There is no special maximum absorption wavelength and it absorbs light in all areas. In addition, it is excellent in removing the active oxygen species, but sometimes melanin itself generates free radicals, and other substances are reduced or oxidized by catechol or quinone in the melanin structure, and melanin itself shows the properties of free radicals. Pray.

The biosynthesis of melanin is a series of oxidation processes that begins with the conversion of tyrosine, an amino acid, from the melanocytes of melanocytes to tyrocinase and to dihydroxy phenylalanine, followed by brown (pheomelanin) and black (eumelanin). It is formed of a polymer of). This biosynthesis process is carried out in a special type of brown intracellular organelles called melanosomes. Melanosomes, including melanin granules, move from the periphery of the nucleus to the dendritic end, and then into the cytoplasm by the phagocytosis of keratinocytes. Accumulate around the nucleus of the site.

Ascorbic acid (JP-A 4-9320), hydroquinone (JP-A 6-192062), kojic acid (JP-A-56-7710), arbutin (JP-A 4-93150), and some extracts are already tyrosinase. It is used as a whitening cosmetic because it has an inhibitory activity, but its stability in cosmetic formulations is poorly degraded and colored, or its use is limited due to the occurrence of off-flavor, efficacy at the biological level, unclear effect and safety problems.

In addition, male alopecia is male hormone-dependent, and thus has a direct relationship with the amount of male hormone. Accordingly, many studies have been reported for the prevention and treatment of hair loss through the inhibition of male hormone activity. On the other hand, when the function of the sebaceous gland is increased by the increase in the secretion of male hormones, the scalp produced by stagnation in the hair follicles due to the hyperkeratosis of the hair follicle wall is an early stage of acne. To explain the mechanism of hair loss and acne caused by male hormones, 5alpha-Reductase is present in testosterone-reactive tissues such as sebaceous glands, hair follicles, prostate, and epididymis, and is a testosterone ( is an enzyme involved in metabolizing testosterone into dihydrotestosterone, and its conversion requires NADPH. In addition, testosterone is involved in male sexual impulses, skeletal muscle increase, male external genitalia, scrotum growth, spermatogenesis, etc., and dihydrotestosterone (dihydrotestosterone) is involved in such tissues as acne, sebum increase, hair loss and prostatic hypertrophy ( Diane et al. JID 1995). Therefore, studies are being actively conducted to develop sebum secretion inhibitors and anti-hair loss agents using inhibitors of this enzyme. Excessive secretion of male hormones after puberty leads to acne and hair loss, using a 5-alpha-reductase inhibitor to prevent excessive production of dihydrotestosterone, the active form of male hormones, by 5 alpha-reductase. And researches to develop acne preventatives are active.

Until now, most of the raw materials used as materials for cosmetics simply inhibited enzyme activity, and there was a problem in that the amount of expression of enzymes could not be controlled. In addition, consumers are increasingly demanding the use of cosmetics with minimal functional side effects such as skin protection, irritation suppression, inflammation suppression, and sun protection while having high quality functional properties according to special composition, and to minimize skin irritation As consumers' response to cosmetics using natural materials instead of using chemical raw materials is increasing, there is a need to develop natural materials as raw materials for cosmetics.

 The present invention has been made in order to solve the above problems, the object of the present invention is to select the extract prepared from the natural product of the Rowan deciduous leaf chlorophyll of the Rowan family of several natural products, to confirm the efficacy as a raw material of cosmetics The present invention provides a method of developing a method of using the same and producing a cosmetic having a predetermined functionality by using this raw material.

That is, an object of the present invention is to provide a natural extract that is applicable to skin cosmetics and exhibits excellent anti-aging effects such as antioxidant effects, collagen synthesis effects, skin fine wrinkles effects when contained in cosmetics.

In addition, it is an object of the present invention to provide a natural extract that exhibits a whitening effect by ultraviolet irradiation and the like to alleviate skin irritation by an inflammation mediator when applied to cosmetics.

It is another object of the present invention to provide a natural extract that exhibits 5 alpha-reductase inhibitory effect with excellent hair loss prevention and acne prevention effect.

Cosmetic composition according to the present invention for achieving the above object is characterized by containing the rowan extract as the main active ingredient.

Recently, many cosmetic products using natural products have been developed to reduce skin irritation caused by various chemicals. Natural ingredients have less side effects on the skin, and as the consumer's response to cosmetics using natural ingredients has increased recently, the development value of cosmetics as a raw material for cosmetics is increasing. Therefore, the present inventors have studied the possibility of applying to cosmetics in a variety of natural products that have not been known so far, as a deciduous broad-leafed arborescent of Rosaceae, select the rowan natural products of stems and bark, and extracts from them, the skin Antioxidant effect, collagen synthesis effect, skin fine wrinkle relieving effect, whitening effect, hair growth effect, acne prevention effect and skin irritation relieving effect were measured and found that it can be expected to be effective as a cosmetic.

Rowan is a deciduous broad-leaved arborescent of the Rosaceae family and its scientific name is SORBUS COMMIXTA. It is a deciduous arborescent growing up to 10m in height, but it grows in shrubs of 2-3m in high altitude, with little hairs on young branches, and two years are dark brown, and the trunk is grey, and there is a big tree. The leaves are 12 ~ 24cm in length of right upper lobe, 5 ~ 7 pairs of leaflets are the largest in the middle, and the length of the leaflets is 2.5 ~ 8cm long, ellipsoidal, with sharp tip and bottom, without leaf disease, with fusiform and hairless on both sides. Flowers hang on the end of branch in 8 ~ 12cm diameter Boksan Fire Station, without hairs, without diameter, 6 ~ 10mm, white, short flower. Calyx is cup-like, divided into 5, lobe is broad triangle, 5 petals are oblate, and hair is inside. It has 20 stamens, 3 ~ 4 shippers, hairs on the base, and the fruit is spherical and 5 ~ 6mm in diameter. The bark of the rowan in the oriental medicine is called affia is used for bronchitis, rheumatoid arthritis, stroke, gastritis, bosin, bone pain, fruit can be used as tea or reproduction.

The mountain ash extract used in the present invention is a leachate obtained by leaching and transferring from rowan, or a concentrate obtained by partially or totally concentrating the leachate, or an extract extracted and dried by extracting the concentrate. It contains the chemical itself that works.

On the other hand, the content of the mountain ash extract according to the present invention is preferably 0.001 to 30.0% by weight based on the total freeze-drying weight of the cosmetic composition. When the content of the extract is less than 0.001% by weight, there is almost no skin improvement effect, and when the content of the extract is more than 30.0% by weight, it is not economical because the degree of increase of the effect on the content is minimal.

In addition, the cosmetic composition according to the present invention may contain, in addition to the above-mentioned extract, other components that can give a synergistic effect to the main effect, preferably within the range of not impairing the main effect of the present invention.

The rowan extract may be present in the composition at 0.001 to 30.0% by weight based on the total lyophilized weight. If the plant extract is present in the form of a solution containing the extract, those skilled in the art will be able to adjust the amount of this solution in the composition according to the invention within the concentration range.

In addition, the extract of the mountain ash according to the present invention extracted with dried 10-95% (V / V) ethanol aqueous solution from 70 to 85 ℃ and cooled, the resulting filtrate was concentrated under reduced pressure below 50 ℃ and filtered under reduced pressure and reduced pressure The concentrate is prepared by lyophilization.

The cosmetic composition of the present invention uses a solvent selected from purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane, hexane and mixtures thereof.

The subject of the present invention is also the use of the cosmetic composition used as a skin lightening agent, anti-hair loss, anti-aging agent, skin irritant.

The compositions of the present invention may be in the form of an aqueous, aqueous-alcoholic or oily solution, oil-in-water or oily water or multiple emulsions, aqueous or oily gels, liquid, pasty or solid anhydrous products, oil dispersions in aqueous phase using globules. And more preferably in the form of lipid vesicles in ionic and / or nonionic form.

The composition may be somewhat fluid and may have the appearance of a white or colored cream, ointment, milk lotion, serum, essence, paste or mousse. It may optionally be applied to the skin in the form of an aerosol or may be in the form of a solid, such as a stick. It may be used as a skin care product and / or makeup product.

In a known manner, the compositions according to the invention are also useful in conventional cosmetics such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic active agents, preservatives, antioxidants, solvents, fragrances, fillers, blockers, pigments, odorants and dyes. It may contain. The amounts of these various adjuvants are amounts conventionally used in the art, such as from 0.01% to 20% by weight relative to the total weight of the composition. Depending on their nature, such adjuvants may be incorporated into the fatty phase, aqueous phase, lipid vesicles and / or nanoparticles. The adjuvant and its proportions in any case may be selected so as not to adversely affect the desirable properties of the cosmetic composition according to the invention. will be.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these Examples are only illustrative description of the present invention and the scope of the present invention is not limited to these Examples.

Example 1: Preparation of rowan extract

The dried and dried rowan were refluxed three times for 5 hours with 95% (V / V) ethanol aqueous solution, and then cooled and filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure and freeze dried at 50 ° C or lower. The ash extract was prepared by dispersing / dissolving the depressurized concentrate and the lyophilized product by using at least one solvent selected from purified water, ethanol, butylene glycol, and propylene glycol.

Example 2 Experiment of Measuring Antioxidant Effect Using NBT Method

In this example, to measure the antioxidant effect of the rowan extract obtained in Example 1, the antioxidant activity such as retinol and BHT in a comparative sample was compared, and the antioxidant activity was measured using the NBT method.

In order to measure the antioxidant effect, the active oxygen produced by xanthine and xanthine oxidase is measured by the NBT method, and the effect of the test substance removing the active oxygen, that is, the active oxygen scavenging effect, is evaluated. Free radicals are generated by xanthine and xanthine oxidase, and the active oxygen scavenging rate is measured by measuring the blue color produced by reaction of the reactive oxygen with Nitro Blue Tetrazolium (NBT) at a wavelength of 560 nm. .

As a measuring method,

0.05M Na ₂ CO ₃ ------------------------------------ 2.4ml

 2.3mM xanthine solution ---------------------------------- 0.1ml

 3.3mM EDTA Solution ------------------------------------ 0.1ml

 4.BSA solution ---------------------------------------- 0.1ml

 5.0.72 mM NBT solution ---------------------------------- 0.1ml

 6.Xanthine Oxidase Solution ----------------------------- 0.1ml

7.6 mM CuCl ₂ solution ----------------------------------- 0.1ml

① Add 1,2,3,4,5 to Bayer's bottle and add 0.1ml of sample solution to it and leave at 25 ℃ for 10 minutes.

② Add 6 and stir quickly, and start incubation for 20 minutes at 25 ℃.

③ After that, add 7 to stop the reaction and measure the absorbance St at 560nm.

④ For the blank test, measure the absorbance Bt by using distilled water instead of the sample solution as above.

⑤ Also, the blank of the sample solution is operated in the same manner using distilled water instead of 6 to measure the absorbance Bo.

The result of the effect was calculated by Equation 1, the results are shown in Table 1.

% Inhibition = [1- (St-So) / (Bt-Bo)] X 100

St: absorbance at 560 nm after enzyme reaction of sample solution

Bt: absorbance at 560 nm after enzymatic reaction of blank test solution

So: absorbance at 560 nm before reaction without enzyme in sample solution

Bo: absorbance at 560 nm before reaction without enzyme in blank test solution

As a result, as shown in Table 1, all of the samples tested at 0.1% treatment concentration showed excellent antioxidant effect, and rowan extract was confirmed to have an excellent antioxidant effect similar to retinol and BHT.

       Sample Name Treatment Concentration (%) Antioxidative effect(%)      Rowan Extract (Example 1)      0.1       93        Retinol      0.1       93         BHT      0.1       90

Example 3: Antioxidant Effect Measurement Experiment Using DPPH Method

In this example, antioxidant activity of green tea extract and antioxidants such as vitamin E was measured using a DPPH method in order to measure the antioxidant effect of the rowan extract obtained in Example 1.

DPPH method measures the antioxidant activity by reducing power using a free group called 2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl free radical (DPPH). The free radical scavenging rate is measured at a wavelength of 560 nm by comparing the degree to which the DPPH is reduced by the test substance to reduce the absorbance.

As a reagent used, a 0.1 mM solution of 2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl free radical (Aldrich Chem. Co., MW = 618.76) was dissolved in methanol to make 100 ml.

As a measuring method

① Add 0.15ml of sample solution to 0.15ml of 0.1mM DPPH solution in 96-well plate, stir rapidly, and incubate for 10 minutes at 25 ℃.

(2) Then measure the absorbance St at 560nm.

③ In the blank test, measure the absorbance Bt by using distilled water instead of the sample solution as above.

④ Again, the blank of the sample solution was operated in the same manner using methanol instead of the 0.1 mM DPPH solution to measure the absorbance Bo.

The result of the effect was calculated by Equation 2, the results are shown in Table 2.

% Inhibition = [1- (St-So) / (Bt-Bo)] X 100

St: absorbance at 560 nm after free radical scavenging of sample solution

Bt: absorbance at 560 nm after free radical scavenging of blank test solution

So: Absorbance at 560 nm before reaction without free radicals in sample solution

Bo: absorbance at 560 nm before reaction without free radicals in blank test solution

As shown in Table 2, rowan extract had better antioxidant effect than green tea extract and vitamin E at 0.01% concentration.

Sample Name Treatment Concentration (%) Antioxidative effect(%) Rowan Extract (Example 1) 0.01 82 Green Tea Extract 0.01 20 Vitamin E 0.01 23

Example 4: in vitro MMP-1 inhibition assay

Substrate metalloproteinase (MMP-1) inhibitory activity was determined in a biochemical model based on the use of collagen and gelatin conjugated to purified collagenase and its substrate, fluorescein (EnzChek ™ gela) Tinase / collagenase kit, molculaprobe). Collagenase purified from Clostridium histiocom was supplied in the EnzChek ™ gelatinase / collagenase kit. A reaction buffer consisting of DQ-collagen purified from porcine skin and conjugated to fluorescein with 0.05M Tris-HCl, 0.15M NaCl, 5mM CaCl ₂ and 0.2mM sodium azide (pH 7.6) was found in EnzChek® gelatinase. Collagenase kit (Molcula Probes) was used. Rowan extract was dissolved in the reaction buffer. It is 4; 2; 0.4; 0.2; Test at 0.1% (w / v). Dilutions of the test extracts were incubated with 25 ug / ml of DQ-collagen and 0.1 U / ml of collagenase for 15 minutes, 45 minutes, and 120 minutes at room temperature.

In each experimental condition, the control corresponding to the collagenase and DQ-collagen mixture was also incubated in the same manner. Also in each experimental condition a sample called Blank, hereinafter 'enzyme free blank', was incubated in the presence of DQ-collagen and in the absence of collagenase. Each experiment was performed three times.

After 15, 45 and 120 minutes, the signal corresponding to the degradation of DQ-collagen was measured with a fluorometer (excitation: 485 nm, emission 505 nm). The fluorescence value of each sample was measured based on the 'enzyme free blank' fluorescence value. Results are expressed as percent variance for fluorescence units per sample and control.

In conclusion, rowan extracts tested at 0.04 to 4% (v / v) under selected experimental conditions retained anticollagenase / gelatinase activity in a dose dependent manner.

As shown in Table 3, rowan extract inhibited 80% of collagen degrading activity of Clostridium collagenase when treated with 0.04%, which was superior to the inhibitory effect of green tea extract.

Experimental Sample % Inhibition Treatment concentration (%) 0.02 Treatment concentration (%) 0.04  Rowan Extract (Example 1) 55 80 Retinol 0 5 Green Tea Extract 52 56

Example 5: Evaluation of MMP-1 Expression Inhibition by Rowan Extract after Ultraviolet Irradiation

This example was subjected to ELISA to measure the concentration of MMP-1 after UV irradiation and sample addition of the rowan extract obtained in Example 1.

Human dermal fibroblasts are irradiated with UVA at an energy of 5 J / cm 2 using a UV chamber. UV irradiation dose and culture time were established through preliminary experiments to maximize the expression of MMP in fibroblasts. Negative controls were wrapped in tinfoil and maintained at the same time in the environment of UVA. UVA emission was measured using a UV radiometer. The cells during the UVA irradiation are intact in the previously dispensed medium and irradiated with UVA and then exchanged with the medium containing the sample for 24 hours of incubation, and then the medium is recovered and coated in 96-well.

The primary antibody (MMP-1 (Ab-5) monoclonal antibody and MMP-2 (Ab-3) monoclonal antibody) is treated and reacted at 37 ° C. for 60 minutes. After reacting the secondary antibody anti-mouse IgG (whole mouse, alkaline phosphatase conjugated) for about 60 minutes, the alkaline phosphatase substrate solution (1mg / mlρ-nitrophenyl phosphate in diethanolamine buffer solution) was reacted at room temperature for 30 minutes and then the microplate reader was reacted. Absorbance is measured at 405 nm. As a control, the one without addition of the sample is used.

As shown in Table 4, for MMP-1, which is induced during UV irradiation, rowan extract showed more than 20% inhibition rate compared to the control group, which was superior to the inhibition rate of retinol used as a control group. The result is.

Test group Treatment concentration (%) MMP-1 expression inhibition rate (%) Control - - Rowan Extract (Example 1) 0.1 25 Retinol 0.1 18

Example 6: Experiment to measure melanin production inhibitory effect using B16F1 melanocytes

This Example is to determine the whitening effect by looking at the degree of melanin inhibition on B16F1 melanocytes in order to confirm the whitening effect of the rowan extract obtained in Example 1.

B16F1 melanocytes used in this example are cell strains derived from mice, and cells that secrete a black pigment called melanin. Samples were treated during the artificial culture of these cells to compare the degree of melanin black pigment reduction. The B16F1 melanocytes used in this example were used after being sold from ATCC (American Type Culture Collection, Accession No .: 6323).

The melanin biosynthesis inhibitory effect of B16F1 melanocytes was measured as follows. B16F1 melanocytes were dispensed in 6-well plates at a concentration of 2 x 10 ⁶ per well, cells were attached, and the samples were incubated for 72 hours by treating the samples at a concentration that does not cause toxicity. After 72 hours of incubation, the cells were detached with trypsin-EDTA, the cell number was measured, and the cells were recovered by centrifugation.

Quantification of intracellular melanin was carried out with a slight modification of the method of Rotan: Cancer Res. , 40: 3345-3350, 1980. The cell pellet was washed once with PBS, and then 1 ml of homogenization buffer solution (50 mM sodium phosphate, pH 6.8, 1% Triton X-100, 2 mM PMSF) was added to vortex for 5 minutes to disrupt the cells. Melt the extracted melanin by adding 1N NaOH (10% DMSO) to the cell filtrate obtained by centrifugation (3,000 rpm, 10 minutes), and measure the absorbance of melanin at 405 nm with a microplate reader, and then quantify the melanin. Melanin production inhibition rate (%) was measured. Melanin inhibition rate (%) of B16F1 melanocytes was calculated by the equation (3), IC ₅₀ value is the concentration of the substance inhibits 50% melanin production.

% Inhibition = [(A-B) / A] x 100

A: Melanin amount in wells without sample

B: Melanin amount in the wells to which the sample was added

As a result of testing the melanin production inhibitory effect of B16F1 melanocytes, as shown in Table 5, the IC ₅₀ value of the rowan extract is 0.08%, similar or superior to the conventional whitening agents hydroquinone, arbutin, oil-soluble licorice extract, and extract of lettuce extract It can be seen that indicates.

Sample Name Treatment concentration (%) Melanin production inhibitory effect (IC ₅₀ ) Rowan Extract (Example 1) 0.1 0.08% Hydroquinone 0.1 0.03% Arbutin 0.1 0.2% Lettuce extract 0.1 5% Soluble Licorice Extract 0.1 0.03%

Example 7 Cytotoxic Alleviation Effect by UV Irradiation

This example was carried out to evaluate the mitigating effect of cytotoxicity by ultraviolet irradiation of the rowan extract obtained in Example 1. Fibroblasts were placed in 24-well test plates 1 × 10 5 and allowed to attach for 24 hours. Each well was washed once with PBS and 500 ul of PBS was added to each well. The fibroblasts were irradiated with 10 mJ / cm 2 of UV light using an ultraviolet B (UVB) lamp (Model: F15T8, UV B 15W, Sankyo Dennki, Japan), after which PBS was removed and 10% FBS was added to the cell culture medium (DMEM). 1 ml) was added. After treating the mountain ash extract to be evaluated here, it was incubated for 24 hours. After 24 hours, remove the badge, 500 μl of cell culture medium and 60 μl of MTT solution (2.5 mg / ml) were added to each well, followed by incubation in a 37 ° C. CO ₂ incubator for 2 hours. The medium was removed and 500 μl of isopropanol-HCl (0.04 N) was added. After shaking for 5 minutes, the cells were lysed and 100 μl of the supernatant was transferred to a 96-well test plate, followed by measurement of 565 nm absorbance in a microplate reader. Cell survival rate (%) was measured by the equation (4) and cytotoxicity relaxation by ultraviolet light was calculated by the equation (5).

Cell survival rate (%) = ((St-Bo) / (Bt-Bo)] X 100

Bo: 565 nm absorbance of wells that only reacted with cell culture media

Bt: 565 nm absorbance of the wells that developed the colorless reaction wells

St: 565 nm absorbance of the wells that reacted the sample treated wells

Reduction rate of cytotoxicity by ultraviolet ray (%) = [1- (St-Bo) / (Bt-Bo)] X 100

Bo: cell viability of wells without UV irradiation and no sample treatment

Bt: cell viability of wells that were not irradiated with UV light

St: Cell survival rate of wells treated with UV

Experimental results As shown in Table 6, rowan extract can be found to effectively protect against cytotoxicity caused by UV at 0.013% concentration and 55% cytotoxicity at low concentration.

Sample Name Treatment concentration (%) Cytotoxic alleviation rate (%) Rowan Extract (Example 1) 0.013 55 0.025 75

Example 8 Inhibitory Effect of Inflammatory Cytokine Expression by Ultraviolet Irradiation

This Example is to evaluate the degree of inhibition of inflammatory cytokine expression expressed by ultraviolet irradiation in order to confirm the anti-inflammatory effect of the rowan extract obtained in Example 1.

Fibroblasts isolated from human epidermal tissue (Fibroblast) were placed in a 24-well test plate each 5X10 ⁴ and attached for 24 hours. Each well was washed once with PBS and 500 μl of PBS was added to each well. The fibroblasts were irradiated with 10 mJ / cm 2 of ultraviolet rays using an ultraviolet B (UVB) lamp (Model: F15T8, UV B 15W, Sankyo Dennki, Japan), after which PBS was removed and cell culture medium (FMEM was not added to DMEM). Medium) 350 μl was added. After treating the rowan extract to be evaluated here, it was incubated for 5 hours. 150 µl of the culture supernatant was taken to quantify IL-1α to determine the inhibitory effect of inflammatory cytokine expression of rowan extract. The amount of IL-1α was quantified using Enzyme-linked Immunosorbent Assay, and the production rate of IL-1α was calculated by Equation 6.

Inhibitory rate of inflammatory cytokine expression (%) = [1- (St-Bo) / (Bt-Bo)] X 100

Bo: amount of IL-1α produced in wells not irradiated with UV

Bt: amount of IL-1α produced in wells that were not irradiated with UV light

St: Production of IL-1α in Wells Treated by Ultraviolet Irradiation

As shown in Table 7, the rowan extract inhibits 58% of the inflammatory cytokine-induced inflammatory cytokine IL-1α at a concentration of 0.013% at 58%, effectively preventing the occurrence of inflammation caused by UV at low concentrations.

Sample Name Treatment concentration (%) Inhibitory rate of inflammatory cytokine expression (%) Rowan Extract (Example 1) 0.013 58 0.025 80

Example 9: Paper Disc Test

This Example was a paper disc test (Paper Disc Test) to confirm the antimicrobial activity of acanthus of the rowan extract obtained in Example 1.

First, in order to activate propionibacterium acnes, a skin acne causing cause of acne, the cultures were pre-cultured for 48 hours in BHI liquid medium (Brain-Heart Infusion Broth; 3.7%). The culture medium thus prepared was plated in 0.1 ml each of BHI solid medium (Brain-Heart Infusion Broth; 3.7%; agar 1.5%) and dried. The ash extract obtained in Example 1 was diluted to 12% (W / v) in a 95% ethanol aqueous solution, and 50 µl each was added to a 8 mm diameter paper disc and placed on a solid medium prepared above. The cells were incubated for 3 days.

The antimicrobial activity was evaluated by observing the growth inhibition zones around the paper disc and measuring the size of the ring. As a result, the size of the bacterial growth inhibitory ring of rowan extract was found to be 22 mm.

Example 10: Evaluation of inhibitory activity of 5-alpha-reductase activity of rowan extract

This example is to evaluate the degree of inhibition of 5 alpha-reductase activity to determine the hair loss prevention and acne prevention effect of the rowan extract obtained in Example 1.

5 alpha-reductase activity inhibition 5 alpha-reductase used in the experiment was used an enzyme produced by the epidermal fibroblasts.

Fibroblasts are inoculated so that 10000 cells per microplate hole is incubated. Each hole is cultured after adding 0.1.mμ.Ci of radiolabeled testosterone with ³ H (tritium) to determine whether fibroblasts use it. As a control, do not contain rowan extract. After 24 hours of incubation, the supernatant is obtained, and steroids are obtained with 1 ml of ethylacetate-cyclohexane (1: 1) extractant. The obtained steroid is placed on a thin layer chromatography plate and developed with a chloroform / methanol mixture (98/2 (v / v)). Calculate the conversion rate by measuring the radioactivity of the testosterone and the dihydrotestosterone point using a densitometer, and compares the result with the control (conversion rate when no extract is added). Alpha-reductase inhibitory activity was evaluated.

% Inhibition = [A-B] / [A] × 100

A = conversion of testosterone to dihydrotestosterone (without extract)

B = conversion of testosterone to dihydrotestosterone (extract added)

As shown in Table 8 it can be seen that the rowan extract has a 5 alpha-reductase inhibitory effect.

Rowan extract concentration (%) One 0.1 0.05 0.01 0.005 0.001 0.0001 % Inhibition 100.0 100.0 95.0 82.5 73.8 52.6 41.5

Example 11: Hair Growth Effect Test

This example prepared a 30% hydrous ethanol solution containing 2% of the dried extract of rowan extract obtained in Example 1 was used for the test. In the hair growth effect test, 47-53 days after the birth of the mouse (ICR) were used to remove the back hair, and the back area was selected to be clean and 10 rats were used for each material group. It was applied thickly. The length of hair and the degree of hair growth over time were compared by adding scores according to the degree of restoration after hair removal. In order to compare the degree of hair growth, as a control, a 30% alcohol solution was applied to each individual to observe the growth state.

As a result, it can be seen that the rowan extract as shown in Table 9 has an excellent hair growth promoting effect.

Elapsed Days / Sample 5 10 15 Rowan extract 0.25 0.94 1.88 ± 0.44 Control 0.08 0.26 1.01 ± 0.23

Example 12 and Comparative Example 1: skin elasticity improvement effect

In this example, the cosmetics containing the rowan extract obtained in Example 1 were prepared to evaluate skin elasticity improvement effect in humans by performing a comparative experiment with Comparative Example 1.

The cosmetics used in the comparative experiments are in the form of a cream, the composition of which is shown in Table 10. First, the phase b) recorded in Table 10 is heated and stored at 70 ° C. To this, a) phase is added, and after pre-emulsification, it is emulsified uniformly with a homomixer, and then cooled slowly to prepare a cream (Example 12, Comparative Example 1). 20 experimenters (20-35 year old female) were applied to the cream prepared in Example 12 on the right side of the face and the cream prepared in Comparative Example 1 on the left side of the face, twice a day for 3 consecutive months.

The skin elasticity improvement effect after the completion of the experiment was measured using a skin elasticity measuring instrument (cutometer SEM 575, C + K Electronic Co., Germany) before and after using the product for 2 months. The experimental results are described in Table 11 as ΔR8 value of Cutometer SEM 575, where R8 value represents the nature of the viscoelasticity of the skin.

As shown in Table 11, it can be seen that the skin elasticity improvement effect of the experimenter who applied the cream containing rowan extract was excellent.

Raw material   Example 12    Comparative Example 1      end Stearyl alcohol 8 8 Stearic acid 2 2 Stearic cholesterol 2 2 Squalane 4 4 2-octyldodecyl alcohol 6 6 Polyoxyethylene (25 mol addition) alcohol ester 3 3 Glyceryl monostearic acid ester 2 2      I Rowan extract One - Propylene glycol 5 5 Quantity Quantity Quantity

      Note) Unit: Weight%

Experimental product Skin elasticity effect (ΔR8)     Example 12 0.24     Comparative Example 1 0.12

       n = 20, p <0.05

Example 13 and Comparative Example 2

In this example, the cosmetics containing the rowan extract obtained in Example 1 were prepared, and the skin whitening effect of humans was evaluated by comparative experiments with Comparative Example 2.

The cosmetics used in the comparative experiments are in the form of a cream, the composition of which is shown in Table 12. First, the phase b) recorded in Table 12 is heated and stored at 70 ° C. To this, a) phase is added, and after pre-emulsification, it is emulsified uniformly with a homomixer, and then cooled slowly to prepare a cream (Example 13, Comparative Example 2). 20 experimenters (women aged 20-35 years) were applied with the cream prepared in Example 13 on the right side of the face and the cream prepared in Comparative Example 2 on the left side of the face twice a day for 2 consecutive months. . After completion of the test, the color of the applied areas on both sides of the face was measured using an image analyzer and the color of the skin using chromameter (Minolta CR300) to measure the change in brightness of the color (ΔL). Subjective visual observation was performed and the effect was measured according to the following classification. The results are shown in Table 13 below. At this time, the degree of whitening efficacy was classified into the following seven ratings.

Evaluation criteria for whitening efficacy

-3: very worse -2: worse -1: slightly worse 0: no change

1: slightly improved 2: improved 3: highly improved

As shown in Table 13, it can be seen that the whitening effect is excellent in the facial skin of the experimenter applying the cream containing rowan extract.

Raw material Example 14 Comparative Example 2    end Stearyl alcohol 8 8 Stearic acid 2 2 Stearic cholesterol 2 2 Squalane 4 4 2-octyldodecyl alcohol 6 6 Polyoxyethylene (25 mol addition) alcohol ester 3 3 Glyceryl monostearic acid ester 2 2    I Rowan extract One - Propylene glycol 5 5 Quantity Quantity Quantity

Note) Unit: Weight%

Change in brightness of skin color (ΔL) Objective evaluation of the skilled person Subjective Evaluation of Subject Example 13 Comparative Example 2 Example 13 Comparative Example 2 Example 13 Comparative Example 2 medium 5.03 2.03 2.7 1.9 2.5 1.7

Other examples are shown below.

That is, the lotion, milky lotion and beauty liquid containing the rowan extract obtained in Example 1 were prepared in Examples 14 to 16. The lotion, milky lotion and essence containing these extracts showed excellent effects on skin improvement including antioxidant effect, collagen synthesis promoting effect, skin fine wrinkle improvement effect, hair growth effect, acne prevention effect and skin irritation relieving effect.

Example 14 Preparation of Lotion Containing Rowan Extract Obtained in Example 1

To 8 g of 95% ethanol, 0.05 g of polypyrrolidone, 0.1 g of oleyl alcohol, 0.2 g of polyoxyethylene monooleate, 0.2 g of fragrance, 0.1 g of paraoxybenzoic acid methyl ester, a small amount of antioxidant and a small amount of pigment are dissolved. . 0.05 g of rowan extract obtained in Example 1 and 5 g of glycerin were dissolved in 85.33 g of purified water, and then the mixed solution was added and stirred to obtain a lotion having a skin improvement effect.

Example 15 Preparation of Latex Containing Rowan Extract Obtained in Example 1

1.2 g of cetyl alcohol, 10 g of squalane, 2 g of vaseline, 0.2 g of paraoxybenzoic acid ethyl ester, 1 g of glycerin monoesteralide, 1 g of polyoxyethylene (20 mole added) monooleate and 0.1 g of fragrance were mixed and dissolved at 70 ° C. , 0.5g of rowan extract obtained in Example 1, 5 g of dipropylene glycol, 2 g of polyethylene glycol-1500, 0.2 g of triethanolamine, and 76.2 g of purified water were heated to 75 ° C. to dissolve it. Both mixtures were emulsified and cooled to obtain an emulsion having an oil-in-water (O / W) type skin improvement effect.

Example 16 Preparation of Cosmetic Solution Containing Rowan Extract Obtained in Example 1

5 g of 95% ethyl alcohol, 1.2 g of polyoxyethylene sorbitan monooleate, 0.3 g of kitulose, 0.2 g of sodium hyaluronate, 0.2 g of vitamin E-acetate, 0.2 g of sodium licorice, 0.1 g of paraoxybenzoic acid ethyl ester, 1g of the rowan extract obtained in Example 1 and a suitable amount of pigment was mixed to obtain a essence having a skin improvement effect.

As described above, the present invention has excellent anti-aging effects such as antioxidant effect, MMP inhibitory effect, MMP expression control effect by UV irradiation, antioxidant effect, collagenase activity control effect, skin wrinkle improvement effect, and after UV irradiation. To provide a cosmetic composition exhibiting an effect of alleviating the skin irritation.

In addition, rowan extract has a whitening effect such as melanin production inhibitory effect, a substance causing blemishes, freckles and skin pigmentation, as well as an antibacterial activity and 5 alpha-reductase which have excellent anti-hair loss and acne prevention effect. Provides an inhibitory effect.

In the above description of the preferred embodiment of the present invention, those skilled in the art will be able to perform various modifications without departing from the gist of the present invention, and such modifications are within the protection scope of the present invention. The protection scope of the present invention should be construed as described in the claims.

Claims (9)

Cosmetic composition characterized in that it comprises Rowan extract of Rowanaceae plant as the main active ingredient. The method of claim 1, The rowan extract is a cosmetic composition, characterized in that it contains 0.001 to 30.0% by weight based on the total weight of the freeze-dried. The method according to claim 1 or 2, The extraction solvent is a cosmetic composition, characterized in that selected from purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane, hexane and mixtures thereof. The method of claim 1, Cosmetic composition, characterized in that it is used for skin whitening. The method of claim 1, Cosmetic composition, characterized in that it is used for skin irritation relief. The method of claim 1, Cosmetic composition, characterized in that used for preventing hair loss. The method of claim 1, Cosmetic composition, characterized in that used for anti-aging. The method of claim 1, Cosmetic composition, characterized in that used for preventing acne. The method of claim 1, Cosmetic composition, characterized in that selected from the formulation of the lotion, gel, water-soluble liquid, cream, essence, oil-in-water (O / W) or water-in-oil (W / O) type, ointment.