KR20070016261A - Cosmetic composition containing an extract of ligularia fischeri - Google Patents

Abstract

The present invention relates to a cosmetic composition containing a bear extract as an active ingredient, more specifically, antioxidant and anti-wrinkle, characterized in that it contains 0.001 to 30.0% by weight of the extract of the stem and root of the bear odor of Campanula Asteraceae. It relates to a cosmetic composition excellent in the effect, whitening effect, hair growth effect, acne prevention effect, stimulation alleviating effect. Bear extract has not only antioxidant activity but also human fibroblast activity effect, skin fine wrinkle improvement effect, whitening effect, hair growth effect, acne prevention effect, skin irritation effect, so that various functional cosmetics can be prepared using this substance.

Bear extract, anti-aging cosmetics, anti-wrinkle effect, whitening effect, hair growth effect, acne prevention effect, skin irritation relieving effect, skin external preparation.

Description

Cosmetic composition containing bear extract as an active ingredient {COSMETIC COMPOSITION CONTAINING AN EXTRACT OF LIGULARIA FISCHERI}

The present invention relates to a cosmetic composition containing a bear odor extract as an active ingredient, more specifically 0.001 to 30.0 weight of the extract prepared from the natural products of the leaves, stems and roots of Ligularia fischeri of Campanula Asteraceae. It relates to a cosmetic composition excellent in antioxidant, anti-wrinkle effect, whitening effect, hair growth effect, acne prevention effect, skin irritation effect characterized in that it contains%, preferably 0.01 to 5% by weight It is to provide an excellent cosmetic composition with an anti-aging effect, anti-wrinkle effect, whitening effect, hair growth prevention, acne prevention effect and irritation relief effect as an extract prepared from natural products.

Human skin has various physical and chemical changes during the aging process, and its causes are classified into internal aging and photo-aging. In other words, ultraviolet rays, stress, disease conditions, environmental factors, wounds, and age can be caused by the activation of free radicals, if this condition is intensified, destroys the antioxidant defenses in vivo, damage cells and tissues 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 proteins is caused by severe cuts in connective tissues such as collagen, hyaluronic acid, elastin, proteoglycan, fibronectin, which severely inhibit inflammatory reactions and elasticity of the skin. This leads to a situation of reduced immune function.

Therefore, it protects the cell membrane by eliminating the free radicals generated by metabolic process of the body, UV irradiation, and inflammatory reactions.Also, damaged cells should be regenerated by active metabolism to proliferate the skin. You can recover quickly and maintain healthy skin. 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 metalloproteinase, an enzyme that degrades collagen ( MMP) expression is promoted, the elasticity of the skin is reduced, wrinkles are formed. In addition, such enzymes may be 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. Until now, most of the raw materials used as materials for cosmetics simply inhibited enzyme activity. Therefore, we tried to regulate the expression and activity of MMP expression induced by intracellular aging and photoaging.

Next is skin change by pigmentation. The pigments that affect skin color include melanin, melazoids, carotene, and hemoglobin, the most important of which is melanin, which is the most important factor affecting biosynthesis. 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 active oxygen species, but sometimes melanin itself generates free radicals, and other substances are reduced or oxidized by catechol or quinone in 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. Although it is used as a whitening cosmetic because it has inhibitory activity, its use is restricted due to poor stability in cosmetic formulations, occurrence of odor, efficacy at the biological level, unclear effect and carcinogenic substance, and safety problems. There is a situation.

In addition, male alopecia is male hormone-dependent, and thus has a direct relationship with the amount of male hormones. Accordingly, many studies have recently been reported for the prevention and treatment of hair loss through 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 impulse, skeletal muscle increase, male external genitalia, scrotum growth, spermatogenesis, etc., and dihydrotestosterone is involved in tissues such as acne, sebum increase, hair loss, and prostatic hyperplasia. (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 actively underway.

In order to solve the problem of the skin, a lot of cosmetics using natural products have recently 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, by selecting the stems and roots of the natural bear odor and extracts from them, the skin antioxidant effect, collagen synthesis effect, As a result of measuring skin wrinkle relief, whitening effect, hair growth effect, acne prevention effect and skin irritation relaxation effect, it was found that it can be expected to be effective as a cosmetic.

Ligularia fischeri is a dicotyledonous plant of the Asteraceae. It is a perennial herb, stem is 1 ~ 2m high, the root stem is thick, and there are coarse white fine hairs on the lower part and short hairs on the upper part. The leaves above the roots have long petioles. The leaves are heart-shaped and have serrated edges. The leaf front is green and the back is pale green. The stem has three leaves, the bottom leaf is small and encloses the petiole. The leaves at the top are much smaller, the petioles are shorter, and the bottoms are wider, like leafy leaves. It is a gunshot inflorescence, which blooms and branches from the bottom. Tubal is brown or brown purple, fruit is achene (瘦果), columnar, 7-11mm long, with vertical lines. Flowers bloom in July-September and fruits ripen in October. Young leaves are eaten as herbs and are known to be distributed in Korea, Japan, China and Sakhalin.

The present invention selects an extract prepared from the natural odor of the lily family of several natural products, to confirm the efficacy as a raw material of the skin external preparation, to develop a method of use thereof, by using this raw material to provide a skin external preparation having a predetermined functionality It is an object to provide a method which can be manufactured.

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.

It is also an object of the present invention to provide a natural extract exhibiting a whitening effect by ultraviolet irradiation and the like to alleviate skin irritation by an inflammatory mediator when applied to skin cosmetics.

It is also an object of the present invention to provide a natural extract exhibiting a 5-alpha-reductase inhibitory effect having an excellent hair loss prevention effect and acne prevention effect.

In addition, the present invention is to provide a method for producing a cosmetic using the extract prepared from natural products of the bear odor and its cosmetics.

In order to achieve the above object, the present invention provides a cosmetic composition containing a bear extract of Asteraceae as an active ingredient.

In addition, the bear extract is characterized in that it contains 0.001 to 30.0% by weight based on the whole 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, the degree of increase in the effect of increasing the content is insignificant and thus it is not economical.

The bear extract is extracted by cold extraction by using an extracting solvent at room temperature of 70 ~ 85 ℃, and the resulting suspension is filtered to obtain a liquid obtained by further concentrating under reduced pressure or freeze drying at 50 ℃ or less. The extraction solvent may be used alone or in combination of two or three solvents selected from the group consisting of purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane and hexane. .

In addition, the bear extract is used for anti-aging, whitening, hair damage prevention, hair growth, acne prevention, skin damage suppression or irritation relief by ultraviolet light.

Formulation of the cosmetic composition is a lotion, gel, water-soluble liquid, cream, essence, oil-in-water (O / W) and water-in-oil (W / O) type emulsion, ointment base cosmetic formulation and oil-in-water or water-in-oil makeup base , Foundation, skin cover, lipstick, lip gloss, face powder, two-way cake, eye shadow, teak color and eyebrow pencils.

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 Bear Extract

After dipping, the dry bear odor was refluxed three times for 5 hours with 95% (V / V) ethanol aqueous solution, and the mixture was cooled and then filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure and freeze dried at 50 ° C or lower. Bearish extract was prepared by dispersing / dissolving at least one solvent selected from purified water, ethanol, butylene glycol, and propylene glycol so as to contain 0.001 to 30.0 wt% of the reduced pressure concentrate and the lyophilized product.

Example 2 Experiment of Measuring Antioxidant Effect Using NBT Method

In this example, to determine the antioxidant effect of the bear extract obtained in Example 1 was compared to other antioxidants, namely retinol and BHT under laboratory conditions, 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 produced by xanthine and xanthine oxidase. This active oxygen reacts with Nitro Blue Tetrazolium (NBT) to measure the active oxygen scavenging rate by measuring the blue color produced by this 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)] × 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 addition of sample solution

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

As a result, as shown in Table 1, it was confirmed that the excellent antioxidant effect in all the samples tested at 0.1% concentration. Bear extract had excellent antioxidant effects similar to retinol and BHT.

Sample Name Treatment Concentration (%) Antioxidative effect(%) Bear extract (Example 1) 0.1 94 Retinol 0.1 93 BHT 0.1 90

Example 4 Experiment for Measuring Antioxidant Effect Using DPPH Method

In this example, the antioxidant activity of the extract of Goji odor obtained in Example 1 was measured using a DPPH method using a green tea extract and an antioxidant such as vitamin E as a comparative sample under laboratory conditions.

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)] × 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, the bear extract had better antioxidant effect than green tea extract and vitamin E at 0.01% concentration.

Sample Name Treatment Concentration (%) Antioxidative effect(%) Bear extract (Example 1) 0.01 83 Green Tea Extract 0.01 45 Vitamin E 0.01 43

Example 5: in vitro MMP-1 inhibition assay

Matrix 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 ™) Tinase / collagenase kit, by Molar Prov.). 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 EnzChek (TM) gelatinase. Collagenase kit (Molcula Probes) was used. The protonated and non-extracted extracts were 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. In each experimental condition, a blank, hereinafter referred to as '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, bear extracts tested at 0.04-4% (v / v) under selected experimental conditions retained anti-collagenase / gelatinase activity in a dose dependent manner.

As shown in Table 3, the Gom-odor extract inhibited 83% of collagen degrading activity of Clostridium collagenase when treated with 0.04%, which was better than the inhibitory effect of green tea extract.

 Experimental Sample % Inhibition Treatment concentration 0.02 0.04 Bear extract (Example 1) 55 83 Retinol 0 5 Green Tea Extract 52 58

Example 6 Evaluation of Inhibition of MMP-1 Expression by Bear Extract after UV Irradiation

This Example was subjected to ELISA to measure the concentration of MMP-1 after UV irradiation and sample addition of the bear 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. Measure the absorbance at 405 nm. As a control, the one without addition of the sample is used.

In the case of MMP-1 expression induced by UV irradiation, the bear extract extract showed more than 20% inhibition rate compared to the control group without treatment, which is superior to the inhibition rate of retinol used as a control (Table 4).

Test group Treatment concentration (%) MMP-1 expression inhibition rate (%) Control - - Bear extract (Example 1) 0.1 27 Retinol 0.1 18

Example 7: Experiment for measuring 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 bear 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 the 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 × 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. Melanin was extracted by adding 1N NaOH (10% DMSO) to the cell filtrate obtained by centrifugation (3,000 rpm, 10 minutes), and then measured the absorbance of melanin at 405 nm with a microplate reader. Percent inhibition of production 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] × 100

A: Melanin amount in wells without sample

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

Test results of melanin production inhibitory effect of B16F1 melanocytes showed an IC ₅₀ value of 0.08%, which was similar or superior to conventional whitening agents such as hydroquinone, arbutin, oil-soluble licorice extract and lettuce extract (Table 5). .

Sample Name Treatment concentration (%) Inhibitory effect of melanin synthesis (IC ₅₀ ) Bear 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 8: Cytotoxic Alleviation Effect by UV Irradiation

This example was carried out to evaluate the alleviation effect of cytotoxicity by ultraviolet irradiation of the bear extract obtained in Example 1. Fibroblasts (fibroblasts) were placed in 24-well test plates 1 × 10 ⁵ and attached 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. Herein, the treatment was performed for 24 hours after treatment of the extract and Visage extract to be evaluated. After 24 hours, the medium was removed, and 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)] × 100

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

Bt: 565 nm absorbance of wells that developed and reacted wells without sample

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

% Reduction of cytotoxicity by UV light = [1- (St-Bo) / (Bt-Bo)] × 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 viability of wells treated with UV

As a result of the experiment, bear extract was found to effectively protect against cytotoxicity by ultraviolet light by alleviating cytotoxicity by 55% at 0.013% concentration. Through the experiment it can be seen that effectively protects against cell damage caused by ultraviolet rays at low concentrations (Table 6).

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

Example 9 Inhibitory Effect of Inflammatory Cytokine Expression by UV Irradiation

This Example is to evaluate the inhibitory effect of the inflammatory cytokine expression expressed by ultraviolet irradiation of the bear extract obtained in Example 1 Fibroblasts isolated from human epidermal tissue (5 ×) on a 24-well test plate 10 ⁴ each was added 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 ultraviolet light 10 mJ / cm 2 using an ultraviolet B (UVB) lamp (Model: F15T8, UV B 15W, Sankyo Dennki Co., Japan), after which PBS was removed and cell culture medium (FMEM was not added to DMEM). Medium) 350 μl was added. After treatment with the bear odor extract to be evaluated it was incubated for 5 hours. 150 μl of the culture supernatant was taken to quantify IL-1α to determine the inhibitory effect on the inflammatory cytokine expression of the extract from Goji. 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)] × 100

Bo: IL-1α production in wells without UV irradiation

Bt: IL-1α production in wells that were not irradiated with UV light

St: IL-1α production amount of wells that were irradiated with UV light

Experimental results show that bear extract inhibits 58% of the inflammatory cytokine-induced inflammatory cytokine IL-1α at a concentration of 0.013%, effectively preventing the occurrence of inflammation caused by UV rays (Table 7).

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

Example 10: Paper Disc Test

This Example was a paper disc test (Paper Disc Test) to confirm the antimicrobial activity of the bear extract obtained in Example 1 against acne bacteria. First, in order to activate propionibacterium acnes, a skin flora of the cause of acne, it was preincubated 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. After diluting the bear extract obtained in Example 1 with 12% (W / v) in a 95% ethanol aqueous solution and dropping 50 μl each on a 8 mm diameter paper disc, it was placed on the solid medium prepared above, followed by an anaerobic culture at 35 ° C. 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 bacterium growth inhibitory ring of Bear extract was found to be 22 mm.

Example 11: Evaluation of inhibitory activity of 5-alpha-reductase activity of bear extract

5 alpha-reductase activity inhibitory 5 alpha-reductase used in the experiment was used for the enzyme produced by the fibroblasts derived from the foreskin.

Fibroblasts are inoculated so that 10000 cells per microplate hole are incubated. Each hole is cultured after adding 0.1.mμ.Ci of radiolabeled testosterone with 3H (tritium) to determine whether fibroblasts use it. As a control, no bear extract is added. After 24 hours of incubation, the supernatant is obtained and steroids are obtained with 1 ml of ethyl acetate-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)

Experimental results show that bear extract has a 5 alpha-reductase inhibitory effect (Table 8).

Bear 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 12 Hair Growth Effect Test

This example prepared a 30% hydrous ethanol solution containing 2% of the Bear extract dry extract obtained in Example 1 was used in the test. The hair growth effect test was performed using a mouse (ICR), 47-53 days of age, to remove the back hair, and to select the clean back area, using 10 animals per material group. Each ml was applied. 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 of the experiment, it can be seen that the bear extract extract has an excellent hair growth promoting effect (Table 9).

Elapsed Days / Sample 5 10 15 Bear Extract 0.27 0.96 1.92 ± 0.43 Control 0.08 0.26 1.01 ± 0.23

Example 13 and Comparative Example 1

This Example prepared the cosmetics containing the bear odor extract obtained in Example 1 to evaluate the skin elasticity improvement effect in 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 it is cooled slowly to prepare a cream (Example 13, Comparative Example 1). Twenty test subjects (20-35 year old female) were applied to the cream prepared in Example 13 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. 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 improving effect of the experimenter who applied the cream containing the bear odor extract was excellent.

Raw material Example 13 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 part a) Alcohol ester 3 3 Glyceryl Monostearic Acid Aster 2 2  I Bear Extract One - Propylene glycol 5 5 Quantity Quantity Quantity

Note) Unit: Weight%

Experimental product Skin elasticity effect (△ R8) Example 13 0.24 Comparative Example 1 0.12

n = 20, p <0.05

Example 14 and Comparative Example 2

This Example prepared the cosmetics containing the bear odor extract obtained in Example 1 and evaluated the skin whitening effect in a comparative experiment with Comparative Example 2 in humans.

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 14, Comparative Example 2). 20 experimenters (women aged 20 to 35 years) were applied to the right side of the face with the cream prepared in Example 14 and the left side of the face with the cream prepared in Comparative Example 2 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 the bear 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 Aster 2 2  I Bear 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 the Subject Example 14 Comparative Example 2 Example 14 Comparative Example 2 Example 14 Comparative Example 2 medium 5.03 2.03 2.7 1.9 2.5 1.7

Hereinafter, as other examples, the lotion, milky lotion and essence containing the odor extract obtained in Example 1 were prepared in Examples 15 to 17. The lotion, milky lotion and essence containing these extracts showed excellent effects on skin improvement such as antioxidant effect, collagen synthesis promoting effect, skin fine wrinkle improvement effect, hair growth effect, acne prevention effect and skin irritation relieving effect.

Example 15 Preparation of Lotion Containing the Bear 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 bear extract obtained in Example 1 and 5 g of glycerin were dissolved in 85.33 g of purified water.

Example 16: Preparation of Latex Containing Bear Extract Extract Obtained in Example 1

1.2 g of cetyl alcohol, 10 g of squalane, 2 g of petrolatum, 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. , 0.5 g of the bear 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 then cooled to obtain an emulsion having an oil-in-water (O / W) skin-improving effect.

Example 17 Preparation of Cosmetic Liquid Containing Bear Extract 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, 1 g of the bear extract obtained in Example 1 and an appropriate amount of pigments were mixed to obtain a cosmetic liquid having a skin improvement effect.

As described above, the bear extract according to the present invention provides an excellent anti-aging effect such as antioxidant effect, MMP inhibitory effect, MMP expression control effect by UV irradiation, fine wrinkle improvement effect and alleviates skin irritation caused by UV irradiation. The effect was shown.

In addition, the bear extract extract whitening action, such as melanin production inhibitory effect, which causes blemishes, freckles and skin pigmentation, excellent hair loss prevention effect, acne bacteria antibacterial activity with anti-acne effect, and 5 alpha-reductase inhibitory The effect was shown.

Therefore, the cosmetic composition such as a lotion, cream, milky lotion, packs, powders, etc. containing the extracts of bears have excellent antioxidative and collagen degrading activity, skin wrinkle improvement, whitening, hair growth, acne prevention, skin irritation It can be seen that it has an external skin 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 (8)

Cosmetic composition containing the extract of Bear odor of Asteraceae as an active ingredient. The method of claim 1, Cosmetic composition containing a bear odor extract as an active ingredient, characterized by containing 0.001 to 30.0% by weight of the bear odor extract based on the total weight of the composition. The method of claim 1, The bear extract is extracted by cold extraction by extraction at room temperature of 70 ~ 85 ℃ using an extracting solvent, the resultant obtained by filtering the liquid suspension obtained by filtration under reduced pressure or freeze drying at 50 ℃ or less. Cosmetic composition containing the as an active ingredient. The method of claim 3, The extraction solvent may be used alone or selected from the group consisting of purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane and hexane, or a mixture of two or three solvents. Cosmetic composition containing a bear extract as an active ingredient, characterized in that. The method according to any one of claims 1 to 4, The cosmetic composition is a cosmetic composition containing a bear extract as an active ingredient, characterized in that for acne prevention, skin aging prevention. The method according to any one of claims 1 to 4, The cosmetic composition is a cosmetic composition containing a bear odor extract as an active ingredient, characterized in that the skin damage by ultraviolet rays or skin irritation relief, whitening. The method according to any one of claims 1 to 4, The cosmetic composition is a cosmetic composition containing a bear odor extract, characterized in that for preventing hair damage, hair growth as an active ingredient. The method according to any one of claims 1 to 4, Formulation of the cosmetic composition is a lotion, gel, water-soluble liquid, cream, essence, oil-in-water (O / W) and water-in-oil (W / O) type emulsion, ointment base cosmetic formulation and oil-in-water or water-in-oil makeup base Contains foundation extract, skin cover, lipstick, lip gloss, face powder, two-way cake, eyeshadow, teak color and eyebrow pencils as one of the color cosmetic formulations selected from the group as an active ingredient Cosmetic composition.