KR100728813B1 - Cosmetic composition containing extract of cirsium setidens - Google Patents

Abstract

A cosmetic composition containing Korean thistle extract as a main active ingredient, the composition for external application of the skin containing a sesame extract as a main active ingredient, comprising 0.001 to 30.0% by weight of Korean thistle extract, based on the total weight of the composition .

The external skin composition containing Korean thistle extract as the main active ingredient has excellent antioxidant, anti-wrinkle effect, whitening effect, hair growth effect, acne prevention effect, and stimulating effect. It also has antioxidant activity, human fibroblast activity effect, skin fine wrinkle improvement effect, whitening effect, hair growth effect, acne prevention effect, skin irritation relief effect.

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

Description

Skin external composition containing Korean thistle extract as main active ingredient {COSMETIC COMPOSITION CONTAINING EXTRACT OF CIRSIUM SETIDENS}

The present invention relates to an external preparation composition for skin containing Korean thistle extract as a main active ingredient, and more particularly, an extract prepared from the leaves, stems and roots of the Korean thistle of Cirsium setidens , as an active ingredient. It relates to a skin external preparation composition containing -30.0% by weight, preferably 0.01 to 5% by weight, and excellent in antioxidant effect, anti-wrinkle effect, whitening effect, hair growth prevention, acne prevention effect and skin irritation relieving effect.

Skin aging can be divided into intrinsic chronological and phtoaging [Gilchrest BA: J. Am. Acad. Dermatol., 21, 610-613 (1989). Endogenous aging is a naturally occurring aging phenomenon caused by deterioration of physiological functions of living bodies with increasing age [Braverman IM et al .: J. Invest. Dermatol., 78, 434-443 (1982). Photoaging means that the skin is repeatedly exposed to light to change the appearance or function of the skin [Ridder GM et al .: J. Am. Acad. Dermatol., 25, 751-760 (1991). In addition, skin aging may be caused by the activation of free radicals with ultraviolet rays, stress, disease conditions, environmental factors, wounds, and age, and when this condition is exacerbated, it destroys the antioxidant defense network existing in vivo, cells and Damage to tissues promotes 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, 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. 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. Ultraviolet irradiation also activates these degrading enzymes. 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 skin normally produces melanin to protect the skin from the sun's ultraviolet rays. Melanin is produced from melanocytes, a kind of skin cells, and has a natural screening function that is distributed on the surface of the skin and blocks ultraviolet rays harmful to the human body. There is an enzyme called tyrosinase that is synthesized in melanocytes. Tyrosinase generates dopaquinone via dopa (DOPA) based on an amino acid called tyrosine, and melanin, which is a black pigment, is produced by several stages of oxidative condensation reaction from dopaquinone. Ascorbic acid (JP-A 4-9320), hydroquinone (JP-A-6-192062), kojic acid (JP-A-56-7710), arbutin (JP-A 4-9315) and some Although extracts have tyrosinase inhibitory activity and are used as whitening cosmetics, their use is limited due to poor stability in the prescription system, discoloration due to poor coloration, off-flavor, efficacy at the biological level, unclear effect and safety issues. It's happening. And the inhibitory effect of tyrosinase has been demonstrated, but the effect is low in experiments similar to the actual biologic level.

In addition, androgenetic alopecia is directly dependent on the amount of androgen, which is dependent on androgen hormones. Accordingly, many studies have recently been reported for the prevention and treatment of hair loss through suppression of androgen 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.

When explaining the mechanism of hair loss and acne caused by androgen, 5 alpha-Reductase is present in testosterone-reactive tissues such as sebaceous gland, hair follicle, prostate, and epididymis, and is one of the testosterones. is an enzyme involved in metabolizing testosterone to 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 active.

Recently, in order to reduce skin irritation caused by various chemicals, a lot of attention has been focused on functional cosmetics having functions such as antioxidants, wrinkle reduction, and whitening obtained from natural extracts. 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. For example, US Pat. No. 5,972,341 describes the antiwrinkle effect of a plant of the genus Commiphora, in particular Commiphora mukul extract. Japanese Patent Application Laid-open No. Hei 9-672662 describes seaweed extracts having a hyaluroronidase inhibitory effect. Japanese Patent Application Laid-open No. Hei 10-85905 describes the skin texture improvement effect of Fucoidan extracted from Wakame. Japanese Patent Application Laid-Open No. 2-245087 describes the antioxidant effect of Sargassum genus extract. Japanese Patent Application Laid-Open No. 3-294384 describes an antioxidant composition extracted from the genus Hijikia. Japanese Patent Laid-Open No. 7-10769 describes a extract of Phaeophyceae river having an astringent effect.

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 result of selecting the stems and roots of thistle natural products of the Asteraceae, and extracts from them, to prepare the skin antioxidant effect, collagen synthesis As a result of measuring the effect, skin wrinkle relief effect, whitening effect, hair growth effect, acne prevention effect and skin irritation relief effect, it was found that the efficacy as a cosmetic can be expected.

Korean thistle ( Gundres , Cirsium setidens ) is a perennial herb of Campanula Asteraceae. The root is straight, the stem is branched, the leaf is regenerated, the center is petiole, spiral, elliptic lanceolate, pointed, 15 ~ 35cm long, a little hair on the surface, and the back is white, iron The edges are flat or spiny toothed. The flower is reddish purple, 3 ~ 4cm in diameter, and one head is attached at the end of stem or branch. Flowers bloom in July-October and fruits ripen in November. Young leaves are eaten as herbs and are known to be distributed in Korea.

The present invention, using a Korean thistle extract of the chrysanthemum among a variety of natural products, to confirm the efficacy as a raw material of the external skin preparation, to develop a method of using the same, by using this raw material to prepare a skin external preparation having excellent functionality The purpose is to provide.

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 the natural product of Korean thistle and its cosmetics.

In order to achieve the above object, the present invention provides a composition for external application for skin containing Korean thistle extract of the genus Cirsium genus plant as the main active ingredient.

The present invention also provides a topical skin composition, characterized in that the content of Korean thistle extract is 0.001 to 30.0% by weight based on the total freeze-drying 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 effect of increasing the amount is not economical.

 The Korean thistle extract is extracted by extracting at 70 to 85 ° C. using an extracting solvent, followed by cold soaking, and the resulting suspension is filtrated under reduced pressure or freeze-dried at 50 ° C. or lower.

Korean thistle natural product of the present invention may be used alone or two or three selected from the group consisting of purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane and hexane as an extractant. Solvents are mixed and used. The present invention also provides a skin external composition, which is characterized by extracting Korean thistle natural product using 10-95% ethanol as the extraction solvent.

Formulation of the external composition for skin composition is a basic cosmetic formulation consisting of a lotion, gel, water-soluble liquid, cream, essence, oil-in-water (O / W) and water-in-oil (W / O) type, ointment and oil-in-water or water-in-oil makeup It is selected from color cosmetic formulations consisting of base, foundation, skin cover, lipstick, lip gloss, face powder, two-way cake, eye shadow, cheek color and eyebrow pencils.

In addition, the present invention is characterized in that the topical skin composition is a cosmetic composition or a pharmaceutical composition.

Korean thistle extract can be formulated into pharmaceutical compositions such as capsules, liquids, ointments, patch or sustained-release agent using a pharmaceutically acceptable carrier, pharmacologically acceptable bases, carriers, excipients, binders ( Examples: starch, tragacanth gum, gelatin, molasses, polyvinyl alcohol, polyvinylether, polyvinyl pyrrolidone, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose and carboxymethyl cellulose), grinding agents (e.g., agar, Starch, vellatin powder, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, crystalline cellulose, calcium carbonate, sodium bicarbonate and sodium alginate), lubricants (e.g. magnesium stearate, talc, hydrogenated vegetable oils), colorants and the like. The carriers and excipients include lactose, glucose, sucrose, mannitol, potato starch, corn starch, calcium carbonate, calcium phosphate and cellulose.

In addition to the above, additives such as adjuvants such as stabilizers, dissolution aids, transdermal absorption accelerators, and preservatives may be further added. The pharmaceutical composition prepared as described above may be applied to the skin once or several times daily according to the symptoms, and the application may be adjusted according to the improvement of the symptoms.

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: Korean thistle extract preparation

The dried Korean thistle was refluxed three times for 5 hours with an aqueous 95% (V / V) ethanol solution, cooled, and filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure and lyophilized at 50 ° C. or lower, and thistle was used by using at least one solvent selected from purified water, ethanol, butylene glycol, and propylene glycol to contain 0.001 to 30.0 wt% of the reduced pressure concentrate and lyophilisate. An extract was prepared.

Example 2 Experiment of Measuring Antioxidant Effect Using NBT Method

In order to confirm the antioxidant effect of Korean thistle extract obtained in Example 1, BHT, which is well known as an antioxidant under laboratory conditions, was compared and the antioxidant activity was measured using NBT (Nitro Blue Tetrazolium) 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 effect was calculated by Equation 1, and 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

Sample Name Antioxidative effect(%) Korean thistle extract 0.1% by weight 98% Korean thistle extract 0.01% by weight 85% Korean thistle extract 0.001% by weight 48% BHT 0.1 wt% 89%

Korean thistle extract as shown in Table 1 exhibited an excellent antioxidant effect, it was confirmed that has a superior antioxidant power than BHT.

Example 3: Free radical scavenging activity measurement experiment

In order to measure the antioxidant effect of the Korean thistle extract obtained in Example 1, an antioxidant such as BHT was compared with a sample and the antioxidant activity was measured using the DPPH method.

DPPH method measures the antioxidant activity by reducing power using a free group called DPPH (2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl free radical). 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.

Korean thistle extracts of 0.1%, 0.01% and 0.001% concentration were prepared for DPPH free radical scavenging activity measurement. Extracts of the above concentrations were placed in 96-well plates, respectively, and DPPH prepared in 100 uM ethanol solution was added to obtain a total volume of 200 μl. After leaving it at 37 ° C. for 30 minutes, absorbance was measured at 560 nm. Free radical scavenging activity (%) was calculated by the following equation.

Free radical scavenging activity (%) = 100- (B / A) × 100

A: absorbance of control wells not treated with the extract of the present invention

B: Absorbance of Experimental Wells Treated with Extracts of the Invention

Sample Name Antioxidative effect(%) Korean thistle extract 0.1% by weight 95% Korean thistle extract 0.01% by weight 73% Korean thistle extract 0.001% by weight 58% BHT 0.1 wt% 84%

As can be seen in Table 2, Korea thistle extract was found to have an excellent antioxidant effect than BHT, which is widely used as an antioxidant.

Example 4 In Vitro MMP-1 Inhibition Assay

Substrate metalloproteinase (MMP-1) inhibitory activity was measured in vitro in a screening type biochemical model based on the use of collagen and gelatin conjugated to purified collagenase and its substrate, fluorescein. (EnzChek ™ Gelatinase / Collagenase Kit, Molecule Prove).

Collagenase purified from Clostridium historiescom was supplied in the EnzChek ™ gelatinase / collagenase kit.

The 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 identified as EnzChek® gelatinase. Collagenase kit (Molcula Probes) was used.

Korean thistle extract was dissolved in the reaction buffer and tested at 0.01% by weight, 0.5% by weight, 0.1% by weight. Dilutions of the test extracts were incubated with 25 μg / 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 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.

Sample Name % Inhibition Korean thistle extract 0.1% by weight 83 Korean thistle extract 0.05% by weight 67 Korean thistle extract 0.01% by weight 48 0.2% by weight of green tea extract 71

Korean thistle extract tested at 0.01-0.1% by weight under selected experimental conditions retained dose dependent anticollagenase / gelatinase activity.

As a result of the experiment, as shown in Table 3, 0.1% of Korean thistle extracts inhibited the collagen degradation activity of Clostridium collagenase by 83%, which was superior to the inhibitory effect of green tea extract.

Example 5 Evaluation of Inhibition of MMP-1 Expression by Korean Thistle Extract After UV Irradiation

In this Experimental Example, ELISA was performed to measure the concentration of MMP-1 after UV irradiation and sample addition of Korean thistle extract obtained in Example 1.

Human dermal fibroblasts were irradiated with UVA at an energy of 6.3 J / cm 2 using a UV chamber. UV irradiation amount and incubation time were established through preliminary experiments to maximize the expression of MMP in fibroblasts. The negative control was wrapped in tinfoil to keep the same time in the environment of UVA. UVA emission was measured using a UV radiometer. The cells during the UVA irradiation were intact with the previously dispensed medium, and were irradiated with UVA, exchanged with the medium containing the sample, and then cultured for 24 hours, and the medium was recovered and coated in 96 wells. The primary antibody (MMP-1 (Ab-5) monoclonal antibody) was treated and reacted at 37 ° C. for 60 minutes. After reacting the secondary antibody anti-mouse IgG (whole mouse, alkaline phosphatase conjugated) again for about 60 minutes, the alkaline phosphatase substrate solution (1 mg / ml ρ-nitrophenyl phosphate in diethanolamine buffer solution) was reacted at room temperature for 30 minutes and then Absorbance was measured at 405 nm with a plate reader. As a control, one without addition of a sample was used.

Test group MMP-1 expression inhibition rate (%) Control - Korean thistle extract 0.05% by weight 72 Korean thistle extract 0.01% by weight 41 Retinol 35

As a result, the extract of Korea thistle showed over 72% inhibition rate compared to the control group that did not process the MMP-1 expression induced by UV irradiation, which was significantly superior to the inhibition rate of retinol used as a control.

Example 6: Tyrosinase Activity Inhibition Experiment

Tyrosinase was isolated and purified from mushrooms and purchased from Sigma. Substrate L-tyrosine (Sigma) was dissolved in 0.05M sodium phosphate buffer (pH 6.8) to make a 0.1mg / ㎖ solution was used. Korean thistle extract was used after dissolving it in an appropriate concentration in 0.05M sodium phosphate buffer (pH 6.8). 0.5 ml of L-tyrosine solution was placed in a test tube, and 0.5 ml of Korean thistle extract sample solution was added thereto and left for 10 minutes in a 37 ° C thermostat. Thereafter, 0.5 ml of 200 unit / ml tyrosinase was added to each sample solution, and the mixture was reacted at 37 ° C for 10 minutes. The test tube containing the reaction solution was placed on ice, quenched to stop the reaction, and the spectrophotometer measured absorbance at a wavelength of 475 nm. As a control, 0.5 ml of buffer solution was used instead of the sample.

The tyrosinase activity inhibition rate of each sample was calculated according to Equation 3.

Enzyme Inhibition Rate (%) = 100 ― [(Comparative Absorbance / Control Absorbance)] × 100

Sample Name Tyrosinase inhibition rate (%) Control - Korean thistle extract 0.05% by weight 65% Korean thistle extract 0.01% by weight 49% Arbutin 0.2 wt% 68%

As a result, Korean thistle extract showed more than 65% tyrosinase inhibition rate compared to the control group, which was significantly superior to the inhibition rate of arbutin.

Example 7: Melanin synthesis inhibition experiment using B16F1 melanocytes

In order to confirm the whitening effect of Korean thistle extract obtained in Example 1, the melanin synthesis inhibitory effect on B16F1 melanocytes was measured.

The melanin synthesis 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, and the cells were attached and then 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 number of cells 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 (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 production inhibition rate (%) of B16F1 melanocytes was calculated by the equation (4).

% Inhibition = [(A-B) / A] × 100

A: Melanin amount in wells without sample

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

Sample Name Melanin Synthesis Inhibitory Effect (%) Control - Korean thistle extract 0.05% by weight 39% Korean thistle extract 0.01% by weight 22% Arbutin 0.2 wt% 34%

As a result of testing the melanin synthesis inhibitory effect of B16F1 melanocytes, Korean thistle extract showed melanin production inhibitory effect of 0.05% to 39%, similar to arbutin (Table 6).

Example 8 Inhibitory Effect of Prostaglandin Biosynthesis by UV Irradiation

In this experimental example, to evaluate the inhibitory effect of prostaglandin biosynthesis of Korean thistle extract obtained in Example 1, 5 × 10 ⁴ keratinocytes isolated from human epidermal tissue were placed on a 24-well test plate for 24 hours. While attached. After replacement with medium containing no FBS and incubation for 18 hours, prostaglandin biosynthetic enzymes (prostaglandin E2 synthetase, present in keratinocytes) were treated with aspirin to achieve a final concentration of 50 uM. Or cyclooxygenase (hereinafter referred to as COX) activity was removed. Two hours after aspirin treatment, each well containing keratinocytes was washed twice with PBS and 100 μl of PBS was added to each well. The keratinocytes were irradiated with UV 30 mJ / cm 2 using a UVB lamp (Model: F15T8, UV B 15W, Sankyo Dennki, Japan), and then PBS was removed and each cell was treated with keratinocyte growth media (Clonetics, Biowhittacker, MD, USA) 250 μl was added. The Korean thistle extract to be treated therein was incubated for 16 hours. Prostaglandin inhibitory effect of Korean thistle extract was determined by quantifying PGE2 (prostaglandin E2) biosynthesized for 16 hours by taking an appropriate amount of the culture supernatant. The amount of PGE2 was quantified by enzyme immunoassay, and the results showed that thistle extract effectively inhibited the production of prostaglandins by UV light. In particular, the PGE2 produced is known to be produced mainly by the COX-2 enzyme, and thus it can be seen from this experiment that the Korean thistle extract inhibits the induction or activity of the COX-2 enzyme (Table 7).

Sample Name PGE2 Inhibition Rate (%) (-) UV B control (+) UV B - (+) UV B + Korean thistle extract 0.05% by weight 71% (+) UV B + Korean thistle extract 0.01% by weight 35%

Example 9: Paper Disc Test

This Example was a paper disc test (Paper Disc Test) to confirm the antimicrobial activity of acne bacteria of Korea thistle extract obtained in Example 1. 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. The Korean thistle 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 paper disc having a diameter of 8 mm 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 Korean thistle extract was found to be 19 mm.

Example 10: Evaluation of inhibitory activity of 5-alpha-reductase activity of Korean thistle 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 is incubated. Each hole is cultured after adding 0.1.mμ.Ci of radiolabeled testosterone with 3H (tritium) to determine whether fibroblasts use it. The control group should contain no Korean thistle extract. After 24 hours of incubation, the supernatant was obtained, and steroids were 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)). The radioactivity of the points corresponding to testosterone and dihydrotestosterone was measured by using a densitometer to calculate the conversion rate, and the result was compared with the control (conversion rate when no extract was 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)

Korean thistle extract concentration (%) One 0.1 0.05 0.01 0.005 0.001 0.0001 % Inhibition 100.0 99.0 91.0 84.0 69.4 48.9 34.1

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

Example 11: Hair Growth Effect Test

This example prepared a 30% hydrous ethanol solution containing 2% of the dried thistle extract of Korean thistle extract obtained in Example 1 was used for 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, 30% alcohol solution was applied to each individual to observe the growth state.

Elapsed Days / Sample 5 10 15 Korean thistle extract 0.21 0.87 1.79 ± 0.29 Control 0.07 0.24 0.98 ± 0.24

As a result, it can be seen that Korean thistle extract has an excellent hair growth promoting effect (Table 9).

Example 12 and Comparative Example 1

This Example prepared the cosmetics containing the Korean thistle 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 cooled slowly to prepare a cream (Example 12, 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. The experimental results are described in Table 11 as the ΔR8 value of the cutometer SEM 575, where the R8 value represents the nature of the viscoelasticity of the skin.

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 Aster 2 2 I Korean thistle extract One - Propylene glycol 5 5 Purified water Quantity Quantity Quantity

Note) Unit: Weight%

Experimental product Skin elasticity effect (△ R8) Example 12 0.26 Comparative Example 1 0.11

n = 20, p <0.05

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

Example 13 and Comparative Example 2

This Example prepared a cosmetic containing the Korean thistle extract obtained in Example 1 to evaluate 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 13, Comparative Example 2). 20 experimenters (women aged 20-35 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. 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

Raw material Example 13 Comparative Example 2 Fall fall 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 Korean thistle extract One - Propylene glycol 5 5 Purified water 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 13 Comparative Example 2 Example 13 Comparative Example 2 Example 13 Comparative Example 2 medium 7.12 3.52 2.5 1.7 3.1 2.3

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

In the following, other examples are shown. That is, the lotion, milky lotion and essence containing the Korean thistle extract obtained in Example 1 were prepared in Examples 14 to 16. The lotion, milky lotion and essence containing these thistle extract 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 14 Preparation of Lotion Containing Korean Thistle 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 Korean thistle extract obtained in Example 1 and 5 g of glycerine were dissolved in 85.33 g of purified water, and the mixed solution was added and stirred to obtain a lotion having a skin improvement effect.

Example 15 Preparation of Latex Containing Korean Thistle 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 Korean thistle 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 Korean Thistle 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 Korean thistle extract obtained in Example 1 and an appropriate amount of pigment were mixed to obtain a essence having a skin improvement effect.

As described above, the Korean thistle 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, and fine wrinkle improvement effect and relieve skin irritation caused by UV irradiation. It works.

In addition, Korean thistle extract has a whitening effect such as melanin production inhibitory effect, which is a substance causing blemishes, freckles and skin pigmentation.

In addition, thistle extract of the Korean thistle shows excellent antibacterial and anti-acne bactericidal activity and 5 alpha-reductase inhibitory effect.

Therefore, the cosmetic composition such as the lotion, cream, milky lotion, pack, powder, etc. containing the extract of Korean thistle has antioxidative effect, collagen degrading activity control effect, skin fine wrinkle improvement effect, whitening, hair growth, acne prevention, skin irritation relief effect, etc. Efficacy as an excellent skin external preparation.

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)

delete delete delete delete Skin thistle extract 0.001 ~ 30.0% by weight and the external composition for skin, characterized in that it is used for acne prevention. Skin thistle extract contains 0.001 ~ 30.0% by weight and is used for inhibiting skin damage or alleviating skin irritation and whitening by ultraviolet rays. Skin thistle extract containing 0.001 ~ 30.0% by weight and is used for hair damage prevention, hair growth composition. delete