KR20070021585A - Cosmetic composition containing extract of sedum sarmentosum - Google Patents

Abstract

The present invention is an external skin composition containing dolmul extract as the main active ingredient, the external skin composition containing dolmul extract as a main active ingredient, characterized by containing 0.001 to 30.0% by weight of the dolmul extract relative to the total weight of the composition Is provided.

Skin external composition containing dolna extract as the main active ingredient is excellent in antioxidant effect, anti-wrinkle effect, whitening effect, hair growth effect, acne prevention effect, stimulating alleviation 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, and moisturizing effect.

Stone sprout extract, anti-aging, anti wrinkle, whitening, hair growth, acne prevention, skin irritation relief.

Description

Skin external composition containing dolnamul extract as the main active ingredient {COSMETIC COMPOSITION CONTAINING EXTRACT OF SEDUM SARMENTOSUM}

The present invention relates to an external composition for skin containing sedum extract as a main active ingredient, more specifically, 0.001 to 30.0% by weight of an extract prepared from natural products of leaves, stems and roots of sedum ( Sedum sarmentosum ), Preferably, the present invention relates to a skin external preparation composition excellent in antioxidant, anti-wrinkle effect, whitening effect, hair growth effect, acne prevention effect, and skin irritant effect, which contains 0.01 to 5% by weight. Extract prepared from to provide an anti-aging effect, anti-wrinkle effect, whitening effect, hair growth prevention, acne prevention effect and irritation relief effect excellent skin external composition.

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.

Next, 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 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 active.

Next, the stratum corneum, the outermost layer of the skin, has a barrier against moisture evaporation and absorption control of the skin and penetration of external substances such as chemicals, toxic substances and bacteria. The stratum corneum is filled with keratinocytes and intercellular lipids and forms a lamellae. The stratum corneum of the skin contains about 20% water, and it is well known that the moisture content is an important factor in determining the flexibility and moisture of the skin. The ability to retain moisture in the stratum corneum is controlled by a natural moisturizing factor (NMF) consisting of sebum, amino acids, lactic acid, urea and inorganic salts. The development of highly moisturizing substances is one of the important research tasks in cosmetics. It is the cause of reducing the water retention capacity of the stratum corneum by promoting the cleavage and low molecular weight of hyaluronic acid, which plays a major role in the water retention capacity of the skin. In general, when the moisture of the stratum corneum is less than 10%, the physiological action of the skin is lowered, the skin loses its original function, becomes rough, and causes various skin problems. Therefore, the use of moisturizers to prevent drying of the skin is almost essential in cosmetics. Currently used moisturizers include polyols such as glycerin, propylene glycol and butylene glycol, natural moisturizing factors such as amino acids, pyrrolidone carbonates and lactates, biopolymers such as hyaluronic acid salts, chondroitin sulfates and chitosan, etc. There is this. Some of these are moisturizers that, although exhibiting excellent moisturizing properties, are synthetic compounds or animal-derived moisturizers, which, when used in excess, can not exclude skin irritation or contamination-induced infections.

Recently, in order to reduce skin irritation caused by various chemicals, much attention has been focused on functional cosmetics having functions such as antioxidant, wrinkle relief, whitening, and moisturizing. 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 Phaeophyceae extract having an astringent effect.

Thus, the present inventors have studied the possibility of applying to cosmetics in a variety of natural products that are not known until now, by selecting the stems and roots of the Roots of the Roots and Roots natural products and preparing extracts from them, the skin antioxidant effect, collagen synthesis effect As a result of measuring skin relieving effect, whitening effect, hair growth effect, acne prevention and skin irritation effect, and moisturizing effect, it was found that it can be expected to be effective as a cosmetic.

Sedum sarmentosum is a perennial herb with rosacea sedum, with stems extending laterally, with roots at each node, and peduncles standing upright, 15 cm high. The leaves are usually 3 turnsed, without petioles, long oval or lanceolate. The leaves are pointed at both ends and the edges are flat. The flower is yellow and blooms in August ~ September. 5 petals are lanceolate, pointed, longer than calyx. Fruits are goldol (利 咨 果) and there are five carpels (心 皮). Young stems and leaves are soaked in kimchi and light Hansoon is made of herbs. It is known to be distributed in mountains throughout Korea, Japan, and China.

An object of the present invention is to provide a composition for external application for skin comprising sedum extract with sedum among various natural products as a main active ingredient.

It is also an object of the present invention to provide a natural extract that can be applied to skin cosmetics and exhibits excellent anti-aging effects such as antioxidant effects, collagen synthesis effects, and skin wrinkle prevention effects when contained in cosmetics.

It is also an object of the present invention to provide a skin external preparation composition containing as a main active ingredient a natural extract exhibiting a whitening effect by ultraviolet irradiation and the like to reduce skin irritation by an inflammation mediator.

It is also an object of the present invention to provide a skin external preparation composition containing as a main active ingredient a natural extract showing a 5 alpha-reductase inhibitory effect having an excellent hair loss prevention effect and acne prevention effect.

In order to achieve the above object, the present invention provides a topical skin composition containing a natural stone as a main active ingredient.

In addition, the total weight of the skin external preparation composition, it contains 0.001 to 30.0% by weight of dolmul extract. 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 content is not so small that it is not economical.

In addition, the dolmul extract is extracted by cold extraction at room temperature of 70 ~ 85 ℃ using an extracting solvent, the resulting suspension is filtered and obtained by further concentrating under reduced pressure or freeze drying at 50 ℃ or less.

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.

The dolnamul 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 (e.g. : Starch, tragacanth gum, gelatin, molasses, polyvinyl alcohol, polyvinyl ether, polyvinyl pyrrolidone, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose and carboxymethyl cellulose), grinding agents (e.g., agar, starch) , Bellatin powder, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, crystalline cellulose, calcium carbonate, sodium bicarbonate and sodium alginate), lubricants (e.g. magnesium stearate, talc, hydrogenated vegetable oil), 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 a day depending on the symptoms and may adjust the amount applied according to the improvement of 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: Preparation of Sedum Sprout Extract

The dried and dried dolmuls 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 lyophilized at 50 ° C or lower. The sedum 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 order to confirm the antioxidant effect of the sedum extract obtained in Example 1 was compared with other antioxidants, that is, retinol and BHT under laboratory conditions, 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 removing the active oxygen by the test substance, 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 Treatment Concentration (%) Antioxidative effect(%) Stone sprout extract (Example 1) 0.1 93 Retinol 0.1 91 BHT 0.1 89

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. Sedum extracts had excellent antioxidant effects similar to retinol and BHT.

Example 3: Free radical scavenging activity measurement experiment

In this Example, the antioxidant activity of the sedum extract obtained in Example 1 was measured using 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 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.

In order to measure DPPH free radical scavenging activity, sedum extracts of 0.1%, 0.01% and 0.001% concentrations were prepared. 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 Treatment concentration (%) Antioxidative effect(%) Stone sprout extract (Example 1) 0.01 71 Green Tea Extract 0.01 48 Vitamin E 0.01 45

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

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. Reaction buffer consisting of DQ-collagen purified from porcine skin and conjugated to fluorescein with 0.05M Tris-HCl, 0.15M NaCl, 5mM CaCl2 and 0.2mM sodium azide (pH 7.6) was found in EnzChek® gelatinase / Collagenase kit (Molcula Probes) was used. Sedum extract was dissolved in the reaction buffer. It was tested at 0.01%, 0.5%, 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. 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. In conclusion, the sedum extracts tested at 0.01 to 0.1% by weight under selected experimental conditions retained dose dependent anticollagenase / gelatinase activity.

Experimental Sample Inhibition rate 0.02 (treatment concentration (%)) 0.04 (treatment concentration (%)) Stone sprout extract (Example 1) 58 76 Retinol 0 10 Green Tea Extract 57 68

As shown in Table 3, the dolmul extract inhibited 76% of the collagen degrading activity of Clostridium collagenase when treated with 0.04%, which is superior to the inhibitory effect of green tea extract.

Example 5: Evaluation of the inhibition of MMP-1 expression by sedum 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 the sedum 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 in the previously dispensed medium and 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.

In the case of MMP-1 expression induced by UV irradiation, the sedum extract showed more than 54% inhibition rate compared to the control group without treatment, which was significantly superior to the inhibition rate of the retinol used as a control.

Test group Treatment concentration (%) MMP-1 expression inhibition rate (%) Control - - Stone sprout extract (Example 1) 0.1 54 Retinol 0.1 29

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. Sedum extract was used after dissolving 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 dolmul 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 Absorption / Control Absorbance)] × 100

Sample Name Tyrosinase inhibition rate (%) Control - Sedum Extract 0.05% by weight 61% 0.01% by weight sedum extract 34% Arbutin 0.2 wt% 65%

Test Results As shown in Table 5, the dolmul extract showed a similar tyrosinase inhibitory activity to arbutin.

Example 7: Melanin synthesis inhibition experiment using B16F1 melanocytes

In order to confirm the whitening effect of the dolmul 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 x 10 ⁶ per well, and the cells were attached and 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] x 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 - Sedum Extract 0.05% by weight 31% 0.01% by weight sedum extract 18% Arbutin 0.2 wt% 35%

As a result of testing the melanin synthesis inhibitory effect of B16F1 melanocytes, dolmul extract showed a melanin production inhibitory effect of 0.05% to 31%, showing a similar production inhibitory effect to arbutin (Table 6).

Example 8 Inhibition of Prostaglandin Biosynthesis by UV Irradiation

In this experimental example, to evaluate the inhibitory effect of prostaglandin biosynthesis of the sedum extract obtained in Example 1, 5 × 10 ⁴ keratinocytes isolated from human epidermal tissue were placed in a 24-well test plate for 24 hours. 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 prepared with keratinocyte growth media (Clonetics, Biowhittacker, MD, USA) 250 μl was added. After treating the sedum extract to be evaluated here, the culture was carried out for 16 hours. An appropriate amount of the culture supernatant was taken to quantify biosynthesized PGE2 (prostaglandin E2) for 16 hours to determine the prostaglandin inhibitory effect of sedum extract. The amount of PGE2 was quantified using enzyme immunoassay, and experimental results showed that sedum extract effectively inhibited the production of prostaglandins by UV light. Particularly, the produced PGE2 is known to be mainly produced by the COX-2 enzyme, and thus it can be seen that the dolmen extract inhibits the induction or activity of the COX-2 enzyme through the experiment (Table 7).

Sample Name PGE ₂ Inhibition Rate (%) (-) UV B control (+) UV B - (+) UV B + Sedum Extract 0.05% by weight 46% UV B + Sedum Extract 0.01% by weight 21%

Example 9: Paper Disc Test

This Example was a paper disc test (Paper Disc Test) to check the antibacterial activity of the dolmul extract obtained in Example 1 against acne bacteria. First, in order to activate propionibacterium acnes, a dermatological 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 dolmen extract obtained in Example 1 with 12% (W / v) in an aqueous 95% ethanol solution and dropping 50 μl each on a 8 mm diameter paper disc and placing it on a solid medium prepared above, this was 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 bacterial growth inhibitory ring of dolmen extract was found to be 15 mm.

Example 10: Evaluation of inhibitory activity of 5-alpha-reductase activity of sedum 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, do not contain sedum extract. 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)). 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 Evaluation Method

% Inhibition = [A-B] / [A] × 100

A = conversion of testosterone to dihydrotestosterone (without extract)

B = conversion of testosterone to dihydrotestosterone (extract added)

Sedum extract concentration (%) One 0.1 0.05 0.01 0.005 0.001 0.0001 % Inhibition 100.0 96.0 89.0 78.0 57.1 41.3 30.8

Experimental results confirmed that the sedum extract has a 5 alpha-reductase inhibitory effect.

Example 11: Hair Growth Effect Test

This example prepared a 30% hydrous ethanol solution containing 2% dry stone extract 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 hairs, and the back area was selected to be clean. 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.

Elapsed Days / Sample 5 10 15 Sedum extract 0.18 0.59 1.54 ± 0.36 Control 0.09 0.21 0.99 ± 0.16

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

Example 12 and Comparative Example 1

This Example prepared a cosmetic containing the dolmul 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 below as the ΔR8 value of the cutometer SEM 575, where the 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 sedum 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 part a) Alcohol ester 3 3 Glyceryl Monostearic Acid Aster 2 2 I Sedum extract One - Propylene glycol 5 5 Purified water Quantity Quantity Quantity

Note) Unit: Weight%

Experimental product Skin elasticity effect (△ R8) Example 12 0.25 Comparative Example 1 0.12

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 sedum extract was excellent.

Example 13 and Comparative Example 2

In this embodiment, the cosmetics containing the dolmul extract obtained in Example 1 was prepared and evaluated for the effect of improving skin wrinkles and skin beauty by performing a comparative experiment 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 (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 2 on the left side of the face, twice a day for 3 consecutive months.

After completion of the experiment, the skin wrinkle relief effect is collected before and after using the product for 2 months by using a silicone resin copy (replica) around the face of the eye, and comparing the state of the eye wrinkles with a skin fine wrinkle device and a skin image analyzer.

Raw material Example 13 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 part a) Alcohol ester 3 3 Glyceryl Monostearic Acid Aster 2 2 I Sedum extract One - Propylene glycol 5 5 Purified water Quantity Quantity Quantity

Note) Unit: Weight%

Table 13 compares the average eye wrinkle depth and wrinkle reduction rate of the experimenter using the cream prepared in Example 13 with the experimenter using the cream of Comparative Example 2. As shown in Table 13, it can be seen that the skin wrinkle improvement effect is excellent in the skin around the facial eye of the experimenter applying the cream containing the sedum extract.

Example 13 Comparative Example 2 Before use 2 months later Before use 2 months later Average fine wrinkle depth 0.194 0.167 0.191 0.187 Wrinkle Reduction (%) 13.92% 2.09%

n = 20, p <0.01

Example 14 and Comparative Examples 3-6

This Example prepared the cosmetics containing the dolmul extract obtained in Example 1 and evaluated the skin moisturizing effect in a comparative experiment with Comparative Examples 3 to 6 in humans.

The cosmetics used in the comparative experiments are in the form of a cream, the composition of which is shown in Table 14. First, the phase b) recorded in Table 14 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 it cools gradually, and creams (Example 14, Comparative Examples 3-6) are manufactured. Example 20 and Comparative Examples 3-6 were apply | coated to the lower part about 20 experimenters, and after about 90 minutes, (DELTA) Hydration was evaluated by the Corneometer (CM 820) for the moisture retention ability before and after application | coating.

Raw material Example 14 Comparative Example 3 Comparative Example 4 Comparative Example 5 Comparative Example 6    end Stearyl alcohol 8 8 8 8 8 Stearic acid 2 2 2 2 2 Stearic cholesterol 2 2 2 2 2 Squalane 4 4 4 4 4 2-octyldodecyl alcohol 6 6 6 6 6 Polyoxyethylene (25 mol part a) Alcohol ester 3 3 3 3 3 Glyceryl Monostearic Acid Aster 2 2 2 2 2  I Sedum extract 5 - glycerin 5 Na-hyluronate 5 1,3-butylene glycol 5 Purified water Quantity Quantity Quantity Quantity Quantity Quantity

Note) Unit: Weight%

division Example 14 Comparative Example 3 Comparative Example 4 Comparative Example 5 Comparative Example 6 Moisturizer Types Sedum extract - glycerin Na-hyluronate 1,3-butylene glycol Conemeter (△ Hydration)  18  5  11  7  10

Example 15 and Comparative Example 7

This Example prepared a cosmetic containing the dolmul extract obtained in Example 1 to evaluate the skin whitening effect in humans by performing a comparative experiment with Comparative Example 7.

The cosmetics used in the comparative experiments are in the form of a cream, the composition of which is shown in Table 16. First, the phase b) recorded in Table 16 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 15, Comparative Example 7). Twenty test subjects (20-35 year old female) were applied to the cream prepared in Example 15 on the right side of the face and the cream prepared in Comparative Example 7 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 17 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 17, it can be seen that the whitening effect is excellent in the facial skin of the experimenter who applied the cream containing sedum extract.

Raw material Example 15 Comparative Example 7       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 Sedum 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 15 Comparative Example 7 Example 15 Comparative Example 7 Example 15 Comparative Example 7 medium 3.89 1.47 3.2 1.9 2.5 1.3

Other examples are shown below. That is, lotion, milky lotion and essence containing the dolmul extract obtained in Example 1 was prepared in Examples 16 to 18. 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 16: Preparation of a lotion containing sedum 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 sedum 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 17 Preparation of Latex Containing the Sedum 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 dolmul 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 18 Preparation of Cosmetic Liquid Containing the Sedum 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 dolmul extract obtained in Example 1 and a suitable amount of the pigment was mixed to obtain a cosmetic liquid having a skin improvement effect.

As described above, the sedum extract according to the present invention has 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 effects to relieve skin irritation caused by UV irradiation. There is.

In addition, the sedum extract exhibits a whitening effect through melanin production inhibitory effect, which is a substance causing blemishes, freckles and skin pigmentation.

In addition, the dolmulsu extract shows an antibacterial activity and 5 alpha-reductase inhibitory effect of acne bacteria having excellent hair loss prevention effect and acne prevention effect.

Therefore, cosmetic compositions such as cosmetics, creams, emulsions, packs, and powders containing these dolmul extracts have antioxidant and collagen degrading activity control effects, skin fine wrinkles improvement effect, whitening, hair growth, acne prevention, skin irritation effect, and moisturizing. Efficacy as an excellent skin external preparation such as effects.

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)

A composition for external application for skin containing sedum extract of the genus Cirsium genus as the main active ingredient. The method of claim 1, A skin external composition comprising dolna extract as a main active ingredient, characterized in that it contains 0.001 to 30.0% by weight, based on the total weight of the composition. The method of claim 1, The sedum extract is extracted by immersion by extraction at room temperature of 70 ~ 85 ℃ using an extracting solvent, filtration of the resulting suspension obtained by further concentrating or freeze drying the liquid obtained at 50 ℃ or less Skin external composition containing the main 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. An external preparation composition for skin containing sedum extract as a main active ingredient. The method according to any one of claims 1 to 4, The skin external preparation composition is a skin external preparation composition containing as an active ingredient dolnamul extract, characterized in that for acne prevention, skin aging prevention. The method according to any one of claims 1 to 4, The skin external preparation composition is a skin external preparation composition containing as a main active ingredient dolnamul extract, characterized in that it is for inhibiting skin damage caused by ultraviolet rays or to alleviate skin irritation, whitening. The method according to any one of claims 1 to 4, The skin external preparation composition is a hair external preparation composition containing as an active ingredient dolnamul extract, characterized in that the hair damage prevention, hair growth. The method according to any one of claims 1 to 4, 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 The main activity of the sedum extract is any one selected from the cosmetic formulations consisting of base, foundation, skin cover, lipstick, lip gloss, face powder, two-way cake, eye shadow, cheek color and eyebrow pencils. Skin external composition containing as a component.