KR20130099610A - Skin external composition containing sceptridium ternatum extract - Google Patents

Abstract

PURPOSE: An external use skin composition containing a Sceptridium ternatum (Thunb.) Lyon extract is provided to improve whitening, anti-aging, and moisturizing effects. CONSTITUTION: An external use skin composition contains 0.001-50 wt% of a Sceptridium ternatum (Thunb.) Lyon extract as an active ingredient. The composition is used for whitening, anti-aging, elasticity, anti-wrinkling, and moisturizing effects.

Description

Skin external composition containing fern extract {Skin external composition containing Sceptridium ternatum extract}

The present invention is a fern ( Sceptridium) The present invention relates to an external composition for skin containing ternatum ( Thunb .) Lyon ) extract, and more particularly, to an external composition for skin that can provide a whitening effect, an anti-aging effect, and an improved skin moisturizing effect by containing a fern extract. .

Human skin is changed by a number of internal and external factors as it ages. That is, internally, the secretion of various hormones that regulate metabolism decreases, and the function of immune cells and the activity of cells decreases, thereby reducing the biosynthesis of immune proteins and constituent proteins necessary for living organisms. Due to the increase in the amount of ultraviolet rays that reach the surface of the sun's rays and intensifying environmental pollution, free radicals and free radicals increase, thereby reducing the thickness of the skin, increasing wrinkles, reducing elasticity, The color of the skin becomes dull, skin problems frequently occur, and there are various changes such as blemishes, freckles, and black mushrooms.

As aging progresses, symptoms of alteration or decrease in the content and arrangement of collagen, elastin, hyaluronic acid and glycoproteins, which are constituents of the skin, appear and are subject to oxidative stress caused by free radicals and active harmful oxygen. In addition, cyclooxygenase-2 (Cox-2, cyclooxygenase), an enzyme that produces proinflammatory cytokine, which is known to cause inflammation in most cells constituting the skin due to aging or ultraviolet rays. Biosynthesis of the enzyme increases the biosynthesis of matrix metalloproteinase (MMP), an enzyme that breaks down skin tissues by these inflammatory factors, and the production of nitric oxide (NO) by iNOS (inducible nitric oxide synthase). It is known to increase. In other words, the biosynthesis of the substrate material is reduced by the reduction of cellular activity and micro-inflammation due to naturally occurring endogenous aging, and by external factors such as the increase of stress caused by various harmful environments and the increase of active coral species caused by sunlight. As a result, decomposition and degeneration are accelerated, and the skin matrix is destroyed and thinned, and various symptoms of skin aging appear. Therefore, many studies have been conducted on active ingredients that can prevent and improve these aging phenomena.

Many factors are involved in determining the skin color of humans, including the activity of melanocytes that make melanin, the distribution of blood vessels, the thickness of the skin and the presence or absence of pigments in the body, such as carotenoids and bilirubin. It is important.

Especially, the most important factor is the black pigment called melanin which is produced by the action of various enzymes such as tyrosinase in melanocytes in the human body. The formation of melanin pigment affects genetic factors, hormonal secretion, physiological factors such as stress, and environmental factors such as ultraviolet irradiation.

The melanin pigment produced in melanocytes of body skin is a phenolic polymer substance having a complex form of black pigment and protein. It protects the subcutaneous skin organs by blocking ultraviolet rays irradiated from the sun, and at the same time, free radicals And protects proteins and genes in the skin.

Melanin, which is produced by stress stimuli inside and outside the skin, is a stable substance that does not disappear until the skin is excreted through skin keratinization even if the stress disappears. However, when melanin is produced more than necessary, it induces hypercholesterolemia such as spots, freckles, and dots, resulting in poor cosmetic results.

Today, Asian women prefer white and clean skin like white jade, and this is an important standard of beauty, increasing the demand for treatment and cosmetic problems for hyperpigmentation.

In response to this demand, ascorbic acid, kojic acid, arbutin, hydroquinone, glutathione or derivatives thereof, or substances having tyrosinase inhibitory activity have been used in combination with cosmetics or pharmaceuticals, but they are insufficient. Its use has been limited due to its whitening effect, safety issues on the skin, formulation and stability problems when formulated in cosmetics, and the like.

The most important function of the outermost epidermis of the skin is the defense against various external stimuli (physical and chemical stimulating factors such as chemicals, air pollutants, dry environment, ultraviolet rays) and moisture in the body through the skin. It is a protective function to prevent excessive divergence of, and this protective function can maintain its function only when the stratum corneum composed of keratinocytes is normally formed. The outermost stratim corneum (horney layer) of the epidermis is formed from keratinocytes and consists of the differentiated keratinocytes and the surrounding lipid layer (J. Invest. Dermatol. 1983; 80: 44 ~ 44). 49). Keratinocytes are characteristic cells whose basal cells, which continuously proliferate in the epidermal layer, undergo morphological and functional changes in stages and rise to the surface of the skin. Exfoliated and new keratinocytes take over the function. This repetitive process of change is called 'epidermal differentiation' or 'keratinization'. Keratinocytes during keratinization form the stratum corneum by generating natural moisturizing factor (NMF) and intercellular fat (ceramide, cholesterol, fatty acids), making the stratum corneum firm and flexible, making it a skin barrier. It has a function as).

However, the stratum corneum can easily lose its function due to excessive washing, lifestyle factors such as bathing, environmental factors such as dry air and pollutants, and endogenous diseases such as atopic or aged skin. In recent years, more and more people are complaining of dry skin symptoms and all kinds of obstacles due to various factors. Therefore, in order to maintain proper skin moisture, studies have been conducted to supply moisture from the outside or to prevent water loss from the body, and a variety of moisturizers having moisture retention ability have been developed, It is widely used in the area.

However, as the risk factors for humans increase in the living environment and the aging population rapidly increases, the turnover rate of the stratum corneum is delayed, the lipid synthesis ability of the keratinocytes decreases, The cells are not smoothly divided, maturated and differentiated, and the amount of moisturizing factors and lipids in the stratum corneum is decreased, so that the normal stratum corneum does not maintain its function. Namely, .

Due to the abnormal division and differentiation of epidermal cells, the skin causes various skin diseases such as dry skin, atopy and psoriasis, and these diseases can alleviate the symptoms slightly with a conventional moisturizer having only water retention function. However, it is difficult to expect radical healing.

Accordingly, the present inventors have found that fern ginseng extract from natural extracts can provide a whitening, anti-aging, skin wrinkle improvement and moisturizing improvement effect, and completed the present invention.

Accordingly, it is an object of the present invention to provide an external composition for skin that contains fern extract, exhibits the effects of whitening, anti-aging and the like and can improve moisturizing properties.

In order to achieve the above object, the present invention provides an external composition for skin containing fern extract as an active ingredient.

The present invention also provides a whitening, anti-aging and moisturizing skin external preparation composition containing fern ginseng extract as an active ingredient.

The composition of the present invention can provide a skin whitening effect, anti-aging effect and skin moisturizing improvement effect by containing fern extract.

The external preparation composition for skin of the present invention is a fern ( Sceptridium) ternatum ( Thunb .) Lyon ) extract as an active ingredient.

Fern ( Sceptridium) ternatum ( Thunb .) Lyon is a perennial fern native to the high mountains of Jeju, Jirisan, Deogyusan, Gyeongnam, Gyeongbuk, Gangwon, and Gyeonggi-do. The growing environment grows in half-shape grasses with high humidity and rich soil. The height is about 50cm, and the leaves have spores without spores and spores with spores. The leaflets are thick and glossy, and petioles are divided into three branches, long, and sawtooth at the end. Spores are 5cm long, longer than trophic leaves, horn-shaped, and have spores like millet. Spore sac is formed in September to November.

Fern ginseng extract used in the present invention is not only a leaching solution obtained by leaching and transferring in bracken, but also a concentrate obtained by partially or fully condensing the leaching solution or stagnation, whole, regular, flow extract and bracken prepared by drying the concentrate again. It contains all the chemicals in the plant, as well as the main effect of the plant itself. In addition, in the present invention, extracts from all parts of bracken, such as stems, roots, and leaves, may be used, and are not limited to extracts of any particular part.

As a method for producing fern ginseng extract used in the present invention, it is possible to use a known method. For example, the bracken ginseng of the present invention is chopped and dried, and then water or an organic solvent is added, reflux extraction is carried out, and the residue and the filtrate are separated by filtration and filtration and centrifugation, and the filtrate is concentrated under reduced pressure. You can get it. The organic solvent usable in the present invention may be selected from ethanol, methanol, butanol, ether, ethyl acetate, chloroform or a mixed solvent of these organic solvents and water, and considering the safety of the raw material, preferably 100% water, 70% ethanol Or butylene glycol (Butylene Glycol) is used. At this time, the extraction temperature is preferably 18 to 80 ℃ for water, 18 to 40 ℃ is preferable for 70% ethanol and butylene glycol, the extraction time is preferably 6 to 168 hours. If the extraction temperature and the extraction time is out of the extraction efficiency may be lowered or a change of components may occur. After obtaining the extract using the solvent in the above can be obtained by cooling, heating and filtration at room temperature in a conventional manner known in the art to obtain a liquid, or may further evaporate the solvent, spray drying or freeze drying.

The external preparation composition for skin according to the present invention may contain the bracken extract in an amount of 0.001 to 50% by weight, preferably 0.1 to 10% by weight, and more preferably 0.5 to 5% by weight, based on the total weight of the composition. If the content of the fern ginseng extract is less than 0.001% by weight, the efficacy and effect of the extract is weak, and if it exceeds 50% by weight, there is a problem in skin safety or formulation.

The composition of the present invention can be used as a composition for whitening, which can provide an excellent whitening effect by inhibiting tyrosinase activity and inhibiting melanin production.

The composition of the present invention can be used as a composition for external application for skin for anti-aging, which is excellent in improving skin elasticity and improving wrinkles.

The composition of the present invention may be used as a moisturizing skin external preparation composition, which may enhance skin barrier function and induce differentiation of skin keratinocytes. Therefore, it can be usefully used as an external composition for skin to prevent or improve skin dryness, atopic dermatitis, contact dermatitis, or psoriasis caused by incomplete epidermal differentiation.

The composition according to the invention may be formulated containing a cosmetically or dermatologically acceptable medium or base. These are all formulations suitable for topical application, for example emulsions, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes) and non- It may be provided in the form of an ionic vesicle dispersant or in the form of a cream, skin, lotion, powder, ointment, spray or cone stick. It can also be used in the form of a foam or in the form of an aerosol composition further containing a compressed propellant. These compositions may be prepared according to conventional methods in the art.

In addition, the composition according to the present invention may further comprise at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickening agents, gelling agents, softening agents, antioxidants, suspending agents, stabilizing agents, Such as fillers, emulsifiers, emulsifiers, fillers, sequestering agents, chelating agents, preservatives, vitamins, blockers, moisturizers, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredient commonly used in cosmetics May contain adjuvants commonly used in the field of cosmetics or dermatology. Such adjuvants are introduced in amounts commonly used in the cosmetics or dermatological fields.

In addition, the composition of the present invention may contain a skin absorption promoting substance in order to increase the skin whitening effect, skin aging improvement effect and skin moisturizing improvement effect.

Hereinafter, the constitution and effects of the present invention will be described in more detail with reference to test examples and formulation examples. However, these test examples and formulation examples are provided for illustrative purposes only for the sake of understanding of the present invention, and the scope and scope of the present invention are not limited by the following examples.

Example 1 Preparation of Fern Ginseng Extract

The dried and dried Fern ginseng was extracted by refluxing three times for 24 hours using 70% ethanol as an extraction solvent, and then cooled and filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure and freeze dried at 45 ° C or lower.

[Test Example 1] Elastase activity inhibition assay

The inhibition of elastase activity of fern extract was measured by comparing with phosphoramidone. The elastase and substrate used were commercially purchased from Sigma-Aldrich Co., Ltd. (Cat. No. E0127), and fern extract was used in Example 1.

Elastase activity inhibitory activity was tested by the following test method.

50 μL of a 20 μg / mL elastase type III solution was mixed in a 96 well plate with 10 mg / L Tris-HCL buffer (pH 8.0) to 10 ppm of fern extract of Example 1 above. Phosphoramidone 10ug / ml was used as a positive control, and the untreated group as a negative control was using distilled water. Thereafter, 100 µL of 0.4514 mg / mL N-SUCCINYL-ALA-ALA-ALA-p-NITROANILIDE prepared with the above buffer was added and reacted at 25 ° C for 15 minutes. After completion of the reaction, the absorbance at a wavelength of 415 nm was measured. A blank test was performed in the same manner to correct.

The method for calculating the activity of inhibiting the activity of the elastase is shown in the following Equation 1, and the results are shown in Table 1 below.

A: absorbance at wavelength 415 nm in the absence of the test substance and addition of the enzyme

B: absorbance at a wavelength of 415 nm in the absence of the test substance and no enzyme addition

C: Absorbance at wavelength 415 nm in addition of test substance and enzyme addition

D: Absorbance at wavelength 415 nm in addition of test substance and enzyme-free addition

compound Suppression degree (%) Untreated group 0 Phosphoramidone 35 Example 1 38

As shown in Table 1, the degree of inhibition of elastase activity of the fern extract was significantly inhibited compared to the untreated group as a control group, and the effect of the level was higher than that of phosphoramidon known as an elastase inhibitor. It can be seen that the fernase inhibitory effect of the fern extract of the present invention is excellent.

Test Example 2 Collagenase (MMP-1) Inhibitory Activity

Inhibition of collagenase (MMP-1, substrate metalloprotease-1) production of bracken extract of the present invention was measured in comparison with retinoic acid.

Place human fibroblasts at 5,000 cells / well in 96-well microtiter plates containing DMEM (Dulbecco's Modified Eagle's Media) containing 2.5% fetal calf serum. Incubated in a CO ₂ 5%, 37 ℃ incubator until the growth of about 80%. Fern ginseng extract was treated for 24 hours at a concentration of 10 ㎍ / ml, and then the cell culture was collected. In addition, the retinoic acid treated with the same concentration for comparison was used.

The degree of collagenase production of cell cultures was measured using a commercially available collagenase measuring instrument (Amersham Pharmacia, USA, Catalog #: RPN 2610). First, the cell culture solution collected in a 96-well plate uniformly coated with primary collagenase antibody was placed in an incubator for 3 hours. After 3 hours, the chromophore-bound secondary collagen antibody was placed in a 96-well plate and reacted for another 15 minutes. After 15 minutes, color-causing substances (3,3 ', 5,5'-tetramethylbenzidine, sigma) were added to induce color development at room temperature for 15 minutes, and 1M sulfuric acid was added to stop the color reaction. The degree of yellow was different according to the degree of reaction progress.

The absorbance of the yellow 96-well plate was measured at 405 nm using an absorbance meter, and the degree of synthesis of collagenase was calculated by Equation 2 below, and the results are shown in Table 2 below. Indicated. At this time, the reaction absorbance of the cell culture liquid collected from the group not treated with the composition was used as a control.

compound Expression degree (%) Untreated group 100 Retinoic acid 75 Example 1 76

As shown in Table 2, it can be seen that the degree of collagenase expression of bracken extract is similar to the collagenase expression inhibitory effect compared to retinoic acid known as a collagenase expression inhibitor.

Through these results, it can be confirmed that the fern extract of the present invention has an effect of inhibiting the proteolytic protease (MMP-1).

Reference Example 1 Preparation of Formulation Example 1 and Comparative Formulation Example 1

Nutritional cream was prepared in a conventional manner according to the composition of Table 3 (unit: wt%).

Compounding ingredient Formulation Example 1 Comparative Formulation Example 1 Purified water To 100 To 100 Fern Extract One - Vegetable hydrogenated oil 1.50 1.50 Stearic acid 0.60 0.60 Glycerol stearate 1.00 1.00 Stearyl alcohol 2.00 2.00 Polyglyceryl-10 Pentastearate & Behenyl Alcohol & Sodium Stearoyl Lactylate 1.00 1.00 Arakidil Behenyl Alcohol & Arakidil Glucoside 1.00 1.00 Cetylaryl Alcohol & Cetearyl Glucoside 2.00 2.00 PEG-100 Stearate & Glycerolate & Propylene Glycol 1.50 1.50 Caprylic / Carric Triglycerides 11.00 11.00 Cyclomedicone 6.00 6.00 Preservative, incense Suitable amount Suitable amount Triethanolamine 0.1 0.1

Test Example 3 Confirmation of Skin Elasticity Improvement Efficacy

In order to confirm the effect of improving skin elasticity in humans, the formulations of Formulation Example 1 and Comparative Formulation Example 1 were evaluated as follows.

Twenty healthy women in their 30s to 40s were divided into two groups of 10 people in two groups of Formulation Example 1 and Comparative Formulation Example 1, and the nourishing cream was applied to the face once a day for 12 weeks. 575, C + K Electronic Co., Germany) was used to measure skin elasticity. The results are shown in Table 4 below. The results in Table 4 are described as ΔR8 (R8 (left) -R8 (right)) values of Cutometer SEM 575, where R8 values represent the nature of skin viscoelasticity.

Experimental product Skin elasticity effect Formulation Example 1 -0.56 Comparative Formulation Example 1 -0.10

As shown in Table 4, Formulation Example 1 containing the bracken extract of the present invention was further increased skin elasticity compared to the group to which Comparative Formula 1 was applied.

Therefore, it can be confirmed that the composition containing the fern ginseng extract of the present invention is very effective for improving skin elasticity.

[Test Example 4] Confirmation of skin wrinkle improving effect

Formulation Example 1 and Comparative Formulation Example 1 were used to confirm the effect of improving wrinkles in humans by the composition of the present invention.

In order to confirm the wrinkle improvement effect of the Formulation Example 1 and Comparative Formulation Example 1 was evaluated as follows. Twenty healthy women in their 40s were divided into two groups of 10 people for each of two groups of Formulation Example 1 and Comparative Formulation Example 1, and the nourishing cream was applied to the face once a day for 12 weeks. The state of the image was measured by a skin meter (visiometer, SV600, Courage + Khazaka electronic GmbH, Germany) and analyzed. The results are shown in Table 5 below, it can be determined that the larger the (-) value the better the wrinkle improvement effect. The values in Table 5 below are the average values obtained by subtracting the pre-application parameter values from the respective parameter values after 12 weeks of application.

8 weeks after use
Clinical results R1 R2 R3 R4 R5 Formulation Example 1 -0.30 -0.21 -0.14 -0.10 -0.09 Comparative Formulation Example 1 0.04 0.03 0.01 -0.01 0.00

R1: difference between the highest and lowest of the wrinkle contour

R2: Divide the wrinkle contours by 5 spaces, and then average the R1 values

R3: The highest value among the R1 values divided by five

R4: The mean value of the baseline of the fold contour minus the top and valley values of each angle

R5: Difference value of the baseline of the wrinkle contour line minus each wrinkle outline

As shown in Table 5, the external preparation composition of Formulation Example 1 was found to have a very good skin wrinkle improvement effect.

Test Example 5 Measurement of Increased Skin Moisturizing Effect

In order to measure the effect of fern extract on increasing skin moisturizing power, Formulation Example 1 and Comparative Formulation Example 1 were used, and evaluated as follows.

Twenty male and female adults in their 40s to 50s, classified as dry skin, were divided into two groups of 10 people for two groups, Formulation 1 and Comparative Formulation Example 1, respectively. Skin moisture meter at constant temperature and humidity conditions (24 ° C, 40% relative humidity) before start of application, after 1 week, 2 weeks, 4 weeks after application, and after 2 weeks (total 6 weeks) after application is stopped. Skin moisture content was measured by (Corneometer CM825, C + K Electronic Co., Germany). The results are shown in Table 6 below. The results in Table 6 show the percentage increase of the measured value after treatment for a period of time based on the skin moisture meter value measured immediately before the start of the test.

Test group Moisture growth rate (%) 1 week Two weeks 4 weeks 6 weeks Formulation Example 1 35 34 36 28 Comparative Formulation Example 1 30 32 32 15

In the results of Table 6, when the Comparative Formulation Example 1 was applied, it showed a water increase rate of about 32% up to 4 weeks after the application was made, but after stopping the application, the moisture content of the skin decreased, but the fern extract was contained. In the case of application of Formulation Example 1, it can be confirmed that even after stopping the application, the skin moisture increase rate of 28% or more is shown. Through this, the composition of the present invention containing fern extract can be seen that the skin moisturizing effect is excellent.

Test Example 6 Measurement of Keratinocyte Differentiation Promoting Effect

In order to examine the differentiation promoting effect of keratinocytes of fern extract, the amount of CE (Cornified Envelop) generated during the differentiation of keratinocytes was measured using absorbance.

First, the keratinocytes of the primary cultured humans isolated from the epidermis of the newborn were put in a culture flask and adhered to the bottom, and then the test substance of Table 7 was treated at 5 ppm in the culture medium. It was incubated for 5 days until it grew about%. At this time, the low calcium (0.03mM) treated group and the high calcium (1.2mM) treated group were used as negative and positive controls, respectively. Then, the cultured cells were harvested and washed with PBS (Phophate buffered saline), followed by 10 mM Tris-HCl buffer (Tris-HCl, containing 2% SDS (sodium dodecyl sulfate) and 20 mM Dithiothreitol (DTT). pH 7.4) 1 ml was added to sonication, boiling and centrifugation, and the precipitate was suspended in 1 ml of PBS again to measure absorbance at 340 nm. Separately, a portion of the solution after the sonication was taken to measure the protein content and used as a reference when evaluating the degree of cell differentiation. The results are shown in Table 7 below.

Test substance The ability to differentiate in keratinocytes (%) Low calcium (0.03 mM) solution
(Negative control) 100 High calcium (1.2 mM) solution
(Positive control group) 210 Example 1 180

As shown in Table 7, it was confirmed that the treatment of bracken extract was excellent in promoting the differentiation of keratinocytes.

Test Example 7 Measurement of Skin Barrier Function Restoration Effect

In order to measure the effect of fern extract on the recovery of damaged skin barrier function due to skin damage, the following experiment was performed. Ten adult men and women's upper and lower limbs were damaged using a tape stripping method for 7 days while applying the 16 groups of Formulation Example 2 and Comparative Formulation Example 2, each of which was prepared using the composition shown in Table 8 below. The recovery of transdermal moisture loss (TWEL) was measured once a day using a Vapometer (Delfin, Finland). Comparative Formulation Example 2 is a vehicle as the negative control group. The experimental results are shown in Table 9 below. The results in Table 9 were compared on the basis of the 100% difference before treatment and after barrier damage.

Compounding ingredient Formulation Example 2 Comparative Formulation Example 2 Purified water 69 70 Propylene glycol 30 30 Example 1 One -

Test group TWEL change (%) Before processing 1 day 2 days 3 days 4 days 5 days 6 days Formulation Example 2 100 115 103 78 50 35 14 Comparative Formulation Example 2 100 121.4 112.7 98.3 70.5 62.3 43.5

As can be seen in Table 9, it can be seen that when treated with bracken extract, the transdermal moisture loss returns to normal and barrier damage is recovered.

Test Example 8 Whitening Effect

<Tyrosinase inhibitory effect>

Tyrosinase enzyme was extracted from the mushroom (Mushroom) was used as the Sigma (SIGMA). First, the substrate tyrosine was dissolved in distilled water to make a solution of 0.3 mg / ml, and the solution was added to the test tube by 1.0 ml. Then, 1.0 ml of potassium-phosphate buffer solution (0.1 mol concentration, pH 6.8) and 0.7 ml of distilled water were added thereto. Added.

0.2 ml of the sample solution prepared by mixing the bracken extract of the present invention in an ethanol solution at an appropriate concentration was added to the reaction solution, and then reacted for 10 minutes in a 37 ° C thermostat. In this case, the control group was prepared by adding only 0.2 ml of solvent instead of each sample solution, and ascorbic acid was used as a positive control group. 0.1 ml of a tyrosinase solution of 2500 units / ml was added to the reaction solution and reacted for 10 minutes in a 37 ° C thermostat. The reaction tube containing the reaction solution was placed in iced water, quenched to stop the reaction, and the absorbance at 475 nm was measured with a photospectrometer. The results are shown in Table 10 below. Each tyrosinase inhibitory effect was calculated by the following equation.

Test substance Tyrosinase inhibition rate (%) Control group (no addition) 0 Ascorbic acid 52 Example 1 51

In the results of Table 10, the tyrosinase inhibitory rate of bracken extract used in the present invention showed a similar whitening effect to the known tyrosinase inhibitor ascorbic acid, the whitening effect was very excellent.

Inhibitory Effect of Melanin Production Using B16 / F10 Melanoma Cells

Example 1 and a sample containing 0.001% by weight of kojic acid as a comparative substance, respectively, were added as a test substance to the culture medium of B16 / F10 melanoma cells (Korea Cell Line Bank) at a constant concentration and then cultured for 3 days. Removed, washed with PBS and dissolved the cells with 1N NaOH absorbance at 405nm was measured. The cells without test substance were used as a control group, and the degree of inhibition of melanin production of each test substance was measured by comparing with the melanin content of the control group. According to Equation 4, the melanin production inhibition rate is calculated and the results are shown in Table 11.

Test substance Melanogenesis inhibition rate (%) Control group
(No additives) 0 Kojic acid 53 Example 1 49

In the results of Table 11, it can be seen that the melanin production inhibition rate of bracken extract according to the present invention is very similar to kojic acid, a known melanin production inhibitor, so that the whitening effect is very excellent.

Reference Example 2 Preparation of Formulation Example 3 and Comparative Formulation Example 3

To the lotion was prepared in a conventional manner according to the composition of Table 12 (unit: wt%).

Raw material name Formulation Example 3 Comparative Formulation Example 3 1. Cetearyl alcohol 1.0 1.0 2. Chin type glyceryl stearate 1.0 1.0 3. Glyceryl stearate SE 1.5 1.5 4. Phyto squalane 3 3 5. Hydrogenated polydecene 2 2 6. Dimethicone 0.5 0.5 7. Polysorbate 60 One One 8. Sorbitan sesquioleate 0.4 0.4 9. Methylparaben 0.1 0.1 10. Propylparaben 0.05 0.05 11. Purified water To 100 To 100 12. Butylene glycol 5 5 13. Polyacrylate-13 * Polyisobutene * Polysorbate 20 0.5 0.5 14. Fern Ginseng Extract (Example 1) One -

Test Example 9 Whitening Effect Experiment on Human Skin

12 healthy men were attached to the subject's upper arm with an opaque tape with a square hole of 1.5 cm in width and length, respectively, and then 1.5 to 2 times the UV of each subject's minimum erythema (Minimal Erythema Dose). (UVB) was irradiated to induce skin blackening.

After irradiation, the formulation of Example 3 and Comparative Example 3 was applied as a test substance, and one place was prepared with a control site where nothing was applied, and the state change was observed for 8 weeks. At 4, 6, and 8 weeks, the color of the skin was measured with a colorimeter CR2002 (Minolta, Japan). The difference between the skin color (ΔL *) at the start and completion of the application was calculated according to Equation 5 below, and the results are shown in Table 13. The whitening effect was determined by comparing ΔL * between the sample application site and the control site.

division The degree of skin color brightness (DELTA L *) Measure note 4 weeks 6 weeks 8 weeks Formulation Example 3 0.3 1.3 1.9 Comparative Formulation Example 3 0.2 0.6 1.2

As can be seen in Table 13, Formula 3 containing the bracken extract of the present invention can be confirmed that the whitening effect on the human skin is significantly higher than Comparative Formula 3 containing no bracken extract.

Claims (7)

Fern ( Sceptridium) A composition for external application for skin containing ternatum ( Thunb .) Lyon ) extract as an active ingredient. According to claim 1, wherein the fern extract is an external composition for skin composition contained in an amount of 0.001 to 50% by weight relative to the total weight of the composition. The external preparation composition for skin according to claim 1, wherein the composition is for whitening. The external preparation composition for skin according to claim 1, wherein the composition is for anti-aging. The external preparation composition for skin according to claim 1, wherein the composition is for improving skin elasticity. According to claim 1, wherein the composition is a skin external composition for improving the skin wrinkles. The external preparation composition for skin according to claim 1, wherein the composition is for moisturizing.