KR102468821B1 - Cosmetic Composition for Blue Light Interception - Google Patents

Abstract

The present invention relates to a cosmetic composition for blocking blue light containing a plant complex extract, and more particularly, to a cosmetic composition comprising nasturtium extract, kale leaf extract, dandelion leaf extract, turnip leaf extract, carrot sprout extract, pumpkin extract, and sweet potato root extract, watermelon extract, and turmeric rhizome extract containing at least one of blue light blocking as an active ingredient, as well as skin cell protection and skin regeneration, antioxidant, suppression or prevention of skin inflammatory response, skin wrinkles It relates to a cosmetic composition characterized in that it is for improving, preventing or suppressing, enhancing skin activity, regulating skin temperature, and soothing skin and a method for manufacturing the same.

Description

Cosmetic composition for blocking blue light {Cosmetic Composition for Blue Light Interception}

The present invention relates to a cosmetic composition for blocking blue light, and more particularly, to a cosmetic composition for blocking blue light containing a plant complex extract.

Blue light is a type of visible light that emits a short wavelength that exists between 380 and 500 nanometers, and refers to blue-based light. It is emitted a lot from the displays of smart devices such as TVs, computers, and mobile phones, and from LED lighting devices, so that objects can be seen clearly and people feel comfortable. However, long-term exposure to blue light in everyday life causes eye fatigue as well as dry eye syndrome, and in severe cases, it may cause damage to the retina or lens in the eye. In addition, blue light late at night inhibits the secretion of sleep-inducing hormones, causing errors in the circadian rhythm, which interferes with sound sleep and makes it difficult to regenerate damaged skin during the day.

Among ultraviolet rays, which is a type of visible light, UVB penetrates down to the epidermis, and UVA penetrates into the dermis layer, but blue light penetrates into the subcutaneous tissue layer deeper than the dermis layer, so it can be more harmful to the skin. Because of this intense energy, it is also called HEV (High-Energy Visible) light. Therefore, when directly exposed to the skin for a long period of time, it promotes melanin production and causes pigmentation such as age spots and melasma, as well as increased active oxygen, resulting in oxidative stress, resulting in premature aging symptoms such as inflammation, wrinkles, and loss of elasticity of the skin. can

Recently, in order to prevent skin damage and acceleration of aging due to excessive exposure of blue light, research on harmful mechanisms of the skin is in progress. Research on blue light blocking materials and related formulations in the field of cosmetics is still in its infancy, and test methods related to blue light blocking measurements have not been clearly established. Safflower seed oil (Prior Document 1), phytoplankton extract (Prior Document 2), and the like are known as cosmetic materials for blocking blue light.

Prior Document 1: Korean Patent Application Publication 10-2019-0006910 Prior Document 2: Korean Registered Patent No. 10-2031289

As a result of diligent efforts to discover materials having functionality for blocking blue light, the inventors of the present invention found nasturtium extract, kale leaf extract, dandelion leaf extract, turnip leaf extract, carrot sprout extract, pumpkin extract, sweet potato root extract, and watermelon extract. , And it was found that any one or more of the turmeric rhizome extracts had this effect, and the present invention was completed.

Therefore, an object of the present invention is to use any one or more of nasturtium extract, kale leaf extract, dandelion leaf extract, turnip leaf extract, carrot outpost extract, pumpkin extract, sweet potato root extract, watermelon extract, and turmeric rhizome extract as active ingredients. It is to provide a cosmetic composition for blocking blue light containing a.

Another object of the present invention is to use any one or more of the active ingredients of nasturtium extract, kale leaf extract, dandelion leaf extract, turnip leaf extract, carrot seedling extract, pumpkin extract, sweet potato root extract, watermelon extract, and turmeric rhizome extract. It is to provide a method for producing a cosmetic composition for blocking blue light containing a.

In order to solve the above problems, the present invention is a nasturtium extract, kale leaf extract, western dandelion leaf extract, turnip leaf extract, carrot outpost extract, pumpkin extract, sweet potato root extract, watermelon extract, and turmeric rhizome extract any one or more It provides a cosmetic composition for blocking blue light containing as an active ingredient.

Hereinafter, the present invention will be described in more detail.

In one aspect, the present invention is an active ingredient of any one or more of nasturtium extract, kale leaf extract, dandelion leaf extract, turnip leaf extract, carrot seedling extract, pumpkin extract, sweet potato root extract, watermelon extract, and turmeric rhizome extract. It relates to a cosmetic composition for blocking blue light containing. Specifically, the extract is not limited in its content, but may be contained in an amount of 0.1 to 10% by weight based on the total weight of the composition.

In the present invention, a description of each plant of each plant cell serving as an active ingredient is as follows.

1. Tropaeolum Majus Extract: Nasturtium is native to Mexico and South America, and is derived from the Greek word 'tropaion', which means round shield-like leaves and helmet-like flowers. Flowers bloom in June, and one stalk comes out from the axil, and one flower hangs at the end. The flower color is red, orange, yellow, etc., and there is also a mancheop flower. In Europe, it is called victory flower and is planted in pots and flower beds. The flowers emit flashes of light in mid-summer because they contain high concentrations of phosphoric acid. The nasturtium is mainly used for ornamental purposes or as a fragrance, and contains a large amount of iron and vitamin C. Its leaves, flowers and fruits have tonic, blood purifying and disinfecting effects. Leaves or flowers are applied to bruises or dermatitis, and the leaves are decocted to treat bronchitis or genitourinary infections.

2. Kale Leaf Extract (Brassica Oleracea Acephala Leaf Extract): Kale, which is native to the Mediterranean, has various nutrients and is used as a representative super food. It is a vegetable with the highest beta-carotene content among green and yellow vegetables, and is also very rich in lutein, which is good for eye health. In addition, kale is one of the foods that can consume the most vitamin K, which helps blood clotting and bone health, and is also rich in calcium and magnesium, which is good for preventing osteoporosis. The short one is 30 ~ 60 cm, the tall one is about 120 cm, and it grows well in any soil with sufficient moisture. The leaves grow wide, long and thick like tobacco leaves, and receive plenty of sunlight to turn dark green.

3. Western dandelion leaf extract (Taraxacum Officinale (Dandelion) Leaf Extract): Native to Europe, it is a naturalized plant that came to Korea about 100 years ago and is common in each region. It is a plant. The taste of 'Tarashqun' in Persian is derived from the meaning of a bitter, boiled vegetable. In Europe, it is a plant that has been widely used as a medicine because it not only consumes leaves as a salad and roots as a coffee substitute, but also contains a large amount of functional ingredients such as vitamins and minerals. Flowers are yellow in February-March and bloom in multiple single flowers. Seeds are long oval with hairs attached, and these seeds gather to become fluffy fruits like fluffy hair.

4. Brassica Rapa (Turnip) Leaf Extract: Originated from southern Europe along the Mediterranean coast, turnips with a water content of more than 90% are root vegetables that contain many nutrients in their roots and leaves. It is rich in vitamins and contains a lot of chlorophyll, beta-carotene, and carotenoid pigments, helping to prevent cancer and aging. The leaves of turnips contain many minerals and vitamins, but the roots contain a lot of tryptophan and lysine. The size and shape of the root also differs depending on the variety, but it is usually a top-shaped round shape. The difference between a radish and a radish is the color of the root, which is purple, unlike a radish. The taste is spicy but savory and has a mustard-scented ginseng taste. It is also called ‘fruit radish’ because it tastes sweet unlike ordinary radish.

5. Carrot Outpost Extract (Daucus Carota Sativa (Carrot) Extract): Carrots are native to the Himalayas, to which Afghanistan belongs, and are also called carrots and are about 1m tall. Carrots with a cone-shaped shape and a scarlet color are known as good eyesight protection foods because of their high content of vitamin A among vegetables. Rich in beta-carotene, lutein, and lycopene, it has an antioxidant effect, helps prevent aging, improves immunity, and prevents cancer, high blood pressure, and arteriosclerosis. In Korea, it can be cultivated all year round, so it can be easily met at any time, and it is widely used for its unique scent and color.

6. Western pumpkin extract (Cucurbita Maxima Fruit Extract): Western pumpkin, also called zucchini pumpkin or pork pumpkin, is native to the Americas and is the most popular summer pumpkin in the Americas and Europe. It is yellow, green, or light green, and looks very similar to zucchini and cucumbers, and some cultivars grow in the shape of a round gourd. Most zucchini have a soft skin, small edible seeds, and crisp, soft flesh with lots of moisture. As one of the very low-calorie vegetables, the skin contains a lot of dietary fiber that reduces constipation and prevents colon cancer. In addition, it is rich in polyphenolic antioxidants such as carotene, lutein and zeaxanthin. It is also an excellent source of potassium, one of the important intracellular electrolytes. Potassium is a good electrolyte for the heart and counteracts the pressure effect of sodium, lowering blood pressure and heart rate.

7. Sweet potato root extract (Ipomoea Batatas Root Extract): The origin of sweet potato is tropical America. As a vine plant that spreads along the ground and takes root, sweet potatoes grow in size as nutrients are accumulated in the root, and the part we eat represents this root. This kind of root is called a tuberous root, and the sweet potato is purplish brown on the outside and white on the inside and has a dense structure. Sweet potatoes are especially rich in flavonoid phenolic compounds such as beta-carotene and vitamin A among root vegetables. It has excellent effects on skin beauty, so if you take one a day, you can have healthy skin. In addition, rich vitamin E is a powerful antioxidant that protects cells from free radicals, thereby slowing down skin aging.

8. Citrullus Lanatus (Watermelon) Fruit Extract: Watermelon is native to Africa and has been cultivated since the time of ancient Egypt. It is a fruit of a plant belonging to the Cucurbitaceae family. It has a spherical or oval shape with a dark green rind and weighs about 3 to 5 kg. The pink flesh has a slight difference in light and shade, and is very refreshing and has a little sweetness, but there is no special taste. The flesh is flat and has black seeds embedded in it. It is a typical fruit eaten in summer. It has a very high water content and low calories, so it releases a lot of water and contains a large amount of lycopene, which has excellent antioxidant effects. In addition, it contains an amino acid called citrulline, so it has a high diuretic effect and is said to be good for nephritis.

9. Curcuma Longa (Turmeric) Rhizome Extract: Turmeric is native to tropical Asia and belongs to the genus Ginger and Curcuma. The history is long enough that there is a story that it was brought as an emergency medicine when needed. If turmeric, a curry ingredient, is dried rhizome of turmeric, turmeric is dried tuberous root of turmeric. Turmeric is often referred to as a ‘God-sent food ingredient’ because it contains antioxidant flavonoids, calcium, potassium, and various other vitamins and minerals. In particular, curcumin, the main ingredient, is a component that makes turmeric yellow, and is effective in preventing cardiovascular diseases such as diabetes and high blood pressure and dementia. It has excellent anti-inflammatory and anti-cancer effects that inhibit inflammation or tumor growth, and recently it is not only a great help for skin beauty, but also used for anti-cancer treatment. It is also known to help improve liver function as it has the effect of expelling bad substances from the body out of the body.

In the present invention, the composition for blocking blue light, in addition to blocking blue light, for this reason or separately, for skin cell protection and skin regeneration, for antioxidant, for suppressing or preventing inflammatory reactions in the skin, for improving skin wrinkles, for prevention Or, it includes various skin activity improvement functions such as suppression, skin activity enhancement, and skin soothing.

The manufacturing method of the cosmetic composition for blocking blue light according to the present invention includes the following steps:

(a) preparing any one or more of nasturtium extract, kale leaf extract, dandelion leaf extract, turnip leaf extract, carrot seedling extract, pumpkin extract, sweet potato root extract, watermelon extract, and turmeric rhizome extract; and (b) preparing a composition containing the extract prepared in step (a).

In the present invention, step (b) may include preparing a composition containing each extract obtained in step (a).

In the present invention, in step (a), an extract can be prepared by mixing nasturtium, kale leaf, dandelion leaf, turnip leaf, carrot whole plant, pumpkin, sweet potato root, watermelon and turmeric root.

For example, each ratio is 14 to 17 parts by weight (g): 54 to 66 parts by weight (g): 14 to 17 parts by weight (g): 54 to 66 parts by weight (g): 54 to 66 parts by weight ( g): 9 to 11 parts by weight (g): 9 to 11 parts by weight (g): 54 to 66 parts by weight (g): 3 to 5 parts by weight (g).

In the present invention, step (a) may include preparing an extract by ultrasonic extraction or hot water extraction.

In the present invention, "extract" refers to an extract obtained by various conventionally known extraction methods such as cold water extraction, hot water extraction, and ethanol extraction of plant tissues. When preparing such an extract, it is possible to use the tissues of the various plants presented above, and its parts are also not limited to leaves, roots, stems, fruits, or outposts.

In the present invention, the extraction method is not particularly limited, and examples thereof include cold needle extraction, ultrasonic extraction, reflux extraction, and hot water extraction. Here, in the case of hot water extraction, hot water extract is obtained by heating at 80 to 100 ° C. for 8 to 48 hours in a hot water distillation machine.

In the case of cold water extraction, for example, the plant tissue itself or its dried powder is mixed with cold water (15-25° C.) and extracted for 3 days to obtain a cold water extract.

Alternatively, it may be prepared by a method of extraction using water, an organic solvent, or a mixed solvent thereof. The extracted liquid may be used directly or may be used after being concentrated and/or dried. When using an organic solvent for extraction, methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixture of these organic solvents are used, and the active ingredients of herbal medicines can be extracted at room temperature or heated under conditions that are not destroyed or minimized. Depending on the organic solvent to be extracted, the degree of extraction and loss of the active ingredient of the drug may differ, so select and use an appropriate organic solvent.

In the present invention, the extract may be used after being concentrated or diluted, and a distillate of the extract may be used.

In the present invention, the meaning of "contained as an active ingredient" is a cosmetic composition for blocking blue light, as well as for skin cell protection and skin regeneration, in relation to or separately from it, for antioxidant, and for suppressing or preventing inflammatory reactions of the skin. It means that it contains an effective amount to the extent that it can exhibit functionalities such as skin wrinkle improvement, prevention or suppression, skin activity enhancement, and skin soothing.

In the cosmetic composition, a carrier acceptable for cosmetic formulations may be included. Here, "acceptable carrier in cosmetic preparation" means a compound or composition that is already known and used, or a compound or composition to be developed in the future, which can be included in a cosmetic preparation, and has toxicity, instability or irritation more than the human body can adapt to when in contact with the skin. say nothing

The carrier may be included in the cosmetic composition of the present invention in an amount of about 1% to about 99.99% by weight, preferably about 90% to about 99.99% by weight, based on the total weight of the composition. However, since the ratio varies depending on the formulation as described below in which the cosmetic composition of the present invention is prepared and its specific application area (face, neck, etc.) or its preferred application amount, the ratio is the present invention in any aspect should not be construed as limiting the scope of

Alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, humectants, viscosity modifiers, emulsions, stabilizers, UV scattering agents, UV absorbers, coloring agents, perfumes, and the like may be exemplified as the carrier. Compounds/compositions that can be used as the alcohol, oil, surfactant, fatty acid, silicone oil, wetting agent, humectant, viscosity modifier, emulsion, stabilizer, UV scattering agent, UV absorber, coloring agent, fragrance, etc. are already known in the art. Since there is, those skilled in the art can select and use an appropriate corresponding material / composition.

As an embodiment of the present invention, the cosmetic composition according to the present invention may include glycerin, butylene glycol, propylene glycol, polyoxyethylene hydrogenated castor oil, ethanol, triethanolamine, etc. in addition to the extract as the active ingredient, preservative , fragrances, coloring agents, purified water, etc. may be included in small amounts as needed.

The cosmetic composition according to the present invention can be prepared in various forms, for example, lotion, essence, gel, emulsion, lotion, cream (oil-in-water type, water-in-oil type, multi-phase), solution, suspension (anhydrous and aqueous) , anhydrous products (oil and glycol-based), gels, masks, packs, powders, or capsules (soft capsules, hard capsules) with a coating such as gelatin.

Skin in the present invention is a concept that includes not only the face, but also the scalp and the whole body, and as a cosmetic composition that can be applied to such a scalp, there are shampoo, rinse, treatment, hair growth agent, etc., body cleanser that can be applied to the whole body, etc. It can be manufactured in various forms for use.

The manufacturing method of the cosmetic composition according to the present invention is not limited to the above-mentioned manufacturing method, and those skilled in the art may also manufacture a method partially modified from the above manufacturing method.

When prepared as a cosmetic composition, emulsified cosmetics include nutritional lotion, cream, essence, etc., and solubilized cosmetics include softening lotion. In addition, by containing a dermatologically acceptable medium or base, it can be prepared in the form of an adjuvant that can be applied topically or systemically, which is commonly used in the field of dermatology.

In addition, suitable cosmetic formulations include, for example, solutions, gels, solid or pasty anhydrous products, emulsions obtained by dispersing an oil phase in an aqueous phase, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes), nonionic It may be provided in the form of a vesicular dispersion of the type, cream, toner, lotion, powder, ointment, spray or conceal stick. In addition, it can be prepared in the form of a foam (foam) or the form of an aerosol composition further containing a compressed propellant.

In addition, the cosmetic composition of the present invention may further include a fatty substance, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, water, and an ionic type. or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or any other commonly used in cosmetics. It may contain adjuvants commonly used in the field of cosmetology or dermatology, such as ingredients. In addition, the above ingredients may be introduced in an amount generally used in the field of dermatology.

The cosmetic composition for blocking blue light according to the present invention is a natural material that can effectively block blue light, and has various functions such as anti-inflammatory, skin regeneration, skin soothing, skin heat control, and skin wrinkle improvement.

Figure 1 shows the results of skin cell culture of the composition according to the present invention.
Figure 2 is the result of confirming the skin cell death protection and regeneration effect from blue light of the composition according to the present invention.
3 is a result of confirming whether or not the CAT activity of the composition according to the present invention is increased from blue light.
Figure 4 shows the results of confirming the anti-inflammatory effect of the composition according to the present invention.
Figure 5 shows the results of confirming the cell damage protection of the composition according to the present invention.
Figure 6 is the result of confirming the anti-wrinkle effect of the composition according to the present invention.
Figure 7 is the result of confirming the cell energy activity enhancing effect of the composition according to the present invention.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is one well known and commonly used in the art. The percentages mentioned in the examples below are understood as volume % or weight %.

Example 1: Preparation of a cosmetic composition for blocking blue light according to the present invention

16 g nasturtium thoroughly washed in 20 L of purified water, 60 g kale leaves, 16 g dandelion leaves, 60 g turnip leaves, 60 g carrot seedlings, 10 g zucchini, 10 g sweet potato roots, 60 g watermelon, 4 g turmeric The rhizome was put in and hot water extraction was performed while heating at 80 to 100 ° C. for about 8 to 48 hours. After extraction, the solid content was removed by filtration through a mesh, and a composite extract was obtained.

Experimental example One: skin cells Test to confirm regeneration effect by non-stimulation and cell proliferation through culture

In order to determine whether the composition of the present invention causes toxicity in skin cells and causes skin irritation, MTT assay was performed. HaCaT cells (human keratinocytes, keratinocytes) and Detroit cells (human fibroblasts, fibroblasts, Detroit 551) were plated in a 96-well plate with DMEM (Dubelcco's Modified Eagle's Medium) medium containing 10% FBS (Fetal Bovine Serum). It was divided and cultured for 24 hours in an incubator under conditions of 5% CO ₂ and 37°C. Thereafter, the cells were treated with the control group and the composition of the present invention to which purified water was added to a final concentration of 0.1%, 1%, 5%, 10%, and 20%, and further cultured for 24 hours. Subsequently, 4 μL of 5 mg/mL MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) solution was added to each well and incubated for 4 hours. After removing the culture medium, 100 μL of DMSO (Dimethyl sulfoxide) solution was added to dissolve the cells, and the absorbance was measured at 540 nm using a spectrophotometer.

As a result, as shown in FIG. 1, it was confirmed that the composition of the present invention did not show toxicity in skin cells at a treatment concentration of 0.1 to 20% and thus did not cause irritation. Therefore, the composition means that the possibility of causing skin irritation is low and it is a safe material.

Experimental Example 2: Skin cell protection and regeneration effect confirmation test

In order to determine whether the composition according to the present invention is involved in protecting and regenerating skin cells from blue light irradiation, MTT assay was performed. To this end, HaCaT cells and Detroit cells were dispensed into a 96-well plate with DMEM (Dubelcco's Modified Eagle's Medium) medium containing 10% FBS (Fetal Bovine Serum) and incubated in a thermostat under 5% CO ₂ and 37 °C for 24 hours. cultured. After that, the cells were treated with the control group and the test material composition to which purified water was added to a final concentration of 0.1, 1, 5, and 10%, and after 2 hours, Topview 5450 VRI SMD LED (obtained from: ITSWELL, Part no. IWS-L5056 -VRI-K3) was used to irradiate blue light (114 mW/cm ² , 30 minutes). Cells were then further cultured for 24 hours. Subsequently, 4 μL of 5 mg/mL MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) solution was added to each well and incubated for 4 hours. After removing the culture medium, 100 μL of DMSO (Dimethyl sulfoxide) solution was added to dissolve the cells, and the absorbance was measured at 540 nm using a spectrophotometer.

As a result, as shown in Figure 2, it was confirmed that the composition of the present invention protects the death of skin cells from blue light and has a regenerative effect at a treatment concentration of 0.1 to 10%.

Experimental example 3: blue light Blocking activity evaluation

In order to evaluate the blue light blocking effect of the composition of the present invention, the blue light wavelength band (380 to 500 nm) absorbance of each sample was measured. The control group and the composition of the present invention were diluted to a final concentration of 1% in purified water, and the spectrum of 380 to 500 nm was analyzed with a spectrophotometer.

Composition of the present invention (%) Blue light absorption rate (%) One 0 41.8 2 0.1 51.4 3 One 85.8 4 5 91.7 5 10 88.2 6 20 61.2

As a result, as shown in Table 1, as the concentration of the composition according to the present invention increased to 5%, the blue light absorption rate increased significantly compared to the control group, and at a concentration of 5% or more, it was confirmed that the blue light absorption rate gradually decreased. did The higher the blue light absorption rate, the higher the blue light blocking effect.

Experimental example 4: Antioxidant effect confirmation test by CAT activity

In order to examine the antioxidant activity of the composition of the present invention, the activity of CAT (Catalase) known to be involved in the decomposition of H ₂ O ₂ or reactive oxygen species (ROS), which are toxic substances generally generated in aerobic metabolism CAT assay was performed to investigate.

To this end, HaCaT cells were seeded in a 12-well plate and cultured for 24 hours under cell culture conditions. After that, the cells were treated with the control group and the test material composition to which purified water was added to a final concentration of 0.1, 1, 5, and 10%, and after 2 hours, Topview 5450 VRI SMD LED (obtained from: ITSWELL, Part no. IWS-L5056 -VRI-K3) was used to irradiate blue light (114 mW/cm ² , 30 minutes). Cells were then further cultured for 24 hours. Take 25 μL of the diluted lysate of the cells cultured through the above process and put it in a 96-well plate together with 25 μL of the CAT standard material in the CAT assay kit (Dogenbio, Cat. DG-CAT400), respectively, and add 40 μM H ₂ O ₂ 25 μL of the solution was also added. After reacting at 37 ℃ for 3 hours, 50 μL of the Oxi-Probe/HRP working solution was added to each well in which the reaction was completed. Thereafter, the mixture was reacted at 37° C. for 30 minutes in a shaded state, and absorbance was measured at 560 nm. After measuring the absorbance at 560 nm, a CAT standard concentration curve was prepared to calculate the amount of CAT in the sample.

As a result, as shown in Figure 3, it was confirmed that the composition of the present invention has an antioxidant effect by increasing the activity of CAT from blue light at a treatment concentration of 0.1 to 10%.

Experimental example 5: Test for confirming the inhibitory effect of inflammatory gene activation

In order to examine the skin inflammation inhibitory effect of the composition of the present invention, changes in the gene expression levels of COX-2 (Cyclooxygenase-2) and iNOS (Inducible nitric oxide synthase) involved in inflammatory reactions were investigated.

To this end, HaCaT cells were seeded in a 96-well plate and cultured for 24 hours under cell culture conditions. After that, the cells were treated with the control group and the test material composition to which purified water was added to a final concentration of 0.1, 1, 5, and 10%, and after 2 hours, Topview 5450 VRI SMD LED (obtained from: ITSWELL, Part no. IWS-L5056 -VRI-K3) was used to irradiate blue light (114 mW/cm ² , 30 minutes). Cells were then further cultured for 24 hours. Real-time PCR was performed to confirm COX-2 and iNOS at the gene level, and the test sequence is as follows.

For RNA isolation and cDNA synthesis, SuperPrep ^™ cell lysis & RT kit for qPCR (TOYOBO, Cat. SCQ-101) was used. After removing the medium, the cells were washed once with PBS, 50 μL cell lysis mixture (including gDNA remover) was added, reacted for 5 minutes, and then stop solution was added. 8 μL of the extracted mRNA was added to 32 μL of the RT reaction mixture, and cDNA was synthesized using PCR at 37°C for 15 minutes, 50°C for 5 minutes, and 95°C for 5 minutes. To compare and analyze gene expression, the cDNA synthesized above was used as a template and real-time PCR analysis was performed using Thunderbird ^™ SYBR qPCR Mix (TOYOBO, Cat. QPS-201). The primers used in the experiment were Qiagen's QuantiTect primer assays (GAPDH; Cat. QT01192646, COX-2; Cat. QT00040586, iNOS; Cat. QT01323189), and the mRNA expression levels of COX-2 and iNOS in the samples were quantified and compared with GADPH. . Real-time qPCR conditions were first performed at 95 ° C for 15 minutes, followed by 94 ° C for 15 seconds, 60 ° C for 30 seconds, and 72 ° C for 30 seconds per cycle, for a total of 40 cycles.

As a result, as shown in FIG. 4, it was confirmed that the composition of the present invention has an anti-inflammatory effect by reducing the expression rate of COX-2 and iNOS, which are inflammatory response related genes, from blue light at a treatment concentration of 0.1 to 10%.

Experimental example 6: antistress Protein activation effect confirmation test

The activity level of LEA (Late embryogenesis abundant) protein was investigated to determine whether the composition of the present invention has a protective effect against intracellular blue light stress. LEA is known to protect cells from damage by increasing resistance to stress as the expression level increases.

To this end, HaCaT cells were seeded in a 6-well plate and cultured for 24 hours under cell culture conditions. After that, the cells were treated with the control group and the test material composition to which purified water was added to a final concentration of 0.1, 1, 5, and 10%, and after 2 hours, Topview 5450 VRI SMD LED (obtained from: ITSWELL, Part no. IWS-L5056 -VRI-K3) was used to irradiate blue light (114 mW/cm ² , 30 minutes). Then, the cells were additionally cultured for 24 hours, and an In-Cell ELISA Colorimetric Detection kit was used to measure the amount of LEA protein expression.

The medium of the cultured cells was removed, and the cells were fixed by culturing at room temperature for 15 minutes with 100 μL fixing solution. After washing with PBS, 100 μL of Permeabilization solution was added and reacted at room temperature for 15 minutes. After washing with PBS, 100 μL quenching solution was added and reacted at room temperature for 20 minutes. After washing with PBS, 100 μL of blocking solution was added and reacted at room temperature for 30 minutes. After removing the solution, 50 μL of the LEA primary antibody was added and reacted at 4° C. for 24 hours. After removing the antibody, it was washed with PBS, and 50 μL of HRP conjugated secondary antibody was added and reacted for 1 hour. After removing the antibody and washing with PBS, 100 μL of TMB substrate solution was added and reacted at room temperature for 15 minutes in a shaded state. Then, 100 μL stop solution (1 NH ₂ SO ₄ ) was added, and absorbance was measured at 450 nm.

As a result, as shown in FIG. 5, it was confirmed that the composition of the present invention protects cells from damage by increasing the expression level of LEA protein from blue light at 0.1 to 10% treatment concentration.

Experimental example 7: by collagen enhancement anti-wrinkle effect confirmation test

A PIP assay was conducted to determine whether the composition of the present invention enhances the amount of collagen production related to wrinkle improvement.

To this end, Detroit cells were seeded in a 48-well plate and cultured for 24 hours under cell culture conditions. After that, the cells were treated with the control group and the test material composition to which purified water was added to a final concentration of 0.1, 1, 5, and 10%, and after 2 hours, Topview 5450 VRI SMD LED (obtained from: ITSWELL, Part no. IWS-L5056 -VRI-K3) was used to irradiate blue light (114 mW/cm ² , 30 minutes). Cells were then further cultured for 24 hours. Take 20 μL of the diluted supernatant of the medium cultured through the above process, put it into an antibody coated microtiter plate together with 20 μL of the PIP standard in the PIP EIA kit (Takara, Cat. MK101), and 100 μL of peroxidase-labeled antibody solution was also added to each well and reacted at 37 °C for 3 hours. After removing the medium and washing 4 times with 200 μL of PBS, 100 μL of substrate solution was added to each well and reacted at room temperature for 15 minutes in a shaded state. Then, 100 μL stop solution (1 NH ₂ SO ₄ ) was added, and absorbance was measured at 450 nm. After measuring absorbance at 450 nm, a PIP standard concentration curve was prepared to calculate the amount of PIP in the sample.

As a result, as shown in FIG. 6, it was confirmed that the composition of the present invention increased the amount of PIP expression from blue light at a treatment concentration of 0.1 to 10%, resulting in collagen proliferation.

Experimental example 8: Cell energy enhancement effect test

In order to examine the cell energy enhancing effect of the composition of the present invention, the activity level of ATP released by cells during metabolism was measured.

To this end, Detroit cells were seeded in a 12-well plate and cultured for 24 hours under cell culture conditions. Thereafter, the cells were treated with the composition as a test substance and the control group to which purified water was added to a final concentration of 0.1, 1, 5, and 10%, and after 2 hours, blue light was irradiated using a Topview 5450 VRI SMD LED (114 mW/cm ² , 30 minutes). Cells were then further cultured for 24 hours. Take 50 μL of the diluted lysate of the cells cultured through the above process and put it in a 96-well plate together with 50 μL of the ATP standard in the ATP assay kit (Abcam, Cat. ab83355), respectively, and 50 μL of ATP reaction mix solution was also added to each well and reacted at room temperature for 30 minutes in a shaded state. After measuring the absorbance at 570 nm, an ATP standard concentration curve was prepared to calculate the amount of ATP in the sample.

As a result, as shown in Figure 7, it was confirmed that the composition of the present invention has an energy activity enhancing effect by increasing the amount of ATP from blue light at a treatment concentration of 0.1 to 10%.

Experimental example 9: Evaluation of skin soothing effect (skin temperature reduction rate)

The effect of preventing heat aging by the composition of the present invention was evaluated. That is, after formulating a lotion to contain the composition according to the present invention by concentration, it was applied to the skin to confirm the effect of preventing heat aging. The test was performed on the forearm of 25 healthy subjects in their 20s and 30s, and the test site (forearm) of the subject was exposed to direct sunlight for 20 minutes before the test. The test sample and control sample were applied in an amount of 50 μL each using a micro pipette to the selected test area (2 cm * 2 cm) of the forearm, and the temperature before, immediately after, 5 minutes after, and 20 minutes after application of the sample was measured. Water was used as a negative control for comparison of heat aging prevention efficacy, and the temperature reduction rate of each sample after measurement was derived according to the following equation.

Heat reduction rate (%) = (1 - temperature of sampled area / temperature of non-sampled area) * 100

Heat reduction rate (%) Composition of the present invention (%) immediately after application 5 minutes later after 20 minutes 0 6.1 2.1 0 0.1 7.0 3.5 0.8 0.5 9.2 4.5 1.5 One 17.3 8.4 2.8 5 18.1 8.8 2.9 10 19.5 10.4 4.1

As a result, in the case of the control group, the temperature decrease rate rapidly decreased over time, but it was confirmed that the sample containing the composition of the present invention maintained a constant heat decrease rate after 20 minutes in a concentration-dependent manner even after 20 minutes.

Claims (9)

A cosmetic composition for blocking blue light containing nasturtium extract, kale leaf extract, dandelion leaf extract, turnip leaf extract, carrot seedling extract, pumpkin extract, sweet potato root extract, watermelon extract, and turmeric rhizome extract as active ingredients. The cosmetic composition according to claim 1, wherein the composition is for skin cell protection and skin regeneration. The cosmetic composition according to claim 1, wherein the composition is for antioxidation. The cosmetic composition according to claim 1, wherein the composition is for suppressing or preventing an inflammatory reaction of the skin. The cosmetic composition according to claim 1, wherein the composition is for improving, preventing or inhibiting skin wrinkles. The cosmetic composition according to claim 1, wherein the composition is for enhancing skin activity. delete The cosmetic composition according to claim 1, wherein the composition is for soothing the skin. (a) preparing nasturtium extract, kale leaf extract, dandelion leaf extract, turnip leaf extract, carrot spring extract, pumpkin extract, sweet potato root extract, watermelon extract, and turmeric rhizome extract; and
(b) mixing the extract prepared in step (a) with purified water and heating to obtain a composite extract after hot water extraction,
Claims 1 to 6 and 8 containing nasturtium extract, kale leaf extract, dandelion leaf extract, turnip leaf extract, carrot starch extract, pumpkin extract, sweet potato root extract, watermelon extract, and turmeric rhizome extract. Method for producing a cosmetic composition according to any one of the preceding.

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