KR20150071967A - Composition for improving of skin conditions, or anti-oxidation comprising Chrysanthemum burbankii extract - Google Patents

Abstract

The present invention relates to an antioxidant skin-condition improving composition including a shasta daisy (Chrysanthemum burbankii) extract as an active ingredient. According to the present invention, the shasta daisy extract included in the composition as the active ingredient can improve the skin conditions significantly such as alleviating hypomelanosis, protecting the skin from UV rays, and preventing or alleviating inflammatory skin diseases. The shasta daisy extract also has an antioxidant activity. The present invention does not have cell toxicity and side effects to be safely applied as cosmetic and pharmaceutical compositions.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for improving skin condition or an antioxidative composition containing a chestnut extract,

The present invention relates to a composition for improving skin condition or an antioxidative composition, and more particularly, to a composition having excellent product stability, safely used without adverse effects on the skin, a skin hypochromic improving effect, a skin protecting effect from ultraviolet rays, A composition for improving skin condition and a composition for antioxidation, which comprises an effective ingredient of Shista daisy extract which is excellent in improving effect and antioxidant effect.

Recently, skin is stimulated by retinol and lactic acid which are widely used as raw materials for cosmetics as well as various pollutants such as environmental pollution and automobile exhaust gas, and skin diseases such as aging, skin inflammation such as atopic or acne, have. Specifically, the phenomenon of hormone imbalance is intensified due to various pollutants, and retinol and lactic acid, which are effective for decreasing the ability of the skin fibroblast to synthesize or reducing the immune function, and improving wrinkles and regrowth, are decomposed by ultraviolet rays, Resulting in skin damage due to ultraviolet rays.

The main cause of skin diseases such as skin damage and aging is caused by inflammation reaction in the body. Nitrogen monoxide (NO), a kind of reactive radical species in the body inflammatory reaction, is caused by immune cells such as macrophages And is known to play an important role in various physiological and pathological processes. Nitric oxide is a very unstable compound that is produced through the activation of iNOS (inducible nitric oxide synthase), a nitric oxide (NO) producing enzyme by inflammatory agents such as lipopolysaccaride (LPS). COX-1 and COX-2 are known as COX (cyclooxygenase), which is also known as PGHS (prostaglandin H synthase), and is an important regulatory enzyme that acts to convert membrane phospholipids into prostalandins (thromboxanes) . COX-1 is basically present in tissues and constantly exhibits a certain action, whereas COX-2 is expressed in inflammatory reaction sites in animals or humans, and is known to be rapidly expressed in a short period of time by various stimuli. It is known that increased expression of COX-2, a typical inflammatory enzyme, also affects the production of nitrogen monoxide (NO) (Daewoo, Journal of the Korean Society of Cosmetics (2006) 58: 173-180).

It is also known that various types of cytokines or chemokines are involved in the process of infiltrating inflammatory cells into the lesion, and related receptors are expressed in various cells such as inflammatory cells and keratinocytes. Chemokine is a family of cytokines, now known as more than 50 species. They play a role in attracting appropriate immune cells to the site while the tissue is causing an inflammatory reaction. Chemokines play an important role in the infiltration of inflammatory cells into the skin, especially in atopic dermatitis. RANTES, regulated before activation, normal T expressed and secreted, monocyte chemotactic protein 4 (MCP4) , Eotaxin-1 is frequently elevated in atopic dermatitis lesions, which is closely related to the chemotaxis of eosinophils with CCR3 receptors and Th2 cells. Eosinophils secrete highly toxic protein and free radicals that can kill microbes and parasites, but on the other hand they can be very tightly regulated as they can cause serious tissue damage to the host if they are inadequately activated Loses. These eosinophils are Eotaxin-1, which mediates the transfer of circulating blood to tissues, and Eotaxin-1 modulation is a new target for atopic treatment.

In order to improve or prevent skin damage and aging as described above, there has been conventionally used a non-steroidal antioxidant such as benzodamine, intromethasin, or ibuprofen to prevent the aging of cells by inhibiting the generation of active oxygen or dexamethasone, hydrocortisone, Using the same steroid-based antioxidants, it improved skin wrinkles and skin inflammation diseases. In addition, skin whitening and skin hyperpigmentation can be prevented by using a substance having an inhibitory activity against diroscinase such as hydroquione, ascorbic acid, kojic acid, glutathione, Improvement. However, since the above-mentioned chemicals cause side effects when they are used excessively and have a problem in stability and safety, it is necessary to develop a composition which is safe from side effects derived from natural materials and is excellent in improving skin diseases.

The skin color of a person is determined by the concentration and distribution of melanin in the skin. The melanin pigment, which is produced in melanocytes of human skin, is a phenolic high molecular substance having a complex form of black pigment and protein and plays an important role in blocking skin damage caused by ultraviolet rays. Tyrosinase, which is present in melanocytes, has been reported to be the most important for melanin biosynthesis. Tyrosinase is a type of amino acid tyrosine, which is an intermediate product of melanin polymer production (DOPA) And dopaquinone, thereby playing a key role in the skin blackening process. Melanin protects the skin from ultraviolet rays by absorbing and refracting photons from UV or IR. Other functions include body temperature control, skin protection, vitamin D synthesis, and free radical scavenging. Hypopigmented diseases caused by melanocytes being destroyed or unable to function by external stress or autoimmune disease include vitiligo, decolorized white rot, pseudoleucoderma atopicum, tineaversicolor, scleroderma, (Bolognia and Pawelek, J Am Acad Dermatol (1988) 19: 217-255; Pinto and Bolognia, Pediatr Clin North Am. (1988) 1991) 38: 991-1017). The currently reported hypoxia treatment methods include photochemical methods using photosensitizers such as psoralen, surgical methods such as surgery, methods for increasing melanin production by increasing melanin cell activity, and methods for increasing melanin cells from oxidative stress And how to protect them.

On the other hand, Daisy (English daisy, Bellis perennis ) is a perennial plant of the dicotyledonous plant, and is traditionally used as a medicinal plant. The daisies currently on the market include Shady Daisy, Dalberg Daisy, Glory Osa Daisy, Kaif Daisy, African Daisy, English Daisy, and usually Daisy refers to English Daisy. The leaves of Daisy are used for trauma or bruises, and the juice that is rooted heals eczema. In addition, it is used for liver diseases, bronchial diseases and constipation, and is excellent in detoxification and convergence, and is used as a raw material of astringent in cosmetics. In addition, it has antifungal properties and is sprayed to chase insects with juice from leaves.

In this regard, Korean Patent Registration No. 1114141 discloses a cosmetic composition having excellent skin texture improvement and elasticity-enhancing effect including daisy, marigold, safflower, geranium and pansy flower extracts, and Korea Patent No. 1145089 Japanese Laid-Open Patent Application No. 2012-201649 discloses a skin bleaching agent formulation comprising an enzymatic hydrolyzate of plum and a daisy flower extract, and a skin external preparation containing the same, wherein the skin bleaching agent formulation comprises Bellis perennis Lt; / RTI > However, there has not yet been disclosed the improvement of skin hypochromatosis, protection of skin from ultraviolet rays, improvement of inflammatory skin disease, or antioxidant efficacy using the same Shusta daisy extract as in the present invention.

Accordingly, the present inventors have made efforts to develop a plant-derived skin condition improving composition that is harmless to human body and has excellent safety and effect by using natural products. As a result, it has been found that a composition comprising an effective ingredient of Shusa daisy extract, , Skin protection effect from ultraviolet ray, inflammatory disease improving effect and antioxidant effect, and completed the present invention.

It is an object of the present invention to provide a cosmetic composition for improving skin condition comprising an extract of Shista daisies as an active ingredient.

Another object of the present invention is to provide a cosmetic composition for antioxidation comprising an extract of Shista daisies as an active ingredient.

It is still another object of the present invention to provide a pharmaceutical composition for improving skin condition comprising an extract of Shista daisies as an active ingredient.

It is another object of the present invention to provide a pharmaceutical composition for antioxidation comprising an extract of Shista daisies as an active ingredient.

The present invention relates to a cosmetic composition for skin condition improvement or antioxidation comprising an extract of Shista daisies as an active ingredient.

In the present invention, the above-mentioned Chrysanthemum burkankii is a breeding species produced by a breeder of the United States, Luther Burbank, in 1920, crossing the French marine C. leucanthemum with an oriental island chrysanthemum, , And the efficacy against the skin aging of sharda daisies, improvement of skin hypochromatosis, protection of skin from ultraviolet rays, improvement of inflammatory skin diseases, antioxidant effect and the like are not yet known.

In the present invention, the shasta daisy extract may be extracted from any one or more selected from the group consisting of leaves, roots, stems, and flowers including the outbreak of Shasta daisies, preferably flowers.

In the present invention, the Shasta daisy extract can be extracted with water or an organic solvent, and the extracted liquid can be used in a liquid form or can be used by concentration and / or drying. The organic solvent may include, but is not limited to, methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF) (DMSO), 1,3-butylene glycol, propylene glycol or a mixed solvent thereof, preferably methanol, ethanol, butanol, hexane, dichloromethane, ethyl acetate or a mixed solvent thereof, Can be extracted at room temperature or warming under non-destructive or minimized conditions. Depending on the organic solvent to be extracted, the degree of extraction and the degree of loss of the active ingredient of the extract may differ. Therefore, an appropriate organic solvent should be selected and used. The extraction method of the organic solvent is not particularly limited, and examples thereof include cold extraction, ultrasonic extraction, and reflux cooling extraction.

The purified water is a process for removing suspended solid particles from an extract or a fraction. The purified water may be filtered using a cotton, nylon, or the like, or an ultrafiltration method, a freezing filtration method, a centrifugation method, or the like. In addition, it may further comprise a separation process by various chromatographies (size, charge, hydrophobicity or affinity chromatographic).

The method of concentration and / or drying includes, but is not limited to, freeze drying, vacuum drying, hot air drying, spray drying, vacuum drying, foam drying, high frequency drying, infrared drying and the like. In some cases, a step of pulverizing the final dried extract may be added.

In the present invention, the skin condition improvement refers to improvement of skin hypochloremia, skin protection from ultraviolet rays, or prevention or improvement of inflammatory skin disease.

Therefore, the cosmetic composition of the present invention has a use for improving skin hypocholality. As used herein, the term " hypochromatosis improvement "means an action to improve skin troubles such as vitiligo and white fever caused by abnormal melanin cell or melanin pigment synthesis function and reduced melanin production.

In one specific embodiment, the Shasta daisy extract exhibits an effect of increasing the melanin production of melanocytes derived from human body, has high stability, has little change in ingredient content over time, and has little side effects such as skin irritation.

In addition, the cosmetic composition of the present invention has a skin protecting function against ultraviolet rays. The composition of the present invention protects skin cells from ultraviolet rays while lowering cytotoxicity, and more particularly, to protect melanocytes having a function of protecting the skin from ultraviolet rays, or to suppress the cell death of cells damaged by ultraviolet rays, And such facts are clearly described in the examples.

The skin protection effect from ultraviolet rays may include conventional skin protection uses (skin soothing effect, skin blackening inducing effect, skin cancer prevention effect, and the like).

In addition, the cosmetic composition of the present invention has a use for preventing or improving inflammatory skin diseases. As one example, the Shasta daisy extract of the present invention reduces the secretion of COX-2, the production-inducing enzyme of progladin, an inflammatory substance, and the production of nitrogen monoxide (NO), and the production of eotaxin- Reducing production. It also inhibits the growth of Staphylococcus aureus , which causes dermatitis.

Accordingly, the cosmetic composition of the present invention can be applied to various inflammatory diseases, diseases or abnormal conditions which can be prevented or treated by controlling the inflammatory reaction. The inflammatory disease can be, but is not limited to, for example, acne, atopic dermatitis, psoriasis, seborrheic dermatitis, contact dermatitis, irritable lupus or premature urticaria.

In the present invention, the cosmetic composition has an excellent antioxidative effect because it has an excellent free radical scavenging effect by the effective ingredient Shusa daisy extract.

The antioxidant cosmetic composition of the present invention can be applied to various antioxidative-related diseases, diseases or abnormal conditions which can be prevented or treated by inhibiting or eliminating the oxidation state. Antioxidant-related diseases that can be prevented or treated by the compositions of the present invention include, but are not limited to, atherosclerosis, coronary artery disease, coronary artery restenosis, reperfusion injury, neurodegenerative diseases such as Parkinson's or Alzheimer's disease, stroke, , Cardiovascular disease, osteoporosis, central nervous system disorder, peripheral vascular disease, and dyspnea.

As used herein, the term "prophylactic " means inhibiting the occurrence of a disease or disorder in an individual who has never been diagnosed as having a disease or condition, but is likely to be susceptible to such disease or disease. As used herein, the term " treatment "or" improvement "means (a) inhibiting the development of a disease or disorder in an individual; (b) relieving the disease or disorder; As used herein, the term " individual " means a mammal, including a human, having a disease whose symptoms may be ameliorated by administration of the composition of the present invention, such as a monkey, cow, horse, pig, sheep, dog, cat, rat, Means a mammal.

In the present invention, the cosmetic composition may contain ingredients commonly used in cosmetic compositions in addition to the above-mentioned Shusta daisy extract. For example, conventional auxiliaries acceptable as cosmetics such as antioxidants, stabilizers, solubilizers, vitamins, pigments and perfumes, and carriers.

The cosmetic composition may be formulated into a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing oil, powder foundation, emulsion foundation, wax foundation, But is not limited thereto. More specifically, it may be formulated into a nutritional cream, a convergent lotion, a soft lotion, a lotion, an essence, a shampoo, a nutritional gel or a massage cream.

Starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide as the carrier component when the cosmetic composition is a paste, cream or gel, Can be used.

When the formulation of the cosmetic composition is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In particular, in the case of spray, a mixture of chlorofluorohydrocarbons, Propane / butane or dimethyl ether.

When the cosmetic composition is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan, and the like.

When the formulation of the cosmetic composition is in the form of a suspension, the carrier component may be a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tragacanth.

When the formulation of the cosmetic composition is an interfacial active agent-containing cleansing, it is preferable to use, as carrier components, aliphatic alcohol sulfates, aliphatic alcohol ether sulfates, sulfosuccinic acid monoesters, isethionates, imidazolinium derivatives, methyltaurates, sarcosinates, fatty acids Amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

The active ingredient of the above-described Shasta daisy extract is 0.00001 to 15% by weight, preferably 0.0001 to 10% by weight, more preferably 0.0001 to 5% by weight, based on the whole composition. When the weight of the shassy daisy extract is less than 0.00001% by weight, the effect of improving skin hypochromatosis, protecting skin from ultraviolet rays, improving inflammatory disease and antioxidation is weak, and when it is more than 15% by weight, Is very weak.

The present invention also relates to a pharmaceutical composition for skin condition improvement or antioxidation comprising Shista daisy extract as an active ingredient.

In the present invention, the pharmaceutical composition may contain a pharmaceutically acceptable carrier in addition to the above-described Shusta daisy extract. The pharmaceutically acceptable carriers to be contained in the pharmaceutical composition of the present invention are those conventionally used in the present invention and include carbohydrate-based compounds such as lactose, amylose, dextrose, sucrose, sorbitol, mannitol, starch, cellulose, etc. ), Acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, salt solution, alcohol, gum arabic, vegetable oil But are not limited to, vegetable oils, soybean oil, soybean oil, olive oil, coconut oil), polyethylene glycol, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention can be administered orally or parenterally, and is preferably applied by parenteral administration, more preferably by local administration by application.

The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, . On the other hand, the dosage of the pharmaceutical composition of the present invention is preferably 0.001-100 mg / kg (body weight) per day. When the composition is an external preparation, it is preferable to apply the composition in an amount of 1.0 to 3.0 ml on an adult basis once to five times a day for one month or longer. However, the dose is not intended to limit the scope of the present invention.

The pharmaceutical composition of the present invention may be formulated into a unit dose form by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by a person having ordinary skill in the art to which the present invention belongs. Or by intrusion into a multi-dose container. The formulations may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of excipients, powders, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

The active ingredient of the above-described Shasta daisy extract is 0.00001 to 15% by weight, preferably 0.0001 to 10% by weight, more preferably 0.0001 to 5% by weight, based on the whole composition. When the weight of the shassy daisy extract is less than 0.00001% by weight, the effect of improving skin hypochromatosis, protecting skin from ultraviolet rays, improving inflammatory disease and antioxidation is weak, and when it is more than 15% by weight, Is very weak.

Meanwhile, in a specific example of the present invention, as a result of a skin irritation test on the extract of Shasta daisies, the shasta daisy extract was found to be a harmless substance as a natural substance. Therefore, since the shasta daisy extract of the present invention has little toxicity and side effects, it can be safely used even in long-term use, and can be safely applied to cosmetic and pharmaceutical compositions as described above.

The chasta daisies extract of the present invention is effective for inhibiting the growth of melanocytes, suppressing apoptosis of keratinocytes induced by ultraviolet rays, reducing nitrogen monoxide production, inhibiting COX-2 activity, decreasing Eotaxin-1 production and growth of Propionibacterium acnes It has an excellent skin improving effect. Such skin improvement includes improvement of skin hypochromatosis, skin protection from ultraviolet rays, or improvement of inflammatory disease. In addition, Shasta daisy extract has an antioxidative effect because it effectively scavenges DPPH free radicals. And can be safely applied to cosmetic and pharmaceutical compositions without cytotoxicity and skin side effects.

FIG. 1 is a graph showing the inhibitory effect of COX-2 on the expression of prostaglandin E2 according to the concentration of Shasta daisy extract. FIG.
FIG. 2 is a graph showing the effect of inhibiting the expression of Eotaxin-1 according to the concentration of Shusta daisy extract in the case where Eotaxin-2 is induced in human normal fibroblasts.
FIG. 3 is a graph showing the effect of reducing nitrogen monoxide according to the concentration of Shasta daisy extract when inducing production of nitrogen monoxide in astrocytes. FIG.
FIG. 4 is a graph showing HPPD free radical scavenging ability according to the concentration of the Shasta daisy extract. FIG.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example 1: Preparation of Shasta daisy extract

The shade daisy flower was collected and washed thoroughly to remove impurities and impurities completely, shaken at 20 to 35 캜, and pulverized to have a particle size of 1 mm or less. Then, 1 kg of the crushed Shasta daisy flower powder was immersed in 10 L of 70% methanol and sonicated for 24 hours. Then, the obtained extract was filtered through a filter paper (Advantes, No. 2) and concentrated under reduced pressure to prepare a Shasta daisy extract.

Example 2 Measurement of Melanin Production Enhancement Effect of Human Melanocytes by Shasta Daisy Extract

Melanin production by the shasta daisy extract of Example 1 was measured using human melanocytes. In this experiment, human epidermal melanocytes (from newborns) purchased from Cascade Biologics were cultured in 254 medium containing human melanocyte growth promoter. First, human epidermal melanocytes were inoculated into 6-well plates at 2 × 10 ⁵ per well and cultured at 37 ° C. under 5% CO ₂ until the cells were attached to the well-bottom at about 80% or more . After cultivation, the medium was removed, and 10, 50 and 100 ppm of Shasta daisy extract was treated and cultured for 5 days under 5% CO ₂ at 37 ° C. for ₂ days. Subsequently, the culture medium was removed from the culture medium, washed with PBS (phosphatized buffer saline), and treated with trypsin to recover the cells. The recovered cells were counted using a hematocytometer and the cells were divided into 2-mL tubes in equal numbers, centrifuged at 5,000 to 10,000 rpm for 10 minutes, and the supernatant was collected And the pellet was obtained. The pellet was dried at 60 ° C., and 100 μl of a 1M sodium hydroxide solution containing 10% DMSO was added thereto to obtain intracellular melanin in a 60 ° C. thermostatic chamber. The absorbance of this solution was measured at 490 nm using a microplate reader to determine the amount of melanin per cell constant. Melanocytes not treated with Shasta daisy extract were used as a control group. The results are shown in Table 1 below.

density Amount of melanin (% of cont) Control group (-) 100 Example 1 (10 ppm) 103 Example 1 (50 ppm) 121 Example 1 (100 ppm) 134

As a result of the experiment, the shasta daisy extract showed a concentration - dependent increase in the melanin production of human - derived melanocytes.

Example 3 Measurement of Ultraviolet Protection Effect of Shasta Daisy Extract

UV-induced keratinocytes (HaCaT) were cultured in DMEM primary culture medium consisting of penicillin-streptomycin and serum. To a 12-well microplate was added approximately 1 x 10 < RTI ID = 0.0 >⁵ Lt; RTI ID = 0.0 > 37 C < / RTI > CO₂ The cells were incubated in the incubator until approximately 80% of the cells were attached to the well bottom. 0, 1, 10 and 100 [mu] M shasta daisies extract were added 24 hours before UV irradiation. The culture medium was changed to PBS, and ultraviolet rays (UVB 20 mJ / cm²) Were investigated. After irradiation with ultraviolet light, the cells were changed to DMEM basic culture medium containing penicillin-streptomycin and serum, and cell viability was measured using MTT assay (ATCCTM, 30-1010K) at the end of culture for 48 hours. As the negative control group, keratinocytes not treated with UVB treatment and shasta daisy extract were used, and the results are shown in Table 2 as a positive control group.

Cell viability (% of cont) Non-UVB irradiated group Negative control (-) 100 UVB irradiation group (20 mJ / cm ² ) Positive control (-) 51 Example 1 (10 ppm) 60 Example 1 (50 ppm) 67 Example 1 (100 ppm) 74

The results showed that the extract of Shasta daisy inhibited the cell death of keratinocytes induced by ultraviolet light, and inhibited the cell death induced by ultraviolet irradiation as the concentration of Shuster daisy extract increased.

Example 3: Inhibitory effect of chasta daisy extract on COX-2 activity

In general, we examined the effect of Shasta daisy extract on the enzyme activity of COX-2, an enzyme that produces inflammation-inducing prostaglandins. Human monocytic cells (THP-1) were treated with 200 ng / ml of LPS (Lipopolysaccharide) and cultured at 10 and 100 ppm of Shasta daisy extract for 24 hours. After 24 hours, the cell culture medium was collected and the amount of prostaglandin E2 (PGE2) was determined by measurement. As a control (-), monocyte cells without shasta daisy extract treatment were used. The results are shown in Fig.

The results showed that Shasta daisy extract decreased the secretion of COX-2, and COX-2 secretion decreased as the concentration of Shasta daisy extract increased, indicating a concentration-dependent tendency.

Example 4: Measurement of Eosin-1 secretion inhibitory effect of Shasta daisy extract

Was inoculated so that the per well of about 2 × 10 ⁵ cells of the human normal fibroblast cells (normal human fibroblast, HNF) the 24-well microplates containing the DMEM medium, 5% CO ₂ concentration Lt; RTI ID = 0.0 > 37 C < / RTI > for 24 hours. Then, 10 and 50 ppm of Shasta daisy extract were treated respectively, and 50 ng / ml of interleukin-4 (R & D, Minneapolis MN, IL-4) was treated together for 24 hours and cell culture liquid was collected. The secretion level of eotaxin-1 was then measured using an ELISA kit. Shasta daisy extract and human fibroblasts which did not induce the secretion of eotaxin-1 were used as the control (-). The results are shown in Fig.

As a result of the experiment, it was shown that the secretion of eotaxin-1 induced by interleukin-4 was reduced by the shasta davi extract, and the secretion of eotaxin-1 was decreased with increasing the concentration of shasta daisy extract. .

Example 5 Reduction of Nitric Oxide Production of Shasta Daisy Extract

Mouse macrophages (Raw 264.7 cells, Korean Cell Line Bank) were inoculated into a 24-well microplate containing DMEM medium to a concentration of about 2 × 10 ⁵ cells per well and cultured in a 5% CO ₂ Lt; RTI ID = 0.0 > 37 C < / RTI > for 24 hours. Lipopolysaccharide (LPS) (200 ng / ㎖), which is known to induce nitric oxide synthases (iNOS), which is responsible for nitric oxide production, is then treated with 10 and 100 ppm of Shasta daisy extract and incubated for 48 hours. (1% w / v) sulfanilamide, 0.1% (w / c) N- [1-naphthyl] ethylene diamine dihydrochloride) and the same amount of a grease reagent (Griess reagent: Minute, and reacted at 540 nm. As control (-), macrophages without shasta daisy extract were used. The results are shown in Fig.

The results showed that the production of nitrogen monoxide (LPS) - induced nitric oxide was reduced by Shasta daisy extract, and as the concentration of Shasta daisy extract increased, the concentration of nitrogen monoxide decreased, indicating a concentration - dependent tendency.

Example 6: Determination of DPPH free radical scavenging effect of Shasta daisy extract

Diphenyl-2-picryl-hydrazyl (DPPH) method in order to confirm the effect of free radical scavenging of the extract of Shasta daisies. First, a 0.1 mM DPPH solution diluted in 4 ml of ethanol and 1, 10 and 100 mg / ml of Shasta daisy extract were mixed and allowed to stand at room temperature for 30 minutes, and the absorbance was measured at 517 nm. As a control group, ascorbic acid used as a conventional antioxidant was used. The results are shown in Fig.

The results showed that Shasta daisy extract significantly eradicated DPPH free radicals.

Example 7: Staphylococcus aureus of Shasta daisy extract ( Staphylococcus aureus ) On the antimicrobial activity

In order to activate Staphylococcus aureus , the cells were cultured in a 3 ml BHI liquid medium (Brain heart infusion broth; 3.7%) for 48 hours in an anaerobic incubator at 37 ° C, (Brain-Heart infusion broth; 3.7%; agar 1.5%), and dried. Then, 100 μg / ml of shasta daisies extract was dispensed on a 10 mm paper disk, and the suspension was placed on the solid medium and cultured in an anaerobic incubator at 37 ° C. for 24 hours to confirm the formation of inhibition zone. The two-fold serial dilution method was used to measure the MIC of the sharda daisy extract. Here, MIC is defined as the minimum concentration that inhibits the growth of microorganisms. The results are shown in Table 3.

Example 1 MIC concentration 0.75 mg / ml

As a result of the experiment, it was confirmed that the MIC concentration of the Shasta daisy extract on Propionibacterium acnes was 0.75 mg / ml.

Example 8: Confirmation of the safety of the extract of Sharda daisy on human skin

8-1. Manufacture of external preparations for skin including shade daisy extract

In order to confirm that the extract of Shasta daisies is safe for the human skin, the skin stability test was carried out after preparing the external preparation for skin containing the Shasta daisy extract by the ingredients and contents in Table 4 below. First, purified water, glycerin, and butylene glycol were mixed and dissolved at 70 캜 (water part), and the other components except for the above three components and trimethanolamine were dissolved at 70 캜 (oil part). The oil part was added to a water part and stirred with a homomixer (Tokushu Kika, Japan) for primary emulsification, and trimethanolamine was finally added thereto. Next, the bubbles generated in the mixed solution were removed, and the mixture was cooled to room temperature to prepare an external preparation for skin.

ingredient content (%) Control group Test group 1 Test group 2 Purified water 72 72 72 glycerin 8.0 8.0 8.0 Butylene glycol 4.0 4.0 4.0 Shasta daisy extract 0 0.1 1.0 Caprylic Capri Triglyceride 8.0 8.0 8.0 Squalane 5.0 5.0 5.0 Cetearyl glucide 1.5 1.5 1.5 Sorbitan stearate 0.4 0.4 0.4 Cetearyl alcohol 1.0 1.0 1.0 Trimethanolamine 0.1 0.1 0.1 Gross weight 100 100 100

8-2. Experiment of cumulative stimulation of skin

The skin external preparation prepared by the method of 8-1 was applied to 30 healthy adults for 9 times and 24 hours cumulatively over the upper forearm every day to determine whether or not the extract of Shasta Daisy extract stimulates the skin . As a control group, a skin external preparation which did not contain a shusa daisy extract was used as a squalane base.

A pin chamber (Finn chamber, Epitest Ltd, Finland) was used as a coating method, and 15ul of each skin external agent was dropped into the chamber, followed by applying a patch. The degree of the skin reaction on each occasion was scored using Equation 1 below, and the results are shown in Table 5 below.

[Experimental Equation 1]

The number of tests (9 times) and the number of tests (9 times)

At this time, a score of ± is given as 1, a score of 2 is given as +, and a score of 4 is given as ++, and a safe composition is judged when the average degree of reaction is less than 3.

Test substance Number of subjects who responded (persons) Average
Reactivity 1 week 2 weeks 3 weeks Primary Secondary Third Fourth 5th 6th Seventh 8th car 9th ± / + / ++ ± / + / ++ ± / + / ++ ± / + / ++ ± / + / ++ ± / + / ++ ± / + / ++ ± / + / ++ ± / + / ++ Control group 2/0/0 - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - 0.18 Test group 1 0/-/- 0/-/- - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - 0.00 Test group 2 1/0/0 0/-/- - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - 0.09 Subject 30 30 30 30 30 30 30 30 30

As a result, the numbers of ±, +, and ++ in the control group were 2, 0, and 0 in the primary pellet, respectively, and no skin reaction occurred after the secondary pellet. In test group 2, the numbers of ±, +, and ++ corresponded to 1, 0, and 0, respectively, in the primary pellet, and no skin reaction occurred after the secondary pellet. The average reactivity of the control group was 0.18, and the average reactivity of the test groups 1 and 2 was 0.00 and 0.09, respectively, which was less than 3, which was judged as a safe composition.

Formulation Example 1: Formulation of cosmetic preparation

1-1. Production of flexible lotion

As shown in Table 6 below, a lotion lotion containing an extract of Shasta daisies as an active ingredient was prepared according to a conventional method.

ingredient Content (% by weight) Shasta daisy extract 0.01 glycerin 3.0 Butylene glycol 2.0 Propylene glycol 2.0 Carboxyvinyl polymer 0.1 ethanol 10.0 Triethanolamine 0.1 Preservatives, micronutrients, trace fragrances and trace water Balance sum 100.0

1-2. Manufacture of nutrition lotion

As shown in Table 7 below, a nutritional lotion containing an extract of Shasta daisies as an active ingredient was prepared according to a conventional method.

ingredient Content (% by weight) Shasta daisy extract 0.01 Wax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 Liquid paraffin 5.0 Squalane 5.0 Caprylic / capric triglyceride 5.0 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxyvinyl polymer 0.1 Triethanolamine 0.2 Preservatives, micronutrients, trace fragrances and trace water Balance sum 100.0

1-3. Manufacture of nutrition cream

As shown in Table 8 below, a nutritional cream containing an extract of Shistar daisy as an active ingredient was prepared according to a conventional method.

ingredient Content (% by weight) Shasta daisy extract 0.01 Wax 10.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 Liquid paraffin 10.0 Squalane 5.0 Caprylic / capric triglyceride 5.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservatives, micronutrients, trace fragrances and trace water Balance sum 100.0

1-4. Manufacture of massage cream

As shown in Table 9 below, a massage cream containing the chasta daisies extract as an active ingredient was prepared by a conventional method.

ingredient Content (% by weight) Shasta daisy extract 0.01 Wax 10.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.8 Liquid paraffin 40.0 Squalane 5.0 Caprylic / capric triglyceride 4.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservatives, micronutrients, trace fragrances and trace water Balance sum 100.0

1-5. Manufacture of packs

As shown in the following Table 10, packs containing the Shasta daisy extract as an active ingredient were prepared according to a conventional method.

ingredient Content (% by weight) Shasta daisy extract 0.01 Polyvinyl alcohol 13.0 Sodium carboxymethylcellulose 0.2 Allantoin 0.1 ethanol 5.0 Nonyl phenyl ether 0.3 Preservatives, micronutrients, trace fragrances and trace water Balance sum 100.0

Preparation Example 2: Preparation of pharmaceutical preparations

2-1. Manufacture of Powder

In order to prepare powders containing an extract of Shistar daisy as an active ingredient, the ingredients shown in the following Table 11 were mixed and filled in airtight bags to prepare powders.

ingredient Content (g) Shasta daisy extract 2 Lactose One

2-2. Manufacture of tablets

In order to prepare tablets containing the shistada daisy extract as an active ingredient, tablets were prepared by mixing the ingredients shown in the following Table 12 and then tableting according to a conventional method for producing tablets.

ingredient Content (mg) Shasta daisy extract 100 Corn starch 100 Lactose 100 Magnesium stearate 2

2-3. Preparation of capsules

In order to prepare capsules containing an extract of Shistar Daisy as an active ingredient, the following ingredients of Table 13 were mixed and filled in gelatin capsules according to the conventional preparation method of capsules to prepare capsules.

ingredient Content (mg) Shasta daisy extract 100 Corn starch 100 Lactose 100 Magnesium stearate 2

Claims (10)

A cosmetic composition for improving skin condition comprising an extract of Chrysanthemum burbankii as an active ingredient, wherein the improvement of skin condition is characterized by improvement of skin hypochlorite , protection of skin from ultraviolet rays, prevention or improvement of inflammatory skin disease Cosmetic composition.
The cosmetic composition according to claim 1, wherein the skin hypochloremia is vitiligo or white moth.
The cosmetic composition according to claim 1, wherein the inflammatory skin disease is at least one of acne, atopic dermatitis, psoriasis, seborrheic dermatitis, contact dermatitis, irritable lupus, or acute urticaria.
An antioxidant cosmetic composition comprising an extract of Chrysanthemum burbankii as an active ingredient.
5. The cosmetic composition according to any one of claims 1 to 4, wherein the Shasta daisy extract is extracted from at least one selected from the group consisting of leaves, roots, stems and flowers including Shasta daisies.
A pharmaceutical composition for improving skin condition comprising Chrysanthemum burbankii extract as an active ingredient, wherein the improvement of skin condition includes prevention of skin aging, prevention and improvement of hypochlorite skin, protection of skin from ultraviolet rays or prevention of inflammatory skin disease And an improvement.
7. The pharmaceutical composition according to claim 6, wherein the skin hypochromatosis is vitiligo or white moth.
[Claim 7] The pharmaceutical composition according to claim 6, wherein the inflammatory skin disease is at least one of acne, atopic dermatitis, psoriasis, seborrheic dermatitis, contact dermatitis, erythrocyte lupus, or acute urticaria.
An antioxidant pharmaceutical composition comprising an extract of Chrysanthemum burbankii as an active ingredient.
[Claim 10] The pharmaceutical composition according to claim 6 or 9, wherein the Shasta daisy extract is extracted from at least one selected from the group consisting of leaves, roots, stems and flowers including Shasta daisies.

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