KR101780486B1 - Cosmetic composition comprising echinacea purpurea extract - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising an extract of Echinacea, in particular a stem, a leaf and a flower extract. Eekenia (stem, leaf, flower) extract has no cytotoxicity, inhibits various cytokines and NO activities mediated by inflammatory reaction, and a composition containing the same can be applied as a cosmetic composition having anti-inflammatory effect And the extract of Echinacea (stem, leaf, flower) according to the present invention has an excellent antioxidative effect, anti-inflammatory effect and skin barrier strengthening effect and is thus very useful as a cosmetic and dermatological external agent.

Description

COSMETIC COMPOSITION COMPRISING ECHINACEA PURPUREA EXTRACT <br> <br> <br> Patents - stay tuned to the technology COSMETIC COMPOSITION COMPRISING ECHINACEA PURPUREA EXTRACT

The present invention relates to a cosmetic composition comprising an extract of Echinacea, in particular a stem, a leaf and a flower extract. The present invention is the result of the research of the leading industrial development business of the metropolitan economy carried out with the support of the Ministry of Commerce, Industry and Energy, the Korea Industrial Technology Development Agency, and the Jeju Regional Business Evaluation Institute.

In human skin, various physicochemical changes occur in the aging process, and the causes are largely divided into intrinsic aging and photo-aging. In other words, free radicals may be activated due to ultraviolet rays, stress, disease state, environmental factors, wounds, and aging. If such conditions are deepened, the antioxidant defense network in the living body may be destroyed, Thereby promoting adult diseases and aging. More specifically, lipids, proteins, polysaccharides and nucleic acids, which are major components of the skin, are oxidized to destroy skin cells and tissues, resulting in skin aging.

Particularly, the oxidation of proteins is caused by severing collagen, hyaluronic acid, elastin, proteoglycan, fibronectin, which are connective tissues of skin, And when it gets worse, mutation of DNA, cancer induction, immune function deterioration occurs.

Therefore, free radicals or ultraviolet rays generated during the metabolism of the body, ultraviolet radiation, and free radicals mediated by the inflammatory reaction are destroyed to protect the cell membrane, and already damaged cells are regenerated by active metabolism, Should be proliferated so that the skin can quickly recover and maintain healthy skin.

Human body defense can be protected from pathogen by immune response. Biological defense mechanisms against foreign microorganisms such as viruses and bacteria are divided into innate immunity and specific immunity, (Klaus, B., Immunology Letters., 19, pp. 183-192, 1988), which is mainly secreted from cells.

Lipopolysaccaride (LPS), also known as endotoxin, is present in the extracellular membrane of Gram-negative bacteria and is known to be involved in various inflammatory factors such as TNF- (tumor necrosis factor-), IL-6 , and IL-1 (inter leukin-1) (An, SJ et al., Int. Immunopharmacol., 2, 1173 - 2000, Ashutosh, KM et al., Cell Immunol., 219, pp 1 - 10, 2002). Multifunctional cytokines such as TNF- are expressed not only in normal tissues but also in the lesion process, and play an important role in skin inflammation, especially in cancer-promoting processes (Groves, RW et al. J. Dematol., 32, pp. 345-352 1994; Wakefield, PE, et al., J. Am. Acad. Dermatol., 24, pp. 675-685, 1991). It has already been reported that TNF- is associated with inflammatory skin diseases in humans (Pigue P. F., et al., J. Exp. Med., 173, pp. 673-679, 1991).

Another strong inflammatory mediator, nitric oxide (NO), is produced from L-arginine by NO synthase (NOS), and is produced by a variety of substances, such as UV, external stress, endotoxin or cytokines Lt; / RTI &gt; cells. These inflammatory stimuli increase the expression of inducible NOS (iNOS) in the cells, thereby inducing NO production in the cells and activating the macrophages to cause an inflammatory reaction.

 Recently, studies have been conducted on substances capable of inhibiting NO production for effective inflammation relief. However, some side effects of anti-inflammatory substances developed by these studies are problematic.

When sebum components, sweat components, and fatty acids, higher alcohols, proteins, and the like in the components of cosmetics, which are discharged from the body, are decomposed into highly toxic substances by skin-derived bacteria existing on the skin, they may cause skin inflammation, It is well known that skin inflammation is caused by ultraviolet rays from the sun.

In this way, the causes of skin adverse effects in cosmetics are always present and various studies have been conducted to solve them. It is a natural substance-derived substance. It has little or no danger of side effects or cytotoxicity, and can inhibit the production of NO effectively, thus it has excellent anti-inflammatory effect and can inhibit inflammation without any side effects. There is a need for the development of materials that can be used.

In recent years, in order to meet the needs of consumers, research and development of cosmetics using natural raw materials have been actively conducted.

The inventors of the present invention have studied the applicability of cosmetics to various natural products which have not been known so far. As a result, they have studied echinacea (stem, leaf, flower) and prepared extracts therefrom and measured antioxidative and anti- And the results are excellent, it has been found that an efficacy as a cosmetic and external preparation for skin can be expected.

Echinacea purpurea ) is a perennial herb plant of Asteraceae. It is native to North America and grows about 60 to 150 cm in size. Echinacea has a flower and leaf shape similar to Ludvickia. Its leaves are long elliptical, 4 10 cm long, 2 4 cm wide, with serrations on the edges. Flowers of about 10 cm in diameter of magenta come to the ends of stems and branches over June and September. Echinacea is widely grown for ornamental purposes around the world because of its splendid flowers. It is also used as a treatment for colds and various viral infections by enhancing immunity.

Patent Document 1 of Patent Application No. 10-1078888 B1 discloses that Echinacea adiomyces extracts tissue-cultured have cytoprotective effect and cytoprotective effect against stimulation. The cosmetic formulation containing the extract does not induce skin irritation, The present invention relates to a cosmetic composition for protecting the skin against skin damage and improving the skin.

Patent Document No. 10-0431076 B1 discloses a method of mixing antibacterial effect, moisturizing effect, antioxidative activity, antioxidative effect, antioxidative activity, antioxidative activity, antioxidative activity, anti- An anti-inflammatory effect, a free radical scavenging effect, and the like, and has an effect of improving the healing effect of atopic skin.

However, none of the above references disclose that Echinacea purpurea) it had no aboveground (except rootstock, leaves, disclosed for the antioxidant and anti-inflammatory cosmetic composition of flowers). The inventors of the present invention have studied the applicability of cosmetics to various materials that have not been known so far. As a result, it is possible to produce extracts including stem, leaf and flower except for Echinacea root, And the anti-inflammatory effect was excellent.

Echinacea extract is not only cytotoxic but also inhibits the inflammatory precursor cytokines TNF-, IL-6, and IL-1 in a mononuclear cell model that is the subject of inflammatory responses stimulating macrophages with Lipopolyssaccharide (LPS) And inhibits the formation of iNOS, COX-2 and the production of NO, and exhibits a high antioxidative effect. Therefore, the composition containing the same can be applied to cosmetic compositions having antioxidative and anti-inflammatory effects.

KR 10-1078888 B1 (26 Oct 2011) KR 10-0431076 B1 (Apr. 29, 2004)

Accordingly, it is an object of the present invention to provide cosmetic use of a cosmetic composition containing Echinacea stem, leaf, and flower extract, which has not been studied or attempted in the past, and more preferably, Echinacea stem, leaf and flower extract.

The present invention provides a cosmetic composition comprising as an active ingredient at least one extract selected from flowers, leaves, and stems of Echinacea to solve the above problems of the prior art and achieve the above objects.

In the present invention, there is also provided a cosmetic composition characterized by having one or more uses selected from antioxidation, anti-inflammation, and skin barrier strengthening.

Also, the cosmetic composition according to the present invention is characterized in that the Echinacea extract is a 50-95% ethanol solvent extract.

Also, the cosmetic composition of the present invention is characterized in that the Echinacea extract is a 70% ethanol solvent extract.

Also provided are formulations of lotion, gel, water-soluble liquid, cream, essence, water-in-water type, water-in-oil type, or ointment comprising the cosmetic composition of the present invention.

Eekenia (stem, leaf, flower) extract according to the present invention has excellent antioxidative effect, anti-inflammatory effect and skin barrier strengthening effect and is thus very useful as a cosmetic and dermatological external agent.

1 is a graph showing antioxidative activity against Echinacea (stem, leaf, flower) extract and Echinacea (root) extract.
Figure 2 is a graph showing cytotoxicity evaluation in RAW 264.7 cells for Eekenia (stem, leaf, flower) extract.
FIG. 3 is a graph showing the inhibitory effect of NO production on RAW 264.7 cells against Echinacea (stem, leaf, flower) extracts.
4 is a graph showing inhibitory effect of NO production on RAW 264.7 cells against 70% ethanol extract of Echinacea (stem, leaf, flower) and 70% ethanol extract of Echinacea (root).
5 is a graph showing inhibitory effect of PGE2 production on RAW 264.7 cells against 70% ethanol extract of Echinacea (stem, leaf, flower).
FIG. 6 is a graph showing the inhibitory effect of IL-1β production on RAW 264.7 cells against 70% ethanol extract of Echinacea (stem, leaf, flower). FIG.
7 is a graph showing the inhibitory effect of IL-6 production on RAW 264.7 cells against 70% ethanol extract of Echinacea (stem, leaf, flower).
FIG. 8 is a graph showing the inhibitory effect of TNF-α production on RAW 264.7 cells against 70% ethanol extract of Echinacea (stem, leaf, flower).

The present invention relates to Eekenacea (stem, leaf, flower) extract having antioxidant activity and anti-inflammatory activity.

Hereinafter, specific examples of examples and test examples will be described in order to facilitate understanding of the present invention. However, the following examples should not be construed as limiting the invention, and ordinary variations of the person skilled in the art are possible within the scope of the invention.

Example

Example  One - ( Eckenia  (Stem, leaf, flower) of the extract Heat number  The extract and On solvent star Ethanol extract, Eckenia  (Roots) ethanol extract)

Echinacea (stem, leaf, flower) hot water extract was prepared by pulverizing Echinacea (stem, leaf, flower) washed with distilled water in a blender, adding 4 L of distilled water to 40 g of the fine powder sample, heating at 100 ° C. for 3 hours And hot water extraction was performed. After the extraction was performed, the filtrate was filtered through a 400 mesh filter cloth, and the filtrate was lyophilized in a freeze dryer. Echinacea (stem, leaf, flower) ethanol extracts were prepared by dissolving 40 g of microcrystalline powder of Echinacea (stem, leaf, flower) washed with distilled water in 30, 50, 70, And then extracted twice for 3 days. Then, the supernatant was collected, concentrated under reduced pressure, and lyophilized. Echinacea (roots) also produced 70% ethanol extract under the above conditions.

7.1 g of Echinacea (stem, leaf, flower) hot water extract, 3.2 g of 30% ethanol extract of Echinacea (stem, leaf and flower), 5 g of 50% ethanol extract, 4.7 g of 70% % Ethanol extract. In addition, 4.2 g of Echinacea (root) 70% ethanol extract was obtained.

The thus obtained product was used in the following test examples.

Experimental Example  One - Experiment to measure antioxidant effect

Free radical scavenging test was carried out in order to confirm the antioxidative effect of the Eckenia (stem, leaf, flower) extract and Eckenia (root) extract of the present invention by a skin aging improving effect test.

The free radical scavenging test uses the fact that the absorbance of stable DPPH exhibits the maximum absorbance at 540 nm. As the free radical DPPH is erased by the sample and becomes transparent color in purple, that is, as the free radical scavenging rate is increased, And the absorbance of the sample was decreased.

First, 1 mL of 0.1 mM 2,2-diphenyl-1-picryl-hydrazyl radical (DPPH, Sigma) solution was diluted with methanol to a suitable concentration, and the mixture was allowed to stand at 37 ° C for 15 minutes The absorbance at 540 nm was measured using a microplate reader (UVT-06685, Thermo max, USA).

In the free radical scavenging test, 1 mL of DPPH and 1 mL of methanol were added to the control, and 1 mL of methanol and 1 mL of the sample were added to obtain the respective color correction values for the sample and the control.

The free radical scavenging ratio was calculated using the following formula 1, and it was found that the extract of Eckenacea (stem, leaf and flower), the 30% ethanol extract, the 50% ethanol extract, the 70% ethanol The DPPH scavenging activity of the extract and 95% ethanol extracts was 90.7%, 92%, 80.2%, 60.8%, 96.8% and 85.2% at 500 μg / mL, respectively. Especially, 70% ethanol extract, hot water extract and Echinacea (root) 70% ethanol extract of Echinacea (stem, leaf, flower) extracts showed DPPH reduction power as high as ascorbic acid as the control group.

Experimental Example  2 - Cytotoxicity experiment

In order to measure the cytotoxicity of the Eekenacea (stem, leaf, flower) extract prepared in Example 1, RAW 264.7 cells, a macrophage cell line of rats, were purchased from Korean Cell Line Bank.

Antibiotics, Fetal bovine serum (FBS) and Dulbecco's modified eagle's medium (DMEM) were purchased from Gibco / RBL (USA).

The RAW 264.7 cells were cultured in DMEM medium containing 1% antibiotic and 10% FBS at 37 ° C in a 5% CO2 incubator and subcultured once every two days.

RAW 264.7 cells were plated on a 24-well plate at 1.8 × 10 5 cells / mL per well and incubated for 24 h at 37 ° C in a 5% CO 2 incubator. After culturing for 24 hours, the cytotoxicity of Ekenesia (stem, leaf, flower) extracts and hot water extracts was determined by MTT analysis.

The MTT analysis was performed as follows. MTT assay is a representative method for measuring cell survival rate. In living cells in which metabolism is strong, 3- (4,5-dimethylthiaxo-2-yl) -2,5 -diphenyl tetrazolium bromide (MTT) to form a purple, water-insoluble formazan. RAW 264.7 cells were cultured in a 96-well plate at 1.8 × 10 5 cells / mL using DMEM medium supplemented with 10% FBS, cultured for 18 hours, treated with LPS (1 μg / mL) and cultured for 24 hours. Then, 50 μL of MTT solution was added and reacted for 4 hours. After the culture medium was completely removed, 200 μL of dimethylsulfoxide (DMSO) was added to completely dissolve the precipitate, and the absorbance at 540 nm was measured using a microplate reader. The average absorbance value of each sample group was determined and compared with the absorbance value of the control group, the cell growth rate was evaluated.

As shown in FIG. 2, the extract of the extract of Echinacea (stem, leaf, flower) and the hot water extract prepared in Practical Example 1 hardly cause cytotoxicity at a treatment concentration of 200 μg / mL .

Experimental Example  3 - By extraction solvent Nitrogen monoxide  (NO) production inhibitory activity measurement experiment

In order to confirm the anti-inflammatory effect of the extraction solvent of the extract of Echinacea (stem, leaf, flower) prepared in Example 1, the ability to inhibit the production of nitrogen monoxide (NO), which is one of inflammatory substances, was analyzed . RAW 264.7 cells cultured in Experimental Example 2 were used, and samples of extractive solvents of extracts of Echinacea (stem, leaf, flower) prepared in Example were treated with Lipopolysaccharide (LPS, 1 μg / And then the NO production inhibition was analyzed by the Greiss reaction.

The grease reaction is a method of quantitatively analyzing the amount of released NO produced in the culture medium. After 24 hours of sample treatment, 100 μL of the cell culture medium is dispensed into a 96-well plate, and then the grease reaction solution A (0.1% naphthylenediamine dihydrochloride) buffer solution (1: 1) was dispensed into a 96-well plate in an amount of 100 μL each, The absorbance at 540 nm was measured using a plate reader, and the value was expressed as an average value of three repeated experiments.

As shown in FIG. 3, the LPS alone treatment induced NO production, while the iNOS inhibitor, 2-amino-4-methylpyridine, inhibited NO production by 95.7%. The extracts of Echinacea (stem, leaf, flower) extracts showed 70% ethanol extract inhibition of NO production by 51.9% compared to LPS alone treatment. .

Experimental Example  4 - 70%  For ethanol extracts Nitrogen monoxide (NO)  Production inhibition activity measurement experiment

The results of Experimental Example 3 show that 70% ethanol extract of Echinacea (stem, leaf, flower) and Echinacea (root) 70% ethanol extract, Production inhibitory ability.

70% ethanol extract of Echinacea (stem, leaf, flower), 70% ethanol extract of Echinacea (roots) and Lipopolysaccharide (LPS, 1 μg / mL) were treated together to induce NO production, and then the NO production inhibitory activity was analyzed through a Greiss reaction.

The grease reaction is a method of quantitatively analyzing the amount of released NO produced in the culture medium. After 24 hours of sample treatment, 100 μL of the cell culture medium is dispensed into a 96-well plate, and then the grease reaction solution A (0.1% naphthylenediamine dihydrochloride) buffer solution (1: 1) was dispensed into a 96-well plate in an amount of 100 μL each, The absorbance at 540 nm was measured using a plate reader, and the value was expressed as an average value of three repeated experiments.

As shown in FIG. 4, the LPS alone treatment induced NO production and 2-amino-4-methylpyridine, an iNOS inhibitor, inhibited NO production by 95.7%. The 70% ethanol extract of Echinacea (stem, leaf, flower) and the 70% ethanol extract of Echinacea (root) inhibited NO production by 61.1% and 45.3% at the treatment concentration of 300 μg / Flower) 70% ethanol extracts were more effective in inhibiting NO production.

Therefore, the following experiment was conducted to investigate the effect of 70% ethanol extract of Echinacea (stem, leaf, flower) on the expression of inflammatory response factors, which is more effective in inhibiting NO production than Echinacea (root) 70% ethanol extract.

Experimental Example 5 - Prostaglandin  ( PGE2 ) Effect on production

Cyclooxygenase (COX) is an enzyme that converts arachidonic acid into prostaglandins (PGs), classified as COX-1 and COX-2. COX-1 acts on normal vital functions such as platelet formation, gastric wall protection and maintenance of renal function in the body, and COX-2 forms PGE2, an inflammation mediator. PGE ₂ is known to be deeply involved in the development of cancer, such as promoting inflammation, immune response, and angiogenesis.

The ability to inhibit the production of PGE2 was analyzed for the 70% ethanol extract of Echinacea (stem, leaf, flower), which had the highest inhibitory effect on NO production through Experimental Example 4 above. The inhibitory effect of inflammatory PGE2 on rat macrophage RAW 264.7 cells was quantitated using an ELISA kit.

RAW 264.7 cells were inoculated into 24-well plates at 1.8 × 10 5 cells / mL using DMEM supplemented with 10% FBS and cultured for 18 hours. After the medium was removed, 50 μL of the sample prepared at 10 × concentration (1 mg / mL) and 450 μL of the medium containing LPS (1 μg / mL) were simultaneously treated and cultured for 24 hours. After 24 hours, the culture medium was centrifuged (12,000 rpm, 3 min) and the PGE2 content of the supernatant was measured. All samples were stored frozen (-20 ° C) until quantitation. PGE2 was quantitated using a mouse enzyme-linked immunosorbent assay (ELISA) kit (R & D Systems Inc., Minneapolis, MN, USA). The r2 value of the standard curve for the standard was 0.99 or more.

As shown in FIG. 5, the LPS alone treatment group induced the production of PGE2, and the 70% ethanol extract of Echinacea (stem, leaf, flower) produced PGE2 at 100, 200 and 300 μg / 88.1%, 70.8%, and 57.2%, respectively, indicating a concentration-dependent inhibitory effect.

Experimental Example 6-proinflammatory cytokines  ( TNF -α, IL-6, IL- 1β ) Production inhibition activity measurement experiment

In Experiment 4, 70% ethanol extract of Echinacea (stem, leaf, flower) which had the best inhibitory effect on NO production was tested for TNF-α, IL-6 and IL-1β The inhibitory effect was measured using an ELISA kit.

 RAW 264.7 cells were inoculated into 24-well plates at 1.8 × 10 5 cells / mL using DMEM supplemented with 10% FBS and cultured for 18 hours. After the medium was removed, 50 μL of the sample prepared at 10 × concentration (1 mg / mL) and 450 μL of the medium containing LPS (1 μg / mL) were simultaneously treated and cultured for 24 hours. The culture medium was then centrifuged (12,000 rpm, 3 min) to determine the proinflammatory cytokine production of the supernatant. All samples were stored frozen (-20 ° C) until quantitation. The proinflammatory cytokines were quantified using a mouse enzyme-linked immunosorbent assay (ELISA) kit (R & D Systems Inc., Minneapolis, MN, USA).

As shown in FIG. 6, the LPS alone treatment induced the production of IL-1β, and the 70% ethanol extract of Echinacea (stem, leaf, flower) stimulated the production of IL-1β at a concentration of 300 μg / Inhibitory effect and concentration-dependent inhibitory effect.

As shown in FIG. 7, the LPS alone treatment induced the production of IL-6, and the 70% ethanol extract of Echinacea (stem, leaf, flower) inhibited the production of IL-6 by 48.2% at 300 μg / Inhibitory effect and concentration-dependent inhibitory effect.

As shown in FIG. 8, the LPS alone treatment induced the production of TNF-α, and the 70% ethanol extract of Echinacea (stem, leaf, flower) produced TNF-α at a concentration of 300 μg / Inhibitory effect and concentration-dependent inhibitory effect.

Echinacea purpurea is prepared, shredded and pulverized into fine powder, which is immersed in hot water extract and 30%, 50%, 70% ethanol, After the ethanol precipitate was extracted, the supernatant of the extract was recovered, concentrated under reduced pressure and lyophilized.

Accordingly, the inventors of the present invention have confirmed the physiological activity against extracts of Echinacea (stem, leaf, flower) and found the excellent antioxidative and antiinflammatory effects of Echinacea (stem, leaf, flower) . That is, the present invention extracts Echinacea purpurea (stem, leaf, flower) with hot water extract and 30%, 50%, and 70% ethanol to evaluate antioxidative ability through DPPH radical scavenging ability, By observing inflammatory cytokine inhibitory effects, the 70% ethanol extracts of Echinacea (stem, leaf, flower) extracts have shown excellent antioxidant and anti-inflammatory effects.

Inflammation reaction is a normal defense reaction that occurs when a foreign body enters a living body, has metabolic disorder, burns, mechanical or chemical trauma, and detoxifies the pathogen or restores damaged tissue.

The inflammatory response is mediated by macrophages, lymphocytes, polymorphornuclear leukocytes (PMNs), monocytes, mast cells, platelets, fibroblasts, etc. The inflammatory mediators include substances that are released through stimulation by inflammatory reactions, The former is a vasodilating amine released from mast cells. Histamine, cepofine, and the latter are metabolites of arachidonic acid produced from mast cells and PMN, cytokines from monocytes, macrophages and neutrophils And active oxygen species produced.

The macrophage for bacteria produces active oxygen species, PAF, interferon, interleukin, tumor necrosis factor and the like. The active oxygen species among macrophage metabolites exhibit bactericidal action in the cell gap, while arachidonic acid (arachidonic acid) metabolism to increase the production of chemical agents and PGs, thereby accelerating the production of reactive oxygen species at the site of inflammation.

At this time, superoxide dismutase (SOD), which is a superoxide anion degrading enzyme in the intercellular space, and catalase (CAT), a degradative enzyme of H ₂ O ₂ , At low levels, the susceptibility of the intercellular components to the superoxide anion is increased, promoting the production of superoxide anion-dependent inflammatory mediators and hydroxyl radicals at the site of inflammation, resulting in the formation of neutrophils and neutrophils around the inflammatory mediators The same granulocyte concentration becomes excessive.

This condition can cause damage to normal cells, such as the destruction of biomacromolecules in the cell gap, peroxidation of tissue membrane lipids, edema due to increased cell membrane fluidity, and the like, by propagating the inflammatory response.

Therefore, in the present invention, a free radical scavenging test was conducted in order to confirm the antioxidative effect of the extract of Eckenia (stem, leaf, flower) by the skin aging improving effect test.

As a result, the free radical scavenging activity of DPPH used in the measurement of electron donating ability was higher than that of 70% ethanol extract of Echinacea (stem, leaf, flower) and 30% and 50% ethanol extract ) Scavenging activity (Fig. 1).

Although normal NO formation plays an important role in killing bacteria or eliminating tumors, NO produced by iNOS in the inflammatory state not only promotes inflammatory responses such as vascular permeability and edema, but also stimulates inflammation mediator biosynthesis, It is known to intensify.

We examined the effects of Echinacea (stem, leaf, flower) extracts on the production of nitric oxide (NO), one of the active oxygen species and recently known to play an important role in inflammation induction.

The results showed that 70% ethanol extract of Echinacea (stem, leaf, flower) showed higher NO production inhibitory effect than LPS alone treatment, and that the hot water extract and 30% and 50% (Fig. 3, 4).

PGE ₂ is known to be deeply involved in the development of cancer, such as promoting inflammation, immune response, and angiogenesis. In order to investigate whether 70% ethanol extract of Echinacea (stem, leaf, flower), which had excellent inhibitory effect on NO production, was involved in the expression of inflammatory PGE ₂ in RAW 264.7 cells, it was quantified using ELISA kit. RAW 264.7 cells were treated with LPS (1 μg / mL) to induce the production of PGE ₂ , followed by treatment with 70% ethanol extract of Echinacea (stem, leaf, flower) at a concentration of 300 μg / mL or less. Dependent inhibition of PGE2 production (Fig. 5).

Lipopolysaccaride (LPS), also known as endotoxin, is present in the extracellular membrane of Gram-negative bacteria and has been shown to inhibit tumor necrosis factor (TNF- [alpha]) in macrophages or monocytes such as RAW 264.7 cells, IL-6 (Interleukin-6), and IL-1β (Interleukin-1β) are known to increase proinflammatory cytokines.

Thus, the expression level of pro-inflammatory cytokines from RAW 264.7 cells was quantitated by ELISA kit.

 As a result, 70% ethanol extract of Echinacea (stem, leaf, flower) for inhibition of IL-1β, IL-6 and TNF-α production by treatment with LPS stimulation, 1β to 42.8%, IL-6 to 48.2% and TNF-α to 65.1% (FIGS. 6 and 7. 8).

From the above experimental results, it was confirmed that the extract of Echinacea (stem, leaf, flower) of the present invention has excellent antioxidative activity and anti-inflammatory activity.

Example  2, and Comparative Example  One - Eckenia  Cream formulation containing the extract (formulation)

The components of the cream containing the 70% ethanol extract of Echinacea (stem, leaf, flower) or the 70% ethanol extract of roots obtained in Example 1 were prepared as shown in Table 1 below. The units shown in Table 1 are wt%.

number ingredient Example 2 Example 3 Comparative Example 1 One Pro-type glycerin monostearate 2.0 2.0 2.0 2 Stearic acid 1.5 1.5 1.5 3 Cetearyl alcohol 2.2 2.2 2.2 4 Wax 1.0 1.0 1.0 5 Squalane 3.0 3.0 3.0 6 Hardened vegetable oil 1.0 1.0 1.0 7 Sorbitan stearate 0.6 0.6 0.6 8 Mineral oil 5.0 5.0 5.0 9 Polysorbate 60 1.5 1.5 1.5 10 Dimethicone 1.0 1.0 1.0 11 Trioctanoin 5.0 5.0 5.0 12 Betaine 3.0 3.0 3.0 13 Triethanolamine 1.0 1.0 1.0 14 glycerin 5.0 5.0 5.0 15 Sodium hyaruronate 3.0 3.0 3.0 16 Echinacea (stem, leaf, flower) 70% ethanol extract 1.0 - - 17 Echinacea (root) 70% ethanol extract - 1.0 - 18 Distilled water Balance Balance Balance 19 Preservative, incense a very small amount a very small amount a very small amount

The aqueous components of the raw materials 12 to 15 and 17 were completely dissolved by heating to 80 ° C among the components constituted by the composition of Table 1 and then the raw materials 1 to 11 and 16 were heated to 80 ° C to obtain the raw materials 12 to 15 and 17 (Homo Mixer Mark II, Primix, JPN) for 15 minutes at 3,000 rpm. Thereafter, the raw material 18 was added, stirred for 5 minutes, and then cooled to room temperature.

Experimental Example  7 - Assessment of skin primary stimulation

The evaluation test of the human skin primary stimulus of the cream prepared in Example 2 was performed by P & K, a professional clinical testing institute. The subjects were selected from more than 30 adults who did not meet the selection criteria and who were not eligible under the exclusion condition. The subjects were flattened except for the vertebrae and applied to areas where there was no pigmentation or skin damage. Skin irritation was assessed by the Frosch & Kligman, CTFA (Cosmetic, Toiletry and Fragrance Association) guideline and the skin reaction was evaluated using the following equation (2) according to the criteria given in Table 2 below. The skin irritation information of the test water was also determined by referring to the skin irritation index table of Table 3 using the following Equation 3.

sign Rating Criteria + One Some erythema, spots or diffuse ++ 2 Uniform erythema of the degree of increase +++ 3 Severe erythema with edema ++++ 4 Severe erythema with edema and small bubbles

Skin irritation index division Remarks 0.00 to 0.25 Non-irritant 6.25% 0.26 to 1.00 Weak irritation 25.00% 1.01 to 2.50 Moderate irritancy 62.50% 2.51 to 4.00 Strong irritant 100.00%

As shown in the following Table 4, the cream of Example 2 was applied for 24 hours, and the skin reaction at 1 hour and 24 hours after the removal of the patch was visually evaluated by a researcher. The skin irritation index and the degree of skin irritation were observed, Respectively.

Test Products Skin irritation index Degree of skin irritation Example 2 0.02 Non-irritant Example 3 0.11 Non-irritant

Experimental Example  8 - Skin barrier  Strengthening effect measurement

Measurement of transepidermal water loss (TEWL), which measures water loss through the stratum corneum from the dermis, is commonly used to evaluate the stratum corneum status and to evaluate skin barrier function. The moisture content was measured by measuring the capacitance according to the moisture content, and the relative size of the moisture content was measured.

The skin barrier enhancement measurement of the cream prepared in Example 2 was performed by P & K, a specialized clinical testing institution. The subjects were selected more than 30 adults who satisfied the selection criteria and did not have the exclusion condition. The measurement of the TEWL before the use of the Example 2 and the Comparative Example 1 and the measurement of the TEWL after the induction of the SLS injury Vapor mass loss (TEWL) of the vapometer was measured.

As shown in the following Table 5, the skin TEWL measurement value was 16.32 ± 11.27 at the first measurement and 15.48 ± 9.62 after 2 weeks in Comparative Example 1 in which the 70% ethanol extract of Echinacea (stem, leaf, flower) was not contained, The formulation containing 70% ethanol extract of nessia (stem, leaf, flower) in Example 2 was 17.22 ± 5.60 at the first measurement and 10.85 ± 10.11 after 2 weeks. The average change in TEWL before and after use of the cream was observed to be -0.84 ± 8.53 in the control product and -6.37 ± 5.67 in the test product and decreased after 2 weeks of using the test product. This reduced water loss after 2 weeks of use, indicating that skin barrier enhancement was achieved by reducing moisture loss by using the cream of Formulation Example 2 containing 70% ethanolic extract of Echinacea (stem, leaf, flower) to be.

Experiment 0 weeks 2 weeks Improvement effect TEWL Comparative Example 1 16.22 + - 11.2 15.48 ± 9.62 -0.84 + -8.53 Example 2 17.22 + - 5.60 10.85 ± 10.11 -6.37 ± 5.67 Example 3 16.45 + - 10.60 13.21 + - 8.62 -3.24 + - 6.72

Examples of formulations of the present invention are a soft lotion, a convergent lotion, a nutritional lotion, a massage cream, an essence and a pack, but the formulation of the cosmetic composition of the present invention should not be construed as being limited thereto. Lt; / RTI &gt; is possible.

Formulation Example  1 - softening longevity

number ingredient Content (% by weight) One glycerin 5.00 2 1,3-butylene glycol 3.00 3 PEG 1500 1.00 4 Allantoin 0.10 5 DL-Panthenol 0.30 6 EDTA-2Na 0.02 7 Benzophenone-9 0.04 8 ethanol 10.00 9 Octidodeces-16 0.20 10 Polysorbate 20 0.20 11 Ekenesia (stem, leaf, flower)
70% ethanol extract 5.0 12 Preservative, fragrance, pigment a very small amount 13 Distilled water Balance

Formulation Example  2 - convergent lotion

number ingredient Content (% by weight) One glycerin 2.00 2 1,3-butylene glycol 2.00 3 Allantoin 0.10 4 DL-Panthenol 0.30 5 EDTA-2Na 0.02 6 Benzophenone-9 0.04 7 ethanol 15.00 8 Polysorbate 20 0.20 9 Ekenesia (stem, leaf, flower)
70% ethanol extract 1.0 10 Citric acid a very small amount 11 Preservative, fragrance, pigment a very small amount 12 Distilled water Balance

Formulation Example  3 - Nourishing lotion

number ingredient Content (% by weight) One Cetearyl alcohol 1.00 2 Glyceryl stearate 0.50 3 Polysorbate 60 1.00 4 Sorbitan sesquioleate 0.30 5 Cetyl octanoate 6.00 6 Squalane 4.00 7 Shapower Oil 4.00 8 Butylene glycol 4.00 9 glycerin 4.00 10 Carbomer 0.10 11 Triethanolamine 0.10 12 Ekenesia (stem, leaf, flower)
70% ethanol extract 1.00 13 Preservative, fragrance, pigment a very small amount 14 Distilled water Balance

Formulation Example  4 - essence

number ingredient Content (% by weight) One glycerin 10.00 2 Betaine 5.00 3 PEG 1500 2.00 4 Allantoin 0.10 5 DL-Panthenol 0.30 6 EDTA-2Na 0.02 7 Benzophenone-9 0.04 8 Hydroxyethylcellulose 0.10 9 Carboxyvinyl polymer 0.20 10 Triethanolamine 0.18 11 Octyldodecanol 0.30 12 Octyldodec-16 0.40 13 ethanol 6.00 14 Ekenesia (stem, leaf, flower)
70% ethanol extract 1.00 15 Preservative, fragrance, pigment a very small amount 16 Distilled water Balance

Formulation Example  5-pack

number ingredient Content (% by weight) One Polyvinyl alcohol 15.00 2 Cellulose sword 0.15 3 glycerin 3.00 4 PEG 1500 2.00 5 Sheik Destrin 0.15 6 DL-Panthenol 0.30 7 Allantoin 0.10 8 Monoammonium glycyrrhizin acid 0.20 9 Nicotinamide 0.40 10 ethanol 5.00 11 PEG 40 hardened castor oil 0.30 12 Ekenesia (stem, leaf, flower)
70% ethanol extract 1.00 13 Preservative, fragrance, pigment a very small amount 14 Distilled water Balance

※ This invention is the result of the research of the leading industrial development business of the metropolitan economy carried out with the support of the Ministry of Commerce, Industry and Energy, the Korea Industrial Technology Development Agency, and the Jeju Regional Business Evaluation Institute. (This invention was financially supported by Ministry of Trade, Industry and Energy (MOTIE), the Korea Institute for Advancement of Technology (KIAT), and the Jeju Institute for Regional Program Evaluation through the Leading Industry Development for Economic Region.

Claims (5)

A cosmetic composition comprising as an active ingredient at least one topically-derived extract selected from flowers, leaves, and stems of Echinacea to prevent or improve dryness due to skin weakness through skin barrier enhancement. delete [Claim 4] The cosmetic composition according to claim 1, wherein the Echinacea extract is a 50-95% ethanol solvent extract. [Claim 4] The cosmetic composition according to claim 3, wherein the Eicinia extract is a 70% ethanol solvent extract. A formulation of a cosmetic, a gel, a water-soluble liquid, a cream, an essence, an underwater type formulation, a water-in-oil type formulation, or an ointment comprising the cosmetic composition of claim 1, claim 3 or claim 4.

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