KR20160008942A - A composition comprising fermented Myrothamnus flabellifolius extract and the use thereof - Google Patents

Abstract

The present invention relates to a cosmetic composition for skin whitening, skin wrinkle improvement, skin elasticity improvement or antioxidation comprising a fermented extract of Echinochloa as an active ingredient, a pharmaceutical composition for antioxidation, As an active ingredient, to a pharmaceutical composition for preventing or treating an inflammatory disease.

Description

[0001] The present invention relates to a composition comprising fermented Echinochloa crus-galli extract and to its use.

The present invention relates to a cosmetic composition for skin whitening, skin wrinkle improvement, skin elasticity improvement or antioxidation comprising a fermented extract of Echinochloa as an active ingredient, a pharmaceutical composition for antioxidation, As an active ingredient, to a pharmaceutical composition for preventing or treating an inflammatory disease.

The skin is an important body that is responsible for various physiological functions such as barrier function, body temperature control function, excretion function, which protects the human body from the external environment and prevents the internal moisture and useful components from flowing out. However, due to the following reasons, the activity of the skin cells may be lowered, and the skin may develop changes such as spots, freckles, and black blotches, and the skin may become dull and rough.

First, the collagen existing in the dermal layer of the skin performs many functions such as mechanical rigidity of the skin, resistance of the connective tissue, maintenance of the bonding force of the tissue, and support of the cell adhesion, and thus has a great influence on the constituents of the skin. Such collagen is reduced by photo aging due to stress, aging, or ultraviolet irradiation, thereby loosening the skin tissue and losing its elasticity.

Second, inflammation reaction occurs by external factors such as stress or ultraviolet ray irradiation, or activation of immune cells is reduced by aging, so that control of immune function of the skin becomes non-qualitative, thereby promoting skin aging.

Third, the cell function of the epidermis is deteriorated, the metabolism is not smooth, and the exfoliation does not occur well, and when the ultraviolet light is received in the state where the keratin is accumulated in an excessive amount, the elasticity is lowered and wrinkles are generated.

Fourth, if no epidermis is formed on the epidermis, moisture and sebum secretion decrease, and the skin becomes dry and wrinkles are formed.

Fifth, oxygen radicals generated from subcutaneous fat break down cell membranes and nuclei to lower blood circulation, and skin nutrition and moisture become insufficient and skin aging is promoted.

Sixth, melanin is synthesized to protect skin when skin receives ultraviolet rays. The synthesized melanin is transferred to the keratinocytes of the skin through the melanoma. This keratinocyte turns over over a period of 28 days. Therefore, the melanin produced is generally lost by the keratinocyte every 28 days, but when the skin cell cycle is not controlled by the stress and skin aging, the keratinocyte does not fall out in the period, Pigmentation of the skin occurs and skin aging occurs.

Seventh, in skin, sebum, sweat and cosmetic ingredients can be decomposed into substances that are toxic to skin by the fungus, causing skin irritation and inflammation. In addition, stimulation by ultraviolet rays can cause inflammation by increasing the production of nitrogen monoxide (NO), an inflammation mediator, in the skin. The production of nitrogen monoxide is mostly caused by iNOS, and it is known that iNOS is rapidly induced by stimulation such as LPS and cytokine to produce excessive NO.

Various studies have been conducted on a moisturizing agent, a whitening agent, and a cosmetic for delaying skin aging to improve skin wrinkles and inhibit skin pigmentation. Furthermore, as an aging society, consumers' desire to have a young and elastic skin has increased. In July 2000, the Cosmetic Act was enacted to research and develop cosmetics that delay skin aging and improve skin moisturization. More active. Cosmetics of this nature also use natural materials to avoid toxicity or irritation to the skin.

The Resurrection is also called the rose of Jericho because it grows in the Jericho valley in the Bible, and the included trihalos helps the resurrection to live in extreme environments. Trihalose contains hydroxy (OH) groups in its molecule and can attract a lot of water, so it is known that it has an excellent effect in enhancing skin moisturizing power.

Under these circumstances, the inventors of the present invention have made extensive efforts to develop a method for inducing skin whitening, wrinkle improvement, and skin elasticity improvement. As a result, fermented Echinochloa extract has not only a skin whitening effect, Anti-inflammatory and antioxidative effects, and completed the present invention.

The main object of the present invention is to provide a cosmetic composition for skin whitening, prevention or improvement of skin wrinkles, improvement of skin elasticity, or antioxidation, which contains fermented extract of Echinochloa as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating an inflammatory disease, which comprises at least one selected from the group consisting of fermented extracts of Echinochloa crus-galli and extracts of Echinochloa as an active ingredient.

It is still another object of the present invention to provide a pharmaceutical composition for antioxidation, which contains fermented extract of Echinochloa as an active ingredient.

Cosmetics  Composition

As one embodiment for achieving the above object, the present invention provides a cosmetic composition for skin whitening, preventing or improving skin wrinkles, improving skin elasticity, or antioxidation comprising fermented extract of Echinochloa as an active ingredient.

The skin whitening refers to inhibiting melanin formation and inhibiting melanin formation. As a result of measuring the amount of melanin produced by treating melanoma cells of rats with fermented resurrected extracts, the extract of fermented resurrected herb was found to be superior to arbutin, which is known to inhibit melanin production by acting directly on tyrosinase (Example 4, Table 4). In addition, the present invention provides a method for inhibiting melanin formation, comprising the steps of:

The skin wrinkles are caused by the skin of the skin and can be caused by the cause of the gene, collagen and elastin present in the skin dermis, and the external environment. The prevention or improvement of skin wrinkles according to the present invention refers to inhibiting or inhibiting the generation of wrinkles on the skin or alleviating the wrinkles already formed.

The skin elasticity refers to elastic fibers composed of elastin present in the dermal layer and collagen fibers called collagen. The skin elasticity improvement means that the skin elasticity is maintained in a state in which elastin and collagen are sufficiently present .

The inventors of the present invention have found that collagen synthesis is superior to vitamin C, which is involved in collagen synthesis, and that collagen synthesis is promoted more effectively than collagen synthesis Examples 5 and 6, Tables 5 and 6).

The antioxidant refers to inhibition of cellular oxidation by free radicals or reactive oxygen species (ROS), which are highly reactive according to oxidative stress caused by intracellular metabolism or ultraviolet rays, and free radicals Or removal of reactive oxygen species, thereby reducing damage to the cells. The inventors of the present invention found that the fermented extract of Echinochloa extract had a strong free radical scavenging ability even when compared with vitamin C and that it was superior to the fermented Echinochloa extract in that it was a natural extract, , Table 7).

The fermentation of the fermented extract of Echinochloa can be carried out by methods well known in the art. For example, the fermentation may be a fermentation by inoculating a fermented microorganism into a dried product, a pulverized product, a concentrate, or a mixture thereof. In another example, the fermentation can be carried out by adding a dried product, a concentrate, or a mixture thereof to a culture medium of a microorganism, then inoculating the fermentation microorganism into the medium and fermenting the mixture at a suitable temperature and humidity for a suitable period of time have. For example, the fermenting microorganism may be at least one selected from the group consisting of mushroom, lactic acid bacteria, and yeast.

The mushroom used for the fermentation may be, for example, a shiitake mushroom, a mushroom mushroom, a mushroom mushroom, a mushroom mushroom, a mushroom mushroom, a mushroom mushroom, a mushroom mushroom, a mushroom or a mushroom, Prevention or improvement, skin elasticity improvement, or antioxidative effect.

The lactic acid bacteria may be, for example, Bifidobacterium sp.), Lactobacillus genus (Lactobacillus sp.), Lactococcus lactis in (Lactococcus sp.), Peddie Oh caucus in (Pediococcus sp.), Streptococcus genus (Streptococcus. sp), or the current Kono Stock in (Leuconostoc sp.). However, the present invention is not limited to the above, which can exhibit skin whitening, skin wrinkle prevention or improvement, skin elasticity improvement, or antioxidative effect. For example, the lactic acid bacteria may be selected from the group consisting of Bifidobacterium bifidum , B. breve , B. longum , Bifidobacterium animaris ( B. animalis), Bifidobacterium lactis (B. lactis), Lactobacillus ash's also a filler (L. acidophilus), Casey Lactobacillus (L. casei), Lactobacillus joined Lee (L. gasseri), Lactobacillus del Brewer L. delbrueckii spp., L. bulgaricus , L. helveticus , L. fermentum , L. paracasei , L. plantarum , L. reuteri , L. rhamnosus , L. salivarius , L. lactis , and the like), Lactobacillus spp ., Lactobacillus sp. , Peddie Oh Celebi jiae Caucus (P. cerevisiae), Peddie Oh Caucus kids CD Rock City (P. acidilactici), Streptococcus lactis (S. lactis), Streptococcus thermostat Phillies (S. thermophiles), Streptococcus Crescent Morris (S. cremoris), or the current mail center Stock Pocono Roy Rhodes (L. mesenteroides) Lt; / RTI > In the present invention, lactic acid bacteria may be used in the same sense as lactic acid bacteria.

The yeast may be, for example, Saccharomyces cerevisiae , S. ellipsoideus , S. coreanus , Saccharomyces cylindusensis (" Saccharomyces cerevisiae " S. carlsbergensis), Saccharomyces access to my Paz Astoria Augustine (S. pastorianus), Saccharomyces access to my lactis (S. lactis), Saccharomyces access to my ruksi (S. rouxii), to access Shijo Saccharomyces pombe Mai (Schizosaccharomyces pombe ), Zygosaccharomyces major major , Hansenula anomala , Brettanomyces bruxellensis , B. custersianus , Dekkera, anomala , Streptomyces olivochromogenes , or S. griseus . However, the present invention is not limited to skin whitening, skin wrinkle prevention or improvement, skin elasticity improvement, or antioxidative effect The present invention is not limited thereto. For example, the yeast may be Saccharomyces cerevisiae.

The amount of microorganism inoculated, the incubation temperature, the incubation humidity, and the incubation time for microorganism fermentation of the present invention may vary depending on various conditions depending on the kind of microorganism used for fermentation. As an example, the fermented extract of Echinochloa sp. May be obtained by inoculating yeast into a medium containing the resurrected sucrose, or by fermentation at pH 5.5 to 6.5 at 10 to 35 ° C for 24 hours to 5 days. As another example, the fermented extract of Echinochloa may be obtained by inoculating lactic acid bacteria into a culture medium containing the resveratrol or a medium containing the same, and fermenting at a pH of from 6.5 to 7.5 and a temperature of from 35 to 38 ° C for 24 hours to 5 days. As another example, the fermented extract of Echinacea can be obtained by inoculating the bifidobacteria into the medium containing the resurrected supernatant or the fermented bifidus and fermenting it for 24 hours to 5 days while maintaining the anaerobic culture conditions at a pH of 6.5 to 7.5 and a temperature of 35 to 38 ° C . ≪ / RTI >

The fermented extract of Echinochloa can be prepared from the Echinochloa fermented by methods well known in the art. For example, the fermented activated syrup may be finely ground according to a conventional method, and water, an anhydrous or a lower alcohol having 1 to 4 carbon atoms (such as methanol, ethanol, propanol and butanol), glycerin, Glycol, and 1,3-butylene glycol; and hydrocarbon solvents such as chloroform, methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether, and dichloromethane, Can be used.

As another example, the fermented extract of Eucalyptus may be water and ethanol extract of fermented Eucalyptus. For example, the ratio of water to ethanol may be 1: 1 to 1:10 by volume, for example, 1: 1 to 1: 5 by volume.

The amount of the extraction solvent may vary depending on the kind of the extraction solvent used, but may be, for example, 1 to 20 times, or 5 to 20 times the dry weight of the fermentation product. The extraction may be performed one or more times, and the extract obtained by one or more extractions may be mixed and concentrated under reduced pressure. In addition, the fermentation product may be obtained by a drying process (for example, drying at 40 to 70 ° C for 15 to 50 hours) before pulverization, followed by pulverization and extraction.

The fermented extract of Echinochloa sp. May contain not only the extract obtained by the above-mentioned extraction solvent, but also an extract obtained through a conventional purification process. In addition, the fermented extract of Echinochloa extract of the present invention can be prepared in powder form by an additional process such as vacuum distillation and freeze-drying or spray-drying.

In another embodiment, the composition may further comprise an extract of Echinochloa sp. As an active ingredient. For example, the resurrected extract may be water and ethanol extract of the resurrection herb.

The fermented extract of Echinochloa or the fermented Echinochloa extract and the Echinochloa extract may be contained in an amount of 0.0001% by weight to 30% by weight or 0.005% by weight to 10% by weight based on the total weight of the whole composition. For example, 0.01% by weight to 5% by weight. As a result of the experiment, when the content is less than 0.0001% by weight, substantial skin antioxidative effect and skin moisturizing effect are difficult to expect. If it exceeds 30% by weight, the manufacturing cost may be too high compared to the cosmetic effect.

The cosmetic composition may be prepared in the form of a general emulsified formulation and a solubilized formulation. For example, creams, essences, serums, cosmetic ointments, sprays, oil gels, gels such as lotions such as lotion, facial lotion, body lotion and the like such as flexible lotion or nutrition lotion, nutrition cream, A lotion, a cleansing lotion, a makeup remover such as a cleansing oil, a cleansing foam, a soap, a body wash and the like such as a foundation, a pack, a sunscreen, a makeup base, a liquid type, a solid type or a spray type, Lt; / RTI >

In addition to the resurfacing extract of the present invention, the fermented resurrection extract or the mixture thereof, the number of softening times may be 1 to 10% by weight of polyhydric alcohol such as propylene glycol or glycerin and 0.05 to 5% by weight of a surfactant such as polyethylene oleyl ether or polyoxyethylene hydrogenated castor oil To 2% by weight.

The nutritional lotion or nutritional cream may contain 5 to 20% by weight of an oil such as squalane, petrolatum, and octyldodecanol in addition to the resurfacial extract, the fermented resurrected extract or the mixture of the present invention, and an antioxidant such as cetanol, stearyl alcohol, And 3 to 15% by weight of wax.

The massage cream may be prepared so as to contain 30 to 70% by weight of an oil such as liquid paraffin, petrolatum, isononylisononanoate and the like in addition to the resurrected extract of the present invention, the fermented resurrection extract or the mixture thereof.

The essence may be prepared so as to contain 3 to 30% by weight of a polyhydric alcohol such as propylene glycol or glycerin, in addition to the extract of the present invention, fermented retort extract or a mixture thereof.

The pack may contain at least one selected from the group consisting of kaolin, talc, zinc oxide, zirconium oxide, and zirconium oxide in a peel off pack or a general emulsion type cosmetic composition containing 5 to 20% by weight of polyvinyl alcohol, And a wash off pack containing 5 to 30% by weight of a pigment such as titanium dioxide.

In addition, the cosmetic composition of the present invention may further contain, in addition to the components described above, the components described above, such as oil, water, a surfactant, a moisturizer, a lower alcohol having 1 to 4 carbon atoms, a thickener, a chelating agent, Perfume and the like can be appropriately mixed and used as needed.

Pharmaceutical composition for preventing or treating inflammatory diseases

As another embodiment for achieving the above object, the present invention provides a pharmaceutical composition for preventing or treating an inflammatory disease, comprising at least one selected from the group consisting of fermented extracts of Echinochloa crus-galli and extracts of Echinochloa as an active ingredient do.

The above-described fermented Easter candle and its extract are as described above.

The ratio of each extract may be in the range of 1: 1 to 10: 1, and in another example 1: 1 to 5: 1, if it includes both the fermented extract and the extract.

The inflammatory disease may be selected from the group consisting of dermatitis, allergic inflammatory diseases including atopy, conjunctivitis, periodontitis, rhinitis, otitis, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, gout, ankylosing spondylitis, rheumatic fever, inflammatory bowel disease, fibromyalgia, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, shoulder inflammation, nephritis, hay fever, tendinitis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis or acute and chronic inflammatory diseases It is not limited.

Nitric oxide (NO) and nitric oxide synthase (NOS) are the major factors involved in the inflammatory response. Oxidative stress caused by NO activates the NF-κB (Nuclear Factor Kappa B) in the cell and accelerates the inflammatory reaction because activated NF-κB enters the nucleus and promotes the expression of an inflammatory gene. NO is mainly produced in macrophages and monocytes, and macrophages are rapidly induced by inflammatory agents such as lipopolysaccharide (LPS). L-arginine is oxidized by NOS to produce NO. Inducible nitric oxide synthase (iNOS) among NOS is closely related to overproduction of NO in cells It is known.

As a result of measuring the inhibitory activity of nitric oxide (NO) in the fermented Echinochloa or Echinochloa extract, it was confirmed that NO production can be effectively inhibited in spite of LPS treatment. In particular, it was confirmed that the NO inhibitory activity of the extract of the resurrected extract fermented was higher than that of the resurrected extract (Experimental Example 8, Table 8). Therefore, the pharmaceutical composition according to the present invention can alleviate the inflammation caused by an increase in NO production, which is an inflammation mediator.

As used herein, the term "prevention" may refer to any action that inhibits or delays the onset of an inflammatory disease by administering the pharmaceutical composition of the present invention to a subject.

The term "treatment" used in the present invention may mean all actions that cause the symptom of an inflammatory disease to be improved or benefited by administering the pharmaceutical composition of the present invention to a suspect subject having an inflammatory disease.

The pharmaceutical composition of the present invention can be used as a single preparation or can be manufactured as a complex preparation containing a drug known to have an approved inflammatory disease effect and can be formulated by using a pharmaceutically acceptable carrier or excipient, ≪ / RTI > or by intrusion into a multi-dose container.

As used herein, the term "pharmaceutically acceptable carrier" may mean a carrier or diluent that does not disturb the biological activity and properties of the compound being injected, without irritating the organism. The type of the carrier that can be used in the present invention is not particularly limited, and any carrier conventionally used in the art and pharmaceutically acceptable may be used. Non-limiting examples of the carrier include saline, sterilized water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol and the like. These may be used alone or in combination of two or more.

In addition, if necessary, other conventional additives such as an antioxidant, a buffer and / or a bacteriostatic agent can be added and used. A diluent, a dispersant, a surfactant, a binder, a lubricant, Pills, capsules, granules or tablets, and the like.

In addition, the pharmaceutical composition of the present invention may contain a pharmaceutically effective amount of fermented extract of Eucalyptus, Eucalyptus extract, or a mixture thereof. The term "pharmaceutically effective amount" as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and includes an extract of the fermented extract, an extract of the extract, , It may mean 0.0001% to 30%, 0.005% to 10%, or 0.01% to 5% by weight based on the total weight of the total composition. As a result of the experiment, if the content is less than 0.0001 wt%, a substantial effect is not expected. If the content is more than 30 wt%, the manufacturing cost may be excessively increased compared to the pharmacological effect. For purposes of the present invention, however, the specific therapeutically effective amount for a particular patient will depend upon the nature and extent of the reaction to be achieved, the particular composition, including whether or not other agents are used, the age, weight, Sex and diet of the patient, the time of administration, the route of administration and the rate of administration of the composition, the duration of the treatment, the drugs used or concurrently used with the specific composition, and similar factors well known in the medical arts.

The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with another therapeutic agent, and may be administered sequentially or simultaneously with conventional therapeutic agents. And can be administered singly or multiply. It is important to take into account all of the above factors and to administer an amount that can achieve the maximum effect in a minimal amount without causing side effects, and can be readily determined by those skilled in the art.

The term "administering" as used herein means introducing the pharmaceutical composition of the present invention to a patient by any suitable method, and the route of administration of the composition of the present invention is not limited to a variety of oral or parenteral routes Lt; / RTI >

The mode of administration of the pharmaceutical composition according to the present invention is not particularly limited and may be conventionally used in the art. As a non-limiting example of such a mode of administration, the compositions may be administered orally or parenterally. The pharmaceutical composition according to the present invention can be manufactured into various formulations according to the intended administration mode.

The dosage of the pharmaceutical composition of the present invention can be administered, for example, to a mammal including a human in an amount of 1 to 20 mg / kg, more preferably 1 to 10 mg / kg per day for the pharmaceutical composition of the present invention , The frequency of administration of the composition of the present invention is not particularly limited, but it may be administered once a day or divided into several doses.

The formulation of the pharmaceutical composition may be, for example, but not limited to, an external preparation for skin such as ointments, patches, gels, creams or sprays. The external preparation for skin may further contain at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickening and gelling agents, softening agents, antioxidants, suspending agents, stabilizing agents, foaming agents, fragrances, surfactants, As well as any other ingredients conventionally used in emulsifying agents, fillers, sequestering and chelating agents, preservatives, vitamins, barrier agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, Adjuvants commonly used in the field of dermatology.

In another aspect, the present invention provides a method of preventing or treating an inflammatory disease, comprising administering the pharmaceutical composition to a subject.

As used herein, the term "individual" may refer to any animal, including a human, who is or may be at risk of developing an inflammatory disease. The animal may be, but is not limited to, a mammal such as a cow, a horse, a sheep, a pig, a goat, a camel, a nutrient, a dog, a cat,

The preventive or therapeutic method of the present invention may specifically include administering the composition in a pharmaceutically effective amount to a subject suffering from or at risk of developing an inflammatory disease.

The term "administering" as used herein refers to the introduction of a pharmaceutical composition of the present invention to a patient by any suitable method, and the route of administration of the composition of the present invention may be oral or parenteral May be administered via various routes.

Antioxidant pharmaceutical composition

In another aspect of the present invention to achieve the above object, the present invention provides a pharmaceutical composition for antioxidation comprising fermented extract of Echinochloa as an active ingredient.

The fermented resurrection extract is as described above.

Antioxidation is the inhibition of cellular oxidation by reactive radicals or reactive oxygen species (ROS), which is highly reactive with oxidative stress caused by intracellular metabolism or ultraviolet radiation. It is the result of removing free radicals or reactive oxygen species Lt; RTI ID = 0.0 > damage. ≪ / RTI >

The inventors of the present invention found that the fermented extract of Echinochloa extract had a strong free radical scavenging ability even when compared with vitamin C and that it was superior to the fermented Echinochloa extract in that it was a natural extract, , Table 7).

Fermented Resurrection  Method of producing extract

According to another aspect, the present invention provides a method for producing fermented milk, which comprises fermenting an agar with at least one selected from the group consisting of mushroom, lactic acid bacteria and yeast; And extracting the fermented resurrection gel with water and ethanol. The present invention also provides a method for producing fermented resurrected extract.

The fermentation method and the extraction method are as described above.

The extract according to the present invention is excellent in anti-inflammation, moisturizing, whitening, wrinkle improvement, elasticity improvement, antioxidant effect, and does not cause allergic reaction or irritation to the skin, It can be used as medicines or cosmetics.

Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited by the following examples.

Comparative Example : Resurrection  Preparation of extract

200 g of the extract was mixed with distilled water and 70% ethanol in a volume ratio of 3: 7, and the mixture was extracted at room temperature (20 [deg.] C) ~ 25 < 0 > C) for 5 hours and then subjected to primary filtration. Then, ethanol was removed from the filtered extract through evaporation, and the precipitate was removed by centrifugation and secondary filtration. Finally, micro-filtration was conducted to obtain 23 g of an extract of Echinochloa sp.

Example : Fermented Resurrection  Preparation of extract

Yeast ( Saccharomyces cerevisiae , purchased from Rady) was inoculated in a YM broth and cultured in a shaking incubator at 30 ° C for 24 hours.

The strains were inoculated in 100% by weight of pulverized resurrection supernatant in a concentration of 59.4% by weight of purified water, 40% by weight of butylene glycol and 0.1% by weight of trehalose, and then cultured in a fermenter at 30 캜 for 48 hours. Thereafter, the fermented product was sterilized with an autoclave and extracted, separated and purified in the same manner as in the above Comparative Example. The addition of trehalose, on the other hand, does not affect the results.

Experimental Example  1: Cytotoxicity test

In order to verify the safety of the fermented resurrection extract (Example) and the resurrected extract (Comparative Example), each extract was treated with V79-4 cells (Chinese Hamster, continuous cell line of lung fibroblasts) and MTT test Cytotoxicity test was performed by the method. The cytotoxicity test was also performed for the control group, sodium lauryl sulfate (SLS), in the same manner.

As a result, as shown in the following Table 1, the fermented extract of the present invention exhibited cytotoxicity about 900 times lower than that of sodium lauryl sulfate (SLS), which is a surfactant. In addition, it was confirmed that the cytotoxicity was lower than that of the resveratrol extract.

sample IC ₅₀ (% w / v) Example 5.54 Comparative Example 4.23 Sodium lauryl sulfate (SLS) 0.006 [Note] IC ₅₀ (InhibitoryConcentration50): concentration to kill 50% of cells

Experimental Example  2: allergy  Evaluation (H- CLAT )

The H-CLAT (Human Cell Line Activation Test) experiment was conducted to confirm whether the fermented Echinochloa crass extract caused allergic skin.

CV 75 x 1.2, CV 75, CV 75 / 1.2 ³ , CV 75 / 1.2 ⁵ , or a concentration of 10,000 microgram / ml (Highest technical dose) was used as the standard of the cell viability of 75% The fermented transgenic extract (Example) or the transgenic extract (Comparative Example) was treated with THP-1 (human monocytic cell line) and cultured for 24 hours. Then, cell viability was confirmed by MTT assay, and THP-1 cells were stained with fluorescently labeled CD54 and CD86 antibodies, and the amounts of CD54 and CD86 were confirmed by flow cytometry. Thereafter, it was judged to be positive when the CD54 RFI (relative fluorescence intensity) was 200 or more according to the allergenicity criteria, or when the CD86 RFI was 150 or more. The results are shown in Table 2 below.

division Concentration (/ / ml) Cell viability (%) CD54 RFI CD86 RFI Example 4000 86 110 96 5000 87 103 104 8000 91 123 120 100000 89 112 103 Comparative Example 4000 83 110 89 5000 81 120 102 8000 83 113 92 100000 82 128 103

As shown in the table above, according to the allergenicity criteria, it was confirmed that the fermented Echinochloatoid extract had no possibility of causing allergy (negative) at all concentrations exceeding 80% of cell viability.

Experimental Example  3: Evaluation of skin irritation

In order to confirm the irritation of the fermented Echinochloa crus-galli extract to skin, 0.2 g of each sample was patch-tested for 50 minutes in healthy adult male and female subjects, and the pin chamber was removed for 4 hours After the lapse of time, changes in skin condition were visually observed. The results are shown in Table 3 below.

division Number of subjects Judgment result Irritation degree ++ + + - - Negative control group
(Distilled water) 50 - - 2 48 0.07 Positive control group
(SLS 0.1%) 50 4 3 2 41 0.30 Comparative Example 50 - - 2 48 0.10 Example 50 - - 3 47 0.06 [week]
-: No erythema or unusual phenomenon
+ -: slightly reddish from the surroundings
+: Significantly reddened than surrounding
++: More reddening and swelling than the surrounding area

As shown in the above table, the fermented extract of Echinochloa spp. Of the present invention showed low skin irritancy similar to that of the negative control, confirming safety to the skin. The extracts of fermented Echinochloa were lower than those of Echinochloa extract.

Experimental Example  4: whitening effect - melanin total amount reduction effect

In order to confirm the whitening effect of the fermented Echinochloa crus-galli extract, the culture broth of rat melanoma cells (B-16 mouse melanoma cell) was cultured according to the method described in Lotan R. et al. ( Cancer Res . 40: 3345-3350, 1980) , Fermented Echinochloa extract (Experimental Example) was added to measure the total amount of melanin. In the experiment, first the toxicity of melanoma cells in rats was evaluated and the whitening evaluation was carried out at a concentration that is not toxic. DMSO was used as a negative control group, and albutin was used as a positive control group.

Specifically, the material was added to the medium to have a concentration of 100 ppm, and melanoma cells were cultured for 3 days. Then, each cell was treated with trypsin, removed from the culture vessel, and centrifuged. Then, 1 ml of sodium hydroxide solution (1N concentration) was added to dissolve the melanin. Then, the absorbance was measured at 400 nm using a spectrophotometer, and the amount of melanin produced was quantified. The amount of melanin produced (abs) was expressed as the absorbance per unit cell number (1 x 10 ⁶ cells), and the inhibition rate (%) was calculated as the total amount of melanin relative to the negative control. The results are shown in Table 4 below.

sample Melanin production (abs) Inhibition rate (%) The negative control (DMSO) 0.33 - Positive control (arbutin) 0.23 30.3 Comparative Example 0.17 48.48 Example 0.12 63.63

As shown in the table above, the fermented extract of Echinochloa sp. Exhibits a 2-fold inhibitory effect on melanin formation, which is superior to that of arbutin, which acts directly on tyrosinase and inhibits melanin production. Thus, it has excellent whitening effect and inhibits melanin formation It was confirmed that the effect was excellent and the pigmentation was suppressed and the whitening effect was excellent.

Experimental Example  5: Type 1 collagen ( type  I collagen ) Promoting the synthesis of

In order to confirm the effect of improving wrinkles or improving skin elasticity, fermented extracts of Echinochloa or Echinochloa extracts were added to a culture medium of human-derived fibroblasts to examine the promoting effect of collagen synthesis at the cellular level. Measurement of biosynthetic collagen was quantitated using a PICP EIA kit (Procollagen Type I C-Peptide Enzyme Immunoassay KIT, Takara, Cat No. MK10).

First, human normal fibroblasts were cultured in a 5% CO ₂ incubator at 37 ° C for 24 hours in DMEM medium. The fermented extract of Echinochloa or 50 ppm of Echinochloa extract was added to the culture medium and cultured for 48 hours with the control (untreated). Vitamin C was used as a positive control.

After the culture was obtained, the degree of collagen biosynthesis was determined according to each concentration using a PICP EIA Kit, and then measured at 450 nm using a spectrophotometer. The increase rate of collagen production was calculated by the ratio of relative collagen production to the control (untreated) as shown in the following equation, and the results are shown in Table 5 below.

[Equation 1]

Increase in collagen biosynthesis (%) = (amount of test group collagen / amount of control group collagen) x 100

division Sample concentration Increase in collagen biosynthesis (%) Control group (no treatment) 50 ppm - Vitamin C 50 ppm 8 Comparative Example 50 ppm 8 Example 50 ppm 10

As shown in the table, it was confirmed that the fermented extract of Echinochloa sp. Exhibited excellent collagen biosynthesis ability as compared with vitamin C involved in collagen synthesis, and that the collagen synthesis promoting effect was superior to that of the Echinochloa extract.

Experimental Example  6: Collagenase ( Collagenase ) Inhibitory effect

(3- [2-Furyl] acryloyl) -Leu-Gly-Pro-Ala, which is a substrate of collagenase, to inhibit the activity of collagenase, an enzyme that degrades collagen, The buffer (50 mM Tricine, 10 mM CaCl ₂ .H ₂ O, 400 mM NaCl (pH 7.5)) at a concentration of 10 mM, and finally diluted to 75 μM. The collagenase derived from the rat's interest was dissolved in cold 3 rd distilled water at a concentration of 1 unit / ml, diluted with the buffer solution to a final concentration of 0.05 unit / ml, 50 ppm. Epigallocatechin gallate (EGCG) was used as a positive control and DMSO was used as a negative control.

The reaction was carried out in a 96-well plate and allowed to react at room temperature for 10 minutes. Absorbance was measured at 345 nm using a spectrophotometer at intervals of 1 minute, and the maximum slope of absorbance versus time was determined to determine the activity of the enzyme. The inhibition rate of collagenase was Calculated as shown in Equation (2) and shown in Table 6.

&Quot; (2) "

sample Enzyme activity Inhibition rate (%) Control group -0.7 - Positive control group -0.6 14.29 Comparative Example -0.4 42.86 Example -0.3 57.14

As shown in the table, the fermented extract of Echinosophora inhibits the activity of collagenase, an enzyme that degrades collagen, by about 4 times as much as EGCG, which is known to inhibit collagenase, and 1.3 times more It was confirmed that it inhibited. Thus, it was confirmed through Experimental Examples 5 and 6 that the effect of improving the skin wrinkles and skin elasticity of the fermented extract of Echinochloa according to the present invention was improved as compared with that of the Echinochloa extract.

Experimental Example  7: Free radical Scatters  Confirm

The free radical scavenging ability of the fermented resurrection extract was measured by the 1,1-diphenyl-2-picryl hydrazine (DPPH) method (Blois, Nature 181, 1190, 1958). DPPH is a relatively stable free radical, which shows maximum absorption at 517 nm in the presence of radicals and loses its absorbance when radicals are eliminated. DPPH was purchased from Sigma, and dissolved in methyl alcohol at a concentration of 0.15 mM.

First, 100 μl of the fermented resurrection extract or the positive control, vitamin C, was added to each well of a 96-well plate. 100 μl of DPPH solution was added thereto, and the mixture was allowed to stand at room temperature for 30 minutes, and the absorbance at 517 nm was measured using a microplate reader (BioTek EL-340).

The concentration of the extract when the absorbance of the treated sample was half the absorbance of the control group was expressed by IC ₅₀ , and the results are shown in Table 7 below.

sample IC ₅₀ (% w / v) Control group 0.0025 Positive control (vitamin C) concentration 0.0005 Comparative Example Concentration 0.0007 Example  density 0.0006

As shown in the above table, the fermented extract of Echinochloa sp. Showed very strong free radical scavenging ability as compared with vitamin C, and it was confirmed that the extract of Echinochloa sp.

Experimental Example  8: Anti-inflammatory efficacy test

In order to confirm the anti - inflammatory activity of the fermented Echinochloa sp. Extract, the inhibitory activity of NO (NO) was measured by the accumulation amount of NO ₂ .

First, 1 ml of RAW 264.7 cells was added to a 12-well plate so as to be 1 × 10 ⁶ cells / well, and cultured for 24 hours. Thereafter, the medium was replaced with a medium supplemented with LPS (lipopolysaccharide) to induce excessive NO production and a medium not supplemented, and cultured for 1 hour. Then, fermented Echinochloa extract (Example) or Echinochloa extract (Comparative Example) was treated And cultured for 24 hours.

After separation of the supernatant of 100μl in each well, Greece (Griess) solution of 100μl in a blank well (dissolved in distilled water to 0.1% NED (naphthyl ethylene diamine is the hydrochloride) and dissolved in a 5% H ₃ PO ₄ 1: 1 mixture of 1% sulfanilamide) and reacted in the dark. Then, the absorbance at 550 nm was measured using a molecular device microplate reader (Sunnyvale, Calif., USA) to measure the amount of nitrogen dioxide accumulated in the medium.

sample NO production (μM) Control group 1.5 LPS treatment (positive control) 9.8 LPS Treatment + Comparative Example 8 LPS treatment + Example 7.5

As a result, as shown in Table 8, when the LPS treatment was performed, the NO production was increased compared to the control (no treatment), but it was confirmed that the treatment of the fermented extract of Echinochloa can effectively inhibit NO production despite LPS treatment . In particular, it was confirmed that the NO inhibiting activity was better in the experimental example than the comparative example in which the extract was treated. Thus, the fermented extract of Echinochloa can alleviate the inflammation caused by an increase in NO production, an inflammatory mediator.

Manufacturing example  1, 2, compare Manufacturing example  1, 2: Preparation of essence

The extracts of Experimental Examples 1 and 2 were prepared in the same manner as in Example 1 except that the extracts of Comparative Examples and Examples were mixed at a weight ratio of 1: 1. In addition, an essence containing Comparative Example 1 (extract of Echinochloa sp.) Was prepared in Comparative Preparation Example 1, and an essence containing no extract of Experimental Example or Comparative Example was prepared in Comparative Preparation Example 2. The detailed composition (% by weight) is shown in Table 9 below.

Category (% by weight) Production Example 1 Production Example 2 Comparative Preparation Example 1 Comparative Production Example 2 Purified water Balance Balance Balance Balance Triethanolamine 0.25 0.25 0.25 0.25 Carboxyvinyl polymer ¹ 0.22 0.22 0.22 0.22 glycerin 4 4 4 4 Propylene glycol 0.2 0.2 0.2 0.2 Paraoxybenzoic acid ester 0.2 0.2 0.2 0.2 Comparative Example - - 7 - Example 7 - - - A 1: 1 mixture of Comparative Example 2 - 7 - - Wax 0.5 0.5 0.5 0.5 Cetostearyl alcohol One One One One Glyceryl monostearate One One One One Self-emulsifying type
Monostearic acid One One One One Sorbitan monostearate ² 0.5 0.5 0.5 0.5 Polyethylene glycol monostearate 8 8 8 8 P-hydroxybenzoic acid profile 0.1 0.1 0.1 0.1 Dimethylsiloxane 0.3 0.3 0.3 0.3 Isocetyl octanoate ³ 3 3 3 3 Squalane 5 5 5 5 Spices 0.05 0.05 0.05 0.05 total 100 100 100 100

[week]

1. Carboxyvinyl polymer - US Ratio. F. A 1 wt% aqueous solution of Carbopol 941 from B.F. Goodrich Co.

2. Sorbitan monostearate - ICI Sialasel 60 (Arlacel 60)

3. Isocetyl octanoate - Ishihichi (ICEH) from Nihon KoKyu Alcohol Co.,

Experimental Example  9: Moisture Possession  evaluation

In order to evaluate the skin moisturizing power of the prepared essence, each essence was applied to the skin in a predetermined amount, and then the moisture content of the stratum corneum was measured and evaluated. As a measuring device, a skin moisture meter (skin surface hygrometer, model name: SKICON-200, manufactured by IBS Japan) was used.

Specifically, a certain amount (2 mg / cm 2) of each essence was applied to the inside of each arm of 20 subjects in a constant temperature and humidity room at 25 ° C. and 45% relative humidity, and rubbed thoroughly. And the water increase rate (%) after 3 hours were measured to evaluate the moisturizing effect. The results obtained are shown in Table 10 below.

division Moisture growth rate (%) Production Example 1 Production Example 2 Comparative Preparation Example 1 Comparative Production Example 2 Before application 0 0 0 0 After 1 hour of application 57 51 45 23 After 3 hours of application 39 31 26 15

As shown in the above table, the essence of Preparation Example 1 containing the fermented extract of Echinochloa according to the present invention contains not only Comparative Preparation Example 2 which does not contain the extract but also essence of Comparative Preparation Example 1 which contains the fermented Echinochloa extract The water retention increase rate was improved after one hour and three hours after the application, and it was confirmed that the water retention showed excellent moisturizing effect.

Manufacturing example  3, 4, compare Manufacturing example  3, 4: Underwater type Emulsion  Produce

An aqueous type emulsion containing a mixture obtained by mixing the extract of the above Experimental Examples or the extracts of Comparative Examples and Examples at a weight ratio of 1: 1 was prepared as Preparation Examples 3 and 4. [ In addition, an underwater type emulsion containing Comparative Example 1 and an extract of Experimental Example or Comparative Example was prepared in Comparative Preparation Example 2, except for the comparative example (resurrection extract). The detailed composition (% by weight) is shown in Table 11 below.

Category (% by weight) Production Example 3 Production Example 4 Comparative Production Example 3 Comparative Production Example 4 Awards Purified water Balance Balance Balance Balance Triethanolamine 0.2 0.2 0.2 0.2 Carboxyvinyl polymer ¹ 15 15 15 15 glycerin 5 5 5 5 Propylene glycol 5 5 5 5 Paraoxybenzoic acid ester 0.2 0.2 0.2 0.2 Example - - 0.1 - Example 2 0.1 - - - 1: 1 mixture of Examples 1 and 2 - 0.1 - - Cetostearyl alcohol One One One One Glyceryl monostearate One One One One Sorbitan monostearate ² One One One One POE (20) sorbitan monostearate 1.5 1.5 1.5 1.5 P-hydroxybenzoic acid profile 0.1 0.1 0.1 0.1 Isocetyl myristate 5 5 5 5 Isocetyl octanoate ³ 5 5 5 5 Squalane 5 5 5 5 Spices 0.05 0.05 0.05 0.05 100 100 100 100 Isocetyl octanoate ³ 5 5 5 5 Squalane 5 5 5 5 Spices 0.05 0.05 0.05 0.05 total 100 100 100 100 [week]
1. Carboxyvinyl polymer - US Ratio. F. A 1% by weight aqueous solution of Carbopol 941 from BF Goodrich Co. 2. Sorbitan monostearate - ICI Sialasel 60 (Arlacel 60) 3. Isocetyl octanoate - Ishihichi (ICEH) from Nihon KoKyu Alcohol Co.,

Experimental Example  10: Wrinkle improvement effect

The effect of improving the wrinkles of the emulsions of the underwater type was measured by the following method in healthy 35-50 year old women.

80 women aged 35 to 50 years were divided into 8 groups by 10 persons. Seven groups were applied to the facial region of the underwater type emulsion of Production Example 3, 4, or Comparative Production Example 3 once a day for 3 months, The remaining one group was applied to the facial part of the Comparative Example 4 for 3 months once a day.

After 3 months, the improvement of wrinkles was evaluated by image analysis of questionnaires and wrinkles. The questionnaire of the subject was judged to have no improvement, slight improvement, and considerable improvement as compared with before use with respect to wrinkle improvement and elasticity improvement, and the results are shown in Table 12 below.

division No improvement Slight improvement A significant improvement Manufacturing example  3 One 5 4 Production Example 4 One 7 2 Comparative Production Example 3 One 6 3 Comparative Production Example 4 7 3 0

As shown in the above table, the essence of Preparation Example 3 containing the fermented extract of Echinochloa according to the present invention contains not only Comparative Preparation Example 4 which does not contain the extract but also essence of Comparative Preparation Example 3 which contains an unfermented Echinochloa extract It was confirmed that the effect of improving wrinkles was excellent.

Formulation example  One: Cosmetic  Produce

1. Manufacture of lotion

0.1% by weight of glycerin, 3.0% by weight of dipropylene glycol, 0.5% by weight of hyaluronic acid, 0.1% by weight of polyoxyethylene hydrogenated castor oil, 0.1% by weight of polyethylene oxide oleate (Total composition: 100% by weight), 5.0% by weight of ethanol, 0.15% by weight of preservative, 0.05% by weight of pigment, 0.05% by weight of perfume and remaining balance.

2. Manufacture of Essence

7.0% by weight of catecholamines, 1.0% by weight of cetostearyl alcohol, 1.0% by weight of self-emulsifiable monostearate, 0.5% by weight of waxes, 5.0% by weight of squalane, 3.0% by weight of isocetyl octanoate 0.3 wt% of dimethylsiloxane, 0.5 wt% of monostearic acid sorbitan, 8.0 wt% of monostearic acid polyethylene glycol, 4.0 wt% of glycerin, 0.2 wt% of propylene glycol, 0.22 wt% of carboxy polymer, 0.25 wt% of triethanolamine, 0.05 wt% %, 0.05% by weight of coloring matter, 0.05% by weight of fragrance, and the remaining amount of purified water (total composition: 100% by weight).

3. Production of Lotion

5.0% by weight of cetyl alcohol, 0.8% by weight of cetostearyl alcohol, 1.0% by weight of self-emulsifying monostearate, 0.5% by weight of wax, 0.5% by weight of stearic acid, 7.0% by weight of liquid paraffin, 5.0% by weight, macadamia oil 3.0% by weight, isocetyl octanoate 2.0% by weight, dimethylsiloxane 0.3% by weight, monostearic acid sorbitan 0.5% by weight, monostearic acid polyethylene glycol 1.2% by weight, glycerin 4.0% by weight, propylene glycol (100% by weight of total composition) of 4.0% by weight, betaine 4.0% by weight, carboxypolymer 0.12% by weight, triethanolamine 0.15% by weight, preservative 0.25% by weight, coloring matter 0.05% by weight, To prepare a lotion.

4. Manufacture of cream

7.0% by weight of cetostearyl alcohol, 0.5% by weight of self-emulsifiable monostearic acid, 1.5% by weight of pro-type monostearic acid, 0.5% by weight of wax, 8.0% by weight of liquid paraffin 4.0% by weight of squalane, 4.0% by weight of isocetyl octanoate, 4.0% by weight of purified jojoba oil, 0.3% by weight of dimethylsiloxane, 1.0% by weight of monostearic acid sorbitan, 1.2% by weight of monostearic acid polyethylene glycol, (Whole composition: 100 wt%) was mixed with 4.0 wt% of propylene glycol, 4.0 wt% of betaine, 0.06 wt% of xanthan gum, 0.10 wt% of triethanolamine, 0.25 wt% of preservative, 0.05 wt% ) To prepare a cream.

Formulation example  2: Preparation of pharmaceutical composition

1. Preparation of gel

1.0% by weight of glycerin, 4.0% by weight of propylene glycol, 10% by weight of ethanol, 0.1% by weight of polyoxyethylene hydrogenated castor oil, 0.30% by weight of carboxy polymer (Total composition: 100% by weight), 0.30% by weight of triethanolamine, 0.05% by weight of preservative, 0.05% by weight of pigment, 0.05% by weight of fragrance and a remaining amount of purified water.

2. Manufacture of ointment

0.5% by weight of beta-1, 3-glucan, 10.0% by weight of beeswax, 5.0% by weight of Polysorbate 60, 60% by weight of fermented castor oil extract 2.0 weight%, sorbitan sesquioleate 0.5 weight%, vaseline 5.0 weight%, liquid paraffin 10.0 weight%, squalane 5.0 weight%, shea butter 3.0 weight%, caprylic / capric triglyceride 5.0 weight%, glycerin 10.0 weight (Whole composition: 100% by weight), 10% by weight of propylene glycol, 0.2% by weight of triethanolamine, 0.05% by weight of preservative, 0.05% by weight of pigment, 0.05% by weight of perfume and 100% by weight of purified water.

3. Preparation of topical medicament (patch)

0.5% by weight of beta-1,3-glucan, 20.0% by weight of hexylene glycol, 0.7% by weight of diethylamine, 1.0% by weight of polyacrylic acid (Carbopol 934P) , 0.1% by weight of sodium sulfite, 1.0% by weight of polyoxyethylene lauryl ether (EO = 9), 1.0% by weight of polyhydroxyethylene cetyl stearyl ether (Cetomacrogol 1000), 2.5% by weight of viscous paraffin oil / Capric acid ester (Cetiol LC), 3.0% by weight of polyethylene glycol 400, and the remaining deionized water (100% by weight of the whole composition).

From the above description, it will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention without departing from the scope of the present invention as defined by the appended claims.

Claims (11)

A cosmetic composition for skin whitening, preventing or improving skin wrinkles, improving skin elasticity, or antioxidation, comprising fermented extract of Echinochloa as an active ingredient.
The method according to claim 1,
Wherein the composition further comprises an extract of Echinochloa sp. As an active ingredient.
The method according to claim 1,
Wherein the fermented extract of Eucalyptus is an extract of Eucalyptus fermented by at least one selected from the group consisting of mushroom, lactic acid bacteria and yeast.
The method of claim 3, wherein
Wherein the yeast is Saccharomyces cerevisiae .
The method of claim 3,
Wherein the fermentation is carried out at 10 占 폚 to 35 占 폚 for 24 hours to 5 days.
The method according to claim 1,
Wherein the fermented extract of Eucalyptus is water and ethanol extract of fermented Eucalyptus.
3. The method according to claim 1 or 2,
Wherein the active ingredient is contained in an amount of 0.0001% by weight to 30% by weight based on the total weight of the total composition.
Wherein the active ingredient is at least one selected from the group consisting of fermented extracts of Echinochloa crus-galli and extracts of Echinochloa.
9. The method of claim 8,
Wherein the fermented extract of Eucalyptus is an extract of Eucalyptus fermented by at least one selected from the group consisting of mushroom, lactic acid bacteria and yeast.
9. The method of claim 8,
Wherein the composition is an external preparation for skin.
A pharmaceutical composition for antioxidation, which comprises fermented extract of Echinochloa as an active ingredient.