KR101125225B1 - A composition for skin wrinkle improvement comprising extracts or fractions of Eremochloa ophiuroides as an active ingredient - Google Patents

Abstract

The present invention is sentigrass ( Eremochloa) ophiuroides ) extract, fractions thereof, or a composition for improving skin wrinkles containing the active fractions isolated from the fractions as an active ingredient, more specifically centifedgrass extract, fractions thereof or the active fractions separated from the fractions are elastin Since it has an excellent inhibitory activity of elastase that degrades it, it can be usefully used in a composition for improving skin wrinkles.

Sentipedgrass (Eremochloa ophiuroides), elastase, wrinkle improvement

Description

A composition for skin wrinkle improvement comprising extracts or fractions of Eremochloa ophiuroides as an active ingredient}

The present invention relates to a composition for improving skin wrinkles containing centipede glass extract or a fraction thereof as an active ingredient.

The main mechanism involved in the formation of skin wrinkles is that the skin is continuously exposed to oxidative stress from harmful environment such as air pollution, UV exposure, stress or disease, increasing the radicals in the body, and collagen (collagen), which is the connective tissue of the dermis. ), Elastin, hyaluronic acid (hyaluronic aicd) can be destroyed by causing a certain part of the skin subsidence (wrinkling) is described. In addition, these mechanisms can oxidize the lipid portion of the cell membrane and cause cell destruction, leading to diseases such as dermatitis, acne or skin cancer. In addition, radicals are involved in the process of melanin formation, causing spots, freckles and wrinkles.

To improve skin wrinkles, ascorbic acid, α-tocopherol, retinol and derivatives thereof, and superoxide dismutase (SOD) have been used in cosmetics or pharmaceuticals as substances for improving free radical scavenging or skin wrinkles. Although it has been used in combination to prevent wrinkles and other skin diseases, they are not only expensive, but also have poor chemical stability, which makes it difficult to expect a substantial effect. For this reason, the development of a safer and more effective material for improving skin wrinkles has emerged as an important task not only in the pharmaceutical and food sectors but also in the cosmetics industry.

Retinoids used in skin antiwrinkle cosmetics include retinol, retinal, retinoic acid, retinyl acetate, retinyl linoleate and retinyl palmitate ( retinyl palmitate). Among these, retinol is an endogenous compound that is naturally present in the human body and is essential for the differentiation and growth of epithelial tissues.It has very high safety compared to retinoids such as retinic acid, and thus is suitable as a raw material for skin wrinkle improvement cosmetics. .

These retinoids have been proven to be effective in improving wrinkles on the skin, but when prescribed in the product, discoloration and odor occur, and skin irritation occurs even when used in small amounts. The reduction of the effect is recognized as a big problem.

In order to overcome the problems of the retinoids, it is necessary to develop a cosmetic for improving wrinkles using natural-derived bioactive substances. This has the advantage of less skin side effects, less chemical changes over time compared to retinoids, excellent safety and stability of cosmetics. Therefore, research on medicinal plant resources as a cosmetic for skin wrinkle improvement continues.

Sentipedgrass ( Eremochloa ophiuroides ) is a slow growing turbulent turf, a perennial grass that crawls and forms turf. It belongs to the Poaceae family and its scientific name is Eremochloa ophiuroides (Munro) Hack. It is mainly distributed in China, East Asia, Indochina, the northeastern United States, Meso America, and the Caribbean, and is widely adapted to various soil conditions, but is especially resistant to moist, low acidic soils and low-quality sandy soils.

As a morphological feature, the leaves are 15-30 mm long, 2-4 mm wide, flat with a mid vein, hairless except for the collar part, and the leaf ends are round. The stem has a very firm sheath, and the roots are thin branched stolons. The flower part has one raceme of about 3 to 5 inches, which has a purple, slightly flat, small spikelet arranged in two rows. Have Its main use is as a lawn or turf grass and as a forage due to its high leafiness and good taste.

Therefore, the present inventors have studied to develop a natural plant extract for skin wrinkle improvement, the skin extract, the extract extracted from Sentipedgrass, its fractions and the active fractions separated from the fractions have an elastase inhibitory activity The present invention has been completed by revealing that it can be usefully used as an active ingredient of the composition for improvement.

The object of the present invention is Centipedgrass ( Eremochloa) It is to provide a composition for improving skin wrinkles containing ophiuroides ) extract or a fraction thereof as an active ingredient.

In order to achieve the above object, the present invention provides a cosmetic composition for improving skin wrinkles containing centipede glass extract, a fraction thereof or an active fraction separated from the fraction as an active ingredient.

In another aspect, the present invention provides a pharmaceutical composition for skin wrinkle improvement containing the centipedes grass extract, fractions thereof or active fractions isolated from the fractions as an active ingredient.

In addition, the present invention provides a skin wrinkle improvement health food containing the centifed glass extract, a fraction thereof or an active fraction separated from the fraction as an active ingredient.

Centipedgrass extract, fractions thereof or active fractions isolated from the fractions can be usefully used as a composition for improving skin wrinkles because they have an activity of inhibiting elastase that destroys elastin.

Hereinafter, the present invention will be described in detail.

The present invention provides a cosmetic composition for improving skin wrinkles containing centipede glass extract, a fraction thereof or an active fraction separated from the fraction as an active ingredient.

The centipedes grass extract is preferably prepared by a manufacturing method comprising the following steps, but not always limited thereto:

1) extracting by adding an extracting solvent to centipede glass;

2) cooling the extract of step 1) and filtering; And

3) concentrating the filtered extract of step 2) under reduced pressure.

In the above method, the centipedes of step 1) can be used without limitation, such as those grown or commercially available. The centifed grass is preferably used as a leaf, but is not limited thereto.

The extraction solvent is preferably water, alcohol or a mixture thereof. As the alcohol, C ₁ to C ₄ lower alcohols are preferably used, and as the lower alcohols, ethanol or methanol is preferably used. As the extraction method, it is preferable to use shaking extraction or reflux extraction, but not always limited thereto. The extraction solvent is preferably extracted by adding 2 to 10 times the amount to the centipedes glass, but is not limited thereto. Extraction temperature is preferably 50 to 100 ℃ but is not limited thereto. In addition, the extraction time is preferably 2 to 5 days, but is not limited thereto. In addition, the extraction number is preferably one to three times, but is not limited thereto.

In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in step 3), but it is not limited thereto. In addition, the drying is preferably reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, but is not limited thereto.

The fraction of the centifed glass extract or the active fraction separated from the fraction is preferably prepared by a manufacturing method comprising the following steps, but not always limited thereto:

1) adding an organic solvent to the centipedes glass extract to prepare an organic solvent fraction; And

2) preparing an active fraction by subjecting the fraction of step 1) to column chromatography.

In the above method, the organic solvent of step 1) is preferably ethyl acetate, but not always limited thereto.

In the above method, the column chromatography of step 2) is an active fraction by performing column chromatography using a filler selected from the group consisting of silica gel, Sephadex, RP-18, polyamide, Toyopearl and XAD resin. Can be isolated and purified. Column chromatography can be carried out several times by selecting an appropriate filler as necessary.

We adsorbed the ethyl acetate fractions of centifedgrass to Flex-Colum ^™ (3 cm × 25 cm) and Toyoperarl HW-40 (75 μm) column chromatography and dissolved 20, 50 or 70% methanol. Using ethyl acetate-20% methanol, ethyl acetate-50% methanol and ethyl acetate-70% methanol were obtained.

The centipedes grass extract preferably includes any one or more of compounds selected from the group consisting of compounds represented by the following Chemical Formulas 1 to 8, but is not limited thereto.

The active fraction separated from the fraction of the centipedes grass extract preferably includes any one or more of compounds selected from the group consisting of compounds represented by the following formulas (4) to (8) (but not limited thereto) , And FIG. 5).

In order to investigate the skin wrinkle improvement effect of centifed glass extract, the present inventors found that elastase inhibitory activity according to the concentration of centifed glass extract (20, 200, 2000 or 4000 ㎍ / ml) was applied to the skin wrinkle improvement activity. It was measured in comparison with the well-known corn beard extract which was known to be excellent. As a result, the elastase inhibitory activity of the centipedes grass extract at concentrations of 20 to 4000 μg / ml was about 24, 50, 91 and 93% for methanol extracts and about 23, 50, 90 and 92% for ethanol extracts, respectively. , Water extracts of about 23, 49, 90 and 91%, respectively, and the corncoal methanol extract showed about 8, 17, 59 and 75% of elastase inhibitory activity at the same concentrations (Table 2 and 6). Therefore, it can be seen that the centipedes grass extract has a significantly better skin wrinkle improvement effect than the corn beard extract.

The inventors of the present invention, in order to determine the skin wrinkle improvement effect of the centifed glass extract, fractions thereof or the active fractions isolated from the fractions, the ella of 4000 ㎍ / ㎖ Centifgrass grass extracts, fractions or active fractions separated from the fractions Statase inhibitory activity was measured. As a result, the centipedes grass extract showed about 96-98% elastase inhibitory activity, and the active fraction of the ethyl acetate fraction showed about 92% elastase inhibitory activity (see Table 3 and FIG. 7).

Therefore, the centipedes grass extract, fractions thereof or the active fractions separated from the fractions have a significant anti-wrinkle effect, and thus may be usefully used as an effective cosmetic composition for skin wrinkle improvement.

The cosmetic composition is, for example, emulsions, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes), nonionic vesicle dispersants obtained by dispersing an oil phase in a solution, a gel, a solid or pasty anhydrous product, or an aqueous phase. It may be provided in the form of creams, skins, lotions, powders, ointments, sprays or concealed sticks. It may also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

In addition, the cosmetic composition, in addition to the centipedes grass extract of the present invention, fractions thereof or the active fraction separated from the fractions, fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers Agents, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic Or adjuvants commonly used in the cosmetic field, such as lipophilic actives, lipid vesicles, or any other ingredient conventionally used in cosmetics.

In the cosmetic composition containing the centipede grass extract of the present invention, a fraction thereof or an active fraction separated from the fraction, in the cosmetic composition usually contained 1 to 15% by weight, preferably 2 to 10 It can be added in an amount by weight.

In the case of using the centipedes grass extract of the present invention, fractions thereof, or active fractions separated from the fractions as external preparations for skin, additionally, fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, Stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments , Hydrophilic or lipophilic actives, lipid vesicles, or any other ingredient commonly used in external preparations for skin, may contain adjuvants commonly used in the field of dermatology. In addition, the ingredients may be introduced in amounts generally used in the field of dermatology.

     In another aspect, the present invention provides a pharmaceutical composition for skin wrinkle improvement containing the centipedes grass extract, fractions thereof or active fractions isolated from the fractions as an active ingredient.

Centifedgrass extract, fractions thereof, or active fractions isolated from the fractions of the present invention were found to have elastase inhibitory activity and were found to have excellent skin anti-wrinkle effect, which is useful as an effective pharmaceutical composition for skin anti-wrinkle improvement. Can be used.

The pharmaceutical composition of the present invention may further include conventionally used excipients, disintegrants, sweeteners, lubricants, flavoring agents, etc., tablets, capsules, powders, granules, suspensions, emulsions, It can be formulated into syrups, other liquids.

In particular, the pharmaceutical compositions of the present invention may be formulated for oral administration such as, for example, stagnation, troches, lozenges, aqueous or dominant suspensions, prepared powders or granules, emulsions, hard or soft capsules, syrups or It is formulated with elixirs. Lactose, saccharose, sorbitol, mannitol, starch, amylopectin, binders such as cellulose or gelatin, excipients such as dicalcium phosphate, disintegrants such as corn starch or sweet potato starch, for formulation into formulations such as tablets and capsules, Lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax. In the case of a capsule formulation, in addition to the above-mentioned substances, a liquid carrier such as fatty oil is contained.

In addition, the pharmaceutical composition of the present invention may be administered orally or parenterally, and it is preferable to select subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection injection method. To formulate into parenteral dosage forms, the centifedgrass extract, fractions or active fractions thereof of the present invention are mixed in water with stabilizers or buffers to prepare a suspension and formulated in unit dosage forms of ampoules or vials.

The dosage of the active ingredient according to the present invention is appropriately selected according to the absorption rate, inactivation rate and excretion rate of the active ingredient in the body, the age, sex and condition of the patient, the severity of the disease to be treated, but in the case of oral administration in general Adults can be administered once to several times in an amount of 0.0001 to 500 mg of the extract of the present invention per kg of body weight per day, it is preferable to administer in an amount of 0.001 to 100 mg.

In addition, the present invention provides a skin wrinkle improvement health food containing the centifed glass extract, a fraction thereof or an active fraction separated from the fraction as an active ingredient.

Centifed glass extract, fractions thereof, or active fractions separated from the fractions of the present invention have elastase inhibitory activity and thus can be usefully used as an effective food for skin wrinkle improvement.

The health food of the present invention may be added to the centipedes grass extract, fractions thereof, or active fractions separated from the fractions as it is, or used with other foods or food ingredients, and may be appropriately used according to conventional methods.

There is no particular limitation on the kind of food. Examples of foods to which the centifed grass extract, fractions thereof, or active fractions separated from the fractions may be added include meat, sausage, bread, chocolate, candy, snacks, confectionary, pizza, ramen, other noodles, gum, and ice cream. Dairy products, including various soups, beverages, tea, drinks, alcoholic beverages and vitamin complexes, and the like, includes all of the health food in the usual sense.

The health beverage composition of the present invention may contain various flavors, natural carbohydrates, and the like as additional components, as in a conventional beverage. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.

In addition to the above, the centifedgrass extract, fractions thereof, or active fractions separated from the fractions of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and salts thereof, organic acids, protective colloids. Thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks and the like. In addition, the centifedgrass extract, fractions thereof or active fractions of the present invention may contain pulp for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not critical but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the present invention will be described in detail by Examples, Experimental Examples, Comparative Examples and Preparation Examples.

However, the following Examples, Experimental Examples, Comparative Examples, and Production Examples specifically illustrate the present invention, and the content of the present invention is not limited to Examples, Experimental Examples, Comparative Examples, and Production Examples.

< Example  1> Sentipedgrass  Preparation of Extract

<1-1> Sentipedgrass  Preparation of Methanol Extracts

Sentipedgrass seeds from Fukukaen Nursery and Blub co. It was imported from the Korea Atomic Energy Research Institute, and was grown on the pavement of Jeongeup Radiation Science Research Institute. The collected leaf samples were stored and used at -80 ° C. 5 kg of centipedgrass leaves were added to 20 L of 100% methanol (MeOH) and crushed in a blender, and then the extract obtained by immersion at room temperature for 3 days was concentrated under reduced pressure at 50 ° C. to obtain 210 g of centipedes methanol extract.

<1-2> Sentipedgrass  Preparation of Ethanol Extract

Sentipedgrass seeds from Fukukaen Nursery and Blub co. Ltd. was imported from the Korea Atomic Energy Research Institute Chung-eup Institute of Radiation Science Research Institute and collected from the Centipedgrass leaf. The collected leaf samples were stored and used at -80 ° C. 5 kg of centipedgrass leaves were added to 20 L of 100% ethanol (EtOH) and crushed with a blender, and then the extract obtained by immersion at room temperature for 3 days was concentrated under reduced pressure at 50 ° C. to obtain 203 g of centipedes ethanol extract.

<1-3> Sentipedgrass  Preparation of Water Extract

Sentipedgrass seeds from Fukukaen Nursery and Blub co. Ltd. was imported from the Korea Atomic Energy Research Institute Chung-eup Institute of Radiation Science Research Institute and collected from the Centipedgrass leaf. The collected leaf samples were stored and used at -80 ° C. 5 kg of centipedgrass leaves were added to 20 liters of water, and crushed by a blender. The extract obtained by immersion at room temperature for 3 days was concentrated under reduced pressure at 50 ° C. to obtain 198 g of centipedes glass water extract.

< Example  2> Sentipedgrass Fraction  And preparation of active fractions

The centifed glass methanol extract obtained in Example 1-1 was fractionated according to polarity using an organic solvent. As shown in FIG. 1, the concentrated methanol extract was suspended in water, first extracted with hexane (N-Hexane), which is a low polar solvent, and the water layer was partitioned with ethyl acetate (EtOAc). After partitioning into two solutions, a soluble fraction with water was finally obtained. The solvent extract fractions were concentrated under reduced pressure and dried to obtain an ethyl acetate fraction (21 g).

20%, 50% or 70% of ethyl acetate fraction was used as the eluent and Flex-Colum ^TM (3 cm × 25 cm) and Column chromatography was performed using Toyoperarl HW-40 (75 μm), ethyl acetate-20% methanol (9.41 g), ethyl acetate-50% methanol (5.74 g) and ethyl acetate-70% methanol (1 g). Three active fractions were obtained.

< Comparative example  1> Preparation of Corn Beard Extract

Corn beard samples were collected from a farm of Kwangpyeongok varieties from farms in Jecheon, Chungbuk. 5 kg of corn beard was added to 20 L of 80% methanol (MeOH), and then pulverized with a blender. The extract obtained by immersion at room temperature for 3 days was concentrated under reduced pressure at 50 ° C. to obtain 194 g of corn beard methanol extract.

< Experimental Example  1> Sentipedgrass  Extract or its From fractions  Useful secondary present in isolated active fraction Metabolite  analysis

Centipedgrass methanol extract using Agilent high performance liquid chromatography (Agilent Technological, Palo Alto, Calif., USA), ethyl acetate-20% methanol, ethyl acetate-50 isolated from the extract Useful secondary metabolites of% methanol and ethyl acetate-70% methanol active fractions were analyzed. The column used Beckman Coulter ULTRASPHERE (5 μm, 4.6 mm × 25 cm) and the temperature of the column was adjusted to 40 ° C. HPLC was carried out using a mobile phase containing a solution containing 0.1% trifluoroacetic acid (A) and a 50% acetonitrile solution (B) containing 0.1% trifluoroacetic acid as a mobile phase. . According to the mobile phase conditions shown in Table 1 below, the flow rate was maintained at 1 ml / min for 50 minutes.

As a result, the centifedgrass methanol extract contains useful secondary metabolites such as chlorogenic acid, rutin (Quercetin-3-O-rutinoside), and naringinin. Naringin (naringenin-7-O-neohesperidoside), isoorientin, rhamnosylisoorientin, iso-ipoline (apigenin-7-O) -Isotinifol {isorhoifolin (apigenin-7-O-rutinoside)}, isochaptoside (apigenin-6- C- arabinosyl-8-C-glucoside {isoschaftoside (Apigenin-6- C- arabinosyl-8) C- glucoside) and maysiin (FIG. 2).

Secondary metabolites such as isoorientin, rhamnosylisoorientin, and isoleuropoline (apigenin-7-O-rutinoside) are also useful for centrifuged glass ethyl acetate-20% methanol active fractions {isorhoifolin (apigenin-7-O-rutinoside)}, isochaptoside (apigenin-6- C- arabinosyl-8-C-glucoside {isoschaftoside (Apigenin-6- C- arabinosyl-8- C -glucoside ) And maysiin (Fig. 3) .The active metabolite isoorientin, rhamnosylisoorientin, useful for centifedgrass ethyl acetate-50% methanol active fractions. ), Isoleuropoline (apigenin-7-O-rutinoside) {isorhoifolin (apigenin-7-O-rutinoside)} and maysiin were shown to exist (FIG. 4). The acetate-70% methanol active fraction contains maysiin, a useful secondary metabolite. It appeared to exist (FIG. 5).

Mobile phase A: 0.1% Trifluoroacetic acid (TFA) B: Acetonitrile: Water (H ₂ O) = 1: 1 (v / v) in 0.1% TFA Extraction mode
(Gradient conditions) Extraction time % Mobile phase B 0 20 18 33 30 40 40 40 42 20 50 20

< Experimental Example  2> elastase ( elastase ) Measurement of wrinkle improvement effect through inhibitory activity

<2-1> Sentipedgrass  Wrinkle improvement effect according to the concentration of extract

Measurement of pancreatic elastase inhibitory activity in pigs was carried out using N -succinyl- (L-alanine) ₃ - p -nitroanilide { N -succinyl- (L-Ala) ₃ - p -nitroanilide} as substrate. The production amount of p -nitroanilide for 30 minutes at 37 ° C was measured at 405 nm. Elasta dissolved in 50 mM Tris-HCl buffer solution (Tri-HCl buffer (pH 8.6)} in 100 μl of the centipedes grass extract obtained in Example 1 or the corn beard extract obtained in Comparative Example 1. 100 [mu] l of a 2.5 unit solution was added followed by 100 [mu] l of 0.4 M Tris-HCl (Tri-HCl, pH 8.6). After addition, substrate N -succinyl- (L-alanine) ₃ - p -nitroanilide { N -dissolved in 200 μl of substrate solution [50 mM Tris-HCl buffer (pH 8.6)}. succinyl- (L-Ala) ₃ - p -nitroanilide (0.5 mg / ml)} to a total volume of 500 μl and the final concentration of the extract to 20, 200, 2000 or 4000 μg / ml, 37 ° C. After reacting for 20 minutes at, it was measured at 405 nm. 100 ㎍ / ㎖ assolic acid (ursolic acid) was used as a positive control, the negative control was measured using a solvent dissolved sample. Elastase inhibitory activity is expressed as the absorbance decrease rate of the addition group and no addition group of the extract, it was calculated according to the following [Equation 1].

% Inhibition = (absorbance of 1-sample addition group / absorbance of no addition group) × 100

As a result, the antiwrinkle effect of the centipedes glass methanol extract showed about 24, 50, 91 and 93% of elastase inhibitory activity at concentrations of 20, 200, 2000 and 4000 ㎍ / ml, respectively. Showed elastase inhibitory activity of about 8, 17, 59 and 75% at concentrations of 20, 200, 2000 and 4000 μg / ml, respectively (Table 2 and FIG. 6). Therefore, it was confirmed that the centipedes grass extract has a significantly higher skin wrinkle improvement effect than the corn beard extract.

sample Final concentration (㎍ / ㎖) Elastase Inhibition Rate (%) sen
tea
Fe
De
That
la
S





Methanol extract


20 24 200 50 2000 91 4000 93 Ethanol extract


20 23 200 50 2000 90 4000 92 Water extract


20 23 200 49 2000 90 4000 91 corn
mustache Methanol extract 20 8 200 17 2000 59 4000 75 Asolic acid 100 98

<2-2> Sentipedgrass  Extract, its Fraction  Or above From fractions  Determination of Antioxidant Activity from Isolated Active Fractions

In accordance with the method of Experimental Example <2-1>, the elastase inhibitory activity of the 4000 μg / ㎖ Centifed Grass extract, fractions thereof, or the active fractions isolated from the fractions was measured.

As a result, centifedgrass methanol, ethanol or water extract showed about 98, 96 and 97% elastase inhibitory activity, and the active fraction isolated from the ethyl acetate fraction showed about 92% elastase inhibitory activity. (Table 3 and FIG. 7).

Sample (4000 μg / ml) Elastase Inhibition Rate (%) Methanol extract 98 Ethanol extract 96 Water extract 97 Ethyl acetate-20% methanol 92 Ethyl acetate-50% methanol 92 Ethyl Acetate-70% Methanol 92

< Manufacturing example  1> Sentipedgrass  Extract, its Fraction  Or above From fractions  For wrinkle improvement of skin containing separated active fraction as active ingredient Cosmetic  Produce

<1-1> preparation of flexible lotion

The flexible purified water containing the centipede glass ethanol extract of Example <1-2> as an active ingredient was prepared as shown in [Table 4].

Raw material Content (parts by weight) Ethanol Extract of Example <1-2> 10.00 1,3-butylene glycol 1.00 Disodium ID 0.05 Allantoin 0.10 Dipotassium glycylizate 0.05 Citrix Acid 0.01 Sodium citrate 0.02 Glyceres-26 1.00 Arbutin 2.00 Hydrogenated Castor Oil 1.00 ethanol 30.00 Preservative a very small amount coloring agent a very small amount Flavoring agent a very small amount Purified water Balance

<1-2> Preparation of nourishing cream

Nutrients containing the centipede glass ethanol extract of Example <1-2> as an active ingredient was prepared as shown in the composition of the following [Table 5].

Raw material Content (parts by weight) Ethanol Extract of Example <1-2> 10.0 1,3-butylene glycol 7.0 glycerin 1.0 D-panthenol 0.1 Plant extract 3.2 Magnesium aluminum silicate 0.3 PEG-40 Stearate 1.2 Stearic acid 2.0 Polysorbate 60 1.5 Chin type glyceryl stearate 2.0 Sorbitan sesquioleate 1.5 Cetearyl Alcohol 3.0 Mineral oil 4.0 Squalane 3.8 Carlylic / Capric Triglycerides 2.8 vegetable oil 1.8 Dimethicone 0.4 Dipotassium glycylizate a very small amount Allantoin a very small amount Sodium hyaruronate a very small amount Tocopheryl acetate Quantity Triethanolamine Quantity Preservative Quantity Flavoring agent Quantity Purified water Balance

< Manufacturing example  2> Sentipedgrass  Extract, its Fraction  Or above From fractions  Preparation of medicament for skin wrinkles containing separated active fraction as an active ingredient

<2-1> Preparation of Syrup

The syrup containing the centipede glass ethyl acetate-20% methanol active fraction of <Example 2> as an active ingredient was prepared as shown in the following [Table 6].

Ingredient Content (parts by weight) Ethyl Acetate-20% Methanol Fraction of <Example 2> 2 saccharin 0.8 Party 25.4 glycerin 8 Spices 0.04 ethanol 4 Sorbic acid 0.4 Distilled water 60

<2-2> Preparation of Tablet

The tablet containing the centipede glass ethyl acetate-20% methanol active fraction of <Example 2> as an active ingredient was prepared as shown in the following [Table 7].

Ingredient Content (parts by weight) Ethyl Acetate-20% Methanol Fraction of <Example 2> 250 Lactose 175.9 Potato starch 180 Colonde silicic acid 32 10% gelatin solution 5 Potato starch 160 talc 50 Magnesium stearate 5

250 parts by weight of the ethyl acetate-20% methanol active fraction of <Example 2>, 175.9 parts by weight of lactose, 180 parts by weight of potato starch, and 32 parts by weight of colloidal silicic acid were mixed. 10% gelatin solution was added to the mixture, which was then ground and passed through a 14 mesh sieve. This was dried and the mixture obtained by adding 160 parts by weight of potato starch, 50 parts by weight of talc and 5 parts by weight of magnesium stearate was prepared as a tablet.

< Manufacturing example  3> Sentipedgrass  Extract, its Fraction  Or above From fractions  Preparation of health food for skin wrinkle improvement containing separated active fraction as active ingredient

Foods containing the centipede glass water extract of Example <1-3> as an active ingredient were prepared as follows.

<3-1> Production of flour food

0.5 to 5.0 parts by weight of the extract of Example <1-3> was added to the flour, and bread, cake, cookies, crackers, and noodles were prepared using the mixture.

<3-2> soup  And juicy ( gravies Manufacturing

0.1 to 5.0 parts by weight of the extract of Example <1-3> was added to soups and broths to prepare meat products for health promotion, soups and noodles for noodles.

<3-3> Ground Beef  Produce

10 parts by weight of the extract of Example <1-3> was added to the ground beef to prepare a ground beef for health promotion.

<3-4> Dairy products ( dairy products Manufacturing

5-10 parts by weight of the extract of Example <1-3> was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<3-5> Solar  Produce

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh.

Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer.

The extract of Example <1-3> was concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by drying with a spray and a hot air dryer was pulverized with a particle size of 60 mesh to obtain a dry powder.

The grains, seeds, and extracts of Example <1-3> prepared above were prepared by combining the following ratios.

Cereals (30 parts by weight brown rice, 15 parts by weight brittle, 20 parts by weight of barley),

Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds)

Extract (3 parts by weight) of Example <1-3>,

(0.5 part by weight),

Foxglove (0.5 part by weight)

< Manufacturing example  4> Sentipedgrass  Extract, its Fraction  Or above From fractions  Preparation of a beverage for skin wrinkles containing the separated active fraction as an active ingredient

<4-1> Health drink  Produce

Homogeneous components such as liquid fructose (0.5%), oligosaccharides (2%), sugar (2%), salt (0.5%), water (75%) and 5 g of the extract of Example 1-3 of the present invention are homogeneous. After sterilization and instant sterilization, it was prepared by packaging in small packaging containers such as glass bottles and plastic bottles.

<4-2> Production of vegetable juice

5 g of the extract of Example <1-3> was added to 1,000 ml of tomato or carrot juice to prepare vegetable juice.

<4-3> Preparation of fruit juice

1 g of the extract of Example <1-3> was added to 1,000 ml of apple or grape juice to prepare a fruit juice.

As described above, the centifed glass extract of the present invention, its fractions or the active fractions separated from the fractions have excellent elastase inhibitory effect and have an excellent effect on wrinkle improvement, so that cosmetics, external preparations for skin wrinkles, drugs or health It can be usefully used for food development and product production.

1 is a diagram illustrating a process for preparing an extract of centipedegrass, fractions thereof, and active fractions separated from the fractions.

2 is a diagram showing high performance liquid chromatography identifying secondary metabolites present in centipedegrass methanol extract.

Figure 3 is a diagram showing high performance liquid chromatography identifying secondary metabolites present in centifed glass ethyl acetate-20% methanol active fractions.

FIG. 4 is a diagram showing high performance liquid chromatography identifying secondary metabolites present in centifed glass ethyl acetate-50% methanol active fraction.

FIG. 5 is a diagram showing high performance liquid chromatography identifying secondary metabolites present in centifed glass ethyl acetate-70% methanol active fraction.

Figure 6 is a graph showing the inhibition rate of elastase according to the concentration of centipede glass methanol extract or corn beard methanol extract.

7 is a graph showing the elastase inhibition rate of the active fraction (ethylacetate-20% methanol, -50% methanol and -70% methanol) separated from the centipedes glass methanol extract and fractions thereof.

Claims (8)

delete delete delete Skin containing the ethyl acetate fraction obtained by dissolving the extract of centifedgrass with water, lower alcohols of C ₁ to C ₄ or a mixed solvent thereof in water and then systematically fractionating it in the order of n-hexane and ethyl acetate. Wrinkle improvement cosmetic composition. delete The ethyl acetate fraction obtained by dissolving the extract of centifedgrass with water, lower alcohols of C ₁ to C ₄ or a mixed solvent thereof in water and then systematically fractionating it in the order of n-hexane and ethyl acetate was further adsorbed on column chromatography. And 20%, 50% or 70% methanol as an eluting solvent, and containing the active fraction separated as an active ingredient. delete delete

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