KR101070886B1 - A composition comprising pachydictyon coriaceum extract - Google Patents

Abstract

The present invention relates to a composition comprising the extract Pachydictyon coriaceum . The participating moth can be usefully applied to cosmetics, medicines and foods for this purpose by showing a whitening effect, wrinkle improvement effect, hair growth promoting effect, anti-inflammatory effect, antioxidant effect, anti-obesity effect and the like.

Participation, wrinkle, hair growth, anti-inflammatory, anti-obesity

Description

A composition containing the extract of bamboo flakes {A COMPOSITION COMPRISING PACHYDICTYON CORIACEUM EXTRACT}

The present invention relates to a composition comprising the bamboo shoots extract. Particularly, the bamboo bark extract of the present invention is excellent in whitening effect, wrinkle improvement effect, immunomodulatory effect, antioxidant effect, anti-obesity effect and hair growth effect, the present invention is a cosmetic composition comprising the bamboo bark extract extract , Pharmaceutical compositions and food compositions.

Human skin color is determined by the concentration and distribution of melanin inside the skin. Melanin pigment produced by melanocytes of human skin is a phenolic polymer having a complex form of black pigment and protein, and plays an important role in blocking skin damage caused by ultraviolet rays. It is reported that the action of tyrosinase in melanocytes is the most important for melanin biosynthesis, and tyrosinase is a type of amino acid tyrosine (DOPA) which is an intermediate of melanin polymer production. It also plays a key role in skin blackening by converting it to dopaquinone.

In general, skin wrinkles are known to be produced by a combination of several factors, such as the skin's water content, collagen content, and the ability to act on the environment. Among these factors, the most important influence on the formation of wrinkles is the expression and activity of collagenase, a collagen degrading enzyme that reduces collagen production and collagen content.

Hormonal imbalance is aggravating due to environmental pollution and automobile exhaust gas. As a result, the incidence of hair loss increases, the age of incidence decreases, and incompatibility of the skin immune system is caused, resulting in various skin diseases. In addition, the population suffering from obesity is rapidly increasing despite the age of children due to the changes in the diet of meat and meat.

Therefore, the development of materials that can effectively solve various phenomena, such as blemishes or freckles, wrinkles caused by ultraviolet rays, obesity, immune imbalance, hair loss, etc., which are not only aesthetical but also serious social problems, have been studied in depth. The present inventors have been working to solve the above problems, while confirming the efficacy of the participating bamboo nettle extract to complete the present invention.

One object of the present invention is to provide a composition for improving skin conditions such as skin inflammation, wrinkles, aging of the skin, and hair growth, comprising the extract of Participatum.

It is another object of the present invention to provide a pharmaceutical composition for hair growth, anti-obesity and anti-inflammatory, which comprises the extract of horseradish spp.

Another object of the present invention is to provide an anti-obesity food composition comprising the extract of the bamboo shoots.

In one aspect, the present invention relates to a composition comprising the extract of bamboo flakes.

Participation of the present invention ( Pachydictyon coriaceum ) refers to the brown algae belonging to the Phaeophyta (Phaeophyceae) Diktyotaaceae (Dictyotales) Dicttyotaceae. The participant's body spreads somewhat fan-shaped and grows up to 30-40 cm in height, and branches into a seven-phase image. In addition, the lower part forms a short neck and has a brown hair. The branch is the object, the width is 7 to 15 mm, and the edge is the leading edge. The role of skin composition such as whitening and anti-wrinkle effect of this marine plant is not known with regard to the true leather net horse which has these characteristics.

Participating ginseng extract of the present invention can be prepared according to a conventional method for preparing seaweed extract. Specifically, it may be prepared by a cold extraction method, a hot extraction method or a heat extraction method, it may be prepared using a conventional extraction device, ultrasonic grinding extractor or fractionator.

In one specific embodiment, the dried or undried bamboo grits are pulverized, immersed in the pulverized product by adding a solvent in a weight ratio of 10 to 100 times, and then filtered or centrifuged to obtain a supernatant liquid. Leather net foot extract can be prepared.

As another specific embodiment, water or an organic solvent is added to the participating bamboo grass, and at least 70 hours, preferably at least 80 degrees Celsius, more preferably at least 90 degrees Celsius, at least 1 hour, preferably at least 2 hours. More preferably 4 hours or more and the heated extract can be produced by filtration.

In preparing the extract, the extraction solvent used is not limited thereto, but may be water, an organic solvent or a mixed solvent thereof. The organic solvent is, for example, alcohols such as methanol, ethanol, isoprotanol, butanol, ethylene, acetone, ether, hexane, chloroform, ethyl acetate, DMF (N, N-dimethylformamide), DMSO (dimethylsulfoxide ) And mixtures thereof. Preferably the extraction solvent may be water or ethanol.

The extract extracted with the solvent is, but is not limited thereto, and may be further fractionated with a solvent selected from the group consisting of hexane, methylene chloride, acetone, ethyl acetate, ethyl ether, chloroform, water and mixtures thereof. The fractional solvent may be preferably at least one selected from the group consisting of aqueous ethanol, water, ethyl acetate and chloroform.

The prepared extract or the fraction obtained by performing the fractionation process can be filtered or concentrated or dried to remove the solvent, it can be carried out both filtration, concentration and drying. Specifically, the filtration may be performed using a filter paper or a reduced pressure filter, the concentration may be concentrated under reduced pressure using a reduced pressure concentrator, for example, a rotary evaporator, and the drying may be performed by, for example, a lyophilization method.

Therefore, the participated porridge root extract used in the present invention includes the prepared extract or fraction, and the extract or fraction may be one which completely removes moisture by concentrating and lyophilizing and completely removing the moisture. Brown algae extract or fractions may be used in powder form, or the powder may be used after being dissolved in distilled water or a common solvent.

Participating ginseng extract of the present invention exhibits an excellent whitening effect of inhibiting melanin production by inhibiting the activity of intracellular tyrosinase, has high stability, almost no change in the component content over time, and skin irritation. There are few side effects such as induction.

In addition, the participated bark nettle extract of the present invention exhibits an excellent wrinkle improvement effect through molecular mechanisms such as inhibiting collagenase activity and promoting collagen synthesis.

In addition, the porridge extract of the present invention is excellent in hair growth promoting effect. As a result of comparison with a commercially available hair regrowth in a specific implementation, it was found that the hair growth effect of the participating bamboo bark horse extract of the present invention is excellent. For convenience, the term "promoting hair growth" is used interchangeably with "preventing hair loss" or "promoting hair growth".

In addition, the porridge extract of the present invention is excellent in the antioxidant effect due to the excellent free radical removal effect. The correlation between many disease states and free radical damage is well established and many cellular components, including enzymes, ion channels, structural proteins and membrane lipids, are potential targets for reactive free radical species (Rice-Evans C). , Mol Aspects of Med 13 (1): 1-111 (1992). The reaction of these potential targets with free radicals impairs a range of cellular functions leading to pathological changes that ultimately kill cells. In addition, various diseases caused by physiological oxidation are disclosed in US Pat. Nos. 5,750,351, 5,773,209, 5,773,231, and 5,846,959. Examples of the disease include atherosclerosis, coronary artery disease, coronary artery restenosis, reperfusion injury, neurodegenerative diseases such as Parkinson's disease or Alzheimer's disease, stroke, cancer, aging, cardiovascular disease, osteoporosis, central nervous system disorders, peripheral vascular disease And difficulty breathing. Therefore, the bamboo bark extract of the present invention has an excellent antioxidant effect and can be usefully used for the prevention and treatment of diseases as described above.

In addition, the bamboo bark extract of the present invention has an anti-inflammatory effect because it has the effect of inhibiting COX activity.

In addition, the porridge extract of the present invention has the effect of inhibiting obesity.

As one specific embodiment, the present invention is a cosmetic composition for improving the skin condition comprising the participating moth extract, wherein the skin condition is any one or more of skin inflammation, wrinkles, aging and regrowth of skin cosmetics It relates to a composition.

In the present invention, "skin condition" means skin inflammation, wrinkles, aging of the skin and hair growth.

Participants of the present invention is included in an amount of 0.01 to 20% by weight, preferably 0.01 to 10% by weight, more preferably 0.01 to 5% by weight relative to the total weight of the composition.

The components included in the cosmetic composition of the present invention include components conventionally used in cosmetic compositions in addition to the participating bamboo grass extract as an active ingredient. Conventional adjuvants such as, for example, antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings, and carriers. In addition, the cosmetic composition may further include a skin absorption promoting substance to enhance the effect.

The cosmetic composition of the present invention may be prepared in any formulation commonly prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing , Oils, powder foundations, emulsion foundations, wax foundations and sprays, and the like, but are not limited thereto. More specifically, it can be manufactured in the form of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

When the formulation of the present invention is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components. Can be.

In the case where the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / Propane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Soluble cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

In another specific embodiment, the present invention relates to a pharmaceutical composition for hair growth, anti-obesity and anti-inflammatory, comprising the extract of Participating Bacteria.

Participants of the present invention is included in an amount of 0.01 to 20% by weight, preferably 0.01 to 10% by weight, more preferably 0.01 to 5% by weight relative to the total weight of the pharmaceutical composition.

The pharmaceutical composition of the present invention may further contain at least one active ingredient exhibiting hair growth promoting, obesity inhibiting and / or anti-inflammatory effect in addition to the participating bamboo grass extract.

In addition, the pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier conventionally used in the preparation of the pharmaceutical composition. Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. The pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives and the like in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, humans by various routes such as oral or parenteral, such as oral, rectal or intravenous, muscle, subcutaneous, intrauterine dura or cerebrovascular vessels. It can be administered by internal injection. It is preferably applied in the form of topical application during parenteral administration, more preferably by application.

Suitable dosages of the pharmaceutical compositions of the present invention may vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be. The dosage of the pharmaceutical composition of the present invention may be administered once or several times a day in an oral dosage form of 0.1-100 mg / kg on an adult basis. It is recommended to apply 1 to 5 times a day in an amount of 3.0 ml to continue for 1 month or more. However, the dosage does not limit the scope of the present invention.

The pharmaceutical compositions of the present invention may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporation into a multi-dose container. The formulation may be used in any form suitable for pharmaceutical preparations, including powders, granules, tablets, capsules, suspensions, emulsions, syrups, oral formulations such as aerosols, external preparations such as ointments, creams, suppositories, and sterile injectable solutions. It may further comprise a dispersant or stabilizer.

In addition, the pharmaceutical compositions of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormonal therapy, chemotherapy and biological response modifiers for promoting hair growth, inhibiting obesity and anti-inflammatory.

As another specific aspect, as another aspect, this invention relates to the food composition for anti-obesity which contains the extract of horseradish root.

Participants of the present invention is included in an amount of 0.01 to 20% by weight, preferably 0.01 to 10% by weight, more preferably 0.01 to 5% by weight relative to the total weight of the food composition.

Participants of the present invention may be added to health food for the purpose of inhibiting obesity. When using the ginseng extract of the present invention as a food additive, it can be added as it is, or used together with other food or food ingredients, can be appropriately used in accordance with conventional methods. The mixed amount of the active ingredient may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment). Generally, in the preparation of food or beverages, the composition of the present invention is added in an amount of up to 15% by weight, preferably up to 10% by weight relative to the raw material. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of health control, it may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety.

There is no particular limitation on the kind of food. Examples of the food to which the substance may be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, tea, and drinks. Alcoholic beverages and vitamin complexes, and includes all of the health foods in the conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The natural carbohydrates described above may be used as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and natural sweeteners such as dextrin and cyclodextrin, and synthetic sweeteners such as saccharin and aspartame. . The proportion of the natural carbohydrate is generally about 0.01 to 0.4 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 composition of the present invention includes various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, And a carbonation agent used for the carbonated beverage. In addition, the composition of the present invention may contain a 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.

On the other hand, in the specific practice of the present invention, the skin accumulation stimulation test results for the participating bamboo grass extract extract was found to be a natural substance harmless to the human body. Therefore, the bamboo shoots extract of the present invention has little toxicity and side effects, so that it can be used safely even when used for a long time, and in particular, it can be safely applied to cosmetics, drugs and food compositions as described above.

Participating bamboo extract of the present invention has excellent whitening effect, inhibits collagenase activity and promotes collagen synthesis by inhibiting melanin production by inhibiting the activity of intracellular tyrosinase compared to arbutin, which is used as a conventional skin whitening agent. Through molecular mechanisms such as excellent wrinkle improvement, anti-inflammatory effect, hair growth promoting effect, antioxidant effect and anti-obesity effect. In addition, the porridge extract of the present invention can be safely applied to cosmetics, drugs and foods because there is no cytotoxicity and skin side effects.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, and the scope of the present invention is not limited by these examples in accordance with the gist of the present invention to those skilled in the art. Will be self-evident.

Example 1 Preparation of Participation Grain Extract

1-1. Preparation of Water Soluble Extract

1 kg of fully dried bamboo sprouts was placed in 3 l of distilled water, and extracted in a extractor equipped with a cooling condenser at 15 to 37 ° C. for 14 days. The extract was filtered through a 400 mesh filter cloth, left to mature at 5 to 15 ° C. for 7 days, and then filtered using Whatman No. 2 filter paper. These plant extracts were concentrated under reduced pressure in a distillation apparatus equipped with a cooling condenser to obtain 29 g, 30 g, and 29 g (dry weight), respectively.

1-2. Preparation of Ethanol Soluble Extract

1 kg of fully dried bamboo sprouts were placed in 3 l of ethanol and extracted for 14 days at 15 to 37 ° C. in an extractor equipped with a cooling condenser. The extract was filtered through a 400 mesh filter cloth, left to mature at 5 to 15 ° C. for 7 days, and then filtered using Whatman No. 2 filter paper. These plant extracts were concentrated under reduced pressure in a distillation apparatus equipped with a cooling condenser to obtain 31 g, 39 g, and 37 g (dry weight), respectively.

1-3. Preparation of Methanol Soluble Extract

1 kg of fully dried bamboo flakes were placed in 3 l of methanol and extracted for 14 days at 15 to 37 ° C. in an extractor equipped with a cooling condenser. The extract was filtered through a 400 mesh filter cloth, left to mature at 5 to 15 ° C. for 7 days, and then filtered using Whatman No. 2 filter paper. These plant extracts were concentrated under reduced pressure in a distillation apparatus equipped with a cooling condenser to obtain 38 g, 41 g, and 39 g (dry weight), respectively.

Example 2 measurement of melanin production inhibitory effect

B16 melanoma cells (Korea Cell Line Bank) were used to measure the inhibitory effect of melanin production by the extracts of the bark horseweed and compared with the inhibitory effect of melanin production by arbutin known to inhibit melanin production. It was.

In this experiment, murine melanoma (B-16 F10) cells (Korea Cell Line Bank) were used in DMEM medium containing 10% fetal bovine serum (FBS) in a 6-well plate at 1 × 10 ⁵ per well. After inoculation with dogs, the cells were incubated at 5% CO ₂ and 37 ° C until at least about 80% were attached to the bottom of the well. After incubation, the medium was removed, and 100 parts of arbutin (Arigin, Sigma) and the participant extract of Example 1-2 were replaced with DMEM medium diluted to 10 ppm, 100 ppm and 500 ppm of non-cytotoxicity. , 5% CO ₂ , was incubated for 3 days at 37 ℃. Thereafter, the medium was removed, and the collected cells were washed with PBS (phosphate buffer saline), which was treated with trypsin to recover the cells. The recovered cells were measured by using a hematocytometer (Tiefe Depth Profondeur 0.100 mm, Paul Marienfeld GmbH & Co. KG, Germany) to measure the cell number, centrifuged at 5,000 to 10,000 rpm for 10 minutes, and then the supernatant was removed. It was removed to obtain pellets. The cell pellet was dried at 60 ° C., and then 100 μl of 1 M sodium hydroxide solution containing 10% DMSO was added to obtain intracellular melanin in a 60 ° C. thermostat. Using this solution, the absorbance was measured at 490 nm using a microplate reader (Bio-Tek ELx8081U, USA) to determine the amount of melanin per cell. The results are shown in Table 1 below.

sample Treatment Concentration (ppm) Melanin production inhibition rate (%) Arbutin 100 20 Participated Bamboo Net Extract 10 7 Participated Bamboo Net Extract 100 30 Participated Bamboo Net Extract 500 58

As shown in Table 1, it was found that the participant extract was higher in melanin production than arbutin, and the higher the treatment concentration of the participant extract was higher in melanin production.

Example 3 Measurement of Inhibitory Effects of Intracellular Tyrosinase Activity by Participated Extracts

Using melanoma cells (B16 melanoma cells) to measure the inhibitory effect of melanin production by the extracts of the bark horseweed, and compared with the inhibitory effect of intracellular tyrosinase activity by arbutin known to inhibit melanin production It was.

This experiment was performed by inoculating murine melanoma (B-16 F1) cells in 6-well plates at 1 × 10 ⁵ per well in DMEM medium containing 10% FBS (fetal bovine serum). The cells were incubated at% CO ₂ , 37 ° C. until the cells adhered at least about 80% to the bottom of the well. After incubation, the medium was removed, and 100 parts of arbutin (Arigin, Sigma) and the participant extract of Example 1-2 were replaced with DMEM medium diluted to 10 ppm, 100 ppm and 500 ppm of non-cytotoxicity. , 5% CO ₂ , was incubated for 3 days at 37 ℃. Thereafter, the medium was removed, and the collected cells were washed with PBS (phosphate buffer saline), which was treated with trypsin to recover the cells. The recovered cells were measured by using a hematocytometer, and then centrifuged at 5,000 to 10,000 rpm for 10 minutes, and then the supernatant was removed to obtain pellets. The cell pellet was pulverized using lysis buffer, and then centrifuged at 12,000 rpm for 10 minutes to collect supernatant. Using this solution, absorbance at 492 nm was measured with a microplate reader to determine cell-specific allowance tyrosinase activity. The results are shown in Table 2 below.

sample Treatment Concentration (ppm) Inhibition rate of intracellular tyrosinase activity (%) Arbutin 100 15 Participated Bamboo Net Extract 10 6 Participated Bamboo Net Extract 100 27 Participated Bamboo Net Extract 500 52

As shown in Table 2, it was found that the participant extract was higher in intracellular tyrosinase inhibition rate than arbutin. In addition, it was found that the degree of inhibition of tyrosinase activity of the participating bamboo bark horse extract was concentration-dependent.

Example 4 Evaluation of Whitening Effect at Animal Level

Similar to humans, the whitening effect of participating moth extract was measured using brown malmoto (Tortoiseshell guinea pigs; Brown guinea pigs, Charles River Laboratories, Inc.), which is known to cause pigmentation by ultraviolet rays.

In order to cause pigmentation due to ultraviolet (UV) in the brown molemoto, a light shielding aluminum foil having a square window of 3 × 3 cm ² is adhered to the skin from which the hair of the brown molemoto coordination is removed, and then SE lamp ( Ultraviolet rays were irradiated with a wavelength of 290-320 nm (Toshiba) (total amount of irradiation energy = 1350 mJ / cm ² ). After the UV irradiation, the aluminum foil was peeled off, and the samples (seedle bran extract and arbutin) were applied in the following manner. Pigmentation appeared 2-3 days after UV irradiation and reached a peak after about 2 weeks, from which each sample was applied.

The number of coatings was continued once or twice a day for 50 days. The sample was dissolved and diluted using a specific solvent (propylene solvent: ethanol: water = 5: 3: 2: mixed solvent), and applied with a cotton swab, and prepared a control site for necessarily applying the solvent to other sites. Along with the determination of effects, the cumulative stimulus was also observed.

The effect was determined by measuring the degree of black and white of the skin using a color difference meter (CR2002 chromameter, Minolta, Japan), the results are shown in Table 3 below. L ^* A ^* B ^* color system is used to display color, and L ^* value was used as an index in this invention. L ^* value was calibrated with a standard whiteboard, and L ^* value was measured 5 times or more at one site, and pigmentation was equalized. The difference (ΔL ^* ) of the skin color at the start and completion of the application was determined (see Equation 1 below), and the effect was determined using this value. The results are shown in Table 3 below.

△ L ^* = L ^* value after 00 days application - L ^* value of the coating start date

Comparing the ΔL ^* value with respect to the sample application site and the control site, the effect of the whitening material can be seen.

sample Treatment Concentration (%) Whitening effect (△ L) Participated Bamboo Net Extract 0.2 0.38 Participated Bamboo Net Extract 1.0 0.62 Arbutin 1.0 0.54 Control group - 0.35

As shown in Table 3 above, the participating bamboo grass extract showed a concentration-dependent whitening effect. In addition, Participants extract showed better whitening effect than Arbutin, and cumulative irritation was not observed during application test.

Example 5 Safety Confirmation Test of Human Participate

5-1: Preparation of External Skin Agents Containing Participant Extract

In order to confirm that the participating bamboo bark extract was found to be excellent in the whitening effect as described above, it is also safe for human skin, a skin external preparation containing the participating bark extract and arbutin was prepared and skin safety verification experiment was conducted. It was.

The skin external preparations containing the extracts of bark horseweed and arbutin were prepared according to the ingredients in Table 4 below. In the following Table 4, the control group is a skin external preparation that does not include a tyrosinase inhibitor, and test group 1 and test group 2 are external skin preparations including the extracts of moths and arbutin.

For the preparation of external skin preparations, purified water, serine and butylene glycol were mixed and dissolved at 70 ° C. (aqueous part), and the other components except the three components and trimethanolamine were dissolved at 70 ° C. (oil phase part). The oil part was added to the water part and stirred with a homomixer (Tokushu Kika, Japan) to emulsify first, to which trimethanolamine was finally added. After removing the bubbles generated in the mixed solution, and cooled to room temperature to prepare a skin external preparation.

ingredient content (%) Control group Test group 1 Test group 2 Purified water 72 72 72 glycerin 8.0 8.0 8.0 Butylene glycol 4.0 4.0 4.0 Participated Bamboo Net Extract 0 1.0 0 Arbutin 0 0 1.0 Caprylic Capri Triglyceride 8.0 8.0 8.0 Squalane 5.0 5.0 5.0 Cetearyl Glucoside 1.5 1.5 1.5 Sorbitan stearate 0.4 0.4 0.4 Cetearyl Alcohol 1.0 1.0 1.0 Trimethanolamine 0.1 0.1 0.1 Gross weight 100 100 100

5-2: Accumulated skin irritation test

Participating bamboo root extract was irritated to the skin by subjecting the upper forearm to a total of nine 24-hour cumulative patches every other day for 30 healthy adults using the skin preparations prepared in Example 4-1. It was measured whether or not to give.

The patch method used the fin chamber (Finn chamber, Epitest Ltd, Finland). 15 µl of each of the external skin preparations was added dropwise to the chamber, followed by patching. The degree of response to the skin each time was scored using Equation 2 below, and the results are shown in Table 5 below.

Average responsiveness = [{(response x responsiveness) / (total number of subjects x highest score (4 points))} x 100] ÷ number of tests (9 times)

At this time, ± 1 point, + 2 points, + + 4 points in the reaction degree, and when the average reactivity is less than 3 was determined as a safe composition.

In Test Tables 1 and 2 in Table 5, the number of people corresponding to ±, +, and ++ was 3, 0, and 1, respectively, and the rest did not appear. When calculated according to the formula described above, the average reactivity was 0.37, which was 3 or less, which was determined to be a safe composition.

As shown in Table 5, the external skin preparations containing the extracts of the Participating bark horseweed extract (Test Group 1) did not show a distinct cumulative stimulation pattern as the skin external preparations including the control group or arbutin and were determined to be safe for human skin.

Example 6 Antioxidant Effect of Participating Grain Extract

Superoxide dismutase (SOD) is known as an antioxidant that converts superoxide anions into H ₂ O ₂ and O ₂ . This experiment is a method of evaluating the antioxidant capacity of a sample by observing the rate at which the superoxide anion generated by Xanthine oxidase was removed by the sample used in the test. The experiment was carried out using a kit purchased from Dojindo (Japan) (SOD Assay Kit-WST), and was tested according to the manufacturer's protocol. In brief, the effect was determined by observing the removal rate of superoxide anion by the sample, the presence or absence of antioxidant function. The experimental results are shown in Table 6.

sample Treatment Concentration (ppm) Superoxide Anion Removal Rate (%) Participated Bamboo Net Extract 5 7 Participated Bamboo Net Extract 10 20 Participated Bamboo Net Extract 100 38

As shown in Table 6, it was found that the participating bamboo bark extract of the present invention has an antioxidant effect in a concentration-dependent manner.

Example 7 Evaluation of Anti-Inflammatory Effect of Participation

In order to examine the anti-inflammatory effects of the extracts of the fungi, we conducted COX activity inhibitory experiments according to the conventional method in THP-1 cells, which are human monocytes. COX activity was measured according to Method in Enzymology 43: 9 (1994) published by FJ Van de Ouderaa and M Buytenhek . THP-1 cell line (Korea Cell Line Bank) was cultured and the cells were dispensed in 24-well plates. The incubation volume per well was adjusted to 500 μl, and 2 μl of sample dissolved in LPS (lipopolysaccharide, Sigma) (1 μg / ml) and the solvent shown in the following table was added, and then incubated under the same conditions for 24 to 48 hours. After 24 to 48 hours, 1 μl of calcium ionophore (Sigma) and [1-14C] arachidonic acid (in EtOH, 0.1 μCi / ml, Sigma) were added to each well and incubated for 10 minutes under the same conditions. After incubation, citric acid was added to each well, and the pH was adjusted to 3.5. Then, 500 μl of each culture solution was aliquoted into a microcentrifuge tube, and 700 μl of ethyl acetate was added thereto, followed by shaking extraction for 10 minutes. 500 µl of ethyl acetate layer was taken and concentrated for 20 minutes using a speed vacuum dryer. The concentrated residue was dissolved in 20 µl of ethyl acetate and applied to an TLC plate with an authentic standard. Radioactivity bands were identified by authentic eicosanoid standards, and the radioactivity of the identified bands was measured using a BAS 2000 bio-imaging analyzer (Fuji, Japan) (Table 7). .

sample COX activity (%) LPS (1 μg / ml) 100 LPS (1 ㎍ / ml) + Participated Horseweed Extract (5 ppm) 92 LPS (1 ㎍ / ml) + Participated Horseweed Extract (10 ppm) 85 LPS (1 ㎍ / ml) + Participated Horseweed Extract (100 ppm) 63

As shown in Table 7, it can be seen that the participating bamboo bark extract of the present invention effectively inhibits the activity of COX as the treatment concentration is increased, and it can be seen that there is an effect of inhibiting inflammation.

Example 8: Evaluation of immunosuppressive ability of the extract

Whether or not the extract of the bark of the present invention inhibits an immune related response related to inflammation was investigated through an interleukin-2 luciferase reporter activity test. The interleukin-2 promoter is reported to play an important role in the production of immune-related cytokines associated with inflammation. The following method was used to measure interleukin-2 luciferase activity. After dispensing 1 × 10 ⁶ cells into 6 wells of Jurkat cells (Korea Cell Line Bank), which is a human T lymphocyte cell line, interleukin-2 lucifera using a superperfect transfection reagent (In vitrogen) was used. Aze reporter plasmid DNA (IL-2 luciferase reporter plasmid DNA) was transfected. After 24 hours after transfection, PHA (Phytohemaglutinin, Sigma) (100 ng / ml) was treated to activate Jurkat cells and each sample was treated by concentration. After 24 hours, cells were collected and luciferase activity was measured using a luminometer (Berthold Technologies GmbH & Co.KG, Germany). The results are as follows.

sample Interleukin-2 Luciferase Activity PHA (100 ng / ml) 100 PHA (100 ng / ml) + Participated Horseweed Extract (5 ppm) 87 PHA (100 ng / ml) + Participated Horseweed Extract (10 ppm) 76 PHA (100 ng / ml) + Participated Horseweed Extract (100 ppm) 50

As shown in Table 8, it was found that the participating bamboo grass extract has an immunomodulatory function by inhibiting the expression of interleukin-2 as the treatment concentration increases.

Example 9 Measurement of Wrinkle Improvement Effect of Participants

The anti-wrinkle effect can usually be measured as collagen biosynthesis and collagenase degradation inhibitory effect and clinical trial in humans.

Human normal fibroblasts (Pacific) were inoculated into 6-well microplates containing DMEM medium (2 × 10 ⁵ cells / well) in a 2% CO ₂ incubator at 37 ° C. Time incubation. After 24 hours, the medium was removed from each well, samples were treated by concentration and then incubated again for 24 hours. After 24 hours, cell media were collected to prepare samples.

Collagen synthesis amount was measured by using the collagen measurement kit (Procollagen Type I C-peptide EIA kit (MK101), Takara, Kyoto, Japan) according to the manufacturer's protocol. The amount of Type I C-peptide (PICP) was measured and analyzed.

An antibody against collagenase was used as a method for measuring the activity of collagenase, an enzyme that degrades collagen. Collagenase activity was measured using a type I collagenase assay kit (Amersham Biosciences, RPN2629) and absorbance was measured with an ELISA reader (Bio-Tek ELx808 ™ Series Ultra Microplate Reader, UK). The measured standard values were expressed in the form of mean ± standard deviation and tested for significance by t-test using the SPSS / PC + program. The results are shown in Table 9 below.

Test Items Participation
Extract (1 ppm) Participation
Extract (5 ppm) Participation
Extract (10 ppm) Collagen Synthesis Growth Rate 5 17 29 Collagenase inhibition rate 7 15 26

As shown in Table 9, the Participants extract increased collagen synthesis concentration-dependently, and also inhibited collagenase activity concentration-dependently. Therefore, it was found that the participating bamboo grass extract has an effect of improving wrinkles.

Example 10: Anti-obesity Effect of Participating Grain Horsetail Extract (Oral)

10-1: Liposomal Formulation Preparation

One). Lecithin, ethanol, ceramide, cholesterol ester, squalane, caprylic capric triglyceride and phytosterol in the oil phase components were measured and the temperature was raised (60-65 ° C.), followed by dissolving the azi mixer at 150 rpm.

2). The moisturizing ingredients glycerin, hyaluronic acid, and sorbitol are measured in the aqueous phase to raise the temperature (60 to 65 ° C.), and then the azi mixer is stirred at 150 rpm to dissolve.

3). Just before emulsification, add the active ingredient bamboo mat powder (0.1%, 1%).

4). The temperature of the water phase component and oil phase component is maintained at 60 to 65 ° C., while the azi mixer of the water phase component is stirred at 100 rpm and the homo mixer at 3,000 rpm, and the oil phase component is slowly added (emulsion time: 5 min).

5). After emulsification, the contents are passed through a cooler and cooled to 30 ℃, and then passed through a microfluidizer three times.

6). After stirring, the thickener is gradually added to prevent tangling, and the mixture is cooled to 40 ° C., and then preservatives and additives (tocopherol, etc.) are added to disperse and dissolve the azi mixer at a speed of 150 rpm.

7). A vacuum pump is operated to degas the mixing tank for 10 minutes in a continuous, bleeding manner, by vacuuming the inside of the mixing tank.

8). After cooling to 28 ~ 30 ℃, the semi-finished product is passed through a 200 mesh filter screen to remove foreign substances, and then stored in a storage container at room temperature (25 ± 2 ℃).

10-2: Anti-obesity effect by oral administration

The obesity suppression test was carried out with 7 weeks old ICR male mice (Chicks River Co., Ltd., Japan) for 1 week, and then used in experiments in 7 groups. Animals were raised in a constant temperature room set at a temperature of 23 ± 1 ℃, a humidity of 55 ± 5% and an illumination time of 12 hours / day. In addition, water was freely consumed. Participating ginseng extract was prepared as a liposome formulation (dissolved in 5% lecithin) using the preparation method of 10-1 to be 0.1%, 1%. Each sample solution was adjusted to a concentration of 0.1 ml per 10 g of mouse, and doses were 1.5 g / kg and 1 g / kg. In addition, the control group was administered in the same amount as a 5% lecithin emulsion. Mice were fasted from the day before administration and were forcibly administered once the next day. The test period was two weeks, and body weight and general symptoms were measured and observed.

solvent Days Weight (g) Untreated group Bamboo Flour Extract (0.1%) Participated Bamboo Net Extract (1%) Liposomes
Formulation 5 24 24 23 10 28 24 24 15 34 26 26

As can be seen in Table 10, the participating bamboo net extract showed an anti-obesity effect. It was observed that the higher the dosage concentration, the better the efficacy.

Example 11 Hair Loss Prevention and Hair Growth Effect of P.

In order to find out about the hair loss prevention and hair growth effect of the participating bamboo net horse according to the invention, the following experiment was performed.

1. Preparation of Hair Regrowth Composition

The hair restorer composition containing the bamboo flakes was prepared as the component content of the following table.

ingredient content(%) Participation 1.0 Preservative (Gramben) 0.8 Thickener (Xanthan-Gum) 0.3 Total weight 100

Using the external preparation for promoting hair growth, the hair loss site of 10 patients with alopecia due to weakening or radioactive exposure was applied twice a day for 3 months each for 3 months. As a control, moxidil sold in Hanmi Pharm was used. The results are shown in Table 12 below, and showed excellent effects such as the occurrence of new hair roots in eight hair loss patients.

Untreated group Moxidil Participated Bamboo Net Extract One 4 3

[Criteria]

4: high effect = newborn hair visible,

3: moderate effect = visible newborn hair (downy),

2: slightly effective = the number of hair loss decreases,

1: no effect = no change at all.

As can be seen in Table 12, when the participating bamboo bark horse extract of the present invention was applied, it showed a hair growth effect inducing the generation of new hair.

Participating ginseng extract of the present invention has skin inflammation suppression, wrinkle improvement, anti-aging, hair growth promotion, antioxidant and anti-obesity effect, and can be usefully contained in cosmetics, drugs and foods.

Claims (7)

A cosmetic composition for improving skin condition, comprising the extract of bamboo flakes as an active ingredient, wherein the skin condition is any one or more of wrinkles and hair growth of the skin. The group of claim 1 wherein the composition consists of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray A composition characterized by having a formulation selected from. A pharmaceutical composition for regrowth comprising the extract of horseradish spp. As an active ingredient. A pharmaceutical composition for anti-obesity, comprising asparticulate extract. delete Health functional foods for improving obesity, including asparticulate extract. The composition according to any one of claims 1 to 4 and 6, wherein the participating bamboo grass extract is included in an amount of 0.01 to 10% by weight based on the total weight of the composition.