JP2006083070A - Active oxygen scavenger, bleaching agent, anti-inflammatory agent, collagenase activity inhibitor or collagenase production inhibitor comprising calycinal extract of diospyros kaki l as active ingredient - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To develop a method for effectively utilizing a calycinal part of Diospyros kaki L in industry. <P>SOLUTION: An active oxygen scavenger, a bleaching agent, an anti-inflammatory agent, a collagenase activity inhibitor or a collagenase production inhibitor comprising a calycinal extract of the Diospyros kaki L as an active ingredient is provided. A method for effectively carrying out prophylaxis or amelioration of photodegradation caused by exposure to ultraviolet light comprising using the active oxygen scavenger, bleaching agent, anti-inflammatory agent, collagenase activity inhibitor or collagenase production inhibitor is provided. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to an active oxygen scavenger, a whitening agent, an anti-inflammatory agent, a collagenase activity inhibitor or a collagenase production inhibitor. The present invention also relates to a method for preventing or improving photoaging caused by ultraviolet exposure.

  Japanese persimmon is produced at about 290,000 tons in Japan, and about 20,000 tons are produced in Gifu, which is famous for the production of Fuyu persimmon. Processed foods using persimmon fruits and persimmon fruits are widely used as food. However, the present state is that most of the cocoon portion of the cocoon is discarded as waste in processed foods and the like, and it is simply treated as trash other than processed foods. Therefore, it is necessary to devise a new method for industrially utilizing the cocoon part of the waste, which is waste, from the viewpoint of green chemistry that effectively uses what is obtained from nature.

  The problem to be solved is the development of an effective use method of the heel part of the cocoon in the industry.

  As a result of examining the effective utilization method of the cocoon part of the cocoon, the present inventors have found that the cocoon extract has excellent active oxygen scavenging action, whitening action, anti-inflammatory action, collagenase activity inhibitory action and collagenase production inhibitory action. As a result, it was made possible to construct an active oxygen scavenger, a whitening agent, an anti-inflammatory agent, a collagenase activity inhibitor or a collagenase production inhibitor using the koji extract. As prior disclosure techniques related to acupuncture extract, it has been used as a traditional Chinese medicine to cure hiccups and vomiting, as well as phototoxicity inhibitors and cosmetics and foods containing the same (Patent Document 1), hair makeup. Although there are applications to hair and hair restoration agents (Patent Document 2), etc., the application to the active oxygen scavenger, whitening agent, anti-inflammatory agent, collagenase activity inhibitor or collagenase production inhibitor which is the present invention is novel. is there.

JP 2000-319154 A Japanese Patent Laid-Open No. 2002-20242

  On the other hand, exposure to ultraviolet rays contained in sunlight causes photoaging. Photoaging causes excessive degradation of the cell matrix and excessive melanin production, leading to marked pigmentation. Active oxygen generated by exposure to ultraviolet rays not only promotes DNA damage to cells, but also induces inflammatory cytokines to generate inflammation and increases the expression of collagenase that degrades collagen. Based on the above findings, the present inventors use an active oxygen scavenger, a whitening agent, an anti-inflammatory agent, a collagenase activity inhibitor or a collagenase production inhibitor containing the koji extract as an active ingredient, resulting in UV exposure. It has been found that it is possible to effectively prevent or improve photoaging.

  That is, the present invention provides an active oxygen scavenger, a whitening agent, an anti-inflammatory agent, a collagenase activity inhibitor, or a collagenase production inhibitor comprising a koji extract as an active ingredient. In addition, the present invention uses an active oxygen scavenger, a whitening agent, an anti-inflammatory agent, a collagenase activity inhibitor or a collagenase production inhibitor containing a koji extract as an active ingredient to effectively prevent or prevent photoaging due to UV exposure. It provides a way to improve.

  The active oxygen scavenger, whitening agent, anti-inflammatory agent, collagenase activity inhibitor or collagenase production inhibitor comprising the koji extract as an active ingredient of the present invention has an excellent active oxygen scavenging action, whitening action, anti-inflammatory action It has a collagenase activity inhibitory action and a collagenase production inhibitory action. For the purpose of adding the various actions described above, these agents can be used as additives for cosmetics or foods. And it is possible to prevent or improve photoaging resulting from exposure to ultraviolet rays by externally applying or taking the above-mentioned various drugs or cosmetics or foods containing these drugs.

  The koji extract used in the product of the present invention is obtained by appropriately pulverizing dried koji (Diospyros kaki L) koji in water, ethanol, propyl alcohol, isopropyl alcohol, butanol, isopropanol. Lower alcohol such as butanol, propylene glycol, 1,3-butylene glycol, 1,2-butylene glycol, 1,4-butylene glycol, 1,5-pentanediol, 1,2-pentanediol, 1,3-pentanediol , 1,4-pentanediol, 1,3,5-pentanetriol, glycerin, polyhydric alcohols such as polyethylene glycol (molecular weight 100 to 100,000), acetone, ethyl acetate, diethyl ether, dimethyl ether, ethyl methyl ether, dioxane, Low polarity solution such as acetonitrile, xylene, benzene, chloroform, carbon tetrachloride, phenol, toluene 1 type selected from acids (hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, citric acid, etc.), alkalis (sodium hydroxide, potassium hydroxide, calcium hydroxide, ammonia, etc.), etc. Or it is the extract obtained by extracting using 2 or more types of single solvents or mixed solvents. There is no restriction | limiting in particular in the varieties of the cocoon used, Any can be used if it belongs to Diospyros kaki on taxonomics.

As the extraction condition, the extraction temperature can be arbitrarily set as long as it is higher than the freezing point and lower than the boiling point of the solvent to be used. Specifically, it is in the range of −4 ° C. to 150 ° C. Preferably, the temperature range is 25 to 100 ° C., in which the components contained in the koji are not easily denatured or lost due to the reaction, and the extraction efficiency is relatively good. Further, the pressure applied for extraction is appropriately set within the range of 1.0 to 2.5 kg / cm ² . The weight ratio of the solvent to the raw material can be arbitrarily set within the range of 4: 1 to 1: 100, specifically, the raw material: solvent, but the weight ratio of 1: 5 to 1:20 is particularly preferred. preferable. The extraction time is not particularly limited except for an extremely short time such as within one hour, and is in the range of several days to several weeks.

  As the soot extract used in the present invention, it is also possible to use a further purified extract obtained from the above solvent and extraction conditions. Specific purification methods include filtration methods using filter paper, filter aids, membrane filters, etc., liquid separation operation methods, distillation methods, sublimation methods, precipitation methods, recrystallization methods, centrifugal separation methods, and vacuum concentration methods. , PH adjustment, salting out and the like, and one or more of these methods are selected.

  The active oxygen scavenger, whitening agent, anti-inflammatory agent, collagenase activity inhibitor or collagenase production inhibitor containing the persimmon extract of the present invention as an active ingredient, the persimmon extract itself as an active oxygen scavenger, whitening agent, In addition to being used as an inflammatory agent, collagenase activity inhibitor or collagenase production inhibitor, water, alcohols, polyhydric alcohols, surfactants, acids, alkalis, thickeners, excipients, preservatives, etc. What was added and adjusted to the extract can also be used as the drug. The shape of the various drugs of the present invention is usually liquid, and may be solid, powder, granule, pellet, gel, viscous liquid, and the like.

  In the present invention, an active oxygen scavenger, a whitening agent, an anti-inflammatory agent, a collagenase activity inhibitor or a collagenase production inhibitor containing the koji extract as an active ingredient can be used as an additive ingredient in cosmetics or foods. The cosmetic dosage form is arbitrary and is in the form of capsule, powder, granule, solid, liquid, gel, foam, emulsion, cream, ointment, sheet, mousse, powder dispersion, multilayer It forms a dosage form such as aerosol. Specifically, skin lotion, essence, whitening lotion, milk, whitening milk, cream, whitening cream, ointment, whitening ointment, lotion, whitening lotion, oil, pack and other basic cosmetics, soap, cleansing cream, cleansing Skin cleanser such as lotion, cleansing milk, face wash, shampoo, rinse, hair care cosmetics such as hair treatment, hair cream, hair spray, hair tonic, hair gel, hair lotion, hair oil, hair essence, hair water, hair wax Hair conditioner such as hair foam, hair growth agent, hair nourishing agent, hair loss prevention agent, one-part hair dye or two-part hair dye, hair color etc., permanent agent such as permanent wave agent and hair straightener Scalp or hair cosmetics such as hair and wave retention agents, foundations, white powder, funny, lipstick, blush, Shadow, eyeliner, mascara, eyebrow pencil, make-up cosmetics such as eyelashes, finishing cosmetics such as nail enamel, perfumes, toothpaste, mouthwash, mouthwash, and a bath agent, and the like. In addition, the amount of the drug to be added to the cosmetic is generally used in a concentration range of 0.0001% by mass or more in the total amount of the composition, preferably 0.01 to 50.0% by mass, particularly preferably 0.1 to 20%. It is in the range of mass%.

  In addition, the dosage form of food is also arbitrary, and dosage forms such as ampoules, capsules, powders, granules, pills, tablets, solids, liquids, gels, bubbles, emulsions, creams, etc. Make it. Specifically, health foods with the above dosage forms, oral compositions such as gums, candy and tablets, processed fishery products such as kamaboko and chikuwa, livestock products such as sausages and ham, Western confectionery, Japanese confectionery Noodles, raw noodles, Chinese noodles, boiled noodles, buckwheat noodles, sauces, soy sauce, sauce, sugar, honey, powdered candy, syrups such as syrups, spices such as curry powder, mustard powder, pepper powder, jam, marmalade, Chocolate spreads, pickles, pickled vegetables, sprinkles, processed vegetables and fruits such as canned and bottled vegetables, dairy products such as cheese, butter and yogurt, processed cereal products, fermented foods, miso soup, soup, fruits Juices, vegetable juices, whey drinks, soft drinks, alcoholic beverages and the like can be mentioned. In addition, the amount of the drug to be added to the food is generally used in a concentration range of 0.0001% by mass or more in the total amount of the composition, preferably 1.0 to 80.0% by mass, particularly preferably 5.0 to 50%. It is in the range of mass%.

  The method for preventing or improving photoaging due to ultraviolet exposure by externally or internally taking the cocoon extract as an active ingredient according to the present invention includes the above-mentioned drugs containing the cocoon extract as an active ingredient It is to apply cosmetics to be blended on the skin, scalp, etc. on a daily basis or to ingest a food to blend them on a daily basis. There is no restriction | limiting in particular about the amount of external application | coating or ingestion once, It is preferable to adjust suitably according to each user's physical condition, skin condition, environment, etc.

  The active oxygen scavenger, the whitening agent, the anti-inflammatory agent, the collagenase activity inhibitor or the collagenase production inhibitor, and the cosmetics or foods containing these in addition to the koji extract, which is an essential component, as well as fat and oil , Waxes, mineral oil, fatty acids or esters thereof, alcohols, polyhydric alcohols, water-soluble polymers such as polysaccharides, polyvinyl polymers and polyacrylic acid polymers, silicones, cationic surfactants, Anionic surfactants, nonionic surfactants, amphiphilic surfactants, organic acids, vitamins, amino acids, nucleic acids, saccharides, extracts derived from plant materials, extracts derived from animal materials, extracts derived from microorganisms , Inorganic salts, inorganic or organic pigments, UV absorbers, antioxidants, anti-inflammatory agents, moisturizers, antibacterial agents, preservatives, enzymes, excipients, thickeners, dyes, fragrances, deodorants or deodorants Agent, Foams, by formulating as an additive various components of seasonings, further possible to provide a cosmetic or food having a function of various. The content in the preparation is not particularly specified, but a concentration range of 0.0001 to 50% by weight is generally used.

  Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto.

Production of cocoon extract using 50% aqueous ethanol solution 100 g of dried cocoon powder was pulverized into a fine powder, added to 1.0 L of 50 w / w% aqueous ethanol solution, and allowed to stand at 25 ° C. for 7 days for extraction. Next, the obtained extract was filtered using filter paper (No. 2 from ADVANTEC), then filtered paper (No. 65 and No. 131 from ADVANTEC) and filter aid (Radiolite P-5 and W- 50), followed by filtration using a φ0.45 μm membrane filter to obtain a 50% ethanol extract (solid content concentration 1.5%) of the cocoon which is the title.

Production of cocoon extract with hot water 50 g of dried cocoon was pulverized into fine powder, added to 1.0 L of purified water, and extracted by heating at 95 ° C. for 3 hours. Next, the obtained extract was filtered using filter paper (No. 2 and No. 65 manufactured by ADVANTEC), and the obtained filtrate was allowed to stand at 4 ° C. for 1 week. Next, the filtrate is filtered using filter paper (No. 65 and No. 131 manufactured by ADVANTEC) and filter aid (Radiolite P-5 and W-50 manufactured by Showa Chemical Industry Co., Ltd.), and then filtered using a φ0.45 μm membrane filter. By performing the above, a hot water extract (solid content concentration: 1.1%) was obtained.

Manufacture of koji extract with 50% 1,3-butylene glycol aqueous solution 50 g of dried koji is pulverized into fine powder and added to 500 mL of 50 w / w% 1,3-butylene glycol aqueous solution at 25 ° C. Extracted by standing for 7 days. Next, the obtained extract was filtered using filter paper (No. 2 from ADVANTEC), then filtered paper (No. 65 and No. 131 from ADVANTEC) and filter aid (Radiolite P-5 and W- 50), followed by filtration using a φ0.45 μm membrane filter to obtain 50% 1,3-butylene glycol extract (solid content concentration 0.9%) of the title candy It was.

Active oxygen scavenging action test In this test, DPPH (diphenylpicrylhydrazyl) radical scavenging method was used. As a sample to be used in this test, a sample obtained by adjusting the koji extract obtained in Example 1 to a solid content concentration of 0.05% and 0.01% using a 50% aqueous ethanol solution, and DL-α-tocopherol as a positive control. A 50% ethanol aqueous solution containing 0.05% and a 50% ethanol aqueous solution containing 0.01% DL-α-tocopherol were used. A DPPH solution, which is a reagent used in this test, was prepared by dissolving DPPH (1,1-Diphenyl-2-picrylhydrazyl, Wako Pure Chemical Industries, Ltd.) in methanol to 6.0 × 10 ⁻⁵ M. A DPPH solution (1.95 mL) and a test sample (0.05 mL) were mixed and reacted at room temperature for 30 minutes under light-shielded conditions. A mixture of 1.95 mL of methanol and 0.05 mL of the test sample under the same conditions was blank, and a mixture of 1.95 mL of DPPH solution and 0.05 mL of 50% aqueous ethanol was used as a control. The absorbance at a wavelength of 515 nm of each mixed solution after the reaction was measured, and the DPPH inhibition rate (%) was calculated from the obtained absorbance value by the following formula.

DPPH radical inhibition rate (%) = [{ABS _C- (ABS _S -ABS _B )} / ABS _C ] × 100
ABS _C : Absorbance in the control
ABS _S : Absorbance in the test sample
ABS _B : Absorbance at blank

  As a result of examining the active oxygen scavenging action of the soot extract, the soot extract of Example 1 (solid content concentration 0.05%) showed a DPPH radical inhibition rate of 100%, and the soot extract of Example 1 In (solid content concentration 0.01%), the inhibition rate was 81.3%. On the other hand, the positive control DL-α-tocopherol solution (concentration 0.05%) showed an inhibition rate of 71.9%, and the DL-α-tocopherol solution (concentration 0.01%) showed an inhibition rate of 34.1%. From the above results, it was found that the koji extract has an excellent active oxygen scavenging action.

Whitening effect test In this test, melanin production inhibitory action was tested as an index of whitening action. As a sample to be used in this test, a sample obtained by adjusting the salmon extract obtained in Example 1 to a solid content concentration of 5 mg / mL using a 50% aqueous ethanol solution, and 5 mg of which whitening action has been confirmed as a positive control. A 50% ethanol aqueous solution containing / mL arbutin and a 50% ethanol aqueous solution as a negative control were used. B16 mouse melanoma cells 3 × 10 ⁴ cells were pre-cultured in 5% FBS-MEM liquid medium (60 mm petri dish) at 37 ° C. under 5% CO ₂ for 24 hours, and then the medium was removed. Next, to the pre-cultured B16 melanoma cells, a 5% FBS-MEM liquid medium and a sample having a concentration of 50 μg / mL with respect to the total amount of the medium were added, and then 72 ° C. under 5% CO _2. Pre-cultured for hours. Next, the medium was removed, and the cells were collected using a Tripsin / EDTA solution. The collected cells were dissolved in 1N NaOH solution containing 10% DMSO to extract melanin in the cells. The optical density (hereinafter referred to as OD475 nm) of the obtained melanin extract at a wavelength of 420 nm was measured, and the obtained numerical value was defined as the amount of melanin produced in this test. The N number was 3, and the average value and standard deviation indicated for each sample were calculated.

  As a result of examining the melanin production inhibitory action of the koji extract, the value of OD475nm in the koji extract of Example 1 is 0.141 ± 0.0081, 0.164 ± 0.0093 in arbutin, and 50% ethanol aqueous solution as a negative control. It was 0.209 ± 0.0091. Since the lowest OD475nm was exhibited in the system to which the koji extract was added, it became clear that the koji extract had an excellent melanin production inhibitory action. In this test, the melanin production inhibitory action was used as an index of the whitening action, so it was found that the koji extract used in the present invention has an excellent whitening action.

Anti-inflammatory test In this test, histamine release inhibitory action was tested as an index of anti-inflammatory action. The test method described in J. Soc. Cosmet. Japan, vol. 25, No. 4, p.246 (1992) was used. As a sample to be used in this test, a sample obtained by adjusting the koji extract obtained in Example 1 to a solid content concentration of 1 mg / mL using a 50% ethanol aqueous solution and a 50% ethanol aqueous solution as a negative control were used. . Removed from the abdominal cavity of rats (4-9 week old Wister male rats, 1 mouse), separated and adjusted by specific gravity centrifugation, 1 x 10 ⁵ cells / mL mast cell suspension 1.2 mL, test sample 0.2 mL Compound 48/80 was added to a final concentration of 1 μg / mL and incubated at 37 ° C. for 15 minutes. Next, the mixed solution containing the mast cell suspension was ice-cooled, and then centrifuged at 3000 rpm for 5 minutes to collect the supernatant. Next, the histamine contained in the supernatant was colored using o-phthaldialdehyde, the absorbance at excitation wavelength 360 nm and fluorescence wavelength 450 nm was measured, and the histamine release inhibition rate (%) was calculated from the following formula. .

Histamine release inhibition rate (%) = [1 − {(ABS _a −ABS _c ) / (ABS _b −ABS _c )}] × 100
ABS _a : Absorbance of a system in which the test sample and compound 48/80 are added to the mast cell suspension
ABS _b : Absorbance of mast cell suspension with compound 48/80 added
ABS _c : Absorbance of the system in which only the test sample is added to the mast cell suspension

  As a result of examining the histamine release inhibitory action of the koji extract, the histamine release inhibition rate in the koji extract of Example 1 was 84%, so that the koji extract has an excellent histamine release inhibitory action. Became clear. In this test, histamine release inhibitory action was used as an index of anti-inflammatory action, so it was found that the koji extract used in the present invention has excellent anti-inflammatory action.

Collagenase activity inhibition test As a sample to be tested in this test, a sample obtained by adjusting the koji extract obtained in Example 1 to a solid content concentration of 0.5% using a 50% ethanol aqueous solution (hereinafter, koji 50% ethanol). A sample (hereinafter referred to as a hot water extract) adjusted to a solid content concentration of 0.5% using the soot extract obtained in Example 2 was used. A 50% aqueous ethanol solution was used as a negative control. Normal human neonatal foreskin dermal fibroblasts (Kurabo) using DMEM (Dulbecco's Modified Eagle's Medium, SIGMA) medium containing 5% FBS (fetal bovine serum, Thermo Trase) and 1% Antibiotis (GIBCO) (Hereinafter, referred to as “fibroblast”) was cultured at 37 ° C. in a 5% CO ₂ atmosphere, the medium was removed, and the fibroblast was washed twice with 2 mL of 25 mM HEPES buffer (manufactured by Dojin Chemical). . A trypsin / EDTA solution (manufactured by Dojindo) adjusted with 25 mM HEPES buffer is added to fibroblasts and reacted for 10 minutes at 37 ° C. in a 5% CO ₂ atmosphere, and 2 mL of 25 mM HEPES buffer containing 10% FBS is added. After 10 minutes, the mixture was centrifuged at 1000 rpm for 5 minutes to recover fibroblasts. Next, 5% FBS and 1% Antibiotic-containing DMEM medium is added to the collected cells to prepare a cell suspension of 1 × 10 ⁵ cells / mL, and this is seeded in a 15 mL, 10 cm culture dish at 37 ° C., The cells were cultured for 24 hours in a 5% CO ₂ atmosphere. After the culture, the medium was removed, and 15 mL of DMEM medium containing 0.02% LAH (lactalbumin hydrolyzate, manufactured by SIGMA) and 10 ng / mL IL-1α (interleukin-1α, Chemicon) was added at 37 ° C., 5 The cells were cultured for 48 hours in a% CO ₂ atmosphere. Next, the culture solution is centrifuged at 10,000 rpm for 10 minutes, 10 μL of 25 mM HEPES buffer containing 1 mg / mL trypsin (manufactured by SIGMA) is added per 100 μL of supernatant, treated at 37 ° C. for 5 minutes, and then 5 mg / mL trypsin. A crude enzyme solution was prepared by adding 10 μL of an inhibitor (GIBCO) -containing 25 mM HEPES buffer to inactivate trypsin. Next, add 50 μL of ice-cooled 1 mg / mL FITC-labeled type 1 collagen (manufactured by Cosmo Bio) and 90 μL of 1.11 mM Tris-HCL buffer (pH 7.5) to a 1.5 mL microtube, mix, and then add 10 μL of the test sample. Added and mixed. Next, 50 μL of a crude enzyme solution (50 μL of purified water in the case of blind test and collagen total measurement) was added to a microtube and heated at 37 ° C. for 2 hours. In the case of measuring the total amount of collagen, it was heated at 95 ° C. for 1 minute. Next, add 200 μL of 10 mM orthophenanthroline-containing 250 mM Tris-HCl buffer to each microtube, stir vigorously for several seconds, let stand at 25 ° C. for 30 minutes, and then centrifuge at 1000 rpm for 15 minutes except for measuring the total amount of collagen. separated. Next, the absorbance of the supernatant was measured using a fluorescence spectrophotometer (Corona Electric Co., Ltd.) at an excitation wavelength of 495 nm and a fluorescence wavelength of 520 nm, and the collagenase activity inhibition rate of the test sample ( %) Was calculated.

Collagenase activity in each system (units / mL) = [(ABS _a −ABS _b ) / (ABS _t −ABS _b )] × 8.33
ABS _a : Absorbance of the system to which the test sample is added
ABS _b : Absorbance of the system used for the blind test
ABS _t : Absorbance of the system used to measure the total amount of collagen

Collagenase activity inhibition rate (%) = [collagenase activity of test sample (units / mL) / collagenase activity of negative control (units / mL)] × 100

  As a result of examining the collagenase activity inhibitory action of the koji extract, the collagenase activity inhibition rate in the koji 50% ethanol extract was 29.5%, and the collagenase activity inhibition rate in the hot water extract was 71.2%. From the above results, it was found that the koji extract has an excellent collagenase activity inhibitory action.

Collagenase production inhibitory action test As a sample to be tested in this test, the koji extract obtained in Example 1 was adjusted to a solid content concentration of 0.1% using purified water and a 30% ethanol aqueous solution. A 30% aqueous ethanol solution was used as a negative control. A 0.1% ascorbic acid solution was used as a positive control. After culturing normal human newborn foreskin skin fibroblasts (manufactured by Kurabo Industries, hereinafter referred to as fibroblasts) at 37 ° C. in a 5% CO ₂ atmosphere using 5% FBS and 1% Antibiotic-containing DMEM medium, The medium was removed and the fibroblasts were washed twice with 2 mL of 25 mM HEPES buffer. Add trypsin / EDTA solution prepared with 25 mM HEPES buffer to fibroblasts, react for 10 minutes at 37 ° C in 5% CO ₂ atmosphere, add 2 mL of 25 mM HEPES buffer containing 10% FBS, and 10 minutes later Then, centrifugation was performed at 1000 rpm for 5 minutes to recover fibroblasts. Next, 5% FBS and 1% Antibiotic-containing DMEM medium is added to the collected cells, and a cell suspension of 6 × 10 ⁴ cells / mL is prepared, seeded on a 24-well plate 1 mL at a time, 37 ° C., 5% CO _2. Culturing was performed for 24 hours under an atmosphere. Next, the medium was removed, 1 mL of 0.02% LAH and 10 ng / mL IL-1α-containing DMEM medium was added, and 10 μL of the test sample was added, followed by culturing at 37 ° C. in a 5% CO ₂ atmosphere for 48 hours. Next, the supernatant was centrifuged at 1000 rpm for 10 minutes, and then the amount of collagenase produced in the supernatant was measured using the MMP-1 Human ELISA System (Amersham Pharmacia Biotech), and the amount of collagenase produced in the negative control (ng / Based on (mL), the collagenase production inhibition rate (%) of the test sample was calculated from the following formula.

Collagenase production inhibition rate (%) = [{collagenase production in negative control (ng / mL) −collagenase production in test sample (ng / mL)} / collagenase production in negative control (ng / mL)] × 100

  As a result of examining the collagenase production inhibitory action of the koji extract, the collagenase production inhibition rate in the koji extract of Example 1 was 85.7%, while in the ascorbic acid which is a positive control, it was 65.6%. From the above results, it was found that the koji extract has an excellent collagenase production inhibitory action.

(Manufacturing examples) Manufacture of various cosmetics or foods Manufacture examples of various cosmetics and foods containing an active oxygen scavenger, whitening agent, anti-inflammatory agent, collagenase activity inhibitor or collagenase production inhibitor containing cocoon extract as an active ingredient However, the present invention is not limited to these.

(Production Example 1) Milk mass%
柿蒂 Ethanol extract 4.0
Squalane 5.0
Olive oil 5.0
Jojoba oil 5.0
Cetyl alcohol 1.5
Glycerol monostearate 2.0
Polyoxyethylene (20) cetyl ether 3.0
Polyoxyethylene (20) soorbitan monooleate 2.0
1,3-butylene glycol 1.0
Glycerin 2.0
P-Hydroxybenzoate ester
Residue with 100 purified water

(Production Example 2) Lotion Toner Mass%
柿蒂 1,3 Butylene glycol extract 3.0
Sodium glutamate 1.0
1,3-butylene glycol 6.0
Glycerin 5.0
Polyethylene glycol 400 3.0
Olive oil 0.5
Polyoxyethylene (20) sorbitan monostearate 1.5
Polyoxyethylene (5) oleyl alcohol ether 0.3
Ethanol 10.0
Perfume Appropriate amount Dye Appropriate amount Phenoxyethanol 0.1
Citric acid 1.0
Sodium citrate 1.2
Residue with 100 purified water

(Production Example 3) Lotion Mass%
Hot water extract 4.5
Glycerin 3.0
1,3-butylene glycol 5.0
Ethanol 15.0
Iron oxide 0.2
Zinc oxide 0.5
Kaolin 2.0
Trehalose 2.0
Dextrin 0.5
Photosensitive element 201 Appropriate amount Photosensitive element 401 Appropriate amount Perfume
Residue with 100 purified water

(Production Example 4) Lotion Mass%
Brine extract 2.0
Sodium citrate 1.0
Dipropylene glycol 5.0
Raffinose 1.0
Ethanol 5.0
Phenoxyethanol 0.2
Pectin 0.01
Xanthan gum 0.05
Acetylglucosamine 0.05
Diisopropylamine dichloroacetic acid 0.2
γ-amino-β-hydroxybutyric acid 0.2
Sodium hyaluronate 0.001
Allantoin 0.2
N-menthyl glyceryl ether 0.2
Dipotassium glycyrrhizinate 0.2
Decarboxycarnosine hydrochloride 0.05
Perfume
Residue with 100 purified water

(Production Example 5) Cream mass%
柿蒂 Ethanol extract 5.0
Stearic acid 2.0
Glycerol monostearate2.0
Cetanol 3.0
Cholesterol 0.5
Vaseline 2.0
Squalane 10.0
Liquid paraffin 10.0
Dimethylpolysiloxane 1.0
Sodium glutamate 2.0
Glycerin 5.0
Dipropylene glycol 3.0
Nicotinamide 1.0
Methyl paraoxybenzoate 0.1
Butyl paraoxybenzoate 0.1
Perfume
Residue with 100 purified water

(Production Example 6) Face wash mass%
柿蒂 50% ethanol aqueous extract 0.5
Sodium aspartate 1.0
Methoxycinnamic acid 1.0
Myristic acid 25.0
Stearic acid 5.0
Tallow fatty acid 5.0
Propylene glycol 10.0
Potassium hydroxide 6.0
Palm oil fatty acid diethanolamide 6.0
Glycerol 1.0
1,3-butylene glycol 1.0
Dipotassium glycyrrhetinate 1.0
Preservative 0.1
Perfume
Residue with 100 purified water

(Production Example 7) Body soap mass%
柿蒂 30% propylene glycol aqueous extract 0.8
Sodium citrate 2.0
N-acetylglutamic acid 1.0
Potassium laurate 15.0
Potassium myristate 5.0
Propylene glycol 5.0
Sorbitol 3.0
Polyethylene powder 0.5
Hydroxypropyl chitosan solution 0.5
Preservative appropriate amount
pH adjuster appropriate amount perfume appropriate amount
Residue with 100 purified water

(Production Example 8) Powder foundation mass%
水溶液 1,3 Butylene glycol aqueous solution extract 3.0
Silicone-treated mica 25.0
Sericite 17.0
Hydrophobic treated silica 20.0
Titanium oxide 20.0
Spherical alumina powder 4.0
Dimethylpolysiloxane 4.0
Methylphenylpolysiloxane 1.0
Vaseline 3.0
Octyl methoxycinnamate 3.0
Sorbitan diisostearate 1.0
Antioxidant Appropriate perfume Appropriate paraoxybenzoic acid
Talc remaining

(Production Example 9) Emulsified liquid foundation mass%
Hot water extract 3.2
Sodium glutamate 1.5
Octamethylcyclotetrasiloxane 20.0
Polyoxyethylene (6) oleyl ether 4.0
Polyoxyethylene hydrogenated castor oil 3.0
Aluminum monostearate 0.01
Liquid paraffin 12.0
Dimethylpolysiloxane 5.0
12-hydroxystearic acid 1.0
Nylon powder 4.5
Black iron oxide 1.5
Yellow iron oxide 3.0
Titanium oxide 5.0
Red iron oxide 2.0
Butyl paraoxybenzoate 0.2
Sorbitol 7.0
Residue with 100 purified water

(Production Example 10) Lipstick mass%
柿蒂 75% aqueous ethanol extract 2.5
Paraffin wax 5.0
Ceresin 5.0
Candelilla wax 2.0
Microcrystalline wax 1.0
Polyethylene wax 1.0
Ethylene propylene copolymer 1.0
Hydrogenated polyisobutene 10.0
Diisostearyl malate 15.0
Isotridecyl isononanoate 12.0
Neopentyl glycol dicaprate8.0
Octyldodecanol 3.0
Squalane 5.0
Phytosteryl oleate 3.0
Methylphenylpolysiloxane 3.0
DL-α-Tocopherol 0.1
Red No. 201 0.4
Red No. 202 0.2
Yellow No.4 0.5
Mica titanium 7.0
Glyceryl trioctanoate

(Production Example 11) Hair shampoo
Brine extract 1.0
Potassium citrate 1.5
N-lauroylmethyl taurine sodium 7.0
Lauroyl sarcosine sodium 6.0
Lauryldimethylbetaine 2.5
Palm oil fatty acid diethanolamide 4.0
Propylene glycol 5.0
2-phenoxyethanol 1.0
Hydroxyethyl cellulose 0.5
Residue with 100 purified water

(Production Example 12) Hair rinse Mass%
柿蒂 50% ethanol aqueous extract 1.5
Sodium pyrrolidonecarboxylate 1.0
Polyoxyethylene / methylpolysiloxane copolymer 2.0
Stearyltrimethylammonium chloride 1.0
Cetostearyl alcohol 3.0
Polyoxyethylene hydrogenated castor oil 1.0
Cationized cellulose 1.0
Isostearyl myristate 1.0
Propylene glycol 5.0
Hydroxybenzophenone 0.1
Perfume
Residue with 100 purified water

(Production Example 13) Hair Treatment Mass%
柿蒂 1,3 Butylene glycol extract 2.0
Behenic acid 5.0
Stearic acid 3.0
Cetyltrimethylammonium chloride 8.5
Diallyl chloride (12-18) dimethylammonium 0.5
Cationized keratin hydrolyzate 0.2
Ceramide 3 0.1
Amino-modified silicone emulsion 0.2
Dipentaerythritol fatty acid ester 0.3
Hydroxymethoxybenzophenone sulfonic acid 0.3
Diethylene glycol monoethyl ether 1.0
Benzyloxyethanol 0.5
Hydroxyethyl cellulose 0.4
Sodium citrate 2.0
Perfume
Residue with 100 purified water

(Production Example 14) Hair nourishing and hair restorer
柿蒂 Ethanol extract 5.0
Sodium glutamate 2.0
Ethanol 60.0
DL-α-tocopherol acetate 0.5
Pepper tincture 0.5
Aloe extract 3.0
Resorcin 0.5
Dipotassium glycyrrhizinate 0.5
Carboxymethylchitin 0.5
Decamethylcyclopentasiloxane 0.1
Arginine 0.25
Glutamine 0.25
Antibacterial agent appropriate amount perfume appropriate amount
Residue with 100 purified water

(Production Example 15) Bath agent mass%
Hot water extract 1.2
Sodium bicarbonate 60.0
Borax 5.0
Sodium hyaluronate 1.0
Hydroxypropyl methylcellulose 1.0
Karin extract 5.0
Antibacterial agent appropriate amount perfume appropriate amount
Residual sodium sulfate 100

(Production Example 16) Ointment Mass%
柿蒂 propylene glycol extract 8.0
l-Menthol 3.0
DL-Camphor 0.5
Propylene glycol 15.0
Carboxyvinyl polymer 1.0
Hydroxyethyl cellulose 0.1
Lauro Macrogol 1.5
Triethanolamine 0.7
Purified water 25.0
Ethanol 100 residue

(Production Example 17) Patch%
Hot water extract 6.0
l-Menthol 3.0
DL-Camphor 1.5
Polyacrylic acid 4.5
Sodium polyacrylate 1.5
Sodium carboxymethylcellulose 4.0
Glycerol 20.0
Sorbitol 5.0
Polyoxyethylene nonylphenyl ether 0.5
Kaolin 5.0
Castor oil 1.0
Dissolving the residue the components to purified water 100, those combined dispersion and mixing, was spread so as to be non-woven polyester fabric 1 m ² per the 1000 g, was prepared patch.

(Production Example 18) Coated preparation A gauze or liniment cloth is impregnated with a prescription liquid in which an ethanol extract, a moisturizer, an anti-inflammatory agent, a cell activator, an antioxidant, etc. are mixed, and attached to the wounded part. Further, an application method in which these are directly applied locally and coated with gauze or the like may be adopted.

(Production Example 19) Capsule
柿蒂 Ethanol extract 20.0g
Magnesium ascorbate 0.5g
DL-α-tocopherol acetate 0.5g
Silicone oil 30.0g
Tween 80 0.05g
Corn starch 48.95g
The above ingredients were mixed uniformly and 200 mg was filled into a gelatin capsule to produce a capsule.

(Production Example 20) Capsule
Hot water extract 15.0g
Starch 60.0g
Mannitol 10.0g
Calcium bicarbonate 5.0g
Polyvinylpyrrolidone 20.0g
The above ingredients were mixed uniformly and 200 mg was filled into a gelatin capsule to produce a capsule.

(Production Example 21) Tablet
柿蒂 5% citric acid aqueous solution extract 100.0mg
Cornstarch 50.0mg
Hydroxypropyl methylcellulose 150.0mg
Carboxymethylchitin 30.0mg
The above ingredients were uniformly mixed and tableted to produce a tablet.

(Production Example 22) Tablet
Brine extract 100.0mg
Corn starch 80.0mg
Methylcellulose 40.0mg
Magnesium stearate 20.0mg
Talc 10.0mg
The above ingredients were uniformly mixed and tableted to produce a tablet.

(Production Example 23) Granule
Hot water extract 50.0mg
Carboxymethylcellulose 180.0mg
Propylene glycol 20.0mg
Sodium alginate 30.0mg
Sucrose 10.0mg
The above ingredients were uniformly mixed and granulated to produce granules.

(Production Example 24) Granule
柿蒂 50% ethanol aqueous extract 60.0mg
Carboxymethylcellulose 180.0mg
Sodium alginate 30.0mg
Erythritol 10.0mg
Citric acid 5.0mg
The above ingredients were uniformly mixed and granulated to produce granules.

(Production Example 25) Syrup
柿蒂 75% ethanol aqueous extract 4.0
Sucrose 5.0
Sorbitol 1.0
Sodium citrate 0.5
Sodium glutamate 0.5
Perfume
The remainder of the purified water 100 was mixed uniformly to produce a syrup.

(Production Example 26) Syrup
Hot water extract 4.0
Ascorbic acid glucoside 0.5
Silk extract 0.2
Coganebana extract 0.2
Sucrose 2.0
Sodium alginate 0.2
Perfume
The remainder of purified water 100 The above ingredients were mixed uniformly to produce a syrup.

Claims (6)

  Reactive oxygen scavenger characterized by using strawberry extract as an active ingredient   A whitening agent characterized by comprising a persimmon extract as an active ingredient   Anti-inflammatory agent characterized by comprising cocoon extract as active ingredient   Collagenase activity inhibitor characterized by comprising persimmon extract as an active ingredient   Collagenase production inhibitor characterized by comprising persimmon extract as an active ingredient A method for preventing or ameliorating photoaging caused by exposure to ultraviolet rays, wherein the extract is taken externally or internally as an active ingredient