WO2008029877A1

WO2008029877A1 - Anti-aging agent, skin-whitening agent, anti-oxidative agent, and anti-inflammatory agent

Info

Publication number: WO2008029877A1
Application number: PCT/JP2007/067402
Authority: WO
Inventors: Hiroko Kikuchi; Masaki Arashima; Hiroko Yoshida
Original assignee: Noevir Co., Ltd.
Priority date: 2006-09-06
Filing date: 2007-09-06
Publication date: 2008-03-13
Also published as: JP2008063266A

Abstract

Disclosed are an anti-aging agent, a skin-whitening agent, an anti-oxidative agent and an anti-inflammatory agent, each comprising a plant Glias nueberthii or an extract thereof as an active ingredient. Further disclosed are various compositions each comprising a plant Glias nueberthii or an extract thereof.

Description

Specification

Anti-aging agents, whitening agents, antioxidants, and anti-inflammatory agents

Technical field

[0001] The present invention relates to an anti-aging agent, a whitening agent, an antioxidant and an anti-inflammatory agent comprising a naturally-derived ingredient as an active ingredient, and more particularly to the use of Glias nueberthii or an extract thereof.

Background art

[0002] As factors of dermatosis such as aging, disease, stress, UV blemishes, stains, skin elasticity reduction, dryness, cell function decline, melanin production and pigmentation by UV rays, dermal matrix components Examples include reduction and degeneration, and cell oxidative damage due to ultraviolet rays. In order to prevent and improve such skin symptoms, various active ingredients have been searched and compounded. In particular, naturally derived ingredients are known to have various pharmacological and cosmetic effects, and so far the application of extracts such as plants and fungi to skin preparations has been studied.

[0003] For example, essence of Ponkan as a cell activator (JP 2001-131045), water and / or organic solvent extract of Hakutsuru Reishi as a whitening agent (JP 2003-89630), antioxidant As an example, an extract of a plant belonging to the genus Sarogase (Japanese Patent Laid-Open No. 10-182413) is known.

Disclosure of the invention

[0004] Thus, various naturally-derived components have been applied so far. There are many natural ingredients that are not yet known for their effects, and have an excellent anti-aging effect, whitening action, antioxidant action, anti-inflammatory action, etc. Development was expected.

The present invention develops naturally-derived ingredients having excellent anti-aging, whitening, antioxidant, and anti-inflammatory effects, and has anti-aging agents, whitening agents, antioxidants, and An object is to provide an anti-inflammatory agent and various compositions.

[0005] As a result of studying various components derived from nature, the present inventors have heretofore confirmed their effects. Ono and Mabo were found to have excellent anti-aging effects, whitening effects, antioxidant effects, and anti-inflammatory effects, and further studies were made to complete the present invention.

That is, according to the first aspect of the present invention, there are provided an anti-aging agent, a whitening agent, an antioxidant, and an anti-inflammatory agent, each of which has an active ingredient such as Glias nueberthii or an extract thereof.

According to a second aspect of the present invention there is provided a composition comprising Glias nueberthii or an extract thereof.

BEST MODE FOR CARRYING OUT THE INVENTION

[0006] Glias nueberthii (scientific name: Hibiscus tiliaceus) is an evergreen tree that belongs to the genus Fuyo of the genus Aoiaceae and is distributed in the subtropical and tropical regions south of the Ryukyu Islands. Ohhamabou or its extract contains an extremely large number of components that cannot be analyzed, and it is speculated that these can act in a comprehensive manner to obtain various effects of the present invention. .

[0007] When using this Ono or Mabo, there are no particular restrictions on the use site, or any site such as flowers, leaves, stems, branches, roots, seeds, bark, sap, pericarp, fruit, buds, etc. You can use force S. A plurality of parts may be used in combination.

It is preferable to pulverize them as they are, and to use the raw material or the dried product, and use an extract from those parts.

[0008] For extraction, any part of the kingfisher may be used, but for simple use, leaves, bark, and the like may be used. At that time, an extract may be obtained using a plurality of parts. In addition, two or more extracts extracted using different solvents may be used in combination.

In the extraction, the plant may be used as it is, but considering the extraction efficiency, it is preferable to carry out the extraction after processing such as shredding, drying, and grinding.

[0009] Extraction methods include room temperature, cooling or warming, extraction by immersing in an arbitrary extraction solvent for a predetermined time, extraction using a distillation method such as steam distillation, or from raw plants Examples of the pressing method include pressing to obtain an extract. Any of these methods alone It is possible to perform extraction with a combination of two or more. Alternatively, extraction can be performed using a supercritical fluid or a subcritical fluid. In order to increase the extraction efficiency, the mixture may be stirred or homogenized in an extraction solvent. It is also possible to extract under a caloric pressure using an autoclave.

[0010] The extraction temperature is suitably about 5 ° C to the boiling point of the extraction solvent. The extraction time varies depending on the type of extraction solvent and the extraction temperature, but it is appropriate to set it for about 1 hour to 14 days.

The ratio of the plant and the solvent during the extraction is not particularly limited, but is preferably 0.5 to 5 times the solvent for the plant 1; Preferably 100 times by mass.

[0011] Extraction solvents include water, lower alcohols such as methanol, ethanol, propanol and isopropanol; polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, dipropylene glycol and glycerin; ethyl ether and propyl ether Solvents such as ethers such as butyl acetate, butyl acetate, ethyl acetate, and the like; ketones such as acetone, ethylmethylol ketone, and the like can be used. These may be used alone or in combination of any two or more. Saline, phosphate buffer, phosphate buffered saline and the like may be used. In addition, one or more supercritical liquids and subcritical liquids such as water, carbon dioxide, ethylene, propylene, ethanol, methanol, and ammonia may be used.

[0012] Extracts of ono and mabo using the above solvents may be used as they are, may be left standing for a period of time and aged, or the concentrated and dried solids may be reconstituted in water or a polar solvent. It can also be used after being dissolved. Alternatively, it may be used after performing purification treatment such as decolorization, deodorization, desalting, etc., or fractionation treatment by column chromatography or the like within a range not impairing these physiological functions. The above-mentioned extract of wolfberry, its processed products and fractions can be freeze-dried after each processing and fractionation and dissolved in a solvent before use. It can also be used by being encapsulated in vesicles such as liposomes or microcapsules.

[0013] ono, mabo or an extract thereof has excellent anti-aging, whitening, antioxidant, and anti-inflammatory effects, and is preferably used as an anti-aging agent, whitening agent, antioxidant, and anti-inflammatory agent. Use with power S.

Each of these agents is not limited at all in terms of its form and the presence or absence of other ingredients as long as it contains honono, mabo or an extract thereof as an active ingredient. As for the form, any form such as liquid, paste, gel, solid, powder, etc. can be selected according to its use, etc., and the vehicle (excipient) necessary for obtaining the form, Solvents and other common additives (antioxidants, colorants, dispersants, etc.) can optionally be included.

[0014] The amount of the active ingredient, Ono, Mabou or its extract, in each agent is the total amount in terms of force S, effect, and stability that can be adjusted according to the type and purpose of use of the agent. to be, 0. 00001～ in terms of solid content; preferably from 100 mass ^_0/0 force Mashigu <is 0.00; is ~ 50 mass%!.

[0015] An anti-aging agent comprising ceramata or an extract thereof as an active ingredient has an excellent cell activation effect, collagen production action, and aromatase activity promotion action, and exhibits an excellent effect in improving the prevention of aging symptoms. To do. Aromatase is an enzyme that works when producing estrogen. If the production of estrogen is promoted by the action of promoting aromatase activity, the effect of female hormones on skin can be expected.

[0016] A whitening agent comprising ono, mabo or an extract thereof as an active ingredient has an excellent melanin production inhibitory effect and tyrosinase activity inhibitory effect, and prevents and improves pigmentation, stains, freckles, etc. Exhibits excellent whitening effect.

[0017] Antioxidants containing ceramia or its extract as an active ingredient have an excellent free radical scavenging effect and superoxide anion scavenging effect, which prevents skin photoaging and the like. Demonstrates oxidizing action.

[0018] An anti-inflammatory agent comprising wolfberry or its extract as an active ingredient has excellent hyaluronidase inhibitory effect and phospholipase A2 activity inhibitory effect, and exhibits excellent anti-inflammatory action by suppressing skin inflammation. .

[0019] Each of these agents can be used for hair or the like as well as applied externally to the skin, and can be applied to various compositions such as an external composition and an oral composition. Here, the composition for external use includes cosmetics, external preparations for skin, quasi-drugs, external medicines, etc. It is not limited to any one category, but means all compositions applied externally to the skin or hair. The term “oral composition” means any composition that can be taken orally, regardless of the type of drug, food, beverage, etc.

[0020] The dosage form of the composition for external use is arbitrary, and can be provided as, for example, a solubilization system such as lotion, a dispersion system such as calamine mouth lotion, or an emulsification system such as cream or emulsion. In addition, it can be provided in various dosage forms such as aerosol forms, ointments, and poultices filled with propellants.

Specifically, various cosmetics such as emulsions, creams, lotions, lotions, packs, cosmetic liquids, cleaning agents, lipsticks, makeup cosmetics, foundations, etc .; liquids, ointments, powders, granules, aerosols, patches Examples include various forms of quasi-drugs such as cataplasms and cataplasms for external use.

[0021] The form of the oral composition is also arbitrary, and is in a liquid form such as a liquid, syrup, or extract, or a solid such as a granule, tablet, powder, capsule, glaze, or jelly, It can be processed and used in various forms such as gummi and gum, and is not particularly limited.

Specific examples include general foods including beverages, health foods (supplements), functional foods, nutritional supplements, oral medicines, and quasi drugs.

[0022] By adding ono, mabo or an extract thereof to an external preparation for skin including, for example, cosmetics, external medicines, quasi-drugs, skin wrinkles, tarmi, harsh skin, Therefore, it is possible to obtain a composition for external use that exhibits an excellent effect in preventing and improving various skin symptoms, and it can be used as a skin external preparation for moisturizing or a skin external preparation for whitening. In particular, it is preferable to use it as a skin external preparation for improving aging prevention, which includes Ohhamabou or its extract.

In addition, ono, mabo or its extract should be used for food and drink, health food (supplements), quasi-drugs, functional foods, etc. for the purpose of beauty such as whitening, health maintenance or nutritional supplementation. It ’s a monkey.

[0023] The composition such as an external composition or an oral composition includes, in addition to ohahabo or an extract thereof, a normal skin cosmetic, a hair cosmetic, a quasi-drug, depending on its use and necessity, Any component used in pharmaceutical preparations is included. As such an ingredient , Water, oil (oil component), alcohols, excipients, binders, extenders, disintegrants, corrigents, pigments, colorants, emulsifiers, solubilizers, dispersants, gelling agents, plasticizers, washing Agent, UV absorber, thickener, pH adjuster, buffer, surfactant, lubricant, chelating agent, drug (medicine component), fragrance, resin, coating agent, antibacterial and antifungal agent, antiseptic Agents, preservatives, antioxidants, pH adjusters and the like. In addition, other anti-aging agents, whitening agents, antioxidants, anti-inflammatory agents, or plants other than wolves or extracts thereof can be used as long as the effects of the present invention are not impaired.

[0024] In the case of foods and drinks including ono, mabo, or an extract thereof, the combination with various components used in foods is not particularly limited.

In other words, the dosage form of food is arbitrary, and can be provided in various dosage forms such as powders, granules, capsules, liquids, etc. Oil components, moisturizers, powders, emulsifiers, solubilizers, thickeners, drugs, fragrances, antifungal agents, alcohols, sugar, condensed milk, flour, salt, glucose, chicken eggs, butter, marga Phosphorus, chickenpox, calcium, iron, seasonings, spices, vitamin A and their derivatives, strength rotenoids, riboflavin and its derivatives, vitamin B and its salts or derivatives, ascorbic acid and its derivatives, cobalamins, vitamin E And their derivatives, vitamins, adenosine and its derivatives, flavonoids and tannins. Furthermore, other anti-aging agents, whitening agents, antioxidants, or anti-inflammatory agents can be used in combination as long as the effects of the present invention are not impaired.

[0025] The blending amount of the orphan or its extract in a composition such as an external composition or an oral composition can be adjusted according to the kind of composition, purpose of use, etc. from the total amount, 0.1 in terms of solid content 00001-50. it is preferably 0 mass%, more preferably, 0.5 0001- 25. 0 mass ^_0/0. Further, 0.0010 to 10% by mass is more preferable 0.000;! To 5% by mass, more preferably 0.00;! To 5% by mass, and still more preferably 0. 0;! To 5% by mass, particularly preferably 0 .;! To 5% by mass.

Example

[0026] The following is a preparation example of the extract of the wolfberry, a test for evaluating each action, and a topical skin application. The ability to explain the formulation examples as agents and the like in more detail S, and the technical scope of the present invention is not limited to these.

<0027 Extraction Method 1: Ethanol Extract>

Ono and burdock leaves or bark were dried and crushed, and 50% ethanol by mass of 20 times the mass of the sample was added, followed by extraction with stirring at room temperature for 2 hours. The obtained extract was filtered to remove insolubles, concentrated under reduced pressure, and lyophilized to obtain an extract.

[0028] <Extraction method 2: hot water extract>

Ono, burdock leaves or bark were dried and pulverized, purified water of 20 times the sample mass was added, and extracted by heating to 120 ° C for 20 minutes by autoclave. From the obtained extract, insolubles were removed by suction filtration while maintaining a high temperature state, and then freeze-dried to obtain an extract.

[0029] <Evaluation of human dermal fibroblast activation effect>

The dermis fibroblast activation action was evaluated using the ono and burdock leaf ethanol extracts obtained in Extraction Method 1 as follows.

Normal human dermal fibroblasts manufactured by Kurashiki Boseki Co., Ltd. were seeded in a 96-well microphone mouth plate so that 2.0 × 10 ⁴ cells per well. The seeding medium used was Dulbecco's modified Eagle medium (DMEM) supplemented with 1% by weight urchin fetal serum (FBS). After culturing for 24 hours, the sample culture medium was adjusted to each sample concentration shown in Table 1 with 1 mass ° / 83-added DMEM medium, and further cultured for 48 hours. After removing the supernatant, the medium was replaced with a medium containing 400 µg / ml of 3- (4,5-dimethylthio-2-thiazolyl) -2,5-diphenyltetrazolium bromide (MTT reagent). Cultured for 2 hours. Then, the formazan produced by the opening of the tetrazolium ring was extracted with 2-propanol, and the absorbance at 55 Onm was measured with a microplate reader. At the same time, the absorbance at 650 nm was measured as turbidity, and the cell activation effect was evaluated by the difference between the two measured values.

The obtained results are shown in Table 1 as relative values when the cell activation effect in the control without addition of the sample is taken as 100.

[0030] [Table 1] Sample addition concentration (mg / ml)% of control

Con 卜 Ronore 100

0. 03 1 11

0. 06 1 19

0. 13 130

0. 25 161

0. 5 241

[0031] As is clear from Table 1, a significant dermal fibroblast activation effect was observed in the medium supplemented with the wolfberry extract.

[0032] <Evaluation of human epidermal cell activation effect>

Using the hot water leaf extract obtained by the above extraction method 2, the skin cell activation effect was evaluated as follows.

Human epidermal non-keratinized cells were seeded in a 96-well microphone mouth plate so that there were 2.0 × 10 ⁴ cells per well. The seeding medium used was Dulbecco's modified Eagle medium (DMEM) supplemented with 5% by weight urine fetal serum (FBS). After culturing for 24 hours, the culture medium was replaced with a sample culture solution adjusted to the concentration of each sample shown in Table 2 using 5 mass ° / ^ 8 S-added DMEM medium, and further cultured for ²⁴ hours. After removing the supernatant, the medium was replaced with a medium containing 100 gZml of MTT reagent and cultured for about 2 hours. Subsequently, fonolemazan produced by the opening of the tetrazolium ring was extracted with 2-propanol, and the absorbance at 550 nm was measured with a microplate reader. At the same time, the absorbance at 650 nm was measured as turbidity, and the cell activation effect was evaluated by the difference between the two measured values.

The obtained results are shown in Table 2 in terms of relative values when the cell activation effect in the control with no sample added is taken as 100.

[0033] [Table 2]

[0034] As is clear from Table 2, a significant epidermal cell activation effect was observed in the medium supplemented with the wolfberry extract.

<Evaluation of human dermal fibroblast collagen production> Using the ono and burdock bark ethanol extracts obtained by extraction method 1 above, the dermal fibroblast collagen production action was evaluated as follows.

Normal human dermal fibroblasts were seeded on a 96-well microphone plate at 2.0 × 10 ⁴ per well. The seeding medium used was Dulbecco's modified Eagle medium (DMEM) supplemented with 5% by weight urine fetal serum (FBS). After culturing for 24 hours, the sample culture solution was adjusted to the sample concentration shown in Table 3 with DMEM medium supplemented with 0.5 mass% FBS, and further cultured for 24 hours.

The ELISA method was used to quantify type I collagen secreted into the culture supernatant, and finally 2, 2 'azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) against the labeled peroxidase. After adding and reacting with hydrogen peroxide, absorbance at 405 nm was measured with a microplate reader.

The amount of protein was measured by PIERCE BCA Protein Assay Kit, and the amount of type I collagen produced per unit protein was determined.

The obtained results are shown in Table 3 as relative values when the amount of type I collagen produced per unit protein in the control with no sample added is defined as 100.

[Table 3]

[0037] As is clear from Table 3, a significant dermal fibroblast collagen production effect was observed in the medium supplemented with ono and mabo extracts.

[0038] <Evaluation of collagen production by human epidermal cells>

Using the ono and burdock bark ethanol extract obtained by extraction method 1 above, the epidermal cell collagen producing action was evaluated as follows.

Human epidermal non-keratinized cells were seeded in a 96-well microphone mouth plate so that there were 2.0 × 10 ⁴ cells per well. The seeding medium used was Danolebecko modified ignore medium (DMEM) supplemented with 5% by weight urchin fetal serum (FBS). After incubation for 24 hours, replace with sample culture solution adjusted to each sample concentration shown in Table 4 with DMEM medium containing 5 mass ° / ^ 8 S And further cultured for 5 days.

For the quantification of type IV collagen secreted into the culture supernatant, sandwich ELISA using a monoclonal antibody against IV collagen (recognition site: _α2 chain) and biotinylated polyclonal antibody was used, and avidinized horseradish peru After adding xidase, color was developed with 3,3 ′, 5,5′-tetramethylbenzidine, and the absorbance at 650 nm was measured with a microplate reader.

The amount of protein was measured using the BCA Protein Assay Kit manufactured by PIERCE, and the amount of type IV collagen produced per unit protein was determined.

The obtained results are shown in Table 4 as relative values when the amount of type IV collagen production per unit protein amount in the control with no sample added is defined as 100.

[Table 4]

[0040] As is clear from Table 4, a significant epidermal cell collagen production effect was observed in the medium supplemented with Ono and Mabo extracts.

[0041] <Evaluation of aromatase activity promoting action>

Using the ono and burdock ethanol extracts obtained by extraction method 1 above, the aromatase activity promoting action was evaluated as follows.

Sample solutions prepared for each sample concentration shown in Table 5 41, NADP +, MgCl, dull

2-course 6-phosphate, glucose 6-phosphate dehydrogenase, and control insect cell membrane protein mixed solution (CPY19 / MFC High Throughput Inhibitor Screening Kit, manufactured by BD Biosciences) 96 1 Add and warm to 37 ° C for 10 minutes. 15 nM CPY19 (Alomata 1ze), 50 M 7 methoxy-1 4 trifluoromethylcoumarin (substrate) solution 100 1 were added, and the mixture was heated to 37 ° C. for 30 minutes. lOOmM Tris base 75 1 was added to stop the reaction. Fluorescence measurement was performed at an excitation wavelength of 409 nm and an emission wavelength of 530 nm. Since 7-methoxy-4-trifluoromethylcoumarin was decomposed by CYP19 and 7-hydroxy-4-trifluoromethylcoumarin was produced to produce fluorescence, the ability to promote aromatase activity was quantified by fluorescence measurement.

The obtained results are shown in Table 5 as relative values when the aromatase activity promoting action in the control with no sample added is taken as 100.

[Table 5]

P 0.01, *: 0.01 P 0.05

[0043] As is clear from Table 5, a significant aromatase activity-promoting action was observed in the wolfberry extract.

[0044] <Evaluation of human epidermal melanocyte tyrosinase activity inhibitory action>

Using the ono and burdock leaf ethanol extracts obtained by Extraction Method 1 above, the epidermal melanocyte tyrosinase activity inhibitory action was evaluated as follows.

Normal human epidermal melanocytes were seeded on a 96-well microphone mouth plate at 3.0 × 10 ⁴ cells per well. As the seeding medium, Medium 154S manufactured by Kurashiki Boseki Co., Ltd. was used. 24 After culturing for 4 hours, the medium was replaced with a sampnore medium adjusted to each sample concentration shown in Table 6 using Medium 154S, and further cultured for 48 hours. Next, the cells were completely lysed by exchanging with phosphate buffer 751 containing 1% by mass 1¾101-X, and 501 of which was used as a crude enzyme solution. To the crude enzyme solution, 0.05% by mass as a substrate and 50 / dopa-containing phosphate buffer was added, and the mixture was allowed to stand at 37 ° C. for 2 hours. The absorbance at 405 nm was measured immediately after substrate addition and at the end of the reaction with a microplate reader, and each measured value was introduced into the following equation to determine the amount of produced dopamelanin.

Dono melanin production =

{(After reaction, 405 — value—Before reaction, 405 stomach value) — 2. 166} / 5. 238

In addition, the amount of protein was measured using BCA Protein Assay Kit manufactured by PIERCE. The amount of melanin produced per protein amount was determined. The case where no sample was added was used as a control. The results are shown in Table 6.

[0045] [Table 6]

[0046] As is clear from Table 6, in the medium supplemented with Ono and Mabou extracts, a decrease in melanin production was observed and an excellent inhibitory effect on tyrosinase activity was observed.

[0047] <Evaluation of B16 mouse melanoma cell melanin production inhibitory effect>

Using the ono and burdock bark ethanol extracts obtained by extraction method 1 above, the melanin production inhibitory action was evaluated as follows.

B16 mouse melanoma cells (B16F0 cells) were seeded in 90 mm dishes so that there were 1 · 8 × 10 ⁴ cells per dish. The seeding medium used was Dulbecco's modified Eagle medium (DMEM) supplemented with 5% by weight urine fetal serum (FBS). After culturing for 24 hours, the sample culture solution was adjusted to the concentration of each sample shown in Table 7 using 5-medium i% FBS-added DMEM medium, and further cultured for 5 days. After completion of the culture, the cells were detached by trypsin treatment, transferred to a 1.5 ml microtube, and centrifuged to obtain a cell precipitate. The color of the resulting precipitate was visually judged based on the following criteria for blackening. The evaluation is based on a 5-step evaluation, 5 mass ° / (^ 83 added DME M medium with no sample added to the negative control (judgment 5), 5% FBS-added DMEM medium containing 50 mM sodium lactate in the positive control (judgment 1) Was used.

In addition, in order to evaluate the amount of melanin produced, the cell lysing agent Solvable (Perkin Elma Japan Co., Ltd.) was added to the precipitate, boiled, returned to room temperature, and spectrophotometer (manufactured by Hitachi High-Technologies Corporation) Absorbance at 500 nm was measured with a total of U-3010).

The results obtained are shown in Table 7.

[0048] <Criteria for visual evaluation> Judgment Precipitation color

1 Same as positive control (almost white)

2 Slightly darker than the positive control (light! /, Brown)

3 Between the positive control and negative control (brown) 4 Slightly less blackening than the negative control (blackish brown) 5 Same level as the negative control (almost black)

[0049] [Table 7]

[0050] As can be seen from Table 7, a significant melanin production-suppressing effect was observed in the medium supplemented with Ono and Mabo extracts.

[0051] <Evaluation of antioxidant effect by scavenging DPPH radicals>

Using the ono and burdock bark hot water extracts obtained by Extraction Method 2 above, the antioxidant effect by scavenging DΡΡΗ radicals was evaluated as follows.

Using 50% by mass ethanol, the sample solution was adjusted so that each sample concentration shown in Table 8 was obtained, and 100 aliquots were added to a 96-well microplate. Thereto, 0.2 mM of 1,1-diphenyl-2-picrylhydrazyl (DPPH) ethanol solution was added in a volume of 100 to 1 and mixed well, and allowed to stand at room temperature in the dark for 24 hours. Finally, the absorbance at 516 nm derived from DPPH radical was measured.

The extinction rate of DPPH radical was calculated from the following equation, where (A) is the absorbance when the sample is not applied and (B) is the absorbance when the sample is added.

Radical scavenging rate = {1 (B) / (A)} X 100

The results are shown in Table 8.

[0052] [Table 8] Sample addition concentration ( _mg / ml) Scavenging Act activity (%)

1 85

10 96 [0053] As can be seen from Table 8, the extract of Ohamabou had an excellent DPPH radical scavenging effect.

[0054] <SOD-like activity evaluation (evaluation of superoxide anion scavenging ability)>

The SOD-like activity was evaluated using the ono and burdock leaf ethanol extracts obtained by Extraction Method 1 as follows.

0. HANK 'S (+) solution 7 511 1 containing 25 mM WST—1 and ImM Hypoxanthine was added to the sample solution 25 5 1 prepared with the HANK' S (+) solution at each sample concentration shown in Table 9. did. Further, xanthine oxidase 25 ^ 1 (0.00 0075 Units) was added and reacted at 37 ° C for 15 minutes, and then the absorbance at 450 nm was measured. When the absorbance when only the HANK 'S (+) solution is added instead of the sample solution is (A) and the absorbance when the sample solution is added is (B), the superoxide anion elimination rate is expressed by the following equation: Defined.

Erasure rate (%) = {1— (B) / (A)} X 100

The results obtained are shown in Table 9.

[0055] [Table 9]

[0056] As is clear from Table 9, an excellent superoxide anion scavenging effect was observed in the wolfberry extract.

[0057] <Evaluation of phospholipase A (PLA) inhibitory action>

twenty two

Using the ono and burdock bark ethanol extracts obtained by extraction method 1 above, the phospholipase A (PLA) inhibitory action was evaluated as follows.

twenty two

Phospholipase A (PLA) adjusted to a final concentration of 60 ng / ml and the concentrations shown in Table 10

twenty two

DTNB (5,5-dithiobis (2-nitrobenzoic acid) adjusted to 10 mM, mixed for 10 minutes at room temperature, and 1.66 mM diheptanoylthio-PC as substrate (Diheptanoyl Thio-PC) was added and allowed to react at room temperature for 45 minutes, and then the absorbance at 414 nm was measured, and only the buffer was added instead of the PLA solution. The absorbance when added was measured to determine the difference between the two measurements. When the value of the control with no sample added is (A) and the value at the time of sample addition is (B), the PLA enzyme inhibitory action is defined as

2

I will.

Erasure rate (%) = {1— (B) / (A)} X 100

The results obtained are shown in Table 10.

[Table 10]

[0059] As can be seen from Table 10, the Oohambobo extract has an excellent PLA enzyme inhibitory action.

2

Admitted.

[0060] <Evaluation of hyaluronidase inhibitory action>

The hyonuronidase inhibitory action was evaluated using the ono and burdock leaf ethanol extracts obtained by Extraction Method 1 as follows.

A commercially available hyaluronic acid potassium salt (derived from human umbilical cord) was dissolved in 0.1 M phosphate buffer (ρΗ7.0) so as to be 0.9 mg / ml to obtain a substrate solution. A commercially available hyaluronidase (derived from urchin testis) was dissolved in 0.1 M phosphate buffer (pH 7.0) so as to be 5,300 unit / ml to obtain an enzyme solution. The enzyme solution was prepared at the time of use.

A sample solution (0.1 ml) and an enzyme solution (0.03 ml) prepared to each sample concentration shown in Table 11 with a buffer solution were placed in a test tube and reacted at 37 ° C for 20 minutes. Next, 0.06 ml of an activator was added and reacted at 37 ° C for 20 minutes. Further, 0.15 ml of the substrate solution was added and reacted at 37 ° C for 1 hour. Stop the reaction by adding 0.06 ml of 4N NaOH aqueous solution, then immediately ice-cool, add 0.06 ml of borate buffer (pH 9.1), boil for 3 minutes, and further ice-cool. did. Add 20 ml of p-DABA (p-dimethylaminobenzaldehyde) solution, react at 37 ° C for 20 minutes, transfer from each tube to a 96-well microphone mouthplate, and use a microplate reader. Absorbance at 585 nm was measured. For control, a buffer solution with no sample added was used. When the activity of hyaluronidase is inhibited, the degradation product N-acetylyldarcosamine (GlcNAc) decreases, and the absorbance due to p-DABA decreases. The hyaluronidase inhibitory action is defined by the following formula. Inhibition rate (%)

= (Control Absorbance-Sampnore Absorbance) z Control Absorbance Table 11 shows the results.

[0061] [Table 11]

[0062] As is clear from Table 11, the extract of Ohamabou had an excellent inhibitory effect on hyaluronidase activity.

Then, the formulation example which implemented this invention is shown.

[Example 1] Emulsion

[Table A]

(1) Screening 0.0 (mass%)

(2) Methyl fuel polysiloxane 4.0

(3) Hydrogenated palm kernel oil 0.5 (4) Hydrogenated soybean phospholipid 0.1

(5) Polyoxyethylene sorbitan monostearate E. o.)

13

(6) Sorbitan monostearate1.0

(7) Glycerin 4.0

(8) Methyl paraoxybenzoate 0.1

(9) Carboxyvinyl polymer 0.15

(10) The balance of purified water 1 0 0

(11) Arginine (1 mass% aqueous solution) 2 0. 0

(12) Yellowtail extract (Extraction method 1, leaves) 3.0 Production method: Heat-dissolve the oil phase components (1) to (6) at 80 ° C. On the other hand, the aqueous phase components (7) to (10) are heated and dissolved at 80 ° C. The oil phase component is added to this while stirring and uniformly emulsified with a homogenizer. After emulsification, start cooling and add (11) and (12) sequentially and mix evenly.

[0064] [Example 2] Lotion

[Table B] (1) Ethanonore 1 5. 0 (mass%)

(2) Polyoxyethylene (4 0 E. O.) Hardened castor oil 7 0.3

(3) Fragrance 0. 1

(4) The balance of purified water 1 0 0

(5) Chenic acid 0 · 0 2

(6) Sodium taenoate 0.1

(7) Glycerin 1.0

(8) Hydrochestino Resenorelose 0.1

(9) Wolves' Extract (Extraction method 2, leaves) 5.0 Manufacturing method: Dissolve (2) and (3) in (1). After dissolution, add (4) to (8) in sequence, and then stir well. Add (9) and mix uniformly.

[Example 3] Cream

[Table C]

(1) Scorran 1 0.0 (mass

(2) Stearic acid 2.0

(3) Hydrogenated palm kernel oil 0.5

(4) Hydrogenated soybean phospholipid 0.1

(5) Cetanol 3.6

(6) Lipophilic glycerin monostearate2.0

(7) Glycerin 1 0. 0

(8) Methyl paraoxybenzoate 0.1

(9) Arginine (20 mass% aqueous solution) 1 5. 0

(10) The balance of purified water 1 0 0

(11) Carboxybule polymer (1% by weight aqueous solution) 1 5. 0

(12) Wolves' Extract (Extraction Method 1, Vegetables) 5.0 Manufacturing method: The oil phase components (1) to (6) are heated and dissolved at 80 ° C. On the other hand, the aqueous phase components (7) to (10) are heated and dissolved at 80 ° C. The oil phase component is added to this while stirring and uniformly emulsified with a homogenizer. After emulsification, add (11), start cooling, add (12) at 40 ° C and mix uniformly.

[Example 4] Cosmetic liquid

[Table D] Purified water 0 0 Remainder (mass%) Glycerin 0 0

Sucrose fatty acid esters 3

(4 Carboxyvinyl polymer (1 mass / ₀ aqueous solution) 7 5 (5 Sodium alginate (1 mass% aqueous solution) 5 0 (6 Polyglyceryl monolaurate 0 (7 Macademi Anatsu oil fatty acid phytosteryl 30 (8 N- Lauroyl-L-glutamate di (phytosteryl. 2-octyldodecyl)

2. 0

(9) Hardened palm oil 2.0

(10) Sukuran (from olive) 1.0

(11) Behe-Norecore 0. 7 5

(12) Mitsuru 1.0

(13) Hohono oil 1. 0

(14) 1, 3-Pitreneglycol 1 0. 0

(15) L_Arginine (10 mass% aqueous solution) 2.0

(16) Yellowtail extract (extraction method 2, leaves) 5.0 Manufacturing method: Mix the aqueous phase components (1) to (6) and dissolve at 75 ° C with heating. On the other hand, the oil phase components (7) to (14) are mixed and dissolved by heating at 75 ° C. Next, the oil phase component is added to the aqueous phase component and preliminary emulsification is performed, followed by uniform emulsification with a homomixer. Start cooling after emulsification and add (15) at 50 ° C. Cool to 40 ° C, add (16), and mix evenly.

[0067] [Example 5] Aqueous jewel

[Table E]

(1) Carboxyvinyl polymer 0.5 (mass./.)

(2) The balance of 100 purified water

(3) Sodium hydroxide (10 mass% aqueous solution) 0.5

(4) Ethanol 10.0

(5) Methyl paraoxybenzoate 0.1

(6) Fragrance 0.1

(7) Wolves extract (extraction method 2, leaves) 3.0

(8) Polyoxyethylene (60 EO) hardened coconut oil = 7 coconut oil 1.0 Manufacturing method: Add (1) to (2) and stir uniformly, then add (3). After stirring uniformly Add (5) dissolved in (4) in advance, and after stirring uniformly, add (6) to (8) previously mixed and stir and mix uniformly.

[0068] [Example 6] Cleansing fee

[Table F] (1) Screen 8 1.0 (mass ⁰ /.)

(2) Polyoxyethylene glyceryl isostearate 1 5.0

(3) The balance of purified water 1 0 0

(4) Wolves extract (Extraction method 1, bark) 4.0 Manufacturing method: Dissolve (1) and (2) uniformly. (3) and (4) are sequentially added to this and mixed uniformly.

[Example 7] Face washing foam

[Table G]

(1) Stearic acid 16 (mass%)

(2) Myristic acid 16

(3) Lipophilic glycerin monostearate 2

(4) Glycerin 2 5

(5) Sodium hydroxide

(6) Coconut oil fatty acid amidopropyrubetaine

(7) The balance of purified water 00

(8) Yellowtail extract (Extraction method 2, bark) 5.0 Manufacturing method: The oil phase component of (14) is heated and dissolved at 80 ° C. On the other hand, the water phase components (5) to (7) are heated and dissolved at 80 ° C and mixed with the oil phase components uniformly. Start cooling, add (8) at 40 ° C, and mix uniformly.

[Example 8] Makeup base cream

[Table H]

(1) Screen 10.2 (mass. / ₀ )

(2) Cetano 2.0

(3) Glycerol 1-ethylhexanoate 2.5

(4) Lipophilic glyceryl monostearate 1.0

(5) Propyrene glycol 1 1. 0

(6) Sucrose fatty acid ester 1.3

(7) The balance of purified water 1 0 0

(8) Titanium oxide 1.0

(9) Bengala 0.1

(10) Yellow iron oxide 0.4

(11) Fragrance 0.1

(12) Yellowtail extract (extraction method 2, leaf) 3.0 Manufacturing method: Mix the oil phase components of (1 4) and dissolve at 75 ° C with heating. On the other hand, (5 7) aqueous phase component is mixed and dissolved by heating at 75 ° C, and (8) 10) pigment is added to this and dispersed uniformly with a homomixer. The oil phase component is added to the water phase component and emulsified with a homomixer. Start cooling after emulsification, add ingredients (11) and (12) at 40 ° C and mix evenly. [Example 9] Emulsion foundation

[Table I]

1) Methyl ^^ polysiloxane 2.0 (mass%)

2) Skullang 5.0

3) Otachidodecyl myristate 5.0

4) Cetano 1.0

5) Polyoxyethylene (20 E. O.) sorbitan monostearate

13

6) Sorbitan monostearate0.7

7) 1,3-Butyleneglycol 8.0

8) Xanthan gum 0.1

9) Methyl paraoxybenzoate 0.1

10) The balance of purified water 1 0 0

11) Titanium oxide 9.0

12) Talc 7.4

13) Bengala 0.5

14) Yellow iron oxide 1.1

15) Black iron oxide 0.1

16) Perfume 0.1

17) Wolves extract (extraction method 1, leaves) 4.0 Manufacturing method: Mix the oil phase components of (16) and dissolve at 75 ° C with heating. On the other hand, the water phase components (7) to (10) are mixed, dissolved by heating at 75 ° C, and the pigments (11) to 15) are added to this and uniformly dispersed with a homomixer. Add oil phase ingredients and emulsify. Cooling is started after emulsification, and components (16) and (17) are added sequentially at 40 ° C and mixed uniformly.

[Example 10] Water-in-oil emollient cream

[Table J]

(1 Flowing ballaffin 3 4 0 (mass%)

(2 Microcrystalline wax 2 0

(3 Wasselin 5 0

(4 Diglycerinoleate 5 0

(5 Sodium chloride 3

(6 Lithium chloride 0

(7 Propylene glycol 3 0

(8 1,3-butylene glycol 5 0

(9) Methyl paraoxybenzoate 0

(10) Acer fountain extract (Extraction methods 1, 30

(11) The balance of purified water 00

(12) Fragrance 0.1 Production method: Dissolve (5) and (6) in a part of (11) to 50 ° C, heat to 50 ° C and gradually add to (4) while stirring. After mixing this, heat-dissolve at 70 ° C to uniformly disperse into (13). Scatter. To this, add (7) to (10) with the remainder of (11) dissolved at 70 ° C while stirring, and emulsify with a homomixer. Start cooling after emulsification, add (12) at 40 ° C, and mix uniformly.

[Example 11] Pack

[Table K]

(1) The balance (mass%) of purified water 100

(2) Polybulu alcohol 1 2.0

(3) Ethanol 1 7.0

(4) Glycerin 9.0

(5) Polyethylene glycol (average molecular weight 1 0 0 0) 2.0

(6) Euryoptera extract (extraction method 2, leaves) 5.0

(7) Fragrance 0.1 Production method: (2) and (3) are mixed, heated to 80 ° C, and then dissolved in (1) heated to 80 ° C. After evenly dissolving, add (4) and (5) and start cooling with stirring. Cool to 40 ° C, add (6) and (7), and mix evenly.

[0074] [Example 12] Bath salt

L]

(1) Fragrance 0 3 (mass%)

(2) Wolves extract (extraction method 1, bark) 3.0

(3) Sodium bicarbonate 5 0. 0

(4) Sodium sulfate 4 6. 7 Production method: Mix (1) to (4) uniformly.

[0075] [Example 13] Hair wax

[Table M]

(1) Stearic acid 3.0 (mass%)

(2) Microcrystalline wax 2.0

(3) Cetizoleanolo Cornoret 3.0

(4) Highly polymerized methylpolysiloxane 2.0

(5) Methylpolysiloxane 5.0

(6) Poly (oxyethylene 'oxypropylene) methyl poly! : ^2- xan copolymer

Ten

(7) Methyl paraoxybenzoate 0.1

(8) 1,3—Butyleneglycol 7.5

(9) Arginine 0.7

(10) The balance of purified water 1 0 0

(11) Wolves extract (extraction method 2, bark) 4.0

(12) Fragrance 0. 1 Production method: The oil phase components (1) to (6) are mixed and heated and dissolved at 75 ° C. On the other hand, the aqueous phase components (7) to (10) are dissolved by heating at 75 ° C, and the oil phase component is added and emulsified with a homomixer. Start cooling after emulsification, add ingredients (11) and (12) at 40 ° C and mix evenly

[Example 14] Hair art nick

[Table N]) Ethanol (mass%) Purified water and the remaining wolves extract (extraction method, leaves)

Fragrance Manufacturing method: Mix and homogenize components (1) to (4).

[0077] [Example 15] Tablet

[Table 0]) Corn starch (mass%)) Crystalline cellulose

Carboxymethylcellulose calcium

Caustic anhydride

Magnesium stearate

Wolves's extract (extraction method, leaves) Manufacturing method: (1) to (6) are mixed evenly and compression-molded with a tableting machine to obtain 1 tablet 200mg

[Example 16] Powder

[Table P] Magnesium aluminate (mass%) carboxymethylcellulose calcium

Wolves extract (extraction method, leaves) Production method: After mixing the powders of (1) to (3), they are pulverized by a pulverizer and uniformly dispersed.

[0079] [Example 17] Candy

[Table Q]) Sucrose (mass ⁰ /.)) Minamata

Wolfberry extract (extraction method, leaves)

) Fragrance Manufacturing method: (1) and (2) are heated, mixed, homogenized and cooled, then components (3) and (4) are added at 70 ° C, mixed and homogenized, and then molded.

[0080] [Example 18] Drink agent

[Table R]

(1) Aminoethylsulfonic acid 1 00 0 0 mg

(2) Thiamine nitrate 10 mg

(3) Riboflavin sodium phosphate 5 mg

(4) Pyridoxine hydrochloride 10 mg

(5) Anhydrous caffeine 50 mg

(6) Chenic acid 2 50 mg

(7) D-sorbitol solution 8 g

(8) Wolves extract (extraction method 2, leaves) 10 mg

(9) Fragrance

(10) Amount to make the whole purified water 100 mL Manufacturing method: Add (1) to (9) to (10) sequentially and homogenize.

[0081] According to the present invention, an anti-aging agent, a whitening agent, an antioxidant, and an anti-inflammatory agent having excellent effects can be provided by using ono, mabo or an extract thereof as an active ingredient. wear.

Furthermore, by blending ono, mabo, or its extract into skin preparations (or compositions for external use) such as cosmetics and topical pharmaceuticals, and oral compositions such as foods, it is possible to remove wrinkles, tarmi and skin. It is possible to provide various compositions that exhibit excellent effects in preventing the appearance of various skin symptoms such as skin elasticity, skin weakness, spots and dullness.

[0082] The disclosure of the present application is related to the subject matter described in Japanese Patent Application No. 2006-241623 filed on September 6, 2006, the entire contents of which are incorporated herein by reference.

It should be noted that various modifications and changes may be made to the above-described embodiments without departing from the novel and advantageous features of the present invention other than those already described. Accordingly, all such modifications and changes are intended to be included within the scope of the appended claims.

Claims

The scope of the claims

[1] An anti-aging agent comprising, as an active ingredient, wolfberry (Glias nueberthii) or an extract thereof.

[2] A whitening agent containing Glias nueberthii or an extract thereof as an active ingredient.

[3] Antioxidant containing, as an active ingredient, wolfberry (Glias nueberthii) or its extract.

[4] An anti-inflammatory agent containing Glias nueberthii or an extract thereof as an active ingredient.

[5] A composition comprising a wolfberry (Glias nueberthii) or an extract thereof.

6. The composition according to claim 5, wherein the composition is an external composition or an oral composition.

[7] The composition according to [5] or [6], which is a skin external preparation for improving aging prevention.