JP2003081850A - Skin cosmetic - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an antiinflammatory agent, an antioxidant, an antiaging agent, a skin cosmetic and a bath medicine by discovering a material having antiinflammatory activities, antioxidant activities or antiaging activities from a natural product, and utilizing the material. SOLUTION: The antiinflammatory agent, the antioxidant and the antiaging agent contain an extract of Quassia amara L. as an active ingredient. The skin cosmetic and the bath medicine is obtained by formulating the extract.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an anti-inflammatory agent, an antioxidant and an anti-aging agent containing a plant extract as an active ingredient, and a skin cosmetic and a bath preparation containing the plant extract.

[0002]

2. Description of the Related Art The causes and onset mechanisms of inflammatory diseases such as contact dermatitis (rash), psoriasis, pemphigus vulgaris, and various other skin diseases associated with rough skin are diverse, and the causes are various. Platelet aggregation and histamine release by click AMP phosphodiesterase are known. In addition, by suppressing the action of cyclic AMP phosphodiesterase, which is an enzyme that decomposes cyclic AMP that is involved in fat metabolism promotion, the intracellular cyclic AMP concentration rises, fat metabolism becomes active, and obesity increases. Is known to be resolved.

Platelet aggregation causes activation of phospholipase A2 of the arachidonic acid cascade, and leukotriene B4, prostaglandin E2, etc. released by the activation cause inflammatory reaction. Therefore, substances that inhibit or suppress platelet aggregation prevent or prevent allergic or inflammatory diseases.
Attempts have been made to treat it, and aspirin, ticlopidine, sulfipyrazone, etc. have been used as such platelet aggregation inhibitors. However, all of these substances have side effects, which has been a problem in terms of safety.

[0004] Further, platelet aggregation is related to the concentration of cyclic AMP in platelets, and when cyclic AMP is decomposed by cyclic AMP phosphodiesterase and the concentration of cyclic AMP decreases, platelets easily aggregate. Therefore, it is considered that platelet aggregation can be prevented by suppressing the action of cyclic AMP phosphodiesterase to prevent the decrease of the cyclic AMP concentration.

On the other hand, in recent years, active oxygen has been attracting attention as a factor that particularly oxidizes biological components, and its adverse effect on the living body has become a problem. Active oxygen is generated in the process of energy metabolism in living cells, and includes superoxide (that is, superoxide anion (.O _2- ) generated by one-electron reduction of oxygen molecule, hydrogen peroxide (H ₂ O ₂ ), singlet). There are term oxygen ( ¹ O ₂ ) and hydroxy radicals (.OH).
It is known that singlet oxygen is particularly reactive among active oxygen and directly produces lipid peroxide related to inflammation and aging, and elastin, which plays an important role in maintaining skin firmness and elasticity, By causing cross-linking and fragmentation of collagen, it reduces the elasticity of the skin and causes wrinkles and tarmi.

As the cause of aging such as wrinkle formation of the skin and reduction of elasticity of the skin, various causes other than active oxygen are considered. For example, hyaluronic acid is a type of acidic mucopolysaccharide in which N-acetylglucosamine and glucuronic acid are alternately bound, and has a function of retaining water in the interstitial spaces of the skin, and removes water from subcutaneous connective tissue. It is considered that when the abundant amount of hyaluronic acid is decreased, the freshness of the skin is lost and wrinkles are formed. Therefore,
It is thought that by promoting hyaluronic acid synthesis in dermal fibroblasts, it is possible to increase the water retention capacity of the skin and, as a result, to make the skin firm, glossy and elastic.

One of the causes of skin aging with aging is that the secretion of estrogen, which is a female hormone, is decreased. That is, estrogen is deeply involved in maintaining the health of adult women, and its insufficient secretion leads to various medical disorders, skin hypersensitivity, decreased elasticity, decreased moisture, etc.
It is known to cause unwanted skin changes.

[0008]

[Problems to be Solved by the Invention] In order to inhibit / suppress an inflammatory reaction and prevent / treat an inflammatory disease, inhibition / suppression of cyclic AMP degradation caused by cyclic AMP phosphodiesterase, histamine release, etc., which is the cause of the inhibition, is suppressed. To be useful.

In addition, by inhibiting or suppressing the generation of active oxygen (particularly singlet oxygen), it is possible to prevent or treat skin aging such as wrinkle formation or elasticity reduction by suppressing the production of lipid peroxide. it is conceivable that.

Furthermore, by promoting hyaluronic acid synthesis in dermal fibroblasts and compensating for decreased estrogen secretion, it is possible to prevent / treat skin aging such as wrinkle formation and elasticity reduction. Conceivable.

Therefore, the present invention firstly finds, from natural products, those having a cyclic AMP phosphodiesterase inhibitory action and / or histamine release inhibitory action, and provides an anti-inflammatory agent containing the same as an active ingredient. With the goal.

A second object of the present invention is to find out a natural product having a singlet oxygen scavenging action, and to provide an antioxidant containing it as an active ingredient.

Thirdly, the present invention is to find a natural product having hyaluronic acid synthesis accelerating action and / or estrogen-like action, and to provide an anti-aging agent containing the same. And

Fourthly, the present invention is to find out among natural products, those having an anti-inflammatory action, an antioxidant action or an anti-aging action, and to provide skin cosmetics and bath agents containing them. To aim.

[0015]

In order to achieve the above-mentioned object, the anti-inflammatory agent, antioxidant or anti-aging agent of the present invention contains an extract of Querus chinensis (Quassia amara L.) as an active ingredient. Is characterized by.

In the anti-inflammatory agent of the present invention, it is preferable that the extract has a cyclic AMP phosphodiesterase inhibitory action and / or a histamine release inhibitory action. Further, in the antioxidant of the present invention, it is preferable that the extract has a singlet oxygen scavenging action. Further, in the anti-aging agent of the present invention, it is preferable that the extract has a hyaluronic acid synthesis promoting action and / or an estrogen-like action.

In order to achieve the above object, the skin cosmetic or bath agent of the present invention comprises
a amara L.) extract.

[0018]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described in detail below. In the present invention, "American lacquer extract" means an extract obtained from a Chinese lacquer extract as a raw material,
Diluents or concentrates of the extract, dried products obtained by drying the extract, or crude or purified products of these are included.

The plant used as the extraction raw material is American oyster (Quassia amara L.). American oyster (Qu
assia amara L.) is a plant belonging to the family Littoridae, a very bitter shrub native to tropical America and is widely cultivated in the tropics for medicinal purposes. Wood and bark are sometimes used as antipyretics and antiparasitics, but their anti-inflammatory, antioxidant or anti-aging effects have not been known.

The constituent parts of the plant used as the extraction raw material are not particularly limited, and for example, constituent parts such as leaves, stems, bark, flowers, roots and fruits can be used as the extraction raw material, and the whole plant can be used. Can be used as an extraction raw material, but it is particularly preferable to use bark as an extraction raw material.

[0021] The details of the anti-inflammatory substance, antioxidant substance or anti-aging substance contained in the extract of Pleurotus cornucopiae are unknown, but the anti-inflammatory action from Pleurotus cornucopiae by an extraction method generally used for plant extraction, An extract having an antioxidant effect or an anti-aging effect can be obtained. For example, it can be obtained by drying the extraction raw material and then pulverizing it as it is or by using a coarse crusher and subjecting it to extraction with an extraction solvent. At this time, the extraction raw material may be dried in the sun or using a commonly used dryer. In addition, the American oyster may be used as an extraction raw material after being subjected to a pretreatment such as degreasing with a non-polar solvent such as hexane and benzene. By performing a pretreatment such as degreasing, the extraction treatment with the polar solvent from the American oyster can be efficiently performed.

As the extraction solvent, it is preferable to use water, a hydrophilic organic solvent or a mixture thereof at room temperature or at a temperature not higher than the boiling point of the solvent.

The water that can be used as the extraction solvent includes pure water, tap water, well water, mineral water, mineral water, hot spring water, spring water, fresh water and the like, as well as those that have been subjected to various treatments.
Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering and the like.
Therefore, the water that can be used as the extraction solvent in the present invention also includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered saline and the like.

Examples of the hydrophilic organic solvent which can be used as the extraction solvent include, for example, methanol, ethanol, propyl alcohol, isopropyl alcohol and the like having 1 to 4 carbon atoms.
And lower aliphatic ketones such as acetone and methyl ethyl ketone; and polyhydric alcohols such as 1,3-butylene glycol, propylene glycol and glycerin.

When a mixture of water and a hydrophilic organic solvent is used as an extraction solvent, water is used in the case of a lower alcohol.
1 to 90 parts by weight with respect to 0 parts by weight, 1 to 40 parts by weight with respect to 10 parts by weight of water in the case of a lower aliphatic ketone, and 10 to 90 parts by weight with respect to 10 parts by weight of water with a polyhydric alcohol. It is preferable to add.

In the present invention, it is not necessary to employ a special extraction method in order to obtain an extract having an anti-inflammatory action, an antioxidant action or an anti-aging action, and extraction is carried out at room temperature or under reflux heating using any device. be able to.

Specifically, the extraction raw material is put into a treatment tank filled with an extraction solvent, the soluble components are eluted with occasional stirring if necessary, and then the solid matter is removed by filtration to obtain the obtained extraction. The extract is obtained by distilling off the extraction solvent from the liquid and drying. The amount of extraction solvent is usually 5 to 1 of the extraction raw material.
The amount is 5 times (mass ratio), and the extraction temperature is usually from room temperature to 95.
C., and the extraction time is usually about 1 to 24 hours.

The obtained extract is diluted, concentrated, dried according to a conventional method in order to obtain a diluted solution or concentrated solution of the extracted solution, a dried product of the extracted solution, or a crudely purified product or a purified product thereof. You may perform processing, such as refinement | purification.

The obtained extract is an anti-inflammatory agent,
It can be used as an antioxidant or an anti-aging agent, but a concentrated solution or a dried product thereof is easier to use. The American oyster extract can be formulated according to a conventional method. In the case of formulation, a pharmaceutically acceptable carrier such as dextrin or cyclodextrin and any other auxiliaries can be added to facilitate storage and handling. It can be formulated into any dosage form such as a tablet, tablet and the like.

Further, since the extract of Pleurotus cornucopiae has a peculiar odor, it is possible to carry out purification for the purpose of decolorization, deodorization, etc. within a range that does not impair its physiological activity. Since it is not used in a large amount when it is added to a raw material, there is no practical problem even if it is left unpurified. Specifically, the purification can be performed by activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, liquid-liquid countercurrent distribution, or the like.

The extract of Pleurotus cornucopiae has an anti-inflammatory action, an antioxidant action or an anti-aging action and can be used as an anti-inflammatory agent, an antioxidant or an anti-aging agent by utilizing each action. .

Here, the anti-inflammatory action of the extract of Pleurotus cornucopiae is exerted based on the cyclic AMP phosphodiesterase inhibitory action and / or histamine release inhibitory action. Since the extract of Pleurotus cornucopiae has a cyclic AMP phosphodiesterase inhibitory activity and / or a histamine release inhibitory activity, it can also be used as an active ingredient of a cyclic AMP phosphodiesterase inhibitor or a histamine release inhibitor. An AMP phosphodiesterase inhibitor and / or a histamine release inhibitor may be used as the active ingredient of the anti-inflammatory agent of the present invention.

The antioxidative action of the extract of Pleurotus cornucopiae is exerted based on the singlet oxygen scavenging action. The extract of Pleurotus cornucopiae exhibits a scavenging action specifically to singlet oxygen among active oxygen. Since the extract of Pleurotus cornucopiae has a singlet oxygen scavenging action, it can be used as an active ingredient of a singlet oxygen scavenger, and this singlet oxygen scavenger is used as an active ingredient of the antioxidant of the present invention. Good.

The anti-aging action of the extract of Pleurotus cornucopiae is exerted based on the hyaluronic acid synthesis promoting action and / or the estrogenic action. In other words, Aedes aegypti extract is percutaneously absorbed, reaches fibroblasts, promotes the synthesis of hyaluronic acid, and causes the dermal layer to be supplemented with sufficient hyaluronic acid. It has the effect of preventing and improving the aging of the skin due to the deterioration of skin. Since the extract of Pleurotus cornucopiae has a hyaluronic acid synthesis accelerating action and / or an estrogen-like action, it can also be used as an active ingredient of a hyaluronic acid synthesis accelerating agent or an estrogen-like acting agent. And / or an estrogenic agent may be an active ingredient of the anti-aging agent of the present invention.

The Chinese lacquer extract is useful for preventing / ameliorating inflammatory diseases and preventing / ameliorating skin aging.
It has excellent usability and safety when applied to the skin, and since it is well absorbed transdermally, it is suitable for incorporation into skin cosmetics or bath agents. Any one of the anti-inflammatory agent, antioxidant or anti-aging agent of the present invention may be blended in the skin cosmetic or bath agent, or two or more thereof may be blended in combination.

There are no particular restrictions on the skin cosmetics which can be blended with the extract of Pleurotus cornucopiae, and specific examples thereof include ointments, creams, emulsions, lotions, packs, lipsticks, bath agents and the like.

The compounding amount of the American oyster extract in the skin cosmetics or bath agents can be appropriately adjusted depending on the kind of the skin cosmetics, the physiological activity of the extract, etc., but a suitable compounding ratio is a standard extract. Converted to about 0.001-1
It is 0% by weight, preferably about 0.05 to 2% by weight.

The skin cosmetics or bath preparations of the present invention are generally used in the production of skin cosmetics or bath preparations as long as they do not interfere with the anti-inflammatory action, anti-oxidant action and anti-aging action of the extract of Pleurotus cornucopiae. Various main agents and auxiliaries, and other optional auxiliaries can be used. The skin cosmetics or bath preparations of the present invention are not limited to those in which the extract of Pleurotus cornucopiae is the only main ingredient for prevention / treatment of inflammatory diseases and prevention / improvement of skin aging.

Specific examples of those that can be used as a skin cosmetic composition component or a bath agent composition component together with the American bittern extract include avocado oil, rice bran oil, rice germ oil,
Oily ingredients such as lanolin and squalane; glycerin, 1,
Moisturizers such as 3-butylene glycol, collagen, hyaluronic acid and salts thereof, chondroitin sulfate and salts thereof, chitosan and chitin; complex lipids such as glycerophospholipids, sphingolipids, glyceroglycolipids and glycosphingolipids; SOD, catalase, β -Carotene, Ginkgo biloba extract, Vitamin C and its derivatives, Scutellaria extract,
Active oxygen scavenging substances such as kujin extract; guaiazulene, kamaazulene and its derivatives; anti-inflammatory agents such as glycyrrhizinic acid, glycyrrhetinic acid and its salts, glycyrrhetinic acid derivatives, zinc oxide; other various plant extracts, thickeners, preservatives Agents, ultraviolet absorbers, fragrances, antioxidants, water, alcohols and the like.

The anti-inflammatory agent, anti-oxidant, anti-aging agent, skin cosmetic and bath agent of the present invention described above are preferably applied to humans, but they exhibit their respective effects. So long as it is applicable to non-human animals.

[0041]

EXAMPLES The present invention will be specifically described below with reference to production examples, test examples and formulation examples, but the present invention is not limited to the following examples.

[Manufacturing Example 1] 10 g of dried crushed stem parts of the Japanese bitter oyster were added to 50 g of water as an extraction solvent, respectively.
% Ethanol (weight ratio of water to ethanol 1: 1) and 100 mL of ethanol were added, and reflux extraction was performed at 80 ° C. for 2 hours. After that, the extract obtained by filtration is concentrated under reduced pressure,
Further, it was dried with a vacuum dryer to obtain a powdery extract. The yield of the extract was as shown in Table 1.

[Table 1] Sample No. Extraction Solvent extraction rate (%) 1 Water 10.1 2 50% Ethanol 15.0 3 Ethanol 11.2

[Test Example 1] Hyaluronic acid synthesis promoting action test
Examination Normal human neonatal fibroblast (NB1RGB) 1 x 10⁶The individual
75 cm^TwoΑ-M containing 10% FBS using a flask
EM medium (GIBCO) (pH 7.2) at 37 ° C, 5% C
O_TwoIt was cultured for 7 days under the condition of -95% air. Tripe
Cells were collected by syn-treatment and α-ME containing 1% FBS.
Cell concentration is 2.2 x 10 using M medium ^FourAdjust to pieces / mL
Prepare and seed 100 μL each on a 96-well microplate.
37 ° C, 5% CO_TwoCultivated overnight under -95% air condition
I nourished.

On the next day, 100 ml of α-MEM medium containing 1% FBS containing the sample (200 μg / mL) was dissolved in each well.
Each μL was added, and the cells were cultured for 3 days under the conditions of 37 ° C., 5% CO ₂ -95% air. Α-without sample as control
The MEM medium was added to each well in an amount of 100 μL. Quantification of hyaluronic acid produced by ELISA
The method was used as follows. That is, 10 μL of this culture supernatant was diluted 10-fold with 90 μL of PBS (−),
50 μL thereof was added to an ELISA plate coated with hyaluronic acid in advance, and ELISA was performed using various antibodies (anti-keratan sulfate antibody (Seikagaku Corporation), peroxidase-labeled anti-mouse IgG antibody) to obtain a calibration curve. Was used to quantify hyaluronic acid. The hyaluronic acid synthesis rate in the control was calculated as 100%, and the hyaluronic acid synthesis rate (%) when each sample was added was calculated. The results are shown in Table 2.

[0046] [Table 2] Sample NO. Extraction was hyaluronic acid synthesis rate (%) 1 water extract 147.5 2 50% ethanol extract 166.8 3 ethanol extract 135.4

From the results shown in Table 2, it was confirmed that the extract of Pleurotus cornucopiae has a hyaluronic acid synthesis promoting action.

[Test Example 2] Singlet oxygen scavenging test 2% red blood cell suspension 5 in a transparent glass bottle (10 mL volume)
mL, 5 mL of a pH 7.4 isotonic phosphate buffer containing a sample at a predetermined concentration, and 0.01 mL of a photosensitizer (10 mM hematoporphyrin-20 mM sodium hydroxide solution) were mixed. The resulting solution was placed on a merry-go-round for 7.5
It was uniformly irradiated with a W halogen lamp for 35 minutes to generate singlet oxygen ( ¹ O ₂ ), which caused hemolysis of red blood cells. 1 mL of this reaction solution was collected, 2 mL of isotonic phosphate buffer was added and mixed, and then centrifuged at 3000 rpm for 5 minutes at 4 ° C. Then, the supernatant was collected and the absorbance at a wavelength of 540 nm was measured.

Separately, 1 mL of the above reaction solution in which red blood cells were partially hemolyzed was taken, and 2 mL of distilled water was added thereto to completely hemolyze, and the absorbance was similarly measured. From the measured absorbance, the singlet oxygen scavenging rate was calculated by the following formula.

[0050] Singlet oxygen scavenging rate (%) = (1−B / A) × 100

In the above formula, "A" represents the absorbance of the control and "B" represents the absorbance of the reaction solution supernatant.

The scavenging rate was measured by gradually reducing the sample concentration, and the sample concentration (ppm; μg / mL) at which the scavenging rate of singlet oxygen was 50% was determined by the interpolation method. The results are shown in Table 3.

[0053] [Table 3] Sample NO. Extraction was 50% inhibition sample concentration (ppm) 1 water extract 234.1 2 50% ethanol extract 200.0 3 ethanol extract 228.6

From the results shown in Table 3, it was confirmed that the extract of Pleurotus cornucopiae has a singlet oxygen scavenging action. It was also confirmed that the degree of this singlet oxygen scavenging action can be adjusted by the concentration of the extract.

[Test Example 3] Estrogen-like action test Th for examining the effect on proliferation of estrogen-dependent cells Th
omas et al. (In Vitro Cell. Dev. Biol. 28A, 595-602,
1992). M derived from human breast cancer
CF-7 cells were cultured in a 75 cm ² flask until they became confluent and treated with trypsin to give this MCF.
-7 cells were collected, 10% FBS (treated with activated carbon), 1
MEM medium containing% NEAA and 1 mM sodium pyruvate but not phenol red (hereinafter, "MEM
Abbreviated as "medium"), 3 × 10 ⁴ cells / mL
Was prepared.

The prepared MCF-7 cells were seeded in a 24-well plate in an amount of 0.9 mL each, and 37
The cells were cultured at 5 ° C and 5% CO2-95% air. After 6 hours (day 0), 10 times the final concentration (1
100 μL of the sample solution adjusted to 25 ppm) was added to the plate, and the culture was continued.

Six days after the start of culture, the medium was 0.97 mm
The medium was replaced with a MEM medium containing ol / L MTT, and after culturing for 2 hours, the medium was replaced with isopropanol to extract blue formazan produced in the cells. Regarding the eluted isopropanol containing blue formazan, the absorbance at 570 nm at which the absorption maximum of blue formazan is present was measured. In addition, in order to correct the effect of adherent cells, 6
The absorbance at 50 nm was also measured, and the difference between the two absorbances was used as a value proportional to the amount of blue formazan produced (the absorbance in the following formula is the corrected absorbance).

As a positive control, 0.02 ppm ethinyl estradiol was used. The strength of the estrogen-like action (estrogen-dependent proliferative action) was calculated by the following equation with the absorbance at 100% when no sample was added. The results of this test are shown in Table 4.

[0059] Estrogen-like action (%) = A / B x 100

In the above formula, "A" represents the absorbance when the sample was added, and "B" represents the absorbance when the sample was not added.

[0061] [Table 4] Sample NO. Extraction was estrogenic rate (%) 1 water extract 108.2 2 50% ethanol extract 119.5 3 ethanol extract 112.1

From the results shown in Table 4, it was confirmed that the extract of Pleurotus cornucopiae has an estrogen-like action.

Test Example 4 Histamine Release Inhibitory Test The histamine release inhibiting action was tested by the following test method. In addition, the following test method is a test method that evaluates histamine release inhibitory activity using hexosaminidase release as an index, by utilizing the fact that intracellular histamine is released and hexosaminidase is released at the same time. is there.

25 cm^TwoMedium in culture flask
RB in (S-MEM medium containing 15% FBS; the same applies below)
L-2H3 cell 1.0 × 10⁶Seed, 37 ° C, 5
% CO _TwoCultured under -95% air for 4 days. Then
Treat with trypsin and centrifuge (800 rpm, 4 minutes)
Then the cells were collected. The cells obtained were 4.0 × 10⁵ce
It is suspended in the medium at 11 / mL and the mouse monoclonal
Anti-dinitrophenyl group IgE (DNP-Specif
ic IgE) was added at a concentration of 0.5 μg / mL.
Add 100μ of this cell suspension to each well of a 96-well plate.
L seeds at 37 ° C, 5% CO_Two-95% below air
The cells were cultured for 24 hours. After completion of culture, remove the medium in each hole
And washed twice with Silaghanian buffer. Next, the above buffer
Add 30 μL of the solution and 10 μL of the sample solution, and add 1 at 37 ° C.
Incubated for 0 minutes. Then dinitrophenyl
Add 10 μL of modified bovine serum albumin (DNP-BSA)
And further incubated at 37 ° C for 15 minutes. So
After that, add 10 μL of the supernatant to a new 96-well plate under ice cooling.
Transferred to this, 1 mmol / L p-nitrophenyl
10 μL of -N-acetyl-β-D-glucosamide solution
In addition, it incubated at 37 degreeC for 1 hour. End of reaction
After completion, 0.1 mol / L Na_TwoCO_Three-Na _TwoHCO
_ThreeAdd 250 μL of solution and place in a microplate reader.
Absorbance measurement at 415 nm against 650 nm
The measurement was performed (the absorbance measured at this time is A).

Further, the same treatment and absorbance measurement were performed on the cell supernatant to which the Silaghanian buffer was added instead of the sample solution (the absorbance measured at this time is B).

In addition, the cell supernatant and 0.1 mol / L Na
On ₂ CO ₃ -Na ₂ HCO ₃ solution obtained by reacting the same processing is also performed absorbance measurement (absorbance measured this time is C).

Further, the same processing and absorbance measurement were carried out by DNP-
The experiment was also carried out with the addition of a Silaghanian buffer solution instead of BSA (the absorbance measured at this time is defined as D). And the hexosaminidase release inhibition rate (%) according to the following formula
I asked.

Release inhibition rate (%) = [1-{(A-C-
D) / (BD)}] × 100

The above release inhibition rate was measured by gradually decreasing the sample concentration, and the release of hexosaminidase was 50%.
The inhibitory sample concentration (ppm; μg / mL) was determined by interpolation. The results are shown in Table 5.

[0070] [Table 5] Sample NO. Extraction was 50% inhibition sample concentration (ppm) 1 water extract 350 2 50% ethanol extract 316 3 ethanol extract 375

From the results shown in Table 5, it was confirmed that the extract of Pleurotus cornucopiae had a histamine release inhibitory action. It was also confirmed that the degree of histamine release inhibitory effect can be adjusted by the concentration of the extract.

Test Example 5 Cyclic AMP Phosphodiesterase Inhibition Test 0.2 mL of Tris-HCl buffer (pH 7.5) containing 5 mM magnesium chloride was added to 0.1 mL of fetal serum arpmin solution and 0.1 mL of cyclic AMP phosphodiesterase solution. Is added, and the sample solution is 0.05 m
L was added and preincubated at 37 ° C. for 5 minutes. Then, 0.05 mL of cyclic AMP solution was added, and the mixture was incubated at 37 ° C. for 60 minutes. Boil for 3 minutes in a boiling bath to stop the reaction, then at 4 ° C for 3500 r
The reaction substrate 5'-AMP in the supernatant was quantified by high performance liquid chromatography. The same enzymatic reaction and reaction substrate analysis are performed without adding the sample solution, and the cyclic AMP phosphodiesterase inhibition rate (%) of the sample is determined from the ratio of the reaction group mass when the sample is added to the reaction group mass when the sample is not added. I asked.

The above-mentioned measurement was repeated by gradually decreasing the sample concentration of the sample solution, and the sample concentration IC at which the activity of cyclic AMP phosphodiesterase was inhibited by 50%
₅₀ (ppm; μg / mL) was determined by the interpolation method. The results are shown in Table 6.

[0074] [Table 6] Sample NO. Extraction was 50% inhibition sample concentration (ppm) 1 water extract 50.0 2 50% ethanol extract 45.0 3 ethanol extract 58.5

From the results shown in Table 6, it was confirmed that the extract of Pleurotus cornucopiae had a cyclic AMP phosphodiesterase inhibitory action. Also, this cyclic A
It was confirmed that the degree of MP phosphodiesterase inhibitory effect can be adjusted by the concentration of the extract.

[Test Example 6] An emulsion containing the 50% ethanol extract obtained in Production Example 1 (hereinafter referred to as "Example emulsion") was prepared according to a conventional method. The composition of the example emulsion is shown below.

[0077]     American oyster 50% ethanol extract 0.5g     Cetyl alcohol 0.5g     2.0 g of beeswax     Polyoxyethylene sorbitan oleate (20E.0) 1.0g     Glycerin monostearate 1.0 g     Sodium hyaluronate 0.1g     Propylene glycol 5.0g     Ethanol 3.0g     Ethyl paraben 0.3g     Fragrance 0.03g     Purified water balance (total volume is 100 ml)

The following evaluation tests were carried out on the emulsion of the example and the emulsion of the comparative example having the same composition as the emulsion of the example except that it did not contain the extract of the oyster extract. Subjects consisting of 20 women with rough skin (ages 27 to 52 years old) were selected. For each subject, the example emulsion was applied to the left half of the face, the comparative emulsion was applied to the right half, and three times each morning and evening. It was applied to the facial cheeks for a month. Table 7 shows the results of making each subject give their impressions about the skin condition after 3 months of use.

[Table 7] Evaluation item Example Example Comparative emulsion Comparative emulsion No superiority or inferiority Moisture appeared on the skin 20 0 0 Skin stains and dullness disappeared 15 2 3 Skin wrinkles improved 14 people 1 person 5 people

As shown in Table 7, in the area coated with the emulsion of Example, the skin roughness (skin aging) was remarkably improved as compared with the area coated with the emulsion of Comparative Example. From this, it was confirmed that the extract of Pleurotus cornucopiae has an anti-aging effect on the skin.

[Formulation Example 1] A lotion having the following composition was produced by a conventional method. American oyster ethanol extract (Production Example 1) 0.5 g Glycerin 4.0 g 1,3-Butylene glycol 4.0 g Ethanol 7.0 g Polyoxyethylenesorbitan oleate (20E.0) 0.5 g Methylparaben 0.05 g Citric acid 0.01 g Sodium citrate 0.1 g Perfume 0.05 g Purified water balance (total volume is 100 ml)

[Formulation Example 2] A cream having the following composition was produced by a conventional method. Water extract of Pleurotus cornucopiae (Production Example 1) 1.5 g Cetostearyl alcohol 4.0 g Squalane 40.0 g Beeswax 3.0 g Reduced lanolin 5.0 g Ethylparaben 0.3 g Polyoxyethylene sorbitan oleate (20E.O) 2. 0 g Glycerin monostearate 2.0 g 1,3-butylene glycol 5.0 g Perfume 0.03 g Glycerin 5.0 g Purified water balance (total amount is 100 ml)

[Formulation Example 3] A pack having the following composition was produced by a conventional method. American oyster 50% ethanol extract (Production Example 1) 6.0 g Polyvinyl alcohol 15.0 g Polyethylene glycol 3.0 g Propylene glycol 7.0 g Ethanol 10.0 g Methylparaben 0.05 g Perfume 0.05 g Purified water balance (total volume 100 ml and Do)

[Formulation Example 4] A bath agent having the following composition was produced by a conventional method. American oyster ethanol extract (Production Example 1) 2.0 g Sorbitan monolaurate 1.0 g Polyoxyethylene sorbitan oleate (20E.O) 1.5 g Carboxyvinyl polymer 0.05 g Methylparaben 0.1 g 1,3-butylene glycol 10 0.0 g denatured alcohol 10.0 g Fragrance small amount purified water balance (total amount is 100 ml)

[0085]

INDUSTRIAL APPLICABILITY According to the present invention, an anti-inflammatory agent, an antioxidant or an anti-aging agent is provided. Further, according to the present invention, a skin cosmetic and a bath preparation having an anti-inflammatory action, an antioxidant action or an anti-aging action are provided. The anti-inflammatory agent of the present invention can effectively prevent or improve inflammation associated with cyclic AMP degradation by cyclic AMP phosphodiesterase, histamine release, generation of singlet oxygen, and the like. Further, according to the antioxidant of the present invention, it is possible to effectively prevent and treat aging phenomena such as formation of wrinkles of the skin and reduction of elasticity through the prevention of oxidation of biological components due to the singlet oxygen scavenging action. . Further, according to the anti-aging agent of the present invention, it is possible to effectively prevent / treat aging phenomena such as formation of wrinkles of the skin and decrease in elasticity through the hyaluronic acid synthesis promoting action and / or the estrogenic action. The anti-inflammatory agent, antioxidant and anti-aging agent of the present invention have excellent usability and safety when applied to the skin, and therefore are suitable for incorporation in skin cosmetics or bath agents. The skin cosmetics and bath preparations of the invention are useful for preventing and / or improving skin inflammation and aging.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ identification code FI theme code (reference) A61K 7/48 A61K 7/48 7/50 7/50 A61P 5/30 A61P 5/30 29/00 29 / 00 39/06 39/06 43/00 43/00 111 111 113 113 113 (72) Inventor Naoko Kishida 14703-10 Mutocho, Onomichi, Hiroshima Prefecture Maruzen Pharmaceutical Co., Ltd. (72) Nobuaki Oto For Onomichi, Hiroshima Prefecture Higashimachi 14703-10 Maruzen Pharmaceutical Co., Ltd. F-term (Reference) 4C083 AA082 AA111 AA112 AC022 AC072 AC102 AC122 AC302 AC422 AC442 AC482 AD042 AD092 AD112 AD512 CC02 CC04 CC05 CC07 CC25 DD23 DD27 DD31 4C088 AB12 AC01 AC03 AC04 AC05 AC06 AC11 CA03 MA63 MA89 ZB11 ZC13 ZC20 ZC52

Claims (8)

[Claims] 1. An anti-inflammatory agent, which comprises an extract of American oyster (Quassia amara L.) as an active ingredient. 2. The anti-inflammatory agent according to claim 1, wherein the extract has a cyclic AMP phosphodiesterase inhibitory action and / or a histamine release inhibitory action. 3. An antioxidant, which contains an extract of Quercus chinensis (Quassia amara L.) as an active ingredient. 4. The antioxidant according to claim 3, wherein the extract has a singlet oxygen scavenging action. 5. An anti-aging agent, which comprises an extract of Quercus chinensis (Quassia amara L.) as an active ingredient. 6. The anti-aging agent according to claim 5, wherein the extract has a hyaluronic acid synthesis promoting action and / or an estrogen-like action. 7. A skin cosmetic composition comprising an extract of American oyster (Quassia amara L.). 8. A bath agent comprising an extract of American oyster (Quassia amara L.).