JP2010168400A - Antiphlogistic and skin cosmetic - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a multi-purpose antiphlogistic whose effective components are highly safe substances derived from naturally occurring materials. <P>SOLUTION: The antiphlogistic includes an extract from the bark of Schima wallichii or the bark of Quercus semicarpifolia and not less than two kinds selected from among phospholipase A2 inhibitors, hyaluronidase inhibitors, and histamine release inhibitors as effective components. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to an anti-inflammatory agent useful for preventing skin inflammation due to various causes and used in a skin cosmetic composition, and a skin cosmetic composition.

  There are various causes and mechanisms of contact dermatitis (rash), psoriasis, pemphigus vulgaris, congenital chicken pox, and various other skin diseases with rough skin, but the main ones and countermeasures are as follows: There was something.

(1) By phospholipase A2 Phospholipase A2 is an important enzyme of the arachidonic acid cascade, which is the metabolic pathway of arachidonic acid. When the action of this enzyme is activated, abnormalities in arachidonic acid metabolism occur, causing inflammation, allergy, asthma, false Causes blood, myocardial infarction, etc. Therefore, attempts have been made to deal with the above disorders with substances that inhibit this enzyme, and mepacrine, parabromophenacyl bromide, dexamethasone, and the like have been used. However, these are synthetic products, and side effects have been a problem.

(2) By hyaluronidase Hyaluronidase is an enzyme that hydrolyzes hyaluronic acid, but hyaluronic acid is a mucopolysaccharide found in mesenchymal tissues, and it helps prevent the invasion and transmission of microorganisms and poisons. When the action of hyaluronidase is activated, damage due to the decrease in the protective action by hyaluronic acid is likely to occur. It is also related to type I allergic reaction and is considered to be an enzyme that governs degranulation from mast cell. Thus, anti-inflammatory agents and antiallergic agents using substances that inhibit the action of hyaluronidase, such as tranilast, DSCG, baicalein and the like, have been provided, but side effects have also been a problem.

(3) Due to histamine release Histamine is present in mast cells, and the substance released by degranulation reaction acts as an inflammatory substance. Examples of substances effective for suppressing allergic symptoms by inhibiting the release of histamine include tranilast, sodium cromoglycate, baicalin, baicalein, promethazine hydrochloride and the like. However, these substances also have side effects, and there has been a problem in terms of safety when blended into general skin cosmetics.

  Furthermore, since the conventional drugs effective as anti-inflammatory agents were effective only for any one of phospholipase A2 inhibition, hyaluronidase inhibition, histamine release suppression, etc., the anti-inflammatory agent is effective against unspecified inflammation. In order to obtain inflammatory agents and skin cosmetics, it was necessary to use a plurality of drugs in combination.

  An object of the present invention is to find a substance that is effective against many types of inflammation from among highly safe substances derived from natural products, and to provide it as a multipurpose anti-inflammatory agent.

  Another object of the present invention is to provide an anti-inflammatory skin cosmetic comprising a plurality of substances effective against the onset of inflammation.

  The anti-inflammatory agent according to the present invention is extracted from the bark of Mobokuju or the bark of Kouzalenki and contains any two or more of a phospholipase A2 inhibitor, a hyaluronidase inhibitor, and a histamine release inhibitor as an active ingredient. .

  The skin cosmetic according to the present invention is obtained by blending the anti-inflammatory agent according to the present invention.

  As described above, according to the present invention, there are provided anti-inflammatory agents that are effective in preventing and improving various skin diseases, rough skin and the like that involve inflammatory enzymes, such as phospholipase A2 and hyaluronidase, and are excellent in safety. be able to.

  First, a method for obtaining an anti-inflammatory substance used as an active ingredient of the anti-inflammatory agent according to the present invention will be described. Plants that can be extracted from anti-inflammatory substances are as follows.

(1) Hydrangea (scientific name: Podocarpus rifolios )
It is an evergreen Takagi from the family Maki. As wood, it is used to make furniture and ships. It is the stems and leaves of the anti-inflammatory substance that can be extracted.

(2) Koukatsutsuji (scientific name: Rhododendron arborum )
An azalea shrub. The flower and bark are used as the extraction raw material for the anti-inflammatory substance. It is said that the dried flowers are effective in treating bronchitis, irregular menstrual flow, and digestive tract bleeding. To some extent bark is used for the core of the lamp.

(3) Moubokuju (scientific name: Schima wallichii)
It is an evergreen small Takagi of the camelliaceae. The bark portion is used as an extraction raw material for the anti-inflammatory substance, and it is known that this bark can be used as a fish poison.

(4) Sarasouju (scientific name: Shorea robusta )
It is a deciduous Takagi of the Dipterocarpaceae family. Flowers are drunk as tea, and wood is used as building materials and railway sleepers. The bark portion is used as a raw material for extracting the anti-inflammatory substance.

(5) Kozanreki (Scientific name: Quercus semicarpifolia )
It is an evergreen or semi-evergreen green tree of the beech family. Although leaves and seeds are said to have a detoxifying action and are effective in treating enteritis and the like, it has not been known that the bark contains an anti-inflammatory substance.

  The anti-inflammatory substance contained in the extraction raw material is an organic solvent, for example, a lower aliphatic alcohol such as methanol or ethanol (which may be a hydrous alcohol), a polar solvent such as acetone, 1,3-butylene glycol, or propylene glycol. In addition, it can be extracted at room temperature or under heating using an intermediate polar solvent such as chloroform or ethyl acetate.

It has been confirmed that the extracts obtained from the respective extraction raw materials have a plurality of actions effective for anti-inflammation as described below (see Examples below).
Chrysanthemum extract: Phospholipase A2 inhibition, Histamine release inhibition Koutsutsuji extract: Phospholipase A2 inhibition, Hyaluronidase inhibition, Histamine release inhibition Mud extract: Phospholipase A2 inhibition, Hyaluronidase inhibition Salamander extract: Hyaluronidase inhibition Kozanreze extract inhibition A2 inhibition, hyaluronidase inhibition, histamine release inhibition

  Since these active ingredients are substances that are easily absorbed percutaneously, they can be used for blending appropriate amounts in skin cosmetics and the like to reach the inflamed site from the skin surface and to calm the inflammation.

  Specific examples of skin cosmetics suitable for incorporating the anti-inflammatory agent according to the present invention include ointments, pops, creams, emulsions, lotions, packs, jelly and the like.

  When the skin cosmetics are produced by blending the anti-inflammatory agent of the present invention, the cosmetics usually used for the production of the skin cosmetics together with the anti-inflammatory agents as long as they do not hinder the anti-inflammatory action. Components and other optional substances can be used in combination. Examples of skin cosmetic constituents that can be used in combination are as follows.

  Astringents: citric acid or salt thereof, tartaric acid or salt thereof, lactic acid or salt thereof, zinc sulfate, hypericum extract, dio extract, cornflower extract, maroni extract, salvia extract, melissa extract and the like.

  Bactericidal and antibacterial agents: benzoic acid or salts thereof, paraoxybenzoic acid ester, salicylic acid, sodium salicylate, sorbic acid, resorcin, phenoxyethanol, bisabolol, hinokitiol, menthol, chitosan, chitosan degradation product, yucca extract, aloe extract, caihi extract, etc.

  Whitening agents: ascorbic acid and its derivatives, sulfur, placenta extract, kojic acid and its derivatives, arbitin and its derivatives, glutathione, oil-soluble licorice extract, urgonum extract, nematode extract, Sakuhakuhi extract, mannentake mycelium culture or its extract Products, Gymnema extract, Linden extract, Peach leaf extract, Button pi extract, Clam squirrel extract, Hypericum extract etc.

  Moisturizer: serine, glycine, collagen, hydrolyzed collagen, fibronectin, keratin, royal jelly, glycerophospholipid, glyceroglycolipid, sphingophospholipid, linoleic acid or its ester, pectin, bifidobacteria fermentation product, lactic acid bacteria fermentation product, yeast extract , Soybean phospholipid, γ-oryzanol, yokuinin extract, sage extract, diatom extract, beetle aloe extract, burdock extract, mannen wax extract, wheat bran, rice bran extract and the like.

  Cell Activator: Riboflavin or its derivative, Pantothenic acid or its derivative, Carrot extract, Panax ginseng extract, Loofah extract, Boppi extract, Peonies extract, Mukuroji extract, Safflower extract, Loquat leaf extract, Yukinosita extract, Jasmine extract, Mannen wax extract, etc. .

  Anti-inflammatory / anti-allergic agents: azulene, allantoin, indomethacin, lysozyme chloride, oxybenzone, glycyrrhizic acid or its derivative, glycyrrhetinic acid or its derivative, diphenhydramine hydrochloride, tranexamic acid or its derivative, estradiol, cortisone, hydrocortisone, prednisolone, corticosterone, Arnica extract, Sanshishi extract, Artemisia extract, Senkyu extract, Bow-fu extract, etc.

  Antioxidant and active oxygen scavenger: Propyl gallate, baicalin, baicalein, superoxide dismutase, catalase, ogon extract, age extract, kina extract, perilla extract, thyme extract, caraway extract, nutmeg extract, mace extract, laurel extract, clove extract, Willow tree extract etc.

Hereinafter, the present invention will be described with reference to examples.
Production Example 1
To 100 g of the roughly crushed raw material plant, 2000 ml of an extraction solvent is added, and the soluble components are eluted by maintaining at 70 ° C. for 3 hours with gentle stirring. Then, the extract obtained by filtration is concentrated under reduced pressure at 40 ° C. and dried with a vacuum dryer.

  The above extraction treatment was carried out using 80% water-containing ethanol or ethanol as a solvent, starting from hydrangeas (stalks and leaves), scallops (flowers), mokokuju (bark), salamanders (bark), and kozanreki (bark). The yield of the extract was as shown in Table 1.

Test example 1
The five 80% ethanol extracts obtained in Production Example 1 were tested for phospholipase A2 inhibitory activity by the following method.

Test method: RBL-2H3 cells of rat leukemia cells are cultured in a 75 cm ² flask in MEM medium containing 15% FBS at 37 ° C. under 5% CO ₂ , and the cells are collected by a conventional method. The obtained cells are adjusted to 5 × 10 ⁵ cells / ml in MEM medium containing 15% FBS, and [3H] arachidonic acid (50 μCi / 500 μl) is further added at a rate of 3 μl / 10 ml. 1 ml of the resulting adjustment solution is seeded on a 24 well plate and cultured overnight at 37 ° C. under 5% CO ₂ . Discard the medium in each well, wash with PBS (−), add serum-free MEM medium, and preincubate at 37 ° C. for 30 minutes. Next, a solution in which the sample obtained in Production Example 1 is dissolved is added to the serum-free MEM medium, and preincubation is similarly performed for 10 minutes. Further, 10 μl of 1 mM A23187 is added and incubated at 37 ° C. for 5 minutes. After the reaction, 400 μl of the supernatant is taken under ice cooling, 6 μl of scintillation cocktail is added, and the radioactivity is measured with a liquid scintillation counter. Similarly, a blank test (no A23187 stimulation) and a control (solvent of the sample solution) are measured, and the phospholipase A2 activity inhibition rate is obtained by the following formula.

Inhibition rate (%) = [1− (A−C) / (B−C)] × 100
However,
A: Radioactivity at the time of sample addition B: Radioactivity of control C: Radioactive sample concentration in the blank test is changed stepwise, the above measurement is performed, and the sample concentration at which the inhibition rate is 50% is obtained by interpolation. . The test results are shown in Table 2.

Test example 2
The five types of 80% ethanol extracts obtained in Production Example 1 were tested for hyaluronidase inhibitory activity by the following method.

Test method: A hyaluronidase solution (400 units / ml, pH 9.1 borate buffer) (0.1 ml) and a sample solution (0.2 ml) were mixed, heated to 37 ° C. for 20 minutes, and then an activator solution (0.01 % Compound 48/80, 2.5 mM CaCl ₂ , 0.15 M NaCl) is added and warmed to 37 ° C. for 20 minutes to activate the enzyme. After adding 0.5 ml of potassium hyaluronate buffer and reacting at 37 ° C. for 40 minutes, 0.2 ml of 0.4N sodium hydroxide is added and cooled with ice to stop the reaction. Next, 0.2 ml of 0.8 M boric acid solution (pH 9.1) is added, heated in a boiling water bath for 3 minutes, and immediately cooled on ice for 10 minutes. By adding 6.0 ml of DMAB reagent (10 g of p-dimethylaminobenzaldehyde dissolved in 12.5 ml of 10N hydrochloric acid and 87.5 ml of acetic acid and diluted 10-fold with acetic acid) and keeping at 37 ° C. for 20 minutes The N-acetylglucosamine released by the enzyme reaction is colored and the absorbance at a wavelength of 585 nm is measured. As a control, the same operation and absorbance measurement are performed when water is added instead of the sample solution.

From the above measurement results, the inhibition rate of hyaluronidase activity is calculated by the following formula.
Inhibition rate (%) = [1- (A1-A0) / (A3-A2)] × 100
However,
A0: Absorbance when the sample solution is added; Before the enzyme reaction starts A1: Absorbance when the sample solution is added; After the enzyme reaction ends A2: Control; Before the enzyme reaction starts A3: Control; Concentration of the sample solution after the enzyme reaction ends The above inhibition rate is measured step by step, and the sample concentration at which the inhibition rate is 50% is determined by interpolation. The test results are shown in Table 3.

Test example 3
The five types of 80% ethanol extracts obtained in Production Example 1 were tested for histamine release inhibitory action by the following method (because intracellular histamine was released and hexosaminidase was released at the same time. This is a method for evaluating histamine release inhibitory action using hexosaminidase release as an index.)

Test method: Inoculate 1.0 × 10 ⁶ RBL-2H3 cells in a MEM medium supplemented with 15% FBS in a 25 ml flask, and culture at 37 ° C. for 4 days under 5% CO ₂ . Thereafter, trypsinization and centrifugation (1000 rpm, 2 minutes) are performed to obtain cells as a precipitate. This was suspended in 4.0 × 10 ⁵ cells / ml in the same medium as above, and ⁵ μl of mouse monoclonal anti-dinitrophenyl group IgE (DNP-specific IgE) was added thereto, and the concentration was 0.5 μg / ml. To do. 80 μl of the obtained cell suspension is seeded in a 96-well plate and cultured at 37 ° C. for 24 hours under 5% CO ₂ . After completion of the culture, the medium in each well is removed and washed with a siligarian buffer. Next, 30 μl of the buffer solution and 10 μl of the sample solution are added and incubated at 37 ° C. for 10 minutes. Next, 10 μl of dinitrophenylated bovine serum albumin (DNP-BSA) is added and further incubated at 37 ° C. for 15 minutes. Thereafter, 10 μl of the supernatant is transferred to a new 96-well plate under ice-cooling, and 10 μl of 1 mM p-nitrophenyl-N-acetyl-β-D-glucosamide solution is added thereto and incubated at 37 ° C. for 1 hour. After completion of the _{reaction, 0.1M Na 2 CO 3 · NaHCO} 3 solution 250μl was added to measure the absorbance A at 415nm in a microplate reader. The same treatment and absorbance measurement are performed for the cell suspension without adding the sample solution (the absorbance measured at this time is B). Further, the same treatment and absorbance measurement are performed using the buffer solution instead of the cell suspension (the absorbance measured at this time is C). Then, the hexosaminidase release inhibition rate is calculated by the following formula.

Hexosaminidase release inhibition rate (%) = [1- (A−C) / (B−C)] × 100
The concentration of the sample solution is changed stepwise, the inhibition rate is measured by the above method, and the concentration of the sample solution at which the inhibition rate is 50% is determined by interpolation. The test results are shown in Table 4.

Production Example 2
Add 1000 ml of extraction solvent to 100 g of the coarsely crushed plant material, and maintain at 50 ° C. for 3 hours with gentle stirring. Thereafter, filtration is performed to obtain an extract. The above extraction process was performed using 50% water-containing 1,3-butylene glycol as a raw material, starting from hydrangeas (stalks and leaves), kowakutsuji (bark), mobokuju (bark), salamander (bark), and kozanreki (bark). It was. The solid content yield calculated from the solid content concentration of the obtained extract was as shown in Table 5.

Test example 4
The five types of 50% 1,3-butylene glycol extracts according to Production Example 2 were tested for phospholipase A2 inhibitory action, hyaluronidase inhibitory action, and histamine release inhibitory action in the same manner as in Test Examples 1-3. The results are shown in Tables 6-8.

Test Example 5
A lotion-like coating solution containing one or two of the five 80% ethanol extracts according to Production Example 1 and a control solution having the same composition except that no 80% ethanol extract was contained were prepared. The effect of preventing razor losing was tested. Table 9 shows the composition of each coating solution and the evaluation of the effect of preventing razor losing.

(Note 1) Composition of coating solution: Numerical value is g; The total amount is 100 ml using purified water.
(Note 2) Razor loss prevention test method: 50 male subjects who lose razors are divided into 5 groups of 8 each, and different application liquids are applied to each group after shaving immediately after shaving, and the application effect is as follows. Let us report on the criteria.
Criteria for improving razor loss:
Effective Razor loss disappeared Effective Razor loss weakened Slightly effective Razor loss slightly weakened Invalid No change in razor loss The test criteria below 20% were judged as “invalid” A, 20% or more and less than 50% was evaluated as B, 50% or more and less than 80% was evaluated as C, and 80% or more was evaluated as D.

  In addition, at the same time as the determination on the prevention of razor losing, the impression was given about the degree of irritation to the skin (irritation), but all subjects answered that no irritation was felt.

Example 1
A cream having the following composition, to which an anti-inflammatory action was imparted by adding 80% ethanol extract of Chickpea according to Production Example 1, was produced by a conventional method for producing cream.
Hydrangea extract 1.0% by weight
Stearic acid 2.0
Stearyl alcohol 7.0
Lanolin 2.0
Squalane 5.0
Glycerin monostearate ester 2.0
Polyoxyethylene (25EO) cetyl alcohol ether 3.0
2-Octyldodecyl alcohol 6.0
Propylene glycol 5.0
Ethylparaben 0.3
Perfume appropriate amount Purified water balance

Example 2
A milky lotion having the following composition to which an anti-inflammatory effect was imparted by adding 80% ethanol extract of Kojitsutsugi according to Production Example 1 was produced according to a conventional method for producing a milky lotion.
Kotsutsuji Extract 1.0% by weight
Stearic acid 2.0
Cetyl alcohol 1.5
Vaseline 5.0
Liquid paraffin 10.0
Polyoxyethylene (10EO) oleate 2.0
Polyethylene glycol 1500 3.0
Triethanolamine 1.0
Ethylparaben 0.3
Perfume appropriate amount Purified water balance

Example 3
Three types of 80% ethanol extracts according to Production Example 1 were added to produce a pack having the following composition to which an anti-inflammatory action was imparted.
Moubokuju extract 1.0% by weight
Sarasouju extract 1.0
Kozanreki Extract 2.0
Polyvinyl alcohol 13.0
Ethyl alcohol 7.0
Dipropylene glycol 5.0
Polyoxyethylene (60EO) hydrogenated castor oil 5.0
Olive oil 5.0
Tocopherol acetate 0.2
Ethylparaben 0.2
Perfume

Example 4
A jelly-like cosmetic composition having the following composition to which an anti-inflammatory action was imparted by adding 80% ethanol extract of Production Example 1 was produced.
Sarasoju extract 3.0% by weight
95% ethyl alcohol 10.0
Dipropylene glycol 15.0
Polyoxyethylene (50EO) oleyl alcohol ether 2.0
Carboxyvinyl polymer 0.05
L-Arginine 0.1
Ethylparaben 0.3
Perfume appropriate amount Purified water balance

Claims (2)

  An anti-inflammatory agent which is extracted from the bark of Mobokuju or the bark of Kouzalenki and contains any two or more of a phospholipase A2 inhibitor, a hyaluronidase inhibitor and a histamine release inhibitor as active ingredients.   A skin cosmetic comprising the anti-inflammatory agent according to claim 1.