JP2003277285A - Arginase activity accelerator and skin care preparation containing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an arginase activity accelerator which, even in a small amount, can adjust the arginase activity of the skin and can continuously impart a moisture retention effect to the skin; and a skin care preparation containing the same. <P>SOLUTION: The skin care preparation contains (a) 0.0001-5 wt.% arginase activity accelerator containing an extract of bakumondou (the knots of roots of Ophiopogon japonicus ker-Gawler) and (b) 0.01-50 wt.% 6-22C carboxylic acid or its derivative. <P>COPYRIGHT: (C)2004,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an arginase activity promoter and a skin external preparation containing the same, more specifically, an arginase activity promoter having an excellent effect of exerting a moisturizing effect and having excellent safety and stability. And a skin external preparation such as cosmetics and pharmaceuticals containing the same.

[0002]

2. Description of the Related Art Usually, the skin surface of human beings is covered with a sebum film to appropriately suppress the evaporation of water. It is very important to keep the water content of the skin in an appropriate range from the viewpoint of the health of the skin, and when the water content is insufficient, the skin becomes rough. Therefore, in the "skin external preparation" which is applied to the so-called outer skin such as cosmetics and pharmaceuticals, polyhydric alcohols such as glycerin, 1,3-butylene glycol and sorbitol, pyrrolidone carboxylate and hyalurone are used for prevention and improvement of rough skin. Acid mucopolysaccharides such as acids, chitin, chitosan and their derivatives, protein hydrolysates,
Moisturizing ingredients such as plant extracts and urea have been blended.
However, the methods using these moisturizing components only utilize the physicochemical moisturizing properties of the substance on the skin surface, and are not based on the physiological function of the substance on skin cells. Also, if it is mixed in a large amount, it may have an unpleasant stickiness and the feel may not be preferred. Furthermore, since these moisturizing components lose their effect when they are removed from the skin, the effect must be said to be transient. Therefore, it has been desired to develop a drug that acts on skin cells to promote the production of moisturizing ingredients. Arginase is an enzyme in the urea cycle that hydrolyzes arginine into ornithine and urea, and is widely distributed in the living world including vertebrate liver and kidney. Its presence in human skin has been known for a long time, and is known as an ornithine-supplying enzyme for polyamine biosynthesis and proline biosynthesis associated with epidermal cell proliferation.

In Japanese Patent Laid-Open No. 10-7581,
The physiological function of this arginase in the skin is related to the production of "urea" which is a natural moisturizing factor (NMF) component of the skin, and the extract of "Kizu" as a specific herbal extract extracts the arginase activity in the skin. It is disclosed to regulate and provide a continuous moisturizing effect to the skin. "Urea" is an important ingredient for the skin that not only has a high moisturizing ability but also has a keratolytic and exfoliating action and a keratin softening action, and has been frequently blended in external skin preparations such as cosmetics and pharmaceuticals since ancient times. It is an ingredient. However, since it easily reacts with water and decomposes, and when decomposed, ammonia and carbon dioxide gas are generated, which may cause an odor and cause a safety problem. Furthermore, the external preparation for skin containing urea has a drawback that its use site is limited because it has a peculiar stimulating sensation. On the other hand, by using this arginase activity promoter, it is possible to exert the effectiveness of “urea” itself while solving these problems by increasing the production of “urea” in the skin.
Furthermore, it is considered to be a very useful method because the effect can be expected to be sustainable. Therefore, there has been a demand for the development of an arginase activity promoter which can exert a higher effect with a smaller amount and is excellent in safety and stability.

[0004]

DISCLOSURE OF THE INVENTION The present invention solves the above-mentioned problems, and can control the arginase activity in the skin even in a small amount to continuously give a moisturizing effect to the skin, and further, in terms of safety and stability. An object is to provide an excellent arginase activity promoter.

[0005]

That is, the present invention is as follows.
(1) a. An arginase activity promoter containing the extract of Bakumondou as an active ingredient, (2) a. The arginase activity promoter described in (1) is 0.0 as a dry residue.
001-5% by weight, b. A skin external preparation containing 0.01 to 50% by weight of a carboxylic acid having 6 to 22 carbon atoms or a derivative thereof, (3) b. The carboxylic acid having 6 to 22 carbon atoms contains 80% by weight or more of cis-9-octadecenoic acid, and 85% by weight of cis-9-unsaturated fatty acid.
The external preparation for skin according to (2) above, which comprises the above.

[0006]

BEST MODE FOR CARRYING OUT THE INVENTION The extract of Bakumondou used in the present invention means "snake beard", a perennial plant of the family Liliaceae. Scientific name: Opiopogon japonicus ke
It is an extract extracted from various roots of r-Gawler (crude drug name: Bakumondofu “Bakumondou”) with various solvents. In addition, in this Bakumondou, Ophiopognin A, B,
It contains ingredients such as steroidal saponins such as C, D, deltonin, and ruscogenin, homoisoflavonoids such as ophiopogonanoin A, sterols such as β-sitosterol, and stigmasterol. It is used as a cough suppressant, phlegm remover, nourishing, tonic, diuretic, etc. (Japanese Medicinal Herbs, New Japan Law Publishing). The extract can be extracted by heating under reflux or immersion with one or more solvents selected from hydrocarbons, esters, ketones, ethers, halogenated hydrocarbons, alcohols and water, but preferably water or It is extracted with one or more solvents selected from ethanol, propylene glycol, isopropyl alcohol, and 1,3-butylene glycol, and more preferably extracted with water. The extract of Bakmondou in the present invention means the extraction solution itself or a concentrate thereof.

In the external preparation for skin of the present invention, the content of the extract of Bakmondou as the component a is preferably 0.0001 to 5% by weight as a dry residue in the total amount of the composition, and 0.0005 to 5% by weight. More preferably, it is 3% by weight. If the dry residue is less than 0.0001% by weight, the moisturizing effect and skin roughening improving effect are difficult to be exhibited, and if the dry residue exceeds 5% by weight, stability over time becomes problematic and cost is disadvantageous. However, the dry residue in the present invention refers to a solute which is a residue when the extract is usually dried at 105 ° C. or dried under reduced pressure to remove the solvent, but when the extraction solvent is nonvolatile. In addition, the dissolved mass is obtained as a calculated value from the value obtained by quantifying the amount of solvent by gas chromatography, high performance liquid chromatography, etc., and is regarded as the amount of dry residue.

In the external preparation for skin of the present invention, the carboxylic acid having 6 to 22 carbon atoms or the derivative thereof as the component b is a carboxylic acid having 6 to 22 carbon atoms, which is a straight chain or branched one having 6 to 22 carbon atoms. Divalent or polyvalent carboxylic acids, for example, hexanoic acid, octanoic acid, 1,8-octanedicarboxylic acid, decanoic acid, dodecanoic acid, tetradecanoic acid, c
is-9-octadecenoic acid, cis-9, cis-12
-Octadecadienoic acid, eicosanoic acid, docosanoic acid and the like can be mentioned, and derivatives thereof include esters with monohydric or polyhydric alcohols, amides which are dehydration condensation products with ammonia and organic amines, and the like. Further, alcohols obtained by reducing these carboxylic acids, ethers of the alcohols with monohydric or polyhydric alcohols, inorganic acids, esters with organic acids and the like can be mentioned. However, in the present invention, the component b is a lipophilic component among the above components, specifically, a nonionic surfactant having an HLB of 8 or less or an oil component. In addition, HLB is Griffin (W.
C. Griffin: J. Soc. Cosmetic
Chemists, 33, 1180 (1960)). As the b component, preferably, a carboxylic acid having 6 to 22 carbon atoms is cis-.
80% by weight or more of 9-octadecenoic acid, and ci
A fatty acid or a derivative thereof containing 85% by weight or more of s-9-unsaturated fatty acid. The cis-9-unsaturated fatty acid has a cis-9- double bond in the fatty acid skeleton. However, the fatty acid composition is determined by the area ratio measured by gas chromatography.

In the external preparation for skin of the present invention, the blending amount of the component b is 0.01 to 50% by weight, preferably 0.1 to 30% by weight in the total amount of the composition. b component is 0.01
When the amount is less than 50% by weight, sufficient moisturizing effect and the effect of preventing rough skin cannot be obtained, and when the amount exceeds 50% by weight, the feeling is unpleasant and irritation may occur.

The arginase activity-promoting agent of the present invention can be suitably used by blending it with a skin external preparation, for example, it can be used in cosmetics, pharmaceuticals and the like. In addition, in the present invention, it is possible to mix an additive which is commonly used in external preparations for skin such as cosmetics and pharmaceuticals, within a range that does not impair the performance of the present invention. For example, hydrocarbon-based oils such as liquid paraffin, liquid isoparaffin, squalane, vaseline, and solid paraffin; natural fats and oils such as beef tallow, lard, and fish oil; ester oils; waxes; linear and cyclic dimethylpolysiloxanes and polyether-modified dimethyl Silicone derivatives such as polysiloxane and amino-modified dimethylpolysiloxane;
Oily bases such as ceramide, cholesterol, protein derivative, lanolin, lanolin derivative, lecithin; polyoxyethylene alkyl ether, polyethylene glycol fatty acid ester, polyoxypropylene alkyl ether, polypropylene glycol fatty acid ester, sorbitan fatty acid ester, polyoxyethylene sorbitan fatty acid Nonionic surfactants such as esters, polyoxyethylene hydrogenated castor oil, polyglycerin fatty acid esters, polyoxyethylene glycerin fatty acid esters, glycerin monofatty acid esters, alkyl polyglucosides, polyoxyethylene polyoxypropylene block polymers, alkanolamides; soaps , Alkyl sulfate ester salt, alkyl ether sulfate ester salt, α-olefin sulfonate, acyl Methyl taurate, acyl glutamates, acyl glycine salts, Ashiruzarukoshin salts, acyl isethionates, alkyl ether carboxylates,
Anionic surfactants such as amide ether sulfate ester salt and alkyl phosphate ester salt; amphoteric surfactants such as alkyldimethylaminoacetic acid betaine, amidopropyldimethylaminoacetic acid betaine, amide amino acid salt and alkyliminodiacetate salt; alkylamine Semipolar surfactants such as oxides and polyoxyethylene alkylamine oxides; cationic surfactants such as alkyltrimethylammonium chloride and dialkyldimethylammonium chloride;
Hydrochlorides and acetates of alkylamines and amidoamines; pigments such as inorganic pigments, pearl pigments, metal powder pigments and organic pigments;
Natural pigments such as chlorophyll and β-carotene; water-soluble polymers such as alginic acid, carboxyvinyl polymer, carboxymethylcellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, xanthan gum, hyaluronic acid; magnesium sulfate, sodium chloride, sodium citrate, pyrrolidonecarboxylic acid Inorganic or organic salts such as sodium; acids and alkalis that are pH adjusting agents, bactericides, chelating agents, antioxidants, anti-inflammatory agents,
UV absorbers, natural extracts derived from animals and plants, and fragrances can be added.

[0011]

The present invention will be described in more detail with reference to Examples. 1. Preparation of Bakumondou Extract After drying, add 300 ml of water to 30 g of Bakumondou sliced into 5 mm pieces and heat at 70 ° C. for 3 hours.
Insoluble matter was removed from the extract by filtration, and the obtained filtrate was dried under reduced pressure to give a Bakmondou extract. The dry residue (effective content) in this filtrate was 7.4% by weight. 2. Preparation of tree-pull extract Extract 300 g of water after adding 30 g of tree tree crushed after drying 7
The mixture was heated at 0 ° C. for 3 hours, then insoluble matter was removed from the extract by filtration, and the obtained filtrate was dried under reduced pressure to obtain a wood extract. The dry residue (effective content) in this filtrate was 1.
It was 5% by weight.

Example 1 Arginase activity promoting effect of cultured cells Mouse epidermal cells (PAM212) were seeded in a culture dish having a diameter of 10 cm, and the culture was continued until the culture dish was completely covered with the cells (medium: Dulbecco's medium). Modified Eagle Medium +
Fetal bovine serum 10%). Then, the extract of each plant has a concentration in the medium of 0 to 0.0 as a dry residue of the extract.
It was added so as to be 5% by weight and cultured for 3 days. After culturing, the cells were collected, and the intracellular arginase activity was measured 5 times for each concentration of the extracted extract, and the average value was calculated. To measure the amount of arginase activity in cells,
It was measured using a homogenate homogenized in ml of 25 mM Tris-hydrochloric acid buffer (pH = 7.5). Transfer 0.05 ml of the homogenate to a 1.5 ml stoppered microtube and add 5 ml for activation of arginase.
Heat treatment was performed at 0 ° C. for 10 minutes. Then, 0.04 ml of 0.1 M L-arginine solution was added and the mixture was mixed at 37 ° C. for 3 days.
Incubated for hours. 0.0 after incubation
1 ml of 60% perchloric acid was added and mixed to stop the enzyme reaction, and the mixture was centrifuged at 10,000 G to measure the amount of urea in the obtained supernatant. The intracellular arginase activity amount was relatively expressed by setting the amount of urea produced when the extract of each plant was not added to 100. The results are shown in Table 1. The amount of urea used was Urea Nitrogen-Test Wako manufactured by Wako Pure Chemical Industries, Ltd. according to the instruction manual. In other words, 0.02 ml of the supernatant and 5 ml of the color reagent solution (mixture of color stock solution A and color stock solution B at a ratio of 5: 1) were mixed and the mixture was placed in a boiling water bath to form 2
After heating for 5 minutes, cool in running water and use a spectrophotometer for 530
The absorbance at nm was measured, and the amount of urea in the supernatant was calculated from the calibration curve of urea separately determined.

[0013]

[Table 1]

From Table 1, the extract of Bakumondou was excellent in promoting the arginase activity at the level of cultured cells, and the effect at a low concentration was clearly superior to that of the extract of Kizu.

Example 2 and Comparative Example 1 Effect of Arginase Activity Promoter by Hairless Mouse Application Test The following test was carried out in order to examine whether the same effect was exhibited in the test using animals. 6-week-old male hairless mice purchased from Hoshino Laboratory Animals (1 for each group)
(0 animals) was applied with 200 μL of a 50% ethanol solution containing 0.01% Bakumondou water extract, and this application was performed twice a day continuously for 30 days.
After the application period, the water content of the stratum corneum is changed to SKICON-2.
00 (manufactured by IBS), mouse skin was collected, immersed in 0.24 M ammonium chloride (pH = 9.4) at 0 ° C. for 30 minutes, and then peeled from the dermis using tweezers. Only the epidermis was collected. The collected epidermis was homogenized with 19 times the wet weight of physiological buffered saline and then centrifuged, and the amount of urea and the amount of arginase activity in the supernatant were measured by the same method as in Example 1. In addition, a 50% ethanol-free application group containing no extract and a non-application group were used as comparative examples, and the non-application group was set to 100, and the epidermal stratum corneum water content, the amount of urea in the epidermal homogenate, and the amount of arginase activity in each group were compared. I asked. The results are shown in Table 2.

[0016]

[Table 2]

As is clear from the results in Table 2, the extract of Bakmondou of the present invention increased the arginase activity of the epidermis of hairless mice to increase the amount of urea and the water content of the stratum corneum. Further, in this test, no side effects such as inflammatory hypersensitivity reaction and erythema were observed at the site to which the Bakumondou extract was applied. From this, it was confirmed that the extract of Bakumondou extract effectively promotes the arginase activity within the range where no side effect is exhibited, increases the production of urea amount, and increases the stratum corneum water content.

Examples 3 to 5 and Comparative Example 2: Cream An external preparation for skin, which is a cream shown in Table 3, was prepared and evaluated by the following methods. However, the same extract as that of Example 1 was used as the Bakumondou extract and the wood extract. The results are shown in Table 3. (1) Persistence of moisturizing effect 20 women (21 to 35 years old) were panelists, cream was used after washing the face, and the moisture of the skin after 2 hours was judged as follows, the average of 20 people Seeking the value,
An average value of 1.5 points or more was evaluated as a skin external preparation having a good moisturizing effect. 2 points: When feeling that the skin is sufficiently moisturized just after use. 1 point: When it is felt that the skin is not sufficiently moisturized immediately after use. 0 point: When it feels that the skin does not have enough moisture compared to immediately after use. (2) Skin Roughening Improvement Effect Ten women (aged 25 to 35) with skin roughness were used as panelists, and the condition of the skin when the cream was used twice a day for two consecutive weeks was judged as follows, The average value of 10 persons was obtained, and an average value of 1.5 points or more was evaluated as a skin external preparation having a rough skin improving effect. 2 points: When it is felt that the rough skin has been healed. 1 point: When it is felt that the rough skin has healed slightly. 0 point: When it is felt that there is no skin roughening improving effect. (3) Temporal stability The cream is sealed in a transparent glass container, and the temperature is -5 ° C, 25 ° C.
The state when stored at 45 ° C. for one month was investigated and evaluated according to the following three grades. ◯: Stability is good (no change in appearance and no spots etc.) Δ: Slightly poor stability (slight precipitation or slight separation is observed, or slight spots or lumps are generated, or Slightly colored or discolored.) X: Poor stability (clearly precipitates or separates, or causes lumps or lumps, or clearly colored or discolored).

[0019]

[Table 3]

Note 1; EXTRA OS-85 (manufactured by NOF CORPORATION) <Fatty acid composition: 87% by weight of cis-9-octadecenoic acid, cis-9, 4% by weight of cis-12-octadecadienoic acid, hexadecanoic acid 3% by weight, octadecanoic acid 4% by weight, docosanoic acid 1% by weight, other fatty acids 1% by weight> Note 2; NOFABLE EO-85S (NOF CORPORATION)
<Production of fatty acid: cis-9-octadecenoic acid 86% by weight, cis-9, cis-12-octadecadienoic acid 5% by weight, hexadecanoic acid 3% by weight, octadecanoic acid 4
% By weight, 1% by weight of docosanoic acid, 1% by weight of other fatty acids> From Examples 3 to 5, the external preparation for skin of the present invention was excellent in the durability of the moisturizing effect of the skin and the effect of improving rough skin, and was also excellent in stability. . On the other hand, in Comparative Example 2, since the wood extract was used in place of the Bakumondou extract, the rough skin improving effect was weakened and the stability over time was problematic.

Examples 6 to 8 and Comparative Example 3: Lotion A lotion shown in Table 4 was prepared as a skin external preparation, and evaluations (1) and (2) were carried out by the methods of Examples 3 to 5 and (3). ) The stability with time was evaluated by the following method. However, the same extract as that of Example 1 was used as the Bakumondou extract and the wood extract. The results are shown in Table 4. (3) Temporal stability The lotion was sealed in a transparent glass container and stored at 0 ° C, 25 ° C and 40 ° C for 3 months, and its appearance was observed and evaluated according to the following 3 grades. ◯: Good stability (no change in appearance at any temperature) Δ: Slightly poor stability (somewhat at any temperature, causing precipitation or coloration) X: Poor stability (any one) At a temperature of 1, causing precipitation or separation or marked coloration.)

[0022]

[Table 4]

Note 1; EXTRA OS-85 (manufactured by NOF CORPORATION) <Fatty acid composition: cis-9-octadecenoic acid 87% by weight, cis-9, cis-12-octadecadienoic acid 4% by weight, hexadecanoic acid 3% by weight, octadecanoic acid 4% by weight, docosanoic acid 1% by weight, other fatty acids 1% by weight> Note 2; NOFABLE EO-85S (NOF CORPORATION)
<Production of fatty acid: cis-9-octadecenoic acid 86% by weight, cis-9, cis-12-octadecadienoic acid 5% by weight, hexadecanoic acid 3% by weight, octadecanoic acid 4
% By weight, 1% by weight of docosanoic acid, and 1% by weight of other fatty acids> From Examples 6 to 8, the external preparation for skin of the present invention was excellent in the durability of skin moisturizing effect and the effect of improving rough skin, and also excellent in stability. . On the other hand, in Comparative Example 3, the wood extract was used in place of the Bakumondou extract, so that the moisturizing effect persistence and the rough skin improving effect were weakened.

[0024]

INDUSTRIAL APPLICABILITY The arginase activity promoter of the present invention can control the arginase activity in the skin even in a small amount to continuously give a moisturizing effect to the skin, and is also excellent in safety and stability.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61P 43/00 111 A61P 43/00 111 F term (reference) 4C083 AA082 AA111 AA112 AC022 AC072 AC102 AC122 AC172 AC231 AC251 AC252 AC352 AC402 AC422 AC432 AC442 AC482 AD152 AD662 CC02 CC04 CC05 EE12 4C088 AB85 AC11 CA05 CA06 MA02 MA63 NA14 ZA89 ZC19

Claims (3)

[Claims] 1. A. An arginase activity promoter containing the extract of Bakumondou as an active ingredient. 2. A. 0.0001-5% by weight of the arginase activity promoter according to claim 1 as a dry residue, b.
A carboxylic acid having 6 to 22 carbon atoms or its derivative is 0.0
A skin external preparation characterized by containing 1 to 50% by weight. 3. b. Carboxylic acid having 6 to 22 carbon atoms is ci
The external preparation for skin according to claim 2, which contains 80% by weight or more of s-9-octadecenoic acid and 85% by weight or more of cis-9-unsaturated fatty acid.