JP2002029991A - Arginase activity accelerator and preparation for external use for skin containing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an arginase activity accelerator capable of controlling the arginase activity in the skin and continuously imparting humectant effect to the skin even with a small amount, and excellent in safety, and to provide a preparation for external use for the skin containing the arginase activity accelerator and excellent in the durability of the humectant effect, the improving effect on the improvement of rough skin and safety. SOLUTION: An arginase activity accelerator containing an extract of GOSYUYU (fruit of Evodia rutaecarpa) as an active ingredient.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an arginase activity promoter and an external preparation for skin containing the same, and more particularly, to an arginase activity promoter excellent in the effect of exhibiting a moisturizing effect and excellent in safety, and the same. The present invention relates to an external preparation for skin such as cosmetics and medicines contained therein.

[0002]

2. Description of the Related Art Normally, the surface of human skin is covered with a sebum film, and the evaporation of water is appropriately suppressed. It is very important to keep skin moisture in an appropriate range from the viewpoint of skin health, and lack of moisture easily causes skin roughness and the like. Therefore, in "skin external preparations" adapted to the so-called outer skin of cosmetics and pharmaceuticals, polyhydric alcohols such as glycerin, 1,3-butylene glycol and sorbitol, and pyrrolidone carboxylate are used to prevent rough skin and improve rough skin. , Acidic mucopolysaccharides such as hyaluronic acid,
Mixing of moisturizing components such as chitin, chitosan and their derivatives, protein hydrolysates, plant extracts, and urea has been performed. 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. In addition, when a large amount is blended, it may have unpleasant stickiness and the feeling may not be preferred. Furthermore, the effect of these moisturizing components disappears when removed from the skin, so the effect must be said to be transient. Therefore, the development of a drug that works on skin cells to promote the production of moisturizing components has been desired. Arginase is an enzyme in the urea cycle that hydrolyzes arginine to ornithine and urea, and is widely distributed in the living world, including vertebrate liver and kidney. Its existence in human skin has been known for a long time, and is known as an ornithine-supplying enzyme for polyamine biosynthesis and proline biosynthesis related to epidermal cell proliferation.

In Japanese Patent Application Laid-Open No. 10-7581, the physiological function of arginase in the skin is related to the production of "urea" which is a natural moisturizing factor (NMF) component of the skin. It has been reported that Tong's extracted extract modulates arginase activity in the skin to provide a continuous moisturizing effect on the skin. "Urea" is an important component for skin that not only has a high moisturizing ability but also has a keratolytic and exfoliating action and a keratin softening action, and is an ingredient that has long been used in skin and external preparations such as cosmetics and pharmaceuticals. It is. However, it is easily decomposed by reacting with water, and when decomposed, produces ammonia and carbon dioxide gas, so that it is easy to cause problems in odor and safety. Further, since the skin external preparation containing urea has a specific irritating feeling, it has a drawback that its use site is limited. On the other hand, when this arginase activity promoter is used, it is possible to exhibit the effectiveness of "urea" itself while solving these problems by increasing the production of "urea" in the skin. It is considered to be a very useful tool because of its sustainability. Therefore, there has been a demand for the development of a high arginase activity promoter which can exert a higher effect with a smaller amount and which is excellent in safety and stability.

[0004]

DISCLOSURE OF THE INVENTION The present invention solves the above-mentioned problems, and can regulate the arginase activity in the skin even in a small amount to give a continuous moisturizing effect to the skin, and is excellent in safety. An object of the present invention is to provide an accelerator and a skin external preparation containing the same, which has excellent moisturizing effect, skin roughness improving effect, and stability.

[0005]

That is, the present invention provides:
(1) a. Arginase activity promoter containing an extract of Goshuyu as an active ingredient, (2) a.
0.001 to 5% by weight of the arginase activity promoter described in (1) as a dry residue, b. An external preparation for skin, comprising 0.01 to 50% by weight of a carboxylic acid having 6 to 22 carbon atoms or a derivative thereof, (3) b. Carbon number 6 ~
Cis-9- as a carboxylic acid or a derivative thereof 22
Octadecenoic acid is 80% by weight or more and cis-9
-The external preparation for skin according to (2), which contains a fatty acid having an unsaturated fatty acid content of 85% by weight or more or a derivative thereof.

[0006]

BEST MODE FOR CARRYING OUT THE INVENTION The extract of Goshuyu used in the present invention is a plant of the family Rutaceae "Goshuyu" Scientific name: Evodia rutaecarpa
Or "Hongoshuyu" Scientific name: Evodia offi
It is an extract obtained by extracting the immature fruit of C. cinalis as it is or after drying it with various solvents. In addition, this goshuyu contains alkaloids such as evodiamine, dehydrobodiamine, isoevodiamine, and ritaecarpine, ocimene,
It contains essential oils such as Eboden, and is used in Chinese medicine for stomach stomach, diuretics, headache due to water poisoning, vomiting, chest stomach, etc. As a method for extracting Goshuyu extract, hydrocarbons, esters, ketones,
Extraction is performed by heating under reflux or immersion together with one or more solvents selected from ethers, halogenated hydrocarbons, alcohols and water, preferably water or ethanol, propylene glycol, isopropyl alcohol, 1,3-butylene glycol. And extracted with one or more solvents, and more preferably extracted with water. In addition, the extract of goshuyu of the present invention refers to an extract solution itself or a concentrate thereof.

In the external preparation for skin of the present invention, a. The amount of the extract of goshuyu, which is a component, is preferably 0.0001 to 5% by weight, more preferably 0.0005 to 3% by weight, as a dry residue in the total amount of the composition. If it is less than 0.0001% by weight, the moisturizing effect, the effect of improving skin roughness and the effect of improving odor stability are hardly exhibited, and if it exceeds 5% by weight, there is a problem in stability over time and the cost is disadvantageous. The dry residue referred to in the present invention generally refers to a solute that is a residue obtained by drying the extract at 105 ° C. or drying under reduced pressure to remove the solvent. The dissolved mass is determined as a calculated value from the value obtained by quantifying the amount of the solvent by chromatography, high performance liquid chromatography, and the like, and is regarded as the amount of the dried residue.

B. In the carboxylic acid having 6 to 22 carbon atoms as a component or the derivative thereof, the carboxylic acid having 6 to 22 carbon atoms is a linear or branched monovalent or polyvalent carboxylic acid having 6 to 22 carbon atoms, for example, hexane Acid, octanoic acid, 1,8-octanedicarboxylic acid, decanoic acid, dodecanoic acid, tetradecanoic acid, cis-9-octadecenoic acid,
cis-9, cis-12-octadecadienoic acid, eicosanoic acid, docosanoic acid and the like, and derivatives thereof are esters with monohydric or polyhydric alcohols and amides which are dehydration condensates with ammonia or organic amines. Etc.
Further, alcohols obtained by reducing these carboxylic acids, ethers of the alcohols with monohydric or polyhydric alcohols, esters of inorganic acids, organic acids and the like can be mentioned. However, in the present invention, b. The component is a lipophilic component among the above components, specifically, a nonionic surfactant or an oil component having an HLB of 8 or less. b. Preferred as the component is a fatty acid or a derivative thereof in which the carboxylic acid having 6 to 22 carbon atoms or a derivative thereof is cis-9-octadecenoic acid 80% by weight and cis-9-unsaturated fatty acid 85% by weight, and Preferred is 85% by weight of cis-9-octadecenoic acid and c
Fatty acid which is 90% by weight of is-9-unsaturated fatty acid or a derivative thereof. The cis-9-unsaturated fatty acid includes a cis-9-double bond in the skeleton of the fatty acid composition. The fatty acid composition is determined from the area ratio determined by gas chromatography.

In the external preparation for skin of the present invention, b. The compounding amount of the carboxylic acid having 6 to 22 carbon atoms or a derivative thereof as a component is 0.01 to 50% by weight, preferably 0.1 to 30% by weight based on the total amount of the composition. If the amount is less than 0.01% by weight, sufficient durability of the moisturizing effect and the effect of preventing rough skin cannot be obtained, and if the amount exceeds 50% by weight, the feeling becomes poor or irritating, which is not preferable.

The arginase activity promoter of the present invention can be suitably used by blending it with an external preparation for skin.
It can be used for pharmaceuticals and the like.

In the present invention, additives commonly used in external preparations for skin, such as cosmetics and pharmaceuticals, can be blended as long as the performance of the present invention is not impaired. For example, liquid paraffin, liquid isoparaffin, squalane, petrolatum, hydrocarbon oils such as solid paraffin, natural fats and oils such as tallow, lard, fish oil, ester oil, wax, linear and cyclic dimethylpolysiloxane, polyether-modified dimethyl Silicone derivatives such as polysiloxane and amino-modified dimethylpolysiloxane, ceramides, cholesterol, protein derivatives, lanolin, lanolin derivatives, oily bases such as 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 ester, polyoxyethylene hardened castor oil, polyglycerin fatty acid ester, polyoxyethylene Nonionic surfactants such as glycerin fatty acid ester, alkyl polyglucoside, polyoxyethylene polyoxypropylene block polymer, alkanolamide, soap, alkyl sulfate, alkyl ether sulfate, α-olefin sulfonate, acylmethyl Taurine salt, acyl glutamate, acyl glycine salt, acyl sarcosine salt,
Anionic surfactants such as acyl isethionate, alkyl ether carboxylate, amide ether sulfate, and alkyl phosphate, alkyl dimethyl amino acetate betaine, amido propyl dimethyl amino acetate betaine, amide amino acid salt, alkyl iminodiacetic acid Amphoteric surfactants such as salts; semi-polar surfactants such as alkylamine oxides and polyoxyethylene alkylamine oxides; cationic surfactants such as alkyltrimethylammonium chloride and dialkyldimethylammonium chloride; alkylamines and amidoamines Hydrochloride or acetate, inorganic pigment, pearl pigment, metal powder pigment, organic pigment, pigment such as zirconium, natural pigment such as chlorophyll, β-carotene, alginic acid, carboxyvinyl polymer, carboxymethyl Loin, hydroxypropylmethyl cellulose, hydroxyethyl cellulose, xanthan gum, water-soluble polymers, magnesium sulfate or the like hyaluronic acid, sodium chloride, sodium citrate, inorganic or organic salts such as sodium pyrrolidone carboxylic acid,
Acids and alkalis as pH adjusters, bactericides, chelating agents, antioxidants, anti-inflammatory agents, ultraviolet absorbers, natural extracts derived from animals and plants, fragrances and the like can be added.

[0012]

Next, the present invention will be described in more detail by way of examples. (Preparation of Goshuyu extract extract) Water-extracted extract To 30 g of Goshuyu dried and ground, 300 mL of water was added, and the mixture was heated at 70 ° C. for 3 hours. Thereafter, insoluble materials were removed from the extract by filtration, and the obtained filtrate was dried under reduced pressure to obtain Goshuyu. An aqueous extract was obtained. 2. Extract Extract with Ethanol Aqueous Solution To 30 g of Goshuyu dried and ground, 300 ml of ethanol was added, and the mixture was allowed to stand for 4 days and filtered. Next, 300 ml of water was added to the goshuyu swollen with ethanol, and the mixture was heated at 70 ° C. for 3 hours. After that, unnecessary substances were removed from the extract by filtration, and the obtained filtrate was dried under reduced pressure to dryness. An aqueous extract of Goshuyu ethanol was obtained. Hereinafter, the extract extracted by this method is referred to as a water / EtOH extract. (Preparation of Kidori Extract) Water-extracted extract After drying, 300 ml of water was added to 30 g of the pulverized tree and heated at 70 ° C. for 3 hours. Thereafter, the insoluble solution was removed from the extract by filtration, and the obtained filtrate was dried under reduced pressure to obtain a solid. The obtained extract was obtained.

Example 1 Effect of Promoting Arginase Activity by Cultured Cells Mouse epidermal cells (PAM212) were sown in a culture dish having a diameter of 10 cm, and culturing was continued until the culture dish was completely covered with the cells (medium: Dulbecco) Denatured Eagle medium +
Fetal calf serum 10%). Thereafter, the extract of each plant was added to a medium (medium: Dulbecco's modified Eagle medium + fetal calf serum 1).
%) 0-0.05 as the dry residue of the extracted extract
%, And cultured for 3 days. After culture
The medium and cells were collected, and the amount of urea generated by the reaction of arginase in the medium and the amount of arginase activity in the cells were measured. Each concentration point was performed by 5 points. The measurement of the amount of arginase activity in cells was performed by measuring
The measurement was performed using a homogenate homogenized in a mM Tris-HCl buffer (pH = 7.5). 0.05 ml of the homogenate was transferred to a 1.5 mL stoppered microtube, 0.04 mL of a 0.1 M L-arginine solution was added, and the mixture was incubated at 37 ° C. for 3 hours. After the incubation was completed, 0.01 mL of 60% perchloric acid was added and mixed to stop the enzyme reaction, followed by centrifugation at 10,000 G. The amount of urea in the obtained supernatant was measured in the same manner as the amount of urea in the medium. The amount producing 1 micromol of urea per hour was defined as 1 unit of arginase. The urea content in the medium and the intracellular arginase activity were both taken as 100 and the average value of 5 points was calculated. Table 1 shows the results.

[0014]

[Table 1]

From Table 1, it can be seen that the extract of Goshuyu was excellent in promoting arginase activity at the level of cultured cells, and the effect at low concentrations was clearly superior to the Kindori extract. In particular, the water-extracted extract was highly effective.

Example 2 and Comparative Example 1 Effect of Arginase Activity Promoter by Hairless Mouse Application Test Next, the following test was carried out to examine whether or not the same effect is exhibited in a test using animals. Was. Using a 6-week-old male hairless mouse (10 mice per group) purchased from Hoshino experimental animals, 200 μL of a 50% ethanol solution containing 0.01% Goshuyu water extract was applied, and then this application was performed twice a day. For 30 consecutive days. After the application period, after measuring the water content of the skin surface layer using SKICON-200 (manufactured by IBS), mouse skin was collected and placed in 0.24 M ammonium chloride (pH = 9.4) at 0 ° C. for 30 minutes. After soaking for minutes, the skin was separated from the dermis using forceps and only the epidermis was collected. Remove the collected epidermis to the wet weight of 1
After homogenization with 9 times the volume of physiological buffer saline, centrifugation was performed, and the urea content and arginase activity content of the supernatant were measured. The amount of urea is urea nitrogen manufactured by Wako Pure Chemical Industries, Ltd.
It was used according to the instruction manual using Test Wako. Mix 0.02 mL of the medium and 5 mL of the color reagent (a mixture of the color developer A and the color developer B at a ratio of 5: 1) and mix them in a boiling water bath.
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 medium was calculated from a separately determined urea calibration curve. The measurement of the amount of arginase activity was performed in the same manner as the measurement of arginase activity of the cultured cells. Similarly, a group to which a 50% ethanol solution containing 0.01% Goshuyu water / EtOH extract was applied was also tested. Further, a group of application of only 50% ethanol containing no extract extract and a group of only sham operation were used as comparative examples. I asked for the quantity. Table 2 shows the results.

[0017]

[Table 2]

As is evident from the results in Table 2, the extract of Goshuyu of the present invention increased the arginase activity in the epidermis of the hairless mouse, increased the amount of urea, and increased the water content of the stratum corneum. In this test, no side effects such as the occurrence of an inflammatory hypersensitivity reaction, for example, the occurrence of erythema, etc. were observed at the site to which the extract of Goshuyu was applied. From these results, it was confirmed that the extract of Goshuyu effectively promotes arginase activity within a range that does not exhibit any side effects, increases the production of urea, and increases the water content of the stratum corneum.

Examples 3 to 5 and Comparative Example 2 Skin external preparations as creams shown in Table 3 were prepared and evaluated by the following methods. Table 3 shows the results. (1) Persistence of moisturizing effect Twenty women (21 to 35 years old) were used as panelists, and after washing their face, using a topical skin preparation, the moisture of the skin after 2 hours was determined as follows, and 20 women were evaluated. The average of 1.5 points or more was evaluated as a cosmetic having good moisturizing effect and good durability. 2 points: When the skin feels sufficiently moist as in immediately after use. 1 point: When feeling that the moisture of the skin is slightly less than immediately after use. 0 point: when the skin feels less moisture than immediately after use. (2) Skin roughening improvement effect Ten women (25 to 35 years old) who had skin roughening were used as panelists, and the skin condition was determined as follows when skin external preparations were used twice a day for two consecutive weeks. The average value of 10 persons was determined, and an average value of 1.5 or more was evaluated as a cosmetic having an effect of improving skin roughness. 2 points: When the user feels that the rough skin has clearly recovered. 1 point: When the user feels that the rough skin has healed a little. 0 point: When it is felt that there is no effect of improving skin roughness. (3) Stability over time A skin external preparation is sealed in a transparent glass container, and stored at -5 ° C and 25 ° C.
And the condition when stored at 40 ° C for 3 months,
The evaluation was performed in three stages shown below. ：: good stability (no change in appearance and no blemishes at any temperature) △: somewhat poor stability (slight precipitation or slight separation at any temperature) ×: Poor stability (obviously precipitates or separates at any temperature, or produces dots or lumps.) (4) Odor stability Transparent glass for skin external preparation Seal in a container, -5 ℃, 25 ℃
The change in odor after storage for 3 months at 40 ° C. and 40 ° C. was investigated, and evaluated in the following three grades. ：: Good stability (no change in odor) Δ: Slightly poor stability (Slightly changed odor and slightly off-flavor.) ×: Poor stability (Clearly changed odor and off-odor) Has occurred.)

[0020]

[Table 3]

Note 1: EXTRA OS-85 (manufactured by NOF Corporation) <Fatty acid composition: cis-9-hexadecenoic acid 1% by weight, cis-9-octadecenoic acid 87% by weight, c
is-9, cis-12-octadecenoic acid 3% by weight, hexadecanoic acid 3% by weight, octadecanoic acid 6% by weight> Note 2: NOFABLE EO-85S (NOF Corporation)
Fatty acid composition: cis-9-hexadecenoic acid 1% by weight, cis-9-octadecenoic acid 88% by weight, cis-
9, 2% by weight of cis-12-octadecenoic acid, 4% by weight of hexadecanoic acid, 5% by weight of octadecanoic acid> According to Examples 3 to 5, the external preparation for skin of the present invention has a long-lasting moisturizing effect on the skin and an effect of improving rough skin. Excellent as well as excellent stability. In Comparative Example 2, the effect of improving skin roughness, the stability over time, and the odor stability were deteriorated due to the inclusion of the extract of Kodori instead of the extract of Goshuyu.

Examples 6 to 8 and Comparative Example 3 Skin external preparations as lotions shown in Table 4 were prepared, and evaluations (1), (2) and (4) were performed according to the methods of Examples 3 to 5, and (3) The stability over time was evaluated by the following method. Table 4 shows the results. (3) Stability over time 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 three grades. ：: Good stability (no change in appearance at any temperature) Δ: Slightly poor stability (slightly at any temperature, causing precipitation or slight coloring) ×: Poor stability (any At this temperature, precipitates or separates, or is markedly colored.)

[0023]

[Table 4]

Note 1: EXTRA OS-85 (manufactured by NOF Corporation) <fatty acid composition: cis-9-hexadecenoic acid 1% by weight, cis-9-octadecenoic acid 87% by weight, c
is-9, cis-12-octadecenoic acid 3% by weight, hexadecanoic acid 3% by weight, octadecanoic acid 6% by weight> Note 2: NOFABLE EO-85S (NOF Corporation)
Fatty acid composition: cis-9-hexadecenoic acid 1% by weight, cis-9-octadecenoic acid 88% by weight, cis-
9, 2% by weight of cis-12-octadecenoic acid, 4% by weight of hexadecanoic acid, 5% by weight of octadecanoic acid> From Examples 6 to 8, the external preparation for skin of the present invention has a long-lasting moisturizing effect and an effect of improving skin roughness. Excellent as well as excellent stability. In Comparative Example 3, the skin roughness improving effect and the odor stability are deteriorated because the extract of Kodori was added instead of the extract of Goshuyu.

[0025]

It has been confirmed that the arginase activity promoter of the present invention effectively promotes arginase activity even in a small amount, increases the production of urea, and increases the water content of the stratum corneum. In addition, the skin external preparation of the present invention using the same is excellent in persistence of the moisturizing effect on the skin and in improving the roughness of the skin and also in safety and stability.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61P 43/00 107 A61P 43/00 107 111 111 F F-term (Reference) 4C083 AA082 AA111 AA112 AA122 AC022 AC072 AC102 AC122 AC172 AC241 AC251 AC252 AC352 AC422 AC442 AC482 AD112 AD152 AD662 CC02 CC04 CC05 DD23 DD27 DD31 EE12 4C088 AB62 AC04 BA08 BA09 BA10 CA03 CA05 CA06 CA07 MA17 MA28 MA63 NA14 ZA89 ZB22 ZC19

Claims (3)

[Claims] 1. A method comprising: a. An arginase activity promoter containing an extract of goshuyu as an active ingredient. 2. A method comprising: a. 0.0001 to 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 a derivative thereof is 0.0
An external preparation for skin, comprising 1 to 50% by weight. B. 3. A carboxylic acid having 6 to 22 carbon atoms or a derivative thereof containing a fatty acid or a derivative thereof in which cis-9-octadecenoic acid is 80% by weight or more and cis-9-unsaturated fatty acid is 85% by weight or more. The topical skin preparation according to the above.