JPH107581A - Arginase activity promoter - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an arginase activity promoter capable of providing the skin with moisture and luster. SOLUTION: This promoter comprises an extracted essence of dried Akebia qunata as an active ingredient. The arginase activity promoter in the form of preparation for external use for skin is capable of increasing the water content in a keratin layer, excellent in moisture retaining effects on the skin, useful for chapped skin and high in safety.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD [0001] The present invention relates to an arginase activity promoter containing an extract of Kindori as an active ingredient,
In particular, the present invention relates to a skin external preparation excellent in the effect of producing moisturizing substances that retain moisture in the skin by skin cells to give the skin moisture and gloss.

[0002]

BACKGROUND OF THE INVENTION Arginase is an enzyme in the urea cycle that hydrolyzes arginine into ornithine and urea.
It is widely distributed in the biological world, including vertebrate liver and kidney. Its existence has been known in human skin for a long time, and arginase is known as an ornithine supplying enzyme for polyamine biosynthesis and proline biosynthesis related to the proliferation of epidermal cells. However, no report has been found that relates the physiological function of this enzyme in the skin to the production of moisturizing factors, and there is no known drug that regulates arginase activity in the skin to give a moisturizing effect to the skin. . One of the major purposes of skin external preparations such as cosmetics is to prevent and improve skin roughness, but conventionally, various moisturizing substances have been blended to achieve this purpose. However, these moisturizers only rely on the physicochemical moisturizing properties of the substance on the skin surface, not on the physiological function of the substance on skin cells. . In addition, since such moisturizing substances lose their effects when removed from the skin, their effects must be said to be transient. Therefore, there has been a demand for the development of a drug that acts on skin cells to promote the production of moisturizing substances. On the other hand, Mokutsu (Mokutsu) is a crude drug that has been used as a component of Chinese herbal medicine since ancient times, which is obtained by collecting vine stems of akebi or other congener plants, cutting them into rings, and drying them. As a function of Kodori,
Effective in preventing stress stomach ulcers.
Page (1975)｝ Diuretic effect ｛Chem. Phar. Bu
ll., vol. 27, p. 1644 (1979), which has an anti-inflammatory effect, announced at the 90th Annual Meeting of the Pharmaceutical Society of Japan (1970). However, Kindori increases arginase activity,
Further, it has not been known at all that a skin external preparation containing Kinatsu has an effect of increasing urea production in the skin and enhancing skin moisturizing ability.

[0003]

SUMMARY OF THE INVENTION Accordingly, an object of the present invention is to provide an arginase activity promoter containing an extract of Kinatsu as an active ingredient, and a moisturizing substance originally possessed by skin by promoting arginase activity of skin cells. In the form of an external preparation for skin having an excellent effect of moisturizing and lubricating the skin by increasing the skin.

[0004]

Means for Solving the Problems In view of such circumstances, the present inventors have intensively studied the mechanism of producing moisturizing components in skin in order to improve the inherent moisturizing properties of skin. As a result, the function of arginase in the epidermis is
It was found that urea, which is a moisturizing factor expressed during the differentiation process of skin cells, was mainly involved in cell proliferation, not mainly cell proliferation. Based on this finding, the present inventors continued to search for a drug that induces arginase present in epidermal cells, thereby increasing the production of urea as a skin moisturizing factor. The present inventors have found that arginase is induced in the dermatitis, and by adding it to an external preparation for skin, they have succeeded in obtaining an arginase activity promoter which can achieve the above object, and have completed the present invention. That is, the present invention provides an arginase activity enhancer containing an extract of Kinatsu as an active ingredient, and a skin external preparation.

[0005]

BEST MODE FOR CARRYING OUT THE INVENTION The extract of wood mush according to the present invention refers to a wood mock extracted from one of hydrocarbons, esters, ketones, ethers, halogenated hydrocarbons, alcohols and water. It refers to one obtained by heating or refluxing or dipping together with a seed or two or more kinds and filtering. Preferably, the extract of Kinatsu according to the present invention is obtained by adding 300 ml of water to 30 g of dry and pulverized Kinatsu, heating to 70 ° C. and refluxing for 3 hours while refluxing. Say. At this time, the obtained extract may be concentrated and used as a dry solid, or a fraction obtained by further separating and purifying the component having the above effect may be used.

In the present invention, the amount of the extract of the Kimitsu extract in the total amount of the external preparation for skin is 0.0001 to 0.0001 as a dry solid.
It is preferably 10% by weight, particularly preferably in the range of 0.01 to 5% by weight. If the amount is less than 0.0001% by weight, the effect of increasing the arginase activity to produce a moisturizing component may not be sufficiently achieved, and if the amount exceeds 10% by weight, the effect cannot be expected to be increased.

[0007] The external preparation for skin of the present invention refers to those applied to the outer skin such as cosmetics, pharmaceuticals, and quasi-drugs. Therefore, the dosage form is also aqueous, solubilizing, emulsifying, powder, oil-liquid. , Gel, ointment, water-oil two-layer system, water-oil-powder three-layer system. The external preparation for skin of the present invention includes aqueous components, powders, surfactants, oils, humectants, alcohols, pH regulators, preservatives, which are commonly used in external preparations for skin such as cosmetics and pharmaceuticals, in addition to the extract of wood through the skin. , An antioxidant, a thickener, a dye, a fragrance, and the like, if necessary.

Further, the external preparation for skin of the present invention may further contain a known agent if necessary. As these drugs, for example, ascorbic acid derivatives, placenta extract, drugs having a whitening effect such as glutathione, glycerin, drugs having a moisturizing effect such as sorbitol, glycyrrhetinic acid derivatives, anti-inflammatory agents such as indomethacin,
Extracts such as aloe, luffa, and lily; activators such as biotin and pantothenic acid; and blood circulation promoters such as vitamin E derivatives and nicotinic acid derivatives, and the like. The effect can give the skin moist and shiny.

[0009]

The present invention will be described in more detail with reference to the following examples, which should not be construed as limiting the invention. Example 1: An arginase activity enhancer, a wood extract extract, a wood extract , was obtained from Kanai Tokichi Shoten Co., Ltd. (1-9-11 Kaji-cho, Chiyoda-ku, Tokyo). After drying, 300 ml of water was added to 30 g of the pulverized trees and heated at 70 ° C. for 3 hours. Thereafter, insolubles were removed from the extract by filtration, and the obtained filtrate was dried under reduced pressure to obtain an extract of Kinatsu extract.

Evaluation as an Arginase Activity Promoter The arginase activity promoting action was measured using the extract of Kindori obtained by the above procedure. Human epidermal cells were seeded in a culture dish having a diameter of 10 cm, and the culture was continued until the inside of the culture dish was completely covered with the cells. After that, the concentration of the extract of Kodori in the culture medium was 0 to 0 (as dry weight of the extract).
It was added to 0.01% by weight and cultured for 4 days. After the culture, the medium and the 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 is 5
We went point by point. The amount of urea in the medium was measured by using urea nitrogen-test Wako manufactured by Wako Pure Chemical Industries, Ltd. according to the instruction manual. Mix 0.02 ml of medium and 5.0 ml of a coloring reagent (a mixture of a coloring stock solution A and a coloring stock solution B at 5: 1), heat in a boiling water bath for 25 minutes, cool in running water and use a spectrophotometer. The absorbance at 530 nm was measured, and the amount of urea in the medium was calculated from a separately determined urea calibration curve.

The amount of arginase activity in the cells was measured using a homogenate obtained by homogenizing the cells in 1 ml of 25 mM Tris-HCl buffer pH 7.5. Transfer 0.05 ml of the homogenate to a 1.5 ml stoppered microtube and incubate at 50 ° C. for activation of arginase at 10 ° C.
Heat treatment was performed for minutes. 0.04 ml of a 0.1 M arginine solution was added and incubated at 37 ° C. for 30 minutes. After the incubation was completed, 0.01 ml of 60% perchloric acid was added and mixed to stop the enzyme reaction.
Centrifugation was performed at 0 g, and the amount of urea in the obtained supernatant was measured in the same manner as the amount of urea in the medium described above.
Here, the amount that produced 1 micromol of urea per hour was defined as 1 unit of arginase. Urea content in the medium,
Arginase activity in cells is 100
Table 1 shows the average values of the five points.

Table 1 Amount of urea in the culture medium of the extract of Kinatsu Extraction concentration in the intracellular arginase medium (%) Active amount 0 100 100 0.0001 103 103 0.0005 150 253 0.001 281 352 0.01 303 384 0. 05 354 486

The results show that the arginase activity promoter of the present invention is excellent in the arginase activity promoting effect.

Example 2: By hairless mouse application test
Effect of Arginase Activity Promoter As described above, it is clear that the extract of Mokutsu as an arginase activity promoter promotes arginase activity at the cultured cell level and increases the amount of urea produced. The following test was conducted to determine whether the same effect was exhibited in the test.

Using a 6-week-old male hairless mouse (10 mice in each group) purchased from a Hoshino experimental animal, 200 μl of a 50% ethanol solution containing 0.01% extract of wood through was applied, and then this application was performed for 1 day. The test was performed twice consecutively for 30 days. After the application period was completed, the skin stratum corneum moisture content was measured using SKICON-200, and the mouse skin was collected and placed in 0.24 M ammonium chloride pH 9.4 at 0 ° C.
After soaking for 0 minutes, the dermis was peeled off using tweezers, and only the epidermis was collected. The collected epidermis was homogenized with 19 times the wet weight of physiological buffered saline, followed by centrifugation, and the urea content and arginase activity content of the supernatant were measured. The amount of urea was measured by the above-described method of measuring the amount of urea in the medium, and the amount of arginase activity was measured by the same method as the above-described method of measuring the amount of arginase activity of the cultured cells. A group to which only 50% ethanol containing no extract was applied (Comparative Example 1) and a group to which only the simulated operation was applied (Comparative Example 2) were used as Comparative Examples. Table 2 shows the water content of the epidermal stratum corneum, the amount of urea in the epidermal homogenate, and the amount of arginase activity in each group, with the simulated application group taken as 100. Table 2 Example 2 Comparative Example 1 Comparative Example 2 Skin Epidermis Stratum Corneum Water Content 148 105 100 Urea Content in Epidermis 175 103 100 Arginase Activity Content in Epidermis 154 98 100

As is clear from the results in Table 2, it is clear that the arginase activity enhancer of the present invention increases arginase activity in hairless mouse epidermis, increases urea content, and increases water content of the stratum corneum. became. In the test, the site to which the arginase activity promoter according to the present invention was applied had no side effects, such as the occurrence of an inflammatory hypersensitivity reaction or erythema, and therefore, the arginase activity promoter according to the present invention was not observed. However, it was confirmed that arginase could effectively promote arginase activity, increase the production of urea, and increase the water content of the stratum corneum within a range not causing side effects.

In the following Examples 3 to 6, the present invention is applied.
An example of the formulation of a skin external preparation is given below. Example 3 (ointment) Component Parts by weight A Kodori Extract Extract 0.001 White Vaseline 25 Stearyl Alcohol 22 B Propylene Glycol 12 Sodium Lauryl Sulfate 1.5 Preservative / Antioxidant Appropriate Amount Perfume Appropriate Purified Water Appropriate Amount ──────────成分 Dissolve the components belonging to the total amount of 100 A in a hot water bath (oil phase) and separately
The components belonging to B are dissolved by heating (aqueous phase). Oil phase obtained
Add the aqueous phase to the mixture, stir to emulsify and cool to soften
A plaster was obtained.

[0018] Example 4 (milky lotion) Component Parts by weight Akebia extract 1 tristearate polyoxyethylene sorbitan 1 glyceryl monostearate 0.5 Squalane 6 glyceryl trioctanoate 2 cetyl octanoate 2 Stearyl alcohol 2-methoxy cinnamic acid Octyl 2, 1,3-butylene glycol 5 Glycerin 3 Nicotinamide 1 Preservative / antioxidant Appropriate amount Perfume Appropriate amount Purified water Appropriate amount ─────────────────── ─────────── Total amount 100 The above components were uniformly emulsified at 70 ° C to obtain an emulsion.

[0019] Example 5 (pack agent) Component Parts by weight Akebia extract 0.4 Dipropylene glycol 3 Polyethylene glycol 3 1,3-butylene glycol 1 Sodium 1 lactate 0.5 sodium lactate Glycerin 2-pyrrolidone-carboxylic acid 0.5 Polyvinyl alcohol 12 Ethanol 3 POE (10) tetradecyl ether 0.3 Preservative / antioxidant qs perfume qs Purified water qs ────────────── Total amount 100 The above components were uniformly dissolved to obtain a pack.

[0020] Example 6 (Lotion) Component Parts by weight Akebia extract 0.5 Glycerin 4 1,3-butylene glycol 4 Ethanol 7 POE (20) oleyl ether 0.5 Preservative-antioxidant q.s. incense Appropriate amount Purified water appropriate amount ────────────────────────────── Total amount 100

Glycerin and 1,3-butylene glycol were dissolved in purified water to obtain an aqueous solution. Separately, ethanol, Kimitsu extract and POE (20) oleyl ether were mixed. This mixture was added to the above aqueous solution, dissolved, and filtered to obtain a lotion.

[0022]

As described in detail above, the present invention provides a novel,
And, it can provide an effective arginase activity promoter, and in the form of a skin external preparation, has an excellent skin moisturizing effect of increasing the water content of the stratum corneum,
It is useful for improving rough skin and is highly safe.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁶ Identification code Agency reference number FI Technical display location A61K 7/48 A61K 7/48

Claims (2)

[Claims] An arginase activity promoter comprising an extract of Mokutsu as an active ingredient. 2. The arginase activity promoter according to claim 1, which is in the form of an external preparation for skin.