JP2000143527A - Hyaluronic acid production promoter and skin preparation for external use containing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a hyaluronic acid production promoter having an excellent hyaluronic acid production promoting action on fibroblast and improved stability and safety, suitable for external use and to obtain a skin preparation for external use useful for preventing and improving aging symptoms of skin, promoting treatment of wound and treating diseases followed by abnormal decomposition of hyaluronic acid by applying the promoter. SOLUTION: One or more kinds of extracts of Aloe arborescens Mill. with a polar solvent or a fraction obtained by successively fractionating an extract of Aloe arborescens Mill. with 50 vol.% ethanol by an adsorption resin chromatography made of a styrene polymer and a silica gel thin layer chromatography is used as it is or added to a carrier or a base to give a hyaluronic acid production promoter. One or more of hyaluronic acid production promoters are added to a base for external preparation to give the objective skin preparation for external use.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention has an effect of promoting the production of hyaluronic acid by skin fibroblasts, and is useful for preventing skin aging, promoting wound healing, and treating diseases associated with abnormal degradation of hyaluronic acid. The present invention relates to a hyaluronic acid production promoter and an external preparation for skin containing the same. More specifically, a hyaluronic acid production promoter containing one or more kinds of extracts of a polar solvent of Aloe arborescens Mill., Or a fraction thereof, and an external skin preparation containing the same Agent.

[0002]

2. Description of the Related Art Hyaluronic acid is a linear high molecular polysaccharide formed by alternately binding DN-acetylglucosamine and D-glucuronic acid, and constructs an extracellular matrix together with collagen, fibronectin, and proteoglycan. It has many functions such as cell retention, tissue lubricity retention, resistance to external forces such as mechanical injury, and prevention of bacterial infection. In particular, in the skin dermis, it is known that it is produced by fibroblasts and is involved in the elasticity maintenance of the skin and the wound healing process. It is also known that cell adhesion and cell migration are controlled via specific receptors.

[0003] Degradation of hyaluronic acid due to various external stresses such as aging and exposure to ultraviolet light, and a decrease in the hyaluronic acid-producing ability of fibroblasts, lead to the development of skin aging symptoms such as a decrease in skin elasticity and formation of wrinkles. It is thought to contribute to the delay of wound healing. In diseases such as rheumatoid arthritis, purulent arthritis, gouty arthritis, traumatic arthritis, osteoarthritis, and osteoarthritis, abnormal degradation of hyaluronic acid is observed. Therefore, in the prevention and improvement of such aging symptoms of the skin, promotion of wound healing, or treatment of diseases associated with abnormal degradation of hyaluronic acid, attempts have been made to supplement hyaluronic acid by external use or the like.

In the field of skin external preparations, many attempts have been made to apply hyaluronic acid or a salt thereof to an external preparation base (Japanese Patent Publication Nos. 60-19725 and 61-381).
68, 62-53484), in the treatment of the above-mentioned diseases accompanied by abnormal degradation of hyaluronic acid, joint injection therapy of hyaluronic acid is performed. However, regarding the application of skin external preparations containing hyaluronic acids, hyaluronic acid is a high molecular weight polysaccharide and is difficult to be percutaneously absorbed.
There has been a problem that hyaluronic acid is decomposed with time and the viscosity of the external preparation for skin decreases. In addition, joint injection therapy of hyaluronic acid requires long-term treatment,
There is a problem that it requires a doctor's prescription and is not suitable for daily treatment.

[0005] Therefore, in the external preparation for skin, the decrease in the viscosity of hyaluronic acid is suppressed to improve the stability of the preparation (Japanese Patent Publication No. 1-7043, 1-10485, etc.), or low-molecular-weight hyaluronic acid is used (Patent No. 2549119),
Or using a derivative of hyaluronic acid (Japanese Patent No. 2569)
No. 012, No. 2604930, No. 2648308
In addition to the attempts, many attempts have been disclosed to promote the production of hyaluronic acid in the living body or inhibit the activity of hyaluronidase to suppress the degradation of hyaluronic acid.

[0006] Such hyaluronic acid production promoters include:
Extracts of seaweeds such as Aosa genus, Aonori spp.
101871, 8-198741, 9-17603
6), plant extracts such as meso, reglo and lamiaceae (Japanese Patent Application Laid-Open Nos. 9-87163, 10-29922, 10-9)
5735), yeast extract (JP-A-8-163983),
A fraction having a molecular weight of 5,000 or less of bovine serum (JP-A-8-23)
9404) and the like. On the other hand, examples of the hyaluronidase inhibitor include those containing an extract of button and peonies (JP-A-1-128933) and those containing rosmarinic acid as an active ingredient (JP-A-9-6725).
1) and the like are disclosed.

However, the conventional hyaluronic acid production promoters and hyaluronidase inhibitors as described above are insufficient in any of hyaluronic acid production promotion effects or hyaluronidase inhibitory effects, quality, stability or safety. In fact, there has been a demand for a substance having a novel hyaluronic acid production promoting action.

[0008]

Accordingly, the present invention relates to a hyaluronic acid production promoter which has an excellent hyaluronic acid production promoting action on fibroblasts, is excellent in stability and safety, and is suitable for external use. The object of the present invention is to provide a skin external preparation which is useful for preventing and improving aging symptoms of the skin, promoting wound healing and treating diseases accompanied by abnormal degradation of hyaluronic acid.

[0009]

Means for Solving the Problems In order to solve the above problems, screening of a substance that enhances the hyaluronic acid-producing ability of fibroblasts without showing cytotoxicity was carried out, and the polarity of Aloe arborescens Mill. A high hyaluronic acid production-promoting activity was found in the extract by the solvent, and the present invention was completed by using the extract or a fraction thereof.

[0010] That is, in the present invention, one or more kinds of extracts of polar squid aloe ( Aloe arborescens Mill.) With a polar solvent, or squid aloe ( Aloe arborbor)
escens Mill.) in 50% by volume of ethanol, and then fractionated by styrene polymer adsorption resin chromatography and silica gel thin layer chromatography. This is used as a hyaluronic acid production promoter. Then, one or more of the above-mentioned hyaluronic acid production promoters are contained in an external preparation base to prepare a skin external preparation.

[0011]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Kidachi aloe ( Al) used as a starting material in obtaining the hyaluronic acid production promoter according to the present invention.
oe arborescens Mill.) is a plant belonging to the family Liliaceae native to South Africa, also called Ishairas, and widely cultivated in Japan. For extraction, each part and whole of leaves, stems, flowers, and roots can be used. These may be subjected to the extraction operation as they are, but considering the extraction efficiency, the shredded,
It is preferable to perform extraction after performing processing such as drying and pulverization. The extraction is performed by immersion in an extraction solvent. Stirring may be performed to increase the extraction efficiency, or homogenization may be performed in an extraction solvent. It is appropriate that the extraction temperature is set to a temperature of about 5 ° C. to the boiling point of the extraction solvent or lower. The extraction time varies depending on the type of the extraction solvent and the extraction temperature, but is from 4 hours to 14 hours.
It is appropriate to be on the order of days.

As an extraction solvent, in addition to water, methanol,
Lower alcohols such as ethanol, propanol and isopropanol; polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, dipropylene glycol and glycerin; ethers such as ethyl ether and propyl ether; esters such as ethyl acetate and butyl acetate And polar organic solvents such as ketones such as acetone and ethyl methyl ketone, and one or more of these can be selected and used. Further, physiological saline, phosphate buffer, phosphate buffered saline, or the like may be used. Among these solvents, it is preferable to use ethanol or a mixed solvent of ethanol and water, and it is particularly preferable to use 50% by volume of ethanol.

The extract of Kidachi aloe with the above-mentioned polar solvent can be used as it is as a hyaluronic acid production promoter, but the concentrated and dried extract may be redissolved in water or a polar solvent, or may have an effect of promoting hyaluronic acid production. May be used after performing purification treatments such as decolorization, deodorization, and desalting within a range that does not impair odor. For storage, it is preferable to freeze-dry after purification treatment and to dissolve in a solvent before use. In the present invention, a hyaluronic acid production promoter is obtained by extracting the extract of Kidachi aloe with a polar solvent or the above treated product as it is, or by including it in an aqueous carrier such as water or alcohol, or in a base such as emulsion, gel, cream or ointment. . Moreover, it can also be encapsulated in vesicles such as liposomes, microcapsules and the like.

[0014] In the present invention, Kidachiaroe (Al
(oe arborescens Mill.) can be used. That is, in the present invention, the extract is first fractionated by an adsorption resin chromatography made of styrene polymer with a mixed solvent of ethanol and water, and the fraction eluted with 30% by volume of ethanol is fractionated. The fraction that appeared as the spot with the highest polarity when developed with a mixed solvent of ethyl acetate-methanol-acetic acid having a volume ratio of 4: 1: 1 by layer chromatography was collected and used as such or as an aqueous carrier, emulsion, or gel. , Creams, ointments and the like, or contained in vesicles or microcapsules to form a hyaluronic acid production promoter. As the styrene polymer adsorption resin and silica gel, MCI gel (manufactured by Mitsubishi Chemical Corporation) and WAKO gel C-100
Commercial products such as (manufactured by Wako Pure Chemical Industries, Ltd.) can be used.

Further, in the present invention, one or more of the above-mentioned hyaluronic acid production promoters are contained in an external preparation base to provide a skin external preparation. The external preparation for skin according to the present invention can be provided in the form of lotions, emulsions, gels, creams, ointments and the like. Also, skin cosmetics such as lotions, emulsions, creams, packs, etc., makeup base lotions, makeup base creams, emulsion or creamy or ointment type foundations, makeup cosmetics such as eye color, cheek color, hand creams , Leg cream, body lotion and the like.

The external preparation for skin according to the present invention includes, in addition to the hyaluronic acid production promoter according to the present invention, general oils, surfactants, humectants, ultraviolet absorbers, pigments, fragrances, preservatives and the like. Raw materials for pharmaceuticals and cosmetics, and physiologically active ingredients such as active oxygen scavengers, anti-inflammatory agents, whitening agents, and skin cell activators. The amount of the hyaluronic acid production promoter according to the present invention in the external preparation for skin according to the present invention varies depending on the dosage form, but is 0.0001 to 10.0.
It is appropriate to set it to about% by weight.

[0017]

EXAMPLES Further, the features of the present invention will be described in detail with reference to examples.

Example 1 Hyaluronic acid production promoter 1 500 g of dry powder of leaves of Aloe arborescens Mill.
1 and left at 25 ° C. for 7 days. Thereafter, the plant powder was removed by filtration to obtain hyaluronic acid production promoter 1.

Example 2 Hyaluronic acid production promoter 2 The hyaluronic acid production promoter 1 of Example 1 was further added to MCI
Gel (manufactured by Mitsubishi Chemical Corporation) column chromatography, fractionated with ethanol-water mixed solvent, 30% by volume
The fraction eluted with ethanol was collected. Then, this fraction was subjected to thin layer chromatography on silica gel (WAKO gel C-100, manufactured by Wako Pure Chemical Industries, Ltd.) using a mixed solvent of ethyl acetate-methanol-acetic acid (volume ratio = 4: 1: 1) as a developing solvent. Then, the spot appearing on the most polar side was scraped off and dissolved in 100 ml of 25% ethanol by volume to obtain hyaluronic acid production promoter 2.

The effects of the hyaluronic acid production promoters of Examples 1 and 2 of the present invention on the hyaluronic acid production ability of normal human fibroblasts were examined. That is, normal human fibroblasts were seeded in a 96-well microplate at 2.0 × 10 ⁴ cells per well, and 1.0% by volume of cattle containing Example 1 or Example 2 at each test concentration. After culturing in fetal serum-supplemented Talbecco's modified basal medium (DMEM) for 5 days, the amount of hyaluronic acid in the culture supernatant was measured using Enzyme-l.
Inked immunosorbent assay
(ELISA). Simultaneously measure the number of fibroblasts, calculate the amount of hyaluronic acid production per cell,
Tables 1 and 2 show the hyaluronic acid production index in which the hyaluronic acid production amount of the control containing no sample was defined as 100.

[0021]

[Table 1]

As shown in Table 1, the hyaluronic acid production promoter 1 according to Example 1 of the present invention contained 1.56 to
The concentration of 12.5 μg / ml significantly promoted the production of hyaluronic acid in fibroblasts (risk 1%). The hyaluronic acid production promoter 1 at the above concentration increased the amount of hyaluronic acid production to about 3 to 4 times that of the control.

[0023]

[Table 2]

Further, as shown in Table 2, the hyaluronic acid production promoter 2 according to Example 2 of the present invention is 25 to 25.
Hyaluronic acid production of fibroblasts was significantly promoted in a concentration range of 1,000 μg / ml (hazard 1%), and 50 μg / ml
The amount of hyaluronic acid produced was 6 times higher than that of the control in ml and 7 times that of the control at 100 μg / ml. Example 1
No cytotoxicity was observed in any of the concentrations in the tested concentration range.

Further, another example of the hyaluronic acid production promoter according to the present invention will be described.

Example 3 Hyaluronic acid production promoter 3 10 g of olive oil and 30 g of a 10% by weight aqueous gelatin solution
The mixture is dispersed and emulsified at 40 ° C. with stirring to form an O / W emulsion. Add 4 to this emulsion in advance.
30 g of a 10% by weight aqueous solution of gum arabic heated to 0 ° C
Mix with stirring. In addition, the mixture was added at 40 ° C
Of warm water, and then add acetic acid in small portions to bring the pH to 4.0 to produce coacervate drops.
The system was cooled to 5 ° C. to coagulate the coacervate droplets, 1 ml of a 10% by weight aqueous glutaraldehyde solution was added, and the pH was adjusted to 9.0 with an aqueous sodium hydroxide solution. Raise the temperature. The resulting gelatin-gum arabic microcapsules were collected by centrifugation, olive oil was extracted with acetone, classified by sieving in acetone, and the acetone was evaporated and immersed in water to contain water. Next, hydrous gelatin-
The gum arabic microcapsules were immersed in the extract of a leaf of Kidachi aloe prepared in Example 1 above with a 50% by volume aqueous ethanol solution, and gelatin-gum arabic microcapsules containing the gum arabic microcapsules were prepared and collected by centrifugation. And hyaluronic acid production promoter 3.

[Example 4] Hyaluronic acid production promoter 4 The extract of the leaves of Kidachialoe prepared in Example 2 above was dissolved in 10 ml of 50% by volume ethanol, and this was dissolved in 10 ml of ethanol.
Add to 0 ml of purified water and dissolve. To this, 80 g of soybean lecithin was added, suspended at 65 ° C., then sonicated to prepare liposomes, and collected by centrifugation to obtain hyaluronic acid production promoter 4.

Next, the formulations of the examples of the external preparation for skin according to the present invention will be shown.

Example 5 Lotion for Skin (1) Ethanol 10.0 (% by weight) (2) Hydroxyethylcellulose 1.0 (3) Hyaluronic acid production promoter 1 0.1 (4) Methyl paraoxybenzoate 0.1 (5) Purified water 88.8 Production method: (1) to (4) are sequentially added to (5) and uniformly dissolved.

Example 6 Skin Emulsion (1) Stearic acid 0.20 (% by weight) (2) Cetanol 1.50 (3) Vaseline 3.00 (4) Liquid paraffin 7.00 (5) Polyoxy Ethylene (10E.O.) monooleic acid 1.50 ester (6) Tocopherol acetate 0.50 (7) Glycerin 5.00 (8) Methyl parahydroxybenzoate 0.10 (9) Potassium hydroxide 0.02 (10 ) Purified water 80.68 (11) Hyaluronic acid production accelerator 2 0.50 Production method: Mix and heat the oil phase components of (1) to (6), and uniformly dissolve them to 70 ° C. On the other hand, the aqueous phase components (7) to (10) are mixed and heated to be uniform, and the temperature is 70 ° C. The oil phase component is gradually added to this aqueous phase component while stirring to emulsify, and after cooling, (11) is added and mixed at 40 ° C.

Example 7 Skin Gel (1) Dipropylene glycol 10.0 (% by weight) (2) Carboxyvinyl polymer 0.5 (3) Potassium hydroxide 0.1 (4) Methyl paraoxybenzoate 0.1 (5) Purified water 88.8 (6) Hyaluronic acid production promoter 10.5 Production method: After (2) is uniformly dissolved in (5), (4) is dissolved in (1) Then, (3) is added to increase the viscosity, and (6) is added and mixed.

Example 8 Skin Cream (1) Beeswax 6.0 (% by weight) (2) Cetanol 5.0 (3) Reduced Lanolin 8.0 (4) Squalane 27.5 (5) Glyceryl fatty acid ester 4.0 (6) Lipophilic glyceryl monostearate 2.0 (7) Polyoxyethylene (20E.O.) sorbitan 5.0 Monolaurate (8) Propylene glycol 5.0 (9) Paraoxybenzoic acid Methyl 0.1 (10) Purified water 34.4 (11) Hyaluronic acid production promoter 3 3.0 Production method: Mix and dissolve oil phase components (1) to (7) to 75 ° C. On the other hand, the aqueous phase components (8) to (10)
Heat to ° C. Next, the oil phase component is added to the aqueous phase component, preliminarily emulsified, and then uniformly emulsified by a homomixer. After cooling, (11) is added at 40 ° C. and uniformly dispersed.

Example 9 Liposomal preparation for skin (1) Glycerin 2.0 (% by weight) (2) 1,3-butylene glycol 3.0 (3) Polyoxyethylene (25E.O.) oleyl ether 0 .2 (4) Ethanol 10.0 (5) Methyl paraoxybenzoate 0.1 (6) Fragrance 0.1 (7) Purified water 74.6 (8) Hyaluronic acid production promoter 4 10.0 Production method: (5 ) And (6) are dissolved in (4), added to (7) together with (1) to (3), mixed uniformly, and dispersed by adding (8).

Example 10 Oil-in-water emulsion ointment (1) White petrolatum 25.0 (% by weight) (2) Stearyl alcohol 25.0 (3) Glycerin 12.0 (4) Sodium lauryl sulfate 1.0 (5) Methyl paraoxybenzoate 0.1 (6) Hyaluronic acid production promoter 2 0.1 (7) Purified water 35.3 (8) Hyaluronic acid production promoter 3 1.5 Production method: (1) to (4) ) The oil phase components are mixed and heated to uniformly dissolve, and the temperature is adjusted to 75 ° C. On the other hand, the aqueous phase components (5) to (7) are mixed,
Heat to uniformity and bring to 75 ° C. The oil phase component is gradually added to the aqueous phase component while stirring to emulsify, and after cooling, (8) is added and dispersed at 40 ° C.

Example 11 Water-in-oil emollient cream (1) Liquid paraffin 30.00 (% by weight) (2) Microcrystalline wax 2.00 (3) Vaseline 5.00 (4) Diglyceryl dioleate Ester 5.00 (5) Sodium L-glutamate 1.60 (6) L-Serine 0.40 (7) Propylene glycol 3.00 (8) Methyl parahydroxybenzoate 0.10 (9) Hyaluronic acid production promoter 1 0.02 (10) Purified water 52.78 (11) Fragrance 0.10 Production method: (5) and (6) were dissolved in a part of (10) to 50 ° C, and preliminarily heated to 50 ° C ( Add slowly to 4) with stirring. This was previously mixed and uniformly dispersed in (1) to (3) heated and dissolved at 70 ° C., and then (7) to (9) were dissolved in the remainder of (10) and heated to 70 ° C. Add while stirring and emulsify with a homomixer. After cooling, at 50 ° C (1
Add 1) and mix.

Example 12 Makeup Base Cream (1) Stearic acid 12.00 (% by weight) (2) Cetanol 2.00 (3) Glyceryl tri-2-ethylhexanoate 2.50 (4) Self-emulsification Glyceryl monostearic acid 2.00 ester (5) Propylene glycol 10.00 (6) Potassium hydroxide 0.30 (7) Hyaluronic acid production promoter 2 0.05 (8) Purified water 69.55 (9) Oxidation Titanium 1.00 (10) Bengala 0.40 (11) Yellow iron oxide 0.10 (12) Fragrance 0.10 Manufacturing method: Mix and dissolve oil phase components (1) to (4) to 75 ° C . On the other hand, the aqueous phase components (5) to (8) are mixed and dissolved by heating, and the pigment components (9) to (11) are added thereto and uniformly dispersed by a homomixer to 75 ° C. Next, the oil phase component is added to the water phase component, and the mixture is uniformly emulsified by a homomixer. After cooling, (12) is added and mixed at 40 ° C.

Example 13 Emulsion Foundation (1) Stearic acid 2.00 (% by weight) (2) Squalane 5.00 (3) Octyldodecyl myristate 5.00 (4) Cetanol 1.00 (5) Decaglyceryl monoisopalmitate 9.00 (6) 1,3-butylenecricol 6.00 (7) Potassium hydroxide 0.08 (8) Methyl parahydroxybenzoate 0.10 (9) Purified water 53.42 (10) Titanium oxide 9.00 (11) Talc 7.40 (12) Bengala 0.50 (13) Yellow iron oxide 1.10 (14) Black iron oxide 0.10 (15) Fragrance 0.15 (16) Hyaluronic acid production promoter 2 0.15 Production method: Mix and dissolve oil phase components (1) to (5) to 75 ° C. On the other hand, the aqueous phase components (6) to (9) are mixed and dissolved by heating, and the pigment components (10) to (14) are added thereto and uniformly dispersed by a homomixer to 75 ° C. Next, the oil phase component is added to the aqueous phase component, and the mixture is uniformly emulsified by a homomixer. After cooling, (15) and (16) are added and mixed at 40 ° C.

Example 14 Hand Cream (1) Cetanol 4.00 (% by weight) (2) Vaseline 2.00 (3) Liquid paraffin 10.00 (4) Glyceryl monostearate 1.50 (5) Polyoxyethylene (60E.O.) glyceryl 2.50 Isostearate (6) Tocopherol acetate 0.25 (7) Glycerin 20.00 (8) Methyl parahydroxybenzoate 0.10 (9) Hyaluronic acid production promoter 1 0.04 (10) Purified water 59.61 Production method: Mix and dissolve the oil phase components (1) to (6) to 75 ° C. On the other hand, the aqueous phase components (7) to (10) were mixed and dissolved by heating.
5 ° C. Next, the oil phase component is added to the aqueous phase component, and the mixture is uniformly emulsified by a homomixer.

With respect to Examples 5 to 10 among the above Examples of the present invention, the protective effect against the formation of skin wrinkles by ultraviolet rays was evaluated. At that time, in each example, the hyaluronic acid production promoter 1 was impregnated with a 50% by volume aqueous ethanol solution, the hyaluronic acid production promoter 2 was impregnated with a 25 volume% aqueous ethanol solution, and the hyaluronic acid production promoter 3 was impregnated with a 50 volume% aqueous ethanol solution. Comparative Example 1 to Comparative Example 6 prepared by replacing the hyaluronic acid production promoter 4 with the liposomes prepared in the same manner as above without adding the extract of Kidachi aloe to the gelatin-gum arabic microcapsules thus prepared were also evaluated at the same time. Was done.

Protective effect against wrinkle formation by ultraviolet rays
Is performed for each group of 5 hairless mice
Example and Comparative Example were applied to the back once a day, respectively, and 1 J /
cm ²/ Week long-wave ultraviolet (UVA) irradiation for 50 weeks
And observe the formation of wrinkles on the hairless mouse skin
The score was evaluated according to the following criteria. This
At this time, a group to which only purified water was applied was used as a control. The result is
The average value of the group was calculated, and a table was displayed based on the relationship with the number of days of UVA irradiation.
3 is shown.

[Formation of wrinkles on skin] No wrinkle formation at all 0 point Slight formation of minute wrinkles 1 point Clear formation of slight wrinkles 2 points Medium wrinkle formation 3 points Deep wrinkles are recognized 4 points

[0042]

[Table 3]

As is clear from Table 3, in the control group, the depth of the wrinkles formed on the skin reached a medium level when the number of days of UVA irradiation exceeded 40 weeks, and became deep after 50 weeks. Formation was observed. On the other hand, in each of the groups to which the examples of the present invention were applied, the formation of fine or slight wrinkles was observed after 50 weeks, and the formation of wrinkles was remarkably suppressed. On the other hand, in the group to which the comparative example was applied, the degree of wrinkles formed in the group to which the comparative example 2 containing tocopherol acetate was applied was slightly reduced, but no significant suppression or reduction of wrinkles was observed.

Subsequently, the anti-inflammatory action and the wound healing promoting action of the above Examples 5 to 10 and Comparative Examples 1 to 6 were evaluated. Five mice, each of which had an artificial inflammation or wound on the back skin, were used as a group, and in each group, 0.5 g of each of Examples and Comparative Examples was applied to an inflamed site or a wound site twice a day for 7 days. . The state of each part was observed on the seventh day, and the anti-inflammatory effect was "effective", "slightly effective", "ineffective", and the wound healing promoting effect was "completely healed", "almost healed", "incomplete healing" The results are shown in Table 4 in terms of the number of mice that were evaluated in each of the three stages of "".

[0045]

[Table 4]

As can be seen from Table 4, no anti-inflammatory effect was observed in any of the groups to which the present invention was applied in any of the groups to which the present invention was applied, and no effective anti-inflammatory effect was observed in two or more mice. Was observed. Regarding the effect of promoting wound healing, none of the mice to which wound healing was incomplete was observed in any of the groups to which the Examples of the present invention were applied, and complete healing was observed in three or more mice. On the other hand, no effective anti-inflammatory action was observed in the group applied with the comparative example, except that a slightly effective anti-inflammatory action was observed in one case in the group applied with the comparative example 2. Also, in any application group,
Wound healing was incomplete.

Next, a practical use test for 6 months was performed on Examples 5 to 14 of the present invention. At this time, in Examples 11 to 14, the hyaluronic acid production promoter 1
Comparative Examples 7 to 10 were obtained by replacing the hyaluronic acid production promoter 2 with a 50% by volume aqueous ethanol solution and a 25% by volume aqueous ethanol solution in the same manner as described above. The actual use test was conducted in the age of 40 to 6 as a panelist, who exhibited remarkable skin aging symptoms such as wrinkles and decreased skin elasticity.
A group of 20 women in their 0s was used as a group, and each group was allowed to use the examples and comparative examples once each day in a blind. Observe the condition of the skin before and after the start of the use test, and evaluate the improvement of wrinkles and skin elasticity,
The evaluation was made in three stages of "slight improvement" and "no change". The degree of wrinkles was evaluated by taking photographs and collecting replicas, and the elasticity of the skin was evaluated by measuring with a cute meter. The results are shown in Table 5 by the number of panelists who obtained each evaluation.

[0048]

[Table 5]

As is evident from Table 5, all the panelists showed some improvement in wrinkles and skin elasticity in all the groups using the examples of the present invention. In particular, in the group using Example 5 to Example 10, clear improvement was recognized in wrinkles of 40% or more and skin elasticity of 65% or more of panelists. In contrast, in the group using the comparative example, none of the panelists showed a clear improvement in both wrinkles and skin elasticity, and there was no change in the skin condition with a paneler of 75% or more in both wrinkles and skin elasticity. I couldn't.

In Examples 1 to 4 of the present invention, the hyaluronic acid production promoting action was maintained almost unchanged for 6 months when stored at 10 ° C. or lower. Also, in Examples 5 to 14 of the present invention,
When stored at 25 ° C. for 6 months, no change in state was observed. Further, in the practical use test described above, in the group using the examples of the present invention, there were no panelists who recognized a skin irritating reaction or a skin sensitizing reaction, and the irritating sensation such as pain, warmth, burning, and tingling during use was observed. No panelists complained of discomfort.

[0051]

As described in detail above, the present invention has an excellent hyaluronic acid production promoting action on fibroblasts, and has good stability and safety, and is suitable for external use. Can be obtained and applied to prevent skin aging symptoms,
An external preparation for skin useful for improvement, promotion of wound healing, and treatment of diseases accompanied by abnormal degradation of hyaluronic acid could be provided.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI theme coat ゛ (reference) A61P 17/00 A61K 31/00 617 43/00 643C 643 F term (reference) 4C083 AA082 AA111 AA112 AB032 AB232 AB242 AB432 AC012 AC022 AC072 AC122 AC182 AC242 AC352 AC392 AC402 AC422 AC482 AC582 AC782 AD042 AD092 AD282 AD512 AD662 CC04 CC05 CC11 CC12 DD27 DD31 DD32 DD33 DD41 DD45 EE10 EE12 EE13 EE50 FF01 4C088 AB86 AC01 AC03 AC05 AC11 MA07 MA08 MA05 MA63 NA14 ZA89 ZB22 ZC80

Claims (4)

[Claims] [Claim 1] Aloe arborescens Mil
l.) A hyaluronic acid production promoter, which comprises one or more kinds of extracts with a polar solvent. 2. The hyaluronic acid production promoter according to claim 1, wherein the polar solvent is ethanol at 50% by volume. 3. Aloe arborescens Mil
l.) was extracted with a mixed solvent of ethanol and water by means of an adsorption resin chromatography made of styrene polymer, and a fraction eluted with 30% by volume of ethanol was collected. A hyaluronic acid production promoter comprising a fraction which appears as a spot on the highest polarity side when developed with a mixed solvent of ethyl acetate-methanol-acetic acid at a volume ratio of 4: 1: 1 by chromatography. 4. An external preparation for skin, comprising one or more selected from the hyaluronic acid production promoters according to claim 1. Description: