JP5144362B2 - Topical skin preparation - Google Patents

Description

  The present invention relates to a skin external preparation excellent in prevention and improvement of wrinkles, anti-inflammatory action and the like, characterized by containing an extract of Atemoya.

  In recent years, free radicals and active oxygen have been taken up as factors that oxidize biological components, and their adverse effects have become a problem. It is said that free radicals and active oxygen are generated in the body and decompose or crosslink biological tissues such as collagen, and oxidize fats and oils to form lipid peroxides that damage cells. Such a disorder is considered to cause skin wrinkles, a decrease in tension, and the like, and a method for suppressing the formation of lipid peroxide is known as one of the methods for preventing these problems. Conventionally, ascorbic acid, tocopherol, BHT (3,5-tert-butyl-4-hydroxytoluene), superoxide dismutase (SOD), and the like have been used as antioxidants.

  Further, it is known that wrinkles are caused by fiber reduction and degeneration of the dermal matrix such as collagen and elastin. In recent years, research has progressed and it has been pointed out that MMP is particularly involved as a factor inducing this change. Among the MMPs, collagenase and gelatinase are known as enzymes that degrade collagen, which is the main component of the dermal matrix of the skin, and its expression is greatly increased by irradiation with ultraviolet rays, which causes the decrease and degeneration of collagen by ultraviolet rays. It is considered to be one of the major factors such as the formation of wrinkles. Therefore, inhibiting these enzymes protects the collagen and protects the matrix that forms the fibers, and is important in preventing wrinkles. MMP activity inhibitors are known as one method for preventing such wrinkles. Conventionally, retinoic acid, α-hydroxy acid, retinol, and the like have been used as components for preventing wrinkles.

  There is skin inflammation caused by ultraviolet rays or the like as a cause of rough skin, and an external preparation for skin that subsides the inflammation and improves rough skin is desired. As one of the treatment methods, there is a method of suppressing the release of histamine which is a inflammatory substance that occurs during inflammation. Conventionally, treatments for rough skin have been performed using steroids, indomethacin or the like. However, the plant material Atemoya has not been studied for the lipid peroxidation inhibitory action, the MMP activity inhibitory action, and the histamine release inhibitory action, which are the indicators of wrinkle prevention and improvement and anti-inflammatory action described above.

  SOD used for the purpose of preventing or improving skin wrinkles or for antioxidant purposes is unstable, difficult to formulate, and tocopherol cannot be said to be sufficiently effective. Moreover, since BHT etc. which are synthetic compounds have a problem in safety | security and there exists a restriction | limiting in compounding quantity, it is not a chemical synthetic product but the natural raw material which is stable and has few side effects is desired. Similarly, it is preferable for prevention and improvement of wrinkles to have a safe and stable MMP activity inhibitory action. In addition, most of the conventional skin external materials that have the action of suppressing the release of histamine contain substances obtained by chemical synthesis such as steroids, and there is a risk of side effects. Therefore, a skin external preparation derived from a natural product having a high effect and an excellent effect is desired.

  From the above, a skin external preparation that is safe and excellent in stability, is excellent in prevention and improvement of wrinkles, and an anti-inflammatory action is desired.

  Under such circumstances, as a result of intensive studies, the present inventors have found that the extract of Atemoya has excellent lipid peroxide production inhibitory action, MMP activity inhibitory action and histamine release inhibitory action, and is also excellent in stability. I found. Furthermore, the skin external preparation containing the extract was found to be safe and stable, and excellent in prevention and improvement of wrinkles and an anti-inflammatory action, and the present invention was completed.

  Atemoya (scientific name: Annona atemoya) used in the present invention is a plant belonging to the genus Vanleci, and is an artificially improved variety of bream (Banleciaceae, scientific name: Annona squamosa) and cherimoya (Annona cherimola). It is a fruit tree. In fact, it is widely distributed for food use, but its use as a cosmetic has not been reported.

  The extract of Atemoya used in the present invention is extracted from a part of the plant body such as Atemoya leaves, stems, bark, flowers, fruits, roots or whole plants. Preferably, those extracted from fruits, pulp, pericarp, seeds and the like are good. The extraction method is not particularly limited, and for example, it may be a heat extraction or a room temperature extraction. Further, raw atemoya may be used, or a dried one may be used.

  Examples of the solvent to be extracted include water, lower alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), liquid polyhydric alcohols (1,3-butylene glycol, propylene glycol). , Glycerin, etc.), ketones (acetone, methyl ethyl ketone, etc.), acetonitrile, esters (ethyl acetate, butyl acetate, etc.), hydrocarbons (hexane, heptane, liquid paraffin, etc.), ethers (ethyl ether, tetrahydrofuran, propyl ether) Etc.). Preferred are polar solvents such as water, lower alcohols and liquid polyhydric alcohols, and particularly preferred are water, ethanol, 1,3-butylene glycol and propylene glycol. These solvents may be used alone or in combination of two or more.

  The extract may be used as it is, or may be used after concentration, dilution and filtration treatment, decolorization with activated carbon, deodorization treatment, or the like, if necessary. Further, the extracted solution may be subjected to a treatment such as concentration to dryness, spray drying, freeze drying, etc., and used as a dried product.

  In the external preparation for skin of the present invention, the above extract may be used as it is, and within the range not impairing the effect of the extract, oils and fats, waxes, hydrocarbons, fatty acids which are components used in the external preparation , Alcohols, esters, surfactants, metal soaps, pH adjusters, preservatives, fragrances, moisturizers, powders, UV absorbers, thickeners, pigments, antioxidants, whitening agents, chelating agents, etc. These components can be blended.

  The external preparation for skin of the present invention can be used for any of cosmetics, quasi drugs, and pharmaceuticals. Examples of the dosage form include skin lotions, creams, emulsions, gels, aerosols, essences, Examples include packs, cleaning agents, bath preparations, foundations, dusting powders, lipsticks, ointments, and poultices applied to the skin.

  The amount of the extract used in the present invention is 0.0001% by weight or more, preferably 0.001 to 10% by weight in terms of solid matter, based on the total amount of the external preparation for skin of the present invention. If it is less than 0.0001% by weight, a sufficient effect is hardly expected. When the blending amount exceeds 10% by weight, the effect is hardly recognized and it is uneconomical. In addition, the addition method may be added in advance or during the production, and may be appropriately selected in consideration of workability.

  The extract of Atemoya of the present invention had an excellent lipid peroxide production inhibitory action, MMP activity inhibitory action and histamine release inhibitory action, and was also excellent in stability. Furthermore, the external preparation for skin containing these extracts showed safe and excellent wrinkle prevention and improvement and anti-inflammatory action.

  Next, in order to describe the present invention in detail, examples of producing the extract used in the present invention, formulation examples and experimental examples of the present invention will be given as examples, but the present invention is not limited thereto. The part of the compounding amount shown in the prescription example means part by weight, and% means weight%.

Production Example 1 Hot water extract of Atemoya pericarp 400 g of purified water was added to 20 g of dried Atemoya pericarp, extracted at 95-100 ° C. for 2 hours, filtered, the filtrate concentrated, freeze-dried and 6.1 g of hot water extract was obtained.

Production Example 2 Hot Water Extract of Atemoya Fruit Add 400 g of purified water to 20 g of dried Atemoya fruit, extract at 95-100 ° C. for 2 hours, filter, concentrate the filtrate, freeze-dry, 10.4 g of hot water extract was obtained.

Production Example 3 Hot Water Extract of Atemoya Seeds 400 g of purified water was added to 20 g of dried products of Atemoya seeds, extracted at 95-100 ° C. for 2 hours, filtered, and the filtrate was concentrated and freeze-dried. 1.7 g of hot water extract was obtained.

Production Example 4 50% ethanol extract of Atemoya pericarp 400 g of 50 (v / v)% ethanol aqueous solution was added to 20 g of dried Atemoya pericarp, extracted at room temperature for 5 days, filtered, and the filtrate was concentrated and freeze-dried As a result, 7.1 g of 50% ethanol extract of Atemoya immature pericarp was obtained.

Production Example 5 Ethamo extract of Atemoya pericarp 400 g of ethanol was added to 20 g of dried Atemoya pericarp and extracted at room temperature for 5 days, followed by filtration. The filtrate was concentrated and freeze-dried to obtain an ethanol extract of Atemoya pericarp 0.2 g was obtained.

Formulation Example 1 Lotion Formulation Amount Hot water extract of Atemoya peel (Production Example 1) 0.1 part 2.1,3-butylene glycol 8.0
3. Glycerin 2.0
4). Xanthan gum 0.02
5). Citric acid 0.01
6). Sodium citrate 0.1
7). Ethanol 5.0
8). Methyl paraoxybenzoate 0.1
9. Polyoxyethylene hydrogenated castor oil (40E.O.) 0.1
10. Perfume
11. [Manufacturing method] Components 1 to 6 and 11 and components 7 to 10 are uniformly dissolved in purified water, and both are mixed and filtered to obtain a product.

Comparative Example 1 Conventional lotion In Formulation Example 1, the hot water extract of Atemoya peel (Production Example 1) was replaced with purified water to obtain a conventional lotion.

Formulation Example 2 Cream Formulation Amount 50 parts ethanol extract of Atemoya peel (Production Example 4) 0.05 part Squalane 5.5
3. Olive oil 3.0
4). Stearic acid 2.0
5). Beeswax 2.0
6). Octyldodecyl myristate 3.5
7). Polyoxyethylene cetyl ether (20E.O.) 3.0
8). Behenyl alcohol 1.5
9. Glycerol monostearate2.5
10. Fragrance 0.1
11. Methyl paraoxybenzoate 0.2
12 Ethyl paraoxybenzoate 0.05
13.1,3-Butylene glycol 8.5
14 [Manufacturing method] Components 2 to 9 are heated and dissolved and mixed with purified water to a total amount of 100, and kept at 70 ° C to obtain an oil phase. Ingredients 1 and 11 to 14 are dissolved by heating and mixed, and kept at 75 ° C. to form an aqueous phase. The aqueous phase is added to the oil phase to emulsify, and the mixture is cooled while stirring. The component 10 is added at 45 ° C., and further cooled to 30 ° C. to obtain a product.

Comparative Example 2 Conventional Cream In Formulation Example 2, a 50% ethanol extract of Atemoya peel (Production Example 4) was replaced with purified water to obtain a conventional cream.

Formulation Example 3 Emulsion Formulation Formulation 1. 1. Hot water extract of Atemoya fruit (Production Example 2) 0.001 part Squalane 5.0
3. Olive oil 5.0
4). Jojoba oil 5.0
5). Cetanol 1.5
6). Glycerol monostearate 2.0
7). Polyoxyethylene cetyl ether (20E.O.) 3.0
8). Polyoxyethylene sorbitan monooleate (20E.O.) 2.0
9. Fragrance 0.1
10. Propylene glycol 1.0
11. Glycerin 2.0
12 Methyl paraoxybenzoate 0.2
13. Make the total volume 100 with purified water
[Manufacturing method] Components 2 to 8 are dissolved by heating and mixed, and kept at 70 ° C to obtain an oil phase. Ingredients 1 and 10-13 are dissolved by heating and mixed, and kept at 75 ° C. to form an aqueous phase. The aqueous phase is added to the oil phase to emulsify, and the mixture is cooled while stirring. The component 9 is added at 45 ° C., and further cooled to 30 ° C. to obtain a product.

Formulation Example 4 Gel formulation Formulation amount 1. 1. 50 parts ethanol extract of Atemoya peel (Production Example 4) 1.0 part Ethanol 5.0
3. Methyl paraoxybenzoate 0.1
4). Polyoxyethylene hydrogenated castor oil (60 EO) 0.1
5). Perfume
6.1,3-Butylene glycol 5.0
7). Glycerin 5.0
8). Xanthan gum 0.1
9. Carboxyvinyl polymer 0.2
10. Potassium hydroxide 0.2
11. [Production method] Ingredients 2 to 5 and ingredients 1 and 6 to 11 are uniformly dissolved in purified water, and the two are mixed to obtain a product.

Formulation Example 5 Pack Formulation Formulation 1. 1. Hot water extract of Atemoya peel (Production Example 1) 0.1 part 50% ethanol extract of Atemoya peel (Production Example 4) 0.1
3. Polyvinyl alcohol 12.0
4). Ethanol 5.0
5.1,3-Butylene glycol 8.0
6). Methyl paraoxybenzoate 0.2
7). Polyoxyethylene hydrogenated castor oil (20 EO) 0.5
8). Citric acid 0.1
9. Sodium citrate 0.3
10. Perfume proper amount11. [Production Method] Components 1 to 11 are uniformly dissolved in purified water to make a total amount of 100 to obtain a product.

Formulation Example 6 Foundation Formulation Amount Ethanol extract of Atemoya peel (Production Example 5) 1.0 part Stearic acid 2.4
3. Polyoxyethylene sorbitan monostearate (20EO) 1.0
4). Polyoxyethylene cetyl ether (20E.O.) 2.0
5). Cetanol 1.0
6). Liquid lanolin 2.0
7). Liquid paraffin 3.0
8). Isopropyl myristate 6.5
9. Sodium carboxymethylcellulose 0.1
10. Bentonite 0.5
11. Propylene glycol 4.0
12 Triethanolamine 1.1
13. Methyl paraoxybenzoate 0.2
14 Titanium dioxide 8.0
15. Talc 4.0
16. Bengala 1.0
17. Yellow iron oxide 2.0
18. Perfume proper amount19. [Manufacturing method] Components 2 to 8 are heated and dissolved in purified water to a total amount of 100, and kept at 80 ° C to obtain an oil phase. Swell component 9 well in component 19, then add components 1 and 10-13 and mix uniformly. To this, components 14 to 17 pulverized and mixed with a pulverizer are added, and the mixture is stirred with a homomixer and kept at 75 ° C. to obtain an aqueous phase. The oil phase is added to this aqueous phase with stirring, cooled, component 18 is added at 45 ° C., and cooled to 30 ° C. with stirring to give a product.

Formulation Example 7 Bath preparation formulation 1. Atomoya seed hot water extract (Production Example 3) 5.0 parts Sodium bicarbonate 50.0
3. Yellow No. 202 (1) Appropriate amount 4. Perfume appropriate amount 5. [Manufacturing method] Ingredients 1 to 5 are mixed uniformly with sodium sulfate to make a product.

Formulation Example 8 Ointment Formulation Formulation 1. 1. Hot water extract of Atemoya peel (Production Example 1) 0.01 part 50% ethanol extract of Atemoya peel (Production Example 4) 0.5
3. Polyoxyethylene cetyl ether (30E.O.) 2.0
4). Glycerol monostearate 10.0
5). Liquid paraffin 5.0
6). Cetanol 6.0
7). Methyl paraoxybenzoate 0.1
8). Propylene glycol 10.0
9. [Production Method] Components 3 to 6 are heated and dissolved and mixed with purified water to a total amount of 100, and kept at 70 ° C. to obtain an oil phase. Ingredients 1, 2, and 7-9 are dissolved by heating and mixed, and kept at 75 ° C. to form an aqueous phase. The aqueous phase is added to the oil phase to emulsify, and the mixture is cooled to 30 ° C. with stirring to obtain a product.
Next, experimental examples will be given to explain the effects of the present invention in detail.

Experimental Example 1 Lipid peroxide production inhibitory action using human skin fibroblasts NB1RGB The lipid peroxide production inhibitory effect of fibroblasts by ultraviolet rays was measured by the TBA method.

  Samples 1, 4, and 5 were used as samples.

  Fibroblasts in the logarithmic growth phase were seeded at 3 × 10 4 cells in a φ60 mm dish, Eagles'MEM (containing 10% FBS) was added, and the cells were cultured at 37 ° C. under 5% CO 2 conditions. PBS added with a sample to a final concentration of 1.0 mg / mL after 6 days of culture was added, cultured at 37 ° C. under 5% CO 2 for 30 minutes, and then irradiated with UVA (6.5 J / cm 2). . After irradiation, the cells were detached from the dish, and the cells were ultrasonically disrupted. Then, 0.2 mL of the cell disruption solution, 0.2 mL of 8.1% SDS, 1.5 mL of 20% acetic acid (pH 3.5), 0.8% Sodium thiobarbiturate 1.5mL and distilled water 0.6mL were added and stirred well. Further, 0.01 mL of 400 mM BHT was added and reacted at 100 ° C. for 1 hour. After cooling, add 2 mL of 6.25% pyridine-containing n-butyl alcohol solution to 2 mL of the reaction solution, stir well, and centrifuge to measure the fluorescence intensity (excitation wavelength 515 nm, fluorescence wavelength 553 nm) of the n-butyl alcohol layer. And the amount of lipid peroxide. Furthermore, the thing which did not irradiate with an ultraviolet-ray was created, and the difference from what was irradiated with an ultraviolet-ray was made into the amount of lipid peroxide production. The lipid peroxide production inhibition rate was calculated from the ratio of the amount of lipid peroxide produced when the sample was added, with the amount of lipid peroxide produced when only the solvent was added instead of the sample being 100.

  These results are shown in Table 1. It was found that the extract of Atemoya peel has an extremely high lipid peroxide production inhibitory action.

Experimental Example 2 MMP activity inhibition test

  Production examples 1, 3, 4 and 5 were used as samples.

  A 10% SDS-PAGE gel (1 mm thickness) containing 0.6 mg / mL gelatin was prepared, and 0.015 mL of the culture supernatant of fibroblasts was electrophoresed under non-reducing conditions. Thereafter, the gel was washed twice with a solution of 2.5% Triton X-100 (manufactured by SIGMA) at room temperature for 30 minutes to remove SDS, and 200 mM sodium chloride, 5 mM calcium chloride, 0.01% brij-35 ( Incubation was carried out at 37 ° C. for 24 hours in 30 mM Tris-HCl buffer (pH 7.6) containing SIGMA. At this time, the sample was added to a buffer solution in which the gel was immersed. After completion of the reaction, the gel was stained with a solution of 0.2% Coomassie Brilliant Blue R (manufactured by SIGMA) and decolorized with a 5% methanol-7.5% acetic acid solution. Collagenase activity in a band near 72 kDa detected as a non-stained band on a blue gel was quantified with a densitometer (Atoden, Atodensitograph, AE-6905), and the activity inhibition rate was calculated. The inhibition rate was calculated from the ratio of the densitometer value when the sample was not added to 100 and the value when the sample was added.

  These results are shown in Table 2. It was confirmed that the extract of Atemoya has an extremely high inhibitory action on MMP activity.

Experimental Example 3 Histamine release inhibition test

  Samples 1, 4, and 5 were used as samples.

  A mast cell suspension collected from the abdominal cavity of Wister male rats (8 weeks old) was adjusted to 2 × 10 5 cells / mL, and 320 μL of an 8-fold dilution of rat serum containing IgE antibody was added to 650 μL of the suspension. Incubated for 30 minutes at ° C. Next, the sample was added to 1.0 or 0.3 mg / mL and incubated at 37 ° C. for 10 minutes. Furthermore, after adding 20 μg of ovalbumin and 30 μg of phosphatidylserine and incubating at 37 ° C. for 20 minutes for antigen stimulation, the mast cell suspension was cooled to 4 ° C. to stop the reaction. After cooling sufficiently, the mixture was centrifuged at 3,000 rpm for 5 minutes, and the supernatant containing the released histamine was collected. The supernatant histamine was quantified by adding hydrochloric acid in n-heptane to form histamine hydrochloride, and then measuring the fluorescence intensity by the o-phthalaldehyde method.

  These results are shown in Table 3. It was confirmed that the extract of Atemoya pericarp has a very high inhibitory action on histamine release.

Experimental example 4 Use test 1
One month use for 20 women (21 to 46 years old) using the lotion of Formulation Example 1, the cream of Formulation Example 2, the conventional lotion of Comparative Example 1 and the conventional cream of Comparative Example 2. A test was conducted. After use, the effect of improving skin wrinkles was determined by a questionnaire.

  The test results are shown in Table 4. The topical skin preparation containing the extract of Atemoya showed an excellent wrinkle-improving action. During the test period, there was no skin problem and there was no problem with safety. There was also no problem with the deterioration of the prescription ingredients.

Experiment 5 Use test 2
One month for 20 women (21 to 46 years old) suffering from rough skin using the lotion of Formulation Example 1, the cream of Formulation Example 2, the conventional lotion of Comparative Example 1 and the conventional cream of Comparative Example 2 A usage test was conducted. After use, the improvement effect of rough skin was judged by a questionnaire.

  The test results are shown in Table 5. The external preparation for skin containing the extract of Atemoya showed an excellent effect of improving rough skin. During the test period, there was no skin problem and there was no problem with safety. There was also no problem with the deterioration of the prescription ingredients.

  Examples 3 to 8 were also tested in the same manner, and showed excellent improving effects such as wrinkles and rough skin.

From the above, the extract of Atemoya of the present invention has excellent lipid peroxide production inhibitory action, MMP activity inhibitory action and histamine release inhibitory action, is highly safe, and can be used for cosmetics, foods, pharmaceuticals and the like. is there.

Claims (4)

An agent for preventing or improving wrinkles, which comprises an extract of Atemoya as an active ingredient . An anti-inflammatory agent comprising atemoya extract as an active ingredient . A rough skin improving agent comprising an extract of Atemoya as an active ingredient. A lipid peroxide production inhibitor comprising atemoya extract as an active ingredient.