JP3585154B2 - Anti-aging skin external preparation - Google Patents

Description

【００２０】
［真皮線維芽細胞代謝活性化作用］
ヒト由来真皮線維芽細胞を１ウェルあたり２．０×１０^４個となるように９６穴マイクロプレートに播種し、２４時間後に製造例１〜４に示したアガリクス茸子実体抽出物を含有する１．０容量％牛胎仔血清添加ダルベッコ最小必須培地にて、３７℃で４８時間培養した。次いで２−（４，５−ジメチル−２−チアゾリル）−３，５−ジフェニルテトラゾリウムブロミド（ＭＴＴ）を０．４ｍｇ／ｍｌ含有する前記培地に交換して３７℃で２時間培養し、テトラゾリウム環の開環により生じるフォルマザンを、２−プロパノールにて抽出し５５０ｎｍにおける吸光度により測定した。なお、１．０容量％牛胎仔血清添加ダルベッコ最小必須培地のみで培養した系を対照とし、５．０容量％牛胎仔血清添加ダルベッコ最小必須培地で培養した系を陽性対照とした。結果は対照における吸光度を１００．０％として表した活性化指数により表２に示した。
【００２１】
【表２】

【００２２】
その結果、表２に示したとおり、アガリクス茸子実体抽出物を添加して真皮線維芽細胞を培養することにより、活性化指数の上昇が認められ、有意な線維芽細胞代謝活性化が認められていた。特に、抽出溶媒として精製水及び８５％エタノールを用いた製造例１及び製造例２においては、０．０１重量％ときわめて低濃度で、有意な活性化指数の上昇が認められ、高い線維芽細胞賦活作用を有することが示された。
【００２３】
［紫外線による細胞傷害防御作用］
ヒト由来真皮線維芽細胞を１ウェルあたり２．０×１０^４個となるように９６穴マイクロプレートに播種し、２４時間後に製造例１〜４に示したアガリクス茸子実体抽出物を含有する５．０容量％牛胎仔血清添加ダルベッコ最小必須培地に交換し、３７℃で２４時間培養した。次いで培地をＨａｎｋｓ液に交換し、紫外線を０．５Ｊ／ｃｍ^２量照射した。再度、５．０容量％牛胎仔血清添加ダルベッコ最小必須培地に交換し、３７℃で２４時間培養した後、培地をニュートラルレッドを２０μｇ／ｍｌ含有する前記培地に交換して３７℃で２時間培養し、培地中に含まれるニュートラルレッドをリン酸緩衝生理食塩水で洗浄除去した。細胞内に取り込まれたニュートラルレッドを、０．１Ｎ塩酸含有３０％エタノール水溶液で抽出し、抽出液の５５０ｎｍの吸光度を測定した。ニュートラルレッドは、生細胞の細胞膜だけを透過し、リソゾームに沈着するので、生細胞だけを特異的に染色することができる。なお、アガリクス茸子実体の抽出物を添加せず、５．０容量％牛胎仔血清添加ダルベッコ最少必須培地のみで培養した系を対照とし、それぞれの紫外線照射後の細胞生存率を表３に示した。
【００２４】
【表３】

【００２５】
その結果、表３に示したとおり、アガリクス茸子実体抽出物を添加して真皮線維芽細胞を培養することにより、紫外線による真皮線維芽細胞の傷害に対し、有意な防御作用が認められていた。特に、抽出溶媒として精製水及び８５％エタノールを用いた製造例１及び製造例２においては、０．１重量％以下の低濃度で、６４％以上の細胞生存が認められ、紫外線による真皮線維芽細胞の傷害に対し、高い防御作用を有することが示された。
【００２６】
次に、先に示した製造例を用いて調製した実施例を示し、更に本発明について詳細に説明する。
【００２７】
［実施例１〜４］Ｏ／Ｗ乳化型美容液
表４に示したアガリクス茸子実体抽出物を用いて、下記の処方によりＯ／Ｗ乳化型美容液を調製した。
（処方）

製法：（１）〜（５）の油相成分を混合し７５℃に加熱して溶解，均一化する。一方（６）〜（８）の水相成分を混合，溶解して７５℃に加熱し、油相成分を添加して予備乳化する。（９）を添加した後ホモミキサーにて均一に乳化し、（１０）を加えてｐＨを調整する。冷却後４０℃にて（１１）〜（１３）を添加，混合，均一化する。
【００２８】
【表４】

【００２９】
前記実施例１〜実施例４を用いて、紫外線によるしわの発生に対する防止効果を評価した。なおアガリクス茸子実体抽出物を精製水に代替したものを比較例１とした。しわ発生防止効果は、ヘアレスマウス５匹を１群とし、各群について実施例及び比較例をそれぞれ１日１回背部に塗布し、１Ｊ／ｃｍ^２／週の長波長紫外線（ＵＶＡ）を５０週間照射し、ヘアレスマウスにおけるしわの発生状況を観察し、表５に示す判定基準に従って点数化して行った。この際、精製水のみを塗布した群を対照とした。結果は各群の平均値を算出し、ＵＶＡ照射日数との関係により表６に示した。
【００３０】
【表５】

【００３１】
【表６】

【００３２】
表６に示されるように、対照群においては、ＵＶＡ照射日数が４０週を越える頃には形成されたしわの深さが中程度にまで達し、５０週後には深いしわの発生が認められていた。これに対し、本発明の実施例塗布群では、いずれにおいても５０週後に微小ないし軽微なしわが認められた程度で、しわの発生は顕著に抑制されていた。一方比較例塗布群では、有意なしわの発生抑制或いは軽減は認められなかった。
【００３３】
続いて、本発明の実施例１〜実施例４及び比較例１について、抗炎症作用及び創傷治癒促進効果を評価した。人工的に炎症又は創傷を形成した１群５匹のマウスを用い、各群に実施例及び比較例をそれぞれ０．５ｇずつ１日２回７日間塗布し、７日目に炎症部位及び創傷部位の状態を観察した。抗炎症作用については「有効」，「やや有効」，「無効」、創傷治癒促進効果については「完全治癒」，「ほぼ治癒」，「治癒不完全」の３段階でそれぞれ評価し、各評価を得たマウスの数にて表７に示した。
【００３４】
【表７】

【００３５】
表７より明らかなように、抗炎症作用については、本発明の実施例塗布群ではいずれにおいても無効と評価されたマウスは見られず、３例以上のマウスにおいて有効な抗炎症作用が認められていた。また創傷治癒促進効果についても、本発明の実施例塗布群では創傷治癒の不完全なマウスはいずれにおいても認められておらず、３例以上のマウスで完全な治癒を認めていた。これに対し比較例１塗布群では、やや有効な抗炎症作用の認められたマウスが１例見られたが、残り４例では炎症の改善は全く認められなかった。また比較例１塗布群すべてにおいて、創傷治癒は不完全であった。
【００３６】
次に本発明の実施例１〜実施例４及び比較例１について、６ヶ月間の実使用試験を行った。パネラーとして、顕著なしわの発生等の皮膚症状を有する４０歳〜６０歳代の女性を用い、１群２０名とした。使用試験は、各群に実施例及び比較例のそれぞれをブラインドにて使用させて行った。使用試験前および使用試験終了後の皮膚の状態を観察し、しわの改善状況について、「改善」，「やや改善」，「変化なし」の３段階にて評価した。なお、しわの程度については写真撮影及びレプリカにより評価した。結果は、各評価を得たパネラー数にて表８に示した。
【００３７】
【表８】

【００３８】
表８に示されるように、しわの改善状況については、本発明の実施例使用群ではすべてにおいて改善傾向が認められていた。特に、実施例１及び実施例２使用群では、７５％及び８０％のパネラーで明確な改善を認めていた。これに対し、比較例使用群では、明確な改善を認めたパネラーは見られず、７５％のパネラーで症状の改善を認めなかった。
【００３９】
なお、本発明の実施例１〜実施例４については、上記使用試験期間中に含有成分の析出，分離，凝集，変臭，変色といった状態変化は全く見られなかった。また、各実施例使用群において、皮膚刺激性反応や皮膚感作性反応を示したパネラーは存在しなかった。
【００４０】
続いて本発明の他の実施例の処方を示す。
【００４１】
［実施例５］皮膚用ローション

製法：（１）〜（５）を混合し均一とする。
【００４２】
［実施例６］皮膚用乳剤

製法：（１）〜（６）の油相成分を混合，加熱して均一に溶解し、７０℃に保つ。一方、（７）〜（１０）の水相成分を混合，加熱して均一とし、７０℃とする。この水相成分に前記油相成分を攪拌しながら徐々に添加して乳化し、冷却した後４０℃にて（１１）を添加，混合する。
【００４３】
［実施例７］皮膚用ゲル剤

製法：（１）に（２）を均一に溶解した後、（３）に（４）を溶解して添加し、次いで（５）を加えて増粘させ、（６）を添加する。
【００４４】

製法：（１）〜（７）の油相成分を混合，溶解して７５℃に加熱する。一方、（８）〜（１０）の水相成分を混合，溶解して７５℃に加熱する。次いで、上記水相成分に油相成分を添加して予備乳化した後、ホモミキサーにて均一に乳化し、冷却後４０℃にて（１１），（１２）を添加，混合する。
【００４５】
［実施例９］水中油型乳剤性軟膏

製法：（１）〜（４）の油相成分を混合，溶解して均一とし、７５℃に加熱する。一方、（５）を（６）に溶解して７５℃に加熱し、これに前記油相成分を添加して乳化し、冷却後４０℃にて（７）を添加，混合する。
【００４６】
［実施例１０］化粧水

製法：（１）〜（４）を順次（５）に添加して均一に混合，溶解する。
【００４７】
［実施例１１］エモリエントクリーム（油中水型）

製法：（５），（６）を（９）の一部に溶解して５０℃とし、５０℃に加熱した（４）に攪拌しながら徐々に添加する。これをあらかじめ混合し７０℃に加熱溶解した（１）〜（３）に均一に分散し、これに（７），（８）を（９）の残部に溶解して７０℃に加熱したものを攪拌しながら添加し、ホモミキサーにて乳化する。冷却後、４０℃にて（１０），（１１）を添加，混合する。
【００４８】
［実施例１２］メイクアップベースクリーム

製法：（１）〜（４）の油相成分を混合し、７５℃に加熱して均一とする。一方（５）〜（７）の水相成分を混合し、７５℃に加熱，溶解して均一とし、これに（８）〜（１０）の顔料を添加し、ホモミキサーにて均一に分散させる。この水相成分に前記油相成分を添加し、ホモミキサーにて乳化した後冷却し、４０℃にて（１１）〜（１３）を添加，混合する。
【００４９】
［実施例１３］乳液状ファンデーション

製法：（１）〜（５）の油相成分を混合し、７５℃に加熱して均一とする。一方（６）〜（９）の水相成分を混合し、７５℃に加熱，溶解して均一とし、これに（１０）〜（１４）の顔料を添加しホモミキサーにて均一に分散させる。この水相成分に前記油相成分を添加し、ホモミキサーにて均一に乳化した後冷却し、４０℃にて（１５），（１６）を添加，混合する。
【００５０】

製法：（１）〜（６）の油相成分を混合，溶解して７５℃に加熱する。一方、（７）〜（９）の水相成分を混合，溶解して７５℃に加熱する。ついで、この水相成分に油相成分を添加して予備乳化した後、ホモミキサーにて均一に乳化して冷却し、４０℃にて（１０）を添加，混合する。
【００５１】
【発明の効果】
以上詳述したように、アガリクス茸子実体の抽出物は、真皮線維芽細胞賦活作用及び真皮線維芽細胞に対する紫外線による傷害を防御する作用を有し、これを含有する本発明の老化防止用皮膚外用剤は、シワ，シミの発生、皮膚弾性の低下といった皮膚老化症状の防止或いは改善に有効で、抗炎症作用，創傷治癒促進作用をも有し、さらに安定性，安全性も良好であった。[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention activates the metabolic activity of dermal fibroblasts and prevents the fibroblasts from being damaged by ultraviolet rays, thereby producing wrinkles and spots due to various stresses such as aging and ultraviolet rays, and reducing the elasticity of the skin. The present invention relates to an antiaging skin external preparation which is effective for preventing or improving skin aging symptoms and has an anti-inflammatory action and a wound healing promoting action.
[0002]
[Prior art]
For skin aging symptoms such as wrinkles, spots, and reduced skin elasticity caused by aging and external stresses such as ultraviolet rays, decreased function of fibroblasts in the skin dermis and reduction or degradation of matrix fibers are important factors. I have. Therefore, in order to obtain a skin external preparation for preventing aging that has the effect of preventing and improving skin aging, attempts have been made to search for and mix components having the activity of activating or promoting proliferation of fibroblasts.
[0003]
For example, loquat extract (JP-B-5-17206), α-hydroxyacetic acid (JP-A-5-112422), sterol ester of α-hydroxy acid (JP-A-8-104632), 6-benzylamino Purine (JP-A-7-233037), specific ribonuclease (JP-A-7-309778), L-lysyl-L-glycyl-L-histidine (JP-A-7-316192), milk-derived fibroblasts Growth factors (Japanese Patent Application Laid-Open No. Hei 8-119687), oxidized coenzyme A (Japanese Patent Application Laid-Open No. Hei 8-17561), and the like are disclosed.
[0004]
However, some of the above-mentioned components having a dermal fibroblast activating effect have insufficient action effects or poor stability, and are effective when contained in a skin external preparation base. Some required substantial amounts to be effective. In addition, those that have undesirable side effects or irritation, or that adversely affect the stability of the preparation, those that are difficult to mix with external preparations due to odor or color, and that have difficulty maintaining a certain level of action and quality There were many things.
[0005]
[Problems to be solved by the invention]
Therefore, in the present invention, a novel component having an excellent cell-activating effect for improving the metabolic activity of dermal fibroblasts is searched for, and by including it in a skin external preparation, skin damage and aging caused by external stress such as ultraviolet rays are investigated. The purpose of the present invention was to obtain an external preparation for preventing aging which is excellent in the effect of effectively preventing or improving the antioxidant.
[0006]
[Means for Solving the Problems]
In order to solve the above-mentioned problems, the present inventors screened a substance having an effective activating action using the metabolic activity promoting effect of dermal fibroblasts as an index. As a result, it was found that the extract of Agaricus blazei Murill fruit body had a high metabolic promoting effect on dermal fibroblasts and an effect of preventing damage to fibroblasts by ultraviolet rays. The extract of the fruit body of Agaricus blazei Murill has no adverse effects on the skin such as skin irritation and contact sensitization, and also activates dermal fibroblasts when incorporated in an external preparation for preventing aging. There was no inactivation of the action and the quality was stable.
[0007]
Agaricus blazei Murill is a kind of agaric genus of the basidiomycete agaricaceae agaricaceae, and is also called agaric agaric or himematsutake. Agaricus mushrooms are distributed in the southeast and south America of North America, and in the southeast of Brazil, are a type of mushroom that has been used by residents for a long time. Many reports have been made on the effectiveness of this Agaricus mushroom, especially on its antitumor activity. For example, a polysaccharide having an antitumor effect can be obtained by extracting the fruit body or mycelium of Agaricus mushroom with an aqueous solvent (JP-A-55-74797, JP-A-55-108292, JP-A-55-108292). JP-A 64-67194, JP-A-1-67195, JP-A-6-128164, etc., that a nucleic acid component having an antitumor effect can be obtained from the fruiting body of Agaricus mushroom (JP-A-1-66127). ), That a protein polysaccharide having antitumor activity is isolated from the hot water extraction residue of Agaricus mushroom (JP-A-2-78630), and that a glycoprotein having antitumor activity is isolated from 85% ethanol extraction residue of Agaricus mushroom What is obtained (Japanese Patent Application Laid-Open No. 6-9423), an ergosterol derivative having an antitumor activity present in the fruit body of Agaricus mushroom (Japanese Patent Application Laid-Open No. 1-246) 99 No.) and the like have been disclosed. In addition, as a physiological activity of Agaricus mushroom, a liver function improving agent containing Agaricus mushroom fruit body extract as an active ingredient (JP-A-2-124829) and an antiallergic effect of an Agaricus mushroom mycelium culture extract (JP-A-1-228480) JP-A No. 7-258107), and an effect of improving the immune function of an agaricus mushroom fruit body extract (JP-A-7-258107). The present inventors have newly discovered a remarkable fibroblast activating effect and an effect of preventing ultraviolet damage to dermal fibroblasts in a solvent extract of Agaricus mushroom fruit body, and completed the present invention. Was.
[0008]
BEST MODE FOR CARRYING OUT THE INVENTION
An embodiment of the present invention will be described.
[0009]
Agaricus blazei Murill used in the present invention is already widely known, and has been deposited with the National Institute of Bioscience and Human-Technology, National Institute of Advanced Industrial Science and Technology under the deposit number No. 4731 of the Institute for Bioscience and Biotechnology.
[0010]
When an extract is obtained from the fruit body of Agaricus mushroom, either a raw fruit body or a dried fruit body may be used. Examples of the extraction solvent for obtaining an extract of Agaricus mushroom fruit body include water, alcohols such as ethanol, methanol, isopropanol, isobutanol, n-hexanol, methylamyl alcohol, 2-ethylbutanol, and n-octyl alcohol; glycerin; Polyhydric alcohols such as ethylene glycol, ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, propylene glycol, propylene glycol monomethyl ether, propylene glycol monoethyl ether, triethylene glycol, 1,3-butylene glycol, hexylene glycol and derivatives thereof , Acetone, methyl ethyl ketone, methyl isobutyl ketone, ketones such as methyl-n-propyl ketone, ethyl acetate, isopropyl acetate, etc. Esters, ethyl ether, isopropyl ether, one or more mixed solvents selected from polar solvents such as ethers such as n- butyl ether can be preferably used. Further, a polar solvent to which an inorganic salt such as phosphate buffered saline is added, and a polar solvent to which a surfactant is added can be used, and there is no particular limitation. Among the above extraction solvents, one kind of solvent selected from ethanol, methanol, 1,3-butylene glycol and water or a mixed solvent of two or more kinds thereof, and a solvent obtained by adding an inorganic salt or a surfactant to these solvents Is preferably used.
[0011]
Further, extraction methods include a method of extracting by impregnation at room temperature, cooled or heated, a method of extraction using a distillation method such as steam distillation, and a method of directly extracting raw Agaricus mushroom to obtain an extract. For example, these methods may be used alone or in combination of two or more.
[0012]
The ratio of the Agaricus mushroom fruit body to the solvent at the time of extraction is not particularly limited. However, the solvent is 0.5 to 1000 times the weight of Agaricus mushroom fruit body 1, and the extraction rate and the efficiency are particularly preferably 0.1 to 1000 times. It is preferably 5 to 100 times by weight. The extraction temperature is conveniently in the range from room temperature to the boiling point of the solvent under normal pressure, and the extraction time varies depending on the extraction temperature and the like, but is preferably in the range of 2 hours to 2 weeks.
[0013]
The extract of Agaricus mushroom fruiting body thus obtained can be used as it is, but it is deodorized and decolorized within a range that does not lose the dermal fibroblast activating action and the cytotoxic protection action by ultraviolet rays. , Concentration or the like, or may be used as a fraction by column chromatography or the like. These extracts, purified products, and fractionated products can be dried by removing the solvent therefrom, and can be used as a solubilized form in a solvent such as alcohol or in the form of an emulsion to prevent aging. It can be added to an external preparation.
[0014]
The amount of the Agaricus mushroom fruit body extract to be added to the skin external preparation for preventing aging should be in the concentration range of 0.001 to 20% by weight in view of its effect, odor and color tone when added. desirable. If the compounding amount is less than 0.001% by weight, sufficient dermal fibroblast activating effect, cytotoxicity preventing effect by ultraviolet rays and anti-aging effect cannot be obtained, but it is not necessary to mix in a large amount, and 20% by weight. If it exceeds 50, the stability of the external preparation for aging prevention may be affected.
[0015]
In the present invention, an antiaging skin external preparation containing the above Agaricus mushroom fruit body extract may be provided. The antiaging skin external preparation may be in the form of a lotion, emulsion, cream, ointment or the like. be able to. Furthermore, lotions such as flexible lotion, astringent lotion, and lotion for washing, creams such as emollient cream, moisture cream, massage cream, cleansing cream, makeup cream, emollient emulsion, moisture emulsion, nourishing Emulsions such as emulsions and cleansing emulsions, packs such as jelly packs, peel-off packs, wash-off packs, powder packs, serums, and face wash can be provided as anti-aging cosmetics in various formulations. .
[0016]
In the present invention, other active ingredients such as other cell activators and whitening ingredients, humectants, anti-inflammatory agents, ultraviolet absorbers and the like can also be used in combination, and sunscreen cosmetics, skin protection cosmetics, whitening It can also be provided as a medicated cosmetic such as an agent or a quasi-drug.
[0017]
【Example】
First, a production example of an Agaricus mushroom fruit body extract used in Examples of the present invention will be described.
[0018]
[Production Examples 1-4]
Using the extraction solvents shown in Table 1, Agaricus mushroom fruit body extract was prepared. The dried Agaricus mushroom fruit body was pulverized, and a solvent 10 times the weight of the dried Agaricus mushroom fruit body was added. The resultant was immersed at room temperature for 3 days, and the filtrate was filtered to obtain an Agaricus mushroom fruit body extract.
[0019]
[Table 1]

[0020]
[Activation of dermal fibroblast metabolism]
Human-derived dermal fibroblasts were seeded in a 96-well microplate at 2.0 × 10 ⁴ cells per well, and 24 hours later contained 1 g of the Agaricus mushroom fruit body extract shown in Production Examples 1-4. The cells were cultured in Dulbecco's minimum essential medium supplemented with 0.0% by volume of fetal calf serum at 37 ° C. for 48 hours. Then, the medium was replaced with the above-mentioned medium containing 0.4 mg / ml of 2- (4,5-dimethyl-2-thiazolyl) -3,5-diphenyltetrazolium bromide (MTT), and cultured at 37 ° C. for 2 hours. Formazan generated by ring opening was extracted with 2-propanol and measured by absorbance at 550 nm. The system cultured in only Dulbecco's minimum essential medium containing 1.0% by volume of fetal bovine serum was used as a control, and the system cultured in Dulbecco's minimum essential medium containing 5.0% by volume of fetal bovine serum was used as a positive control. The results are shown in Table 2 by the activation index in which the absorbance in the control was expressed as 100.0%.
[0021]
[Table 2]

[0022]
As a result, as shown in Table 2, by adding the Agaricus mushroom fruit body extract and culturing the dermal fibroblasts, an increase in the activation index was observed, and significant activation of fibroblast metabolism was observed. I was In particular, in Production Examples 1 and 2 using purified water and 85% ethanol as the extraction solvent, a significant increase in the activation index was observed at a very low concentration of 0.01% by weight, and high fibroblasts were observed. It was shown to have an activating effect.
[0023]
[Protection of cell injury by ultraviolet rays]
Human-derived dermal fibroblasts were seeded at 2.0 × 10 ⁴ cells / well in a 96-well microplate, and 24 hours later, containing the Agaricus mushroom fruit body extract shown in Production Examples 1-4. The medium was replaced with Dulbecco's minimum essential medium supplemented with 0.0% by volume of fetal calf serum, and cultured at 37 ° C. for 24 hours. Next, the medium was replaced with Hanks' solution, and ultraviolet rays were irradiated at 0.5 J / cm ² . The medium was replaced again with Dulbecco's minimum essential medium supplemented with 5.0% by volume of fetal calf serum, and cultured at 37 ° C. for 24 hours. Then, the medium was replaced with the medium containing 20 μg / ml of neutral red and cultured at 37 ° C. for 2 hours. Then, neutral red contained in the medium was washed away with phosphate buffered saline. Neutral red incorporated into the cells was extracted with a 30% aqueous ethanol solution containing 0.1N hydrochloric acid, and the absorbance of the extract at 550 nm was measured. Neutral red penetrates only the cell membrane of living cells and deposits on lysosomes, so that only living cells can be specifically stained. The cell viability after irradiation with ultraviolet light is shown in Table 3 with the control cultured using only Dulbecco's minimum essential medium supplemented with 5.0% by volume of fetal calf serum without adding the extract of Agaricus mushroom fruit body. Was.
[0024]
[Table 3]

[0025]
As a result, as shown in Table 3, by adding the Agaricus mushroom fruit body extract and culturing the dermal fibroblasts, a significant protective effect against damage to the dermal fibroblasts by ultraviolet rays was recognized. . Particularly, in Production Examples 1 and 2 using purified water and 85% ethanol as the extraction solvent, at a low concentration of 0.1% by weight or less, cell survival of 64% or more was observed, and dermal fibroblasts due to ultraviolet rays were observed. It was shown to have a high protective effect against cell damage.
[0026]
Next, examples prepared using the above-mentioned production examples will be shown, and the present invention will be described in more detail.
[0027]
[Examples 1 to 4] O / W emulsified cosmetic serum Using the Agaricus mushroom fruit body extract shown in Table 4, O / W emulsified cosmetic serum was prepared according to the following formulation.
(Prescription)

Production method: The oil phase components (1) to (5) are mixed and heated to 75 ° C. to dissolve and homogenize. On the other hand, the aqueous phase components of (6) to (8) are mixed and dissolved, heated to 75 ° C., and the oil phase component is added and pre-emulsified. After the addition of (9), the mixture is uniformly emulsified with a homomixer, and the pH is adjusted by adding (10). After cooling, add (11)-(13) at 40 ° C., mix and homogenize.
[0028]
[Table 4]

[0029]
Using Examples 1 to 4, the effect of preventing the generation of wrinkles due to ultraviolet rays was evaluated. Comparative Example 1 was obtained by substituting the agaricus mushroom fruit body extract with purified water. The wrinkle prevention effect was obtained by applying five hairless mice to one group, applying the examples and the comparative examples to the back once a day for each group, and applying 1 J / cm ² / week long-wave ultraviolet light (UVA) for 50 weeks. Irradiation was carried out, and the occurrence of wrinkles in the hairless mouse was observed. At this time, a group to which only purified water was applied was used as a control. The results were calculated as the average value of each group, and are shown in Table 6 in relation to the number of days of UVA irradiation.
[0030]
[Table 5]

[0031]
[Table 6]

[0032]
As shown in Table 6, in the control group, the depth of wrinkles formed reached a moderate level when the number of UVA irradiation days exceeded 40 weeks, and deep wrinkles were observed after 50 weeks. Was. On the other hand, in each of the groups to which the examples of the present invention were applied, the occurrence of wrinkles was remarkably suppressed to the extent that slight or slight wrinkles were observed after 50 weeks in each case. On the other hand, in the comparative example application group, no significant suppression or reduction of wrinkles was observed.
[0033]
Subsequently, for Examples 1 to 4 and Comparative Example 1 of the present invention, the anti-inflammatory effect and the effect of promoting wound healing were evaluated. Using a group of 5 mice in which an inflammation or a wound was formed artificially, 0.5 g of each of Examples and Comparative Examples was applied to each group twice a day for 7 days. Was observed. The anti-inflammatory effect was evaluated in three stages: "effective", "slightly effective", "ineffective", and the effect of promoting wound healing in three stages: "complete healing", "almost healing", and "incomplete healing". The number of obtained mice is shown in Table 7.
[0034]
[Table 7]

[0035]
As is clear from Table 7, regarding the anti-inflammatory effect, none of the mice evaluated as ineffective in any of the groups to which the Examples of the present invention were applied, and an effective anti-inflammatory effect was observed in three or more mice. I was Regarding the effect of promoting wound healing, none of the mice with imperfect wound healing was observed in any of the groups to which Examples of the present invention were applied, and complete healing was observed in three or more mice. On the other hand, in the group to which Comparative Example 1 was applied, one mouse in which a slightly effective anti-inflammatory effect was observed was observed, but in the remaining four cases, no improvement in inflammation was observed at all. Further, in all the applied groups of Comparative Example 1, wound healing was incomplete.
[0036]
Next, a practical use test for 6 months was performed on Examples 1 to 4 and Comparative Example 1 of the present invention. As panelists, women in their 40s to 60s who had skin symptoms such as remarkable wrinkles were used, and a group of 20 women was used. The use test was performed by using each of the examples and the comparative examples blindly for each group. The skin condition was observed before the use test and after the use test, and the wrinkle improvement was evaluated in three stages of “improvement”, “slight improvement”, and “no change”. The degree of wrinkles was evaluated by photographing and replica. The results are shown in Table 8 by the number of panelists who obtained each evaluation.
[0037]
[Table 8]

[0038]
As shown in Table 8, the improvement of wrinkles was observed in all the groups using the examples of the present invention. In particular, in the use groups of Example 1 and Example 2, 75% and 80% of panelists showed a clear improvement. On the other hand, in the group using the comparative example, no panelists who recognized a clear improvement were observed, and 75% of the panelers did not improve the symptoms.
[0039]
In Examples 1 to 4 of the present invention, no state change such as precipitation, separation, agglomeration, discoloration, and discoloration of the components was observed at all during the use test period. In addition, in each group used in Examples, there was no paneler showing a skin irritating reaction or a skin sensitizing reaction.
[0040]
Subsequently, the prescription of another example of the present invention will be shown.
[0041]
[Example 5] Lotion for skin

Production method: (1) to (5) are mixed and made uniform.
[0042]
Example 6 Emulsion for skin

Production method: The oil phase components (1) to (6) are mixed and heated to uniformly dissolve, and kept at 70 ° C. On the other hand, the aqueous phase components (7) to (10) are mixed and heated to be uniform, and the temperature is set to 70 ° C. The oil phase component is gradually added to the aqueous phase component while stirring to emulsify, and after cooling, (11) is added and mixed at 40 ° C.
[0043]
[Example 7] Gel for skin

Production method: After (2) is uniformly dissolved in (1), (4) is dissolved and added in (3), then (5) is added to thicken, and (6) is added.
[0044]

Production method: The oil phase components (1) to (7) are mixed and dissolved and heated to 75 ° C. On the other hand, the aqueous phase components (8) to (10) are mixed and dissolved and heated to 75 ° C. Next, the oil phase component is added to the water phase component and pre-emulsified, then uniformly emulsified by a homomixer, and after cooling, (11) and (12) are added and mixed at 40 ° C.
[0045]
[Example 9] Oil-in-water emulsion ointment

Production method: The oil phase components (1) to (4) are mixed and dissolved to make uniform, and heated to 75 ° C. On the other hand, (5) is dissolved in (6), heated to 75 ° C., the oil phase component is added thereto to emulsify, and after cooling, (7) is added and mixed at 40 ° C.
[0046]
[Example 10] Lotion

Production method: (1) to (4) are sequentially added to (5) and uniformly mixed and dissolved.
[0047]
[Example 11] Emollient cream (water-in-oil type)

Production method: (5) and (6) are dissolved in a part of (9) to 50 ° C., and gradually added to (4) heated to 50 ° C. with stirring. This was previously mixed and uniformly dispersed in (1) to (3), which were heated and dissolved at 70 ° C., and (7) and (8) were dissolved in the remainder of (9) and heated to 70 ° C. Add while stirring and emulsify with a homomixer. After cooling, (10) and (11) are added and mixed at 40 ° C.
[0048]
[Example 12] Makeup base cream

Production method: The oil phase components of (1) to (4) are mixed and heated to 75 ° C. to be uniform. On the other hand, the aqueous phase components (5) to (7) are mixed, heated to 75 ° C. and dissolved to make uniform, and the pigments (8) to (10) are added thereto and uniformly dispersed by a homomixer. . The oil phase component is added to the aqueous phase component, emulsified by a homomixer, cooled, and (11) to (13) are added and mixed at 40 ° C.
[0049]
[Example 13] Emulsion foundation

Production method: The oil phase components (1) to (5) are mixed and heated to 75 ° C. to make uniform. On the other hand, the aqueous phase components (6) to (9) are mixed, heated and melted at 75 ° C. to make uniform, and the pigments (10) to (14) are added thereto and uniformly dispersed by a homomixer. The oil phase component is added to the aqueous phase component, uniformly emulsified by a homomixer, cooled, and (15) and (16) are added and mixed at 40 ° C.
[0050]

Production method: The oil phase components (1) to (6) are mixed and dissolved and heated to 75 ° C. On the other hand, the aqueous phase components (7) to (9) are mixed and dissolved, and heated to 75 ° C. Then, the oil phase component is added to the water phase component and pre-emulsified, then uniformly emulsified and cooled with a homomixer, and (10) is added and mixed at 40 ° C.
[0051]
【The invention's effect】
As described above in detail, the extract of Agaricus mushroom fruit body has an dermal fibroblast activating action and an action of protecting the dermal fibroblast from damage by ultraviolet rays, and the anti-aging skin of the present invention containing the same. The external preparation was effective in preventing or improving skin aging symptoms such as wrinkles and spots and a decrease in skin elasticity, and also had an anti-inflammatory action and a wound healing promoting action, and also had good stability and safety. .

Claims (2)

An external preparation for preventing aging, comprising an extract of the fruit body of Agaricus blazei Murill. An external preparation for preventing aging, which comprises a polar solvent extract of Agaricus blazei Murill fruit body.