200526265 (1) 九、發明說明 【發明所屬之技術領域】 本發明係關於對於隨著老化產生的皺紋，特別是曝曬 部位所產生的皺紋的改善效果優異，含有選自由具有使皮 膚在皮膚科學上及美容上保持健康狀態的效果之麥角固醇 及麥角固醇配糖體所成群之至少1種之皺紋改善劑。 【先前技術】 以人類爲首之所有生物的臟器從誕生成長之後，隨著 年齡的增加逐漸衰退，不久機能停止，而機能停止部分達 到一定程度上即到達死亡，此機能逐漸衰退狀態稱爲老 化。皮膚受到來自周圍環境的直接影響，雖然並非具有維 持身體內部環境之重要機能，無所有機能到達停止的情 況，但是其爲容易顯現出皺紋、老人斑、暗沈、鬆弛等老 化徵候之臟器，尤其以曝露在陽光下之曝曬部位特別明 §§ 〇 皮膚老化開始，則對於氧化應力等之剌激的防禦變 弱，成爲弄亂皮膚內部環境的原因，而使老化更惡化，特 別是曝曬部位經常曝露於紫外線等強氧化應力，老化的進 行會更顯著，如此的皮膚變化稱爲光老化，並未提出明確 的機轉，但複合各種要因所引起，結果成爲皮膚表面的皺 紋加深加粗等之美容上並不理想的狀態。 對於光老化進行結果所產生的皺紋具有改善效果的物 質，美國使用A酸（R e t i η 〇 i c A c i d )作爲處方箋，但因爲副200526265 (1) IX. Description of the invention [Technical field to which the invention belongs] The present invention relates to the effect of improving the wrinkles caused by aging, especially the wrinkles caused by the exposed part. A wrinkle improving agent containing at least one of ergosterol and ergosterol glycosides for maintaining a healthy state in cosmetics. [Previous technology] After the growth of the organs of all living creatures, including human beings, gradually declines with age, the function stops soon, and the part of the function stops reaching death to a certain extent. This state of gradual decline is called Ageing. The skin is directly affected by the surrounding environment. Although it does not have an important function to maintain the internal environment of the body, it does not have all the functions to stop, but it is an organ that easily shows signs of aging such as wrinkles, age spots, dullness, and relaxation. Especially when the exposed parts exposed to the sun are particularly clear §§ 〇 When skin aging begins, the defense against stimuli such as oxidative stress becomes weaker, which will cause the internal environment of the skin to be disturbed and worsen the aging, especially the exposed parts. Often exposed to strong oxidative stress such as ultraviolet rays, the progress of aging will be more significant. Such skin changes are called photoaging, and no clear mechanism is proposed. However, due to the combination of various factors, the wrinkles on the skin surface become deeper and thicker. Beauty is not ideal. For substances that have wrinkle improvement results as a result of photoaging, A-acid (R e t i η 〇 i c A c i d) is used as a prescription.
200526265 (2) 作用強且安全性亦有問題，日本至今並未認 「FRAGRANCE JOURNAL」、1 99 8 年 4 月 1 26卷、第4號、Ρ·75-77)，所以希望能開發$ 有充份效果之皺紋改善物質。 【發明內容】 本發明的目的爲提供對於老化、特別是曝 著的顯現出之皺紋的改善效果優異，且美容上 康皮膚效果優異之皺紋改善劑。 本發明者等鑑於上述情況進行精心硏究的 下述的皺紋改善劑改善因爲老化，特別是曝曬 現的皺紋，而且在美容上對於保持健康皮膚效 優異而達成本發明。 本發明爲關於添加選自由麥角固醇及麥角 所成群之至少1種爲特徵的皺紋改善劑。 本發明可提供對於隨著年齡增長特別是在 生的皺紋的改善效果優異，且美容上對於保持 果優異之皺紋改善劑。 [實施發明之最佳形態] 以下詳述本發明的實施形態。 本發明所使用的麥角固醇，作爲香菇及舞 所含有的機能性成份而被大家所熟知，據說具 疏鬆症的效果等，又市售產品由和光純藥工業 可（參考例如 ;日發行、第 ^全性高且具 曬部位之顯 對於保持健 結果，確認 部位明顯出 果及安全性 固醇配糖體 曝曬部位發 健康皮膚效 茸等菇類中 有預防骨質 公司、東京 -6- 200526265 (3) 化成工業公司、SIGMA-ALDRICH公司等所銷售，取得容 易。 麥角固醇配糖體可由麥角固醇經由如下述之規定方法 合成。 麥角固醇與乙醯溴- a -D-葡萄糖在用分子篩乾燥的氯 仿溶劑中，以四甲基脲及三氟甲基磺酸銀（silver triflate：) 作爲觸媒而反應，反應結束後藉由淸洗、濃縮、矽膠色譜 法進行純化、離析操作後，脫乙醯化即得到麥角固醇葡糖 苷。 本發明所使用的麥角固醇配糖體除了上述的葡萄糖以 外，可用半乳糖、甘露糖、葡糖胺、N -乙醯基葡糖胺、岩 藻糖、葡糖醛酸等單糖，及乳糖、麥芽糖等雙糖，或寡糖 合成，較佳爲可使用葡萄糖、半乳糖、木糖、乳糖、麥芽 糖合成，又此等糖之羥基、胺基、及羧基可被取代，亦可 使用例如日本國專利第3 3 4 3 0 3 6號所列舉之麥角固醇配糖 體衍生物。 本發明之選自由麥角固醇及麥角固醇配糖體所成·群之 至少1種，可單獨1種或組合2種以上，可爲組合麥角固 醇及麥角固醇配糖體者，亦可爲麥角固醇或麥角固醇配糖 體。再者配糖體存在具有α鍵結及/3鍵結之異構體，本發 明可使用任何一種，或此等的混合物，以具有；5鍵結者爲 佳。又本發明之選自由麥角固醇及麥角固醇配糖體所成群 之至少1種的添加量，以皺紋改善劑的總量爲基準，爲 0.00 1〜10.0質量％(下述只要無其它意義要旨的記載，％皆 200526265 (4) 表示質量％)爲佳，添加量只要位於此範圍，即可有效率及 充份得到本發明之目的效果，添加量較佳爲0 . 〇 5〜5.0 %， 更佳爲0.1〜2.0%。 本發明的皺紋改善劑可適合作爲皮膚化妝料及外用劑 之醫藥品及入浴劑等，可製成例如化妝水類、乳液類、面 霜類、面膜類等之劑型。 本發明之皺紋改善劑中在可達成本發明目的範圍內， 可適當添加上述以外的色素、香料、防腐劑、界面活性 劑、顏料、抗氧化劑等。 【實施方式】 實施例 以下基於實施例及比較例詳細說明本發明，但是本發 明的範圍並不限定於此等具體例。 [實施例1、2及比較例1 ] 經由下述的試驗方法’調查基劑單獨之試料，或添加 了麥角固醇或麥角固醇配糖體之試料，對於用於已光老化 的皮膚時之皺紋改善效果。 實施例中的麥角固醇配糖體係使用由下述製造方法所 得到者。 •麥角固醇（5.7g)及乙醯溴- a-D_葡萄糖（8.9g)，在經 由分子篩（5 g)乾燥的氯仿（1 〇 〇 m L)溶劑中，加入四甲基脲 (2.6 m L)及二—甲基擴酸銀（s i 1 v e r 11. i f 1 a t e )作爲觸媒，在遮 200526265 (5) 光、氮氣氣流下反應6小時’反應結束後過濾反應液 行洗淨、濃縮操作，使用矽膠色譜法（溶離溶劑：乙 -酯/已烷=1/4)進行純化、離析操作，將其脫乙醯化 . 1.2g的麥角固醇葡糖苷。 1. 動物實驗 試驗開始時使用1 〇週齡的無毛鼠1群1 〇隻。 2. 測量皺紋改善效果 • 2-1 ·光老化條件及測量方法 光老化係用UVA及UVB每天1次、週5次、照 週進行誘發，照射量以UVA爲 20J/cm2、25 J/cm2 J/cm2，UVB 爲 20J/cm2、30J/cm2、40 J/cm2 順序每 量，第3週之後照射最大量。 皺紋改善效果經由皺紋比分進行評估，皺紋比分 照 Bissett 寺之方法（Photochem Photobiol、46 : 3 7 8、1 9 8 7)進行計分，亦即，用肉眼總合評估皺紋的 ® 及深度，最高分爲3點，「可確認出粗且深的皴紋 3 ’ 「可確認出皴紋」爲2， 「無法確認出皺紋」爲 ~ 「觀察到正常的肌理」爲〇。 - 2-2·試料及實驗方法 50%(Wv)乙醇水溶液（基劑）中添加的麥角固醇 角固醇配糖體g周製試料（實施例1、2 )，又僅用基劑之 爲比較例1。 首先將0 · 1 m L的此等試料，於無毛鼠的背部皮fi 徑約2.5 c m)，以每天次、}週5次的頻率，自u v ，進 酸乙 得到 射8 、30 週增 爲遵 3 67- 粗細 」爲 或麥 試料 I (直 照射 -9 - 200526265 (6) 開始後第5週起至照射結束的第四週爲止進行塗抹，然後 塗抹結束後進行皺紋比分並計分，以塗抹基劑作爲對照組 比較。 3 .皺紋比分評估結果 群 皺紋比分値 實施例1 (添加麥角固醇之試料）塗抹群 2.45± 0.14 實施例2 (添加麥角固醇配糖體之試料）塗抹群 2.60± 0. 1 5 比較例1 (基劑試料）塗抹群 2.80± 0· 1 1200526265 (2) Strong function and safety issues, Japan has not yet recognized "FRAGRANCE JOURNAL", April 1, 1998, Vol. 26, No. 4, P · 75-77), so I hope to develop Fully effective wrinkle improving substance. [Summary of the Invention] The object of the present invention is to provide a wrinkle improving agent that is excellent in improving the effects of aging, particularly exposed wrinkles, and has an excellent effect on cosmetic skin. The present inventors have intensively studied the above-mentioned wrinkle-improving agent to improve wrinkles due to aging, particularly exposure, and have been found to be effective in maintaining healthy skin in terms of beauty. The present invention relates to the addition of a wrinkle improving agent characterized by adding at least one selected from the group consisting of ergosterol and ergot. The present invention can provide a wrinkle improving agent that is excellent in improving wrinkles with age, and is also excellent in cosmetically maintaining the fruit. [Best Mode for Carrying Out the Invention] Embodiments of the present invention will be described in detail below. The ergosterol used in the present invention is well-known as a functional ingredient contained in shiitake mushrooms and dances. It is said to have the effect of loosening, etc., and commercially available products are available from Wako Pure Chemical Industries (see, for example, daily release). 2. The most comprehensive and sun-exposed parts are shown to maintain healthy results. It is confirmed that the parts have obvious fruit and safety. Steroids and glucosides have healthy skin effect and mushrooms in the exposed parts. There are bone prevention companies, Tokyo-6- 200526265 (3) Available from Chemical Industries, SIGMA-ALDRICH, etc. It is easy to obtain. Ergosterol glycosides can be synthesized from ergosterol by the following method. Ergosterol and acetamidine-a- D-glucose was reacted with tetramethylurea and silver triflate: as catalysts in a chloroform solvent dried with a molecular sieve. After the reaction, the reaction was performed by washing, concentration, and silica gel chromatography. After purification and isolation operations, deacetylation yields ergosterol glucoside. In addition to the above-mentioned glucose, the ergosterol glycosides used in the present invention may be galactose, mannose, glucosamine, N -Synthesis of monosaccharides such as acetoglucosamine, fucose, glucuronic acid, and disaccharides such as lactose, maltose, or oligosaccharides, preferably glucose, galactose, xylose, lactose, and maltose In addition, the hydroxy, amine, and carboxyl groups of these sugars may be substituted, and for example, ergosterol glycoside derivatives listed in Japanese Patent No. 3 3 4 3 0 36 may be used. Free ergosterol and at least one species of ergosterol glycosides, which can be used alone or in combination of two or more. It can be a combination of ergosterol and ergosterol glycosides. It can be ergosterol or ergosterol glycosides. Furthermore, the glycoside has isomers with α-bond and / 3-bond. Any one or a mixture of these can be used in the present invention to have 5 bond is preferred. In addition, the amount of the at least one selected from the group consisting of ergosterol and ergosterol glycosides in the present invention is 0.00 1 based on the total amount of wrinkle improving agents. ~ 10.0% by mass (as long as there is no other meaning in the description below, the percentages are all 200526265 (4) represents mass%). Range, that is, the purpose and effect of the present invention can be effectively and fully obtained, and the added amount is preferably from 0.05 to 5.0%, more preferably from 0.1 to 2.0%. The wrinkle improving agent of the present invention can be suitably used as a skin cosmetic and external use Medicines and bathing agents such as agents can be formulated into, for example, lotions, lotions, creams, masks, etc. The wrinkle improving agent of the present invention may be appropriately added in addition to the above as long as it can achieve the purpose of the present invention. Pigments, flavors, preservatives, surfactants, pigments, antioxidants, etc. [Embodiments] Examples The present invention is described in detail below based on examples and comparative examples, but the scope of the present invention is not limited to these specific examples. [Examples 1, 2 and Comparative Example 1] According to the following test method 'Investigate base sample alone, or a sample added with ergosterol or ergosterol glycosides, Wrinkle improvement effect on skin. The ergosterol glycoside system in the examples used the one obtained by the following production method. • Ergosterol (5.7 g) and acetobromo-a-D-glucose (8.9 g), in a chloroform (1000 ml) solvent dried over a molecular sieve (5 g), add tetramethylurea ( 2.6 m L) and dimethyl silverate (si 1 ver 11. if 1 ate) as catalysts, and reacted under the light of 200526265 (5) for 6 hours under light and nitrogen gas flow. After the reaction, the reaction solution was filtered and washed. 2. Concentration operation, using silica gel chromatography (solvent: ethyl-ester / hexane = 1/4) for purification and isolation, and deacetylation. 1.2g of ergosterol glucoside. 1. Animal experiment At the beginning of the experiment, 10 groups of 10-week-old hairless mice were used. 2. Measure wrinkle improvement effect • 2-1 · Photoaging conditions and measuring methods Photoaging is induced by UVA and UVB once a day, 5 times a week, and weekly, and the irradiation dose is 20J / cm2, 25 J / cm2 J / cm2, UVB is 20J / cm2, 30J / cm2, 40 J / cm2 in order, the maximum amount after 3 weeks. The wrinkle improvement effect is evaluated by the wrinkle score. The wrinkle score is scored according to the method of Bissett Temple (Photochem Photobiol, 46: 3 7 8, 1 9 8 7). It is divided into 3 points, "Thick and deep wrinkles can be recognized 3 '," Wrinkles can be recognized "is 2," Wrinkles cannot be recognized "is ~" Normal texture observed "is 0. -2-2 · Test material and test method The test sample (Examples 1 and 2) prepared by adding ergosterol to sterol glycoside g week in 50% (Wv) ethanol aqueous solution (base), and only using the base This is Comparative Example 1. Firstly, apply these samples of 0 · 1 m L to the skin diameter of the hairless rat's back (approximately 2.5 cm in diameter), at a frequency of 5 times a day, 5 times a week, from the UV to the acid to get a shot of 8 and 30 weeks. In order to comply with 3 67-thickness "or wheat sample I (direct irradiation-9-200526265 (6) apply from the 5th week after the start to the fourth week after the end of the irradiation, and then wrinkle score and score after the end of the application, The smear base was used as a control group for comparison. 3. Results of wrinkle score evaluation group Wrinkle score 値 Example 1 (sample added with ergosterol) Smear group 2.45 ± 0.14 Example 2 (sample added with ergosterol glucoside ） Smudge group 2.60 ± 0. 1 5 Comparative Example 1 (base sample) Smudge group 2.80 ± 0 · 1 1
(値爲平均値±標準誤差） 實施例1與比較例1比較下顯示出有意義的低皺紋比 分値，又實施例2比實施例1差，但與比較例1比較，則 可看到皴紋比分減少的傾向，由此等的結果知道，麥角固 醇及麥角固醇配糖體顯示出對於光老化所誘發的皺紋有 效，具有改善因光老化所引起的皺紋之效果。 [實施例3及比較例2] 本實施例及比較例，遵照下述的調製法調製下述組成 的皮膚化妝水，以此作爲試料經由下述的操作，評估皺紋 改善效果。 5名具有眼尾紋之健康常人被檢驗者（女性，42〜56歲） 塗抹實施例或比較例的皮膚化妝水，用下述所示之方法進 -10- 200526265 (7) 行問卷調查關於眼尾部份皮膚（皺紋）的狀態。 請被檢驗者決定各試料分別用於左右那一邊，早上洗 完臉後，及晚上入浴後每天2次，2個月期間（6〇天）連續 在眼尾的皴紋部份（各試料塗抹於以眼尾中心之約4cm2、 2x 2cm)各塗抹約〇.2mL，接著最後塗抹結束後，請被檢 驗者回答關於左右眼尾部份皮膚（皺紋）的狀態。 W 1 ·皮膚化妝水的組成 原料成份 添加量（％) A成份 橄欖油 1〇.〇 肉豆蔻酸異丙酯 1.0 聚環氧乙烷（20)山梨糖醇酐月桂酸酯 0.5 丙二醇 1.0 甘油 2.0 B成份 對羥基苯甲酸甲酯 0.1 乙醇 7.0 純水 總量爲I 0 0時之餘量 c成份 麥角固醇 1.0(實施例3) 0 (比較例2) 200526265 (8) 2. 調製法 C成份的麥角固醇添加於B成份中均勻溶解後’添加 A成份混合攪拌分散，接著塡充至容器，使用時將內容物 均勻搖動振盪分散後使用。 3. 評估(値 is the average 値 ± standard error) Example 1 shows a significant low wrinkle ratio when compared with Comparative Example 1, and Example 2 is inferior to Example 1, but compared with Comparative Example 1, the wrinkles can be seen As a result, the score tends to decrease, and it is understood from these results that ergosterol and ergosterol glycosides are effective for wrinkles induced by photoaging and have an effect of improving wrinkles caused by photoaging. [Example 3 and Comparative Example 2] In this example and comparative example, a skin lotion of the following composition was prepared in accordance with the following preparation method, and a wrinkle improvement effect was evaluated by using the following operation as a sample. Five healthy subjects (women, 42 to 56 years old) with eye wrinkles apply the skin lotions of the examples or comparative examples, and use the method shown below -10- 200526265 (7) Questionnaire survey The condition of the skin (wrinkles) at the end of the eyes. The examinee is asked to decide which samples should be used for the left and right sides respectively, after washing the face in the morning, and twice a day after bathing at night, continuously on the creases at the end of the eyes for 2 months (60 days) (smear each sample) Apply about 0.2mL to each center (about 4cm2, 2x2cm) at the center of the eye end, and then after the last application, ask the examinee to answer the state of the skin (wrinkles) on the left and right eye ends. W 1 · Composition of skin lotion Ingredients Additives (%) A Ingredient Olive oil 10.0 Isopropyl myristate 1.0 Polyethylene oxide (20) Sorbitan laurate 0.5 Propylene glycol 1.0 Glycerin 2.0 B component methyl p-hydroxybenzoate 0.1 ethanol 7.0 remaining amount of pure water when I 0 0 c component ergosterol 1.0 (Example 3) 0 (Comparative Example 2) 200526265 (8) 2. Preparation method C Add the ergosterol to the B component and dissolve it evenly. 'Add the A component, mix, stir, and disperse. Then fill it into the container. When using, shake and distribute the contents evenly and shake and use. 3. Evaluation
以問卷結果爲基準，關於皮膚（皺紋）的狀態的各項 目，回答實施例3的皮膚化妝水有效於比較例2的人數表 示如下。 人數（名） 5 3 5 4 項目 皺紋變不明顯 皮膚變柔軟 皮膚變緊實 皮膚出現光澤 皮膚變亮 4 由本試驗結果知道實施例3的皮膚化妝水與比較例2 比較下明顯改善皺紋，而且就連因爲光老化而惡化之柔軟 性及皮膚的緊實亦改善，且未出現因爲本發明的皮膚化妝 水所產生的刺激及癢等皮膚的異常。 [實施例4] 下述組成的皮膚面霜依下述調製法調製，請事前所做 問卷調查中提出有眼尾紋的肌膚煩惱的2〇名健康常人（女 性’ 5 0〜5 6歲），使用1週以上後進行問卷調查。 -12 - 200526265 Ο) 1.皮膚面霜的組成 添加量（％) Α成份 •蜜鱲 2.0 、硬脂酸 5.0 硬脂醇 5.0 還原含水羊毛脂 2.0 角鯊烯 20.0 • 山梨糖醇酐硬脂酸酯 3.0 聚環氧乙烷（20)山梨糖醇酐硬脂酸酯 3.0 丙二醇 5.0 B成份 對羥基苯甲酸甲酯 〇.2 純水 總量爲1 〇 〇時之餘量 c成份 麥角固醇 1.0 2. 調製法 c成份的麥角固醇添加至B成份中，A、B成份分別 以8 0 °C加熱溶解後，混合持續攪拌，直到冷卻至3 0 °C調 製成各皮膚面霜。 3. 評估 對使用實施例4的皮膚面霜者實施問卷調查，其結果 如下述，又結果係進行問卷調查關於下述項目的皺紋狀 -13 - 200526265 (10) 態，列出各項目之與使用前比較而回答使用後者的人數。 項目 人數（名） 皺紋變不明顯 18 皺紋變細 17 皺紋數目減少 4 皺紋增加 0 由本試驗結果認爲實施例 4的皮膚面霜，與使用前比 較幾乎所有被檢驗者都覺得皺紋變不明顯，主要原因並非 皺紋數目減少而是皺紋變細，由此可知改善了因光老化所 產生的皺紋，且未出現因爲本發明的皮膚面霜所產生的刺 激及癢等皮膚的異常。 產業上的可利用性 可適用作爲皮膚化妝料及外用劑之醫藥品及入浴劑 等，可製成例如化妝水類、乳液類、面霜類、面膜類等之 劑型，非常適用於皮膚美容方面。 -14-Based on the results of the questionnaire, the number of people who answered the skin lotion of Example 3 with respect to the condition of the skin (wrinkles) is more effective than that of Comparative Example 2 is shown below. Number of people (name) 5 3 5 4 Items Wrinkles become inconspicuous Skin becomes soft Skin becomes firm Skin appears shiny Skin becomes bright 4 According to the results of this test, it is known that the skin lotion of Example 3 significantly improved wrinkles when compared with Comparative Example 2, and Even the softness and skin firmness deteriorated due to photoaging have been improved, and no skin abnormalities such as irritation and itching caused by the skin lotion of the present invention have occurred. [Example 4] The skin cream of the following composition was prepared according to the following preparation method, and 20 healthy people (women's 50 to 56 years old) who had troubled skin with eye wrinkles in the questionnaire survey beforehand were asked, Questionnaire survey after using for more than 1 week. -12-200526265 Ο) 1. Composition of skin cream (%) Α ingredients • honey 2.0, stearic acid 5.0 stearyl alcohol 5.0 reduced lanolin 2.0 squalene 20.0 • sorbitan stearate 3.0 polyethylene oxide (20) sorbitan stearate 3.0 propylene glycol 5.0 B component methyl parahydroxybenzoate 0.2 remaining amount when the total amount of pure water is 100 c component ergosterol 1.0 2. The ergot sterol of the c ingredient is added to the B ingredient. After the A and B ingredients are heated and dissolved at 80 ° C, they are mixed and stirred until they are cooled to 30 ° C to prepare skin creams. 3. Evaluation The questionnaire survey was carried out on those who used the skin cream of Example 4. The results are as follows. The results are a questionnaire about the wrinkles of the following items. 13-200526265 (10) State, list the use of each item. Compare the numbers while answering the former. Number of people (name) Wrinkles become inconspicuous 18 Wrinkles become fine 17 Wrinkles decrease 4 Wrinkles increase 0 According to the results of this test, the skin cream of Example 4 is considered to be less noticeable by almost all subjects than before use, mainly The reason is not that the number of wrinkles is reduced but that the wrinkles are becoming finer. From this, it is known that wrinkles due to photoaging are improved, and no skin abnormalities such as irritation and itching caused by the skin cream of the present invention have occurred. Industrial Applicability It can be used as skin cosmetics and external preparations for pharmaceuticals and bath preparations. It can be formulated into lotions, lotions, creams, masks, etc., and is very suitable for skin beauty. -14-