JP3933511B2 - Skin cosmetics and beauty food and drink - Google Patents

Description

【００５７】
表２に示すように、スターフルーツの果実抽出物を含有する試料溶液（試料溶液１〜３）は、グリセリン（保湿剤）を含有する試料溶液（試料溶液４）と同様に、精製水（試料溶液５）よりも水分残存率が高かった。このことから、スターフルーツの果実抽出物が保湿作用を有することが確認された。
【００５８】
〔試験例２〕スーパーオキサイド消去作用試験（ＮＢＴ法）
製造例１で得られた試料１〜３について、そのスーパーオキサイド消去作用を以下の方法により試験した。
３ｍＭキサンチン、３ｍＭ ＥＤＴＡ、１．５ｍｇ/ｍＬＢＳＡ溶液および０．７５ｍＭ ニトロブルーテトラゾリウム（ＮＢＴ）各０．１ｍＬと、０．０５Ｍ Ｎａ₂ＣＯ₃緩衝液（ｐＨ１０．２）２．４ｍＬとを試験管にとり、これに試料溶液０．１ｍＬを添加し、２５℃で１０分間放置した。次いでキサンチンオキシダーゼ溶液０．１ｍＬを加えて素早く攪拌し、２５℃で２０分間静置した。その後、６ｍＭ塩化銅０．１ｍＬを加えて反応を停止させ、波長５６０ｎｍにおける吸光度を測定した。以下、この吸光度を「試料溶液添加、酵素溶液添加時の吸光度」という。
【００５９】
また、同様の操作と吸光度の測定を、酵素溶液を添加せずに行った。このとき測定した吸光度を「試料溶液添加、酵素溶液無添加時の吸光度」という。
【００６０】
また、試料溶液を添加せずに蒸留水を添加した場合についても同様の測定を行った。このとき測定した吸光度を「試料溶液無添加、酵素溶液添加時の吸光度」という。
【００６１】
また、酵素溶液を添加せず、さらに試料溶液を添加せずに蒸留水を添加した場合についても同様の測定を行った。このとき測定した吸光度を「試料溶液無添加、酵素溶液無添加時の吸光度」という。
そして、次式によりスーパーオキサイド消去率を求めた。
【００６２】
消去率（％）＝｛１−（Ａ−Ｂ）／（Ｃ−Ｄ）｝×１００
【００６３】
上記式中、「Ａ」は「試料溶液添加、酵素溶液添加時の吸光度」、「Ｂ」は「試料溶液添加、酵素溶液無添加時の吸光度」、「Ｃ」は「試料溶液無添加、酵素溶液添加時の吸光度」、「Ｄ」は「試料溶液無添加、酵素溶液無添加時の吸光度」を表す。
【００６４】
試料濃度を段階的に減少させて上記消去率の測定を行い、スーパーオキサイドの消去率が５０％になる試料濃度（ｐｐｍ；μｇ/ｍＬ）を内挿法により求めた。その結果を表３に示す。
【００６５】
［表３］
試料 NO. 抽 出 物 ５０％消去試料濃度（ｐｐｍ）
１ 水抽出物 １９．０
２ ５０％エタノール抽出物 １６．９
３ エタノール抽出物 ２５．３
【００６６】
表３に示す結果から、スターフルーツの果実抽出物がスーパーオキサイド消去作用を有することが確認された。また、このスーパーオキサイド消去作用の程度は、抽出物の濃度によって調節できることが確認された。
【００６７】
〔試験例３〕ＤＰＰＨに対するラジカル消去作用試験
製造例１で得られた試料１〜３について、そのラジカル消去作用を下記の方法により非常に安定なラジカルであるＤＰＰＨを使用して試験した。
１．５×１０^-4Ｍ ＤＰＰＨエタノール溶液３ｍＬに試料溶液３ｍＬを加え、直ちに容器を密栓して振り混ぜ、３０分間静置した。その後、波長５２０ｎｍの吸光を測定した。コントロールとして、試料溶液の代わりに試料溶液を溶解した溶媒を用いて同様に操作し、波長５２０ｎｍの吸光度を測定した。また、ブランクとして、エタノールに試料溶液３ｍＬを加えたのち直ちに波長５２０ｎｍの吸光度を測定した。測定された各吸光度より、次式によりラジカル消去率（％）を算出した。
【００６８】
消去率（％）＝{１−（Ｂ−Ｃ）／Ａ}×１００
【００６９】
上記式中、「Ａ」は「コントロールの吸光度」、「Ｂ」は「試料溶液を添加した場合の吸光度」、「Ｃ」は「ブランクの吸光度」を表す。
【００７０】
試料濃度を段階的に減少させて上記消去率の測定を行い、ＤＰＰＨラジカルの消去率が５０％になる試料濃度（ｐｐｍ；μｇ/ｍL）を内挿法により求めた。その結果を表４に示す。
【００７１】
［表４］
試料 NO. 抽 出 物 ５０％消去試料濃度（ｐｐｍ）
１ 水抽出物 ２５．０
２ ５０％エタノール抽出物 ２６．０
３ エタノール抽出物 ２８．５
【００７２】
表４に示す結果から、スターフルーツの果実抽出物がラジカル消去作用を有することが確認された。また、このラジカル消去作用の程度は、抽出物の濃度によって調節できることが確認された。
【００７３】
〔試験例４〕一重項酸素消去作用試験
製造例１で得られた試料１〜３について、その一重項酸素消去作用を以下の方法により試験した。
透明ガラス瓶（１０ｍＬ容）中で２％赤血球懸濁液５ｍＬ、試料を所定濃度で含むｐＨ７．４の等張リン酸緩衝液５ｍＬ、および光増感剤（１０ｍＭヘマトポルフイリン−２０ｍＭ水酸化ナトリウム溶液）０．０１ｍＬを混合した。得られた溶液をメリーゴーランド上、７．５Ｗハロゲンランプで３５分間均一に照射して一重項酸素（^１Ｏ₂）を発生させ、赤血球の溶血を生じさせた。この反応溶液１ｍＬを採取し、等張リン酸緩衝液２ｍＬを加えて混合後、４℃、３０００ｒｐｍで５分間遠心分離を行った。次いで上清を採取し、波長５４０ｎｍの吸光度を測定した。別に、赤血球を一部溶血させた上記反応溶液１ｍＬをとり、これに蒸留水２ｍＬを加えて完全に溶血させたものをコントロールとし、同様に吸光度測定を行った。測定された吸光度より次式により一重項酸素消去率を求めた。
【００７４】
一重項酸素消去率（％）＝（１−Ｂ／Ａ）×１００
【００７５】
上記式中、「Ａ」は「コントロールの吸光度」、「Ｂ」は「反応溶液上清の吸光度」を表す。
【００７６】
試料濃度を段階的に減少させて上記消去率の測定を行い、一重項酸素の消去率が５０％になる試料濃度（ｐｐｍ；μｇ／ｍＬ）を内挿法により求めた。その結果を表５に示す。
【００７７】
［表５］
試料 NO. 抽 出 物 ５０％消去試料濃度（ｐｐｍ）
１ 水抽出物 １４７．３
２ ５０％エタノール抽出物 １２２．５
３ エタノール抽出物 １１８．４
【００７８】
表５に示す結果から、スターフルーツの果実抽出物が一重項酸素消去作用を有することが確認された。また、この一重項酸素消去作用の程度は、抽出物の濃度によって調節できることが確認された。
【００７９】
〔試験例５〕ヒアルロン酸産生促進作用試験
製造例１で得られた試料１〜３について、そのヒアルロン酸産生促進作用を以下の方法により試験した。
ヒト正常新生児線維芽細胞（NB1RGB）１×１０^６個を、７５ｃｍ²フラスコを用いて１０％ＦＢＳを含むα−ＭＥＭ培地（GIBCO）（ｐＨ７.２）で３７℃、５％ＣＯ_２−９５％ａｉｒの下にて７日間培養した。トリプシン処理により細胞を集め、１％ＦＢＳを含むα−ＭＥＭ培地を用いて２．２×１０⁴個／ｍＬに調整し９６穴のマイクロプレートに１００μＬづつ播種し、３７℃、５％ＣＯ_２−９５％ａｉｒの下で一晩培養した。翌日、試料（試料濃度：５０ｐｐｍ又は２００ｐｐｍ）（ｐｐｍ＝μｇ/ｍＬ）を溶解した１％ＦＢＳを含むα−ＭＥＭ培地を各ｗｅｌｌに１００μＬずつ添加し、３７℃、５％ＣＯ_２−９５％ａｉｒの下で３日間培養した。
【００８０】
培養上清１０μＬを９０μＬのＰＢＳ(−)で１０倍希釈し、その５０μＬを、あらかじめヒアルロン酸でコーティングしておいたＥＬＩＳＡプレートに添加して各種抗体を用いてＥＬＩＳＡを行った。ヒアルロン酸の定量は検量線を用いて行った。ヒアルロン酸産生促進率は、試料無添加時の値を１００％として求めた。その結果を表６に示す。
【００８１】

【００８２】
表６に示される結果より、スターフルーツの果実抽出物がヒアルロン酸産生促進作用を有することが確認された。また、このヒアルロン酸産生促進作用の程度は、抽出物の濃度によって調節できることが確認された。
【００８３】
〔試験例６〕エラスターゼ阻害作用試験
製造例１で得られた試料１〜３について、そのエラスターゼ阻害作用を以下の方法により試験した。
９６ウェルプレートを用意し、１穴に対して試料溶液（溶媒：ＤＭＳＯ＋水）５０μＬおよびエラスターゼ溶液５０μＬを添加し、さらに基質溶液１００μＬを添加し混合した。２５℃で１５分間反応させた後、波長４１５ｎｍの吸光度を測定した。上記と同様の酵素反応と吸光度測定を、試料溶液の代わりに試料溶液と等量の溶媒のみを添加して行った。さらに、それぞれの場合について、エラスターゼ溶液の代わりに緩衝液を添加して同じ操作と測定を行った。
【００８４】
なお、エラスターゼ溶液はシグマ社・エラスターゼTypeIII ５ｍｇをｐＨ８の０．２ｍｏｌ／Ｌ トリス塩酸緩衝液１ｍＬに溶解し使用時に２５０倍に希釈したものを使用した。基質溶液として、シグマ社のN−SUCCINYL−ALA−ALA−ALA p-NITROANILIDEをＤＭＳＯに溶解した濃度４５．１４ｍｇ／ｍＬの溶液を上記トリス塩酸緩衝液で１００倍に希釈して使用した。
測定結果より、次式に基づきエラスターゼ阻害率（％）を求めた。
【００８５】
エラスターゼ阻害率（％）＝〔１−（Ａ−Ｂ）／（Ｃ−Ｄ）〕×１００
【００８６】
上記式中、「Ａ」は試料溶液添加・酵素添加時の吸光度、「Ｂ」は試料溶液添加・酵素無添加時の吸光度、「Ｃ」は試料無添加・酵素添加時の吸光度、「Ｄ」は試料無添加，酵素無添加時の吸光度を表す。
【００８７】
試料濃度を段階的に減少させて上記阻害率の測定を行い、エラスターゼの活性を５０％阻害する試料溶液濃度（ｐｐｍ；μｇ／ｍＬ）を内挿法により求めた。
試験の結果を表７に示す。
【００８８】
[表７]
試料 NO. 抽 出 物 ５０％阻害試料濃度（ｐｐｍ）
１ 水抽出物 ２００
２ ５０％エタノール抽出物 １８２
３ エタノール抽出物 １７５
【００８９】
表７に示すように、スターフルーツの果実抽出物がエラスターゼ阻害作用を有することが確認された。また、このエラスターゼ阻害作用程度は、抽出物の濃度によって調節できることが確認された。
【００９０】
〔試験例７〕コラゲナーゼ阻害作用の試験
製造例１で得られた試料１〜３について、そのコラゲナーゼ阻害作用を以下の方法により試験した。
試料溶液（溶媒：２０ｍｍｏｌ／Ｌ 塩化カルシウム含有０．１ｍｏｌ／Ｌ トリス塩酸緩衝液（ｐＨ７．１）：以下の緩衝液においても同じ）５０μＬ、コラゲナーゼ溶液５０μＬおよび基質溶液４００μＬを混合し、３７℃で３０分間インキュベーションした。次いで２５ｍｍｏｌ／Ｌ クエン酸溶液１ｍＬで反応を停止し、酢酸エチル５ｍＬで抽出した。得られた抽出液について、波長３２０ｎｍの吸光度（対照液：酢酸エチル）を測定した。このとき測定した吸光度を「酵素添加, 試料溶液添加時の吸光度」という。なお、コラゲナーゼ溶液はシグマ社のコラゲナーゼＴｙｐｅＩＶ ５ｍｇを緩衝液１ｍＬに溶解させ、使用時に５０倍に希釈したものを使用した。基質溶液には、上記緩衝液にＢＡＣＨＥＭ Ｆｅｎｉｃｈｅｍｉｋａｌｉｅｎ ＡＧ社Ｐｚ−ペプチドを濃度が０．５ｍｏｌ／Ｌになるように溶解して使用した。
【００９１】
また、上記と同様の酵素反応と吸光度測定を、試料溶液の代わりに試料溶液と等量の緩衝液を添加して行った。このとき測定した吸光度を「酵素添加, 試料溶液無添加時の吸光度」という。
【００９２】
また、上記と同様の酵素反応と吸光度測定を、コラゲナーゼ溶液の代わりに緩衝液を添加して行った。このとき測定した吸光度を「酵素無添加, 試料溶液添加時の吸光度」という。
【００９３】
また、上記と同様の酵素反応と吸光度測定を、試料溶液の代わりに試料溶液と等量の緩衝液を添加するとともに、コラゲナーゼ溶液の代わりに緩衝液を添加して行った。このとき測定した吸光度を「酵素無添加, 試料溶液無添加時の吸光度」という。
そして、次式によりコラゲナーゼ阻害率（％）を算出した。
【００９４】
コラゲナーゼ阻害率（％）＝{１−（Ａ−Ｂ）／（Ｃ−Ｄ）}×１００
【００９５】
上記式中、「Ａ」は「酵素添加, 試料溶液添加時の吸光度」、「Ｂ」は「酵素無添加, 試料溶液添加時の吸光度」、「Ｃ」は「酵素添加, 試料溶液無添加時の吸光度」、「Ｄ」は「酵素無添加, 試料溶液無添加時の吸光度」を表す。
【００９６】
試料濃度を段階的に減少させて上記阻害率の測定を行い、コラゲナーゼの活性を５０％阻害する試料濃度(ｐｐｍ；μｇ／ｍＬ)を内挿法により求めた。その結果を表８に示す。
【００９７】
［表８］
試料 NO. 抽 出 物 ５０％阻害試料濃度（ｐｐｍ）
１ 水抽出物 ６８．０
２ ５０％エタノール抽出物 ５３．２
３ エタノール抽出物 ５０．１
【００９８】
表８に示す結果から、スターフルーツの果実抽出物がコラゲナーゼ阻害作用を有することが確認された。また、このコラゲナーゼ阻害作用の程度は、抽出物の濃度によって調節できることが確認された。
【００９９】
〔試験例８〕肌荒れ改善作用（皮膚の老化予防・改善作用）の試験
製造例１で得られたスターフルーツの果実からの５０％エタノール抽出物（試料２）を配合した乳液（以下「実施例乳液」という。）を常法に従って調製した。実施例乳液の組成を以下に示す。
【０１００】
スターフルーツ果実抽出物（製造例１の試料２） ０．１ｇ
セチルアルコール ０．５ｇ
ミツロウ ２．０ｇ
1,3-ブチレングリコール ３．０ｇ
オレイン酸ポリオキシエチレンソルビタン（20E.0） ０．５ｇ
ヒアルロン酸 ０．１ｇ
クエン酸 ０．１ｇ
クエン酸ソーダ １．０ｇ
パラオキシ安息香酸メチル ０．１ｇ
香料 ０．０５ｇ
精製水 残部（全量を1００ｍｌとする）
【０１０１】
実施例乳液と、スターフルーツの果実抽出物を含まない他は実施例乳液と同じ組成からなる比較例乳液とについて、下記の評価試験を行った。
２０〜４０代の女性２０名を５群、４名ずつに分け、朝夕１日２回、２週間、洗顔後に右顔面に実施例品を、左顔面に比較例品を塗布した。その際、使用時の感触、使用後のべたつき、保湿の持続性、肌荒れ改善効果の４項目について評価を行った。
【０１０２】
評価は、１〜５点の５段階評価によって行い、「５点」が「非常によい」（使用時の感触が非常によい、使用後のべたつきが全くない、保湿の持続性が非常に高い、肌荒れ改善効果が非常に高い）、「４点」が「よい」（使用時の感触がよい、使用後のべたつきがほとんどない、保湿の持続性が高い、肌荒れ改善効果が高い）、「３点」が「ふつう」（使用時の感触はふつう、使用後のべたつきはあまりない、保湿の持続性はある程度ある、肌荒れ改善効果はある程度ある）、「２点」が「ややわるい」（使用時の感触はややわるい、使用後のべたつきがややある、保湿の持続性はあまりない、肌荒れ改善効果はあまりない）、「１点」が「わるい」（使用時の感触がわるい、使用後のべたつきがかなりある、保湿の持続性が全くない、肌荒れ改善効果が全くない）とした。
評価点の平均を表９に示す。

【０１０３】
表９に示される官能評価によって、スターフルーツの果実抽出物が肌荒れ改善効果と優れた使用感とを有することが確認された。すなわち、スターフルーツの果実抽出物を配合した皮膚化粧料が皮膚の老化予防・改善作用（肌荒れ改善作用）を有するとともに、皮膚に適用した場合の使用感と安全性に優れていることが確認された。
【０１０４】
〔配合例１〕
下記の組成の乳液を常法により製造した。
スターフルーツ果実水抽出物 １ｇ
ホホバオイル ４ｇ
オリーブオイル ２ｇ
スクワラン ２ｇ
セタノール ２ｇ
モノステアリン酸グリセリル ２ｇ
ポリオキシエチレンセチルエーテル（20E.0） ２．５ｇ
オレイン酸ポリオキシエチレンソルビタン(20E.0) ２ｇ
黄杞エキス ０．１ｇ
イチョウ葉エキス ０．１ｇ
コンキオリン ０．１ｇ
オウバクエキス ０．１ｇ
カミツレエキス ０．１ｇ
１，３−ブチレングリコール ３ｇ
パラオキシ安息香酸メチル ０．１５ｇ
香料 ０．０５ｇ
精製水 残部（全量を１００ｇとする）
【０１０５】
〔配合例２〕
下記の組成の化粧水を常法により製造した。
スターフルーツ果実５０％エタノール抽出物 ２ｇ
グリセリン ３ｇ
１，３−ブチレングリコール ３ｇ
オレイン酸ポリオキシエチレンソルビタン(20E.0) ０．５ｇ
パラオキシ安息香酸メチル ０．１５ｇ
クエン酸 ０．１ｇ
クエン酸ソーダ ０．１ｇ
油溶性甘草エキス ０．１ｇ
海藻エキス ０．１ｇ
キシロビオースミクスチャー ０．５ｇ
クジンエキス ０．１ｇ
香料 ０．０５ｇ
精製水 残部（全量を１００ｇとする）
【０１０６】
〔配合例３〕
下記の組成のクリームを常法により製造した。
スターフルーツ果実エタノール抽出物 １ｇ
流動パラフィン ５ｇ
サラシミツロウ ４ｇ
セタノール ３ｇ
スクワラン １０ｇ
ラノリン ２ｇ
ステアリン酸 １ｇ
オレイン酸ポリオキシエチレンソルビタン(20E.0) １．５ｇ
モノステアリン酸グリセリル ３ｇ
１，３−ブチレングリコール ６ｇ
酵母抽出液 ０．１ｇ
シソ抽出液 ０．１ｇ
シナノキ抽出液 ０．１ｇ
ジユ抽出液 ０．１ｇ
パラオキシ安息香酸メチル １．５ｇ
香料 ０．１ｇ
精製水 残部（全量を１００ｇとする）
【０１０７】
〔配合例４〕
下記の組成のパックを常法により製造した。
スターフルーツ果実５０％エタノール抽出物 ５ｇ
ポリビニルアルコール １５ｇ
ポリエチレングリコール ３ｇ
プロピレングリコール ７ｇ
エタノール １０ｇ
セージ抽出液 ０．１ｇ
トウキ抽出液 ０．１ｇ
ニンジン抽出液 ０．１ｇ
パラオキシ安息香酸エチル ０．０５ｇ
香料 ０．０５ｇ
精製水 残部（全量を１００ｇとする）
【０１０８】
〔配合例５〕
下記の混合物を打錠して、錠剤状健康・栄養補助食品を製造した。
スターフルーツ果実水抽出物 ５０重量部
粉糖（ショ糖） １７８重量部
ソルビット １０重量部
グリセリン脂肪酸エステル １２重量部
【０１０９】
〔配合例６〕
下記の混合物を顆粒状に形成して栄養補助食品を製造した。
スターフルーツ果実エタノール抽出物 ３４重量部
ビートオリゴ糖 １０００重量部
ビタミンＣ １６７重量部
ステビア抽出物 １０重量部
【０１１０】
【発明の効果】
本発明により、保湿剤、抗酸化剤、抗老化剤、活性酸素消去剤、ラジカル消去剤、ヒアルロン酸産生促進剤、エラスターゼ阻害剤及びコラゲナーゼ阻害剤が提供される。また、本発明により、保湿作用、活性酸素消去作用、ラジカル消去作用、ヒアルロン酸産生促進作用、エラスターゼ阻害作用又はコラゲナーゼ阻害作用を有する皮膚化粧料及び美容用飲食品が提供される。
本発明の保湿剤、抗酸化剤、抗老化剤、活性酸素消去剤、ラジカル消去剤、ヒアルロン酸産生促進剤、エラスターゼ阻害剤及びコラゲナーゼ阻害剤、並びに皮膚化粧料及び美容用飲食品は、肌荒れ、皮膚の老化及びこれらに伴って生じる各種皮膚疾患を予防・改善する上で有用である。[0001]
BACKGROUND OF THE INVENTION
The present invention has a moisturizing action, active oxygen scavenging action, radical scavenging action, hyaluronic acid production promoting action, elastase inhibitory action or collagenase inhibitory action, and is a plant useful for preventing or improving rough skin or preventing or improving skin aging. The present invention relates to various drugs containing an extract, and skin cosmetics and beauty foods and drinks that are formulated with the plant extract to impart rough skin prevention / improvement action or skin aging prevention / improvement action.
[0002]
[Prior art]
The causes and onset mechanisms of rough skin and skin aging are diverse, but the main onset mechanisms and countermeasures are as follows.
[0003]
(1) Deterioration of moisture retention function
The skin's moisturizing function is reduced by aging and various environmental factors. Skin with reduced moisturizing function becomes dry and bulky and loses elasticity. Such dry skin may cause various skin diseases such as atopic dermatitis, and may cause aging of skin such as spots and wrinkles.
In order to prevent skin dryness, glycerin, propylene glycol, 1,3-butylene glycol and other polyhydric alcohols, as well as oil and fat ingredients, amino acids, proteins, polysaccharides, etc. have been used. Have problems in terms of feeling of use, durability of the moisturizing effect, safety, and the like.
[0004]
(2) Effects of active oxygen and in vivo radicals
In recent years, active oxygen has attracted attention as a factor that oxidizes biological components, and its adverse effect on living organisms has become a problem. Reactive oxygen is generated in the process of energy metabolism in living cells and is superoxide (ie, superoxide anion (· O₂-), Hydrogen peroxide (H₂O₂), Singlet oxygen (¹O₂) And hydroxy radicals (.OH). These active oxygens are essential for the phagocytic sterilization mechanism and play an important role in the removal of viruses and cancer cells. However, excessive generation of active oxygen attacks in vivo molecules that make up membranes and tissues in the body. Inducing various diseases. For example, active oxygen is thought to decompose, denature or crosslink biological tissues such as collagen, or to produce lipid peroxides that oxidize fats and oils and damage cells. It is thought that this disorder causes skin aging such as wrinkle formation and reduced elasticity.
[0005]
In the living body, radicals generated first based on oxygen are superoxide, and other radicals such as hydroxy radicals are generated via superoxide. Intracellular superoxide is converted to hydrogen peroxide by superoxide dismutase (SOD) produced in the cell, but the amount of SOD decreases with aging, and the decrease in SOD amount causes intracellular superoxide to enter the cell. The concentration increases and superoxide becomes damaging to the living body. For this reason, SOD itself, tocopherols, ougone extract, etc. are used as SOD-like agents to compensate for the decrease in the amount of SOD, but these SOD-like agents have problems in terms of safety and the like. .
[0006]
(3) Reduction / denaturation of extracellular matrix components
The dermis and epidermis of the skin are composed of epidermal cells, fibroblasts, and extracellular matrices such as elastin, collagen, and hyaluronic acid that are outside these cells and support the skin structure. By maintaining the constancy of the skin tissue interaction, moisture retention, flexibility, elasticity and the like are ensured, and the skin is maintained in a fresh state with a stretch and gloss. However, elastin, collagen, which are the main components of the extracellular matrix, due to the influence of certain external factors such as ultraviolet irradiation, drastic air drying, excessive skin washing, etc. The production amount of hyaluronic acid and the like is reduced, and denaturation and decomposition are caused. As a result, the stratum corneum begins to exfoliate abnormally, the skin loses its tension and gloss, and exhibits aging symptoms such as rough skin and wrinkles. In this way, changes accompanying aging of the skin, that is, wrinkles, dullness, disappearance of texture, decrease in elasticity, etc. involve reduction and degeneration of dermal extracellular matrix components such as elastin, collagen and hyaluronic acid. Yes.
[0007]
In recent years, involvement of matrix proteases has been pointed out as a factor inducing this change. Among the matrix proteases, collagenase, that is, MMP-1 (matrix metalloprotease-1), is known as an enzyme that degrades type I and III collagen, which are the main components of the dermal extracellular matrix of the skin. Is greatly increased by irradiation with ultraviolet rays, which contributes to decrease / denaturation of collagen due to ultraviolet rays and may be a major factor in the formation of wrinkles on the skin. Therefore, inhibition of collagenase activity is important in preventing and improving skin aging.
[0008]
[Problems to be solved by the invention]
Therefore, the present invention firstly finds a substance capable of preventing / ameliorating rough skin, aging of the skin, and various skin diseases associated therewith through a moisturizing action, and provides a moisturizing agent containing it as an active ingredient. For the purpose.
[0009]
Another object of the present invention is to find a substance capable of scavenging active oxygen and in vivo radicals through an active oxygen scavenging action or radical scavenging action, and to provide an antioxidant containing it as an active ingredient. And
[0010]
Furthermore, the present invention thirdly suppresses the decrease and degeneration of extracellular matrix constituents through hyaluronic acid production promoting action, elastase inhibitory action or collagenase inhibitory action, thereby causing rough skin, aging of the skin, and various types caused by these. An object of the present invention is to find a substance capable of preventing and ameliorating skin diseases and to provide an anti-aging agent containing it as an active ingredient.
[0011]
A fourth object of the present invention is to find a substance capable of scavenging active oxygen in the living body through a scavenging action of active oxygen, and to provide a reactive oxygen scavenger containing it as an active ingredient.
[0012]
A fifth object of the present invention is to provide a radical scavenger containing a substance capable of scavenging radicals in a living body through radical scavenging action and containing it as an active ingredient.
[0013]
Furthermore, the present invention sixthly provides a hyaluronic acid production promoter containing a substance capable of suppressing the decrease / denaturation of hyaluronic acid constituting the extracellular matrix through hyaluronic acid production promoting action and containing it as an active ingredient. The purpose is to provide.
[0014]
A seventh object of the present invention is to provide a elastase inhibitor containing a substance capable of suppressing the decrease / denaturation of elastin constituting an extracellular matrix through an elastase inhibitory action and containing the substance as an active ingredient. And
[0015]
Furthermore, an object of the present invention is to provide a collagenase inhibitor containing a substance capable of suppressing the decrease / denaturation of collagen constituting the extracellular matrix through a collagenase inhibitory action and containing it as an active ingredient. And
[0016]
In addition, the present invention ninthly has a moisturizing action, active oxygen scavenging action, radical scavenging action, hyaluronic acid production promoting action, elastase inhibitory action or collagenase inhibitory action, and occurs in association with rough skin, aging of the skin and these. The object is to provide a skin cosmetic useful for the prevention and improvement of various skin diseases.
[0018]
[Means for Solving the Problems]
In order to achieve the above object, the moisturizing agent, antioxidant, anti-aging agent, active oxygen scavenger, radical scavenger, hyaluronic acid production promoter, elastase inhibitor and collagenase inhibitor of the present invention include star fruit (Averrhoa carambola L.) extract from fruit as an active ingredient, and the skin cosmetics and cosmetics for food and drink of the present invention contain an extract from star fruit (Averrhoa carambola L.) fruit It is characterized by that. In the antioxidant of the present invention, the extract preferably has an active oxygen scavenging action and / or a radical scavenging action. In the anti-aging agent of the present invention, the extract preferably has one or more actions selected from the group consisting of hyaluronic acid production promoting action, elastase inhibiting action and collagenase inhibiting action.
[0019]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, the present invention will be described in detail.
In the present invention, the “extract from the fruit of a star fruit” includes an extract obtained from the fruit of a star fruit by an extraction process, a diluted or concentrated liquid of the extract, and a dry liquid obtained by drying the extract. Or any of these crudely purified products or purified products.
[0020]
What is used as an extraction raw material in the extraction process is a fruit of a star fruit (scientific name: Averrhoa carambola L.). Star fruit belongs to the Oxalis family, and is also called Goshiko or Yomo (herbal medicine name), and its fruit is edible. Star fruit has been described in Chinese literature since the BC, and its fruit is called star fruit because of its star-shaped cross section. Star fruit is cultivated in Okinawa, southeastern China, Yunnan and other tropical areas, and can be easily obtained from these areas.
[0021]
The fruit of the star fruit used as the extraction raw material is suitably dried and pulverized immediately after collection. Drying may be performed in the sun, or may be performed using a commonly used dryer. The fruit of the star fruit may be used as an extraction raw material after pretreatment such as degreasing with a nonpolar solvent such as hexane. By performing a pretreatment such as degreasing, the extraction treatment with the polar solvent of the fruit of the star fruit can be performed efficiently.
[0022]
In the extraction process, it is preferable to use a polar solvent as the extraction solvent. Moisturizing action, active oxygen scavenging action, radical scavenging action, hyaluronic acid production promoting action, elastase inhibitory action or collagenase inhibitory action ingredient contained in the fruit of star fruit can be easily extracted by extraction process using polar solvent as extraction solvent can do.
[0023]
Specific examples of suitable extraction solvents include water, lower aliphatic alcohols, hydrous lower aliphatic alcohols, and the like, and these can be used alone or as a mixture of two or more thereof. Specific examples of suitable lower aliphatic alcohols include methanol, ethanol, propanol, 1,3-butylene glycol, glycerin, propylene glycol, isoprene glycol and the like.
[0024]
Water that can be used as the extraction solvent includes pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, as well as those subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering, and the like. Therefore, the water that can be used as the extraction solvent in the present invention includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.
[0025]
When using the liquid mixture of 2 or more types of polar solvents as an extraction solvent, the mixing ratio can be adjusted suitably. For example, when using the liquid mixture of water and a lower aliphatic alcohol, the mixing ratio of water and a lower aliphatic alcohol can be 7: 3 to 2: 8 (weight ratio).
[0026]
The extraction treatment is not particularly limited as long as the soluble component contained in the fruit of the star fruit can be eluted in the extraction solvent, and can be performed according to a conventional method. In the extraction process, it is not necessary to adopt a special extraction method, and any apparatus can be used at room temperature or under reflux heating.
[0027]
For example, the extraction raw material is put into a treatment tank filled with the extraction solvent, and the soluble components are eluted with occasional stirring. At this time, although the extraction conditions can be appropriately adjusted according to the extraction raw material and the like, the amount of the extraction solvent is usually 5 to 15 times (weight ratio) of the extraction raw material, and the extraction time is usually 1 to 3 hours. The extraction temperature is usually from room temperature to 95 ° C.
[0028]
An eluate can be obtained by eluting soluble components by extraction and then filtering to remove the extraction residue. The obtained extract is diluted, concentrated, dried, purified, etc. according to a conventional method in order to obtain a diluted or concentrated solution of the extract, a dried product of the extract, or a crude purified product or a purified product thereof. Processing may be performed.
[0029]
The obtained extract can be used as it is as a moisturizer, antioxidant, anti-aging agent, active oxygen scavenger, radical scavenger, hyaluronic acid production promoter, elastase inhibitor and collagenase inhibitor. The liquid or its dried product is easier to use. In obtaining a dried extract, a carrier such as dextrin or cyclodextrin may be added to improve hygroscopicity. Moreover, the extract from the fruit of a star fruit can also be formulated according to a conventional method. When formulating, to facilitate storage and handling, add any pharmaceutically acceptable carrier such as dextrin, cyclodextrin or any other auxiliary agent, and add any agent such as powder, granules, tablets, etc. It can be formulated into a form.
[0030]
In addition, since the fruit of star fruit has a peculiar smell and taste, it can be purified for the purpose of decolorization, deodorization, etc. within the range that does not cause a decrease in its physiological activity. Since it is not used in large quantities when added to food and drink, there is no practical problem even if it is not purified. Specifically, purification can be performed by activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, or the like.
[0031]
The extract from the fruit of the star fruit obtained as described above has a moisturizing action, active oxygen scavenging action, radical scavenging action, hyaluronic acid production promoting action, elastase inhibitory action or collagenase inhibitory action. , Moisturizer, antioxidant, anti-aging agent, active oxygen scavenger, radical scavenger, hyaluronic acid production promoter, elastase inhibitor and collagenase inhibitor.
[0032]
The moisturizing agent of the present invention can prevent and / or improve rough skin, skin aging and various skin diseases caused by the skin moisturizing action.
[0033]
The antioxidant of the present invention eliminates active oxygen and in vivo radicals through an active oxygen scavenging action and / or radical scavenging action to prevent and / or improve rough skin, skin aging, and various skin diseases associated therewith. be able to. Here, in the present invention, “active oxygen” includes superoxide, hydrogen peroxide, hydroxy radical, singlet oxygen and the like. The term “radical” means a molecule or atom having one or more unpaired electrons, and includes superoxide, hydroxy radical, DPPH and the like.
[0034]
The anti-aging agent of the present invention suppresses reduction / denaturation of extracellular matrix components through one or more actions selected from the group consisting of hyaluronic acid production promoting action, elastase inhibitory action and collagenase inhibitory action, and rough skin It is possible to prevent and / or improve skin aging and various skin diseases caused thereby.
[0035]
The active oxygen scavenger of the present invention can eliminate active oxygen in a living body through an active oxygen scavenging action, and can prevent and / or improve rough skin, skin aging, and various skin diseases associated therewith.
[0036]
The radical scavenger of the present invention can scavenge radicals in the living body through radical scavenging action, and can prevent and / or improve rough skin, skin aging and various skin diseases associated therewith.
[0037]
The hyaluronic acid production promoter of the present invention activates the production of hyaluronic acid by fibroblasts to suppress hyaluronic acid decrease, degeneration, etc., and prevent rough skin, aging of the skin and various skin diseases caused by these And / or can be improved.
[0038]
The elastase inhibitor of the present invention can suppress and / or improve rough skin, aging of the skin, and various skin diseases caused by these by suppressing elastin reduction and degeneration by elastase through an elastase inhibitory action.
[0039]
The collagenase inhibitor of the present invention can suppress and / or improve rough skin, aging of the skin, and various skin diseases caused by these, by inhibiting collagen decrease and degeneration by collagenase through a collagenase inhibitory action.
[0040]
[Skin cosmetic]
The extract from the fruit of star fruit has moisturizing action, active oxygen scavenging action, radical scavenging action, hyaluronic acid production promoting action, elastase inhibitory action or collagenase inhibitory action. In addition to being able to prevent and / or ameliorate various skin diseases that occur as a result of this, it is suitable for blending into skin cosmetics because of its excellent usability and safety when applied to the skin. The skin cosmetic of the present invention includes any one or two of moisturizer, antioxidant, anti-aging agent, active oxygen scavenger, radical scavenger, hyaluronic acid production promoter, elastase inhibitor or collagenase inhibitor. You may mix | blend the above with skin cosmetics.
[0041]
The skin cosmetics that can be blended with the extract from the fruit of star fruit are not particularly limited, but specific examples thereof include ointments, creams, emulsions, lotions, packs, bathing agents, and the like.
[0042]
The compounding amount of the star fruit fruit extract in the skin cosmetic of the present invention can be adjusted as appropriate depending on the type of skin cosmetic, the physiological activity of the extract, etc., but it is about 0.00 when converted to a standard extract. It is preferable to mix | blend so that it may become 001-10 weight%.
[0043]
In the skin cosmetic of the present invention, the moisturizing action, active oxygen scavenging action, radical scavenging action, hyaluronic acid production promoting action, elastase inhibitory action or collagenase inhibitory action of the extract from the fruit of star fruit is not disturbed. Various main agents and auxiliaries usually used in the production of skin cosmetics and other optional auxiliaries can be used. The skin cosmetic composition of the present invention is not limited to the use of an extract from the fruit of star fruit alone as the main ingredient in relation to rough skin, aging of the skin, and various skin diseases associated therewith.
[0044]
Examples of skin cosmetics of the present invention that can be used as a skin cosmetic constituent together with an extract from the fruit of a star fruit include, for example, astringents, bactericides / antibacterial agents, whitening agents, ultraviolet absorbers, moisturizers, Examples include cell activators, anti-inflammatory / anti-allergic agents, antioxidant / active oxygen scavengers, etc. When the above components are used in combination, the synergistic effects with the components used in combination are superior to those normally expected. May have a positive effect.
[0045]
When producing skin cosmetics containing an extract from the fruit of star fruit, the choice of other production raw materials is rarely limited, and oils, waxes, hydrocarbons, fatty acids, alcohols, Any of general substrates and auxiliaries such as esters, surfactants, and fragrances can be used.
[0046]
[Beauty food and drink]
The extract from the fruit of star fruit has moisturizing action, active oxygen scavenging action, radical scavenging action, hyaluronic acid production promoting action, elastase inhibitory action or collagenase inhibitory action. In addition to being able to prevent and / or ameliorate various skin diseases that occur as a result, and can be taken orally, it is suitable for blending into any food or beverage supplement or dietary supplement. In this case, it is appropriate that the amount of the extract per day for adults is about 1 to 1000 mg in consideration of the general intake of the food and drink to be added.
[0047]
Foods and drinks containing extracts from the fruits of star fruit are given prevention and improvement of rough skin, aging of the skin and various skin diseases that accompany these, and can be used as cosmetic foods and drinks. . Here, “beauty food and drink” refers to food and drink intended for beautifying the skin, or food and drink intended to prevent and improve rough skin, aging of the skin and various skin diseases associated therewith. Means.
[0048]
The cosmetic food and drink of the present invention may be a blend of an extract from the fruit of a star fruit in any food or drink that does not impede its activity, or a nutritional supplement comprising the extract as a main component. It may be food.
[0049]
When producing the cosmetic food and drink of the present invention, for example, sugars such as dextrin and starch; proteins such as gelatin, soybean protein and corn protein; amino acids such as alanine, glutamine and isoleucine; cellulose and gum arabic Polysaccharides: It can be formulated into an arbitrary dosage form by adding optional auxiliaries such as soybean oil and oils such as medium chain fatty acid triglycerides.
[0050]
Foods and drinks that can be blended with the extract from the fruit of the star fruit are not particularly limited. Specific examples thereof include beverages such as soft drinks, carbonated drinks, nutritional drinks, fruit drinks, and lactic acid drinks (concentrated liquids of these drinks and Ice cream, ice sherbet, shaved ice, etc .; noodles such as buckwheat, udon, harusame, gyoza skin, shumai skin, Chinese noodles, instant noodles; candy, candy, gum, chocolate, snacks , Biscuits, jelly, jam, cream, baked confectionery, etc .; fish and livestock processed foods such as kamaboko, ham, sausage; dairy products such as processed milk, fermented milk; salad oil, tempura oil, margarine, mayonnaise, shortening, Fats and oils such as whipped cream and dressings; processed foods such as sauces; sauces, sauces, etc .; tablets, granules, etc. Health and nutritional supplements such people of form; and other soups, stews, salads, prepared foods, pickles, and the like.
[0051]
The moisturizer, antioxidant, anti-aging agent, active oxygen scavenger, radical scavenger, hyaluronic acid production promoter, elastase inhibitor, collagenase inhibitor, skin cosmetic and cosmetic food and drink of the present invention described above are Although it is suitably applied to humans, it can also be applied to animals other than humans as long as each effect is exhibited.
[0052]
【Example】
Hereinafter, although a manufacture example, a test example, and a compounding example are shown and this invention is demonstrated concretely, this invention is not limited to each following example at all.
[0053]
[Production Example 1]
100 g of a coarsely pulverized fruit of a star fruit (scientific name: Averrhoa carambola L.) was put into 1000 ml of an extraction solvent, and extracted with gentle stirring at 80 ° C. for 3 hours. Thereafter, the mixture was filtered, and the filtrate was concentrated under reduced pressure at 40 ° C., and further dried with a vacuum dryer to obtain a fruit extract of star fruit. When the above extraction treatment was performed using three types of extraction solvents, the yield of the extract was as shown in Table 1. When the extraction solvent is a mixture, the mixing ratio shown below is based on weight.
[0054]
[Table 1]
Sample      Extraction solvent      Extract yield (wt%)
1 water 17.7
2 Ethanol / water (1/1) 20.5
3 Ethanol 12.8
[0055]
[Test Example 1] Moisturizing effect test
The samples 1 to 3 obtained in Production Example 1 were tested for their moisturizing effect by the following method.
A 0.01% aqueous solution of sample 1 to 3 (sample solution 1 to 3), 1% glycerin (sample solution 4) and purified water (sample solution 5) were each a paper disk (manufactured by Toyo Manufacturing Co., Ltd., weight: 10 μl was added dropwise to about 0.017 g). This was left in the test chamber, and the weight after 0 to 8 minutes was measured every minute. The water residual ratio (%) of each sample solution was determined by setting the weight of 0 minute to 100%. The test was performed at room temperature of 20 ° C. and humidity of 65%.
Table 2 shows the water residual ratio (%) of each sample solution after 0 to 8 minutes.
[0056]

[0057]
As shown in Table 2, the sample solution containing the fruit extract of the star fruit (sample solutions 1 to 3) is purified water (sample) in the same manner as the sample solution containing glycerin (humectant) (sample solution 4). The water residual rate was higher than that of the solution 5). From this, it was confirmed that the fruit extract of star fruit has a moisturizing action.
[0058]
[Test Example 2] Superoxide scavenging action test (NBT method)
Samples 1 to 3 obtained in Production Example 1 were tested for their superoxide scavenging action by the following method.
3 mM xanthine, 3 mM EDTA, 1.5 mg / mL BSA solution and 0.75 mM nitroblue tetrazolium (NBT) 0.1 mL each, 0.05 M Na₂CO_Three2.4 mL of buffer solution (pH 10.2) was placed in a test tube, and 0.1 mL of the sample solution was added thereto and left at 25 ° C. for 10 minutes. Next, 0.1 mL of a xanthine oxidase solution was added, stirred rapidly, and allowed to stand at 25 ° C. for 20 minutes. Thereafter, 0.1 mL of 6 mM copper chloride was added to stop the reaction, and the absorbance at a wavelength of 560 nm was measured. Hereinafter, this absorbance is referred to as “absorbance upon addition of sample solution and enzyme solution”.
[0059]
The same operation and measurement of absorbance were performed without adding the enzyme solution. The absorbance measured at this time is referred to as “absorbance when the sample solution is added and the enzyme solution is not added”.
[0060]
The same measurement was performed when distilled water was added without adding the sample solution. The absorbance measured at this time is referred to as “absorbance when no sample solution is added and the enzyme solution is added”.
[0061]
The same measurement was performed when distilled water was added without adding the enzyme solution and without adding the sample solution. The absorbance measured at this time is referred to as “absorbance when no sample solution is added and no enzyme solution is added”.
And the superoxide elimination rate was calculated | required by following Formula.
[0062]
Erase rate (%) = {1− (A−B) / (C−D)} × 100
[0063]
In the above formula, “A” is “absorbance when sample solution is added and enzyme solution is added”, “B” is “absorbance when sample solution is added and enzyme solution is not added”, and “C” is “sample solution is not added and enzyme is added” “Absorptivity when solution is added” and “D” represent “absorbance when sample solution is not added and enzyme solution is not added”.
[0064]
The above-mentioned erasure rate was measured by decreasing the sample concentration stepwise, and the sample concentration (ppm; μg / mL) at which the superoxide erasure rate was 50% was determined by interpolation. The results are shown in Table 3.
[0065]
[Table 3]
sample NO.   Extract    50% erased sample concentration (ppm)
1 Water extract 19.0
2 50% ethanol extract 16.9
3 Ethanol extract 25.3
[0066]
From the results shown in Table 3, it was confirmed that the fruit extract of star fruit has a superoxide scavenging action. It was also confirmed that the degree of superoxide scavenging action can be adjusted by the concentration of the extract.
[0067]
[Test Example 3] Radical scavenging action test for DPPH
Samples 1 to 3 obtained in Production Example 1 were tested for their radical scavenging action using DPPH, which is a very stable radical, by the following method.
1.5 × 10^-Four3 mL of the sample solution was added to 3 mL of the MDPPH ethanol solution, and the container was immediately sealed and shaken and allowed to stand for 30 minutes. Thereafter, absorbance at a wavelength of 520 nm was measured. As a control, the same operation was performed using a solvent in which the sample solution was dissolved instead of the sample solution, and the absorbance at a wavelength of 520 nm was measured. As a blank, the absorbance at a wavelength of 520 nm was measured immediately after 3 mL of the sample solution was added to ethanol. From each measured absorbance, the radical scavenging rate (%) was calculated by the following formula.
[0068]
Erase rate (%) = {1− (B−C) / A} × 100
[0069]
In the above formula, “A” represents “control absorbance”, “B” represents “absorbance when a sample solution is added”, and “C” represents “blank absorbance”.
[0070]
The above-mentioned erasure rate was measured by gradually decreasing the sample concentration, and the sample concentration (ppm; μg / mL) at which the DPPH radical erasure rate was 50% was determined by interpolation. The results are shown in Table 4.
[0071]
[Table 4]
sample NO.   Extract    50% erased sample concentration (ppm)
1 Water extract 25.0
2 50% ethanol extract 26.0
3 Ethanol extract 28.5
[0072]
From the results shown in Table 4, it was confirmed that the fruit extract of star fruit has a radical scavenging action. It was also confirmed that the degree of radical scavenging action can be adjusted by the concentration of the extract.
[0073]
[Test Example 4] Singlet oxygen scavenging action test
Samples 1 to 3 obtained in Production Example 1 were tested for singlet oxygen scavenging action by the following method.
5 mL of 2% red blood cell suspension in a clear glass bottle (10 mL volume), 5 mL of isotonic phosphate buffer pH 7.4 containing a sample at a predetermined concentration, and photosensitizer (10 mM hematoporphyrin-20 mM sodium hydroxide solution) ) 0.01 mL was mixed. The resulting solution was uniformly irradiated on a merry-go-round with a 7.5 W halogen lamp for 35 minutes to produce singlet oxygen (¹O₂) To cause hemolysis of red blood cells. 1 mL of the reaction solution was collected, 2 mL of isotonic phosphate buffer was added and mixed, and then centrifuged at 4 ° C. and 3000 rpm for 5 minutes. The supernatant was then collected and the absorbance at a wavelength of 540 nm was measured. Separately, 1 mL of the reaction solution in which red blood cells were partially hemolyzed was taken, and 2 mL of distilled water was added thereto to completely hemolyze it, and the absorbance was measured in the same manner. The singlet oxygen scavenging rate was determined from the measured absorbance by the following formula.
[0074]
Singlet oxygen elimination rate (%) = (1−B / A) × 100
[0075]
In the above formula, “A” represents “absorbance of control” and “B” represents “absorbance of reaction solution supernatant”.
[0076]
The above-mentioned erasure rate was measured by decreasing the sample concentration stepwise, and the sample concentration (ppm; μg / mL) at which the singlet oxygen erasure rate was 50% was determined by interpolation. The results are shown in Table 5.
[0077]
[Table 5]
sample NO.   Extract    50% erased sample concentration (ppm)
1 Water extract 147.3
2 50% ethanol extract 122.5
3 Ethanol extract 118.4
[0078]
From the results shown in Table 5, it was confirmed that the fruit extract of star fruit has a singlet oxygen scavenging action. It was also confirmed that the degree of singlet oxygen scavenging action can be adjusted by the concentration of the extract.
[0079]
[Test Example 5] Hyaluronic acid production promoting action test
Samples 1 to 3 obtained in Production Example 1 were tested for hyaluronic acid production promoting effect by the following method.
Normal human neonatal fibroblasts (NB1RGB) 1 × 10⁶75cm²Using a flask in an α-MEM medium (GIBCO) (pH 7.2) containing 10% FBS at 37 ° C., 5% CO₂Cultured for 7 days under -95% air. Cells were collected by trypsinization and 2.2 × 10 6 using α-MEM medium containing 1% FBS.^FourPer microliter and seeded 100 μL each in a 96-well microplate at 37 ° C., 5% CO₂Cultured overnight under -95% air. On the next day, 100 μL of α-MEM medium containing 1% FBS in which a sample (sample concentration: 50 ppm or 200 ppm) (ppm = μg / mL) was dissolved was added to each well at 37 ° C., 5% CO₂-Cultured for 3 days under -95% air.
[0080]
10 μL of the culture supernatant was diluted 10-fold with 90 μL of PBS (−), 50 μL thereof was added to an ELISA plate previously coated with hyaluronic acid, and ELISA was performed using various antibodies. Hyaluronic acid was quantified using a calibration curve. The hyaluronic acid production promotion rate was determined with the value when no sample was added as 100%. The results are shown in Table 6.
[0081]

[0082]
From the results shown in Table 6, it was confirmed that the fruit extract of star fruit has hyaluronic acid production promoting action. It was also confirmed that the degree of hyaluronic acid production promoting action can be adjusted by the concentration of the extract.
[0083]
[Test Example 6] Elastase inhibitory action test
Samples 1 to 3 obtained in Production Example 1 were tested for their elastase inhibitory action by the following method.
A 96-well plate was prepared, and 50 μL of a sample solution (solvent: DMSO + water) and 50 μL of an elastase solution were added to one well, and 100 μL of a substrate solution was further added and mixed. After reacting at 25 ° C. for 15 minutes, absorbance at a wavelength of 415 nm was measured. Enzymatic reaction and absorbance measurement similar to the above were performed by adding only the solvent equivalent to the sample solution instead of the sample solution. Further, in each case, the same operation and measurement were performed by adding a buffer instead of the elastase solution.
[0084]
The elastase solution used was a solution obtained by dissolving 5 mg of Sigma Elastase Type III in 1 mL of 0.2 mol / L Tris-HCl buffer at pH 8 and diluting it 250 times. As a substrate solution, a solution having a concentration of 45.14 mg / mL in which N-SUCCINYL-ALA-ALA-ALA p-NITROANILIDE (Sigma) was dissolved in DMSO was diluted 100-fold with the above Tris-HCl buffer.
From the measurement results, the elastase inhibition rate (%) was determined based on the following formula.
[0085]
Elastase inhibition rate (%) = [1- (A−B) / (C−D)] × 100
[0086]
In the above formula, “A” is the absorbance when the sample solution is added and the enzyme is added, “B” is the absorbance when the sample solution is added and no enzyme is added, “C” is the absorbance when no sample is added and the enzyme is added, and “D” Represents the absorbance when no sample was added and no enzyme was added.
[0087]
The inhibition rate was measured by gradually reducing the sample concentration, and the concentration of the sample solution (ppm; μg / mL) that inhibits the elastase activity by 50% was determined by interpolation.
The test results are shown in Table 7.
[0088]
[Table 7]
sample NO.   Extract    50% inhibitory sample concentration (ppm)
1 Water extract 200
2 50% ethanol extract 182
3 Ethanol extract 175
[0089]
As shown in Table 7, it was confirmed that the fruit extract of star fruit has an elastase inhibitory action. In addition, it was confirmed that the elastase inhibitory action level can be adjusted by the concentration of the extract.
[0090]
[Test Example 7] Test for collagenase inhibitory action
The samples 1 to 3 obtained in Production Example 1 were tested for their collagenase inhibitory action by the following method.
50 μL of a sample solution (solvent: 20 mol / L calcium chloride-containing 0.1 mol / L Tris-HCl buffer (pH 7.1): the same in the following buffer), 50 μL of collagenase solution and 400 μL of substrate solution were mixed at 37 ° C. Incubated for 30 minutes. The reaction was then stopped with 1 mL of a 25 mmol / L citric acid solution and extracted with 5 mL of ethyl acetate. About the obtained extract, the light absorbency (control liquid: ethyl acetate) of wavelength 320nm was measured. The absorbance measured at this time is referred to as “absorbance upon addition of enzyme or sample solution”. The collagenase solution used was a solution obtained by dissolving 5 mg of Sigma's collagenase Type IV in 1 mL of buffer and diluting it 50 times at the time of use. For the substrate solution, the BACHEM Fenchemikaline AG Pz-peptide was dissolved in the above buffer so as to have a concentration of 0.5 mol / L.
[0091]
In addition, the same enzyme reaction and absorbance measurement as described above were performed by adding an equal amount of buffer solution to the sample solution instead of the sample solution. The absorbance measured at this time is referred to as “absorbance when enzyme is added but no sample solution is added”.
[0092]
In addition, the same enzyme reaction and absorbance measurement as described above were performed by adding a buffer solution instead of the collagenase solution. The absorbance measured at this time is referred to as “absorbance when no enzyme is added and the sample solution is added”.
[0093]
In addition, the same enzyme reaction and absorbance measurement as described above were performed by adding a buffer solution equivalent to the sample solution instead of the sample solution and adding a buffer solution instead of the collagenase solution. The absorbance measured at this time is referred to as “absorbance when no enzyme is added and no sample solution is added”.
And collagenase inhibition rate (%) was computed by following Formula.
[0094]
Collagenase inhibition rate (%) = {1− (A−B) / (C−D)} × 100
[0095]
In the above formula, “A” is “absorbance when enzyme is added and sample solution is added”, “B” is “absorbance when enzyme is not added and sample solution is added”, and “C” is “when enzyme is added and sample solution is not added” "D" and "D" represent "absorbance when no enzyme is added and sample solution is not added".
[0096]
The inhibition rate was measured by gradually reducing the sample concentration, and the sample concentration (ppm; μg / mL) that inhibits the collagenase activity by 50% was determined by interpolation. The results are shown in Table 8.
[0097]
[Table 8]
sample NO.   Extract    50% inhibitory sample concentration (ppm)
1 Water extract 68.0
2 50% ethanol extract 53.2
3 Ethanol extract 50.1
[0098]
From the results shown in Table 8, it was confirmed that the fruit extract of star fruit has a collagenase inhibitory action. It was also confirmed that the degree of collagenase inhibitory action can be adjusted by the concentration of the extract.
[0099]
[Test Example 8] Testing of rough skin improvement (skin aging prevention / improvement)
A milky lotion (hereinafter referred to as “Example milky lotion”) containing a 50% ethanol extract (sample 2) from the fruit of the star fruit obtained in Production Example 1 was prepared according to a conventional method. The composition of the example emulsion is shown below.
[0100]
0.1 g of star fruit fruit extract (Sample 2 of Production Example 1)
Cetyl alcohol 0.5g
Beeswax 2.0g
1,3-butylene glycol 3.0g
Oleic acid polyoxyethylene sorbitan (20E.0) 0.5g
Hyaluronic acid 0.1g
Citric acid 0.1g
Sodium citrate 1.0g
Methyl paraoxybenzoate 0.1g
Fragrance 0.05g
Purified water remainder (total volume is 100 ml)
[0101]
The following evaluation test was performed on the Example emulsion and the Comparative Example emulsion having the same composition as the Example emulsion except that it did not contain the fruit extract of the star fruit.
Twenty women in their 20s and 40s were divided into 5 groups and 4 people, and the example product was applied to the right face and the comparative example product was applied to the left face twice a day in the morning and evening for 2 weeks. At that time, four items were evaluated: feel during use, stickiness after use, durability of moisture retention, and effect of improving skin roughness.
[0102]
The evaluation is performed by a five-step evaluation of 1 to 5 points. “5 points” is “very good” (the feel at the time of use is very good, there is no stickiness after use, and the durability of moisturizing is very high. ”4 points” is “good” (good feel during use, little stickiness after use, high moisturizing persistence, high skin roughness improvement effect), “3” "Dot" is "normal" (feels normal when used, there is not much stickiness after use, there is a certain degree of moisturizing persistence, and there is a degree of rough skin improvement effect), "2 points" is "slightly bad" (during use) The touch of the product is slightly bad, the stickiness after use is slightly, the persistence of moisturizing is not so much, the effect of improving the rough skin is not so much, and the "1 point" is "bad" (the touch is bad when using, the sticky after use) There is considerable, there is no persistence of moisturizing, rough skin Improvement effect was absolutely no).
Table 9 shows the average of the evaluation points.

[0103]
The sensory evaluation shown in Table 9 confirmed that the fruit extract of star fruit had a rough skin improvement effect and excellent usability. That is, it was confirmed that skin cosmetics containing a fruit extract of star fruit have an anti-aging / improving effect on skin aging (an effect of improving rough skin), and are excellent in usability and safety when applied to the skin. It was.
[0104]
[Formulation Example 1]
An emulsion having the following composition was produced by a conventional method.
1g fruit extract from star fruit
Jojoba oil 4g
2g olive oil
Squalane 2g
Cetanol 2g
2g glyceryl monostearate
Polyoxyethylene cetyl ether (20E.0) 2.5g
Oleic acid polyoxyethylene sorbitan (20E.0) 2g
Twilight extract 0.1g
Ginkgo biloba extract 0.1g
Conchiolin 0.1g
Oat extract 0.1g
Chamomile extract 0.1g
1,3-butylene glycol 3g
Methyl paraoxybenzoate 0.15g
Fragrance 0.05g
Purified water remainder (total amount is 100 g)
[0105]
[Formulation Example 2]
A lotion having the following composition was produced by a conventional method.
2g 50% ethanol extract of star fruit
Glycerin 3g
1,3-butylene glycol 3g
Oleic acid polyoxyethylene sorbitan (20E.0) 0.5g
Methyl paraoxybenzoate 0.15g
Citric acid 0.1g
Sodium citrate 0.1g
Oil soluble licorice extract 0.1g
Seaweed extract 0.1g
Xylobiose Mixture 0.5g
Kujin extract 0.1g
Fragrance 0.05g
Purified water remainder (total amount is 100 g)
[0106]
[Composition Example 3]
A cream having the following composition was produced by a conventional method.
Starfruit fruit ethanol extract 1g
Liquid paraffin 5g
Salami beeswax 4g
Setanol 3g
Squalane 10g
Lanolin 2g
Stearic acid 1g
1.5 g of polyoxyethylene sorbitan oleate (20E.0)
3g glyceryl monostearate
1,3-butylene glycol 6g
Yeast extract 0.1g
Perilla extract 0.1g
Linden extract 0.1g
Jiuyu Extract 0.1g
1.5 g of methyl paraoxybenzoate
Fragrance 0.1g
Purified water remainder (total amount is 100 g)
[0107]
[Formulation Example 4]
A pack having the following composition was produced by a conventional method.
5g ethanol extract of 50% star fruit
Polyvinyl alcohol 15g
Polyethylene glycol 3g
7g of propylene glycol
Ethanol 10g
Sage extract 0.1g
Toki extract 0.1g
Carrot extract 0.1g
0.05 g ethyl paraoxybenzoate
Fragrance 0.05g
Purified water remainder (total amount is 100 g)
[0108]
[Formulation Example 5]
The following mixture was tableted to produce a tablet-like health / nutritional supplement.
50 parts by weight of star fruit fruit water extract
178 parts by weight of powdered sugar (sucrose)
10 parts by weight of sorbit
Glycerin fatty acid ester 12 parts by weight
[0109]
[Composition Example 6]
The following mixture was formed into granules to produce a dietary supplement.
34 parts by weight of starfruit fruit ethanol extract
1000 parts by weight of beet oligosaccharide
167 parts by weight of vitamin C
Stevia extract 10 parts by weight
[0110]
【The invention's effect】
According to the present invention, there are provided a humectant, an antioxidant, an anti-aging agent, an active oxygen scavenger, a radical scavenger, a hyaluronic acid production promoter, an elastase inhibitor and a collagenase inhibitor. In addition, according to the present invention, skin cosmetics and cosmetic foods and drinks having a moisturizing action, active oxygen scavenging action, radical scavenging action, hyaluronic acid production promoting action, elastase inhibiting action or collagenase inhibiting action are provided.
The moisturizer, antioxidant, anti-aging agent, active oxygen scavenger, radical scavenger, hyaluronic acid production promoter, elastase inhibitor and collagenase inhibitor of the present invention, skin cosmetics and cosmetic foods and drinks are rough skin, It is useful for preventing and improving skin aging and various skin diseases caused by these.

Claims (11)

  A moisturizer characterized by containing an extract from the fruit of star fruit (Averrhoa carambola L.) as an active ingredient.   An antioxidant comprising an extract from the fruit of star fruit (Averrhoa carambola L.) as an active ingredient.   The antioxidant according to claim 2, wherein the extract has an active oxygen scavenging action and / or a radical scavenging action.   An anti-aging agent comprising an extract from the fruit of star fruit (Averrhoa carambola L.) as an active ingredient.   5. The anti-aging agent according to claim 4, wherein the extract has one or more actions selected from the group consisting of hyaluronic acid production promoting action, elastase inhibitory action and collagenase inhibitory action.   An active oxygen scavenger characterized by containing, as an active ingredient, an extract from the fruit of star fruit (Averrhoa carambola L.).   A radical scavenger characterized by containing an extract from the fruit of star fruit (Averrhoa carambola L.) as an active ingredient.   Hyaluronic acid production promoter characterized by containing an extract from the fruit of star fruit (Averrhoa carambola L.) as an active ingredient.   An elastase inhibitor characterized by containing an extract from the fruit of star fruit (Averrhoa carambola L.) as an active ingredient.   A collagenase inhibitor characterized by containing an extract from the fruit of star fruit (Averrhoa carambola L.) as an active ingredient.   Skin cosmetic characterized by containing an extract from the fruit of star fruit (Averrhoa carambola L.).