JP4542765B2 - 酵素阻害剤 - Google Patents
酵素阻害剤 Download PDFInfo
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- JP4542765B2 JP4542765B2 JP2003371316A JP2003371316A JP4542765B2 JP 4542765 B2 JP4542765 B2 JP 4542765B2 JP 2003371316 A JP2003371316 A JP 2003371316A JP 2003371316 A JP2003371316 A JP 2003371316A JP 4542765 B2 JP4542765 B2 JP 4542765B2
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Description
また、本発明は、オオウメガサソウを含むイチヤクソウ科の植物、例えばイチヤクソウ科ウメガサソウ属、特にオオウメガサソウを含有するα-アミラーゼ阻害作用、α-グルコシダーゼ阻害作用等を含むα-1,4-グルコシド結合分解酵素阻害作用、リパーゼ阻害作用が強い酵素阻害剤に関し、該酵素阻害剤を含有する食品組成物、化粧料組成物、医薬部外品組成物又は医薬組成物に関する。
さらに、本発明はオオウメガサソウを含むイチヤクソウ科の植物、例えばイチヤクソウ科ウメガサソウ属、特にオオウメガサソウを含有するα-アミラーゼ阻害作用並びにα-グルコシダーゼ阻害作用等を含むα-1,4-グルコシド結合分解酵素阻害作用、リパーゼ阻害作用が強い酵素阻害剤の製造方法に関する。
本発明は、オオウメガサソウを含むイチヤクソウ科の植物、例えばイチヤクソウ科ウメガサソウ属、特にオオウメガサソウを含有する酵素阻害効果を用いた方法に関する。つまり、該酵素阻害効果を用いた食品組成物、化粧料組成物、医薬部外品組成物、医薬組成物等の組成物に関する。
しかし、現在までに安全でかつ副作用の少ない植物抽出物を含有するα-アミラーゼ阻害剤、α-グルコシダーゼ阻害剤、リパーゼ阻害剤について知られているが、その効果について十分でなく、さらに高い効果を奏する予防、治療剤が望まれている。
本発明は、オオウメガサソウを含むイチヤクソウ科の植物、例えばイチヤクソウ科ウメガサソウ属、特にオオウメガサソウを含有する食品組成物、化粧料組成物、医薬部外品組成物又は医薬組成物を提供する。
さらに、本発明はオオウメガサソウを含むイチヤクソウ科の植物、例えばイチヤクソウ科ウメガサソウ属、特にオオウメガサソウを含有するα-1,4-グルコシド結合分解酵素阻害効果及び/又はリパーゼ阻害効果を有する酵素阻害剤の製造方法に関する。
本発明は、オオウメガサソウを含むイチヤクソウ科の植物、例えばイチヤクソウ科ウメガサソウ属、特にオオウメガサソウを含有するα-1,4-グルコシド結合分解酵素阻害効果及び/又はリパーゼ阻害効果を有する酵素阻害効果を用いた方法に関する。つまり、該酵素阻害効果を用いた食品組成物、化粧料組成物、医薬部外品組成物、医薬組成物等の組成物に関する。
本発明のオオウメガサソウは、イチヤクソウ科ウメガサソウ属オオウメガサソウ(PIPSISSEWA;Chimaphila umbellata (L.) W.P.C.Barton)であり、イチヤクソウ科の植物で海岸に近い乾燥した林中に自生する多年生の草本である。北アメリカ、ヨーロッパ、シベリア等の温帯に産し、日本では茨城県以北に分布している。本発明のオオウメガサソウは、アメリカにおいては古くからハーブティーやキャンディの香料として使用され、ヒトに対して長年使用されており安全性の高い植物である。
本発明のオオウメガサソウを含むイチヤクソウ科の植物は、該イチヤクソウ科の植物の全草又は葉、茎、根、種子等植物体の任意の各部位を用いることができる。該イチヤクソウ科の植物をそのまま用いてもよいし、これらを乾燥した後粉砕して粉末状にして用いることもできる。
本発明のオオウメガサソウを含むイチヤクソウ科の植物、特にオオウメガサソウを含有するα-アミラーゼ阻害作用並びに/若しくはα-グルコシダーゼ阻害作用等を含むα-1,4-グルコシド結合分解酵素阻害作用及び/又はリパーゼ阻害作用を有する酵素阻害剤を含む化粧料組成物、医薬部外品組成物、医薬組成物は、上記本発明のオオウメガサソウを含むイチヤクソウ科の植物、特にオオウメガサソウを少なくとも含み、該酵素阻害剤の配合量は、その剤型等について必要に応じ、考慮して適宜定めることが好ましい。好ましくは、上記オオウメガサソウを含むイチヤクソウ科の植物、特にオオウメガサソウを、全量に対して0.001〜50重量%、より好ましくは、0.01〜20重量%程度とすればよい。
本発明の確立により、糖尿病予防効果、糖尿病治療効果、肥満予防効果、肥満治療効果、高脂血症予防効果、高脂血症治療効果、動脈硬化予防効果、動脈硬化治療効果、フケ予防効果、アクネ予防効果が著しく促進され、安全性が高く、使用性に優れた酵素阻害剤等を提供可能となり、これが与える影響は広く、とりわけ、食品、化粧料、医薬部外品、医薬品等の各種産業界に与える工業的意義はきわめて高い。
本発明のオオウメガサソウを含むイチヤクソウ科の植物、特にオオウメガサソウを含有する組成物は、基本的には、食品材料から構成され、毒性を懸念することなく利用できる。
以下、実施例によって本発明を具体的に説明するが、本発明は、以下の実施例によって限定されるものではない。
酵素としてブタ膵アミラーゼを用い、ヨウ素デンプン反応に基づくアミラーゼテストワコー(和光純薬社製)でオオウメガサソウのα-アミラーゼ阻害活性を測定した。すなわち、基質である40mg/dLでんぷん250μlと、オオウメガサソウ抽出物(実施例2)を溶解した250mMリン酸ナトリウム緩衝液(pH7.0)100μlを混和し、37℃、6分間プレインキュベートした。その後、ブタ膵臓由来α-アミラーゼ溶液(2.5unit/ml)を10μl加え、37℃、7.5分間インキュベートした。更に、発色試薬(0.01Nヨウ素液)250μlと純水1,390μlを加え、1時間以内に660nmで吸光度を測定した。別に、対照として試料溶液の代わりに緩衝液を、またブランクとして試料溶液と酵素溶液の代わりに緩衝液を、それぞれ加え同様に操作して吸光度を測定した。また、α-アミラーゼ、リパーゼ阻害作用を持つ事が知られているイチヤクソウ科イチヤクソウ属の植物であるイチヤクソウ抽出物(以下の作成方法により製造)、ビワ葉抽出物(市販品)、羅布麻抽出物(市販品)、荷葉抽出物(市販品)を試験に用いた。
なお、イチヤクソウ科イチヤクソウ属イチヤクソウ(Pyrola japonica klenze ex Alef)に関し、全草100gに、50%エタノール1,000mlを加え、室温で1時間浸漬抽出した後、吸引濾過し、該イチヤクソウの抽出液を得た。これを減圧濃縮乾固して、60℃で12時間以上加熱乾燥したことによりイチヤクソウ抽出物の粉末を得た。
酵素としてブタ膵リパーゼ(ナカライテスク社製)を用い、S-アシル化合物を基質としたリパーゼキットS(大日本製薬社製)でオオウメガサソウのリパーゼ阻害活性を測定した。すなわち、オオウメガサソウの抽出物(実施例2)を溶解した試料溶液150μl、酵素溶液(ブタ膵リパーゼ0.05mg/ml;125mMトリス塩酸(pH7.4)に溶解)40μlと発色液(5,5´-ジチオビス(2−ニトロ安息香酸)0.1mg/ml)500μlを混和し、次にエステラーゼ阻害液(フェニルメチルスルホニルフルオリド3.48mg/ml)10μlを加え、30℃、5分間プレインキュベートした。その後、基質液(三酪酸ジメルカプロール6.69mg/ml及びドデシル硫酸ナトリウム5.73mg/ml)50μlを加え、暗所で30℃、30分間反応した。その後、反応停止液1mlを添加し、412nmで吸光度を測定した。別に、対照として試料溶液の代わりに緩衝液を、またブランクとして試料溶液と酵素溶液の代わりに緩衝液を、それぞれ加え以下の同様に操作して吸光度を測定した。対照として、α-アミラーゼ、リパーゼ阻害作用を持つ事が知られているイチヤクソウ科イチヤクソウ属の植物であるイチヤクソウ抽出物(実施例3で得たイチヤクソウ抽出物)、ビワ葉抽出物(市販品)、羅布麻抽出物(市販品)、荷葉抽出物(市販品)を試験に用いた。
α-グルコシダーゼ(シグマ社製)とp-ニトロフェニル-α-D-グルコピラノシド(ナカライテスク社製)の反応で生成するp-ニトロフェノールの量を比色定量で測定し、オオウメガサソウのα-グルコシダーゼ阻害活性を測定した。すなわち、オオウメガサソウの抽出物(実施例2)を溶解した試料溶液10μl、α-グルコシダーゼ溶液(from Brewers Yeast 800mU/ml;50mMリン酸カリウム緩衝液(pH7.0)、100mM NaClに溶解)40μl、p-ニトロフェニル-α-D-グルコピラノシド溶液(0.7mM;50mMリン酸カリウム緩衝液(pH7.0)、100mM NaClに溶解)950μlを混和し、37℃、15分間インキュベートした。その後、反応停止液(0.5Mトリス溶液)1mlを添加し、400nmで吸光度を測定した。別に、対照として試料溶液の代わりに蒸留水を、またブランクとしてp-ニトロフェニル-α-D-グルコピラノシド溶液を添加する前に反応停止液を加え同様に操作して吸光度を測定した。対照として、α-グルコシダーゼ阻害作用を持つ事が知られている(+)-カテキンを試験に用いた。
本実施例のα-グルコシダーゼ阻害活性の算出は、本実施例の測定結果を基に、上式(数1)を用いて行った。試料の濃度に対して対数プロットをとりIC50(酵素活性を50%阻害するエキス反応終濃度)を求めた。
本品は、口当たり、風味とも良好なキャンディである。上記のキャンディを高血糖者10名がパネルとなり、上記のキャンディを毎食後に摂取したところ、内4名の血糖値の上昇を抑制した。
また、本実施例のオオウメガサソウ抽出物(実施例1)の代わりに、実施例2のオオウメガサソウ抽出物を使用した以外は同一の方法で製造したキャンディについて、本実施例とはパネル対象者を変更し、本実施例と同一方法でパネル試験を行った結果、本実施例のパネル試験結果と同様に血糖値の上昇抑制などの改善効果を奏する結果を得た。
上記化粧水をパネル10名が1ヶ月連続使用したところ、内6名がアクネやふけが減少した。
また、本実施例のオオウメガサソウ抽出物(実施例2)の代わりに、実施例1のオオウメガサソウ抽出物を使用した以外は同一の方法で製造した化粧水について、本実施例とはパネル対象者を変更し、本実施例と同一方法でパネル試験を行った結果、本実施例のパネル試験結果と同様にアクネやふけの抑制などの改善効果を奏する結果を得た。
Claims (2)
- オオウメガサソウ抽出物を含有するリパーゼ阻害剤。
- オオウメガサソウ抽出物が、オオウメガサソウの水、低級アルコール及び多価アルコールから選ばれる単独又は2種以上の混合溶媒による抽出物である請求項1記載のリパーゼ阻害剤。
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US9072717B2 (en) * | 2007-09-18 | 2015-07-07 | Elc Management Llc | Cosmetic compositions containing alpha glucosidase inhibitors and methods of use |
JP2014177454A (ja) * | 2013-02-18 | 2014-09-25 | Kagoshima Univ | ねじめびわ茶抽出物を含有する飲食品及び医薬品 |
Citations (4)
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JPS6357526A (ja) * | 1986-08-28 | 1988-03-12 | Nippon Shoji Kk | 前立腺肥大症の治療および予防剤 |
JPH04279511A (ja) * | 1991-03-05 | 1992-10-05 | Nonogawa Shoji Kk | 化粧料 |
JPH11335233A (ja) * | 1998-05-22 | 1999-12-07 | Ichimaru Pharcos Co Ltd | メラニン生成抑制剤及び化粧料組成物 |
JP2002241299A (ja) * | 2001-02-13 | 2002-08-28 | Ichimaru Pharcos Co Ltd | メイラード反応修復剤 |
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JPS6357526A (ja) * | 1986-08-28 | 1988-03-12 | Nippon Shoji Kk | 前立腺肥大症の治療および予防剤 |
JPH04279511A (ja) * | 1991-03-05 | 1992-10-05 | Nonogawa Shoji Kk | 化粧料 |
JPH11335233A (ja) * | 1998-05-22 | 1999-12-07 | Ichimaru Pharcos Co Ltd | メラニン生成抑制剤及び化粧料組成物 |
JP2002241299A (ja) * | 2001-02-13 | 2002-08-28 | Ichimaru Pharcos Co Ltd | メイラード反応修復剤 |
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