JP6021005B2 - 抗菌組成物およびその利用 - Google Patents
抗菌組成物およびその利用 Download PDFInfo
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- JP6021005B2 JP6021005B2 JP2012536558A JP2012536558A JP6021005B2 JP 6021005 B2 JP6021005 B2 JP 6021005B2 JP 2012536558 A JP2012536558 A JP 2012536558A JP 2012536558 A JP2012536558 A JP 2012536558A JP 6021005 B2 JP6021005 B2 JP 6021005B2
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
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Description
〔麹菌〕
麹菌は、日本の伝統的な発酵食品である醤油、味噌、清酒、焼酎などの醸造に不可欠である。麹菌とは、麹づくりに用いられるカビの総称である。
後述する実施例に示すように、麹あるいはその抽出物は、歯周病菌に対する増殖阻害活性を示す。すなわち、麹またはその抽出物は、歯周病菌の増殖を阻害する活性を有している。さらに、麹あるいはその抽出物は、カンジダ症の原因菌(Candida arbicans)や胃潰瘍や胃がんのリスクファクターであるピロリ菌(Helicobacter pylori)の増殖を阻害する活性を有している。
後述する実施例に示すように、麹原料またはその抽出物は、トリプシン様活性を示すジンジパイン(gingipain)に対する阻害活性を示す。すなわち、麹原料またはその抽出物は、ジンジパインの活性を阻害する。ジンジパインは、Pgの生産するシステインプロテアーゼであり、ペプチド切断部位特異性の異なるアルギニンの位置で切断するArg−ジンジパイン(Rgp)とリジンの位置で切断するLys−ジンジパイン(Kgp)の2つの酵素がある。
(1)組成物
1つの局面において、本発明は、歯周病を処置するための有効成分を含有する組成物を提供する。本明細書中にて使用される場合、用語「処置」は、症状の軽減、改善または排除が意図され、治療的(発症後)に行われ得るものだけでなく、予防的(発症前)に行われ得るものもまた包含される。「歯周病を処置するための有効成分」は、上述した「歯周病菌に対する活性物質」および「歯周病菌の生成する毒素に対する活性物質」の少なくとも一方が意図され、本明細書中にて「有効成分」と省略され得る。すなわち、本明細書中にて使用される場合、「有効成分」は、麹またはその抽出物であり得、麹原料またはその抽出物でもあり得る。なお、歯周病菌は口臭の原因でもあるので、本明細書中にて使用される場合、「歯周病の処置」は「口臭の予防または改善」を包含し得る。
1つの局面において、本発明はまた、歯周病を処置するための有効成分を備えているキットを提供する。他の局面において、本発明は、カンジダ菌の増殖を抑制するための有効成分を備えているキットを提供する。本明細書中にて使用される場合、用語「キット」は、特定の材料を内包する容器(例えば、ボトル、プレート、チューブ、ディッシュなど)を備えた包装が意図される。好ましくは、上記材料を使用するための指示書を備えている。本明細書中にてキットの局面において使用される場合、「備えた(備えている)」は、キットを構成する個々の容器のいずれかの中に内包されている状態が意図される。また、本発明のキットは、複数の異なる組成物を1つに梱包した包装であってもよく、容器中に内包された溶液形態の組成物を梱包していてもよい。本発明のキットは、異なる2つ以上の物質を同一の容器に混合して備えても別々の容器に備えてもよい。「指示書」は、紙またはその他の媒体に書かれていても印刷されていてもよく、あるいは磁気テープ、コンピューター読み取り可能ディスクまたはテープ、CD-ROMなどのような電子媒体に付されてもよい。本発明のキットは、上述した組成物を構成するためにもちいられてもよく、上述した組成物に含まれる物質を別々に備えていても、上述した組成物とさらなる成分とを別々に備えていてもよい。
本発明はさらに、歯周病を処置するための方法を提供する。本発明の方法は、本発明の有効成分(または該有効成分を含有する組成物)と、歯周病の原因菌とを接触させる工程を包含する。本発明の有効成分と歯周病の原因菌とを接触させる工程は、歯磨き、指でのペーストの塗布、パッチの貼付であっても、トローチ剤またはチュアブル剤等を口に含むことであってもよく、含嗽剤を用いてうがいをする工程であってもよい。なお、本工程において、本発明の有効成分と歯周病の原因菌とを接触させる時間は、採用される手段に応じて適宜設計されるが、10秒間〜10分間の範囲内が好ましく、30秒間〜10分間の範囲内がより好ましく、1分間〜5分間の範囲内がさらに好ましい。
上述したように、本発明は、歯周病菌だけでなく、ピロリ菌やカンジダ菌などにも適用可能である。すなわち、本発明は、ピロリ菌やカンジダ菌の増殖を阻害するための有効成分、該有効成分を含有する組成物、該有効成分を備えているキット、ならびに、該有効成分、該組成物および該キットを用いるピロリ菌やカンジダ菌の増殖阻害方法を提供する。
4種類の麹菌(A. oryzae、A. sojae、A. kawachii、A. awamori)を用いて、米、麦、醤油麹原料(脱脂大豆と小麦の混合物)、大豆、そら豆、ピーナッツを発酵させて麹を製造した。
醤油麹、丸大豆、大豆麹、および、脱脂大豆と小麦との混合物を用いて、A. oryzaeを用いた場合の醤油麹および大豆麹における抗菌活性をさらに調べた。上記混合物は、醤油麹を製造するための原料であり、具体的には、炒った小麦を粉砕したものと、脱脂した大豆に散水してオートクレーブしたものとの混合物である。醤油麹(O−s)は、上記混合物にA. oryzae S-03を接種して得られた麹である。大豆麹(O−d)は、丸大豆に吸水させてオートクレーブしたものにA. oryzae S-03を接種して得られた麹である。これらの麹から2倍量の水で抽出した後に凍結乾燥したものをサンプルとして用いた。丸大豆に吸水させてオートクレーブしただけのものも併せて用いた。
続いて、醤油麹(O−s)の原液200mLを3倍量のクロロホルムで2回抽出した。次いで、得られたクロロホルム相を脱水および濃縮し、クロロホルム画分としてバイオアッセイに供した。同時に得られた水相を酢酸エチルで2回抽出し、得られた酢酸エチル相を脱水および濃縮し、酢酸エチル画分として、残存した水相を水画分としてバイオアッセイに供した。さらに、水相と酢酸エチル相との間に形成された薄層を中間層としてバイオアッセイに供した。
Pgの生産するシステインプロテアーゼであり、トリプシン様活性を示すジンジパイン(gingipain)の作用に対する阻害活性を調べた。麹または発酵原料からの抽出物(4mg/mLまたは10mg/mL)を、Z−His−Glu−Lys−MCA(ペプチド研究所より購入)とともに、反応溶液(50mM酢酸ナトリウム(pH6.8)、100mM NaCl、10mM DTTを含む。)中にて、37℃で2分間インキュベートした。次いで、この反応溶液に100μg/mLのジンジパイン溶液を添加し、37℃で10分間反応させた。この反応によって生成されたアミノクマリンの蛍光を355/460nmにて測定した。なお、MCAは、4−メチル−7−アミノクマリンであり、Z−His−Glu−Lys−MCAはLys−ジンジパインの基質である。その結果、大豆麹およびその抽出物、ならびに醤油麹原料およびその抽出物が、ジンジパインの活性を強く阻害することがわかった(図2)。
麹抽出物の、カンジダ菌(Candida albicans)に対する抗菌活性を調べた。A. sojae No.9を用いて製造した各種麹の水抽出物(100mg/mL)を染み込ませたペーパーディスクを、カンジダ菌(Candida albicans IFO 01385)を播種したプレート上に配置した後に、37℃で24時間培養することによって、各サンプルのカンジダ菌に対する増殖阻害活性を調べた(図3)。なお、脱脂大豆麹を製造する際には、脱脂大豆を120℃で30分間高圧殺菌した後に、麹菌の分生子を接種した。また、各抽出物は、全て100℃で10分間の加熱処理を行ったものを用いた。示されるように、醤油麹原料は抗カンジダ菌活性を有していないが、脱脂大豆麹または醤油麹の抽出物は、高い抗カンジダ菌活性を有していることがわかった。
麹または発酵原料による各抗菌活性を調べた結果を、表1に示す。なお、脱脂大豆麹を製造する際には、脱脂大豆を120℃で30分間高圧殺菌した後に、麹菌の分生子を接種した。また、各抽出物は、全て100℃で10分間の加熱処理を行ったものを用いた。工事の製造には、A. oryzae S-03を用いた。また、表中の「+/−」はジンジパイン阻害活性がわずかに存在したことを示す。
麹菌(A. oryzae S-03)を用いて得られた脱脂大豆麹20gに対して3倍量の蒸留水(60mL)を加え、室温にて攪拌ながら1時間抽出した。その抽出液(30mL)をメンブランフィルター(ポアサイズ0.2μm)を用いて濾過滅菌した。この濾液を凍結乾燥し、3.1gの粉末(脱脂大豆麹エキス)を得た。
公知の手順に従って、下記成分を下記割合(重量%)で混合して口腔用洗浄液(マウスウオッシュ)を調製した:
グリセリン 5.0
プロピレングリコール 3.0
ラウリル硫酸ナトリウム 1.5
ポリオキシエチレン(60)硬化ヒマシ油 0.4
サッカリンナトリウム 0.3
クエン酸ナトリウム 0.15
メチルパラベン(防腐剤) 0.1
香料 適量
蒸留水
合計 100(重量%)。
麹菌(A. oryzae S-03)を用いて得られた脱脂大豆麹1gに対して3倍量の蒸留水または各種果汁(3mL)を加え、室温にて攪拌ながら1時間抽出した。その抽出液を、メンブランフィルターを用いて濾過滅菌したものを、抗菌活性サンプルとして用いた。果汁は市販の濃縮果汁を1倍まで希釈したもの(100%還元)を、pH6.0に調整した後に用いた。また、果汁は蒸留水と比較して粘度が高いので、ポアサイズ0.45μmのメンブランフィルターを用いた。
Claims (10)
- 麹または該麹の抽出物を含有しており、
前記麹は、大豆麹、脱脂大豆麹、麦麹および醤油麹からなる群より選択される麹であり、
前記麹の抽出物は、大豆麹、脱脂大豆麹、麦麹および醤油麹からなる群より選択される麹の抽出物である、Porphyromonas gingivalisの増殖を阻害するための抗菌組成物(ただし、食品を除く。)。 - 麹または該麹の抽出物を含有しており、
前記麹は、大豆麹、脱脂大豆麹、麦麹および醤油麹からなる群より選択される麹であり、
前記麹の抽出物は、大豆麹、脱脂大豆麹、麦麹および醤油麹からなる群より選択される麹の抽出物である、Helicobacter pyloriの増殖を抑制するための抗菌組成物(ただし、食品を除く。)。 - 前記麹の抽出物が水抽出物、あるいは果汁または野菜汁による抽出によって得られた抽出物である、請求項1または2に記載の組成物。
- 前記麹が、A. oryzae、A. sojae、A. kawachiiおよびA. awamoriからなる群より選択される麹菌を含有している麹である、請求項1〜3のいずれか1項に記載の組成物。
- 前記麹が、大豆麹、脱脂大豆麹および醤油麹からなる群より選択され、且つA. oryzaeを含有している麹である、請求項4に記載の組成物。
- 発酵原料または該発酵原料の抽出物をさらに含有しており、
前記発酵原料が、大豆、脱脂大豆、麦および醤油麹用原料からなる群より選択される発酵原料である、請求項1〜5のいずれか1項に記載の組成物。 - 前記発酵原料の抽出物が水抽出物である、請求項6に記載の組成物。
- 口腔内へ適用可能な形態である、請求項1〜7のいずれか1項に記載の組成物。
- 請求項1〜8のいずれか1項に記載の組成物を含んでいる、医薬部外品。
- 請求項1〜8のいずれか1項に記載の組成物を含んでいる、医薬。
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JPS6447794A (en) * | 1987-08-18 | 1989-02-22 | Masahiro Yamamoto | Production of antibacterial substance by aspergillus |
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