JP2017226642A - ジンセノサイドf1を有効成分として含む免疫増強用組成物 - Google Patents
ジンセノサイドf1を有効成分として含む免疫増強用組成物 Download PDFInfo
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- JP2017226642A JP2017226642A JP2017049803A JP2017049803A JP2017226642A JP 2017226642 A JP2017226642 A JP 2017226642A JP 2017049803 A JP2017049803 A JP 2017049803A JP 2017049803 A JP2017049803 A JP 2017049803A JP 2017226642 A JP2017226642 A JP 2017226642A
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- ginsenoside
- cells
- pharmaceutical composition
- natural killer
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Abstract
【解決手段】本発明の免疫増強用組成物は、ジンセノサイドF1を有効成分として含む。
【選択図】図4a
Description
また、本発明はジンセノサイドF1を有効成分として含む免疫増強用食品組成物に関する。
また、本発明はジンセノサイドF1及びナチュラルキラー細胞を含むパーフォリンまたはグランザイム生産用組成物に関する。
本発明の他の目的は、ジンセノサイドF1を有効成分として含む免疫増強用食品組成物を提供することにある。
本発明のもう一つの目的は、ジンセノサイドF1及びナチュラルキラー細胞を含むパーフォリンまたはグランザイム生産用組成物を提供することにある。
本発明における用語、「ジンセノサイドF1」は、高麗人参や紅参などの主要な活性成分であるサポニンの一種であり、PPT(protopanaxatriol)系に属するジンセノサイドを意味する。
前記先天性免疫細胞は、ナチュラルキラー細胞(natural killer cell、NK cell)、免疫細胞、末梢血単核細胞(peripheral blood mononuclear cell、PBMC)及び樹枝状細胞(dendritic cell)を含む群から選択されたいずれか一つであってもよいが、これに限定されるものではなく、具体的にはナチュラルキラー細胞であってもよい。
本発明における用語、「脱顆粒活性」は、分泌顆粒内に蓄積された物質を放出する活性を意味し、本発明で脱顆粒活性は先天性免疫細胞内に蓄積された物質を放出する活性であってもよい。前記先天性免疫細胞の脱顆粒活性は、一般的に脱顆粒活性を評価するために用いられるマーカーであるCD107aの発現を分析して測定することができるが、これに限定されるものではない。
本発明の一実施例では、ジンセノサイドF1をナチュラルキラー細胞に前処理して、前記細胞の細胞傷害活性を評価した結果、標的細胞の細胞溶解の程度が高く表示され(図3a)、特に、1次ナチュラルキラー細胞に前処理した場合も、標的細胞の細胞溶解程度が高く表示されたことを確認した(図4a)。また、ナチュラルキラー細胞に対して感受性であるRMA−S細胞の減少を通じてジンセノサイドF1のin vivo細胞傷害活性の有効性を評価した場合も、RMA−Sの減少効果が優れていることを確認した(図7)。
本発明の一実施例では、ジンセノサイドRg3をナチュラルキラー細胞に前処理して前記細胞の細胞傷害活性を評価した結果、ナチュラルキラー細胞のCD107aの発現が増加し(図2b及び2c)、標的細胞の細胞溶解の程度が高く表示され(図3b)、特に、1次ナチュラルキラー細胞に前処理した場合も、標的細胞の細胞溶解程度が高く示されることを確認した(図4b)。したがって、本発明の薬学組成物は、免疫増強活性を有するジンセノサイドRg3をさらに含むことができることが分かった。
ジンセノサイドF1とジンセノサイドRg3との比率が1:0.1の重量比以下の場合、ジンセノサイドRg3による免疫増強効果がほとんど示されず、1:1の重量比以上である場合、混合による免疫増強の相乗効果があまり示されないことがある。
本発明における用語、「細胞傷害因子」は、細胞傷害活性を示す物質を意味し、本発明において前記細胞傷害因子はナチュラルキラー細胞から分泌され、細胞傷害活性を示す物質であり、具体的にはパーフォリンまたはグランザイムでありうるが、これに限定されるものではない。
本発明において、「ジンセノサイドF1」及び「免疫強化」は、前記で説明した通りである。
前記組成物は、生理学的に許容可能な担体をさらに含むことができるが、担体の種類は特に限定されず、当該技術分野で通常使用される担体であれば、いずれのものであっても使用することができる。
本発明において、「ジンセノサイドF1」及び「免疫強化」は前記で説明した通りである。
前記の「薬学的に有効な量」は、医学的治療に適用可能な合理的な受益/リスク比で疾患を治療するのに十分な量を意味し、有効容量レベルは疾患の重症度、薬物の活性、患者の年齢、体重、健康、性別、患者の薬物に対する敏感度、使用された本発明の組成物の投与時間、投与経路及び排出比率、治療期間、使用された本発明の組成物と配合または同時に使用される薬物を含む要素及びその他の医学分野でよく知られている要素により決定してもよい。例えば、ジンセノサイドF1を含む前記薬学組成物は、1日に0.0001〜1000mg/kgで、具体的には、0.001〜100mg/kgで投与してもよい。
本発明において、「ジンセノサイドF1」及び「ナチュラルキラー細胞」は、前記で説明した通りである。
フローサイトメトリー分析(fluorescence activated cell sorter、FACS)を使用してジンセノサイドF1による先天性免疫細胞の脱顆粒(degranulation)活性の程度を測定した。
ジンセノサイドF1がナチュラルキラー細胞の細胞傷害活性を増強させるかどうかを確認するために、標的細胞の細胞溶解(cytolysis)程度を測定した。
実施例3−1:1次ナチュラルキラー細胞の分離
健康なドナーの血液(50ml)をヘパリンナトリウムを含む真空採血器細胞準備チューブ(Vacutainer Cell Preparation Tube、BD Biosciences)に採取した。その後、前記血液を遠心分離して血小板と白血球に形成された白の薄い層であるバフィーコート(buffy coat)を回収した。前記バフィーコートをPBSで洗浄し、得られた末梢血単核細胞をhuman NK cell negative selection kit(Miltenyl Biotech)を使用して分離した。分離されたナチュラルキラー細胞の一部とFACSバッファとを混合した後、FACS機械及び蛍光色素(fluorochrome)が結合された抗-CD3、抗-CD56及び抗-CD107a抗体を用いて、ナチュラルキラー細胞(CD3-/CD16+)の純度、CD56dim/CD16+、CD56bright/CD16-及びCD56bright/CD16+の数及び割合を測定して、純度が95%以上であるナチュラルキラー細胞を実験に使用した。
前記実施例3−1の方法で分離された1次ナチュラルキラー細胞を用いて前記実施例2と同様の方法でジンセノサイドF1が1次ナチュラルキラー細胞の細胞傷害活性を増強させるかどうかを確認した。
ジンセノサイドF1のナチュラルキラー細胞ベースの免疫増強機序を解明するために、共焦点顕微鏡(confocal microscopy)を用いた生細胞イメージング(live cell imaging)により単一細胞のカルシウム流動分析を行った。
ジンセノサイドF1によるナチュラルキラー細胞の細胞傷害活性機序を分析するために、前記実施例3−1の方法で純粋分離した1次ナチュラルキラー細胞にF1またはRg1を処理した。12時間経過した後、総RNAを分離した後、ナチュラルキラー細胞の代表的な細胞傷害因子であるパーフォリン(perforin)及びグランザイムB(granzyme B)の発現を定量的RT−PCRで分析した。
ジンセノサイドF1のin vivo細胞傷害活性の有効性を評価するために、ナチュラルキラー細胞に対して感受性であるRMA−S(MHC class I-)と抵抗性を有するRMA(MHC class I+)標的細胞システムを使用した。
Claims (12)
- ジンセノサイドF1を有効成分として含む、免疫増強用薬学組成物。
- 前記免疫が先天性免疫である、請求項1に記載の薬学組成物。
- 前記薬学組成物が、ジンセノサイドRg3をさらに含むものである、請求項1に記載の薬学組成物。
- 前記薬学組成物が、ジンセノサイドF1とジンセノサイドRg3とを1:0.1〜1:1の重量比で含むものである、請求項3に記載の薬学組成物。
- 前記薬学組成物が先天性免疫細胞の活性を増進させるものである、請求項1に記載の薬学組成物。
- 前記先天免疫細胞が、ナチュラルキラー細胞(natural killer cell、NK cell)、免疫細胞、末梢血単核細胞(peripheral blood mononuclear cell、PBMC)、及び樹枝状細胞(dendritic cell)を含む群から選択されるいずれか一つのものである、請求項5に記載の薬学組成物。
- 前記薬学組成物が、ナチュラルキラー細胞(natural killer cell、NK cell)の脱顆粒活性を促進させるものである、請求項1に記載の薬学組成物。
- 前記薬学組成物が、ナチュラルキラー細胞(natural killer cell、NK cell)の細胞傷害活性を増進させるものである、請求項1に記載の薬学組成物。
- 前記薬学組成物が、ナチュラルキラー細胞の細胞傷害因子の発現を増加させるものである、請求項1に記載の薬学組成物。
- 前記細胞傷害因子が、パーフォリン(perforin)またはグランザイム(granzyme)であるものである、請求項9に記載の薬学組成物。
- ジンセノサイドF1を有効成分として含む、先天性免疫増強用食品組成物。
- ジンセノサイドF1及びナチュラルキラー細胞を含む、パーフォリンまたはグランザイム生産用組成物。
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CN107519189A (zh) | 2017-12-29 |
US20170368084A1 (en) | 2017-12-28 |
JP6515126B2 (ja) | 2019-05-15 |
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US10682366B2 (en) | 2020-06-16 |
CA2961400A1 (en) | 2017-12-22 |
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