JP2019156786A - エクソソーム産生促進剤 - Google Patents
エクソソーム産生促進剤 Download PDFInfo
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- JP2019156786A JP2019156786A JP2018047738A JP2018047738A JP2019156786A JP 2019156786 A JP2019156786 A JP 2019156786A JP 2018047738 A JP2018047738 A JP 2018047738A JP 2018047738 A JP2018047738 A JP 2018047738A JP 2019156786 A JP2019156786 A JP 2019156786A
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- exosome production
- exosome
- trans
- production promoter
- sphingadienin
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Abstract
Description
項1. スフィンゴイド塩基を有効成分とするエクソソーム産生促進剤。
項2. スフィンゴイド塩基が、4−トランス,8−トランス−スフィンガジエニン、4−トランス,8−シス−スフィンガジエニン、及びスフィンゴシンよりなる群から選択される少なくとも1種である、項1に記載のエクソソーム産生促進剤。
項3. アミロイドβタンパク質の蓄積に起因する疾患を予防又は改善するために使用される、項1又は2に記載のエクソソーム産生促進剤。
項4. アミロイドβタンパク質の蓄積に起因する認知機能の低下を予防又は改善するために使用される、項1又は2に記載のエクソソーム産生促進剤。
項5. アミロイドβタンパク質の蓄積に起因する記憶障害を予防又は改善するために使用される、項1又は2に記載のエクソソーム産生促進剤。
項6. エクソソーム産生促進用飲食品である、項1〜5のいずれかに記載のエクソソーム産生促進剤。
項7. エクソソーム産生促進用医薬品である、項1〜5のいずれかに記載のエクソソーム産生促進剤。
本発明のエクソソーム産生促進剤は、スフィンゴイド塩基を有効成分として使用する。スフィンゴイド塩基は、スフィンゴ脂質を構成する塩基部分(長鎖アミノアルコール)として公知の化合物である。
本発明のエクソソーム産生促進剤は、前述したスフィンゴイド塩基以外に、本発明の効果を損なわない範囲で、剤型に応じて、他の添加成分を含有してもよい。本発明のエクソソーム産生促進剤に含有され得る添加成分としては、例えば、水、油脂類、ロウ類、炭化水素類、脂肪酸類、高級アルコール類、エステル類、植物抽出エキス類、彗星高分子、界面活性剤、金属石鹸、アルコール類、多価アルコール、pH調製剤、酸化防止剤、紫外線吸収剤、防腐剤、香料、粉黛、増粘剤、色素、キレート剤等が挙げられる。これらの添加成分は、1種類単独で混合しても、2種類以上を組み合わせて使用してもよい。また、これらの添加成分の含量については、使用する添加成分の種類や本発明のエクソソーム産生促進剤の剤型等に応じて適宜設定される。
本発明のエクソソーム産生促進剤の剤型については、特に限定されず、固体状、半固体状、又は液体状のいずれであってもよく、エクソソーム産生促進剤の種類、用途、投与方法などに応じて適宜設定すればよい。
1.スフィンゴイド塩基の調製
こんにゃく抽出物(株式会社ダイセル提供)を10倍量のアセトンで処理し、単純脂質を除去した後、適量のクロロホルム/メタノール/0.4M KOH水溶液(クロロホルム:メタノール:0.4M KOH水溶液の容積比1:1:1)の混合溶媒中でグリセロ脂質をケン化分解し、シリカゲルカラムクロマトグラフィーによりグルコシルセラミドを精製した。精製したグルコシルセラミドを適量のジオキサン/10%水酸化バリウム水溶液(ジオキサン:10%水酸化バリウム水溶液の容積比1:1)中で110℃で加熱分解し、スフィンゴイド塩基を遊離させた。スフィンゴイド塩基は最終的にODSカラムを装備したHPLCで精製し、スフィンゴシン、4−トランス,8−シス−スフィンガジエニン、及び4−トランス,8−トランス−スフィンガジエニンをそれぞれ得た。
ヒト神経芽細胞腫SH-SY5Y細胞(ATCCから購入)を使用し、実験を行った。細胞を、培地(50%Ham’s F12/50%E−MEM、10%仔牛血清含有)と共に6ウェルプレートに2.2×107cells/wellとなるよう播種し、37℃で24時間培養した後、スフィンゴシン(Avanti Polar Lipids社製)、4−トランス,8−シス−スフィンガジエニン、及び4−トランス,8−トランス−スフィンガジエニンを、3%ウシ血清アルブミン(BSA)を含む血清不含培地(50%Ham’s F12/50%E−MEM)に懸濁して、ウェル中での終濃度が5μMとなるように各ウェルに添加した。また、スフィンゴイド塩基の代わりにジメチルスルホキシド(DMSO)をウェル中での終濃度が5μMとなるように各ウェルに添加した場合とコントロールとした。
遠心法で回収したエクソソームを、HEPES/KClバッファーに懸濁した後、ナノパーティクルアナライザーqNano(Izon社)で粒子数を測定した。測定値は、BCA法で算出した由来細胞のタンパク質量(mg)当たりのエクソソーム粒子数粒子数として表示した。エクソソーム粒子数は、測定を2回行って得られた各値の平均値である。また、エクソソーム粒子数は、コントロールを100%とした場合の比率(% of contorol)として算出した。また、得られた値について、One−way ANOVA法による有意差検定(***P<0.001、**P<0.01、*P<0.05)を行った。
結果を図1及び2に示す。図1には、スフィンゴシンを添加した場合のエクソソーム粒子数を示し、図2には、4−トランス,8−シス−スフィンガジエニン、及び4−トランス,8−トランス−スフィンガジエニンンを添加した場合のエクソソーム粒子数を示す。
Claims (7)
- スフィンゴイド塩基を有効成分とするエクソソーム産生促進剤。
- スフィンゴイド塩基が、4−トランス,8−トランス−スフィンガジエニン、4−トランス,8−シス−スフィンガジエニン、及びスフィンゴシンよりなる群から選択される少なくとも1種である、請求項1に記載のエクソソーム産生促進剤。
- アミロイドβタンパク質の蓄積に起因する疾患を予防又は改善するために使用される、請求項1又は2に記載のエクソソーム産生促進剤。
- アミロイドβタンパク質の蓄積に起因する認知機能の低下を予防又は改善するために使用される、請求項1又は2に記載のエクソソーム産生促進剤。
- アミロイドβタンパク質の蓄積に起因する記憶障害を予防又は改善するために使用される、請求項1又は2に記載のエクソソーム産生促進剤。
- エクソソーム産生促進用飲食品である、請求項1〜5のいずれかに記載のエクソソーム産生促進剤。
- エクソソーム産生促進用医薬品である、請求項1〜5のいずれかに記載のエクソソーム産生促進剤。
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WO2022259526A1 (ja) | 2021-06-11 | 2022-12-15 | 株式会社フルステム | エクソソーム産生促進剤及びエクソソーム産生促進方法 |
WO2022259525A1 (ja) | 2021-06-11 | 2022-12-15 | 株式会社フルステム | エクソソーム回収方法 |
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KR20220167354A (ko) | 2021-06-11 | 2022-12-20 | 풀스템 코., 엘티디. | 엑소좀 회수 방법 |
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KR20240000582A (ko) | 2021-06-11 | 2024-01-02 | 풀스템 코., 엘티디. | 엑소좀 생산 촉진제 및 엑소좀 생산 촉진 방법 |
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