JP4411523B2 - 抗ウイルス剤 - Google Patents
抗ウイルス剤 Download PDFInfo
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- JP4411523B2 JP4411523B2 JP2004061033A JP2004061033A JP4411523B2 JP 4411523 B2 JP4411523 B2 JP 4411523B2 JP 2004061033 A JP2004061033 A JP 2004061033A JP 2004061033 A JP2004061033 A JP 2004061033A JP 4411523 B2 JP4411523 B2 JP 4411523B2
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Landscapes
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Description
コッコミクサの乾燥藻体(10g)にイオン交換水(200mL)を加え、還流下で1時間、加熱抽出を行った。抽出液を遠心分離(3000rpm、15min)し、上清を減圧濃縮しコッコミクサ藻体の抽出物(以下、CEという)70mLを得た。
CE-1〜CE-3の各画分を2N−トリフルオロ酢酸(TFA)に溶解し、121°Cで1時間、加水分解した。窒素ブローによりTFAを除去した後、濃縮した。これに、水素化ホウ素ナトリウム(NaBH4)を加え、生成した単糖類をアルジトールに還元し、過剰のNaBH4 を10%AcOH/MeOHに加え分解し、濃縮乾固した。さらに、MeOHを加え、濃縮乾固を5回行うことでホウ酸イオンを除去した。減圧下で乾燥後、無水酢酸を加え、100°Cで2時間、加熱することでアルジトールアセテートを得た。得られたアルジトールアセテートは、ガスクロマトグラフ/質量分析計により分析した。
HSV−1に対する活性は、アフリカミドリザル腎臓由来のVero細胞を宿主細胞として用いた。このVero細胞を48穴プレートに培養し、0.1PFU(プラーク形成単位)/細胞で感染後、この発明の抗ウイルス剤(CEおよびCE-1〜CE-3)の0.8〜500μg/mlの存在下で処理した。24時間後に収穫して、凍結、溶解処理を3回行った。この検体を適宜希釈して、35mmディッシュに別に培養したVero細胞に感染させ、翌日に染色後、プラークを計数した。無添加対照区のプラーク数を100%として、50%ウイルス増殖阻止濃度(IC50)を算出した。
IFVに対する活性は、イヌ腎臓由来のMDCK細胞を宿主細胞として用いた。このMDCK細胞を48穴プレートに培養し、0.1PFU/細胞で感染後、この発明の抗ウイルス剤(CEおよびCE-1〜CE-3)の0.8〜500μg/mlの存在下で処理した。以下、プラークアッセイはHSV−1と同様の方法で行った。
感染力のあるウイルスそのものは用いない細胞間融合アッセイを行った。すなわち、HIVの糖蛋白質であるgp160(gp120/gp41)を発現しているHeLa細胞(gp160+ HeLa細胞)と、宿主側のレセプターであるCD4を発現しているHeLa細胞(CD+ HeLa細胞)とを一定の割合で混合培養することによって、両者の細胞間で融合(多核巨細胞の形成)が起こることを利用して、この融合に対する阻止効果を検討した。この発明の抗ウイルス剤(CEおよびCE-1〜CE-3)の0.8−500μg/mlの存在下で24時間の混合培養を行った後、ギムザ染色し、顕微鏡下で多核巨細胞数を測定した。無添加対照区の巨細胞数を100%として、50%細胞融合阻止濃度(IC50)を算出した。
HCoVに対する活性は、ヒト胎児肺由来のMRC−5細胞を宿主細胞として用いた。このMRC−5細胞を48穴プレートに培養し、0.001TCID50(50%培養細胞感染量)/細胞で感染後、この発明の抗ウイルス剤(CEおよびCE-1〜CE-3)の0.8〜500μg/mlの存在下で処理した。3日後に収穫して、この検体を適宜希釈して、96穴プレートに別に用意したMRC−5細胞に感染させ、5日間培養する。細胞変性効果(CPE)の有無を判定して、Reed−Muench法によって、50%CPE阻止濃度(IC50)を算出した。
なお、以下に示す表の中で、A区はウイルス感染の時から収穫に至るまでの期間中にこの発明の抗ウイルス剤が存在することを、またB区はウイルス感染直後から収穫に至るまでの期間中にこの発明の抗ウイルス剤が存在することをそれぞれ意味する。
Claims (3)
- 緑色植物門(Chlorohta) 、緑藻綱(Chlorophyeae)、クロロコッカム目(Chlorococcales)、クロロコッカム科(Chlorococcaceae) に属するコッコミクサ・ミノール(Coccomyxa minor) 又はコッコミクサ・グロエオボトリディフォルミス(Coccomyxagloeobotrydiformis)としたコッコミクサ藻体の熱水抽出物から得られた多糖体画分を有効成分としてなり、単純ヘルペスウイルス1型、A型インフルエンザウイルス、ヒト免疫不全ウイルス、ヒトコロナウイルスに対する抗ウイルス活性を有することを特徴とする抗ウイルス剤。
- 前記多糖体画分が、前記コッコミクサ藻体の抽出物の蒸留水による溶出画分であることを特徴とする請求項1記載の抗ウイルス剤。
- 前記多糖体画分が、前記コッコミクサ藻体の抽出物の蒸留水による溶出後の無機塩溶液による溶出画分であることを特徴とする請求項1記載の抗ウイルス剤。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
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JP2004061033A JP4411523B2 (ja) | 2004-03-04 | 2004-03-04 | 抗ウイルス剤 |
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JP2004061033A JP4411523B2 (ja) | 2004-03-04 | 2004-03-04 | 抗ウイルス剤 |
Publications (2)
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JP2005247757A JP2005247757A (ja) | 2005-09-15 |
JP4411523B2 true JP4411523B2 (ja) | 2010-02-10 |
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JP2004061033A Expired - Fee Related JP4411523B2 (ja) | 2004-03-04 | 2004-03-04 | 抗ウイルス剤 |
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2023085351A1 (ja) | 2021-11-11 | 2023-05-19 | 株式会社デンソー | T細胞の分化調節剤及び組成物 |
Families Citing this family (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
TWI461437B (zh) * | 2010-04-09 | 2014-11-21 | Univ Nat Taiwan | 快速萃取褐藻糖膠之方法 |
JP5597160B2 (ja) * | 2011-04-15 | 2014-10-01 | 株式会社日健総本社 | 抗ウイルス剤及びその製法 |
JP6125962B2 (ja) * | 2013-09-24 | 2017-05-10 | 株式会社デンソー | 抗ウイルス剤 |
JP2016065037A (ja) * | 2014-09-17 | 2016-04-28 | 株式会社日健総本社 | 抗ウイルス剤の製法及び該製法によって得られた抗ウイルス剤 |
EP4446337A2 (en) * | 2017-09-22 | 2024-10-16 | Centre National de la Recherche Scientifique | Mutated glycoprotein of vesicular stomatitis virus |
JP2020019729A (ja) * | 2018-07-31 | 2020-02-06 | 株式会社デンソー | ヘルペスウイルス回帰発症抑制剤 |
JP7233043B2 (ja) * | 2018-07-31 | 2023-03-06 | 株式会社デンソー | 抗インフルエンザ剤 |
WO2020026953A1 (ja) * | 2018-07-31 | 2020-02-06 | 株式会社デンソー | 抗ヘルペスウイルス剤 |
JP7378091B2 (ja) * | 2018-07-31 | 2023-11-13 | 株式会社デンソー | 殺ウイルス剤 |
GB2579600B (en) * | 2018-12-05 | 2023-07-05 | Byotrol Plc | Anti-viral composition |
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
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WO2023085351A1 (ja) | 2021-11-11 | 2023-05-19 | 株式会社デンソー | T細胞の分化調節剤及び組成物 |
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