JP6871760B2 - 抗ウイルス作用促進用組成物 - Google Patents
抗ウイルス作用促進用組成物 Download PDFInfo
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- JP6871760B2 JP6871760B2 JP2017040152A JP2017040152A JP6871760B2 JP 6871760 B2 JP6871760 B2 JP 6871760B2 JP 2017040152 A JP2017040152 A JP 2017040152A JP 2017040152 A JP2017040152 A JP 2017040152A JP 6871760 B2 JP6871760 B2 JP 6871760B2
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Images
Description
(1)チロシンをC末端に有するアミノ酸残基数が2〜10のペプチド若しくはその誘導体又はその薬理学的に許容される塩を有効成分とする抗ウイルス作用促進用組成物。
(2)疎水性アミノ酸−チロシンの配列をC末端に有するペプチド若しくはその誘導体又はその薬理学的に許容される塩を有効成分とする抗ウイルス作用促進用組成物。
(3)疎水性アミノ酸−チロシンの配列を有するジペプチド若しくはその誘導体又はその薬理学的に許容される塩を有効成分とする抗ウイルス作用促進用組成物。
(4)疎水性アミノ酸がバリン、イソロイシン又はフェニルアラニンである(2)又は(3)に記載の抗ウイルス作用促進用組成物。
(5)わかめ由来のペプチドを有効成分とする抗ウイルス作用促進用組成物。
(6)前記ウイルスが、インフルエンザウイルス又は単純ヘルペスウイルスである前記(1)〜(5)のいずれかに記載の抗ウイルス作用促進用組成物。
本発明の抗ウイルス作用促進用組成物は、日常的に長期間にわたり摂取しても安全性に全く問題は無い。
本発明のペプチドがC末端以外にカルボキシル基またはカルボキシレートを有している場合、それらの基がアミド化またはエステル化されているものも本発明のペプチドの誘導体に含まれる。
[ジペプチドの抗ウイルス活性試験]
(1)供試ジペプチド
1)Val−Tyr(商品名:Val−Tyr;SigmaAldrich社製)
2)Ile−Tyr(商品名:H−Ile−Tyr−OH;BACHEM社製)
3)Phe−Tyr(商品名:H−Phe−Tyr−OH;BACHEM社製)
インフルエンザウイルス感染マウスを用い、ジペプチドの抗インフルエンザウイルス活性について調べた。BALB/cマウス(6週齢、雌)を1群当たり10例用い、下記の5群に分けた。各群のマウスにA型インフルエンザウイルス(A/NWS/33、H1N1亜型)を麻酔下で1回の経鼻接種により感染させた。1回の接種量は、2×104PFU(プラーク形成単位)/50μL/マウスとした。各試料をウイルス感染の1週間前から感染後1週間までの計14日間、1日2回(午前9時及び午後6時)経口投与した。尚、タミフルの経口投与は、タミフルを0.05質量%含む生理食塩水を調製して行った。また、ジペプチドの経口投与は、Val−Tyrを0.25質量%含む生理食塩水(第3群)、Ile−Tyrを0.25質量%含む生理食塩水(第4群)、Phe−Tyrを0.25質量%含む生理食塩水(第5群)を各々調製して行った。
第1群:対照(蒸留水) 0.4mL/マウス/day
第2群:タミフル(リン酸オセルタミビル) 0.2mg/マウス/day
第3群:Val−Tyr 1mg/マウス/day
第4群:Ile−Tyr 1mg/マウス/day
第5群:Phe−Tyr 1mg/マウス/day
性器ヘルペスウイルスである単純ヘルペスウイルス2型(HSV−2)の感染マウスを用い、ジペプチドの抗単純ヘルペスウイルス活性について調べた。BALB/cマウス(6週齢、雌)を1群当たり5例用い、下記の5群に分けた。各群のマウスに単純ヘルペスウイルス2型(HSV−2 UW268株)を1回、局所に接種し、感染させた。1回の接種量は、1×103PFU(プラーク形成単位)/20μL/マウスとした。ウイルス接種の6日前及び1日前に、それぞれ、メドロキシプロゲステロン酢酸エステル(medroxyprogesterone 17−acetate)(3mg/マウス)をマウスに皮下注射した。各試料をウイルス感染の1週間前から感染後1週間までの計14日間、1日2回(午前9時及び午後6時)経口投与した。尚、アシクロビルの経口投与は、アシクロビルを0.25質量%含む生理食塩水を調製して行った。また、ジペプチドの経口投与は、Val−Tyrを0.05質量%含む生理食塩水(第3群)、Val−Tyrを0.25質量%含む生理食塩水(第4群)、Ile−Tyrを0.05質量%含む生理食塩水(第5群)、Ile−Tyrを0.25質量%含む生理食塩水(第6群)、Phe−Tyrを0.05質量%含む生理食塩水(第7群)、Phe−Tyrを0.25質量%含む生理食塩水(第8群)を各々調製して行った。
第1群:対照(蒸留水) 0.4mL/マウス/day
第2群:アシクロビル 1mg/マウス/day
第3群:Val−Tyr 0.2mg/マウス/day
第4群:Val−Tyr 1mg/マウス/day
第5群:Ile−Tyr 0.2mg/マウス/day
第6群:Ile−Tyr 1mg/マウス/day
第7群:Phe−Tyr 0.2mg/マウス/day
第8群:Phe−Tyr 1mg/マウス/day
1:腫脹あり
2:腫脹及び発赤あり
3:液滲出あり
4:後ろ足麻痺
5:死亡
[わかめ由来ペプチドの抗ウイルス活性試験]
乾燥わかめ100gに2Lの0.1Mクエン酸緩衝液を加えpHを7.0に調整し、これにアルギン酸リアーゼ(商品名:アルギン酸リアーゼS;ナガセケムテックス社製)100Uを加えた後、45℃で3時間処理した。その後、得られた処理物を3000×gで5分間遠心分離し、上清を廃棄し沈殿物を回収し、わかめから分離された蛋白質を含有する組成物を35g得た。
次に、上記組成物に対し、1500gの蒸留水を加え、ホモジナイズした後、10000Uのプロテアーゼ(商品名:プロテアーゼS「アマノ」;天野エンザイム社製)を加え、pHを8.0に調製した後、72℃にて18時間処理した。その後3000×gで5分間、遠心分離し上清を回収した。回収した上清は、限外濾過膜(商品名:FB02−FC−FUS0181;ダイゼン・メンブレン・システムズ社製)で限外濾過し、透過液を回収した後、凍結乾燥機(型式:RLE II−103;共和真空社製)を用いて、−40℃で2時間予備凍結した後、真空度4Paの条件下、棚温40℃で約12時間かけて凍結乾燥した。得られた凍結乾燥物を0.2mmのスクリーンを用いて粉砕し、粉末状のわかめ由来ペプチド10gを得た。
インフルエンザウイルス感染マウスを用い、わかめ由来ペプチドの抗インフルエンザウイルス活性について調べた。BALB/cマウス(6週齢、雌)を1群当たり10例用い、下記の5群に分けた。各群のマウスにA型インフルエンザウイルス(A/NWS/33、H1N1亜型)を麻酔下で1回の経鼻接種により感染させた。1回の接種量は、2×104PFU(プラーク形成単位)/50μL/マウスとした。各試料をウイルス感染の1週間前から感染後1週間までの計14日間、1日2回(午前9時及び午後6時)経口投与した。尚、タミフルの経口投与は、タミフルを0.05質量%含む生理食塩水を調製して行った。また、わかめ由来ペプチドの経口投与は、わかめ由来ペプチドを2.5質量%含む生理食塩水(第3群)、わかめ由来ペプチドを1.25質量%含む生理食塩水(第4群)、わかめ由来ペプチドを0.25質量%含む生理食塩水(第5群)を各々調製して行った。
第1群:対照(蒸留水) 0.4mL/マウス/day
第2群:タミフル(リン酸オセルタミビル) 0.2mg/マウス/day
第3群:わかめ由来ペプチド 10mg/マウス/day
第4群:わかめ由来ペプチド 5mg/マウス/day
第5群:わかめ由来ペプチド 1mg/マウス/day
性器ヘルペスウイルスである単純ヘルペスウイルス2型(HSV−2)の感染マウスを用い、わかめ由来ペプチドの抗単純ヘルペスウイルス活性について調べた。BALB/cマウス(6週齢、雌)を1群当たり5例用い、下記の5群に分けた。各群のマウスに単純ヘルペスウイルス2型(HSV−2 UW268株)を1回、局所に接種し、感染させた。1回の接種量は、1×103PFU(プラーク形成単位)/20μL/マウスとした。ウイルス接種の6日前及び1日前に、それぞれ、medroxyprogesterone 17−acetate(3mg/マウス)をマウスに皮下注射した。各試料をウイルス感染の1週間前から感染後1週間までの計14日間、1日2回(午前9時及び午後6時)経口投与した。尚、アシクロビルの経口投与は、アシクロビルを0.25質量%含む生理食塩水を調製して行った。また、わかめ由来ペプチドの経口投与は、わかめ由来ペプチドを2.5質量%含む生理食塩水(第3群)、わかめ由来ペプチドを1.25質量%含む生理食塩水(第4群)、わかめ由来ペプチドを0.25質量%含む生理食塩水(第5群)を各々調製して行った。
第1群:対照(蒸留水) 0.4mL/マウス/day
第2群:アシクロビル 1mg/マウス/day
第3群:わかめ由来ペプチド 10mg/マウス/day
第4群:わかめ由来ペプチド 5mg/マウス/day
第5群:わかめ由来ペプチド 1mg/マウス/day
1:腫脹あり
2:腫脹及び発赤あり
3:液滲出あり
4:後ろ足麻痺
5:死亡
Claims (2)
- バリン−チロシン、イソロイシン−チロシン又はフェニルアラニン−チロシンの配列を有するジペプチド又はその薬理学的に許容される塩を有効成分とする抗インフルエンザウイルス作用又は抗単純ヘルペスウイルス作用促進用組成物。
- わかめ由来のペプチドを有効成分とし、わかめ由来のペプチドがバリン−チロシン、イソロイシン−チロシン又はフェニルアラニン−チロシンの配列を有するジペプチド又はその薬理学的に許容される塩を含有する、抗インフルエンザウイルス作用又は抗単純ヘルペスウイルス作用促進用組成物。
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