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ノックアウト漢方の創製とそのエビデンスに関する研究

Research Project

Project/Area Number 19659177
Research Category

Grant-in-Aid for Exploratory Research

Allocation TypeSingle-year Grants
Research Field General internal medicine (including Psychosomatic medicine)
Research InstitutionNagasaki International University

Principal Investigator

正山 征洋  Nagasaki International University, 薬学部, 教授 (70037604)

Co-Investigator(Kenkyū-buntansha) 森永 紀  長崎国際大学, 薬学部, 講師 (60465771)
宇都 拓洋  長崎国際大学, 薬学部, 助手 (90469396)
Project Period (FY) 2007 – 2008
Project Status Completed (Fiscal Year 2008)
Budget Amount *help
¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2008: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2007: ¥2,000,000 (Direct Cost: ¥2,000,000)
Keywordsノックアウト漢方 / 活性成分 / アフィニテイカラム / エビデンス / 生薬 / モノクローナル抗体 / 小型化抗体 / ノックアウトエキス
Research Abstract

芍薬のペオニフロリンと甘草のグリチルリチンに対するscFVを作成しそれらの性状をそれぞれのMAbと比較した。その結果ペオニフロリンに対するscFVを抗ペオニフロリンMAbと比較した結果、両者とも幅広いクロスリアクションを持つ抗体で、ほぼ同一の挙動を示すことが明確となった。抗グリチルリチンscFVは直接的ELISAでは親和性を持つものの、競合的ELISAではグリチルリチンの測定が出来ないことが明らかとなった。
これらの結果からscFVを装着カラムでは十分なアフィニテイを持つカラムは得られないことが明らかになったので、抗グリチルリチンMAbと抗ペオニフロリンMAb装着カラムを作成した。本カラムを用いて、芍薬甘草湯エキスをチャージし、各種溶出溶媒を検討した結果、芍薬甘草湯エキスからグリチルリチンとペオニフロリン両化合物のみを除去し、他の成分は全て保有するエキスの作成に成功した。このことは我々が新たに開発した「イースタンブロッテイング法」と通常の薄層クロマトにより確認し、「ノックアウト漢方エキス」であることを明確にした。薬理効果については目下検討中である。一方、アメリカ人参エキスからジンセノシドReのみを除去したノックアウトエキスは粗エキスに比べ明らかにマウスの体重を上昇させることから、アメリカ人参エキスの体重をコントロールした活性本体はジンセノシドReであることを明らかにした。本研究により創製した「ノックアウト漢方エキス」は活性本体を見出す有効な手段であることを明確にした。

Report

(2 results)
  • 2008 Annual Research Report
  • 2007 Annual Research Report
  • Research Products

    (33 results)

All 2009 2008 2007

All Journal Article (15 results) (of which Peer Reviewed: 12 results) Presentation (10 results) Book (8 results)

  • [Journal Article] Quantitative analysis of ginsenosides Rb1, Rg1, and Re in American ginseng berry and flower samples by ELISA using monoclonal antibodies2009

    • Author(s)
      B. Sritularak, O. Morinaga, CS. Yuan, Y. Shoyama, H. Tanaka
    • Journal Title

      J. Nat. Med. (In press)

    • NAID

      10025826698

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Enzyme-linked immunosorbent assay for total sennosides using anti-sennoside A and B monoclonal antibodies2009

    • Author(s)
      K. Kido, K. Edakuni, O. Morinaga, H. Tanakaa, Y. Shoyama
    • Journal Title

      Fitoterapia 80

      Pages: 28-31

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Development of eastern blotting technique for sennoside A and B using anti-sennoside A and B monoclonal antibodies2009

    • Author(s)
      O. Morinaga, T. Uto, S. Sakamoto, W. Putalun, et al.
    • Journal Title

      Phytochem. Anal. 20

      Pages: 154-158

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Characterization of olivetol synthase, a polyketide synthase putatively involved in cannabinoid biosynthetic pathway2009

    • Author(s)
      F. Taura, S. Tanaka, C. Taguchi, T. Fukamizu, et al.
    • Journal Title

      FEBS Lett. 583

      Pages: 2061-2066

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] An enzyme-linked immunosorbent assay for aconitine-type alkaloids using an anti-aconitine monoclonal antibody2008

    • Author(s)
      K. Kido, K. Edakuni, O. Morinaga, H. Tanakaa, Y. Shoyama
    • Journal Title

      Analytica Chimica Acta 616

      Pages: 109-114

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Quick analysis of baicalin in Scutellariae Radix by enzyme-linked immunosorbent assay using a monoclonal antibody2008

    • Author(s)
      K. Kido, O. Morinaga, Y. Shoyama, H. Tanaka
    • Journal Title

      Talanta 77

      Pages: 346-350

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Pharmacologically active compounds in Anoectochilus and Goodyera species2008

    • Author(s)
      X. M. DU, N. Irino, N. Hurusyo, J. Hayashi, et al.
    • Journal Title

      J. Nat. Med. 62

      Pages: 132-148

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Artemisinin production by shoot regeneration of Artemisia annua L. using thidiazuron2008

    • Author(s)
      W. Lualon, W. De-Eknamkul, H. Tanaka, Y. Shoyama, et al.
    • Journal Title

      Z. Naturforsch. 63c

      Pages: 96-100

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Pharmacologically active saikosaponin in Bupleurum falcatum detected by competitive ELISA and eastern blotting using MAb2008

    • Author(s)
      TK. Shon, S. Zhu, SC. Lee, . Y. Shoyama, H. Tanaka
    • Journal Title

      Plant Prod. Sci. 11

      Pages: 192-197

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Production, characterization of a monoclonal antibody against aristolochie acid-II and development of its assay system2008

    • Author(s)
      M. Tian, H. Tanaka, M. Y. Shang, S. Karashima, et al.
    • Journal Title

      Am. J. Chin. Med. 36

      Pages: 425-436

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Micropropagation of Anoectochilus formosanus Hayata in vitro and pharmacological and chemical investigations2008

    • Author(s)
      X. M. DU, N. Irino, T. Uto, O. Morinaga, Y. Shoyama, et al
    • Journal Title

      Current Topics in Phytochem. 9

      Pages: 79-87

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Crystallization and preliminary X-ray diffraction studies of polyketide synthase-1 (PKS-l)from Cannabis sativa2008

    • Author(s)
      C. Taguchi, F. Taura, T. Tamada, Y. Shoyama, et al.
    • Journal Title

      Acta Crystallographica Section F. 64

      Pages: 217-220

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Research Signpost2007

    • Author(s)
      Z.Lu, T.Masaki, Y.Shoyama, H.Tanaka
    • Journal Title

      Analysis of natural glycosides "Constructiong and expression of a single chain Fv fragment against paeoniflorin in Escherichia coli, and its potential use in and enzyme-linked immunosorbent assay"

      Pages: 14-14

    • Related Report
      2007 Annual Research Report
  • [Journal Article] Research Signpost2007

    • Author(s)
      N.Fukuda, H.Tanaka, Y.Shoyama
    • Journal Title

      Analysis of natural glycosides "Immunoaffinity concentration, one step purification, and preparation of knockout extract"

      Pages: 10-10

    • Related Report
      2007 Annual Research Report
  • [Journal Article] Springer2007

    • Author(s)
      W.Putalun, H.Tanaka, Y.Shoyama
    • Journal Title

      Applications of plant metabolic engineering "Single-chain Fv antibody stimulates biosynthesis of secondary metabolites in plants"

      Pages: 12-12

    • Related Report
      2007 Annual Research Report
  • [Presentation] Functional analysis of licorice extract using knockout extract prepared by anti- glycyrrhizin MAb2009

    • Author(s)
      Y. Shoyama, T. Uto, O. Morinaga
    • Organizer
      The second Taiwan-Japan Chinese Medicine Mini Symposium
    • Place of Presentation
      Taipei, Taiwan
    • Year and Date
      2009-03-04
    • Related Report
      2008 Annual Research Report
  • [Presentation] Pharmacologically activity of crocin in saffron2008

    • Author(s)
      Y. Shoyama
    • Organizer
      2008' Shanghai International Symposium for Pharmaceutical Sciences
    • Place of Presentation
      Shanghai医薬大学
    • Year and Date
      2008-12-19
    • Related Report
      2008 Annual Research Report
  • [Presentation] Preparation of a novel monoclonal antibody against antimalaria drug, artemisinin2008

    • Author(s)
      W. Putalun, H. Tanaka, Y. Shoyama
    • Organizer
      2008' Shanghai International Symposium for Pharmaceutical Sciences
    • Place of Presentation
      Shanghai医薬大学
    • Year and Date
      2008-12-19
    • Related Report
      2008 Annual Research Report
  • [Presentation] Anti-sennoside A and sennoside B monoclonal antiboties2008

    • Author(s)
      O. Morinaga, H. Tanaka, Y. Shoyama
    • Organizer
      2008' Shanghai International Symposium for Pharmaceutical Sciences
    • Place of Presentation
      Shanghai医薬大学
    • Year and Date
      2008-12-19
    • Related Report
      2008 Annual Research Report
  • [Presentation] Production of tetrahydrocannabinolic acid by the biosynthetic enzyme secreted from transgenic Pichia pastoris2008

    • Author(s)
      F. Taura, S. Morimoto, Y. Shoyama
    • Organizer
      2008' Shanghai International Symposium for Pharmaceutical Sciences
    • Place of Presentation
      Shanghai医薬大学
    • Year and Date
      2008-12-19
    • Related Report
      2008 Annual Research Report
  • [Presentation] Crocin, purified from Crocus sativus L., increased non-rapid eye movement sleep in mice2008

    • Author(s)
      K. Aritake, Y. Urade, Y. Shoyama
    • Organizer
      2008' Shanghai International Symposium for Pharmaceutical Sciences
    • Place of Presentation
      Shanghai医薬大学
    • Year and Date
      2008-12-19
    • Related Report
      2008 Annual Research Report
  • [Presentation] Synergistic suppression of inducible nitric oxide synthase expression by glycyrrhizin in lipopolysaccharide-stimulated murine macrophages2008

    • Author(s)
      T. Uto, O. Morinaga, Y. Shoyama
    • Organizer
      2008' Shanghai International Symposium for Pharmaceutical Sciences
    • Place of Presentation
      Shanghai医薬大学
    • Year and Date
      2008-12-19
    • Related Report
      2008 Annual Research Report
  • [Presentation] Construction and expression of specificity-improved single-chain variable fragments against the bioactive naphthoquinone, plumbagin2008

    • Author(s)
      S. Sakamoto, W. Putalun, H. Tanaka, Y. Shoyama
    • Organizer
      2008' Shanghai International Symposium for Pharmaceutical Sciences
    • Place of Presentation
      Shanghai医薬大学
    • Year and Date
      2008-12-19
    • Related Report
      2008 Annual Research Report
  • [Presentation] Recent topics in the fields of pharmaceutical science and pharmacognosy2008

    • Author(s)
      Y. Shoyama
    • Organizer
      第6回モンゴルー日本ジョイントシンポジュウム
    • Place of Presentation
      国立モンゴル健康科学大学
    • Year and Date
      2008-08-19
    • Related Report
      2008 Annual Research Report
  • [Presentation] Application of Monoclonal Antibody against Natural Medicines2008

    • Author(s)
      Y. Shoyama
    • Organizer
      4th Korea-Japan-China Joint Symposium on Pharmacognosy
    • Place of Presentation
      Gangneung, Korea
    • Related Report
      2008 Annual Research Report
  • [Book] 現代医療における漢方薬(教科書)2008

    • Author(s)
      本多義昭, 正山征洋, 荻原幸夫, 竹田忠紘編集
    • Total Pages
      153
    • Publisher
      南江堂
    • Related Report
      2008 Annual Research Report
  • [Book] 大麻研究2008

    • Author(s)
      正山征洋
    • Total Pages
      14
    • Publisher
      長崎国際大学論叢
    • Related Report
      2008 Annual Research Report
  • [Book] 福岡の薬草手帳2008

    • Author(s)
      正山征洋監修
    • Total Pages
      205
    • Publisher
      財団法人福岡市薬剤師会
    • Related Report
      2008 Annual Research Report
  • [Book] 薬用人参のイースタンブロッテイング2008

    • Author(s)
      森永紀, 正山征洋
    • Total Pages
      10
    • Publisher
      長崎国際大学論叢
    • Related Report
      2008 Annual Research Report
  • [Book] 金線蓮の薬理活性成分2008

    • Author(s)
      杜暁鳴, 正山征洋, 古庄憲浩, 林純
    • Total Pages
      3
    • Publisher
      FFIジャーナル
    • Related Report
      2008 Annual Research Report
  • [Book] 薬の種類と漢方薬・生薬を取り巻く最近の変革2008

    • Author(s)
      正山征洋
    • Total Pages
      4
    • Publisher
      人間・植物関係学会誌
    • Related Report
      2008 Annual Research Report
  • [Book] 世界の薬用植物事情-砂漠化と甘草2008

    • Author(s)
      正山征洋
    • Total Pages
      5
    • Publisher
      薬用植物研究
    • Related Report
      2008 Annual Research Report
  • [Book] モンゴルの薬用資源2008

    • Total Pages
      5
    • Publisher
      薬用植物研究
    • Related Report
      2008 Annual Research Report

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Published: 2007-04-01   Modified: 2016-04-21  

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