Development of gram-positive bacteria sepsis therapy by inhibition of bacterial proteases.

• Project/Area Number: 18390125
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Experimental pathology
• Research Institution: Kumamoto University
• Principal Investigator: IMAMURA Takahisa Kumamoto University, 大学院・医学薬学研究部, 准教授 (20176499)
• Co-Investigator(Kenkyū-buntansha): 梅山 秀明 北里大学, 薬学部, 教授 (20050619)
• Project Period (FY): 2006 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥10,760,000 (Direct Cost: ¥9,500,000、Indirect Cost: ¥1,260,000); Fiscal Year 2008: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000); Fiscal Year 2007: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000); Fiscal Year 2006: ¥5,300,000 (Direct Cost: ¥5,300,000)
• Keywords: 黄色ブドウ球菌 / プロテアーゼ / 阻害剤 / 血管漏出 / 敗血性ショック / システインプロテアーゼ / 敗血症 / 治療 / ショック / 凝固 / 線溶 / 欠損株

Research Abstract

黄色ブドウ球菌放出病原プロテアーゼstaphopain A とBの立体構造に基づいて活性中心部の鍵穴に当てはまる化学物質を30万種類程度の試薬リストからコンピュータを使ってドッキング計算より選出し、阻害効果を調べた。Staphopain BについてはKi値が数10μM程度の阻害物質しか得られなかったが、Aに対して最も強い阻害効果をもつchemical #11は、Ki値1.3μMで拮抗型阻害を示し、本菌敗血性ショック発症に直結するstaphopain Aの血管漏出作用を20 μM でほぼ完全に抑制した。さらに他の細菌や細胞、血漿由来のプロテアーゼに対してもその種類や基質特異性とは無関係に阻害効果がみられたことから、プロテアーゼが関係する疾患への治療を含めた多方面への応用性が示唆された。

Research Products

• [Journal Article] A fatal hypotension by sildenafil in an end-stage renal disease patient with hypertension and abnormal pharmacokinetics of the medicine (2009)
  • Author(s): Wada, Y., Kikuchi, K., & Imamura, T.
  • Journal Title: Nephrology 14 Pages: 357-358
  • Peer Reviewed
• [Journal Article] Lysine-specific gingipain promotes lipopolysaccharides- and active vitamin D_3-induced osteoclast differentiation by degrading osteoprotegerin (2009)
  • Author(s): Yasuhara, R., Miyamoto, Y., Takami, M., Imamura, T., Potempa, J., Yoshimura, K., & Kamijo, R.
  • Journal Title: Biochem. J 419 Pages: 159-166
  • Peer Reviewed
• [Journal Article] Lysine-specific gingipain promotes lipopolysaccharides- and active vitamin D_3-induced osteoclast differentiation by degrading osteoprotegerin. (2009)
  • Author(s): Yasuhara, R.
  • Journal Title: Biochem. J. 419 Pages: 159-166
  • Peer Reviewed
• [Journal Article] Thrombin in synovial fluid as a marker of synovial inflammation: a definite measurement by ELISA and correlation with VEGF (2008)
  • Author(s): Kitamoto, Y., Nakamura, E., Kudo, S., Tokunaga, H., Murakami, E., Noguchi, K., & Imamura, T.
  • Journal Title: Clinica Chimica Acta 389 Pages: 159-160
  • Peer Reviewed
• [Journal Article] Prostate expression of endothelins and their receptors in rat growth. Reprod (2008)
  • Author(s): Wada. Y., Takahashi, W., Latifpour, J., Weiss, R. M., Matsumoto, K., Matsui, K., Yamada, G., and *Imamura, T.
  • Journal Title: Fertil. Develop 20 Pages: 750-759
  • Peer Reviewed
• [Journal Article] Production of C5a by ASP, a serine protease released from Aeromonas sobria (2008)
  • Author(s): Nitta, H., Imamura, T., Wada, Y., Irie, A., Kobayashi, H., Okamoto, K., & Baba, H
  • Journal Title: J. Immunol 181 Pages: 3602-3608
  • Peer Reviewed
• [Journal Article] Impaired plasma clotting induction through fibrinogen degradation by ASP, a serine protease released from Aeromonas sobria (2008)
  • Author(s): Imamura, T., Nitta, H., Wada, Y., Kobayashi, H., & Okamoto, K
  • Journal Title: FEMS Mirobiol. Lett 284 Pages: 35-42
  • Peer Reviewed
• [Journal Article] Anti-VEGF therapy for crescentic glomerulonephritis? (2008)
  • Author(s): Kitamoto, Y., Taguma, Y., Arizono, K., & Imamura, T
  • Journal Title: Am. J. Kidney Dis 52 Pages: 710-711
  • Peer Reviewed
• [Journal Article] Coexistence of serus cystadenoma and ductal adenocarcinoma in the pancreas : A case report and literature review (2008)
  • Author(s): Nitta, H., Hirota, M., Ohkado, A., Motomura, Y., Chikamoto, A., Shibata, M., Takamori, H., Kanemitsu, K., Imamura, T., Yamamoto, T., & Baba, H
  • Journal Title: Pancreas 36 Pages: 218-219
  • Peer Reviewed
• [Journal Article] Low serum PSA levels of diabetes mellitus-caused end-stage renal disease patients (2008)
  • Author(s): Wada, Y., Kikuchi, K., Kikkawa, H., & Imamura, T
  • Journal Title: Nephrol. Dial. Transpl. Plus 1 Pages: 58-59
  • Peer Reviewed
• [Journal Article] Modified technique for improving tubeless cutaneous ureterostomy using the Ariyoshi method (2008)
  • Author(s): Wada, Y., Kikuchi, K., Suenaga, T., Imamura, T., Matsumoto, K., & Kodama, K
  • Journal Title: Int. J. Urol 15 Pages: 144-150
  • Peer Reviewed
• [Journal Article] Impaired plasma clotting induction through fibrinogen degradation by ASP, a serine protease released from Aeromonas sobria. (2008)
  • Author(s): Imamura, T.
  • Journal Title: FEMS Mirobiol. Lett. 284 Pages: 35-42
  • Peer Reviewed
• [Journal Article] Production of C5a by ASP, a serine protease released from Aeromonas sobria. (2008)
  • Author(s): Nitta, H.
  • Journal Title: J. Immunol. 181 Pages: 3602-3608
  • Peer Reviewed
• [Journal Article] Activation of prothrombin by ASP, a serine protease released from Aeromonas sobria (2007)
  • Author(s): Nitta, H., Kobayashi, H., Irie, A., Baba, H., Okamoto, K., & Imamura, T
  • Journal Title: FEBS lett 581 Pages: 5935-5939
  • Peer Reviewed
• [Journal Article] Augmentation of vascular permeability of VEGF is enhanced by KDR-binding proteins (2007)
  • Author(s): Yamazaki, Y., Nakano, Y., Imamura, T., & Morita, T
  • Journal Title: Biochem. Biophys. Res. Commun 355 Pages: 693-699
  • Peer Reviewed
• [Journal Article] Induction of vascular leakage through kinin release by a serine proteinase from Aeromonas sobria (2006)
  • Author(s): Imamura, T., Kobayashi, H., Khan, R., Nitta, H., & Okamoto, K
  • Journal Title: J. Immunol 177 Pages: 8723-8729
  • Peer Reviewed
• [Journal Article] Arginine-specific gingipains from Porphyromonas gingivalis deprive protective functions of secretory leukocyte inhibitor in periodontal tissue (2006)
  • Author(s): Into, T., Inomata, M., Kanno, Y., Matsuyama, T., Machigashira, M., Izumi,Y., Imamura, T., Nakashima, M., Noguchi, T., & Matsushita, K
  • Journal Title: Clin. Exp.Immunol 145 Pages: 545-554
  • Peer Reviewed
• [Journal Article] Urokinase immobilization suppresses inflammatory responses to polyurethane tubes implanted in rabbit muscles (2006)
  • Author(s): Lai, Z.-F., Imamura, T., Koike, N., and Kitamoto, Y
  • Journal Title: J. Biomed. Mater. Res 76A Pages: 81-85
  • Peer Reviewed
• [Journal Article] DX-9065a inhibits proinflammatory events induced by gingipains and factor Xa (2006)
  • Author(s): Matsushita, K., Imamura, T., Tomikawa, M., Tancharoen, S., Tatsuyama, S., & Maruyama, I
  • Journal Title: J. Periodont. Res 41 Pages: 148-156
  • Peer Reviewed
• [Journal Article] Selcetive inhibition of Porphyromonas gingivalis growth by a factor Xa inhibitor, DX-9065a (2006)
  • Author(s): 17.Matsushita, K., Imamura, T., Tancharoen, S., Tatsuyama, S., Tomikawa, M.,Travis, J., Potempa, J., Torii, M., & Maruyama, I
  • Journal Title: J. Periodont. Res 41 Pages: 171-176
  • Peer Reviewed
• [Journal Article] Urokinase immobilization suppresses inflammatory responses to polyurethane tubes implanted in rabbit muscles. (2006)
  • Author(s): Lai, Z.-F., Imamura, T., 他2名
  • Journal Title: J.Biomed.Mater.Res. 76A Pages: 81-85
• [Journal Article] DX-9065a inhibits proinflammatory events induced by gingipains and factor Xa. (2006)
  • Author(s): Matsushita, K., Imamura, T., 他4名
  • Journal Title: J.Periodont.Res. 41 Pages: 148-156
• [Journal Article] Selcetive inhibition of Porphyromonas gingivalis growth by a Factor Xa inhibitor, DX-9065a. (2006)
  • Author(s): Matsushita, K., Imamura, T., 他4名
  • Journal Title: J.Periodont.Res. 41 Pages: 171-176
• [Journal Article] Arginine-specific gingipains from Porphyromonas gingivalis deprive protective functions of secretory leukocyte inhibitor in periodontal tissue. (2006)
  • Author(s): Into, T., Imamura, T., 他8名
  • Journal Title: Clin.Exp.Immunol. 145 Pages: 545-554
• [Journal Article] Induction of vascular leakage through kinin release by a serine proteinase from Aeromonas sobria. (2006)
  • Author(s): Imamura, T., Kobayashi, H., 他3名
  • Journal Title: J.Immunol. 117 Pages: 8723-8729
• [Journal Article] Augmentation of vascular permeability of VEGF by KDR-binding proteins. (2006)
  • Author(s): Yamazaki, Y., Imamura, T., 他2名
  • Journal Title: Biochem.Biophys.Res.Commun. 355 Pages: 693-699
• [Presentation] 黄色ブドウ球菌, MRSA 産生protease によるDIC 発症のメカニズム (2008)
  • Author(s): 新田英利
  • Organizer: 第108日本外科学会
  • Place of Presentation: 長崎
  • Year and Date: 2008-05-16
• [Presentation] 黄色ブドウ球菌, MRSA産生proteaseによるDIC発症のメカニズム (2008)
  • Author(s): 新田英利
  • Organizer: 第108日本外科学会
  • Place of Presentation: 長崎市長崎ブリックホール
  • Year and Date: 2008-05-16
• [Presentation] DICの病態把握におけるFDPとD-ダイマーの意味 (2008)
  • Author(s): 内場光浩
  • Organizer: 第55回日本臨床検査医学会総会
  • Place of Presentation: 名古屋
• [Presentation] ALアミロイドーシスにおける過剰線溶と病態形成におけるuPAの関与 (2008)
  • Author(s): 内場光浩
  • Organizer: 第31回日本血栓止血学会学術集会
  • Place of Presentation: 大阪
• [Presentation] Aeromonas sobriaセリンプロテアーゼASPによる血漿凝固促進とトロンビン産生 (2008)
  • Author(s): 今村隆寿
  • Organizer: 第31回日本血栓止血学会学術集会
  • Place of Presentation: 大阪
• [Presentation] ALアミロイドーシスの過剰線溶および病態形成には形質細胞からのuPA が関与している (2008)
  • Author(s): 内場光浩
  • Organizer: 第19回九州血管血栓フォーラム
  • Place of Presentation: 鹿児島
• [Presentation] uPA産生形質細胞により過剰線溶をきたしたAL アミロイドーシス (2008)
  • Author(s): 立津央
  • Organizer: 第90回日本血液学会総会
  • Place of Presentation: 京都
• [Presentation] Aeromonas sobriaセリンプロテアーゼのプロトロンビン活性化による血液凝固誘導作用 (2008)
  • Author(s): 今村隆寿
  • Organizer: 第13回病態と治療におけるプロテアーゼとインヒビター学会
  • Place of Presentation: 大阪
• [Presentation] 黄色ブドウ球菌、MRSA 産生protease による敗血症性ショックおよびDIC発症のメカニズム (2008)
  • Author(s): 新田英利
  • Organizer: 第63回日本消化器外科学会
  • Place of Presentation: 札幌
• [Presentation] Role of uPA in excessive fibrinolysis observed in AL-amyloidosis (2008)
  • Author(s): Uchiba, M.
  • Organizer: XIXth International Congress on Fibrinolysis and Proteolysis
  • Place of Presentation: Viena, Austria
• [Presentation] ハブ毒中で単独で作用する補体活性化因子の構造と機能について (2008)
  • Author(s): 上田直子
  • Organizer: 第52回毒素シンポジウム
  • Place of Presentation: 山梨県山中湖村
• [Presentation] Aeromonas sobria放出セリンプロテアーゼのC5a 産生活性 (2008)
  • Author(s): 今村隆寿
  • Organizer: 第97回日本病理学会総会
  • Place of Presentation: 金沢
• [Presentation] 糖尿病性末期腎不全患者のPSA 低値と前立腺癌低罹患率 : 血液透析患者の原疾患別比較 (2008)
  • Author(s): 和田孝浩
  • Organizer: 第96回日本泌尿器科学会総会
  • Place of Presentation: 横浜
• [Presentation] Aeromonas sobria放出セリンプロテアーゼのC5a産生活性 (2008)
  • Author(s): 今村隆寿
  • Organizer: 第97回日本病理学会総会
  • Place of Presentation: 金沢市石川県立音楽堂
• [Presentation] 黄色ブドウ球菌、MRSA産生proteaseによる敗血症性ショックおよびDIC発症のメカニズム. (2008)
  • Author(s): 新田英利
  • Organizer: 第63回日本消化器外科学会
  • Place of Presentation: 札幌市ロイトン札幌
• [Presentation] Aeromonas sobriaセリンプロテアーゼのプロトロンビン活性化による血液凝固誘導作用 (2008)
  • Author(s): 今村隆寿
  • Organizer: 第13回病態と治療におけるプロテアーゼとインヒビター学会
  • Place of Presentation: 大阪市千里ライフサイエンスセンター
• [Presentation] Aeromonas sobriaセリンプロテアーゼASPによる血漿凝固促進とトロンビン産生 (2008)
  • Author(s): 今村隆寿
  • Organizer: 第31回日本血栓止血学会学術集会
  • Place of Presentation: 大阪市大阪国際交流センター
• [Presentation] 胆嚢炎および胆嚢癌における8-nitroguanine の発現とその意義 (2007)
  • Author(s): 新田英利
  • Organizer: 第107日本外科学会
  • Place of Presentation: 大阪
  • Year and Date: 2007-04-13
• [Presentation] 尿トロンビン測定による腎炎病巣での凝固反応活性化とその制御の解析 (2007)
  • Author(s): 北本康則
  • Organizer: 第7回TTMフォーラム
  • Place of Presentation: 東京
  • Year and Date: 2007-03-10
• [Presentation] ハブ毒中の新規なC3/C5コンベルターゼの精製と緒性質 (2007)
  • Author(s): 鈴木美緒
  • Organizer: 第30回日本分子生物学会年会, 第80回日本生化学会大会合同大会
  • Place of Presentation: 横浜
• [Presentation] 尿トロンビンのATIII難抑制性と腎炎部血管外凝固反応活性化依存性 (2007)
  • Author(s): 北本康則
  • Organizer: 第30回日本血栓止血学会学術集会
  • Place of Presentation: 三重
• [Presentation] Production of C5a by a serine proteinase from Aeromonas sobria, leading to Neutrophil Migration and vascular leakage (2007)
  • Author(s): Imamura, T
  • Organizer: 4th International Symposium of Proteolysis
  • Place of Presentation: Patras, Greece
• [Presentation] Aeromonas sobriaセリンプロテアーゼによるアナフィラトキシンC5aの産生 (2007)
  • Author(s): 今村隆寿
  • Organizer: 第12回病態と治療におけるプロテアーゼとインヒビター学会
  • Place of Presentation: 大阪
• [Presentation] Augmentation of vascular permeability of vascular endothelial growth factor(VEGF)is enhanced by VEGF receptor 2-binding proteins (2007)
  • Author(s): Yamazaki, Y.
  • Organizer: XXIst Congress of the International Society on Thrombosis and Haemostasis
  • Place of Presentation: Geneva, Switzerland
• [Presentation] Induction of vascular leakage and plasma clotting by a serine proteinase from Aeromonas sobria (2007)
  • Author(s): Imamura, T
  • Organizer: Xth International Symposium on Proteinase Inhibitors and Biological Control
  • Place of Presentation: Portoroz, Slovenia
• [Presentation] Induction of coagulation by a serine protease from Aeromonas sobria through activation of prothrombin (2007)
  • Author(s): Nitta, H
  • Organizer: 8th World Congress of Inflammation
  • Place of Presentation: Copenhagen, Denmark
• [Presentation] Aeromonas sobriaセリンプロテアーゼの凝固・線溶作用とその機序 (2007)
  • Author(s): 今村隆寿
  • Organizer: 第96回日本病理学会総会
  • Place of Presentation: 大阪
• [Presentation] Induction of vascular leakage and plasma clotting by a serine proteinase from Aeromonas sobria (2007)
  • Author(s): Imamura, T.
  • Organizer: Xth International Symposium on Proteinase Inhibitors and Biological Control
  • Place of Presentation: Portoroz,Slovenia
• [Presentation] Production of C5a by a serine proteinase from Aeromonas sobria,leading to Neutrophil Migration and vascular leakage (2007)
  • Author(s): Imamura, T.
  • Organizer: 5th International Symposium of Proteolysis
  • Place of Presentation: Patras,Greece
• [Presentation] 尿を用いた腎炎病態のリアルタイム解析-トロンビン活性の重要性- (2006)
  • Author(s): 北本康則
  • Organizer: 第53回日本臨床検査医学会学術集会
  • Place of Presentation: 弘前
  • Year and Date: 2006-11-11
• [Presentation] 腎炎治療のリアルタイムマーカー, 尿トロンビン (2006)
  • Author(s): 北本康則
  • Organizer: 第51回日本腎臓学会学術総会
  • Place of Presentation: 福岡
  • Year and Date: 2006-05-30
• [Presentation] 尿トロンビン解析におよぼす腎組織, 血管内, および尿中での血液凝固反応とその制御 (2006)
  • Author(s): 北本康則
  • Organizer: 第50回日本腎臓学会学術総会
  • Place of Presentation: 浜松
  • Year and Date: 2006-05-27
• [Presentation] Aeromonas sobriaセリンプロテアーゼによる血漿漏出とその機序 (2006)
  • Author(s): 今村隆寿
  • Organizer: 第11回病態と治療におけるプロテアーゼとインヒビター研究会
  • Place of Presentation: 仙台
• [Presentation] Induction of blood coagulation and fibrinolysis by cysteine proteinases from Staphylococcus aureus : A new mechanism of disseminated intravascular coagulation (2006)
  • Author(s): Imamura, T.
  • Organizer: 5th International Conference on Cysteine Proteinases and Their Inhibitors : From structure to regulation and biology
  • Place of Presentation: Portoroz, Slovenia
• [Presentation] Urokinase suppresses mononuclear and eosinophilic leukocyte infiltration in a rabbit model (2006)
  • Author(s): Lai. Z.-F
  • Organizer: 46th Annual Meeting of The American Society for Cell Biology
  • Place of Presentation: San Diego, USA
• [Presentation] Aeromonas sobriaセリンプロテアーゼのキニン遊離を介した血管漏出と血圧低下作用 (2006)
  • Author(s): 今村隆寿
  • Organizer: 第80回日本細菌学会総会
  • Place of Presentation: 大阪
• [Presentation] Snake venom VEGF-F is a most potent inducer of vascular permeability (2006)
  • Author(s): Yamazaki, Y
  • Organizer: XVth World Congress on Animal, Plant and Microbial Toxins
  • Place of Presentation: Glasgow, UK
• [Presentation] Aeromonas sobriaセリンプロテアーゼの血管透過性亢進作用とその機序 (2006)
  • Author(s): 今村隆寿
  • Organizer: 第95回日本病理学会総会
  • Place of Presentation: 東京
• [Presentation] 尿トロンビン、CD14+16+細胞、oil red陽性細胞測定による増殖性腎炎のネフロン傷害の解析 (2005)
  • Author(s): 北本康則
  • Organizer: 第49回日本腎臓学会学術総会
  • Place of Presentation: 東京
• [Book] 炎症・再生医学事典 カリクレイン・キニン系 (2009)
  • Author(s): 今村隆寿
  • Publisher: 朝倉書店出版
• [Book] カリクレイン・キニン系. 炎症・再生医学 (2008)
  • Author(s): 今村隆寿
  • Publisher: 朝倉書店出版
• [Remarks]
  • URL: http://www.kumamoto-u.ac.jp/seeds/seeds/61580016/index.html
• [Remarks]
  • URL: http://www.kumamoto-u.ac.jp/seeds/seeds/61580016/index.html
• [Patent(Industrial Property Rights)] 癌細胞C5a産生プロテアーゼおよびC5a受容体をターゲットにした癌治療法 (2009)
  • Inventor(s): 今村隆寿、新田英利
  • Industrial Property Rights Holder: 熊本大学長
  • Filing Date: 2009-03-06