酸素センサー分子Keap-1の活性阻害薬の探索:酸化ストレスの薬物制御

• Project/Area Number: 21659213
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Kidney internal medicine
• Research Institution: Tohoku University
• Principal Investigator: 宮田 敏男 Tohoku University, 大学院・医学系研究科, 教授 (10222332)
• Project Period (FY): 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥3,100,000 (Direct Cost: ¥3,100,000); Fiscal Year 2009: ¥3,100,000 (Direct Cost: ¥3,100,000)
• Keywords: 酸化ストレス応答 / Nrf2-Keep1相互作用 / keep1阻害剤 / X線結晶解析情報 / レポーターアッセイ / SBDD / ジメチルマレイン酸 / edaravone

Research Abstract

その下流に多くの抗酸化ストレス遺伝子を擁するNrf2は異物代謝・酸化ストレス応答の鍵となる転写活性化因子であり、Keap1によって負の制御を受けている。本研究はKeap1に結合し、Nrf2-Keap1相互作用を阻害することでNrf2の分解を抑制し、Nrf2並びに抗酸化ストレス遺伝子の活性化をもたらす新規低分子化合物をstructure based drug design (SBDD)に基づくvirtual screeningとvitro生物学的評価法により探索して、病態解明のみならず治療コンセプトの検証に用いるための候補化合物の選定を目的とした。
Keap1分子のX線結晶解析情報を基に、その活性調節部位に挿入し活性を阻害する低分子ヒット化合物をSBDDで探索し、該当する化合物から購入あるいは合成できた37化合物を検体とした。化合物のvitro生物学的評価は、Nrf2 locusにLacZをノックインしてNeh2-LacZ融合タンパク質が発現するようになったマウス繊維芽細胞を使用してレポーターアッセイを行い、DEM(ジメチルマレイン酸)を陽性対照として、検体によるβ-galacosidase活性をscreeningした。
結果は、37化合物中、陽性対照のDEMとほぼ匹敵する活性を示す1化合物(TM-8009)が得られ、期初の目標は達成された。また、抗酸化作用等が知られる既存薬や臨床候補化合物のscreeningも行い、edaravoneにも同様の活性があることが確認された。
今後、それらの新たに見出された化合物をリードとして構造最適化を図り、vivoでの治療コンセプトの検証をしたいと考えている。

Research Products

• [Journal Article] Prevention and treatment of diabetic nephropathy : new approaches (2010)
  • Author(s): Miyata T, van Ypersele de Strihou C.
  • Journal Title: Review of Endocrinology (未定, In press)
  • Peer Reviewed
• [Journal Article] A novel inhibitor of plasminogen activator inhibitor-1 provides anti-thrombotic benefits devoid of bleeding effect in non-human primates (2010)
  • Author(s): Izuhara Y, Yamaoka N, Kodama H, Dan T, Takizawa S, Hirayama N, Meguro K, van Yperselede Strihou Charles, Miyata T.
  • Journal Title: J Cereb Blood Flow Metab 30 Pages: 904-912
  • Peer Reviewed
• [Journal Article] Enhanced carbonyl stress in a subpopulation of schizophrenia (2010)
  • Author(s): Arai M, Yuzawa H, Nohara I, Ohnishi T, Obata N, Iwayama Y, Haga S, Toyota T, Ujike H, Arai M, Ichikawa T, Nishida A, Tanaka Y, Furukawa A, Aikawa Y, Kuroda O, Niizato K, Izawa R, Nakamura K, Mori N, Matsuzawa D, Hashimoto K, Iyo M, Sora I, Matsushita M, Okazaki Y, Yoshikawa T, Miyata T, Itokawa M.
  • Journal Title: Arch Gen Psychiatry (未定, In press)
  • Peer Reviewed
• [Journal Article] Effect of edaravone on the estimated glomerular filtration rate (eGFR) in acute ischemic stroke patients with chronic kidney disease (2010)
  • Author(s): Tsukamoto Y, Takizawa S, Takahashi W, Mase H, Miyachi H, Miyata T, Takagi S.
  • Journal Title: J Stroke Cerebrovasc Dis (未定, In press)
  • Peer Reviewed
• [Journal Article] Relationship of Skin-autofluorescence to Cardiovascular Disease in Japanese Hemodialysis Patients (2010)
  • Author(s): Tanaka K, Katoh T, Asai J, Nemoto F, Suzuki H, Asahi K, Sato K, Sakaue M, Miyata T, Watanabe T.
  • Journal Title: Ther Apher Dial (未定, In press)
  • Peer Reviewed
• [Journal Article] Body weight control by a high-carbohydrate/low-fat diet slows the progression of diabetic kidney damage in an obese, hypertensive, type 2 diabetic rat model (2010)
  • Author(s): Ohtomo S, Izuhara Y, Nangaku M, Dan T, Ito S, Van Ypersele de Strihou C, Miyata T.
  • Journal Title: J Obesity (未定, In press)
  • Peer Reviewed
• [Journal Article] Hyperglycaemia-induced impairment of endothelium-dependent vasorelaxation in rat mesenteric arteries is mediated by intracellular methylglyoxal levels in a pathway dependent on oxidative stress (2010)
  • Author(s): Brouwers O, Niessen PM, Haenen G, Miyata T, Brownlee M, Stehouwer CD, De Mey JG, Schalkwijk CG.
  • Journal Title: Diabetologia 53 Pages: 989-1000
  • Peer Reviewed
• [Journal Article] Advanced glycation end-products, anti hypertensive treatment and diastolic function in patients with hypertension and diastolic dysfunction (2010)
  • Author(s): Hartog JW, van de Wal RM, Schalkwijk CG, Miyata T, Jaarsma W, Plokker HW, van Wijk LM, Smit AJ, van Veldhuisen DJ, Voors AA.
  • Journal Title: Eur.J.Heart Failure 12 Pages: 397-403
  • Peer Reviewed
• [Journal Article] Metallothionein is upregulated by hypoxia and stabilizes hypoxia-inducible factor in the kidney (2009)
  • Author(s): Kojima I, Tanaka T, Inagi R, Nishi H, Aburatani H, Kato H, Miyata T, Fujita T, Nangaku M.
  • Journal Title: Kidney Int 75 Pages: 268-277
  • Peer Reviewed
• [Journal Article] Advanced glycation end products in patients with cerebral infarction (2009)
  • Author(s): Ohnuki Y, Nagano R, Takizawa S, Takagi S, Miyata T.
  • Journal Title: Intern Med 48 Pages: 587-591
  • NAID: 130000108031
  • Peer Reviewed
• [Journal Article] Translation of basic science into clinical medicine : Novel targetsfor diabetic nephropathy (2009)
  • Author(s): Miyata T, van Ypersele de Strihou C.
  • Journal Title: Nephrol Dial Transplant 24 Pages: 1373-1377
  • Peer Reviewed
• [Journal Article] Glyoxalase I overexpression ameliorates renal ischemia-reperfusion injury in rats (2009)
  • Author(s): Kumagai T, Nangaku M, Kojima I, Nagai R, Julie R.Ingelfinger, Miyata T, Fujita T, Inagi R.
  • Journal Title: Am J Physiol-Renal Physiol 296
  • Peer Reviewed
• [Journal Article] Translational medicine : Novel mechanisms and therapeutic options in diabetic nephropathy (2009)
  • Author(s): Miyata T.
  • Journal Title: Pol Arch Med Wewn 119 Pages: 261-264
  • Peer Reviewed
• [Journal Article] Effects of NXY-059 in experimental stroke : an individual animal meta-analysis (2009)
  • Author(s): Bath PM, Gray LJ, Bath AJ, Buchan A, Miyata T, Green AR : NXY-059 Efficacy Meta-analysis in Individual Animals with Stroke Investigators
  • Journal Title: British J Pharmacology 157 Pages: 1157-1171
  • Peer Reviewed
• [Journal Article] Very high doses of valsartan provide renoprotection independently of blood pressure in a type 2 diabetic nephropathy rat model (2009)
  • Author(s): Tominaga N, Robert A, Izuhara Y, Ohtomo S, Dan T, Chihara K, Kurokawa K, van Ypersele de Strihou C Miyata T.
  • Journal Title: Nephrology 14 Pages: 581-587
  • Peer Reviewed
• [Journal Article] A sartan derivative with a very low angiotensin II receptor affinity ameliorates ischemic cerebral damage (2009)
  • Author(s): Takizawa S, Uesugi T, Nagata E, Takagi S, van Ypersele de Strihou C, Miyata T.
  • Journal Title: J Cereb Blood Flow Metab 29 Pages: 1665-1672
  • Peer Reviewed
• [Journal Article] Angiotensin II type 1A receptor deficiency and longevity (2009)
  • Author(s): Nishiyama A, Matsusaka T, Miyata T.
  • Journal Title: Nephrol Dial Transplant 24 Pages: 3280-3281
  • Peer Reviewed
• [Journal Article] Methylglyoxal contributes to the development of insulin resistance and salt sensitivity in Sprague Dawley rats (2009)
  • Author(s): Qi G, Mori T, Jiang Y, Hu C, Osaki Y, Yoneki Y, Sun Y, Hosoya T, Kawamata A, Ogawa S, Nakayama M, Miyata T, Ito S.
  • Journal Title: J Hyperten 27 Pages: 1664-1671
  • Peer Reviewed
• [Journal Article] Inhibition of prolyl hydroxylase domain-containing protein suppressed 1 ipopolysaccharide-induced TNF-a expression (2009)
  • Author(s): Takeda K, Ichiki T, Narabayashi E, Inanaga K, Miyazaki R, Hashimoto T, Matsuura H, Ikeda J, Miyata T, Sunagawa K.
  • Journal Title: Arterio Thromb Vas Biol 29 Pages: 2132-2137
  • Peer Reviewed
• [Journal Article] Diabetic nephropathy : a disorder of oxygen metabolism? (2009)
  • Author(s): Miyata T, van Ypersele de Strihou C.
  • Journal Title: Nat Rev Nephrol 6 Pages: 83-95
  • Peer Reviewed
• [Presentation] 糖尿病性腎症と酸素代謝異常 (2010)
  • Author(s): 宮田敏男
  • Organizer: レドックス生命科学第170委員会第22回研究会
  • Place of Presentation: 大阪
  • Year and Date: 2010-03-19
• [Presentation] RAS阻害薬の臓器保護について (2010)
  • Author(s): 宮田敏男
  • Organizer: 苫小牧ラジレス講演会
  • Place of Presentation: 苫小牧
  • Year and Date: 2010-03-11
• [Presentation] RAS阻害薬の臓器保護について (2010)
  • Author(s): 宮田敏男
  • Organizer: 白石医師会講演会
  • Place of Presentation: 仙台
  • Year and Date: 2010-03-09
• [Presentation] 大学からの創薬 (2010)
  • Author(s): 宮田敏男
  • Organizer: 東北大学国際産学連携シンポジウム
  • Place of Presentation: 東京
  • Year and Date: 2010-02-22
• [Presentation] 糖尿病性腎症と酸素代謝異常 (2010)
  • Author(s): 宮田敏男
  • Organizer: 日本内科学会東北支部・第55回生涯教育講演会
  • Place of Presentation: 仙台
  • Year and Date: 2010-02-20
• [Presentation] DRIのポテンシャル-降圧と独立した腎臓保護- (2010)
  • Author(s): 宮田敏男
  • Organizer: DRI FORUM
  • Place of Presentation: 横浜
  • Year and Date: 2010-02-03
• [Presentation] Development of novel therapeutic agents for the treatment of renal car diovascular diseases : inhibitors of plasminogen inhibitor 1 and oxygen sensor prolylhydroxylase (2009)
  • Author(s): 宮田敏男
  • Organizer: The 1st International Symposium～Challenge to Medical Innovation～
  • Place of Presentation: 仙台
  • Year and Date: 2009-12-07
• [Presentation] RAS阻害薬の臓器保護について (2009)
  • Author(s): 宮田敏男
  • Organizer: Rasilez Expert Meeting in Sapporo
  • Place of Presentation: 札幌
  • Year and Date: 2009-11-28
• [Presentation] 増加する腎臓病-如何に早期に診断し治療するか (2009)
  • Author(s): 宮田敏男
  • Organizer: 町田RAAフォーラム
  • Place of Presentation: 東京
  • Year and Date: 2009-11-19
• [Presentation] 糖尿病性腎症と酸素代謝異常 (2009)
  • Author(s): 宮田敏男
  • Organizer: 大阪CKDエキスパート会議
  • Place of Presentation: 大阪
  • Year and Date: 2009-11-13
• [Presentation] 増加する腎臓病-如何に早期に診断し治療するか (2009)
  • Author(s): 宮田敏男
  • Organizer: 福島県白河地区学術講演会
  • Place of Presentation: 福島
  • Year and Date: 2009-10-14
• [Presentation] 糖尿病性腎症と酸素代謝異常について (2009)
  • Author(s): 宮田敏男
  • Organizer: 第8回秋田県分子病態研究会
  • Place of Presentation: 秋田
  • Year and Date: 2009-10-09
• [Presentation] Abnormalities of oxygen metabolism in diabetic nephropathy ; Hypoxia and oxidative stress (2009)
  • Author(s): 宮田敏男
  • Organizer: 第21回腎とフリーラジカル研究会
  • Place of Presentation: 岡山
  • Year and Date: 2009-09-26
• [Presentation] Advanced glycation, hypoxia, and oxidative stress (2009)
  • Author(s): 宮田敏男
  • Organizer: World Congress of Nephrology
  • Place of Presentation: Milan, Italy
• [Presentation] Interplays among hypoxia, oxidative stress, and advanced glycation? (2009)
  • Author(s): 宮田敏男
  • Organizer: 10th International Symposium of the Maillard Reaction
  • Place of Presentation: Palm Cove, Australia
• [Presentation] COUP-TFIIは、Wnt/β-cateninシグナルの下流でPPARγの発現をエピジェネティックに調節することで脂肪細胞分化を制御する (2009)
  • Author(s): 岡村将史, 若林賢一, 堤修一, 宮田敏男, 内藤眞, 浜窪隆雄, 伊藤貞嘉, 油谷浩幸, 児玉龍彦, 酒井寿郎
  • Organizer: 第13回日本心血管内分泌代謝学会
  • Place of Presentation: 埼玉
• [Presentation] 大学からの創薬の一例-出血性副作用のない経口抗血栓薬(PAI-1阻害薬) (2009)
  • Author(s): 宮田敏男
  • Organizer: 第16回理事長ファンドワークショップ
  • Place of Presentation: 岐阜
• [Book] Oxidative Stress in Applied Basic Research and Clinical Practice : 'Renal Disorders' (2010)
  • Author(s): 宮田敏男, van Ypersele de Strihou C.
  • Publisher: Springer(印刷中)