天然由来シグナル伝達阻害物質の探索と医薬への応用

• Project/Area Number: 16073217
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Science and Engineering
• Research Institution: Keio University
• Principal Investigator: 梅澤 一夫 (梅沢 一夫) Keio University, 理工学部, 教授 (70114402)
• Co-Investigator(Kenkyū-buntansha): 鈴木 芳和 慶應義塾大学, 理工学部, 助手 (00348822)
• Project Period (FY): 2004 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥28,300,000 (Direct Cost: ¥28,300,000); Fiscal Year 2007: ¥8,100,000 (Direct Cost: ¥8,100,000); Fiscal Year 2006: ¥8,100,000 (Direct Cost: ¥8,100,000); Fiscal Year 2005: ¥8,100,000 (Direct Cost: ¥8,100,000); Fiscal Year 2004: ¥4,000,000 (Direct Cost: ¥4,000,000)
• Keywords: 癌 / 生理活性物質 / シグナル伝達 / 微生物 / 有機化学 / 生理活性 / NF-κB / エポキシキノマイシン / DHMEQ / コノフィリン / インスリン / ヘプタデプシン / リポ多糖

Research Abstract

今年度はNF-κB阻害物質(-)-DHMEQの標的分子を解明することができた。1)(-)-DHMEQの標的分子の解明。私達が発見した(-)-DHMEQは様々な病態動物モデルで強い治療効果を示した。しかし、その標的分子は未解明であった。今回、(-)-DHMEQはp65に直接結合して、DNA結合能を失わせることを明らかにした。
recombinant p65に対して(-)-DHMEQは1:1量で、in vitroでDNA結合を抑制した。SPRにおいてresponseがあり、MALDI-TOF MSにおいて1:1の共有結合が示された。さらに特定のアミノ酸に結合することもわかった。さらに制限プロテアーゼを用いたprotein mass fingerprintを応用したMALDI-TOF MS測定からも特定のアミノ酸への結合が示された。以上より、(-)-DHMEQはNF-κB構成因子の特定のアミノ酸に共有結合することでNF-κBのDNA結合能を抑制することがわかった。この結果は、より有効なNF-κB阻害剤創製の指標となると考えられる(論文投稿中)。2) conophyllineの標的分子ARL61Pの機能を調べるためにSiRNAによるノックダウンマウスを作成した。3) 新しいNF-κB阻害剤の発見。微生物よりNO産生阻害活性、NF-κB阻害活性のある既知物質を微生物から得た(論文準備中)。4) (-)-DHMEQを光学分割する方法として酵素リパーゼを用いる方法を見出したが、その溶液をかえることでより効率よく光学活性体が得られるようになった。5) (-)-DHMEQによる癌細胞の転移・浸潤能の抑制。高転移性乳癌細胞であるMDA-MB-231細胞において(-)-DHMEQは、CXCR4の発現を抑制することにより、転移部位へのchemotaxisを抑制するほか、autocrine機構により自身の遊走・浸潤能を低下させることが示唆された。

Research Products

• [Journal Article] NF-κBを標的とする抗癌剤 (2008)
  • Author(s): 鈴木絵里子、梅澤一夫
  • Journal Title: 医学のあゆみ 244 Pages: 65-69
• [Journal Article] Conophylline and betacellulin-δ4 : an effective combination of differentiation factors for pancreatic β cells. (2007)
  • Author(s): R. Kitamura, K. Umezawa, et al.
  • Journal Title: Endocline J 54 Pages: 255-264
  • NAID: 10019478673
  • Peer Reviewed
• [Journal Article] IκBα independent induction of NF-κB and its inhibition by DHMEQ in Hodgkin/Reed-Sternberg cells. (2007)
  • Author(s): M. Watanabe, K. Umezawa, et al.
  • Journal Title: Laboratory Investigation 87 Pages: 372-382
  • Peer Reviewed
• [Journal Article] Reversal of resistance to cytotoxic cancer therapies : DHMEQ as a chemo-sensitizing and immuno-sensitizing agent. (2007)
  • Author(s): A. Katsman, K. Umezawa, et al.
  • Journal Title: Drug Resistance Updates 10 Pages: 1-12
• [Journal Article] Suppression of diabetes-induced retinal inflammation by blocking angiotensin II type 1 receptor or its downstream NF-κB pathway. (2007)
  • Author(s): N. Nagai, K. Umezawa, et al.
  • Journal Title: Investigative Ohthalmology and Visual Science 48 Pages: 4342-4350
  • Peer Reviewed
• [Journal Article] Targeting NF-κB and induction of apoptosis by novel NF-κB inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ) in Burkitt lymphoma cells. (2007)
  • Author(s): N. Kimura, K. Umezawa, et al.
  • Journal Title: Leukemia Research 31 Pages: 6184-6189
  • Peer Reviewed
• [Journal Article] Rituximab inhibits the constitutively activated PI3K-Akt pathway in B-NHL cell lines : Involvement in chemosensitization to drug-induced apoptosis (2007)
  • Author(s): E. Suzuki, K. Umezawa, et al.
  • Journal Title: Oncogene 26 Pages: 6184-6189
  • Peer Reviewed
• [Journal Article] Tyrosine kinase inhibitor methyl-2,5-dihydroxycinnamate induces PML nuclear body formation and apoptosis in tumor cells. (2007)
  • Author(s): N. Komura K. Umezawa, et al.
  • Journal Title: Exp. Cell Res. 313 Pages: 2753-2765
  • Peer Reviewed
• [Journal Article] Dehydroxymethylepoxyquinomicin (DHMEQ) therapy reduces tumor formation in mice inoculated with Tax-deficient adult T-cell leukemia-derived cell lines. (2007)
  • Author(s): T. Ohsugi, K. Umezawa, et al.
  • Journal Title: Cancer Letters 257 Pages: 206-215
  • Peer Reviewed
• [Journal Article] Inflammatory stimuli from macrophages and cancer cells synergistically promote tumor growth and angiogenesis. (2007)
  • Author(s): Y. N. Kimura, K. Umezawa, et al.
  • Journal Title: Cancer Science 98 Pages: 2009-2018
  • NAID: 10019996850
  • Peer Reviewed
• [Journal Article] Inhibition of tumor angiogenesis by targeting endotheliai ATP synthase with sangivamycin. (2007)
  • Author(s): Y. Komi, K. Umezawa, et al.
  • Journal Title: Japan Journal of Clinical Oncology 37 Pages: 867-873
  • Peer Reviewed
• [Journal Article] NF-κB inhibitorDHMEQ suppresses osteoclastogenesis and expression of NFATc1 in mouse arthritis without affecting expression of RANCL, OPG or M-CSF. (2007)
  • Author(s): T. Kubota, K. Umezawa, et al.
  • Journal Title: Arthritis Research & Therapy 9/5/R97(Open Access)
  • Peer Reviewed
• [Journal Article] 炎症を抑制する生理活性物質の探索 (2007)
  • Author(s): 梅澤一夫
  • Journal Title: 蛋白質 核酸 酵素 52, No.13 Pages: 1685-1689
• [Journal Article] Designed ATRA analogue active against ATRA-resistant acute promyelocytic leukemia cells having a single nucleotide substitution in their retinoic acid receptor. (2007)
  • Author(s): N. Komura, K. Umezawa, et al.
  • Journal Title: Leuk. Res. 31 Pages: 301-313
  • Peer Reviewed
• [Journal Article] Designed ATRA analogue active against ATRA-resistant acute promyelocytic leukemia cells having a single nucleotide substitution in their retinoic acid receptor. (2007)
  • Author(s): N.Komura, K.Umezawa, et al.
  • Journal Title: Leuk. Res. 31 Pages: 301-313
• [Journal Article] In vitro and in vivo antitumor activity of the NF-kappa B inhibitor DHMEQ in the human T-cell leukemia virus type I-infected cell line, HUT-102. (2006)
  • Author(s): T.Ohsugi, K.Umezawa, et al.
  • Journal Title: Leuk. Res. 30 Pages: 90-97
• [Journal Article] Inhibition of Tipopolysaccharide activity by a bacterial cyclic lipopeptide surfactin. (2006)
  • Author(s): T.Takahashi, K.Umezawa, et al.
  • Journal Title: J. Antibiot 59 Pages: 35-43
• [Journal Article] DHMEQ, a new NF-κB inhibitor, induces apoptosis and enhances fludarabine effects on chronic lymphocytic leukemia cells. (2006)
  • Author(s): R.Horie, K.Umezawa, et al.
  • Journal Title: Leukemia 20 Pages: 800-806
• [Journal Article] Inhibition of macrophage activation and phagocytosis by a novel NF-κB inhibitor, dehydroxymethylepoxyquinomicin. (2006)
  • Author(s): E.Suzuki, K.Umezawa
  • Journal Title: Biomed. Pharmacother. 60 Pages: 578-586
• [Journal Article] Inhibition of NF kappa B activation by 9-methylstreptimidone isolated from Streptomyces. (2006)
  • Author(s): Z.Wang, K.Umezawa, et al.
  • Journal Title: Heterocycles 69 Pages: 377-383
• [Journal Article] 転写因子NF-κBと癌の進展。 (2006)
  • Author(s): 梅澤一夫
  • Journal Title: バイオサイエンスとインダストリー 64 (No.4) Pages: 224-225
• [Journal Article] Conophylline : A novel differentiation inducer for pancreatic β cells. (2006)
  • Author(s): I.Kojima, K.Umezawa
  • Journal Title: Int. J. Biochem. 38 Pages: 923-930
• [Journal Article] Survivin associates with cell proliferation in renal cancer cells : Regulation of surviving expression by insulin-like growth factor-1, interferon-γ and a novel NF-κB inhibitor. (2006)
  • Author(s): A.Sato, K.Umezawa, et al.
  • Journal Title: Int. J. Oncology 28 Pages: 841-846
• [Journal Article] Blocking NF-κB as a potential strategy to treat adult T-cell leukemia/lymphoma. (2006)
  • Author(s): R.Horie, K.Umezawa, et al.
  • Journal Title: Drug News Persrect. 19 Pages: 201-209
• [Journal Article] ケミカルパソロシーによる病態解析と薬剤開発。 (2006)
  • Author(s): 梅澤一夫
  • Journal Title: 現代化学2006年7月号 No.424 Pages: 38-42
• [Journal Article] Adrenaline contributes to prenatal respiratory maturation in rat medulla-spinal cord preparation. (2006)
  • Author(s): M.Fujii, K.Umezawa, et al.
  • Journal Title: Brain Res. 1090 Pages: 45-50
• [Journal Article] Inhibition of tumor growth by NF-κB inhibitors. (2006)
  • Author(s): K.Umezawa
  • Journal Title: Cancer Science 97 Pages: 990-995
  • NAID: 10019377732
• [Journal Article] Effects of a novel NF-κB inhibitor, dehydroxymethyepoxyquinomicin (DHMEQ), on growth, apoptosis gene expression, and chemosensitivity in head and neck squamous cell carcinoma cell lines. (2006)
  • Author(s): H.Ruan, K.Umezawa, et al.
  • Journal Title: Leukemia 20 Pages: 800-806
• [Journal Article] BRAFV600E promotes invasiveness of thyroid cancer cells through NF-κB activation. (2006)
  • Author(s): I.Palona, K.Umezawa, et al.
  • Journal Title: Endocrinology 147 Pages: 5699-5707
• [Journal Article] Control of allograft rejection by applying a novel NF-κB inhibitor, dehydroxymethylepoxyquinonicin. (2006)
  • Author(s): S.Ueki, K.Umezawa, et al.
  • Journal Title: Transplantation 82 Pages: 1720-1727
• [Journal Article] Nitric oxide donors induce neurotrophin-like survival signaling and protect neurons against apoptosis. (2005)
  • Author(s): C.Culmsee, K.Umezawa, et al.
  • Journal Title: Molecular Pharmacology 68 Pages: 1006-1017
• [Journal Article] Dopamine desynchronizes the pace-making neuronal zctivity of rat respiratory rhythm generation. (2005)
  • Author(s): M.Fujii, K.Umezawa, et al.
  • Journal Title: Eur. J. Neurosci. 23 Pages: 1015-1027
• [Journal Article] Reversal of drug resistance by COTC based on intracellular glutathione and glyoxalase I. (2005)
  • Author(s): D.Kamiya et al.
  • Journal Title: Biorg.Med.Chem.Lett 15 Pages: 1111-1114
• [Journal Article] Inhibition of RANKL-induced osteoclastogenesis by (-)-DHMEQ, a novel NF-κB inhibitor, through downregulation of NFATc1. (2005)
  • Author(s): H.Takatsuna et al.
  • Journal Title: J.Bone Mineral Res 20 Pages: 653-661
• [Journal Article] Inhibition of inflammatory cytokine secretion from mouse microglia cells by DHMEQ, an NF-κB inhibitor. (2005)
  • Author(s): H.Takatsuna et al.
  • Journal Title: Biomed.Pharmacother 59 Pages: 318-322
• [Journal Article] Inhibition of cellular adhesion in human umbilical vein endothelial cells by NF-κB inhibitor DHMEQ under flow. (2005)
  • Author(s): O.Ohno et al.
  • Journal Title: Oncology Res. 15 Pages: 189-198
• [Journal Article] A novel nuclear factor kappa B inhibitor induced apoptosis of multiple myeloma cells. (2005)
  • Author(s): H.Tatetsu et al.
  • Journal Title: Mol.Cancer Ther 4 Pages: 1114-1120
• [Journal Article] Effect of a small molecule inhibitor of nuclear factor-κB nuclear translocation in a murine model of arthritis and cultured human synovial cells. (2005)
  • Author(s): K.Wakamatsu et al.
  • Journal Title: Arthritis Research & Therapy 7 Pages: 1348-1359
• [Journal Article] Dual targeting of transformed and untransformed HTLV-1-infected T cells by DHMEQ, a potent and selective inhibitor of NF-κB, as a strategy for chemoprevention and therapy of adult T cell leukemia. (2005)
  • Author(s): M.Watanabe et al.
  • Journal Title: Blood 106 Pages: 2462-2471
• [Journal Article] I JunB induced by constitutive CD30-extracellular signal-regulated kinase /2 mitogen-activated protein kinase signaling activates the CD30 promoter in anaplastic large cell lymphoma and Reed-Sternberg cells of Hodgkin lymphoma. (2005)
  • Author(s): M.Watanabe et al.
  • Journal Title: Cancer Res 65 Pages: 7628-7634
• [Journal Article] Prevention of cancer cachexia by a novel nuclear factor κB inhibitor in prostate cancer. (2005)
  • Author(s): K.Kuroda et al.
  • Journal Title: Clin.Cancer Res 11 Pages: 5590-5594
• [Journal Article] In vivo antitumor activity of the NF-κB inhibitor dehydroxymethyl-epoxyquinomicin in a mouse model of adult T-cell leukemia. (2005)
  • Author(s): T.Ohsugi et al.
  • Journal Title: Carcinogenesis 26 Pages: 1382-1388
• [Journal Article] Nitric oxide donors induce neurotrophin-like survival signaling and protect neurons against apoptosis. (2005)
  • Author(s): C.Culmsee et al.
  • Journal Title: Molecular Pharmacology 68 Pages: 1006-1017
• [Journal Article] Enhancement of the caspase-independent apoptotic sensitivity of pancreatic cancer cells by DHMEQ, an NF-κB inhibitor. (2005)
  • Author(s): G.Matsumoto et al.
  • Journal Title: Int.J.Oncology 27 Pages: 1247-1255
• [Journal Article] Dopaminergic effects on respiratory rhythm generation in rat brainstem-spinal cord preparation (2005)
  • Author(s): M.Fujii, K.Umezawa, A.Arata
  • Journal Title: Neuroscience Res. 50 Pages: 355-359
• [Journal Article] Targeting of NF-κB pathways by DHMEQ, a novel inhibitor of breast carcinomas : Anti-tumor and anti-angiogenic activity in vivo (2005)
  • Author(s): G.Matsumoto, J.Namekawa, M.Muta, T.Nakamura, H.Bando, K.Tohyama, M.Toi, K.Umezawa
  • Journal Title: Clin.Cancer Res. 11 Pages: 1287-1293
• [Journal Article] Synergistic antiviral effect of oxanosine and ddI against human immunodeficiency virus (2005)
  • Author(s): M.Nakamura, T.Ogawa, A.Yokono, S.Nakamori, T.Ohno, K.Umezawa
  • Journal Title: Biomed.Pharmacother. 59 Pages: 47-50
• [Journal Article] A novel NF-κB inhibitor DHMEQ selectively targets constitutive NF-κB activity and induces apoptosis of multiple myeloma cells in vitro and in vivo (2005)
  • Author(s): M.Watanabe, M.Z.Dewan, T.Okamura, M.Sasaki, K.Itoh, M.Higashihara, H.Mizoguchi, M.Honda, T.Sata, T.Watanabe, N.Yamamoto, K.Umezawa, R.Horie
  • Journal Title: Int.J.Cancer 114 Pages: 32-38
• [Journal Article] Inhibition of RANKL-induced osteoclastogenesis by (-)-DHMEQ, a novel NF-κB inhibitor, through downregulation of NFATc1 (2005)
  • Author(s): H.Takatsuna, M.Asagiri, T.Kubota, K.Oka, T.Osada, C.Sugiyama, H.Saito, K.Aoki, K.Ohya, H.Takayanagi, K.Umezawa
  • Journal Title: J.Bone Mineral Res. 20 Pages: 653-661
• [Journal Article] Genistein and daizein induce cell proliferation and their metabolites cause oxidative DNA damage in relation to isoflavone-induced cancer of estrogen-sensitive organs (2004)
  • Author(s): M.Murata, K.Midorikawa, M.Koh, K.Umezawa, S.Kawanishi
  • Journal Title: Biochemistry 43 Pages: 2569-2577
• [Journal Article] Preparation and biological activities of optically active dehydroxymethylepoxyquinomicin, a novel NF-κB inhibitor. (2004)
  • Author(s): Y.Suzuki, C.Sugiyama, O.Ohno, K.Umezwa
  • Journal Title: Tetrahedron 60 Pages: 7061-7066
• [Journal Article] Isolation of heptadepsin, a novel bacterial cyclic depsipeptide that inhibits lipopolysaccharide activity (2004)
  • Author(s): O.Ohno, Y.Ikeda, R.Sawa, M.Igarashi, N.Kinoshita, Y.Suzuki, K.Miyake, K.Umezawa
  • Journal Title: Chemistry & Biology 11 Pages: 1059-1070
• [Journal Article] Promotion of β cell differentiation by conophylline in fetal and neonatal rat pancreas (2004)
  • Author(s): T.Ogata, L.Li, S.Yamada, Y.Yamamoto, Y.Tanaka, I.Takei, K.Umezawa, I.Kojima
  • Journal Title: Diabetes 53 Pages: 2596-2602
• [Journal Article] Screening of bioactive metabolites for pancreatic regeneration chemotherapy (2004)
  • Author(s): H.Takatsuna, K.Umezawa
  • Journal Title: Biomed.Pharmacother. 58 Pages: 610-613
• [Journal Article] Diabetes mellitus : rational basis, clinical approach and future therapy (2004)
  • Author(s): H.Ohgawara, K.Edamura, M.Kawakami, K.Umezawa
  • Journal Title: Biomed.Pharmacother. 58 Pages: 605-609
• [Journal Article] Induction of thyroid cancer cell apoptosis by a novel nuclear factor kappaB inhibitor, DHMEQ (2004)
  • Author(s): D.V.Starenki, H.Namba, V.A.Saenko, A.Ohtsuru, S.Maeda, K.Umezawa, S.Yamashita
  • Journal Title: Clin.Cancer Res. 10 Pages: 6821-6829
• [Journal Article] Inhibition of inflammatory cytokine secretion from mouse microglia cells by DHMEQ, an NF-κB inhibitor
  • Author(s): H.Takatsuna, S.Morita, T.Nagatsu, M.Sawada, K.Umezawa
  • Journal Title: Biomed.Pharmacother. (印刷中)
• [Journal Article] Reversal of drug resistance by COTC based on intracellular glutathione and glyoxalase
  • Author(s): D.Kamiya, Y.Uchihata, E.Ichikawa, K.Kato, K.Umezawa
  • Journal Title: Biorg.Med.Chem.Lett. (印刷中)
• [Journal Article] Inhibition of cellular adhesion in human umbilical vein endothelial cells by NF-κB inhibitor DHMEQ under flow
  • Author(s): O.Ohno, Y.Shima, Y.Ikeda, Y.Kawai, K.Sakurai, K.Watanabe, K.Umezawa
  • Journal Title: Oncology Res. (印刷中)
• [Presentation] NF-κB as a target of anticancer agents : Anticancer activity and molecular target of an NF-κB inhibitor, DHMEQ.(シンポジウム講演) (2007)
  • Author(s): K. Umezawa
  • Organizer: 第66回日本癌学会学術総会
  • Place of Presentation: 横浜
• [Book] 腐敗と再生 (2004)
  • Author(s): 梅澤一夫
  • Total Pages: 15
  • Publisher: 慶應義塾大学出版会
• [Remarks]
  • URL: http://www.applc.keio.ac.jp/~umezawa
• [Patent(Industrial Property Rights)] 抗腫瘍剤に対する腫瘍細胞の感受性増強剤 (2006)
  • Inventor(s): 梅澤一夫他3名
  • Industrial Property Rights Holder: 慶應義塾
  • Industrial Property Number: 2006-274257
  • Filing Date: 2006-10-05
• [Patent(Industrial Property Rights)] NF-κBを阻害する抗炎症、抗癌剤 (2006)
  • Inventor(s): 梅澤一夫ほか
  • Industrial Property Rights Holder: 慶應義塾
  • Industrial Property Number: 2006-000902
  • Filing Date: 2006-01-05