がん増殖環境修飾物質の研究

• Project/Area Number: 12217157
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The Institute of Physical and Chemical Research (2003-2004); Microbial Chemistry Research Foundation (2000-2002)
• Principal Investigator: 長田 裕之 (2003-2004) 独立行政法人理化学研究所, 長田抗生物質研究室, 主任研究員 (80160836); 石塚 雅章 (2000-2002) 財団法人微生物化学研究会, 化学療法研究所, 所長 (80159722)
• Co-Investigator(Kenkyū-buntansha): 川田 学 財団法人微生物化学研究会, 微生物化学研究センター・沼津創薬医科学研究所・創薬基盤研究ユニット, ユニット長 (20300808) ; 長田 裕之 理化学研究所, 抗生物質研究室, 主任研究員 (80160836)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥100,500,000 (Direct Cost: ¥100,500,000); Fiscal Year 2004: ¥20,800,000 (Direct Cost: ¥20,800,000); Fiscal Year 2003: ¥20,300,000 (Direct Cost: ¥20,300,000); Fiscal Year 2002: ¥20,000,000 (Direct Cost: ¥20,000,000); Fiscal Year 2001: ¥20,400,000 (Direct Cost: ¥20,400,000); Fiscal Year 2000: ¥19,000,000 (Direct Cost: ¥19,000,000)
• Keywords: ヘパラナーゼ / VEGF / 血管新生 / がん細胞 / 化学生物学 / アクチン / Hsp60 / 血管新生阻害剤 / 分子標的 / PP2A / Bcl-2 / アポトーシス / 転移 / 血管内皮細胞遊走阻害 / 血管内皮細胞増殖阻害 / がん-間質相互作用 / PP2A阻害剤 / 前立腺がん / アンドロゲン受容体 / p53 / p21WAF1 / 乳がん / PTHrP(副甲状腺類縁タンパク) / 破骨細胞 / 骨転移 / 間質細胞 / Cuvularol / NK細胞活性化 / 15-deoxyspergualin / 癌増悪化 / 上皮-間質細胞間反応

Research Abstract

新規抗がん剤の探索研究として、血管内皮細胞増殖因子(VEGF)の作用を阻害する抗血管新生物質の探索を行い、epoxytwinol Aや新規フマギリン誘導体などの新規化合物を見出した。また、ヘパラン硫酸を模倣する化合物の探索を行った結果、RK-682から合成展開した4-Bn-RK682を高い特異性を有する抗ヘパラナーゼ物質として得た。さらに、がん細胞と間質細胞との共培養系によるがん細胞増殖阻害物質の探索を行ったところ、カビの培養液から一連のthielavin類を得た。
抗がん剤の化学生物学的手法による分子標的研究として、発見した化合物をビオチンや安定同位体で標識し結合タンパク質を同定した結果、amphidinolide Hはアクチンの200番目のチロシン残基に、またepolactaeneはHsp60の442番目のシステイン残基にそれぞれ細胞内で結合していることが示された。さらに、低分子化合物を官能基非依存的にビーズに固定化する技術を確立し、化合物に結合している内在性のタンパク質をマススペクトルにより同定することに成功した。
動物実験系での抗がん剤標的研究としては、ヘパラナーゼ阻害剤を検討した。ヘパラナーゼ阻害剤の4-Bn-RK-682およびKI-105はin vitroにおいてがん細胞の遊走・浸潤を阻害するだけではなく、in vivoにおいてもマウス腹腔内から注射したがん細胞の肺転移を経口投与で抑制した。

Research Products

• [Journal Article] Epolactaene binds human Hsp60 Cys^<442> resulting in the inhibition of chaperone activity. (2005)
  • Author(s): Nagumo, Y. et al.
  • Journal Title: Biochem.J. (in press)
• [Journal Article] Total synthesis of epoxyquinols A, B, and C and epoxytwinol A and the reactivity of a 2H-pyran derivative as the diene component in the diels-alder reaction. (2005)
  • Author(s): Shoji, M. et al.
  • Journal Title: J.Org.Chem. 70 Pages: 79-91
• [Journal Article] Apolipoprotein E3 (apoE3) safeguards pig proximal tubular LLC-PK1 cells against reduction in SGLT1 activity induced by gentamicin C. (2005)
  • Author(s): Takamoto, K. et al.
  • Journal Title: Biochim.Biophys. Acta (in press)
• [Journal Article] Involvement of doxorubicin-induced fas-expression in the antitumor effect of doxorubicin on lewis lung carcinoma in vivo. (2005)
  • Author(s): Yoshimoto, Y. et al.
  • Journal Title: Int.Immunopharmacol. 5 Pages: 281-288
• [Journal Article] Synthesis of a biotin-conjugate of phosmidosine O-ethyl ester as a G1 arrest antitumor drug. (2004)
  • Author(s): Sekine, M. et al.
  • Journal Title: Bioorg.Med.Chem. 12 Pages: 6343-6349
• [Journal Article] Structure-activity relationship of phosmidosine : importance of the 7,8-dihydro-8-oxoadenosine residue for antitumor activity. (2004)
  • Author(s): Sekine, M. et al.
  • Journal Title: Bioorg.Med.Chem. 12 Pages: 5193-5201
• [Journal Article] A new VHR dual-specificity protein tyrosine phosphatase inhibitor from dendrobium moniliforme. (2004)
  • Author(s): Bae, EY. et al.
  • Journal Title: Planta Med. 70 Pages: 869-870
• [Journal Article] Exip, a splicing variant of p38α, participates in interleukin-1 receptor proximal complex and downregulates NF-kappaB pathway. (2004)
  • Author(s): Yagasaki, Y. et al.
  • Journal Title: FEBS Lett. 575 Pages: 136-140
• [Journal Article] Kaurene diterpene induces apoptosis in human leukemia cells partly through a caspase-8-dependent pathway. (2004)
  • Author(s): Kondoh, M. et al.
  • Journal Title: Pharmacol Exp.Ther. 311 Pages: 115-122
• [Journal Article] Amphidinolide h, a potent cytotoxic macrolide, covalently binds on actin subdomain 4 and stabilizes actin filament. (2004)
  • Author(s): Usui, T. et al.
  • Journal Title: Chem.Biol. 11 Pages: 1269-1277
• [Journal Article] Structure-based design of a selective heparanase inhibitor as an antimetastatic agent. (2004)
  • Author(s): Ishida, K. et al.
  • Journal Title: Mol. Cancer Ther. 3 Pages: 1069-1077
• [Journal Article] Structure-activity relationships of epolactaene derivatives : structural requirements for inhibition of Hsp60 chaperone activity. (2004)
  • Author(s): Nagumo, Y. et al.
  • Journal Title: Bioorg.Med.Chem. 14 Pages: 4425-4429
• [Journal Article] RK-805,an endothelial-cell-growth inhibitor produced by Neosartorya sp., and a docking model with methionine aminopeptidase-2. (2004)
  • Author(s): Asami, Y. et al.
  • Journal Title: Tetrahedron. 60 Pages: 7085-7091
• [Journal Article] Heparanase as a molecular target of cancer chemotherapy. (2004)
  • Author(s): Simizu, S. et al.
  • Journal Title: Cancer Sci. 95 Pages: 553-558
• [Journal Article] The phosphorylation status and anti-apoptotic activity of Bcl-2 are regulated by ERK and protein phosphatase 2A on the mitochondria. (2004)
  • Author(s): Tamura, Y. et al.
  • Journal Title: FEBS Lett. 569 Pages: 249-255
• [Journal Article] The anti-cancer natural product pironetin selectively targets Lys352 of alpha-tubulin. (2004)
  • Author(s): Usui, T. et al.
  • Journal Title: Chem.Biol. 11 Pages: 799-806
• [Journal Article] Rational design and synthesis of novel heparan sulfate mimetic compounds as antiadhesive agents. (2004)
  • Author(s): Ishida, K. et al.
  • Journal Title: Bioorg.Med.Chem.Lett. 14 Pages: 2505-2509
• [Journal Article] ECH, an epoxycyclohexenone derivative that specifically inhibits Fas ligand-dependent apoptosis in CTL-mediated cytotoxicity. (2004)
  • Author(s): Mitsui.T.et al.
  • Journal Title: J.Immunol. 172 Pages: 3428-3436
• [Journal Article] Novel heparan sulfate mimetic compounds as antitumor agents. (2004)
  • Author(s): Ishida, K. et al.
  • Journal Title: Chem. & Biol. 11 Pages: 367-377
• [Journal Article] Dephosphorylation of Bcl-2 by protein phasphatase 2A resulted in apoptosis resistance. (2004)
  • Author(s): Simizu, S. et al.
  • Journal Title: Cancer Sci. 95 Pages: 266-270
• [Journal Article] Different reaction modes for the oxidative dimerization of epoxyquinols and epoxyquinones. importance of intermolecular hydrogen-bonding. (2004)
  • Author(s): Shoji, M. et al.
  • Journal Title: J.Org.Chem. 69 Pages: 1548-1556
• [Journal Article] Exploitation of heparanase inhibitors from microbial metabolites using an efficient visual screening system. (2004)
  • Author(s): Ishida, K. et al.
  • Journal Title: J.Antibiot. 57 Pages: 136-142
• [Journal Article] Synthesis of chemically stabilized phosmidosine analogues and the structure-activity relationship of phosmidosine. (2004)
  • Author(s): Sekine, M. et al.
  • Journal Title: J.Org.Chem. 69 Pages: 314-326
• [Journal Article] ICM0301s, new angiogenesis inhibitors from Aspergillus sp. F-1491. I. Taxonomy, fermentation, isolation and biological activities. (2004)
  • Author(s): Kumagai, H. et al.
  • Journal Title: J. Antibiotics 57 Pages: 97-103
• [Journal Article] ICM0301s, new angiogenesis inhibitors from Aspergillus sp. F-1491. II. Physico-chemical properities and structure elucidation. (2004)
  • Author(s): Someno, T. et al.
  • Journal Title: J. Antibiotics 57 Pages: 104-109
• [Journal Article] PP2A inhibitors, harzianic acid and related compounds produced by fungus strain F-1531. (2004)
  • Author(s): Kawada, M. et al.
  • Journal Title: J. Antibiotics 57 Pages: 235-237
  • NAID: 10012685872
• [Journal Article] A microplate assay for selective measurement of growth of epithelial tumor cells in direct coculture with stromal cells. (2004)
  • Author(s): Kawada, M. et al.
  • Journal Title: Anticancer Res. 24 Pages: 1561-1568
• [Publications] Watanabe, N.et al.: "M-phase kinases induce phospho-dependent ubiquitination of somatic Weel by SCFbTrCP."Proc.Natl.Acad.Sci., USA.. (in press). (2004)
• [Publications] Minamiguchi, K.et al.: "Ectopic expression of the amino-terminal peptide of androgen receptro leads to androgen receptro dysfunction and inhibition of androgen receptor-mediated prostate cancer growth."Mol.Cell.Endocrinol.. (in press). (2004)
• [Publications] Simizu, S.et al.: "Secretion of heparanase protein is regulated by glycosylation in human tumor cell lines."J.Biol.Chem.. 279. 2697-2703 (2004)
• [Publications] Woehlecke, H.et al.: "Reversal of breast cancer resistance protein-mediated drug resistance by tryprostatin A."Int.J.Cancer.. 107. 721-728 (2003)
• [Publications] Ganesh, S.et al.: "The lafora disease gene product laforin interacts with HIRIP5, a phylogenetically conserved protein containing a Nifu-like domain."Hum.Mol.Genet.. 12. 2359-2368 (2003)
• [Publications] Kanoh, N.et al.: "Immobilization of natural products on glass slides by using a photoaffinity reaction and the detection of protein-small molecule interactions."Angew.Chem.Int.Ed.. 42. 5584-5587 (2003)
• [Publications] Kakeya, H.et al.: "Novel non-peptide inhibitors targeting death receptor-mediated apoptosis."Bioorg.Med.Chem.Lett.. 13. 3743-3746 (2003)
• [Publications] Su, S.et al.: "Synthesis and cell cycle inhibition of the peptide enamide natural products terpeptin and the aspergillamides."Tetrahedron. 59. 8931-8946 (2003)
• [Publications] Maruyama, M.et al.: "Renal abnormalities in mice Caused by insufficiency of p38 α."J.Rec.Sig.Trans.. 23. 173-183 (2003)
• [Publications] Shoji, M.et al.: "Reaction modes of oxidative dimerization of epoxycyclohexenols."Tetrahedron Lett.. 44. 7205-7207 (2003)
• [Publications] Palaniappan, N.et al.: "Enhancement and selective production of phoslactomycin B, a protein phosphatase iia inhibitor, through identification and engineering of the corresponding biosynthetic gene cluster."Planta Medica. 69. 377-379 (2003)
• [Publications] Sekiyama, Y.et al.: "Biosynthesis of phoslactomycins : cyclohexanecarboxylic acid as the starter unit."Tetrahedron. 59. 7465-7471 (2003)
• [Publications] Nagashima, F.et al.: "Apoptosis-inducing properties of ent-kaurene-type diterpenoids from the liverwort jungermannia truncat α."Planta Medica. 69. 377-379 (2003)
• [Publications] Hayashi, Y.et al.: "Asymmetric total synthesis of pseurotin A."Organic Lett.. 5. 2287-2290 (2003)
• [Publications] Simizu, S.et al.: "Expression of heparanase in human tumor cell lines and human head and neck tumors."Cancer Lett.. 193. 83-89 (2003)
• [Publications] Kawatani, M.et al.: "Transmembrane domain of Bcl-2 is required for inhibition of ceramide synthesis, but not cytochrome c release in the pathway of inostamysin-induced apoptosis."Exp.Cell.Res.. 286. 57-66 (2003)
• [Publications] Osada, H.: "Development and application of bioprobes for mammalian cell cycle analyses."Cur.Med.Chem.. 10. 727-732 (2003)
• [Publications] Miyake, Y.et al.: "Epoxycyclohexenone inhibits fas-mediated apoptosis by blocking activation of pro-caspase-8 in the death-inducing signaling complex."J.Biol.Chem.. 278. 11213-11220 (2003)
• [Publications] Nakazawa, J.et al.: "A novel action of terpendole E on the motor activity of mitotic kinesin Eg5."Chem.& Biol.. 10. 131-137 (2003)
• [Publications] Nakamura, M.et al.: "Nordihydroguaiaretic acid, of a new family of microtubule-stabilizing agents, shows effects differentiated from paclitaxel."Biosci.Biotechnol.Biochem.. 67. 151-157 (2003)
• [Publications] Takamoto, K.et al.: "Aminoglycoside antibiotics reduce glucose reabsorption in kidney through down-regulation of SGLT1."Biochem.Biophys.Res.Commun.. 308. 866-871 (2003)
• [Publications] Kawada, M., Ishizuka, M.: "Inhibition of CTP : Phosphocholine cytidylyltransferase activity by 15-deoxyspergualin."J.Antibiot.. 56. 725-726 (2003)
• [Publications] Minamiguchi, K.et al.: "Androgen-independent prostate cancer DU-145 cells suppress androgen-dependent growth of prostate stromal cells through production of inhibitory factors for androgen responsiveness."Biochem.Biophys.Res.Commun.. 306. 629-636 (2003)
• [Publications] Kawada, M.et al.: "Specific inhibitors of protein phosphatase 2A inhibit tumor metastasis through augmentation of natural killer cells."Int.Immunopharmacol.. 3. 179-188 (2003)
• [Publications] M.Kawada: "Specific inhibitors of protein phosphatase 2A inhibit tumor metastasis through augmentation of natural killer cells"Int.Immunopharmacol.. 3. 179-188 (2003)
• [Publications] M.Kawada: "Hygrolidin induces p21 expression and abrogates cell cycle progression at G1 and S phases"Biochem.Biophys.Res.Commun.. 298. 178-183 (2002)
• [Publications] M.Kawada: "15-deoxyspergualin inhibits Akt kinase activation and phosphatidylcholine synthesis"J.Biol.Chem.. 277. 27765-27771 (2002)
• [Publications] M.Shintani: "Overexpression of cyclin D1 contributes to malignant properties of esophageal tumor cells by increasing VEGF production and decreasing Fas expression"Anticancer Res.. 22. 639-647 (2002)
• [Publications] K.Machida: "A novel adenine nucleotide translocase inhibitor, MT-21, induces cytochrome C release by a mitochondrial permeability transition-independent mechanism"J.Biol.Chem.. 277. 31243-31248 (2002)
• [Publications] Y.Miyamoto: "Identification of Saccharomyces cerevisiae isoleucyl-tRNA synthetase as target of G1-specific inhibitor reveromycin A"J.Biol.Chem.. 277. 28810-28814 (2002)
• [Publications] A.Matsuyama: "In vivo destabilization of dynamic microtubules by HDAC6-mediated deacetylation"EMBO J.. 24. 6820-6831 (2002)
• [Publications] Y.Hayashi: "Asymmetric total synthesis of (-)-azaspirene, a nevel angiogenesis inhibitor"J.Am.Chem.Soc.. 124. 12078-12079 (2002)
• [Publications] H.Kakeya: "Epoxyquinol A, a highly functionalized pentaketide dimmer with antiangiogenic activity isolated from fungal metabolites"J.Am.Chem.Soc.. 124. 3496-3497 (2002)
• [Publications] M.Shoji: "Total synthesis of (+)-epoxyquinols A and B"Angew.Chem.Int.Ed.. 41. 3192-3194 (2002)
• [Publications] Y.Asami: "Azaspirene : a novel angiogenesis inhibitor containing a 1-oxa-7-azaspiro [4.4]non-2-ene-4,6-dione skeleton produced by a fungi Neosartorya sp."Orgainc Lett.. 4. 2845-2848 (2002)
• [Publications] K.Ueda: "4-isoavenaciolide covalently binds and inhibits VHR, a dual-specificity phosphatase"FEBS Lett.. 525. 48-52 (2002)
• [Publications] N.Takahashi-Ando: "A novel lactonohydrolase responsible the detoxification of zearalenone. -Enzyme purification and gene cloning"Biochem.J.. 365. 1-6 (2002)
• [Publications] Iijima, M.: "Inhibition of ribonucleotide reductase activity by IC202C"J.Antibiot.. 54. 682-683 (2001)
• [Publications] Minamiguchi, K: "Thiolutin, an inhibitor of huvec adhesion to bitronectin, reduces paxilin in huvecs and suppresses tumor cell-induced angiogenesis"Int.J.Cancer. 93. 307-316 (2001)
• [Publications] Matsumoto, N.: "Synthesis and biological evaluation of cytogenin derivatives"J.Antibiot.. 54. 285-296 (2001)
• [Publications] Hamaguchi, T.: "TU-572, a potent and selective CD45 inhibitor, suppresses IgE-mediated anaphylaxis and murine contact hypersensitivity reactions"Int.Arch.Allergy Immunol.. 126. 318-324 (2001)
• [Publications] Kakeya, H.: "Lucilactaene, a new cell cycle inhibitor in p53-transfected cancer cells, produced by Fusarium sp"J.Antibiot.. 54. 850-854 (2001)
• [Publications] Nie, L.: "A facile and effective screening method for p21WAF1 promoter activator from microbial metabolites"J.Antibiot.. 54. 783-788 (2001)
• [Publications] Osada, H.: "An overview on the diversity of actinomycete metabolites"Actinomycetol.. 15. 11-14 (2001)
• [Publications] Sano, T.: "Isolation of new protein phosphatase inhibitors from two Cyanobacteria species, Planktothrix sp"J.Nat.Prod.. 64. 1052-1055 (2001)
• [Publications] Sodeoka, M.: "Synthesis of a tetronic acid library focused on inhibitors of tyrosine and dual-specificity protein phosphatases and its evaluation regarding VHR and Cdc25B inhibition"J.Med.Chem.. 44. 3216-3222 (2001)
• [Publications] Togashi, K.: "Inhibition of telomerase activity by fungus metabolites, CRM646-A and thielavin B"Biosci.Biotechnol.Biochem.. 65. 651-653 (2001)
• [Publications] Honda, Y.: "Isolation, and biological properties of a new cell cycle inhibitor, curvularol, isolated from Curvularia sp.RK97-F166"J.Antibiot.. 54. 10-16 (2001)
• [Publications] Ueki, M.: "A new trichostatin derivative, trichostatin RK, from Streptomyces sp.RK98-A74"J.Antibiot.. 54. 1093-1095 (2001)
• [Publications] Usui, T.: "Design and Synthesis of a Dimeric Derivative of RK-682 with Increased Inhibitory Activity against VHR, a Dual-Specificity ERK Phosphatase"Chemistry & Biology. 8. 1209-1220 (2001)
• [Publications] Kawada,M.: "The long-lasting antiproliferative effect of 15-deoxyspergulain through its spermidine moiety."J.Antibiot.. 53. 705-710 (2000)
• [Publications] Iijima,M.: "Biosynthesis of IC202C, a new siderophore with immunosupressive activity."J.Antibiot.. 53. 1411-1415 (2000)
• [Publications] Sudo,T.: "p62 functions as a p38 MAP kinase regulatior."Biochem.Biophys.Res.Commun.. 269. 521-525 (2000)
• [Publications] Ganesh,S.: "Laforin, defective in the progressive myoclonus epilepsy of Lafora type, is a dual-specificity phosphatase associated with polyribosomes."Human Mol.Genet.. 9. 2251-2261 (2000)
• [Publications] Ko,H.R.: "CRM646-A and -B, novel fungal metabolites that inhibit heparinase."J.Antibiot.. 53. 211-214 (2000)
• [Publications] Watanabe,H.: "Synthesis of pironetin and related analogs : Studies on structure-activity relationships as tubulin assembly inhibitors."J.Antibiot.. 53. 540-545 (2000)
• [Publications] Watabe,M.: "Activation of MST/Krs and c-Jun N-terminal kinases by different signaling pathways during cytotrienin A-induced apoptosis."J.Biol.Chem.. 275. 8766-8771 (2000)
• [Publications] Matsumoto,M.: "Involvement of p38 mitogen-activated protein kinase signaling pathway in osteoclastogenesis mediated by receptor activator of NF-κ BLigand(RANKL)."J.Biol.Chem.. 275. 31155-31161 (2000)
• [Publications] Wang,H.: "Synthesis and evaluation of tryprostatin B and demethoxyfumitremorgin C analogs."J.Med.Chem.. 43. 1577-1585 (2000)
• [Publications] Simizu,S.: "Bcl-2 inhibits calcineurin-mediated Fas ligand expression in antitumor drug-induced baby hamster kidney cells."Jpn.J.Cancer Res.. 91. 706-714 (2000)
• [Publications] Simizu,S.: "Mutations of Plk gene lead to instability of Plk protein in human cancer cell lines."Nature Cell Biol.. 2. 852-854 (2000)
• [Publications] Honda,Y.: "Isolation, and biological properties of a new cell cycle inhibitor, curvularol, isolated from Curvularia sp.RK97-F166."J.Antibiot.. 54. 10-16 (2001)