1998 Fiscal Year Annual Research Report

DNAプライマーゼ及びテロメラーゼを分子標的としたがん細胞増殖抑制

Research Project

Project/Area Number	10153224
Research Institution	Nagoya University
Principal Investigator	吉田松年名古屋大学, 医学部, 教授 (70090420)
Co-Investigator(Kenkyū-buntansha)	村手隆名古屋大学, 医学部, 助教授 (30239537) 鈴木元名古屋大学, 医学部, 講師 (80236017) 小泉恵子名古屋大学, 医学部, 助教授 (00118027)
Keywords	プライマーゼ / スフィンゴシン / アポトーシス / テロメラーゼ / 阻害剤 / イソチアゾロン
Research Abstract	1) 目的従来、制癌剤の分子標的として注目されなかったが重要な機能を果たしているプライマーゼ及びテロメラーゼを標的として、新しい制がん剤を開発する事を本研究の目的とする。 2) プライマーゼ我々はスフィンゴシンが強力にプライマーゼを阻害する事を見い出した(Simbulanら,Biochemistry 33:9007,1994).そこでスフィンゴシン異性体、セラミド等関連化合物9種類によるプライマーゼ活性の阻害を試験管内で検討した所、天然スフィンゴシン(trans型)が最も阻害作用が強く、phytoshingosine.N,N-dimethylshingosineがそれに続き、セラミドやスフィンゴミエリンは阻害しなかった。培養ヒト白血病細胞に対してそれらの増殖抑制・アポトーシス誘導を調べたところ、プライマーゼ阻害と増殖抑制とは良い相関を示した(T-Koizumiら,Biochem.Mol.Biol.Int.41:1179,1997)。又、スフィンゴシンをリード化合物として種々の誘導体を化学合成し、構造活性相関を明らかにした(未発表)。 3) テロメラーゼ阻害剤:ラット腹水肝がん細胞より数段階のカラムクロマトグラフィーで高度に精製したテロメラーゼを用いて阻害剤のスクリーニングを行った。多種類の合成化合物の中から、強いテロメラーゼ阻害を示す化合物6種類を得た。その中の4種類はイソチアゾロンの誘導体であった。最も強く阻害活性を示した2-[3-(trifluoromethyl)phenyl]isothiazolin-3-one(TMPI)について阻害機構を調べた。テロメラーゼ活性測定系としては改良TRAP法を用いた。TMPIはテロメア配列を有する一次プライマーと非競合的に、またdNTPとほぼ競合的(mixed type)に反応を阻害した。Ki値は2.5μMであった(論文投稿中)。本化合物の培養細胞にたいする影響を検討している。

Research Products

(15 results)

All Other

All Publications (15 results)

[Publications] Horie Tadashi: "A5'-monophosphate form of bredinin selectively inhibits the activities of mammalian DNA polymerases in vitro." International Journal of Molecular Medicine. 1. 83-90 (1998)
[Publications] Mizushina Yoshiyuki: "Studies on inhibitors of mammalian DNA polymerase α and β." Biochemical Pharmacology. 55. 537-541 (1998)
[Publications] Tomikawa Aki: "Synthetic nucleotides and uncleosides. 40. selective inhibition of eukaryotic DNA polymerase α by 9-(β-D-arabinofuranosyl)-2-(p-n-butylanilino)adenine 5'-triphosphate(BuAara ATP)and its 2'-up azide analog:synthesis and enzymatic evaluations." Nucleosides and Nucleotides. 17. 487-501 (1998)
[Publications] Kato Osamu: "Serum DNA polymerase β as an indicator for fatal liver injury of rat induced by D-galactosamine hydrochloride and lipopolysaccharide." Biochimica et Biophysica Acta. 1380. 369-376 (1998)
[Publications] Nakayabu Mikiya: "Mismatched nucleosides may facilitate expansion of trinucleotide repeats in genetic diseases." Nucleic Acids Research. 26. 1980-1984 (1998)
[Publications] Mizushina Yoshiyuki: "An ergosterol peroxide,a natural product that selectivity enhances the inhibitory effect of linoleic acid on DNA polymerase β." Biol.Pharm.Bull.21. 444-448 (1998)
[Publications] Mizushina Yoshiyuki: "The inhibitory effect of novel trierpenoid compounds,fomitellic acids,on DNA polymerase β." Biochem.J.330. 1325-1332 (1998)
[Publications] Wan-Qian Zhao: "Cell cycle-associated accumulation of tissue inhibitor of metalloproteinases-1(TIMP-1)in the nuclei of human gingival fibroblasts." Journal of Cell Science. 111. 1147-1153 (1998)
[Publications] Yamaguchi Yosuke: "Change in telomerase activity of rat organs during growth and aging." Experimental Cell Research. 242. 120-127 (1998)
[Publications] Ohta Keisuke: "Sulfoquinovosyldiacylglycerol,KM043,a new poten inhibitor of eukaryotic DNA polymerases and HIV-reverse transcriptase type I from a marine red alga,Glgatina tenella." Chem.Pharm.Bull.46. 684-686 (1998)
[Publications] Ogawa Akio: "Lithocholic acid,a putative tumor promoter,inhibits mammalian DNA polymerase β." Japanese Journal of Cancer Research. 89. 1154-1159 (1998)
[Publications] Mizushina Yoshiyuki: "A mushroom fruiting body-inducing substance inhibits activities of replicative DNA polymerases." Biochem,Biophys.Res.Commun.249. 17-22 (1998)
[Publications] Togashi Hideaki: "4-hydroxy-17-methylincisterol,and inhibitor of DNA polymerase-a activity and the growth of human cancer cells in vitro." Biochem.Pharmacol.56. 583-590 (1998)
[Publications] Isogai Chiho: "Analysis of bax protein in sphingosine-induced apoptosis in the human leukemic cell line TF1 and its bcl-2 transfectants." Exp.Hematol.26. 1118-1125 (1998)
[Publications] Ogawa Akio: "Sulfated glycoglycerolipid from archaebacterium inhibits eukaryotic DNA polymerase α,β,and retroviral reverse transcriptase and affects methyl methanesulfonate cytotoxicity." Int.J.Cancer. 76. 512-518 (1998)

1998 Fiscal Year Annual Research Report

DNAプライマーゼ及びテロメラーゼを分子標的としたがん細胞増殖抑制

Principal Investigator

吉田 松年 名古屋大学, 医学部, 教授 (70090420)

Research Products

[Publications] Horie Tadashi: "A5'-monophosphate form of bredinin selectively inhibits the activities of mammalian DNA polymerases in vitro." International Journal of Molecular Medicine. 1. 83-90 (1998)

[Publications] Mizushina Yoshiyuki: "Studies on inhibitors of mammalian DNA polymerase α and β." Biochemical Pharmacology. 55. 537-541 (1998)

[Publications] Kato Osamu: "Serum DNA polymerase β as an indicator for fatal liver injury of rat induced by D-galactosamine hydrochloride and lipopolysaccharide." Biochimica et Biophysica Acta. 1380. 369-376 (1998)

[Publications] Nakayabu Mikiya: "Mismatched nucleosides may facilitate expansion of trinucleotide repeats in genetic diseases." Nucleic Acids Research. 26. 1980-1984 (1998)

[Publications] Mizushina Yoshiyuki: "An ergosterol peroxide,a natural product that selectivity enhances the inhibitory effect of linoleic acid on DNA polymerase β." Biol.Pharm.Bull.21. 444-448 (1998)

[Publications] Mizushina Yoshiyuki: "The inhibitory effect of novel trierpenoid compounds,fomitellic acids,on DNA polymerase β." Biochem.J.330. 1325-1332 (1998)

[Publications] Wan-Qian Zhao: "Cell cycle-associated accumulation of tissue inhibitor of metalloproteinases-1(TIMP-1)in the nuclei of human gingival fibroblasts." Journal of Cell Science. 111. 1147-1153 (1998)

[Publications] Yamaguchi Yosuke: "Change in telomerase activity of rat organs during growth and aging." Experimental Cell Research. 242. 120-127 (1998)

[Publications] Ohta Keisuke: "Sulfoquinovosyldiacylglycerol,KM043,a new poten inhibitor of eukaryotic DNA polymerases and HIV-reverse transcriptase type I from a marine red alga,Glgatina tenella." Chem.Pharm.Bull.46. 684-686 (1998)

[Publications] Ogawa Akio: "Lithocholic acid,a putative tumor promoter,inhibits mammalian DNA polymerase β." Japanese Journal of Cancer Research. 89. 1154-1159 (1998)

[Publications] Mizushina Yoshiyuki: "A mushroom fruiting body-inducing substance inhibits activities of replicative DNA polymerases." Biochem,Biophys.Res.Commun.249. 17-22 (1998)

[Publications] Togashi Hideaki: "4-hydroxy-17-methylincisterol,and inhibitor of DNA polymerase-a activity and the growth of human cancer cells in vitro." Biochem.Pharmacol.56. 583-590 (1998)

[Publications] Isogai Chiho: "Analysis of bax protein in sphingosine-induced apoptosis in the human leukemic cell line TF1 and its bcl-2 transfectants." Exp.Hematol.26. 1118-1125 (1998)

[Publications] Ogawa Akio: "Sulfated glycoglycerolipid from archaebacterium inhibits eukaryotic DNA polymerase α,β,and retroviral reverse transcriptase and affects methyl methanesulfonate cytotoxicity." Int.J.Cancer. 76. 512-518 (1998)

吉田松年名古屋大学, 医学部, 教授 (70090420)