和 漢 医 薬 学 総 合 研 究 所 年 報 第 四 一 巻 二 〇 一 四 年 富 山 大 学

AnnualReport

Insti tuteofNaturalMedi ci ne Uni versi tyofToyama

Vol .41 2014

和漢医薬学総合研究所年報第四一巻二〇一四年富山大学

和漢医薬学総合研究所年報

富 山 大 学

第 41 ^巻 2014 ^年

和漢医薬学総合研究所年報

富 山 大 学

第 41 ^巻 2014 ^年

富 山 大 学

和漢医薬学総合研究所

ᐩ

ᐩᒣ኱Ꮫ࿴₎་⸆Ꮫ⥲ྜ◊✲ᡤᖺሗ ᕳ ᖺ Annual Report of Institute of Natural Medicine

University of Toyama Vol. 41, 2014

┠ḟ

ᕳ㢌ゝ

⥲ ㄝ

Wntࢩࢢࢼࣝఏ㐩࡟స⏝ࡍࡿኳ↛⏤᮶໬ྜ≀ࡢ᥈⣴

␜㖭୍ᩥ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 2

ྛ㒊㛛࣭㝃ᒓࢭࣥࢱ࣮ࡢάື࡜ᴗ⦼

㈨※㛤Ⓨ㒊㛛 ⏕⸆㈨※⛉Ꮫศ㔝࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭12 ኳ↛≀໬Ꮫศ㔝 ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭19

⑓ែไᚚ㒊㛛 」ྜ⸆≀⸆⌮Ꮫศ㔝࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭24

⑓ែ⏕໬Ꮫศ㔝࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭29 ᾘ໬⟶⏕⌮Ꮫศ㔝࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭36

⚄⤒ᶵ⬟Ꮫศ㔝 ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭42

ᰤ㣴௦ㅰᏛศ㔝 ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭46

⮫ᗋ฼⏝㒊㛛 ₎᪉デ᩿Ꮫศ㔝࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭48 ᐤ㝃㒊㛛 ࿴₎⸆〇๣㛤Ⓨศ㔝࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭55 ᣐⅬ஦ᴗ᥎㐍ᐊ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭60 㝃ᒓẸ᪘⸆≀◊✲ࢭࣥࢱ࣮

ᅜ㝿ඹྠ◊✲ศ㔝࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭62 Ẹ᪘⸆≀㈨ᩱ㤋࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭65 ࢸࢽࣗ࢔ࢺࣛࢵࢡ᥎㐍≉༊

ኳ↛≀๰⸆ศ㔝࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭69

2014ᖺࡢάືグ㘓࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭73

⾲⣬ࡢ෗┿

࢘ࢥࢠ⛉ࡢP. vietnamensis Ha et Grushv. var. fuscidiscus K. Komatsu, S. Zhu & S. Q. Cai㸪㔝୕୐㸪majonoside R2

㸦୰ᅜ㞼༡┬༡㒊࡟࡚㸪2012ᖺ9᭶8᪥㸪ᑠᯇ࠿ࡘᏊ᧜ᙳ㸧

1999ᖺࡢᏛ⾡ㄪᰝ࡛ᮏ᳜≀ࢆⓎぢࡋ㸪እ㒊ᙧែ㸪㑇ఏᏊᆺཬࡧᡂศࡢ◊✲࡟ࡼࡾ㸪P. vietnamensisࡢ᪂ኚ✀࡜ࡋ࡚࿨ྡ

ࡋࡓࠋ࣋ࢺࢼ࣒ேཧ࡜ྠᵝ࡟࢜ࢥࢳ࣮ࣟࣝ⣔ࢧ࣏ࢽࣥࡢmajonoside R2ࢆྵ᭷ࡍࡿࡇ࡜ࡀ≉ᚩ࡛㸪ࡉࡽ࡟ginsenoside Rb1

ࡸRg1࡞࡝ࡶྵ᭷ࡋ࡚࠸ࡓࠋ㔝୕୐ࡢྡ⛠࡛⿵๣㸦ᙉኊ⸆㸧࡜ࡋ࡚ᕷሙ࡛኎ࡽࢀ࡚࠸ࡓࡀ㸪ᙜ᫬࠿ࡽ㔝⏕ရࡢῶᑡࡀ༴᝹

ࡉࢀ࡚࠾ࡾ㸪ᮧேࡢᡭ࡟ࡼࡾ᱂ᇵ໬ࡀᅗࡽࢀ࡚࠸ࡓࠋ2012ᖺ࡟෌ᗘゼၥࡋࡓ࡜ࡁ࡟ࡣᕷሙ࡟ࡣ඲ࡃぢࡽࢀࡎ㸪᱂ᇵࡶ⣽ࠎ

࡜⾜ࢃࢀ࡚࠸ࡿ⛬ᗘ࡛࠶ࡗࡓࠋ⸆⏝㈨※᳜≀ࡢ᭷⏝ᛶࡢホ౯࡜࡜ࡶ࡟㸪ᣢ⥆ྍ⬟࡞฼⏝ࡢࡓࡵࡢ᪉⟇ࢆ❧࡚ࡿࡇ࡜ࡀᛴົ

࡛࠶ࡿࠋ

◊

◊ ✲ ᡤ ᖺ ሗ ᕳ㢌 ࡢ ゝ ⴥ

࿴₎་⸆Ꮫ⥲ྜ◊✲ᡤࡣ1963ᖺ4᭶࡟ᐩᒣ኱Ꮫ⸆Ꮫ㒊㝃ᒓ࿴₎⸆◊✲᪋タ࡜ࡋ࡚タ⨨ࡉ

ࢀ㸪ࡑࡢ౑࿨ࡣ㸪⤒㦂▱ࡢ㞟✚࡛࠶ࡿఏ⤫་Ꮫ㸪࠶ࡿ࠸ࡣࡑࡇ࡛౑⏝ࡉࢀࡿ⸆≀࡟ࡘ࠸࡚㸪 ඛ➃⛉Ꮫᢏ⾡ࢆ㥑౑ࡋ࡚⛉Ꮫⓗ࡟ホ౯ࡋ࿴₎་⸆Ꮫ࡜すὒ་⸆Ꮫࡢ⼥ྜࢆࡣ࠿ࡾ㸪᪂ࡋ࠸

་⸆Ꮫయ⣔ࡢᵓ⠏࡜⮬↛⎔ቃࡢಖ඲ࢆྵࡵࡓ඲ே་⒪ࡢ☜❧࡟㈉⊩ࡍࡿࡇ࡜࡟࠶ࡾࡲࡍࠋ ᮏ◊✲ᡤࡣࡇࡢ౑࿨ࢆᯝࡓࡍ࡭ࡃ㸪≉Ⰽ࠶ࡿᏛ⾡◊✲ࣉࣟࢪ࢙ࢡࢺ࡟ྲྀࡾ⤌ࡳ㸪2010ᖺ

ࡼࡾ࿴₎་⸆Ꮫ࡟≉໬ࡋࡓᅜෆ၏୍ࡢ㝃⨨◊✲ᡤ࡜ࡋ࡚ᩥ㒊⛉Ꮫ኱⮧ࡼࡾࠕ࿴₎⸆ࡢ⛉Ꮫ ᇶ┙ᙧᡂᣐⅬࠖ࡟ㄆᐃࡉࢀࡲࡋࡓࠋᮏ஦ᴗ࡛ࡣ㸪௚ࡢ኱Ꮫࡸ◊✲ᶵ㛵ࡢ◊✲⪅࡜㐃ᦠࡋ࡚

࿴₎་⸆Ꮫ࡟㛵ࡍࡿබເᆺඹྠ◊✲ࢆ✚ᴟⓗ࡟㐍ࡵ㸪ᮏศ㔝ࡢⓎᒎ࡟㈉⊩ࡍࡿ◊✲ࢆᐇ᪋

ࡋ࡚ࡁࡲࡋࡓࠋࡲࡓ㸪2013ᖺࡢᣐⅬ஦ᴗ୰㛫ホ౯࡟࠾ࡅࡿࠕ₎᪉⸆ࡢస⏝ᶵᗎ࡟㛵ࡍࡿす ὒ་Ꮫ࡜ࡢ⼥ྜࠖ࡜ࡢᣦ᦬࡟ᑐࡋ࡚ࡣ≉ᐃ◊✲ࠕ₎᪉⸆ࡀ᭷ࡍࡿ」㞧⣔ࡢゎᯒࠖࢆタᐃࡋ㸪

₎᪉⸆ࡸ⏕⸆ࡢస⏝ᶵᗎࡢゎ᫂ࢆࢸ࣮࣐࡜ࡋࡓ◊✲ࢆ᥎㐍ࡋ࡚࠾ࡾࡲࡍࠋ

ᮏ◊✲ᡤࡢ⤌⧊ࡣ㸪3㒊㛛(8ศ㔝)࡜1ᐤ㝃㒊㛛࠿ࡽ࡞ࡿ◊✲㒊㸪ᅜ㝿ඹྠ◊✲㒊࡜Ẹ᪘⸆

≀㈨ᩱ㤋࠿ࡽ࡞ࡿẸ᪘⸆≀◊✲ࢭࣥࢱ࣮㸪࠾ࡼࡧᣐⅬ஦ᴗ᥎㐍ᐊ࡟ࡼࡾᵓᡂࡉࢀ࡚࠸ࡲࡍࠋ ࡋ࠿ࡋ㸪ᣐⅬ஦ᴗࢆࡉࡽ࡟᥎㐍ࡍࡿࡓࡵ࡟ࡣ㸪ࡼࡾᶵ⬟ⓗ࡞⤌⧊యไࢆᵓ⠏࣭ᩚഛࡍࡿᚲ せࡀ࠶ࡾ㸪ࢫࢱࢵࣇᵓᡂࢆྵࡵࡓ⤌⧊ࡢぢ┤ࡋ࡜෌⦅ࢆᅗࡗ࡚࠸ࡲࡍࠋ

Ẹ᪘⸆≀㈨ᩱ㤋ࡣ඲ᅜ࡟㄂ࡿ ᐩᒣ኱Ꮫࢆ௦⾲ࡍࡿ᪋タ࡜ࡋ࡚㸪୰㧗⏕ࢆᑐ㇟࡜ࡋࡓࠕࡦ

ࡽࡵࡁ☆࡜ࡁࡵࡁࢧ࢖࢚ࣥࢫࠖ࡞࡝ࡢ୍⯡බ㛤஦ᴗࢆ✚ᴟⓗ࡟⾜࠺࡜࡜ࡶ࡟㸪ࢽ࣮ࣗࢫࣞ

ࢱ࣮ห⾜ࢆ⥅⥆ࡍࡿ࡞࡝㸪ࡼࡾᗈࡃⓎಙࡍࡿࡇ࡜࡟ດࡵ࡚࠾ࡾࡲࡍࠋ

ࡇࡢࡼ࠺࡟㸪ඹྠ฼⏝࣭ඹྠ◊✲ᣐⅬࡢ⤌⧊࣭ᶵ⬟ࡢ඘ᐇࢆᅗࡾࡘࡘ㸪ᡤဨࡀ୍୸࡜࡞

ࡾ࿴₎་⸆Ꮫ◊✲ࡢ㐍ᒎࢆᅗࡿᡤᏑ࡛ࡍࡢ࡛㸪௒ᚋඹ㸪ⓙᵝ᪉࠿ࡽࡢ୍ᒙࡢࡈᨭ᥼ࢆ㈷ࡾ

ࡲࡍࡼ࠺࠾㢪࠸⏦ࡋୖࡆࡲࡍࠋ

ᖹᡂ27ᖺ1᭶

࿴₎་⸆Ꮫ⥲ྜ◊✲ᡤ㻌 ᡤ㛗㻌 㻌ᰘ ཎ ┤ ฼

総 説

Wntࢩࢩࢢࢼࣝఏ㐩࡟స⏝ࡍࡿኳ↛⏤᮶໬ྜ≀ࡢ᥈⣴

ᐩᒣ኱Ꮫ࿴₎་⸆Ꮫ⥲ྜ◊✲ᡤ ㈨※㛤Ⓨ◊✲㒊㛛 ⏕⸆㈨※⛉Ꮫศ㔝

␜㖭୍ᩥ

1. ࡣࡌࡵ࡟

⮬↛⏺䛾Ꮡᅾ䛩䜛ከ✀ከᵝ䛺⏕≀✀䛿䠈䛭䛾⏕⫱⎔ቃ䜔㐍໬䛻ᛂ䛨䛶ᵝ䚻䛺஧ḟ௦ㅰ⏘≀䠈䛩䛺䜟䛱ኳ↛≀䜢

⏘⏕䛩䜛䛣䛸䛜▱䜙䜜䛶䛔䜛䠊ኳ↛≀䛿ே▱䛾ཬ䜀䛺䛔䝴䝙䞊䜽䛺໬Ꮫᵓ㐀䛸⯆࿡῝䛔⏕≀άᛶ䜢♧䛩䜒䛾䛜ከ䛟 ሗ࿌䛥䜜䛶䛚䜚䠈་⸆ရ䛾䝸䞊䝗໬ྜ≀䜔⏕࿨⌧㇟ゎ᫂䛾䛯䜑䛾◊✲⏝ヨ⸆䠄䜿䝭䜹䝹䝒䞊䝹䠅䛺䛹䛸䛧䛶ᛂ⏝䛥䜜䠈

ྛ✀⑌ᝈ἞⒪䜔⏕࿨⛉Ꮫ◊✲䛾᥎㐍䛻኱䛝䛟㈉⊩䛧䛶䛝䛯¹䠊

ᡃ䚻䛿䠈䛜䜣⑌ᝈ䛻㛵䜟䜛䝅䜾䝘䝹ఏ㐩䛻స⏝䛩䜛ኳ↛≀䛾Ⓨぢ䜢┠ᣦ䛧䠈䝇䜽䝸䞊䝙䞁䜾◊✲䜢⾜䛳䛶䛚䜚䠈᭷

⏝䛺⏕≀άᛶศᏊ䛾๰〇䜢┠ᣦ䛧䛶䛔䜛䠊ᮏ✏䛷䛿䛜䜣⑌ᝈ䜢䛿䛨䜑䠈ᵝ䚻䛺⑌ᝈ䜔⏕࿨⌧㇟䛻㛵୚䛩䜛䛣䛸䛜▱

䜙䜜䛶䛔䜛䜴䜲䞁䝖䠄Wnt䠅䝅䜾䝘䝹䛻╔┠䛧䛯⏕≀άᛶኳ↛≀䛾᥈⣴◊✲䛻䛴䛔䛶⤂௓䛩䜛䠊 2. Wntࢩࢢࢼࣝ࡜ࡣ

Wnt 䝅䜾䝘䝹䛿䠈⥺⹸䠈䝅䝵䜴䝆䝵䜴䝞䜶䠈䝊䝤䝷䝣䜱䝑䝅䝳䠈䝬䜴䝇䛚䜘䜃䝠䝖䛻⮳䜛䜎䛷ᵝ䚻䛺⏕≀✀䛻ಖᏑ䛥䜜䛶 䛔䜛䝅䜾䝘䝹ఏ㐩⤒㊰䛷䠈ึᮇⓎ⏕䛻䛚䛡䜛య㍈ᙧᡂ䛛䜙ྛ✀⤌⧊䞉ჾᐁ䛾ᙧែᙧᡂ䠈⣽⬊䛾ศ໬䞉ቑṪ䠈⤌⧊ᖿ

⣽⬊䛾⥔ᣢ䠈Ⓨ䛜䜣䛻䛚䛡䜛ᵝ䚻䛺⏕࿨⌧㇟䛾㐣⛬䛻䛚䛔䛶ከᙬ䛛䛴㔜せ䛺ᙺ๭䜢ᢸ䛳䛶䛔䜛䠊Wnt䛿ศᏊ㔞⣙4

୓䛾ศἪᆺ⢾䝍䞁䝟䜽䛷䛒䜚䠈့ங㢮䛻䛚䛔䛶䛿⌧ᅾ䜎䛷䛻19✀䛾Wnt䝣䜯䝭䝸䞊䛜▱䜙䜜䛶䛔䜛䠊ᮏ䝅䜾䝘䝹⤒㊰

䛿䠈1䠅β-catenin/TCF䠄T-cell factor䠅䜢௓䛧䛶ᶆⓗ㑇ఏᏊ䛾Ⓨ⌧䜢ㄪ⠇䛩䜛Wnt/β-catenin⤒㊰䠈2䠅⣽⬊㦵᱁⣔䛾ไ ᚚ䛻㛵୚䛩䜛⣽⣽⬊ෆᖹ㠃ᴟᛶ䠄PCP䠅⤒㊰䠈3䠅⣽⬊䛾᥋╔䠈㐠ື䛻㛵୚䛧䛶䛔䜛Wnt/Ca²⁺⤒㊰䛾୕䛴䛜▱䜙䜜䛶䛔 䜛䠊β-catenin/TCF ⤒㊰䛿ྂ඾ⓗ⤒㊰䛸䜒࿧䜀䜜 β-catenin 䛻౫Ꮡ䛩䜛䛜䠈௚䛾஧䛴䛿 β-catenin 䛻౫Ꮡ䛧䛺䛔䠊 β-catenin䛿cadherin⤖ྜ䝍䞁䝟䜽䛸䛧䛶ྠᐃ䛥䜜䠈⣽⬊᥋╔䛻䛚䛔䛶㔜せ䛺ᙺ๭䜢ᢸ䛳䛶䛔䜛୍᪉䛷䠈β-catenin/TCF

⤒㊰䛻䛚䛔䛶䛿䝯䝕䜱䜶䞊䝍䞊䛸䛧䛶ᶵ⬟䛧䠈Wnt 䝅䜾䝘䝹䛾ᶆⓗ㑇ఏᏊ䛾㌿෗ㄪ⠇䛻㛵䜟䜛䠊䛣䛾䛖䛱䠈ᮏ✏䛷䛿 β-catenin/TCF⤒㊰䛻╔┠䛧䛶䛔䜛䠊

Wnt䝸䜺䞁䝗䝍䞁䝟䜽䛜䛺䛔≧ែ䛩䛺 䜟䛱Wnt䝅䜾䝘䝹䛜OFF䛾≧ែ䛻 䛚䛔䛶䠈⣽⬊㉁ෆ䛾 β-catenin 䛿 casein kinase-1α䠄CK1α䠅䠈glycogen synthase kinase 3β䠄GSK3β䠅䠈Axin䠈 adenomatous polyposis coil䠄APC䠅䛺 䛹䛸」ྜయ䠄β-catenin ศゎ」ྜయ䠅 䜢ᙧᡂ䛧䠈2 䛴䛾䜻䝘䞊䝊 CK1α䠈

GSK3β 䛻䜘䜛䝸䞁㓟໬䜢ཷ䛡䠈䝥䝻

䝔䜰䝋䞊䝮⣔䛻䛚䛔䛶ศゎ䛥䜜䜛䠊

୍᪉ Wnt 䝸䜺䞁䝗䝍䞁䝟䜽䛜Ꮡᅾ䛩 䜛䠈䛩䛺䜟䛱Wnt䝅䜾䝘䝹䛜ON䛾

≧ ែ 䛻 䛚 䛔 䛶 䠈Wnt 䝍 䞁 䝟 䜽 䛜 ᅗ1㻌Wnt䝅䜾䝘䝹⤒㊰䜾 ㊰

frizzled䠄Fz䠅ཷᐜయ䛚䜘䜃ඹཷᐜయ䛷䛒䜛lipoprotein-related protein 5/6䠄LRP5/6䠅䛻⤖ྜ䛩䜛䛸䠈β-catenin䛾䝸䞁㓟໬䠈 䝥䝻䝔䜰䝋䞊䝮⣔䛻䜘䜛ศゎ䛜ᢚไ䛥䜜䠈⣽⬊ෆ䛻 β-catenin 䛜⵳✚䛥䜜䜛䠊⵳✚䛧䛯 β-catenin䛿䛭䛾ᚋ䠈᰾ෆ䜈⛣

⾜䛧TCF䛸」ྜయ䜢ᙧᡂ䛧䠈ᶆⓗ㑇ఏᏊ䛷䛒䜛c-myc䠈cyclin D1䠈PPARδ䠈COX-2䛺䛹䛾㌿෗䛜ஹ㐍䛥䜜䜛䠄ᅗ1䠅^2-4䠊 ᮏ䝅䜾䝘䝹䛿๓㏙䛾䜘䛖䛻ึᮇⓎ⏕䜔⤌⧊䞉ჾᐁᙧᡂ䛺䛹䛾㔜せ䛺⏕࿨⌧㇟䛻㛵䜟䛳䛶䛔䜛୍᪉䛷䠈኱⭠䛜䜣䜢 䛿䛨䜑䛸䛩䜛✀䚻䛾䛜䜣⑌ᝈ䛻䛚䛡䜛␗ᖖஹ㐍䛜ሗ࿌䛥䜜䛶䛔䜛䠊䜎䛯䠈⢾ᒀ⑓䜔⢭⚄⑌ᝈ䛺䛹ྛ✀⑌ᝈ䛸䛾㛵䜟䜚 䜒♧၀䛥䜜䛶䛔䜛䠊䛣䛾䜘䛖䛺⫼ᬒ䛛䜙Wnt䝅䜾䝘䝹䛻స⏝䛩䜛໬ྜ≀䛿་⸆ရ䝸䞊䝗໬ྜ≀䜔ྛ✀⏕࿨⌧㇟䜢◊✲

䛩䜛ୖ䛷䛾ศᏊ䝒䞊䝹䛸䛺䜛䛣䛸䛜ᮇᚅ䛥䜜䜛䠊 3. Wnt䝅䝅䜾䝘䝹䜢ᶆⓗ䛸䛧䛯䝇䜽䝸䞊䝙䞁䜾⁵

ᙜ◊✲ᐊ䛷⊂⮬䛻ᵓ⠏䛧䛯ኳ↛㈨※ᢳฟ≀䝷䜲䝤䝷䝸䞊䛾䛖䛱䠈䝞䞁䜾䝷䝕䝅䝳䛚䜘䜃䝍䜲䛻䛶᥇ྲྀ䛧䛯᳜≀ᢳฟ

≀䜢ᑐ㇟䛸䛧䛶䠈TOPFlash/FOPFlash䝹䝅䝣䜵䝷䞊䝊䜰䝑䝉䜲䝅䝇䝔䝮⁵䜢⏝䛔䛶Wnt䝅䜾䝘䝹䜢ホ౯䛧䛯䠊ᮏ䜰䝑䝉䜲 䝅䝇䝔䝮䛿䠈㔝⏕ᆺ TCF ⤖ྜ㡿ᇦ䠄CCTTTGATC䠅䜢䜒䛴 SuperTOPFlash 䝺䝫䞊䝍䞊㑇ఏᏊ䜢䝠䝖⫾ඣ⭈⣽⬊

HEK293䛻Ᏻᐃᑟධ䛧䛯STF/293⣽⬊䜢⏝䛔䛯䠊ヨᩱῧຍ䛻䜘䜛䝹䝅䝣䜵䝷䞊䝊άᛶ䠄TOPάᛶ䠅䛾ኚ໬䜢 ᐃ䛩䜛

䛣䛸䛻䜘䜚 Wnt 䝅䜾䝘䝹䛾᭱ୗὶ䛻఩⨨䛩䜛 TCF/β-catenin ㌿෗άᛶ䛾ホ౯䛜ྍ⬟䛷䛒䜛䠊ᮏ⣽⬊䛿ෆᅾᛶ β-catenin㔞䛜ప䛔䛯䜑䠈䛭䛾䜎䜎ヨ㦂䛻⏝䛔䜛䛸TOPάᛶ䛜ᑠ䛥䛔䠊䛭䛾䛯䜑䠈ヨᩱῧຍ䛾㝿GSK3β㜼ᐖ๣䛷䛒䜛 LiCl䜢䛒䜟䛫䛶ῧຍ䛩䜛䛣䛸䛻䜘䜚䠈β-catenin䛾ศゎ䜢ᢚไ䛧䠈TOPάᛶ䜢่⃭䛧䛯᮲௳䛷ヨ㦂䜢⾜䛳䛯䠊䜎䛯䠈⣽⬊

ᩘ䛾ῶᑡ䛻䜘䜚䝹䝅䝣䜵䝷䞊䝊άᛶ䛜పୗ䛩䜛ྍ⬟ᛶ䜢㝖እ䛩䜛┠ⓗ䛷䠈⣽⬊⏕Ꮡ⋡䜒䛒䜟䛫䛶ホ౯䛧䛯䠊

㑅ู䛥䜜䛯Ⰻዲ䛺άᛶ䜢♧䛩໬ྜ≀䛻䛴䛔䛶䛿䠈␲㝧ᛶ䜢㝖䛟┠ⓗ䛷䠈ኚ␗ᆺTCF⤖ྜ㡿ᇦ䠄CCTTTGGCC䠅䜢 䜒䛴 SuperFOPFlash

䜢୍㐣ᛶᑟධ䛧䛯⣽

⬊䜢⏝䛔䛯䝹䝅䝣䜵 䝷䞊䝊άᛶ䠄FOP ά ᛶ䠅䜢䛒䜟䛫䛶ホ౯ 䛧䛯䠊㝧ᛶヨᩱ䛷䛒 䜜䜀䠈FOPάᛶ䛻ᙳ 㡪䛩䜛䛣䛸䛺䛟 TOP ά ᛶ 䛾 䜏 䜢 㑅 ᢥ ⓗ 䛻ῶᑡ䛥䛫䜛䛸⪃䛘 䜙䜜䜛䠊䛧䛛䛧䠈ഇ㝧 ᛶヨᩱ䛾ሙྜ䠈䝹䝅 䝣䜵䝷䞊䝊䛯䜣䜁䛟 䛾Ⓨ⌧䜔ศゎ䛺䛹䛻

ᙳ㡪䛩䜛䛸⪃䛘䜙䜜䠈TOPάᛶ䛸䛸䜒䛻FOPάᛶ䜒పୗ䛩䜛䛣䛸䛜⪃䛘䜙䜜䜛䠊䛧䛯䛜䛳䛶FOPάᛶ䛚䜘䜃⣽⬊⏕Ꮡ

⋡䜢ኚ໬䛥䛫䜛䛣䛸䛺䛟䠈TOPάᛶ䛾䜏䜢ῶᑡ䛥䛫䜛䜒䛾䛜TCF/β-catenin㌿෗㜼ᐖ䠄Wnt䝅䜾䝘䝹㜼ᐖ䠅άᛶ䜢䜒䛴䜒 䛾䛸ุ᩿䛷䛝䜛䠄ᅗ2䠅䠊

ᡃ䚻䛿TOPάᛶ䜢ᣦᶆ䛸䛧䛯䝇䜽䝸䞊䝙䞁䜾䛻䛚䛔䛶㜼ᐖస⏝䠈䜎䛯䛿άᛶ໬స⏝䜢♧䛩ᩘ✀䛾᳜≀ᢳฟ≀䜢ぢ ฟ䛧䛯䠊ḟ䛻䛣䜜䜙䛾ᢳฟ≀䛛䜙ᚓ䛯άᛶᡂศ䛸䛭䜜䜙䛾Wnt䝅䜾䝘䝹㜼ᐖάᛶ䛻䛴䛔䛶⤂௓䛩䜛䠊

ᅗ2㻌Wnt䝅䜾䝘䝹⤒㊰

4.Calotropis gigantea䛛䛛䜙ᚓ䜙䜜䛯Wnt䝅䜾䝘䝹㜼ᐖᡂศ⁶

䜺䜺䜲䝰⛉᳜≀Calotropis gigantea䠄࿴ྡ䠖䜹䜲䜺䞁䝍䝞䝁䠅䛿ᖖ⥳పᮌ䛷䜰䝆䜰䠈䜰䝣䝸䜹䛾⇕ᖏ䠈ள⇕ᖏᇦ䛻ᗈ䛟 ศᕸ䛩䜛䠊ᮏ᳜≀䛾ⁱฟᾮ䝯䝍䝜䞊䝹ᢳฟ≀䛿䠈䝇䜽䝸䞊䝙䞁䜾ヨ㦂䛻䛚䛔䛶5 μg/mL䛾⃰ᗘ䛷TOPάᛶ䜢䜋䜌䝊䝻 䛻పୗ䛥䛫䛯䛜䠈⣽⬊⏕Ꮡ⋡䛾㢧ⴭ䛺పୗ䛿ㄆ䜑䜙䜜䛺䛛䛳䛯䠊௚䛾ᢳฟ≀䛻ẚ䜉䜘䜚ప⃰ᗘ䛷TOP㜼ᐖάᛶ䜢

♧䛧䛯䛣䛸䛛䜙䠈ᙉຊ䛺άᛶ໬ྜ≀䛾Ꮡᅾ䛜ᮇᚅ䛥䜜䠈䛭䛾ᡂศ᥈⣴䜢⾜䛳䛯䠊䝯䝍䝜䞊䝹ᢳฟ≀䜢⁐፹ศ㓄䛧䛶ᚓ 䜙䜜䛯㓑㓟䜶䝏䝹ྍ⁐㒊䛻άᛶ䛜ㄆ䜑䜙䜜䛯䛣䛸䛛䜙䠈άᛶヨ㦂䜢ᣦᶆ䛸䛧䛶䠈䝅䝸䜹䝀䝹䠈ODS䜹䝷䝮䛻䜘䜛䛥䜙䛺䜛 ศ⏬䜢㐍䜑䛯䠊䛭䛾⤖ᯝ䠈භ✀䛾䜹䝹䝕䝜䝷䜲䝗㢮 (1–6)䜢༢㞳䛧䠈᰾☢Ẽඹ㬆䠄NMR䠅䛚䜘䜃㉁㔞ศᯒ䠄MS䠅䛺䛹䛾 䝇䝨䜽䝖䝹䝕䞊䝍䛾ゎᯒ䛚䜘䜃ᩥ⊩್䛸䛾ẚ㍑䛻䜘䜚䛣䜜䜙䛾໬Ꮫᵓ㐀䜢Ỵᐃ䛧䛯䠄ᅗ3䠅䠊䛣䜜䜙䛿ᕥഃ䛾3’఩䛾⨨

᥮ᇶ䛜䛭䜜䛮䜜␗䛺䛳䛶䛔䛯䛜䠈10఩䛻䜰䝹䝕䝠䝗ᇶ䜢䜒䛴ඹ㏻䛩䜛㦵᱁䜢䜒䛴䠊

䛣䜜䜙໬ྜ≀䛿nM䜸䞊䝎䞊䛷TOPάᛶ䜢㜼ᐖ䛧䠈䛭䛾IC50್䛿0.7–^{3.8 nM䛷䛒䛳䛯䛜䠈FOPάᛶ䜢㢧ⴭ䛻ప}

ୗ䛥䛫䜛䛣䛸䛿䛺䛛䛳䛯䠊䜎䛯䠈1䠈3䠈5䠈6 䛿⣽⬊⏕Ꮡ⋡䜢పୗ䛥䛫䜛䛣䛸䛺䛟䠈TOP άᛶ䜢పୗ䛥䛫䛶䛔䛯䛣䛸䛛䜙䠈 TCF/β-catenin㌿෗㜼ᐖάᛶ䜢᭷䛧䛶䛔䜛䛸ุ᫂䛧䛯䠄ᅗ3䠅䠊୍᪉䠈2䛸4䛻䛴䛔䛶䛿TOPάᛶ䛾IC50್௜㏆䛾⃰ᗘ 䛻䛚䛔䛶ヨ㦂䛻⏝䛔䛯STF/293⣽⬊䛾⣽⬊⏕Ꮡ⋡䛾పୗ䛜ㄆ䜑䜙䜜䛯䛣䛸䛛䜙䠈TOPάᛶ䛾ῶᑡ䛿䠈TCF/β-catenin

㌿෗㜼ᐖ䛻㉳ᅉ䛩䜛䜒䛾䛷䛿䛺䛟䠈⣽⬊ᩘ䛾ῶᑡ䛻䜘䜛䜒䛾᥎ᐃ䛧䛯䠊

ḟ䛻⣽⬊ቑṪ䛜Wnt䝅䜾䝘䝹䛻౫Ꮡ䛧䛶䛔䜛኱⭠䛜䜣⣽⬊䛻ᑐ䛩䜛1–6䛾ᙳ㡪䜢ㄪ䜉䛯䠊䛣䛣䛷䛿䠈APC䛾Ḟኻ ኚ␗ᰴ䛷䛒䜛SW480䛚䜘䜃DLD1⣽⬊䛸䠈β-catenin䛻ኚ␗䜢䜒䛴HCT116⣽⬊䜢⏝䛔䛯䠊ẚ㍑ᑐ㇟䛸䛧䛶Wnt䝅䜾 䝘䝹㠀౫Ꮡᛶ䛾኱⭠䛜䜣RKO⣽⬊䜢⏝䛔䛯䠊Wnt䝅䜾䝘䝹䜢㜼ᐖ䛩䜛໬ྜ≀䛿Wnt㠀౫Ꮡᛶ⣽⬊䛻䛿ᙳ㡪䛩䜛 䛣䛸䛺䛟䠈Wnt౫Ꮡᛶ⣽⬊䛻ᑐ䛧䛶䛭䛾⣽⬊⏕Ꮡ⋡䜢㑅ᢥⓗ䛻పୗ䛥䛫䜛䛸⪃䛘䜙䜜䜛䠊

භ✀䛾໬ྜ≀䛾䛖䛱䠈஬✀ (1䠈3–^{6)䛿 1-10 nM 䛾⃰ᗘ䛻䛚䛔䛶 RKO}⣽⬊䛾⣽⬊⏕Ꮡ⋡䜢పୗ䛥䛫䜛䛣䛸䛺䛟䠈 Wnt౫Ꮡᛶ䛾SW480䠈DLD1䠈HCT116⣽⬊䛾⣽⬊⏕Ꮡ⋡䜢పୗ䛥䛫䛯䠄IC50್䠖1.8–7.0 nM䠅䠊䛣䛾⤖ᯝ䛛䜙䠈1䠈3䠈 5䠈6䛻䜘䜛Wnt䝅䜾䝘䝹౫Ꮡᛶ⣽⬊䛾⏕Ꮡ⋡䛾పୗ䛿䠈Wnt䝅䜾䝘䝹䛻㉳ᅉ䛩䜛䜒䛾䛸♧၀䛥䜜䛯䠊

ᅗ3㻌 ໬ྜ≀1-6䛾ᵓ㐀ᘧ䠈TCF/䃑-catenin㌿෗㜼ᐖస⏝

䛣䛾䛖䛱calotropin (1)䛾Wnt䝅䜾䝘䝹㜼ᐖస⏝䛾ศᏊᶵᵓ䛾ゎ᫂䜢┠ⓗ䛸䛧䛶Wnt䝅䜾䝘䝹㛵㐃䝍䞁䝟䜽䜈䛾ᙳ 㡪䜢᳨ウ䛧䛯䠊䛿䛨䜑䛻䠈Wnt䝅䜾䝘䝹䛾ㄪ⠇䛻㔜せ䛺㌿෗άᛶ໬ᅉᏊβ-catenin䜈䛾ᙳ㡪䜢䜴䜵䝇䝍䞁䝤䝻䝑䝖䛻䜘䜚 ㄪ䜉䛯䠊䛭䛾⤖ᯝ䠈SW480⣽⬊䛻䛚䛔䛶1 䛿⃰ᗘ౫Ꮡⓗ䛻β-catenin䜢ῶᑡ䛥䛫䛶䛚䜚䠈᰾ෆ䛚䜘䜃⣽⬊㉁䛻䛚䛔 䛶䜒ῶᑡ䛜ㄆ䜑䜙䜜䛯䠄ᅗ4A䠅䠊β-catenin䛿䠈๓㏙䛾䜘䛖䛻⣽⬊㉁ෆ䛻䛚䛔䛶2䛴䛾䜻䝘䞊䝊CK1α䠈GSK3β䛻䜘䛳 䛶䝸䞁㓟໬䛥䜜䠈䝥䝻䝔䜰䝋䞊䝮

⣔䛻䜘䜚ศゎ䛥䜜䜛䠊୍᪉䠈ศゎ 䛥䜜䛺䛔β-catenin䛿᰾ෆ䜈⛣⾜

䛧䠈TCF䛺䛹䛾ศᏊ䛸」ྜయ䜢ᙧ ᡂᚋ䠈DNA 䛻⤖ྜ䛧䠈Wnt 䝅䜾 䝘䝹䛾ᶆⓗ㑇ఏᏊ䛾㌿෗䜢ஹ 㐍䛩䜛䠊䜒䛧䠈TCF/β-catenin ㌿

෗㜼ᐖάᛶ䜢䜒䛴໬ྜ≀䛜䠈᰾

ෆβ-catenin䛾䜏䜢ῶᑡ䛥䛫䛯ሙྜ䠈A) β-catenin䛾᰾ෆ⵳✚䛾ᢚไ䛜Wnt䝅䜾䝘䝹㜼ᐖ䠄TCF/β-catenin㌿෗㜼ᐖ䠅 䛾స⏝ᶵᗎ䛸⪃䛘䜙䜜䜛䠊⣽⬊㉁䛚䜘䜃᰾䛾୧᪉䛷 β-catenin 䜢ῶᑡ䛥䛫䛺䛔ሙྜ䛿䠈B) ᰾ෆ䛻䛚䛔䛶TCF䛺䛹䛸 䛾」ྜయᙧᡂ䛾㜼ᐖ䠈䜒䛧䛟䛿ᶆⓗ㑇ఏᏊ䛾䝥䝻䝰䞊䝍䞊㡿ᇦ䛻䛚䛡䜛㌿෗ㄪ⠇㡿ᇦ䛸䛾⤖ྜ䛾㜼ᐖ䛜୺䛺స⏝

ᶵᗎ䛸⪃䛘䜙䜜䜛䠊䛧䛛䛧calotropin (1)䛿䠈᰾ෆ䛚䜘䜃⣽⬊㉁䛾཮᪉䛻䛚䛔䛶β-catenin䜢ῶᑡ䛥䛫䛯䛣䛸䛛䜙䠈A)䠈B) 䛾ᶵᵓ䛷䛿䛺䛟䠈C) ⣽⬊㉁ෆ䛻䛚䛔䛶β-catenin䜢ῶᑡ䛥䛫䠈䛭䛾⤖ᯝ䛸䛧䛶᰾ෆ⛣⾜䛩䜛β-catenin䛜ῶᑡ䛧Wnt 䝅䜾䝘䝹䠄TCF/β-catenin㌿෗䠅䛜㜼ᐖ䛥䜜䜛䜒䛾䛸⪃䛘䜙䜜䛯䠊1䛻䜘䜛Wnt䝅䜾䝘䝹䛾㜼ᐖ䛿䠈ᮏ䝅䜾䝘䝹䛾ᶆⓗ㑇 ఏᏊ䛷䛒䜛 c-myc 䝍䞁䝟䜽䝺䝧䝹䛜䠈1 䛾⃰ᗘ䛻౫Ꮡ䛧䛶పୗ䛧䛶䛔䛯䛣䛸䛛䜙䜒ᨭᣢ䛥䜜䛯䠊⣽⬊㉁ෆ䛷 β-catenin 䛜ῶᑡ䛩䜛せᅉ䛸䛧䛶䠈䝥䝻䝔䜰䝋䞊䝮䝅䝇䝔䝮䛻䛚䛡䜛 β-catenin 䛾ศゎಁ㐍䛜⪃䛘䜙䜜䜛䛣䛸䛛䜙䠈䝥䝻䝔䜰䝋䞊䝮 㜼ᐖ๣䛷䛒䜛MG-132䜢⏝䛔䛶᳨ウ䜢⾜䛳䛯䠊1䛾䜏䛾ῧຍ䛷䛿β-catenin䛿ῶᑡ䛧䛯䛜䠈1䛸MG-132䛸䜢ే⏝䛩䜛 䛸β-catenin䛾ῶᑡ䛿ㄆ䜑䜙䜜䛺䛛䛳䛯䛣䛸䛛䜙䠄ᅗ4B䠅䠈1䛻䜘䜚䝥䝻䝔䜰䝋䞊䝮⣔䛻䛚䛡䜛β-cateninศゎ䛜ಁ㐍䛥䜜 䛯䛸⪃䛘䜙䜜䛯䠊

APC➼䛸䛾β-cateninศゎ」ྜయෆ䛻䛚䛔䛶䠈䝥䝷䜲䝭䞁䜾䜻䝘䞊䝊CK1α䛻䜘䜚β-catenin䛾Ser45ṧᇶ䛜䝸䞁㓟

໬䛥䜜䜛䛸䠈GSK3β䛿β-catenin䛾Thr41/Ser37/Ser33ṧᇶ䜢㡰ḟ䝸䞁㓟໬䛩䜛䠊Ser37䠈Ser33䛾䝸䞁㓟໬䜢┠༳䛸䛧䛶 β-transducin repeat containing protein䠄β-TrCP䠅䛻䜘䜚β-catenin䛿䝫䝸䝴䝡䜻䝏䞁໬䛥䜜䠈䝥䝻䝔䜰䝋䞊䝮⣔䛷ศゎ䛥䜜 䜛²䠊

ᅗ4㻌1䛿䝥䝻䝔䜰䝋䞊䝮⣔䜢௓䛧䛶β-catenin䜢ศゎ䛩䜛

A) SW480⣽⬊䛻䛚䛔䛶1䛿⃰ᗘ౫Ꮡⓗ䛻β-catenin䛚䜘䜃c-myc䜢ῶᑡ䛥䛫䛯 B)1䛾β-cateninศゎ䛿䝥䝻䝔䜰䝋䞊䝮㜼ᐖ๣MG-132䛻䜘䜚ጉ䛢䜙䜜䛯

ᅗ5㻌 1ࡢSW480⣽⬊࡟࠾ࡅࡿβ-cateninࡢศゎ㸪ࣜࣥ㓟໬࡟ᑐࡍࡿᙳ㡪 A) LiClࢆ⏝࠸ࡓ᳨ウ B) CKI-7ࢆ⏝࠸ࡓ᳨ウ C) CK1α siRNAࢆ⏝࠸ࡓ᳨ウ

䛣䛾䛣䛸䜢䜅䜎䛘䠈ḟ䛻 1 䛾 β-catenin 䝸䞁㓟໬䜈䛾ᙳ㡪䛻䛴䛔䛶ゎᯒ䜢⾜䛳䛯䛸䛣䜝䠈1 䛿 β-catenin 䛾 Ser45䠈 Ser33/Ser37/Thr41䛾䝸䞁㓟໬䜢ಁ㐍䛧䠈䛭䛾⤖ᯝβ-catenin䜢ῶᑡ䛥䛫䛯䠊GSK3β㜼ᐖ๣䛷䛒䜛LiCl䛸1䛸䜢ే⏝䛩 䜛䛸䠈β-catenin䛾GSK3β䛻䜘䜛Ser33/Ser37/Thr41䝸䞁㓟໬䛿ᡴ䛱ᾘ䛥䜜䛯䛜䠈CK1α䛻䜘䜛Ser45䝸䞁㓟໬䛿ቑຍ䛧 䛯䜎䜎䛷䛒䜚䠈β-catenin䛾ῶᑡ䛿㉳䛣䜙䛺䛛䛳䛯䠄ᅗ

5A䠅䠊䜎䛯CK1α㜼ᐖ๣䛷䛒䜛CKI-7䛸1䛸䜢ే⏝䛩䜛䛸1䛻䜘䜚ಁ㐍䛥䜜䛯CK1α䛸GSK3β䛻䜘䜛䝸䞁㓟໬䛿ᢚไ䛥

䜜䠈β-catenin䛾ῶᑡ䜒㉳䛣䜙䛺䛛䛳䛯䠄ᅗ5B䠅䠊siRNA䛻䜘䜚CK1α䜢䝜䝑䜽䝎䜴䞁䛧䛯᮲௳䛷䛿1䜢ῧຍ䛧䛶䜒CK1α 䛸GSK3β䛻䜘䜛䝸䞁㓟໬䛿ㄆ䜑䜙䜜䛪䠈β-catenin䛾ῶᑡ䜒㉳䛣䜙䛺䛛䛳䛯䠄ᅗ5C䠅䠊䛥䜙䛻1䛿GSK3β䛾䝍䞁䝟䜽䝺䝧 䝹䛿ኚ໬䛥䛫䛪䠈CK1α䛾䝍䞁䝟䜽䝺䝧䝹䛚䜘䜃mRNA䜢ቑຍ䛥䛫䜛䛣䛸䛜ุ᫂䛧䛯䠊௨ୖ䛾䛣䛸䛛䜙1䛿CK1α䜢㌿

෗䝺䝧䝹䛷ቑຍ䛥䛫䜛䛣䛸䛻䜘䜚β-catenin䝸䞁㓟໬䜢ಁ㐍䛩䜛䠊䛣䜜䛻䜘䜚β-catenin䛾䝥䝻䝔䜰䝋䞊䝮⣔䛷䛾ศゎ䜢ಁ

㐍䛧䠈䛭䛾⤖ᯝ䠈Wnt䝅䜾䝘䝹䜢㜼ᐖ䛩䜛䛸♧၀䛥䜜䛯䠊

CK1α䛻స⏝䛩䜛పศᏊ䛿䠈㥑⹸⸆䛸䛧䛶⏝䛔䜙䜜䛶䛔䜛 pyrvinium⁷䜔䝣䜵䝙䝹䝥䝻䝟䝜䜲䝗䛷䛒䜛honokiol⁸䛜ሗ

࿌䛥䜜䛶䛔䜛䠊pyrvinium䛿CK1α䛾㓝⣲άᛶ䜢άᛶ໬䛧䠈honokiol䛿GSK3β䛸CK1α䛾୧᪉䜢䝍䞁䝟䜽䝺䝧䝹䛷ቑ ຍ䛥䛫䜛స⏝䜢䜒䛴䠊ᮏ◊✲䛷ぢฟ䛧䛯calotropin (1)䛿䠈pyrvinium䜔honokiol䛸␗䛺䜚GSK3β䛻䛿ᙳ㡪䜢୚䛘䜛䛣 䛸䛺䛟 CK1α 䜢䝍䞁䝟䜽䝺䝧䝹䛷ቑຍ䛥䛫䜛స⏝ᶵᵓ䜢䜒䛴䠊䛣䛾䜘䛖䛺స⏝䜢䜒䛴໬ྜ≀䛿䛣䜜䜎䛷䛻ሗ࿌䛜䛺䛟䠈

calotropin (1)䛿䝴䝙䞊䜽䛺స⏝ᶵᵓ䜢䜒䛴໬ྜ≀䛸⪃䛘䜙䜜䜛䠊

5.Xylocarpus granatum䛛䛛䜙ᚓ䜙䜜䛯Wnt䝅䜾䝘䝹㜼ᐖᡂศ⁹

䝉䞁䝎䞁⛉䛻ᒓ䛩䜛䝬䞁䜾䝻䞊䝤X. granatum䛿࿴ྡ䜢䝩䜴䜺䞁䝠䝹䜼䛸䛔䛔䠈䜰䝆䜰䠈䜰䝣䝸䜹䠈ኴᖹὒᆅᇦ䛻ᗈ䛟 ศᕸ䛩䜛䠊䝇䜽䝸䞊䝙䞁䜾ヨ㦂䛾⤖ᯝ䠈ᙉຊ䛺TCF/β-catenin㌿෗㜼ᐖάᛶ䠄TOPάᛶ㜼ᐖ䠅䜢♧䛧䛯ᮏ᳜≀ⴥ㒊䛾䝯 䝍䝜䞊䝹ᢳฟ≀䛻䛴䛔䛶άᛶ䜢ᣦᶆ䛻⁐፹ศ㓄䛺䜙䜃䛻ྛ✀䜹䝷䝮䜽䝻䝬䝖䜾䝷䝣䜱䞊䠄䝅䝸䜹䝀䝹䠈ODS䠈䝉䝣䜯䝕䝑

䜽䝇LH-20䛺䛹䠅䛻䜘䜚ศ㞳⢭〇䜢㐍䜑䛯䛸䛣䜝䠈䝦䜻䝃䞁ྍ⁐㒊䜘䜚ᅄ✀䛾䝸䝰䝜䜲䝗㢮 (7–10)䜢༢㞳䛧䛯䠊ྛ✀஧ḟ

ඖNMR䜔MS䝇䝨䜽䝖䝹䝕䞊䝍䛻ᇶ䛵䛟ᵓ㐀ゎᯒ䛾⤖ᯝ䠈໬ྜ≀7䠈8䛿᪂つ䛷䛒䜚䠈䛭䜜䛮䜜xylogranin A (7)䛚

䜘䜃B (8)䛸࿨ྡ䛧䛯䠊7䛿䝯䜻䝅䜹䝜䝷䜲䝗㦵᱁䠈 8䠈9䛿䝣䝷䜾䝬䝸䞁㦵᱁䛻ศ㢮䛥䜜䜛䠊䠄ᅗ6䠅

໬ྜ≀8䠈9䛾TOPάᛶ䛾IC50್䛿䛭䜜䛮䜜48.9䠈54.2 nM䛷䛒䛳䛯䠊୍᪉໬ྜ≀7䛿㜼ᐖάᛶ䜢♧䛥䛺䛛䛳䛯䠊

໬Ꮫᵓ㐀䛻╔┠䛩䜛䛸䠈໬ྜ≀8䠈9䛿8,9,30఩䛻䜸䝹䝖䜶䝇䝔䝹ᇶ䜢䜒䛴䛜䠈7䛻䛿䛺䛔䠊NOESY䛾ゎᯒ䛾⤖ᯝ䠈8 䛻䛚䛔䛶䛿5఩䛸17఩㛫䛻NOE┦㛵䛜ほ 䛥䜜䠈୧⪅䛿✵㛫ⓗ䛻㏆䛔఩⨨㛵ಀ䛻䛒䜛䛣䛸䛜♧၀䛥䜜䛯䠊䛥䜙䛻䠈

ᅗ6㻌໬ྜ≀7-10ࡢᵓ㐀ᘧ࡜DFTィ⟬

㸦B3LYP/6-31G*㸧࡟ࡼࡾᚓࡽࢀࡓ7㸪 8ࡢᏳᐃᵓ㐀

ᅗ6 ໬ྜ≀7-10ࡢᵓ㐀ᘧ࡜DF 㸦B3LYP/6-31G*㸧࡟ࡼࡾᚓࡽࢀ

8ࡢᏳᐃᵓ㐀

DFTィ⟬䜘䜚Ᏻᐃᵓ㐀䜢᳨ウ䛧䛯䛸䛣䜝䠈໬ྜ≀8 䛿䛛䛤≧䛾ᙧ≧䜢䛧䛶䛚䜚䠈H-5/H-17㛫䛾㊥㞳䛿2.8ύ䛸ẚ㍑ⓗ

㏆䛔㊥㞳䛻䛒䜛䛸ุ᫂䛧䠈7఩䛾䝯䝖䜻䝅ᇶ䛿17఩䛸ྠ䛨ഃ䠈䛩䛺䜟䛱concaveഃ䛻Ꮡᅾ䛩䜛䛣䛸䛜♧၀䛥䜜䛯䠊୍᪉䠈 7䛻䛚䛔䛶䛿H-5/H-17㛫䛾㊥㞳䛜6.9ύ䛷䛒䜛䛣䛸䠈䛚䜘䜃7఩䛾䝯䝖䜻䝅ᇶ䛜17఩䛸䛿㏫ഃ䠄convexഃ䠅䛻Ꮡᅾ䛩 䜛䛣䛸䛜♧၀䛥䜜䠄ᅗ6䠅䠈䛣䜜䜙䛾ᵓ㐀ⓗ䛺㐪䛔䛜7䠈8䛾άᛶ䛾㐪䛔䛻ᐤ୚䛧䛶䛔䜛䛸᥎ᐃ䛧䛯䠊

໬ྜ≀8, 9 䛻䛴䛔䛶኱⭠䛜䜣⣽⬊䠄SW480䠈HCT116䠈DLD1䠅䛻ᑐ䛩䜛⣽⬊ẘᛶ䜢᳨ウ䛧䛯䛸䛣䜝䠈䛣䜜䜙䛾໬ྜ

≀䛿ẚ㍑ᑐ㇟䛸䛧䛶䛒䜟䛫䛶ホ౯䛧䛯䝠䝖⫾ඣ⭈⣽⬊HEK293⣽⬊䛻ẚ䜉䠈Wnt౫Ꮡᛶ䛾SW480䛸HCT116⣽⬊

䛻ᑐ䛧䛶䜘䜚ᙉ䛔⣽⬊ẘᛶ䜢♧䛧䛯䠊

ḟ䛻䠈໬ྜ≀8䛾స⏝䜢᫂䜙䛛䛻䛩䜛┠ⓗ䛷䠈SW480⣽⬊䜢⏝䛔௨ୗ䛾ゎᯒ䜢⾜䛳䛯䠊β-catenin䛾䝍䞁䝟䜽䝺䝧 䝹䜢᳨ウ䛧䛯䛸䛣䜝䠈⣽⬊㉁䛷䛿㢧ⴭ䛺ኚ໬䛿♧䛥䛺䛛䛳䛯䛜䠈᰾ෆ䛻䛚䛔䛶䛿⃰ᗘ౫Ꮡⓗ䛻ῶᑡ䛧䛯䠄ᅗ 7A䠅䠊䜎 䛯䠈ච␿ᰁⰍἲ䛻䜘䜚β-catenin䛾ᒁᅾ䜢ほᐹ䛧䛯䛸䛣䜝䠈ᑐ↷⩌䛷䛿᰾ෆ䛻ㄆ䜑䜙䜜䛯β-catenin䛜䠈8䛾ῧຍ䛻䜘䜚

᰾䛛䜙ῶᑡ䛩䜛ഴྥ䛜ㄆ䜑䜙䜜䛯䠊௨ୖ䛾䛣䛸䛛䜙 8䛿䠈᰾䛾β-catenin 䛾⵳✚䜢ᢚไ䛩䜛స⏝䜢䜒䛴䛸⪃䛘䜙䜜䛯䠊 䛥䜙䛻Wnt䝅䜾䝘䝹䛾ᶆⓗ㑇ఏᏊ䛷䛒䜛c-myc䛚䜘䜃PPARδ䛻ᑐ䛩䜛ᙳ㡪䜢᳨ウ䛧䛯䛸䛣䜝䠈8䛿200 nM䛻䛚䛔 䛶⣽⬊඲య䛚䜘䜃᰾䛾c-myc䠈PPARδ䜢䝍䞁䝟䜽䝺䝧䝹䛷ῶᑡ䛥䛫䛯䠄ᅗ7B䠅䠊䜎䛯䠈8䛿ప⃰ᗘ䠄50-100 nM䠅䛻䛚䛔

䛶䛿c-myc䛾mRNA䜢ቑຍ䛥䛫䜛䜒䛾䛾䠈200 nM䛻䛚䛔䛶䛿䠈c-myc䠈PPARδ䛾mRNA䜢ῶᑡ䛥䛫䛯䠊௨ୖ䛾䛣䛸

䛛䜙8䛿Wnt䝅䜾䝘䝹䛾㌿෗άᛶ໬ᅉᏊ䛷䛒䜛β-catenin䛾᰾ෆ䛷䛾⵳✚䜢㜼ᐖ䛩䜛䛣䛸䛻䜘䜚Wnt䝅䜾䝘䝹䛾ᶆⓗ

㑇ఏᏊ䛾㌿෗䜢ᢚไ䛧䠈Wnt䝅䜾䝘䝹䜢㜼ᐖ䛩䜛䛣䛸䛜♧၀䛥䜜䛯䠊

䝸䝰䝜䜲䝗໬ྜ≀䛿⣽⬊ẘᛶ䜢䛿䛨䜑䠈᪻⹸䛻䛚䛡䜛ᦤ㣗㜼ᐖస⏝䛺䛹ᵝ䚻䛺⏕≀άᛶ䜢䜒䛴䛣䛸䛜ሗ࿌䛥䜜䛶䛔 䜛䛜¹⁰䠈Wnt䝅䜾䝘䝹䛾㜼ᐖస⏝䛻㛵䛩䜛ሗ࿌䛿௒ᅇ䛜ึ䜑䛶䛷䛒䛳䛯䠊

8䠈9䛾䜘䛖䛺䜸䝹䝖䜶䝇䝔䝹ᇶ䜢䜒䛴䝣䝷䜾䝬䝸䞁ᆺ䝸䝰䝜䜲䝗䛿X. granatum䛚䜘䜃䛭䛾㏆⦕✀䜘䜚ᩘ✀ሗ࿌䛥䜜䛶

䛚䜚^{10, 11}䠈䛭䛾άᛶᵓ㐀┦㛵䛻⯆࿡䛜ᣢ䛯䜜䜛䠊

6. Eurycoma longifolia䛛䛛䜙ᚓ䜙䜜䛯Wnt䝅䜾䝘䝹㜼ᐖᡂศ¹²

E. longifolia䛿䠈ᮾ༡䜰䝆䜰䛻ᗈ䛟ศᕸ䛩䜛䝙䜺䜻⛉䛾℺ᮌ䛷䠈䝬

䝺䞊䝅䜰䛷䛿䝖䞁䜹䝑䝖䜰䝸䠄Tongkat Ali䠅䛸࿧䜀䜜䠈ᙉኊ䜔ゎ⇕䛺䛹䜢

┠ⓗ䛻Ẹ㛫⸆䛸䛧䛶⏝䛔䜙䜜䛶䛔䜛䠊䝇䜽䝸䞊䝙䞁䜾䛻䛚䛔䛶䝠䝑䝖䛧 䛯ᮏ᳜≀䛾᰿㒊䝯䝍䝜䞊䝹ᢳฟ≀䛻䛴䛔䛶䠈άᛶ䜢ᣦᶆ䛸䛧䛶ศ⏬

䜢 㐍 䜑 䛯 䛸 䛣 䜝 䠈β 䜹 䝹 䝪 䝸 䞁 ᆺ 䜰 䝹 䜹 䝻 䜲 䝗 䛷 䛒 䜛 9-hydroxycanthin-6-one (11)䠈 䛚 䜘 䜃 䛭 䛾 㓄 ⢾ య 䛷 䛒 䜛 canthin-6-one 9-O-β-glucopyranoside (12)䠈䝸䝰䝜䜲䝗䛷䛒䜛 20,21,22,23-tetrahydro-23-oxoazadirone (13)䜢άᛶ໬ྜ

≀䛸䛧䛶༢㞳䛧䛯䠄ᅗ8䠅䠊11-13䛿䛭䜜䛮䜜6.8䠈11.6䠈3.0 μM䛷TCF/β-catenin㜼ᐖάᛶ䜢♧䛧䛯䠊䜎䛯䠈䛣䜜䜙໬ྜ

≀䛾ྛ✀⣽⬊䛻ᑐ䛩䜛⣽⬊ẘᛶ䜢ホ౯䛧䛯䛸䛣䜝䠈Wnt౫Ꮡᛶ䛾኱⭠䛜䜣⣽⬊䛻ᑐ䛧䛶ᙉ䛔⣽⬊ẘᛶ䜢♧䛧䛯䠊䛸䛟 䛻11䛚䜘䜃13䛿௚䛾Wnt౫Ꮡᛶ኱⭠䛜䜣⣽⬊䛻ẚ䜉䛶SW480⣽⬊䛻ᑐ䛧䛶䜘䜚ᙉ䛔ẘᛶ䜢♧䛧䛯䠊

ᅗ7 A)8ࡣSW480⣽⬊࡟࠾࠸࡚᰾ෆβ-cateninࢆῶᑡࡉࡏࡓ B)8ࡢWntࢩࢢࢼࣝᶆⓗࢱࣥࣃࢡ࡬ࡢᙳ㡪

ᅗ8㻌 ໬ྜ≀11-13䛾ᵓ㐀ᘧ

䛣䛾䛖䛱11䛻䛴䛔䛶䠈స⏝ᶵᗎ䛾ゎᯒ䜢㐍䜑䛯䠊β-catenin㔞䜈䛾ᙳ㡪䜢᳨ウ䛧䛯䛸䛣䜝䠈ᮏ໬ྜ≀䛿⣽⬊㉁䠈᰾䠈 ୧᪉䛻䛚䛔䛶 β-catenin 㔞䜢ῶᑡ䛥䛫䛯䠊ḟ䛻䝸䞁㓟໬䜈䛾ᙳ㡪䜢᳨ウ䛧䛯䛸䛣䜝䠈 11 䛿 GSK3β 䛻䜘䜛䝸䞁㓟໬

䠄p-β-catenin䠄S33/S37/T41䠅䠅䜢ቑຍ䛥䛫䛯䠊䛧䛛䛧 CK1α 䛻䜘䜛䝸䞁㓟໬䠄p-β-catenin 䠄S45䠅䠅䛿ኚ໬䛧䛶䛚䜙䛪䠈䛥䜙 䛻GSK3β䠈CK1α⮬య䛾䝍䞁䝟䜽㔞䜒ኚ໬䛧䛶䛔䛺䛛䛳䛯䠊䛣䛾⤖ᯝ䛛䜙䠈11䛿䠈GSK3β䛾㓝⣲άᛶ䜢ቑຍ䛥䛫䜛䛣䛸 䛜᥎ᐃ䛥䜜䛯䠄ᅗ9A䠅䠊䛣䛾௬ㄝ䛿௨ୗ䛻♧䛩GSK3β䛾siRNA䛻䜘䜛䝜䝑䜽䝎䜴䞁ᐇ㦂䛛䜙ᨭᣢ䛥䜜䛯䠊11䛿control siRNA䛾Ꮡᅾୗ䛷䛿β-catenin䜢ῶᑡ䛥䛫䛯䛜䠈GSK3β siRNAᏑᅾୗ䛷䛿11䛻䜘䜚ㄏᑟ䛥䜜䛯β-catenin䛾ῶᑡ䛿 ᡴ䛱ᾘ䛥䜜䛯䠊Ser 9ṧᇶ䛜䝸䞁㓟໬䛥䜜୙άᛶ໬䛥䜜䛯GSK3β䠄p-GSK3β䠄S9䠅䠅䝺䝧䝹䛻䛴䛔䛶ㄪ䜉䛯䛸䛣䜝䠈11䛿

control siRNA䛾Ꮡᅾୗ䛷䛿ῶᑡ䛧䛶䛔䛯䠊䛣䛾᮲௳䛷䛿GSK3β䛾䝍䞁䝟䜽䝺䝧䝹䛿ኚ໬䛧䛶䛔䛺䛛䛳䛯䛣䛸䛛䜙䠈11

䛿GSK3β䜢άᛶ໬䛧䠈β-catenin䛾䝸䞁㓟໬䜢ಁ㐍䛩䜛䛣䛸䛷β-catenin䛾ศゎ䜢ಁ㐍䛩䜛䜒䛾䛸♧၀䛥䜜䛯䠄ᅗ9B䠅䠊 ๓㏙䛾䜘䛖䛻β-catenin䛾ศゎ䛻䛿GSK3β䛻䜘䜛Ser33䠈Ser37䠈Thr41ṧᇶ䛾䝸䞁㓟໬䛻ඛ❧䛱䠈䝥䝷䜲䝭䞁䜾䜻䝘䞊

䝊CK1α䛻䜘䜛Ser45ṧᇶ䛾䝸䞁㓟໬䛜ᚲせ䛷䛒䜛䛸⪃䛘䜙䜜䛶䛔䜛䠊䛩䛺䜟䛱β-catenin䛾ศゎ䛿CK1α䛻౫Ꮡ䛧䛯

ᶵᵓ䛷䛒䜛䛸⪃䛘䜙䜜䛶䛔䜛䠊䛧䛛䛧䠈11 䛿 p-β-catenin䠄S45䠅䜢ቑຍ䛥䛫䜛䛣䛸䛺䛟䠈p-β-catenin䠄S33/S37/T41䠅䜢ቑຍ 䛥䛫䛶䛔䛯䠊䛧䛯䛜䛳䛶䠈CK1α䛻౫Ꮡ䛧䛺䛔β-catenin䛾ศゎᶵᵓ䛾Ꮡᅾ䛜⪃䛘䜙䜜䛯䠊䛭䛣䛷䠈CK1α䛾siRNA䜢

⏝䛔䛯䝜䝑䜽䝎䜴䞁ᐇ㦂䜢⾜䛳䛯䠊䛭䛾⤖ᯝ䠈control siRNAᏑᅾୗ䛻ㄆ䜑䜙䜜䛯11䛻䜘䜛p-β-catenin䠄S33/S37/T41䠅 䛾ቑຍ䛸䠈β-catenin䛾ῶᑡ䛿䠈CK1α siRNAᏑᅾୗ䛻䛚䛔䛶䜒ྠᵝ䛻ㄆ䜑䜙䜜䛯䠄ᅗ9C䠅䠊௨ୖ䛾䛣䛸䛛䜙11䛻䜘䜛 β-catenin䛾ศゎಁ㐍䛻䛿CK1α䛜㛵୚䛫䛪䠈GSK3β䛾άᛶ໬䜢௓䛧䛶Wnt䝅䜾䝘䝹䜢㜼ᐖ䛧䛶䛔䜛䛣䛸䛜♧၀䛥䜜 䛯䠊䛣䛾䜘䛖䛺స⏝䜢䜒䛴໬ྜ≀䛿⌧ᅾ䛾䛸䛣䜝ሗ࿌䛥䜜䛶䛚䜙䛪䠈䜎䛯䠈CK1α㠀౫Ꮡⓗ䛻β-catenin䜢ศゎ䛩䜛ᶵᵓ 䛻䛴䛔䛶䜒ሗ࿌䛜䛺䛔䛣䛸䛛䜙䠈ᮏ໬ྜ≀䛾స⏝䛿⯆࿡῝䛔䜒䛾䛸⪃䛘䜙䜜䜛䠊

ḟ䛻ᮏ໬ྜ≀䛾స⏝䛻䛴䛔䛶䝊䝤䝷䝣䜱䝑䝅䝳䜢⏝䛔䛯in vivoᐇ㦂䛻䛚䛔䛶᳨ウ䛧䛯䛸䛣䜝䠈ᮏ໬ྜ≀䛾ᢞ୚䛻䜘䜚䠈

୰⬻ᚋ⬻ቃ⏺䠄MHB䠖mid-hindbrain boundary䠅䛾ᙧᡂ୙඲䠈Ⰽ⣲⣽⬊䛾㢧ⴭ䛺ῶᑡ䜔䠈ᑿ㒊䛾ᒅ᭤䛺䛹䛾Wnt䝅䜾 䝘䝹㜼ᐖ䛸㛵㐃䛩䜛⾲⌧ᆺ䠄䝣䜵䝜䝍䜲䝥䠅䛾ኚ໬䛜ㄆ䜑䜙䜜䛯䠊䛥䜙䛻䝊䝤䝷䝣䜱䝑䝅䝳䛻䛚䛡䜛Wnt䝅䜾䝘䝹䛾ᶆⓗ

㑇ఏᏊ䛷䛒䜛zic2a䛚䜘䜃mitf䛻ᑐ䛩䜛ᙳ㡪䜢in situ hybridization䜢⏝䛔䛶ㄪ䜉䛯䛸䛣䜝䠈䛣䜜䜙ᶆⓗ㑇ఏᏊ䛾Ⓨ⌧

䛾పୗ䛜☜ㄆ䛷䛝䛯䠊zic2a 䠄zinc finger protein of the cerebellum2a䠅䛿୰⬻䛾Ⓨ㐩䛻㛵୚䛧䛶䛚䜚䠈୰⬻ど⵹䛺䛹 䛷Ⓨ⌧䛜ㄆ䜑䜙䜜䜛䠊䜎䛯䠈mitf䠄microphthalmia-associated transcription factor䠅䛿⚄⤒ሐ⣽⬊䛻Ⓨ⌧䛜ㄆ䜑䜙䜜䠈⚄

⤒ሐ⣽⬊䛛䜙Ⰽ⣲⣽⬊䜈䛾ศ໬ㄏᑟ䜢ไᚚ䛩䜛䛣䛸䛜▱䜙䜜䛶䛔䜛䠊䛣䜜䜙ᶆⓗ㑇ఏᏊ䛾పୗ䛜๓㏙䛾䝣䜵䝜䝍䜲䝥 䛾ኚ໬䜈ᐤ୚䛧䛶䛔䜛䛣䛸䛜♧၀䛥䜜䛯䠊๓㏙䛧䛯㏻䜚䠈11䛿in vitro ᐇ㦂䛻䛚䛔䛶GSK3β䛾άᛶ໬䜢௓䛧䛶Wnt 䝅䜾䝘䝹䜢㜼ᐖ䛧䛶䛔䜛䛣䛸䛜♧၀䛥䜜䛯䠊䛭䛣䛷䝊䝤䝷䝣䜱䝑䝅䝳䛻䛚䛡䜛11䛾Wnt㜼ᐖస⏝䛜GSK3β䛻㛵୚䛧䛶

ᅗ9㻌 11ࡢSW480⣽⬊࡟࠾ࡅࡿβ-cateninࡢศゎ㸪ࣜࣥ㓟໬➼࡟ᑐࡍࡿᙳ㡪 A) ࣜࣥ㓟໬࡟ᑐࡍࡿ᳨ウ B) GSK3βࢆ⏝࠸ࡓ᳨ウ C) CK1α siRNAࢆ⏝࠸ࡓ᳨ウ

䛔䜛䛛䜢᳨ウ䛩䜛┠ⓗ䛷䠈GSK3β 㜼ᐖ๣䛷 䛒䜛 BIO䠄6-bromo-indirubin-3'-oxime䠅䜢⏝

䛔᳨ウ䜢⾜䛳䛯䠊Wnt 䝅䜾䝘䝹䛾άᛶ໬䛻 䜘䜚ㄆ䜑䜙䜜䜛䝊䝤䝷䝣䜱䝑䝅䝳䛾䝣䜵䝜䝍䜲䝥 䛾ኚ໬䛸䛧䛶┠䛾ᙧᡂ୙඲䛜▱䜙䜜䛶䛔䜛䠊 BIO༢⊂ฎ⌮䛷䛿GSK3β䛜㜼ᐖ䛥䜜䜛䛣䛸 䛻䜘䜚Wnt䝅䜾䝘䝹䛜άᛶ໬䛥䜜䜛䛯䜑䠈┠

䛾ᙧᡂ୙඲䛜☜ㄆ䛷䛝䜛䠊䛧䛛䛧䠈BIO䛸11䛸䜢ే⏝䛩䜛䛸䠈┠䛾ᙧᡂ䛜ᅇ᚟䛧䛯䝣䜵䝜䝍䜲䝥䛜☜ㄆ䛷䛝䠈in vivo䛻 䛚䛔䛶䜒ᮏ໬ྜ≀䛿GSK3β䜢௓䛧䛶Wnt䝅䜾䝘䝹䜢㜼ᐖ䛧䛶䛔䜛䛣䛸䛜♧၀䛥䜜䛯䠄ᅗ10䠅䠊䛣䜜䜙䛾⤖ᯝ䜢⥲ྜ䛩 䜛䛸䠈ᮏ໬ྜ≀䛿in vivo䠈in vitro཮᪉䛻䛚䛔䛶GSK3β䛾άᛶ໬䜢௓䛧䛶Wnt䝅䜾䝘䝹䜢㜼ᐖ䛧䛶䛔䜛䛸⪃䛘䜙䜜䛯䠊

7.Azadirachta excelsa䛛䛛䜙ᚓ䜙䜜䛯Wnt䝅䜾䝘䝹㜼ᐖస⏝䜢䜒䛴໬ྜ≀¹³

A. excelsa 䠄䝉䞁䝎䞁⛉䠅䛾ᯝᐇ䝯䝍䝜䞊䝹ᢳฟ≀䛿䠈10 μg/mL䛻䛚䛔䛶TOPάᛶ䜢⣙80%㜼ᐖ䛧䛯䠊TOP άᛶ䜢ᣦᶆ䛸䛧䛶䠈⁐፹ศ㓄䠈䝅䝸䜹䝀䝹䜹䝷䝮䠈㏫┦HPLC䛺䛹䛻䜘䜚ᮏᢳฟ≀䛾ศ⏬䜢㐍䜑䛯䠊䛭䛾⤖ᯝ䠈ά ᛶᡂศ䛸䛧䛶᪂つ໬ྜ≀ 14

䜢 ྵ䜐 ඵ✀䛾 ໬ྜ ≀ (14– 21) 䜢ᚓ䛯䠄ᅗ 11䠅䠊᰾☢Ẽ ඹ㬆䠄NMR䠅䠈㉁㔞ศᯒ䠄MS䠅 䛺䛹䛾ྛ✀䝇䝨䜽䝖䝹䝕䞊䝍 䛾ゎᯒ䛻ᇶ䛵䛝䠈䛣䜜䜙໬ྜ

≀䛾ᵓ㐀Ỵᐃ䠈ྠᐃ䜢⾜䛳䛯䠊 䛣䜜䜙໬ྜ≀䛾 TOP 㜼ᐖస

⏝䛿䠈7,14-oxetane, 15-䝠䝗䝻 䜻䝅య䛷䛒䜛14 (IC50 = 33.7 μM), 15 (20 μM)䛚䜘䜃䠈15-

䜿䝖䠈7-䝠䝗䝻䜻䝅య䛷䛒䜛20(14.8 μM), 21(36.7 μM)䛷䛿୰⛬ᗘ䛷䛒䛳䛯䛜䠈14,15-epoxy, 7-䝠䝗䝻䜻䝅య䛷䛒䜛 16–19䛷䛿nM䜸䞊䝎䞊䛸ᙉຊ䛷䛒䛳䛯 (16; 127 nM, 17; 300 nM,18; 252 nM,19; 121 nM)䠊

䛣䛾䛖䛱䠈17 䛻䛴䛔䛶⣽⬊ቑṪ䛜Wnt 䝅䜾䝘䝹౫Ꮡᛶ䛷䛒䜛䛜䜣⣽⬊䛻ᑐ䛩䜛⣽⬊ẘᛶ䜢᳨ウ䛧䛯䛸䛣䜝䠈

AGS䠄䝠䝖⫶䛜䜣䠅䠈HCT116䠄䝠䝖኱⭠䛜䜣䠅⣽⬊䛻ᑐ䛩䜛 IC50್䛿䛭䜜䛮䜜 239 nM䠈161 nM 䛷䛒䛳䛯䠊୍᪉

Wnt䝅䜾䝘䝹౫Ꮡᛶ䛷䛒䜛䝠䝖኱⭠䛜䜣 SW480䛚䜘䜃DLD1䠈䛚䜘䜃Wnt㠀౫Ꮡᛶ䛷䛒䜛䝠䝖኱⭠䛜䜣 RKO

⣽⬊䛻ᑐ䛩䜛IC50್䛿400 nM௨ୖ䛷䛒䛳䛯䠊䜴䜵䝇 䝍䞁䝤䝻䝑䝖䛻䜘䜚䛥䜙䛺䜛ゎᯒ䜢㐍䜑䛯⤖ᯝ䠈17 䛿 Wnt䝅䜾䝘䝹䛾㌿෗άᛶ໬ᅉᏊβ-catenin䛾⣽⬊㉁

䛚䜘䜃᰾䛷䛾䝍䞁䝟䜽䝺䝧䝹䜢ῶᑡ䛥䛫䜛䛣䛸䛺䛟䠈 Wnt 䝅䜾䝘䝹䛾ᶆⓗ䝍䞁䝟䜽䛷䛒䜛 c-myc 䛾᰾ෆ䝺 䝧䝹䜢ῶᑡ䛥䛫䜛䛣䛸䛷䠈Wnt 䝅䜾䝘䝹䜢㜼ᐖ䛩䜛䛸

♧၀䛥䜜䛯䠄ᅗ12䠅䠊 ᅗ11㻌 ໬ྜ≀14–^{21䛾ᵓ㐀ᘧ}

ᅗ12 18䛾β-catenin䠈c-myc䛻ᑐ䛩䜛ᙳ㡪㸦HCT116⣽⬊㸧

ᅗ10㻌ࢮࣈࣛࣇ࢕ࢵࢩࣗ࡟࠾ࡅࡿBIOㄏⓎ║ᙧᡂ୙඲࡟ᑐࡍࡿ11ࡢᙳ㡪

8.Ricinus communis䛛䛛䜙ᚓ䜙䜜䛯Wnt䝅䜾䝘䝹άᛶ໬స⏝䜢䜒䛴໬ྜ≀¹⁴ ᮏ䝇䜽䝸䞊䝙䞁䜾◊✲䛷䛿䠈Wnt㜼ᐖ䜢♧䛩໬ྜ≀䛰䛡䛷䛿䛺䛟䠈άᛶ

໬䛩䜛໬ྜ≀䜒ぢฟ䛧䛯䠊R. communis 䠄䝖䜴䝎䜲䜾䝃⛉䠅䛾ⱼ㒊䝯䝍䝜䞊 䝹ᢳฟ≀䛿䠈10 μg/mL䛻䛚䛔䛶⣙80%TOPάᛶ䜢ୖ᪼䛥䛫䜛స⏝䜢♧

䛧䛯䠊TOP άᛶ䜢ᣦᶆ䛸䛧䛶ศ⏬䜢㐍䜑䛯⤖ᯝ䠈άᛶᡂศ䛸䛧䛶 ricinin (22), 7-oxo-β-sitosterol (23) 䜢ྵ䜐ᅄ✀䛾ᡂศ䜢ศ㞳䛧䛯䠄ᅗ13䠅䠊22䛚 䜘䜃23䛿䜰䝑䝉䜲⣽⬊䛾⣽⬊⏕Ꮡ⋡䜢㢧ⴭ䛻ቑຍ䛥䛫䜛䛣䛸䛺䛟TOPά

ᛶ䜢ୖ᪼䛥䛫䛯䠊䜎䛯䠈䛣䜜䜙໬ྜ≀䛿FOPάᛶ䜢ኚ໬䛥䛫䛺䛛䛳䛯䛣䛸䛛䜙TCF/β-catenin㌿෗άᛶ໬స⏝䜢 䜒䛴䛣䛸䛜♧၀䛥䜜䛯䠊

䛣䛾䛖䛱22䛾స⏝䛻䛴䛔䛶䛥䜙䛻

᳨ウ䜢㐍䜑䛯⤖ᯝ䠈22 䛿 STF/293

⣽⬊䛻䛚䛔䛶⣽⬊඲య䠈⣽⬊㉁䠈

᰾䛻䛚䛔䛶 β-catenin 䛾䝍䞁䝟䜽㔞 䜢ቑຍ䛥䛫䛯䠊ḟ䛻β-catenin䛾䝸䞁 㓟໬䝺䝧䝹䜢ㄪ䜉䛯⤖ᯝ䠈CK1α 䛚䜘䜃GSK3β䛻䜘䜛β-catenin䛾䝸 䞁㓟໬䛿㢧ⴭ䛻ῶᑡ䛧䛶䛔䛯䜒䛾 䛾䠈䛣䜜䜙䜻䝘䞊䝊䛾䝍䞁䝟䜽䝺䝧

䝹䛾ኚ໬䛿ㄆ䜑䜙䜜䛺䛛䛳䛯䠄ᅗ14A䠅䠊䛣䛾⤖ᯝ䛛䜙22䛜CK1α䛾㓝⣲άᛶ䜢㜼ᐖ䛧䛶䛔䜛ྍ⬟ᛶ䛜⪃䛘䜙䜜䛯 䛯䜑䠈CK1α䛾άᛶ໬స⏝䜢䜒䛴pyrvinium䜢⏝䛔᳨ウ䛧䛯䠊䛭䛾⤖ᯝ䠈22䛸pyrvinium䛸䛾ే⏝䛻䜘䜚䠈pyrvinium ༢⊂ฎ⌮䛻䜘䜚ᘬ䛝㉳䛣䛥䜜䛯β-catenin䛾ῶᑡ䛸CK1α䛻䜘䜛β-catenin䛾䝸䞁㓟໬䛾ቑຍ䛜ᡴ䛱ᾘ䛥䜜䛯䛣䛸䛛䜙

䠄ᅗ14B䠅䠈22䛿CK1α䛾㓝⣲άᛶ䜢㜼ᐖ䛧䛶䛔䜛䛣䛸䛜♧၀䛥䜜䛯䠊䛥䜙䛻䝊䝤䝷䝣䜱䝑䝅䝳⬇䜢⏝䛔䛯in vivo䛷䛾

స⏝䜢᳨ウ䛧䛯䛸䛣䜝䠈22䛿β-catenin䜢ቑຍ䛥䛫䜛䛣䛸䛜ุ᫂䛧䛯䠊 㻌9. 䛚䜟䜚䛻

௨ୖ䠈ᡃ䚻䛾◊✲䛷ぢฟ䛧䛯 Wnt 䝅䜾䝘䝹䛻స⏝䛩䜛⏕≀άᛶኳ↛≀䛸䛭䜜䜙䛾άᛶⓎ⌧ᶵᵓ䛻䛴䛔䛶⤂௓䛧 䛯䠊ᚓ䜙䜜䛯୍㒊䛾άᛶኳ↛≀䛻䛴䛔䛶䛿䛭䛾స⏝ᶵᗎ䛾୍➃䜢᫂䜙䛛䛻䛩䜛䛣䛸䛜䛷䛝䛯䠊䛭䛾స⏝ᶵᗎ䜢

β-catenin䛻ᑐ䛩䜛ᙳ㡪䛻↔Ⅼ䜢ᙜ䛶䛶䜎䛸䜑䜛䛸௨ୗ䛾䜘䛖䛻䛺䜛䠊

1) β-catenin䛾ศゎ䛻స⏝䛩䜛䠖 1 䛿CK1α 䜢㌿෗䝺䝧䝹䛷ቑຍ䛥䛫䠈β-catenin 䛾䝸䞁㓟໬䠈ศゎ䜢ಁ㐍䛧䠈Wnt 䝅䜾䝘䝹䜢㜼ᐖ䛩䜛䠊11 䛿GSK3β䛾άᛶ໬䛻䜘䜚β-catenin 䛾䝸䞁㓟໬䠈ศゎ䜢ಁ㐍䛧䠈Wnt䝅䜾䝘䝹䜢㜼ᐖ䛩䜛䠊 22䛿 CK1α䛾㓝⣲άᛶ䜢㜼ᐖ䛩䜛䛣䛸䛻䜘䜚䠈β-catenin䛾䝸䞁㓟໬䠈ศゎ䜢ᢚไ䛧䠈Wnt䝅䜾䝘䝹䜢άᛶ໬䛩䜛䠊

2) β-catenin䛾᰾ෆ⵳✚䛻స⏝䛩䜛䠖 8䛿β-catenin䛾᰾ෆ⵳✚䜢ᢚไ䛩䜛䠊

3) β-catenin䛾䝍䞁䝟䜽䝺䝧䝹䛻䛿ᙳ㡪䛧䛺䛔䠖 17䛿β-catenin䝺䝧䝹䜢ኚ໬䛥䛫䛺䛛䛳䛯䛜䠈Wnt䝅䜾䝘䝹䛾ᶆⓗ

㑇ఏᏊ䛾Ⓨ⌧䜢పୗ䛥䛫䜛䛸♧၀䛥䜜䛯䠊

䛣䛾䜘䛖䛻䠈ᮏ◊✲䛷䛿 β-catenin 䛻ᑐ䛧䛶ᵝ䚻䛺స⏝Ⅼ䛷ᙳ㡪䛧Wnt䝅䜾䝘䝹䛻స⏝䛩䜛⏕≀άᛶኳ↛≀䜢ぢ ฟ䛧䛯䠊Wnt 䝅䜾䝘䝹䛻ᑐ䛧䛶స⏝䛩䜛་⸆ရ䛿䛔䜎䛰ୖᕷ䛥䜜䛯䜒䛾䛿䛺䛟䠈⌧ᅾ䛔䛟䛴䛛䛾⸆≀䛾⮫ᗋヨ㦂䛜

⾜䜟䜜䛶䛔䜛䠊䜎䛯䠈Wnt䝅䜾䝘䝹⤒㊰䛾ไᚚᶵᵓ䛿䛔䜎䛰୙᫂䛺㒊ศ䜒ከ䛔䛸䛥䜜䛶䛔䜛䠊Wnt䝅䜾䝘䝹䛻స⏝䛩 ᅗ14 22䛾STF/293⣽⬊䛻䛚䛡䜛β-cateninࡢศゎ㸪ࣜࣥ㓟໬➼࡟ᑐࡍࡿᙳ㡪 A) ࣜࣥ㓟໬࡟ᑐࡍࡿ᳨ウ B) pyrviniumࢆ⏝࠸ࡓ᳨ウ

ᅗ13㻌 ໬ྜ≀22䠈23䛾ᵓ㐀ᘧ

䜛పศᏊ໬ྜ≀䛾᥈⣴䛿⢭ຊⓗ䛻⾜䜟䜜䛶䛔䜛䜒䛾䛾䛭䛾ከ䛟䛜ྜᡂ໬ྜ≀䜢ᑐ㇟䛸䛧䛯䜒䛾䛷䛒䜚䠈ኳ↛≀䜢ᑐ

㇟䛸䛧䛯 Wnt 䝅䜾䝘䝹䛻స⏝䛩䜛ኳ↛≀䛾᥈⣴౛䛿ከ䛟䛺䛔䠊௒ᅇ⤂௓䛧䛯໬ྜ≀䜔䛭䛾㛵㐃໬ྜ≀䜢ά⏝䛧䛶 Wnt䝅䜾䝘䝹䛾ᮍゎ᫂䛺㒊ศ䜢᫂䜙䛛䛩䜛䛣䛸䛻䜘䜚䠈⏕࿨⛉Ꮫ◊✲䜔་⸆ရ㛤Ⓨ䜈㈉⊩䛧䛯䛔䠊

㻌 ㅰㅰ㎡䠖ᮏ◊✲䛿༓ⴥ኱Ꮫ኱Ꮫ㝔⸆Ꮫ◊✲㝔 άᛶᵓ㐀໬Ꮫ◊✲ᐊ䛷䛒䜛䠊䛤ᣦᑟ䛔䛯䛰䛝䜎䛧䛯▼ᶫṇᕫᩍᤵ䠈Ⲩ஭⥳

෸ᩍᤵ䛻῝䛟ឤㅰ⏦䛧ୖ䛢䜎䛩䛸䛸䜒䛻䠈◊✲ᐊ䝯䞁䝞䞊䛾ດຊ䛻῝䛟ឤㅰ䛔䛯䛧䜎䛩䠊ᮏ◊✲䜢㐙⾜䛩䜛䛻䛒䛯䜚䠈᳜≀

᥇ྲྀ䛻䛤༠ຊ䛟䛰䛥䛔䜎䛧䛯䜽䝹䝘኱Ꮫ䠄䝞䞁䜾䝷䝕䝅䝳䠅Samir K. Sadhuᩍᤵ䠈䝎䝑䜹኱Ꮫ䠄䝞䞁䜾䝷䝕䝅䝳䠅Firoj Ahmedᩍ

ᤵ䠈䝔䝮䝁䞉䝁䞊䝫䝺䞊䝅䝵䞁ᑠ㇂㔝ႛ༤ኈ䠈䝁䞁䜿䞁኱Ꮫ䠄䝍䜲䠅Thaworn Kowithayakornᩍᤵ䠈STF/293⣽⬊䜢䛤ᜨ୚䛟䛰 䛥䛔䜎䛧䛯䝆䝵䞁䝩䝥䜻䞁䝇኱Ꮫ Jeremy Nathans ᩍᤵ䠈SuperFOP-Flash 䝥䝷䝇䝭䝗䜢䛤ᜨ୚䛟䛰䛥䛔䜎䛧䛯䝽䝅䞁䝖䞁኱Ꮫ

Randall Moon ᩍᤵ䛻ᚰ䛛䜙ឤㅰ⏦䛧ୖ䛢䜎䛩䠊䝊䝤䝷䝣䜱䝑䝅䝳䛾ᐇ㦂䛻㝿䛧䜎䛧䛶ከ኱䛺䛤ᣦᑟ䠈䛤ᨭ᥼䜢䛔䛯䛰䛝䜎䛧

䛯༓ⴥ኱Ꮫ኱Ꮫ㝔⸆Ꮫ◊✲㝔 ⏕໬Ꮫ◊✲ᐊ䛾ఀ⸨⣲⾜ᩍᤵ䠈⁁ཱྀ㈗ṇ༤ኈ䛻῝ㅰ䛔䛯䛧䜎䛩䠊ᮏ◊✲㐙⾜䛻ᙜ䛯䜚䛤 ᨭ᥼䜢㈷䜚䜎䛧䛯᪥ᮏᏛ⾡᣺⯆఍⛉Ꮫ◊✲㈝䠈䝁䝇䝯䝖䝻䝆䞊◊✲᣺⯆㈈ᅋ䠈℈ཱྀ⏕໬Ꮫ᣺⯆㈈ᅋ䠈Ṋ⏣⛉Ꮫ᣺⯆㈈ᅋ䛻 ឤㅰ䛔䛯䛧䜎䛩䠊

ཧ⪃ᩥ⊩

1. Ueda, M., Chemical biology of natural products on the basis of identification of target proteins. Chem. Lett. 2012,41, 658-666.

2. Anastas, J. N.; Moon, R. T., WNT signalling pathways as therapeutic targets in cancer. Nat. Rev. Cancer 2013,13, 11-26.

3. ⳥ụ ❶, Wntࢩࢢࢼࣝࢿࢵࢺ࣮࣡ࢡ࡜ࡑࡢ␗ᖖ࡟ࡼࡿ⑓ែ. ⏕໬Ꮵ 2009,81, 780-792.

4.Clevers, H.; Nusse, R., Wnt/β-catenin signaling and disease. Cell 2012,149, 1192-1205.

5. Li, X.; Ohtsuki, T.; Koyano, T.; Kowithayakorn, T.; Ishibashi, M., New Wnt/β-catenin signaling inhibitors isolated from Eleutherine palmifolia.Chem. Asian J. 2009,4, 540-547.

6. Park, H. Y.; Toume, K.; Arai, M. A.; Sadhu, S. K.; Ahmed, F.; Ishibashi, M., Calotropin: A Cardenolide from Calotropis giganteathat Inhibits Wnt Signaling by Increasing Casein Kinase 1α in Colon Cancer Cells. ChemBioChem 2014,15, 872-878.

7. Thorne, C. A.; Hanson, A. J.; Schneider, J.; Tahinci, E.; Orton, D.; Cselenyi, C. S.; Jernigan, K. K.; Meyers, K. C.; Hang, B.

I.; Waterson, A. G.; Kim, K.; Melancon, B.; Ghidu, V. P.; Sulikowski, G. A.; LaFleur, B.; Salic, A.; Lee, L. A.; Miller, D.

M.; Lee, E., Small-molecule inhibition of Wnt signaling through activation of casein kinase 1α. Nat. Chem. Biol. 2010,6, 829-836.

8. Singh, T.; Katiyar, S. K., Honokiol inhibits non-small cell lung cancer cell migration by targeting PGE2-mediated activation ofβ-catenin signaling. PLoS One 2013,8, e60749.

9. Toume, K.; Kamiya, K.; Arai, M. A.; Mori, N.; Sadhu, S. K.; Ahmed, F.; Ishibashi, M., Xylogranin B: A Potent Wnt Signal Inhibitory Limonoid from Xylocarpus granatum.Org. Lett. 2013,15, 6106-6109.

10. Tan, Q.-G.; Luo, X.-D., Meliaceous Limonoids: Chemistry and Biological Activities. Chem. Rev. 2011,111, 7437-7522.

11. Liao, S.-G.; Chen, H.-D.; Yue, J.-M., Plant Orthoesters. Chem. Rev. 2009,109, 1092-1140.

12. Ohishi, K.; Toume, K.; Arai, M. A.; Koyano, T.; Kowithayakorn, T.; Mizoguchi, T.; Itoh, M.; Ishibashi, M.,

9-Hydroxycanthin-6-one, a β-Carboline Alkaloid from Eurycoma longifolia, Is the First Wnt Signal Inhibitor through Activation of Glycogen Synthase Kinase 3β without Depending on Casein Kinase 1α. J. Nat. Prod.2015,78, 1139-1146.

13. Fuentes, R. G.; Toume, K.; Arai, M. A.; Sadhu, S. K.; Ahmed, F.; Ishibashi, M., Limonoids with Wnt signal inhibitory activity isolated from the fruits of Azadirachta excelsa.Phytochem. Lett. 2015,11, 280-285.

14. Ohishi, K.; Toume, K.; Arai, M. A.; Sadhu, S. K.; Ahmed, F.; Mizoguchi, T.; Itoh, M.; Ishibashi, M., Ricinine: A pyridone alkaloid from Ricinus communisthat activates the Wnt signaling pathway through casein kinase 1α. Bioorg. Med. Chem.

2014,22, 4597-4601.

各部門・附属センターの活動と業績

⏕

⏕⸆㈨※⛉Ꮫศ㔝 Division of Pharmacognosy

㻌

ᩍ ᤵ ᑠᯇ࠿ࡘᏊ Professor Katsuko Komatsu (Ph.D.)

෸ᩍᤵ ␜ 㖭 ୍ ᩥ Associate Professor Kazufumi Toume (Ph.D.)

ຓ ᩍ ᮒ ጨ Assistant Professor Shu Zhu (Ph.D.)

◊✲ဨ ⴱ ㌍ ೧ Postdoctoral Fellow Yuewei Ge (Ph.D.)

ᢏ⬟⿵బဨ ᖾ 㞞 Ꮚ Assistant Masako Yuki

஦ົ⿵బဨ ᪂ ஭ ᜨ Ꮚ Assistant Keiko Arai 㻌

ۍ

ۍ◊◊✲┠ⓗ 㻌

ᆅ⌫⎔ቃࡢኚ໬࡟ࡼࡾ㸪⸆⏝ኳ↛㈨※ࡢῶᑡࡀ༴᝹ࡉࢀࡿࠋࡑࡇ࡛ᮏศ㔝࡛ࡣ㸪⏕⸆㈨※ࡢ

⌧≧ࡢᢕᥱ࡜௦᭰⏕⸆ࡢ㛤Ⓨ㸪⏕⸆ࡢ≉ᚩࢆᢕᥱࡋࡓຠ⋡ⓗ฼⏝ࡢಁ㐍୪ࡧ࡟᱂ᇵ⸆⏝᳜≀ࡢ 㑅ᢥ࡜᱂ᇵᣑ඘ࢆ┠ⓗ࡟ࡋ࡚㸪࢔ࢪ࢔࡟࠾ࡅࡿ₎⸆㈨※ࡢㄪᰝ࡜⸆⏝⏕≀ࡢ㑇ఏᏛⓗ㸪ᡂศ໬

Ꮫⓗ㸪⸆⌮Ꮫⓗከᵝᛶࡢゎᯒࢆ⾜࠺ࠋࡲࡓ㸪ኳ↛⸆≀ࡢᶆ‽໬ࢆ┠ⓗ࡟ࡋ࡚㸪㑇ఏᏊከᆺ࡟ᇶ

࡙ࡃ⏕⸆ྠᐃἲࡢ㛤Ⓨ୪ࡧ࡟ᡂศ࣭άᛶ᝟ሗࡢ⼥ྜ࡟ࡼࡿ⏕⸆ᶵ⬟ࡢゎᯒࢆ⾜࠺ࠋ

ۍ

ۍ◊◊✲ᴫせ 㻌 Ϩ

Ϩ㸧⸆⸆⏝⏕≀ཬࡧఏ⤫⸆≀ࡢㄪᰝ◊✲㻌㻌

㻌 ࢖ࢠࣜࢫ㸪࢜ࣛࣥࢲ㸪ࢻ࢖ࢶ㸪ࣇࣛࣥࢫཬࡧࢫ࢙࣮࢘ࢹࣥࡢ᳜≀ᅬࢆゼၥࡋ㸪୰ᅜ࠿ࡽ࣮ࣚࣟ

ࢵࣃ࡟ࡶࡓࡽࡉࢀࡓRheumᒓ᳜≀ࡢㄪᰝࢆ⾜ࡗࡓࠋ ϩ

ϩ㸧⸆⸆⏝᳜≀࣭⏕⸆ࡢከᵝᛶࡢゎᯒ㻌㻌

㻌 1) Gentianaᒓ᳜≀13✀2ኚ✀ࡢ᰾rDNAࡢITS㡿ᇦࡢ㑇ఏᏊከᆺ࠿ࡽศ㢮ୖ᭷⏝࡞࣐࣮࣮࢝

㓄ิࢆぢฟࡋ㸪ᕷሙὶ㏻⏕⸆ࡢࠕ❳⫹ࠖཬࡧࠕ⛙Ⱄࠖࢆྠᐃࡋࡓࠋࡲࡓ㸪G. scabra⏤᮶ࡢ

❳⫹ཬࡧG. crassicaulis⏤᮶ࡢ⛙Ⱄ࠿ࡽ㸪᪂つ࡞ࢭࢥ࢖ࣜࢻ࢖ࢻ㓄⢾య12ᡂศࢆ༢㞳㸪ྠᐃ

ࡋࡓࠋࡑࢀࡽ࡟᪤▱21ᡂศࢆຍ࠼࡚㸪ᢠ⅖⑕άᛶࢆㄪ࡭ࡓࠋ

2) ㉥Ⱏࡢᢠ࢔ࣞࣝࢠ࣮ᡂศࡢ᥈⣴◊✲࡟ࡼࡾ㸪ࣔࣀࢸࣝ࣌ࣀ࢖ࢻ16ᡂศࢆ༢㞳㸪ྠᐃࡋࡓࠋ Ϫ

Ϫ㸧⸆⸆⏝᳜≀ࡢᅜෆ᱂ᇵࡢᣑᣑ඘ࢆࢆᣦྥࡋࡋࡓ◊✲㻌㻌

㻌㻌 ¹⁾ ࢩࣕࢡࣖࢡࡢ᰿ࡢㄪ〇ຍᕤἲ࡜ࡋ࡚㸪᥀ࡾୖࡆᚋࡢప ಖᏑ࡜‮㏻ࡋࡢ㔜せᛶࢆ࣌࢜ࢽࣇ

ࣟࣜࣥ㸪࣌ࣥࢱ࢞ࣟ࢖ࣝࢢࣝࢥ࣮ࢫ࡞࡝ࡢྵ㔞࠿ࡽ᫂ࡽ࠿࡟ࡋࡓࠋࡲࡓ㸪࢖࣓࣮ࢪࣥࢢ㉁

㔞ศᯒ࡟ࡼࡾ㸪࢔ࣝࣅࣇࣟࣜࣥ࡞࡝ࡢ஧ḟ௦ㅰ⏘≀ࡢ⤌⧊ෆศᕸࢆ᫂ࡽ࠿࡟ࡋࡓࠋ

2) ࢚ࢰ࢘ࢥࢠࡢ㣴ᾮ᱂ᇵࢆ⾜࠸㸪཰✭ࡉࢀࡓⴥ࡟࢝ࣇ࢙࢜࢖ࣝ࢟ࢼ㓟㢮࡞࡝10ᡂศࢆぢฟࡋ ࡓࠋ

ϫ

ϫ㸧₎᪉᪉๣࣭⏕⸆࣭೺ᗣ㣗ရࡢရ㉁࡜ࣞࢠ࣮ࣗࣞࢩࣙࣥ

㻌㻌 (⊂)་⸆ᇶ┙◊✲ᡤ⸆⏝᳜≀㈨※◊✲ࢭࣥࢱ࣮࡟༠ຊࡋ࡚㸪᪥ᮏ࡛ὶ㏻ࡋ࡚࠸ࡿ࢟ࣙ࢘࢝ࢶ㸪

ཬࡧࢣ࢖࢞࢖ࡢITS㡿ᇦࡢሷᇶ㓄ิࡢゎᯒࢆ⾜࠸㸪ࡑࢀࡽࡢᇶ※✀ࢆ᫂ࡽ࠿࡟ࡋࡓࠋ 㻌㻌

ۍ

ۍཎཎⴭㄽᩥ 㻌

㻌 ¹⁾ He J. Y., Zhu S., Komatsu K., Goda Y., Cai S. Q.: Genetic polymorphism of medicinally-used Codonopsisspecies in an internal transcribed spacer sequence of nuclear ribosomal DNA and its application to authenticate Codonopsis Radix. J. Nat. Med., 68: 112-24, 2014.

㻌 ²⁾ Wiriyakarun S., Zhu S., Komatsu K., Sukrong S.: The use of cycleave PCR for the differentiation

㈨

㈨

※

※ 㛤 㛤

Ⓨ

Ⓨ 㒊 㒊 㛛 㛛

資源開発部門

of the rejuvenating herb species Pueraria candollei(White Kwao Khruea), Butea superba(Red Kwao Khruea), and Mucuna macrocarpa(Black Kwao Khruea), and the simultaneous detection of multiple DNA targets in a DNA admixture. Nat. Prod. Commun., 9: 111-7, 2014.

㻌 ³⁾ He J. Y., Zhu S., Goda Y., Cai S. Q., Komatsu K.: Quality Evaluation of Medicinally-used Codonopsis Species and Codonopsis Radix Based on the Contents of Pyrrolidine Alkaloids, Phenylpropanoids and Polyacetylenes. J. Nat. Med., 68: 326-39, 2014.

㻌 ⁴⁾ Shimada K., Natamura Y., Kawase M., Komatsu K., Saito T., Takahashi K.: Establishment of a methodology for identifying Paeoniae Radix based on metallomics analysis. J. Nat. Med., 68:

407-13, 2014.

㻌 ⁵⁾ He J. Y., Zhu S., Komatsu K.: HPLC/UV Analysis of Polyacetylenes, Phenylpropanoid and Pyrrolidine Alkaloids in Medicinally Used Codonopsis Species. Phytochem. Anal., 25: 213-9, 2014.

㻌 ⁶⁾ Tsukahara K., Toume K., Ito H., Ishikawa N., Ishibashi M.:Isolation of β-indomycinone guided by cytotoxicity tests fromStreptomycessp. IFM11607 and revision of its double bond geometry. Nat.

Prod. Commun., 9: 1327-8, 2014.

㻌 ⁷⁾ Andoh T., Kitamura R., Fushimi H., Komatsu K., Shibahara N., Kuraishi Y.: Effects of goshajinkigan, hachimijiogan, and rokumigan on mechanical allodynia induced by paclitaxel in mice. J. Tradit. Complement. Med., 4: 293-7, 2014.

㻌 ۍ

ۍⴭ᭩㻌 㻌

㻌 ¹⁾ ᑠᯇ࠿ࡘᏊ㸪ᮒጨ㸸⏕⸆ࡢከᵝᛶゎᯒ࡜ᶆ‽໬㸬ᕝཎಙኵ┘ಟࠕ⸆⏝᳜≀࣭⏕⸆ࡢ᭱๓

⥺̿ᅜෆ᱂ᇵᢏ⾡࠿ࡽရ㉁ホ౯㸪〇ရ㛤Ⓨࡲ࡛ࠖ㸪ࢩ࣮࢚࣒ࢩ࣮㸪ᮾி㸪2014㸪p. 157-65.

㻌 ۍ

ۍ⥲⥲ㄝ 㻌

㻌 ¹⁾ Kuboyama T., Tohda C., Komatsu K.: Effects of Ashwagandha (roots of Withania somnifera) on neurodegenerative diseases. Current Topics, Biol. Pharm. Bull., 37: 892-7, 2014.

㻌 ²⁾ ␜㖭୍ᩥ㸸࣓ࣛࢽࣥ⏕ᡂ࡟࠿࠿ࢃࡿ࢘࢕ࣥࢺࢩࢢࢼࣝࢆㄪ⠇ࡍࡿኳ↛≀ࡢ᥈⣴㸬ࣇࣞࢢ

ࣛࣥࢫࢪ࣮ࣕࢼࣝ㸪2014㸪42: 61-70㸬

㻌 ³⁾ ␜㖭୍ᩥ㸪▼ᶫṇᕫ㸸࢘࢕ࣥࢺࢩࢢࢼࣝㄪ⠇ࢆ┠ᣦࡋࡓ⏕≀άᛶኳ↛≀ࡢ᥈⣴㸬໬Ꮫ࡜

⏕≀㸪2014㸪52: 731-41㸬 㻌

ۍ

ۍᏛᏛ఍ሗ࿌㻌㻌㸦㸦㻖㻖㻦㻌≉≉ูㅮ₇㸪ࢩ࣏ࣥࢪ࣒࢘㸪࣮࣡ࢡࢩࣙࢵࣉ➼㸧

㻖㻌 ¹⁾ Komatsu K., Zhu S., He J. Y.: Genetic and chemical polymorphism of medicinally-used Codonopsisspecies and its application to evaluate Codonopsis Radix. International Symposium on Herbal Medicine and Natural Resources, 2014, 2, 22-23, Kanazawa, Japan. (Invited lecture) 㻌 ²⁾ Zhu S., Shirakawa A., Shi Y. H., Yu X. L., Tamura T., Yoshimatsu K., Komatsu K.: Comparing

the contents of main components in the roots of Bonten, a medicinal cultivar of Paeonia lactiflora after different post-harvest processing. The 8th JSP-CCTCNM-KSP Joint Symposium on Pharmacognosy, 2014, 9, 13, Fukuoka, Japan.

㻌 ³⁾ Fushimi H., Fushimi N., Zhu S., Komatsu K.: A list book recording the precious articles in the Museum of Material Medica, 2014, 9, 13, Fukuoka, Japan.

㻌 4) Nakamura K., Ryuse M., Zhu S., Komatsu K., Hattori M., Iwashima M.: Analysis of DNA fragments encoding puerarin metabolizing enzyme from a human intestinal bacterium. The 8th JSP-CCTCNM-KSP Joint Symposium on Pharmacognosy, 2014, 9, 13, Fukuoka, Japan.

㻌 ⁵⁾ Ge Y. W., Kazuma K., Zhu S., Yoshimatsu K., Komatsu K.: Comprehensive Analysis of Sequencing Proanthocyanidin Oligomers in Rhubarb by HPLC-ESI-MSⁿ. The 8th JSP-CCTCNM-KSP Joint Symposium on Pharmacognosy, 2014, 9, 13, Fukuoka, Japan.

㻌 6) He Y. M., Zhu S., Wu X. T., Ge Y. W., Komatsu K.: Genetic and chemical diversity of Gentiana

plants and drugs (4) −Isolation of constituents with anti-inflammatory activity from Gentianae Macrophyllae Radix. The 8th JSP-CCTCNM-KSP Joint Symposium on Pharmacognosy, 2014, 9, 13, Fukuoka, Japan.

㻌 7) Harada K., Oda T., Sumi A., Ueno N., Kojoma M., Yamamoto Y., Zhu S., Komatsu K., Hirata K.:

Application of metabolomics for quality evaluation of licorice (Glycyrrhiza uralensis Fisher).

The 8th JSP-CCTCNM-KSP Joint Symposium on Pharmacognosy, 2014, 9, 13, Fukuoka, Japan.

㻌 ⁸⁾ Fushimi H., Fushimi N., Komatsu K.: Investigation of Japanese folk medicines for enriching the contents of the Ethnomedicine Database. The 8th JSP-CCTCNM-KSP Joint Symposium on Pharmacognosy, 2014, 9, 13, Fukuoka, Japan.

㻌 ⁹⁾ Zhu S., Yu X. L., Komatsu K.: Genetic and chemical charaterization of white and red peony root derived from Paeonia lactiflora. The 28th International Symposium on the Chemistry of Natural Products and the 8th International Conference on Biodiversity (ISCNP28 & ICOB8), 2014, 10, 19-24, Shanghai, China.

㻖㻌 ¹⁰⁾ Komatsu K.: Development of branded herbal drugs on the basis of analytical results on genetic and chemical diversity of medicinal plants. The 14th International Symposium on Traditional Medicine in Toyama (2014), 2014, 10, 27-28, Toyama, Japan.

㻖㻌 ¹¹⁾ Ishibashi M., Arai M. A., Toume K.: New heterocyclic natural products isolated in screening studies on disease and development pathways. The 3rd International Symposium on Chemical Biology of Natural Products: Target ID and Regulation of Bioactivity, 2014 10, 29, Osaka.

㻌 ¹²⁾ Fuentes R. G., Toume K., Arai M. A., Ishibashi M.: Scopadulciol, isolated from Scoparia dulcis, overcomes TRAIL resistance in AGS human gastric adenocarcinoma cells. The Annual Meeting of the Philippine Society for Biochemistry and Molecular Biology, 2014, 12, 4-5, Cebu City, Philippine.

㻖㻌 ¹³⁾ Toume K.: Search for bioactive natural products that affect on WNT signaling pathway. 2014 Joint Symposium of the Natural Products Research Institute at Seoul National University and the Institute of Natural Medicine at University of Toyama, 2014, 12, 10, Seoul, Korea.

㻌 ¹⁴⁾ ఱẙᩄ㸪ᮒጨ㸪࿋ᬡ፱㸪ᑠᯇ࠿ࡘᏊ㸸Gentianaᒓ⏕⸆ࡢᇶཎ࡜ရ㉁࡟㛵ࡍࡿ◊✲㸦1㸧

̿New secoiridoids from Gentianae Scabrae Radix.᪥ᮏ⸆Ꮫ఍➨134ᖺ఍㸪2014㸪3㸪27-30㸪

⇃ᮏ㸬

㻌 ¹⁵⁾ ࿋ᬡ፱㸪ᮒጨ㸪னᬡ㯇㸪ྜ⏣ᖾᗈ㸪ᑠᯇ࠿ࡘᏊ㸸Gentiana ᒓ⏕⸆ࡢᇶཎ࡜ရ㉁࡟㛵ࡍ

ࡿ◊✲㸦2㸧 ̿Gentianaᒓ4✀ཬࡧ❳⫹ࡢITS㓄ิ࡟ࡘ࠸࡚㸬᪥ᮏ⸆Ꮫ఍➨134ᖺ఍㸪 2014, 3, 27-30㸪⇃ᮏ㸬

㻌 ¹⁶⁾ ୰ᮧ㈼୍㸪❳℩┿⾰㸪῿⨾ᚿಖ㸪ᑠᯇ࠿ࡘᏊ㸪᭹㒊ᚁ㞝㸪ᒾᓥㄔ㸸ࣉ࢚ࣛࣜࣥ௦ㅰ཯

ᛂ࡟࠾ࡅࡿ㓝⣲཯ᛂ୰㛫యࡢྠᐃ㸬᪥ᮏ⸆Ꮫ఍➨134ᖺ఍㸪2014, 3, 27-30㸪⇃ᮏ㸬 㻌 17㸧 ῝㇂ᬕᙪ㸪㔜᳃※ኴ㸪୍ᰡᖾ⏕㸪ᮒጨ㸪ᑠᯇ࠿ࡘᏊ㸪➉㇂Ꮥ୍㸸Stemona-amine C, D ࡢ

⤯ᑐᵓ㐀࡟ࡘ࠸࡚㸬᪥ᮏ⸆Ꮫ఍➨134ᖺ఍㸪2014, 3, 27-30㸪⇃ᮏ㸬

㻌 18㸧 ⴱ㌍೧㸪ᮧୖᏲ୍㸪⏣ᮧ㝯ᖾ㸪ᕝᮏඖ⿱㸪☾⏣㐍㸪ᮒጨ㸪ྜྷᯇ჆௦㸪ᑠᯇ࠿ࡘᏊ㸸Chemical constituents analysis of the leaf of Eleutherococcus senticosus cultivated in different environment.➨31ᅇ࿴₎་⸆Ꮫ఍Ꮫ⾡኱఍㸪2014, 8, 31㸪༓ⴥ㸬

㻌 19㸧 ὸ἟⯙㸪୰すṇ೔㸪ᒣ㊰ㄔ୍㸪అぢ⿱฼㸪ᑠᯇ࠿ࡘᏊ㸸ᮏⲡᩥ⊩࡟ࡳࡽࢀࡿࠗ⣸ཧ࠘

ࡢᛶ≧グ㍕࡟ࡘ࠸࡚㸬➨31ᅇ࿴₎་⸆Ꮫ఍Ꮫ⾡኱఍㸪2014, 8, 31㸪༓ⴥ㸬

㻌 20㸧 ࿋ᬡ፱㸪ᮒጨ㸪ྜ⏣ᖾᗈ㸪ᑠᯇ࠿ࡘᏊ㸸Gentianaᒓ⏕⸆ࡢᇶཎ࡜ရ㉁࡟㛵ࡍࡿ◊✲㸦3㸧

̿Gentiana ᒓ8✀ཬࡧ⛙Ⱄࡢ ITS㓄ิ࡟ࡘ࠸࡚㸬᪥ᮏ⏕⸆Ꮫ఍➨ 61ᅇᖺ఍㸪2014, 9, 13-14㸪⚟ᒸ㸬

㻌 21㸧 ῝㇂ᬕᙪ㸪୍ᰡᖾ⏕㸪ᮒጨ㸪ᑠᯇ࠿ࡘᏊ㸸ࣅࣕࢡࣈࢥࣥࡼࡾᚓࡽࢀࡓ᪂つstemoninoamide ᆺ࢔ࣝ࢝ࣟ࢖ࢻࡢ⤯ᑐᵓ㐀࡟ࡘ࠸࡚㸬᪥ᮏ⏕⸆Ꮫ఍➨61ᅇᖺ఍㸪2014, 9, 13-14㸪⚟ᒸ㸬 㻌 22㸧 㛵㇂ዲ⮳㸪୍ᰡᖾ⏕㸪㔜᳃※ኴ㸪ἅ⃝ᑦᏊ㸪ᮒጨ㸪ᑠᯇ࠿ࡘᏊ㸪➉㇂Ꮥ୍㸸Stemona tuberosa

ࡼࡾᚓࡽࢀࡓ4⣭࢔ࣝ࢝ࣟ࢖ࢻࡢᵓ㐀㸬᪥ᮏ⏕⸆Ꮫ఍➨61ᅇᖺ఍㸪2014, 9, 13-14㸪⚟

ᒸ㸬

㻌 23㸧 ୰ᮧ㈼୍㸪ᒾᓥㄔ㸪❳℩┿⾰㸪⚟ཎ㞙㸪ᮒጨ㸪ᑠᯇ࠿ࡘᏊ㸪᭹㒊ᚁ㞝㸸ࣉ࢚ࣛࣜࣥC-㓄

⢾య௦ㅰ཯ᛂ࡟࠾ࡅࡿ㓝⣲཯ᛂ୰㛫యࡢᵓ㐀Ỵᐃ㸬᪥ᮏ⏕⸆Ꮫ఍➨61ᅇᖺ఍㸪2014, 9, 13-14㸪⚟ᒸ㸬