本土杉科與柏科植物成分及其活性之研究(2/2)

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行政院國家科學委員會專題研究計畫成果報告

本土杉科與柏科植物成分及其活性之研究(2/2)

研究成果報告(完整版)

計畫類別：個別型計畫編號： NSC 95-2113-M-002-004- 執行期間： 95 年 08 月 01 日至 96 年 07 月 31 日執行單位：國立臺灣大學化學系暨研究所計畫主持人：郭悅雄計畫參與人員：博士班研究生-兼任助理：林煥祐、邱錫臨碩士班研究生-兼任助理：林士傑、林明園、林文偉博士後研究：陳政琪報告附件：出席國際會議研究心得報告及發表論文處理方式：本計畫可公開查詢

中華民國 96 年 08 月 21 日

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1

行政院國家科學委員會補助專題研究計畫

行政院國家科學委員會補助專題研究計畫成果

成果

成果報告

成果

報告

※※※※※※※※※※※※※※※※※※※※※※※※※

※ 本土杉科與柏科植物成分及其活性研究 ※

※ (Studies on the Biological Components from Endemic ※

※ Plants of Taxodiaceae and Cupressaceae) ※

※※※※※※※※※※※※※※※※※※※※※※※※※

計畫類別：■個別型計畫 □整合型計畫

計畫編號：NSC －95－2113－M－002－004

執行期間：95 年 8 月 1 日至 96 年 7 月 31 日

計畫主持人：郭悅雄

共同主持人：

計畫參與人員：陳政琪，林煥祐，邱錫臨，林士傑，林明園，林文偉

成果報告類型(依經費核定清單規定繳交)：■精簡報告 □完整報告

本成果報告包括以下應繳交之附件：

□

赴國外出差或研習心得報告一份

□赴大陸地區出差或研習心得報告一份

■

出席國際學術會議心得報告及發表之論文各一份

□國際合作研究計畫國外研究報告書一份

執行單位：國立台灣大學化學系

中華民國 96 年 6 月 30 日

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行政院國家科學委員會專題研究計畫成果報告

本土杉科與柏科植物成分及其活性成分研究

Studies on the biological Components from Endemic

Plants of Taxodiaceae and Cupressaceae

計畫編號：NSC 94-2113-M-002-022

執行期限：95 年 8 月 1 日至 96 年 7 月 31 日

主持人：郭悅雄國立台灣大學化學系

計畫參與人員：陳政琪，林煥祐，邱錫臨，林士傑，林明

園，林文偉

一一一一、、、中文摘要、中文摘要中文摘要中文摘要台灣肖楠是台灣產針葉樹一級木之一種，也是特有變種，而且也是台灣重要的建築材料，由其皮部得到的化合物中有 4 個新的二帖雙聚化合物，以及二個新骨架的 C35 化合物，由扁柏分出一個木酚素 yatein 具有抗第一型泡疹病毒之功能。由薏仁之麩皮甲醇萃取物可以調降大鼠 granulosa cell 的黃體素。由台灣雲杉找出三種不同類型新化合物。三種水溶性新化合物及 4 種已知化合物由正榕之氣根分出，合成 salvinal 的新方法被研發出。苦瓜是保健食物發現其果實發現能活化 PPARα 之成份。另一種保健食物固體發酵樟芝首次發現 5 種新的 labdane type 之二帖擁有腦神經保護功能。另外發現牛樟子實體材之甲醇萃取物可以抑制肝癌細胞 Hep 3B 之成長。關鍵詞關鍵詞關鍵詞關鍵詞：台灣肖楠；薏仁; 台灣雲杉；正榕;苦瓜；牛樟；台灣扁柏；salvinol 合成 Abstract AbstractAbstract Abstract

Catocedrus marcolepis var. formosana, a member of the Cupressaceae, is an endemic plant and also an important building material in Taiwan. Studies the bark of this plant, four new dimeric compounds, calocedimers A (1) , B (2), C (3), and D (4）were found. The four

new compounds are dimers of

abietane-O-camphane, abietane-O-cadinane,

abietane-O-abietane, and abietane-O-abietane respectively. Same extract, two novel skeletonal compounds C35, were also isolated.

A lignan, yatein, isolated from

Chamaecyparis obtusa var. formosana

significantly suppressed HSV-1

multiplication in HeLa cell. Methanolic

extract of adlay bran exhibited

downregulation of progesterone biosythesis in rat granulose cells. Study on the heartwood of Picea morrisonicola found three new different skeleton compounds (monoterpene, aromatic and lignan). Three new water-soluble together with three known compounds were purified from the aerial roots of Ficus microcarpa. Salvinal is a novel adenosine A1 receptor ligand and

anti-cancer drug. We used a concise and efficient synthesis method for synthesis salvinal from isoeugenol. Momordica charantia L. is a functional food, we found an activator of PPARα from the bitter gourd. From another functional food, Antrodia

cinnamomea, five labdane derivatives ( including three new compounds) were isolated. The in vitr oneuroprectives activity of all compounds was evaluated. The methanolic extract from the fruiting bodies of

A. cinnamomea decreased cell proliferation of Hep 3B. From the heartwood of

Chamaecyparis obtusa var. formosana, four new diterpenes and three known diterpenes were isolated and elucidated.

Keywords: Calocedrus macrolepis var.

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3

obtusa var. formosana ; Picea

morrisonicola; Ficus microcarpa

Antradia camphorata

二二二

二、、、Purpose of Project、Purpose of ProjectPurpose of ProjectPurpose of Project

Yatein, a lignan, was isolated from C.

obtuse var. formosana, it inhibited herpes simplex virus type 1 (HSV-1) replication. This plant is an endemic conifer and also an important material in architecture, furniture and sculpture. Study on the component of this plant is very important for understanding this plant’s biosynthesis. Antradia camphorata, a parasitic fungus, possessed several biological activities such as immunomodulating, antioxidative, and hepatoprotective effect. It is necessary to studing the pure compounds, and identified which compound possesses what biological activity. Picer morrisonicola is a good building material, previous studies

discovered many novel compounds.

Therefore, further studies on different fraction attracted me to investigat again the new components. Twenty-four species of

Ficus are used in Chinese Herbal medicine F.

microcarpa is used for the treatment of rhenmatism-arthralgia, diarrhea and acariasis. These biological activities attracted me to study the chemical constituentd. Salvinal is a novel adenosine A1 receptor ligand and

strong anti-cancer agent. Three papers have described the method of synthesis. In this time, we used a concise and efficient synthesis of it from isoeugenol via phenoxenium ion intermediate. Bitter gourd has been used in traditional medicine for the treatment of diabetes. PPARs can control lipid and glucose homeostasis. It led me to study its components.

三三三

三、、、Content of Project、Content of ProjectContent of Project Content of Project

The separative procedure of every plant was achieved as following

plant

methanol methanol extract

partitioned with H₂O / EtOAc

EtOAc soluble fraction H2O

Yatein (1), a component of C. obtusa var.

formosana, can suppress herpes simplex

virus type 1 replication in Hela cells by interruption the immediate-early gene expression. In the study, the effects and the mechanisms of mathanolic extract of Adlay (Coix lachrymal-jobi L. var. ma-yuen stupf) bran on progesterone synthesis in rat granulusa cell were standied. And it was found the this extract did downregulation of progesterone biosynthesis. Two novel C35

terpenoids ferrugicadinol(2), ferrugieudesmol (3) and four calcedimers A (4), B (5), C (6), D (7) together with platydiol (8),α-cadinol (9), ferrugiol (10), and 6,7-dehydroferrugiol (11), were isolated from the bark of C. macrolepis var. formosana. Compounds 2 and 3 showed cytotoxicity against human oral epidermoid carcinoma KB cells with IC50 value ranging from 11 to 14µM. Three

new compounds, 19-

hydroxylabda-8(17)-en-16,15-olide (12), 3β,19-dihydroxylabda-8(17),11E-dien-16,15-olide (13), and 13-epi-3β,19-dihydroxylabda- 8(17),11E-dien-16,15-olide (14), together with four known compounds, 19-

hydroxylabda-8(17),13-dien-16,15-olide (15), 14-deoxy-11,12-didehydroandrographolide (16), 14-deoxyandrographolide, and

pinusolidic acid, were isolated from the fruiting bodies of Antrodia camphorata. The structures of compounds 1-3 were elucidated by the analysis of their spectroscopic data. The in vitro neuroprotective activity of all compounds was evaluated, and compounds

1-5 protected neurons from Aβdamage by

39.2, 35.0, 36.7, 30.6, and 27.0%,

respectively, at concentrations between 5 and 20µM. The purpose of this study was to evaluate the apoptotic effects of ethylacetate extract from A. cinnamomea (EAC) fruiting

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4

bodies in Hep 3B, a liver cancer cell line. EAC decreased cell proliferation of Hep 3B cells by inducing apoptotic cell death. EAC treatment increased the level of calcium (Ca2+) in the cytoplasm and triggered the subsequent activation of calpain and caspase-12. EAC also initiated the

mitochondrial apoptotic pathway through regulation of Bcl-2 family proteins expression, release of cytochrome c, and activation of caspase-9 in Hep 3B cells. Furthermore, the mitochondrial apoptotic pathway amplified the calpain pathway by Bid and Bax interaction and Ca2+ translocation. Three new constituents, including a p-menthane type monoterpene,

trans-p-menthane-7,8,9-triol (17), an

aromatic, 2,6-dihydroxy-3,4-dimethylbenzoic acid methyl ester (18), and a lignan,

(+)-morrisonicolanin (19), were isolated from the low polar layer of heartwood extracts of Picea morrisonicola, and their structures were elucidated on the basis of spectroscopic analysis. Of these compounds identified, 1 was obtained as its diacetylated derivative

Three new water-soluble constituents [ficuscarpanoside B (20), (7E,9Z)- dihydrophaseic acid 3-O-β-D-

glucopyranoside (21) and ficuscarpanic acid (22)] and the natural product 2,2’-dihydroxyl ether have been isolated, together with three known compounds [(7S,8R)-

syringoylglycerol, (7S,8R)-

syringoylglycerol-7-O-β-D-glucopyranoside and icariside D2] from the aerial roots of Ficus microcarpa. The new synthetic method

of salvinal (23) was shown as in Fig.1 . Fractionation and identification of 9c, 11t, 13

t -conjugated linolenic acid (24) as an

activator of PPARα in bitter gourd (Momordica charantia L.)

四、、、Published papers、Published papersPublished papers Published papers

1. Kuo, Y. C., Y. H. Kuo, Y. L. Lin, and W. J. Tsai, Antivir. Res., 70(1), 112-120 (2006).

2. Hsia S. M., W. C. Chiang, Y. H. Kuo, and P. S. Wang, Int. Impot. Res., 18 (1), 264- 274 (2006).

3. Hsieh, C. L., L. L. Shiu, M. H. Tseng, Y.Y. Shao and Y. H. Kuo, J. Nat. Prod.

69(3), 665-667(2006).

4. Chen, C. C., Y. J. Shio, R. D. Lin, Y. Y. Shao, M. N. Lai, C. C. Lin, L. T. Ng, and Y. H. Kuo, J. Nat. Prod., 69(3), 689-691(2006).

5. Lee, T. H., M. H. Yeh, C. I. Chang, C. K. Lee, Y. Y. Shiao, and Y. H. Kuo, Chem.

Pharm. Bull., 54(5), 693-695(2006).

6. Hsieh, C. L, M. H. Tseng,Y. Y. Shao, J. Y. Chang, C. C. Kuo, C. Y. Chang, and Y. H. Kuo, J. Nat Prod., 69(11) , 1611-1613(2006).

7. Ouyang, M. A.and Y. H. Kuo, J. Asian.

Nat. Prod. Res., 8(7),625-630 (2006).

8. Wang ,E. C.,Y. S.Wein, and Y. H. Kuo,

Tetrahedron Lett. , 47,9195-9197(2006).

9. Chuang, C. Y., C. Hsu, C. Y. Chao, Y. S. Wein, Y. H. Kuo, and C. J. Huang, J.

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MeO OMe OMe O O O O 1 12 13 14 15 16 20 21 22 OH H OH H H H OH H H H OH H H 4 5 6 7 8 ₉ 10 11 17 17a 18 2 3 19 5

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23 Fig.1 COOH 9 10 11 24 6

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1

第十二屆分子科學學會年會

〈Society for

Biomolecular Sciences 12

th

Annual Conference &

Exhibition〉

郭悅雄教授 (Yueh- Hsiung Kuo)

國立台灣大學化學系

計畫編號 NSC-95-2113-M-002-004

一、參加會議

此屆大會是第十二屆分子科學學會年會，此學會是世界

性組織，每年在不同地區舉辦，邀請世界頂尖研究學者作邀

請演講。此屆在美國西雅圖世界貿易中心舉辦 (Washington

State Convention & Trade Center)。西雅圖市是美國十大合宜

人居住的地方，人口不多，古木蒼天，緯度甚高，但下雪的

日子不多，有一個華盛頓湖，就約佔全市的二分之一大，氣

候宜人景色優美。此次舉辦重點是

Advancing Drug Discovery:

From Better Hits to Better Candidates。藥物研究是人類的未

來，因此此次大會參加人數甚多，Plenary Letter 有 8 位，

workshorp 6 位 Short Lectures 26 位，並有數百篇的壁報論

文，在整個世貿中心，展示三天。此次大會的重點有多項：

1. ADME/Toxicology in Early Drug Discovery. 2. Advanced

Technology for HTS. 3. Biomarkers - Preclincal & Clinical Eval.

New Drugs. 4. Ceating Novel Small Molecule Lead Compound.

5. Defining Target & Compound Specificity. 6. Drug

Discovery – Diseases of the Developing World. 7. Strcuture

Biology & Lead Optimization。大會共有五天，由 9 月 17 日

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2

到

21 日，17 日起開始登記，晚上有 receiption 晚會，18 日

起早上有主演講及口頭報告，而壁報論文三天連續。

二、與會心得

藥物研發是著者的主要工作，故此

section 是著者最感興

趣，New Frontiers in Medicinal Chemistry 之演講者大部分都

來自美國藥廠的研發主持人。台灣的藥物研發還在初期學習

中，台灣的藥廠規模只能算是小企業，所以學術界經二十年

的研究，台灣還是沒有半個要能進

Phase 1 的藥物，而致於

臨床藥更不用說，因台灣合成藥物可以說是一個實驗室作單

打獨鬪，要合成出二百多種衍生物，須花費十年時間，又藥

理技術平台缺乏，亦少人從事分子模擬，無法找出最合適的

設計藥物，如此可省去非常多時間。此會最大的收穫是更深

入了解藥物研發的程序，藥物可能來自合成的，也可能來自

天然的，再經合成或修飾去作

optimization，再由分子模擬的

computer，設計新化合物，最後用 high thrughput 的工具做活

性測試，而合成化合物是上一千、二千以上，才能找到最佳

藥物，非台灣可能望其項背，以台灣最大規模藥廠神隆及泰

景都尚未能新藥上

Phase 1 的藥物，其他學術單位更不用說。

此次討論會中，有

Malaria 的新藥發現。Antithrombotics 之藥

物，用的是

hexasaccharide sulfate。Prof. Ulrich 提出 Epothilones

之合成具有

cytostatics activity。PPAR

α

，PPAR

β

是在

lipid 與

glucose 之代謝有用的酵素，而 Prof. Azimioara 合成 scaffolds

作為

PPAR

β

的活化物。NO 之合成有三種 enzyme 分別是

e-NOS、n-NOS、及 i-NOS，而 i-NOS 產物的 NO 會致使發

炎、關節炎、及

osteoarthiritis，而 i-NOS 為 135KDa 之蛋白

質，設計一些

pyridine-pipyrroline 之化合物其 IC

50

可低到

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3

常

e-NOS 沒影響，是最佳抗發炎的藥物。

參與本會的台灣學者甚少，本人以 Synthesis and

Evalution of Cytotoxic Taiwanin A derivatives 之題目發表，

Taiwanin A 是一個 dibenzylidene butyrolactone 木酚素，僅出

現於台灣杉中，它對

A-549, MCF-7 及 HI-29 之癌細胞之抑

制活性

IC

50

為

0.4 μM, 0.5μM 及 0.3μM，經由 apoptosis，而

對

Hep A2, Hep 3B/T2 之活性為 EC

50

為 0.25 及 1.00 μM，經

由

G2/M phase arrest ，它與臨床藥 nocodazole 相似，對 cell 之

microtube 為 destabilize。此次由 Benzaldehyde 進行合成 10

種

Taiwanin A 衍生物。以 HONE-1 Cell 作評估，以 Taiwanin

A 及其中四種衍生物發現其 IC

50

為

4.5μM (Taiw. A). 3.8μM,

4.4μM, 2.6μM 及 0.5μM。如 Taiwanin A 以 TsOH 在 THF 中

不見光，可得

1-phenyl-1,4- dihydronaphthalene，其對 HONE-1

之

IC

50

為 0.28μM 是 Taiwanin A 之 16 倍，此化合物是值得

發展。

三、建議

一種國際性會議是集合世界先進國之工作發表會，可了

解最新研究的動向與情報，要鼓勵國內學者多參與國際性研

討會，可快速獲得最新有關自己研究的資料，建議國內各有

關單位如想獲得國外學者智慧結晶，則多舉辦一些不需要很

大的國際會議，可提昇自己些研究的水準。

四、攜回資料

2007 年 SBS 12

th Annual Conference & Exhibition 之

Abstract book。

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1 Yueh-Hsiung Kuo

1

, Jang-Yang Chang

2

, Ching-Chuan Kuo

2

, and Yung-Shung Wein

1

_{Department of Chemistry, National Taiwan University, Taipei, Taiwan, ROC}

2

_{Division of Cancer Research, National Health Research Institutes, Taipei, Taiwan}

Synthesis and Evaluation of

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2 Introduction

Taiwanin A, a dibenzylidene butyrolactone lignan, exists only in Taiwania

cryptomerioides. The previous paper, we have reported the strong cytotoxic

activity against three human tumor cells, A-549, MCF-7, and HT-29 with IC

₅₀

values 0.4µM, 0.5µM, and 0.3µM, respectively. In addition, the

morphology-based evaluation, flow cytometric analysis, and DNA fragmentation assays

demonstrated that the tumor cell death induced by Taiwanin A was due to

apoptosis. In another research, two hepatoma cells, Hep A2, Hep 3B/T2 were

treatment with Taiwanin A, and found that Taiwanin A arrested the hepatoma

cell at G2/M phase (IC

₅₀

0.25 and 1.00µM, respectively). Taiwanin A might be

mimic nocodazole to destabilize microtubule of the cell. Benzaldehyde

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3

O O O O O O Taiwanin A (1)

Cytotoxicity of 1 isoloated from Taiwania cryptomerioides (IC

50

μM)

Cell – line

A-549 MCF-7 H-T-29 Hep

A2 Hep

3B/T2

1 0.4 0.5 0.3 0.25 1.00

Adriamycin 0.02 0.1 0.1

Taxol

0.15 1.20

Chang, S. T., Kuo, Y. H. et al. Phytochemistry, 2000, 55, 227-232.

Taiwanin A (1) can be prepared by two methods ( A and B methods).

B Method exhibited higher yield.

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4 Preparation of 1 by A Method

O O CHO O O O O HO OH O O

1) NaH / C

₆

H

₅

CH

₃

/ /△

EtO

2

C

CO

2

Et

2) acidify with 10% HCl

O O O O O O O

excess eq. AcCl / C

₆

H

₅

CN

reflux

1~2 drops H

_{MeOH / reflux}

2

SO

4

untill to no 4

3

4

2 ( 35 %)

( 30 %)

( 65 %)

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5 Cytotoxicity of 6 ~ 15 (IC

₅₀

μM)

R Hep G2/A2 Hep 3B/T2

6

Me 12.1 8.9

7

Pr 12.2 7.6

8

Bu 10.8 6.5

9

(CH2)4CH3 10.4 2.0

10

(CH2)5CH3 20 6.0

11

(CH2)7CH3 > 20 1.8

12

CH2Ph > 20 5.7

13

(CH2)3Ph 8.5 9

14

CHMe2 2.8 6.6

15

CH2CHMe2 1.8 9 O O O O RO OR O O O O O O O O

ROH , H

+

/ C

₆

H

₆

reflux

6~15

Taiwanin A (1)

O O O O OMe OH O O

5 1.4 eq. DIBAL / CH

₂

Cl

₂

- 78 ℃ 0℃

2) 10%HCl pH 1

2 ( 20 %)

( 50 %)

+ others

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6

O O O O OH HO

16 1) DIBAL / CH

₂

Cl

₂

, rt

2) 10%HCl pH 1

6

O O O O H H O O

17 PCC

Hep G2/A2 (IC

₅₀

1.0μM)

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7 Preparation of 1 derivatives by B Method

CHO R1 R2 R3 + MeO _OMe O O NaOMe MeOH, reflux 16 h MeI, acetone K2CO3, reflux 8 h CO2Me CO2Me R1 R2 R3 18 R1=R3=H, R2=OCH3(82%) 19 R1,R2=OCH2O, R3=H (81%) 20 R1=R2=R3=OCH3(80%) CHO R3' R2' R1'

NaOMe, MeOH, reflux, 8 h

O OMe O -R₁ R2 R₃ R1' R2' R₃' MeO₂C O MeO2C R1 R2 R3 R₁' R2' R3' O MeO -H H H + R1 R2 R2' R3' HO2C CO2Me R3R1' 21 R1=R3=H, R2=OCH3, R1'=H,R2',R3'=OCH2O (87%) 22 R1=R3=H, R2=OCH3, R1'=R2'=R3'=OCH3 (85%) 23 R1,R2=OCH2O,R3=H, R1'=R3'=H,R2'=OCH3 (88%) 24 R₁,R₂=OCH₂O,R₃=H, R₁'=R₃'=H,R₂'=NO₂ (84%) 25 R1,R2=OCH2O,R3=H, R1'=H,R2'=R3'=OCH3 (85%) 26 R1,R2=OCH2O,R3=H, R1'=R2'=R3'=OCH3 (86%) 27 R1=R2=R3=OCH3, R1'=R3'=H,R2'=OCH3 (86%) 28 R1=R2=R3=OCH3,R1'=H,R2',R3'=OCH2O (88%) 29 R₁=R₂=R₃=OCH₃,R₁'=OCH₃,R₂',R₃'=OCH₂O (82%) 30 R1=R2=R3=OCH3,R1'=R2'=R3'=OCH3 (81%) LiEt3BH THF ClCOOEt Et3N, THF R1 R2 R2' R3' R3R1' O

O 32 31 RR1₁=R=R3₃=H, R=H, R2₂=OCH=OCH₃3, R, R1₁'=R'=H,R₂'=R2',R₃'=OCH3'=OCH₃ (78%)2O (80%) 33 R1,R2=OCH2O,R3=H, R1'=R3'=H,R2'=OCH3 (76%) 34 R1,R2=OCH2O,R3=H, R1'=R3'=H,R2'=NO2 (86%) 35 R1,R2=OCH2O,R3=H, R1'=H,R2'=R3'=OCH3 (78%) 36 R1,R2=OCH2O,R3=H, R1'=R2'=R3'=OCH3 (75%) 37 R1=R2=R3=OCH3, R1'=R3'=H,R2'=OCH3 (77%) 38 R1=R2=R3=OCH3,R1'=H,R2',R3'=OCH2O (79%) 39 R1=R2=R3=OCH3,R1'=OCH3,R2',R3'=OCH2O (83%) 40 R1=R2=R3=OCH3,R1'=R2'=R3'=OCH3 (76%)

(18)

8

O O R1 O O 1 R1 = H, R2 + R3 = OCH2O 38 R1 = R2 = R3 = OCH3 O O O R3 O O O O R3 O OH H+ O O O OH O O O O 1_O 2 ene reaction O O O R3 O O O _H H H O O O R3 O O tautomerization H R3 R2 _R 2 R3 R1 R2 R3 R1 R2 R1 R1 R2 R2 R1 R2 R₁ 1_O 2 , CHCl3 , hv -40℃, TPP 41 R1 + R2 = OCH2O, R3 = H ( 62%, Taiwanin C ) 43 R1 = R2 = R3 = OCH3 ( 60% ) 42 R1 + R2 = OCH2O, R3 = H ( 35%, Taiwanin E ) 44 R1 = R2 = R3 = OMe ( 29% )

Taiwanin C (41) strongly suppressed the production of HBsAg (HBV)

by both Hep A2 and Hep 3B/T2 cells with IC

₅₀

of 0.5μg/ml and 0.4μg/ml,

respectively . Taiwanin E (42) exhibited no effect.

(19)

9

O O O O O O

taiwanin A (1)

H+ O O O O O O

Dihydrotaiwanin C (45) ( 73% )

O O O O O OH tautomerization H O O O O O O H deprotonation then

( IC

₅₀

0.28 μM to HONE - 1

)

(20)

10

O O MeO MeO MeOH, TsOH PtO₂.H₂O H₂ O O MeO MeO OMe O O OMe O O

38

46 (95%)

O O MeO MeO MeOH, TsOH PtO₂.H₂O H₂

40 ₄₇

_(95%)

OMe OMe OMe MeO O O MeO MeO OMe OMe OMe MeO

(21)

11 Cytotoxic activities of Taiwanin A (

1 ) derivatives were shown as

following ( Toxic against HONE-1 cell ,

1 was used reference)

Ar1 = _O O Ar1 = OMe 1 Ar2 = _O O 4.5 μM 0.1μM (HepG2) 0.07μM(HeLa) 31 Ar2 = O O 18.6μM Ar1 = OMe Ar1 = _O O Ar1 Ar2 O O 32 Ar2 = OMe OMe OMe 22μM 33 Ar2 = OMe 3.8μM Ar1 = _O O Ar1 = _O O 34 Ar2 = NO2 1-10μM(Hep-3B) 35 Ar2 = OMe OMe 4.4μM 10.6μM(HeLa) Ar1 = _O O Ar1 = OMe OMe OMe 36 Ar2 = OMe OMe OMe 18μM 37 Ar2 = OMe 2.6μM 0.32μM(Hep G2) 1.27μM(HeLa)

(22)

12

Ar1 = OMe OMe OMe Ar1 = OMe OMe OMe 38 Ar2 = _O O 0.5μM 39 Ar2 = O O OMe 5.8μM Ar1 = OMe OMe OMe 46 and 47 > 50μM Ar1 = Ph 40 Ar2 = OMe OMe OMe 17μM 48 Ar2 = Ph 24μM (HepG2) 18.1μM(HeLa) O O O O O O O O OH OH O O O O O O O O HO O O O O O OH Savinin (51) Hinokinin (52) ₅₃ ₅₄

(23)

13 Conclusion of SAR

1. γ-Lactone and two double bonds are the essential

functionalities.

2. Compound

17 without γ -Lactone expressed strong activity.

Because the aldehyde is an active functionality.

3. Ar

₁

moiety with three electron-releasing –OR groups

exhibited more activity than two –OR or one –OR groups.

4. The substituents with releasing or

electron-withdrawing group in Ar

₂

did not affect the cytotoxic activity.

5. Less steric hindrance in Ar

₂

moiety showed the higher

activity (

38 >

39 >

40 )

6. Phenyl 1,4-dihydronaphalene derivative (45) unexpectedly

presents strong cytotoxic activity.

(24)

1

第十二屆分子科學學會年會

〈Society for

Biomolecular Sciences 12

th

Annual Conference &

Exhibition〉

郭悅雄教授 (Yueh- Hsiung Kuo)

國立台灣大學化學系

計畫編號 NSC-95-2113-M-002-004

一、參加會議

此屆大會是第十二屆分子科學學會年會，此學會是世界

性組織，每年在不同地區舉辦，邀請世界頂尖研究學者作邀

請演講。此屆在美國西雅圖世界貿易中心舉辦 (Washington

State Convention & Trade Center)。西雅圖市是美國十大合宜

人居住的地方，人口不多，古木蒼天，緯度甚高，但下雪的

日子不多，有一個華盛頓湖，就約佔全市的二分之一大，氣

候宜人景色優美。此次舉辦重點是

Advancing Drug Discovery:

From Better Hits to Better Candidates。藥物研究是人類的未

來，因此此次大會參加人數甚多，Plenary Letter 有 8 位，

workshorp 6 位 Short Lectures 26 位，並有數百篇的壁報論

文，在整個世貿中心，展示三天。此次大會的重點有多項：

1. ADME/Toxicology in Early Drug Discovery. 2. Advanced

Technology for HTS. 3. Biomarkers - Preclincal & Clinical Eval.

New Drugs. 4. Ceating Novel Small Molecule Lead Compound.

5. Defining Target & Compound Specificity. 6. Drug

Discovery – Diseases of the Developing World. 7. Strcuture

Biology & Lead Optimization。大會共有五天，由 9 月 17 日

(25)

2

到

21 日，17 日起開始登記，晚上有 receiption 晚會，18 日

起早上有主演講及口頭報告，而壁報論文三天連續。

二、與會心得

藥物研發是著者的主要工作，故此

section 是著者最感興

趣，New Frontiers in Medicinal Chemistry 之演講者大部分都

來自美國藥廠的研發主持人。台灣的藥物研發還在初期學習

中，台灣的藥廠規模只能算是小企業，所以學術界經二十年

的研究，台灣還是沒有半個要能進

Phase 1 的藥物，而致於

臨床藥更不用說，因台灣合成藥物可以說是一個實驗室作單

打獨鬪，要合成出二百多種衍生物，須花費十年時間，又藥

理技術平台缺乏，亦少人從事分子模擬，無法找出最合適的

設計藥物，如此可省去非常多時間。此會最大的收穫是更深

入了解藥物研發的程序，藥物可能來自合成的，也可能來自

天然的，再經合成或修飾去作

optimization，再由分子模擬的

computer，設計新化合物，最後用 high thrughput 的工具做活

性測試，而合成化合物是上一千、二千以上，才能找到最佳

藥物，非台灣可能望其項背，以台灣最大規模藥廠神隆及泰

景都尚未能新藥上

Phase 1 的藥物，其他學術單位更不用說。

此次討論會中，有

Malaria 的新藥發現。Antithrombotics 之藥

物，用的是

hexasaccharide sulfate。Prof. Ulrich 提出 Epothilones

之合成具有

cytostatics activity。PPAR

α

，PPAR

β

是在

lipid 與

glucose 之代謝有用的酵素，而 Prof. Azimioara 合成 scaffolds

作為

PPAR

β

的活化物。NO 之合成有三種 enzyme 分別是

e-NOS、n-NOS、及 i-NOS，而 i-NOS 產物的 NO 會致使發

炎、關節炎、及

osteoarthiritis，而 i-NOS 為 135KDa 之蛋白

質，設計一些

pyridine-pipyrroline 之化合物其 IC

50

可低到

(26)

3

常

e-NOS 沒影響，是最佳抗發炎的藥物。

參與本會的台灣學者甚少，本人以 Synthesis and

Evalution of Cytotoxic Taiwanin A derivatives 之題目發表，

Taiwanin A 是一個 dibenzylidene butyrolactone 木酚素，僅出

現於台灣杉中，它對

A-549, MCF-7 及 HI-29 之癌細胞之抑

制活性

IC

50

為

0.4 μM, 0.5μM 及 0.3μM，經由 apoptosis，而

對

Hep A2, Hep 3B/T2 之活性為 EC

50

為 0.25 及 1.00 μM，經

由

G2/M phase arrest ，它與臨床藥 nocodazole 相似，對 cell 之

microtube 為 destabilize。此次由 Benzaldehyde 進行合成 10

種

Taiwanin A 衍生物。以 HONE-1 Cell 作評估，以 Taiwanin

A 及其中四種衍生物發現其 IC

50

為

4.5μM (Taiw. A). 3.8μM,

4.4μM, 2.6μM 及 0.5μM。如 Taiwanin A 以 TsOH 在 THF 中

不見光，可得

1-phenyl-1,4- dihydronaphthalene，其對 HONE-1

之

IC

50

本土杉科與柏科植物成分及其活性之研究(2/2)

行政院國家科學委員會專題研究計畫 成果報告