壹、設計與合成2-胺基-3,4,5-三甲氧基二苯甲酮作為強效微管蛋白聚合抑制劑 貳、設計與合成聯苯基苯磺醯胺衍生物作為新穎抗癌試劑之合成和結構與活性關係研究 參、設計與合成2-胺基苯醯胺之苯磺醯胺衍生物作為強效抗癌試劑

I. Design and Synthesis of 2-Amino-3,4,5-Trimethoxybenzophenones as Potent Tubulin Polymerization Inhibitors II. Design and Synthesis of Biphenylbenzenesulfonamide Derivatives as Novel Anticancer Agents III. Design and Synthesis of Benzenesulfonamides with 2-Amino- phenylamide as Potent Anticancer Agents

莊勳岳(Hsun-Yueh Chuang)

指導教授 ： 劉景平

臺北醫學大學/藥學院/藥學系(博士班)/博士(2012年)

摘要

壹、設計與合成2-胺基-3,4,5-三甲氧基二苯甲酮作為強效微管蛋白聚合抑制劑

二苯甲酮類結構衍生物如phenstatin，具有良好抑制微管聚合及抑制癌細胞之活性。二苯甲酮結構中酮基構形使兩芳香環間的位向，與combretastatin A-4 (CA-4)活性所必需之順式stilbene位向相似，且與順式stilbene相比二苯甲酮結構較易於化學合成及控制結構位向。另外本實驗室之經驗於CA-4之A環C-2位導入胺基能增加化合物之極性並維持抑制微管聚合之活性。本論文設計與合成一系列新穎之2-胺基-3,4,5-三甲氧基二苯甲酮衍生物，並對五種人類癌細胞進行細胞株毒殺測試包括口腔上皮細胞癌細胞(KB cells)、大腸直腸癌細胞(HT29 cells)、非小細胞肺癌細胞(H460 cells)、胃癌細胞(MKN45 cells)和多抗藥性癌細胞(KB-vin10)，顯示具有良好抑制癌細胞之活性(IC50值範圍為7~36 nM)。化合物292、296及297，由微管蛋白競爭性試驗發現具有顯著的抑制微管聚合活性(IC50分別為1.7、2.0及1.6 μM)，與CA-4 (IC50 = 1.9 μM)相當。由實驗結果證明2-胺基-3,4,5-三甲氧基二苯甲酮化合物對於抑制微管聚合具有強抑制活性可研究發展做為新穎微管聚合抑制劑之抗癌藥物。

貳、設計與合成聯苯基苯磺醯胺衍生物作為新穎抗癌試劑之合成和結構與活性關係研究

目前許多苯磺醯胺結構之化合物發現具有抗癌活性且已進入臨床試驗，本論文根據ABT-751之苯磺醯胺結構，設計與合成以聯苯基苯磺醯胺為骨架之一系列化合物。對四種人類癌細胞進行生長抑制實驗，結果顯示此系列化合物將A環C-4位以拉電子基(苯腈)取代(319)，具有良好的癌細胞生長抑制活性，而將A環C-4取代基改為推電子基及較大基團取代或改至A環C-3位置或改為A環芳香雜環，抑制活性明顯下降甚至喪失。此外同時於A環C-3位置與C-4位置導入拉電子基團(3-氟及4-苯腈，331)具有最佳癌細胞抑制活性(GI50值範圍為0.02~0.10 μM)。另外進一步對化合物319進行修飾，將苯磺醯胺改為三級胺、醯胺或亞甲基苯磺醯胺，活性均明顯下降或喪失，證明苯磺醯胺結構對抗癌活性相當重要，聯苯基苯磺醯胺對於抑制癌細胞生長具有良好抑制活性可進一步研究發展做為有效之抗癌試劑。

參、設計與合成2-胺基苯醯胺之苯磺醯胺衍生物作為強效抗癌試劑

根據文獻回顧，許多進入臨床試驗藥物具有良好的組蛋白去乙醯酶抑制活性如MS-275及MGCD-0103，皆具有2-胺基苯醯胺結構，另外本論文第一部份研究發展出苯磺醯胺衍生物具有良好癌細胞抑制活性。因此，本論文結合2-胺基苯醯胺與苯磺醯胺結構，合成一系列創新結構之化合物。以吡啶苯磺醯胺為主結構，鄰位以胺基亞甲基連結2-胺基苯醯胺之化合物428，活性結果顯示對口腔癌細胞(KB cells)具有良好的抑制活性(IC50為45.5 nM)。若將化合物428改為苯基苯磺醯胺(434)其抗癌活性顯著降低。以苯基苯磺醯胺(434)為主結構，鄰位以不具極性且非固定構型之C-C單鍵連結(437)，抗癌活性顯著增加(IC50 = 6.2 nM)。其他化合物目前正在進行活性測試。由初步實驗結果顯示2-胺基苯醯胺之苯磺醯胺衍生物(437)對於抑制癌細胞生長具有良好抑制活性，可進一步研究發展為強效之抗癌試劑。

關鍵字

抗微管劑 ； 二苯甲酮 ； 抑制劑 ； 聚合 ； 微管

並列摘要

I. Design and Synthesis of 2-Amino-3,4,5-Trimethoxybenzophenones as Potent Tubulin Polymerization Inhibitors

Benzophenone derivatives, such as phenstatin, exhibited substantial antimitotic and antitumor activities. The carbonyl group of benzophenone links the two aryl rings in a quasi cis-stilbene, which appears to be necessary for significant biological activity of combretastatin A-4 (CA-4), and the benzophenone compounds are easily synthesized than the cis-stilbenes, which require control of geometric selectivity. Our previous reports that the amino group substituent at the 2-position of ring A in the CA-4 increased the polarity of the corresponding structure without compromising bioactivity. This dissertation presented that the design and synthesis of a novel of 2-amino-3,4,5-trimethoxybenzophenone analogues displayed remarkable anticancer activity, with IC50 values ranging from 7 ~ 16 nM against five human cancer cell lines including KB cells, HT29 cells, H460 cells, MKN45 cells and KB-vin10. Compounds 292, 296 and 297 exhibited excellent inhibitory of tubulin polymeriztion by targeting the colchicine binding site of microtubules (IC50 = 1.7, 2.0, 1.6 μM, respectively), similar to that of CA-4 (IC50 =1.9 μM). Overall, the 2-amino-3,4,5-trimethoxy- benzophenones exhibited strong inhibition of tubulin polymerization has potential for further investigation as anticancer agents.

II. Design and Synthesis of Biphenylbenzene-sulfonamide Derivatives as Novel Anticancer Agents

Lots of sulfonamide derivatives have been found potent anticancer activities and are undergoing clinical trials. This dissertation presented the design and synthesis of a series of biphenylbenzensulfonmaide derivatives based on the benzensulfonmaide structure core of ABT-751. Compounds bearing electron withdrawing groups (cyano group) at 4-position in ring A (319) displayed remarkable anticancer activity. However, electron donating groups and larger functional groups at 4-position, substitution at 3-positon in ring A, or changed the ring A with heteroaromatic ring, exhibited dramatic decrease or loss anticancer activities. Additionally, compound 331 with electron withdrawing groups, 3-fluoro and 4-cyano group in ring A, displayed excellent anticancer activities with mean GI50 values ranging from 0.02~0.10 μM against four human cancer cell lines. We further investigated the activities of benzensulfonamide by modifying compound 319. The anticancer activities were significant decrease or loss by changing the benzenesulfonamide to tertiary sulfonamide, amide or methylenesulfonamide. The structure activity relationship information suggested that the benzenesulfonamide moiety may play an important role in the activity of this series of compounds, and biphenylbenzensulfonmaide, exhibiting good inhibition of cancer cell growth has potential for further investigation as an anticancer agent.

III. Design and Synthesis of Benzenesulfonamides with 2-Aminophenylamide as Potent Anticancer Agents

Based on the literature surveys, 2-aminophenylamide derivatives such as MS-275 and MGCD-0103, display good inhibition of HDAC and are undergoing clinical trials. And the benzenesulfonamide derivatives exhibited good anticancer activities which showed in part I of this dissertation. This dissertation presented a novel series of compounds combined 2-aminophenylamide moiety with benzenesulfonamide. Compound 428 with ortho-methyleneamine linker between pyridinylbenzene- sulfonamide and 2-aminophenylamide displayed good anticancer activity against KB cells (IC50 = 45.5 nM). The activity was significant decreased by replacing the pyridine of compound 428 with benzene ring (434). We further modified the compound 434 to compound 437 with a non-polar and non-rigid C-C bond linker, and the activity was dramatic increased (IC50 = 6.2 nM). The cell growth inhibitory assay of other compounds is in progress. According to the data at present, benzenesulfonamides with 2-aminophenylamide (437) displaying excellent anticancer activity has potential for further investigation as an anticancer agent.

並列關鍵字

antimitotic agent ； benzophenones ； inhibitors ； polymerization ； tubulin

參考文獻

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