資料載入中.....
|
請使用永久網址來引用或連結此文件:
http://ir.cmu.edu.tw/ir/handle/310903500/904
|
| 題名: | 2-[氮-多樣取代苄基]取代苯胺基-4-酮基-4,5-二氫呋喃-3-羧酸乙酯類衍生物之合成及其生理活性;Synthesis and Biological Activity of Certain Ethyl 2-[N-various Substituted benzyl]Substituted anilino-4-oxo-4,5-dihydrofuran-3-carboxylate |
| 作者: | 黃安正;An-Cheng Huang |
| 貢獻者: | 中國醫藥大學：藥物化學研究所博士班 |
| 關鍵詞: | 2-取代苯胺-4-酮基-4,5-二氫呋;喃-3-乙酸乙酯化合物;2-[氮-多樣取代苄;基]取代苯胺基-4-酮基-4,5-二氫呋;喃-3-羧酸乙酯類衍生物;氮-多樣取代苄;基-7-甲氧基-2,3,4,9-四氫呋;喃并[2,3-b]喹;啉;-3,4-二酮化合物;氮-多樣取代苄;基-7-甲氧基-4,9-二氫呋;喃并[2,3-b]喹;啉;-4-酮化合物;細胞凋亡;活性氧化物;小鼠血癌細胞WEHI-3 cells;人類子宮頸鱗狀上皮癌細胞-Ca Ski cells;ethyl 2-(substituted anilino)-4-oxo-4,5-dihydrofuran-3-carboxylate;2-[N-various benzyl] substituted anilino-4-oxo-4,5-dihydro-furan -3-carboxylate;N-various benzyl-7-methoxy-2,3,4,9-tetrahydrofuro- [2,3-b]-quinolin-3,4-diones;N-various benzyl-7-methoxy- 4,9-dihydrofuro[2,3-b]quinolin-4-diones;apoptosis;reactive oxygen species;human cervical cancer cell line (Ca Ski) |
| 日期: | 2007-01-18 |
| 上傳時間: | 2009-08-13 09:37:13 (UTC+8) |
| 摘要: | 著者合成了一系列2-取代苯胺-4-酮基-4,5-二氫呋喃-3-乙酸乙酯化合物(1-10)、多系列2-[氮-多樣取代苄基]取代苯胺基-4-酮基-4,5-二氫呋喃-3-羧酸乙酯類衍生物(11~151)、一系列氮-多樣取代苄基-7-甲氧基-2,3,4,9-四氫呋喃并[2,3-b]喹啉-3,4-二酮化合物(155~167)、一系列氮-多樣取代苄基-7-甲氧基-4,9-二氫呋喃并[2,3-b]喹啉-4-酮化合物(169~180)及相關衍生物，並經各種圖譜資料予以確認結構正確，將所有合成經圖譜確認結構之化合物分別測試其生理活性。
在抗癌活性方面，以小鼠血癌細胞WEHI-3 cells之存活率試驗來評估。從目前測試結果發現Ethyl 2-[N-benzyl-(3’-methoxy)]anilino-4-oxo-4,5-dihydrofuran-3-carboxylate (11)、Ethyl 2-[N-p-fluoro-benzyl-(2’-methyl)]anilino-4-oxo-4,5-dihydrofuran-3-carboxylate (83)、N-m-chloro- benzyl-7-methoxy-2,3,4,9-tetrahydrofuro[2,3-b]-quinolin-3,4-dione (160)、N-o-methoxybenzyl-7- methoxy-4,9-dihydrofuro[2,3-b]-quinolin-4-dione (177)及N-o-chlorobenzyl-7-methoxy-4,9- dihydrofuro[2,3-b]quinolin-4-dione (172)對於WEHI-3 cells之存活率有較大的抑制影響，其1µM 時，WEHI-3 cells之存活率皆低於3%。
對於癌細胞凋亡之研究，選活性差異明顯者進行研究，首先以Ethyl 2-[N-p-chlorobenzyl- (2’-methyl)] anilino-4-oxo-4,5-dihydrofuran-3-carboxylate (80)研究其對人類子宮頸鱗狀上皮癌細胞-Ca Ski cells引起細胞凋亡之機轉，實驗結果顯示化合物80會增加Bax/Bcl-2比值、降低粒腺體膜電位，誘導增加細胞質內Ca2+離子濃度、增加caspases酵素的活化、DNA裂解和細胞凋亡。流式細胞儀也證實化合物80會引起降低粒腺體膜電位。西方墨點法(Western Blotting)顯示化合物80會增加p53、p21、cytochrome c、caspase-3及Bax的蛋白質表現，會降低Bcl-2的蛋白質表現。另由實驗加BAPTA(Ca2+離子螯合劑)，抑制了化合物80所導致細胞質內Ca2+離子濃度的增加、阻止腺體膜電位下降及阻止Ca Ski cells細胞凋亡，證實化合物80誘導Ca Ski cells細胞凋亡是經由與Ca2+離子濃度有關之粒腺體凋亡路徑。
另也研究化合物79誘導人類子宮頸部腫瘤細胞HeLa Cells (簡稱 HeLa Cells)細胞凋亡之機轉。實驗結果顯示化合物79會誘導細胞型態改變和細胞毒性，其作用效果是與劑量呈正向關係。由流式細胞儀偵測法、DAPI 染色法及DNA裂解電泳凝膠法顯示化合物79會誘導HeLa Cells細胞凋亡。由Grp78和Gadd153表現增加，確認化合物79會誘導ROS過量產生，然後造成內質網壓力(ER stress)。西方墨點法證實化合物79會促進p53、Bak、PARP、caspase-3的表現量，另一方面會抑制Bcl-2、Bcl-xL的表現量。化合物79會促進caspase-12的作用，然後導致細胞凋亡。化合物79會抑制MMP-2和MMP-9的表現量，導致抑制HeLa細胞的轉移。
人類嗜中性白血球抗活性氧化物釋放試驗，以fMLP 當發炎反應誘導劑，以Cytochalasin B當細胞軟化劑，提高細胞膜通透性，以Cytochrome c 被H2O2還原成環原態Cytochrome c時，其在波長550之吸光度(OD550)為實驗測量值，以DMSO之(OD550) 為對照組測量值，測量不同濃度藥品對人類嗜中性白血球抗發炎產生ROS之影響。從目前測試結果發現2-(4- chloro-3-oxo-2,3-dihydrofuro[2,3-b]quinolin-8-yl)acetate(183)及N-p-methoxybenzyl-7-methoxy-4,9- dihydrofuro[2,3-b]quinolin-4-dione (180) 對於人類嗜中性白血球之釋放ROS有最大的抑制作用，其IC50值分別為0.022μM 及0.39μM。
其他藥理活性試驗仍在測試中，容待獲得測試結果後，即予以補上，並探討其結構與活性之關係。
One series of ethyl 2-(substituted anilino)-4-oxo-4,5-dihydrofuran-3-carboxylate (1~10), multi-series of ethyl 2-[N-various benzyl] substituted anilino-4-oxo-4,5-dihydro-furan -3-carboxylate (11~151), One series of N-various benzyl-7-methoxy-2,3,4,9-tetrahydrofuro- [2,3-b]-quinolin-3,4-diones (155~167) and another series of N-various benzyl-7-methoxy- 4,9-dihydrofuro[2,3-b]quinolin-4-diones (169~180) have been synthesized and assessed their chemical structures by their spectra data. All of these synthetic compound were evaluated for biological activities.
In anticancer activity, assess with the survival rate of mouse''s blood cancer cell WEHI-3 cells. The test result shows that ethyl 2-[N-benzyl-(3’-methoxy)]anilino-4-oxo-4,5-dihydrofuran-3- carboxylate (11)、ethyl 2-[N-p-fluoro-benzyl-(2’-methyl)]anilino-4-oxo-4,5-dihydrofuran-3- carboxylate (83)、N-m-chloro-benzyl-7-methoxy-2,3,4,9-tetrahydrofuro[2,3-b]-quinolin-3,4- dione (160)、N-o-methoxybenzyl-7-methoxy-4,9-dihydrofuro[2,3-b]-quinolin-4-dione (177) and N-o-chlorobenzyl-7-methoxy-4,9-dihydrofuro[2,3-b]quinolin-4-dione (172)have the greater influence against the survival rate of WEHI-3 cells. In 1µM concentration of above compounds, the survival rate of WEHI-3 cells is all lower than 3%.
Investigation of cancer apoptosis, we conducted Ethyl 2-[N-p-chlorobenzyl-(2’-methyl)]- anilino-4-oxo-4,5-dihydrofuran-3-carboxylate (80) on a human cervical cancer cell line (Ca Ski). In this study, compound 80 acts on a human cervical cancer cell line (Ca Ski) to bring about an increase in the ratio of Bax/Bcl-2, the reduction of the mitochondrial membrane potential (MMP), an increase in the levels of cytoplasmic Ca2+, activation of caspases and fragmentation of DNA, and apoptosis are explored. Flow cytometric analysis demonstrated that compound 80 induced the decline of MMP in Ca Ski cells. Western blotting demonstrated that compound 80 induced an increase in the levels of p53, p21, cytochrome-c, caspase-3 and Bax but a decrease in the level of Bcl-2. compound 80 induced an increase in the level of cytoplasmic Ca2+, which was inhibited by BAPTA (calcium chelator), and BAPTA accelerated the MMP reduction, and significantly blocked compound 80-induced apoptosis. Our data demonstrated that compound 80-induced apoptosis occues via a mitochondria-dependent pathway closely related to the level of cytoplasmic Ca2+ in Ca Ski cells.
We also examined the in vitro effect of compound 79 on human cervical epithelioid carcinoma cell line (HeLa). The results demonstrated that compound 79 induced morphological changes and cytotoxicity in dose-dependent. Compound 79 induced apoptosis which is analyzed by flow cytometric methods and also confirmed by DAPI staining and DNA fragmentation by DNA gel electrophoresis. Compound 79 also induced reactive oxygen species (ROS) overproduction then caused ER stress which also confirmed by the increased levels of Grp78 and Gadd153. Western blotting was selected to demonstrate that compound 79 promoted p53, Bak, PARP, caspase-3 levels and decreased the levels of Bcl-2 and Bcl-xL. Compound 79 also promoted caspase-12 then led to apoptosis. Compound 79 inhibits cell migration of HeLa which is may through inhibition of MMP-2 and 9.
In screen test on reactive oxygen species (ROS) release induced by fMLP in human neutrophil. The test result shows that 2-(4-chloro-3-oxo-2,3-dihydrofuro[2,3-b]quinolin-8-yl) acetate (183) and N-p–methoxy-benzyl-7-methoxy-4,9-dihydrofuro[2,3-b]-quinolin-4-dione (180) have the greatest influence against the reactive oxygen species release induced by fMLP in human neutrophil, its IC50 value is 0.022 µM and 0.39 µM respectively.
Ｍeanwhile, some of these synthetic compounds were still evaluated for their biological activity. Those results of biological activity will be reported later and discussed the relation of its structure and activat |
| 顯示於類別: | [藥物化學研究所] 博碩士論文
|
文件中的檔案:
| 檔案 |
描述 |
大小 | 格式 | 瀏覽次數 |
|---|
| cmu-96-8853005-1.pdf | | 8766Kb | Adobe PDF | 792 | 檢視/開啟 | | index.html | | 0Kb | HTML | 23 | 檢視/開啟 |
|
在CMUR中所有的資料項目都受到原著作權保護.
|
