- 學位論文
建構液相平行合成芬布芬與利尿酸之醯胺分子庫以篩選有效抗癌活性分子之研究
Solution-phase parallel synthesis and screening of anti-tumor activities from fenbufen and ethacrynic acid libraries
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摘要
本研究的重點在於將核心胺基化合物與羧酸化合物以組合化學的方法，在不分離純化的狀態於液相建立起醯胺化分子庫並進行細胞毒性分析。由胺基輔助修飾fenbufen以及ethacrynic acid成為起始核心化合物，再與許多羧酸進行耦合，建立出醯胺化衍生物之分子庫，成功的篩選出了一些有潛力的化合物(N-(4-(4-(biphenyl-4-yl)-4-oxobutanamido) butyl)- 5-(2,5- dimethylphenoxy)-2,2-dimethylpentanamide、N-(4-(4- (biphenyl -4-yl)- 4-oxobutanamide) butyl)-2- ethylhexanamide and 4-(biphenyl-4-yl)- N-(4-(2-(2,3- dichloro-4 -(2-methylenebutanoyl) phenoxy)acetamido) butyl)- 4-oxobutanamide )，可供日後實驗室進行細胞機轉的探討(像是利用Flow cytometry)或是以此化合物為藍本再繼續修飾以獲得SAR(structure and reactivity)方面的藥物機轉研究。 另外，我們也探討了fenbufen、不同碳鏈長之fenbufen醯胺化衍生物及ethacrynic acid醯胺化之衍生物對於巨噬細胞RAW264.7 是否具有細胞存活率的效果，並以MTT 分析法來測試樣品對RAW264.7 巨噬細胞存活率的影響，得知不同碳鏈長之fenbufen醯胺化衍生物及ethacrynic acid醯胺化衍生物濃度高到100 μM 以上時，會顯著地抑制RAW264.7 巨噬細胞的生長，且隨著碳鏈長度的增加，抑制RAW264.7 巨噬細胞的生長的效果越顯著，八個碳鏈長度的fenbufen醯胺化衍生物抑制RAW264.7 巨噬細胞的生長的效果最顯著；還有探討fenbufen、不同碳鏈長之fenbufen醯胺化衍生物及ethacrynic acid醯胺化之衍生物對於LPS 誘發RAW264.7 巨噬細胞產生一氧化氮的影響，結果顯示fenbufen (10 μM)對於NO 的產生也沒有抑制的現象。
並列摘要
Aim of this research is to construct a library screening at a cellular level via amide bond formation under the solution-phase combinatorial chemistry condition without isolation and purification. Amine group and carboxylic acid (fenbufen and ethacrynic acid) were used as raw materials in this research to synthesize N-(4-(4-(biphenyl-4-yl)-4-oxobutanamido) butyl)- 5-(2,5- dimethylphenoxy)-2,2-dimethylpentanamide、N-(4-(4- (biphenyl -4-yl)- 4-oxobutanamide) butyl)-2- ethylhexanamide and 4-(biphenyl-4-yl)- N-(4-(2-(2,3- dichloro-4 -(2-methylenebutanoyl) phenoxy)acetamido) butyl)- 4-oxobutanamide which could be serve as anti-cancers potential compounds. Amide derivatives to establish a molecular library, the success of screening out a number of potential compounds for future laboratory of cellular mechanisms (such as Flow cytometry) or as a blueprint to continue modifying compounds for SAR (structure and reactivity) aspects of drug mechanisms. The previous discoveries of butyl fenbufen amide analogs with antitumor effects were further examined. The amide analogs with 1, 3, 4 and 8 carbons chains were prepared. We probed into the length of the alkyl amide side chain increased, the cytotoxic effects increased, and the octyl fenbufen amide had the greatest cytotoxic effect on RAW 264.7 cells by using the MTT assay. Butyl fenbufen and ethacrynic acid amide analogs exhibited significant cytotoxic effects at a concentration of 100 μM. As the length of the alkyl amide side chain increased, the cytotoxic effects increased, and the octyl fenbufen amide had the greatest cytotoxic effect. In addition, NO in LPS-activated RAW 264.7 cells were measured. At the concentration of 10 μM used in the study, the fenbufen amide analogs did not show cytotoxicity according to the MTT assay results. The NO scavenging activities of the fenbufen amide analogs were not significant different from that of fenbufen.
參考文獻
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