I. Design, synthesis and anticancer activity of 2-(3-Benzo[b]thienyl)-6,7-methylenedioxyquinolin-4-one analogs.II. Design, synthesis and biological activity of 4-phenyl-1H-benzo[b][1,4]diazepin-2(3H)-one derivatives.

• Main Authors: Yu-Hsun Chang, 張昱勛
• Other Authors: 郭盛助
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/44593742482148797794

博士 === 中國醫藥大學 === 藥物化學研究所博士班 === 97 === Part I. As part of our continuing investigation of azo-flavonoid derivatives as potential anticancer drug candidates, a series of 2-aryl-6,7-methylenedioxyquinolin-4-one analogs was designed and synthesized. The design combined structural features of 2-(2-fluorophenyl)-6,7-methylenedioxyquinolin-4-one (CHM-2133), a previously discovered compound with potent in vivo antitumor activity, and 2-arylquinolin-4-ones identified by CoMFA models. The newly synthesized analogs were evaluated for cytotoxicity against several human cancer cell lines, and structure-activity relationship (SAR) correlations were established. Analogs 1, 37, and 39 showed potent cytotoxicity against different cancer cell lines. Compound 1 demonstrated selective cytotoxicity against Hep 3B (hepatoma) cells. Compound 37 was cytotoxic against HL-60 (leukemia), HCT-116 (colon cancer), Hep 3B (hepatoma), and SK-MEL-5 (melanoma) cells. Compound 39 exhibited broad cytotoxicity against all seven cancer cell lines, with IC50 values between 0.07−0.19 μM. Results from mechanism of action studies revealed that these new quinolone derivatives function as anti-tubulin agents. Continuing with the screening result, CWC-8 (compound 39) was chosen to detect the mechanism of anticancer activity in the experiment. We have defined the viability inhibition and apoptotic mechanisms of CWC-8 on human osteogenic sarcoma U-2 OS cells. According to the MTT assay, the cell viability was inhibited by CWC-8 in a dose- and time-dependent manner, with an IC50 of 4.97±0.24 μM. CWC-8 treatment induced G2/M arrest and apoptosis in U-2 OS cells by cell cycle and flow cytometry analysis. Western blotting and CDK1 kinase assay showed that CWC-8 treatment caused a time-dependent increase of Cyclin B and CDK1 protein levels and activity during G2/M arrest. CWC-8 treatment also caused a time-dependent increase in Fas/CD95, FADD, cytosolic cytochrome c, caspase-8/-9/-3 active form, Apaf-1, AIF, Bax protein levels, and decrease in Bcl-2 protein level. CWC-8 also promoted caspase-8/-9 and -3 activities; however, pretreatment of cells with pan-caspase, caspase-8/-9 and -3 inhibitors led to reduced cell growth inhibition action. Taken together, these findings show CWC-8 could be a potential candidate for cancer therapy. Part II. The intrinsic mobility of proteins has often been ignored in drug design field. Conformational induction is the energy balance process in which ligand converts protein into a conformation that would not spontaneously adopt in its unligated state. This flexible property of protein limits the development of rational drug design model nowadays. Therefore, we proposed a concept that describes raise of ligand flexibility is the strategy to accommodate protein mobility. In this study, we synthesized a series of 4-phenyl-1H-benzo[b][1,4]diazepin-2(3H)-one derivatives which are more flexible than our previous antitumor compound− 2-phenyl-4-quinolones. Unexpectedly, the cytotoxicity screening result shows little activity of these derivatives. Only 7-4, 7-5, 7-9, 7-15, 7-16, 7-20 and 7-27 compounds exhibt the potency to inhibt HL-60 cell line. Despite the unideal outcome in anti-cancer test, we acquired the anti-inflammation activity of compound 7-4 via random screening insteadly. It inhibited fMLP-induced superoxide anion production through a PKA-dependent pathway and increased cAMP by activating protein phosphatase 2A, which subsequently inhibited phosphodiesterase 4.