- 學位論文
吡啶[3,2-g]喹啉衍生物之合成及細胞毒活性評估
Synthesis and Cytotoxic Evaluation of Certain Pyrido[3,2-g]quinoline Derivatives
摘要
本研究已合成一系列三環平面吡啶[3,2-g]喹啉(pyrido-[3,2-g]- quinoline)類的衍生物，並進行九大癌細胞六十類癌細胞株之細胞毒性評估；其中化合物9a (diethyl 4,6-diamino-10-methyl- pyrido-[3,2-g]quinoline-3,7-dicarboxylate) 及9d (diethyl- 4,6-bis-(3-dimethylaminopropylamino)-10-methylpyrido[3,2-g]quinoline-3,7-dicarboxylate)對乳癌(MCF-7)、肺癌(NCI-H460)、神經癌(SF-268)及兩種攝護腺癌(LNCaP、PC-3)細胞具有良好的細胞毒活性；進一步評估9a，9b，9c，及9d等化合物對抗乳癌(MCF-7)及兩種攝護腺癌(LNCaP、PC-3)細胞成長的活性，結果顯示降低細胞毒活性的排序為9d > 9a > > 9b及9c；而9d對兩種攝護腺癌細胞(LNCaP、PC-3)及乳癌細胞(MCF-7)之IC50分別為5.63、6.42及3.96 ?戥M；本研究將9d進行細胞週期分析，其結果顯示對9d 抑制LNCaP、MCF-7細胞生長乃由於G1 arrest，隨後並誘發細胞凋亡(apoptosis)；本研究進一步透過Annexin V-FITC及PARP 之裂解模式，確認9d可對LNCaP細胞誘發凋亡。 為了探討9d引發細胞凋亡之機制，本研究進行一些與細胞週期調節有關的蛋白質，包括p53和其下游因子p21及與細胞週期進行有關的cyclin D1、cyclin E、 retinoblastoma (Rb)、phoso-retinoblastoma (pRb)及 c-Myc之研究；結果顯示p53有些微增加，但p21並無變化，此結果表示p21可能不是造成G1 arrest的因子；另外，cyclin D1及cyclin E僅在9d 濃度80 ?戥M時才會減少；相反地，c-Myc、Rb及pRb則隨時間及藥物濃度增加而明顯減少。因此，本研究進一步檢測與c-Myc有關之上游調節因子：Erk1/2和phospho-Erk1/2(p-Erk1/2)，結果發現9d對Erk1/2並無影響，但當9d 濃度5 ?戥M 24小時p- Erk1/2即有增加的現象，顯示9d對LNCaP細胞週期之作用可能是經由p-Erk1/2 路徑影響c-Myc之減少而造成G1 arrest。
並列摘要
The present report describes the synthesis and evaluation of tricyclic pyrido[3,2-g]quinoline derivatives in which an additional pyridine ring is linearly fused on the antibacterial quinoline-3-carboxylic acid. Among them, only both diethyl 4, 6- diamino -10- methylpyrido- [3,2-g]quinoline-3,7-dicarboxylate (9a) and diethyl 4,6-bis-(3-dimethyl- aminopropylamino) -10- methylpyrido[3,2-g]quinoline-3,7-dicarboxylate (9d) were able to inhibit cell proliferation of MCF-7 (Breast), NCI-H460 (Lung), and SF-268 (CNS) implying either amino or dimethylaminopropyl moiety at C-4 and C-6 positions is crucial for the antiproliferative activity of pyrido[3,2-g]quinoline derivatives. Compounds 9a – 9d were further evaluated for their activity against the growth of MCF-7 and two prostate cancer cell lines, LNCaP and PC-3. Results indicated the antiproliferative activity decreased in an order 9d > 9a >> 9b and 9c. Compound 9d was the most cytotoxic with an IC50 value of 5.63 and 3.96 ?戥M, respectively, against LNCaP and MCF-7. Flow cytometric analyses revealed that growth inhibition of LNCaP by 9d was due to cell cycle arrest in G1 phase, and followed by apoptosis. We further confirmed this 9d-induced apoptosis by Annexin V-FITC and cleavage pattern of PARP in LNCaP cells. To delineate the molecular mechanism of 9d-induced apoptosis, we examined the protein level of several cell cycle regulators including p53 and its downstream target(p21), cyclin D1, cyclin E, Rb, pRb and c-Myc. The results demonstrated that p53 was slightly increased but its target, p21, was not changed by treatment of 9d, suggesting p21 might not be a factor responsible for G1 arrest. Moreover, both cyclin D1 and cyclin E were decreased only in the presence of 80 ?戥M of 9d. In constrast, c-Myc, Rb and pRb were significantly reduced by 9d. Thus, we further examined the upstream regulators, Erk1/2 and p-Erk1/2 related to c-Myc. We found that 9d did not have the influence on Erk1/2, but it made p-Erk1/2 increase under the 5 ?戥M at 24 h. It indicated that the function of 9d to LNCaP cell cycle was probable by the effect of p-Erk1/2 on the decrease of c-Myc and then G1 arrest showed.
參考文獻
延伸閱讀
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