2012-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/657229摘要：背景賀爾蒙療法抗藥性的前列腺癌是造成前列腺癌死亡的最主要因素，研究指出前列腺癌細胞的生長和生存經由細胞內的訊息傳導路徑所調控，因此發展抗癌藥物應用在前列腺癌之標靶治療成為我們的研究目標。本計劃擬定研發新的雜環分子篩選具有前列腺癌的抗癌活性，我們依據藥效基團以及分子模擬細胞內接受器和配體間相互作用的方式設計具有潛力的化合物，預計合成逾60 種化合物作為抗癌活性評估，而化合物由國際合作之俄國研究團隊設計並合成，建立化合物官能基之構效關係，未來將大量合成最具潛力之化合物並評估其動物實驗之生物活性。初步研究利用合成方法產出苯基吡咯烷（phenylpyrrolidine）衍生物和新的雜環分子衍生物，我們使用SRB 方法和CFSE 螢光染色法篩選具有抑制前列腺癌細胞增生的化合物。其活性化合物KUD773 可使細胞週期停滯在G2/M 期，並促使細胞進行計劃性死亡（細胞凋亡），利用西方點墨法發現KUD773 可使Cyclin B1 和MPM-2 蛋白表現量劇升，研究指出化合物具有抑制細胞進行有絲分裂的生物活性。目標第一年：合成三十個以上的化合物並進行抗癌活性測試，並且針對活性化合物探討其在前列腺癌細胞中的訊息傳導路徑，探究其生長因子的調控機制。第二年：以第一年的分子標靶作為合成化合物之模板，針對其活性化合物之衍生物進行分子生物抗癌活性的研究，預期其衍生物可作為標靶抑制劑成為有潛力的抗癌作用劑。第三年：大量合成活性化合物做體內活性測試，利用PC-3 和DU-145 異種移植腫瘤之動物實驗試驗，評估其化合物在動物體內之抗癌活性，並以動物模式探究其訊息傳導路徑，希望成為未來具有潛力的抗癌藥物。<br> Abstract: Background: Castration-resistant prostate cancer explains most of the prostatecancer-related deaths. Recent studies have identified the intracellular signaling pathwaysthat are critical to the growth and survival of the prostate cancer cells. The approachesworking on these intracellular targets may derive efficient therapeutic drugs against prostatecancers. The project will be directed on the development of novel heterocyclic entities withanticancer activity against prostate cancers. Design of potential active structures is based onknowledge of pharmacophoric fragments and molecular modeling of biologicaltarget–ligand interactions. More than sixty novel organic small-molecule compounds will besynthesized during the project and subsequently tested on antitumor activity. Syntheticmethods are based on the internal experience of the Russian research group.Structure–activity relationships will be determined for compounds under the study. Mostactive compounds will be resynthesized in large quantities for in vivo animal evaluation.Preliminary data: Several phenylpyrrolidine derivatives and novel heterocyclic derivativeshave been synthesized. The sulforhodamine B and CFSE staining assays demonstrated thatseveral derivatives displayed anti-proliferative activity against human hormone-refractoryprostate cancers. Furthermore, some of the active derivatives induced cell apoptosis whileKUD773, the most potent derivative, induced G2/M arrest of the cell cycle followed byprogrammed cell death of prostate cancer cells. Further identification showed that cyclin B1and MPM-2 were dramatically up-regulated by KUD773, indicating mitotic arrest to thecompound action. Specific goals: Based on the positive results, several specific goals willbe achieved in the three-year project. In the first year, more than thirty derivatives will besynthesized for advanced anticancer screening test. Furthermore, the anticancer signalingcascades induced by the effective derivatives will be studied and the critical intracellulartargets that regulate cell growth and survival will be discovered. In the second year, anotherthirty derivatives will be synthesized. Furthermore, the compound examination templetswill be set up based on the discovered molecular targets in the first year and all thederivatives will be examined in the established templets. We anticipate that several selectiveinhibitors acting on specific target will be discovered. In the third year, a large amount ofthe potential derivatives will be synthesized for the determination of in vivo efficacy. Thexenograft tumor models subcutaneously inoculated with PC-3 and DU-145 cells will becreated and the single as well as combination treatment will be tested in the models. Besides,the mechanism study associated with related signaling will be conducted in an in vivomodel.