2011-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/643811摘要：本計畫擬於三年內逐步完成幾項天然藥物活性成分之抗癌活性機轉探討。本實驗室將與美國北卡大學教堂山分校李國雄院士領導的天然藥物研究所-NPRL進行合作，目前將以QS-ZYX 和 DYZ衍生物作為主軸，本實驗室希望繼續致力於開發天然藥物作為抗癌藥物的來源。第一年的研究計畫中，我們將以DAPI stain和Cell Death ELISA kit等等，逐步證明QS-ZYX 和 DYZ衍生物能引起癌細胞凋亡，分析藥物作用濃度，來確認藥物的藥效，並回報資料，作為NPRL化學結構修飾及建立SAR的參考。此外，我們將針對這些QS-ZYX 和 DYZ衍生物的作用機轉做更進一步詳細的探討，運用xenograft動物模式做抗癌藥效評估及初步毒性觀察，也將委託泛球藥理研究所進行與癌症相關的kinases test測試，以期迅速找到藥物的主要作用點。臨床上癌症治療最大的障礙來自抗藥性的生成與癌細胞的轉移，本計畫第二年將觀察QS-ZYX 和 DYZ衍生物對於抗藥性蛋白p-glycoprotein與對癌細胞metastasis動物模式是否有抑制作用，並進一步探討其機轉，希望能增加其在臨床上應用的可能性。將委託泛球藥理研究所做廣泛性的enzyme screening，加速研究的廣度，並利用RT-PCR、Western blot等方式進一步確認。本研究計畫的第三年將利用數種動物模式，包括tumor xenograft、experimental metastasis assay等，來確認QS-ZYX 和 DYZ衍生物的抗癌及抑制癌轉移的藥效。由這些動物模式，我們可以觀察老鼠給藥後的生理病理狀態，做為是否需要修飾化合物結構，以增加藥物溶解度或是降低藥物毒性的參考。委託DCB及世宬，分別進行臨床前的毒理測試及PK實驗。藉由本研究計畫來篩選及分析出QS-ZYX 和 DYZ衍生物中的抗癌活性成分，探討其作用機轉，確認藥物在動物體內外作用情形，並進行臨床前試驗。希望本計畫的研究成果能促進相關的藥物開發並期望對相關的疾病治療有所貢獻。<br> Abstract: In this project we will cooperate with Prof. Kuo-Hsiung Lee at the University of North Carolina at Chapel Hill, School of Pharmacy, Natural Products Research Laboratories (NPRL), and QS-ZYX and DYZ series compounds will be the major project. In the former project, we found that the evodiamine, denbinobin, moscatilin and some other herbal constituents or synthetic compound (e.g. CIL-102) have anti-proliferation effect on several cancer cell lines. Therefore, we are interested in the development the natural products (including herbal medicines) as a new source of anti-cancer drug.There are three major studies in this project. In the first year, we will screen the modified derivatives of the active principles purified from natural products provided by NPRL. The apoptotic effects of these QS-ZYX and DYZ series compounds will be determined and quantified by DAPI stain, Cell Death ELISA kit, DNA ladder and flow cytometric analysis. We will obtain the inhibitory concentration ranges and the IC50 values, and then feedback these information to NPRL as a reference for chemical structure modification and build up the SAR. The action mechanism of active compounds selected from the QS-ZYX and DYZ series compounds will be studied. We also plan to use tumor xenograft model as an initial in vivo screening to provide the efficacy and toxicology data. This project also perform the cancer-related kinases screening (KinomeDiversityScreen™) by MDS Pharma Services, which could find out major targets of these active compounds.The major obstacles to clinical cancer therapy are multiple drug resistance and tumor metastasis. In the second year of this project, inhibitory effects of these natural product constituents on p-glycoprotein function and cancer invasion will be investigated. These action mechanisms and in vivo metastasis model will also be verified. The advantages of p-glycoprotein inhibition and anti-invasion benefit the clinical application of natural product constituents in chemotherapy. This project also perform the enzyme screening (SpectrumScreen®) by MDS Pharma Services, which could find out more targets of these active compounds. Early PK experiment will be tested by Rosetta Pharmamate Co., Ltd.After the in vitro experiments, we will focus the effective candidate compounds in in vivo studies, such as tumor xenograft model and experimental metastasis assay in the third year. Furthermore, several analyses will be performed in this work such as the histological analysis and staining, biochemical and molecular biological determinations, and pharmacological assessment. From these animal models, we can observe in vivo efficacy of the tested compounds and the compound-induced physiological and pathological change of mouse. These informations will be responded to NPRL to improve the solubility of the candidate compounds and reduce the adverse effects. Toxicology assay will be held by the Development Center for Biotechnology (DCB) and PK experiment by Rosetta Pharmamate Co., Ltd.Based on these assessments, the apoptotic and anti-invasive potential of QS-ZYX and DYZ series compounds will be elucidated and the potency will be confirmed in animal models. We hope that this project could contribute to the new drug development and clinical therapy.天然藥物細胞凋亡癌症轉移抗藥性Natural productsapoptosiscancermetastasisdrug resistanceThe Anti-Cancer Drug Development of Qs-Zyx and Dyz Series.