| 摘要: | 台灣每?有600 ?的?性腦瘤新病?產生並具有極高的致死?，患者的存活仍?過一至??，目 前?床所使用的生物可?解材質p(CPP-SA)結合BCNU 也僅能延長病患2 個月壽命。在本實驗室中， 長期致?於對人??性腦膠質瘤的新藥開發，近5 ?內目前已經發表?15 篇期刊?文，刊登於Clinical Cancer research、J Neurochem、Mol Pharmacol 等期刊，並取得美國、日本及中華民國五項專? (詳? 十二、計畫內容之(一) 近五?之研究計畫內容與主要研究成果?明)。而本實驗室和東華生物技術研 究所邱紫文教授已發展出生物可?解材質p(CPP-SA)的局部給藥系統，目前已經成功製作出 p(CPP-SA)，並將p(CPP-SA)攜帶本實驗室發現的抗癌物質n-butylidenephthalide，其體外釋放藥物情 形?好。動物實驗中也能有效抑制?性腦膠質瘤細胞的生長，此攜帶n-butylidenephthalide 的p(CPP-SA) 局部藥物釋放系統，目前正申請美國專?。 本計劃的共同主持人中國醫藥大學中國藥學研究所暨台灣大學化學系郭悅雄教授，從台灣特有的 台灣杉中純化出Taiwanin A，分子?350.32 kDa，化學名稱如下關鍵字中。Taiwanin A 和先前本實驗 室於南材胡純化出的ischaihulactone 其對於?性腦膠質瘤皆有顯著抑制其生長的效果，而Taiwanin A 其結構與isochaihulactone 十分相似，其結構上的只有?個位置具有差?，第一個是差?是在第七個 炭的位置上Taiwanin A 的是雙鍵，但是Isochaihulactone 是單鍵，第二個差?是在第?個炭的位置上 Taiwanin A 帶有的是methylenedioxy group，但是Isochaihulactone 帶有是3,4,5-trimethoxyphenyl group。 但是Taiwanin A 細胞毒?的效果較isochaiholactone 好上20~50 倍( Taiwanin A IC50 : 2~5 M vs. isochaiholactone IC50 : 100 M)。 本研究探討Taiwanin A 對於人??性腦膠質瘤的影響，發現其會使tubulin depolymerization，導 致細胞凋亡。我們發現Taiwanin A 對於?性腦膠質瘤的IC50 只有2~5 M，並且會造成Nonsteroidal Anti-inflammatory Drug-activated Gene (NAG-1)的表現上升，NAG-1 屬於TGF-，屬於一個抑癌基因。 由於在本實驗室先前研究已證實的NAG-1 表現的增加 (Chen, Lin et al. 2007) ，會誘導癌細胞的凋 亡，而histone acetylation 已證實會調控NAG-1 的表現 。由此讓我們?想到，Taiwanin A 造成細胞的 毒?與histone 的修飾有關。 因此本研究目標擬使用histone Modification Enzymes PCR Array、CHIP、microarray 方式探討是 否某些組織蛋白修飾的相關蛋白與Taiwanin A 造成?性腦膠質瘤細胞凋亡有關。?進一步，由於目 前使用於?床上以生物可?解p(CPP-SA)為基材的Glidel@ wafer，其攜帶藥物BCNU 具有較高毒性， 對於?性腦膠質瘤的抑制效果仍然有限，且其專?將於2010 ?過期，因此我們擬使用p(CPP-SA)攜 帶Taiwanin A 並以?鼠皮下攜帶?螢光人??性腦瘤動物模式和基因轉殖自發腦瘤小鼠動物模式， 評估以p(CPP-SA)攜帶Taiwanin A 治??性腦膠質瘤的可?性，以期提供腦瘤病患?有效的輔助治? 方式。
In Taiwan, there are 600 new cases of glioblastoma multiform (GBM). The lethal rate of GBM is very high and the lifespan of GBM patient is less than 2 years. In the clinical therapy, the biodegradable material p(CPP-SA) with BCNU could only prolong the 2 month lifespan. In our laboratory, we strive to develop new drugs for GBM therapy and published 15 papers in Clinical Cancer research 、Journal of Neurochemistry、Molecular Pharmacology in recent 5 years. We also get five patents in USA, Japan and Taiwan. We cooperate with professor Tzyy-Wen Chiou and have already developed the local release system of biodegradable material, p(CPP-SA). We use the p(CPP-SA) to carry the anticancer compound n-Butylidenephthalide and n-Butylidenephthalide could release from p(CPP-SA). In our in vitro and in vivo study, The n-butylidenephthalide-p(CPP-SA) can inhibit the growth of glioblastoma multiform effectively. We have applied for USA patents in the n-butylidenephthalide-p(CPP-SA) local release system. The other co-investigator, professor Yueh-Hsiung Kuo, in this plan extracted the pure compound, Taiwanin A, from Taiwania cryptomerioides, molecular weight 320.32 kDa. We found Taiwanin A could effectively inhibit the GBM cell growth as the same as the isochiahulactone, another pure compound extracted from Chaihu. The structure of Taiwanin A and isochaihulactone are similar and there are only two differences between their structures. First, in Tainwain A, it is double at C7 but it is single bond in isochiahulactone. Second, there is a methylenedioxy group at C6 in Taiwanin A but in isochaihulactone it is a 3,4,5-trimethoxyphenyl group. Interestingly, the cytotoxicity of Taiwanin A is greater 20 ~ 50 fold than isochaihulactone (Taiwanin A IC50 = 2~5 μM vs. Isochaihulactone IC50 = 100 μM). In this research, we find that Taiwanin A induce tubulin depolymerization and apoptosis in GBM cell. The Taiwanin A IC50 in GBM cells is 2~5 μM. We also found that the expression of Nonsteroidal Anti-inflammatory Drug-activated Gene (NAG-1) was induced by Taiwanin A in GBM cells. NAG-1 is a tumor suppress gene and belong to TGF-β family. In our previous study, it is demonstrated that the increased expression of NAG-1 induce apoptosis in cancer cells. Further, because it has been proved histone acetylation could regulate NAG-1 expression, it argue us that cytotoxicity of Taiwanin A is related with histone modification. Therefore, the aim of this study is to investigate if the cytotoxicity of Taiwanin A in GBM cells is related with histone modification by using histone Modification Enzymes PCR Array, CHIP and microarray. In addition, BCNU in the Glidel@ wafer, the clinical biodegradable BCNU-p(CPP-SA), is higher toxic to normal tissue. The inhibited ability of Glidel@ wafer on GBM cell is not very well. The patent of Glidel@ wafer will be overdue in 2010. In this study we would like to use the p(CPP-SA) to carry the Taiwanin A. The therapeutic effect of Taiwanin A-p(CPP-SA) will be evaluated by both theFGF1-SV40 transgenic mice model and the xenografts human GBM nude mice model. By this investigation, we hope to develop a more effective therapeutic method for brain tumor therapy. |
