2012-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/647985摘要：白血病是血液的惡性腫瘤，主要是因血球造血細胞的生長不受控制、細胞的死亡減少、以及分化障礙。急性前骨髓性血癌(Acute promyelocytic leukemia, APL)是一種急性骨髓性血癌亞型，是血液及骨髓的一種癌症。因為這種急性症狀的進展很快以及其嚴重的複雜性病症，急性前骨髓性血癌被認為是急性白血病中，最嚴重的一種惡性腫瘤。Anthracyclines (例如daunorubicin) 是在1970到1980年間，最早被開發用來治療急性前骨髓性血癌病患。然而，醫學界對其治療的結果並不滿意，而其最主要的副作用—心臟毒性，則限制了這類藥物的使用。全反式維生素A 酸(All-trans retinoic acid)在急性前骨髓性血癌病患的治療，則是第一個以引發分化為作用機轉，來治療人類癌症的臨床藥物。全反式維生素A 酸的單一療法，或是與anthracyclines並用的合併療法，都顯示出不錯的臨床效果，然而，報導指出還是有約三分之一的病患會復發。三氧化二砷(Arsenic trioxide)則是另一種治療藥物，會引起急性前骨髓性血癌細胞的凋亡及部分分化作用。全反式維生素A 酸與三氧化二砷的合併療法，會加速癌細胞的萎縮及減少作用，主要就是經由增加細胞死亡及加速分化所導致。然而，雖然有不錯的臨床結果，三氧化二砷的安全性及耐受性則是最被擔憂的問題。皂苷則是一群天然的植物配醣體化合物，一般的分類上，皂苷可以分成三萜及固醇配醣體，或是分成三萜皂苷、spirostanol皂苷、furostanol皂苷。雖然皂苷曾被報導有免疫調節及抗癌作用(包括抗血癌作用)，但他們也有不好的作用，例如造成細胞膜穿孔。前期結果：在本計劃，我們將與藥物化學專家及臨床腫瘤專家合作(共同主持人)，我們將得到幾個系列的皂苷衍生物，並研究其作用。截至目前為止，共同主持人已合成多個衍生物，我們執行了一些前期試驗，評估其抗急性前骨髓性血癌細胞的作用。我們發現一個薯蕷皂苷衍生物具有很強的抗急性前骨髓性血癌細胞作用，但不會造成細胞溶破作用。經由蛋白陣列分析研究35種與凋亡相關的蛋白後，發現一些標的蛋白受到改變，包括HSP70及survivin的降解作用，以及一些蛋白的增加作用，如：促凋亡Bcl-2家族蛋白(Bim、Bax、Bad 及Bid)、死亡受體作用劑(FasL、TNF-α 及TNF-β)及死亡受體(Fas 、TNF-R1 、TNF-R2 、TRAILR-1/CD4 及TRAILR-2/CD5)，此外，此薯蕷皂苷衍生物也會促使CD40及CD40L的蛋白增加。特殊目的: 基於上述發現的結果，本研究計劃在第一年將詳細探討薯蕷皂苷衍生物，在抗急性前骨髓性血癌的確實作用機制及訊息傳遞，特別是受影響的蛋白、酵素、轉錄因子，接下來，我們將針對這些蛋白及因子，建構生物活性作用篩選系統，在這一年裡，除了已有的衍生物外，本計劃也會協助共同主持人合成更多衍生物，在本計劃第二年，我們將利用所建構的多種活性篩選系統，一一的來測試衍生物的活性，找出具開發潛力的衍生物。此外，也會執行抗急性前骨髓性血癌的合併療法，將活性衍生物合併現有的臨床治療藥物，如：anthracyclines、全反式維生素A 酸、三氧化二砷，合併療法的作用機轉也會深入研究。第三年則以動物實驗為主，除了建立異位異體腫瘤模式外(皮下打入癌細胞，雖然是血癌細胞，也是常用的腫瘤模式)，也將建立同位異體腫瘤模式(orthotopic xenograft tumor model)，執行內容除了衍生物的單一治療作用外，也會執行合併療法的動物實驗。我們期待經由本計劃所達成的成果，能發現更具活性及選擇性的衍生物，做更好的抗急性前骨髓性血癌的藥物研發。<br> Abstract: Leukaemia, the disease of haematological malignancies, is characterized by clonal expansion ofhematopoietic cells that are uncontrolled proliferation, decreased cell death as well as blockeddifferentiation. Acute promyelocytic leukemia (APL) is a subtype of acute myelogenous leukemia, acancer of the blood and bone marrow. Because of the rapid progression of the disease and severecomplications, APL was considered as the most malignant type of acute leukemia. Anthracyclines(e.g., daunorubicin) were firstly introduced for the treatment of APL patients in 1970 to 1980.However, the therapeutic outcome was not satisfactory. Cardiotoxicity, the major adverse effect,considerably limits the use of anthracyclines. All-trans retinoic acid for APL patients is the firsttreatment based on differentiation induction therapy of human cancer. Both the single use of all-transretinoic acid and the combinatory therapy of all-trans retinoic acid with anthracyclines reveal aconvincing clinical outcome. However, about one third of patients were reported to relapse. Arsenictrioxide induces apoptosis and partial maturation of APL cells. The combination of arsenic withall-trans retinoic acid accelerates tumor regression via increased differentiation and apoptotic celldeath. In spite of beneficial clinical outcome, there are still safety and tolerability issues for the use ofarsenic trioxide. Saponins are a group of naturally occurring plant glycosides. Normally, saponins aredivided into triterpenoid and steroid glycosides, or into triterpenoid, spirostanol, and furostanolsaponins. Although saponins may have immunomodulating and anticancer (including anti-leukemia)activities, they may also have untoward effects, such as membrane-permeabilizing effect.Preliminary data: In this project, we will collaborate with specialists in medicinal chemistry andclinical oncology, and will get several series of saponin derivatives. So far, we have obtained a lot ofderivatives and have determined their anti-APL activities. We found a diosgenyl saponin whichpotently induced APL cell death without the induction of cell lysis. Through the detection of theexpression of 35 apoptosis-related proteins, several signals have been identified, includingdown-regulation of HSP70 and survivin, and up-regulation of pro-apoptotic Bcl-2 family of proteins(Bim, Bax, Bad and Bid), death ligands (FasL, TNF-αand TNF-β)and death receptor (Fas, TNF-R1,TNF-R2, TRAILR-1/CD4 and TRAILR-2/CD5). Moreover, CD40 and CD40L were alsoup-regulated. Specific goals: Based on the positive results, there are several specific goals in theproject. In the first year, we will clearly identify the signaling cascades to the diosgenyl saponinaction and, in particular the targets and transcription factors involved in anti-APL activity. Next, wewill build compound examination templets based on the discovered molecular targets and willdetermine the activities of the derivatives on these targets in the second year. Besides, thecombination treatment of the potential derivatives with the clinical therapeutic drugs, includinganthracyclines (e.g., daunorubicin), all-trans retinoic acid and arsenic trioxide, will be examined inseveral APL cell lines. The signaling cascades of the combined therapy will be studied. In the thirdyear, a xenograft tumor model subcutaneously inoculated with APL cells and an orthotopic xenografttumor model of APL cells will be created and the single as well as combination treatment will betested in the models. We anticipate the achievement of these goals in the end of the project andsubject potential derivatives to advanced development.