臺灣博碩士論文加值系統

Epirubicin是目前已被廣泛應用於抗腫瘤的藥物，其為細胞週期非特異性的抗腫瘤藥物，但使用於膀胱內灌注後所產生的治療效果並不 是很理想，一方面是因為部份藥物無法準確的傳送到腫瘤細胞，另一方面是由於灌注的藥物對膀胱會造成刺激性，且一般灌流的藥物停留在膀胱的時間只有2 個小時，又藥物對膀胱黏膜的滲透性不佳，因此限制了藥物滲入腫瘤組織。
Curcumin是生薑中的成分，近年來被廣泛研究及討論，文獻顯示其對於腫瘤的抑制也有顯著的療效，尤其是對於腫瘤細胞的G2/M 期。
因此本實驗的目的為將Epirubicin與Curcumin合併製成自體微乳劑，依Epirubicin和Curcumin不同的物化特性而將其分別均勻溶解於水相和油相中，期望此合併製劑可發揮加乘的治療效果。利用23複因子實驗設計法來探討界面活性劑的比例、輔界面活性劑的種類以及輔界面活性劑的比例對於粒子大小及包埋率的影響，從中選擇出粒徑及包埋率皆為最佳的處方進行後續物化特性的分析。實驗結果顯示出Epirubicin與Curcumin合併製成的自體微乳劑其粒子大小的測定值為150.7 ± 0.29 nm，且包埋率為98.31％，表面電荷的測定值為
-30mV。在體外溶離的實驗數據顯示出此劑型確實可增加Curcumin的釋出累積量達10.08倍及縮短Epirubicin HCl 釋出時間為原先的0.2倍。
進一步將此製劑進行T24人類膀胱癌細胞株的毒殺試驗，並以MTS asssay來評估細胞毒殺的效果，結果顯示Epirubicin與Curcumin合併製成的自體微乳劑相對於原本的Epirubicin HCl之細胞毒殺效果高出5.8倍，表示此合併製劑確實發揮了加乘的細胞毒殺作用，也的確為一值得繼續研究的抗膀胱癌製劑。

Epirubicin was the common drug used for antitumor therapy recently. It was a non-specific cell cycle antitumor drug, but the treatment outcome after intravesical therapy was not satisfied. Because of some parts of the drug were not transferred into cancer cell exactly, and the perfusion of drug irritated and limited the drug retention time inside bladder. The short retention time and poor drug permeability for bladder membrane limited anticancer treatment.
Curcumin was a discussed and investigated widely ingredient of ginger, many reports had shown that Curcumin has significant therapeutic effect for tumor inhibition, especially for G2/M phase of cell cycle.
The purpose of this study is to prepare Epirubicin and Curcumin loaded SMEDDS (self-microemulsifying drug delivery system), along with the different physical and chemical property of Epirubicin and Curcumin, we dissolved Epirubicin and Curcumin in aqueous phase and oil phase separately, expected this combine formulation can bring a synergistic antitumor activity. 2(3) factorial designs was used to estimate the effects of the surfactant adding ratio, co-surfactant type and co-surfactant adding amount on the particle size and the entrapment efficiency. To select the best formulation that had the smallest particle size and the highest entrapment efficiency for the follow up sequence analysis. The data shows that the mean particle size of the Epirubicin and Curcumin loaded microemulsion was 150.7 ± 0.29 nm, and the entrapment efficiency was 98.31 ％, zeta potential was -30mV. And the drug release experiments in vitro show that this formulation can increase the accumulated release amount and decrease the release time certainly.
The cytotoxic effects of Epirubicin and Curcumin loaded microemulsion, drug free microemulsion, Curcumin loaded microemulsion, Epirubicin loaded microemulsion and Epirubicin HCl on T24 bladder cancer cell lines were determined by MTS assay. The Epirubicin and Curcumin loaded microemulsion exhibiyed a higher cytotoxicity in T24 cell lines compared to the drug free microemulsion and Epirubicin HCl. In this study, the Epirubicin and Curcumin loaded SMEDDS showed a significant synergistic cytotoxicity, it may worth to explore for bladder cancer treatment.

目錄
表次目錄..................................................................................................IV
圖次目錄....................................................................................................V
中文摘要.................................................................................................VII
英文摘要..................................................................................................IX
壹、緒論.....................................................................................................1
一、前言.................................................................................................1
二、EPIRUBICIN HYDROCHLORIDE的基本概述......................................3
三、CURCUMIN的基本概述...................................................................5
四、高壓均質法簡介.............................................................................9
五、以實驗設計法探討.......................................................................10
六、膀胱癌簡介...................................................................................13
貳、實驗材料及儀器設備......................................................................18
一、實驗材料.......................................................................................18
二、儀器設備.......................................................................................19
三、培養液及緩衝液配置...................................................................21
參、研究方法及進行步驟......................................................................22
一、EPIRUBICIN之HPLC分析方法及檢量線的製作..........................22
(一) Epirubicin層析條件...................................................................22
(二) Epirubicin 檢量線之製作.........................................................23
(三) Epirubicin 定量方法之確效.....................................................23
二、CURCUMIN 之HPLC分析方法及檢量線的製作.........................24
(一) Curcumin 層析條件..................................................................24
(二) Curcumin 檢量線之製作..........................................................25
(三) Curcumin 定量方法之確效......................................................25
三、EPIRUBICIN自體微乳劑的製備.....................................................26
四、複因子試驗設計法.......................................................................28
五、EPIRUBICIN自體微乳劑之物化特性.............................................30
(一) 粒徑大小與表面電位測定分析...............................................30
(二) 型態觀察...................................................................................31
(三) 藥物含量評估...........................................................................31
(四) 示差掃描熱分析.......................................................................32
六、體外溶離試驗...............................................................................33
七、細胞毒殺作用...............................................................................34
肆、結果與討論......................................................................................38
一、藥品之定量分析...........................................................................38
(一) Epirubicin 的定量 .....................................................................38
(二) Curcumin 的定量......................................................................41
二、以實驗設計法探討處方...............................................................44
(一) 實驗設計法之探討...................................................................45
(二) 最佳化處方...............................................................................59
三、EPIRUBICIN自體微乳劑之物化性質評估.....................................60
(一) 粒徑大小分析...........................................................................60
(二) 表面電荷測定...........................................................................61
(三) 穿透式電子顯微鏡觀察...........................................................63
(四) 掃描式熱差分析儀評估...........................................................65
四、體外溶離試驗...............................................................................67
五、細胞毒殺作用之探討...................................................................69
伍、結論...................................................................................................81
陸、參考文獻..........................................................................................83

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