臺灣博碩士論文加值系統

Tiopronin (N-(2-Mercaptopropionyl)-glycine) 為一具有保護肝臟、代謝系統的解毒作用之mercaptopropionic acid衍生物。除此之外，Tiopronin亦具有增加重金屬排泄、降低cystine的濃度以及預防白內障之作用。
本研究以牛血清白蛋白為基礎，運用乳化-熱穩定技術 (emulsification–heat stabilization technique) 形成包覆Tiopronin之白蛋白微球 (microsphere)，再以不同比例聚合物和藥物所製備之微球來探討其性質的相互關係。利用高效能液相層析儀檢測微球所包覆的藥物含量；使用光學顯微鏡觀察微球的粒徑分佈及表面性質，並測試其在不同條件下之溶出行為，以及體內試驗中肝臟修復程度之觀察。實驗結果顯示所製備的微球具有光滑的表面，而提高聚合物比例可提高藥物包覆率；同時，在體外釋出試驗發現，Tiopronin之初始釋出速率隨聚合物之比例之增加而下降，但改變聚合物比例與微球粒徑大小並無相對關係；此外，體內試驗結果顯示，Tiopronin白蛋白微球並未出現靶向 (drug target) 效果；另一方面，於試驗開始後第六週，出現明顯之藥理效果，其效果與Silymarin相似。

Tiopronin (N-(2-Mercaptopropionyl)-glycine) is a derivative of mercaptopropionic acid known to protect the liver, the detoxification of metabolic system. Also, it can enhance the excretion of heavy metals, lower cysteine concentrations and prevent cataract.
In this study, we utilized bovine serum albumin as base to incorporate Tiopronin by using emulsification–heat stabilization technique. The prepared albumin microsphere, with different proportions of the polymers and drugs, were used to explore the relationship between composition of microsphere and properties. Tiopronin loading in microsphere was measured by using high-performance liquid chromatography (HPLC) method. This study employed an optical microscope to observe the particle size distribution and surface properties of microsphere. Furthermore, this study tested the releasing behaviors in different conditions and the degree of liver repair observed in-vivo test. The result shown the formulated microsphere has smooth surface, and the drug loading efficiency is enhanced by increasing the ratio of polymer in formulation. Also, in-vitro test showing increases of the initial releasing rate of Tiopronin by reducing the ratio of polymer in the formulation, but the relative relationship between polymer ratio and particle size of microspheres was not observed. Additionally, the in-vivo test result of Tiopronin albumin microparticles showed no drug target effect. The sixth week after the treatment, it''s shown apparent pharmacological effect, and the effect is similar to Silymarin.

中文摘要--------------------------------------------------------------------------- I
英文摘要-------------------------------------------------------------------------- II
誌謝------------------------------------------------------------------------------- III
目錄------------------------------------------------------------------------------- IV
表次目錄----------------------------------------------------------------------- VIII
圖次目錄------------------------------------------------------------------------- IX


本文
壹、前言-------------------------------------------------------------------------- 1
貳、文獻回顧-------------------------------------------------------------------- 3
一、Tiopronin (活性藥物成分) 之基本概述---------------------------- 3
(一) 物化性質--------------------------------------------------------- 3
(二) 藥理性質--------------------------------------------------------- 4
(三) 藥動性質--------------------------------------------------------- 5
(四) 適應症/用法用量----------------------------------------------- 6
(五) 副作用------------------------------------------------------------ 6
¬二、血清白蛋白 (Serum albumin) --------------------------------------- 7
三、微球製備技術 (Microsphere formation) --------------------------- 8
(一) 微球之特性------------------------------------------------------ 8
(二) 微球製備方法--------------------------------------------------- 9
四、肝臟介紹---------------------------------------------------------------- 10
(一) 肝臟簡介-------------------------------------------------------- 10
(二) 肝功能指數----------------------------------------------------- 10
(三) 肝纖維化與肝硬化簡介-------------------------------------- 11
五、肝損傷之動物實驗模式---------------------------------------------- 12
參、實驗材料與儀器設備---------------------------------------------------- 14
一、實驗材料（Materials) ------------------------------------------------ 14
(一) 藥物-------------------------------------------------------------- 14
(二) 化學試藥-------------------------------------------------------- 14
(三) 鼠體品系-------------------------------------------------------- 15
(四) 動物試驗器具及耗材----------------------------------------- 15
二、儀器設備 (Apparatus) ----------------------------------------------- 16
肆、實驗方法------------------------------------------------------------------- 17
一、Tiopronin之定量分析方法------------------------------------------ 17
(一) 溶液配製-------------------------------------------------------- 17
(二) 高效能液相層析儀 (HPLC) 分析方法--------------------- 17
二、Tiopronin定量分析方法之分析方法確效------------------------ 18
(一) 系統適用性 (System suitability) -------------------------- 18
(二) 專一性 (Specificity) ----------------------------------------- 19
(三) 線性 (Linearity) 與範圍 (Range) ------------------------ 20
(四) 精密度 (Precision) ------------------------------------------- 20
(五) 準確度 (Precision) ------------------------------------------- 22
三、微球劑型製備---------------------------------------------------------- 23
(一) 微球製備處方-------------------------------------------------- 23
(二) 製程操作-------------------------------------------------------- 24
四、製劑中的tiopronin含量分析--------------------------------------- 25
(一) 標準溶液之製備----------------------------------------------- 25
(二) 檢品溶液之製備----------------------------------------------- 25
(三) 製劑中tiopronin之定量分析------------------------------ 25
五、微球型態觀察及粒徑大小測定------------------------------------- 26
六、示差掃描熱分析儀---------------------------------------------------- 26
(一) 儀器設定-------------------------------------------------------- 26
(二) 實驗步驟-------------------------------------------------------- 26
七、體外釋出試驗---------------------------------------------------------- 27
(一) Medium---------------------------------------------------------- 27
(二) 試驗方法-------------------------------------------------------- 27
八、動物試驗 (In vivo) --------------------------------------------------- 28
(一) 實驗設計-------------------------------------------------------- 28
(二) 實驗步驟-------------------------------------------------------- 29
(三) 組織切片及染色過程----------------------------------------- 30
(四) 組織切片病理觀察-------------------------------------------- 31
(五) 統計方法-------------------------------------------------------- 32
伍、結果與討論---------------------------------------------------------------- 33
一、Tiopronin定量分析方法之分析方法確效------------------------ 33
(一) 系統適用性 (System suitability) -------------------------- 33
(二) 專一性 (Specificity) ----------------------------------------- 35
(三) 線性 (Linearity) 與範圍 (Range) ------------------------ 36
(四) 精密度 (Precision) ------------------------------------------- 36
(五) 準確度 (Precision) ------------------------------------------- 38
二、微球劑型製備---------------------------------------------------------- 39
三、微球包覆率及型態觀察---------------------------------------------- 40
(一) PM系列處方---------------------------------------------------- 40
(二) BM系列處方---------------------------------------------------- 40
(三) 處方包覆率比較----------------------------------------------- 41
四、微球粒徑大小測定---------------------------------------------------- 41
五、Tiopronin微球製劑之DSC------------------------------------------ 43
六、體外釋出試驗---------------------------------------------------------- 44
七、動物試驗 (In vivo) --------------------------------------------------- 48
(一) 大鼠體重變化-------------------------------------------------- 48
(二) 動物試驗之外在觀察----------------------------------------- 48
(三) 血清中GOT及GPT數值之變化--------------------------- 49
(四) 血清中triglyceride及cholesterol數值之變化------------ 50
(五) 組織病理學之觀察------------------------------------------- 57
陸、結論------------------------------------------------------------------------- 66
柒、參考文獻------------------------------------------------------------------- 67
捌、附錄------------------------------------------------------------------------- 73

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