臺灣博碩士論文加值系統

脂溶性藥物-薑黃素有溶解度低、穩定性差和口服生物可利用率低的缺點，利用微乳液的劑型配合經皮輸藥系統可以克服上述缺點，提升藥物治療效果。利用經皮輸藥系統的途徑，最大的阻礙為皮膚的角質層蔽障，要克服皮膚的角質層蔽障，目前已經有物理和化學的方式來增加藥物的穿透量。本研究利用化學方式篩選多種促進劑，結果顯示發現油相limonene、geraniol、Isopropyl myristate (IPM)、醇類isopropanol (IPA)和陽離子界面活性劑、Dimethyl dioctadecyl ammonium bromide (DDAB)、N-cetyl-N,N,N,trimethyl amoniumbromid (CTAB)具有好的經皮吸收促進效果，利用此配方搭配6 % tween 80界面活性劑震盪可以得到約21.59 nm 之Microemulsion.A (ME.A)和limonene、geraniol、Isopropyl myristate搭配6 % Tween 80 可以得到93.32 nm之Microemulsion.E (ME.E)。利用體外穿透裝置24 hr薑黃素藥物總穿透量Microemulsion.A (ME.A)可以達98 μg/cm2，Microemulsion.E (ME.E)可以達82 μg/cm2，示差掃描量熱儀 (DCS)也證實微乳液是經由細胞間質脂質穿過皮膚。當自我乳化微乳液配方確定後，則進行細胞刺激性、溶血實驗和皮膚組織切片及儲存安定性實驗之探討。在油酸微乳液配方D (OA.D)上，利用體外穿透裝置24 hr薑黃素藥物總穿透量達27μg/cm2 ，實驗之結果證明開發之微乳液具有一定安定性且無毒性，有效提高脂溶性藥物溶解度並且增加經皮穿透效率。

The lipophilic drug, curcumin, has low oral bioavailability because of its low solubility and poor stability. By using microemulsion via the transdermal route, a new delivery system was developed to improve curcumin’s delivery. Stratum corneum in skin is the main obstacle in transdermal drug delivery systems. To overcome the stratum corneum barrier, several physical and chemical means to increase the drug penetration have been studied. In this study, a variety of chemical enhancers were screened with the help of Plackett Burman design to evaluate their effects on curcumin transderaml delivery. We found that limonene, geraniol, isopropyl myristate (IPM), isopropanol (IPA), and Dimethyl dioctadecyl ammonium bromide (DDAB)、N-cetyl-N,N,N, trimethyl amoniumbromid (CTAB) had good enhance effects on the percutaneous absorption of curcumin.
Microemulsion.A (ME.A) containing limonene, geraniol, isopropyl myristate isopropanol, DDAB, CTAB, and 6 % tween 80 has a nano-scale particle size (21 nm) and a transparent appearance. Microemulsion.E (ME.E) containing limonene, geraniol, isopropyl myristate, and 6 % tween 80 has a nano-scale particle size (93 nm). By using the in vitro Franz diffusion cells, curcumin loaded Microemulsion.A (ME.A) can penetrate the pig ear skin at a flux up to 98 μg/cm2 in 24 hrs. Microemulsion.E (ME.E) can reach a 82 μg/cm2 in 24 hrs. The DSC thermogram result showed that the intracellular transport is possible mechanism of ME.A and ME.E. We also used cytoxicity irritation test and hemolysis of red blood cells to evaluate the microemulsion(ME) toxicity. These results showed that the developed ME was non-toxicity and stable. However, Oleic acid microemulsion D (OA.D) had a transdermal delivery rate of 27 μg/cm2 in 24 hrs. In our transdermal study. This new delivery system has the potential to be applied to other lipophilic durgs to enhance their solubility and delivery efficiency.

摘　要 i
Abstract vi
目　錄 viii
圖目錄 xi
表目錄 xiv
第一章 研究動機 1
第二章 緒論 3
2-1 皮膚介紹及經皮輸藥 3
2-1-1 皮膚結構 3
2-1-2 藥物穿透皮膚途徑 5
2-1-3 藥物於皮膚之作用部位 6
2-1-4皮膚製劑的改善方式 7
2-1-5 促進劑增加經皮吸收之數學模式 13
2-1-6 經皮藥物傳遞的優點和限制 17
2-2 薑黃素 18
2-2-1 薑黃素簡介 18
2-2-2 薑黃素的物理化學性質 19
2-2-3 薑黃素於臨床上的研究 19
2-2-4 薑黃素所面臨個問題 22
2-3 劑型介紹 25
2-3-1微乳液簡介 25
2-3-2微乳液之熱力學穩定性 25
2-3-3 微乳液型態 26
2-3-4 微乳液應用於經皮輸藥優點 28
第三章 材料和儀器設備 29
3-1 材料 29
3-2 儀器設備 30
第四章 實驗方法 31
4-1 實驗動物皮膚製備 31
4-2 體外穿透裝置及試驗方式 31
4-3 豬皮薑黃素濃度測量 32
4-4油酸微乳液擬三項圖繪製 33
4-5 油酸微乳液製作方式 33
4-6 自我乳化微乳液擬三項圖繪製 34
4-7 自我乳化微乳液製作方式 35
4-8 高效液相層析儀分析薑黃素含量 35
4-8-1 薑黃素於HPLC分析條件 35
4-8-2 薑黃素檢量線製作 36
4-9 數據統計及分析 36
4-10 細胞刺激性實驗 37
4-11 RBC試驗 37
4-11-1製備紅血球混合溶液 37
4-11-2溶血實驗 37
4-12 粒徑大小及粒徑分佈指數測量 38
4-13 表面電位測定 38
4-14 共軛焦顯微鏡分析條件 39
4-15 穿透式電子顯微鏡分析條件 39
4-16示差掃描量熱儀(DSC)分析條件 40
4-17皮膚組織石蠟包埋染色切片 40
4-18 促進劑篩選 40
第五章 結果與討論 44
5-1 油酸微乳液劑型 44
5-1-1 油酸微乳液三相圖 45
5-1-2 油酸微乳液經皮穿透 46
5-1-3 油酸微乳液細胞刺激性 48
5-2 化學促進劑篩選 49
5-3 化學促進劑第二次篩選 53
5-4自我乳化微乳液劑型 56
5-4-1 ME.1-ME.5物化性質 56
5-4-2 ME.A-ME.H物化性質 57
5-4-3 ME.A三相圖 60
5-4-4 ME.A、ME.E穿透式電子顯微鏡觀察微乳液型態 61
5-4-5 ME.A ME.E載體安定性實驗 62
5-4-6 不同比例tween 80 微乳液經皮穿透 64
5-4-7 不同配方之微乳液經皮穿透 66
5-4-8 ME.A ME.E半透膜藥物釋放模式 72
5-4-9 ME.A ME.E於豬皮藥物釋放模式 73
5-4-10 ME.A包覆不同薑黃素含量穿透試驗 75
5-4-11 ME.A、ME.E細胞刺激性實驗 77
5-4-12 ME.A、ME.E溶血率實驗 78
5-4-13共軛焦顯微鏡分析ME.A、ME.E載體皮內藥物分佈 79
5-4-14 DSC觀察ME.A、ME.E載體藥物穿透形式 84
5-4-15 H&E豬皮組織染色圖 86
結論 87
參考資料 89

 
圖目錄
圖 2-1 人類皮膚縱剖圖 [5] 4
圖 2-2 藥物穿透角質層模式[9] 6
圖 2-3 藥物穿透的疏水和親水路徑及促進劑作用模式[9] 7
圖 2-4 脂肪酸結構[15] 8
圖 2-5 常見萜烯類結構[17] 11
圖 2-6穩定狀態下藥物穿透生物屏障之擴散圖 13
圖 2-7 薑黃素的分離和萃取過程[22] 18
圖 2-8 薑黃素及其天然衍生物結構[23] 19
圖 2-9 薑黃素多重治療效果[21] 20
圖 2-10 薑黃素和其延伸物及代謝物[21] 23
圖 2-11 微乳液擬三相圖 [38] 27
圖 4-1 Franz diffusion cell，(a) 示意圖 (b) 實際裝置圖[44] 32
圖 4-2 薑黃素之檢量線 36
圖 5-1 油酸微乳液三相圖 45
圖 5-2 OA.1至OA.6以不同比例之油酸微乳液之穿透豬耳外側皮膚24小時累積穿透量(n=2) 。 46
圖 5-3 OA.A-OA.G以不同oleic acid及Tween 80 / ETOH比例之微乳液穿透豬耳外側皮膚24小時累積穿透量(n=2) 47
圖 5-4 OA.D細胞刺激性實驗(n=3) 48
圖 5-5 24種不同化學促進劑於 PBS/ETOH=1:1，穿透豬耳外側皮膚24小時之累積穿透量(n=2) 50
圖 5-6 24種不同化學促進劑於PBS/ETOH = 1:1 ，穿透豬耳外側皮膚24小時之累積穿透量(n=2) 50
圖 5-7 篩選過之化學促進劑於PBS/ETOH = 1:1 ，穿透豬耳外側皮膚24小時之累積穿透量(n=2) 53
圖 5-8 無包覆薑黃素和有包覆薑黃素之ME.A 58
圖 5-9 ME.A三相圖 60
圖 5-10 TEM下ME.A外觀型態 61
圖 5-11 TEM下ME.E外觀型態 61
圖 5-12 ME.A、M.E.E粒徑大小隨時間變化(n=3) 63
圖 5-13 ME.A、ME.E薑黃素量隨時間變化(n=3) 63
圖 5-14 Control、ME.1-ME.4分別為0 %、2%、4%、6%、12 % Tween 80比例之乳液穿透豬耳外側皮膚24小時累積穿透量(n=3) 65
圖 5-15 Control、ME.1-ME.4分別為0 %、2%、4%、6%、12 % Tween 80比例之乳液薑黃素模式藥物殘留於豬皮24小時累積穿透量(n=3) 65
圖 5-16 微乳液A至E，豬耳外側皮膚24小時穿透曲線(n=3) 67
圖 5-17 微乳液A至E配方比較，乳液殘留於豬皮24小時累積穿透量(n=3) 68
圖 5-18 微乳液A和F-H配方，豬耳外側皮膚24小時穿透曲線(n=3) 70
圖 5-19 微乳液A、E、F、G、H配方比較，乳液殘留於豬皮24小時累積穿透量(n=3) 71
圖 5-20 ME.A、ME.E及控制組藥物釋放(n=3) 72
圖 5-21 ME.A於豬皮之藥物釋放模式 74
圖 5-22 ME.E於豬皮之藥物釋放模式 74
圖 5-23不同薑黃素量之ME.A(n=3) 75
圖 5-24不同薑黃素量之ME.A穿透總量和濃度關係(n=3) 76
圖 5-25 ME.A、ME.E細胞刺激性實驗(n=3) 77
圖 5-26 ME.A、ME.E對紅血球溶血實驗(n=3) 78
圖 5-27 ME.A包覆薑黃素皮內螢光強度和深度關係 80
圖 5-28 A : ME.A薑黃素溶液藥物滲透於豬皮明視野B : ME.A薑黃素溶液藥物滲透於豬皮之疊圖並平均螢光。 80
圖 5-29 ME.E薑黃素溶液皮內螢光強度和深度關係 81
圖 5-30 A : ME.E薑黃素溶液藥物滲透於豬皮明視野，B : ME.E薑黃素溶液藥物滲透於豬皮之疊圖並平均螢光 81
圖 5-31 Control薑黃素溶液皮內螢光強度和深度關係 82
圖 5-32 A : Control薑黃素溶液滲透於豬皮明視野，B : Control薑黃素溶液滲透於豬皮疊圖並平均螢光。 82
圖 5-33不同配方皮內螢光強度與深度比較(n=1) 83
圖 5-34 皮膚於DSC圖譜。溫度設定30-90℃ 85
圖 5-35皮膚於DSC圖譜。溫度設定30-140℃ 85
圖 5-36 豬皮H&E組織染色切片(20X) 86


 
表目錄
表 2-1 萜烯類分類[17] 9
表 2-2 不同經皮促進方式比較[18] 13
表 2-3 目前進行中之薑黃素於各種疾病治療之各期臨床研究[22] 21
表 2-4 薑黃素的生體利用率[24] 23
表 2-5微乳液與乳液比較[37] 26
表 4-1 不同比例之油酸微乳液 33
表 4-2 不同oleic acid及tween 80/ETOH比例之微乳液組成 34
表 4-3 不同比例Tween 80之微乳液 35
表 4-4 不同促進劑之微乳液 35
表 4-5 Plackett Burman design不同化學促進劑篩選 41
表 4-6 Plackett Burman design化學促進劑篩選 42
表 4-7 Two Level factorial design 化學促進劑篩選 43
表 5-1 OA.1至OA.6以不同比例之油酸微乳液，其經皮穿透參數(n=2) 46
表 5-2 OA.A-OA.G以不同oleic acid及Tween 80 / ETOH比例，其經皮穿透參數(n=2) 47
表 5-3 不同化學促進劑篩選，其經皮穿透參數、薑黃素穿透總量和豬皮殘留量(n=2) 51
表 5-4 化學促進劑之經皮穿透迴歸分析 52
表 5-5 化學促進劑篩選A至P，其經皮穿透參數、薑黃素穿透總量和豬皮殘留量(n=2) 54
表 5-6 化學促進劑之經皮穿透迴歸分析 55
表 5-7 包覆薑黃素之ME.1至ME.5粒徑大小和粒徑分散指數和介面電位(n=3) 56
表 5-8 包覆薑黃素之ME.1至ME.5之物理性質(n=1) 57
表 5-9 包覆薑黃素之ME.A至ME.E粒徑大小和粒徑分散指數和介面電位(n=3) 58
表 5-10 包覆薑黃素之ME.A至ME.E之物理性質(n=1) 59
表 5-11微乳液A至E，其經皮穿透參數(n=3) 67
表 5-12微乳液A和F-H，其經皮穿透參數(n=3) 70
表 5-13不同薑黃素之ME.A，其經皮穿透參數(n=3) 76

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