本文選題：殼聚糖　切入點：5-氟尿嘧啶　出處：《廣東藥學院》2014年碩士論文　論文類型：學位論文

【摘要】：目的：化學合成5-氟尿嘧啶-殼聚糖-聚乙二醇單甲醚（5-FU-CS-mPEG）大分子前藥；制備5-FU-CS-mPEG納米粒；并考察其制備處方、體外釋放及動物藥效學行為。 方法：采用兩步合成法制備5-FU-CS-mPEG，首先在EDC/NHS的作用下，合成5-氟尿嘧啶-殼聚糖（5-FU-CS）前藥，再引入親水性mPEG，制備出親水性較好的5-FU-CS-mPEG前藥；采用紅外、1H-NMR及差示熱分析法表征合成產物。采用離子凝膠法制備5-FU-CS-mPEG納米粒，紫外分光光度法測定其載藥量，激光粒度分析儀測定粒徑、粒徑分布及Zeta電位，電子透射電鏡觀察其外觀形態(tài)，動態(tài)透析法考察其體外釋放情況并用各種釋放模型進行擬合。動物水平藥效學實驗，以昆明小鼠為實驗對象，建立H22肝癌移植瘤模型，考察高中低濃度5-FU-CS-mPEG納米粒、mPEG-CS納米粒及5-FU的抗癌活性及潛在的毒副作用，實驗結束后，計算其抑瘤率、脾指數(shù)及胸腺指數(shù)，并制腫瘤HE染色切片，實驗數(shù)據(jù)采用SPSS軟件處理、分析。 結果：經紅外、1H-NMR及差示熱分析法確證，已成功合成5-FU-CS-mPEG大分子前藥；通過對5-FU-CS合成工藝進行優(yōu)化，，得出制備5-FU-CS的最佳合成條件為EDC·HCl：5-FUA為1.5:1、EDC·HCl：NHS的為1:1、5-FUA：CS的摩爾比為1:1、反應時間為1d。紫外分光光度法測得5-FU-CS-mPEG納米粒的載藥量為4.2%，激光粒度儀測得空白納米粒（mPEG-CS）、載藥納米粒（5-FU-CS-mPEG）的平均粒徑分別為169.2nm、259.8nm，Zeta電位分別為+42.55mv、+39.27mv；透射電鏡觀察納米粒形態(tài)規(guī)則、分散性較好；體外釋放試驗發(fā)現(xiàn)，載藥納米粒在pH=5.5、pH=7.2的緩沖液中均有一定的緩釋作用， pH=5.5緩沖液中的釋放符合零級方程，pH=7.2緩沖液中的釋放符合Weibull模型。動物藥效學實驗顯示載藥納米粒高、中、低濃度組的抑瘤率分別為62.05%、48.79%及36.14%，與陰性對照組相比具有顯著性差異（P＜0.05），高劑量組抑瘤率與陽性對照組（5-FU）相當（P＞0.05）；病理組織切片顯示給藥組的細胞均出現(xiàn)不同程度地壞死、凋亡。 結論：采用離子凝膠法制備的載藥納米粒粒徑分布較均勻，形態(tài)規(guī)整；并且該載藥納米粒在體內外均表現(xiàn)較好的緩釋特性，動物實驗也表現(xiàn)出較強的抗腫瘤活性，因此該前藥納米制劑有望成為5-FU的新型載體。
[Abstract]:Aim: to synthesize 5-fluorouracil chitosan polyethylene glycol monomethyl ether 5-FU-CS-mPEG macromolecular prodrug, to prepare 5-FU-CS-mPEG nanoparticles, and to investigate the preparation of 5-FU-CS-mPEG nanoparticles. Methods: 5-FU-CS-mPEG was prepared by two-step synthesis. Firstly, 5-FU-CS-mPEG was synthesized by EDC/NHS, then hydrophilic mPEG was introduced to prepare 5-FU-CS-mPEG. 5-FU-CS-mPEG nanoparticles were prepared by ion-gel method, the drug loading was determined by ultraviolet spectrophotometry, the particle size, particle size distribution and Zeta potential were measured by laser particle size analyzer. Electron transmission electron microscopy (TEM) was used to observe its appearance, dynamic dialysis was used to investigate its release in vitro and all kinds of release models were fitted. The anticancer activity and potential side effects of 5-FU-CS-mPEG nanoparticles and 5-FU nanoparticles were investigated. After the experiment, tumor inhibition rate, spleen index and thymus index were calculated, and HE staining sections were made. The experimental data were processed by SPSS software. Analysis. Results: 5-FU-CS-mPEG prodrug was successfully synthesized by IR 1H-NMR and differential thermal analysis, and the synthesis process of 5-FU-CS was optimized. The optimum synthetic conditions for the preparation of 5-FU-CS were obtained as follows: the molar ratio of EDC 路HCl:5-FUA to 1.5: 1EDC 路HCl:NHS was 1: 1 and the reaction time was 1 day. The UV spectrophotometric method showed that the drug loading capacity of 5-FU-CS-mPEG nanoparticles was 4.2. The blank nanoparticles were obtained by laser particle size analyzer, and the drug-loaded nanoparticles 5-FU-CS-mPEG) were obtained. The average particle size is 169.2 nm ~ 259.8 nm ~ (-1) Zeta potential is 42.55 mv and 39.27 mv respectively, and the morphology of the nanoparticles is regular by transmission electron microscope (TEM). In vitro release test found that, The release of drug loaded nanoparticles in buffer solution of pH 5. 5 ~ 5 ~ (5) ~ (5) ~ (2) ~ (2) ~ (2) was consistent with the Weibull model. The results of animal pharmacodynamics experiments showed that the drug loaded nanoparticles were high and medium, and the release in pH = 5. 5 buffer solution was in accordance with the zero order equation (pH = 5. 5) and the release in pH ~ (2 +) buffer was consistent with the Weibull model. The tumor inhibition rates in the low concentration group were 48.79% and 36.14%, respectively, which were significantly different from those in the negative control group (P < 0.05). The tumor inhibition rate in the high dose group was similar to that in the positive control group (P > 0.05). Conclusion: the drug loaded nanoparticles prepared by ion-gel method have more uniform particle size distribution and regular morphology, and the drug loaded nanoparticles exhibit good sustained release characteristics in vivo and in vitro, and the animal experiments also show strong anti-tumor activity. Therefore, the prodrug nanometer preparation is expected to become a new carrier of 5-FU.
【學位授予單位】：廣東藥學院
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R943;R965

【參考文獻】

相關期刊論文 前10條

1 李東旭;耿燕麗;;殼聚糖的改性及作為生物材料的應用研究[J];材料科學與工程學報;2006年02期

2 徐文峰;廖曉玲;;堿量法測定殼聚糖脫乙酰度的研究[J];分析試驗室;2008年S1期

3 張建新;蔡遜;金煒東;張智勇;王向昱;;三氧化二砷聯(lián)合5-氟尿嘧啶對小鼠肝癌H_(22)細胞移植瘤的作用[J];肝膽胰外科雜志;2012年02期

4 龐國棉;朱亮;李遠達;;5-氟尿嘧啶的聚乙二醇-聚乳酸的膠束制備及其體外釋藥研究[J];廣東藥學院學報;2011年02期

5 李愛貴,鄧聯(lián)東,董岸杰;聚乙二醇在新型藥物制劑中的應用[J];高分子通報;2004年04期

6 黃攀;韓寶芹;劉萬順;常菁;董文;;N-羧甲基殼聚糖的制備及其生物相容性評價[J];功能材料;2009年07期

7 陳建勇,劉冠峰;殼聚糖改性及用其整理紡織品抗菌性能的研究[J];功能高分子學報;2002年02期

8 王愛勤,俞賢達;烷基化殼聚糖衍生物的制備與性能研究[J];功能高分子學報;1998年01期

9 姚金鳳;郭晨陽;楊紅;譚桂蓮;薛明;;殼聚糖納米粒在藥物輸送中的應用研究進展[J];國際藥學研究雜志;2013年01期

10 段萍萍;關鵬程;朱亮;;聚乙二醇化殼聚糖-氟尿嘧啶偶合物的制備及體外釋放研究[J];廣東藥學院學報;2013年06期

相關博士學位論文 前2條

1 陸曉;陽離子聚合物PC偶聯(lián)小分子藥物協(xié)同基因的抗腫瘤活性研究[D];浙江大學;2011年

2 熊靜;5-氟尿嘧啶氨基酸類化合物的合成及抗癌活性[D];大連理工大學;2009年

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