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中文摘要： 本文的实验目的：（1）考察影响非靶向性聚乳酸（PLA）载药纳米粒制备的工艺因素，并对处方进行工艺优化，制备出粒径较小、载药率和包封率较高的PLA载药纳米粒；（2）探索不同反应条件对肺靶向配体分子叶酸（FA）和透明质酸（HA）偶联量的影响，确定配体分子的最佳偶联工艺；（3）通过模拟体内环境考察不同因素对纳米粒释药性能的影响，分析释药机理并建立合适的释药模型，为药物在体内的药效学研究奠定基础。采用的实验方法：（1）采用复乳法（W/O/W）和乳化溶剂挥发法（O/W）分别制备奥沙利铂PLA纳米粒、吉西他滨PLA纳米粒和紫杉醇PLA纳米粒，对两种制备方法中影响纳米粒粒径和载药率的因素进行单因素考察，筛选出较好的水平设计正交实验，优化得到最佳制备工艺。通过透射电子显微镜（TEM）和Zeta电位及激光粒度分析仪对纳米粒形貌和粒径进行表征；（2）过酰胺化反应分别将FA和HA偶联到PLA载药纳米粒表面，考察不同反应条件对配体偶联量的影响并找出最佳偶联工艺。红外光谱（IR）表征配体偶联情况，Zeta电位及激光粒度分析仪和扫描电子显微镜（SEM）对偶联配体后的纳米粒粒径和形貌进行表征；（3）考察不同PH值（PH=4.3、PH=5.6和PH=7.4）、不同载药率（9.53%、17.48%和26.37%）以及偶联配体对纳米粒释药性能的影响，并模拟人体内部环境进行释药行为的研究，分析每个阶段的释药机理，拟合五种常见药物模型的曲线方程并选择最佳释药模型。实验结果：（1）通过单因素考察和正交试验设计确定了制备三种载药纳米粒的最佳制备工艺，在最佳制备条件下制得的载药纳米粒载药率和包封率分别为26.9%，66.5%；27.4%，67.2%和30.3%，73.6%。纳米粒表面光滑圆整、分散性好，平均粒径分别为110nm、117nm和125nm，且粒径分布范围较窄；（2）用红外光谱（IR）表征纳米粒表面分子基团的变化，表明FA和HA分别通过酰胺键作用成功的偶联到了纳米粒表面。确定了FA和HA的最佳偶联工艺，在最佳偶联反应条件下FA和HA的偶联量分别为0.0237×10-3mol/g和0.0178×10-6mol/g。粒径分析表明FA偶联前后纳米粒粒径基本无变化，HA（Mr=300000）偶联后粒径由122.2nm增长到131.3nm，表明大分子量的HA对粒径变化影响较大。通过放大不同倍数的SEM照片可看出配体偶联后纳米粒分布均匀，分散性较好；（3）纳米粒释药速率和累积释药量顺序为PH=4.3＞PH=5.6＞PH=7.4，且在一定PH范围内酸性越强释药速率和累计释放量越高，高载药率纳米粒在相同环境下的释药速率高于中、低载药率纳米粒，FA偶联后对纳米粒释药性能基本无影响，大分子的HA偶联到纳米粒表面后释药速率明显降低。靶向PLA载药纳米粒模拟体内环境释药表现出良好的缓释效果，拟合的FA/HA-PLA载药纳米粒的释药模型分别为InIn(1/(1-Q))=0.04633Int-1.03562和1-Q1/3=-0.01368t+1.04726。 google翻译： The purpose of this experiment: (1) examine the impact on non-target polylactic acid (PLA) drug-loaded nanoparticles preparation of process factors, and prescription process optimization, the preparation of a smaller particle size, drug loading and encapsulation efficiency. high PLA drug-loaded nanoparticles; (2) to explore different reaction conditions for lung targeting ligand folic acid (FA) and the amount of hyaluronic acid (HA) coupling to determine the optimal coupling of the ligand molecule process; (3) examine the impact of different factors on the drug release properties of nanoparticles by simulating the in vivo environment, analysis of the release mechanism and the establishment of appropriate release model, to lay the foundation for the drug in vivo pharmacodynamic studies. Experimental methods: (1) double emulsion (W / O / W) emulsion solvent evaporation method (O / W) Preparation of of oxaliplatin PLA nanoparticles, the gemcitabine PLA nanoparticles and paclitaxel PLA nanoparticles, two preparation methods of nanoparticles particle size and drug loading rate single factor study, filter out the good level orthogonal experiments to optimize the best preparation process. By transmission electron microscopy (TEM) and Zeta potential and laser particle size analyzer nanoparticle morphology and particle size characterization; (2) Over amidation reaction of the FA and HA coupling to the surface of the PLA drug-loaded nanoparticles investigated The reaction conditions on the amount of ligand coupled and find out the best coupling process. Infrared spectroscopy (IR) characterization of ligand coupling, zeta potential and laser particle size analyzer and scanning electron microscope (SEM) coupled ligand nanoparticle size and morphology were characterized; (3) study at different pH values ( pH = 4.3 and pH = 5.6 and pH = 7.4), different drug loading rate (9.53%, 17.48% and 26.37%, respectively), and the of coupling ligands on the drug release properties of nanoparticles and simulate the body's internal environment release behavior research, analysis of each stage of the release mechanism, fitting the curve equation model of five common drugs and select the best drug delivery model. Results: (1) single factor and orthogonal design to determine optimal preparation prepared three kinds of drug-loaded nanoparticles, drug loading and encapsulation efficiency obtained in the best preparation conditions of drug-loaded nanoparticles were 26.9%, 66.5%; 27.4%, 67.2% and 30.3%, 73.6%, respectively. Nanoparticle surface is smooth and round, good dispersion, the average particle size of 110nm, 117nm and 125 nm, and the particle size distribution is narrow; (2) changes in the nanoparticle surface molecular groups characterized by infrared spectroscopy (IR), indicating that the HA and FA were successful through the amide key role of coupling to the surface of the nanoparticles. The FA and HA coupling process, under the reaction conditions in the top coupling a coupling amount of FA and HA were 0.0237 × 10-3mol / g and 0.0178 × 10 6 mol / g. The particle size analysis showed that the nanoparticles before and after the FA coupled particle size did not change the basic HA (Mr = 300000) the coupled particle size growth from 122.2nm to 131.3nm, indicating that the high molecular weight HA greater impact on particle size change. As can be seen by SEM photographs zoom multiples ligand coupled nanoparticle distribution, good dispersion; (3) nanoparticles release rate and the cumulative amount of drug release order for pH = 4.3 pH = 5.6> pH = 7.4, and the stronger the acidity within a certain pH range higher release rate and cumulative release, high drug loading nanoparticles release rate is higher than in the same environment in low drug loading nanoparticles FA coupling basically no effect on the drug release properties of nanoparticles, macromolecular HA coupled to the surface of the nanoparticles, drug release rate was significantly lower. Exhibit good release effect targeting PLA drug-loaded nanoparticles simulated environment in vivo release fitting FA / HA-PLA model the release of the drug-loaded nanoparticles were ININ (1 / (1-Q)) = 0.04633 Int-1.03562 and 1-Q1 / 3 =-0.01368t +1.04726-. |
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