【作者】 逄秀娟；

【导师】 崔福德；

【作者基本信息】 沈阳药科大学 ， 药剂学， 2010， 博士

【摘要】 磁性固体脂质纳米粒(magnetic solid lipid nanoparticles, MSLN)是将药物和磁性物质载入固体脂质纳米粒中而制成的载体制剂,在一定强度的外磁场作用下,使药物定位于靶区,浓集并释放,从而在病变部位发挥疗效,具有高效、速效、低毒的特点。该系统可以实现物理靶向给药,是肿瘤等疾病治疗的理想剂型。本文作者首先采用共沉淀法制备了纳米级Fe3O4磁流体,以其作为纳米粒中的磁性物质,选择布洛芬为难溶性药物的模型药,选择顺铂为水溶性药物的模型药,通过处方及工艺筛选制备含药磁性固体脂质纳米粒。本论文主要包括以下几个方面：第一篇油酸包裹纳米级Fe3O4制备工艺及性质考察本篇采用共沉淀法制备了Fe3O4磁流体,以生成物颜色,磁响应时间和分散性为考察指标,在单因素考察基础上,进行正交试验,确定Fe3O4最佳处方和工艺方法。并对Fe3O4粒子表面用油酸进行修饰,增加其稳定性和进入固体脂质纳米粒(SLN)脂质核心的几率。采用扫描电镜(FESEM)和透射电镜(TEM)观察纳米级Fe3O4呈球形,粒子分布均匀。粒子大小约7nm；X射线衍射图谱显示油酸包覆后的特征峰与标准Fe3O4的特征衍射峰相吻合；DSC-TGA结果说明大约7.05%的油酸包覆在Fe3O4粒子表面,且在粒子的表面形成化学吸附,这一结果同样得到FTIR的证实；在-4000～4000 Oe磁场范围下,温度为5K时,Fe3O4磁流体显示铁磁性,矫顽力为260 Oe,在温度为300K时,Fe3O4磁流体无磁滞现象,显示超顺磁性,饱和磁化强度分别为71 emu／g和63 emu／g。第二篇布洛芬磁性固体脂质纳米粒(IB-MSLN)的研究本篇采用乳化分散-超声法制备了IB-MSLN。以稳定性和包封率为指标,在单因素考察的基础上,进行正交试验,优选两个分别以硬脂酸和单硬脂酸甘油酯为脂质材料的IB-SLN处方。按照优选的IB-SLN处方,加入纳米级Fe3O4,制备了两种IB-MSLN。采用TEM观察两种IB-MSLN均呈均匀球形；采用激光粒度仪测定IB-MSLN I和IB-MSLN II的平均粒径分别为(127±17)nm和(122±16)nm；电位测定仪测定Zeta电位分别为(-1.43±0.6)mV和(-16.1±5.53)mV；用磁性超导量子磁强计(SQUID)测量IB-MSLN I居里温度为98K,磁性纳米粒在居里温度上为超顺磁,在该温度下为铁磁性,矫顽力为200 Oe。在5k和300k时的饱和磁化强度分别为10.81和9.45 emu/g. IB-MSLNⅡ居里温度为78K,矫顽力为240 Oe,在5k和300k时的饱和磁化强度分别为3.57和2.86 emu／g；采用葡聚糖凝胶微柱离心法测定了IB-MSLN包封率分别为86.9%和84.1%；用透析袋法研究了IB-MSLN体外释放特征,IB-MSLNⅠ以Higuchi、一级方程释放模型拟和最好,IB-MSLNⅡ以Higuchi释放模型拟和最好；初步考察并确定了IB-MSLNⅡ的冻干处方和工艺。以家兔卵蛋白抗原所致的关节炎作为类风湿性关节炎模型,以家兔关节肿胀度及体内抗卵清蛋白抗体效价变化为指标来评价IB-MSLNⅡ的药效。经关节腔注射后,IB-MSLNⅡ能显著抑制家兔抗原性关节炎引起的关节肿胀和家兔体内抗卵清蛋白抗体的产生。IB-MSLNⅡ的药效优于IB-SLNⅡ,基本达到了预期的目的。第三篇顺铂磁性固体脂质纳米粒(CDDP-MSLN)的研究本篇采用乳化分散-超声法制备了CDDP-MSLN。以稳定性和包封率为指标,单硬脂酸甘油酯作为脂质材料,在单因素考察的基础上,进行正交试验,优选CDDP-SLN处方。按优选处方,加入Fe304磁流体,制备了CDDP-MSLN。采用TEM观察CDDP-MSLN呈均匀球形；采用激光粒度仪和电位测定仪测定了粒径大小和Zeta电位,分别为(125±17)nm、(-13.3±6.94)mV；X射线粉末衍射和DSC结果说明顺铂在MSLN中以无定型存在,基本包入载体中；CDDP-MSLN居里温度为150K,矫顽力为310 Oe。在5k和300k时的饱和磁化强度分别为1.45和1.28 emu／g。采用葡聚糖凝胶微柱离心法测定了CDDP的包封率为75.9%；用透析袋法研究了CDDP-MSLN的体外释放特征,符合Weibull释放模型；CDDP-MSLN稳定性考察不稳定,应制成冻干品存放。初步考察并确立了CDDP-MSLN冻干处方和工艺。建立了HPLC柱前衍生化法测定顺铂体内含量。Wistar大鼠分组后静脉注射给药,考察了顺铂溶液、不加磁场和加磁场CDDP-MSLN的体内分布行为。结果表明,CDDP-MSLN在外加磁场的作用下,能够聚集于指定的靶区,靶向性显著,达到了物理靶向作用。更多还原

【Abstract】 Magnetite-loaded solid lipid nanoparticles (MSLN) represent a class of functional materials that may have potential for use in drug targeting. It is usually composed of a magnetic part and a solid lipid nanoparticles (SLN) part. The magnetic part is often an inorganic magnetite nanoparticles. The SLN part are particles made from solid lipids with a mean diameter between approximately 50 and 1000 nm. The magnetite-loaded solid lipid nanoparticles can be potentially used in drug targeting. Magnetite can be synthesized through co-precipitation of ferrous and ferric aqueous solution by addition of a base. Ibuprofen as insolubility model drug and CDDP as water-solubility model drug were studied for preparing Magnetite-loaded solid lipid nanoparticles (MSLN).The main research aspects are as follows:First Chapter:The preparation of oleic-acid-coated magnetite nanoparticles.Magnetite can be synthesized through co-precipitation of ferrous and ferric aqueous solution by addition of a base. In order to prevent them from possible oxidation in air as well as form agglomeration, the magnetite nanoparticles should be coated with oleic acid during precipitation process. At the same time that it can be increased its lipophilicity and consequently their incorporation into lipid phase. Effect of Fe3O4 magnetic nanoparticles products with different reaction temperature and concentration were investigated by single factor experiments. The optimal processes and formulation was obtained by orthogonal experiment design, based on the color, dispersibility and magnetic response time.The appearance was examined by FESEM and TEM, The results showed that oleic-acid-coated magnetite have well-defined spherical or elliptical shapes, which have a diameter of 7 nm. The XRD pattern of oleic-acid-coated magnetite is in good agreement with that of standard magnetite. The results of DSC-TGA indicate the Fe3O4 envelopment amount of oleic oil is about 7.05%, the oleic oil was chemical adsorbed on the surface of particle to form the primary layer. SQUID measurements indicated that the saturation magnetization at 5K and 300K is 71 emu/g and 63 emu/g. Second Chapter:Studies on Magnetic Solid Lipid Nanoparticles Loaded Ibuprofen (IB)The effects on the appearance and stability of IB-SLN of processes and formulations were investigated by single factor experiments. Two kind of optimal formulation were obtained by orthogonal experiment design, based on the encapsulate efficiency(EE%). Fe3O4 magnetic fluid were added optimal formulation of IB-SLN. Two kind of IB-MSLNs was finally acquired with the method of emulsification dispersion-ultrasound.The result of TEM showed that Two kind of IB-MSLNs were sphericity; the average size IB-MSLNⅠand IB-MSLNⅡwere (127±17) nm and (122±16)nm, respectively; and zeta potential were (-1.43±0.6) mV and (-16.1±5.53) mV, respectively; SQUID measurements indicated that IB-MSLN I and IB-MSLN II exhibited superparamagnetic behavior with a blocking temperature of 98 K and 78 K. The saturation magnetization at 5K and 300K for IB-MSLN I is 10.81 and 9.45emu/g and for IB-MSLNⅡis 3.57 and 2.86emu/g, respectively. the EE% of ibuprofen and ferroso-ferric oxide were 86.9% and 84.1%; the release of IB-MSLN was completely in the given mediums in 36h, which was according to Higuchi model.The formulations and processes of lyophilization were investigated basing on the appearance, color and redispersibility of IB-MSLNⅡ.IB-MSLNⅡsuspensions were administered into the cavity in a model of antigen-induced arthritic rabbit and evaluated with joint swelling and antibody titer to ovalbumin. IB-MSLN provided a conspicuous pharmacological efficacy in the joints of arthritic rabbits such as reducing joint swelling and antibody titer to ovalbumin. Ibuprofen-MSLN was better than ibuprofen-SLN in pharmacodynamics.Third Chapter:Studies on Magnetic Solid Lipid Nanoparticles Loaded CDDPThe effects on the appearance and stability of CDDP-SLN of processes and formulations were investigated by single factor experiments. The optimal formulation was obtained by orthogonal experiment design, based on the encapsulate efficiency(EE%). The Fe3O4 magnetic fluid, which was synthesized by co-precipitation method, was added in the formulation. The Magnetic Solid Lipid nanoparticles loaded CDDP (CDDP-MSLN) was finally acquired with the method of emulsification dispersion-ultrasound, with glycerin monostearate as lipid carrier, soybean phospholipids for injection, Pluronic F-68 and Tween80 as emulsifiers. CDDP-MSLN possessed a small size and an uniform particle size distribution, which were consistent with the demand of nano-preparations.The results of TEM showed that CDDP-MSLN was sphericity; the average size, zeta potential and EE% were (132±18) nm, (-13.3±6.94) mV and 75.9%, respectively; SQUID measurements indicated that CDDP-MSLN exhibited superparamagnetic behavior with a blocking temperature of 150 K. The saturation magnetization at 5K and 300K for CDDP-MSLN is 1.45 and 1.28 emu/g. The results of XRD was indicated that CDDP was embedded into MSLN with the form of amorphous; Results of release experiments showed that the release of CDDP-MSLN were completely in the given mediums, which was according to Weibull model. Besides, CDDP-MSLN was instable in 4℃and 25℃, therefore, the lyophilized products were necessary for storage.The formulations and processes of lyophilization were investigated base on the appearance, redissolution and redispersibility of CDDP-MSLN.A pre-column derivatization HPLC method was established for the in vivo assay of CDDP. Nickel chloride and diethyldithiocarbamate(DDTC) were employed as internal standard and derivatization agent, respectively. This method was proved to be specified and precise, which was fit for the in vivo assay of CDDP. Three different formulations, which included CDDP saline solution, CDDP-MSLN and CDDP-MSLN under applied magnetic field, respectively, were administrated i.v. in rats to examine their in vivo distributions. We can reach to a conclusion that CDDP-MSLN was able to gather to targeted areas and exhibited an obvious targeted effect under applied field, therefore the physical targeted effects were finally achieved.更多还原