【作者】 隋小宇；

【导师】 祖元刚；

【作者基本信息】 东北林业大学 ， 植物学， 2012， 博士

【摘要】 辅酶Q1o(coenzyme Q10,CoQ10)是一种可以从植物中提取也可以由人体自身生产的苯醌类化合物。作为细胞内电子传递链的一部分,在机体能量代谢方面发挥着关键作用。同时它也是人体细胞膜中广泛存在的唯一内源亲脂性抗氧化剂,对于延缓衰老、提高机体免疫力都具有重大意义。但基因突变,衰老,癌症和药物等因素都可能导致人体中的辅酶Q10含量减少。在这种情况下,外源性辅酶Q10的补充是十分必要的。在临床上,辅酶Q10可以作为心脏疾病、免疫系统疾病、神经系统疾病等的治疗药品。辅酶Q10不溶于水,口服生物利用度很差。并且分子结构中含有不饱和双键,因此很不稳定,容易被空气中的氧及光线氧化和分解,受热或遇到金属离子则加速其分解,结果往往导致产品中辅酶Q10含量降低。这些性质极大地限制了它的应用。而纳米药物传输系统正是改善辅酶Q10水溶性、提高生物利用度以及防止药物结构遭到破坏的有效途径。本论文就针对这些问题进行了辅酶Q10纳米传输系统的研究,主要内容如下:1.为了增强辅酶Q10的亲水性,采用反溶剂重结晶法制备辅酶Q10微细颗粒。考察了药物浓度、体系温度、搅拌强度和搅拌时间等因素对辅酶Q10微细颗粒平均粒径的影响。优化的辅酶Q10微粒制备的工艺条件为:药物浓度50mg/ml,溶剂反溶剂体积比1:6,体系温度4℃,搅拌速度4000r/min,搅拌时间1Omin。按此工艺,可以制备得到平均粒径为1.84μm的微细颗粒。分别对产品进行X射线衍射、红外光谱分析和差示扫描量热分析等各项检测。检测结果显示,与原药相比,辅酶Q10微细颗粒的化学结构未发生改变,但熔点及晶体衍射峰强度均减弱。2.采用溶液法联合高压均质制备辅酶Q10-γ-环糊精包合物,以载药包合物的包封率及平均粒径为响应指标考察产品质量。通过单因素试验来选择适宜的因素及水平,在此基础上,采用响应面法优化制备工艺并得到了最佳的包合工艺条件:γ-环糊精与药物质量比7.76:1,超声温度40℃、超声时间28.33min,均质压力40MPa。经实际验证,得到载药包合物产品的平均包封率为48.39%,平均粒径334nm。该产品的饱和溶解度与原料相比,提高了约4.8倍。大鼠体内药代动力学研究结果显示,当口服剂量为40mg/kg时,原料药与包合物产品的AUC值分别为19.58和28.90μg.h·ml-1,生物利用度提高得到有效提高。3.采用乳化减压蒸发结合高压均质的方法制备辅酶Q10前体纳米脂质体,以包封率及平均粒径为指标考察产品质量。通过正交试验设计优化前体纳米脂质体的配方和制备工艺,其结果为:大豆卵磷脂与胆固醇的质量比4:1,卵磷脂用量100mg、脂药质量比10:1、有机相与水相体积比1:6,高压均质压力60MPa,减压蒸发温度450℃、初步乳化时间5min。以优化配方和工艺制得的前体脂质体平均粒径为241nm,重建脂质体的包封率为67.31%。进一步通过对比试验,筛选出海藻糖做为适宜的冻干保护剂,其添加量为1:2(卵磷脂与海藻糖的质量比)。获得的前体纳米脂质体粉末较为干燥且色泽均匀,质量稳定,在贮存90天后平均粒径与包封率均无明显变化。4.对辅酶Q10水包油(O/W)型注射亚微乳进行处方设计并对其大鼠体内药代动力学进行研究。采用伪三元相图法研究不同表面活性剂、助表面活性剂及Km值下空白亚微乳体系的形成条件。在优选亚微乳处方基础上,制备载药亚微乳并对其质量进行评价。通过静脉给药对辅酶Q10亚微乳大鼠体内药代动力学进行研究。结果显示,泊洛沙姆/大豆磷脂/乙醇/聚乙二醇400形成的亚微乳黏度低,粒径小,包封率高。药代动力学实验表明,辅酶Q10亚微乳经静脉注射给药后具有明显的缓释作用,平均滞留时间为6.55h。经过优选的辅酶Q10亚微乳,辅料用量少,稳定性好,制备简单,便于生产及临床使用。更多还原

【Abstract】 Coenzyme Q10(CoQ10)is a benzoquinone compound which can be produced by plant or the human body.As a component of the electron transport chain in the cell,coenzyme Q10 plays a key role in energy metabolism.Moreover,it is only lipophilic antioxidant in human body which is widely distributed in human cell membrane,and is very important to delay aging and improve immunity.However,coenzyme Q10 in the body would probably reduce because of gene mutation,aging,cancer or using drug.In this case,supplementation of the exogenous coenzyme Q10 is very necessary.In clinical practice,coenzyme Q10 is a medicine of curing heart disease,diabetes,Parkinson’s disease or other diseases.Coenzyme Q10 is insoluble in water and its oral bioavailability is poor.Furthermore,it is unstable and likely to be oxidized in the air and be decomposed in the sunshine because molecular structure of coenzyme Q10 contains unsaturated double bonds.In addition,heat or encountering metal ions would accelerate the decomposition.These factors can result in decrease of coenzyme Q10 content in products.These properties greatly limit its application.Nano-drug delivery system can improve solubility and bioavailability of coenzyme Q10,and can prevent effective the destruction of drug structure.In this paper,we will conduct relevant reseach on coenzyme Q10 nano-transmission system to solve these issues above.The major contents are as follows:1.In order to improve hydrophilia of coenzyme Q10,coenzyme Q10 micro-particles were prepared by anti-solvent crystallization method.We investigated influences of the drug solution concentration,system temperature,stirring speed and stirring time,etc.on micro-particle size.Finally get optimum conditions of micro-particles preparation:drug concentration 50mg/ml;solvent and anti-solvent volume ratio 1:6;the system temperature 4℃;stirring speed 4000r/min;stirring time 10min.In this process condition,microparticles of average diameter 1.84μm were obatained.Properties of products were characterized using X-ray diffraction(XRD),infrared spectroscopy(FT-IR),differential scanning calorimetry analysis(DSC),etc.The results show that chemical structure of coenzyme Q10 micro-particle has not changed,but the melting point and intensity of diffraction peak decreased.2.Solution method combined with high-pressure homogenization was used to prepare coenzyme Q10-y-cyclodextrin inclusion complex.Encapsulation efficiency and mean particle size of the drug inclusion complexes as indicators to examine the quality of products.Single-factor test was performed to select the appropriate factors and levels.On this basis,we optimize the preparation process of the inclusion using response surface method and the optimal results are as follows:the quality ratio of y-cyclodextrin and drug 7.76:1,the ultrasonic temperature of 40℃,ultrasonic time 28.33min,and homogeneous pressure of 40MPa.Under such conditons,we obtained the drug inclusion complex of average encapsulation efficiency of 48.39%,and average particle size of 334nm.The saturation solubility of complex product increased about 4.8times compared to raw material.In vivo pharmacokinetic studies in rats showed that AUC values of raw materials and the inclusion compound products were 19.58μg/h/ml and 28.90μg h·ml-1 when oral doses was 40mg/kg,and bioavailability was effectively enhanced.3.Emulsion and vacuum evaporation combined with high pressure homogenization method was used to prepare coenzyme Q10 precursor nanoliposomes.Encapsulation efficiency and mean particle size of the drug inclusion complexes as indicators to examine the quality of products.Orthogonal experimental was performed for optimizing preparation process and formulations of precursor nanoliposomes,the results are as follows:the quality ratio of soybean lecithin and cholesterol 4:1,usage amount of soybean lecithin 100mg，mass ratio of soybean lecithin and drug 10:1,volume ratio of the organic phase and aqueous phase 1:6,high pressure homogenization pressure 60MPa,vacuum evaporation temperature 45℃,the initial emulsification time 5min.Under such conditons,we obtained precursor liposome that its reconstruction solution’s average particle size was 241nm,and encapsulation efficiency was 67.31%.Through comparing experiments,trehalose was chosed as appropriate lyoprotectant,and its dosage was 1:2(mass ratio of lecithin and trehalose).Prepared precursor nanoliposome has good stability.There were no significant changes of the average particle size and encapsulation efficiency after 90 days of storage.4.To investigate the O/W submicron emulsion injection formulation,and investigate its pharmacokinetics in the rat.Pseudo-ternary phase diagrams were established using the water titration method.The effects of different surfactants,cosurfactants and Km values on the phase diagram were investigated,and the prescription of submicron emulsion formulation was optimized.Stability of coenzyme Q10 submicron emulsion was evaluated,and the pharmacokinetics in the rat after intravenous injection was study.We found coenzyme Q10 submicron emulsion consisting of Poloxamer188/Lipoid S100/ethanol/PEG400/water have the lower viscosity,the smaller size and the higher encapsulation efficiency.An obvious sustained-release effect of coenzyme Q10 submicron emulsion was observed after IV injection in the pharmacokinetics experiment,and mean residence time is 6.55h.The optimized coenzyme Q10 submicron emulsion consumes smaller quantities of auxiliary materials and shows good stability.Moreover,it is easy to manufacture and convenient for clinical usage.更多还原