酮康唑-姜黄素载药纳米系统抗菌活性协同作用的研究

发布时间：2018-06-21 01:29

本文选题：MPEG-PCL + KCZ-M　；参考：《济南大学》2017年硕士论文

【摘要】：白色念珠菌感染能够引起诸多感染性疾病,威胁人类的生命健康。多重耐药真菌及生物膜的生成,导致真菌对抗真菌药物的敏感性降低。如何提高抗真菌药物的抗菌效果,已成为目前研究的热点之一。酮康唑(Ketoconazole,KCZ)为唑类广谱抗真菌药物,对于念珠菌、各种深部真菌引起的感染均有良好治疗效果,但酮康唑严重的肝毒性限制了其临床应用。减少酮康唑用药剂量,提高其疗效成为亟待解决的重要任务。姜黄素(Curcumin,CUR)是一种安全的天然化合物,已有研究表明姜黄素能够与唑类抗真菌药物联合应用提高抗白色念珠菌能力。但酮康唑与姜黄素为疏水性药物,生物利用率低,限制了两者在临床方面的应用。聚合物胶束载药系统能够克服药物水溶性差、物理化学性质不稳定等问题,在降低药物的毒副作用同时,药物能较长时间维持在有效浓度范围内,对传统抗真菌药物的临床研究与应用起推动作用,具有潜在的研究意义。本文合成不同分子量的单甲氧基聚乙二醇-聚己内酯(Methoxy poly(ethylene glycol)-b-poly(e-caprolactone),MPEG-PCL)两亲性嵌段共聚物,结构经核磁共振氢谱(1H-NMR)及红外光谱(FT-IR)进行了确证,并经凝胶渗透色谱(GPC)测定不同PCL链段聚合物分子量。薄膜水化法制备KCZ载药胶束(KCZ-M),对影响载药胶束包封率和载药量的单因素进行考察。在单因素考察结果基础上,经正交设计筛选出最佳胶束制备工艺。研究结果表明,当MPEG/PCL重量比为1:1.7时,聚合物材料70 mg,原料药10 mg,水化温度60°C,水化时间30 min,得到KCZ载药纳米胶束,载药量与包封率最高,分别为85.27±1.75%、11.31±1.05%,CUR载药纳米胶束(CUR-M)的载药量与包封率分别为91.19±0.83%、11.90±0.11%。KCZ载药胶束的水溶性比KCZ原料药相比提高85倍,CUR载药胶束的水溶性比CUR原料药提高了82900倍,水溶性得到明显改善。采用动态光散射技术(DLS)及透射电镜(TEM)对酮康唑及姜黄素载药胶束进行了表征。酮康唑与姜黄素载药纳米胶束粒径小且分布均匀,平均粒径分别为44.70±1.91 nm、39.56±0.19 nm。傅里叶红外光谱(FT-IR)、X-射线衍射(XRD)对载药胶束进行表征,研究发现KCZ与CUR成功包裹进纳米胶束,并以无定型、非结晶形式存在。动态透析体外释放实验表明,载药胶束能控制药物的释放,药物从载药胶束中的释放呈现缓慢释放行为。KCZ-M体外释放动力学符合一级动力学模型,药物释放速度与药物浓度成正比。CUR-M体外释放动力学符合零级动力学模型,药物释放量与时间成正比。采用棋盘法和抑菌圈法对姜黄素及酮康唑载药胶束联合用药开展了体外抑菌活性实验。实验结果表明,低剂量的CUR-M(1.40~22.5μg/m L)即可改善KCZ-M的抗菌活性,二者的联合抑菌指数FICI仅为0.073,表现出明显的协同抗菌作用。以白色念珠菌为实验对象,XTT减低法以及激光共聚焦显微镜法对复配制剂抑膜活性的进一步研究表明,CUR-M可明显提高KCZ-M的抑膜活性。总之,MPEG-PCL作为酮康唑-姜黄素联合用药的潜在药物运输系统的研究,为其他抗真菌药物的研究提供了新的思路,具有潜在的临床应用价值和发展前景。
[Abstract]:Candida albicans infection can cause many infectious diseases and threaten human life and health. The formation of multidrug-resistant fungi and biofilms leads to the reduction of fungal susceptibility to fungal drugs. How to improve the antibacterial effect of antifungal drugs has become one of the hotspots of current research. Ketoconazole (KCZ) is a broad spectrum of azolic resistance. Fungal drugs have good therapeutic effects on Candida and the infection caused by various deep fungi, but the severe liver toxicity of ketoconazole restricts its clinical application. Reducing the dose of ketoconazole and improving its efficacy have become an important task to be solved. Curcumin (Curcumin, CUR) is a safe natural compound. Lutein can be combined with azolic antifungal agents to improve the ability to resist Candida albicans. But ketoconazole and curcumin are hydrophobic drugs with low bioavailability, which limits their clinical applications. Polymer micellar drug loading systems can overcome the problems of poor solubility in water and instability in physicochemical properties, and reduce drug toxicity. At the same time, the drug can be maintained within the range of effective concentration for a long time. It plays an important role in the clinical research and application of traditional antifungal drugs, and has potential research significance. In this paper, the two affinity of Methoxy poly (ethylene glycol) -b-poly (e-caprolactone), MPEG-PCL) was synthesized with different molecular weights. The block copolymer was confirmed by nuclear magnetic resonance hydrogen spectrum (1H-NMR) and infrared spectroscopy (FT-IR), and the molecular weight of different PCL chain segments was determined by gel permeation chromatography (GPC). The KCZ carrier micelles (KCZ-M) were prepared by the film hydration method. The single factor affecting the encapsulation efficiency and the drug loading of the drug loaded micelles was investigated. The basis for the single factor investigation was based on the results of single factor investigation. The optimum micelle preparation process was screened by orthogonal design. The results showed that when the weight ratio of MPEG/PCL was 1:1.7, the polymer material was 70 mg, the API was 10 mg, the hydration temperature was 60 [C], and the hydration time was 30 min, and the nano micelles loaded with KCZ were obtained. The drug loading and encapsulation rate were the highest, and were 85.27 + 1.75%, 11.31 + 1.05%, CUR carrying nano micelle (CUR-M). The drug loading and encapsulation efficiency were 91.19 + 0.83% respectively, and the water solubility of the 11.90 + 0.11%.KCZ drug micelles was 85 times higher than that of the KCZ. The water solubility of the CUR loaded micelles was 82900 times higher than that of the CUR, and the water solubility was obviously improved. The dynamic light scattering (DLS) and transmission electron microscopy (TEM) were used to carry out the drug micelles of ketoconazole and curcumin. The particle size of the ketoconazole and curcumin nanoparticles was small and evenly distributed, the average particle size was 44.70 + 1.91 nm, 39.56 + 0.19 nm. Fourier infrared spectroscopy (FT-IR), and X- ray diffraction (XRD) was used to characterize the drug loaded micelles. The study found that KCZ and CUR were successfully encapsulated into nanoscale micelles and existed in the form of amorphous and non crystalline form. In vitro release experiments show that drug loading micelles can control the release of drugs. The release of drugs from the drug loaded micelles shows a slow release behavior of.KCZ-M in vitro release kinetics in line with the first order kinetic model. Drug release rate and drug concentration are proportional to the.CUR-M release kinetics model, drug release and time. The antibacterial activity of curcumin and ketoconazole drug loaded micelles in vitro was carried out by chessboard method and bacteriostasis method. The experimental results showed that the low dose of CUR-M (1.40~22.5 g/m L) could improve the antibacterial activity of KCZ-M, and the combined antibacterial index of the two was only 0.073, which showed obvious synergistic antibacterial effect. White The further study on the inhibitory activity of the compound preparation by the XTT reduction method and the laser confocal microscope shows that CUR-M can obviously improve the inhibitory activity of KCZ-M. In a word, the research of MPEG-PCL as a potential drug transport system for the combination of ketoconazole and curcumin provides a new study for other antifungal drugs. The train of thought has potential clinical application value and development prospect.
【学位授予单位】：济南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R943

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