几种中药活性成分载药纳米微球的制备及其生物活性研究

发布时间：2018-01-23 00:18

本文关键词： 厚朴酚和厚朴酚离子交联纳米微球生物活性　出处：《武汉工程大学》2015年硕士论文　论文类型：学位论文

【摘要】：厚朴酚(Magnolol)与和厚朴酚(Honokiol)是中药厚朴中的两个主要活性成分,具有良好的抗炎、抗菌、抗肿瘤、降胆固醇和抗衰老等广泛的药理活性,且毒副作用小。厚朴酚与和厚朴酚溶解性差,在潮湿、阳光、高温等条件下极易发生氧化、聚合、缩合等反应,限制了其应用。采用纳米技术,将厚朴酚、和厚朴酚与载体制备成载药纳米微球则可克服上述不足,目前该研究鲜有报道。本论文以壳聚糖为载体制备厚朴酚、和厚朴酚-壳聚糖载药纳米微球,确定最佳制备条件,对载药纳米微球的体外药物释放行为、抗氧化活性和对癌细胞的细胞毒性进行了研究。为增强所制备的载药纳米微球对癌细胞的靶向作用,将叶酸偶联到载体壳聚糖分子上制备厚朴酚-叶酸偶联壳聚糖载药纳米微球。此外,为了验证本制备方法的普适性,将药物模型换成与厚朴酚、和厚朴酚分子结构类似的白藜芦醇,制备载药纳米微球。本论文主要研究工作如下:1.厚朴酚或和厚朴酚-壳聚糖载药纳米微球制备及生物活性:采用离子交联法通过调节体系pH值、交联剂三聚磷酸钠浓度、稳定剂泊洛沙姆188浓度和乙醇的含量,确定最佳条件制备厚朴酚、和厚朴酚-壳聚糖载药纳米微球。原子力显微镜(AFM)观察纳米微球形貌和尺寸;通过紫外光谱(UV)和红外光谱(FT-IR)对纳米微球进行表征;用X射线粉末衍射(XRD)和动态光散射法(DLS)确定药物在纳米微球中的状态和微球在溶液中的分散性、稳定性;差式扫描量热法(DSC)和热重分析(TG)确定纳米微球的组成和药物的热稳定性;高效液相色谱(HPLC)测定纳米微球的载药量、包封率;紫外法测定体外药物释放曲线;二苯代苦味酰基自由基法(DPPH?)法测定纳米微球的清除自由基能力;MTT法测纳米微球对不同细胞的细胞毒性。结果表明,厚朴酚、和厚朴酚-壳聚糖载药纳米微球成球稳定、粒径均一,大小在100 nm左右,溶液中分散性较好。所制备的厚朴酚-壳聚糖载药纳米微球载药量为81.71?g/mg,包封率为20.27%;和厚朴酚-壳聚糖载药纳米微球的载药量为240.64?g/mg,包封率为59.68%。载药纳米微球有一定的缓释和控释能力,能保持药物的抗氧化活性,对多种癌细胞都有较好的抗癌活性。将异硫氰酸荧光素标记到壳聚糖分子上,以厚朴酚为药物模型制备荧光标记的厚朴酚-壳聚糖载药纳米微球研究其细胞摄取情况。结果表明,L02细胞和SMMC7721细胞都能很好的摄取载药纳米微球,且癌细胞能摄取更多的纳米微球。2.厚朴酚-叶酸偶联壳聚糖载药纳米微球的制备及其细胞毒性:将叶酸分子通过酰化反应偶联到壳聚糖分子上,以叶酸偶联壳聚糖为载体,以厚朴酚为药物模型,在最佳反应条件下制备纳米微球。对制备的厚朴酚-叶酸偶联壳聚糖载药纳米微球的形貌、尺寸、在溶液中的分散性、稳定性进行表征,测其载药量、包封率,并对厚朴酚的体外释放行为进行测试。结果表明,所制备的纳米微球成球稳定,粒径均一,大小在180 nm左右,溶液中分散较好,载药量为155.09?g/mg,包封率为38.46%。在酸性环境中释放速率低于中性环境,有一定的pH选择性,对SMMC7721细胞的细胞毒性大于L02细胞,有一定的靶向性。3.白藜芦醇-壳聚糖载药纳米微球的制备:以白藜芦醇为药物模型,以壳聚糖为载体,在最佳反应条件下制备纳米微球。对制备的白藜芦醇-壳聚糖载药纳米微球的形貌、尺寸、在溶液中的分散性、稳定性进行表征,并测其载药量、包封率。结果表明,所制备的纳米微球成球性好,粒径均一,大小在150 nm左右,溶液中分散性好,载药量达到48.81?g/mg时,包封率为11.71%。验证了本论文制备方法的普适性。
[Abstract]:Magnolol and honokiol (Magnolol) two (Honokiol) is the main active ingredient of traditional Chinese medicine Magnolia officinalis, has good anti-inflammatory, antibacterial, antitumor, anti-aging and pharmacological activity of cholesterol lowering widely, and low toxicity. Magnolol and poor solubility in the wet, sunshine. Under the condition of high temperature and prone to oxidation, polymerization, condensation reaction, limiting its application. Using nanotechnology, the magnolol, honokiol and carrier preparation of drug loaded nanoparticles can overcome the above shortcomings, the present study is rarely reported. This paper using chitosan as the carrier preparation of honokiol honokiol - drug loaded chitosan nanoparticles, determine the optimum preparation conditions, in vitro drug release behavior of drug loaded nanoparticles, antioxidant activity and cytotoxicity to cancer cells was studied. In order to enhance the drug loaded nano microspheres for targeting to cancer cells Use folic acid to be coupled to carrier chitosan prepared magnolol - folate conjugated chitosan microspheres. In addition, in order to validate the preparation method of the universal model for drug and honokiol, magnolol and similar molecular structure of resveratrol, preparation of drug loaded nano meter the microspheres. The main research work of this thesis are as follows: 1. drug loaded honokiol and magnolol - chitosan nanoparticles preparation and biological activity: by adjusting the pH value of the system by ionic crosslinking method, crosslinking agent, sodium tripolyphosphate concentration, stabilizer concentration and content of Bo Losham 188 ethanol, to determine the optimum conditions for the preparation of magnolol. Honokiol loaded chitosan microspheres. Atomic force microscopy (AFM) to observe the morphology and size of nano microspheres; by ultraviolet spectroscopy (UV) and infrared spectroscopy (FT-IR) were used to characterize the nanoparticles; X ray powder diffraction (XRD) and dynamic light scattering (DLS). Drugs in the state of nanoparticles and microspheres in solution dispersion stability; differential scanning calorimetry (DSC) and thermogravimetric analysis (TG) to determine the thermal stability and composition of nano drug microspheres; high performance liquid chromatography (HPLC) determination of drug loading nanoparticles, entrapment efficiency; Determination of in vitro drug release kinetics of UV method; two phenyl bitter acyl radical (DPPH?) method for the determination of nanoparticles of free radical scavenging capacity; MTT method was used to measure the cytotoxicity of nanoparticles to different cells. The results showed that magnolol, honokiol loaded chitosan nano microspheres, particle size, size of around 100 nm, was a good dispersion. The magnolol - preparation of chitosan microspheres loading dosage was 81.71? G/mg, the encapsulation rate was 20.27%; the drug loading of honokiol loaded chitosan nanoparticles was 240.64? G/mg, the encapsulation rate of 59.68%. drug loaded nanoparticles The ball has a sustained and controlled release ability, can maintain the antioxidant activity of drugs have good anticancer activity against various cancer cells. The fluorescein isothiocyanate labeled chitosan molecules, to honokiol honokiol shell model drug preparation of fluorescence labeled chitosan nanoparticles on the cell uptake the situation. The results showed that L02 cells and SMMC7721 cells are good uptake of drug loaded nanoparticles, and cancer cells, and the cytotoxicity of the preparation of drug loaded nanoparticles uptake more.2. magnolol - folate conjugated chitosan nanoparticles: folic acid molecular reaction coupled to chitosan by acid. The folate conjugated chitosan as carrier to magnolol as drug model, the nanospheres were prepared under the optimum reaction conditions of preparation of magnolol - folate coupled chitosan microspheres morphology, size, dispersion in solution Stability, characterization, the drug loading, encapsulation efficiency, and in vitro release behavior of magnolol were tested. The results showed that the nanoparticles into stable, uniform particle size, a diameter of about 180 nm, well dispersed in solution, the drug loading was 155.09 g/mg, the encapsulation rate? 38.46%. in the acidic environment of the release rate is lower than the neutral environment, have certain pH selective cytotoxicity to SMMC7721 cells than L02 cells have targeted.3. resveratrol chitosan microspheres prepared with resveratrol as a model drug, using chitosan as carrier, preparation the preparation of nano microspheres under the optimum reaction conditions. The morphology of resveratrol chitosan prepared chitosan microspheres size dispersed in the solution, the stability was characterized, and its drug loading, encapsulation efficiency. The results showed that the nanoparticles into good shape, uniform particle size When the size is about 150 nm, the dispersivity of the solution is good and the drug loading reaches 48.81? G/mg, the encapsulation efficiency is 11.71%., which verifies the universality of the preparation method of this paper.

【学位授予单位】：武汉工程大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TB383.1;TQ461

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