叶酸-壳聚糖修饰的7-乙基-10-羟基喜树碱脂质体的研究
发布时间：2018-11-03 10:56
【摘要】：7-乙基-10-羟基喜树碱(SN-38)是临床上抗肿瘤药伊立替康(CPT-11)的活性代谢产物,SN-38具有活性高、活性强的特点,但一些因素限制了其临床应用,如SN-38水溶性差,口服生物利用度低,体内消除较快。本文采用薄膜-探超法制备普通脂质体,再以叶酸、壳聚糖为原料合成叶酸-壳聚糖偶联物(FA-CS),然后通过吸附作用加载于脂质体表面,制备成FA-CS包裹的脂质体(FA-CS-SN-38-LP)。一方面,该制剂解决了药物难溶于水的难题;另一方面该制剂具有缓释和靶向的效果,可为SN-38的进一步开发和应用提供依据。通过单因素考察和正交试验,确定SN-38脂质体(SN-38-LP)、FA-CS包裹的脂质体(FA-CS-SN-38-LP)的处方和工艺,并以制剂的粒径、zeta电位、载药量、包封率、体外释放及稳定性等为评价指标,对制剂的性质进行表征。优化后,SN-38-LP和FA-CS-SN-38-LP的平均粒径分别为158nm和239nm,Zeta电位分别为-7.15mV和㧏18.63m V。采用超滤-离心法测定脂质体的包封率为79.23%,载药量为3.85%。采用透析法研究体外释放,比较SN-38-LP和FA-CS-SN-38-LP的释放特性。结果表明,与SN-38溶液剂(SN-38-Sol)和SN-38--LP相比,FA-CS-SN-38-LP的释放较慢,此外,SN-38-LP、FA-CS-SN-38-LP均属于脂质体载药体系,体外释放曲线相似。在体外细胞毒性研究中,以原料药为对照,MTT法考察不同浓度制剂(SN-38-LP、FA-CS-SN-38-LP)对肿瘤细胞(MCF-7、HepG-2)的抑制作用。在给药24h、48h、72h的情况下,FA-CS-SN-38-LP组对细胞的抑制作用均优于SN-38-Sol、SN-38-LP组。细胞抑制程度与制剂中药物的浓度成正相关。FA-CS-SN-38-LP对MCF-7细胞的抑制作用强于HepG-2细胞,该制剂对MCF-7细胞有较好抑制效果。建立S180荷瘤小鼠模型,考察制剂FA-CS-SN-38-LP在小鼠体内的组织分布情况。在体内抗肿瘤活性研究中,主要以重量抑瘤率和体积抑瘤率为评价指标。实验结果显示,FA-CS-SN-38-LP组重量抑瘤率和体积抑瘤率分别为67.04%和65.73%;与生理盐水组相比,SN-38-LP组和FA-CS-SN-38-LP组肿瘤生长较慢,且肿瘤表面无溃烂、出血现象。病理学检查结果显示,相比生理盐水组,FA-CS-SN-38-LP组肿瘤细胞排列疏松,呈不同程度的退变,肿瘤细胞数量减少,说明该制剂可较大程度地抑制肿瘤细胞的生长,具有较好的抗肿瘤效果。
[Abstract]:7-ethyl-10-hydroxycamptothecin (SN-38) is an active metabolite of the antitumor drug CPT-11. SN-38 has the characteristics of high activity and strong activity, but some factors limit its clinical application. For example, SN-38 has poor water solubility, low oral bioavailability and rapid elimination in vivo. In this paper, the common liposomes were prepared by thin-film method, then folic acid and chitosan were used as raw materials to synthesize folic acid-chitosan coupling compound (FA-CS), and then loaded on the surface of liposomes by adsorption. FA-CS encapsulated liposomes (FA-CS-SN-38-LP) were prepared. On the one hand, the preparation solves the problem that the drug is insoluble in water, on the other hand, it has the effect of slow release and targeting, which can provide the basis for the further development and application of SN-38. The formulation and process of SN-38 liposome (SN-38-LP) and FA-CS encapsulated liposome (FA-CS-SN-38-LP) were determined by single factor investigation and orthogonal test. The preparation size, zeta potential, drug loading capacity and encapsulation efficiency were determined. The properties of the preparation were characterized by in vitro release and stability. The average particle sizes of SN-38-LP and FA-CS-SN-38-LP were-7.15mV and-18.63 MV, respectively, and the average particle size of 158nm and FA-CS-SN-38-LP were-7.15mV and -18.63 MV, respectively. The entrapment efficiency of liposome was 79.23 by ultrafiltration-centrifugation, and the drug loading was 3.85. The release characteristics of SN-38-LP and FA-CS-SN-38-LP in vitro were studied by dialysis. The results showed that the release of FA-CS-SN-38-LP was slower than that of SN-38 solution (SN-38-Sol) and SN-38--LP. In addition, SN-38-LP,FA-CS-SN-38-LP belonged to liposome drug delivery system. In vitro release curve is similar. In vitro cytotoxicity study, the inhibitory effects of different concentrations of SN-38-LP,FA-CS-SN-38-LP on tumor cells (MCF-7,HepG-2) were investigated by MTT. The inhibitory effect of FA-CS-SN-38-LP group on cells was better than that of SN-38-Sol,SN-38-LP group at 24 h and 48 h / 72 h. The inhibitory effect of FA-CS-SN-38-LP on MCF-7 cells was stronger than that on HepG-2 cells, and the inhibitory effect of FA-CS-SN-38-LP on MCF-7 cells was better than that on HepG-2 cells. The model of S 180 tumor bearing mice was established to investigate the tissue distribution of FA-CS-SN-38-LP in mice. In the study of antitumor activity in vivo, the weight inhibition rate and volume inhibition rate were used as the evaluation indexes. The results showed that the weight inhibition rate and volume inhibition rate of FA-CS-SN-38-LP group were 67.04% and 65.73%, respectively. Compared with normal saline group, SN-38-LP group and FA-CS-SN-38-LP group had slower tumor growth, and no ulceration and bleeding on tumor surface. Pathological examination showed that compared with normal saline group, the tumor cells in FA-CS-SN-38-LP group were loosely arranged, with varying degrees of degeneration, and the number of tumor cells decreased, indicating that the preparation could inhibit the growth of tumor cells to a large extent. It has good antitumor effect.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R943
本文编号：2307584
[Abstract]:7-ethyl-10-hydroxycamptothecin (SN-38) is an active metabolite of the antitumor drug CPT-11. SN-38 has the characteristics of high activity and strong activity, but some factors limit its clinical application. For example, SN-38 has poor water solubility, low oral bioavailability and rapid elimination in vivo. In this paper, the common liposomes were prepared by thin-film method, then folic acid and chitosan were used as raw materials to synthesize folic acid-chitosan coupling compound (FA-CS), and then loaded on the surface of liposomes by adsorption. FA-CS encapsulated liposomes (FA-CS-SN-38-LP) were prepared. On the one hand, the preparation solves the problem that the drug is insoluble in water, on the other hand, it has the effect of slow release and targeting, which can provide the basis for the further development and application of SN-38. The formulation and process of SN-38 liposome (SN-38-LP) and FA-CS encapsulated liposome (FA-CS-SN-38-LP) were determined by single factor investigation and orthogonal test. The preparation size, zeta potential, drug loading capacity and encapsulation efficiency were determined. The properties of the preparation were characterized by in vitro release and stability. The average particle sizes of SN-38-LP and FA-CS-SN-38-LP were-7.15mV and-18.63 MV, respectively, and the average particle size of 158nm and FA-CS-SN-38-LP were-7.15mV and -18.63 MV, respectively. The entrapment efficiency of liposome was 79.23 by ultrafiltration-centrifugation, and the drug loading was 3.85. The release characteristics of SN-38-LP and FA-CS-SN-38-LP in vitro were studied by dialysis. The results showed that the release of FA-CS-SN-38-LP was slower than that of SN-38 solution (SN-38-Sol) and SN-38--LP. In addition, SN-38-LP,FA-CS-SN-38-LP belonged to liposome drug delivery system. In vitro release curve is similar. In vitro cytotoxicity study, the inhibitory effects of different concentrations of SN-38-LP,FA-CS-SN-38-LP on tumor cells (MCF-7,HepG-2) were investigated by MTT. The inhibitory effect of FA-CS-SN-38-LP group on cells was better than that of SN-38-Sol,SN-38-LP group at 24 h and 48 h / 72 h. The inhibitory effect of FA-CS-SN-38-LP on MCF-7 cells was stronger than that on HepG-2 cells, and the inhibitory effect of FA-CS-SN-38-LP on MCF-7 cells was better than that on HepG-2 cells. The model of S 180 tumor bearing mice was established to investigate the tissue distribution of FA-CS-SN-38-LP in mice. In the study of antitumor activity in vivo, the weight inhibition rate and volume inhibition rate were used as the evaluation indexes. The results showed that the weight inhibition rate and volume inhibition rate of FA-CS-SN-38-LP group were 67.04% and 65.73%, respectively. Compared with normal saline group, SN-38-LP group and FA-CS-SN-38-LP group had slower tumor growth, and no ulceration and bleeding on tumor surface. Pathological examination showed that compared with normal saline group, the tumor cells in FA-CS-SN-38-LP group were loosely arranged, with varying degrees of degeneration, and the number of tumor cells decreased, indicating that the preparation could inhibit the growth of tumor cells to a large extent. It has good antitumor effect.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R943
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相关期刊论文 前1条
1 许贤会;张要齐;孙雪峰;李华日;党丽梅;李会芳;刘丹;李引乾;;阿维菌素纳米脂质体的制备及品质控制[J];西北农业学报;2014年10期
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