眼用盐酸洛美沙星壳聚糖纳米粒的制备与体外评价

Preparation and in vitro evaluation of ophthalmic lomefloxacin hydrochloride chitosan nanoparticles

目的基于质量源于设计(QbD)理念制备盐酸洛美沙星壳聚糖纳米粒(LF-CNs),并评价其体外抗菌活性。方法采用离子凝胶化法制备LF-CNs,根据失效模式效应分析(FMEA)工具初步评估影响LF-CNs制剂性质的潜在处方和工艺变量,并通过2水平部分析因实验设计筛选出影响LF-CNs制剂性质的关键变量,最终以壳聚糖浓度、三聚磷酸钠(STPP)浓度和搅拌速度3个独立变量作为考察因素,以LF-CNs的粒径分布和包封率作为评价指标,应用Box-Behnken实验设计优化并得到LF-CNs的处方和制备工艺参数。通过Zetasizer Nano ZS 90动态激光粒度仪测定LF-CNs的粒径分布、聚合物分散性指数(PDI)和ζ电位,透射电子显微镜下观察LF-CNs的微观形态;采用体外透析法考察LF-CNs和LF原料药体外药物释放特性;通过抑菌实验比较LF原料药与LF-CNs的体外抑菌活性。结果经实验优化得到LF-CNs的最优处方和工艺参数:壳聚糖质量浓度为10 mg/mL,STPP质量浓度为6 mg/mL,搅拌速度为750 r/min;在透射电镜下可观察到LF-CNs呈圆球状,无聚集,粒径大小为(479.6±18.7)nm,PDI为(0.194±0.012),ζ电位为(34.4±1.9)mV,LF-CNs的体外释放机制符合Korsmeyer-Peppas方程拟合,药物的释放为扩散和溶蚀双重机制;LF-CNs对金黄色葡萄球菌的体外抑菌效果优于LF原料药。结论将盐酸洛美沙星制备成壳聚糖纳米粒,处方设计科学合理,制备工艺简单易行,体外抑菌活性显著,有望成为盐酸洛美沙星眼用给药的一种有效途径。

Objective To prepare lomefloxacin hydrochloride chitosan nanoparticles(LF-CNs) based on the concept of quality by design(QbD), and evaluate its in vitro antibacterial activity. Methods LF-CNs were prepared by ionic gelation method. Using the failure mode effect analysis(FMEA) tool to identify the potential formulation and process variables that affect the properties of LF-CNs. The key variables that affect the properties of LF-CNs were screened by 2-level factorial experiment design. Finally, the three independent variables of chitosan concentration, STPP concentration and stirring speed were used as the investigating factors,and the particle size distribution and encapsulation efficiency of LF-CNs were used as evaluation indicators. The formulation and process parameters of LF-CNs were optimized and obtained using Box-Behnken experimental design. The particle size distribution,ζ potential and in vitro drug release characteristics of LF-CNs were investigated. The antibacterial activities of LF raw materials and LF-CNs in vitro were compared through the inhibition zone experiment. Results The optimal formulation and process parameters of LF-CNs were optimized by experiment: The chitosan concentration was 10 mg/mL, the STPP concentration was 6 mg/mL, and the stirring speed was 750 r/min. Under the transmission electron microscope, it could be observed that LF-CNs were spherical, without aggregation, the particle size was(479.6 ± 18.7) nm, the PDI was(0.194 ± 0.012), the ζ potential was(34.4 ± 1.9) mV, and the in vitro release rate of LF-CNs can be fitted by Korsmeyer-Peppas equation. The in vitro antibacterial effect of LF-CNs on Staphylococcus aureus was higher than that of LF raw materials. Conclusion In this study, lomefloxacin hydrochloride was prepared into chitosan nanoparticles. The formulation design was scientific and reasonable, the preparation process was simple and easy, and the antibacterial activity in vitro was significant. It is expected to become an effective way of ophthalmic administration of lomefloxacin hydrochloride.