波棱内酯A纳米混悬剂冻干粉的制备及其抗乙肝病毒活性评价

Preparation of herpetolide A nanosuspension lyophilized powder and evaluation of its anti-hepatitis B virus activity

制备波棱内酯A纳米混悬剂冻干粉(HPA-NS-LP),考察其体外溶出和抗乙肝病毒活性。采用超声沉淀法制备波棱内酯A纳米混悬剂(HPA-NS),单因素试验优化其处方及工艺;冷冻干燥法制备HPA-NS-LP。扫描电镜(SEM)观察形态;桨法测定其溶出度。以HepG2.2.15细胞模型评价HPA-NS-LP体外抗乙肝病毒活性,并与波棱内酯A普通混悬剂冻干粉(HPA-CS-LP)进行比较。结果显示,优化后的HPA-NS的粒径为(173.46±4.36)nm,PDI为0.110±0.012;制备所得的HPA-NS-LP复溶后平均粒径和PDI均有所增加,但变化值均在合理范围之内;SEM下观察HPA-NS-LP呈圆球状;体外溶出度试验发现HPA-NS-LP在2 h内累积溶出度达到90%以上,显著高于HPA-CS-LP;HPA-CS-LP与HPA-NS-LP均可有效抑制HepG2.2.15细胞抗原(HBsAg和HBeAg)的分泌,但HPA-NS-LP的抑制作用显著强于HPA-CS-LP(P<0.05)。高、中、低剂量(50,25,12.5 mg·kg-1)的HPA-NS-LP可显著抑制HepG2.2.15细胞HBV-DNA的复制(P<0.05);且效果优于同剂量的HPA-CS-LP(P<0.05)。结果表明波棱内酯A(HPA)制备成纳米混悬剂冻干粉后抗乙肝病毒活性明显增强。

To prepare the herpetolide A nanosuspension lyophilized powder(HPA-NS-LP),in order to investigate its anti-hepatitis B virus(HBV)activity and the dissolution in vitro.Herpetolide A nanosuspension(HPA-NS)was prepared by ultrasonic precipitation method.The formulation and process of HPA-NS were optimized by the single factor experiment.Lyophilized powder(HPA-NS-LP)was prepared by freeze-drying method.Scanning electron microscopy was used to observe morphology of HPA-NS-LP.Paddle method was used to determinate the dissolution of HPT-NS-LP in vitro.The anti-HBV activity of herpetolide A coarse suspension lyophilized powder(HPA-CS-LP)and HPA-NS-LP was evaluated by HepG2.2.15 cell model.The mean particle size of optimized HPA-NS was(173.46±4.36)nm,with a polydispersity index of 0.110±0.012.After redispersion,the mean particle size and the polydispersity index of HPA-NS-LP increased,with changes within a rational range.Scanning electron microscopy showed that HPA-NS-LP was spherical in shape.Cumulative dissolution rate of HPA-NS-LP was more than 90%in 2 hours,which was higher than that of HPA-CS-LP.Both HPA-CS-LP and HPA-NS-LP could effectively inhibit the secretion of HepG2.2.15 cell antigens(HBsAg and HBeAg),and the inhibitory effect of HPA-NS-LP was significantly higher than that of HPA CS-LP(P<0.05).HBV-DNA test showed that high,medium and low-dose HPA-NS-LP(50,25,12.5 mg·kg-1)significantly decreased the level of HBV-DNA(P<0.05),and the effect was better than that of the same dose of HPA-CS-LP(P<0.05).The results revealed that HPA-NS-LP exhibited anti-HBV activity in vitro,and its effect was superior to that of HPA-CS-LP.