齐墩果酸线粒体靶向脂质体的制备及抗胰腺癌研究

Preparation of oleanolic acid mitochodrial targeting liposome and its anti-pancreatic cancer effect

目的制备齐墩果酸(OA)线粒体靶向脂质体(α-TOS-TPP-OA),以实现药物的靶向性并提高其抗胰腺癌的活性。方法通过两步反应,合成三苯基膦(TPP)修饰的生育酚琥珀酸酯(α-TOS)得到线粒体靶向载体(α-TOS-TPP);采用过膜挤压法制备α-TOS-TPP-OA,并通过超滤法分离α-TOS-TPP-OA和游离药物,测得其包封率和载药量;通过动态光散射、透射电子显微镜对α-TOS-TPP-OA的粒径大小、表面电荷、形态进行表征,并通过线粒体共定位考察其线粒体靶向性;通过MTT法和流式细胞仪考察OA、α-TOS-TPP、α-TOS-TPP+OA和α-TOS-TPP-OA对胰腺癌细胞(Bxpc-3)增殖的抑制和诱导凋亡的作用。结果制备的α-TOS-TPP-OA平均粒径为(137.01±0.82)nm,PDI为(0.23±0.01),表面电荷为+(43.09±1.22)mV;α-TOS-TPP的包封率和载药量分别为(76.21±7.74)%和(39.06±5.51)%,OA的包封率和载药量分别为(70.96±9.13)%和(3.90±0.75)%;能较好地定位于线粒体;α-TOS-TPP-OA组与其他组相比抗胰腺癌的效果有明显增强。结论α-TOS-TPP-OA具有良好的稳定性和线粒体靶向性,能增强OA的抗胰腺癌活性。

Objective To prepare oleanolic acid(OA)mitochondria-targeted liposomes(α-TOS-TPP-OA)for drug targeting and improve anti-pancreatic cancer activity.Methods A mitochondrial targeting carrier(α-TOS-TPP)was synthesized from tocopherol succinate(α-TOS)modified by triphenylphosphine(TPP)by two-step reaction.Theα-TOS-TPP-OA was prepared by extrusion through the membrane.Free drugs andα-TOS-TPP-OA were separated by ultrafiltration,and the entrapment efficiency and drug loading were measured.The particle size,surface charge and morphology ofα-TOS-TPP-OA were characterized by dynamic light scattering and transmission electron microscope.The targeting property ofα-TOS-TPP-OA was determined by co-localization of mitochondria.The effects of OA,α-TOS-TPP,α-TOS-TPP+OA andα-TOS-TPP-OA on the proliferation and apoptosis of pancreatic cancer cells(BxPC-3)were investigated by MTT and flow cytometry.Results The average particle size,PDI and surface charge ofα-TOS-TPP-OA were(137.01±0.82)nm,(0.23±0.01)and+(43.09±1.22)mV,respectively.The entrapment efficiency and drug loading ofα-TOS-TPP were(76.21±7.74)%and(39.06±5.51)%,while the entrapment efficiency and drug loading of OA were(70.96±9.13)%and(3.90±0.75)%.The anti-pancreatic cancer effect ofα-TOS-TPP-OA group was much stronger than that of other groups.Conclusionα-TOS-TPP-OA has good stability and mitochondrial targeting,which can enhance the anti-pancreatic cancer activity of OA.