Affiliation of Author(s):[1]西南交通大学生命科学与工程学院,四川成都610031;[2]四川省医学科学院·四川省人民医院,药学部,四川成都610072;[3]电子科技大学医学院,个体化药物治疗四川省重点实验室,四川成都610072
Journal:Chinese Traditional and Herbal Drugs
Key Words:gambogic acid;rapamycin;co-loaded liposomes;anti-tumor;drug combination;in vivo pharmacokinetics;biodistribution;proliferation;apoptosis;migration;invasion;autophagy
Abstract:Objective To prepare liposomes co-loaded with gambogic acid and rapamycin(GR@Lip),optimize its prescription,perform anti-tumor mechanism research in vitro,pharmacokinetics and biodistribution in vivo.Methods The entrapment efficiency,drug loading,particle size were used as evaluation indicators.Single factor investigation method and orthogonal design experiments were used to optimize the optimum formulation of GR@Lip,and its characterization and stability were studied.The effects of GR@Lip on the proliferation and apoptosis of tumor cells were investigated by CCK-8 and flow cytometry.The effects of GR@Lip on the migration and invasion of tumour cells were investigated by cell scratching,Transwell.The effects of GR@Lip on the autophagy of tumour cells were investigated by transmission electron microscope(TEM)and immunofluorescence.The effects of GR@Lip on the pharmacokinetics and biodistribution in vivo were investigated by liquid chromotography with mass spectrometry(LC-MS)and living body imager.Results The optimal conditions were determined as follows:temperature is 40℃,ratio of drug to lipid is 1:1:20,the phospholipid-cholesterol ratio is 4:1,ultrasonic power is 195 W,ultrasonic time is 5 min,hydration medium is ultrapure water and aqueous pH value is 7.1.The entrapment efficiency and drug loading of gambogic acid and rapamycin were(97.27±2.76)%,(96.58±3.82)%,(3.29±0.44)%and(4.91±0.44)%,respectively.GR@Lip showed a spherical under TEM with an average particle size of(157.19±1.74)nm,ζpotential was(?22.1±1.3)mV by dynamic light scattering(DLS),and had good stability.In vitro antitumor activity experiment showed that gambogic acid combination rapamycin can inhibit the proliferation,migration and invasion of tumor cells,and also significantly promote apoptosis and autophagy.In addition,pharmacokinetics and biodistribution results in vivo showed that GR@Lip can remain for long time and had good tumor targeting.Conclusion This study successfully prepare GR@Lip,revealing that it has synergistic anti-tumor activity through multiple pathways,and significantly improving the pharmacokinetic behavior of gambogic acid and rapamycin in vivo,providing a basis for further in vivo research and future clinical applications.
Volume:55
Issue:9
Page Number:2896-2911
ISSN No.:0253-2670
Translation or Not:no
CN No.:12-1108/R