Preparation Of Doxorubicin Hydrochloride Nanoliposome And Its In Vitro Antitumor Activity

ObjectiveTo optimize the preparation method and recipe technic of Doxorubicin Nanoliposome-Hydrochloride (DONP-HCL). To conduct detailed research on its in vitro antitumor activities and to make a systematic comparison with free DOX solution. Finally to lay a good prophasic experiment basis for this product's clinical application and further development. Method1. Liposome was prepared by choosing a certain rate of egg PC and cholesterol compound as drug DOX's carrier, and tween-80 was added into the recipe to increase solubility of DONP-HCL. Additionally, Vit-E was selected as stabilizer to increase drug stability. In this experiment, we adopt a new type of ethanol injection-PH gradient-motor mixing method to improve drug's encapsulation efficiency and control the liposome nanoparticle size; 2. HepG-2,MCF-7,Hela,SGC,U251 five cell lines were selected for screening antitumor activities of prepared DONP-HCL by MTT test; 3. Using AO-EB cell fluorescent double staining method to observe the influence of DONP-HCL on various tumor cells' apoptosis forms。ResultThe new pattern of DONP-HCL prepared in our experiment had the uniform distributed size, within 140～170nm, its entrapment rate reached as high as 99.85% and it was relatively stable in low temperature storage. The MTT test results indicated that the IC50 of DONP-HCL on HepG-2,MCF-7,Hela,SGC,U251 were 0.055μg/mL,0.8717μg/mL,1.2256μg/mL 1.5601μg/mL,1.6061μg/mL respectively, while the IC50 of free DOX solution with the same concentration on these cells were 0.932μg/mL,1.4187μg/mL,2.1332μg/mL,2.6234μg/mL,2.7671μg/mL correspondingly. Their in vitro antitumor activity and cytotoxicity existed significant differences. After treating by Dox-HCL solution and its Nano-liposome solution, lots of toruloid apoptoti,death cells could be observed using AO-EB cell fluorescent double staining method, while the negative control group appeared less cell apoptosis. And the number of living cells in DONP-HCL group was far more less than the free DOX-HCL solution group. ConclusionThe DONP-HCL prepared by ethanol injection-PH gradient-motor mixing method possessed the advantage of uniform distributed size,high encapsulation efficiency,big drug loading rate and simple craft process. The in vitro cell experiment and cell morphology observation results showed that DOX-HCL encapsulated in liposome could be more effective in enhancing the cell toxicity and promoting tumor cell apoptosis.