Preparation, Characterization And Evaluation Of In Vivo And In Vitro Antitumor Activities Of DHA-conjugated BSA Docetaxel- Loaded Tumor-targeting Nanoparticles

Docetaxel is a semi-synthetic taxane derivative, and as one of taxane, its antitumor effect is as same as taxol. Studies showed that docetaxel has a broad spectrum of activity against a variety of tumors, such as breast cancer, ovarian cancer and non-small lung cancer. What is more, its antitumor effect is better than taxol. Despite the antitumor effect, docetaxel have some shortcomings, for example, poor water solubility, little targeting action, hypersensitivity and toxicity.Based on these shortcomings, this study developped a novel nanoparticles whose carrier was bovine serum albumin using a emulsification method. The optimum conditions were obtained through single factor test with the 7 factors of BSA concentration, docetaxel concentration, chloroform and water volume ratio, homogenate speed, homogenate time, homogenization pressure and homogenization time with the particle size used as the index. The results showed that the optimum preparation conditions were BSA concentration with 0.04mg/mL, docetaxel concentration with 0.04mg/mL, chloroform and water volume ratio with 1:13, homogenate speed with 400rpm, homogenate time with 6min, homogenization pressure with 600bar and homogenization time with 8min, and on this condition, the particle size was 156.7±5.4 nm, the zeta potential was -32.37±1.95 mV, the drug entrapment was 10.7%±0.4%, the loading efficiency was 98.6%±9.3%, and the docosahexaenoic acid content was 4.58%±0.2%.It studied the character of DTX-DHA-BSA-NPs by scanning electron microscopy, X-ray diffractometer and differential scanning calorimeter, which showed the nanoparticles was near-spherical in shape and docetaxel was in a disordered or amorphous crystalline phase in the solid solution state. The study also showed that DTX-DHA-BSA-NPs was stable and released in a sustained manner.It indicated that DTX-DHA-BSA-NPs had targeting action by the cellular uptake assay with MCF-7. In addition, no matter in 48h (0.029μM) or 72h (0.012μM), the IC50 of DTX-DHA-BSA-Nps was lower than DTX-BSA-Nps and DTX, it indicated that DTX-DHA-BSA-NPs had better antitumor effect in vitro than DTX-BSA-NPs and DTX by MTT assay used murine lewis lung carcinoma cells LLC, human breast tumor cells MCF-7, human lung carcinoma cells A549, human gastric cancer cells SGC7901, human gallbladder cancer cells GBC-SD, human cholangiocarcinoma cancer cells RBE.In order to study the antitumor effect in vivo of DTX-DHA-BSA-NPs, it made two tumor mouse model, one of the model was made by transplanted ascitic tumor S180 to Kuming mice, the other model was made by transplanted murine lung carcinoma cells LLC to C57BL/6 mice, which showed that the antitumor effect in vivo of DTX-DHA-BSA-NPs was better than DTX-BSA-NPs and DTX.In short words, the DTX-DHA-BSA-NPs which was prepared by emulsification method had better stability, better controlled release in vitro, little toxicity, better targeting action, moreover, its antitumor effect was better than DTX-BSA-NPs and DTX no matter in vivo or in vitro.