Preparation And Evaluation Of In Vitro Antitumor Activities Of Folate-conjugated Vinblastine Sulfate Albumin Nanoparticles

Vinblastine is a kind of alkaloid extracted from Catharanthus roseus, which has anticancer bioactivity. Vinblastine started to be applied in clinical treatment for Hodgkin's disease, breast cancer, and none-small cell lung cancer etc since 1970s. Vinblastine is used in the form of sulfate (Vinblastine sulfate, VBLS) in clinical application, which has improved its water solubility. However, there still are shortcomings of short half-life and strong side effects on nerve system and gastrointestinal system.Nanotechnology applied into life science research is named nano-biotechnology, which combine nanotechnology and biology. Due to the change of diameter, nano-size materials obtain a series of qualities, such as quantum size effect, small size effect, surface and interface effects. Nanoparticles are much smaller than animal cells, which provides a new research direction for biological research. Nano drug carrier is now one of the most important research fields of nano-biotechnology, mainly focusing on targeting or local delivery of anticancer drugs. This project tried to eliminate the side effects of VBLS using nano-biotechnology and improve drug targeting and bioavailability, making VBLS a more effective anticancer drug.This project focused on the serious side effects and non-targeting feature of VBLS and prepared VBLS targeting injection formulation with desolvation technique. BSA nanoparticle preparation procedure was optimized with Central Composite Design (CCD);the morphology of BSA nanoparticle (BSANP) was characterized with various examination means; the vitro drug release property and stability of the prepared VBLS-folate conjugated BSANP (FA-BSANP-VBLS) were studied, and the anticancer activity of the FA-BSANP-VBLS was also investigated on PC-3 cancer cell line.1.The optimal conditions of preparing BSANP of 150 nm were obtained. The optimized formulation was achieved with 3.21 mg/mL BSA,2.96 mL/min of ethanol rate,4.23 mL ethanol, and 20% crosslinking degree.According to the Atomic Force Microscope (AFM), BSANP were evenly distributed and in spherical shape. The particle size obtained by AFM was in consistent with Laser Particle Size Analyzer.2.To some extent, the residual amino acid on the surface of BSANP can be controlled by adjusting the amount of crosslinking agent-glutaraldehyde, thus controlling the amount of folate affiliated. In this study,383.996μmol was affiliated to the optimized BSANP.3.Based on the experiments conducted with three different groups of FA-BSANP and different ratio of VBLS:FA-BSANP, we observed that the larger the particle was, the higher was the entrapment efficiency and loading efficiency. When it comes to the same diameter, the entrapment efficiency and loading efficiency increased as the ratio of VBLS: FA-BSANP increased.At 156.2 nm (optimized group), the maximal entrapment efficiency and loading efficiency were 86.93% and 44.24%, respectively.4.When the FA-BSANP-VBLS (with mannitol) freeze-dried powder was redissoluted in deionized water, the solution can remain stable and clear for at least 48 h at 4℃.5.The drug carrier system had a burst release in 12 hours and a slow release up to 56 hours, releasing approximately 30% and 80% of VBLS respectively. The observed release curve indicates that the drug carrier system had a sustained release property. Moreover, the observed release curve was fitted with software, which indicated that the observed release model was in close agreement with the first order kinetics model, confirming the sustained release feature of this drug delivery system.6.Comparing with the VBLS and BSANP-VBLS groups, FA-BSANP-VBLS demonstrated the best anticancer activity in cell apoptosis experiments. Moreover, both BSANP-VBLS and FA-BSANP-VBLS showed the feature of sustained release in the results of cell apoptosis experiments.According to the experiment results of this study, the prepared FA-BSANP-VBLS was in nanoscale and evenly distributed.They also had a narrow size distribution and a significant improvement in anticancer activity. This study is considered as an experimental foundation for further research on the in vivo activity, target ability and metabolism of the drug carrier system...