Preparation Of Angiopep-2 Functionalized Ginsenoside-Rg3 Nanopaticles And The Study Of Anti-glioma In Virto

Glioma is one of the most common and aggressive intracranial malignancies with high morbidity and mortality.The glioma is different from other cancers due to its diffuse invasion of the surrounding normal brain tissue,makes it is impossible to complete removal of glioma by conventional surgery.Chemotherapy was essential in the auxiliary treatment,but the result was disappointed due to the existence of blood-brain barrier.Ginseng is a traditional Chinese drug,with a long medicinal history.The main active components of Ginseng are the ginsenosides,among which the ginsenoside-Rg3(Rg3)has recently emerged as a medicine with evident antitumor effects.Unfortunately,the activity of ginsenoside-Rg3 against glioma has been disappointing in clinical study because poor solubility as well as poor blood-brain barrier permeability.Combining ginsenoside-Rg3 with the rapidly developing nanoparticles and modified with specific ligands to build targeted drug delivery system,which can increase the delivery efficiency and therapeutic effect.The main content of this paper includes three parts as follows:(1)The preparation and characterization of ginsenoside-Rg3 nanoparticles(Rg3-NP);(2)The preparation and characterization of Angiopep-2 functionalized ginsenoside-Rg3 nanoparticles(ANG-Rg3-NP);(3)The effect of ANG-Rg3-NP on C6 glioma cells and the evaluation of the targeting effect.At first,nanoparticles were prepared by the emulsion solvent evaporation method and the preparation conditions of ginsenoside-Rg3 nanoparticles(Rg3-NP)were optimized by orthogonal experimental design with the indexes of encapsulation efficiency and loading capacity.The optimal preparation conditions for Rg3-NP were the concentrations of ginsenoside-Rg3,PCL-PEG and sodium cholate were 3 mg/mL,5 mg/mL and 0.5%(w/v),respectively.Also,the volume of sodium cholate was 5 mL.Secondly,Angiopep-2 was functionalized to Rg3-NP via a maleimide-thiol covalent binding reaction to obtain ANG-Rg3-NP.The ANG-Rg3-NP was observed to be uniformly spherical in shape with a particle size at 148.7±3.3 nm.The encapsulation efficiency and loading capacity reached to 75.63±3.02% and 27.23±1.36%,respectively.Also,ANG-Rg3-NP exhibited a desirable sustained release behavior and the ginsenoside-Rg3 existed in ANG-Rg3-NP with an amorphous state.Finally,in vitro cell experiments,using brain microvascular endothelial cells(BMEC)as the model of blood-brain barrier and C6 cells as the model of tumor cell,the anti-tumor efficacy of ANG-Rg3-NP was evaluated in vitro cytotoxicity study and targeting effect.What’s more,the penetration ability of ANG-Rg3-NP by using coumarin-6 as the fluorescence probe and in virto cellular uptake was performed.In vitro cytotoxicity study indicated that ANG-Rg3-NP could inhibit the proliferation of C6 glioma cells in a concentration-dependent manner.What’s more,the functionalization of Angiopep-2 made ANG-Rg3-NP acrossed the blood brain barrier more easier and accelerated the cellular uptake of nanoparticles.