一种靶向Wnt/β-catenin通路的光动力超分子纳米药物的制备

Preparation of a photodynamic supramolecular nanodrug targeting Wnt/β-catenin pathway

目的:以维替泊芬(VER)和XAV-939为模型药物,制备一种靶向Wnt/β-catenin通路的光动力超分子纳米药物(VER/XAV-939 SMN)。方法:以粒径、多分散指数(PDI)、包封率(EE)为评价指标,优化VER/XAV-939质量比、二硬脂酰基磷脂酰乙醇胺-聚乙二醇2000(DSPE-MPEG2000)/胆固醇(Cholesterol)质量比、稳定剂(DSPE-MPEG2000+Cholesterol)/药物(VER+XAV-93)质量比、有机相(DMF)体积、搅拌温度、搅拌速度及搅拌时间。对优化处方和工艺制得的纳米粒进行质量评价,包括粒径、PDI、Zeta电位、EE、载药量(DL)、透射电镜、光谱分析、纳米粒形成机制、纳米粒稳定性等。结果:初步确定了VER/XAV-939 SMN较优的处方和工艺,VER/XAV-939质量比1∶1,DSPE-MPEG2000/Cholesterol质量比2∶1,稳定剂/药物质量比15∶1,DMF体积300μL,搅拌温度20℃,搅拌速度600 r·min-1,搅拌时间60 min。优化处方及工艺制得的纳米粒粒径为(86.04±1.35)nm, PDI为0.11±0.02,Zeta电位为(-44.39±1.69)mV,EE为(97.61±1.25)%,DL为(6.13±0.13)%;纳米粒呈球状,不粘连;VER/XAV-939 SMN具有VER和XAV-939的特征紫外吸收峰;VER/XAV-939 SMN的荧光强度弱于游离VER;VER/XAV-939 SMN形成的主要作用力是疏水相互作用;VER/XAV-939 SMN在PBS中具有较好的稳定性。结论:该文基于自组装策略构建了VER、XAV-939纳米递送体系,该体系有望实现双通道免疫刺激,增强免疫检查点抑制剂疗效。

OBJECTIVE To prepare a photodynamic supramolecular nanodrug(VER/XAV-939 SMN)targeting Wnt/β-catenin pathway using vertiporfin(VER)and XAV-939 as model drugs.METHODS The mass ratio of VER/XAV-939,mass ratio of DSPE-MPEG2000/cholesterol,mass ratio of stabilizer/drug,organic phase(DMF)volume,stirring temperature,stirring speed and stirring time were optimized using particle size,polydispersity index(PDI)and encapsulation efficiency(EE)as evaluation indexes.The quality evaluation of VER/XAV-939 SMN made by optimized prescription and process included particle size,PDI,zeta potential,EE,drug loading capacity(DL),transmission electron microscopy,spectral analysis,nanoparticle formation mechanism,and nanoparticle stability.RESULTS Preliminarily,agood prescription and process for VER/XAV-939 SMN was determined with mass ratio of 1∶1 for VER/XAV-939,mass ratio of 2∶1 for DSPE-MPEG2000/Cholesterol,mass ratio of 15∶1 for stabilizer/drug,volume of 300μL for DMF,stirring temperature of 20℃,stirring speed of 600 r·min-1,and stirring time of 60 min.The optimized prescription and process resulted in nanoparticle size of(86.04±1.35)nm,PDI of 0.11±0.02,Zeta potential of(-44.39±1.69)mV,EE of(97.61±1.25)%,and DL of(6.13±0.13)%.Nanoparticles were spherical and non-adhesive.VER/XAV-939 SMN had the characteristic UV absorption peaks of VER and XAV-939.The fluorescence intensity of VER/XAV-939 SMN was weaker than that of free VER.The main force for nanoparticle formation was hydrophobic interactions.VER/XAV-939 SMN had good stability in PBS.CONCLUSION In this study,we constructed VER and XAV-939 nano-delivery system based on an autonomous loading strategy,which was expected to achieve dual-channel immune stimulation and enhance the efficacy of immune checkpoint inhibitors.