单克隆抗体OX26修饰的丹酚酸B/黄芩苷纳米结构脂质载体研究

目的制备转铁蛋白受体单克隆抗体OX26修饰的丹酚酸B/黄芩苷纳米结构脂质载体(Sal B/BA-NLC),并对其进行表征和细胞摄入研究。方法采用乳化-固化法制备Sal B/BA-NLC,用2-亚氨基硫烷盐酸盐(2-iminothiolane hydrochloride)将OX26进行硫醇化后连接于Sal B/BA-NLC表面,以形态、粒径、Zeta电位及包封率等为评价指标考察其理化性质,并用差示扫描量热法(DSC)与核磁共振波谱(NMR)进行验证。以香豆素-6(coumarin-6,C6)为荧光探针,代替黄芩苷和丹酚酸B制备制剂,利用高内涵成像仪考察脑微血管内皮细胞bEnd.3对其的摄入情况。结果所制备的OX26修饰的Sal B/BA-NLC粒径为(27.50±3.37)nm,PDI为0.39±0.04,Zeta电位为(-7.06±1.85)m V。DSC与NMR结果表明,药物是以无定形的形式存在于纳米结构脂质载体中。细胞摄入结果显示,当考察时间一致时,bEnd.3细胞摄入3组溶液体系的荧光强度为OX26-C6-NLCC6-NLCC6-SL。结论所制备的OX26修饰的Sal B/BA-NLC粒径较小、分布均匀,且包封率高。与溶液组和未修饰的NLC组相比,OX26修饰的NLC组具有更高的摄入量。

Research on monoclonal antibody OX26 modified salvianolic acid B/baicalin nanostructured lipid carriers

Objective To prepare, characterize, and study cellular uptake of transferrin receptor monoclonal antibody OX26 modified nanostructured lipid carrier loaded with salvianolic acid B and baicalin (Sal B/BA-NLC). Methods Sal B/BA-NLC was prepared by emulsification-solvent evaporation method. OX26 was thiolated with 2-iminothiolane hydrochloride and then conjugated to the surface of Sal B/BA-NLC. The morphology, particle size, Zeta potential, and encapsulation efficiency (EE) were evaluated for the physicochemical properties, and OX26 modified Sal B/BA-NLC was verified by differential scanning calorimetry (DSC) and nuclear magnetic resonance spectroscopy (NMR). Coumarin-6 (C6) was used as the fluorescent probe instead of baicalin and salvianolic acid B to prepare the formulations in cellular uptake study. The cellular uptake study was conducted by brain microvascular endothelial cells bEnd.3 using high content cell imaging analysis system. Results The prepared OX26 modified Sal B/BA-NLC had particle size of (27.50 ±3.37) nm, PDI of 0.39 ±0.04, and Zeta potential of (-7.06 ±1.85) mV. The DSC and NMR results indicated that the drug was encapsulated in the nanostructured lipid carrier in an amorphous form. The results of cell uptake showed that the fluorescence intensities of the three solutions in bEnd.3 cells were:OX26-C6-NLC > C6-NLC > C6-SL. Conclusion The prepared OX26 modified Sal B/BA-NLC has smaller particle size, uniform distribution, and high EE. The OX26-modified NLC group had a higher intake than the solution group and the unmodified NLC group.