Dadparvar Miriam, Wagner Sylvia, Wien Sascha, Worek Franz, von Briesen Hagen, Kreuter Jörg
Institute of Pharmaceutical Technology, Johann Wolfgang Goethe University, Frankfurt am Main, Germany.
Department of Cell Biology & Applied Virology, Fraunhofer Institute for Biomedical Engineering, St. Ingbert, Germany.
Eur J Pharm Biopharm. 2014 Oct;88(2):510-7. doi: 10.1016/j.ejpb.2014.06.008. Epub 2014 Jul 1.
Severe intoxications with organophosphates require the immediate administration of atropine in combination with acetyl cholinesterase (AChE) reactivators such as HI-6. Although this therapy regimen enables the treatment of peripheral symptoms, the blood-brain barrier (BBB) restricts the access of the hydrophilic antidotes to the central nervous system which could lead to a fatal respiratory arrest. Therefore, HI-6-loaded albumin nanoparticles were previously developed to enhance the transport across this barrier and were able to reactivate organophosphate-(OP)-inhibited AChE in an in vitro BBB model. Since HI-6 is known to be moisture-sensitive, the feasibility of freeze-drying of the HI-6-loaded nanoparticles was investigated in the present study using different cryo- and lyoprotectants at different concentrations. Trehalose and sucrose (3%, w/v)-containing formulations were superior to mannitol concerning the physicochemical parameters of the nanoparticles whereas trehalose-containing samples were subject of a prolonged storage stability study at temperatures between -20°C and +40°C for predetermined time intervals. Shelf-life computations of the freeze-dried HI-6 nanoparticle formulations revealed a shelf-life time of 18 months when stored at -20°C. The formulations' efficacy was proven in vitro by reactivation of OP-inhibited AChE after transport over a porcine brain capillary endothelial cell layer model.
重度有机磷中毒需要立即给予阿托品，并联合使用乙酰胆碱酯酶（AChE）复活剂，如HI-6。尽管这种治疗方案能够治疗外周症状，但血脑屏障（BBB）会限制亲水性解毒剂进入中枢神经系统，这可能导致致命的呼吸骤停。因此，此前研发了负载HI-6的白蛋白纳米颗粒，以增强其跨越该屏障的转运能力，并且在体外血脑屏障模型中能够使被有机磷酸酯（OP）抑制的AChE重新活化。由于已知HI-6对湿度敏感，本研究使用不同浓度的不同冷冻保护剂和冻干保护剂，研究了负载HI-6的纳米颗粒冻干的可行性。就纳米颗粒的物理化学参数而言，含海藻糖和蔗糖（3%，w/v）的制剂优于甘露醇，而含海藻糖的样品在-20°C至+40°C的温度下进行了预定时间间隔的长期储存稳定性研究。冻干的HI-6纳米颗粒制剂的保质期计算表明，在-20°C储存时保质期为18个月。通过在猪脑毛细血管内皮细胞层模型上转运后重新活化被OP抑制的AChE，在体外证明了该制剂的有效性。
