HI-6人体血清白蛋白纳米粒的制备及其对梭曼中毒小鼠脑AChE重活化作用评价

迅速恢复中毒乙酰胆碱酯酶(acetylcholinesterase,AChE)活性是神经性毒剂中毒救治最关键的环节.血脑屏障的存在限制了具有重活化作用的抗毒药物以有效治疗剂量进入中枢.本研究以已知重活化作用最强的重活化剂酰胺磷定(HI-6)为目标药物,通过去溶剂化法制备了人体血清白蛋白纳米粒,通过静电作用将HI-6分子装载在纳米粒表面,合成装载HI-6的人体血清白蛋白纳米粒;在斑马鱼及神经性毒剂梭曼染毒小鼠上,对药物穿透血脑屏障能力及中枢中毒AChE重活化作用进行了评价.结果表明,装载HI-6人体血清白蛋白纳米粒在物理表征上符合纳米药物基本特征;相对于自由HI-6,HI-6人体血清白蛋白纳米粒能够透过血脑屏障,并将中枢中毒AChE活性提升2倍以上,表明人体血清白蛋白纳米粒成功携带HI-6分子进入中枢.本文建立了一种无毒、高效、小尺寸中枢靶向性纳米粒的制备方法,基于该方法制备的纳米重活化剂,在脑内可有效释放目标药物并迅速发挥解毒作用.

Preparation of HI-6 absorptive human serum albumin nanoparticles and evaluation of its reactivation efficacy on inhibited brain AChE in soman-intoxicated mice

It was essential to rapidly reactivate and restore the function of the inhibited acetylcholinesterase（AChE） in the treatment of nerve agents poisoning. However, the blood-brain barrier （BBB） restricts the rapid transport of reactivators from the blood into the brain in therapeutically relevant concentrations. In this study, human serum albumin nanoparticles（HSA NPs） were prepared via desolvent method; HI-6, the known reactivator with higher reactivating efficiency were bound to HSA NPs through electrostatic interaction; at last, on zebrafish BBB model and soman-intoxicated mice, the permeability on BBB and the reactivation on inhibited brain AChE of nanoparticulate oxime formulations were evaluated. All characterization data revealed that HSA NPs loaded with HI-6 had met the basic demand for nanodrug therapy. Compared with free HI-6, HSA NPs loaded HI-6 could cross the BBB and improve the reactivating rates two times, suggesting HSA NPs could carry HI-6 into CNS successfully. In brief, we improved and established a method to prepare the brain-targeted nanoparticles with small-size, low-toxicity and high-efficiency, the targeted drug based on this method could efficiently release and rapidly antagonize the nerve agents poisoning.