转铁蛋白受体单克隆抗体OX26修饰的眼镜蛇神经毒素脂质体的制备及其对小鼠的镇痛作用研究

Preparation of α-cobrotoxin Loaded Liposome Modified with Transferrin Receptor Monoclonal Antibody OX26 and Its Analgesic Effects on Mice

目的：制备转铁蛋白受体单克隆抗体OX26修饰的眼镜蛇神经毒素（αCT）脂质体（OX26-αCT-LP）,考察其理化性质、体外释放行为,并研究其镇痛作用。方法：采用薄膜分散-挤出法制备αCT-LP,巯基化后的OX26再与脂质体相连制得OX26-αCT-LP。考察所制脂质体的形态、粒径、Zeta电位;采用透析袋法考察OX26-αCT-LP和αCT-LP在p H 7.4的磷酸盐缓冲液中60 h内的释放特性;采用小鼠热板法考察鼻黏膜滴注生理盐水、αCT原料药、αCT-LP和OX26-αCT-LP对小鼠舔后足反应潜伏期的影响。结果：所制OX26-αCT-LP形态呈球形,均匀圆整,平均粒径为（106.3±5.36）nm,Zeta电位为（-23.17±1.14）m V,药物包封率为（56.82±1.18）%;OX26-αCT-LP和αCT-LP的体外释药行为均符合Weibull方程（r=0.986 7、0.981 0）;与生理盐水比较,αCT-LP和OX26-αCT-LP给药后60~480 min内能明显延长小鼠舔后足反应的潜伏期（P〈0.01或P〈0.05）,且OX26-αCT-LP效果强于αCT-LP（P〈0.01）。结论：成功制得对小鼠有一定镇痛作用的OX26-αCT-LP。

OBJECTIVE： To prepare the a-cobrotoxin （αCT） loaded liposome modified with transferrin receptor monoclonal antibody OX26 （OX26-αCT-LP）, detect the physicochemical properties and release feature in vitro and study the analgesic effects. METHODS： Film dispersion-extrusion was conducted to prepare the αCT-LP and OX26-αCT-LP was prepared by thiolated OX26 and liposomes. The morphology, particle size and Zeta potential were determined; dialysis bag method was used to determine the release characteristics of OX26-αCT-LP and aCT-LP in pH 7.4 phosphate buffer solution within 60 h; mice hot plate method was used to determine the effects of infusion saline of nasal mucosa, αCT raw material medicine, aCT-LP and OX26-αCT-LP on the in- cubation period of the response of mice licking hind foot. RESULTS： The prepared OX26-αCT-LP was spherical, uniform and com- plete, with the average particle size of （106.3 ± 5.36） nm, the Zeta potential of （-23.17 ± 1.14） mV and the encapsulation efficiency of （56.82 ± 1.18）%. The releases in vitro of OX26-αCT-LP and aCT-LP were all conformed to Weibull equation （r=0.986 7, 0.981 0）. Compared with normal saline, αCT-LP and OX26-αCT-LP administration could obviously prolong the incubation period of the response of mice licking hind foot within 60-480 min （P〈0.01 or P〈0.05） and OX26-αCT-LP was better than αCT-LP （P〈 0.01）. CONCLUSIONS： OX26-αCT-LP that has analgesic effect on mice is successfully prepared.