中华眼镜蛇神经毒素可溶性微针的制备、表征及其离体皮肤渗透研究  

Preparation and characterization of dissolvable microneedles of Naja atra neurotoxin and their ex vivo skin permeation

作　　者：徐莹莹 周珊珊 温乐 杨柳 裘茂良 潘佳琳 单钰君 郑杭生[1] XU Yingying;ZHOU Shanshan;WEN Le;YANG Liu;QIU Maoliang;PAN Jialin;SHAN Yujun;ZHENG Hangsheng(School of Pharmaceutical Sciences,Zhejiang Chinese Medical University,Hangzhou 311402,China;Zhejiang Business College,Hangzhou 310053,China;Hangzhou 3DSKIN Biotechnology Co.,Ltd.,Hangzhou 310005,China)

机构地区：[1]浙江中医药大学院,杭州311402 [2]浙江商业职技术学院,杭州310053 [3]杭州德肤修生物科技有限公司,浙江杭州310005

出　　处：《中草药》2024年第12期3966-3976,共11页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金项目(82174096);浙江省大学生创新训练项目(202310344055)。

摘　　要：目的筛选多种常用的微针基质辅料,制备具有合适机械强度和柔韧性的载中华眼镜蛇神经毒素(neurotoxin,NT)可溶性微针(dissolvable microneedles,DMN)(DMN-NT),并对其进行体内外评价。方法采用两步真空模板填充法制备不同材料构成的DMN,并对其进行力学特性分析,通过重量法对其吸湿性进行考察,利用正置显微镜对其形态特征进行考察,筛选最佳基质材料后制备DMN-NT,并对微针的载药量、机械性能、在体溶解、在体恢复、体外释放度、离体皮肤渗透与稳定性等进行评价。结果针尖最佳基质材料为20%聚乙烯吡咯烷酮K90(PVP K90)和20%透明质酸(相对分子质量为10000),背衬层最佳基质材料为7.5%羟丙基甲基纤维素(HPMC,E5LV型)和15%聚乙烯醇(PVA-0588)。正置显微镜和体式显微镜下观察发现DMN-NT针体垂直,排列整齐;每片DMN-NT的载药量为(172.22±1.30)μg或(17.38±0.57)μg;力学性能测试结果显示,DMN-NT针尖受力可达(39.88±3.09)N,大于微针穿刺皮肤所需的破坏力0.05 N,表明DMN-NT有较好的机械强度,亚甲基蓝染色实验结果表明微针具有较好的穿刺性;在体溶解实验结果显示,在10 min内DMN-NT大部分溶解;在体恢复实验结果为给药30 min后针孔痕迹几乎不见,表明DMN-NT的生物相容性好,安全性高;体外释放结果表明DMN-NT在pH值为7.4的PBS中8 min的累积释放率达(93.65±4.29)%;离体皮肤渗透结果显示,10 h后微针的累积药物渗透百分率达到(97.99±2.80)%,而药物水溶液中药物几乎没有透过皮肤;DMN-NT稳定性初步评价结果良好。结论制备的DMN可以穿刺皮肤角质层,具有较好的溶解性和生物相容性,可以高效经皮递送大分子药物。Objective To screen various commonly used microneedle matrix excipients and prepare dissolvable microneedles(DMN)with suitable mechanical strength and flexibility for delivering Chinese cobra(Naja atra Laurenti)-derived neurotoxin(NT).Methods The DMNs were prepared by using a two-step vacuum mold-filling method.Their morphological,mechanical and hygroscopic properties were characterized by upright microscope,texture analyzer and gravimetric method respectively to select the optimal matrix material.Then the DMN loaded with neurotoxin(DMN-NT)was prepared with the optimal matrix material,and the drug loading,mechanical properties,in vivo dissolving behavior,in vivo skin recovery,in vitro drug release,ex vivo transdermal permeation and stability of the DMN-NT were evaluated.Results The optimal matrix materials for the needle body were 20%of polyvinylpyrrolidone K90(PVP K90)and 20%of hyaluronic acid(MW 10000),while the backing layer was composed of 7.5%of hydroxypropyl methyl cellulose(HPMC,E5LV)and 15%of polyvinyl alcohol(PVA-0588).Under the upright microscope and the stereomicroscope,the DMN-NT needles were arranged neatly and vertically.Each DMN-NT patch had a drug loading of(172.22±1.30)μg or(17.38±0.57)μg.The mechanical properties evaluation results showed that the pressure applied to the DMN-NT needle tip could reach(39.88±3.09)N,exceeding the force required(0.05 N)for microneedle to penetrate the skin,indicating good mechanical strength.The results of methylene blue staining experiments showed that the DMN-NT had desirable penetration property.In the in vivo dissolving experiment,most of the DMN-NT dissolved within 10 min.The in vivo recovery experiment showed that the trace in skin induced by DMN-NT were almost invisible 30 min after administration,indicating good biocompatibility and high safety.The in vitro release test results showed that the cumulative released percent of DMN-NT in PBS with pH 7.4 reached(93.65±4.29)%in 8 min.The results of the ex vivo skin permeation test showed that the transdermal

关 键 词：中华眼镜蛇 神经毒素 可溶性微针 机械性能 体外释放 经皮药物递送 离体皮肤渗透 

分 类 号：R283.6[医药卫生—中药学]