摘要
大部分药物需要与细胞膜作用后到达生物靶标,因此,膜亲和力测定是药物发现过程中重要的早期筛选环节。然而,目前仍采用传统的油水分配法,亟需一种新型、简便、准确的膜亲和力测定方法。本研究基于比色原理,优化了一种基于聚联乙炔(polydiacetylene,PDA)囊泡的新型测定模型,通过一系列不同浓度的囊泡溶液和在不同温度及pH反应环境下的实验,筛选出了最优条件。在此基础上,采用盐酸丁卡因、2-甲基咪唑和组胺作为模型药物,测定其膜亲和力常数,并验证了优化后检测模型的批间精密度(相对标准偏差小于5%)。此外,聚联乙炔囊泡具有长达180天的稳定性,证明了该模型的应用潜力。本策略具有操作简便、稳定可靠、高重现性、成本低廉、易于推广等优点,为膜亲和力高通量测定提供了新的工具和新的解决思路。
Most drugs need to interact with cell membrane to reach the biological target,so that membrane affinity assay is an important early screening step in drug discovery.However,at present,the traditional oil-water distribution method is still used,a new,simple and accurate method for membrane affinity assay is urgently needed.In this study,according to the colorimetric principle,a new assay model based on polydiacetylene vesicles was optimized through a series of experiments including different concentrations of vesicle solution,temperature,or pH reaction environment.On this basis,tetracaine hydrochloride,2-methylimidazole and histamine were used as model drugs to measure the membrane affinity constants and verify the between-batch precision of the optimized assay model(relative standard deviation less than 5%).In addition,polydiacetylene vesicles were stable for up to 180 days,demonstrating the potential application of the assay model.This strategy is simple,stable,reliable,with high reproducibility,low cost and easy to promote,which provided a new tool and a new direction for the highthroughput assay of membrane affinity.
作者
聂钗钗
董瑞婷
吴宇彤
吴菁博
张圣
郑枫
丁娅
NIE Chai-chai;DONG Rui-ting;WU Yu-tong;WU Jing-bo;ZHANG Sheng;ZHENG Feng;DING Ya(Department of Pharmaceutical Analysis,College of Pharmacy,China Pharmaceutical University,Nanjing 211112,China)
出处
《药学学报》
CAS
CSCD
北大核心
2023年第8期2503-2511,共9页
Acta Pharmaceutica Sinica
基金
国家自然科学基金资助项目(31870946,32271453)
中国药科大学双一流前瞻基础研究专项(CPU2022QZ12)
中国药科大学2022年大学生创新创业训练计划项目(202210316113).
