血脑屏障渗透模型PAMPA的构建及其条件优化

Model Establishment and Optimized Conditions of PAMPA for Blood-Brain Barrier

平行人工膜渗透模型(parallel artificial membrane permeation assay,PAMPA)主要以人工磷脂作为生物膜来模拟药物跨膜的屏障,能测定通过被动扩散途径跨过血脑屏障的药物,该模型操作简单,成本低。以实验室合成的化合物为研究对象,在MultiScreen~■ 96-well板上铺一层牛脑磷脂的十二烷溶液作为PAMPA模型,并通过高效液相色谱(HPLC)测定化合物的血脑屏障渗透率。实验中发现亲脂性化合物水溶性较差,需要添加助溶剂以帮助溶解,因此考察了4种助溶剂(DMSO、乙腈、乙醇、正丙醇)对化合物溶解度的影响及助溶剂含量对磷脂膜完整性的影响,最终选择了浓度为10%的正丙醇做为助溶剂来进行渗透实验,测得了各个化合物的渗透率,并筛选出了渗透率最好的化合物,为将来的药物开发提供指导。

Parallel artificial membrane permeation assay(PAMPA) mainly uses artificial phospholipids as biofilm to simulate the barrier of drug transmembrane, and can measure drugs that cross the blood-brain barrier through passive diffusion. The model is simple in operation and low in cost. In this study, a laboratory-synthesized compound was used as a research object. A layer of bovine brain phospholipid dodecane solution was applied to the MultiScreen ■ 96-well plate as a PAMPA model, and the Blood-Brain Barrier Permeability of the compound was determined by high performance liquid chromatography(HPLC). In the experiment, it was found that the lipophilic of four cosolvents(DMSO, acetonitrile, ethanol, n-propanol) on the solubility of the compound and compound was poorly water-soluble. To solve this problem, this experiment investigated the effects of the concentration of the cosolvent on the integrity of the phospholipid membrane. The effect was chosen to select the best cosolvent for the penetration test of the Blood-Brain Barrier penetration model PAMPA.