目的: 基于平行人工膜渗透分析法(parallel artificial membrane permeability analysis,PAMPA),对化合物跨血脑屏障(blood-brain barrier,BBB)仿生系统进行优化。以氯丙嗪、氯氮平、马普替林及维拉帕米等10种化合物进行渗透性考察,建立1种快速、简便的化合物BBB渗透性考察方法。方法: 使用μFlux型化合物渗透性测定仪,以人工仿生膜为载体,将BBB涂膜液涂于仿生膜上制备人工渗透膜。对该平行人工膜模型进行方法学验证,考察其线性、重现性及重复性。以人工渗透膜模拟人的BBB,考察化合物经人工渗透膜的有效渗透速率(Pe),并与文献报道的通过96孔板测得的跨膜渗透速率(Pm)进行比较,验证人工渗透膜的准确性。结果: 该方法测得的各化合物均具有良好的线性关系(r＞0.999),不同日期测定维拉帕米Pe值一致(P＞0.05),该方法具有良好的重现性与重复性。对各化合物实验结果经过统计学分析,以平行人工膜测定的Pe与96孔板测定的Pm在统计学上具有一定的一致性(选择系数为1.003 63)。结论: 基于化合物跨BBB仿生系统优化PAMPA法能测定化合物跨过BBB渗透性,可为创新化合物跨BBB研究提供体外预测。
Objective: To optimize the bionic system for compound trans blood-brain barrier (BBB) based on parallel artificial membrane permeation analysis (PAMPA). The permeability of ten compounds including chlorpromazine, clozapine, maprotiline and verapamil was investigated to establish a quick and simple method for the investigation of compound BBB permeability. Methods: A μFlux compound permeability tester was used with the artificial biomimetic membrane as carrier,and the artificial permeable membrane was prepared by coating the BBB coating solution on the biomimetic membrane. The parallel artificial membrane model was verified by methodology, the linearity, reproducibility and repeatability were investigated. The artificial permeable membrane was used to simulate the human BBB, and the permeability (Pe) of the compound through the artificial permeable membrane was investigated and compared with the transmembrane permeability (Pm) measured by the 96-well plate reported in the literature to verify the accuracy of the artificial permeable membrane. Results: There was a good linear relationship in the standard curves of each compound which measured by this method(r>0.999), and the Pe values of verapamil measured on different days were consistent (P>0.05). This method had good reproducibility and repeatability. After statistical analysis of the experimental results of each compound, Pe measured by parallel artificial membrane and Pm measured by 96-well plate were statistically consistent (selection coefficient=1.003 63). Conclusion: The PAMPA bio-simulating system could measure the permeability of compounds crossing BBB, which can provide in vitro prediction for innovative compounds crossing BBB research.
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