Strutt Robert, Berlanda Simon F, Dittrich Petra S
Department of Biosystems Science and Engineering, ETH Zürich, Schanzenstrasse 44, 4056 Basel, Switzerland.
Anal Chem. 2025 Aug 19;97(32):17472-17480. doi: 10.1021/acs.analchem.5c02264. Epub 2025 Aug 5.
Membrane transport is fundamental to biological cells and is a major hurdle in the rational design of pharmaceuticals. To measure membrane transport in vitro, most methods focus on simple diffusion of a single analyte across nonbiomimetic interfaces. Membrane engineering has facilitated novel strategies for the reconstitution, characterization, and application of biomimetic membranes. Herein, we define drug mixture analysis, an in vitro, label-free HPLC-MS, droplet interface bilayer (DIB) method, to assess membrane transport of drug mixtures and delineate simultaneous transport mechanisms. We use our method to classify the permeability of drugs from a structurally diverse FDA-approved library. This deep analysis uncovered correlation between determined permeability classifiers and drug properties such as the hydrophobic retention time, hydrogen bond donor count, lipophilicity, and predicted gut absorption. Across higher mimetic membranes, passive transport was quantified under physiologically relevant variables such as pH, temperature, lipid composition, and, in the presence of proteins, the coexistence of facilitated diffusion and simple diffusion. Our results show that DIBs are physiologically relevant interfaces for investigating membrane transport mechanisms relevant to artificial cell systems and drug screening.
膜转运是生物细胞的基础，也是药物合理设计中的一个主要障碍。为了在体外测量膜转运，大多数方法集中于单一分析物在非仿生界面上的简单扩散。膜工程促进了仿生膜重构、表征和应用的新策略。在此，我们定义了药物混合物分析，这是一种体外、无标记的高效液相色谱 - 质谱联用、液滴界面双层（DIB）方法，用于评估药物混合物的膜转运并描绘同时发生的转运机制。我们使用我们的方法对来自结构多样的美国食品药品监督管理局（FDA）批准库中的药物渗透性进行分类。这种深入分析揭示了所确定的渗透性分类器与药物性质之间的相关性，如疏水保留时间、氢键供体数量、亲脂性和预测的肠道吸收。在更高仿生度的膜上，在生理相关变量如pH、温度、脂质组成以及存在蛋白质时，促进扩散和简单扩散共存的情况下，对被动转运进行了量化。我们的结果表明，DIBs是用于研究与人工细胞系统和药物筛选相关的膜转运机制的生理相关界面。
