Geiger Robert M, Rahman Shekh M, Rashid Roni Md Shadiqur, Sullenberger Catherine, Mistry Sabyasachy, Shea Katherine, Tariq Isra, Ismaiel Omnia A, Matta Murali K, Hyland Paula L, Berdichevski Sasha, Ribeiro Alexandre J S, Blinova Ksenia, Jiang Wenlei, Walenga Ross L, Newman Bryan, Volpe Donna A, Ford Kevin A
Division of Applied Regulatory Science, Office of Clinical Pharmacology, Office of Translational Sciences, Center for Drug Evaluation and Research, United States Food and Drug Administration, Silver Spring, MD, United States.
Emulate, Inc., Boston, MA, United States.
Front Pharmacol. 2025 Jul 17;16:1621775. doi: 10.3389/fphar.2025.1621775. eCollection 2025.
There is a need to reliably predict the permeability of inhaled compounds during the development of new and generic drugs. A small airway microphysiological system (MPS) that can recapitulate the pulmonary air-liquid interface (ALI) with primary epithelial and vascular endothelial cell layers may provide a more physiologically relevant environment for measuring drug permeability than simpler two-dimensional cell culture platforms. Therefore, we evaluated the use of a small airway MPS to measure the permeability of inhaled drugs.
Primary human lung epithelial cells were seeded onto the top channel of the chip and cultured for 14 days at ALI to promote monolayer differentiation, followed by addition of endothelial cells into the bottom channel. Due to the non-specific binding properties of polydimethylsiloxane (PDMS), a drug absorption study was conducted to quantify non-specific binding to the material. Drug permeability was evaluated by passing each compound (10 µM) through the top channel and measuring the amount of drug that permeated into the bottom channel over the time course of 30, 60, 120, and 180 min.
Confocal micrographs demonstrated the presence of tight junctions along with basal, goblet, and ciliated cells in the top channel and attachment of endothelial cells in the bottom channel. Insignificant nonspecific binding to the MPS was observed with albuterol sulfate, formoterol fumarate, and olodaterol hydrochloride (HCl), while fluticasone furoate showed significant nonspecific binding as only 6%-44% of the drug was recovered at 30 and 120 min, respectively. As a result, fluticasone furoate was excluded from further analysis. Permeability studies estimated an apparent permeability (P) of 1.02 × 10 cm/s for albuterol sulfate, 0.0813 × 10 cm/s for olodaterol HCl, and 2.44 × 10 cm/s for formoterol fumarate.
Taken together, the small airway MPS recapitulated relevant cell types and many morphological features in the lung. The apparent permeabilities measured indicated that albuterol sulfate and formoterol fumarate would be categorized as highly permeable, while olodaterol HCl would be categorized as a low permeable drug.
在新型药物和仿制药的研发过程中，需要可靠地预测吸入性化合物的渗透性。一种能够通过原代上皮细胞层和血管内皮细胞层重现肺气液界面（ALI）的小气道微生理系统（MPS），可能比更简单的二维细胞培养平台提供更具生理相关性的环境来测量药物渗透性。因此，我们评估了使用小气道MPS来测量吸入药物的渗透性。
将原代人肺上皮细胞接种到芯片的顶部通道，并在ALI条件下培养14天以促进单层分化，随后将内皮细胞添加到底部通道。由于聚二甲基硅氧烷（PDMS）的非特异性结合特性，进行了药物吸收研究以量化与该材料的非特异性结合。通过使每种化合物（10μM）通过顶部通道，并在30、60、120和180分钟的时间过程中测量渗透到底部通道的药物量来评估药物渗透性。
共聚焦显微镜图像显示顶部通道中存在紧密连接以及基底细胞、杯状细胞和纤毛细胞，底部通道中有内皮细胞附着。硫酸沙丁胺醇、富马酸福莫特罗和盐酸奥洛他定对MPS的非特异性结合不显著，而糠酸氟替卡松显示出显著的非特异性结合，因为在30分钟和120分钟时分别仅回收了6%-44%的药物。因此，糠酸氟替卡松被排除在进一步分析之外。渗透性研究估计硫酸沙丁胺醇的表观渗透率（P）为1.02×10 cm/s，盐酸奥洛他定的表观渗透率为0.0813×10 cm/s，富马酸福莫特罗的表观渗透率为^{ }2.44×10 cm/s。
总体而言，小气道MPS重现了肺中的相关细胞类型和许多形态特征。测得的表观渗透率表明，硫酸沙丁胺醇和富马酸福莫特罗将被归类为高渗透性药物，而盐酸奥洛他定将被归类为低渗透性药物。
