Microfluidic-Enabled Intracellular Delivery of Membrane Impermeable Inhibitors to Study Target Engagement in Human Primary Cells

ACS Chemical Biology
Jing LiJonathan Brian Gilbert

Abstract

Biochemical screening is a major source of lead generation for novel targets. However, during the process of small molecule lead optimization, compounds with excellent biochemical activity may show poor cellular potency, making structure-activity relationships difficult to decipher. This may be due to low membrane permeability of the molecule, resulting in insufficient intracellular drug concentration. The Cell Squeeze platform increases permeability regardless of compound structure by mechanically disrupting the membrane, which can overcome permeability limitations and bridge the gap between biochemical and cellular studies. In this study, we show that poorly permeable Janus kinase (JAK) inhibitors are delivered into primary cells using Cell Squeeze, inhibiting up to 90% of the JAK pathway, while incubation of JAK inhibitors with or without electroporation had no significant effect. We believe this robust intracellular delivery approach could enable more effective lead optimization and deepen our understanding of target engagement by small molecules and functional probes.

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Citations

Nov 2, 2018·Proceedings of the National Academy of Sciences of the United States of America·Tia DiTommasoArmon Sharei
Jan 10, 2019·Lab on a Chip·Chang-Koo YunYong-Soo Choi
Jan 11, 2020·Frontiers in Chemistry·Daniel ConoleEdward W Tate
Feb 18, 2021·Advanced Materials·Dorsa Morshedi RadMajid Ebrahimi Warkiani
Apr 17, 2021·Advanced Materials·Arun R K KumarAndy Tay
Jul 3, 2021·Materials·Noshad PeyravianMasoud Mozafari
Jul 20, 2021·Chemical Reviews·Yuebao ZhangYizhou Dong
Jul 17, 2021·Advanced Materials·Pengchao ZhangLidong Qin
Aug 10, 2021·Biomicrofluidics·Mehdi NikfarYaling Liu
Jul 28, 2018·Chemical Reviews·Martin P StewartKlavs F Jensen

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