Li Jing, Wang Bu, Juba Brian M, Vazquez Michael, Kortum Steve W, Pierce Betsy S, Pacheco Michael, Roberts Lee, Strohbach Joseph W, Jones Lyn H, Hett Erik, Thorarensen Atli, Telliez Jean-Baptiste, Sharei Armon, Bunnage Mark, Gilbert Jonathan Brian
Medicine Design, Pfizer, Inc. , 1 Portland Street, Cambridge, Massachusetts 02139, United States.
SQZ Biotechnologies , 134 Coolidge Avenue, Watertown, Massachusetts 02472, United States.
ACS Chem Biol. 2017 Dec 15;12(12):2970-2974. doi: 10.1021/acschembio.7b00683. Epub 2017 Nov 13.
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.
生化筛选是新靶点潜在药物发现的主要来源。然而，在小分子先导化合物优化过程中，具有优异生化活性的化合物可能表现出较差的细胞活性，使得构效关系难以解读。这可能是由于分子的膜通透性低，导致细胞内药物浓度不足。细胞挤压平台通过机械破坏细胞膜来提高通透性，而不管化合物的结构如何，这可以克服通透性限制，弥合生化研究和细胞研究之间的差距。在本研究中，我们表明，使用细胞挤压技术可将通透性差的 Janus 激酶（JAK）抑制剂递送至原代细胞中，抑制高达 90%的 JAK 信号通路，而 JAK 抑制剂在有或无电穿孔的情况下孵育均无显著效果。我们相信，这种强大的细胞内递送方法能够实现更有效的先导化合物优化，并加深我们对小分子和功能探针与靶点相互作用的理解。
