Jones Lyn H, Xu Hua, Fadeyi Olugbeminiyi O
Jnana Therapeutics, Boston, MA, United States.
Pfizer Inc, Medicine Design, Cambridge, MA, United States.
Methods Enzymol. 2019;622:201-220. doi: 10.1016/bs.mie.2019.02.010. Epub 2019 Mar 9.
Phenotypic screening in disease-relevant models identifies small molecule hits with desirable efficacy but often with unknown modes of action. Target identification and validation are integral to successful biomedical research. Technologies are required to validate the biological target (or targets) through which a pharmacological agent is proposed to exert its effects. This work details the rational structure-based design, synthetic preparation and cell-based application of a clickable sulfonyl fluoride chemical probe to directly report on the mechanism of a series of compounds previously discovered in a reporter gene assay. Quantification of drug-target occupancy in living human primary cells enabled a deeper understanding of the molecular pharmacology of the chemotype. The technology described herein should be of broad interest to those involved in chemical biology research and the drug discovery endeavor.
在疾病相关模型中的表型筛选可识别出具有理想疗效但作用机制往往未知的小分子活性物质。靶点识别与验证是成功开展生物医学研究不可或缺的环节。需要利用各种技术来验证一种药物制剂据信是通过何种生物靶点发挥作用的。这项工作详细介绍了一种可点击的磺酰氟化学探针基于合理结构的设计、合成制备及其在细胞层面的应用，以直接揭示先前在报告基因检测中发现的一系列化合物的作用机制。对活的人类原代细胞中药物-靶点占有率的定量分析，有助于更深入地了解该化学类型的分子药理学。本文所述技术应该会引起化学生物学研究和药物研发领域相关人员的广泛兴趣。
