Department of Medicinal Chemistry, Key Laboratory of Chemical Biology (MOE), School of Pharmacy, Cheeloo College of Medicine, Shandong University, Jinan 250012, Shandong, China.
Anal Chem. 2022 May 17;94(19):7021-7028. doi: 10.1021/acs.analchem.1c05646. Epub 2022 May 3.
By fusing several environment-sensitive fluorophores to the pharmacophore mirabegron, a series of new fluorescent ligands for β-adrenergic receptors (β-ARs) were produced with a turn-on mechanism and high binding affinity to β-ARs efficiently. Compound with the pyridinium moiety possessed the most favorable combination of properties after systematic comparison and optimization, including high affinity and acceptable cytotoxicity, remarkable fluorescence enhancement (up to 30-fold) upon binding with β-ARs, and feasible visualizing ability of β-ARs in living cells under no-wash conditions. Furthermore, a NanoLuc-based bioluminescence resonance energy transfer (NanoBRET) binding assay based on compound was developed and may be used in high-throughput screening (HTS) in the drug discovery of β-ARs due to its unique fluorescence spectroscopic features. Overall, as the first environment-sensitive fluorescent ligand, molecule could be a useful tool for understanding the pharmacology of β-ARs.
通过将几个环境敏感荧光团融合到药效团米拉贝隆上，我们制备了一系列具有开环机制的新型荧光β-肾上腺素能受体（β-AR）配体，对β-AR 具有高效的结合亲和力。在系统比较和优化后，具有吡啶基部分的化合物 具有最有利的性质组合，包括高亲和力和可接受的细胞毒性、与 β-AR 结合时显著的荧光增强（高达 30 倍），以及在无洗涤条件下在活细胞中可视化 β-AR 的可行性。此外，我们还基于化合物 开发了一种基于 NanoLuc 的生物发光共振能量转移（NanoBRET）结合测定法，由于其独特的荧光光谱特征，该测定法可用于 β-AR 药物发现中的高通量筛选（HTS）。总的来说，作为第一个环境敏感荧光配体，分子 可以成为理解 β-AR 药理学的有用工具。
