Camacho-Hernandez Gisela Andrea, Casiraghi Andrea, Rudin Deborah, Luethi Dino, Ku Therese C, Guthrie Daryl A, Straniero Valentina, Valoti Ermanno, Schütz Gerhard J, Sitte Harald H, Newman Amy Hauck
Medicinal Chemistry Section, Molecular Targets and Medications Discovery Branch, National Institutes of Drug Abuse - Intramural Research Program Baltimore MD 21224 USA
Department of Pharmaceutical Sciences, University of Milan Via Mangiagalli 25 20133 Milan Italy.
RSC Med Chem. 2021 Jun 9;12(7):1174-1186. doi: 10.1039/d1md00072a. eCollection 2021 Jul 21.
The utilization of fluorescent ligands to study the monoamine transporters (MATs) has increased our knowledge of their function and distribution in live cell systems. In this study, we extend SAR for nisoxetine and talopram as parent compounds, to identify high affinity rhodamine-labeled fluorescent probes for the norepinephrine transporter (NET). Nisoxetine-based fluorescent probe demonstrated high binding affinity ( = 43 nM) for NET and an overall selectivity compared to the other transporters for dopamine (DAT; = 1540 nM) and serotonin (SERT; = 785 nM) in competitive radioligand binding assays. Using confocal microscopy, compound was shown to stain both NET and SERT, but not DAT, at low nanomolar concentrations, in transporter-expressing cells.
利用荧光配体研究单胺转运体（MATs）增加了我们对其在活细胞系统中的功能和分布的了解。在本研究中，我们扩展了以尼索西汀和他洛普明为母体化合物的构效关系研究，以鉴定用于去甲肾上腺素转运体（NET）的高亲和力罗丹明标记荧光探针。基于尼索西汀的荧光探针在竞争性放射性配体结合试验中对NET表现出高结合亲和力（Kd = 43 nM），与多巴胺转运体（DAT；Kd = 1540 nM）和5-羟色胺转运体（SERT；Kd = 785 nM）相比具有总体选择性。使用共聚焦显微镜，在低纳摩尔浓度下，化合物在表达转运体的细胞中显示出可同时标记NET和SERT，但不标记DAT。
