Simultaneous Multiple MS Binding Assays for the Dopamine, Norepinephrine, and Serotonin Transporters,ChemMedChem

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Simultaneous Multiple MS Binding Assays for the Dopamine, Norepinephrine, and Serotonin Transporters
ChemMedChem ( IF 3.6 ) Pub Date : 2018-02-16 , DOI: 10.1002/cmdc.201700737
Patrick Neiens ₁ , Angela De Simone ₂ , Georg Höfner ₁ , Klaus T. Wanner ₁

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In this work, we present label‐free, mass‐spectrometry‐based binding assays (MS Binding Assays), targeting the human dopamine, norepinephrine, and serotonin transporters (hDAT, hNET, and hSERT) in simultaneous binding experiments. Using a validated LC–ESI‐MS/MS method for quantification of the selective dopamine transporter inhibitor (R,R)‐4‐(2‐benzhydryloxyethyl)‐1‐(4‐fluorobenzyl)piperidin‐3‐ol ((R,R)‐D‐84), the selective norepinephrine transporter inhibitor (S,S)‐reboxetine, and the selective serotonin reuptake inhibitor (S)‐citalopram, binding affinities at the three monoamine transporters could be characterized simultaneously in a single binding experiment. The performed simultaneous saturation and competition experiments yielded results that are in good accordance with those determined in MS Binding Assays addressing the monoamine transporters individually. The results obtained from this study underscore the potential of MS Binding Assays for simultaneous affinity determination at different targets, which is difficult to accomplish with conventional radioligand binding assays.

中文翻译：

多巴胺，去甲肾上腺素和5-羟色胺转运蛋白的同时多重MS结合测定

在这项工作中，我们在同时结合实验中针对人多巴胺，去甲肾上腺素和血清素转运蛋白（hDAT，hNET和hSERT），提出了基于质谱的无标记结合测定（MS结合测定）。使用经过验证的LC-ESI-MS / MS方法对选择性多巴胺转运蛋白抑制剂（R，R）-4-（2-苯甲酰氧基乙基）-1-（4-氟苄基）哌啶-3-醇（（R，R）-D-84），选择性去甲肾上腺素转运蛋白抑制剂（S，S）-瑞波西汀和选择性5-羟色胺再摄取抑制剂（S）-西酞普兰，可以在一个结合实验中同时表征三个单胺转运蛋白的结合亲和力。进行的同时饱和和竞争实验产生的结果与在MS结合测定法中确定的单独处理单胺转运蛋白的结果非常吻合。从这项研究中获得的结果强调了在不同靶点同时测定亲和力的MS结合测定的潜力，这是常规放射配体结合测定难以实现的。

更新日期：2018-02-16

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