Design and Synthesis of Fluorescent Methylphenidate Analogues for a FRET-Based Assay of Synapsin III Binding.,ChemMedChem

当前位置： X-MOL 学术 › ChemMedChem › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Design and Synthesis of Fluorescent Methylphenidate Analogues for a FRET-Based Assay of Synapsin III Binding.
ChemMedChem ( IF 3.6 ) Pub Date : 2020-05-26 , DOI: 10.1002/cmdc.202000128
Andrea Casiraghi _{1,

2} , Francesca Longhena ₃ , Valentina Straniero ₁ , Gaia Faustini ₃ , Amy H Newman ₂ , Arianna Bellucci ₃ , Ermanno Valoti ₁

Affiliation

We previously described synapsin III (Syn III) as a synaptic phosphoprotein that controls dopamine release in cooperation with α‐synuclein (aSyn). Moreover, we found that in Parkinson's disease (PD), Syn III also participates in aSyn aggregation and toxicity. Our recent observations point to threo‐methylphenidate (MPH), a monoamine re‐uptake inhibitor that efficiently counteracts the freezing‐gait characteristic of advanced PD, as a ligand for Syn III. We have designed and synthesised two different fluorescently labelled MPH derivatives, one with Rhodamine Red (RHOD) and one with 5‐carboxytetramethylrhodamine (TAMRA), to be used for assessing MPH binding to Syn III by FRET. TAMRA‐MPH exhibited the ideal characteristics to be used as a FRET acceptor, as it was able to enter into the SK‐N‐SH cells and could interact specifically with human green fluorescent protein (GFP)‐tagged Syn III but not with GFP alone. Moreover, the uptake of TAMRA‐MPH and co‐localization with Syn III was also observed in primary mesencephalic neurons. These findings support that MPH is a Syn III ligand and that TAMRA‐conjugated drug molecules might be valuable tools to study drug‐ligand interactions by FRET or to detect Syn III in cytological and histological samples.

中文翻译：

用于基于 FRET 的突触蛋白 III 结合测定的荧光哌甲酯类似物的设计和合成。

我们之前将突触蛋白 III (Syn III) 描述为一种突触磷蛋白，可与 α-突触核蛋白 (aSyn) 配合控制多巴胺的释放。此外，我们发现在帕金森病（PD）中，Syn III也参与aSyn聚集和毒性。我们最近的观察表明，苏型哌甲酯 (MPH) 是一种单胺再摄取抑制剂，可有效抵消晚期 PD 的冻结步态特征，可作为 Syn III 的配体。我们设计并合成了两种不同的荧光标记 MPH 衍生物，一种是罗丹明红 (RHOD)，另一种是 5-羧基四甲基罗丹明 (TAMRA)，用于通过 FRET 评估 MPH 与 Syn III 的结合。 TAMRA-MPH 表现出用作 FRET 受体的理想特性，因为它能够进入 SK-N-SH 细胞，并且可以与人绿色荧光蛋白 (GFP) 标记的 Syn III 特异性相互作用，但不能单独与 GFP 相互作用。此外，在原代中脑神经元中也观察到 TAMRA-MPH 的摄取以及与 Syn III 的共定位。这些发现支持 MPH 是 Syn III 配体，并且 TAMRA 缀合的药物分子可能是通过 FRET 研究药物-配体相互作用或检测细胞学和组织学样品中 Syn III 的有价值的工具。

更新日期：2020-07-20

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11