Synthesis and characterization of the first fluorescent antagonists for human 5-HT4 receptors
Synthesis and characterization of the first fluorescent antagonists for human 5-HT4 receptors
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DOI：
10.1021/jm0307887
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发表时间：
2003-06-19
影响因子：
7.3
通讯作者：
Sicsic, S
中科院分区：
文献类型：
--
作者：
Berque-Bestel, I;Soulier, JL;Sicsic, S
基于ML 10302 1（一种有效的5-HT 4受体激动剂）和哌嗪类似物2合成了人5-HT 4受体的荧光拮抗剂。这些分子通过烷基链衍生有三个荧光部分，丹磺酰基、萘酰亚胺和NBD（7-硝基苯-2-氧杂-1，3-二唑-4-基）。在最近克隆的在C6神经胶质细胞中稳定表达的人5-HT 4（e）受体亚型上，以[H-3] GR 113808作为放射性配体，在结合测定中评价合成的分子。亲和力值取决于基础结构以及烷基链长度。基于ML 10302的衍生物是比基于哌嗪类似物的衍生物更有效的配体。对于基于ML 10302的配体，分别通过一个碳原子17 a和32的链长连接的丹磺酰基和NBD衍生物导致接近ML 10302的亲和力。最有效的化合物17 a、28和32在相同的细胞系统中以纳摩尔K-b值产生对5-HT刺激的环AMP合成的抑制。更具体地研究了17 a、28和32的荧光性质。使用h5-HT 4（e）受体转染的C6神经胶质细胞膜和整个细胞显示荧光配体28与h5-HT 4（e）受体的相互作用。配体28也用于荧光显微镜实验，以标记h5-HT 4（e）受体转染的C6神经胶质细胞，并使用共聚焦显微镜更精确地确定这些受体的亚细胞定位。
Fluorescent antagonists for human 5-HT4 receptors were synthesized based on ML10302 1, a potent 5-HT4 receptor agonist and on piperazine analogue 2. These molecules were derived with three fluorescent moieties, dansyl, naphthalimide, and NBD (7-nitrobenz-2-oxa-1,3-diazol-4-yl), through alkyl chains. The synthesized molecules were evaluated in binding assays on the recently cloned human 5-HT4(e) receptor isoform stably expressed in C6 glial cells with [H-3]GR113808 as the radioligand. The affinity values depended upon the basal structure together with the alkyl chain length. The derivatives based on ML10302 were more potent ligands than the derivatives based on piperazine analogue. For ML10302-based ligands, dansyl and NBD derivatives attached through a chain length of one carbon atom 17a and 32, respectively, led to affinities close to the affinity of ML10302. The most potent compounds 17a, 28, and 32 produced an inhibition of the 5-HT stimulated cyclic AMP synthesis in the same cellular system with nanomolar K-b values. Fluorescent properties of 17a, 28, and 32 were more particularly studied. Interactions of the fluorescent ligand 28 with the h5-HT4(e) receptor were indicated using h5-HT4(e) receptor transfected C6 glial cell membranes and entire cells. Ligand 28 was also used in fluorescence microscopy experiments in order to label h5-HT4(e) receptor transfected C6 glial cells, and subcellular localization of these receptors was more precisely determined using confocal microscopy.