Identification and characterization of ML352: a novel, noncompetitive inhibitor of the presynaptic choline transporter.
Identification and characterization of ML352: a novel, noncompetitive inhibitor of the presynaptic choline transporter.
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DOI：
10.1021/cn5001809
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发表时间：
2015-03-18
影响因子：
5
通讯作者：
Blakely, Randy D.
中科院分区：
文献类型：
--
作者：
Ennis, Elizabeth A.;Wright, Jane;Retzlaff, Cassandra L.;McManus, Owen B.;Lin, Zhinong;Huang, Xiaofang;Wu, Men;Li, Min;Daniels, J. Scott;Lindsley, Craig W.;Hopkins, Corey R.;Blakely, Randy D.
高亲和力胆碱转运体（CHT）是乙酰胆碱（ACh）合成的限速决定因子，然而该转运体仍然是用于检测和操纵突触胆碱能信号的大部分未开发的靶标。为了扩大CHT药理学，我们追求高通量筛选新的CHT靶向小分子的基础上，转运介导的胆碱转运的产电特性。在这项工作中，我们确定了五个新的，结构类的CHT特异性抑制剂。这些类别之一的化学多样化和功能分析将ML 352鉴定为高亲和力（Ki = 92 nM）和选择性CHT抑制剂。在转染细胞和神经末梢制剂中完全拮抗CHT的浓度下，ML 352对乙酰胆碱酯酶（AChE）或胆碱乙酰转移酶（ChAT）无抑制作用，对多巴胺、5-羟色胺和去甲肾上腺素转运蛋白以及许多受体和离子通道也缺乏活性。ML 352在完整细胞和突触体中表现出非竞争性胆碱摄取抑制，并在膜测定中降低了半胆碱-3（HC-3）结合位点的表观密度，表明了变构转运蛋白相互作用。药代动力学研究显示体外代谢有限，CNS渗透显著，具有预测快速清除的特征。ML 352代表了一种新的，有效的，和具体的工具，用于操纵CHT，提供了一个可能的平台，开发胆碱能成像和治疗剂。
The high-affinity choline transporter (CHT) is the rate-limiting determinant of acetylcholine (ACh) synthesis, yet the transporter remains a largely undeveloped target for the detection and manipulation of synaptic cholinergic signaling. To expand CHT pharmacology, we pursued a high-throughput screen for novel CHT-targeted small molecules based on the electrogenic properties of transporter-mediated choline transport. In this effort, we identified five novel, structural classes of CHT-specific inhibitors. Chemical diversification and functional analysis of one of these classes identified ML352 as a high-affinity (Ki = 92 nM) and selective CHT inhibitor. At concentrations that fully antagonized CHT in transfected cells and nerve terminal preparations, ML352 exhibited no inhibition of acetylcholinesterase (AChE) or cholineacetyltransferase (ChAT) and also lacked activity at dopamine, serotonin, and norepinephrine transporters, as well as many receptors and ion channels. ML352 exhibited noncompetitive choline uptake inhibition in intact cells and synaptosomes and reduced the apparent density of hemicholinium-3 (HC-3) binding sites in membrane assays, suggesting allosteric transporter interactions. Pharmacokinetic studies revealed limited in vitro metabolism and significant CNS penetration, with features predicting rapid clearance. ML352 represents a novel, potent, and specific tool for the manipulation of CHT, providing a possible platform for the development of cholinergic imaging and therapeutic agents.
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影响因子：
2.7
作者：
Crooks, PA;Ayers, JT;Dwoskin, LP
通讯作者：
Dwoskin, LP
DOI：
10.1152/ajpheart.00170.2010
发表时间：
2010-09-01
影响因子：
4.8
作者：
English, Brett A.;Appalsamy, Martin;Blakely, Randy D.
通讯作者：
Blakely, Randy D.
DOI：
10.1016/0005-2736(73)90508-7
发表时间：
1973-01-01
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
HAGA, T;NODA, H
通讯作者：
NODA, H
影响因子：
2.5
作者：
Hahn, M. K.;Blackford, J. U.;Shelton, R.
通讯作者：
Shelton, R.
DOI：
10.1038/npp.2010.104
发表时间：
2011-01
期刊：
Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology
影响因子：
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作者：
通讯作者：
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