MPX-004 and MPX-007: New Pharmacological Tools to Study the Physiology of NMDA Receptors Containing the GluN2A Subunit.
MPX-004 and MPX-007: New Pharmacological Tools to Study the Physiology of NMDA Receptors Containing the GluN2A Subunit.
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DOI：
10.1371/journal.pone.0148129
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Menniti FS
中科院分区：
文献类型：
--
作者：
Volkmann RA;Fanger CM;Anderson DR;Sirivolu VR;Paschetto K;Gordon E;Virginio C;Gleyzes M;Buisson B;Steidl E;Mierau SB;Fagiolini M;Menniti FS
GluN 2A是哺乳动物CNS中含量最丰富的GluN 2 NMDA受体亚单位。生理学和遗传学证据表明，含GluN 2A的受体与自闭症、精神分裂症、儿童癫痫和Rett综合征等神经发育障碍的易感性有关。然而，GluN 2A选择性药理学探针，以探索靶向这些受体的治疗潜力一直缺乏。在此，我们公开了一系列新的含吡嗪的GluN 2A拮抗剂，例如MPX-004（5-（3-氯-4-氟苯基）磺酰氨基）甲基）-N-（（2-甲基噻唑-5-基）甲基）吡嗪-2-甲酰胺）和MPX-007（5-（3-氟-4-氟苯基）磺酰氨基）甲基）-N-（（2-甲基噻唑-5-基）甲基）甲基吡嗪-2-甲酰胺）。MPX-004和MPX-007抑制HEK细胞中表达的含GluN 2A的NMDA受体，IC 50分别为79 nM和27 nM。相反，在完全抑制GluN 2A活性的浓度下，这些化合物在类似的基于HEK细胞的测定中对GluN 2B或GluN 2D受体介导的应答没有抑制作用。在表达GluN 2A-D受体亚型的非洲爪蟾卵母细胞的电生理学测定中证实了效力和选择性。MPX-004和MPX-007的最大浓度抑制原代培养的大鼠锥体神经元中约30%的全细胞电流，并且MPX-004抑制大鼠海马切片中约60%的总NMDA受体介导的EPSP。通过MPX-004对来自GRIN 2A敲除小鼠的皮质切片中的NMDA受体介导的突触电流没有抑制作用的发现证实了天然受体处的GluN 2A选择性。因此，MPX-004和MPX-007提供了高度选择性的药理学工具，以探测GluN 2A生理学和参与神经精神和发育障碍。
GluN2A is the most abundant of the GluN2 NMDA receptor subunits in the mammalian CNS. Physiological and genetic evidence implicate GluN2A-containing receptors in susceptibility to autism, schizophrenia, childhood epilepsy and neurodevelopmental disorders such as Rett Syndrome. However, GluN2A-selective pharmacological probes to explore the therapeutic potential of targeting these receptors have been lacking. Here we disclose a novel series of pyrazine-containing GluN2A antagonists exemplified by MPX-004 (5-(((3-chloro-4-fluorophenyl)sulfonamido)methyl)-N-((2-methylthiazol-5-yl)methyl)pyrazine-2-carboxamide) and MPX-007 (5-(((3-fluoro-4-fluorophenyl)sulfonamido)methyl)-N-((2-methylthiazol-5-yl)methyl)methylpyrazine-2-carboxamide). MPX-004 and MPX-007 inhibit GluN2A-containing NMDA receptors expressed in HEK cells with IC50s of 79 nM and 27 nM, respectively. In contrast, at concentrations that completely inhibited GluN2A activity these compounds have no inhibitory effect on GluN2B or GluN2D receptor-mediated responses in similar HEK cell-based assays. Potency and selectivity were confirmed in electrophysiology assays in Xenopus oocytes expressing GluN2A-D receptor subtypes. Maximal concentrations of MPX-004 and MPX-007 inhibited ~30% of the whole-cell current in rat pyramidal neurons in primary culture and MPX-004 inhibited ~60% of the total NMDA receptor-mediated EPSP in rat hippocampal slices. GluN2A-selectivity at native receptors was confirmed by the finding that MPX-004 had no inhibitory effect on NMDA receptor mediated synaptic currents in cortical slices from GRIN2A knock out mice. Thus, MPX-004 and MPX-007 offer highly selective pharmacological tools to probe GluN2A physiology and involvement in neuropsychiatric and developmental disorders.