Novel human D-amino acid oxidase inhibitors stabilize an active-site lid-open conformation.
Novel human D-amino acid oxidase inhibitors stabilize an active-site lid-open conformation.
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DOI：
10.1042/bsr20140071
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发表时间：
2014-08-11
影响因子：
4
通讯作者：
Large TH
中科院分区：
文献类型：
--
作者：
Terry-Lorenzo RT;Chun LE;Brown SP;Heffernan ML;Fang QK;Orsini MA;Pollegioni L;Hardy LW;Spear KL;Large TH
NMDAR（N-甲基-D-天冬氨酸受体）是突触可塑性和学习记忆的中枢调节因子。hDAAO（人D-氨基酸氧化酶）通过降解NMDAR共激动剂D-丝氨酸间接降低NMDAR活性。由于NMDAR功能减退被认为是精神分裂症的基础缺陷，因此hDAAO抑制剂具有作为精神分裂症和其他神经系统疾病的治疗的潜力。在这里，我们试图鉴定抑制hDAAO活性的新型化学物质。我们使用计算工具来设计一个集中的，可购买的化合物库。在筛选该文库的hDAAO抑制后，我们鉴定了结构新颖的化合物“化合物2”[3-（7-羟基-2-氧代-4-苯基-2H-色烯-6-基）丙酸]，其显示出低nM hDAAO抑制效力（Ki=7 nM）。尽管预期文库富集对D-丝氨酸和FAD都具有竞争性的化合物，但化合物2实际上是FAD非竞争性的，非常像典型的hDAAO抑制剂如苯甲酸。化合物2和类似物独立地与hDAAO共结晶。这些化合物稳定了hDAAO的新构象，其中活性位点盖处于开放位置。这些结果证实了先前关于哺乳动物D-氨基酸氧化酶的活性位点盖子灵活性的假设，并可能有助于下一代hDAAO抑制剂的设计。计算工具与实验测试相结合，用于识别hDAAO（人D-氨基酸氧化酶）的新型抑制剂，hDAAO是一种与精神分裂症和神经性疼痛相关的酶。这些抑制剂稳定了hDAAO构象，酶的活性位点盖子打开。
The NMDAR (N-methyl-D-aspartate receptor) is a central regulator of synaptic plasticity and learning and memory. hDAAO (human D-amino acid oxidase) indirectly reduces NMDAR activity by degrading the NMDAR co-agonist D-serine. Since NMDAR hypofunction is thought to be a foundational defect in schizophrenia, hDAAO inhibitors have potential as treatments for schizophrenia and other nervous system disorders. Here, we sought to identify novel chemicals that inhibit hDAAO activity. We used computational tools to design a focused, purchasable library of compounds. After screening this library for hDAAO inhibition, we identified the structurally novel compound, ‘compound 2’ [3-(7-hydroxy-2-oxo-4-phenyl-2H-chromen-6-yl)propanoic acid], which displayed low nM hDAAO inhibitory potency (Ki=7 nM). Although the library was expected to enrich for compounds that were competitive for both D-serine and FAD, compound 2 actually was FAD uncompetitive, much like canonical hDAAO inhibitors such as benzoic acid. Compound 2 and an analog were independently co-crystalized with hDAAO. These compounds stabilized a novel conformation of hDAAO in which the active-site lid was in an open position. These results confirm previous hypotheses regarding active-site lid flexibility of mammalian D-amino acid oxidases and could assist in the design of the next generation of hDAAO inhibitors. Computational tools combined with experimental testing were used to identify novel inhibitors of hDAAO (human D-amino acid oxidase), an enzyme relevant for schizophrenia and neuropathic pain. These inhibitors stabilized a hDAAO conformation with the enzyme's active-site lid open.