N-acylethanolamines (NAEs), which include the endocannabinoid anandamide, represent an important family of signaling lipids in the brain. The lack of chemical probes that modulate NAE biosynthesis in living systems hamper the understanding of the biological role of these lipids. Using a high-throughput screen, chemical proteomics and targeted lipidomics, we report here the discovery and characterization of LEI-401 as a CNS-active N-acylphosphatidylethanolamine phospholipase D (NAPE-PLD) inhibitor. LEI-401 reduced NAE levels in neuroblastoma cells and in the brain of freely moving mice, but not in NAPE-PLD KO cells and mice, respectively. LEI-401 activated the hypothalamus-pituitary-adrenal axis and impaired fear extinction, thereby emulating the effect of a cannabinoid CB1 receptor antagonist, which could be reversed by a fatty acid amide hydrolase inhibitor. Our findings highlight the distinctive role of NAPE-PLD in NAE biosynthesis in the brain and suggest the presence of an endogenous NAE tone controlling emotional behavior.

中文翻译：

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发现可调节小鼠情绪行为的 NAPE-PLD 抑制剂。

N-酰基乙醇胺 (NAE)，包括内源性大麻素 anandamide，代表了大脑中一个重要的信号脂质家族。缺乏调节生命系统中 NAE 生物合成的化学探针阻碍了对这些脂质的生物学作用的理解。使用高通量筛选、化学蛋白质组学和靶向脂质组学，我们在此报告 LEI-401 作为 CNS 活性 N-酰基磷脂酰乙醇胺磷脂酶 D (NAPE-PLD) 抑制剂的发现和表征。LEI-401 降低了神经母细胞瘤细胞和自由移动小鼠大脑中的 NAE 水平，但分别降低了 NAPE-PLD KO 细胞和小鼠的 NAE 水平。LEI-401 激活下丘脑-垂体-肾上腺轴并削弱恐惧消退，从而模拟大麻素 CB1 受体拮抗剂的作用，这可以被脂肪酸酰胺水解酶抑制剂逆转。我们的研究结果强调了 NAPE-PLD 在大脑中 NAE 生物合成中的独特作用，并表明存在控制情绪行为的内源性 NAE 基调。