Bononi Giulia, Gado Francesca, Masoni Samuele, Scalabrini Diana, Vedova Larissa Della, Di Stefano Miriana, Piazza Lisa, Poles Clarissa, Lonzi Chiara, Landucci Eva, Vagaggini Chiara, Poggialini Federica, Brai Annalaura, Caligiuri Isabella, Rizzolio Flavio, Mohamed Kawthar A, Bertini Simone, De Lorenzi Ersilia, Dreassi Elena, Macchia Marco, Minutolo Filippo, Laprairie Robert B, Granchi Carlotta, Tuccinardi Tiziano
Department of Pharmacy, University of Pisa, Via Bonanno 6, 56126 Pisa, Italy.
Department of Pharmaceutical Sciences, University of Milan, Via Mangiagalli 25, 20133 Milano, Italy; Department of Drug Sciences, University of Pavia, Via Taramelli 12, 27100 Pavia, Italy.
Bioorg Chem. 2025 Aug;163:108698. doi: 10.1016/j.bioorg.2025.108698. Epub 2025 Jun 19.
Monoacylglycerol lipase (MAGL) represents a key enzyme in the regulation of the endocannabinoid system and arachidonic acid signaling, emerging as a promising target for the treatment of inflammation, neurodegeneration, and cancer. Herein, we report the design, synthesis, and biological evaluation of a novel series of o-hydroxyanilide derivatives as reversible MAGL inhibitors. Starting from a known salicylketoxime scaffold, we introduced strategic modifications to reduce lipophilicity and enhance selectivity and potency. Among the synthesized compounds, 4-phenylbutanamide 40 exhibited the most potent inhibitory activity against human MAGL (IC = 0.34 μM), outperforming reference compound 8 (IC = 0.68 μM), and showing good selectivity over FAAH and cannabinoid receptors CB1R and CB2R. Biochemical characterization confirmed a reversible and competitive mechanism of inhibition. Molecular modeling studies supported the key interactions of compound 40 with the MAGL oxyanion hole and hydrophobic pocket. In cellular assays, compound 40 activated the Nrf2 antioxidant pathway and significantly inhibited NFκB-mediated inflammation, without inducing cytotoxic effects. ADME profiling revealed good membrane permeability and excellent plasma stability, despite moderate metabolic stability in human liver microsomes. Taken together, these findings highlight compound 40 as a promising reversible MAGL inhibitor endowed with anti-inflammatory and antioxidant properties, representing a valuable lead for the development of therapeutics targeting the endocannabinoid system.
单酰甘油脂肪酶（MAGL）是调节内源性大麻素系统和花生四烯酸信号传导的关键酶，已成为治疗炎症、神经退行性疾病和癌症的一个有前景的靶点。在此，我们报告了一系列新型邻羟基苯胺衍生物作为可逆MAGL抑制剂的设计、合成及生物学评价。从已知的水杨基酮肟支架出发，我们进行了策略性修饰以降低亲脂性并提高选择性和效力。在合成的化合物中，4-苯基丁酰胺40对人MAGL表现出最有效的抑制活性（IC = 0.34 μM），优于参考化合物8（IC = 0.68 μM），并且对脂肪酸酰胺水解酶（FAAH）以及大麻素受体CB1R和CB2R具有良好的选择性。生化特征证实了其可逆和竞争性的抑制机制。分子模拟研究支持了化合物40与MAGL氧阴离子空穴和疏水口袋的关键相互作用。在细胞试验中，化合物40激活了Nrf2抗氧化途径并显著抑制了NFκB介导的炎症，且未诱导细胞毒性作用。药物代谢动力学（ADME）分析显示尽管在人肝微粒体中的代谢稳定性中等，但具有良好的膜通透性和出色的血浆稳定性。综上所述，这些发现突出了化合物40作为一种有前景的可逆MAGL抑制剂，具有抗炎和抗氧化特性，是开发靶向内源性大麻素系统疗法的有价值先导化合物。
