Maspero Elena, Cappa Anna, Weber Janine, Trifirò Paolo, Amici Raffaella, Bruno Agostino, Fagà Giovanni, Cecatiello Valentina, Fattori Raimondo, Leuzzi Brian, Taibi Vincenzo, Meroni Giuseppe, Pasi Maurizio, Romussi Alessia, Sartori Luca, Villa Manuela, Vultaggio Stefania, Cirò Marco, Soffientini Paolo, Lombardo Lierin, Dahe Shakti, Bachi Angela, Varasi Mario, Rossi Mario, Pasqualato Sebastiano, Mercurio Ciro, Polo Simona
IFOM ETS, The AIRC Institute of Molecular Oncology, Milan, Italy.
Human Technopole, Milan, Italy.
Commun Chem. 2025 May 28;8(1):164. doi: 10.1038/s42004-025-01557-4.
Among the human E3 ubiquitin ligases, NEDD4 (Neural precursor cell expressed developmentally down-regulated 4) plays a critical role in development and cancer, making it a compelling therapeutic target. However, no specific NEDD4 inhibitors have advanced in drug development. In this study, we reveal the inhibitory mechanism of Norclomipramine, a tricyclic antidepressant, which inhibits NEDD4-mediated ubiquitin chain elongation by binding to a hydrophobic pocket in the Ub exosite of the N-lobe. Building on this mechanism, we conducted a focused medicinal chemistry campaign, resulting in the development of covalent inhibitors that specifically target the non-catalytic cysteine C627. These compounds exhibit selective binding to NEDD4 over other family members, effectively inhibiting NEDD4-mediated polyubiquitination while leaving monoubiquitinated substrates unaffected. Among these, compound 32 emerged as a potent lead (IC = 0.12 µM) with favorable pharmacokinetic properties, including oral bioavailability, paving the way for future in vivo efficacy studies.
在人类E3泛素连接酶中，NEDD4（神经前体细胞表达的发育性下调基因4）在发育和癌症中发挥着关键作用，使其成为一个极具吸引力的治疗靶点。然而，在药物开发中，尚未有特异性的NEDD4抑制剂取得进展。在本研究中，我们揭示了三环类抗抑郁药去甲氯米帕明的抑制机制，它通过与N端叶Ub外部位点的一个疏水口袋结合，抑制NEDD4介导的泛素链延长。基于这一机制，我们开展了一项有针对性的药物化学研究，开发出了特异性靶向非催化性半胱氨酸C627的共价抑制剂。这些化合物对NEDD4的结合选择性高于其他家族成员，能有效抑制NEDD4介导的多聚泛素化，而对单泛素化底物无影响。其中，化合物32成为了一种具有良好药代动力学性质（包括口服生物利用度）的强效先导物（IC = 0.12 μM），为未来的体内疗效研究铺平了道路。
