Bi Chunyang, Mirza Salahuddin, Baburi Helay, Schäkel Laura, Winzer Riekje, Moschütz Susanne, Keetz Kilian, Lopez Vittoria, Pelletier Julie, Sévigny Jean, Schulze Zur Wiesch Julian, Claff Tobias, Tolosa Eva, Namasivayam Vigneshwaran, Sträter Norbert, Müller Christa E
PharmaCenter Bonn, Pharmaceutical Institute, Department of Pharmaceutical & Medicinal Chemistry, University of Bonn, 53121 Bonn, Germany.
Department of Immunology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany.
ACS Pharmacol Transl Sci. 2025 Apr 3;8(5):1401-1415. doi: 10.1021/acsptsci.5c00126. eCollection 2025 May 9.
Ectonucleoside triphosphate diphosphohydrolase-1 (NTPDase1, CD39) catalyzes the extracellular hydrolysis of ATP generating AMP, while ecto-5'-nucleotidase (CD73) further hydrolyzes AMP yielding immunosuppressive adenosine. 8-Butylthioadenosine 5'-monophosphate (8-BuS-AMP) was described as a CD39 inhibitor but has been poorly characterized. The standard CD39 antagonist ARL 67156 is not suitable for in vivo studies due to metabolic instability. In the present study, we optimized and upscaled the synthesis of 8-BuS-AMP and performed a comprehensive investigation of its properties. It behaves as a competitive inhibitor at human and mouse CD39, and additionally inhibits CD73. Docking studies using a homology model of human CD39 and determination of an atomic-resolution (1.06 Å) cocrystal structure with human CD73 indicated the inhibitor's interactions within the substrate binding pockets and explained the compound's stability toward hydrolysis. 8-BuS-AMP is metabolically highly stable in human and mouse liver microsomes. It inhibited ε-adenosine formation from ε-ATP and ε-AMP in human synovial fluid and enhanced activation and proliferation of peripheral human T lymphocytes. Thus, 8-BuS-AMP is a recommended tool compound for studying purinergic signaling in vitro and in vivo, being superior to the standard CD39 inhibitor ARL 67156. Moreover, it may serve as a lead structure to develop drugs for the immunotherapy of cancer.
胞外核苷三磷酸二磷酸水解酶-1（NTPDase1，CD39）催化细胞外ATP水解生成AMP，而胞外5'-核苷酸酶（CD73）进一步将AMP水解产生具有免疫抑制作用的腺苷。8-丁硫基腺苷5'-单磷酸（8-BuS-AMP）被描述为一种CD39抑制剂，但对其特性的描述较少。标准的CD39拮抗剂ARL 67156由于代谢不稳定，不适合用于体内研究。在本研究中，我们优化并扩大了8-BuS-AMP的合成，并对其性质进行了全面研究。它在人和小鼠CD39上表现为竞争性抑制剂，此外还抑制CD73。使用人CD39的同源模型进行对接研究以及测定与人CD73的原子分辨率（1.06 Å）共晶体结构，表明了该抑制剂在底物结合口袋内的相互作用，并解释了该化合物对水解的稳定性。8-BuS-AMP在人和小鼠肝微粒体中代谢高度稳定。它抑制了人滑液中ε-ATP和ε-AMP生成ε-腺苷，并增强了人外周血T淋巴细胞的活化和增殖。因此，8-BuS-AMP是一种推荐的工具化合物，用于在体外和体内研究嘌呤能信号传导，优于标准的CD39抑制剂ARL 67156。此外，它可能作为一种先导结构来开发用于癌症免疫治疗的药物。
