Dilweg Majlen A, Gorostiola González Marina, de Ruiter Martijn D, Meijboom Nadine J, van Veldhoven Jacobus P D, Liu Rongfang, Jespers Willem, van Westen Gerard J P, Heitman Laura H, IJzerman Adriaan P, van der Es Daan
Division of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden University, PO Box 9502, Leiden, The Netherlands.
Oncode Institute, Leiden, The Netherlands.
Purinergic Signal. 2025 Apr;21(2):289-316. doi: 10.1007/s11302-024-10026-x. Epub 2024 Jun 15.
The human equilibrative nucleoside transporter 1 (SLC29A1, hENT1) is a solute carrier that modulates the passive transport of nucleosides and nucleobases, such as adenosine. This nucleoside regulates various physiological processes, such as vasodilation and -constriction, neurotransmission and immune defense. Marketed drugs such as dilazep and dipyridamole have proven useful in cardiovascular afflictions, but the application of hENT1 inhibitors can be beneficial in a number of other diseases. In this study, 39 derivatives of dilazep's close analogue ST7092 were designed, synthesized and subsequently assessed using [H]NBTI displacement assays and molecular docking. Different substitution patterns of the trimethoxy benzoates of ST7092 reduced interactions within the binding pocket, resulting in diminished hENT1 affinity. Conversely, [H]NBTI displacement by potentially covalent compounds 14b, 14c, and 14d resulted in high affinities (K values between 1.1 and 17.5 nM) for the transporter, primarily by the ability of accommodating the inhibitors in various ways in the binding pocket. However, any indication of covalent binding with amino acid residue C439 remained absent, conceivably as a result of decreased nucleophilic residue reactivity. In conclusion, this research introduces novel dilazep derivatives that are active as hENT1 inhibitors, along with the first high affinity dilazep derivatives equipped with an electrophilic warhead. These findings will aid the rational and structure-based development of novel hENT1 inhibitors and pharmacological tools to study hENT1's function, binding mechanisms, and its relevance in (patho)physiological conditions.
人类平衡核苷转运体1（SLC29A1，hENT1）是一种溶质载体，可调节核苷和核碱基（如腺苷）的被动转运。这种核苷调节多种生理过程，如血管舒张和收缩、神经传递和免疫防御。已证实诸如地拉卓和双嘧达莫等上市药物在心血管疾病中有用，但hENT1抑制剂的应用在许多其他疾病中可能有益。在本研究中，设计、合成了地拉卓的类似物ST7092的39种衍生物，随后使用[H]NBTI置换试验和分子对接进行评估。ST7092的三甲氧基苯甲酸酯的不同取代模式减少了结合口袋内的相互作用，导致hENT1亲和力降低。相反，潜在共价化合物14b、14c和14d引起的[H]NBTI置换导致对该转运体具有高亲和力（K值在1.1至17.5 nM之间），这主要是由于这些抑制剂能够以各种方式容纳在结合口袋中。然而，仍然没有与氨基酸残基C439共价结合的迹象，这可能是由于亲核残基反应性降低所致。总之，本研究引入了作为hENT1抑制剂具有活性的新型地拉卓衍生物，以及首个配备亲电弹头的高亲和力地拉卓衍生物。这些发现将有助于基于合理设计和结构的新型hENT1抑制剂以及用于研究hENT1功能、结合机制及其在（病理）生理条件下相关性的药理学工具的开发。
