Akshaya Murugesan , Aleksei Smirnov , Anxo Vila Alonso , Michela Buccioni , Chang Cui , Diego Dal Ben , Beatrice Francucci , Catia Lambertucci , Gabriella Marucci , Rosaria Volpini , Saravanan Konda Mani , Sandhanasamy Devanesan , Mohamad S. AlSalhi , Olli Yli-Harja , Andrea Spinaci , Meenakshisundaram Kandhavelu
{"title":"A2A receptor antagonist 4-(2-((6-Amino-9-ethyl-8-(furan-2-yl)-9H-purin-2-yl)amino)ethyl)phenol, a promising adenosine derivative for glioblastoma treatment","authors":"Akshaya Murugesan , Aleksei Smirnov , Anxo Vila Alonso , Michela Buccioni , Chang Cui , Diego Dal Ben , Beatrice Francucci , Catia Lambertucci , Gabriella Marucci , Rosaria Volpini , Saravanan Konda Mani , Sandhanasamy Devanesan , Mohamad S. AlSalhi , Olli Yli-Harja , Andrea Spinaci , Meenakshisundaram Kandhavelu","doi":"10.1016/j.ejps.2025.107039","DOIUrl":null,"url":null,"abstract":"<div><div>Adenosine, a pervasive signaling molecule mediated by its interaction with G-protein-coupled receptor subtypes, especially the A<sub>2A</sub> adenosine receptor (A<sub>2A</sub>AR), plays a crucial role in cancer treatment. Recently, A<sub>2A</sub>AR targeting adenosine analogs have been proposed as a potential therapeutic target for cancer treatment. However, the molecules targeting A<sub>2A</sub>AR and their mode of action in inhibiting glioblastoma cell progression remain unknown. We synthesized six adenosine derivatives substituted at the 9-, 2- and/or <em>N</em><sup>6-</sup> and/or 8- positions, and their anti-proliferative efficacy against the GBM cell lines LN229 and SNB19 was assessed. Molecular dynamic simulation integrated with experimental analyses, including cell cycle arrest, apoptosis assay, ligand binding assay, absorption, distribution, metabolism, excretion and toxicity (ADMET) profiling, PAMPA assay, and 3D spheroid analysis, were performed to identify the interaction efficacy of the potential derivative with A<sub>2A</sub>AR and its ability to prevent GBM cell progression. The most potent A<sub>2A</sub>AR derivative (ANR), 4-(2-((6-Amino-9-ethyl-8-(furan-2-yl)-9H-purin-2-yl)amino)ethyl)phenol (ANR 672) inhibits 5′-N-Ethylcarboxamidoadenosine (<strong>NECA</strong>)-induced cAMP validating the antagonistic property with higher cytotoxicity effect against GBM cells. ANR 672 showed higher affinity toward A<sub>2A</sub>AR (K<sub>i</sub> = 1.02 ± 0.06 nM) and exhibited significant IC<sub>50</sub> concentrations of ∼ 60–80 µM, than FDA approved drug istredefylline. The A<sub>2A</sub>AR-ANR 672 interaction profile showed well-defined hydrogen bonds and hydrophobic contacts, indicating a typical binding mechanism inside the receptor pocket and a higher degree of conformational flexibility than the A<sub>2A</sub>AR-Istradefylline complex. The antagonist effect of ANR 672 blocked the A<sub>2A</sub>AR signaling pathway, leading to necrosis-mediated cell death and cell cycle arrest at the S-phase in both the GBM cells. ANR 672 treated 3D tumour spheroids analysis with real-time spheroid volume and cell proliferation analysis revealed the potential ability of ANR 672 against GBM cell growth. Drug-likeness assessments also showed favorable pharmacokinetic profiles for ANR 672. Further validation of blood-brain barrier crossing potential revealed that ANR 672 possesses moderate permeability. Our findings shed light on how ANR 672 exerts its GBM-suppressive effect through the interaction of A<sub>2A</sub>AR. These preclinical results suggest that A<sub>2A</sub>AR blockade could be a unique strategy for treating GBM.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"207 ","pages":"Article 107039"},"PeriodicalIF":4.3000,"publicationDate":"2025-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"European Journal of Pharmaceutical Sciences","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0928098725000387","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"PHARMACOLOGY & PHARMACY","Score":null,"Total":0}
引用次数: 0
Abstract
Adenosine, a pervasive signaling molecule mediated by its interaction with G-protein-coupled receptor subtypes, especially the A2A adenosine receptor (A2AAR), plays a crucial role in cancer treatment. Recently, A2AAR targeting adenosine analogs have been proposed as a potential therapeutic target for cancer treatment. However, the molecules targeting A2AAR and their mode of action in inhibiting glioblastoma cell progression remain unknown. We synthesized six adenosine derivatives substituted at the 9-, 2- and/or N6- and/or 8- positions, and their anti-proliferative efficacy against the GBM cell lines LN229 and SNB19 was assessed. Molecular dynamic simulation integrated with experimental analyses, including cell cycle arrest, apoptosis assay, ligand binding assay, absorption, distribution, metabolism, excretion and toxicity (ADMET) profiling, PAMPA assay, and 3D spheroid analysis, were performed to identify the interaction efficacy of the potential derivative with A2AAR and its ability to prevent GBM cell progression. The most potent A2AAR derivative (ANR), 4-(2-((6-Amino-9-ethyl-8-(furan-2-yl)-9H-purin-2-yl)amino)ethyl)phenol (ANR 672) inhibits 5′-N-Ethylcarboxamidoadenosine (NECA)-induced cAMP validating the antagonistic property with higher cytotoxicity effect against GBM cells. ANR 672 showed higher affinity toward A2AAR (Ki = 1.02 ± 0.06 nM) and exhibited significant IC50 concentrations of ∼ 60–80 µM, than FDA approved drug istredefylline. The A2AAR-ANR 672 interaction profile showed well-defined hydrogen bonds and hydrophobic contacts, indicating a typical binding mechanism inside the receptor pocket and a higher degree of conformational flexibility than the A2AAR-Istradefylline complex. The antagonist effect of ANR 672 blocked the A2AAR signaling pathway, leading to necrosis-mediated cell death and cell cycle arrest at the S-phase in both the GBM cells. ANR 672 treated 3D tumour spheroids analysis with real-time spheroid volume and cell proliferation analysis revealed the potential ability of ANR 672 against GBM cell growth. Drug-likeness assessments also showed favorable pharmacokinetic profiles for ANR 672. Further validation of blood-brain barrier crossing potential revealed that ANR 672 possesses moderate permeability. Our findings shed light on how ANR 672 exerts its GBM-suppressive effect through the interaction of A2AAR. These preclinical results suggest that A2AAR blockade could be a unique strategy for treating GBM.
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