Areceptor 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.

in European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences by 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

TLDR

  • The study synthesized and evaluated adenosine derivatives targeting the Aadenosine receptor (AAR) for glioblastoma (GBM) treatment, discovering a potent derivative (ANR 672) that inhibits AAR and exhibits anti-proliferative efficacy against GBM cells.

Abstract

Adenosine, a pervasive signaling molecule mediated by its interaction with G-protein-coupled receptor subtypes, especially the Aadenosine receptor (AAR), plays a crucial role in cancer treatment. Recently, AAR targeting adenosine analogs have been proposed as a potential therapeutic target for cancer treatment. However, the molecules targeting AAR and their mode of action in inhibiting glioblastoma cell progression remain unknown. We synthesized six adenosine derivatives substituted at the 9-, 2- and/or Nand/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 AAR and its ability to prevent GBM cell progression. The most potent AAR 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 AAR (K=1.02 ± 0.06 nM) and exhibited significant ICconcentrations of ∼ 60-80 µM, than FDA approved drug istredefylline. The AAR-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 AAR-Istradefylline complex. The antagonist effect of ANR 672 blocked the AAR 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 AAR. These preclinical results suggest that AAR blockade could be a unique strategy for treating GBM.

Overview

  • The study focuses on synthesizing adenosine derivatives targeting the Aadenosine receptor (AAR) and evaluating their anti-proliferative efficacy against glioblastoma (GBM) cell lines.
  • The study aims to identify the interaction efficacy of the derivatives with AAR and their ability to prevent GBM cell progression.
  • The primary objective is to explore ANR 672, the most potent AAR derivative, and its potential as a therapeutic agent for GBM treatment.

Comparative Analysis & Findings

  • The study synthesized six adenosine derivatives and evaluated their anti-proliferative efficacy against GBM cell lines LN229 and SNB19.
  • ANR 672, the most potent derivative, showed higher affinity toward AAR (K=1.02 ± 0.06 nM) and exhibited significant ICconcentrations of ∼ 60-80 µM.
  • The antagonist effect of ANR 672 blocked the AAR signaling pathway, leading to necrosis-mediated cell death and cell cycle arrest at the S-phase in both GBM cells.

Implications and Future Directions

  • The study's findings suggest that AAR blockade could be a unique strategy for treating GBM.
  • Future studies should focus on evaluating the efficacy and safety of ANR 672 in preclinical animal models and potentially human clinical trials.
  • Further research is needed to explore the pharmacokinetic profile of ANR 672 and its potential for blood-brain barrier crossing.
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