Murugesan Akshaya, Alonso Anxo Vila, Konda Mani Saravanan, Sarkar Puja, Smirnov Aleksei, Francucci Beatrice, Marucci Gabriella, Buccioni Michela, Abass Kasim S, Sureka Chandrabose, Yli-Harja Olli, Kandhavelu Meenakshisundaram
Molecular Signaling Group, Faculty of Medicine and Health Technology, Tampere University, Tampere, 33710, Finland; BioMeditech and Tays Cancer Center, Tampere University, Hospital, P.O. Box 553, 33101, Tampere, Finland.
B-Aatral Biosciences Private Limited, Bangalore, 560091, Karnataka, India.
Eur J Pharmacol. 2025 Sep 5;1002:177864. doi: 10.1016/j.ejphar.2025.177864. Epub 2025 Jun 20.
Glioblastoma multiforme (GBM), a multifactorial deadliest cancer with constrained clinical efficacy due to heterogeneity, drug resistance, side effects of the chemotherapeutic drug, necessitating the development of novel cancer therapeutics. The adenosine A receptor targeted binding of antagonist leads to regulation of downstream effectors, mediating the phosphorylation of Casein kinase 1δ kinase domain (CK1δ) in cancers. Here, we performed a comparative investigation of Food and Drug Administration (FDA) approved drugs, istradefylline and riluzole inhibiting adenosine A receptor and CK1δ isoform in GBM cell growth. Molecular interaction of riluzole with CK1δ isoform and istradefylline with adenosine A receptor was identified through molecular docking and dynamic simulations. The potential of these two FDA approved drugs in inhibiting GBM cell growth was investigated through various in-vitro analysis including dose-dependent dynamic assay, cell cycle assay, apoptosis assay by flow cytometry. Further the effect of these drugs on spheroid cell growth and cell size was measured. In silico analyses demonstrated that riluzole binds strongly to CK1δ isoform with a binding energy of -9.02 kcal/mol, whereas istradefylline binds to adenosine A receptor with -9.88 kcal/mol. In vitro evaluation revealed that riluzole increased cell growth inhibition by 24 % in LN229 cells and 36 % in SNB19 cells than istradefylline. Riluzole arrested the cell cycle at S phase in both cell lines, whereas istradefylline arrest was cell line specific. Three-dimensional (3D) spheroid model of 1321N1 GBM cells further demonstrated that riluzole inhibits ∼50 % higher cell growth inhibition than istradefylline with effective reduction in spheroid volume and size. Overall, our analysis revealed that blocking of adenosine A receptor downstream signaling pathway protein CK1δ with its inhibitor, riluzole, showed higher anti-GBM effect than its upstream signaling blocker, istradefylline. Thus, blocking adenosine A receptor downstream effector signaling protein through its antagonist and blocking its effector protein CK1δ could provide an opportunity to develop targeted therapy for GBM.
多形性胶质母细胞瘤（GBM）是一种多因素导致的最致命癌症，由于其异质性、耐药性以及化疗药物的副作用，临床疗效有限，因此需要开发新型癌症治疗方法。腺苷A受体拮抗剂的靶向结合可导致下游效应器的调节，介导癌症中酪蛋白激酶1δ激酶结构域（CK1δ）的磷酸化。在此，我们对美国食品药品监督管理局（FDA）批准的药物——异他林和利鲁唑抑制腺苷A受体和CK1δ异构体在GBM细胞生长中的作用进行了比较研究。通过分子对接和动态模拟确定了利鲁唑与CK1δ异构体以及异他林与腺苷A受体的分子相互作用。通过各种体外分析，包括剂量依赖性动态测定、细胞周期测定、流式细胞术凋亡测定，研究了这两种FDA批准的药物抑制GBM细胞生长的潜力。此外，还测量了这些药物对球体细胞生长和细胞大小的影响。计算机分析表明，利鲁唑以-9.02千卡/摩尔的结合能与CK1δ异构体强烈结合，而异他林以-9.88千卡/摩尔的结合能与腺苷A受体结合。体外评估显示，与异他林相比，利鲁唑使LN229细胞的细胞生长抑制增加了24%，使SNB19细胞的细胞生长抑制增加了36%。利鲁唑使两种细胞系的细胞周期停滞在S期，而异他林的停滞具有细胞系特异性。1321N1 GBM细胞的三维（3D）球体模型进一步表明，利鲁唑比异他林抑制细胞生长的效果高约50%，并有效降低了球体体积和大小。总体而言，我们的分析表明，用其抑制剂利鲁唑阻断腺苷A受体下游信号通路蛋白CK1δ，比其上游信号阻断剂异他林显示出更高的抗GBM效果。因此，通过其拮抗剂阻断腺苷A受体下游效应器信号蛋白并阻断其效应蛋白CK1δ，可能为开发GBM的靶向治疗提供机会。
