Cömert Önder, F.Kahraman, N.Bellur Atıcı, E.Cagır, A.Kandemir, HakanTatar, G.Özpolat, Bülent2022-05-112022-05-1120212575-9108https://doi.org/10.1021/acsptsci.1c00030https://hdl.handle.net/20.500.11776/4705Eukaryotic elongation factor 2 kinase (eEF-2K) is an unusual alpha kinase involved in protein synthesis through phosphorylation of elongation factor 2 (EF2). eEF-2K is highly overexpressed in breast cancer, and its activity is associated with significantly shortened patient survival and proven to be a potential molecular target in breast cancer. The crystal structure of eEF-2K remains unknown, and there is no potent, safe, and effective inhibitor available for clinical applications. We designed and synthesized several generations of potential inhibitors. The effect of the inhibitors at the binding pocket of eEF-2K was analyzed after developing a 3D target model by using a domain of another ?-kinase called myosin heavy-chain kinase A (MHCKA) that closely resembles eEF-2K. In silico studies showed that compounds with a coumarin-chalcone core have high predicted binding affinities for eEF-2K. Using in vitro studies in highly aggressive and invasive (MDA-MB-436, MDA-MB-231, and BT20) and noninvazive (MCF-7) breast cancer cells, we identified a lead compound that was highly effective in inhibiting eEF-2K activity at submicromolar concentrations and at inhibiting cell proliferation by induction of apoptosis with no toxicity in normal breast epithelial cells. In vivo systemic administration of the lead compound encapsulated in single lipid-based liposomal nanoparticles twice a week significantly suppressed growth of MDA-MB-231 tumors in orthotopic breast cancer models in nude mice with no observed toxicity. In conclusion, our study provides a highly potent and in vivo effective novel small-molecule eEF-2K inhibitor that may be used as a molecularly targeted therapy breast cancer or other eEF-2K-dependent tumors. © 2021 American Chemical Society.en10.1021/acsptsci.1c00030info:eu-repo/semantics/openAccessapoptosisbreast cancercoumarinEF2Kelongation factor 2 kinasemolecular modelingchalconecoumarinelongation factor 2 kinaseimidazole derivativenanoparticlephosphotransferase inhibitorprotein kinase Bprotein p53animal cellanimal experimentanimal modelapoptosisArticleblood brain barrierbreast cancerBT-20 cell linecancer growthcancer survivalcardiotoxicitycell proliferationcolony formationcontrolled studycrystal structuredensitometrydrug designdrug synthesisenzyme active siteenzyme phosphorylationfemalein vitro studyin vivo studylipophilicityliposomal deliveryMCF-7 cell lineMDA-MB-231 cell lineMDA-MB-436 cell linemolecular dockingmolecular dynamicsmolecularly targeted therapymousenonhumanpharmacokinetic parameterspriority journalquantum mechanicstime-dependent inhibitiontumor xenograftWestern blottingArticle42926940N/AWOS:0006390672000472-s2.0-85105072435Q1