<i>In-Silico</i> Screening-Based Discovery of New Natural eEF2K Inhibitors with Neuritogenic Activity

Eukaryotic elongation factor 2 kinase (eEF2K), an atypical Ser/Thr-protein kinase that regulates neuronal protein synthesis homeostasis via an inhibitory phosphorylation of eEF2, has emerged as a promising therapeutic target for several diseases, including Alzheimer's disease (AD). In this study, we employed molecular docking with an in-house natural product library of 4270 compounds, containing 2177 novel compounds and 603 new structural frameworks, to identify eEF2K inhibitors. Following virtual screening, 25 natural products were selected for <i>in-vitro</i> evaluation of eEF2 phosphorylation inhibition as well as protein synthesis promotion. Our findings identified that compounds <b>17</b> and <b>23</b> potently suppress eEF2K activity, increase protein synthesis, and concurrently induce neuritogenesis. Molecular dynamics simulations suggest that <b>17</b> and <b>23</b> may stably bind to the eEF2K protein. Our findings highlighted <b>17</b> and <b>23</b> as new natural eEF2K inhibitors and promising candidates for promoting neural differentiation, providing potential therapeutic leads for the treatment of AD.