Herein, we report for the first time the G9a/EHMT2 inhibition and anti-Alzheimer's activities of the drug raltitrexed. G9a is a lysine methyltransferase that mainly dimethylates the H3K9 of chromatin, which triggers the repression of genes epigenetically, leading to various diseased conditions, including Alzheimer's disease (AD). First, we demonstrate that raltitrexed inhibits G9a at 120 nM. Moreover, raltitrexed lowers the total H3K9me2/H3K9 levels in AD transgenic C. elegans CL2006 worms, indicating that raltitrexed targets G9a directly. As toxicity is the bottleneck in G9a drug discovery, we conducted detailed in silico toxicity (TOPKAT) analyses of raltitrexed and measured the food consumption by C. elegans, demonstrating that raltitrexed's toxicity/function range is safe for the worm's growth. Moreover, we demonstrate that raltitrexed enhances the locomotive function of worms dose-dependently. Finally, we show that raltitrexed reduced the Aβ aggregates in worms up to 47%, highlighting the potential of raltitrexed in AD treatment.
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