Insights into the Inhibitory Mechanisms of the Covalent Drugs for DNMT3A

The perturbations of DNA methyltransferase 3 alpha (DNMT3A) may cause uncontrolled gene expression, resulting in cancers and tumors. The DNMT inhibitors Azacytidine (AZA) and Zebu-larine (ZEB) inhibit the DNMT family with no specificities and consequently would bring side effects during the treatment. Therefore, it is vital to understand their inhibitory mechanisms in DNMT3A to inform the new inhibitor design for DNMTs. Herein, we carried out molecular dynamics (MD) and quantum mechanics/molecular mechanics (QM/MM) simulations to investigate the inhibitory mechanisms of AZA and ZEB. The results were compared to the methyl transfer of cytosine. We showed AZA might stop the methyl transfer process, whereas the ZEB might be stuck in a me-thyl-transferred intermediate (IM3). Such IM3 state then fails the elimination due to the unique protein dynamics that result in missing the catalytic water chain. Our results brought atomic-level insights into the mechanisms of the two drugs in DNMT3A, which could benefit the new generation of drug design for the DNMTs.