Perillaldehyde-derived chromeno-dipyrimidines as multi-target-directed ligands: in vitro anticholinesterase and in vivo anti-inflammatory activities supported by molecular modeling.

In this work, eight novel chromeno-dipyrimidine derivatives were designed and synthesized through a green, catalyst-free one-pot hemisynthetic approach starting from the natural perillaldehyde, barbituric acid and substituted anilines. All prepared polycyclic heterocycles were isolated in high yields (74-94%), and were fully characterized by IR, 1H, 13C, 2D NMR spectroscopy and elemental analysis. Interestingly, the synthesized compounds underwent an unexpected intramolecular cyclization, leading exclusively to the formation of novel heterocyclic products, highlighting the structural versatility and reactivity of this system. The synthesized derivatives exhibited overall promising biological activity; notably, compounds BI-1, BI-2, and BI-3 emerged as the most active, displaying potent acetylcholinesterase inhibition (IC50 = 15.08-17.45 μM) together with moderate butyrylcholinesterase inhibition, while in vivo evaluation BI-3, BI-5, and BI-6 showed most significant anti-inflammatory effects (≤ 4.62%). Furthermore, docking studies performed on the chromeno-dipyrimidine derivatives suggest that the studied molecules exhibit inhibitory activity toward AchE and BchE, supported by their calculated docking scores. The findings from the docking analysis were further confirmed by 100 ns molecular dynamics simulations. The best-scoring complexes showed favorable dynamic behavior and stability when bound to both AChE and BChE during MD simulations.