Design, synthesis, and biological evaluation of pyranone-carbamate hybrids as selective butyrylcholinesterase inhibitors with anti-neuroinflammatory activity for Alzheimer's disease.

Alzheimer's disease (AD) is a multifactorial neurodegenerative disorder in which cholinergic dysfunction and chronic neuroinflammation jointly contribute to cognitive decline. To address these converging pathological features, a series of carbamate-pyranone hybrids (E1-E17) were designed using a pharmacophore hybridization strategy that integrates the cholinesterase-inhibitory carbamate motif of Rivastigmine with the anti-neuroinflammatory γ-pyranone scaffold derived from D30. Among these derivatives, E14 emerged as a promising hit compound. E14 inhibited lipopolysaccharide-induced nitric oxide production in BV2 microglial cells (IC₅₀ = 9.76 ± 0.59 μM), while maintaining acceptable cytocompatibility. Enzymatic assays revealed that E14 selectively inhibited butyrylcholinesterase, with IC₅₀ values of 15.59 ± 2.98 μM against eqBuChE and 38.65 ± 3.40 μM against hBuChE, while showing negligible activity against acetylcholinesterase, and kinetic analysis indicated a competitive inhibition mechanism. Consistent with its CNS-oriented design, E14 exhibited high passive blood-brain barrier permeability in a PAMPA-BBB assay (Pe = 11.87 × 10-6 cm/s) and showed good acute tolerability in mice. In an oligomeric Aβ-induced cognitive impairment mouse model, E14 significantly improved recognition memory and spatial learning performance. Mechanistic investigations demonstrated that E14 attenuated hippocampal glial activation and neuroinflammation, suppressing the TLR4/p38 MAPK signaling pathway and modulating the IL-1β/C3-mediated microglia-astrocyte inflammatory axis. Network pharmacology analyses further suggested multitarget engagement across inflammation- and stress-related pathways relevant to AD pathology. Collectively, these findings identify E14 as a brain-penetrant, BuChE-selective dual-functional compound that integrates cholinesterase inhibition with anti-neuroinflammatory activity, supporting its further development as a multitarget-directed lead for AD intervention.