One destination, three routes: Tacrine-Celecoxib hybrids targeting cholinesterases, COX-2, and carbonic anhydrases in cognitive impairment linked to metabolic dysfunction.

In this study, a series of tacrine-celecoxib hybrid compounds was designed and synthesized to modulate key molecular targets implicated in metabolic dysfunction-associated mild cognitive impairment, including cyclooxygenase-2 (COX-2), carbonic anhydrases, and cholinesterases. Among the investigated derivatives, compound 10b was identified as the most balanced multitarget candidate, exhibiting selective inhibition of human butyrylcholinesterase (BuChE, IC50: 470 nM), potent COX-2 inhibitory activity (IC50: 50 nM), and low nanomolar Ki towards human carbonic anhydrase IX (hCA IX). Importantly, 10b showed lower neurotoxicity in neuronal cell models compared with tacrine and staurosporine. In a high-fat diet (HFD)-induced rat model of metabolic dysfunction and cognitive impairment, administration of 10b either orally or via intranasal nanoformulation significantly improved hippocampus-dependent cognitive performance as assessed by a composite behavioral z-score. Administration of 10b was also associated with improvements in glucose homeostasis, insulin sensitivity, lipid profile, together with a reduction of HFD-induced hepatological stress. Consistent with its multitarget profile, biochemical analyses showed significant attenuation of oxidative stress and neuroinflammatory markers in the hippocampus. Finally, docking and molecular dynamics simulation studies provided a structural rationale for the observed in vitro activities of 10b, highlighting favorable binding modes within the active sites of COX-2, BuChE, and hCA IX.