In silico virtual screening of natural small molecules for dual inhibition of Aβ and tau aggregation in Alzheimer's disease

Alzheimer's disease (AD) is defined by the accumulation of amyloid beta (Aβ) plaques and twisted fibers of hyperphosphorylated tau, with their aggregation driving neurodegeneration and cognitive decline. Presently, the majority of therapies in development are largely single-targeted, focusing either on Aβ or tau aggregation. This highlights a significant gap in strategies that can simultaneously modulate aggregation of both proteins in AD. This study employed an integrated in silico approach to identify phenolic small molecules capable of modulating both Aβ and tau aggregation. From a library of 160 natural compounds, fifteen phenolic-structured compounds inhibited Aβ aggregation by more than 50 %, and subsequent in silico screening identified baicalein, genkwanin, 2-hydroxycinnamaldehyde, and gallic acid as potential dual inhibitors. To further evaluate their dual-modulating potential, we conducted molecular docking, molecular dynamics simulations, and pharmacokinetic predictions. First, molecular docking with multiple conformers of Aβ and tau showed that all four compounds bound strongly to key aggregation-prone residues. Second, molecular dynamics simulations further demonstrated stable interactions with dimeric Aβ and tau. Third, pharmacokinetic predictions supported drug-likeness, including favorable blood-brain barrier permeability and systemic clearance. Finally, Thioflavin T assays were subsequently conducted to examine whether the computationally predicted anti-aggregation potential was reflected in biological outcomes, and the results confirmed that the predicted interactions were partially recapitulated under in vitro conditions. Together, our findings provide mechanistic and pharmacokinetic support for the further development of these phenolic compounds as Aβ and tau-targeting agents in early-stage AD therapy.