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Fig 1.

Chemical structures of Donepezil, Galantamine and five hit natural molecules investigated in this work as potential AChE inhibitors.

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Fig 1 Expand

Table 1.

Molecular properties of hit natural molecules compared to FDA approved AChE inhibitors.

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Table 1 Expand

Fig 2.

Docked conformations and important binding site interactions of suggested natural compounds as compared to control molecules.

Panels A-B for Donepezil (3D-2D), C-D for Galantamine (3D-2D), E-F for Queuine (3D-2D) and G-H for Etoperidone (3D-2D) respectively. Coloring indicates: CAS (magenta), PAS site (cyan), anionic site (purple), and acyl binding site (orange).

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Fig 2 Expand

Table 2.

Docking analysis of proposed natural compounds as compared with control molecules: Docking scores, interaction sites and residues involved in AChE binding site.

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Table 2 Expand

Fig 3.

Molecular dynamics results of Queuine: RMSD (A), RMSF (B), protein-ligand fractions (C), protein-ligand contacts (% of simulation time) (D), SASA, radius of gyration and intramolecular hydrogen bonding (E) plots for Queuine.

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Fig 3 Expand

Fig 4.

Molecular dynamics results of Etoperidone: RMSD (A), RMSF (B), protein-ligand fractions (C), protein-ligand contacts (% of simulation time) (D), SASA, radius of gyration and intramolecular hydrogen bonding (E) plots for Etoperidone.

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Fig 4 Expand

Fig 5.

Final snapshots (100 ns) obtained from MD simulations: Etoperidone (A) and Queuine (B).

Etoperidone stability is maintained within the binding cleft and interacts with CAS site (gray meshed surface); Queuine seems to diffuse out of the binding cleft and preserves its interactions with the PAS site.

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Fig 5 Expand

Table 3.

MM-GBSA calculations from MD simulations.

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Table 3 Expand

Table 4.

Predicted druglike and ADMET properties of hit natural AChE inhibitor molecules compared to donepezil and galantamine.

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Table 4 Expand

Fig 6.

The results of the RTCA analysis: The cells were not treated with any drug for the first 24 h.

After 24 hour, different concentrations of donepezil (A), galantamine (B), queuine (C), etoperidone (D), and thiamine (E) were applied. Following drug application, cell index was measured over time for 48 hours and plotted.

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Fig 6 Expand

Fig 7.

Percentage graph of cell viability of SH-SY5Y cells compared to the control group after exposure to different concentrations: Donepezil (A), Galantamine (B), Queuine (C), Etoperidone (D), and Thiamine (E) for 48 hours.

The results represent the average ± SEM of the results from each experiment, which were repeated three times at different times with the same concentration ranges. Compared to the control, *P\0.05, **P\0.01, ***P\0.001.

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Fig 7 Expand

Table 5.

IC50 values for selected test compounds as AChE inhibitors.

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Table 5 Expand

Fig 8.

Schematic representation of the workflow.

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