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Fig 1.

Framework for DL-based virtual screening and predictive modeling of natural TNF-α inhibitors.

The model was developed using the ChEMBL dataset, with 2AZ5 as the target protein. After validation, the model was used to screen the Selleckchem Natural Compounds database. Compounds were prioritized based on their predicted pIC50 values, and after additional filtering, four compounds were chosen for MD simulation studies.

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Fig 1 Expand

Fig 2.

Complete dataset pre-processing pipeline.

Panel A represents the characterization of compounds using PubChem descriptors (881) of both training (ChEMBL) and testing (Selleckchem) datasets and Panel B depicts variance thresholding to retain essential features.

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Fig 2 Expand

Table 1.

Hyperparameter tuning of neural network architecture.

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Table 1 Expand

Fig 3.

Evaluation metrics of the DL regression model.

(a) MSE, (b) MAPE, (c) MAE, and (d) Loss.

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Fig 3 Expand

Fig 4.

Identification of screened compounds through Selleckchem’s natural product library along with their predicted pIC50 values.

The structures and the pIC50 values of the eligible candidates are included (CID 442977: Imperialine, CID 6070: Veratramine, CID 10098: Jervine, and CID 5390854: Gelsemine).

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Fig 4 Expand

Fig 5.

Visualization of docking scores and pIC50 values of 68 eligible natural candidates after drug-likeness analysis.

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Fig 5 Expand

Table 2.

ADME properties of the compounds Imperialine, Veratramine, Jervine, and Gelsemine using ADMETlab2.0 server.

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Table 2 Expand

Fig 6.

Docked poses of top hit compounds against target TNF-α.

(a) Imperialine, (b) Veratramine, (c) Jervine, and (d) Gelsemine.

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Fig 6 Expand

Table 3.

Drug-likeness analysis of final four compounds.

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Table 3 Expand

Fig 7.

2D molecular interaction of top hit compounds.

(a) Imperialine, (b) Veratramine, (c) Jervine, and (d) Gelsemine.

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Fig 7 Expand

Fig 8.

Retrospective validation of our virtual screening protocol.

a) ROC plot based on Tanimoto scores for MF and Layered fingerprint descriptors b) Heat map showing Tanimoto similarity between control (index 1), actives (index 2–21) and decoys (22–50) and c) Box plots with distribution of Tanimoto scores between active-control and decoy-control for both the fingerprint descriptors.

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Fig 8 Expand

Fig 9.

MD simulation analysis.

(a) RMSD, (b) RMSF, (c) Rg values plotted for the native apo-protein -black and apo-protein docked with selected bioactive molecules, (1) Imperialine-red, (2) Veratramine-green, (3) Jervine-yellow, and (4) Gelsemine- purple.

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Fig 9 Expand

Table 4.

Average structural and dynamic parameters for protein-ligand complex molecular dynamics simulation.

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Table 4 Expand

Table 5.

RMSF (nm) of the active side residues for the apoprotein and the docked complexes.

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Table 5 Expand

Fig 10.

MD simulation analysis.

(a) H-bonds, and (b) SASA values plotted for the native apo-protein (TNF-α in the human body)-black and apo-protein docked with selected bioactive molecules, (1) Imperialine-red, (2) Veratramine-green, (3) Jervine-yellow, and (4) Gelsemine–purple.

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Fig 10 Expand

Table 6.

The calculation of binding free energy results of 4 selected compounds.

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Table 6 Expand

Fig 11.

Principal component analysis of the MD simulation trajectories.

(a) 2AZ5, docked with (b) Imperialine, (c)Veratramine, (d) Jervine, and (e) Gelsemine.

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Fig 12.

Dynamic cross-correlation matrix of MD simulation trajectories.

2AZ5 complexed with (a) 2AZ5 apo form, (b) Imperialine, (c) Veratramine, (d) Jervine, and (e) Gelsemine on α-carbon atoms.

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Fig 12 Expand