2.8.10. Determination of the Selected Phytoconstituents' Drug Abilities

The drug-likeness properties of the selected phytoconstituents were assessed using the Swiss ADME, pkCSM and PASS webservers [70–72]. The top 10 hit compounds' physicochemical or drug likeness properties demonstrated that most of the phytoconstituents followed Lipinski's rule. All the selected compounds' physiochemical parameters are shown in Table 5. The ADMET properties of the identified phytoconstituents were further investigated in order to rule out any potentially harmful patterns in the molecular structures (Table 6).

**Figure 7.** Comparative docking score analysis of the VDA proteins against top hit phytoconstituents.

**Figure 8.** Docking interaction analysis: 2D structural representation of Protein-ligand complexes having pi-Alkyl bonds (purple) and Hydrogen bond (green) along with interacted residues.


**Table 5.** The physicochemical properties of the selected top hit compounds.

#M.W. (Molecular Weight (Da); #Rot. bond (Rotatable bond); #HBA (hydrogen bond acceptor); #HBD (hydrogen bond donor).



GI abs. (Gastrointestinal absorption percentage); W.S. (Water Solubility (log mol/L); BBB/CNS permeation (blood brain barrier/central nervous system); Toxicity A/H/S (toxicity AMES/Hepatotoxicity/Skin sensitization).

A prediction of structure activity relations (SARs) was performed, by a machine learning program using the PASS online webserver, to investigate the biological activities of the selected phytoconstituents. *β*-amyrin was shown to have multiple biological activities, such as insulin promoter, caspase-3 stimulant, transcription factor NF kappa B stimulant, mucomembranous protector, hepatoprotection, apoptosis agonist, antineoplastic, oxidoreductase inhibitor, membrane integrity antagonist, and chemoprevention, with a Pa score ranging from 0.903 to 0.977. The biological activity predictions of all the ten phytoconstituents are provided in Table S8.

Moreover, *β*-amyrin, in combination with lupeol, was reported to have antibacterial activity, including antimycobacterial activity. The mixture of both compounds showed modest antibacterial activity against most of the bacteria, with MIC of 62.5 µg/mL, for *Staphylococcus aureus*, *Pseudomonas aeruginosa*, *Mycobacterium fortuitum* and *Mycobacterium smegmatis*. *β*-amyrin is abundantly found in plants with varied pharmacological activities. The compounds revealed after GC-MS analysis were used for molecular docking analysis with 10 shortlisted proteins. The docked complexes showed that most of the proteins had significant binding affinity with *β*-amyrin. Thus, *β*-amyrin was selected for further MD simulation study.
