Molecular mechanisms of phytoconstituents from selected Egyptian plants against non-small cell lung cancer using integrated in vitro network pharmacology and molecular docking approach
Authors: Shaala Lamiaa A. , Youssef Diaa T. A. , Ramadan Mahmoud A. , Khalifa Azza A. , Ibrahim Reham S. , Valeriote Fred , Celik Ismail , Dawood Hend M. 
Published: January 2025

Abstract
Non-small cell lung cancer (NSCLC) is a widespread highly malignant type of lung cancer. Conventional chemotherapeutic drugs may be accompanied by both drug resistance and serious side effects in patients. Therefore, safer and more effective medications are urgently needed for the treatment of NSCLC. This study investigates the mode of action of 21 phytoconstituents previously isolated from the Amaryllidaceous plants Crinum bulbispermum (Burm.f.), Pancratium maritimum L., and Hippeastrum vittatum Herbert alongside the Asteraceous plant Centaurea scoparia Sieb. for therapy of NSCLC via in vitro cytotoxic, network pharmacology, and molecular docking analyses. Despite the in vitro and in vivo cytotoxic studies carried out on phytoconstituents from these plants in treating numerous cancer types, scarce information documenting their cytotoxic activity towards NSCLC cells is available. First, the compounds were tested for their in vitro cytotoxic activities and selectivity on human non-small cell lung cancer cells using disk diffusion assay. Compounds having significant potencies were promoted for network pharmacology analysis. Pharm mapper, Genecards, STRING, and KEGG databases were utilized for surfing target genes and pathways for these compounds, while for construction of compound-target-pathway (C-T-P) network, Cytoscape 3.7.1. freeware was used. Molecular docking and dynamics simulation were run for the top hit constituents against the most enriched molecular targets followed by in silico ADMET studies using Schrodinger® suite and Gromacs. In vitro cytotoxicity testing demonstrated that crinamine was the most potent compound followed by lycorine, hemanthidine, and haemanthamine. The network pharmacology approach revealed the enrichment of acetyllycoramine, pluviine, 5-hydroxy-7-methoxy-2-methylchromone, and ismine. Whereas, androgen receptor (AR), epidermal growth factor receptor (EGFR), and estrogen-sensitive receptor alpha (ESR1) were the most enriched target genes. Pathway analysis revealed that central carbon metabolism, EGFR tyrosine kinase inhibitor endocrine resistance, and non-small cell lung cancer were the most enriched cancer-related pathways. Ismine possessed the most stable ligand-protein interactions when docked to the three proteins, with MD simulations further confirming its strong and consistent binding to AR, moderate stability with ESR-1, and lower stability with EGFR over the 100 ns trajectory. ADMET study conducted on the above compounds confirmed their excellent drug-likeness properties, oral bioavailability, and safety profiles highlighting the need for some structural modifications to pluviine to enhance its oral bioavailability. These integrated approaches showed that some constituents from the investigated plants interact synergistically against non-small cell lung cancer-related genes and pathways.