Network pharmacology and in vitro testing of Theobroma cacao extract’s antioxidative activity and its effects on cancer cell survival

Abstract

Theobroma cacao L. is a commercially important food/beverage and is used as traditional medicine worldwide against a variety of ailments. In the present study, computational biology approaches were implemented to elucidate the possible role of cocoa in cancer therapy. Bioactives of cocoa were retrieved from the PubChem database and queried for targets involved in cancer pathogenesis using BindingDB (similarity index ≥0.7). Later, the protein-protein interactions network was investigated using STRING and compound-protein via Cytoscape. In addition, intermolecular interactions were investigated via molecular docking. Also, the stability of the representative complex Hirsutrin-epidermal growth factor receptor (EGFR) complex was explored using molecular dynamics simulations. Crude extract metabolite profile was carried out by LC-MS. Further, anti-oxidant and cytotoxicity studies were performed in Chinese hamster ovary (normal) and Ehrlich ascites carcinoma (cancer) cell lines. Herein, the gene set enrichment and network analysis revealed 34 bioactives in cocoa targeting 50 proteins regulating 21 pathways involved in cancer and oxidative stress in humans. EGFR scored the highest edge count amongst 50 targets modulating 21 key pathways. Hence, it was selected as a promising anticancer target in this study. Structural refinement of EGFR was performed via all-atom molecular dynamics simulations in explicit solvent. A complex EGFR-Hirsutrin showed the least binding energy (-7.2 kcal/mol) and conserved non-bonded contacts with binding pocket residues. A stable complex formation of EGFR-Hirsutrin was observed during 100 ns MD simulation. In vitro studies corroborated antioxidant activity for cocoa extract and showed a significantly higher cytotoxic effect on cancer cells compared to normal cells. Our study virtually predicts anti-cancer activity for cocoa affected by hirsutrin inhibiting EGFR. Further wet-lab studies are needed to establish cocoa extract against cancer and oxidative stress.

Computational and experimental pharmacology to decode the efficacy of Theobroma cacao L. against doxorubicin-induced organ toxicity in EAC-mediated solid tumor-induced mice

“…In the present study, we observed the amelioration of various oxidative stress biomarkers in multiple organs after COE co-treatment with doxorubicin. Moreover, our previous studies also support the efficacy of COE in ameliorating oxidative stress and cancer progression (Patil et al, 2022b;2022a). Emmanuel et al reported that cocoa leaf polyphenolic-rich extract inhibited xanthine oxidase due to the flavonoids and phenolic acids present in the extract (Irondi et al, 2017).…”

“…We used the CastP tool to predict the largest and best possible binding pocket in both receptors, namely, LOX and XO. The parameters used for the virtual screening of phytochemicals against LOX and XO were selected from earlier similar studies ( Patil et al, 2022b ). The docked conformations obtained were ranked by the binding energy values, the intermolecular interactions were analyzed using Chimera, and publication-quality figures were generated in PyMOL.…”

“…About 27 previously reported phytocompounds in cacao were collated (Patil et al, 2022b). The successful docking of these 27 phytocompounds to both the receptors LOX and XO has been Frontiers in Pharmacology frontiersin.org performed using the POAP pipeline to the binding site determined by CastP.…”