Abstract
Fungi, or mushrooms, are a source of bioactive compounds that have valuable health effects. The column chromatography of ethanolic extract of Daldinia eschscholtzii (DEE) was performed, 10 fractions were collected, and out of them the F5 fraction demonstrated the greatest antioxidant activity by DPPH (1,1 diphenyl-2-picrylhydrazyl) with an EC50 of 0.94 ± 0.05a mg/mL. Mycochemistry analysis by liquid chromatography mass spectrum (LC-MS) showed that F5 was a mixture of four compounds, like myricetin, tricin, eudesmin, and (-)-curcumol. Cytotoxicity assay exhibited anti-proliferation activity with IC50 values of 125.66 ± 6.34, 96.22 ± 1.92, and 76.91 ± 2.05 µg/mL of F5 against A549 lung cancer cell line at 24, 48, and 72 h, respectively. The mechanism of anticancer effect of F5 (DEE) on the cancer cell line (A549) included the induction of apoptosis, reduction of mitochondrial potentiality, and change of gene expression levels of TCASPASE 3, TCASPASE 9, TP53, and TBCL2 of the cell line. The qPCR study showed that TCASPASE 3, 9, TBAX, and TP53 were up-regulated significantly, whereas TBCL2 was down-regulated significantly after the treatment with F5 (50 and 150 µg/mL) for 24 h. It had an anti-migration activity, which was justified by downregulation of TMMP 2 & 9, and TVEGF genes. The physicochemical, pharmacokinetic, and medicinal properties of all four compounds in F5 were screened by three web predictors (SwissADME, pkCSM, and AdmetSAR), and among them, curcumol, tricin, and eudesmin compounds fulfilled all the criteria of different rules or filters (Lipinski's rules, Ghose filter, Veber filter, Egan filter, and Muegge filter) for drug development. Molecular docking studies of the compounds like tricin, curcumol, and eudesmin with receptor proteins like Caspase 3, p53, and BCL2 revealed that tricin had the highest binding energy towards Caspase 3 and p53, allowing upregulation of these genes, while eudesmin showed the highest binding energy (-7.6 kcal/mol) with BCL2, suggesting inactivation of BCL2. In conclusion, F5 of DEE had both anticancer and anti-metastasis properties.