The cytotoxic effects of the extract were investigated on 3 human cancer cell lines i.e. the A549 non-small-cell lung carcinoma (NSCLC), the MCF7 breast adenocarcinoma and the U373 glioblastoma using a MTT colorimetric assay. Biological data allowed to guide the fractionation of the extract by separation and purification on silica gel 60 (CC and TLC). The isolated compounds which were characterized by spectral analysis, mainly HR-ESIMS, HR-EIMS, UV and NMR experiments (1H, 13C, COSY, ROESY, HSQC and HMBC) and comparison of their spectroscopic data with those reported in the literature, were evaluated for cytotoxic activities on six cancer cell lines (A549, MCF7, U373, Hs683 human glioma, PC3 human prostate and B16-F10 murine melanoma). The direct and indirect antibacterial and antifungal activities were determined using microdilution methods for the raw extract and TLC-bioautography and microdilution methods against standard and clinical strains for the isolated compounds.
The raw extract reduced cell viability with IC50s of 27, 25 and 21 μg/mL on A549, MCF7 and U373, respectively. Five secondary metabolites: two phenolic compounds (vanillin lass="boldFont">1, paridol lass="boldFont">3), a lignan [(−)-arctigenin lass="boldFont">2] and two flavonoid aglycones (eupatilin lass="boldFont">4 and jaceosidin lass="boldFont">5), were then isolated from this extract. Moderate cytotoxic effects were observed for (−)-arctigenin lass="boldFont">2 (IC50s: 28 and 33 μM on Hs683 and B16-F10, respectively), eupatilin lass="boldFont">4 (IC50s: 33 and 47 μM on B16-F10 and PC3, respectively) and jaceosidin lass="boldFont">5 (IC50s: 32 and 40 μM on PC3 and B16-F10, respectively).
All the isolated compounds were described for the first time from this species. Although inactive against 7 tested microorganisms (fungi, bacteria and yeast, human or plant pathogens), the raw extract was able to potentiate the effect of beta-lactam antibiotics on methicillin-resistant Staphylococcus aureus (MRSA), reducing the minimal inhibitory concentrations (MICs) by a factor of 2-32-fold. No synergy was found between the extract and streptomycin. From the five isolated compounds only jaseosidin lass="boldFont">5 showed a moderate antimicrobial activity.