Evaluation of the phytochemical profile, biological activities, and acute toxicity of Cistus ladanifer aerial part extracts: experimental and in silico investigations

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Introduction Using both experimental and in silico approaches, this study aimed to evaluate the phytochemical composition and biological activities of Cistus ladanifer aerial part extracts. Methods Cistus ladanifer aerial parts were collected in the Khenifra region, Morocco. Hydroethanolic (Soxhlet) and aqueous (decoction) extracts were prepared. Physicochemical parameters, mineral content (ICP-AES), phytochemical screening, total polyphenols, flavonoids, and condensed tannins were determined. HPLC/UV-ESI-MS was used for chemical profiling. Antioxidant activity was assessed by TAC, DPPH, and FRAP assays; antimicrobial activity by MIC and MBC/MFC against bacterial and fungal strains; acute toxicity in mice at 2 g/kg; antihyperglycemic activity by oral glucose tolerance test in rats; and molecular docking against selected antioxidant, antimicrobial, and antihyperglycemic protein targets. Results Physicochemical analysis of the plant material showed a moisture content of 12.68% ± 0.42%, pH of 4.16 ± 0.05, and ash content of 5.50% ± 0.31%. Mineral analysis revealed essential elements such as Zn, Mn, Fe, and Al. Phytochemical screening confirmed major secondary metabolites, including flavonoids, tannins, saponins, and alkaloids. The hydroethanolic extract exhibited the highest extraction yield (23.81% ± 0.63%), whereas the aqueous extract ( CL2 ) was richer in total polyphenols (433.89 ± 1.82 mg GAE/g DE), flavonoids (54.13 ± 0.52 mg CE/g DE), and condensed tannins (19.50 ± 0.87 mg CE/g DE). HPLC/UV-ESI-MS analysis revealed the presence of several phenolic compounds, including resveratrol 4′-(6-galloylglucoside), kaempferol, and quercetin 3.4′-diglucoside. CL2 exhibited strong antioxidant activity with a total antioxidant capacity of 462.04 ± 0.41 mg EAA/g DE, a DPPH IC50 of 7.06 ± 0.31 μg/mL, and a FRAP EC50 of 10.42 ± 0.83 μg/mL. Antimicrobial assays showed notable inhibitory effects against several microorganisms, particularly Candida kefyr and Saccharomyces cerevisiae (MIC = 4.69 mg/mL). In addition, CL2 demonstrated significant antihyperglycemic activity in vivo and showed no observable signs of toxicity or mortality at an oral dose of 2 g/kg. Molecular docking analysis further reinforced these results by showing favorable interactions of major phytochemicals with target proteins associated with the investigated biological activities. Discussion Our findings highlight C. ladanifer as a potent source of bioactive compounds with promising therapeutic applications.

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