Comprehensive phytochemical and pharmacological investigation of Cynoglossum clandestinum desf. ethanolic leaf extract: In vitro, In vivo, and In silico assessment of antioxidant, anti-inflammatory, and antidepressant properties.

ETHNOPHARMACOLOGICAL RELEVANCE: Cynoglossum clandestinum Desf. (C. clandestinum Desf.) is a wild Moroccan medicinal species traditionally used for wounds, burns, microbial infections, and inflammatory skin disorders.

AIM OF THE STUDY: This work aimed to characterize the phytochemical composition of the ethanolic leaf extract of C. clandestinum Desf. and to evaluate its antioxidant, anti-inflammatory, and antidepressant-like activities using in vitro, in vivo, and in silico approaches.

MATERIALS AND

METHODS

Leaves were macerated in absolute ethanol to obtain a reproducible ethanol-soluble fraction. Total phenolic and flavonoid contents were determined, and major compounds were identified by HPLC-DAD. Antioxidant activity was assessed by DPPH radical scavenging and total antioxidant capacity assays. Anti-inflammatory activity was evaluated in the carrageenan-induced paw edema model, and antidepressant-like activity was assessed in the forced swimming test in rats. Major identified compounds were further examined by ADMET prediction and molecular docking.

RESULTS

The extract contained 51.25 ± 1.06 mg GAE/g of total phenolics and 41.14 ± 1.81 mg QE/g of total flavonoids. HPLC-DAD identified six major phenolics, with quercetin-3-glucoside (12.08%) and epicatechin (10.17%) as the most abundant compounds. The extract showed strong antioxidant activity (DPPH IC50 = 0.290 ± 0.03 μg/mL; TAC = 295.83 ± 6.71 mg AAE/g), dose-dependent anti-inflammatory activity (63.64% edema inhibition at 400 mg/kg at T6), and a significant reduction in immobility time in the forced swimming test. Docking suggested quercetin-3-glucoside as the best xanthine oxidase binder, whereas quercetin showed the highest affinity for COX-2 and MAO-A.

CONCLUSION

The ethanolic leaf extract of C. clandestinum Desf. is rich in bioactive phenolics and shows promising antioxidant, anti-inflammatory, and antidepressant-like effects. Quercetin-3-glucoside appears to be the main antioxidant-oriented lead, whereas quercetin is the most promising anti-inflammatory and antidepressant-oriented lead. Further studies on isolated compounds, toxicity, and pharmacokinetics are required.