Anti-inflammatory potential of isoquinoline alkaloids from Fumaria officinalis: in vitro, in vivo, and in silico evaluation

The objective of this study is to identify the alkaloids of Fumaria officinalis and to evaluate their anti-inflammatory activity through three approaches in vitro, in vivo and in silico. In vitro and in vivo anti-inflammatory activities were evaluated using the BSA denaturation method and induction of paw edema by carrageenan. In silico molecular docking and ADME/T studies were carried out to evaluate the potential of the identified alkaloid compounds against cyclooxygenase-II (COX-2) enzyme. The LC-MS/MS analysis results revealed the presence of 17 isoquinoline alkaloids, among which jatrorrhizine and protopine were the most abundant, with percentages of 29.29 and 8%, respectively. The extract of F. officinalis demonstrated maximum BSA protection at a concentration of 500 µg/mL (76.16% efficiency). Furthermore, the extract showed a dose-dependent anti-inflammatory effect on carrageenan-induced paw edema, with the best effect observed at the 6th hour in mice treated with 200 mg/kg of alkaloid extract (77% efficiency). Molecular docking revealed that protopine, bicuculline, stylopine, and coptisine exhibited high affinity to the receptor (with high interaction energy by around 10), with strong hydrogen and hydrophobic bonds. Stylopine fulfilled all ADME/T conditions and did not present any toxicity. In addition, quantum chemical analysis of stylopine confirmed its electronic stability and reactivity, supporting its potential as a COX-2 inhibitor. These findings demonstrate that the stylopine could be considered a powerful anti-inflammatory compound.