Decoding the anti-inflammatory properties of sesquiterpene coumarins from the active fractions of Ferula assa-foetida through integrated experimental and in silico analyses

Ferula assa-foetida known as Asafoetida has a long-established history in folk medicine as a therapeutically useful drug in many disorders. The current paper explored the in vivo and in vitro anti-inflammatory activities of the plant. Liquid ethanol crude extract was subjected to liquid-to-liquid extraction, and the fractions so obtained were evaluated against the carrageenan-induced acute paw edema model in rats, to determine the anti-inflammatory effects. The fractions that gave the greatest activity-ethyl acetate and hexane- afforded four sesquiterpene coumarins: auraptene (1), umbelliprenin (2), galbanic acid (3), and kamolonol (4). The overall structure was determined through detailed spectroscopic investigation, ¹H NMR, ¹³C NMR, DPET135, COSY, HSQC, HMBC, HRESIMS, and other references to the literature. Evaluation of the in vitro activity of the LPS stimulated RAW 264.7 macrophages showed that the compound 4 (ethyl acetate fraction) exhibited maximum inhibitory action against the generation of nitric oxide (NO), followed by the compounds 1 and 2 of the hexane fraction. Network Pharmacology, Molecular Docking and Molecular dynamic simulation analysis were used to identify the target genes and explain the potential differences between the active compounds. As far as the authors know, there are no earlier reports that mention the anti-inflammatory action of kamolonol (4).