Subject description: Traditionally, it was thought that only human collagenases (matrix metalloproteinases-1, -8 and -13) were capable of initiating collagen degradation. Trypsin is also capable of cutting the triple helix of human collagens, and is at the root of rheumatoid arthritis.

Objectives: The trypsin inhibitor is considered a valid target for the discovery of new active compounds for the treatment of rheumatoid arthritis. In this study, a series of 59 compounds from the methanolic extract of A. leuchotruchus identified by GC-MS were used for a molecular docking study to identify interactions between compounds and active site amino acids.

Methods: Network pharmacology was adopted to detect the active components of our medicinal plants obtained from PubChem in 3D form, and the main target in the treatment of RA was obtained from the PDB. Key components and the target were selected for molecular anchoring.

Results and discussion: The evaluation of Ammodaucus leucotrichus seed extracts as potential anti-inflammatory and anti-arthritic agents involved employing methanol for the extraction of bioactive compounds. The extract underwent assessment through protease (trypsin) and protein (BSA, bovine serum albumin) denaturation inhibition assays. The methanol extract demonstrated trypsin inhibition of 85%, surpassing diclofenac (64.67%) at 125 μg/mL. GC–MS analysis revealed 59 and 58 secondary metabolites in the methanol extract, indicating diverse bioactive compounds. In silico docking studies identified 28 compounds with negative binding energies, suggesting potential trypsin inhibition. Notably, 2-hydroxyacetohydrazide showed superior inhibitory effects (−17.13 Kj/mol) compared to diclofenac (−13.01 Kj/mol). The methanol extract, particularly 2-hydroxyacetohydrazide, emerged as a promising candidate for rheumatoid arthritis (RA) treatment due to potent trypsin inhibition. SwissADME analysis highlighted favorable bioavailability attributes, with optimal size, polarity, solubility, and saturation for 2-hydroxyacetohydrazide. The BOILED-Egg model predicted blood–brain barrier permeation and gastrointestinal absorption, providing insights into druglikeness and bioavailability.

Conclusion: In summary, these findings propose that the methanol extract is a promising candidate for anti-inflammatory applications, particularly in trypsin inhibition. The identified compound, 2-hydroxyacetohydrazide, shows promise as a potential drug candidate for rheumatoid arthritis treatment. However, rigorous mechanistic studies and validation are crucial to enhance our understanding of its therapeutic potential within the field of plant-based medicines.