Network pharmacology and virtual screening suggest neoprzewaquinone a as a potential therapeutic agent for rheumatoid arthritis, validated in fibroblast-like synoviocytes and collagen-induced arthritis models.

Currently, utilized disease-modifying antirheumatic drugs (DMARDs) do not provide a complete cure for rheumatoid arthritis (RA) and are associated with side effects related to prolonged use. In contrast, natural products have demonstrated promising efficacy and low toxicity in treating various diseases. This study employed network pharmacology and virtual screening to identify natural products with potential therapeutic effects for RA, followed by molecular dynamics simulations and experimental validation. Through network pharmacology, we identified interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) as RA-related targets, and subsequently prioritized neoprzewaquinone A (NEO) via virtual screening and molecular dynamics simulations for further validation. Subsequent experiments indicated that NEO could inhibit the binding of IL-1β, IL-6, and TNF-α to their respective receptors, reduce proliferation, and downregulate the migratory and invasive capacities of RA fibroblast-like synoviocytes (RA-FLS). Furthermore, NEO alleviated RA-related symptoms and diminished tissue damage in collagen-induced arthritis (CIA) mice. RNA sequencing suggested that the alleviative effect of NEO on RA may be attributed to the inhibition of the activation of the MAPK signaling pathway. Additionally, NEO appeared to reduce the phosphorylation levels of MEK and ERK. Importantly, safety evaluations did not reveal a clear increase in liver or kidney toxicity associated with NEO, within the scope of this study. Collectively, these findings suggested that NEO shows promise as a potential natural therapeutic option for RA and offers a new direction and theoretical foundation for future pharmacotherapy research in this area.