Benpyrine is a highly specific and orally active TNF-α inhibitor with a KD value of 82.1 μM and an IC50 value of 0.109 μM. It tightly binds to TNF-α, blocking its interaction with TNFR1, and has potential for TNF-α mediated inflammatory and autoimmune disease research [1].

TNF-α:82.1 μM (Kd)

Benpyrine (5-20 μM; 14 hours; RAW264.7 cells) pretreatment results in a dose-dependent decrease in the phosphorylation of IκBα in RAW264.7 cells (stimulated with 10 ng/mL TNF-α or 1 μg/mL LPS). Benpyrine abolishes the TNF-α-induced nuclear translocation of NF-κB/p65 in RAW264.7 cells[1].Benpyrine only blocks cell death induced by TNF-αWT and Y119A, and increases the cell survival rate up to 80%. Benpyrine does not obviously affect L57A- and Y59L-induced cytotoxicity in L929 cells[1].

Benpyrine (25-50 mg/kg; oral gavage; daily; for 2 weeks; Balb/c mice) significantly alleviates collagen-induced arthritis symptoms by dose-dependently decreasing proinflammatory cytokines (IFN-γ, IL-1β, IL-6) and increasing the anti-inflammatory cytokine IL-10. In a murine endotoxemia model, Benpyrine (25 mg/kg) reduces TNF-α-induced inflammation, thereby mitigating liver and lung injury[1].