SPR‐UPLC‐QTOF/MS‐GNPS‐Guided Discovery of Hypermonol J as a Novel Tumor Necrosis Factor‐Binding Small‐Molecule Lead for Inflammatory Storm Intervention

ABSTRACT This study developed an integrated strategy combining surface plasmon resonance (SPR) for real‐time biomolecular interaction analysis, ultra‐performance liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry (UPLC‐QTOF/MS) for compound profiling, and Global Natural Products Social Molecular Networking (GNPS)‐based molecular networking for structural annotation in complex herbal systems. Using this platform, we identified hypermonol J (HPJ), a polycyclic polyprenylated acylphloroglucinol (PPAP), as a high‐affinity small‐molecule binder of tumor necrosis factor (TNF) ( K D = 25.9 pM). SPR and orthogonal isothermal titration calorimetry (ITC) supported direct TNF binding, whereas mechanistic assays suggested that HPJ may preferentially stabilize TNF dimers, thereby impairing trimer assembly and downstream TNF receptor 1 (TNFR1)/nuclear factor kappa B (NF‐κB) signaling. In a preclinical psoriasis model, topical 1% HPJ alleviated imiquimod (IMQ)‐induced epidermal hyperplasia and suppressed IL‐17/IL‐23 pathways. In an acute lung injury (ALI) model, 10 mg/kg HPJ reduced neutrophil infiltration, lowered serum inflammatory cytokine levels, and restored pulmonary tight junction integrity. Biosafety assessments supported the preliminary tolerability of HPJ under the tested conditions. These findings identify HPJ as a promising small‐molecule lead for further pharmacological optimization and underscore the value of SPR‐guided natural product discovery for inflammatory storm intervention.