A general strategy to screen and evaluate antitussive, anti-inflammatory, and expectorant active components from raw...

Farfarae Flos and its honey-processed product are used to treat respiratory diseases; yet their active components and therapeutic targets remained unclear. Herein, we developed an integrated strategy combining receptor chromatography and mice models to screen and evaluate active components in both raw and honey-processed Farfarae Flos. We focused on three key receptors,β₂-AR, CysLT₁R, and M₃R, known to be involved in antitussive, anti-inflammatory, and expectorant activities. These receptors were immobilized onto silica gels to construct affinity columns. Using these columns, we identified the bioactive compounds in both forms of Farfarae Flos. The results revealed an identical set of target-specific compounds across raw and honey-processed samples: chlorogenic acid (CGA) and caffeic acid (CA) targeted β₂-AR; CGA and rutin bound to CysLT₁R; and CGA along with isochlorogenic acids A, B, and C interacted with M₃R. Notably, the interactions of rutin with CysLT₁R and isochlorogenic acids A, B, and C with M₃R are reported for the first time. These findings indicate that both raw and honey-fired Farfarae Flos share the same material basis for their antitussive, anti-inflammatory, and expectorant effects, with no significant alteration in pharmacological efficacy due to honey processing. Furthermore, the binding affinities of the bioactive compounds for the receptors correlated well with the molecular docking-predicted binding energies of the corresponding compound-receptor complexes. To our knowledge, this study represents the first target-centric comparative analysis of raw and processed Farfarae Flos. It establishes a generalizable framework for investigating processing-induced changes in herbal pharmacology and provides empirical evidence connecting metabolomic profiles to clinical efficacy.