Phytomedicine Plus · Published 2026-07-07 · DOI 10.1016/j.phyplu.2026.101023
Phateep Hankittichai, Narudol Teerapattarakan, Shisanupong Anukanon
Objective: To identify and experimentally validate anti-inflammatory phytochemicals from Thai medicinal plants and to examine the relationship between in silico target prediction, enzymatic inhibition, and cellular activity. Methods: A systematic review was performed to identify Thai medicinal plants with documented traditional applications in the treatment of inflammatory conditions. Phytochemicals identified in the selected species were assessed via molecular docking simulations against critical enzymes involved in inflammation, specifically cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), xanthine oxidase (XOD), and inducible nitric oxide synthase (iNOS). Subsequent analyses of the selected compounds encompassed enzyme inhibition assays, molecular dynamics simulations, and cellular assays utilizing lipopolysaccharide-stimulated RAW 264.7 macrophages. Network pharmacology analysis was subsequently performed to explore pathway-level mechanisms associated with observed cellular effects. Results: Triptocalline A (TRP) and daphnoretin (DAP) were prioritized based on docking profiles and target coverage. Despite favorable predicted binding affinities, both compounds exhibited weak direct enzyme inhibition. Molecular dynamics simulations revealed target-dependent interaction stability but did not fully account for biochemical outcomes. In contrast, TRP demonstrated significant cellular anti-inflammatory activity by reducing nitric oxide (NO) production and suppressing iNOS expression at non-cytotoxic concentrations, whereas DAP showed limited effects. Network pharmacology analysis revealed that TRP-associated targets were enriched in immune- and cytokine-related signaling processes, including pathways linked to MAPK activation and interleukin-mediated responses, supporting a role for pathway-level modulation. Conclusions: This study demonstrates a clear disconnect between docking-predicted affinity and functional biological activity for complex phytochemicals. The findings highlight that anti-inflammatory effects of TRP are primarily mediated through pathway-level regulation, particularly involving the iNOS–NO axis, rather than direct enzymatic inhibition. Integrating computational, biochemical, cellular, and network-based analyses provides a more reliable framework for interpreting phytochemical activity and prioritizing bioactive compounds from traditional medicinal sources.
Abstract from DOAJ. Public domain (CC0 1.0).
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Hankittichai, P., Teerapattarakan, N., Anukanon, S. (2026). From in silico prediction to cellular validation: Discrepancies between docking affinity and biological activity of anti-inflammatory phytochemicals from Thai medicinal plants. Phytomedicine Plus. https://doi.org/10.1016/j.phyplu.2026.101023