Benjalokawichian (BLW) is a classic antipyretic polyherbal remedy used in Thai traditional medicine (TTM) to reduce toxic fever (TF). This study aimed to shed light on the mechanisms of action and identify bioactive components of BLW responsible for TF treatment. The methods that combine network pharmacology, molecular docking, and the inhibition of nitric oxide (NO) production in LPS-induced RAW 264.7 were employed for these objectives. Network pharmacology served as a means to identify 15 potential bioactive compounds, 88 possible therapeutic targets, and 4 hub genes related to BLW. Among the significant targets, TNF, PTGS2, STAT3, and NFKB1 were closely linked to the metabolic pathways of phenylalanine, arachidonic acid, and tyrosine, which are vital in managing infections, inflammation, proliferation, and apoptosis in the TF microenvironment. Additionally, molecular docking analysis indicated that core compounds displayed strong binding affinities for the key targets, with binding energies ranging between −4.5 and −11.1 kcal/mol. The in vitro assay demonstrated that BLW extract significantly inhibited NO production in LPS-activated RAW 264.7 macrophages, presenting an IC50 value of 69.10 μg/mL, and no cytotoxic effects on RAW 264.7 macrophages. Furthermore, the biomarker compounds of BLW extract, viz., perforatic acid and peucenin-7-methyl ether were found to decrease NO production in a dose-dependent manner. In summary, this research indicates that BLW provides therapeutic benefits for TF via a complex interplay of different compounds, targets, and pathways. These findings serve as a foundation for further research into the mechanisms of action of a polyherbal remedy toward TF to provide scientific evidences for its clinical use.
Chaloemram et al. (Mon,) studied this question.