ETHNOPHARMACOLOGICAL RELEVANCE: Pueraria montana var. lobata (PM) has the effects of relieving muscle stiffness and fever, generating body fluids and quenching thirst, resolving rashes, raising yang and stopping diarrhea, unblocking meridians, and detoxifying alcohol. It is commonly used for the management of conditions including stiff neck and back pain, thirst, diabetes, unresolved measles, external fever with headache, dysentery, diarrhea, dizziness and headache, stroke with hemiplegia, chest and heart pain, and alcohol poisoning. However, research on the material basis and mechanism of action of its anti-inflammatory efficacy is still quite lacking. AIM OF THE STUDY: The objective is to look into the inflammation-dampening characteristics of PM through in vitro studies and to accurately identify the phytochemicals within the therapeutic herb that correlate with its customary applications. MATERIALS AND METHODS: A comprehensive phytochemical investigation was carried out using chromatographic and spectral techniques to explore the constituents of PM. Potential targets of the active chemical that might reduce inflammation were predicted using network pharmacology. The inhibition of inflammatory mediators in RAW264.7 cells stimulated by lipopolysaccharide (LPS) was used to measure the anti-inflammatory effects in vitro. RESULTS: The research revealed that the PM chloroform extract exhibited significant anti-inflammatory action by efficiently preventing NO release in LPS-activated RAW264.7 cells. Further phytochemical analysis led to the identification and characterization of 29 natural products, including 4 new compounds (1-4). Among these, compounds 1, 4, 7, 9-18, and 20-25 exhibited significant anti-inflammatory activity, with compound 1 showing the most potent effect. Subsequent network pharmacology, along with molecular docking and molecular dynamics simulations, suggested that 1 targets several key anti-inflammatory pathways, including HSP90AA1, MAPK, mTOR, and NF-κB. In vitro validation confirmed that the mechanism of anti-inflammation of 1 involves modulation of the HSP90AA1/MAPK/mTOR/NF-κB signaling pathways. CONCLUSIONS: This study not only more or less supports the traditional use of PM for its anti-inflammatory properties but also introduces novel pterocarpan-type isoflavone as promising agent for inflammation management.