Nitric Oxide Production Inhibitory Terpenylated Coumarins from Ailanthus altissima

LC-MS/MS-based molecular networking was used to guide the targeted isolation of terpenylated coumarins from the barks of Ailanthus altissima. Five known terpenylated coumarins (1–5), along with two simple coumarins (6 and 7), were isolated from the CH₂Cl₂-soluble fraction. Their structures were determined using spectroscopic techniques, including 1D and 2D NMR and LC-HR-MS/MS. Notably, altissimacoumarins C (1), F ( 3), and H (4) demonstrated potent inhibition of LPS-induced nitric oxide production in RAW 264.7 macrophages, with IC₅₀ values of 36.1, 27.9, and 11.2 μM, respectively. These results demonstrate the anti- inflammatory properties of A. altissima, supporting its possible application in treating inflammatory disorders.