Guide isolation of apotirucallane and tirucallane triterpenoids from the branches and leaves of Brucea javanica (L.) Merr. and their anti-inflammatory activities.

In this study, the chemical constituents of the branches and leaves of Brucea javanica were investigated using an integrated approach combining molecular networking, the Moldiscovery strategy, and traditional phytochemical techniques. This strategy led to the isolation and characterization of nine previously undescribed triterpenoids, named brujavaneos A-I (1-9). Their structures were unambiguously established through extensive spectroscopic analysis and electronic circular dichroism calculations. Furthermore, the anti-inflammatory activities of these isolates were evaluated. Several compounds exhibited moderate activity, with their IC50 values reliably predicted via InflamNat, a machine-learning-based tool. Notably, compound 4 demonstrated significant inhibition of nitric oxide production in LPS-stimulated RAW264.7 macrophages, with an IC50 value of 15.03 ± 0.03 μM.