Anti-inflammatory glycoside withanolides from leaves of Athenaea velutina

The phytochemical study of the CH₂Cl₂ and EtOAc soluble fractions from the leaf ethanol extract of Athenaea velutina resulted in the isolation of the new withanolides 1-3, two of which substituted at the C-3 by a disaccharide 3β-O-[α-L-rhamnopyranosyl-(1 → 4)-β-d-glucopyranosyl], and four others previously reported (4-7). The structures of the new withanolides were elucidated by detailed interpretation of their 1D- and 2D-NMR spectra and the HRESIMS analysis. An anti-inflammatory screening was performed using the two major withanolides (1 and 2), analyzing their effects on the viability of RAW 264.7 cells and their ability to inhibit cellular nitric oxide production. While withanolide 1 exhibited pro-inflammatory activity by increasing nitrite production in LPS-stimulated macrophages, compound 2, in opposition, demonstrated anti-inflammatory action by reducing those levels.