Chemical investigation of Chrozophora tinctoria (L.) A. Juss. growing in Saudi Arabia revealed the isolation of two new acylated flavonoids identified as acacetin-7-O-b-D-[a-L-rhamnosyl(1!6)]300-E-p-coumaroyl glucopyranoside (4) and apigenin-7-O-(600-Z-p-coumaroyl)-b-D-glucopyranoside (5), in addition to amentoflavone (1), apigenin-7-O-b-D-glucopyranoside (2), apigenin-7-O-600-E-p-coumaroyl-b-D-glucopyranoside (3) and rutin (6). The structures of isolated compounds were established by 1D, 2D NMR and HRESIMS spectral data, in addition to comparison with literature data. The anti-inflammatory activities of isolated compounds were assessed by measuring the levels of IL-1b, IL-6, TNF-a and PGE2 in the supernatant media of human peripheral blood mononuclear cells (PBMCs) stimulated by phytohaemagglutinin (PHA). At a concentration of 100mM, compounds 1, 2, 4 and 6 significantly decreased Il-1b, Il-6 and PGE2 to nearly normal values. All tested compounds caused a dose-dependent decrease in TNF-a level but failed to reach that of the control values.