Two new isoflavone glycosides, tectorigenin 7-O-beta-D-glucopyranoside-4'-O-[beta-D-glucopyranosyl-(1''''-->6''')-beta-D-glucopyranoside] (1) and iristectorigenin B 4'-O-[beta-D-glucopyranosyl-(1'' --> 6'')-beta-D-glucopyranoside] (2), together with 11 known compounds, including six isoflavones, tectorigenin 7-O-beta-D-glucopyranoside-4'-O-beta-D-glucopyranoside (3), tectorigenin 4'-O-[beta-D-glucopyranosyl-(1'''--> 6'')-beta-D-glucopyranoside] (4), tectorigenin 7-O-beta-D-glucopyranoside (5), genistein 7-O-beta-D-glucopyranoside (6), tectorigenin 4'-O-beta-D-glucopyranoside (7), and tectorigenin (8); two phenolic acid glycosides, vanillic acid 4-O-beta-D-glucopyranoside (9) and glucosyringic acid (10); a phenylpropanoid glycoside, E-coniferin (11); an auronol derivative, maesopsin 6-O-beta-D-glucopyranoside (12); and a pyrrole derivative, 4-(2-formyl-5-hydroxymethylpyrrol-1-yl) butyric acid (13), were isolated from fresh Iris spuria (Calizona) rhizomes. The structures of these compounds were established on the basis of spectroscopic and chemical evidence. Inhibitory effects on the activation of Epstein-Barr virus early antigen were examined for compounds 1-8 and 12.