Two new glycosylated piericidins, glucopiericidinol A3 (1) and 7-demethyl-glucopiericidin A (2), along with four known analogs were isolated from the culture broth of Streptomyces sp. KIB-H1083. The chemical structures of new compounds were elucidated by spectroscopic analyses. Their cytotoxicity on HL-60, SMMC-772, A-549, MCF-7, and SW480 cell lines, as well as antimicrobial activities was evaluated. The results showed that glucopiericidin A (4) has potent cytotoxicity against HL-60, SMMC-772, A-549, and MCF-7 cell lines with IC50 values of 0.34, 0.65, 0.60, and 0.50 μM, respectively. For the antimicrobial activity, piericidin A (6) showed most powerful inhibitory activities against Xanthomonas oryzae pv. oryzicola, and Penicillium decumbens.