Five constituents were extracted from the aerial part of Paederia pertomentosa and isolated by column chromatography. Their structures were elucidated by physicochemical properties and spectroscopic data analysis. The isolated compounds were identified as 1,2-dimethoxy-3,4-dihydroxyanthraquinone named as paederone (1), paederoside (2), deacetyl asperulosidic acid methyl ester (3), paederosidic acid (4) and methylpaederosidate (5). Compound 1 is a new compound which exhibits a significant inhibitory effect on Escherichia coli and Staphylococcus aureus. Compounds 2-5 were isolated from this plant for the first time.