One new isoflavone, 6′-dihydroxy-2-methyl-2′-methoxyl-3′-acetylisoflavone (1) and six known compounds (2−7) were isolated from the processed root barks of Paeonia suffruticosa. The structures of these compounds were elucidated using UV, IR, NMR, and HR-ESI-MS data. Compound 1 represents the first example of naturally occurring 2-substituted isoflavone. Compounds 2−7 were isolated from the processed root barks of P. suffruticosa for the first time. In addition, all of the compounds were screened for their procoagulant activity. As a result, compounds 4, 6, and 7 showed significantly in vitro procoagulant activity.