Exogenous phosphocreatine (PCr) and its cyclic analog, 3-phosphono-2-imino-1-methyl-4-oxoimidazolidine (PIMOI) were used as protectors of ischemic myocardium. PCr was insignificantly metabolized, whereas its analog was rapidly split, resulting in the formation of creatinine and inorganic phosphate as well as of minor amounts of PCr and creatine. Exogenous AMP and ATP accelerated PCr breakdown; in contrast, PIMOI hydrolysis slowed down in the presence of AMP. A similar inhibitory effect was observed after treatment of hearts with 2,4-dinitrofluorobenzene. These data together with those obtained for heart homogenates point to the enzymatic nature of PIMOI hydrolysis in ischemic heart. Acidic phosphatases and 5'-nucleotidases of the heart are supposed to be involved in this process.