Eleven healthy male volunteers, ages 25-39 years, received a single dose of synthetic delta sleep-inducing peptide (DSIP) (25 nmol/kg BW) or saline intravenously in a randomized cross-over, double-blind study. The concentrations of neuropeptides related to the hypothalamic pituitary-adrenal (HPA) axis and cortisol were examined in serial plasma samples. In addition, cortisol and monoamine metabolites were determined in urine. A significant reduction of ACTH-like immunoreactivity (ACTH-LI) in plasma was detected for at least 3 hr after the DSIP injection, compared to the control subjects, in whom a slightly elevated concentration of ACTH-LI occurred. Plasma cortisol levels were unaffected and followed the normal diurnal decline. No differences in urinary cortisol or monoamine metabolite concentrations occurred between the two groups. The results indicate an inhibitory action of DSIP on ACTH secretion in man, as previously suggested by animal experiments.