Interaction of estrogen-nitrosourea conjugates with the estrogen receptor in rat uterus.

Several estrogen-nitrosourea conjugates have been synthesized with the aim of producing more selective cytotoxic agents. The conjugates were shown to compete with estradiol for binding to cytosolic estrogen receptor in rat uterus; the relative binding affinities for N-(2-chloroethyl)-N'-(3,17 beta-dihydroxyestra-1,3,5(10)-trien-17 alpha-yl)methyl-N-nitrosourea (17 alpha-CNU), N-(2-chloroethyl)-N'-(3-hydroxyestra-1,3,5(10)-trien-17 beta-yl)-N-nitrosourea (17 beta-CNU), and N-(2-chloroethyl)-N'-2,3-di(p-hydroxyphenyl)-pentanyl-N-nitrosoure a (HEX-CNU) were 2, 0.4, and 0.2, respectively, using a binding affinity of 100 for estradiol. In the ligand exchange assay, cytosolic receptors preloaded with 17 alpha-CNU and HEX-CNU were found to lose some of their estradiol (E2) binding sites, suggesting that binding to estrogen receptor (ER) may be irreversible. An increase of nuclear accumulation of ER was observed in the presence of 17 alpha-CNU and HEX-CNU. In the rat system, even at a 10,000-fold excess, these two agents failed to show any antagonism of the uterotrophic effect of E2 in vivo. The low binding affinity and instability of these conjugates may account for their lack of antiestrogen activity. On the other hand, 17 alpha-CNU at 100 or 1000 micrograms/day and HEX-CNU at 1000 micrograms/day demonstrated significant uterotrophic activity. This study did not resolve whether the stimulation of uterine growth was due to the parent estrogen-nitrosourea conjugate or to decomposition and/or metabolic products.