EVALUATION OF THE DISODIUM SALT OF 4,4'-DIAMINO-2,2'-STILBENE DISULFONIC ACID FOR ESTROGENIC ACTIVITY

4,4'-Diamino-2,2'-stilbene disulfonic acid (DAS), a key intermediate i n the synthesis oi dyes and fluorescent whitening agents, has been pur ported to have weak estrogenic properties based on apparent structural similarity with diethylstilbestrol (DES) and unsubstantiated reports of male reproductive dysfunction in an industrial setting. In weanling rats, high doses of DAS (300 mg/kg ip; 1000-3000 mg/kg oral) have bee n associated with modest increases in the uterus/body weight ratio (Sm ith & Quinn, 1992). In order to more directly and definitively determi ne ii DAS possesses estrogenic activity, in vitro studies were conduct ed to establish its relative binding affinity to the human estrogen re ceptor (ER) in MCF-7 cells, a well-characterized breast cancer cell li ne. At concentrations approaching its solubility limit (10(-4) M), DAS failed to displace [-H-3]-17 beta-estradiol (E(2)) from the ER. In co ntrast, DES and E(2) demonstrated characteristic competitive binding c urves (50% displacement of H-3-E(2) at 3.33 x 10(-9) and 1.33 x 10(-8) M, respectively). Parallel in vivo comparisons of DAS (10 or 30 mg/an imal) and DES (1 or 3 mu g/animal) were also conducted to assess utero tropic effects. After three daily subcutaneous injections, DAS did not induce uterine weight gain. In contrast DES consistently and markedly increased uterine weight and induced uterine water imbibition, with t he latter effect being absent in DAS-treated rats. Under these experim ental conditions, DAS was shown to possess negligible, ii any, estroge nic activity, despite apparent structural similarity with known estrog ens.