In vivo and in vitro estrogenic profile of 17β-amino-1,3,5(10)estratrien-3-ol

• We analyzed the estrogenic profile of 17β-amino-1,3,5(10)estratrien-3-ol (17βAE2).
• 17βAE2 is a weak estrogenic agonist with actions mediated mostly by ERβ.
• 17αAE2 binds with lower relative affinity than 17βAE2 to hERα and hERβ.
• Change of NH2 instead of the OH in C17β of 17βE2 decrease its estrogenic activity.

17β-amino-1,3,5(10)estratrien-3-ol (17βAE2), is the 17β-aminoestrogens prototype possessing anticoagulant activity, contrasting with the procoagulant effects of 17β-estradiol (17βE2). Its estrogenicity profile has not been reported, and it was evaluated by uterotrophic assay, estrogen receptor binding affinity and its ability to induce gene transcription of the human estrogen receptor (hER)α mediated in a Saccharomyces cerevisiae yeast expression system. Additionally, 17βAE2 and 17αAE2 were compared with 17βE2 in HeLa cells co-transfected with expression vectors for hERα or hERβ subtypes and for an estrogen-responsive reporter gene. Immature female CD1 mice and Wistar rats (21 days old) were treated for three days with 17βAE2 (10–5000 μg/kg), 17βE2 (0.001–1000 μg/kg) or vehicle (propylenglycol 10 ml/kg) and uterine weights were estimated. 17βAE2 increased uterine weight in a dose-dependent manner. The effective dose (ED)50 uterine weight values: 17βAE2 = 552 and 764 μg/kg (17βE2 = 4.8 and 16 μg/kg) and their relative uterotrophic potency were 0.86 and 2.1 (17βE2 = 100) in mice and rats, respectively. 17βAE2 competed with [3H]E2 for the estrogen receptor. The 17βAE2 relative binding affinities (RBAs) were: 0.074; Ki = 2.2 × 10−6 M (17βE2 = 100; Ki = 1.6 × 10−9 M); 0.029 and Ki = 3.8 × 10−6 M (17βE2 = 100; Ki = 1.1 × 10−9 M) for mice and rats uteri respectively. 17βAE2 activated hERα-mediated β-galactosidase transcription activity in the yeast system co-transfected with hERα gene. 17βAE2 effective concentration (EC)50 = 1.82 μM (17βE2 = 2.14 nM) with a relative potency of 0.12 (17βE2 = 100). These transactivation effects were abolished by the antagonist fulvestrant (ICI 182,780), similarly to 17βE2. 17βAE2 and 17αAE2 bind with low relative affinity to hERα and hERβ. Both induced hER-mediated reporter gene transactivation in a dose-response manner. The overall results provide evidence that 17βAE2 has a weak agonist estrogenic action greatly mediated through the hERβ and to a lesser extent the hERα at genomic level.