Modulation of estrogen receptor transactivation and estrogen-induced gene expression by ormeloxifene-a...

The study was aimed to investigate the interaction of D,L-ormeloxifene (Orm), a triphenylethylene and its hydroxy derivative with estrogen receptor subtypes alpha and beta, its influence on ERE-driven transcriptional activation and progesterone receptor expression. In competitive binding experiments using human recombinant ERalpha and ERbeta, Orm showed interaction with both ER subtypes, with more selectivity and higher affinity towards ERalpha (8.8%) as compared to ERbeta (3%). In case of 7-hydroxy derivative, the relative binding affinity for both ERs was enhanced several folds. Orm showed lower Ki, i.e. higher affinity for ERalpha (250 nM) than for ERbeta (750 nM). It was observed that Orm promoted the formation of ER-ERE complexes in uterine tissue extract whereas its hydroxy derivative showed inhibitory effects. Transient co-transfection assay in COS-1 cells using ERE-luciferase reporter construct, revealed that Orm showed estrogenic response whereas its hydroxy-derivative was potent antiestrogen at ERalpha at transcription level. In immature rats, Orm (2 mg/kg) was associated with less increase in uterine weight and in luminal epithelial cell height than E2 or Tam. Orm also induced the expression of PR mRNA but the expression level was significantly less than estradiol treated group. These results suggest that ER-ERE complexes formed under the influence of 7-hydroxy Orm appear to be transcriptionally less effective hence antagonizing the E2-regulated gene expression in this target tissue.