title: Novel antiprogestins Org 31806 and 31710: interaction with mammalian progesterone receptor and DNA binding of antisteroid receptor complexes.
abstract: We have examined steroid binding parameters and transformation of calf uterine progesterone receptor (PR) liganded with progestins (progesterone and R5020) and the newly synthesized antiprogestins (Org 31806 and 31710). Species specificity analysis indicated that [3H]R5020 binding in the chicken oviduct cytosol could be eliminated in the presence of 100-fold excess radioinert progesterone and R5020 but not Org 31806 and 31710. In the calf uterine cytosol, the progestins and the antiprogestins appeared to interact with the same PR as revealed by the displacement of [3H]R5020 by all of the above steroids. When the extent of [3H]R5020 binding was examined in the presence of different concentrations of radioinert steroids, the relative affinity with which these compounds interacted with the uterine PR was found to be comparable. A 23 degrees C incubation of cytosol transformed the progestin-bound PR complexes increasing their binding to DNA-cellulose from 5 (0 degrees C, nontransformed) to 35%. Under these conditions, 20% Org 31710- and RU486-occupied PR complexes bound to DNA-cellulose whereas only 10% Org 31806-receptor complexes were retained by the resin. Transformation (23 degrees C) of cytosol receptor caused a loss of the larger 8 S form and an increase in the smaller 4 S form. In its unliganded state or when it was complexed with R5020 or the antiprogestins, incubation of PR at 23 degrees C led to dissociation of the receptor-associated 90 kDa heat-shock protein (hsp90). The PR-hsp90 association was stabilized in the presence of 10 mM iodoacetamide when the ligand binding site was occupied by Org 31806 and 31710. The R5020-receptor complexes, however, allowed release of hsp90 under the above transforming conditions. Our results indicate that although Org 31806 and 31710 show no affinity for the avian PR, these steroids interact with the mammalian PR. We propose that the reported antiprogestational effects of Org 31806 and 31710 are mediated via their interaction with PR which appears similar to one that exists between PR and RU486.
authors.affiliation: Department of Biological Sciences, Oakland University, Rochester, MI 48309-4401.
authors.forename: T
authors.initials: T
authors.lastname: Mizutani
authors.forename: A
authors.initials: A
authors.lastname: Bhakta
authors.forename: H J
authors.initials: HJ
authors.lastname: Kloosterboer
authors.forename: V K
authors.initials: VK
authors.lastname: Moudgil
mesh_terms: D000818:Animals; D002417:Cattle; D002645:Chickens; D003600:Cytosol / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D004963:Estrenes / Q000494:pharmacology*; D005260:Female; D005663:Furans / Q000494:pharmacology*; D007700:Kinetics; D015735:Mifepristone / Q000494:pharmacology; D011374:Progesterone / Q000494:pharmacology; D011372:Progestins / Q000037:antagonists & inhibitors*; D011397:Promegestone / Q000494:pharmacology; D011980:Receptors, Progesterone / Q000187:drug effects* / Q000378:metabolism; D013045:Species Specificity; D014599:Uterus / Q000166:cytology
chemical_list: D004963:Estrenes; D005663:Furans; D011372:Progestins; D011980:Receptors, Progesterone; C076196:Org 31710; C076197:Org 31806; D015735:Mifepristone; D011374:Progesterone; D004247:DNA; D011397:Promegestone