Development of specific bioluminescent in vitro assays for selecting potential antimineralocorticoids.
Efficient antimineralocorticoid selection requires a reliable, discriminating and easy assay for monitoring biological activity of not only the specific receptor, but also closely related receptors such as glucocorticoid and progestin. These related activities should be as low as possible to obtain specific antimineralocorticoid compounds. In this paper, we describe two cellular models used for easy and specific measurement of mineralocorticoid and progestin activities. These models involve the induction of firefly luciferase under hormonal control mediated by a chimeric receptor. The first model comprises transiently transfected MCF-7 cells, whereas the second uses stably transfected HeLa cells. Glucocorticoid activity was assayed with the classic tyrosine-aminotransferase induction method in HTC cells. Six compounds of a new family of 11 beta-substituted-17-spirolactone steroids were thus studied and compared to control compounds. Five of them showed antimineralocorticoid activity and one was active at a concentration lower than that of mespirenone.
Jausons-Loffreda
N
N
INSERM U58, Montpellier, France.
Balaguer
P
P
Auzou
G
G
Pons
M
M
D000818:Animals; D001483:Base Sequence; D001681:Biological Assay / Q000379:methods*; D002460:Cell Line; D003907:Dexamethasone / Q000378:metabolism; D006367:HeLa Cells; D006727:Hormone Antagonists / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis*; D008163:Luminescent Measurements; D015735:Mifepristone / Q000378:metabolism; D000451:Mineralocorticoid Receptor Antagonists / Q000378:metabolism / Q000494:pharmacology; D008901:Mineralocorticoids / Q000378:metabolism*; D008969:Molecular Sequence Data; D011282:Pregnenediones / Q000378:metabolism; D011397:Promegestone / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D018161:Receptors, Mineralocorticoid / Q000235:genetics / Q000378:metabolism*; D011980:Receptors, Progesterone / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism / Q000494:pharmacology; D013148:Spironolactone / Q000031:analogs & derivatives / Q000378:metabolism; D013739:Testosterone / Q000378:metabolism; D014162:Transfection
D006727:Hormone Antagonists; D000451:Mineralocorticoid Receptor Antagonists; D008901:Mineralocorticoids; D011282:Pregnenediones; D011965:Receptors, Glucocorticoid; D018161:Receptors, Mineralocorticoid; D011980:Receptors, Progesterone; D011993:Recombinant Fusion Proteins; D013148:Spironolactone; D015735:Mifepristone; D013739:Testosterone; D003907:Dexamethasone; D011397:Promegestone; D008156:Luciferases