Differences in the determinants of eplerenone, spironolactone and aldosterone binding to the mineralocorticoid receptor

Summary

1. The importance of mineralocorticoid receptor (MR) antagonists in the treatment of cardiovascular disease has been emphasised by two recent clinical trials, one using spironolactone and the other using a new selective MR antagonist, namely eplerenone.

2. Eplerenone has a very low affinity for the glucocorticoid receptor (GR). Determinants of binding specificity of eplerenone to the MR were investigated using chimeras created between the ligand-binding domains (LBD) of the MR and the GR. These chimeras had been used previously to investigate aldosterone and spironolactone binding specificity to the MR.

3. Eplerenone competed strongly for [³H]-dexamethasone binding to a MR/GR chimera containing amino acids 804–874 of the MR and weakly to a chimera containing amino acids 672–803 of the MR. Within the 804–874 region, eplerenone competed for [³H]-dexamethasone binding to a chimera containing amino acids 820–844 of the MR, although the calculated affinity was approximately 10-fold lower than for binding to the full-length MR LBD. Similar results were obtained using another MR antagonist, namely spironolactone. Modelling of eplerenone binding to the MR LBD, based on the GR LBD crystal structure, suggests that amino acids 820–844 affect the overall shape of the ligand-binding pocket and that eplerenone acts as an MR antagonist because it fails to stabilize the active conformation of the receptor.

4. In contrast with results with the MR antagonists eplerenone and spironolactone, amino acids 820–844 are sufficient in themselves to confer high-affinity aldosterone binding to the MR, suggesting that the binding determinants of the two antagonists are similar to each other but differ from those of aldosterone.