A new mode of mineralocorticoid receptor antagonism by a potent and selective nonsteroidal molecule

J Biol Chem. 2010 Sep 24;285(39):29932-40. doi: 10.1074/jbc.M110.131342. Epub 2010 Jul 22.

Abstract

Limitations of current steroidal mineralocorticoid receptor (MR) antagonists have stimulated the search for a new generation of molecules. We screened for novel nonsteroidal compounds and identified MR antagonists derived from the chemical class of dihydropyridines. Chemical optimization resulted in BR-4628, which displays high in vitro and in vivo MR potency as well as selectivity with respect to the other steroid hormone receptors and the L-type calcium channel. Biochemical studies demonstrated that BR-4628 forms complexes with MR that do not promote the recruitment of transcriptional co-regulators. Docking experiments, using the crystal structure of the MR ligand-binding domain in an agonist conformation, revealed that BR-4628 accommodates in the MR ligand-binding cavity differently in comparison with the classical steroidal MR antagonists. An alanine scanning mutagenesis approach, based on BR-4628 docking, allowed identifying its anchoring mode within the ligand-binding cavity. Altogether, we propose that BR-4628 is a bulky antagonist that inactivates MR through a passive mechanism. It represents the prototype of a new class of MR antagonists.

MeSH terms

  • Amino Acid Substitution
  • Animals
  • Binding Sites
  • CHO Cells
  • Calcium Channels, L-Type / genetics
  • Calcium Channels, L-Type / metabolism
  • Cricetinae
  • Cricetulus
  • Crystallography, X-Ray
  • Dihydropyridines / chemistry
  • Dihydropyridines / pharmacology*
  • Drug Evaluation, Preclinical
  • Humans
  • Ligands
  • Mineralocorticoid Receptor Antagonists*
  • Mutation, Missense
  • Receptors, Mineralocorticoid / genetics
  • Receptors, Mineralocorticoid / metabolism

Substances

  • Calcium Channels, L-Type
  • Dihydropyridines
  • Ligands
  • Mineralocorticoid Receptor Antagonists
  • Receptors, Mineralocorticoid