Inhibition of ABCC5-mediated cGMP transport by progesterone, testosterone and their analogues

J Steroid Biochem Mol Biol. 2021 Oct:213:105951. doi: 10.1016/j.jsbmb.2021.105951. Epub 2021 Jul 13.


The biodynamics and biokinetics of sex hormones are complex. In addition to the classical steroid receptors (nuclear receptors), these hormones act through several non-genomic mechanisms. Modulation of ABC-transporters by progesterone represents a non-genomic mechanism. In the present study, we employed inside out vesicles from human erythrocytes to characterize high affinity cGMP transport by ABCC5 (member 5 of the ATP-Binding Cassette subfamily C). Progesterone and testosterone inhibited the transport with respective Ki of 1.2 ± 0.3 and 2.0 ± 0.6 μmol/L. We used virtual ligand screening (VLS) to identify analogues to progesterone and testosterone. A large number of substances were screened in silico and the 19 most promising candidates were screened in vitro. Each substance was tested for a concentration of 10 μmol/L. The range of cGMP transport reduction was 21.5% to 86.2% for progesterone analogues and 8.6% to 93.8 % for testosterone analogues. Three of the most potent test compounds (TC) of each analogue class, in addition to progesterone and testosterone, were characterized for concentrations from 1 nanomol/L to 1 mmol/L. The progesterone analogues showed following Ki-values (μmol/L): TC-08: 0.61, TC-16: 0.66 and TC-15: 9.3. The Ki-values (μmol/L) for the testosterone analogues were: TC-18: 0.10, TC-07: 0.67 andTC-05: 2.0. The present study shows that VLS may be a versatile tool in the development of membrane transport modulating agents (MTMAs).

Keywords: ABCC5; Analogues; Cellular efflux; Cyclic GMP; MRP5; Progesterone; Testosterone; Transport modulation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Transport / drug effects
  • Cyclic GMP / metabolism*
  • Dose-Response Relationship, Drug
  • Erythrocyte Membrane / drug effects*
  • Erythrocyte Membrane / metabolism
  • Gene Expression
  • High-Throughput Screening Assays
  • Humans
  • Kinetics
  • Ligands
  • Multidrug Resistance-Associated Proteins / antagonists & inhibitors
  • Multidrug Resistance-Associated Proteins / genetics
  • Multidrug Resistance-Associated Proteins / metabolism*
  • Progesterone / analogs & derivatives
  • Progesterone / pharmacology*
  • Protein Binding
  • Structure-Activity Relationship
  • Testosterone / analogs & derivatives
  • Testosterone / pharmacology*
  • User-Computer Interface


  • ABCC5 protein, human
  • Ligands
  • Multidrug Resistance-Associated Proteins
  • Testosterone
  • Progesterone
  • Cyclic GMP