Abstract
We found previously that stimulation of natriuretic peptide receptor (NPR)-B by C-type natriuretic peptide (CNP) in failing rat ventricle potentiates β1-adrenoceptor (β1-AR)-mediated inotropic response to noradrenaline through cGMP-mediated inhibition of phosphodiesterase (PDE) 3, thereby enhancing cAMP-mediated signalling. Increased cAMP-mediated signalling is deleterious in chronic heart failure (HF; basis for the use of β-blockers in HF) and we propose to consider NPR-B antagonists as new HF treatment in addition to conventional therapy. Since there is no NPR-B-selective antagonist available for clinical studies, we aimed at identifying a novel small molecule (non-peptide) NPR-B antagonist. An assay was developed and high throughput screening performed on a chemical library of about 20,000 small molecule compounds (<500 Da) to identify NPR-B antagonists based on inhibition of CNP-stimulated cGMP production in NPR-B-expressing HEK293 cells. The screen revealed several potential NPR-B antagonists, of which six were selected for further studies. Three showed selective NPR-B vs NPR-A inhibition and three were partially selective. The compounds mediated reversible, noncompetitive inhibition and most likely act as allosteric modulators binding outside the agonist binding site of NPR-B. In rat ventricular muscle strips, the potentiating effect of CNP upon β1-AR-evoked inotropic effects could be attenuated by at least one of these compounds. We identified several small molecule NPR-B antagonists by high throughput screening and show in a functional heart preparation that blocking NPR-B stimulation with a small molecule compound can reduce the potentiating effect of CNP on the β1-AR-mediated inotropic response to noradrenaline.
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Acknowledgments
We wish to thank Niko Sahlberg and Prof. Kjetil Taskén at the Biotechnology Centre of Oslo for excellent help and support with the screening of the compound libraries. This work was supported by The Norwegian Council on Cardiovascular Disease, The Research Council of Norway, The Kristian Gerhard Jebsen Foundation, Anders Jahre’s Foundation for the Promotion of Science, The Family Blix Foundation, The Simon Fougner Hartmann Family Foundation, South-Eastern Norway Regional Health Authority, University of Oslo and the COST Action BM1005 (European Network on Gasotransmitters).
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Bach, T., Bergholtz, S., Riise, J. et al. Identification of small molecule NPR-B antagonists by high throughput screening — potential use in heart failure. Naunyn-Schmiedeberg's Arch Pharmacol 387, 5–14 (2014). https://doi.org/10.1007/s00210-013-0940-6
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DOI: https://doi.org/10.1007/s00210-013-0940-6