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Synthesis and characterisation of NS13558: a new important tool for addressing KCa1.1 channel function ex vivo

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Abstract

Pharmacological activation of the large-conductance Ca2+-activated K+ channel (KCa1.1) in the cardiac inner mitochondrial membrane has been found to protect the heart against ischemia reperfusion injuries. However, there are concerns about the selectivity of the pharmacological tools used to modulate the channel. Here, we address this issue by synthesising a methylated analogue of the tool KCa1.1 channel activator NS11021. The compound (NS13558) is designed as a structurally closely related and biologically inactive analogue of NS11021. NS13558 did not elicit any significant opening of cloned human KCa1.1 channels, but maintained comparable biological activity towards other cardiac ion channels as compared to NS11021. In isolated perfused rat hearts subjected to ischemia–reperfusion, infarct size was reduced from 29% in control to 7% in NS11021 treated hearts. In comparison, the inactive derivate of NS11021, i.e., NS13558, did not confer any cardioprotection, demonstrated by an infarct size identical to control hearts. This suggests that NS11021 exerts its primary effect through KCa1.1 channels, which indicates an important role of these channels in protection against ischemia–reperfusion injuries. Furthermore, the study demonstrates a novel way of combining an activator of the KCa1.1 channel (NS11021) and its structurally closely related inactive analogue NS13558 to address the functional role of KCa1.1 channels, and we believe these novel tools may constitute a valuable addition to understanding the functional role of KCa1.1 channels under physiological and pathophysiological conditions.

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Abbreviations

HEK-cells:

human embryonic kidney 293 cells

DMSO:

dimethyl sulphoxide

LVDP:

Left ventricular developed pressure

LVeDP:

left ventricular end-diastolic pressure

TTC:

2,3,5-triphenyltetrazolium chloride

APD:

action potential duration at 90% repolarization

UPLC:

ultra performance liquid chromatography

MS:

mass spectrometry

FT-IR:

Fourier transform infrared spectroscopy

HPLC:

high-performance liquid chromatography

ESI-HRMS:

electron spray ionisation high resolution mass spectra

LC:

liquid chromatography

M.p.:

melting point

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Acknowledgements

We thank Camilla Irlind for excellent technical assistance. This study was supported by the Novo Nordisk Foundation, the Aase and Ejnar Danielsen Foundation, the Danish National Research Foundation Centre for Cardiac Arrhythmia, and the Danish Medical Research Council.

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Authors and Affiliations

  1. Danish National Research Foundation Centre for Cardiac Arrhythmia, University of Copenhagen, Blegdamsvej 3, 2200, Copenhagen, Denmark

    Bo Hjorth Bentzen & Søren-Peter Olesen

  2. Department of Medicinal Chemistry, NeuroSearch A/S, Pederstrupvej 93, 2750, Ballerup, Denmark

    Rune Wederkinck Andersen & Antonio Nardi

  3. NeuroSearch A/S, Drug Discovery Portfolio Management, Pederstrupvej 93, 2750, Ballerup, Denmark

    Morten Grunnet

  4. Department of Medicinal Chemistry, Research Chemist, NeuroSearch A/S, Pederstrupvej 93, DK-2750, Ballerup, Denmark

    Antonio Nardi

Authors
  1. Bo Hjorth Bentzen
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  2. Rune Wederkinck Andersen
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  3. Søren-Peter Olesen
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  4. Morten Grunnet
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  5. Antonio Nardi
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Correspondence to Antonio Nardi.

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Bentzen, B.H., Andersen, R.W., Olesen, SP. et al. Synthesis and characterisation of NS13558: a new important tool for addressing KCa1.1 channel function ex vivo. Naunyn-Schmied Arch Pharmacol 381, 271–283 (2010). https://doi.org/10.1007/s00210-009-0456-2

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