Summary
Background and Purpose IC261 (3-[(2,4,6-trimethoxyphenyl)methylidenyl]-indolin-2-one) has previously been introduced as an isoform specific inhibitor of casein kinase 1 (CK1) causing cell cycle arrest or cell death of established tumor cell lines. However, it is reasonable to assume that not all antitumor activities of IC261 are mediated by the inhibition of CK1. Meanwhile there is growing evidence that functional voltage-gated sodium channels are also implicated in the progression of tumors as their blockage suppresses tumor migration and invasion of different tumor cell lines. Thus, we asked whether IC261 functionally inhibits voltage-gated sodium channels. Experimental Approach Electrophysiological experiments were performed using the patch-clamp technique at human heart muscle sodium channels heterologously expressed in human TsA cells. Key Results IC261 inhibits sodium channels in a state-dependent manner. IC261 does not interact with the open channel and has only a low affinity for the resting state of the hNav1.5 (human voltage-gated sodium channel; Kr: 120 μM). The efficacy of IC261 strongly increases with membrane depolarisation, indicating that the inactivated state is an important target. The results of different experimental approaches finally revealed an affinity of IC261 to the inactivated state between 1 and 2 μM. Conclusion and Implications IC261 inhibits sodium channels at a similar concentration necessary to reduce CK1δ/ε activity by 50% (IC50 value 1 μM). Thus, inhibition of sodium channels might contribute to the antitumor activity of IC261.
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Abbreviations
- a(x) :
-
fraction of total
- CK1:
-
casein kinase 1
- CKI-7:
-
N-(2-Aminoethyl)-5-chloroisoquinoline-8-sulphonamide
- D(1–4):
-
domain (1–4)
- [D]:
-
concentration of the drug
- EGTA:
-
ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid
- ENa :
-
equilibrium potential for sodium ions
- g:
-
conductance
- h:
-
relative amount of non-inactivated channels
- HEPES:
-
4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid, N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid)
- hNav(x) :
-
human voltage gated sodium channel (x)
- I:
-
current
- IC :
-
current amplitude under control condition
- IC261:
-
(3-[(2,4,6-trimethoxyphenyl)methylidenyl]-indolin-2-one
- IC50 :
-
concentration of drug which gives half-maximum inhibition
- ID :
-
current amplitude in the presence of drug
- k:
-
slope of the curve
- Kapp :
-
apparent affinity constant
- Kd :
-
drug concentration with half maximal effect
- KI :
-
drug affinity to the inactivated state
- koff :
-
dissociation rate
- kon :
-
association rate
- KR :
-
drug affinity to resting state
- LA:
-
local anesthetic
- MEM:
-
minimum essential medium
- N:
-
Hill coefficient
- N:
-
number of cells
- S(1–4):
-
segment (1–4)
- S:
-
residual current amplitude
- SD:
-
standard deviation
- t:
-
time
- τ:
-
time constant
- TsA201:
-
derivative of the HEK-293 cell line
- TTX:
-
tetrodotoxin
- V:
-
membrane potential (mV)
- V50 :
-
membrane potential of half-maximal activation/inactivation
- VGSC:
-
voltage gated sodium channel
- Ω:
-
ohm, electrical resistance
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Acknowledgement
We thank Ms. Margot Autenrieth-Kronenthaler for her excellent technical assistance.
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Karl Josef Föhr declares that he has no conflict of interest. Uwe Knippschild declares that he has no conflict of interest. Anna Herkommer declares that she has no conflict of interest. Christian Peifer declares that he has no conflict of interest. Michael Fauler declares that he has no conflict of interest. Michael Georgieff declares that he has no conflict of interest. Oliver Adolph declares that he has no conflict of interest.
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The work was supported by the Department of General Surgery and the Department of Anesthesiology, University Hospital of Ulm, Germany.
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This article does not contain any studies with human participants or animals performed by any of the authors.
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Karl J. Föhr and Uwe Knippschild contributed equally to this work.
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Föhr, K.J., Knippschild, U., Herkommer, A. et al. State-dependent block of voltage-gated sodium channels by the casein-kinase 1 inhibitor IC261. Invest New Drugs 35, 277–289 (2017). https://doi.org/10.1007/s10637-017-0429-0
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DOI: https://doi.org/10.1007/s10637-017-0429-0