The acrylamide (S)-2 as a positive and negative modulator of Kv7 channels expressed in Xenopus laevis oocytes.
BACKGROUND
Activation of voltage-gated potassium channels of the Kv7 (KCNQ) family reduces cellular excitability. These channels are therefore attractive targets for treatment of diseases characterized by hyperexcitability, such as epilepsy, migraine and neuropathic pain. Retigabine, which opens Kv7.2-5, is now in clinical trial phase III for the treatment of partial onset seizures. One of the main obstacles in developing Kv7 channel active drugs has been to identify compounds that can discriminate between the neuronal subtypes, a feature that could help diminish side effects and increase the potential of drugs for particular indications.


RESULTS
In the present study we have made a thorough investigation of the Bristol-Myers Squibb compound (S)-N-[1-(4-Cyclopropylmethyl-3,4-dihydro-2H-benzo[1], [4]oxazin-6-yl)-ethyl]-3-(2-fluoro-phenyl)-acrylamide [(S)-2] on human Kv7.1-5 channels expressed in Xenopus laevis oocytes. We found that the compound was a weak inhibitor of Kv7.1. In contrast, (S)-2 efficiently opened Kv7.2-5 by producing hyperpolarizing shifts in the voltage-dependence of activation and enhancing the maximal current amplitude. Further, it reduced inactivation, accelerated activation kinetics and slowed deactivation kinetics. The mechanisms of action varied between the subtypes. The enhancing effects of (S)-2 were critically dependent on a tryptophan residue in S5 also known to be crucial for the effects of retigabine, (S)-1 and BMS-204352. However, while (S)-2 did not at all affect a mutant Kv7.4 with a leucine in this position (Kv7.4-W242L), a Kv7.2 with the same mutation (Kv7.2-W236L) was inhibited by the compound, showing that (S)-2 displays a subtype-selective interaction with in the Kv7 family.


CONCLUSIONS
These results offer further insight into pharmacological activation of Kv7 channels, add to the understanding of small molecule interactions with the channels and may contribute to the design of subtype selective modulators.
Blom
Sigrid Marie
SM
Section of Early Target Pharmacology and Physiology, H. Lundbeck A/S, Copenhagen, Denmark.
Schmitt
Nicole
N
Jensen
Henrik Sindal
HS
D020106:Acrylamide / Q000737:chemistry / Q000494:pharmacology*; D000178:Acrylamides / Q000737:chemistry / Q000494:pharmacology*; D000818:Animals; D001665:Binding Sites; D002219:Carbamates / Q000378:metabolism; D006801:Humans; D015640:Ion Channel Gating / Q000187:drug effects; D051656:KCNQ Potassium Channels / Q000378:metabolism*; D007700:Kinetics; D008564:Membrane Potentials / Q000187:drug effects; D050505:Mutant Proteins / Q000378:metabolism; D009865:Oocytes / Q000187:drug effects* / Q000378:metabolism*; D010078:Oxazines / Q000737:chemistry / Q000494:pharmacology*; D010655:Phenylenediamines / Q000378:metabolism; D014364:Tryptophan / Q000378:metabolism; D014982:Xenopus laevis
D000178:Acrylamides; D002219:Carbamates; D051656:KCNQ Potassium Channels; D050505:Mutant Proteins; C492891:N-(1-(4-cyclopropylmethyl-3,4-dihydro-2H-benzo(1,4)oxazin-6-yl)ethyl)-3-(2-fluorophenyl)acrylamide; D010078:Oxazines; D010655:Phenylenediamines; C101866:ezogabine; D020106:Acrylamide; D014364:Tryptophan