Cargando…

MEPIRAPIM-derived synthetic cannabinoids inhibit T-type calcium channels with divergent effects on seizures in rodent models of epilepsy

Background: T-type Ca(2+) channels (Ca(v)3) represent emerging therapeutic targets for a range of neurological disorders, including epilepsy and pain. To aid the development and optimisation of new therapeutics, there is a need to identify novel chemical entities which act at these ion channels. A n...

Descripción completa

Detalles Bibliográficos
Autores principales: Harman, Thomas, Udoh, Michael, McElroy, Dan L., Anderson, Lyndsey L., Kevin, Richard C., Banister, Samuel D., Ametovski, Adam, Markham, Jack, Bladen, Chris, Doohan, Peter T., Greba, Quentin, Laprairie, Robert B., Snutch, Terrance P., McGregor, Iain S., Howland, John G., Arnold, Jonathon C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10110893/
https://www.ncbi.nlm.nih.gov/pubmed/37082241
http://dx.doi.org/10.3389/fphys.2023.1086243
_version_ 1785027340649103360
author Harman, Thomas
Udoh, Michael
McElroy, Dan L.
Anderson, Lyndsey L.
Kevin, Richard C.
Banister, Samuel D.
Ametovski, Adam
Markham, Jack
Bladen, Chris
Doohan, Peter T.
Greba, Quentin
Laprairie, Robert B.
Snutch, Terrance P.
McGregor, Iain S.
Howland, John G.
Arnold, Jonathon C.
author_facet Harman, Thomas
Udoh, Michael
McElroy, Dan L.
Anderson, Lyndsey L.
Kevin, Richard C.
Banister, Samuel D.
Ametovski, Adam
Markham, Jack
Bladen, Chris
Doohan, Peter T.
Greba, Quentin
Laprairie, Robert B.
Snutch, Terrance P.
McGregor, Iain S.
Howland, John G.
Arnold, Jonathon C.
author_sort Harman, Thomas
collection PubMed
description Background: T-type Ca(2+) channels (Ca(v)3) represent emerging therapeutic targets for a range of neurological disorders, including epilepsy and pain. To aid the development and optimisation of new therapeutics, there is a need to identify novel chemical entities which act at these ion channels. A number of synthetic cannabinoid receptor agonists (SCRAs) have been found to exhibit activity at T-type channels, suggesting that cannabinoids may provide convenient chemical scaffolds on which to design novel Ca(v)3 inhibitors. However, activity at cannabinoid type 1 (CB(1)) receptors can be problematic because of central and peripheral toxicities associated with potent SCRAs. The putative SCRA MEPIRAPIM and its analogues were recently identified as Ca(v)3 inhibitors with only minimal activity at CB(1) receptors, opening the possibility that this scaffold may be exploited to develop novel, selective Ca(v)3 inhibitors. Here we present the pharmacological characterisation of SB2193 and SB2193F, two novel Ca(v)3 inhibitors derived from MEPIRAPIM. Methods: The potency of SB2193 and SB2193F was evaluated in vitro using a fluorometric Ca(2+) flux assay and confirmed using whole-cell patch-clamp electrophysiology. In silico docking to the cryo-EM structure of Ca(v)3.1 was also performed to elucidate structural insights into T-type channel inhibition. Next, in vivo pharmacokinetic parameters in mouse brain and plasma were determined using liquid chromatography-mass spectroscopy. Finally, anticonvulsant activity was assayed in established genetic and electrically-induced rodent seizure models. Results: Both MEPIRAPIM derivatives produced potent inhibition of Ca(v)3 channels and were brain penetrant, with SB2193 exhibiting a brain/plasma ratio of 2.7. SB2193 was further examined in mouse seizure models where it acutely protected against 6 Hz-induced seizures. However, SB2193 did not reduce spontaneous seizures in the Scn1a ( +/− ) mouse model of Dravet syndrome, nor absence seizures in the Genetic Absence Epilepsy Rat from Strasbourg (GAERS). Surprisingly, SB2193 appeared to increase the incidence and duration of spike-and-wave discharges in GAERS animals over a 4 h recording period. Conclusion: These results show that MEPIRAPIM analogues provide novel chemical scaffolds to advance Ca(v)3 inhibitors against certain seizure types.
format Online
Article
Text
id pubmed-10110893
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-101108932023-04-19 MEPIRAPIM-derived synthetic cannabinoids inhibit T-type calcium channels with divergent effects on seizures in rodent models of epilepsy Harman, Thomas Udoh, Michael McElroy, Dan L. Anderson, Lyndsey L. Kevin, Richard C. Banister, Samuel D. Ametovski, Adam Markham, Jack Bladen, Chris Doohan, Peter T. Greba, Quentin Laprairie, Robert B. Snutch, Terrance P. McGregor, Iain S. Howland, John G. Arnold, Jonathon C. Front Physiol Physiology Background: T-type Ca(2+) channels (Ca(v)3) represent emerging therapeutic targets for a range of neurological disorders, including epilepsy and pain. To aid the development and optimisation of new therapeutics, there is a need to identify novel chemical entities which act at these ion channels. A number of synthetic cannabinoid receptor agonists (SCRAs) have been found to exhibit activity at T-type channels, suggesting that cannabinoids may provide convenient chemical scaffolds on which to design novel Ca(v)3 inhibitors. However, activity at cannabinoid type 1 (CB(1)) receptors can be problematic because of central and peripheral toxicities associated with potent SCRAs. The putative SCRA MEPIRAPIM and its analogues were recently identified as Ca(v)3 inhibitors with only minimal activity at CB(1) receptors, opening the possibility that this scaffold may be exploited to develop novel, selective Ca(v)3 inhibitors. Here we present the pharmacological characterisation of SB2193 and SB2193F, two novel Ca(v)3 inhibitors derived from MEPIRAPIM. Methods: The potency of SB2193 and SB2193F was evaluated in vitro using a fluorometric Ca(2+) flux assay and confirmed using whole-cell patch-clamp electrophysiology. In silico docking to the cryo-EM structure of Ca(v)3.1 was also performed to elucidate structural insights into T-type channel inhibition. Next, in vivo pharmacokinetic parameters in mouse brain and plasma were determined using liquid chromatography-mass spectroscopy. Finally, anticonvulsant activity was assayed in established genetic and electrically-induced rodent seizure models. Results: Both MEPIRAPIM derivatives produced potent inhibition of Ca(v)3 channels and were brain penetrant, with SB2193 exhibiting a brain/plasma ratio of 2.7. SB2193 was further examined in mouse seizure models where it acutely protected against 6 Hz-induced seizures. However, SB2193 did not reduce spontaneous seizures in the Scn1a ( +/− ) mouse model of Dravet syndrome, nor absence seizures in the Genetic Absence Epilepsy Rat from Strasbourg (GAERS). Surprisingly, SB2193 appeared to increase the incidence and duration of spike-and-wave discharges in GAERS animals over a 4 h recording period. Conclusion: These results show that MEPIRAPIM analogues provide novel chemical scaffolds to advance Ca(v)3 inhibitors against certain seizure types. Frontiers Media S.A. 2023-04-04 /pmc/articles/PMC10110893/ /pubmed/37082241 http://dx.doi.org/10.3389/fphys.2023.1086243 Text en Copyright © 2023 Harman, Udoh, McElroy, Anderson, Kevin, Banister, Ametovski, Markham, Bladen, Doohan, Greba, Laprairie, Snutch, McGregor, Howland and Arnold. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Physiology
Harman, Thomas
Udoh, Michael
McElroy, Dan L.
Anderson, Lyndsey L.
Kevin, Richard C.
Banister, Samuel D.
Ametovski, Adam
Markham, Jack
Bladen, Chris
Doohan, Peter T.
Greba, Quentin
Laprairie, Robert B.
Snutch, Terrance P.
McGregor, Iain S.
Howland, John G.
Arnold, Jonathon C.
MEPIRAPIM-derived synthetic cannabinoids inhibit T-type calcium channels with divergent effects on seizures in rodent models of epilepsy
title MEPIRAPIM-derived synthetic cannabinoids inhibit T-type calcium channels with divergent effects on seizures in rodent models of epilepsy
title_full MEPIRAPIM-derived synthetic cannabinoids inhibit T-type calcium channels with divergent effects on seizures in rodent models of epilepsy
title_fullStr MEPIRAPIM-derived synthetic cannabinoids inhibit T-type calcium channels with divergent effects on seizures in rodent models of epilepsy
title_full_unstemmed MEPIRAPIM-derived synthetic cannabinoids inhibit T-type calcium channels with divergent effects on seizures in rodent models of epilepsy
title_short MEPIRAPIM-derived synthetic cannabinoids inhibit T-type calcium channels with divergent effects on seizures in rodent models of epilepsy
title_sort mepirapim-derived synthetic cannabinoids inhibit t-type calcium channels with divergent effects on seizures in rodent models of epilepsy
topic Physiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10110893/
https://www.ncbi.nlm.nih.gov/pubmed/37082241
http://dx.doi.org/10.3389/fphys.2023.1086243
work_keys_str_mv AT harmanthomas mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy
AT udohmichael mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy
AT mcelroydanl mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy
AT andersonlyndseyl mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy
AT kevinrichardc mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy
AT banistersamueld mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy
AT ametovskiadam mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy
AT markhamjack mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy
AT bladenchris mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy
AT doohanpetert mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy
AT grebaquentin mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy
AT laprairierobertb mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy
AT snutchterrancep mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy
AT mcgregoriains mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy
AT howlandjohng mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy
AT arnoldjonathonc mepirapimderivedsyntheticcannabinoidsinhibitttypecalciumchannelswithdivergenteffectsonseizuresinrodentmodelsofepilepsy