Phenylalanine-Derived β-Lactam TRPM8 Modulators. Configuration Effect on the Antagonist Activity

Transient receptor potential cation channel subfamily M member 8 (TRPM8) is a Ca2+ nonselective ion channel implicated in a variety of pathological conditions, including cancer, inflammatory and neuropathic pain. In previous works we identified a family of chiral, highly hydrophobic β–lactam derivat...

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Autores: Bonache de Marcos, María Ángeles, Llabrés, Pedro Juan, Martín-Escura, Cristina, Torre-Martínez, Roberto de la, Medina-Peris, Alicia, Butrón, Laura, Gómez-Monterrey, Isabel, Roa, Ana María, Fernández-Ballester, Gregorio, Ferrer-Montiel, Antonio, Fernández-Carvajal, Asia, González-Muñiz, Rosario
Tipo de recurso: artículo
Fecha de publicación:2021
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/240941
Acceso en línea:http://hdl.handle.net/10261/240941
Access Level:acceso abierto
Palabra clave:TRPM8
Antagonists
β–lactams
Absolute configuration
Ca2+ microfluorimetry
Patch- Clamp
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spelling Phenylalanine-Derived β-Lactam TRPM8 Modulators. Configuration Effect on the Antagonist ActivityBonache de Marcos, María ÁngelesLlabrés, Pedro JuanMartín-Escura, CristinaTorre-Martínez, Roberto de laMedina-Peris, AliciaButrón, LauraGómez-Monterrey, IsabelRoa, Ana MaríaFernández-Ballester, GregorioFerrer-Montiel, AntonioFernández-Carvajal, AsiaGonzález-Muñiz, RosarioTRPM8Antagonistsβ–lactamsAbsolute configurationCa2+ microfluorimetryPatch- ClampTransient receptor potential cation channel subfamily M member 8 (TRPM8) is a Ca2+ nonselective ion channel implicated in a variety of pathological conditions, including cancer, inflammatory and neuropathic pain. In previous works we identified a family of chiral, highly hydrophobic β–lactam derivatives, and began to intuit a possible effect of the stereogenic centers on the antagonist activity. To investigate the influence of configuration on the TRPM8 antagonist properties, here we prepare and characterize four possible diastereoisomeric derivatives of 4-benzyl-1-[(3’-phenyl- 2’-dibenzylamino)prop-1’-yl]-4-benzyloxycarbonyl-3-methyl-2-oxoazetidine. In microfluorography assays, all isomers were able to reduce the menthol-induced cell Ca2+ entry to larger or lesser extent. Potency follows the order 3R,4R,2’R > 3S,4S,2’R ≅ 3R,4R,2’S > 3S,4S,2’S, with the most potent diastereoisomer showing a half inhibitory concentration (IC50) in the low nanomolar range, confirmed by Patch-Clamp electrophysiology experiments. All four compounds display high receptor selectivity against other members of the TRP family. Furthermore, in primary cultures of rat dorsal root ganglion (DRG) neurons, the most potent diastereoisomers do not produce any alteration in neuronal excitability, indicating their high specificity for TRPM8 channels. Docking studies positioned these β-lactams at different subsites by the pore zone, suggesting a different mechanism than the known N-(3-aminopropyl)-2-[(3-methylphenyl)methoxy]-N-(2-thienylmethyl)-benzamide (AMTB) antagonist.This research was funded by the Spanish Ministerio de Ciencia y Universidades (MICYU-FEDER, RTI2018-097189-C2-1 to A.F.-M. and A.F.-C., and RTI2018-097189-C2-2 to R.G.M.), Comunidad de Madrid (IND2017/BMD7673 to R.G.M.) and the Spanish National Research Council (CSIC, 201980E030 to R.G.M.).Peer reviewedMultidisciplinary Digital Publishing InstituteComunidad de MadridEuropean CommissionMinisterio de Ciencia, Innovación y Universidades (España)Bonache, María Ángeles [0000-0002-4922-5345]González-Muñiz, Rosario [0000-0001-8833-4328]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202120212021info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501http://hdl.handle.net/10261/240941reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-097189-C2-1info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-097189-C2-2https://doi.org/10.3390/ijms22052370Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2409412026-05-22T06:33:51Z
dc.title.none.fl_str_mv Phenylalanine-Derived β-Lactam TRPM8 Modulators. Configuration Effect on the Antagonist Activity
title Phenylalanine-Derived β-Lactam TRPM8 Modulators. Configuration Effect on the Antagonist Activity
spellingShingle Phenylalanine-Derived β-Lactam TRPM8 Modulators. Configuration Effect on the Antagonist Activity
Bonache de Marcos, María Ángeles
TRPM8
Antagonists
β–lactams
Absolute configuration
Ca2+ microfluorimetry
Patch- Clamp
title_short Phenylalanine-Derived β-Lactam TRPM8 Modulators. Configuration Effect on the Antagonist Activity
title_full Phenylalanine-Derived β-Lactam TRPM8 Modulators. Configuration Effect on the Antagonist Activity
title_fullStr Phenylalanine-Derived β-Lactam TRPM8 Modulators. Configuration Effect on the Antagonist Activity
title_full_unstemmed Phenylalanine-Derived β-Lactam TRPM8 Modulators. Configuration Effect on the Antagonist Activity
title_sort Phenylalanine-Derived β-Lactam TRPM8 Modulators. Configuration Effect on the Antagonist Activity
dc.creator.none.fl_str_mv Bonache de Marcos, María Ángeles
Llabrés, Pedro Juan
Martín-Escura, Cristina
Torre-Martínez, Roberto de la
Medina-Peris, Alicia
Butrón, Laura
Gómez-Monterrey, Isabel
Roa, Ana María
Fernández-Ballester, Gregorio
Ferrer-Montiel, Antonio
Fernández-Carvajal, Asia
González-Muñiz, Rosario
author Bonache de Marcos, María Ángeles
author_facet Bonache de Marcos, María Ángeles
Llabrés, Pedro Juan
Martín-Escura, Cristina
Torre-Martínez, Roberto de la
Medina-Peris, Alicia
Butrón, Laura
Gómez-Monterrey, Isabel
Roa, Ana María
Fernández-Ballester, Gregorio
Ferrer-Montiel, Antonio
Fernández-Carvajal, Asia
González-Muñiz, Rosario
author_role author
author2 Llabrés, Pedro Juan
Martín-Escura, Cristina
Torre-Martínez, Roberto de la
Medina-Peris, Alicia
Butrón, Laura
Gómez-Monterrey, Isabel
Roa, Ana María
Fernández-Ballester, Gregorio
Ferrer-Montiel, Antonio
Fernández-Carvajal, Asia
González-Muñiz, Rosario
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Comunidad de Madrid
European Commission
Ministerio de Ciencia, Innovación y Universidades (España)
Bonache, María Ángeles [0000-0002-4922-5345]
González-Muñiz, Rosario [0000-0001-8833-4328]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv TRPM8
Antagonists
β–lactams
Absolute configuration
Ca2+ microfluorimetry
Patch- Clamp
topic TRPM8
Antagonists
β–lactams
Absolute configuration
Ca2+ microfluorimetry
Patch- Clamp
description Transient receptor potential cation channel subfamily M member 8 (TRPM8) is a Ca2+ nonselective ion channel implicated in a variety of pathological conditions, including cancer, inflammatory and neuropathic pain. In previous works we identified a family of chiral, highly hydrophobic β–lactam derivatives, and began to intuit a possible effect of the stereogenic centers on the antagonist activity. To investigate the influence of configuration on the TRPM8 antagonist properties, here we prepare and characterize four possible diastereoisomeric derivatives of 4-benzyl-1-[(3’-phenyl- 2’-dibenzylamino)prop-1’-yl]-4-benzyloxycarbonyl-3-methyl-2-oxoazetidine. In microfluorography assays, all isomers were able to reduce the menthol-induced cell Ca2+ entry to larger or lesser extent. Potency follows the order 3R,4R,2’R > 3S,4S,2’R ≅ 3R,4R,2’S > 3S,4S,2’S, with the most potent diastereoisomer showing a half inhibitory concentration (IC50) in the low nanomolar range, confirmed by Patch-Clamp electrophysiology experiments. All four compounds display high receptor selectivity against other members of the TRP family. Furthermore, in primary cultures of rat dorsal root ganglion (DRG) neurons, the most potent diastereoisomers do not produce any alteration in neuronal excitability, indicating their high specificity for TRPM8 channels. Docking studies positioned these β-lactams at different subsites by the pore zone, suggesting a different mechanism than the known N-(3-aminopropyl)-2-[(3-methylphenyl)methoxy]-N-(2-thienylmethyl)-benzamide (AMTB) antagonist.
publishDate 2021
dc.date.none.fl_str_mv 2021
2021
2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/240941
url http://hdl.handle.net/10261/240941
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-097189-C2-1
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-097189-C2-2
https://doi.org/10.3390/ijms22052370

dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
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