Discovery of deoxyceramide analogs as highly selective ACER3 inhibitors in live cells

Acid (AC), neutral (NC) and alkaline ceramidase 3 (ACER3) are the most ubiquitous ceramidases and their therapeutic interest as targets in cancer diseases has been well sustained. This supports the importance of discovering potent and specific inhibitors for further use in combination therapies. Alt...

Descripción completa

Detalles Bibliográficos
Autores: Bielsa, Núria, Casasampere, Mireia, Aseeri, Mazen, Casas, Josefina, Delgado Cirilo, Antonio, Abad, José Luis, Fabriàs, Gemma
Tipo de recurso: artículo
Estado:Versión aceptada para publicación
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:10261/230805
Acceso en línea:http://hdl.handle.net/10261/230805
Access Level:acceso abierto
Palabra clave:Ceramide
Ceramidase
Sphingolipid
Inhibition
Therapeutic target
Enzyme activity
id ES_dc818bd4601e68efe6732dc4195c1c19
oai_identifier_str oai:digital.csic.es:10261/230805
network_acronym_str ES
network_name_str España
repository_id_str
spelling Discovery of deoxyceramide analogs as highly selective ACER3 inhibitors in live cellsBielsa, NúriaCasasampere, MireiaAseeri, MazenCasas, JosefinaDelgado Cirilo, AntonioAbad, José LuisFabriàs, GemmaCeramideCeramidaseSphingolipidInhibitionTherapeutic targetEnzyme activityAcid (AC), neutral (NC) and alkaline ceramidase 3 (ACER3) are the most ubiquitous ceramidases and their therapeutic interest as targets in cancer diseases has been well sustained. This supports the importance of discovering potent and specific inhibitors for further use in combination therapies. Although several ceramidase inhibitors have been reported, most of them target AC and a few focus on NC. In contrast, well characterized ACER3 inhibitors are lacking. Here we report on the synthesis and screening of two series of 1-deoxy(dihydro)ceramide analogs on the three enzymes. Activity was determined using fluorogenic substrates in recombinant human NC (rhNC) and both lysates and intact cells enriched in each enzyme. None of the molecules elicited a remarkable AC inhibitory activity in either experimental setup, while using rhNC, several compounds of both series were active as non-competitive inhibitors with Ki values between 1 and 5 μM. However, a dramatic loss of potency occurred in NC-enriched cell lysates and no activity was elicited in intact cells. Interestingly, several compounds of Series 2 inhibited ACER3 dose-dependently in both cell lysates and intact cells with IC50‘s around 20 μM. In agreement with their activity in live cells, they provoked a significant increase in the amounts of ceramides. Overall, this study identifies highly selective ACER3 activity blockers in intact cells, opening the door to further medicinal chemistry efforts aimed at developing more potent and specific compounds.This work has been partly funded by the Spanish Ministry of Economy, Industry and Competitiveness (grant CTQ2017-85378-R) and Fundación BBVA (grant 35_2018). We thank Dr. Gemma Triola for critically reading the manuscript, and Alexandre Garcia, Pedro Rayo, Neus Roca and Eva Dalmau for their excellent technical assistance.Peer reviewedElsevierMinisterio de Economía y Competitividad (España)Fabriàs, Gemma [0000-0001-7162-3772]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202120212021info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Postprintinfo:eu-repo/semantics/acceptedVersionhttp://hdl.handle.net/10261/230805reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#MICIU/ICTI2017-2020/CTQ2017-85378-Rhttps://doi.org/10.1016/j.ejmech.2021.113296Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2308052026-05-22T06:33:51Z
dc.title.none.fl_str_mv Discovery of deoxyceramide analogs as highly selective ACER3 inhibitors in live cells
title Discovery of deoxyceramide analogs as highly selective ACER3 inhibitors in live cells
spellingShingle Discovery of deoxyceramide analogs as highly selective ACER3 inhibitors in live cells
Bielsa, Núria
Ceramide
Ceramidase
Sphingolipid
Inhibition
Therapeutic target
Enzyme activity
title_short Discovery of deoxyceramide analogs as highly selective ACER3 inhibitors in live cells
title_full Discovery of deoxyceramide analogs as highly selective ACER3 inhibitors in live cells
title_fullStr Discovery of deoxyceramide analogs as highly selective ACER3 inhibitors in live cells
title_full_unstemmed Discovery of deoxyceramide analogs as highly selective ACER3 inhibitors in live cells
title_sort Discovery of deoxyceramide analogs as highly selective ACER3 inhibitors in live cells
dc.creator.none.fl_str_mv Bielsa, Núria
Casasampere, Mireia
Aseeri, Mazen
Casas, Josefina
Delgado Cirilo, Antonio
Abad, José Luis
Fabriàs, Gemma
author Bielsa, Núria
author_facet Bielsa, Núria
Casasampere, Mireia
Aseeri, Mazen
Casas, Josefina
Delgado Cirilo, Antonio
Abad, José Luis
Fabriàs, Gemma
author_role author
author2 Casasampere, Mireia
Aseeri, Mazen
Casas, Josefina
Delgado Cirilo, Antonio
Abad, José Luis
Fabriàs, Gemma
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Economía y Competitividad (España)
Fabriàs, Gemma [0000-0001-7162-3772]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Ceramide
Ceramidase
Sphingolipid
Inhibition
Therapeutic target
Enzyme activity
topic Ceramide
Ceramidase
Sphingolipid
Inhibition
Therapeutic target
Enzyme activity
description Acid (AC), neutral (NC) and alkaline ceramidase 3 (ACER3) are the most ubiquitous ceramidases and their therapeutic interest as targets in cancer diseases has been well sustained. This supports the importance of discovering potent and specific inhibitors for further use in combination therapies. Although several ceramidase inhibitors have been reported, most of them target AC and a few focus on NC. In contrast, well characterized ACER3 inhibitors are lacking. Here we report on the synthesis and screening of two series of 1-deoxy(dihydro)ceramide analogs on the three enzymes. Activity was determined using fluorogenic substrates in recombinant human NC (rhNC) and both lysates and intact cells enriched in each enzyme. None of the molecules elicited a remarkable AC inhibitory activity in either experimental setup, while using rhNC, several compounds of both series were active as non-competitive inhibitors with Ki values between 1 and 5 μM. However, a dramatic loss of potency occurred in NC-enriched cell lysates and no activity was elicited in intact cells. Interestingly, several compounds of Series 2 inhibited ACER3 dose-dependently in both cell lysates and intact cells with IC50‘s around 20 μM. In agreement with their activity in live cells, they provoked a significant increase in the amounts of ceramides. Overall, this study identifies highly selective ACER3 activity blockers in intact cells, opening the door to further medicinal chemistry efforts aimed at developing more potent and specific compounds.
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
Postprint
info:eu-repo/semantics/acceptedVersion
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/230805
url http://hdl.handle.net/10261/230805
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
MICIU/ICTI2017-2020/CTQ2017-85378-R
https://doi.org/10.1016/j.ejmech.2021.113296
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
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
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869421775041855488
score 15,811543

Similares en LA Referencia