A novel β-TrCP1/NRF2 interaction inhibitor for effective anti-inflammatory therapy

Background Non-communicable chronic diseases are characterized by low-grade inflammation and oxidative stress. Extensive research has identified the transcription factor NRF2 as a potential therapeutic target. Current NRF2 activators, designed to inhibit its repressor KEAP1, often exhibit undesirabl...

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Autores: García Yagüe, Ángel Juan, Cañizares Moscato, Lucía, Encinar, José Antonio, Cazalla Ibáñez, Eduardo, Fernández Ginés, Raquel, Escoll, Maribel, Rojo Sanchís, Ana Isabel, Cuadrado Pastor, Antonio
Tipo de recurso: artículo
Fecha de publicación:2025
País:España
Institución:Universidad Autónoma de Madrid
Repositorio:Biblos-e Archivo. Repositorio Institucional de la UAM
Idioma:inglés
OAI Identifier:oai:repositorio.uam.es:10486/728500
Acceso en línea:https://hdl.handle.net/10486/728500
https://dx.doi.org/10.1186/s12929-025-01157-3
Access Level:acceso abierto
Palabra clave:NRF2
β-TrCP1
Protein–protein interaction-inhibitor
Inflammation
Liver
Medicina
Química
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oai_identifier_str oai:repositorio.uam.es:10486/728500
network_acronym_str ES
network_name_str España
repository_id_str
dc.title.none.fl_str_mv A novel β-TrCP1/NRF2 interaction inhibitor for effective anti-inflammatory therapy
title A novel β-TrCP1/NRF2 interaction inhibitor for effective anti-inflammatory therapy
spellingShingle A novel β-TrCP1/NRF2 interaction inhibitor for effective anti-inflammatory therapy
García Yagüe, Ángel Juan
NRF2
β-TrCP1
Protein–protein interaction-inhibitor
Inflammation
Liver
Medicina
Química
title_short A novel β-TrCP1/NRF2 interaction inhibitor for effective anti-inflammatory therapy
title_full A novel β-TrCP1/NRF2 interaction inhibitor for effective anti-inflammatory therapy
title_fullStr A novel β-TrCP1/NRF2 interaction inhibitor for effective anti-inflammatory therapy
title_full_unstemmed A novel β-TrCP1/NRF2 interaction inhibitor for effective anti-inflammatory therapy
title_sort A novel β-TrCP1/NRF2 interaction inhibitor for effective anti-inflammatory therapy
dc.creator.none.fl_str_mv García Yagüe, Ángel Juan
Cañizares Moscato, Lucía
Encinar, José Antonio
Cazalla Ibáñez, Eduardo
Fernández Ginés, Raquel
Escoll, Maribel
Rojo Sanchís, Ana Isabel
Cuadrado Pastor, Antonio
author García Yagüe, Ángel Juan
author_facet García Yagüe, Ángel Juan
Cañizares Moscato, Lucía
Encinar, José Antonio
Cazalla Ibáñez, Eduardo
Fernández Ginés, Raquel
Escoll, Maribel
Rojo Sanchís, Ana Isabel
Cuadrado Pastor, Antonio
author_role author
author2 Cañizares Moscato, Lucía
Encinar, José Antonio
Cazalla Ibáñez, Eduardo
Fernández Ginés, Raquel
Escoll, Maribel
Rojo Sanchís, Ana Isabel
Cuadrado Pastor, Antonio
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Departamento de Bioquímica
Facultad de Medicina
Gobierno de España
Comunidad de Madrid
dc.subject.none.fl_str_mv NRF2
β-TrCP1
Protein–protein interaction-inhibitor
Inflammation
Liver
Medicina
Química
topic NRF2
β-TrCP1
Protein–protein interaction-inhibitor
Inflammation
Liver
Medicina
Química
description Background Non-communicable chronic diseases are characterized by low-grade inflammation and oxidative stress. Extensive research has identified the transcription factor NRF2 as a potential therapeutic target. Current NRF2 activators, designed to inhibit its repressor KEAP1, often exhibit undesirable side effects. As an alternative approach, we previously developed PHAR, a protein–protein interaction inhibitor of β-TrCP1/NRF2, which promotes NRF2 activation. Using the same in silico screening platform, we have now identified a novel compound, P10. This small molecule selectively interferes with the β-TrCP1/NRF2 interaction, leading to NRF2 stabilization and transcriptional activation of its target genes in a β-TrCP1-dependent manner, demonstrating promising effects in a liver model of acute inflammation. Methods After an in silico screening of ∼1 million compounds, including molecular docking analysis, ADMET evaluation, and molecular dynamics simulations, we identified and characterized a novel small molecule, P10, which inhibits β-TrCP1/NRF2 interaction. The compound was validated using luciferase reporter assays, co-immunoprecipitation, and ubiquitination experiments. The specificity of P10 was assessed by comparing NRF2 signatures in wild-type and Nrf2-null cells. The impact of NRF2 activation induced by P10 was investigated by evaluating its antioxidant and anti-inflammatory responses against tert-butyl hydroperoxide and lipopolysaccharide, respectively. Finally, wild-type and Nrf2-null mice were administered P10 intraperitoneally at a dose of 20 mg/kg daily for five consecutive days. Four hours before sacrifice, all animals received a lipopolysaccharide (LPS) injection at 10 mg/kg. Results P10 selectively disrupts the interaction between β-TrCP1 and NRF2, thereby inhibiting β-TrCP1-mediated ubiquitination of NRF2 and leading to the upregulation of NRF2 target genes. Additionally, P10 mitigates oxidative stress induced by tert-butyl hydroperoxide and reduces pro-inflammatory markers in an NRF2-dependent manner in macrophages treated with lipopolysaccharide. In a preclinical model of liver inflammation, P10 specifically targets the liver, significantly attenuating lipopolysaccharide-induced inflammation through the activation of NRF2. This is demonstrated by decreased expression of inflammatory cytokine genes and a reduction in F4/80-stained liver macrophages. Notably, this anti-inflammatory effect is absent in Nrf2-knockout mice, confirming its NRF2-dependent mechanism of action. Conclusions P10 emerges as a promising NRF2 activator by selectively disrupting the β-TrCP1/NRF2 interaction, highlighting its potential as a therapeutic agent for diseases presenting acute liver inflammation
publishDate 2025
dc.date.none.fl_str_mv 2025
2025-07-11
dc.type.none.fl_str_mv research article
http://purl.org/coar/resource_type/c_2df8fbb1
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/10486/728500
https://dx.doi.org/10.1186/s12929-025-01157-3
40646550
url https://hdl.handle.net/10486/728500
https://dx.doi.org/10.1186/s12929-025-01157-3
identifier_str_mv 40646550
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Springer Nature
publisher.none.fl_str_mv Springer Nature
dc.source.none.fl_str_mv reponame:Biblos-e Archivo. Repositorio Institucional de la UAM
instname:Universidad Autónoma de Madrid
instname_str Universidad Autónoma de Madrid
reponame_str Biblos-e Archivo. Repositorio Institucional de la UAM
collection Biblos-e Archivo. Repositorio Institucional de la UAM
repository.name.fl_str_mv
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spelling A novel β-TrCP1/NRF2 interaction inhibitor for effective anti-inflammatory therapyGarcía Yagüe, Ángel JuanCañizares Moscato, LucíaEncinar, José AntonioCazalla Ibáñez, EduardoFernández Ginés, RaquelEscoll, MaribelRojo Sanchís, Ana IsabelCuadrado Pastor, AntonioNRF2β-TrCP1Protein–protein interaction-inhibitorInflammationLiverMedicinaQuímicaBackground Non-communicable chronic diseases are characterized by low-grade inflammation and oxidative stress. Extensive research has identified the transcription factor NRF2 as a potential therapeutic target. Current NRF2 activators, designed to inhibit its repressor KEAP1, often exhibit undesirable side effects. As an alternative approach, we previously developed PHAR, a protein–protein interaction inhibitor of β-TrCP1/NRF2, which promotes NRF2 activation. Using the same in silico screening platform, we have now identified a novel compound, P10. This small molecule selectively interferes with the β-TrCP1/NRF2 interaction, leading to NRF2 stabilization and transcriptional activation of its target genes in a β-TrCP1-dependent manner, demonstrating promising effects in a liver model of acute inflammation. Methods After an in silico screening of ∼1 million compounds, including molecular docking analysis, ADMET evaluation, and molecular dynamics simulations, we identified and characterized a novel small molecule, P10, which inhibits β-TrCP1/NRF2 interaction. The compound was validated using luciferase reporter assays, co-immunoprecipitation, and ubiquitination experiments. The specificity of P10 was assessed by comparing NRF2 signatures in wild-type and Nrf2-null cells. The impact of NRF2 activation induced by P10 was investigated by evaluating its antioxidant and anti-inflammatory responses against tert-butyl hydroperoxide and lipopolysaccharide, respectively. Finally, wild-type and Nrf2-null mice were administered P10 intraperitoneally at a dose of 20 mg/kg daily for five consecutive days. Four hours before sacrifice, all animals received a lipopolysaccharide (LPS) injection at 10 mg/kg. Results P10 selectively disrupts the interaction between β-TrCP1 and NRF2, thereby inhibiting β-TrCP1-mediated ubiquitination of NRF2 and leading to the upregulation of NRF2 target genes. Additionally, P10 mitigates oxidative stress induced by tert-butyl hydroperoxide and reduces pro-inflammatory markers in an NRF2-dependent manner in macrophages treated with lipopolysaccharide. In a preclinical model of liver inflammation, P10 specifically targets the liver, significantly attenuating lipopolysaccharide-induced inflammation through the activation of NRF2. This is demonstrated by decreased expression of inflammatory cytokine genes and a reduction in F4/80-stained liver macrophages. Notably, this anti-inflammatory effect is absent in Nrf2-knockout mice, confirming its NRF2-dependent mechanism of action. Conclusions P10 emerges as a promising NRF2 activator by selectively disrupting the β-TrCP1/NRF2 interaction, highlighting its potential as a therapeutic agent for diseases presenting acute liver inflammationThis research was funded by the Spanish Ministry of Economy and Competitiveness (MINECO) (grants PDC2021-121421-I00, PDC2022-1337665-I00, PID2022-141786OB-I00, and PID2019-110061RB-I00). The Autonomous Community of Madrid (grants S2017BMD-3827 and P2022_BMD-7230). EC is the holder of an FPU contract of MIU (Ministry of Universities FPU2021, FPU21/02505)Springer NatureDepartamento de BioquímicaFacultad de MedicinaGobierno de EspañaComunidad de Madrid20252025-07-11research articlehttp://purl.org/coar/resource_type/c_2df8fbb1VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/10486/728500https://dx.doi.org/10.1186/s12929-025-01157-340646550reponame:Biblos-e Archivo. Repositorio Institucional de la UAMinstname:Universidad Autónoma de MadridInglésengopen accesshttp://purl.org/coar/access_right/c_abf2Attribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:repositorio.uam.es:10486/7285002026-06-23T12:46:27Z
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