CB5712809, a novel Keap1 inhibitor upregulates SQSTM1/p62 mediated Nrf2 activation to induce cell death in colon cancer cells

Abstract Background The study focuses on Keap1, a key regulator of the Nrf2 pathway, critical for tumor cell regulation. Recent crystallographic insights into Keap1's structure highlight its functional domains and ligand-binding sites, offering opportunities for targeted drug discovery. The aim...

Full description

Saved in:
Bibliographic Details
Main Authors: Ayed A. Dera, Majed Al Fayi
Format: Article
Language:English
Published: Springer 2025-06-01
Series:Discover Oncology
Subjects:
Online Access:https://doi.org/10.1007/s12672-025-02787-7
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850225851457601536
author Ayed A. Dera
Majed Al Fayi
author_facet Ayed A. Dera
Majed Al Fayi
author_sort Ayed A. Dera
collection DOAJ
description Abstract Background The study focuses on Keap1, a key regulator of the Nrf2 pathway, critical for tumor cell regulation. Recent crystallographic insights into Keap1's structure highlight its functional domains and ligand-binding sites, offering opportunities for targeted drug discovery. The aim is to identify small molecules with high affinity for Keap1 to modulate Keap1, SQSTM1/p62, and Nrf2 functions in colorectal cancer (CRC) cells. Methods A high-throughput virtual screening approach was used to screen the ChemBridge small library against the Keap1 protein. Atomistic Molecular Dynamics (MD) simulations were conducted using GROMACS, along with Gibbs binding free energy estimations. HCT116 and Caco-2 cells were used to determine anti-proliferation. Flow cytometry was used to evaluate target + inhibition in HCT116 and Caco-2 cells. Results The small molecule CB5712809 demonstrated stable interactions with Keap1, supported by molecular dynamics simulations and MM-PBSA analysis. It suppressed the growth of HCT116 and Caco-2 CRC cells with GI50 values of 40.07 nM and 102.80 nM, respectively. Flow cytometry revealed G2/M cell cycle arrest, along with decreased Keap1 levels and increased SQSTM1/p62 and Nrf2 expression, highlighting its potential as a Keap1 modulator. Conclusion Results of this study provide a basis for further experimental validation to develop CB5712809 as a Keap1 targeted chemotherapeutic against CRC. Graphical Abstract
format Article
id doaj-art-9238de7b39ed4ef48c1ddc55ceabefa5
institution OA Journals
issn 2730-6011
language English
publishDate 2025-06-01
publisher Springer
record_format Article
series Discover Oncology
spelling doaj-art-9238de7b39ed4ef48c1ddc55ceabefa52025-08-20T02:05:13ZengSpringerDiscover Oncology2730-60112025-06-0116111510.1007/s12672-025-02787-7CB5712809, a novel Keap1 inhibitor upregulates SQSTM1/p62 mediated Nrf2 activation to induce cell death in colon cancer cellsAyed A. Dera0Majed Al Fayi1Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Khalid UniversityDepartment of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Khalid UniversityAbstract Background The study focuses on Keap1, a key regulator of the Nrf2 pathway, critical for tumor cell regulation. Recent crystallographic insights into Keap1's structure highlight its functional domains and ligand-binding sites, offering opportunities for targeted drug discovery. The aim is to identify small molecules with high affinity for Keap1 to modulate Keap1, SQSTM1/p62, and Nrf2 functions in colorectal cancer (CRC) cells. Methods A high-throughput virtual screening approach was used to screen the ChemBridge small library against the Keap1 protein. Atomistic Molecular Dynamics (MD) simulations were conducted using GROMACS, along with Gibbs binding free energy estimations. HCT116 and Caco-2 cells were used to determine anti-proliferation. Flow cytometry was used to evaluate target + inhibition in HCT116 and Caco-2 cells. Results The small molecule CB5712809 demonstrated stable interactions with Keap1, supported by molecular dynamics simulations and MM-PBSA analysis. It suppressed the growth of HCT116 and Caco-2 CRC cells with GI50 values of 40.07 nM and 102.80 nM, respectively. Flow cytometry revealed G2/M cell cycle arrest, along with decreased Keap1 levels and increased SQSTM1/p62 and Nrf2 expression, highlighting its potential as a Keap1 modulator. Conclusion Results of this study provide a basis for further experimental validation to develop CB5712809 as a Keap1 targeted chemotherapeutic against CRC. Graphical Abstracthttps://doi.org/10.1007/s12672-025-02787-7Keap1Nrf2 pathwayVirtual screeningMolecular dynamics simulationProtein–ligand interactionCancer therapy
spellingShingle Ayed A. Dera
Majed Al Fayi
CB5712809, a novel Keap1 inhibitor upregulates SQSTM1/p62 mediated Nrf2 activation to induce cell death in colon cancer cells
Discover Oncology
Keap1
Nrf2 pathway
Virtual screening
Molecular dynamics simulation
Protein–ligand interaction
Cancer therapy
title CB5712809, a novel Keap1 inhibitor upregulates SQSTM1/p62 mediated Nrf2 activation to induce cell death in colon cancer cells
title_full CB5712809, a novel Keap1 inhibitor upregulates SQSTM1/p62 mediated Nrf2 activation to induce cell death in colon cancer cells
title_fullStr CB5712809, a novel Keap1 inhibitor upregulates SQSTM1/p62 mediated Nrf2 activation to induce cell death in colon cancer cells
title_full_unstemmed CB5712809, a novel Keap1 inhibitor upregulates SQSTM1/p62 mediated Nrf2 activation to induce cell death in colon cancer cells
title_short CB5712809, a novel Keap1 inhibitor upregulates SQSTM1/p62 mediated Nrf2 activation to induce cell death in colon cancer cells
title_sort cb5712809 a novel keap1 inhibitor upregulates sqstm1 p62 mediated nrf2 activation to induce cell death in colon cancer cells
topic Keap1
Nrf2 pathway
Virtual screening
Molecular dynamics simulation
Protein–ligand interaction
Cancer therapy
url https://doi.org/10.1007/s12672-025-02787-7
work_keys_str_mv AT ayedadera cb5712809anovelkeap1inhibitorupregulatessqstm1p62mediatednrf2activationtoinducecelldeathincoloncancercells
AT majedalfayi cb5712809anovelkeap1inhibitorupregulatessqstm1p62mediatednrf2activationtoinducecelldeathincoloncancercells