In Silico and in vitro evaluation of the anticancer effect of a 1,5-Benzodiazepin-2-One derivative (3b) revealing potent dual inhibition of HER2 and HDAC1

Abstract Benzodiazepines are widely recognized for their therapeutic benefits in the treatment of anxiety and insomnia. However, in the pursuit of innovative anticancer agents, they have gained attention as a possible pharmacophore. One of those promising anticancer benzodiazepines is 3b which was d...

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Main Authors: Nada S. Al-Theyab, Ahmad H. Bakheit, Sanae Lahmidi, Mohammed Mufadhe Alanazi, Awadh M Ali, Khalil Azzaoui, El Mokhtar Essassi, Joel T. Mague, Mohamad Hefnawy, Mohammed Mater Alanazi, Ashwag S. Alanazi
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-97786-7
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author Nada S. Al-Theyab
Ahmad H. Bakheit
Sanae Lahmidi
Mohammed Mufadhe Alanazi
Awadh M Ali
Khalil Azzaoui
El Mokhtar Essassi
Joel T. Mague
Mohamad Hefnawy
Mohammed Mater Alanazi
Ashwag S. Alanazi
author_facet Nada S. Al-Theyab
Ahmad H. Bakheit
Sanae Lahmidi
Mohammed Mufadhe Alanazi
Awadh M Ali
Khalil Azzaoui
El Mokhtar Essassi
Joel T. Mague
Mohamad Hefnawy
Mohammed Mater Alanazi
Ashwag S. Alanazi
author_sort Nada S. Al-Theyab
collection DOAJ
description Abstract Benzodiazepines are widely recognized for their therapeutic benefits in the treatment of anxiety and insomnia. However, in the pursuit of innovative anticancer agents, they have gained attention as a possible pharmacophore. One of those promising anticancer benzodiazepines is 3b which was demonstrated to exert good antiproliferative effects. To investigate the anticancer effect of 3b, in silico prediction of the possible targets were performed. Then, the predicted targets were investigated through in vitro study. Furthermore, 3b was evaluated for its effects on cell cycle suppression and induction of apoptosis. Molecular docking was used to study the possible types of interactions while molecular dynamics simulations were conducted to estimate the protein–ligand complex’s stability and dynamic behavior. Results demonstrated that 3b is a potent dual inhibitor of HER2 and HDAC1 with IC50 values of 0.023 and 0.041 nM, respectively. Moreover, 3b was found to suppress cell cycle progression in G2/M phase and induce early and late apoptosis in HepG2 cancer cells. Further analysis of apoptotic markers revealed an induction of Caspase 3 and BAX proapoptotic proteins along with a suppression of the antiapoptotic protein (Bcl-2). Molecular docking of 3b into the active site of HER2 and HDAC1 displayed significant types of interactions with active sites of these target proteins while molecular dynamics simulations demonstrated the overall structural stability of HER2 and HDAC1 is maintained or even enhanced upon ligand binding. In conclusion, 3b is a powerful anticancer agent that exerts its effects by inhibiting HDAC1 and HER2, resulting in cell cycle arrest and cancer cell death through apoptosis. Nonetheless, additional investigations are needed to explore its mechanisms and therapeutic efficacy in more detail.
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spelling doaj-art-5e0879a2e62b4cd9a21e8c670fb2ff9d2025-08-20T03:18:27ZengNature PortfolioScientific Reports2045-23222025-04-0115111410.1038/s41598-025-97786-7In Silico and in vitro evaluation of the anticancer effect of a 1,5-Benzodiazepin-2-One derivative (3b) revealing potent dual inhibition of HER2 and HDAC1Nada S. Al-Theyab0Ahmad H. Bakheit1Sanae Lahmidi2Mohammed Mufadhe Alanazi3Awadh M Ali4Khalil Azzaoui5El Mokhtar Essassi6Joel T. Mague7Mohamad Hefnawy8Mohammed Mater Alanazi9Ashwag S. Alanazi10Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud UniversityDepartment of Pharmaceutical Chemistry, College of Pharmacy, King Saud UniversityLaboratory of Heterocyclic Organic Chemistry, Department of Chemistry, Faculty of Sciences, Mohammed V University in RabatDepartment of Pharmacology and Toxicology, College of Pharmacy, King Saud UniversityDepartment of Pharmaceutical Chemistry, College of Pharmacy, King Saud UniversityEngineering Laboratory of Organometallic, Molecular Materials, and Environment, Faculty of Sciences, University Sidi Mohamed Ben AbdellahLaboratory of Heterocyclic Organic Chemistry, Department of Chemistry, Faculty of Sciences, Mohammed V University in RabatDepartment of Chemistry, Tulane UniversityDepartment of Pharmaceutical Chemistry, College of Pharmacy, King Saud UniversityDepartment of Pharmaceutical Chemistry, College of Pharmacy, King Saud UniversityDepartment of Pharmaceutical Sciences, College of Pharmacy, Princess Nourah bint Abdulrahman UniversityAbstract Benzodiazepines are widely recognized for their therapeutic benefits in the treatment of anxiety and insomnia. However, in the pursuit of innovative anticancer agents, they have gained attention as a possible pharmacophore. One of those promising anticancer benzodiazepines is 3b which was demonstrated to exert good antiproliferative effects. To investigate the anticancer effect of 3b, in silico prediction of the possible targets were performed. Then, the predicted targets were investigated through in vitro study. Furthermore, 3b was evaluated for its effects on cell cycle suppression and induction of apoptosis. Molecular docking was used to study the possible types of interactions while molecular dynamics simulations were conducted to estimate the protein–ligand complex’s stability and dynamic behavior. Results demonstrated that 3b is a potent dual inhibitor of HER2 and HDAC1 with IC50 values of 0.023 and 0.041 nM, respectively. Moreover, 3b was found to suppress cell cycle progression in G2/M phase and induce early and late apoptosis in HepG2 cancer cells. Further analysis of apoptotic markers revealed an induction of Caspase 3 and BAX proapoptotic proteins along with a suppression of the antiapoptotic protein (Bcl-2). Molecular docking of 3b into the active site of HER2 and HDAC1 displayed significant types of interactions with active sites of these target proteins while molecular dynamics simulations demonstrated the overall structural stability of HER2 and HDAC1 is maintained or even enhanced upon ligand binding. In conclusion, 3b is a powerful anticancer agent that exerts its effects by inhibiting HDAC1 and HER2, resulting in cell cycle arrest and cancer cell death through apoptosis. Nonetheless, additional investigations are needed to explore its mechanisms and therapeutic efficacy in more detail.https://doi.org/10.1038/s41598-025-97786-7BenzodiazepineHDAC inhibitor, HER2 inhibitorCell cycle analysisProgramed cell deathMolecular dynamics simulations
spellingShingle Nada S. Al-Theyab
Ahmad H. Bakheit
Sanae Lahmidi
Mohammed Mufadhe Alanazi
Awadh M Ali
Khalil Azzaoui
El Mokhtar Essassi
Joel T. Mague
Mohamad Hefnawy
Mohammed Mater Alanazi
Ashwag S. Alanazi
In Silico and in vitro evaluation of the anticancer effect of a 1,5-Benzodiazepin-2-One derivative (3b) revealing potent dual inhibition of HER2 and HDAC1
Scientific Reports
Benzodiazepine
HDAC inhibitor, HER2 inhibitor
Cell cycle analysis
Programed cell death
Molecular dynamics simulations
title In Silico and in vitro evaluation of the anticancer effect of a 1,5-Benzodiazepin-2-One derivative (3b) revealing potent dual inhibition of HER2 and HDAC1
title_full In Silico and in vitro evaluation of the anticancer effect of a 1,5-Benzodiazepin-2-One derivative (3b) revealing potent dual inhibition of HER2 and HDAC1
title_fullStr In Silico and in vitro evaluation of the anticancer effect of a 1,5-Benzodiazepin-2-One derivative (3b) revealing potent dual inhibition of HER2 and HDAC1
title_full_unstemmed In Silico and in vitro evaluation of the anticancer effect of a 1,5-Benzodiazepin-2-One derivative (3b) revealing potent dual inhibition of HER2 and HDAC1
title_short In Silico and in vitro evaluation of the anticancer effect of a 1,5-Benzodiazepin-2-One derivative (3b) revealing potent dual inhibition of HER2 and HDAC1
title_sort in silico and in vitro evaluation of the anticancer effect of a 1 5 benzodiazepin 2 one derivative 3b revealing potent dual inhibition of her2 and hdac1
topic Benzodiazepine
HDAC inhibitor, HER2 inhibitor
Cell cycle analysis
Programed cell death
Molecular dynamics simulations
url https://doi.org/10.1038/s41598-025-97786-7
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