Design, Synthesis, and Cytotoxicity Evaluation of Novel Indolin-2-One Based Molecules on Hepatocellular Carcinoma HepG2 Cells as Protein Kinase Inhibitors

A series of indolinone-based derivatives were designed and synthesized using the hybrid pharmacophoric design approach as cytotoxic kinase inhibitors. The cytotoxic effects of the designed molecules were tested against MCF-7 and HepG-2 cell lines. Compounds <b>9</b> and <b>20</b...

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Main Authors: Manal M. Kandeel, Mohamed Kamal AbdElhameid, Mohamed Adel, Muhammad Y. Al-Shorbagy, Ahmed T. Negmeldin
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Molecules
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Online Access:https://www.mdpi.com/1420-3049/30/5/1105
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author Manal M. Kandeel
Mohamed Kamal AbdElhameid
Mohamed Adel
Muhammad Y. Al-Shorbagy
Ahmed T. Negmeldin
author_facet Manal M. Kandeel
Mohamed Kamal AbdElhameid
Mohamed Adel
Muhammad Y. Al-Shorbagy
Ahmed T. Negmeldin
author_sort Manal M. Kandeel
collection DOAJ
description A series of indolinone-based derivatives were designed and synthesized using the hybrid pharmacophoric design approach as cytotoxic kinase inhibitors. The cytotoxic effects of the designed molecules were tested against MCF-7 and HepG-2 cell lines. Compounds <b>9</b> and <b>20</b> were the most cytotoxic, with IC<sub>50</sub> values against HepG-2 and MCF-7 cells ranging from 2.53 to 7.54 µM. Additionally, compounds <b>9</b> and <b>20</b> were also found to be slightly more cytotoxic than indirubin with 2.2–2.7-fold higher cytotoxicity with HepG-2 cells. CDK-2 and CDK-4 kinase enzyme inhibition assay showed that compound <b>9</b> had a higher inhibitory effect (4.8-fold) than indirubin against CDK-2 and comparable inhibition against CDK-4. Moreover, compound <b>20</b> displayed nanomolar inhibitory action against both EGFR kinase and VFGFR-2 enzyme, which were around 8.8- and 5.4-fold higher than the IC<sub>50</sub> values of indirubin. Compounds <b>9</b> and <b>20</b> induced cell cycle arrest at the G<sub>1</sub> phase on HepG2 cells. The levels of the key apoptotic proteins assessed revealed elevated levels of the Bax/Bcl-2 ratio, which in turn initiated the caspase3/7 cascade that led to the activation of both intrinsic and extrinsic apoptotic pathways. The cell cycle inhibitory proteins p53 and p21 were significantly upregulated upon treatment with compounds <b>9</b> and <b>20</b>. The docking results revealed that compound <b>9</b> exhibits stronger binding affinity to CDK-2 than indirubin, and compound <b>20</b> showed a similar binding mode to sorafenib with VEGFR-2.
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spelling doaj-art-74413a523f5c4e24b01ec44d837861a12025-08-20T02:06:12ZengMDPI AGMolecules1420-30492025-02-01305110510.3390/molecules30051105Design, Synthesis, and Cytotoxicity Evaluation of Novel Indolin-2-One Based Molecules on Hepatocellular Carcinoma HepG2 Cells as Protein Kinase InhibitorsManal M. Kandeel0Mohamed Kamal AbdElhameid1Mohamed Adel2Muhammad Y. Al-Shorbagy3Ahmed T. Negmeldin4Department of Pharmaceutical Organic Chemistry, Faculty of Pharmacy, Cairo University, Cairo 11562, EgyptDepartment of Pharmaceutical Organic Chemistry, Faculty of Pharmacy, Cairo University, Cairo 11562, EgyptDepartment of Pharmaceutical Organic Chemistry, Faculty of Pharmacy, Egyptian Russian University, Cairo 11829, EgyptDepartment of Pharmaceutical Sciences, College of Pharmacy, Gulf Medical University, Ajman 4184, United Arab EmiratesDepartment of Pharmaceutical Organic Chemistry, Faculty of Pharmacy, Cairo University, Cairo 11562, EgyptA series of indolinone-based derivatives were designed and synthesized using the hybrid pharmacophoric design approach as cytotoxic kinase inhibitors. The cytotoxic effects of the designed molecules were tested against MCF-7 and HepG-2 cell lines. Compounds <b>9</b> and <b>20</b> were the most cytotoxic, with IC<sub>50</sub> values against HepG-2 and MCF-7 cells ranging from 2.53 to 7.54 µM. Additionally, compounds <b>9</b> and <b>20</b> were also found to be slightly more cytotoxic than indirubin with 2.2–2.7-fold higher cytotoxicity with HepG-2 cells. CDK-2 and CDK-4 kinase enzyme inhibition assay showed that compound <b>9</b> had a higher inhibitory effect (4.8-fold) than indirubin against CDK-2 and comparable inhibition against CDK-4. Moreover, compound <b>20</b> displayed nanomolar inhibitory action against both EGFR kinase and VFGFR-2 enzyme, which were around 8.8- and 5.4-fold higher than the IC<sub>50</sub> values of indirubin. Compounds <b>9</b> and <b>20</b> induced cell cycle arrest at the G<sub>1</sub> phase on HepG2 cells. The levels of the key apoptotic proteins assessed revealed elevated levels of the Bax/Bcl-2 ratio, which in turn initiated the caspase3/7 cascade that led to the activation of both intrinsic and extrinsic apoptotic pathways. The cell cycle inhibitory proteins p53 and p21 were significantly upregulated upon treatment with compounds <b>9</b> and <b>20</b>. The docking results revealed that compound <b>9</b> exhibits stronger binding affinity to CDK-2 than indirubin, and compound <b>20</b> showed a similar binding mode to sorafenib with VEGFR-2.https://www.mdpi.com/1420-3049/30/5/1105HepG-2 cellsMCF-7isatinquinazolinesprotein kinasestyrosine kinase
spellingShingle Manal M. Kandeel
Mohamed Kamal AbdElhameid
Mohamed Adel
Muhammad Y. Al-Shorbagy
Ahmed T. Negmeldin
Design, Synthesis, and Cytotoxicity Evaluation of Novel Indolin-2-One Based Molecules on Hepatocellular Carcinoma HepG2 Cells as Protein Kinase Inhibitors
Molecules
HepG-2 cells
MCF-7
isatin
quinazolines
protein kinases
tyrosine kinase
title Design, Synthesis, and Cytotoxicity Evaluation of Novel Indolin-2-One Based Molecules on Hepatocellular Carcinoma HepG2 Cells as Protein Kinase Inhibitors
title_full Design, Synthesis, and Cytotoxicity Evaluation of Novel Indolin-2-One Based Molecules on Hepatocellular Carcinoma HepG2 Cells as Protein Kinase Inhibitors
title_fullStr Design, Synthesis, and Cytotoxicity Evaluation of Novel Indolin-2-One Based Molecules on Hepatocellular Carcinoma HepG2 Cells as Protein Kinase Inhibitors
title_full_unstemmed Design, Synthesis, and Cytotoxicity Evaluation of Novel Indolin-2-One Based Molecules on Hepatocellular Carcinoma HepG2 Cells as Protein Kinase Inhibitors
title_short Design, Synthesis, and Cytotoxicity Evaluation of Novel Indolin-2-One Based Molecules on Hepatocellular Carcinoma HepG2 Cells as Protein Kinase Inhibitors
title_sort design synthesis and cytotoxicity evaluation of novel indolin 2 one based molecules on hepatocellular carcinoma hepg2 cells as protein kinase inhibitors
topic HepG-2 cells
MCF-7
isatin
quinazolines
protein kinases
tyrosine kinase
url https://www.mdpi.com/1420-3049/30/5/1105
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AT mohamedadel designsynthesisandcytotoxicityevaluationofnovelindolin2onebasedmoleculesonhepatocellularcarcinomahepg2cellsasproteinkinaseinhibitors
AT muhammadyalshorbagy designsynthesisandcytotoxicityevaluationofnovelindolin2onebasedmoleculesonhepatocellularcarcinomahepg2cellsasproteinkinaseinhibitors
AT ahmedtnegmeldin designsynthesisandcytotoxicityevaluationofnovelindolin2onebasedmoleculesonhepatocellularcarcinomahepg2cellsasproteinkinaseinhibitors