2-(4-Nitrophenyl)isothiazol-3(2H)-one - A Promising Selective Agent against Hepatocellular Carcinoma Cells

Sep 21, 2026

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Liver cancer ranks among the most prevalent malignancies worldwide and remains a leading cause of cancer-related death. Numerous isothiazolone derivatives have been explored as anticancer agents, yet evidence on their activity against liver cancer is limited. Two nitrophenyl-isothiazolones were preliminarily evaluated for cytotoxicity on hepatoma Huh7 cells and on immortalized human hepatocytes as a non-cancerous model. IsoB, obtained from IsoA by removing the benzoyl group, produced the strongest, time-dependent cytotoxicity in Huh7 cells and showed selective toxicity versus IHH cells. IsoB also outperformed or contrasted with 5-fluorouracil, which did not increase toxicity in the liver cancer models. Mechanistic studies indicated reduced mitochondrial membrane potential and increased mitochondrial superoxide after treatment, while RT-qPCR suggested apoptosis-mediated cell death, with TP53 upregulation and MYCN downregulation.

LanguageEnglish

CategoryResearch & Report

FormatPDF

Pages20

Reading Time50 min

AccessFree to download

Introduction

Isothiazolones as anticancer agents

Abstract

Study rationale and models

Compounds and cytotoxicity results

Selectivity and comparison with 5-FU

Mitochondrial effects and mechanism (RT-qPCR)

pharmaceuticals Article 2-(4-Nitrophenyl)isothiazol-3(2H)-one: A Promising Selective Agent against Hepatocellular Carcinoma Cells Sofia Marka 1, *, Maria-Eleftheria Zografaki 2, Georgia Tsolomiti 1, Katerina I. Kalliampakou 2,3, Athanasios Tsolomitis 4, Christina Koumantou 1, Despina Smirlis 5, Niki Vassilaki 3 and Spyros Kintzios 1, * Citation: Marka, S.; Zografaki, M.-E.; Tsolomiti, G.; Kalliampakou, K.I.; Tsolomitis, A.; Koumantou, C.; Smirlis, D.; Vassilaki, N.; Kintzios, S. 2-(4-Nitrophenyl)isothiazol-3(2H)-one: A Promising Selective Agent against Hepatocellular Carcinoma Cells. Pharmaceuticals 2024, 17, 673 . [https://doi.org/10.3390/ph17060673](https://doi.org/10.3390/ph17060673) Academic Editor: Emmanuel Moreau Received: 15 March 2024 Revised: 10 May 2024 Accepted: 20 May 2024 Published: 24 May 2024 Copyright: © 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ([https://](https://)[ ](https://)[creativecommons.org/licenses/by/](creativecommons.org/licenses/by/)[ ](creativecommons.org/licenses/by/)[4.0/](4.0/)) . 1 Laboratory of Cell Technology, Department of Biotechnology, School of Applied Biology and Biotechnology, Agricultural University of Athens, 11855 Athens, Greece; georgia.tsolomiti@aua.gr (G.T.); [christinakoumantou@gmail.com](christinakoumantou@gmail.com) (C.K.) 2 Laboratory of Molecular Biology, Department of Biotechnology, School of Applied Biology and Biotechnology, Agricultural University of Athens, 11855 Athens, Greece; mzografaki@aua.gr (M.-E.Z.); [kalliamp@aua.gr](kalliamp@aua.gr) (K.I.K.) 3 Laboratory of Molecular Virology, Hellenic Pasteur Institute, 11521 Athens, Greece; [nikiv@pasteur.gr](nikiv@pasteur.gr) 4 School of Chemical Engineering, National Technical University, 15772 Athens, Greece; [athtsolom@gmail.com](athtsolom@gmail.com) 5 Molecular Parasitology Laboratory, Microbiology Department, Hellenic Pasteur Institute, 11521 Athens, Greece; [penny@pasteur.gr](penny@pasteur.gr) * Correspondence: [smarka@aua.gr](smarka@aua.gr) (S.M.); [skin@aua.gr](skin@aua.gr) (S.K.); Tel.: +30-2105294292 (S.M. & S.K.) Abstract: Liver cancer ranks among the most prevalent malignancies globally and stands as a leading cause of cancer-related mortality. Numerous isothiazolone derivatives and analogues have been synthesized and investigated for their potential as anticancer agents; however, limited data exist regarding their efficacy against liver cancer. In the present study, two nitrophenyl-isothiazolones, the 5-benzoyl-2-(4-nitrophenyl)isothiazol-3(2H)-one (IsoA) and the 2-(4-nitrophenyl)isothiazol-3(2H)-one (IsoB), were preliminarily investigated for their cytotoxicity against hepatoma human (Huh7) cells asa liver cancer model and Immortalized Human Hepatocytes (IHHs) as a model of non-cancerous hepatocytes. IsoB, derived from IsoA after removal of the benzoyl moiety, demonstrated the highest cytotoxic effect against Huh7 cells with CC 50 values of 19.3 µM at 24 h, 16.4 µM at 48 h, and 16.2 µM at 72 h of incubation, respectively. IsoB also exhibited selective toxicity against the liver cancerous Huh7 cells compared to IHH cells, reinforcing its role as a potent and selective anticancer agent. Remarkably, the cytotoxicity of IsoB was higher when compared with the standard chemotherapeutical agent 5-fluorouracil (5-FU), which also failed to exhibit higher toxicity against the liver cancerous cell lines. Moreover, IsoB-treated Huh7 cells presented a noteworthy reduction in mitochondrial membrane potential (∆Ψm) after 48 and 72 h, while mitochondrial superoxide levels showed an increase after 24 h of incubation. The molecular mechanism of the IsoB cytotoxic effect was also investigated using RT-qPCR, revealing an apoptosis-mediated cell death along with tumor suppressor TP53 overexpression and key-oncogene MYCN downregulation. Keywords:

FAQs

What problem does the study address regarding liver cancer treatment?

Liver cancer is a major cause of cancer mortality, but limited data exist on how isothiazolone derivatives perform against liver cancer. The study evaluates whether specific nitrophenyl-isothiazolones can act as anticancer agents against hepatocellular carcinoma models.

How do IsoA and IsoB differ, and which one is more effective?

IsoB is derived from IsoA after removal of the benzoyl moiety. IsoB shows the highest cytotoxic effect against Huh7 hepatoma cells across 24, 48, and 72 hours.

What mechanism is proposed for IsoB’s cytotoxic effect?

IsoB-treated Huh7 cells display reduced mitochondrial membrane potential and increased mitochondrial superoxide. RT-qPCR results support apoptosis-mediated cell death, with TP53 overexpression and MYCN downregulation.

Sep 21, 2026

Uploaded by Violet