Fluorinated isoindolinone-amino acid hybrids possessing antiproliferative and antitumor activity in vitro and in vivo
Related Experiment Video
Updated: Aug 6, 2026
A Fluorescence-based Protocol for Preliminary Screening of Protein Synthesis Inhibitors from Natural Sources
Published on: January 27, 2026
Soma Mandal1, Rajat Choudhary1, A A I Parvaj Laskar1
1School of Biological Sciences, National Institute of Science Education and Research, HBNI, Jatni, Odisha 752050, India; Homi Bhabha National Institute, Training School Complex, Anushakti Nagar, Mumbai 400094, India.
Abstract:
The limited efficacy and resistance associated with current anticancer therapies necessitate the development of novel agent, particularly for aggressive malignancies. In this study, a series of fluorine-substituted isoindolinone-amino acid conjugates was designed and evaluated for antiproliferative activity using a phenotypic screening approach. A library of 24 compounds was screened against lung, breast and skin cancer cell lines, among which 11 derivatives demonstrated promising antiproliferative activity with IC50 values ranging from 7.36 to 41.99 µM. SAR analysis revealed that incorporation of trifluoromethoxy (-OCF3) substitution with Trp, Tyr, and Phe conjugation significantly enhances activity relative to trifluoromethyl (-CF3) and mono-fluoro analogues. The lead compounds 9c, 9d, 9e, and 10c markedly inhibited cancer cell proliferation, clonogenic survival, and migration, while inducing apoptosis and a pronounced S-phase cell-cycle arrest. Network pharmacology identified CDK2 and GSK3B as key hub genes associated with cell-cycle regulation. Mechanistic studies demonstrated downregulation of CDK2 and PCNA, activation of p53-p21 signaling pathway, and increased γ-H2AX expression, indicating replicating-associated damage. Molecular docking predicted favorable interactions with the CDK2 catalytic pocket, which was further validated by an in vitro CDK2/CyclinA2 kinase inhibition and reduced CDK2 protein expression determined by western blot analysis, collectively implicating CDK2 as a key molecular target. Furthermore, compounds 9c, 9d, 9e and 10c exhibited significant tumor growth inhibition (TGI) values of 53.50%, 79.47%, 64.50% and 69.14% respectively, in the 4T1 murine breast cancer model without evident systemic toxicity. Collectively, these findings identify fluorinated isoindolinone-amino acid conjugates as promising anticancer leads targeting CDK2-associated cell-cycle signaling against triple-negative breast cancer.
Insights
New fluorinated isoindolinone-amino acid conjugates show potent anticancer activity. These novel agents target cell-cycle regulation via CDK2, demonstrating significant tumor growth inhibition in preclinical models.
Area of Science:
- Medicinal Chemistry
- Pharmacology
- Molecular Biology
Background:
- Current anticancer therapies face limitations in efficacy and drug resistance, especially for aggressive cancers.
- Novel therapeutic agents are urgently needed to overcome these challenges.
- Fluorine substitution in drug design can modulate pharmacokinetic and pharmacodynamic properties.
Purpose of the Study:
- To design and evaluate novel fluorine-substituted isoindolinone-amino acid conjugates for antiproliferative activity.
- To identify lead compounds and elucidate their mechanism of action against cancer cell lines.
- To assess the in vivo efficacy and safety of promising candidates in a preclinical cancer model.
Main Methods:
- Phenotypic screening of 24 synthesized compounds against lung, breast, and skin cancer cell lines.
- Structure-activity relationship (SAR) analysis to optimize fluorine substitution and amino acid conjugation.
- In vitro assays to evaluate effects on cell proliferation, apoptosis, cell-cycle arrest, and migration.
- Network pharmacology and molecular docking to identify molecular targets (e.g., CDK2).
- In vivo studies using the 4T1 murine breast cancer model to assess tumor growth inhibition and toxicity.
Main Results:
- Eleven compounds exhibited promising antiproliferative activity (IC50: 7.36–41.99 µM).
- Trifluoromethoxy (-OCF3) substitution with Trp, Tyr, and Phe enhanced activity.
- Lead compounds (9c, 9d, 9e, 10c) inhibited proliferation, survival, migration, induced apoptosis, and caused S-phase arrest.
- Mechanism involved CDK2/GSK3B targeting, p53-p21 activation, and DNA damage (γ-H2AX).
- Compounds showed significant tumor growth inhibition (up to 79.47%) in vivo with no apparent toxicity.
Conclusions:
- Fluorinated isoindolinone-amino acid conjugates are effective anticancer agents.
- CDK2 is identified as a key molecular target mediating the observed antiproliferative effects.
- These compounds represent promising leads for developing novel therapies against triple-negative breast cancer and other malignancies.
Related Concept Videos
02:50Combination Therapies and Personalized Medicine
The combination of the drug acetazolamide and sulforaphane is a good example of combination therapy to treat cancer. The cells in the interior of a large tumor often die due to the hypoxic and...
02:07FISH - Fluorescent In-situ Hybridization
01:28Inhibitors of Bacterial DNA Synthesis
02:57Targeted Cancer Therapies
There are several types of targeted therapies against specific...
01:22Antiviral Nucleoside Inhibitors