# Integrated In Silico and In Vitro Study of Copper Nanocatalyzed Carbonyl‐Functionalized Triazoles—Inducing S Phase Cell Cycle Arrest and Apoptosis in MCF‐7

## TL;DR

This study identifies two new copper-based compounds that effectively target breast cancer cells by causing cell cycle arrest and apoptosis.

## Contribution

The discovery of copper nanocatalyzed triazoles (3i and 3j) as potent inducers of apoptosis in breast cancer cells through modulation of BAX/BCL2 and mitochondrial dysfunction.

## Key findings

- Compounds 3i and 3j showed high selectivity indices (2.30 and 4.44) and strong binding to BAX and BCL2 proteins.
- The compounds caused S-phase cell cycle arrest and apoptosis in MCF-7 cells via ROS-mediated glutathione depletion and mitochondrial dysfunction.
- Molecular dynamics simulations confirmed the stability of protein-ligand complexes for the lead compounds.

## Abstract

The demand for novel, selective anticancer agents, driven by drug resistance and systemic toxicity of current treatments, underscores the importance of targeted drug discovery. Present research involved cytotoxic screening of a series of synthesized copper nanocatalyzed carbonyl‐functionalized triazoles (3a
–
p), where 3i and 3j have shown highest selectivity index (SI) scores of 2.30 and 4.44, respectively. Computational validation of the lead compounds demonstrated specific interaction with BCL2‐associated X protein (BAX) and BCL2, characterized by strong binding affinities ranging between −6.73 and −7.70 kcal/mol. Corresponding protein–ligand complexes demonstrated robust conformational stability throughout their 100 ns of molecular dynamics simulation. Subsequent in vitro validation using MCF‐7 cells firmly corroborated the in silico findings, by revealing significant upregulation of BAX (p < 0.001) and downregulation of BCL2 (p < 0.001). Compound induced cellular stress, elevated the ROS‐producing cell population up to 40%. Resulting cellular oxidative stress, rapidly depleted the glutathione reserves up to 50% (p < 0.001), consequently compromising the mitochondrial membrane potential leading to mitochondrial dysfunction. Furthermore, the compound induced S‐phase cell cycle arrest (upto 51.5%), played a pivotal role in promoting apoptosis by activating DNA damage response pathways. In conclusion, this study has successfully identified two lead compounds (3i & 3j) that modulate multiple converging oncogenic pathways, providing compelling preclinical candidates for targeted management of breast cancer.

A targeted drug discovery pipeline identified novel copper‐nanocatalyzed carbonyl‐functionalized triazoles as potent breast cancer therapeutics (selectivity index up to 4.44). Lead compounds (3i, 3j) induced apoptosis in breast cancer cells (MCF‐7) by modulating BAX/BCL2, causing reactive oxygen species (ROS)‐mediated glutathione (GSH) depletion, mitochondrial membrane potential (MMP) compromise, and DNA damage response activation, thereby causing S‐phase cell cycle arrest.© 2026 WILEY‐VCH GmbH