Page 208 - EJMO-9-3

Eurasian Journal of
Medicine and Oncology FN3K–Nrf2 axis inhibition in breast cancer

interactions. This approach facilitates rational drug offering promising prospects for advancing personalized
design by streamlining lead identification from extensive treatment modalities. 4
chemical libraries, minimizing experimental workload,
and improving hit enrichment. Lead molecule selection 2. Materials and methods
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for therapeutic development represents a critical phase in 2.1. SBVS
the drug discovery pipeline, necessitating the validation of
computational predictions through robust experimental 2.1.1. Materials
assays. Lead candidates were strategically selected 2.1.1.1. Software
from the initial SBVS hits by evaluating their predicted Schrödinger Maestro (Schrödinger, LLC, USA) was used
binding strength and structural compatibility with the as a comprehensive molecular modeling environment
FN3K enzyme. These candidates were then subjected to for preparing, visualizing, and analyzing molecular
biological evaluation to confirm their ability to inhibit structures. For molecular docking, the Schrödinger Glide
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FN3K enzymatic activity. The overarching aim was to
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identify potent inhibitors capable of suppressing FN3K module within Maestro, was employed, as it is widely used
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function, thereby offering potential therapeutic benefit in for VS and drug discovery.
the context of breast cancer management. 2.1.1.2. Datasets
To confirm computational findings and evaluate the Kinase inhibitors approved by the FDA are integral
biological significance of the selected FN3K inhibitors, components of cancer treatment due to their targeted
in vitro assays were performed using quantitative PCR mechanisms of action. The WHO Model Essential
12
(qPCR) and Western blotting. These assays were utilized Medicines list comprises pharmacological agents
to investigate the expression patterns of FN3K and the recognized for their clinical efficacy, safety profile, and
redox-responsive transcription factor Nrf2, a key regulator widespread use in medical practice, as endorsed by the
of cellular antioxidant defenses and chemoresistance. WHO. In addition, FDA-approved anti-breast cancer
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Earlier research has shown that abnormal elevation of agents represent a curated list of approved drugs specifically
Nrf2 expression promotes tumor cell survival in oxidative targeting breast cancer, offering potential repositioning
environments, while its suppression increases cancer opportunities. 14
cell vulnerability to chemotherapeutic agents. This study
also investigated the relationship between FN3K and 2.1.1.3. Computational resources
Nrf2 expression across various breast cancer subtypes, A high-performance computing cluster or a powerful
underscoring their potential co-regulatory involvement workstation with sufficient memory and processing power
in redox homeostasis and tumor progression. By was used to handle molecular simulations and docking
7,17
identifying compounds that selectively inhibit FN3K calculations. 20
activity, we aimed to uncover novel therapeutic agents
with potential applications in breast cancer management. 2.1.1.4. Protein and ligand data
FN3K plays a pivotal role in cancer progression by The three-dimensional (3D) structure of the target
stabilizing and enhancing the oncogenic potential of protein was generated through comparative or homology
Nrf2 through deglycation, thereby enabling sustained modeling to enable precise molecular docking and reliable
activation of antioxidant and survival pathways in tumor assessment of ligand–protein interactions. Ligand
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cells. Therefore, targeting FN3K offers a promising datasets were obtained from reputable sources, such as
therapeutic strategy to attenuate aberrant Nrf2 signaling, DrugBank, PubChem, and the FDA, providing well-
overcome drug resistance, and sensitize cancer cells to characterized small molecules for VS. 12-14
oxidative stress. The identification of promising lead
compounds exhibiting strong FN3K-inhibitory activity 2.1.2. Methods
lays the groundwork for the rational design of novel anti- 2.1.2.1. Construction of the 3D structure of the protein
cancer strategies, particularly in breast cancer, where
FN3K overexpression has been linked to unfavorable The 3D structure of human FN3K was constructed through
clinical outcomes. Taken together, this work illustrates a systematic multistep process. First, the amino acid
the effectiveness of combining computational approaches, sequence of human FN3K, consisting of 309 residues and
such as VS, with in vitro validation to accelerate the indexed under UniProt ID Q9H479, was retrieved from the
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identification of targeted therapeutic candidates. The UniProt database. Phylogenetic similarities among FN3K
FN3K–Nrf2 signaling axis emerges as a novel and relatively sequences were then assessed using Clustal Omega, a robust
unexplored target in cancer research, with FN3K inhibition tool for multiple sequence alignment. For homology
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Volume 9 Issue 3 (2025) 200 doi: 10.36922/EJMO025150114