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Page 417 Aydin et al. J Transl Genet Genom. 2025;9:406-26 https://dx.doi.org/10.20517/jtgg.2025.108

repositioning appears to be a promising strategy because it offers a unique way to identify new effects of
approved drugs more quickly, at lower cost, and with shorter clinical trial periods. This approach has been
[85]
and continues to be applied to a wide range of diseases .

In this study, DMD was evaluated by integrating transcriptome-level data with network biology approaches
to identify new drug candidates, including celastrol, radicicol, apigenin triacetate, emetine dihydrochloride
hydrate, and withaferin-A, which have been suggested as potential therapeutics through in silico validations
for DMD management. The construction of DMD-specific biological networks at three molecular levels -
TFs, miRNAs, and proteins - allowed us to assess the most essential hubs of the disease that carry the
information flow potentially responsible for pathogenesis [Figure 2A-C]. These 33 hubs were defined as
“network signatures of DMD” and further evaluated for their predictive performance in distinguishing
healthy from disease samples using PCA. They demonstrated significant effects in clustering healthy
samples and DMD patients, with 100% sensitivity and 100% specificity [Table 3 and Figure 3]. Network
signature-based drug repositioning revealed drugs with the potential to reverse gene expression patterns
[Table 4]. We selected approved or investigational drugs, including celastrol, apigenin triacetate, emetine
dihydrochloride hydrate, withaferin-A, dipyridamole, and radicicol. Some of these compounds were
reported to have antineoplastic effects [Table 4]. All these small molecules are discussed comprehensively in
subsequent sections.

[86]
Radicicol (monorden) is isolated from the fungus Monosporium bonorden and is known as an antibiotic .
Radicicol, a powerful sedative with a low toxicity rate, was first identified as a protein tyrosine kinase
[75]
inhibitor, and later its heat shock protein 90 (Hsp90) inhibitory activity was revealed . Since Hsp90 affects
the growth of many oncogenic proteins, its inhibition leads to the shutdown of multiple pathways involved
in cancer [87,88] . As a result of drug repositioning research, radicicol became one of 19 drugs that provide rapid
inhibition against Naegleria fowleri . Radicicol has also been reported as a repositioned drug candidate for
[89]
breast cancer and coronavirus disease 2019 (COVID-19) [90,91] .

As an ayurvedic herb, Ashwagandha (Withania somnifera) contains a steroidal lactone called withaferin-A.
It is well-known for its anti-inflammatory, anti-tumorigenic, and anti-carcinogenic activities [92-94] . It is
typically utilized for patients with other unexplained gynecological malignancies, as well as lung, breast,
ovarian, colorectal, prostate, gastrointestinal, cervical, and hematological cancers [93,94] . Beyond these diseases,
withaferin-A has demonstrated potential as a treatment for conditions such as osteoarthritis, Parkinson’s
disease, Alzheimer’s disease, and cystic fibrosis . By preventing nuclear factor-κB (NF-κB) activation and
[95]
disrupting the CDC37-Hsp90 complex, withaferin-A has the potential to be used as a therapeutic drug in
the treatment of cancer, hematological malignancies, and chronic inflammatory disorders . Overactivity of
[96]
NF-κB results in the loss and disruption of muscle cells, whereas withaferin-A-induced NF-κB suppression
can promote the development of new myofibers and may be useful in DMD treatment . Currently, no
[97]
study in the literature has reported DMD treatment with withaferin-A.

Apigenin, naturally produced by Thymelaeaceae, Verbenaceae, and Selaginellaceae plants, is a flavonoid
component . It exhibits anti-spasmodic, anti-inflammatory, antioxidant, anti-cancer (osteosarcoma,
[98]
prostate, bladder, liver, and colorectal cancer cells), and antiproliferative properties [99,100] . Apigenin also has
strong antiviral activity, as it inhibits Mpro, the primary SARS-CoV-2 (severe acute respiratory syndrome
[101]
coronavirus 2) protease, thereby blocking the virus’s ability to replicate . By modulating integrin signaling
pathways and downregulating Signal transducer and activator of transcription 3 (STAT3) target genes
(TWIST1, MMP-2, MMP-9, and VEGF), it inhibits melanoma cell motility and invasion, and it also exhibits
[102]
an anti-metastatic effect by lowering VEGF gene expression in melanoma cells . In a recent study,