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Article Open Access

Identification of non‑oncology drugs with activity against inflammatory and triple‑negative breast cancer cell lines

  • Authors:
    • Syed Ahmad
    • Sara Kandil
    • Kevin P. Williams
    • John E. Scott
  • View Affiliations / Copyright

    Affiliations: Department of Pharmaceutical Sciences, Biomanufacturing Research Institute and Technology Enterprise, North Carolina Central University, Durham, NC 27707, USA
    Copyright: © Ahmad et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 75
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    Published online on: December 15, 2025
       https://doi.org/10.3892/ol.2025.15428
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Abstract

Inflammatory breast cancer (IBC) and triple‑negative breast cancer (TNBC) are subtypes of breast cancer with the lowest 5‑year survival rates. One potential approach to increase survival rates for these cancers may be to identify tolerable and inexpensive daily maintenance therapies that could be administered in between standard of care treatments. The present study aimed to identify non‑oncology drugs (NODs) that show activity against IBC/TNBC that could be repurposed for this use. Thus, a collection of NODs were screened and curated from a published database using cell line SUM‑149, which is both an IBC and TNBC cell line (TN‑IBC). In a parallel screening approach, a subset of these NODs were tested for activity against two other TNBC cell lines. Together, 15 drugs with activity against this IBC cell line were identified and grown as spheroids. A number of these drugs have not been previously identified as having activity in an IBC cell line. In addition, eight of these were understudied for TNBC and, therefore, they were tested against three TNBC cell lines grown as spheroids, resulting in variable breadth of activities. Drug potency (GI50) was also assessed relative to their Cmax reported in human studies. Eltrombopag, a thrombocytopenia drug, was active against SUM‑149 and two other TNBC cell lines with a GI50:Cmax ratio of ~1. Papaverine, a vasodilator, was also active against SUM‑149 and one other TNBC cell line with a GI50:Cmax ratio of ~2. Mozavaptan, a hyponatremia drug, inhibited proliferation in 4 out of 5 cell lines (all except SUM‑149) with GI50=2.7‑7.6 µM (Cmax unknown). Spironolactone (GI50:Cmax=10) had anti‑proliferative activity only against SUM‑149, suggesting a unique vulnerability. Overall, our data indicated that eltrombopag, papaverine and other identified NODs may have potential for being repurposed for TN‑IBC and TNBC in general.
View Figures

Figure 1

Summary of selection and screening
method for identifying drugs with potential for repurposing. A
search of the published PRISM database for active drugs in at least
three TNBC cell lines resulted in 100 candidates. After drugs were
eliminated from this list due to a variety of reasons that reduced
potential for rapid repurposing (for example, only approved in
animals, only approved for topical use), 48 drugs remained for
testing. These were tested in a primary single concentration screen
(10 µM) and subsequent actives were then tested in concentration
response using the TN-IBC SUM-149 cell line grown as spheroids
resulting in 15 confirmed active drugs. A subset of 33 drugs that
have been understudied for TNBC were tested in a single
concentration (10 µM) screen using two TNBC cell lines in a 2-D
cell proliferation assay followed by determination of
GI50 and cytotoxicity for actives using three TNBC cell
lines also grown in 2-D. Five confirmed actives from the 2-D
screening were combined with the actives from the SUM-149 screen
and assessed for literature reports related to TNBC and IBC
resulting in eight understudied drugs. These were profiled for
GI50 against a total of four TNBC cells lines (including
SUM-149) grown as spheroids. TNBC, triple-negative breast cancer;
TN-IBC, triple-negative inflammatory breast cancer; 2-D,
two-dimension; IBC, inflammatory breast cancer; GI, growth
inhibition.

Figure 2

Multiparametric screen of selected
NODs for activity against TN-IBC cell line SUM-149 grown in
spheroid culture. A total of 48 drugs (as indicated on the y-axis)
were tested at 10 µM against SUM-149 grown as spheroids. DMSO wells
were solvent control wells used to normalize the data and
paclitaxel and cycloheximide were used as control inhibitors. (A)
After 96 h of drug exposure, CellTiter Glo was used to determine
ATP levels as a reflection of cell viability, calculated as
percentage of controls. Red bars represent drugs that had
significantly reduced cell viability compared to DMSO control using
unpaired t-test and therefore were selected for GI50
determinations (*P<0.05 compared to DMSO). (B) The change in
spheroid area between 0 and 96 h was calculated for each drug as a
percentage of DMSO control. (C) Non-permeable CellTox Green dye was
used to detect free DNA by measuring the total green fluorescence
of the well and normalizing it to controls. Normalized data
>100% was indicative of cytotoxicity. The drug marked
‘Autofluor’ showed fluorescence at time 0 indicating that the
fluorescence was high due to autofluorescence of the drug.
Screening data shown represents the mean ± standard deviation of
the aggregated normalized data from n=2 or 3 independent
experiments where each experiment employed duplicate technical
replicates. NOD, non-oncology drug; TN-IBC, triple-negative
inflammatory breast cancer; DMSO, dimethyl sulfoxide.

Figure 3

2-D screen of selected NODs for
activity against two TNBC cell lines. 32 drugs (as indicated on the
y-axis) were tested at 10 µM against (A) SUM-159 and (B) MDA-MB-231
grown in 2-D. Change in monolayer confluency between 0 and 72 h was
calculated for each drug as a percentage of DMSO control. DMSO
wells were solvent control wells used to normalize the data and
paclitaxel and cycloheximide were used as control inhibitors. Red
bars represent drugs that had significantly reduced cell viability
compared to DMSO control using unpaired t-test and displayed ≥20%
inhibition (*P<0.05 compared to DMSO). These active drugs were
selected for GI50 determinations. Screening data shown
represents the mean ± standard deviation of the aggregated
normalized data from n=3 independent experiments where each
experiment employed duplicate technical replicates. NOD,
non-oncology drug; TNBC, triple-negative breast cancer; DMSO,
dimethyl sulfoxide.

Figure 4

GI50 determinations for
active drugs against SUM-149 and a panel of other TNBC cell lines
grown as spheroids. Relative cell proliferation (as a percentage of
controls) was determined in response to the indicated
concentrations of drugs tested against SUM-149. Selected compounds
were also profiled with an additional three TNBC cell lines,
SUM-159, HCC1806 and MDA-MB-231, as indicated. The concentration
response data was plotted for (A) adefovir, (B) albendazole, (C)
artesunate, (D) bisacodyl, (E) clofazimine, (F) dolutegravir, (G)
dronedarone, (H) eltrombopag, (I) halofantrine, (J) ivermectin, (K)
mozavaptan, (L) mycophenolic acid, (M) niclosamide, (N) papaverine,
(O) pentamidine, (P) spironolactone and (Q) tolvaptan. All cell
lines were grown in spheroid form for this assay with 96 h drug
treatment followed by viable cell number assessed by CellTiter Glo.
Relative cell proliferation (as opposed to total cells) was
calculated by subtracting mean 0-day plate values from all wells
and normalizing to controls such that a 0 value indicated no cell
proliferation during the course of the assay (same number of viable
cells as on day of treatment), while negative values indicated
cytotoxicity (fewer viable cells than on day of drug treatment).
Data shown are representative of n≥3 independent experiments and
the data points and error bars represent the mean ± standard
deviation of triplicate technical replicates. GI, growth
inhibition; TNBC, triple-negative breast cancer.

Figure 5

GI50 determination of
active drugs from 2-D screen against a panel of three TNBC cell
lines. Relative cell proliferation (as a percentage of controls)
and cytotoxicity was determined in response to the indicated
concentrations of drugs tested against SUM-159, HCC1806 and
MDA-MB-231, as indicated. All cell lines were grown in 2-D.
Immediately after drug treatment, confluency and CellTox Green dye
fluorescence (cytotoxicity) was monitored for 72 h with the
Incucyte. Relative cell proliferation was calculated by subtracting
the initial confluency (based the first read) from the final
confluency measurement for each well and normalized to controls.
Therefore, a 0% value indicates no change in confluency compared to
the start of the assay. Relative cell death after 72 h was
calculated by dividing the area of green dye fluorescence by the
total confluency area (based on brightfield imaging) and
multiplying by 100. Separate plots are provided for cell
proliferation and cytotoxicity for (A and B, respectively)
adefovir, (C and D, respectively) eltrombopag, (E and F,
respectively) mozavaptan, (G and H, respectively) mycophenolic
acid, (I and J, respectively) pentamidine and (K and L,
respectively) halofantrine. Data shown are representative of n≥3
independent experiments and the data points and error bars
represent the mean ± standard deviation of triplicate technical
replicates. GI, growth inhibition; TNBC, triple-negative breast
cancer; 2-D, two-dimension.

Figure 6

Graphical representations of mean
GI50 values and GI50:Cmax ratios
for active drugs. The GI50 data from Tables I and III were represented as bar graphs. (A)
Mean GI50 values of compounds tested only against
SUM-149 (as indicated) or against all four cell lines were plotted.
No error bars indicate that the GI50 is greater than the
highest concentration tested and thus shown as equal to that high
concentration (i.e., GI50 of >20 µM is indicated by a
bar at 20 µM without error bars). (B) The mean GI50
values were divided by the Cmax values from Table I and plotted for each indicated
drug. The ratio of one is indicated with a horizontal black line.
GI, growth inhibition; Cmax, maximal concentration.
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Copy and paste a formatted citation
Spandidos Publications style
Ahmad S, Kandil S, Williams KP and Scott JE: Identification of non‑oncology drugs with activity against inflammatory and triple‑negative breast cancer cell lines. Oncol Lett 31: 75, 2026.
APA
Ahmad, S., Kandil, S., Williams, K.P., & Scott, J.E. (2026). Identification of non‑oncology drugs with activity against inflammatory and triple‑negative breast cancer cell lines. Oncology Letters, 31, 75. https://doi.org/10.3892/ol.2025.15428
MLA
Ahmad, S., Kandil, S., Williams, K. P., Scott, J. E."Identification of non‑oncology drugs with activity against inflammatory and triple‑negative breast cancer cell lines". Oncology Letters 31.2 (2026): 75.
Chicago
Ahmad, S., Kandil, S., Williams, K. P., Scott, J. E."Identification of non‑oncology drugs with activity against inflammatory and triple‑negative breast cancer cell lines". Oncology Letters 31, no. 2 (2026): 75. https://doi.org/10.3892/ol.2025.15428
Copy and paste a formatted citation
x
Spandidos Publications style
Ahmad S, Kandil S, Williams KP and Scott JE: Identification of non‑oncology drugs with activity against inflammatory and triple‑negative breast cancer cell lines. Oncol Lett 31: 75, 2026.
APA
Ahmad, S., Kandil, S., Williams, K.P., & Scott, J.E. (2026). Identification of non‑oncology drugs with activity against inflammatory and triple‑negative breast cancer cell lines. Oncology Letters, 31, 75. https://doi.org/10.3892/ol.2025.15428
MLA
Ahmad, S., Kandil, S., Williams, K. P., Scott, J. E."Identification of non‑oncology drugs with activity against inflammatory and triple‑negative breast cancer cell lines". Oncology Letters 31.2 (2026): 75.
Chicago
Ahmad, S., Kandil, S., Williams, K. P., Scott, J. E."Identification of non‑oncology drugs with activity against inflammatory and triple‑negative breast cancer cell lines". Oncology Letters 31, no. 2 (2026): 75. https://doi.org/10.3892/ol.2025.15428
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