Aims: The present study was conducted to examine the inhibitory effects of synthesized
sulfonylhydrazones on the expression of CD73 (ecto-5′-NT).
Background: CD73 (ecto-5′-NT) represents the most significant class of ecto-nucleotidases, which
are mainly responsible for the dephosphorylation of adenosine monophosphate to adenosine. Inhibition
of CD73 played an important role in the treatment of cancer, autoimmune disorders, precancerous
syndromes, and some other diseases associated with CD73 activity.
Objective: Keeping in view the significance of CD73 inhibitor in the treatment of cervical cancer, a
series of sulfonylhydrazones (3a-3i) derivatives synthesized from 3-formylchromones were evaluated.
Methods: All sulfonylhydrazones (3a-3i) were evaluated for their inhibitory activity towards CD73
(ecto-5′-NT) by the malachite green assay and their cytotoxic effect was investigated on the HeLa cell
line using MTT assay. Secondly, the most potent compound was selected for cell apoptosis, immunofluorescence
staining, and cell cycle analysis. After that, CD73 mRNA and protein expression were
analyzed by real-time PCR and Western blot.
Results: Among all compounds, 3h, 3e, 3b, and 3c were found to be the most active against rat-ecto-
5′-NT (CD73) enzyme with IC50 (μM) values of 0.70 ± 0.06 μM, 0.87 ± 0.05 μM, 0.39 ± 0.02 μM,
and 0.33 ± 0.03 μM, respectively. These derivatives were further evaluated for their cytotoxic potential
against cancer cell line (HeLa). Compounds 3h and 3c showed the cytotoxicity at IC50 value of
30.20 ± 3.11 μM and 86.02 ± 7.11 μM, respectively. Furthermore, compound 3h was selected for cell
apoptosis, immunofluorescence staining, and cell cycle analysis, which showed a promising apoptotic
effect in HeLa cells. Additionally, compound 3h was further investigated for its effect on the expression
of CD73 using qRT-PCR and western blot.
Conclusion: Among all synthesized compounds (3a-3i), Compound 3h (E)-N'-((6-ethyl-4-oxo-4Hchromen-
3-yl) methylene)-4-methylbenzenesulfonohydrazide was identified as the most potent compound.
Additional expression studies conducted on the HeLa cell line proved that this compound
successfully decreased the expression level of CD73 and thus, inhibited the growth and proliferation
of cancer cells.