Evaluation of 2-Thioxoimadazolidin-4-one Derivatives as Potent Anti-Cancer Agents through Apoptosis Induction and Antioxidant Activation: In Vitro and In Vivo Approaches

Abstract

Background: Hepatocellular carcinoma (HCC) is one of the most widespread malignancies and is reported as the fourth most prevalent cause of cancer deaths worldwide. Therefore, we aimed to investigate the probable mechanistic cytotoxic effect of the promising 2-thioxoimidazolidin-4-one derivative on liver cancer cells using in vitro and in vivo approaches. The compounds were tested for the in vitro cytotoxic activity using MTT assay, and the promising compound was tested in colony forming unit assay, flow cytometric analysis, RT-PCR, Western blotting, in vivo using SEC-carcinoma and in silico to highlight the virtual mechanism of action. Both compounds 4 and 2 performed cytotoxic effects against HepG2 cells with IC50 values of 0.017 and 0.18 μM, respectively, compared to Staurosporine and 5-Fu as reference drugs with IC50 values of 5.07 and 5.18 µM, respectively. Compound 4 treatment revealed apoptosis induction by 19.35-fold (11.42% compared to 0.59% in control), arresting the cell cycle at G2/M phase. Moreover, studying gene expression that plays critical roles in cell cycle and apoptosis by RT-PCR demonstrated that compound 4 enhances the expression of the pro-apoptotic genes p53, PUMA, and Caspase 3, 8, and 9, and impedes the anti-apoptotic Bcl-2 gene in the HepG2 cells. It can also inhibit the PI3K/AKT pathway at both gene and protein levels, which was reinforced by the in silico predictions of the molecular docking simulations towards the PI3K/AKT proteins. Finally, in vivo study verified that compound 4 has a promising anti-cancer activity through activating antioxidant levels (CAT, SOD and GSH) and ameliorating hematological, biochemical, and histopathological findings.

Synthesis, X-ray Single-Crystal Analysis, and Anticancer Activity Evaluation of New Alkylsulfanyl-Pyridazino[4,5-b]indole Compounds as Multitarget Inhibitors of EGFR and Its Downstream PI3K-AKT Pathway

“…Figure 5 shows that the compound treatment increased the cell population by 46.32% in pre-G phase compared to 2.19% in the control and by 46.12% in S phase compared to 33.61% in the control, while the cell population was not significantly decreased. These findings are consistent with previous studies [38], which evaluated apoptosis-inducing activity using flow cytometric apoptosis and cell cycle analyses.…”

Effect of 2-Thioxo Imidazolidin-4-Ones derivative copper complex on the oxidative state on the male rats exposed to hydrogen peroxide

“…With treatment of copper complex of 2-Thioxo Imidazolidin-4-Ones as antioxidants, antioxidant levels were significantly increased to (0.074) and it was observed that GSH was increased to (0.075), Catalase was increased to (0.068), These results are in accordance with (Nafie et al,2021) antioxidant levels were significantly increased, Because the treatment caused the regulation of apoptosis.…”

Development of Novel 1,3-Disubstituted-2-Thiohydantoin Analogues with Potent Anti-Inflammatory Activity; In Vitro and In Silico Assessments

“…The discovery of new effective anti-inflammatory agents, thus, considered as an urgent concern [ 4 , 5 ]. Several reports demonstrated 2-thioxoimidazolidine-4-one analogues as a potent class of compounds which possess a broad-spectra of pharmacological and biological activities [ 64 , 65 , 66 ]. Among synthesized and evaluated compounds, compound 5 and 7 exhibited the most potent cytotoxic activity against LPS-activated RAW264.7 cells (IC 50 of 212.3 μg/mL and 197.68 μg/mL, respectively), comparable to that of celecoxib drug (IC 50 value 251.2 μg/mL) ( Figure 4 ).…”

“…The cycling conditions were initial heating stage 5 min at 95 °C, then 45 cycles of denaturation phase at 95 °C 10 s, annealing phase at 60 °C 30 s, and extension phase at 72 °C 1 min using iScript TM One-Step RT-PCR Kit with SYBR ® Green (Bio-Rad, Hercules, CA, USA). Finally, the expression of relative gene was carried out using 2 −ΔΔct equation and normalized to β-actin gene [ 66 , 97 ]. Primers of IL-6 were F 5′-AGACAGCCACTCACCTCTTCAG-3′, R 5′-TTCTGCCAGTGCCTCTTTGCTG-3′; primers of TNF-α were F 5′-CTCTTCTGCCTGCTGCACTTTG-3′, R 5′-ATGGGCTACAGGCTTGTCACTC-3′; primers of β-actin were F 5′-GCACCACACCTTCTACAATG-3′, R 5′-TGCTTGCTGATCCACATCTG-3′.…”