A Potent EGFR Inhibitor, N-Phenyl Pyrazoline Derivative Suppresses Aggressiveness and Cancer Stem Cell-Like Phenotype of Cervical Cancer Cells

Abstract

Objective Metastasis causes approximately 90% of cancer-related deaths, including in cervical cancer patients. Uncontrolled cell proliferation, migration, and cancer stemness act as critical events in primary tumor growth and cancer metastasis progression in cervical cancer. Here, we investigated the anti-proliferative, anti-migration, and cancer stemness inhibition activity of N-phenyl pyrazoline derivatives against cervical cancer cells. Methods The chalcone and phenylhydrazine were used to synthesize the N-phenyl pyrazoline 2/5 (P2 and P5). The MTT, colony formation, and wound healing assays were performed to evaluate the N-phenyl pyrazoline effect in HeLa cells. The N-phenyl pyrazoline’s protein target was predicted using SwissTargetPrediction and AutoDock Vina software. The Western blotting assay was performed to evaluate the target proteins. The public dataset analysis was used to confirm the clinical relevance of target protein in cervical cancer patients. Results N-phenyl pyrazoline 2 and 5 were successfully synthesized. The N-phenyl pyrazolines 2 and 5 exhibit cytotoxic effect in HeLa cell line with 20.26 µM, 4.708 µM of IC 50, respectively. Further study shows that the N-phenyl pyrazoline 5 suppresses the cell proliferation and migration ability of HeLa cell line in a dose-dependent manner. Target prediction and molecular docking reveal that EGFR and ERBB2 protein as the main target of the N-phenyl pyrazoline 5 compound. The N-phenyl pyrazoline 5 suppresses the EGFR expression level but not the total ERK1/2. Public data and GSEA analysis found that the EGFR high expression level is positively associated with poor survival, cancer metastasis-related signaling pathways, and cancer stem cell markers in cervical cancer patients. In addition, the N-phenyl pyrazoline 5 reduces the HeLa’s tumorsphere size and cancer stem cell marker, CD133. Conclusion N-phenyl pyrazoline 5 suppresses the cell viability, proliferation, migration, and cancer stem cell-like phenotype of cervical cancer cells via EGFR inhibition.

Nitrogen‐Containing Heterocyclic Scaffolds as EGFR Inhibitors: Design Approaches, Molecular Docking, and Structure‐Activity Relationships

“…On the other hand, compound 55 b suppressed the EGFR expression level but not the total ERK1/2. The docking analysis revealed that compound 55 b displays a improved binding affinity to the EGFR than the EGFR inhibitor (erlotinib) [127] …”

“…The docking analysis revealed that compound 55 b displays a improved binding affinity to the EGFR than the EGFR inhibitor (erlotinib). [127] [128] In 2019, George and group reported the pyrazoline clubbed quinoline hybrid and appraised against anti-tumor potential. The in vitro assessment of SRB assay against MCF-7), HeLa and DLDI specified that nearly all the candidate showed noticeable inhibitory effect with IC 50 ranging from 0.064-4.94, 0.12-6.273 and 0.038-3.155 μM respectively.…”

N-phenyl pyrazoline derivative inhibits cell aggressiveness and enhances paclitaxel sensitivity of triple negative breast cancer cells

“…In the previous study, we found a small compound, N -phenyl pyrazoline inhibits one of the protein kinases, EGFR leads to cervical cancer aggressiveness attenuation 6 . N -phenyl pyrazoline also inhibits the cell viability of human breast cancer cells and colorectal cancer cells 7 .…”

Pyrazoline B-Paclitaxel or Doxorubicin Combination Drugs Show Synergistic Activity Against Cancer Cells: In silico Study

“…[13][14][15] This compound also induces poor survival, reduces tumor sphere size, inhibits cell growth, and reduces CD133 levels in HeLa cervical cancer cell lines. 14,16 We successfully showed a decrease in EGFR (Epidermal Growth Factor Receptor) levels in HeLa cells induced by an N-phenyl pyrazoline derivate. 13,16,17 Therefore, the N-phenyl pyrazoline derivative is a promising anticancer drug and potential to be combined with other anticancer drugs to increase their effectiveness.…”

“…14,16 We successfully showed a decrease in EGFR (Epidermal Growth Factor Receptor) levels in HeLa cells induced by an N-phenyl pyrazoline derivate. 13,16,17 Therefore, the N-phenyl pyrazoline derivative is a promising anticancer drug and potential to be combined with other anticancer drugs to increase their effectiveness. We previously reported that a compound belonging of N-phenyl pyrazoline derivates, Pyrazoline B (methoxyphenyl)-1-phenyl-3-(thiophen-2-yl) −4,5-dihydro-1H-pyrazole), exerts promising anticancer activity based on in silico 18 and in vitro 19 studies.…”