Synthesis of ethyl 2-((3-(trifluoromethyl) phenyl) amino) nicotinate (1) (Al-Bayati et al. 2021)

Synthesis of 2-((3-(trifluoromethyl) phenyl) amino) nicotinohydrazide (2) (Hmood et al. 2021)

General method for synthesis of niflumic hydrazine- carboamide derivatives (3A, 3B, and 3C) (Nederlof 1963)

General method for synthesis of niflumic hydrazine- carbothioamide derivatives (4A, 4B, 4C, and 4D) ()

General method for synthesis of niflumic1,2,4-triazole-3-thione derivatives (5A,5B, and 5C) (Abdul- Jabbar et al. 2005)

Synthesis of 4-(4-chlorophenyl)-5-(2-((3-(trifluoromethyl) phenyl) amino) pyridin-3-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (6) (Abdul- Jabbar et al. 2005)

Cell culture

Analysis of cytotoxicity by the in vitro MTT assay

Cell cycle analysis by flow cytometry

Detection of viable, apoptotic, and necrotic cells by flow cytometry

Analysis of apoptosis markers gene expression by quantitative reverse transcription-polymerase chain reaction (qRT-PCR)

Vascular endothelial growth factor receptor 2 (VEGFR2) and epidermal growth factor receptor (EGFR) kinase inhibitory assay

ADMET studies

Density Functional Theory (DFT)

Molecular dynamics (MD) simulations

The effects of the tested compounds on Hep G2 and A549 cancer cell proliferation

Compound 4C-treated Hep G2 hepatocyte carcinoma cells exhibit disruption in cell cycle progression

The cytotoxicity of compound 4C against Hep G2 hepatocyte carcinoma cells is attributed to apoptosis

Compound 4C targets VEGFR-2 kinase and compound 5B targets EGFR kinase

3D-QSAR model generation and validation

3D-QSAR ValidationVEGFR-2 TK

3D QSAR Validation EGFR TK