Synthesis of ethyl 2-((3-(trifluoromethyl)phenyl)amino)benzoate, (Ester) (Mishra et al. 2008)

Synthesis of 2-((3-(trifluoromethyl)phenyl)amino)benzohydrazide, (hydrazide) (Abbas et al. 2015; Hmood et al. 2021)

General method for synthesis of the compounds (1–5) (Al-Saad et al. 2019)

N-(4-chlorophenyl)-2-(2-((3-(trifluoromethyl)phenyl)amino)benzoyl)hydrazine-1-carbothioamide (1)

N-(4-fluorophenyl)-2-(2-((3-(trifluoromethyl)phenyl)amino)benzoyl)hydrazine-1-carbothioamide (2)

N-(4-bromophenyl)-2-(2-((3(trifluoromethyl)phenyl)amino)benzoyl)hydrazine-1-carbothioamide (3)

N-phenyl-2-(2-((3-(trifluoromethyl)phenyl)amino)benzoyl)hydrazine-1-carbothioamide (4)

N-(p-tolyl)-2-(2-((3-(trifluoromethyl)phenyl)amino)benzoyl)hydrazine-1-carbothioamide (5)

General method for synthesis of the compounds (6–10) (Szeliga and Monika 2020)

N-(4-chlorophenyl)-5-(2-((3-(trifluoromethyl)phenyl)amino)phenyl)-1,3,4-thiadiazol-2-amine (6)

N-(4-fluorophenyl)-5-(2-((3-(trifluoromethyl)phenyl)amino)phenyl)-1,3,4-thiadiazol-2-amine (7)

N-(4-bromophenyl)-5-(2-((3-(trifluoromethyl)phenyl)amino)phenyl)-1,3,4-thiadiazol-2-amine (8)

N-phenyl-5-(2-((3-(trifluoromethyl)phenyl)amino)phenyl)-1,3,4-thiadiazol-2-amine (9)

N-(p-tolyl)-5-(2-((3-(trifluoromethyl)phenyl)amino)phenyl)-1,3,4-thiadiazol-2-amine (10)

Cell culture

In vitro cytotoxicity (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) MTT assay

Detection of apoptosis

Quantitative reverse transcription-polymerase chain reaction (qRT-PCR)

Western blot analysis

Cytochrome c release apoptosis assay

EGFR kinase inhibitory assay

Statistical analysis

Cytotoxicity of the target derivatives is cancer-cell specific

Cytotoxicity of compound 7 is attributed to induction of apoptosis

Compound 7 induction of apoptosis is dependent on activation of caspase 9

Compound 7 exhibit EGFR kinase inhibitory activity

In-silico ADMET, and carcinogenicity analysis