Canceris uncontrolled cellular growth of abnormal cells.This abnormal growth result from interaction of genetic factors andexternal riskfactors. It considers as animportantcauseofmortality andmorbidity worldwide, in 2020 it was found that there were 19.3 million new cases of cancer and near to 10.0 million cancer-related deaths worldwide. This research aimed to prepare benzamide derivatives and to evaluate their in-vitro cytotoxic activity using SRB (Sulforhodamine B) assay test against two cell lines including human colorectal carcinoma (CaCo-2) cell line and breast adenocarcinoma (MCF-7) cell line. These compounds were prepared adopting the direct coupling of the 2-methylaminobenzamide with the acyl chlorides in anhydrous tetrahydrofuran (THF) without catalyst. The majority of the compounds were produced with a good yield, and their structures were confirmed by nuclear magnetic resonance (NMR), Fourier transform infrared spectroscopy (FTIR),Gas chromatography-mass spectroscopy (GC-MS) and high resolution mass spectroscopy (HRMS). The compounds were then evaluated in vitro using the SRB assay test, which involved three trials for each compound 3, 4, and 5 with Doxorubicin (as a reference drug).Compound 5has potentcytotoxic activity against human colorectal carcinoma (CaCo-2) cell line and breast adenocarcinoma (MCF-7) cell line, with a IC50 values of 7.13 ?M and 6.82 ?M, respectively, in the tested concentration range (0.01-100) ?M. Compounds 3 and 4 have no cytotoxic effect against human colorectal carcinoma (CaCo-2) and breast adenocarcinoma (MCF-7) cell lines.Our findings showed that compound 5 might serve as a fundamental scaffold for future structural design in the production of potent anticancer drugs.