A novel inhibitor, 2-cyano-3-(1-phenyl-3-(thiophen-2-yl)-pyrazol-4-yl)acrylamide linked to sulphamethoxazole, blocks anti-apoptotic proteins via molecular docking and strongly induced apoptosis of HCT116 cell line by different molecular tools

Abstract

A new series of cyanoacrylamides incorporating sulphamethoxazol were prepared and confirmed by different spectral tools. Anticancer screening of the new compounds was done against three different types of carcinoma cell lines involving (A549, HCT116, and MDA) using MTT assay. Compound 7 among all tested derivatives achieved the best cytotoxic effect against all tested carcinoma cell lines. HCT116 revealed the best sensitivity and cytotoxic activity toward compound 7 relative to 5-FU. The target compound offered less toxic effect when tested on normal melanocytes (HFB4). Simulation modeling studies revealed strong binding affinity toward the following domains (1dls, 2c6o, and 2wgj) and moderate binding modes toward (3eyl, 4kmp, 2w3l, and 5lab) domains with different binding energy scores. Gene expression profile outlined that caspase-3, BAX, and P53 genes were strongly upregulated relative to their control, while BCL2, MMP1, and CDK2 were effectively down regulated assuming the activation of the apoptotic pathway. Flow cytometry technique revealed that compound 7 stimulated cell cycle arrests at the G2/M phase. Other extensive molecular diagnostic tools were utilized in this report as ELISA, DPA, SEM, and TEM assays which confirmed that our target novel compound 7 was a very promising and interesting chemotherapeutic agent with less toxic effect. Also, authors herein suggested that additional sulphamethoxazole linked to 3-(1-phenyl-3-(thiophen-2-yl)-1H-pyrazol-4-yl)acrylonitrile in compound 7 was responsible for its promising cytotoxic activity against colorectal carcinoma cell line.

Keywords

• Colorectal carcinoma
• Cyanoacrylamides
• Sulphamethoxazole
• Cytotoxicity
• Modeling studies
• Apoptosis