Validation of Novel Molecule as BRAF V600E Inhibitor for the Treatment of Colorectal Cancer

Aim: To validate 3-(6,7-dimethoxy-3,4-dihydroisoquinoline-2-carbonyl)-N-(2-methoxyphenyl)benzenesulphonamide (B1) as a potential BRAF V600E inhibitor via in vitro analyses.

Study Design: Tests were carried out on HT-29 human colon cancer cell lines. Various studies such as clonogenic assay, apoptotic studies, Reactive Oxygen Species (ROS) studies and Indirect ELISA tests were carried out in both the control sample and B1-incorporated sample for determining the efficacy of B1.

Place and Duration of Study: The study was conducted at Sahrdaya College of Engineering and Technology, Kodakara and Centre for Research on Molecular and Applied Sciences, Thiruvananthapuram, between October 2023 and February 2024.

Methodology: In this study, many in vitro analyses were carried out to verify the BRAF V600E inhibition ability of B1 to be used as a potential drug candidate against the treatment of CRC. HT-29 human colon cancer cell lines were grown. Following that, clonogenic assay was performed to measure the cell growth, apoptotic studies to figure out the cell vitality, Reactive Oxygen Species (ROS) studies to calculate the amount of ROSs present, and Indirect ELISA tests to quantify the total protein concentration were performed in both control sample and B1-incorporated sample.

Results: The molecule B1 significantly reduced the colony formation, increased the cell mortality, and elevated ROSs levels. The concentration of ERK (the downstream protein of BRAF) was likewise significantly reduced in the B1 sample, implying that the substance B1 is an efficient inhibitor of BRAF. All the results pointed to the fact that B1 can be used as an effective BRAF V600E inhibitor.

Conclusion: This work established the effectiveness of the molecule B1 as an effective inhibitor against BRAF for the treatment of CRC.