Skrining Virtual In Silico dan Uji In Vitro Senyawa Inhibitor Multi Target CHK1, CHK2, dan WEE1 yang Berpotensi sebagai Kandidat Obat Anti Kanker

Abstract

Breast cancer and ovarian cancer are cancers with the highest prevalence and mortality rates in women worldwide. CHK1, CHK2, and WEE1 are molecular pathways involved in many cancer cases. Inhibition of CHK1, CHK2, and WEE1 as checkpoints can trigger apoptosis because cells with damaged DNA will enter mitosis without repair. Prexasertib is a selective inhibitor of CHK1 and CHK2, while adavosertib is a selective inhibitor of WEE1. Both inhibitors have undergone clinical trials but have side effects such as neutropenia, anemia, and hepatotoxicity. The objective of this research is to find natural compound candidates that have the potential to act as multi-target inhibitors against CHK1, CHK2, and WEE1 with minimal side effects. Potential compounds were identified through pharmacophore modeling, followed by molecular docking and molecular dynamics. The in silico test results were then analyzed and followed by in vitro tests to assess cytotoxic activity, proliferation, cell cycle, apoptosis, and protein expression in 4T1 and MCF-7 cells. The study identified six potential compounds from molecular docking and molecular dynamics simulations. Based on interaction analysis, compounds MCULE-6508263597-0, MCULE-1328845163-0, MCULE-3015027877-0, and MCULE-6752415685-0 were selected for in vitro testing on 4T1 and MCF-7 cells. Compounds MCULE-3015027877-0 and MCULE-6752415685-0 showed better IC50 values compared to the controls, with decreased cell viability over 72 hours. These two compounds showed activity in the S and G2M phases in both cell lines and, in apoptosis tests, enhanced early and late apoptosis activity in both cell lines. In protein expression tests, both compounds informed lower parent cell percentages compared to controls and reference compounds, which indicated the inhibitory activity of CHK1, CHK2, and WEE1. The results of this study conclude that two compounds, identified through pharmacophore modeling, molecular docking, molecular dynamics, and in vitro testing, have activity as multi-target inhibitors against CHK1, CHK2, and WEE1.