Nattanan Jiwacharoenchai [a], Duangnapa Kiriwan [a], Lueacha Tabtimmai [b], Supaphorn Seetaha [c], Sirilata Yotphan [d], Chonticha Suwattanasophon [e] and Kiattawee Choowongkomon [a,b,c,f]*
                  [a] Genetic Engineering Interdisciplinary Program, Graduate School, Kasetsart University, Bangkok, 10900 Thailand
                  [b] Department of Biotechnology, Faculty of Applied Science, King Mongkut University of North Bangkok, Bangkok, 10800 Thailand
                  [c] Center for Advanced Studies in Nanotechnology for Chemical, Food and Agricultural Industries, KU Institute for Advanced Studies, Kasetsart University, Bangkok, 10900 Thailand
                  [d] Department of Chemistry, Faculty of Science, Mahidol University, Bangkok, 10400 Thailand
                  [e] Department of Physiological Chemistry, Faculty of Chemistry, University of Vienna, Vienna, 1010, Austria
                  [f] Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok, 10900 Thailand *Author for correspondence; e-mail: kiattawee.c@ku.th
              EGFR plays an extensive role in the signaling pathway such as proliferation, migration, invasion. However, the malfunctioning of the epidermal growth factor receptor (EGFR) is a crucial factor in cancer progression. The search for new kinase inhibitors is necessary due to the evolving nature of diseases and the emergence of resistant strains, prompting the need for novel compounds that can effectively target and inhibit specific kinases for therapeutic purposes. This study explores quinoxalinone derivatives, comprised of benzene and pyrazine aromatic rings, as novel potential EGFR kinase inhibitors for cancer therapy, fulfilling an essential need in targeted cancer treatments. We utilized molecular docking to examine 46 synthesized quinoxalinone derivatives for their interaction with the tyrosine kinase domain of EGFR. Fifteen compounds exhibited strong binding affinities. Notably, LS3c, MN333, and MN343 displayed significant inhibitory effects on A431 cells with half-maximum inhibitory concentration (IC50) values comparable to gefitinib, erlotinib, and afatinib in enzymatic kinase assays. In particular, MN343, the most potent compound, was selected for molecular dynamic simulation to understand its interaction with the tyrosine kinase of EGFR. Further validation through additional experiments and animal studies, including a clinical trial, is warranted.