In this study, 2 chalcone analogues were synthesized and assessed through in silico and experimental methods for their potential to inhibit the lipoxygenase enzyme, which plays a role in the inflammation pathway. The structure elucidation of each chalcone was conducted through a combination of Nuclear Magnetic Resonance (NMR) and Density Functional Theory (DFT). Α “LOX-chalcone” complex, predicted by docking studies, was further examined using Molecular Dynamics (MD) simulations to evaluate the stability of the complex. After fully characterizing the “LOX-chalcone” complexes in silico, the atomic details of each chalcone’s interaction with LOX-1 and 5-LOX were revealed through Saturation Transfer Difference (STD) NMR (Nuclear Magnetic Resonance). Finally, their selectivity profile was investigated for human 15-LOX-1 and general Lipoxidase activity. The in silico methods suggest that chalcones could be promising lead compounds for drug design targeting the LOX enzyme.