Introduction: Cyclooxygenases (COXs) are the enzymes that specifically influence the production (minute quantities) of prostaglandins and their derivatives, which are responsible for discomfort, inflammation, and other symptoms. The study aimed to develop certain new phenoxyacetanilide derivatives in multi-step synthesis and to screen their anti-inflammatory and analgesic perspectives along with docking studies against main inflammatory target COX-2 enzyme as well as enzyme-linked immunosorbent assay (ELISA) analysis against two prominent inflammatory mediators; Interleukin-6 (IL-6) and COX-2 enzyme. Methods: The newly synthesized substances were developed through multi-step schemes and were analyzed thoroughly employing advanced analytical techniques such as Fourier-Transformed Infrared Spectroscopy (FT-IR), Proton-Nuclear Magnetic Resonance Spectroscopy (1 H-NMR), Mass spectroscopy, and Elemental Analysis. The possibility of a binding site and binding strength of new acetanilide derivatives were discovered using a molecular docking study against COX-2 enzyme using AutoDock Vina software. All of the synthesized derivatives were tested for analgesic activity (using Eddy__ampersandsignrsquo;s hot plate method) and anti-inflammatory effects (employing the carrageenan-induced paw edema method). Results: As compared to the standard drug diclofenac sodium, (2-[2-methoxy-4-(prop-2-en-1-yl)phenoxy]-N-(2-methylphenyl) acetamide) (RKS-1) expressed both strong analgesic as well as anti-inflammatory activity along with demonstrating the lowest docking score (-8.9 Kcal/mol) against COX-2 enzyme. Conclusion: RKS-1 was found to be the most potent lead. The in vivo results were found to be successfully correlated with the obtained in silico data. The current study will draw the attention of global chemists towards the rational development of newer synthetic phenoxyacetanilide derivatives with pronounced non-steroidal anti-inflammatory drugs (NSAID) activity.