Inflammation is a central component of numerous pathological conditions, and the limitations of currently available anti-inflammatory therapies continue to motivate the search for structurally diverse therapeutic candidates. Mitragyna speciosa Korth. contains bioactive indole alkaloids, particularly mitragynine and 7-hydroxymitragynine, that have been associated with anti-inflammatory effects. This study evaluated the predicted binding affinity and ligand-protein interaction profiles of mitragynine and 7-hydroxymitragynine against four inflammation-related targets—cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), interleukin-1 beta (IL-1β), and tumor necrosis factor-alpha (TNF-α)—using molecular docking. Protein structures were obtained from the Protein Data Bank (PDB IDs 1EQG, 3LN1, 5R87, and 3EWJ, respectively). Protein and ligand preparation and interaction visualization were performed using Discovery Studio Visualizer, docking was conducted using PLANTS, and protocol validation was assessed by redocking and root mean square deviation (RMSD) analysis in Molegro Molecular Viewer. All protocols met the prespecified RMSD acceptance criterion of ≤2.0 Å, with values of 1.57 Å for COX-1, 0.77 Å for COX-2, 1.36 Å for IL-1β, and 0.55 Å for TNF-α. Within each protein target, 7-hydroxymitragynine showed the most favorable predicted score among the tested alkaloids for COX-1 and COX-2 (−17.39 and −19.56 kcal/mol, respectively), whereas mitragynine showed the more favorable predicted score for IL-1β and TNF-α (−16.72 and −20.42 kcal/mol, respectively). These results indicate target-dependent binding behavior and provide a computational basis for further structural, pharmacokinetic, and experimental evaluation of both alkaloids as potential anti-inflammatory candidates.