Prostate cancer is a prevalent and highly heterogeneous malignancy that affects men globally. Despite the availability of various treatment targets, Cytochrome P450 (CYP) enzymes have gained significant attention due to their crucial role in metabolizing both endogenous and exogenous compounds. This study explores Diosmetin as a potential CYP antagonist for treating prostate cancer. To evaluate Diosmetin's potential as a CYP antagonist, we employed a comprehensive in silico approach. Molecular docking was conducted using the Glide software to assess the binding affinity of Diosmetin with CYP enzymes, specifically CYP17A1 and CYP19A1, which are associated with prostate cancer. The druglike properties of Diosmetin were evaluated, focusing on its pharmacokinetic attributes. Additionally, Diosmetin's ADMET (Absorption, Distribution, Metabolism, Excretion, and Toxicity) characteristics were analyzed to determine its suitability as a therapeutic agent. Molecular dynamics (MD) simulations were performed using Desmond to assess the stability and persistence of Diosmetin binding with the CYP enzymes over a 200 ns simulation period. Molecular docking studies revealed robust binding affinities between Diosmetin and CYP17A1 (- 11.261 kcal/mol) and CYP19A1 (- 11.145 kcal/mol). Diosmetin demonstrated favorable pharmacokinetic properties and advantageous ADMET characteristics, including high bioavailability, good dispersion, and favorable metabolism. MD simulations indicated persistent binding interactions between Diosmetin and the CYP enzymes throughout the 200 ns simulation, reinforcing the reliability of these interactions. Pharmacoinformatics investigations provide valuable insights into the potential of Diosmetin as a promising lead compound for the development of novel drug candidates against prostate cancer. The strong binding affinity and favorable pharmacokinetic and ADMET profiles suggest that Diosmetin could be an effective CYP antagonist and warrants further investigation as a potential therapeutic agent for prostate cancer.