Male prostate cancer (PCa) is considered among the most fatal illnesses. Despite the recent decrease in prostate cancer incidence attributed to advancements in early detection and therapy, these reductions have not effectively mitigated the elevated fatality rate linked to this disease. The drug Nilutamide was effectively radiolabeled with technetium-99m, producing a radiochemical yield of 96 ± 0.14 % under optimal conditions. In our study, two cohorts of mice were utilized, namely the control group and the group with prostate cancer. Various biochemical parameters, including PSA levels in serum, were assessed, revealing a significantly elevated value in the group with prostate cancer, indicating potential tumor development. Furthermore, the activities of antioxidant enzymes (CAT, SOD) were notably lower in the group with prostate cancer compared to the healthy control group, while the oxidative activity reflected by MDA levels, the final product of lipid peroxidation, was higher in the prostate cancer group than in the healthy control group. The biodistribution analysis showed rapid localization of 99mTc-Nilutamide in prostate cancer tissue after 2 h post-injection, with a substantial value of 11.4 ± 1.1 % I. D/g tissue. Consequently, it was deduced that radiolabeled 99mTc-Nilutamide can serve as an effective imaging tool for prostate cancer.