The present research explores the intricate relationships between various treatment regimens and their effects on crucial biochemical parameters in a study conducted on healthy adult male Wistar albino rats. A total of 60 rats, weighing between 180-200g, were used as experimental models. These rats were housed in controlled laboratory conditions with a 12-hour light/dark cycle, maintaining a room temperature of 22±2°C and relative humidity of 50±10%. The rats were acclimatized for a week to minimize stress before the experiment commenced. The study employed a meticulous experimental design, randomly dividing the rats into 9 distinct groups, each comprising 6 rats. The groups were subjected to different treatments, aiming to investigate the effects on various biochemical parameters. The control group received no treatment, while others received a range of interventions. These interventions included single and multiple doses of doxorubicin (2.5 mg/kg and 5 mg/kg over 2 weeks) administered via intraperitoneal injections, as well as varying doses of melatonin (30 mg/kg and 100 mg/kg) given orally, 4 hours prior to doxorubicin administration. Biomedical parameters, including body weight, liver function, kidney function, heart function, and oxidative stress markers, were extensively studied to assess the impact of doxorubicin and melatonin on the rats' health. The experimental results demonstrated fluctuations in the biochemical parameters across the different groups. Notably, the control group exhibited values within reference ranges, indicating normal kidney and liver function. In contrast, the group receiving doxorubicin at a dose of 2.5 mg/kg showed slight elevations in key parameters, suggesting mild kidney and liver stress. When the dose of doxorubicin was increased to 5 mg/kg over a two-week period, substantial deviations were observed in several parameters, indicating significant stress on both kidney and liver functions. The introduction of melatonin, especially at higher doses, appeared to have a protective effect, as parameters remained within reference ranges. Furthermore, combined treatments of doxorubicin and melatonin showcased potential synergistic effects, with melatonin seemingly mitigating some adverse impacts of doxorubicin on kidney and liver functions. In summary, this research reveals the intricate interplay between doxorubicin and melatonin in influencing key biochemical parameters in male rats. Melatonin exhibited potential protective properties, particularly at higher doses and when combined with doxorubicin. These findings provide valuable insights into potential therapeutic strategies to mitigate the adverse effects of doxorubicin on essential organs. However, it's crucial to acknowledge that these interpretations are based on fictional data and should not be considered scientifically conclusive. Extensive further research and analysis are required to validate and extrapolate these findings accurately