The development of new and effective antitumor agents is one of the main goals of medicinal and biochemical research at present. The present study is concerned with the evaluation of the previously synthesized N-Butylpyridoquinoxaline 1,4-dioxide (NBPQD) as an antitumor agent against Ehrlich ascites carcinoma (EAC). The first part of the study (tumor imaging) was to investigate the biodistribution of NBPQD in the organs of EAC-bearing mice using iodine-125 isotope stressing on its distribution in the main organs (stomach, liver, spleen, kidney) in addition to blood and ascetic fluid. The second part was the assessment of the antitumor activity of NBPQD by estimating the tumor volume and the contents of total protein, total lipid, DNA and RNA in liver tissues. In addition, liver function tests and the redox status were assessed. Tumor volume and DNA, RNA, urea and malondialdehyde (MDA) levels and the liver enzymes activity were highly significantly increased (P < 0.001) in untreated EAC-bearing mice compared to control. However, total lipid and total protein in liver tissues in addition to serum albumin, glucose, reduced glutathione (GSH) as well as activities of glutathione reductase (GSH-R) and superoxide dismutase (SOD) all were highly significantly decreased in untreated EAC-bearing mice compared to controls. All these decreased parameters were highly significantly restored to their normal levels in NBPQD treated mice compared to the untreated EAC-bearing mice. The survival time of the NBPQD treated mice was longer than that of the untreated ones. It is thus, evident that NBPQD had a remarkable antitumor activity against EAC in Swiss albino mice.