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Background: Sono-photodynamic therapy (SPDT) is a novel cancer treatment therapy that uses the light of a particular wavelength and sound of a particular frequency to activate a light- and sonosensitive material that attaches selectively tumor cells, causing their breakdown. The present work aims to investigate biochemically and pathologically the effectiveness of using SPDT in combination with either nano-5-aminolevulinic acid (nano-ALA) or nano-Chlorophyll (nano-Chl), as sono-photosensitizers, in curing Ehrlich ascites carcinoma tumor implanted to a group of mice. Methods: A total of 130 male Swiss albino mice age 60–65 days, weighing 20 ± 2.0 g, were used in this experiment. Two sources of energy were used; namely infrared laser (IR) and Ultrasound for 3 min. When the tumor had grown to about 10 mm in diameter at day 10 after inoculation, the treatment strategy was started. Six experimental groups were investigated. Biochemical analyses were carried out to detect the serum levels for activities of oxidative stress enzymes, liver, and kidney functions. Pathological analyses were carried out to evaluate the effect of different treatment modalities on tumor treatment. Results: It was observed that SPDT utilizing nano-ALA or nano-Chl ameliorated the activity levels of the studied enzymes in mice bearing tumor lesions, indicating the safe and non-toxic treatment modality. The pathological evaluation showed that targeted SPDT caused the almost complete destruction of cancer cells. Conclusion: The results suggest that nano-ALA or nano-Chl could be used as a novel and safe nano-materials with great potential as an effective drug delivery system in cancer-targeted SPDT.