Background: Sono-photodynamic therapy (SPDT) is a safe, non-toxic and non-invasive way of destroying cancer cells, as well as enhancing immune protective function. The present work aims to cure Ehrlich ascites carcinoma tumor implanted in a group of mice using a sonophotodynamic modality in combination with nano-chlorophyll (nano-Chl) as a sonophoto sensitizer. 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 study was started. Six experimental groups were investigated. The dimensions of the tumor, tumor volume, tumor mass inhibition ratio, and tumor volume growth ratio were calculated. Results: The results revealed that the effect of exposing the tumor to IR laser alone or in the presence of the nano- Chl resulted in decreasing the tumor volume, tumor growth rate, and inhibition ratio. A similar result was observed in the case of using ultrasound exposure only or in the presence of the nano-Chl.  Combined treatment of IR laser at 7000 Hz and pulsed ultrasound wave in the presence of nano-Chl was more effective than either IR laser or ultrasound alone. Conclusion: It can be suggested that folic acid-conjugated nano-Chl sono-photosensitizers (FA-NGO-Chl) could be used as a novel nanomaterial with great potential as an effective drug delivery system in targeting sono-photodynamic-therapy (SPDT).