The present study was carried out to utilize different levels of germinated pumpkin seed flour (GPSF), as a source of protein and minerals in fortification of wheat flour for making pan bread. Organoleptic properties of prepared pan bread and its biological effect on lipids profile of rats fed on hypercholesterolemic diet were studied. Pan bread was prepared with GPSF at different levels. The results showed a significant decrease in fat content and an increase in protein, fiber contents and caloric value by increasing defatted GPSF replacement level. Biological assay indicated that food intake, protein intake, feed efficiency ratio and protein efficiency ratio of rats were not significantly affected by defatted GPSF replacement. Serum TC, TG and LDL-C were significantly decreased by replacement with 20% defatted GPSF while HDL-C increased and caused a beneficial effect in improving the lipids profile of rats fed on hypercholesterolemic diet.