After preparing a series of new dispersed dyes based on enaminone derivatives, verifying the validity of the chemical compositions using the latest modern scientific methods, studying them, and publishing the results that indicate the effectiveness of these new dispersed dyes against a group of bacteria and cancer cells. In this study we evaluated the cytotoxic effects of the synthesized dyes using the normal cell line, lung WI-38, where the dyes showed satisfactory results and are promising towards their safe use on human lung fibroblast normal cells WI-38. As anti-proliferative agents with good selectivity index range (1.65–6.42), the dyes 5a–5f under investigation shown non-significant cytotoxic impact with CC50 values of 7.71, 24.49, 21.01, 104.51, 33.74, and 115.67 mM, respectively. These results provided a favourable safety profile.