In the present study, a series of cardo-type novel polyimides (PI)s containing xanthene groups was synthesized. All the polymers were amorphous in nature with inherent viscosities in the range of 0.72–0.91 dl/g. Study of thermal behavior showed that PIs have excellent thermal stability with 10% weight loss at temperatures (Td10%) between 426 and 528 C, glass transition-related temperatures (Tg) in the range of 292–305 C, and weight residual at 700 C ranged from 55% to 68% under N2 atmosphere. Xanthene and fluorene presence, progressed the PIs solubility and also induced good biological activities. The in vitro anticancer activity of the 6FD-derived PI against the A431 (epidermoid carcinoma) cells showed an IC50 value of 29.3 μM. In the next part of the work, a polymer-based composite (PI-B/m-TiO2@HBP) was prepared by inscribing 20 wt% of hyperbranched polyamide-grafted TiO2 nanoparticles into BTDA-derived PI-B matrix, and compared with the unfilled PI, its photoluminescence intensity and thermal stability properties were increased. The PI-B and nanocomposite exhibited good antibacterial activity against six bacterial strains. In the following, study of methylene blue (MB) dye adsorption from aqueous solution by the obtained PIs and PI-B/HBP@TiO2 was carried out and removal efficiency values were found above 80.62%