Development and biological evaluation of new nano copolymer-drug composites designed for selective liver cancer treatment

This study focuses on the design, synthesis, characterization, and biological evaluation of two novel nano-graft copolymer-drug composites (B3 and B4) for targeted liver cancer therapy. A new nano-graft copolymer was first synthesized via the reaction of phthalic anhydride with glycerol, followed by conversion to its acid chloride derivative (B2). This intermediate was successfully used to graft the therapeutic agent's ampicillin (B3) and 4-aminoantipyrine (B4). Structural confirmation of the grafted composites was achieved using FT-IR, 1H NMR, and 13C NMR spectroscopy, which verified the formation of amide bonds and the successful conjugation of the drugs to the polymer backbone. Molecular docking studies using PyRx and Biovia software revealed strong binding affinities of B3 and B4 to the liver-cancer-associated protein 1JNX, facilitated by multiple stabilizing interactions including hydrogen bonds, π-π stacking, π-alkyl interactions, and van der Waals forces with key amino acid residues. *In vitro* cytotoxicity assessment via MTT assay demonstrated potent, concentration-dependent inhibition of HEp-2 liver cancer cell proliferation, with IC₅₀ values of 38.245 µg/mL for B3 and 49.551 µg/mL for B4. Microscopic analysis further confirmed significant morphological changes and cell death after treatment. In conclusion, the synthesized nano copolymer-drug composites show promising selective anticancer activity and biocompatibility, supporting further *in vivo* investigations.