Synthesis, characterization and in vitro antibacterial assessments of a novel modified poly[maleic anhydride-alt-acrylic acid]/acriflavine conjugate

Water-soluble maleic anhydride-containing poly[maleic anhydride-alt-acrylic acid] (poly(MA-alt-AA) or MAAA) copolymer was synthesized by free-radical chain polymerization reaction, in 1,4-dioxane in the presence of benzoyl peroxide, 0.1 %, as the radical initiator at 70 °C under a nitrogen atmosphere. The purified copolymer was then modified with an anti-external fungal and anti-cancer active agent, acriflavine (AF). The modification reaction was performed 48 h at 70 °C in dimethylformamide organic media, using triethylamine (TEA or Et₃N) as the catalyst. The modified or conjugated copolymer/drug couple was named as MAAA/AF. Detailed structural characterization of the copolymer (MAAA) and modified product (MAAA/AF) was carried out by Fourier Transform Infrared (FTIR) and Nuclear Magnetic Resonance (¹H-NMR and¹³C-NMR). The obtained FTIR,¹H-NMR, and¹³C-NMR spectra confirmed that AF was successfully bound to the MAAA copolymer backbone by ring-opening reaction. Antimicrobial susceptibility of the MAAA, AF, and MAAA/AF was evaluated by Kirby Bauer Disc Diffusion method on Mueller–Hinton Agar using Enterecoccus faecium, Enterohaemorrhagic Escherichia coli (EHEC), Staphylococcus aureus and Listeria monocytogenes. Results obtained indicated that MAAA/AF had antibacterial activity on EHEC and S. aureus at 50, 40, and 30 µg. A mechanism for MAAA and AF was then also suggested for the conjugation reaction.