In this study, we report the synthesis of glyoxylamide peptide-mimics as self-assembled gels with well-defined molecular structures for topical delivery of ciprofloxacin (CIP). The glyoxylamide peptide mimics successfully formed hydrogels with critical gel concentrations of 0.02-0.08% (w/v). The mechanical strength, secondary structure, and fiber morphology of these hydrogels can be modulated by varying the N-substituent of the ring-opened isatins. The synthesised hydrogel exhibited a high loading capacity of CIP (40% (w/w)) and a sustained release profile. The CIP-loaded hydrogels were able to release CIP for more than 15 days and the released solution was shown to retain activity against Gram-positive and Gram-negative bacteria. In addition, the hydrogels formed showed low toxicity against Cos7 cells.