Fast ill Situ forming supramolecular hydrogels consisted of the tyramine-conjugated supramolecular structures and chitosan derivative were prepared via an enzymatic reaction with horseradish peroxidase (HRP) and hydrogen peroxide (H2O2). The gel formation was varied within a time period of 5 s to 10 min by controlling the concentrations of HRP, H2O2, and polymers. Tyramine conjugation at different sites of the supramolecular structure resulted in significant changes in physical properties and the degradation time of the hydrogels that were confirmed by water uptake, compressive strength and degradation tests. In addition, the hydrogels showed it good cytocompatibility in vitro, These hydrogels could be promising injectable biomaterials with adjustable degradation times to control both the cellular behaviors as a regenerative cell matrix and the drug release behavior as a drug delivery vehicle.