Different dextran esters with various degrees of substitution (1, 2 and 3) were synthesized by esterification reaction, with three acid anhydrides: acetic anhydride, propionic anhydride, and butyric anhydride, separately. These modified polysaccharides were characterized by FT-IR, 1H NMR and 13C NMR spectroscopies. Enzymatic degradation of biopolymers by dextranase was also studied. The polymers showing the best degradation profiles were chosen to design blended free films in combination with a polymethacrylate (Eudragit® RS 30D) as a sustained release system for targeting to the colon. These free films were evaluated by permeability of theophylline used as tracer in different in vitro media of the gastro intestinal tract, in presence or in absence of dextranase. From these studies, it was concluded that dextran esters having the lower degree of substitution and constituted of short carbohydrate chains showed the best and significant enzymatic degradation and could be used as a promising carrier for specific colon drug delivery system.Keywords: Colon-Specific Drug Delivery; Polysaccharides; Dextran; Dextranase; Dextran esters; Enzymatic Degradation; Eudragit® RS 30D; Sid-by-side diffusion cel