Synthesis and activity of NH2- and COOH-terminal elastase recognition sequences on cotton.

The application of peptide recognition sequences of elastase to fibers of wound dressings is a possible route to inhibiting high levels of destructive elastase in the chronic wound. For this reason we have synthesized the elastase recognition sequence Val-Pro-Val on both cotton cellulose, and carboxymethylated cellulose cotton (CMC) and prepared chromatography columns of these to examine elastase retention. The tripeptide was synthesized on cotton-based cellulose fibers both in sequence and as a tripeptide methyl ester. Glycine was employed as a linker of the recognition sequence to the cotton cellulose. Pre-treatment of cotton cellulose with cellulase improved the substitution level of glycine. The peptidocellulose conjugates were employed as a chromatographic stationary phase to assess elastase retention. The sequence Val-Pro-Val-OMe was amino-terminally anchored to carboxymethylated cotton and demonstrated retention of up to 58% of elastase when first applied to the column. Higher repetitive retention was demonstrated subsequently. Cotton gauze similarly modified with Val-Pro-Val-Gly cellulose was compared with untreated gauze for reduction of elastase activity in buffered saline. Solutions of elastase that were treated with Val-Pro-Val-Gly cellulose cotton gauze, demonstrated reduced elastase activity. This study demonstrates the use of elastase recognition sequences as sequestering agents of elastase when attached to cotton fibers and constitutes a model for the design of peptidocellulose analogs in dressing fibers for chronic wounds.