Trypsin‐Catalyzed Kinetically Controlled Synthesis of a Precursor Dipeptide of Thymopentin in Organic Solvents, Using a Free Amino Acid as Nucleophile

Abstract

Trypsin‐catalyzed, kinetically controlled synthesis of a precursor, dipeptide of thymopentin (TP‐5), Bz–Arg‐Lys–OH (or–OEt) in organic solvents was studied. Bz–Arg–OEt was used as the acyl donor and Lys–OH and Lys–OEt were used as the nucleophiles. Ethanol was selected as the organic solvent from ethanol, methanol, acetonitrile, and ethyl acetate tested under the experimental conditions. As expected, Lys–OEt is not a suitable nucleophile in trypsin‐catalyzed reaction, due to its competition with the protective Arg–OEt as acyl donor for the active site of trypsin, while Lys–OH does not have this problem. The optimal reaction condition for the synthesis of Bz–Arg‐Lys–OH was set up as 20% Tris‐HCl buffer, pH 8.0, 35°C for 6 h with the yield of 52.5%, or for 18–24 h with the yield of about 60%.