New active poly(ethylene glycol) derivative for amino coupling

Abstract The synthesis and characterization of an active poly(ethylene glycol) (PEG) derivative with new properties has been afforded starting from a side reaction of N -hydroxysuccinimide (NHS) in presence of N,N -dicyclohexylcarbodiimide (DCC). In particular, a ring opening of NHS in presence of DCC forms a β-alanine active derivative, through Lossen rearrangement, which then reacts with PEG–NH 2 yielding the active PEG derivative: PEG–NH–CO–βAla–NH–CO–NHS. The active group R–NH–CO–NHS showed lower reactivity towards amines and a higher stability in alkaline solution. This can be especially useful in polymer coupling to proteins, because it may lead to a better selectivity among all the amino groups present in a protein, thus yielding less heterogeneous PEG-protein conjugates mixture because only the most nucleophilic and solvent exposed amines can react. A comparison with one of the most used PEGylating agent, the PEG-succinimidyl carboxymethylate (PEG–O–CH 2 –CO–NHS), displayed the usefulness of this new PEG derivative for obtaining PEG-protein conjugates. Furthermore, it is of interest that the obtained protein-conjugates are slowly hydrolyzed in water releasing the free protein, therefore they can be considered protein prodrugs.