Anisole and 1,3-dimethoybenzene are excellent alternatives to DCM for cleaving protected peptides.

Solid-phase peptide synthesis (SPPS) is the method of choice for the preparation of peptides in both laboratory scale and large production. Although the methodology has been improved during the last decades allowing the achievement of long peptides and challenging sequences in good yields and purities, the process was not revised from an environmental point of view. One of the main problems in this regard is the large amount of solvents used, and therefore the tons of generated waste. Moreover, the solvent of choice for the SPPS is N,N-dimethylformamide (DMF), which is considered as reprotoxic; thus, there is an urgent necessity to replace it with safer solvents. The DMF substitution by a green solvent is not a trivial task, because it should solubilize all the reagents and byproducts involved in the process, and, in addition to facilitating the coupling of the different amino acids, it should not favor the formation of side-reactions compared with DMF. Herein, it was demonstrated that the use of the green solvent N-butylpyrrolidinone (NBP) as a replacement of DMF was beneficial in two well-documented side reactions in peptide synthesis, racemization and aspartimide formation. The use of NBP rendered a lower or equal level of racemization in the amino acids more prone to this side reaction than DMF, whilst the aspartimide formation was clearly lower when NBP was used as solvent. Our findings demonstrate that the use of a green solvent does not hamper the synthetic process and could even improve it, making it environmentally friendlier and synthetically better.

NBP has proved an excellent alternative solvent to the hazardous DMF for SPPS.