Solvent‐Free Mechanosynthesis of Oligopeptides by Coupling Peptide Segments of Different Lengths – Elucidating the Role of Cesium Carbonate in Ball Mill Processes

We report an idea for the synthesis of oligopeptides using a solvent‐free ball milling approach. Our concept is inspired by block play, in which it is possible to construct different objects using segments (blocks) of different sizes and lengths. We prove that by having a library of short peptides and employing the ball mill mechanosynthesis (BMMS) method, peptides can be easily coupled to form different oligopeptides with the desired functional and biological properties. Optimizing the BMMS process we found that the best yields we obtained when TBTU and cesium carbonate were used as reagents. The role of Cs2CO3 in the coupling mechanism was followed on each stage of synthesis by 1H, 13C and 133Cs NMR employing Magic Angle Spinning (MAS) techniques. It was found that cesium carbonate acts not only as a base but is also responsible for the activation of substrates and intermediates. The unique information about the BMMS mechanism is based on the analysis of 2D NMR data. The power of BMMS is proved by the example of different peptide combinations, 2+2, 3+2, 4+2, 5+2 and 4+4. The tetra‐, penta‐, hexa‐, hepta‐ and octapeptides obtained under this project were fully characterized by MS and NMR techniques. We present a concept inspired by playing with blocks, in which it is possible to construct various objects from segments (blocks) of different sizes and lengths. An attempt is made to explain the mechanism of coupling reaction in a ball mill in the presence of cesium carbonate. The usefulness of the method in the synthesis of oligopeptides has been proven.