An eco-friendly methodology for preparing Fmoc-, Z-, and Boc-Nprotected dipeptides and tripeptides is described, from the corresponding Nprotected-α-aminoacyl benzotriazoles and α-amino acid derivatives, with different Cterminal functionalities such as esters or amides, using vibrational ball-mill (VBM). The reactivity of a β-amino ester was also investigated. In some cases, the coupling was achieved by liquid-assisted grinding (LAG). α,αand one α,β-dipeptide were obtained in good to excellent yields mainly by precipitation in water, resulting in an improved environmental impact compared to classical peptide synthesis in solution, as shown by green metric calculations. The method was extended to the biotinylation, via an aminohexanoyl spacer, of the pentapeptide RGDfV, which contains the well-known integrins recognition site arginine−glycine−aspartic acid (RGD) motif.