Abstract:  Aspartimide formation is one of the most common secondary reactions on solid phase peptide synthesis. In the present work, we describe the optimization of the synthesis of two thrombospondin fragments containing an Asp‐Gly sequence that show a strong tendency to form cyclic aspartimide derivatives in an unusual high percentage. Several different strategies were applied changing type of resin, Fmoc‐deprotection reagents, coupling additives, resin cleavage cocktails and the use of Hmb‐Gly derivative to minimize the extension of this byproduct. Best results were obtained with cross‐linked ethoxylate acrylate (CLEAR®‐cross‐linked ethoxylate Acrylate Resin)‐type resin and pip/dimethylformamide deprotection. Besides, as in biological assays the aspartimide containing sequence resulted to be more active than the linear one, the optimization of its synthesis was also carried out.