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The synthesis method of the Cagrilintide comprises the third step of preparing 200mL of cracking reagent, adding the full-protection peptide under an ice bath condition, returning to room temperature after 0.5h, continuing to react for 2h, adding anhydrous ether for precipitation after the reaction is finished, performing centrifugal precipitation for multiple times, adding 300mL of ether each time, and drying to obtain the Cagrilintide product

The best binding was observed for the histidine containing peptide as expected, whereas slightly weaker affinity was observed for amide (Q, N) or amine (K) containing side chain, whereas carboxy (D,E), thioether (M) or aromatic (F,Y,W) functional groups hardly enhanced affinity compared to the single histidine containing peptide (Fig