Notably, the macrocyclic content in the aged mixtures is significantly affected by milling duration, when monomer shuffling occurs. Thus, aging of the short-milled (5 minutes) reaction mixture, which contained mainly homomeric CU oligomers, resulted in the ripening of cycHC [8] as the dominant product (55% yield), along with a minor quantity of mixHC [8] (16% yield). However, fast reversible C-N bond formation and cleavage during milling caused rapid dynamic changes in the oligomeric profile with a random distribution of the biotin units. Upon prolonged milling (60 min, see Tables ), the yield of mixHC [8] notably increased from 16% to 37%, with a concurrent decrease in the yield of cycHC [8] to 38%, resulting in a 1:1 product ratio. Finally, close examination of the aging duration (Table ) revealed that the maximum content of mixHC [8] was achieved in 3 hours, when the self-assembly process was essentially complete.