Final Report Summary - RELOADPEPSYN (A Reloadable Molecular Peptide Synthesizer)
After series of optimization reactions, target machine was synthesized and characterized using modified macrocycle (M) and stopper moieties T1 and T2 as planned in section B4-part 1.2 Scheme 3 of the proposal. Pyridiyl moiety on T1 was replaced with hydrazide in order to be able to load the amino acid via another hydrazone linkage. This optimized design is preferred due to unstable nature of original metal complex in the presence of thiolate. Rotaxane formation using copper-catalysed Huisgen cycloaddition, deprotection of the machine gave decent yields and characterised by various 2D NMR spectra and High Resolution Mass Spectrometry. The machine was loaded with two amino acids under acidic conditions and operated in the presence of base (triethylamine) and reducing agent, Tris(2-carboxyethyl)phosphine hydrochloride. Analysis of the crude operation product by High Resolution Mass Spectrometry indicates that two amino acids are successfully incorporated to the growing chain and Tandem Mass Spectroscopy analysis proves that these amino acids are in correct order. These findings suggest that information on the track is successfully used to load each amino acid to specifıc region of the machine.