The PAMAM dendrimer was synthesized using a multistep approach. To start with, methyl acrylate (2.60 ml, 28.80 mmol) was added dropwise into the methanolic solution (20.00 ml) of propargylamine (0.44 g, 8.00 mmol). The mixture solution was stirred continuously under argon protection for 48 hours, and the temperature was set at 30°C. The solvent was removed via evaporation when the reaction was complete, and the remaining substance was then subjected to silica gel column chromatography using the mixture of petroleum ether and ethyl acetate (EtOAc) (2/1), and the purified product was denoted as precursor 1. The precursor 1 (0.46 mg, 2.02 mmol) was subsequently solubilized in methanol (1.00 ml), into which ethylenediamine (2.50 ml, 37.50 mmol) was added for reaction. The solution was protected using argon atmosphere and stirred for 48 hours at 30°C. The solvent was also evaporated to obtain the solid residue, which were then precipitated three times with the mixture solution of CH3OH and Et2O for further purification. The oil-like product was the amine-terminated precursor 2 (0.55 g, 1.99 mmol), which was further dissolved in methanol (20.00 ml) and mixed with methyl acrylate dropwise (1.25 ml, 13.80 mmol). Argon atmosphere was used to protect the mixture solution, which was stirred at 30°C for 72 hours. The desired product was recovered through evaporation, and impurities were removed using silica gel column chromatography with the mixture solution of EtOAc and CH3OH (10/1), and precursor 3 was thus obtained. Precursor 3 (0.67 g, 1.08 mmol) was subsequently dissolved in methanol (2.50 ml) and again reacted with ethylenediamine (21.40 ml, 21.00 mmol) that was added dropwise under continuous stirring and argon protection. The reaction lasted for 48 hours while the temperature was maintained at 30°C, and the reaction progression was monitored by infrared spectroscopy. When the reaction was finished, the solvent was removed via evaporation, and the residual solids were allowed to precipitate in the mixture of CH3OH and Et2O for three times for complete purification. The PAMAM dendrimers were thus prepared.

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