A novel efficient synthetic method for regioselective synthesis of optically active 2-mercaptoimidazoles with three points of structural diversity was investigated on a polyethylene glycol (PEG) support. The key synthetic steps involve (i) synthesis of thiourea derivatives of polymer-supported amino acids with isothiocyanates and (ii) one-pot regioselective condensation of PEG-linked thiourea with α-bromo ketones to furnish the 2-mercaptoimidazole skeleton under microwave conditions. An excellent regioselectivity was observed during this one-pot condensation reaction which was further supported by NOE studies. In addition to three sets of structural diversity, supplementary chirality at the α-position of the 2-mercaptoimidazole skeleton is the key feature of this synthesis. A representative set of 2-mercaptoimidazoles was efficiently assembled on soluble support utilizing various L-amino acids, isothiocyanates and α-bromoaryl ketones with good yields.