A series of controllable amphiphilic block copolymers composed of poly(ethylene oxide) (PEO) as the hydrophilic block and poly(ɛ-caprolactone) (PCL) as the hydrophobic block with the amino terminal group at the end of the PEO chain (PCL-b-PEO-NH2) were synthesized. Based on the further reaction of reactive amino groups, diblock copolymers with functional carboxyl groups (PCL-b-PEO-COOH) and functional compounds RGD (PCL-b-PEO-RGD) as well as the triblock copolymers with thermosensitive PNIPAAm blocks (PCL-b-PEO-b-PNIPAAM) were synthesized. The well-controlled structures of these copolymers with functional groups and blocks were characterized by gel permeation chromatography (GPC) and 1H NMR spectroscopy. These copolymers with functionalized hydrophilic blocks were fabricated into microspheres for the examination of biofunctions via cell culture experiments and in vitro drug release. The results indicated the significance of introducing functional groups (e.g., NH2, COOH and RGD) into the end of the hydrophilic block of amphiphilic block copolymers for biomedical potentials in tissue engineering and controlled drug release.