A series of polyurethane elastomers (PUEs) derived from three thiacalix[4]arene derivatives (TC4As), namely p-tert-butylthiacalix[4]arene, tetrasodium thiacalix[4]arenetetrasulfonate and thiacalix[4]arenetetrasulfonic acid, as a portion of chain extender in a mixture with glycerol were synthesized. The effects of the chemical structure of TC4As used as chain extenders on the various properties of the prepared PUEs were investigated and compared with PUE extended with only glycerol as chain extender using Fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray diffraction (XRD), scanning electron microscopy and a universal tensile tester. Moreover, the effect of the introduction of TC4As as a portion of chain extender on the hydrophobicity of the PUEs was also evaluated. DSC, FTIR spectroscopy and XRD revealed that the degree of phase separation and crystallinity in TC4A-based PUEs was much higher than that of the glycerol-based ones. Thus, it was concluded that the presence of TC4As in TC4A-based PUEs seems to favour the formation of a more ordered structure due to an increase in the degree of phase separation. The TGA results also showed that, with incorporation of TC4As into the polyurethane backbone, the thermal stability of PUEs was improved.