A series of 2,6-diarylpiperidin-4-ylidene thiazolidin-4-one derivatives (15–21) were efficiently synthesized using a green and recyclable catalyst Amberlit IR-120H resin under microwave irradiation. All the compounds were characterized by elemental analysis, FT-IR, 1H, 13C NMR spectra. The conformation of the compound was unambiguously assigned by 2D NMR spectral analysis. Single crystal X-ray diffraction recorded for compound 21 and the data indicate that the compound crystallized in triclinic system with space group Pī. The observed chemical shifts and coupling constants suggested that the piperidone ring (15–21) adopts in chair conformation with equatorial orientations of the aryl groups. In solid state the configuration about C[dbnd]N double bond is syn to C-5 carbon. All the compounds were screened for their in vitro antibacterial and antimycobacterial activities. Among the tested, compounds 17 and 18, were found to be potent inhibition with MIC values in the range of 0.2–0.8 μg/mL. The interactions of the compounds with calf thymus DNA were investigated by spectroscopic method and the results indicated that compounds bound to Deoxyribo Nucleic Acid (DNA) by groove binding mode. © 2019 Elsevier B.V.