Several novel lactones containing either a pyridin-3-yl or a pyridin-2-yl ring in their structure were synthesized: trans δ halolactones 6–8a,b (chloro‑, bromo‑ and iodo-), γ iodolactone 11b and isomeric γ hydroxylactones 9–10a,b. Their structures were confirmed by NMR, IR and HR-MS and additionally by single crystal X-ray diffraction for trans δ bromolactone 6a, trans δ chlorolactone 7b and γ hydroxylactone 10b. Spectral analyses were further supported by DFT calculations. In testing of the compounds for antibacterial activity, none of the lactones exhibited activity against several bacteria strains. However, all lactones exhibited significant cytotoxicity against normal eukaryotic murine fibroblast L929 and human gastric adenocarcinoma AGS cell lines with lactones possessing a pyridine-2-yl ring being more cytotoxic than their pyridine-3-yl counterparts.