In the reaction of N³-substituted amidrazones with cis-1,2-cyclohexanedicarboxilic acid anhydride N¹-carbonyl-(2-carboxycyclohexane-1-yl)-N³-substituted-2-picolinamidrazones were obtained.

The chemical structures were confirmed by IR, ¹H NMR, EJ-MS and elemental analysis. Their purity was confirmed by chromatografic methods.

The antiproliferative activity of tested compounds will be assessed in glioma C6 cell culture. Tumor cells will be plated on 96-well microplates (NUNC) at density 0.5 x 10⁴ cells/mL. Next day culture medium will be changed and cells will be exposed to 10 and 100 mM concentrations of tested compounds. Cell proliferation will be assessed after 96 h by means of MTT (Cell proliferation kit I, Roche Diagnostics, Germany) method in which the yellow tetrazolium salt (MTT) is metabolised by viable cells to purple formazan crystals. Microplates will be incubated for 3 h with MMT solution (5 mg/mL). Formazan crystals will be solubilized overnigth in SDS buffer and the product will be quantified spectrophotometrically by measuring absorbance at 570 nm wavelength using E-max Microplate Reader (Molecular Devices Corporation, Menlo Park, CA, USA).

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