ILKA molecules are nationally developed cannabinoid and bixin containing derivatives with expected antioxidant effects.

The aim of our study was to investigate whether pretreatment with ILKA 675 and ILKA 676 molecules has a protective effect against hypoxia-reoxygenation (H/R) induced acute injury on cardiac myocytes.

In our series of experiments the H9c2 rat cardiomyoblast cells were treated for 24 hours and the IC50 values of the molecules were determined. After that, we investigated the cell viability using MTT assay. The concentration of 30 µM proved to be the most effective. Subsequently, this concentration was used to pretreat cardiomyocytes followed by four/three hours of H/R. The LDH assay was carried out to give information regarding the cytotoxicity of the molecules. Furthermore, antioxidant assays such as TEAC (Trolox Equivalent Antioxidant Capacity), FRAP (Ferric Reducing Antioxidant Power) and ORAC (Oxygen Radical Absorbance Capacity) were carried out to evaluate the antioxidant properties of the molecules.

Our findings demonstrate that the pretreatment with the newly synthesised molecules significantly improved the cardiomyocytes viability after H/R. ILKA 675 was the least cytotoxic under hypoxic conditions. Furthermore, ILKA 675 showed the highest antioxidant activity according to the results obtained from the antioxidant assays.

Overall, ILKA 675 and ILKA 676 may have a beneficial effect in the prevention of H/R induced cytotoxicity through the reduction of oxidative stress.