Membrane proteins are proteins that are embedded in biological membranes and play an important role in various cellular functions. One such protein is the Na+/Ca2+ exchanger, which is an ion transport protein for calcium transport. This exchanger is also found in the inner membrane of the mitochondria (mitochondrial Na+/Ca2+ exchanger). Since intracellular calcium homeostasis is extremely important for cell function, mitochondrial Na+/Ca2+ exchanger inhibitors can cause cell death or apoptosis due to the accumulation of calcium inside the mitochondria. Therefore, the mitochondrial Na+/Ca2+ exchanger is an interesting target for the development of anticancer agents.
The purpose of this Masterʹs thesis was to synthesize and evaluate potential inhibitors of the mitochondrial Na+/Ca2+ exchanger. We used an already known inhibitor CGP37157 to which we attached a triphenylphosphnium cation to achieve a selective targeting of mitochondria. CGP37157 is common to all final compounds, while they differ in the chain n, which was modified by the formation of esters of different lengths, and in the chain m, which was modified by attachment of the corresponding (aminoalkyl)triphenylphosphonium fragment. The progress of the reactions was monitored by thin layer chromatography, the structure of the final compounds was determined by nuclear magnetic resonance and high-resolution mass spectrometry and their purity was checked by high performance liquid chromatography. The final compounds were tested for their anticancer activity on the mouse melanoma cell line B16F10 using the resazurin assay and then compared with the compound UL-EID-2. To verify the selective effect of the final compounds on cancer cells, the assay was also performed on healthy cell lines of mouse myoblasts C2C12 and mouse fibroblasts L929. Compound 9 has the strongest and selective inhibitory activity and it differs from UL-EID-2 by the m chain, which is one C-atom longer in compound 9. Compound 15 has the weakest inhibitory activity and it differs from UL-EID-2 by the length of chain n, which is two C-atoms longer in compound 15. We can conclude that lengthening of the chain m has beneficial effect on the inhibitory activity, while lengthening of the chain n reduces it. These results confirm the inhibitory activity of the synthesized compounds and indicate the meaningfulness of the method for selective delivery to mitochondria and the sense of combining their structural elements.
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