Antitumor effects of new derivatives of N-9 sulfonylpurine

Uvod: Tumorske bolesti, kao vodeći uzrok smrtnosti u svijetu, nameću potrebu za pronalaskom učinkovite tumorske terapije. Purinski nukleozidni analozi (PNA) prema dosadašnjem pokazanom farmakološkom djelovanju zaslužuju pažnju i daljnje istraživanje. Cilj istraživanja: Odrediti citotoksični učinak N-9 sulfonilpurina na tumorske i normalne stanične linije i utvrditi mogući način stanične smrti. Materijali i metode: Citotoksični učinak 14 novosintetiziranih N-9 sulfonilpurina utvrđen je na panelu humanih tumorskih stanica (HeLa, CaCo-2, NCI-H358, K562, Raji, HUT-78, CCRF-CEM, MOLT-4), normalnim humanim fibroblastima (BJ) i normalnim epitelnim stanicama psećeg bubrega (MDCK). Vijabilnost stanica nakon tretmana s navedenim spojevima utvrđena je MTT testom, a citotoksični učinak potvrđen je mjerenjem LDH enzima u staničnom mediju. Način stanične smrti istražen je mjerenjem aktivnosti enzima kaspaze 3 u staničnom lizatu, a apoptotski put utvrđen je mjerenjem mitohondrijskog membranskog potencijala protočnom citofluorimetrijom. Rezultati: Testirani N-9 sulfonilpurini pokazuju citotoksičan učinak u ovisnosti o primjenjenoj koncentraciji i staničnoj liniji. Na temelju rezultata izražene citotoksičnosti na humanim staničnim linijama leukemija i limfoma, izdvojeni su spojevi R-191 i D7-23. Navedeni spojevi ne pokazuju aktivaciju kaspaze 3, ali je narušen mitohondrijski membranski potencijal. Zaključak: Iz dobivenih rezultata možemo zaključiti da određeni novosintetizirani N-9 sulfonil purini pokazuju izrazito antiproliferativno i citotoksično djelovanje, osobito na staničnim linijama humanih leukemija i limfoma. Izostanak aktivacije enzima kaspaze 3 nametnuo je smjernicu da se detaljno istraži i potvrdi kaspaza-neovisan put apoptoze.Introduction: Tumor diseases are a leading cause of death worldwide and there is a need to discover an efficient form of tumor treatment. According to recently shown pharmacological activity, purine nucleoside analogs (PNA) deserve attention and further research. Study aim: To determine the cytotoxic effect of N-9 sulfonyl purine on tumor and normal cell lines and to determine a possible mechanism behind cell deaths. Materials and methods: The cytotoxic effect of 14 newly synthesized N-9 sufonyl purines were identified on a panel of human tumor cells (HeLa, CaCo-2, NCI-H358, K562, Raji, HUT-78, CCRF-CEM, MOLT-4), normal human fibroblasts (BJ) and normal epithelial cells of a dog's kidney (MDCK). After the treatment, which involved the previously mentioned compounds, cell viability was determined by a MTT test and the cytotoxic effect was confirmed by measuring the LDH enzyme in the cell culture medium. The mechanism behind cell death was examined by measuring the caspase-3 enzymatic activity in the cell lysate, while the apoptotic pathway was determined by measuring the mitochondrial membrane potential by means of flow cytofluorometry. Results: The examined N-P sulfonyl purine shows the cytotoxic effect depending on the applied concentration and cell line. Based on the results, cytotoxicity was expressed on human leukemia and lymphoma cell lines and compounds R-191 and D7-23 were isolated. The previously mentioned compounds did not show caspase-3 activity, but the mitochondrial membrane potential was distorted. Conclusion: Based on the obtained results we can conclude that certain newly synthesized N-9 sulfonyl purines show high anti-proliferative and cytotoxic activity, especially on the human leukemia and lymphoma cell lines. The absence of caspase-3 activity demands further detailed research and confirmation of a caspase-independent apoptotic pathway