PH_II_P: Pharmaceutical Sciences II. Posters
Zsófia Szász, Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest
Introduction: One of the great possibilities of drug development is the halogenation of molecules, in particular the synthesis of fluorinated derivatives. It is known from the literature that the pharmacokinetic and physicochemical properties of the molecule also change upon fluorination. In addition, fluorinated derivatives are able to target the p53 pathway more effectively than parent molecules.
Aim and methods: The main aim of our present work was to compare the effects of two fluorinated derivatives of TIC-10 (TBP-134, TBP-135) on pancreatic adenocarcinoma (PANC-1) cell line. Tumor selectivity of the drugs was determined on normal human dermal fibroblast (NHDF) cells as a control. The methods used were: (i) measurement of cell viability and cytotoxicity: determination of IC50 values (xCELLigence SP; AlamarBlue assay), measurement of lactate dehydrogenase (LDH) activity (CyQUANT); (ii) monitoring the cell death: determination of Annexin V / 7AAD positive cells (FACSCalibur); (iii) analysis of cell cycle changes (FACSCalibur); (iv) measurement of p53 gene expression (CFX 96 real-time PCR).
Results: The IC50 values of TBP-134 and TBP-135 could be determined after 72 hours, while in the case of TIC-10 treatment only after 96 hours of treatment (IC50: 0.35 and 1.8 μM vs. IC50: 1.8 μM). In our cytotoxicity study, we examined the possible increased LDH activity of the cells, during which the TBP-134 molecule proved to be the most toxic at a concentration of 25 μM. TBP-134 (0.5 μM) significantly increased the number of the early apoptotic cells as early as 24h, whereas TIC10 and TBP-135 were only able to induce apoptosis at 72h. In the cell cycle assay, a G2/M phase block was detectable for all three molecules after 72h. The gene expression of the apoptotic p53 protein was increased 6.8-fold by TBP-134, 6.1-fold by TIC-10, and 3.2-fold by TBP-135 compared to the medium control. As a result of the viability measurement on NHDF cells, it was found that although the molecules reduced the viability of the cells, the IC50 value could not be determined.
Conclusion: Based on our structure-activity studies, the TBP-134 molecule proved to be a more potent derivative on PANC-1 cells than the TBP-135. Furthermore, the position of the introduced fluorine molecule may be an important factor in the antitumor effect.
Funding: EFOP-3.6.3-VEKOP-16-2017-00009; ÚNKP-20-2-SE-16
Semmelweis University, Doctoral School of Pharmaceutical Sciences