Effects of Novel Chalcone Derivatives upon H9c2 and MDCK Cell Viability

Introduction: The characteristic 3’4’5’-trimethoxy aromatic ring from Combretastatin A-4 has been utilised in many chalcone (1,3-diphenyl-2-propen-1-one) derivatives to produce promising anticancer activity (1). Despite their observed efficacy (2), many chalcone-based drug discovery studies have not addressed cardiovascular toxicity. However, this is a significant adverse effect associated with many anticancer drugs, hence, we looked at the effects of novel chalcone derivatives upon 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) reduction using H9c2 and MDCK cell lines as models of cardiac and renal cells, respectively.

Synthesis: Potassium Hydroxide 3 ml (20% w/v solution) was added to a mixture of 3’4’5’-trimethoxyacetophenone (5 mmol) and aryl aldehyde (5 mmol) dissolved in 9 ml of ethanol. The mixture was stirred at room temperature for 1 hour monitored by thin layer chromatography. The precipitate formed was filtered off and washed with 2x 30ml water and 2x 30ml cold ethanol. Purification via ethanol recrystallization was used to produce pure chalcone which was verified by NMR, IR and MS (3).

MTT assay: H9c2 and MDCK cells were cultured in complete Dulbecco’s Modified Eagle’s Medium (DMEM) and maintained in an incubator (37°C, 5 % CO2). For experiments, cells were seeded into 96-well plates (5000 cells/well) and, 24 hours later, treated with novel compounds (2 hours incubation at 37°C, 3-1000 μM in serum-free DMEM). Cell viability was assessed by the colorimetric conversion of (MTT) to purple formazan product (measured at 560 nm). Analysis was performed by one-way ANOVA with Dunnet’s multiple comparisons test against control (Serum-free DMEM only). Data are expressed as mean % of maximum inhibition ± standard error of the Mean, P<0.05 vs. control.

Results: All four novel chalcone compounds produced concentration-dependent inhibition of cell viability, as measured by MTT reduction, in both H9c2 cells (n=4) and MDCK cells (n=5).

Conclusions: These preliminary data suggest that MK33 shows most toxicity in both cell types; however, inhibition reached significance at 100μM in H9c2 cells and 30μM in MDCK cells. Further studies will be completed to assess the effects of these compounds upon cancer and endothelial cell viability leading to structure activity relationship analysis.

References:

1. Ducki S et al. (2009). Biorganic and medicinal chemistry 17: 7698-7710.

2. Mahapatra DK et al. (2015). European Journal of Medicinal Chemistry 98: 69-114.

3. Detsi A et al. (2009). Biorganic and medicinal chemistry 17: 8073-8085.