Antiproliferative Effect of Novel Aminoacridine-based Compounds

Abstract

We tested the antiproliferative activity and mechanism of the action of several novel aminoacridine derivatives. Six different cancer cell lines were used to evaluate the potential cytotoxic effect of eleven aminoacridine-based molecules. A standard MTT assay was used for cell bioavailability analysis. Additionally, the potential cytotoxic effect of the tested compounds on non-cancer cells was investigated in rat skeletal muscle myotubes (L6) and in bovine aortic smooth muscle cells. In order to investigate whether the DNA binding activity of tested compounds correlated with their cytotoxic effect, circular dichroism (CD) measurement and DNA T4 ligase assay were performed. Finally, the potential mutagenic activity of the lead compound 5 was investigated. The cytotoxic effect of compound 5 in cancer cells was obtained in lower concentrations than the well-known: 9- aminoacridine based drug, amsacrine. The lead compound binds to DNA, but in a different mode than the parent molecules. Additionally, compound 5 was not cytotoxic in the effective range of concentrations in non-cancer cells. In identical concentrations, the parent compound (9-aminoacridine) and amsacrine were extremely toxic for both types of these normal cells. Finally, based on CD measurement and T4 ligase assay, it was confirmed that 5 binds to DNA but in different from the parent compounds manner. Important to mention, that compound 5 might have increased mutagenic activity which must be verified in vivo. Based on these in vitro results, we conclude that 5 is a more potent and more selective antiprolifirative compound than amsacrine. Compound 5 was also more effective in HepG2 and P-12 cells. Thus, 5 is suitable for future in vivo biological evaluation and its structure might be used as a basis for developing novel anticancer drugs.

Mimicking Neuroligin-2 Functions in β-Cells by Functionalized Nanoparticles as a Novel Approach for Antidiabetic Therapy

“…We described this test in our previous publication . Briefly, cells were incubated with MT reagent (2 mg/mL) in growth medium for 30 min at 37 °C.…”

“…INS-1E cells were grown in DMEM (22.5 mM glucose) supplemented with 10% fetal calf serum (FCS), 1 mM glutamine, and antibiotics (100 μg/mL penicillin, 100 μg/mL streptomycin) at 37 °C in a 5% CO 2 humidified atmosphere, as described by Pasternak et al PC-3 and PC-12 were grown and maintained as we previously described …”

“…We described this test in our previous publication. 64 Briefly, cells were incubated with MT reagent (2 mg/ mL) in growth medium for 30 min at 37 °C. The medium was then aspirated, and DMSO was added to solubilize the cells and colored crystals.…”

“…INS-1E cells were grown in DMEM (22.5 mM glucose) supplemented with 10% fetal calf serum (FCS), 1 mM glutamine, and antibiotics (100 μg/mL penicillin, 100 μg/mL streptomycin) at 37 °C in a 5% CO 2 humidified atmosphere, as described by Pasternak et al 66 PC-3 and PC-12 were grown and maintained as we previously described. 64 Cell Lysate Preparation. Whole cell lysates were prepared as previously described 67 with some minor modifications: the lysis buffer contained 50 mmol/L Tris-HCl, pH 7.5, 1 mmol/L EDTA, 1 mmol/L EGTA, 1 mmol/L Na 3 VO 4 , 150 mmol/L NaCl, 50 mmol/L NaF, 10 mmol/L sodium-glycerophosphate, 5 mmol/L sodium pyrophosphate, and 1 mmol/L PMSF, supplemented with 0.1% (v/v) IGEPAL, 0.1% (v/v) 2-β-mercaptoethanol, and protease inhibitor cocktail (1:100 dilution).…”

Structural patterns enhancing the antibacterial activity of metallacarborane-based antibiotics

“…showed surprisingly low cytotoxicity, despite being derived from fairly toxic 9-aminoarcridine 38 . Thus, arene-based organic substituents resulted in derivatives with no antibacterial activity but different toxicities.…”