Reactive Oxygen Species Production Is Responsible for Antineoplastic Activity of Osmium, Ruthenium, Iridium and Rhodium Half-Sandwich Type Complexes with Bidentate Glycosyl Heterocyclic Ligands in Various Cancer Cell Models

Platinum complexes are used in chemotherapy, primarily as antineoplastic agents. In this study, we assessed the cytotoxic and cytostatic properties of a set of osmium(II), ruthenium(II), iridium(III) and rhodium(III) half-sandwich-type complexes with bidentate monosaccharide ligands. We identified 5 compounds with moderate to negligible acute cytotoxicity but with potent long-term cytostatic activity. These structure-activity relationship studies revealed that: 1) osmium(II) p-cymene complexes were active in all models, while rhodium(III) and iridium(III) Cp* complexes proved largely inactive; 2) the biological effect was influenced by the nature of the central azole ring of the ligands—1,2,3-triazole was the most effective, followed by 1,3,4-oxadiazole, while the isomeric 1,2,4-oxadiazole abolished the cytostatic activity; 3) we found a correlation between the hydrophobic character of the complexes and their cytostatic activity: compounds with O-benzoyl protective groups on the carbohydrate moiety were active, compared to O-deprotected ones. The best compound, an osmium(II) complex, had an IC50 value of 0.70 µM. Furthermore, the steepness of the inhibitory curve of the active complexes suggested cooperative binding; cooperative molecules were better inhibitors than non-cooperative ones. The cytostatic activity of the active complexes was abolished by a lipid-soluble antioxidant, vitamin E, suggesting that oxidative stress plays a major role in the biological activity of the complexes. The complexes were active on ovarian cancer, pancreatic adenocarcinoma, osteosarcoma and Hodgkin’s lymphoma cells, but were inactive on primary, non-transformed human fibroblasts, indicating their applicability as potential anticancer agents.

Allelopathic potential and cytostatic activity of Antarctic moss Sanionia georgicouncinata (Amblystegiaceae)

From the first half of the XX century, the maritime Antarctic was a region with high regional warming rates. At the beginning of the XXI century, regional warming strongly slowed down, but during the next twenty years the near-surface temperatures and mean annual precipitation at the Antarctic Peninsula will most probably increase. Higher temperatures and precipitation make the conditions in the maritime Antarctic more favourable for plants, including invasive species. Moreover, the increasing number of Antarctic tourists unintentionally transport thousands of seeds to the Antarctic on their clothes and equipment. In such circumstances, it is important to evaluate possible defensive mechanisms of native plant species, including their allelopathic potential. The objective of this study was to evaluate the allelopathic potential and cytostatic activity of Sanionia georgicouncinata. This species is widespread in the maritime Antarctic and is one of the most abundant moss species on the Argentine Islands (Wilhelm Archipelago). Seeds of cucumber Cucumis sativus, radish Raphanus raphanistrum subsp. sativus, wheat Triticum vulgare, and amaranth Amaranthus paniculatus were used as bioassay species. Fresh green biomass of S. georgicouncinata was used to prepare aqueous extracts with 1 : 10, 1 : 50, and 1 : 100 plant to water ratios. The average root length of wheat sprouts and cucumber sprouts in distilled water did not differ significantly from those in plant extracts. The root length of radish sprouts in 1 : 50 plant extract was significantly shorter than in distilled water. Meanwhile, in 1 : 50 and 1 : 100 water extracts, roots of amaranth plants were significantly longer than in water. The number of lateral roots of cucumber sprouts in water and studied aqueous extracts varied from 1 to 6 with medians from 3 to 4. The medians of control (distilled water) and sample groups (aqueous extracts) did not differ significantly, but only 38.1 ± 6.1% of cucumber sprouts developed lateral roots in the 1 : 10 aqueous extract of S. georgicouncinata, and even less – 28.3 ± 7.5% in the 1 : 50 aqueous extract. The results of this study show that the allelopathic potential of S. georgicouncinata is low. However, a significant cytostatic activity of the studied extracts shows that this species may still affect other plants and animals. Further research is needed to evaluate the impact of the studied extracts on root growth and germination of the invasive plants, particularly Poa annua.

Оценка эффективности противоопухолевого воздействия и индукции апоптоза в клетках карциномы почки человека биологически активными добавками, содержащими ресвератрол, индол-3-карбинол и кордицепин, флуоресцентными методами визуализации

We compared the effectiveness of antitumor effects and apoptosis induction on human renal cell carcinoma A498 extracts of cruciferous plant material (indole-3-carbinol), Chinese mushroom (cordycepin), French red wine (resveratrol) at low concentrations after 24 and 48 h using fluorescent methods of imaging apoptosis and necrosis in human tumor cells in vitro . Propidium iodide and acridine orange were used as dyes in the "alive and dead" test, which allowed us to detect the number of dead cells and cells in which apoptosis had started. It was found that indole-3-carbinol at low concentrations (0.0288 and 0.1152 mg/ml) has a pronounced cytotoxic and cytostatic activity against human kidney cancer cells, significantly exceeding the action of resveratrol at the same concentrations. At the same time, cordycepin has no cytotoxic and cytostatic activity at these concentrations. Indole-3-carbinol also has the most pronounced apoptotic activity at concentrations of 0.0144-0.1152 mg/ml, after 48 h the number of kidney cancer cells in apoptosis increases by 6.8-10 times compared to control. It is concluded that indole-3-carbinol is a promising antitumor agent for use in the complex therapy of patients with kidney cancer.

Magnetically sensitive nanocomposites for targeted antitumor therapy with application of gemcitabine

The aim of the work is synthesis and study on the properties of polyfunctional magnetosensitive nanocomposites (NC) and target-directed magnetic fluids (MF) based on physiological solution (PS), magnetite, gemcitabine (GEM) and HER2 antibodies (AB), promising for use in targeted antitumor therapy against MDA-MB-231 aggressive tumor cells of triple-negative human breast cancer (BC) with high proliferative and metastatic activity. The specific surface area (Ssp) of samples was determined by the method of thermal desorption of nitrogen using a device KELVIN 1042 of “COSTECH Instruments”. The size of nanoparticles (NP) has been estimated by the formula DBET = 6/(ρSBET), where ρ is the density of NC particle, SBET is the value of the specific surface area calculated by the polymolecular adsorption theory of Brunauer, Emmett and Teller (BET). The surface condition of nanodispersed samples was studied by IR spectroscopy (“Perkin Elmer” Fourier spectrometer, a model 1720X). To calculate the concentration of hydroxyl groups on the surface of nanostructures, the method of differential thermal analysis was used in combination with differential thermogravimetric analysis. The thermograms were recorded using a derivatograph Q-1500D of MOM firm (Hungary) in the temperature range of 20–1000 °C at a heating rate of 10 deg/min. X-ray phase analysis of nanostructures was performed using a diffractometer DRON-4-07 (CuKα radiation with a nickel filter in a reflected beam, the Bragg-Brentano focusing). The size and shape of NP were determined by electron microscopy (a transmission electron microscope (TEM) JEM-2100F (Japan)). The hysteresis loops of the magnetic moment of the samples were measured using a laboratory vibration magnetometer of Foner type at room temperature. Measurement of optical density, absorption spectra and GEM concentration in solutions was performed by spectrophotometric analysis (Spectrometer Lambda 35 UV/Vis Perkin Elmer Instruments). The amount of adsorbed substance on the surface of magnetite was determined using a spectrophotometer at λ = 268 nm from a calibration graph. The thickness of the adsorbed layer of GEM in the composition of Fe3O4@GEM NC was determined by magnetic granulometry. To study the direct cytotoxic/cytostatic effect of a series of experimental samples of MF based on PS, Fe3O4 NP, GEM, HER2 AB, as well as MF components in mono- or complex use, onto MDA-MB-231 cells in vitro, IC50 index was determined. MF were synthesized on the basis of single-domain Fe3O4 and PS, stabilized with sodium oleate (Ol.Na) and polyethylene glycol (PEG), containing GEM and HER2 (Fe3O4@GEM/Ol.Na/PEG/HER2+PS). The cytotoxic/cytostatic activity of MF against MDA-MB-231 cells was studied. It was found that as a result of application of synthesized MF composed of Fe3O4@GEM/Ol.Na/PEG/HER2+PS at the concentration of magnetite of 0.05 mg/mL, GEM - 0.004 mg/mL and HER2 AB - 0.013 μg/mL, a synergistic effect arose, with reduction of the amount of viable BC cells to 51 %. It has been proved that when using MF based on targeted Fe3O4/GEM/HER2 complex, the increased antitumor efficacy is observed compared to traditional use of the drug GEM, with a significant reduction (by four times) of its dose. The high cytotoxic/cytostatic activity of Fe3O4/GEM/HER2 complexes is explained by the fact that endogenous iron metabolism disorders play a significant role in the mechanisms of realization of the apoptotic program under the influence of nanocomposite. Thus, when the nanocomposite system contains Fe3O4/GEM/HER2 complexes in MDA-MB-231 cells, a significant increase is observed in the level of “free iron”, which favours formation of reactive oxygen species and causes oxidative stress (Fenton reaction). The consequences of oxidative stress are induction of apoptosis, enhancement of lipid peroxidation processes, as well as structural and functional rearrangement of biological membranes. The prospects have been shown of further studies of Fe3O4@GEM/Ol.Na/PEG/HER2+PS MF in order to create on their basis a magnetically carried remedy for use in targeted antitumor therapy.