In vitro investigation of antitumor activities of extracts of endemic plant species Helichrysum zivojini Černjavski et Soška U

Bioaktivni sastojci biljnih vrsta nalaze se u centru pažnje istraživanja u savremenoj onkologiji zbog svoje moguće uloge u hemioprevenciji, odnosno inhibiciji različitih koraka procesa maligne transformacije. Antikancerski potencijal biljnih jedinjenja zasniva se na mogućnosti redukcije slobodnih radikala, regulacije karcinogen-aktivirajućih i karcinogen-detoksifikujućih enzima, kao i mogućnosti inhibicije inflamatornih citokina, zatim na mogućnosti da dovedu do promena u regulaciji faktora rasta i ciljnih molekula signalnih puteva koji kontrolišu ćelijski rast, proliferaciju i apoptozu, kao i angiogenezu, invaziju i metastazu malignih ćelija. Osnovni cilj istraživanja je bio da se ispita citotoksičnost, odnosno odrede intenzitet i mehanizmi citotoksičnog dejstva pet ekstrakata izolovanih u vidu frakcija iz endemične biljne vrste Helichrysum zivojinii Černjavski et Soška prema specifičnom malignom ćelijskom tipu, kao i da se odredi selektivnost u antitumorskom dejstvu prema nizu humanih malignih ćelija, kako u odnosu na poreklo - tip tumora, tako i u odnosu na zdrave mononuklearne ćelije periferne krvi (PBMC). Kako bi se doprinelo boljoj proceni antitumorskog potencijala ekstrakata, ispitan je i uticaj ekstrakata na invazivnost humanih metastatskih malignih ćelija i na angiogenezu endotelijalnih ćelija. Hemijska karakterizacija biljnih ekstrakata je imala za cilj da omogući razumevanje veze između kvalitativnog sastava i intenziteta citotoksične aktivnosti ekstrakata. Citotoksična aktivnost pet ekstrakata endemične biljne vrste Helichrysum zivojinii Černjavski et Soška je ispitana na sledećim humanim malignim ćelijskim linijama: HeLa (adenokarcinom cerviksa), Fem-x (melanom), K562 (mijeloidna leukemija), MDA-MB-361 (adenokarcinom dojke), MDA-MB-231 (adenokarcinom dojke), kao i prema EA.hy926 ćelijama (transformisane humane endotelijalne ćelije umbilikalne vene)...Bioactive constituents of plants are in the center of attention of modern cancer research due to their prospective role in cancer chemoprevention based on the suppression of different stages in malignant transformation. The anticancer potential of plant compounds could be attributed to their ability to scavenge free radicals, regulate carcinogen-activating and –detoxifying enzymes and inhibit inflammatory cytokines, than to induce changes in the regulation of growth factors and target molecules in oncogenic signal transduction pathways implicated in cell growth, proliferation, apoptosis, as well as in angiogenesis, invasion and metastasis of cancer cells. The aim of this research was to investigate the cytotoxicity, more exactly to determine the intensity and mechanisms of the cytotoxic actions of the five extracts isolated as fractions from the endemic plant species Helichrysum zivojinii Černjavski et Soška against specific malignant cell type, as well as to determine the selectivity in their antitumor actions against malignant cell lines of different tumor origin and against healthy peripheral blood mononuclear cells (PBMC). To fully evaluate the antitumor potential of extracts, the examination of their effects on the invasiveness of human metastatic malignant cells as well as on the angiogenesis of endothelial cells was done. Chemical characterization of plant extracts was performed in order to understand the relation between qualitative composition and intensity of cytotoxic action of the investigated extracts. The cytotoxic activities of the five extracts of the endemic plant species Helichrysum zivojinii Černjavski et Soška were tested against selected human malignant cell lines: HeLa (cervix adenocarcinoma), Fem-x (melanoma), K562 (myelogenous leukemia), MDA-MB-361 (breast adenocarcinoma), MDA-MB-231 (breast adenocarcinoma), as well as against transformed human umbilical vein endothelial EA.hy926 cells..

Antiproliferative and antibacterial activity of some glutarimide derivatives

Antiproliferative and antibacterial activities of nine glutarimide derivatives (1–9) were reported. Cytotoxicity of compounds was tested toward three human cancer cell lines, HeLa, K562 and MDA-MB-453 by MTT assay. Compound 7 (2-benzyl-2-azaspiro[5.11]heptadecane-1,3,7-trione), containing 12-membered ketone ring, was found to be the most potent toward all tested cell lines (IC50 = 9–27 μM). Preliminary screening of antibacterial activity by a disk diffusion method showed that Gram-positive bacteria were more susceptible to the tested compounds than Gram-negative bacteria. Minimum inhibitory concentration (MIC) determined by a broth microdilution method confirmed that compounds 1, 2, 4, 6–8 and 9 inhibited the growth of all tested Gram-positive and some of the Gram-negative bacteria. The best antibacterial potential was achieved with compound 9 (ethyl 4-(1-benzyl-2,6-dioxopiperidin-3-yl)butanoate) against Bacillus cereus (MIC 0.625 mg/mL; 1.97 × 10−3 mol/L). Distinction between more and less active/inactive compounds was assessed from the pharmacophoric patterns obtained by molecular interaction fields

Synthesis and preliminary screening for the biological activity of some steroidal Δ4-unsaturated semicarbazone derivatives

Eleven new steroidal mono- and bis(semicarbazones)2a–e, 4d and 3a–e have been prepared starting from various 3-oxo-α,β-unsaturated steroids. Mono-semicarbazones 2a–e were further subjected to ethyl chloroacetate in boiling absolute ethanol but, instead of expected intramolecular cyclocondensation reaction products, the new carbazate esters 5a-e were obtained. The structures of all synthesized compounds and identification of each E/Z isomer were deduced by elemental analysis, HRMS, NMR, and IR spectroscopy. Preliminary screening for the cytotoxic activity in vitro of the new compounds has been conducted against three cancer cell lines, K562, Jurkat and HeLa cells. HeLa cells were the most sensitive while K562 cells were the least sensitive to the cytotoxic action of the novel steroid derivatives. Compounds 2e, 3c and 5e were found to have the best but still moderate cytotoxic effects. All tested compounds showed very weak antimicrobial activities. These results demonstrate that the replacement of thioxo group with carbonyl group in steroidal hydrazone derivatives resulted in decrease in their biological activity.Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3083]This is peer-reviewed version of the following article: Živković, M. B.; Novaković, I. T.; Matić, I. Z.; Sladić, D. M.; Krstić, N. M. Synthesis and Preliminary Screening for the Biological Activity of Some Steroidal Δ4-Unsaturated Semicarbazone Derivatives. Steroids 2019, 148, 36–46. [https://doi.org/10.1016/j.steroids.2019.04.010

Hypericum perforatum L. extracts exert cytotoxic effects and show different miRNA signatures in PC-3 and DU 145 prostate cancer cells

Phytochemicals and bioactive substances derived from a wide range of plant extracts have been reported to exert various anticancer effects. Prostate cancer is one of the leading causes of cancer-related deaths within the male population. Prostate cancer-specific miRNA signatures were associated with cancer formation and progression, with various subtypes, and response to therapy. MicroRNA levels of expression were shown to change after the treatment of various compounds and substances extracted from natural products. Natural herbal compounds were shown to induce variations in miRNA expression levels in cancer cells. The aims of this study were to investigate the cytotoxic effects of methanol, ethyl-acetate, and hexane extracts obtained from branch-body part and flowers of Hypericum perforatum L. against humane PC-3 and DU 145 and to test potential miRNA-128/133b/155/193a/206/21/335 signature changes and differences between the two prostate cancer cell lines. Cytotoxic activity of H. perforatum extracts, their effects on cell cycle distribution, and miRNA expression levels were examined in humane PC-3 and DU 145 prostate cancer cells by MTT cell survival assay, flow cytometry, and quantitative real-time PCR. Hexane extract of flowers showed the strongest intensity of cytotoxic activity against PC-3 and DU 145 cells. The highest increase in the percentage of PC-3 cells in the subG1 phase was observed in cell samples treated with hexane extract of flowers and branch-body part. Significant differences in miRNA-128/133b/155/193a/206/21/335 levels were observed between PC-3 and DU 145 cell lines, especially in samples treated with flower extracts compared with the branch-body part. Conclusions: Investigated extracts have significant anticancer potential not only from the aspects of cytotoxicity and cell cycle effects but also from the aspect of lowering oncogenic or increasing tumor-suppressive miRNAs. The best effect might be the increase of tumor-suppressive miR-128 (accompanied by miR-193a) induced by the hexane extract of the flowers, which also exerted the highest cytotoxic activity. Hexane extract of flowers may be the candidate for further investigation for improving the efficiency of standard therapies for PCa. A miRNA signature might be cell-type specific after the treatment with H. perforatum extracts

Supplementary data for article: Živković, M. B.; Novaković, I. T.; Matić, I. Z.; Sladić, D. M.; Krstić, N. M. Synthesis and Preliminary Screening for the Biological Activity of Some Steroidal Δ4-Unsaturated Semicarbazone Derivatives. Steroids 2019, 148, 36–46. https://doi.org/10.1016/j.steroids.2019.04.010

Supplementary material for: [https://www.sciencedirect.com/science/article/pii/S0039128X19300893?via%3Dihub#gp005]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/3082]Related to accepted version: [http://cherry.chem.bg.ac.rs/handle/123456789/3084

Supplementary data for article: Krstić, N. M.; Pavlović, V. D.; Novaković, I. T.; Matić, I. Z.; Sladić, D. Synthesis, Characterization and Biological Evaluation of Some Novel P-Heterocyclic Androst-4-Ene Derivatives. Molecular Diversity 2013, 17 (3), 547–561. https://doi.org/10.1007/s11030-013-9455-9

Supplementary material for: [https://doi.org/10.1007/s11030-013-9455-9]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1376

New Values of Teucrium species: in Vitro Study of Cytotoxic Activities of Secondary Metabolites

The cytotoxicity of seven Teucrium species, a long time ago used as a food spices, for beverages and teas preparing, as well as therapeutics for digestive and respiratory diseases, were examined against human cervix adenocarcinoma HeLa, human melanoma Fem-x, human chronic myelogenous leukemia K562 and human breast adenocarcinoma MDA-MB-361 cells. MTT assay was used for determination of target cell survival. The most prominent cytotoxic effect was observed against K562 cells, especially by T. scordioides, T. montanum and T. botrys. All Teucrium extracts showed good cytotoxic activity on HeLa cells, but very low cytotoxic effect on MDA-MB-361 cells. In addition, the cytotoxic activities of T. scordioides and T. montanum extract were tested on healthy resting and phytohaemagglutinin-stimulated peripheral blood mononuclear cells (PHA-stimulated PBMC). T. scordioides and T. montanum extracts at concentration of 200 µg/ml reduced the resting PBMC and PHA-stimulated PBMC survival up to 10% and 20%, while the reduction of K562 cell survival at the same concentration of extracts was 94% and 97%, respectively. These results point to selectivity in their antitumor actions. Teucrium species can be regarded as promising candidates for natural plant sources of effective biological compounds as a supplements in the food industry, as well as for therapeutic use

Celiac disease-specific and inflammatory bowel disease-related antibodies in patients with recurrent aphthous stomatitis

The etiology of recurrent aphthous stomatitis (RAS) remains unknown. RAS can be presented as primary, idiopathic condition and as a secondary RAS, which is associated with a systemic disease. The aim of our study was to evaluate the presence and concentrations of antibodies specific for celiac disease (CeD) and antibodies related to inflammatory bowel diseases (IBD) in patients with RAS without gastrointestinal symptoms. Antibodies against tissue transglutaminase (anti-tTG), deaminated gliadin peptides (DGP), deaminated gliadinanalogous fragments (anti-GAF-3X) and Saccharomyces cerevisiae (ASCA) were determined by ELISA and anti-neutrophil cytoplasmic antibodies (ANCA) by indirect immunoflurescence (IIF) in 57 patients with RAS and 60 control subjects. The prevalence of CeD specific antibodies did not differ between RAS patients and controls. However, the concentrations of IgA anti-tTG, IgA anti-GAF-3X antibodies in patients with RAS were significantly higher compared to controls (p = 0.002 and p = 0.04 respectively). Histological changes consistent with CeD were confirmed by duodenal biopsy in one RAS patient with highly positive IgA anti-tTG, anti-GAF-3X and anti-DGP antibodies. Higher prevalence along with higher concentrations of IgG ASCA were found in RAS patients compared to controls (p lt 0.01). Patients with positive IgG ASCA in the absence of clinical symptoms decided not to pursue any further testing. Dysfunction of oral mucosa and the exposure to various antigens might be a reason for the loss of tolerance resulting in increased production of autoantibodies. It seems likely that antibodies are markers of aberrant immune response, rather than key effectors involved in the pathogenesis of the disease

Synthesis and antioxidant activity of 1,3,4-oxadiazoles and their diacylhydrazine precursors derived from phenolic acids

Eight 1,3,4-oxadiazole derivatives containing phenolic acid moieties (7a-h) and eight of their diacylhydrazine precursors (6a-h) were synthesized, characterized using spectroscopic methods and examined by scavenging of stable DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals. The most potent phenolic 1,3,4-oxadiazoles showed better DPPH scavenging activity in comparison with their corresponding diacylhydrazine precursors as a result of participation of both aromatic rings and a 1,3,4-oxadiazole moiety in resonance stabilization of the formed phenoxyl radical. Four diacylhydrazines (6d, 6e, 6g, and 6h) and four 1,3,4-oxadiazoles (7d, 7e, 7g and 7h) with the best DPPH scavenging activity, were chosen for further evaluation of their antioxidant potential through various assays. The investigated compounds exerted pronounced ABTS radical scavenging capacity, moderate to good H2O2 scavenging properties and strong ferric ion reducing capacity. Further in vitro evaluation of the antioxidant properties of the most active compounds demonstrated their protective effects in normal lung fibroblasts MRC-5 against hydrogen peroxide induced oxidative stress. Diacylhydrazine 6h increased two times the activity of glutathione peroxidase in treated cells in comparison with a control sample and did not affect the superoxide dismutase activity

Supplementary material for the article: Živković, M. B.; Matić, I. Z.; Rodić, M. V.; Novaković, I. T.; Sladić, D. M.; Krstić, N. M. Synthesis, Characterization and in Vitro Cytotoxic Activities of New Steroidal Thiosemicarbazones and Thiadiazolines. RSC Advances 2016, 6 (41), 34312–34333. https://doi.org/10.1039/c6ra01516f

Supplementary material for: [https://doi.org/10.1039/c6ra01516f]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1913