Antitumor potential of new steroidal hydrazone derivatives

Kako maligne ćelije vremenom mutiraju i postaju rezistentne na postojeće lekove postoji stalna potreba za novim antitumorskim lekovima. Steroidni molekuli igraju vrlo važne uloge u biološkim sistemima. Usled hidrofobnosti nesmetano prolaze kroz ćelijske membrane pa su pogodan polazni materijal za dizajn novih lekova. Trasformacijama krutog tetracikličnog steroidnog skeleta nastaju biološki aktivni molekuli sa novim fiziološkim funkcijama. Sa druge strane, hidrazoni sadrže heteroatome i smatraju se značajnim intermedijerima pri sintezi heterocikličnih jedinjenja. Stoga je sintetisano i okarakterisano pedeset novih derivata steroidnih hidrazona, a zatim je određena njihova citotoksičnost i ispitani su mehanizmi citotoksičnog delovanja. Prema testiranim malignim ćelijskim linijama (HeLa, K562, MDA-MB-361, MDA-MB-453, LS174, A549), naročito prema HeLa ćelijama adenokarcinoma cerviksa i prema K562 ćelijama hronične mijeloidne leukemije, nova jedinjenja su ispoljila selektivnu citotoksičnost zavisnu od koncentracije na mikrogramskoj skali, u mnogim slučajevima bolju od hemioterapeutika cisplatina. Nasuprot tome, pokazala su vrlo slabu citotoksičnost na humane PBMC, što se ogleda u koeficijentima selektivnosti daleko višim od vrednosti 2,5. Morfološkom analizom HeLa ćelija tretiranih novim derivatima utvrđeno je njihovo pro-apoptotsko dejstvo što je potvrđeno rezultatima protočne citometrije. Određivanjem ciljnih kaspaza ustanovljeno je da do apoptoze dolazi posredstvom kaspaze-3, -8 i -9. Svi ispitivani derivati su pokazali sposobnost da inhibiraju angiogenezu ćelija EA.hy926 in vitro. Testirani tiazolidin-4-oni nisu prouzrokovali značajne promene u nivoima ekspresije gena MMP2, MMP9 i VEGFA u tretiranim HeLa ćelijama. Rezultati sprovedenih in vitro istraživanja mogu da ukažu na značajan antitumorski potencijal što ova jedinjenja ubraja u potencijalne kandidate za in vivo ispitivanja

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

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 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

Synthesis, characterization and biological activity of Pt(II) complexes with steroidal thiosemicarbazones

In this work, Pt(II) complexes of previously synthesized steroidal thiosemicarbazones were synthesized and characterized. The ligands and their metal complexes were studied by analytical and spectroscopic data (elemental analysis, IR, 1D-NMR and 2D-NMR, HSQC, HMBC, NOESY, COSY), the analysis of which enabled complete 1H and 13C assignments of each compound including E and Z isomers. All the synthesized ligands and complexes were screened for their cytotoxic and antimicrobial activity. The results demonstrate that the new steroidal thiosemicarbazone complexes were significantly less cytotoxic than the corresponding steroidal thiosemicarbazones. In addition, complexes showed lower antimicrobial activity than the standard drugs, similar to the activity of the starting thiosemicarbazones

Ekstrakt ploda morača kao potencijalni prirodni aditiv u prehrambenoj industriji

In this study, the polyphenol profile and antioxidant activity of the hydro-ethanolic extract of the fennel fruit were examined in order to investigate the possibility of its application as a potential functional food additive. Total phenols were analyzed by the method of Folin-Ciocalteu, while total flavonoids were determined by the aluminum chloride colorimetric method. The separation and quantification of phenolic compounds were performed by LC-MS/MS analysis, using a multiple reaction monitoring (MRM) mode. The antioxidant capacity was determined by FRAP and DPPH assays. The high values of total phenolics and flavonoids were found, as well as high antioxidant activity which amounted to 9023.33 ± 38.19 µmol Fe(II)/l and 3.73 ± 0.04 mmol TE/l, tested by FRAP and DPPH assays, respectively. Among the identified phenolic compounds, p-hydroxybenzoic and chlorogenic acids were detected as predominant. The obtained results indicated that the hydro-ethanolic extract of the fennel fruit can be used in food industry as a potential natural antioxidant.U ovom radu određivan je sadržaj polifenola i antioksidativna aktivnost vodeno-etanolnog ekstrakta morača, sa ciljem ispitivanja mogućnosti njegove primene kao potencijalnog funkcionalnog aditiva. Ukupni fenoli su analizirani metodom po Folin-Ciocalteu, dok je ukupan sadržaj flavonoida određen kolorimetrijskom metodom primenom aluminijum hlorida. Razdvajanje i kvantifikacija fenolnih jedinjenja postignuti su upotrebom LC-MS/MS metode u režimu koji omogućava istovremeno praćenje više jonskih prelaza. Antioksidativni kapacitet je određivan primenom testova FRAP i DPPH. U testiranom ekstraktu dobijene su visoke vrednosti za ukupne fenole i flavonoide, a dobijena je i visoka vrednost antioksidativne aktivnosti, koja je iznosila 9023.33 ± 38,19 mmol Fe(II)/l i 3,73 ± 0,04 mmol TE/l, računato primenom testa FRAP odnosno testa DPPH. Među fenolnim jedinjenjima, phidroksibenzoeva i hlorogena kiselina su pronađene kao dominantne. Dobijeni rezultati ukazuju na to da se ekstrakt morača može primenjivati u prehrambenoj industriji kao potencijalni prirodni antioksidans

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

Development and validation of an LC-MS/MS method with a multiple reactions monitoring mode for the quantification of vanillin and syringaldehyde in plum brandies

An ultra-performance liquid chromatographic triple quadrupole mass spectrometric (UPLC-QqQ-MS/MS) method with a multiple reactions monitoring mode (MRM) was developed and validated for the quantification of vanillin and syringaldehyde in plum brandy. The method showed good linearity (0.05 to 10 mg L-1) and low limits of detection and quantification (the LOD and LOQ values were 11.6 and 38.2 mu g L-1 for vanillin, and 12.7 and 42.0 mu g L-1 for syringaldehyde, respectively). The overall intra-day and inter-day variations were less than 4.21 % and the overall recovery was over 93.0 %. The correlation coefficients (R-2) of the calibration curves were higher than 0.9999. In order to evaluate whether the method was suitable for use as a routine analytical tool, vanillin and syringaldehyde were determined in 31 samples of Serbian plum brandy

Supplementary material for the article: Živković, M. B.; Matić, I. Z.; Rodić, M. V.; Novaković, I. T.; Krivokuća, A. M.; Sladić, D. M.; Krstić, N. M. Anticancer Potential of New Steroidal Thiazolidin-4-One Derivatives. Mechanisms of Cytotoxic Action and Effects on Angiogenesis in Vitro. Journal of Steroid Biochemistry and Molecular Biology 2017, 174, 72–85. https://doi.org/10.1016/j.jsbmb.2017.07.031

Supplementary material for: [https://doi.org/10.1016/j.jsbmb.2017.07.031]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2550]Related to accepted version: [http://cherry.chem.bg.ac.rs/handle/123456789/3191

Histological evaluation of direct pulp capping with novel nanostructural materials based on active silicate cements and Biodentine® on pulp tissue

The aim of this study was to examine the effect of Biodentine® and two new nanostructured materials based on active silicate cements on exposed tooth pulp of Vietnamese pigs. The study comprised 40 teeth in two Vietnamese pigs (24 months old). After class V cavity preparation, the pulp on each tooth was exposed using a small round bur. The following materials were applied on pulp exposures: Biodentine® (10 teeth), ALBO MPCA-I (10 teeth), and ALBO MPCA-II (10 teeth). In the control group, exposed pulp was covered with ProRoot MTA® (10 teeth). After the observation period of 28 days, the animals were sacrificed and the teeth prepared for histological analysis. Light microscope was used for the analysis of dentin bridge formation, tissue reorganization and inflammation, and the presence of bacteria in the pulp. In the group of Biodentine®, a complete dentin bridge was noted in 3 cases, while incomplete dentin bridge in the form of dental islets was detected in 4 cases. Nanostructured material ALBO-MPCA I provided complete dentin bridge formation in 5 teeth, in 3 teeth the formed dentin bridge was incomplete. ALBO MPCA-II showed complete closure of the pulp opening by dentin bridge in 4 samples, while in the same number of teeth it was incomplete. In the control group, 4 teeth showed a complete dentin bridge, whereas in 6 teeth it was incomplete. Histological analysis indicated favourable therapeutic effects of Biodentine® and the two materials ALBO-MPCA I and ALBO-MPCA II after teeth pulp capping in Vietnamese pigs. Pulp reaction was similar to that caused by ProRoot MTA®. [Projekat Ministarstva nauke Republike Srbije, br. ON172026