Ivan Iliev

Synthesis, antiproliferative activity and genotoxicity of novel anthracene-containing aminophosphonates and a new anthracene-derived Schiff base.

A new Schiff base, 9-anthrylidene-furfurylamine and three novel anthracene-containing α-aminophosphonates, [N-methyl(dimethoxyphosphonyl)-1-(9-anthryl)]-p-toluidine, [N-methyl(diethoxyphosphonyl)-1-(9-anthryl)]-p-toluidine and [N-methyl(diethoxyphosphonyl)-1-(9-anthryl)]furfurylamine were synthesized. The compounds have been characterized by elemental analysis, TLC, IR, NMR and fluorescent spectra. The aminophosphonates and their synthetic precursors were tested for in vitro antitumor activity on a panel of seven human epithelial cancer cell lines. Safety testing was performed both in vitro (3T3 NRU test) and in vivo on ICR mice for genotoxicity and antiproliferative activity. 9-Anthrylidene-furfurylamine and [N-methyl(diethoxyphosphonyl)-1-(9-anthryl)]furfurylamine were most potent cytotoxic agents towards colon carcinoma cell line HT-29. The latter compound exhibited also antiproliferative activity to HBL-100, MDA-MB-231 and 647-V cells. The aminophosphonate [N-methyl(dimethoxyphosphonyl)-1-(9-anthryl)]-p-toluidine and its synthetic precursor 9-anthrylidene-p-toluidine were found to be cytotoxic to HBL-100 and HT-29 tumor cell lines, respectively. Moderate genotoxic and antiproliferative activity in vivo and low toxicity to Balb/c 3T3 (clone 31) mouse embryo cells were observed for all tested compounds. The subcellular distribution of two tested compounds in a tumor cell culture system was also studied.

Sodium butyrate enhances the cytotoxic effect of cisplatin by abrogating the cisplatin imposed cell cycle arrest

BackgroundHistone deacetylase inhibitors have been proposed as potential enhancers of the cytotoxic effect of cisplatin and other anticancer drugs. Their application would permit the use of lower therapeutic doses and reduction of the adverse side effects of the drugs. However, the molecular mechanisms by which they sensitize the cells towards anticancer drugs are not known in details, which is an obstacle in developing effective therapeutic protocols.ResultsIn the present work, we studied the molecular mechanisms by which sodium butyrate sensitizes cancer cells towards cisplatin. HeLa cells were treated with 5 mM butyrate, with 8 μM cis-diaminedichloroplatinum II (cisplatin), or with both. Cells treated with both agents showed approximately two-fold increase of the mortality rate in comparison with cells treated with cisplatin only. Accordingly, the life span of albino mice transfected with Ehrlich ascites tumor was prolonged almost two-fold by treatment with cisplatin and butyrate in comparison with cisplatin alone. This showed that the observed synergism of cisplatin and butyrate was not limited to specific cell lines or in vitro protocols, but was also expressed in vivo during the process of tumor development. DNA labeling and fluorescence activated cell sorting experiments showed that cisplatin treatment inhibited DNA synthesis and arrested HeLa cells at the G1/S transition and early S phase of the cell cycle. Western blotting and chromatin immunoprecipitation revealed that this effect was accompanied with a decrease of histone H4 acetylation levels. Butyrate treatment initially reversed the effect of cisplatin by increasing the levels of histone H4 acetylation in euchromatin regions responsible for the G1/S phase transition and initiation of DNA synthesis. This abrogated the cisplatin imposed cell cycle arrest and the cells traversed S phase with damaged DNA. However, this effect was transient and continued only a few hours. The long-term effect of butyrate was a massive histone acetylation in both eu- and heterochromatin, inhibition of DNA replication and apoptosis.ConclusionThe study presents evidence that cell sensitization towards cisplatin by sodium butyrate is due to hyperacetylation of histone H4 in specific chromatin regions, which temporarily abrogates the cisplatin imposed cell cycle arrest.

Chemical composition and antifungal activity of essential oil of Hyssopus officinalis L. from Bulgaria against clinical isolates of Candida species

The chemical composition of hyssop oil from Bulgaria was determined by gas chromatography with flame ionization detection and gas chromatography–mass spectrometry on two different chromatographic columns. The quantity of identified compounds was shown correspond to 97.2% and 98% of the total oil content. Among the detected compounds, cis-pinocamphone (48.98%–50.77%), β-pinene (13.38%–13.54%), trans-pinocamphone (5.78%–5.94%) and β-phellandrene (4.44%–5.17%) were the major compounds. Hyssop oil demonstrated antifungal activity against 52 clinical isolates and reference strains of Candida albicans, Candida glabrata, Candida tropicalis, Candida parapsilosis and Candida krusei. The essential oil characterized with stronger antifungal activity in comparison with pure cis- and trans-pinocamphone, α- and β-pinene and β-phellandrene. Essential oil of Hyssopus officinalis L. from Bulgaria inhibited both fluconazol-sensitive and fluconazol-resistant strains.

Synthesis, characterization, antitumor activity and safety testing of novel polyphosphoesters bearing anthracene-derived aminophosphonate units

Novel polyphosphoesters containing anthracene-derived aminophosphonate units, poly(oxyethylene aminophosphonate)s (4 and 5) and poly[oxyethylene(aminophosphonate-co-H-phosphonate)]s (6 and 7), were synthesized via an addition of poly(oxyethylene H-phosphonate)s to 9-anthrylidene-p-toluidine. The IR, NMR ((1)H, (13)C and (31)P) and fluorescence emission spectral data of the polymers are presented. The copolymers 6 and 7 were tested for in vitro antitumor activity on a panel of seven human epithelial cancer cell lines. Safety testing was performed both in vitro (3T3 NRU test) and in vivo on ICR mice for genotoxicity and antiproliferative activity. The copolymer 7 showed excellent antiproliferative activity to HBL-100, MDA-MB-231, MCF-7 and HepG2 cell lines. However, the in vitro safety testing revealed significant toxicity to Balb/c 3T3 mouse embryo cells. In contrast, the copolymer 6 showed complete absence of cytotoxicity to Balb/c 3T3 cells, but inhibited the growth of breast cancer cells, cervical carcinoma cells (HeLa) and hepatocellular carcinoma cell cultures after prolonged (72h) exposure. The polymers (4-6) exhibited low (4 and 6) to moderate (5) clastogenicity in vivo and slightly inhibited bone marrow cell division, compared to Mitomycin C. The subcellular distribution of the copolymers 6 and 7 were studied in model cell culture systems. The tested polyphosphoesters are expected to act in vivo as prodrugs of aminophosphonates and could be valuable as a new class of biodegradable polymer drug carriers.

Anthracene-Derived Bis-Aminophosphonates: Crystal Structure, In Vitro Antitumor Activity, and Genotoxicity In Vivo

Abstract The X-ray crystal structures of the anthracene-derived bis-aminophosphonates 4.4′-bis[N-methyl(diethoxyphosphonyl)-1-(9-anthryl)]diaminodiphenylmethane (1) and 1,3-bis [N-methyl(diethoxyphosphonyl)-1-(9-anthryl)]diaminobenzene (3) are reported. The X-ray analyses demonstrated that both compounds crystallize in a centrosymmetric manner containing a meso-form (1) and a pair of enantiomers (3). The cytotoxic potential, genotoxicity, and antiproliferative activity of bis-aminophosphonates 1 and bis[N-methyl(diethoxyphosphonyl)-1-(9-anthryl)]benzidine (2), as well as their subcellular distribution in a tumor cell culture system, are also discussed. Compounds 1 and 2 showed optimal antiproliferative activity to human tumor cells from colon carcinoma line HT-29. In vitro and in vivo safety testing revealed that the compounds exert lower toxicity to normal cells as compared with well-known anticancer and cytotoxic agents. Supplemental materials are available for this article. Go to the publishers online edition ofPhosphorus, Sulfur, and Silicon and the Related Elementsto view the free supplemental file. GRAPHICAL ABSTRACT

Biosorption of Pb(II) ions from aqueous solutions by waste biomass of Streptomyces fradiae pretreated with NaOH

Biosorption of Pb(II) ions from a model solution was investigated using Streptomyces fradiae biomass as biosorbent pretreated with sodium hydroxide. The mycelium is a waste product from the biotechnological production of the macrolide antibiotic tylosin in the pharmaceutical industry. The biosorption study was conducted in a batch system with respect to initial pH, initial metal concentration and contact time. For a description of the biosorption equilibrium, Langmuir and Freundlich adsorption models were used. Equilibrium data fitted better to the Langmuir model and the calculated maximum biosorption capacity was 138.88 mg·g−1 at initial pH 5.0, contact time of 120 min, biosorbent dose of 1 g·dm−3 and concentration range for the Pb(II) ions from 10 to 200 mg·dm−3. Pseudo-first and pseudo-second order kinetic models were applied to the experimental data. The results indicated that the Pb(II) uptake process followed the Ho equation. The interference of co-present ions Cu(II) and Zn(II) on the Pb(II) biosorption was also studied. It was determined that at the highest Pb(II) concentration (200 mg·dm−3) Cu(II) and Zn(II) caused 27.22% and 24.88% decreasing in Pb(II) uptake, respectively. The obtained results could be useful in prospective applications of chemically modified waste mycelium of S. fradiae as an alternative biosorbent for Pb(II) removal from aqueous solutions.

Immunological Research on the Protective Properties of a Conjugate of Total Larval Antigen with Hemocyanin Derived from Helix Vulgaris Against Infection with Trichinella Spiralis

ABSTRACT Investigations suggest that resistance against Trichinella spiralis infection can be modified by application of a separate vaccine preparation, in this case—conjugated total antigen from Trichinella spiralis larvae with hemocyanin from Helix vulgaris. These effects are generally to reduce resistance at the level of the gut, and enhance it against the systemic phase of the infection. The mechanism, which induces these alterations is still not completely clear. Our results showed reduced tissue penetration of the nematode worms in muscles. In addition we have observed solid protective humoral and cellular immune response.

Suboptimal growth temperatures enhance the biological activity of cultured cyanobacterium Gloeocapsa sp.

The cytotoxic, antibacterial, and antifungal activities of cyanobacterium Gloeocapsa sp. strain Gacheva 2007/R-06/1 were investigated and the possibility for an enhancement of these activities by changing the culture conditions evaluated. Fatty acids of this cyanobacterium were found to be active against Streptococcus pyogenes. Exopolysaccharides inhibited the growth of both Gram-positive and Gram-negative bacteria and the fungus Candida albicans. Both exopolysaccharides and fatty acid mixtures also significantly decreased the viability of human cervical carcinoma cells, HeLa. Greater biological activities were exhibited by Gloeocapsa sp., cultured at suboptimal temperatures (15–26°C) than at optimal and supraoptimal ones. In comparison with higher light intensity, the low-light cultivation stimulated the cytotoxicity of the fatty acids. In general, low temperatures decreased the growth of Gloeocapsa sp., but promoted its biological activity. Prolonged cultivation also had a beneficial impact on the bioactivity. Compared to 4xa0days, the 17-day cultivation resulted in fourfold higher antibacterial and antifungal activities of exopolysaccharides and more than twice increases in their cytotoxicity. The study revealed that this cyanobacterial isolate is a new source of natural products with potential for pharmacological and medical applications.

Antibiotic resistance of Gram-negative benthic bacteria isolated from the sediments of Kardzhali Dam (Bulgaria)

The aim of the present study was to carry out a preliminary assessment for the occurrence of bacterial strains resistant to frequently used antibiotics in the sediments beneath the sturgeon cage farm in Kardzhali Dam (Bulgaria). Samples were taken from the top 2 cm of sediments under a fish farm and from a control station in the aquatory of the reservoir in the period July–October 2011. Surveillance of bacterial susceptibility to 16 antimicrobial agents was performed for 160 Gram-negative strains (Pseudomonas mandelii – 100 strains; Hafnia alvei – 30 strains; and Raoultella ornithinolytica – 30 strains). No significant differences in the resistance to the tested antibiotics were observed between the strains isolated from the two stations (analysis of variance, P > 0.05). Widespread resistance to penicillins and certain cephalosporin antibiotics was observed in both stations. None of the studied strains showed resistance to the aminoglycoside antibiotics gentamicin and amikacin, or to ciprofloxacin. Minimal Inhibitory Concentrations (MIC) were determined for five of the tested antimicrobial agents by the microdilution antibiotic sensitivity assay. The data indicate that amikacin, tetracycline and ciprofloxacin effectively suppress the growth of the tested micro-organisms. The isolates from genus Pseudomonas showed the highest MIC and were characterized by the highest percentage of antibiotic resistance.

In Vitro Assessment of the Cytotoxic Effects of Sulfo-Arginine Analogues and their Hydrazide Derivatives in 3T3 and HepG2 Cells

ABSTRACT The cytotoxic activities of sulfo-arginine analogues sArg, NsArg, and their hydrazide derivatives, sArg-CONHNH2, sArg-CONHNC6H5, sArg-CONHN(CH2CH2Cl)2, NsArg-CONHNH2, NsArg-CONHNC6H5, NsArg-CONHN(CH2CH2Cl)2, on 3T3 and HepG2 cells were examined. The substitution in the carboxylic group of sArg increases the cell growth inhibitory effects of the compounds, especially in the case of the bis-(2-chloroethyl)hydrazide substitute. Similar correlation was observed in the case of NsArg and its analogues, but here the most pronounced effect was observed with the analogue NsArg-CONHNH2.