Anticholinesterase and Serotoninergic Evaluation of Benzimidazole-Carboxamides as Potential Multifunctional Agents for the Treatment of Alzheimer's Disease.

The etiology and pathogenesis of Alzheimer's disease are multifactorial, so one of the treatment strategies is the development of the drugs that affect several targets associated with the pathogenesis of the disease. Within this roadmap, we investigated the interaction of several substituted 1,3-dihydro-2-oxo-1H-benzimidazol-2-ones with their potential molecular targets: cholinesterases (ChE) and three types of the Gs-protein-coupled serotonin receptors (5-HTR) 5-HT6, 5-HT4 and 5-HT7 (5-HT4R, 5-HT6R and 5-HT7R, respectively). A microplate modification of the Ellman method was used for the biochemical analysis of the inhibitory ability of the drugs towards ChE. Molecular modeling methods, such as molecular docking and molecular dynamics (MD) simulation in water and the lipid bilayer, were used to study the interaction of the compounds with ChE and 5-HTR. In vitro experiments showed that the tested compounds had moderate anticholinesterase activity. With the help of molecular modeling methods, the mechanism of interaction of the tested compounds with ChE was investigated, the binding sites were described and the structural features of the drugs that determine the strength of their anticholinesterase activity were revealed. Primary in silico evaluation showed that benzimidazole-carboxamides effectively bind to 5-HT4R and 5-HT7R. The pool of the obtained data allows us to choose N-[2-(diethylamino)ethyl]-2-oxo-3-(tert-butyl)-2,3-dihydro-1H-benzimidazole-1-carboxamide hydrochloride (compound 13) as the most promising for further experimental development.

Natural Phaeosphaeride A Derivatives Overcome Drug Resistance of Tumor Cells and Modulate Signaling Pathways.

In the present study, natural phaeosphaeride A (PPA) derivatives are synthesized. Anti-tumor studies are carried out on the PC3, K562, HCT-116, THP-1, MCF-7, A549, NCI-H929, Jurkat, and RPMI8226 tumor cell lines, and on the human embryonic kidney (HEK293) cell line. All the compounds synthesized turned out to have better efficacy than PPA towards the tumor cell lines listed. Among them, three compounds exhibited an ability to overcome the drug resistance of tumor cells associated with the overexpression of the P-glycoprotein by modulating the work of this transporter. Luminex xMAP technology was used to assess the effect of five synthesized compounds on the activation of intracellular kinase cascades in A431 cells. MILLIPLEX MAP Multi-Pathway Magnetic Bead 9-Plex was used, which allowed for the simultaneous detection of the following nine phosphorylated protein markers of the main intracellular signaling pathways: a universal transcription factor that controls the expression of immune-response genes, apoptosis and cell cycle NFκB (pS536); cAMP-dependent transcription factor (CREB (pS133); mitogen-activated kinase p38 (pT180/pY182); stress-activated protein kinase JNK (pT183/pY185); ribosomal SK; transcription factors STAT3 (pS727) and STAT5A/B (pY694/699); protein kinase B (Akt) (pS473); and kinase regulated by extracellular signals ERK1/2 (pT185/pY187). The effect of various concentrations of PPA derivatives on the cell culture was studied using xCelligence RTCA equipment. The compounds were found to modulate JNK, ERK1/2, and p38 signaling pathways. The set of activated kinase cascades suggests that oxidative stress is the main probable mechanism of the toxic action of PPA derivatives.

Nutrient and metal pollution of the eastern Gulf of Finland coastline: Sediments, macroalgae, microbiota.

The anthropogenic pollution along the coastline of the eastern Gulf of Finland was studied through a range of methods, including analyses of metal contamination in water, surface sediments, accumulated algal biomass and its correlation with resistant microbiota. According to concentrations, the main pollutants in water were copper and manganese. Influence of Nuclear Power Plant was remarkable in adjacent areas and was expressed in high concentrations of molybdenum, nickel, copper and other elements in the water. Relatively high concentrations of copper, lead and zinc were found in sediments. Microbial tolerance appeared to be correlated with the concentration of the metals in sediments. Higher tolerance levels were found in sediment samples from more polluted stations. Macroalgae, which were massively developed in the coastal zone, had shown high level of metal bioaccumulation. Analyses of carbon, nitrogen and phosphorus content of algal tissues allowed the estimation of additional nutrient loading from accumulated decaying algal biomass on the coastal zone of the eastern Gulf of Finland. Mass development of algae in coastal area may contribute to accumulation of organic matter and associated metals. In our study the highest metal concentrations in sediments were found at the sites with dense and continuous layer of fresh and decaying macroalgal biomass, accompanied by hypoxic conditions. Also our study has shown that accumulated biomass may be a significant source of nutrients in the coastal ecosystem.

Biological evaluation and determination of the absolute configuration of chloromonilicin, a strong antimicrobial metabolite isolated from Alternaria sonchi.

Chloromonilicin was isolated for the first time from Alternaria sonchi, a mycoherbicide proposed for the control of the noxious weed Sonchus arvensis. The already known alternethanoxins A and B and the three recently isolated phytotoxic polycyclic ethanones named alternethanoxins C-E were also isolated from the same source. Chloromonilicin was identified by spectroscopic data (essentially one-dimensional NMR, 2-dimensional NMR and high-resolution ESI-MS) and its structure was confirmed by single X-ray analysis, which also allowed the assignment of the absolute configuration. This latter was independently confirmed by electronic CD calculations. When chloromonilicin was tested for its antimicrobial activity, it was active at concentrations 0.5-1 µg per disc against four bacterial species and a yeast fungus. The compound inhibited conidial germination of four plant pathogens at concentration of 1-10 µg ml(-1). No phytotoxic activity of this antibiotic by leaf-disc puncture bioassay was detected.

Alternethanoxins C-E, Further Polycyclic Ethanones Produced by Alternaria sonchi, a Potential Mycoherbicide for Sonchus arvensis Biocontrol.

Three new polycyclic ethanones, named alternethanoxins C-E, were isolated together with the well-known and closely related alternethanoxins A and B, from the solid culture of Alternaria sonchi, a fungal pathogen proposed for perennial sowthistle (Sonchus arvensis L.) biocontrol. Alternethanoxins C-E were characterized by spectroscopic methods (essentially NMR and HRESI MS) as 2'-dihydroxymethyl-2,5,6,6'-tetrahydroxy-3'-methoxy-biphenyl-3-carboxylic acid methyl ester, 1,4,6,9,10-pentahydroxy-7-methoxy-6H-benzo[c]chromene-2-carboxylic acid methyl ester, and 7,9-dihydroxy-2-methoxy-9H-4,8-dioxa-cyclopenta[def]phenanthrene-5-carboxylic acid methyl ester. When assayed on leaf segments of weeds (Sonchus arvensis and Elytrigia repens), alternethanoxins A and C showed phytotoxic activity inducing notable necrotic lesions. Alternethanoxins C and D possess notable antimicrobial activity when tested against Bacillus subtilis (MIC 10 µg/disc) and Candida tropicalis (MIC 25 µg/disc). Alternethanoxins A and B had low activity against these microbes, while alternethanoxin E was inactive.