Search

Smoking mixtures containing cannabimimetic indoles may still be illegally sold in Russia. Although a method for their analysis is required for forensic toxicology authorities, the detection of synthetic cannabinoids is a complicated analytical task because of low anticipated concentrations in urine and the lack of in vivo data on their metabolism. Here, the urinary metabolites of 1-pentyl-3-(1-naphthoyl)indole (JWH-018) and a procedure for determining them in urine are reported. Using gas and high-performance liquid chromatography combined with tandem mass spectrometry, two main monohydroxylated metabolites were identified in urine. Based on differences in their electron ionization MS/MS spectra, it is supposed that one of them is formed by the hydroxylation of an indole ring and the other, by the hydroxylation of a pentyl side chain. The main metabolites are almost completely excreted as conjugates with glucuronic acid. The structure of minor metabolites was proposed. The parent compound was not detected in urine at a level of 50 pg/mL 12 h after administration. Keywords: JWH-018, synthetic cannabinoids, smoking mixtures, metabolism, GC/MS, HPLC/MS. DOI:10.1134/S1061934811130090, INTRODUCTION In the last two years, so-called smoking mixtures ('Spice' etc.), which contain synthetic cannabinoids, mainly 1-pentyl-3-(1-naphthoyl)indole (JWH-018), appeared on the market of many countries under the guise of fragrances. [...]

Research has been conducted on the antipyretic drug paracetamol. The oxidation chemistry of this drug with UV/H (sub)2 O (sub)2 system has been examined via various techniques including (super)15 N NMR analysis and the details are reported.

The authors discusses the research conducted on the hypoxia-inducible factor (HIF). The topics of interest include HIF control by prolyl hydroxylation, asparaginyl hydroxylation and HIF transactivation function, and therapeutic opportunities.

Research has been conducted on the brown-rot basidiomycetes. The utilization of benzaldehyde as the aromatic substrate for these basidomycetes has been carried out and the results have revealed benzylic oxidation and reduction.

Research has been conducted on the prostaglandins. The route to the F (sub)2t -isoprostanes has been developed via the use of C20 precursor's biomimetic radical cyclization and the results demonstrate the formation of (15R)- and (15S)-F (sub)2t -isoprostane diastereoisomers.

Research has been conducted on the racemates and 2-fluorocycloalkyl N-phenylcarbamates. The biohydroxylation of these compounds with Beauveria bassiana has been investigated and the results indicate the isolation of the diastereoisomeric product mixtures hydroxylated in the 4-position.

Research has been conducted on the pyrrolidine based scaffold comprised of 2,5-dideoxy-2,5-imino-D-idaric acid. The development of this compound has been carried out and the results indicate that the peptide strand attachment to the carboxylic groups has led to the peptidomimetics.

Research has been conducted on the triple-labeled vitamin D and its hydroxylated and dihydroxylated metabolites. The synthesis of these isotope-labeled drug molecules has been carried out via the coupling of [9- (super)13C]-Grundmann's ketone with the labeled A ring enyne fragments and the NMR experiment on a model micellar system is discussed.

Research has been conducted on all-cis 2,3,4,5-substituted pyrrolidines. The enantioselective synthesis of these sterically congested pyrrolidines has been carried out from D- and L-serine and pyrrolidines' all-cis configuration has been established via the X-ray analysis.

Research has been conducted on the bicyclic 2-pyridone dipeptide mimetic. The preparation of this compound in the optically active form and the steps of the synthesis which include osmium-catalyzed asymmetric olefin dihydroxylation are described.

Research has been conducted on the N-benzylpyrrolidine. The hydroxylation of this compound has been carried out with the Sphingomonas sp. HXN-200 resting cells to produce N-benzyl-3-hydroxypyrrolidine and the results indicate that the change of the docking/protecting group in pyrrolidines improves the hydroxylation activity and enantioselectivity of the compound.

Research has been conducted on the fosfomycin, organophosphonic acid isolated from the Streptomyces fradiae fermentation broths. The synthesis of this fosfomycin has been carried out via the asymmetric dihydroxylation reaction of dibenzyl (E)-1-propenylphosphonate with the modified asymmetric dihydroxylation-mix-alpha-(x3) and the results are reported.

Research has been conducted on the cis-1,2-dihydrocatechol obtained by microbial dihydroxylation of bromobenzene. The conversion of this dihydrocatechol into the enantiomer of the cyclolysine-based marine natural product bengamide E is described.

Research has been conducted on the yeast Trichosporon mucoides grown on glucose or phenol. The ability of this yeast to transform biphenyl into mono-, di- and trihydroxylated derivatives hydroxylated on the aromatic rings has been investigated.

The stereoselective synthesis of fluorinated analogues of (2S,3R)-(3,4-dihydroxyphenyl)serine has been carried out. The results of the aldol condensation of 3,4-dibenzyloxy-2-fluorobenxaldehyde and 4,5-dibenzyloxy-2-fluorobenxaldehyde with oxazolidinone are discussed.

Research has been conducted on the Wittig/dihydroxylation reactions. The improved strategy for the higher sugars synthesis by the Wittig/dihydroxylation of unprotected aldoses has been developed and the results are discussed.

Research has been conducted on (-)-wilkstromol, a bioactive alpha-hydroxylated lactone lignan. The synthesis of (-)-wilkstromol from natural malik acid via consecutive alkylation strategy is discussed.

Research has been conducted on the alpha,beta-unsaturated nitriles. Results indicate that these nitriles are readily synthesized by the chelation-controlled ring opening of epoxy, tetrahydrofuranyl and tetrahydropyranyl acetonitriles.

A major catabolic pathway for the gibberellins (GAs) is initiated by 2[Beta]-hydroxylation, a reaction catalyzed by 2-oxoglutarate-dependent dioxygenases. To isolate a GA 2[Beta]-hydroxylase cDNA clone we used functional screening of a cDNA library from developing cotyledons of runner bean (Phaseolus coccineus L.) with a highly sensitive tritium-release assay for enzyme activity. The encoded protein, obtained by heterologous expression in Escherichia coli, converted [GA.sub.9] to [GA.sub.51] (2[Beta]-hydroxy[GA.sub.9]) and [GA.sub.51]-catabolite, the latter produced from [GA.sub.51] by further oxidation at C-2. The enzyme thus is multifunctional and is best described as a GA 2-oxidase. The recombinant enzyme also 2[Beta]-hydroxylated other [C.sub.19]-GAs, although only [GA.sub.9] and [GA.sub.4] were converted to the corresponding catabolites. Three related cDNAs, corresponding to gene sequences present in Arabidopsis thaliana databases, also encoded functional GA 2-oxidases. Transcripts for two of the Arabidopsis genes were abundant in upper stems, flowers, and siliques, but the third transcript was not detected by Northern analysis. Transcript abundance for the two most highly expressed genes was lower in apices of the GA-deficient ga1-2 mutant of Arabidopsis than in wild-type plants and increased after treatment of the mutant with [GA.sub.3]. This up-regulation of GA 2-oxidase gene expression by GA contrasts GA-induced down-regulation of genes encoding the biosynthetic enzymes GA 20-oxidase and GA 3[Beta]-hydroxylase. These mechanisms would serve to maintain the concentrations of biologically active GAs in plant tissues.

Research has been conducted on cis and trans substituted 4-tert-butoxycarbonyl cyclohexylglycines. The synthesis of these cyclohexylglycines has been carried out via vinyl styrene asymmetric aminohydroxylation and the results are described.

Hypoxia-inducible factor-1[Alpha] [(HIF-1[Alpha]).sup.1] is a global transcriptional regulator of the hypoxic response. Under normoxic conditions, HIF-1[Alpha] is recognized by the von Hippel-Lindau tumor-suppressor protein (VHL), a component of an E3 ubiquitin ligase complex. This interaction thereby promotes the rapid degradation of HIF-1[Alpha]. Under hypoxic conditions, HIF-1[Alpha] is stabilized. We have previously shown that VHL binds in a hypoxia-sensitive manner to a 27-aa segment of HIF-1[Alpha], and that this regulation depends on a post-translational modification of HIF-1[Alpha]. Through a combination of in vivo coimmunoprecipitation assays using VHL and a panel of point mutants of HIF-1[Alpha] in this region, as well as MS and in vitro binding assays, we now provide evidence that this modification, which occurs under normoxic conditions, is hydroxylation of Pro-564 of HIF-1[Alpha]. The data furthermore show that this proline hydroxylation is the primary regulator of VHL binding.

To facilitate plant colonization, some pathogenic fungi produce phytotoxic metabolites that damage tissues; plants may be resistant to a particular pathogen if they produce an enzyme(s) that catalyzes detoxification of this metabolite(s). Alternaria blackspot is one of the most damaging and significant fungal diseases of brassica crops, with no source of resistance known within the Brassica species. Destruxin B is the major phytotoxin produced by the blackspot-causing fungus, Alternaria brassicae (Berkley) Saccardo. We have established that a blackspot-resistant species (Sinapis alba) metabolized [sup.14]C-labeled destruxin B to a less toxic product substantially faster than any of the susceptible species. The first metabolite, hydroxydestruxin B ([sup.14]C-labeled), was further biotransformed to the [Beta]-D-glucosyl derivative at a slower rate. The structures of hydroxydestruxin B and [Beta]-D-glucosyl hydroxydestruxin B were deduced from their spectroscopic data [NMR, high resolution (HR)-MS, Fourier transform infrared (FTIR)] and confirmed by total chemical synthesis. Although these hydroxylation and glucosylation reactions occurred in both resistant (S. alba) and susceptible (Brassica napus, Brassica juncea, and Brassica rapa) species, hydroxylation was the rate limiting step in the susceptible species, whereas glucosylation was the rate limiting step in the resistant species. Remarkably, it was observed that the hydroxydestruxin B induced the biosynthesis of phytoalexins in blackspot-resistant species but not in susceptible species. This appears to be a unique example of phytotoxin detoxification and simultaneous phytoalexin elicitation by the detoxification product. Our studies suggest that S. alba can overcome the fungal invader through detoxification of destruxin B coupled with production of phytoalexins.