Enthalpies of Combustion and Formation of Severely Crowded Methyl-Substituted 1,3-dioxanes. The Magnitudes of 2,4- and 4,6-diaxial Me,Me-Interactions and the Chair-2,5-twist Energy Difference.

Enthalpies of combustion of 2,2-trans-4,6- (1) and 4,4,6,6-tetramethyl- (2) and 2,4,4,6,6- (3) and 2,2,4,4,6-pentamethyl-1,3-dioxanes (4) were determined to estimate their enthalpies of formation in the gas phase. By comparing the latter with the corresponding enthalpies estimated based on the various bond-bond interactions allowed to determine the chair-2,5-twist energy difference (ΔHCT = 29.8 kJ mol-1) for 1 since C-13 shift correlations indicate that it escapes to the 2,5-twist form where the 2-methyl groups are isoclinal and 4- and 6-methyl groups pseudoequatorial to avoid syn-axial interactions. Compounds 2 and 3 in turn give the values 21.0 and 21.6 kJ mol-1 for the 4,6-diaxial Me,Me-interaction. Finally compound 4, which retains the chair conformation to avoid pseudoaxial interactions in the twist forms gives the value 19.5 kJ mol-1 for the 2,4-diaxial Me,Me-interaction indicating that its chair form appears to be somewhat deformed.

Structural studies, homology modeling and molecular docking of novel non-competitive antagonists of GluK1/GluK2 receptors.

Non-competitive ligands of kainate receptors have focused significant attention as medicinal compounds because they seem to be better tolerated than competitive antagonists and uncompetitive blocker of these receptors. Here we present structural studies (X-ray structure determination, NMR and MS spectra) of novel indole-derived non-competitive antagonists of GluK1/GluK2 receptors, homology models of GluK1 and GluK2 receptors based on novel AMPA receptor template as well as molecular docking of ligands to their molecular targets. We find that the allosteric site is in the receptor transduction domain, in one receptor subunit, not between the two subunits as it was indicated by our earlier studies.

The pseudo-Michael reaction of 1-aryl-4,5-dihydro-1H-imidazol-2-amines with ethyl ethoxymethylenecyanoacetate.

ABSTRACT: The pseudo-Michael reaction of 1-aryl-4,5-dihydro-1H-imidazol-2-amines with ethyl 2-cyano-3-methoxyprop-2-enoate (ethyl ethoxymethylenecyanoacetate) is investigated. At -10 °C reaction takes place on the exocyclic nitrogen atom, giving exclusively ethyl esters of 2-cyano-3-[(1-phenyl-4,5-dihydro-1H-imidazol-2-yl)amino]prop-2-enoic acid. The formation of isomeric enamines which may be a theoretical product of the reaction on N3 ring nitrogen atom is not observed. The N6 enamines, heated in boiling acetic acid, yield cyclic 1-aryl-5-oxo-2,3-dihydroimidazo[1,2-a]pyrimidine-6-carbonitriles. Heating of the enamines to the temperature of 120-140 °C without a solvent makes it possible to obtain a mixture of 1-aryl-5-oxo-2,3-dihydroimidazo[1,2-a]pyrimidine-6-carbonitriles and ethyl 1-aryl-5-imino-2,3-dihydroimidazo[1,2-a]pyrimidine-6-carboxylates. The reaction of the respective hydrobromides of 1-aryl-4,5-dihydro-1H-imidazol-2-amines with ethyl ethoxymethylenecyanoacetate in the presence of triethylamine gives selectively 1-aryl-5-oxo-1,2,3,5-dihydroimidazo[1,2-a]pyrimidine-6-carbonitriles.

1-Oxo-1,3-dithiolanes--synthesis and stereochemistry.

1-Oxo-1,3-dithiolane (4) and its cis- and trans-2-methyl (5,6), -4-methyl (7,8) and -5-methyl (9,10) derivatives were prepared by oxidizing the corresponding 1,3-dithiolanes (1-3) with NaIO(4) in water. The oxides were purified and their isomers separated using thin layer chromatography. The structural characterization was carried out with (1)H and (13)C NMR spectroscopy and molecular modelling. The sulfoxides 4-6 and 8-10 attain two S(1) type envelopes (sometimes slightly distorted) the S=O(ax) envelope greatly dominating. Cis-4-methyl-1-oxo-1,3-dithiolane is a special case exhibiting both two closely related S=O(ax) (30 and 27%) as well as S=O(eq) (21 and 22%) forms [S(1) and C(4) envelopes, respectively]. The relative energies of these conformations, the values of (1)H-(1)H coupling constants and (1)H and (13)C chemical shifts were estimated by computational methods and they support well the conclusions based on the experimental data.

Polyketide derivatives active against Botrytis cinerea in Gerbera hybrida.

A previously isolated cDNA molecule from Gerbera hybrida (Asteraceae) codes for a new chalcone synthase-like polyketide synthase, 2-pyrone synthase (2PS). 2PS is able to synthesise 4-hydroxy-6-methyl-2-pyrone (triacetolactone), a putative precursor for gerberin and parasorboside, two abundant glucosides in gerbera. In this study, we show that gerbera plants transformed with the gene for 2PS in an antisense orientation and unable to synthesise gerberin and parasorboside are susceptible to Botrytis cinerea infection. In addition to the preformed glucosides, the transgenic plants also lack several compounds that are induced in control plants when infected with the mould. Some of these induced substances are effective in inhibiting fungal growth both in vitro and in vivo. Two of the phytoalexins were identified as the aglycones of gerberin and trans-parasorboside. The third phytoalexin is a rare coumarin, 4-hydroxy-5-methylcoumarin; however, it is typical of many plants of the sunflower family Asteraceae. The coumarin cannot be structurally derived from either gerberin or parasorboside, but may be derived from a related polyketide intermediate.

Phenolic compounds from Eucalyptus gomphocephala with potential cytotoxic and antioxidant activities.

Two new phenolic compounds, 2,4,6-trihydroxy-5-methyl-acetophenone 2-O-beta-D-glucopyranoside (3), and benzyl alcohol 7-O-(3',4',6'-tri-O-galloyl)-beta-D-glucopyranoside (8), together with eight known phenolic compounds, were isolated from the 70% aqueous acetone extract of Eucalyptus gomphocephala DC. (Myrtaceae). The isolated compounds were elucidated based on their 1H, 13C, DQF-COSY, selective 1D-TOCSY, HSQC, and HMBC NMR spectroscopic and ESI-MS data. The antioxidant effect of the phenolic compounds was tested using 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical and super oxide anion radical scavenging assays. The cytotoxicity of the isolated compounds was evaluated using HeLa cell line.

Electron ionization mass spectra and tautomerism of substituted 2-phenacylquinolines.

Tautomerism has been studied conventionally in solutions or in the solid state. However, the importance of mass spectrometry in the gas phase was realized relatively late. 2-Phenacylquinolines are known to undergo ketimine-enaminone tautomerism. The ratio of tautomers is dependent on the nature of the phenyl ring substituent and the Hammett substituent constants sigma. Theoretical calculations indicate the presence of ketimine and enaminone tautomers in the gas phase. The electron ionization mass spectra of eight 2-phenacylquinolines (ketimine form) were recorded at 70 eV in order to determine the fragmentation routes and to screen for the presence of their enaminone tautomers, (Z)-2-benzoylmethylene-1,2-dihydroquinolines, in the gas phase. The relative abundances or total ion currents of some ions correlated with the Hammett substituent constants and Hammett-Brown constants. The product ions [M-CO](+.) and [M-HCO](+) were observed. A reaction mechanism is suggested for the formation of these ions, requiring skeletal rearrangements. The results furnish information relating to tautomerism in the gas phase.

Environmental pollution affects the plumage color of Great tit nestlings through carotenoid availability.

Birds need to acquire carotenoids for their feather pigmentation from their diet, which means that their plumage color may change as a consequence of human impact on their environment. For example, the carotenoid-based plumage coloration of Great tit, Parus major, nestlings is associated with the degree of environmental pollution. Breast feathers of birds in territories exposed to heavy metals are less yellow than those in unpolluted environments. Here we tested two hypotheses that could explain the observed pattern: (I) deficiency of carotenoids in diet, and (II) pollution-related changes in transfer of carotenoids to feathers. We manipulated dietary carotenoid levels of nestlings and measured the responses in plumage color and tissue concentrations. Our carotenoid supplementation produced the same response in tissue carotenoid concentrations and plumage color in polluted and unpolluted environments. Variation in heavy metal levels did not explain the variation in tissue (yolk, plasma, and feathers) carotenoid concentrations and was not related to plumage coloration. Instead, the variation in plumage yellowness was associated with the availability of carotenoid-rich caterpillars in territories. Our results support the hypothesis that the primary reason for pollution-related variation in plumage color is carotenoid deficiency in the diet.

Substituent effects on the ring-chain tautomerism of some 1,3-oxazolidine derivatives.

The 14 and 70 eV electron ionization mass spectra of five sets (R1 = Me, Et, i-Pr, t-Bu and Ph) of seven 2-aryl-4-R1-substituted (Ar = C6H4X; X = p-NO2, m-Br, p-Cl, H, p-Me, p-OMe and p-NMe2) (1-5) and of seven 2-aryl-5-phenyl-substituted 1,3-oxazolidines (6; for Ar, see above) were recorded to study their ring-chain equilibria in the gas phase. These equilibria were also studied by 1H NMR spectroscopy in CDCl3 for compounds 5 and 6. A few 2,4- and 2,5-dimethyl-2-aryl derivatives (7, 8: Ar = C6H4X; X = m-Br, H and p-OMe) were studied both in CDCl3 and in the gas phase. The main characteristics of the ring-chain equilibria expressed by the variable SigmaRA% of the ring and of the chain form proved to be a strong dependence on the nature of the substituents on C-2 and C-4. The results in the gas phase are compared with those in CDCl3.

3-Oxo-1,3-oxathiolanes-synthesis and stereochemistry.

The compound 3-oxo-1,3-oxathiolane (6) and its cis - and trans - 2-methyl (7,8), 4-methyl (9,10), 5-methyl (11,12) and 2-p-nitrophenyl (13,14) derivatives were prepared by oxidizing the corresponding 1,3-oxathiolanes (1-5) with NaIO4 in water. The oxides were purified and their isomers separated using thin layer chromatography. The structural characterization was carried out with 1H NMR spectroscopy and molecular modeling. Compounds 8-10, 12, and 14 attain the half-chair type conformation with O1 above and C5 below the plane (HC1), but 6, 7, 11, and 13 are, for steric reasons, the mixtures with the alternative half-chair (HC2), where O1 is below and C5 above this plane. Accordingly, 6 appears based on the values of experimental coupling constants as an 81:19, 7 as a 37:63, 11 as a 33:67 and 13 as a 54:46 mixture of HC1 and HC2, respectively. The relative energies of these conformations, the values of the vicinal H,H-coupling constants and 1H chemical shifts were estimated for compounds 6-12 also by computational methods and they support nicely the conclusions based on experimental data.

Substituent effects on 13C chemical shifts of alkyl-substituted 4-oxo-1,3-dioxolanes and 5-oxo-1,3-oxathiolanes.

The 13C NMR chemical shifts for 4-oxo-1,3-dioxolane (1) and its all methyl-substituted derivatives (2-10) as well as for 5-oxo-1,3-oxathiolane (11) and its nine alkyl-substituted derivatives (12-20) are reported. The magnitude and variety of the substituent effects are in accordance with the envelope conformations in which the oxygen or sulfur atom locates at the tip of the envelope as postulated on the basis of earlier data.

Two new phenylbutanoids from inner bark of Betula pendula.

Two phenylbutanoids, 7-{3R-[(4-hydroxyphenyl)butyl] beta-glucopyranosid-O-6-yl} 4-O-beta-glucopyranosylvanillin and 3-beta-glucopyranosyloxy-1-(4-hydroxyphenyl)-butanone were isolated from an aqueous methanol extract of the inner bark of Betula pendula. Their structures were determined by NMR spectroscopy and mass spectrometry. The complete assignment of proton and carbon signals was achieved by 1D and 2D NMR experiments: selective 1D TOCSY, HSQC, HMBC and DQF-COSY.

Electron ionization mass spectra of aryl- and benzyl-substituted 2,3-dihydroimidazo[1,2-a]pyrimidine-5,7(1H,6H)-diones.

The electron ionization mass spectra of the 1-phenyl-, 1-benzyl- and 6-benzyl-1-phenyl-2,3-dihydroimidazo[1,2-a]pyrimidine-5,7(1H,6H)-dione derivatives were recorded at 70 eV to find out the effects of substituents on their fragmentations. Fragmentation pathways were studied using B/E and B(2)/E scans. Some fragmentations involved the loss of C(3)HO(2) or carbon suboxide. The possibility of keto-enol tautomerism was also studied. For comparison selected compounds were studied using (1)H and (13)C NMR spectroscopy to reveal the presence of possible tautomerism. Some ions including [M-OH](+) and [M-HCO](+) and NMR results indicate that the enol form is predominant both in the gas and in the liquid phase.

Substituent effects on the gas-phase ring-chain tautomerism of 3,4,5,6-tetrahydro-2H-1,3-oxazines.

The electron ionization mass spectra of five series of seven 2-aryl,4-R-substituted (R = Me, Et, i-Pr, t-Bu or Ph) 3,4,5,6-tetrahydro-2H-1,3-oxazines were recorded at 14 and 70 eV in order to study the ring-chain tautomeric equilibria in the gas phase. Certain fragment ions were associated with the ring or with the open-chain forms of the compounds. As in chloroform solution, the electron-withdrawing effect of the aryl substituent (p-NO(2), m-Br, p-Cl, H, p-Me, p-OMe and p-NMe(2)) shifts the equilibrium towards the ring form. The correlation of ring-chain equilibria (log K = [ring]/[chain]) with the Hammett sigma+ constants of the aryl substituents was in general good or satisfactory although in some cases the p-NMe(2) did not fit these correlations.

High-performance liquid chromatographic analysis of phlorotannins from the brown alga Fucus vesiculosus.

Separating individual compounds by HPLC represents an effective method for the detection and quantification of phenolic compounds and has been widely utilised. However, phlorotannins are commonly quantified using colorimetric methods, as the total amount of the whole compound group. In the present paper the separation of a set of individual soluble phlorotannins from the phenolic crude extract of Fucus vesiculosus was achieved by HPLC with UV photodiode array detection. Different gradient programs for reversed- and normal-phase HPLC methods were developed and tested. Normal-phase (NP) conditions with a silica stationary phase and a mobile phase with a linear gradient of increasing polarity were found to separate 16 individual components of the phenolic extract. The suitability of the NP-HPLC method for mass spectrometric application was preliminarily tested. Sample preparation was found to be a critical step in the analysis owing to the rapid oxidation of phlorotannins; ascorbic acid was used as an antioxidant.

Reversed-phase HPLC-ESI/MS analysis of birch leaf proanthocyanidins after their acidic degradation in the presence of nucleophiles.

Mountain birch leaves contain large amounts of structurally variable polymeric proanthocyanidins. Their isolation procedure was enhanced by the addition of liquid-liquid extractions prior to column chromatography over Sephadex LH-20. Isolated polymeric proanthocyanidins were depolymerised by acid-catalysis in the presence of benzyl mercaptan or phloroglucinol in order to study their composition. The resulting degradation products, flavan-3-ols and flavan-3-ol adducts, were analysed with reversed-phase high-performance liquid chromatography using UV photodiode array detection for quantification and electrospray ionisation mass spectrometry for identification. The results showed that polymeric proanthocyanidins contained (epi)gallocatechins and (epi)catechins as the extension units and, mainly, (+)-catechin as the terminal unit. The mean degree of polymerisation was found to be 26 based on thiolysis and 31 based on phloroglucinol degradation.

Advanced TMAH and TMAAc thermochemolysis-pyrolysis techniques for molecular characterization of size-separated fractions from aquatic dissolved organic matter.

The structural similarities and differences between the original DOM and the eight size fractions separated were studied in detail with the pyrolysis technique in combination with gas chromatography and mass spectrometry (Py-GC-MS) using two alkylating reagents: TMAH (tetramethylammonium hydroxide), to find both esterified and free carboxylic acids; and TMAAc (tetramethylammonium acetate), to specify only free carboxylic acids. A statistical analysis of the original multidimensional TMAH and TMAAc pyrograms disclosed that the overall structural compositions of the five most important size fractions, accounting for 84% of the original DOM, greatly resembled each other. The remaining three minor size fractions were not classified as homogeneous associations, but they also contained the same total, covalently bound and free carboxylic acid species as the other size fractions and the original DOM mixture, thus representing some kind of intermediate forms. This fundamental outcome strongly supports the opinion that the native dissolved humic-like macromolecules resemble supramolecular associations of smaller molecular size moieties with similar structural functionalities. The concentrations of free aliphatic and aromatic dicarboxylic acids in the DOM solution were so low that their effects on the potential formation of multiply charged ions in electrospray ionization-MS (ESI-MS) studies are likely insignificant.

Does the electron ionization induced fragmentation of partly saturated stereoisomeric pyrrolo- and isoindoloquinazolinones show stereospecificity?

The electron ionization mass spectrometric behavior of pyrroloquinazolinones (1-6) and isoindoloquinazolinones (7-14) was studied. These compounds were further classified as partly saturated pyrroloquinazolinones (1-3), benzologues (7-11), methylene-bridged derivatives (4-6, 12, 13) and a bisacyl compound (14). The mass spectra of the pyrrolo- and isoindoloquinazolinones did not exhibit stereospecific retro-Diels-Alder (RDA) fragmentations. The cyclohexane-fused compounds 7 (cis annelated) and 8 (trans annelated) did display some other ions differing in their abundances that could be used to differentiate this pair of stereoisomers. Also the cyclohexene-fused compounds 2, 3, 9 and 10 exhibited somewhat different ion abundances pairwise that could be utilized for isomeric differentiation. Earlier hypothesis of pyrrolo ring cleavage via the loss of C(3)H(5)O(.) was strengthened by the fragmentation of compounds 1-4. RDA(+/-H) fragmentation is more favorable than the formation of [M-R](+) ions (R=H, C(6)H(4)CH(3), or C(6)H(4)Cl) when an unsaturated bicyclic group is present but both RDA fragmentation and [M-R](+) formation occur for cyclohexene-fused compounds, possibly because of the lower ring strain than with norbornene-fused compounds. The [M-H](+) ion was abundant for compounds 7 and 8 as was [M-Ar](+) for 1-4 and 11. Although the compounds studied might participate in amide-imidol tautomerism, no indication of such tautomerism was detected.

Electron ionization mass spectra of phosphorus-containing heterocycles. III. 1,3,4,2-oxadiazaphosphinane 2-oxides.

The 1,3,4,2-oxadiazaphosphinane 2-oxides differ not only in the relative configuration of the P atom (R* or S*), but also in many other ways such as the ring size, ring fusion, P substitution and bridgehead N atom whose effects on their fragmentations have been studied. Some fragmentations resembled those of 3,1,2-oxazaphosphinane 2-oxides and 1,3,2-diazaphosphinane-2-oxides, but new routes were also found, initiated by ionization at the bridgehead N atom. The relative abundances of the molecular ions and some fragment ions allowed the differentiation of cis-trans epimers. Compounds with n = 2 and R=N(CH(2)CH(2)Cl)(2), and linearly or angularly isoquinoline-fused isomers in most cases, gave more numerous ions with lower relative abundances than the other compounds in this series. Only the isoquinoline derivatives provided fragments resulting from ionization of the aromatic part of the molecule.