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Anhydrolutein I (= (all-E,3R,6'R)-3',4'-didehydro-beta,gamma-caroten-3-ol; 2) and anhydrolutein II (= (all-E, 3R,6'S)-2',3'-didehydro-beta,epsilon-caroten-3-ol; 3) have been isolated and characterized from the extract of steam-cooked sorrel. The presence of these compounds in cooked vegetable is postulated to be due to acid-catalyzed dehydration of lutein (1; Scheme). The structures of the isolated anhydroluteins were established by UV/VIS, CD, and 1H-NMR spectroscopy, and mass spectrometry.

Some cyclic chalcone analogues [E-2-(X-benzylidene)-1-indanones, -tetralones, and -benzosuberones], on-plate UV light-catalyzed formation of new chromatographic spots, can be observed during thin-layer chromatography (TLC). Gas chromatographic (GC) analysis of selected derivatives indicates the formation of one new substance in each case. GC coupled with mass spectrometry and 1 H NMR analysis of the samples reveals that the compounds formed are the respective Z-2-(X-benzylidene)-1-indanones, -tetralones, and -benzosuberones. Two-dimensional TLC shows that the E/Z isomerization is a reversible process. By means of the RP-TLC, the logarithm of n-octanol-water partition coefficient (log P) values of E- and Z-isomeric pairs of selected 2-(X-benzylidene)-1-benzosuberones is determined. The Z-isomers are less lipophilic than the E-isomers.

Starting from 2-nitroadamantane, spiro[pyrrolidine-2,2'-adamantane] nitrones and nitroxides were synthesised as new lipophilic spin traps and spin labels. The reduction of nitrones and nitroxides offers a new route to aminoadamantane derivatives.

Circular dichroism (CD) spectroscopy was used to distinguish between the isomeric (all-E)-configured 3′-epilutein (2) and 6′-epilutein (8) to establish the absolute configuration of epilutein samples of different (natural and semisynthetic) origin, including samples of 2 obtained from thermally processed sorrel. Thus, the CD data of lutein (1) and epilutein samples (2) were compared. Our results unambiguously confirmed the (3R,3′S,6′R)-configuration of all epilutein samples. Compound 2 was thoroughly characterized, and its 13C-NMR data are published herewith for the first time.

Paramagnetic and diamagnetic flavone and flavanone derivatives modified on the C or B rings were synthesized by condensation, Sonogashira reaction, lithiation, and the Baker-Venkataraman procedure.

The isolation, semisynthesis and structure elucidation of carotenoid 5,6-, 5,8- and 3,6-epoxides as well as their derivatives are discussed in detail. This review gives a brief summary of the plausible biosynthetic transformations of the 5,6- epoxy-β-ring. A wide variety of carotenoid end-groups can be obtained from 5,6-epoxy carotenoids. In order to get information on the enzyme activities and regulation of the biosynthetic pathway of carotenoids some of the intermediate steps of these transformations were investigated (e.g. formation of allenic, κ- and γ-end-groups as well as ring opening reactions). The ring opening of the 3-hydroxy-5,6-epoxy end group, resulting in the formation of 3,5,6- trihydroxy compounds, was studied in detail in order to obtain evidence for the proposed reaction mechanisms of the two different biosynthetic routes (acid or enzyme catalysis) which also explain the configurations at the C(5) and C(6) atoms.

The lack of efficacy of anti-inflammatory drugs, anti-coagulants, anti-oxidants, etc. in critically ill patients has shifted interest towards developing alternative treatments. Since inhibitors of the nuclear enzyme poly-(ADP-ribose) polymerase (PARP) were found to be beneficial in many pathophysiological conditions associated with oxidative stress and PARP-1 knock-out mice proved to be resistant to bacterial lipopolysaccharide (LPS)-induced septic shock, PARP inhibitors are candidates for such a role. In this study, the mechanism of the protective effect of a potent PARP-1 inhibitor, PJ34 was studied in LPS-induced (20 mg/kg, i.p.) septic shock in mice. We demonstrated a significant inflammatory response by magnetic resonance imaging in the dorsal subcutaneous region, in the abdominal regions around the kidneys and in the inter-intestinal cavities. We have found necrotic and apoptotic histological changes as well as obstructed blood vessels in the liver and small intestine. Additionally, we have detected elevated tumor necrosis factor-α levels in the serum and nuclear factor kappa B activation in liver of LPS-treated mice. Pre-treating the animals with PJ34 (10 mg/kg, i.p.), before the LPS challenge, besides rescuing the animals from LPS-induced death, attenuated all these changes presumably by activating the phosphatidylinositol 3-kinase–Akt/protein kinase B cytoprotective pathway.

A series of new 4-(arylmethylene)-3-isochromanones 1−17 has been prepared by base-catalyzed Knoevenagel condensations. Stereochemical and conformational analyses were performed by 1H and 13C NMR methods, and X-ray crystallography was used to complement the configurational assignment for a representative product. The reaction generally yields the (E) isomer, but the isomeric composition of the products was influenced by the aromatic aldehyde and usually governed by the stability of the products. Several aldehydes exclusively afforded the (Z) isomer, due to intramolecular steric/electronic interactions reflected in the increased stability of the product. The condensation performed with ortho-hydroxyarenecarbaldehydes furnished the unexpected 3-arylcoumarins or 3-arylbenzocoumarins. (© Wiley-VCH Verlag GmbH, 69451 Weinheim, Germany, 2002)

Glucopyranosylidene spirothiohydantoin (TH) has been identified as a potential inhibitor of both muscle and liver glycogen phosphorylase b (GPb) and a (GPa) and shown to diminish liver GPa activity in vitro. Kinetic experiments reported here show that TH inhibits muscle GPb competitively with respect to both substrates phosphate (K(i)=2.3 microM) and glycogen (K(i)=2.8 microM). The structure of the GPb-TH complex has been determined at a resolution of 2.26 A and refined to a crystallographic R value of 0.193 (R(free)=0.211). The structure of GPb-TH complex reveals that the inhibitor can be accommodated in the catalytic site of T-state GPb with very little change of the tertiary structure, and provides a basis of understanding potency and specificity of the inhibitor. The glucopyranose moiety makes the standard hydrogen bonds and van der Waals contacts as observed in the glucose complex, while the rigid thiohydantoin group is in a favourable electrostatic environment and makes additional polar contacts to the protein.

The aim of our study was to establish a simple in vivo method for water quantification in vasogenic edema, and provide data on imaging of mouse brain at 9.4 Tesla. Apparent T1 and spin density values determined by MRI were found to strongly correlate with the gravimetric water content of mouse brain undergoing cold injury. Using a two-point calibration line between the spin density values for pure water and cortex of mouse brain, as well as the corresponding water contents in vivo, water could be quantified with satisfactory accuracy. Magn Reson Med 46:1246–1249, 2001. © 2001 Wiley-Liss, Inc.

D-Gluco- and D-xylopyranosylidene-spiro-hydantoins and -thiohydantoins were prepared from the parent sugars in a six-step, highly chemo-, regio-, and stereoselective procedure. In the key step of the syntheses C-(1-bromo-1-deoxy-beta-D-glycopyranosyl)formamides were reacted with cyanate ion to give spiro-hydantoins with a retained configuration at the anomeric center as the major products. On the other hand, thiocyanate ions gave spiro-thiohydantoins with an inverted anomeric carbon as the only products. On the basis of radical inhibition studies, a mechanistic rationale was proposed to explain this unique stereoselectivity and the formation of C-(1-hydroxy-beta-D-glycopyranosyl)formamides as byproducts. Enzyme assays with a and b forms of muscle and liver glycogen phosphorylases showed spiro-hydantoin 12 and spiro-thiohydantoin 14 to be the best and equipotent inhibitors with K(i) values in the low micromolar range. The study of epimeric pairs of D-gluco and D-xylo configurated spiro-hydantoins and N-(D-glucopyranosyl)amides corroborated the role of specific hydrogen bridges in binding the inhibitors to the enzyme.

Ischemia-reperfusion induces reactive oxygen species (ROS) formation, and ROS lead to cardiac dysfunction, in part, via the activation of the nuclear poly(ADP-ribose) polymerase (PARP, called also PARS and ADP-RT). ROS and peroxynitrite induce single-strand DNA break formation and PARP activation, resulting in NAD(+) and ATP depletion, which can lead to cell death. Although protection of cardiac muscle by PARP inhibitors can be explained by their attenuating effect on NAD(+) and ATP depletion, there are data indicating that PARP inhibitors also protect mitochondria from oxidant-induced injury. Studying cardiac energy metabolism in Langendorff heart perfusion system by (31)P NMR, we found that PARP inhibitors (3-aminobenzamide, nicotinamide, BGP-15, and 4-hydroxyquinazoline) improved the recovery of high-energy phosphates (ATP, creatine phosphate) and accelerated the reutilization of inorganic phosphate formed during the ischemic period, showing that PARP inhibitors facilitate the faster and more complete recovery of the energy production. Furthermore, PARP inhibitors significantly decrease the ischemia-reperfusion-induced increase of lipid peroxidation, protein oxidation, single-strand DNA breaks, and the inactivation of respiratory complexes, which indicate a decreased mitochondrial ROS production in the reperfusion period. Surprisingly, PARP inhibitors, but not the chemically similar 3-aminobenzoic acid, prevented the H(2)O(2)-induced inactivation of cytochrome oxidase in isolated heart mitochondria, suggesting the presence of an additional mitochondrial target for PARP inhibitors. Therefore, PARP inhibitors, in addition to their important primary effect of decreasing the activity of nuclear PARP and decreasing NAD(+) and ATP consumption, reduce ischemia-reperfusion-induced endogenous ROS production and protect the respiratory complexes from ROS induced inactivation, providing an additional mechanism by which they can protect heart from oxidative damages.

In an investigation of carotenoids present in the fruits ofAsparagus falcatus capsanthin (1), capsorubin (2), 5,6-diepikarpoxanthin (7), capsanthin 5,6-epoxide (18), capsochrome (17), mutatoxanthin (19), antheraxanthin (11), and capsanthone (20) (Figure 1) have been isolated by preparative CLC and characterized by spectroscopic methods. On the basis of spectroscopic data the absolute configuration of 5,6-diepikarpoxanthin (7) was determined as 3S,5S,6S, which is identical with that occurring in samples originating from paprika andLilium.

One-, two- and three-bond carbon-13; proton scalar spin-spin coupling constants (1JC,H, 2JC,H,3JC,H) have been determined for a series of hydantoin- and 2-thiohydantoin derivatives in DMSO-d6 solution using one- and two-dimensional NMR techniques. Simultaneous consideration of several coupling data allowed reliable estimates to be deduced for the ring dihedrals equivalent to the ϕ, ψ and ω angles in peptides. These data are in support of a quasi-planar geometry for the hydantoin ring in solution in close agreement with previous results obtained for the solid state. Rotamer population distributions around the C5C1′ bond have been determined for some 5-substituted derivatives. The existence of four-bond couplings between amide NH protons in 1,3-positions of the imidazolidine ring has been demonstrated for the first time.

Partial hydrolysis of the nitrile moiety in acetylated 1-bromo-glycopyranosyl cyanides 1 and 9 mediated by TiCl4 gave C-(1-bromo-1-deoxy-glycopyranosyl)formamides 2 and 10 whose reaction either with AgOCN, AgSCN or KSCN in nitromethane resulted in the formation of glycopyranosylidene-spirohydantoins 3, 11 and 15 and -thiohydantoins 7 and 16. Zemplen-deacetylation gave the pyranoid epi-hydantocidin analogues 4 and 12, and thiohydantocidin analogues 8 and 17, respectively.

Partial hydrolysis of the nitrile moiety in acetylated 1-bromo-glycopyranosyl cyanides 1 and 9 mediated by TiCl4 gave C-(1-bromo-1-deoxy-glycopyranosyl)formamides 2 and 10 whose reaction either with AgOCN, AgSCN or KSCN in nitromethane resulted in the formation of glycopyranosylidene-spirohydantoins 3, 11 and 15 and -thiohydantoins 7 and 16. Zemplen-deacetylation gave the pyranoid epi-hydantocidin analogues 4 and 12, and thiohydantocidin analogues 8 and 17, respectively.

In the present work, we show that in processed sorrel, the lutein (1) can convert to 3’-epilutein (2) – 3’-stereoisomer of lutein – by epimerisation reaction, as well as to anhydrolutein I (3) and II (4) by dehydration reaction. The newly formed anhydrolutein I (3) and II (4) were detected and identified by HPLC-DAD and HPLC-MS techniques and co-chromatography with authentic samples. The compounds 1 and 2 were also isolated from cooked sorrel by column chromatography and characterized by NMR spectroscopy.

Activation of the nuclear enzyme poly(ADP-ribose) polymerase (PARP) is involved in numerous pathophysiological conditions. Because PARP-1 knockout mice are resistant to endotoxin-induced shock and inhibitors of the enzyme were reported to have similar beneficial properties, we investigated the effect of 4-hydroxyquinazoline (4-HQN), a potent PARP-1 inhibitor, on the modulation of kinase cascades and the regulation of transcription factors in a rodent septic shock model. T2-weighted magnetic resonance imaging showed the pattern of anatomical localization of the inflammatory response in bacterial lipopolysaccharide (LPS)-treated mice and the anti-inflammatory effect of the PARP-1 inhibitor. We have found that 4-HQN activated the phosphatidylinositol 3 (PI3)-kinase/Akt pathway in lung, liver, and spleen, and down-regulated two elements of the MAP kinase system. Namely, it dramatically attenuated the activation of the LPS-induced extracellular signal-regulated kinase (ERK)1/2 and p38 mitogen-activated protein (MAP) kinase in a tissue-specific manner. Furthermore, phosphorylation of p90RSK, a downstream target of ERK1/2, showed a similar pattern of down-regulation as did the phosphorylation of ERK1/2 and p38 after LPS and 4-HQN treatment. As a consequence of the aforementioned effects on the kinase pathways, 4-HQN decreased the activation of transcription factor nuclear factor-kappaB (NF-kappaB) and activator protein 1 (AP-1) in LPS-induced endotoxic shock. Our results provide evidence for the first time that the beneficial effects of PARP inhibition in endotoxic shock, such as attenuation of NF-kappaB- and AP-1 transcription factor activation, are mediated, at least partially, through the regulation of the PI3-kinase/Akt pathway and MAP kinase cascades.

To prepare effective PARP [poly(ADP)-ribose)polymerase-1] inhibitors, starting from 2-mercapto-4(3H)-quinazolone (1), several S-alkyl derivatives—2-alkylsulfanyl-3H-quinazolin-4-ones (2–5, 7–9)—as well as an S-benzyl derivative (10) were prepared using a simple alkylation method. Some of them are known compounds. Their structure was studied thoroughly by MS and NMR methods.

Lutein can convert to 3′-epilutein and anhydrolutein I in acidic medium as well as during different cooking methods. To our knowledge, this is the first demonstration of the epimerisation and dehydration of lutein in native samples. During different cooking processes the lutein content of the processed vegetables and fruits dramatically decreases, thus the bioavailability of processed plant food may also decrease. The occurrence of 3′-epilutein in processed food may be important regarding the application of synthetic lutein as food additive.

In vivo water content determination based on magnetic resonance (MR) method is of importance in clinical practice as well as in animal studies to follow up the treatment given in order to reduce brain oedema. The methods proposed in the literature so far are largely time consuming. The aim of this study was to find a fast in vivo water quantification method having real advantage for patients suffering from critical conditions.Cold injury was applied to provoke brain oedema in fourteen rats. T(1) values of both the oedematous area and the contralateral normal cortex were determined by two independent methods 24 hours after the cold impact. First, from a series of images recorded by inversion recovery spin echo (IRSE) sequence and then by progressive saturation experiment performed by localised MR spectroscopy using stimulated echo acquisition mode (STEAM). To reduce the acquisition time, a two-element repetition time array was optimised for the STEAM experiment, whereas four inversion times were used for T(1) mapping. Both methods were validated against gel phantoms with known T(1) values. After the MR measurements the animals were sacrificed and the water contents of the regions of interest were determined by gravimetric wet-dry method.The reciprocals of the in vivo measured T(1) values were correlated with the reciprocals of the brain water contents. STEAM experiment showed stronger correlation (r=0.96) than IRSE (r=0.93). In addition, STEAM provided more accurate T(1) values in the phantom study. Determination of brain water content based on T1 measurement does work also at high magnetic field. Determination of brain water content by Magnetic Resonance Spectroscopy is feasible within 2 minutes.Using the presented fast method, water content can be determined within a couple of minutes in animal experiments as well as in the daily clinical practice.

Nephrotoxicity is one of the major dose limiting side effects of cisplatin chemotherapy. The antitumor and toxic effects are mediated in part by different mechanisms, thus, permitting a selective inhibition of certain side effects. The influence of O-(3-piperidino-2-hydroxy-1-propyl)nicotinic amidoxime (BGP-15) - a poly(ADP-ribose) polymerase (PARP) inhibitor - on the nephrotoxicity and antitumor efficacy of cisplatin has been evaluated in experimental models. BGP-15 either blocked or significantly reduced (60-90% in 100-200 mg/kg oral dose) cisplatin induced increase in serum urea and creatinine level in mice and rats and prevented the structural degeneration of the kidney, as well. The nephroprotective effect of BGP-15 treatment was revealed also in living mice by MRI analysis manifesting in the lack of oedema which otherwise developed as a result of cisplatin treatment. The protective effect was accompanied by inhibition of cisplatin-induced poly-ADP-ribosylation and by the restoration of the disturbed energy metabolism. The preservation of ATP level in the kidney was demonstrated in vivo by localized NMR spectroscopy. BGP-15 decreased cisplatin-induced ROS production in rat kidney mitochondria and improved the antioxidant status of the kidney in mice with cisplatin-induced nephropathy. In rat kidney, cisplatin caused a decrease in the level of Bcl-x, a mitochondrial protective protein, and this was normalized by BGP-15 treatment. On the other hand, BGP-15 did not inhibit the antitumor efficacy of cisplatin in cell culture and in transplantable solid tumors of mice. Treatment with BGP-15 increased the mean survival time of cisplatin-treated P-388 leukemia bearing mice from 13 to 19 days. PARP inhibitors have been demonstrated to diminish the consequences of free radical-induced damage, and this is related to the chemoprotective effect of BGP-15, a novel PARP inhibitor. Based on these results, we propose that BGP-15 represents a novel, non-thiol chemoprotective agent.

Reaction of glycosyl trimethylphosphinimides with carbon disulfide under mild conditions (room temperature, short reaction time) leads to symmetrical glycosyl carbodiimides. Addition of bis(trimethylsilyl)carbodiimide to peracetylated aldoses under the influence of SnCl4 afforded N,N-bis(glycosyl)cyanamides for the first time. Readily accessible unsymmetrical N,N′-bis(glycosyl)thioureas can be desulfurated and transformed into the corresponding carbodiimides using HgO in CHCl3/water at room temperature.