Paula Aulaskari

Synthesis and anti-inflammatory effects of a series of novel 7-hydroxycoumarin derivatives.

A number of 7-hydroxycoumarins have been synthesised by Pechmann cyclisation using differently substituted resorcinols employing perchloric acid as the condensing agent. All the compounds have been characterised by analytical and spectroscopic methods. The anti-inflammatory properties were tested with LPS-induced inflammation in J774 macrophages. Expression of iNOS and COX-2 was determined by Western blot, NO by nitrite assay and IL-6 by ELISA analyses. Fifteen of the tested 7-hydroxycoumarins also inhibited IL-6 production but none of them had any major inhibitory effect on COX-2 expression.

Anti-inflammatory properties of a dual PPARgamma/alpha agonist muraglitazar in in vitro and in vivo models

IntroductionPeroxisome proliferator-activated receptor (PPAR) agonists are widely used drugs in the treatment of diabetes and dyslipidemia. In addition to their metabolic effects, PPAR isoforms PPARα and PPARγ are also involved in the regulation of immune responses and inflammation. In the present study, we investigated the effects of a dual PPARγ/α agonist muraglitazar on inflammatory gene expression in activated macrophages and on carrageenan-induced inflammation in the mouse.MethodsJ774 murine macrophages were activated by lipopolysaccharide (LPS) and treated with dual PPARγ/α agonist muraglitazar, PPARγ agonist GW1929 or PPARα agonist fenofibrate. The effects of PPAR agonists on cytokine production and the activation of inducible nitric oxide synthase (iNOS) pathway were investigated by ELISA, Griess method, Western blotting and quantitative RT-PCR. Nuclear translocation, DNA-binding activity and reporter gene assays were used to assess the activity of nuclear factor kappa B (NF-kB) transcription factor. Carrageenan-induced paw oedema was used as an in vivo model of acute inflammation.ResultsMuraglitazar as well as PPARγ agonist GW1929 and PPARα agonist fenofibrate inhibited LPS-induced iNOS expression and NO production in activated macrophages in a dose-dependent manner. Inhibition of iNOS expression by muraglitazar included both transcriptional and post-transcriptional components; the former being shared by GW1929 and the latter by fenofibrate. All tested PPAR agonists also inhibited IL-6 production, while TNFα production was reduced by muraglitazar and GW1929, but not by fenofibrate. Interestingly, the anti-inflammatory properties of muraglitazar were also translated in vivo. This was evidenced by the finding that muraglitazar inhibited carrageenan-induced paw inflammation in a dose-dependent manner in mice as did iNOS inhibitor L-NIL and anti-inflammatory steroid dexamethasone.ConclusionsThese results show that muraglitazar has anti-inflammatory properties both in vitro and in vivo and these effects reflect the agonistic action through both PPARα and PPARγ.

Low-energy collision-induced dissociation tandem mass spectrometry of 7-acetonyloxycoumarins

RATIONALE Coumarins are naturally occurring, oxygen-containing heterocycles with considerable pharmaceutical potential. For structural elucidation of natural or synthetic coumarins, tandem mass spectrometry (MS(n)) represents an essential tool. In this study, fragmentation characteristics of twenty-two 7-acetonyloxycoumarins, having promising anti-inflammatory properties, were investigated with low-energy collision-induced dissociation (CID). METHODS Accurate mass measurements were performed on a 12-T Fourier transform ion cyclotron resonance (FT-ICR) instrument. Most CID-MS(n) measurements were performed on a quadrupole ion trap (QIT) instrument, except some additional CID-MS(2) measurements performed on the FT-ICR instrument for further confirmation of some fragment ions. Positive-ion electrospray ionization (ESI) was employed throughout. Density functional theory (DFT) calculations (B3LYP) were carried out to analyze putative ion structures/fragmentation channels. RESULTS The most favourable dissociation channel for [M + H](+) ions of 7-acetonyloxycoumarins was the elimination of a C3H5O(●) radical (57 Da) from the 7-acetonyloxy group via homolytic bond cleavage. The resulting phenolic radical ion was the primary fragment ion for the most compounds studied. Losses of even-electron neutrals, C3H4O and C3H6O (56 and 58 Da), were also observed. These primary eliminations were accompanied with other characteristic neutral losses from the coumarin skeleton, including H2O, CO, CO2, and C2H2O (ketene). In addition, propene (C3H6) loss was also observed for 4-propyl or 3-ethyl-4-methyl-substituted compounds. CONCLUSIONS The studied coumarins showed interesting characteristics in low-energy CID due to the presence of a 7-acetonyloxy group, leading to both even- and odd-electron product ions. The main dissociation channels observed for each compound were highly dependent on the substituents in the benzopyranone ring. The present results will advance our knowledge on the dissociation characteristics of both synthetic and natural coumarins.

Negative ion electrospray ionization mass spectrometry and computational studies on substituted 7-hydroxycoumarins.

Twenty-two substituted 7-hydroxycoumarins were studied by negative ion electrospray ionization collision-induced dissociation (CID) mass spectrometry. Fragmentation pathways were also investigated by computation method using the B3LYP density functional theory. In general, the most important fragmentations of the 7-hydroxycoumarin [M – H]− ions were the elimination of CO2 and CO which agreed with the calculated energies of the proposed fragmentation reactions. In most cases, methyl group elimination was also favorable. Methyl group elimination occurred in three different ways, the most interesting being hydrogen rearrangement from a neighboring alkyl group to a ring carbon, which led to a benzyl radical formation. In some cases, CH2CO elimination was observed as well. Isomeric compounds gave rise to different CID spectra.

Improved synthesis and characterization of 1,3,4,6-tetra-O-acetyl-2-(N-acetylacetamido)-2-deoxy-β-d-glucopyranose

A method from the 1960s to synthesize the N,N-diacetyl derivative of peracetylated beta-D-glucosamine was improved by assistance of molecular sieves. The melting point of the title compound was revised and the structure determined by means of X-ray diffraction.

Synthesis of 1-Substituted-5-[(2-nitro-2-phenyl)-ethyl]imidazoles

Abstract 1-R-5-[(2-Nitro-2-phenyl)ethyl]imidazoles 9a,b,c (R = Bn, Me, H) have been prepared from the corresponding imidazole nitroethylenes 6a,b,c by the reduction with sodium borohydride in THF-methanol.