Cinnabarinic acid, an endogenous metabolite of the kynurenine pathway, activates type 4 metabotropic glutamate receptors.

Cinnabarinic acid is an endogenous metabolite of the kynurenine pathway that meets the structural requirements to interact with glutamate receptors. We found that cinnabarinic acid acts as a partial agonist of type 4 metabotropic glutamate (mGlu4) receptors, with no activity at other mGlu receptor subtypes. We also tested the activity of cinnabarinic acid on native mGlu4 receptors by examining 1) the inhibition of cAMP formation in cultured cerebellar granule cells; 2) protection against excitotoxic neuronal death in mixed cultures of cortical cells; and 3) protection against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxicity in mice after local infusion into the external globus pallidus. In all these models, cinnabarinic acid behaved similarly to conventional mGlu4 receptor agonists, and, at least in cultured neurons, the action of low concentrations of cinnabarinic acid was largely attenuated by genetic deletion of mGlu4 receptors. However, high concentrations of cinnabarinic acid were still active in the absence of mGlu4 receptors, suggesting that the compound may have off-target effects. Mutagenesis and molecular modeling experiments showed that cinnabarinic acid acts as an orthosteric agonist interacting with residues of the glutamate binding pocket of mGlu4. Accordingly, cinnabarinic acid did not activate truncated mGlu4 receptors lacking the N-terminal Venus-flytrap domain, as opposed to the mGlu4 receptor enhancer, N-phenyl-7-(hydroxyimino)cyclopropa[b]chromen-1a-carboxamide (PHCCC). Finally, we could detect endogenous cinnabarinic acid in brain tissue and peripheral organs by high-performance liquid chromatography-tandem mass spectrometry analysis. Levels increased substantially during inflammation induced by lipopolysaccharide. We conclude that cinnabarinic acid is a novel endogenous orthosteric agonist of mGlu4 receptors endowed with neuroprotective activity.

The influence of collection zone on glucosinolates, polyphenols and flavonoids contents and biological profiles of Capparis sicula ssp. sicula.

This study aimed to evaluate the influence of collection zone on total phenol, flavonoid and glucosinolate contents and antioxidant and anti-inflammatory activities of caper (Capparis sicula ssp. sicula). This species has been characterized through the detection, isolation and quantitative evaluation of chemical markers (polyphenols, flavonoids and glucosinolates). The chemical investigation showed a different composition between the two collection zones. While the total amounts of phenolics and flavonoids of the two samples were quite the same, their high-performance liquid chromatography profiles were very different. In both samples, the most abundant aglycone was quercetin which accounted for 60% of total flavonoids. Nuclear magnetic resonance data analysis allowed the identification of two compounds: 3,5-dicaffeoylquinic and 4,5-dicaffeoylquinic acids which represented 6.67% and 15.94%, respectively, of the total amount of flavonoids in sample 1. In sample 2, these two acids were still present, but their percentages were much less (2.20% and 1.71%, respectively). As far as we know, this is the first report about the presence of dicaffeoylquinic acids in Capparis. With regard to glucosinolate content, sample 1 showed a higher content of glucosinolates. In both samples, glucocapparin was the most abundant compound. Antioxidant activity of the methanolic C. sicula extracts using diphenyl picrylhydrazyl, ß-carotene bleaching test and oxygen radical absorbance capacity showed that the sample 2 was more active than 1. As regards the inhibition of NO production, the extracts from sample 2 were more active than those from sample 1.

In vitro effects of natural prenyloxycinnamic acids on human cytochrome P450 isozyme activity and expression.

Previous studies demonstrated that natural prenyloxyphenylpropanoid derivatives have potent biological properties like anti-cancer effects in vitro and in vivo. Additionally they are extremely safe and associated with low toxicity, making them excellent candidates as chemopreventive agents. However, so far only little is known about possible interactions with isoforms of cytochrome P450 (CYPs) being involved in the metabolism of xenobiotics and representing a major site for drug-drug interactions. The aim of this study was to evaluate the effects of selected natural prenyloxyphenylpropanoids (prenyloxycinnamic acids) on expression and activity of some major CYPs and on the activity of the major drug efflux transporter P-glycoprotein (P-gp). Inhibition of CYP3A4, CYP2C19, and CYP2D6 was quantified using commercially available kits. P-gp inhibtion was quantified by calcein assay. Induction of CYP mRNA (CYP3A4, CYP2C19, CYP2C9, and CYP2B6) was measured in LS180 cells by quantitative real-time reverse transcriptase polymerase chain reaction using the LightCycler technology. Only boropinic acid revealed substantial inhibition of CYPs, especially of CYP2C19 (IC50 = 31±5µM). This compound also had the most pronounced effect on CYP mRNA expression among the prenyloxycinnamic acids tested. However all but 4'-isopentenyloxy-p-coumaric acid revealed inducing effects on CYPs with different induction profiles. P-gp was only significantly inhibited by 4'-geranyloxyferulic acid. This was the first study demonstrating modulating effects of prenyloxycinnamic acids on CYP activity and expression and on P-gp activity. The results suggest that boropinic acid is most prone to drug-drug interactions at the level of CYPs, whereas 4'-isopentenyloxy-p-coumaric acid does not modulate CYP activity and expression.

Quantification of 4'-geranyloxyferulic acid, a new natural colon cancer chemopreventive agent, by HPLC-DAD in grapefruit skin extract.

Oxyprenylated natural products (isopentenyloxy-, geranyloxy- and the less spread farnesyloxy-compounds and their biosynthetic derivatives) represent a family of secondary metabolites that have been consider for years merely as biosynthetic intermediates of the most abundant C-prenylated derivatives. Many of the isolated oxyprenylated natural products were shown to exert in vitro and in vivo remarkable anti-cancer and anti-inflammatory effects. 4'-Geranyloxyferulic acid [3-(4'-geranyloxy-3'-methoxyphenyl)-2-trans-propenoic] has been discovered as a valuable chemopreventive agent of several types of cancer. After development of a high yield and "eco-friendly" synthetic scheme of this secondary metabolite, starting from cheap and non-toxic reagents and substrates, we developed a new HPLC-DAD method for its quantification in grapefruit skin extract. A preliminary study on C18 column showed the separation between GOFA and boropinic acid (having the same core but with an isopentenyloxy side chain), used as internal standard. The tested column were thermostated at 28+/-1 degrees C and the separation was achieved in gradient condition at a flow rate of 1 mL/min with a starting mobile phase of H(2)O:methanol (40:60, v/v, 1% formic acid). The limit of detection (LOD, S/N=3) was 0.5 microg/mL and the limit of quantification (LOQ, S/N=10) was 1 microg/mL. Matrix-matched standard curves showed linearity up to 75 microg/mL. In the analytical range the precision (RSD%) values were

HPLC separation and NMR structural elucidation of sn-1,2-, 2,3-, and 1,3-diacylglycerols from olive oil as naphthylethylurethane derivatives.

In this study, sn-1,2-, sn-2,3-, and sn-1,3-diacylglycerols were isolated from olive oil, and their urethane derivatives (urethanes) were prepared. Normal-phase high-performance liquid chromatography (NP-HPLC) separation of the urethane isomers was performed and the separate classes were studied by nuclear magnetic resonance (NMR). The use of 1H NMR and homo- and heteronuclear 2D techniques provided a great amount of information in a very short time, particularly when a high-field NMR instrument (700 MHz) was used. Particularly diagnostic for this kind of compound was the glyceridic moiety that presents typical chemical shifts both for carbon and hydrogen. These studies show the usefulness of NMR spectroscopy to recognize clearly the sn-1,3- and, moreover, sn-1,2- with respect to sn-2,3-diacylglycerols, although very minor differences occur between them.

Antifungal activity of some Cuban Zanthoxylum species.

Ethanolic extracts of the trunk bark of Zanthoxylum fagara, Z. elephantiasis and Z. martinicense showed activity against different species of fungi. No antibacterial activity was detected.

Evaluation of 137Cs activity in plant drugs and in some phytoderivatives from Chernobyl accident up to present (1986-1994).

The accident to the nuclear power plant of Chernobyl (in April 1986) caused a radioactive contamination in large areas of the majority of European countries. During the first transient time the fall-out phenomenon was the most important method of contamination, particularly from 131I whose relative isotopic abundance with respect to other released radionuclides was very high. Thereafter, 137Cs, owing to its long half-time and its large presence in environmental matrixes and so in the food chain, became the element on which the attention was to be focused. Plant drugs and their derivatives are, at present, of very large alimentary consumption among people. This can cause some problems to human health, so the authors have studied (from 1986 to 1994) the activity of 137Cs in a large number of drugs (about 5000) and in some industrial and home-made officinal products. Some suggestions on the Cs+,K+ ions competition in soil can also be derived.

Inhibition of pig kidney dopa decarboxylase by coenzyme-5-hydroxytryptophan adducts.

The effect of N-(5'-phosphopyridoxyl)-L-5-hydroxytryptophan, N-(5'-phosphopyridoxyl)-D-5-hydroxytryptophan and N-(5'-phosphopyridoxyl)-5-hydroxytryptamine on the reactivation of apoDopa decarboxylase to holoenzyme has been investigated. The different degree of inhibition exerted by these adducts has been interpreted on the basis of a different orientation of the 2 isomers of 5-HTP at the active of Dopa decarboxylase.

Determination of organophosphorus pesticides by thin-layer chromatography.

Analysis for 13 common organophosphorus pesticides by thin-layer chromatography is described; 17 solvent systems were examined. With channel thin-layer chromatography, linear calibration graphs were obtained for the range 1-10 mug.

The preparation of 5-spiro-7'-(bicyclo (4.1.0.) heptane)barbituric acid.

The 7,7-diethoxycarbonylnorcarane (III), obtained by a catalytic decomposition of diethyldiazomalonate (I a) in cyclohexene, was condensed with urea to give the 5-spiro-7'-(bicyclo(4.1.0.)heptane)barbituric acid (IV).

[Derivatives of isothiochromane]. / Derivati dell'isotiocromano

In connection with a program, the purpose of which is to aim at the synthesis of molecules containing the residual phenylethylamine in a rigid structure, the synthesis of 1-methylaminoisotiocromane (IV) starting from isotiocromane (I) is hereby described.