A Study about Regioisomeric Hydroquinones with Multiple Intramolecular Hydrogen Bonding.

A theoretical exploration about hydrogen bonding in a series of synthetic regioisomeric antitumor tricyclic hydroquinones is presented. The stabilization energy for the intramolecular hydrogen bond (IHB) formation in four structurally different situations were evaluated: (a) IHB between the proton of a phenolic hydroxyl group and an ortho-carbonyl group (forming a six-membered ring); (b) between the oxygen atom of a phenolic hydroxyl group and the proton of an hydroxyalkyl group (seven membered ring); (c) between the proton of a phenolic hydroxyl group with the oxygen atom of the hydroxyl group of a hydroxyalkyl moiety (seven-membered ring); and (d) between the proton of a phenolic hydroxyl group and an oxygen atom directly bonded to the aromatic ring in ortho position (five-membered ring). A conformational analysis for the rotation around the hydroxyalkyl substituent is also performed. It is observed that there is a correspondence between the conformational energies and the IHB. The strongest intramolecular hydrogen bonds are those involving a phenolic proton and a carbonyl oxygen atom, forming a six-membered ring, and the weakest are those involving a phenolic proton with the oxygen atom of the chromenone, forming five-membered rings. Additionally, the synthesis and structural assignment of two pairs of regioisomeric hydroquinones, by 2D-NMR experiments, are reported. These results can be useful in the design of biologically-active molecules.

Inversion of the stereochemical configuration (3S, 5S)-clavaminic acid into (3R, 5R)-clavulanic acid: A computationally-assisted approach based on experimental evidence.

Clavulanic acid (CA), a potent inhibitor of ß-lactamase enzymes, is produced by Streptomyces clavuligerus (Sc) cultivation processes, for which low yields are commonly obtained. Improved knowledge of the clavam biosynthetic pathway, especially the steps involved in the inversion of 3S-5S into 3R-5R stereochemical configuration, would help to eventually identify bottlenecks in the pathway. In this work, we studied the role of acetate in CA biosynthesis by a combined continuous culture and computational simulation approach. From this we derived a new model for the synthesis of N-acetyl-glycyl-clavaminic acid (NAG-clavam) by Sc. Acetylated compounds, such as NAG-clavam and N-acetyl-clavaminic acid, have been reported in the clavam pathway. Although the acetyl group is present in the ß-lactam intermediate NAG-clavam, it is unknown how this group is incorporated. Hence, under the consideration of the experimentally proven accumulation of acetate during CA biosynthesis, and the fact that an acetyl group is present in the NAG-clavam structure, a computational evaluation of the tentative formation of NAG-clavam was performed for the purpose of providing further understanding. The proposed reaction mechanism consists of two steps: first, acetate reacts with ATP to produce a reactive acylphosphate intermediate; second, a direct nucleophilic attack of the terminal amino group of N-glycyl-clavaminic on the carbonyl carbon of the acylphosphate intermediate leads to a tetrahydral intermediate, which collapses and produces ADP and N-acetyl-glycyl-clavaminic acid. The calculations suggest that for the proposed reaction mechanism, the reaction proceeds until completion of the first step, without the direct action of an enzyme, where acetate and ATP are involved. For this step, the computed activation energy was ≅2.82kcal/mol while the reaction energy was ≅2.38kcal/mol. As this is an endothermic chemical process with a relatively small activation energy, the reaction rate should be considerably high. The calculations offered in this work should not be considered as a definite characterization of the potential energy surface for the reaction between acetate and ATP, but rather as a first approximation that provides valuable insight about the reaction mechanism. Finally, a complete route for the inversion of the stereochemical configuration from (3S, 5S)-clavaminic acid into (3R, 5R)-clavulanic acid is proposed, including a novel alternative for the double epimerization using proline racemase and NAG-clavam formation.

Synthesis, leishmanicidal and cytotoxic activity of triclosan-chalcone, triclosan-chromone and triclosan-coumarin hybrids.

Twelve hybrids derived from triclosan were obtained via Williamson etherification of O-triclosan alkyl bromide plus chalcone and O-coumarin or O-chromone alkyl bromide plus triclosan, respectively. Structures of the products were elucidated by spectroscopic analysis. The synthesized compounds were evaluated for antileishmanial activity against L. (V) panamensis amastigotes. Cytotoxic activity was also evaluated against mammalian U-937 cells. Compounds 7-9 and 17, were active against Leishmania parasites (EC50=9.4; 10.2; 13.5 and 27.5 µg/mL, respectively) and showed no toxicity toward mammalian cells (>200 µg/mL). They are potential candidates for antileishmanial drug development. Compounds 25-27, were active and cytotoxic. Further studies using other cell types are needed in order to discriminate whether the toxicity shown by these compounds is against tumor or non-tumor cells. The results indicate that compounds containing small alkyl chains show better selectivity indices. Moreover, Michael acceptor moieties may modify both the leishmanicidal activity and cytotoxicity. Further studies are required to evaluate if the in vitro activity against Leishmania panamensis demonstrated here is also observed in vivo.

NMR assignment in regioisomeric hydroquinones.

A set of regioisomeric pairs of tricyclic hydroquinones, analogues of antitumor 9,10-dihydroxy-4,4-dimethyl-5,8-dihydroanthracen-1(4H)-one (1) and other derivatives, were synthesized and their regiochemistry and NMR spectra assigned by using (1)H-detected one-bond (C-H) HMQC and long-range C-H HMBC, in good agreement with theoretical O3LYP/Alhrichs-pVTZ calculations. The 5-hydroxymethyl derivatives (11, 15, 19) showed a (3)J(H, H) coupling constant of methylene protons evidencing the presence of a seven-membered intramolecular hydrogen bonded ring, not observed for the 8-hydroxymethyl isomers.

Coumarins from Galipea panamensis and Their Activity against Leishmania panamensis.

Two new coumarin compounds (1 and 2), phebalosin (3), its derived artifact murralongin (4), and murrangatin acetonide (5) were isolated from the leaves of Galipea panamensis. The structures of 1 and 2 were assigned as 7-{[(2R*)-3,3-dimethyloxiran-2-yl]methoxy}-8-[(2R*,3R*)-3-isopropenyloxiran-2-yl]-2H-chromen-2-one and 7-methoxy-8-(4-methyl-3-furyl)-2H-chromen-2-one, respectively, on the basis of their spectroscopic data (primarily NMR and MS). Compounds 1-3 were tested against axenic amastigote forms of Leishmania panamensis and displayed 50% effective concentrations (EC(50)) of 9.9, 10.5, and 14.1 microg/mL, respectively. These three compounds also displayed cytotoxicity (IC(50)) at concentrations of 9.7, 33.0, and 20.7 microg/mL, respectively, on human promonocytic U-937 cells.

Leishmanicidal activity of aliphatic and aromatic lactones: correlation structure-activity.

Several aliphatic and aromatic lactones and two dimers were synthesized using the sequence: allylation - esterification - metathesis. These compounds were active in vitro against intracellular amastigotes of Leishmania panamensis. The structure-activity relationship showed the importance of the aliphatic side chain to enhance the biological activity and to obtain lower cytotoxicity. It was also observed that a decrease in the size of the lactone ring increases the selectivity index.

Triclosan-caffeic acid hybrids: Synthesis, leishmanicidal, trypanocidal and cytotoxic activities.

The synthesis, cytotoxicity, anti-leishmanial and anti-trypanosomal activities of twelve triclosan-caffeic acid hybrids are described herein. The structure of the synthesized products was elucidated by a combination of spectrometric analyses. The synthesized compounds were evaluated against amastigotes forms of L. (V) panamensis, which is the most prevalent Leishmania species in Colombia, and against Trypanosoma cruzi, which is the pathogenic species to humans. Cytotoxicity was evaluated against human U-937 macrophages. Eight compounds were active against L. (V) panamensis (18-23, 26 and 30) and eight of them against T. cruzi (19-22, 24 and 28-30) with EC50 values lower than 40 µM. Compounds 19-22, 24 and 28-30 showed higher activities than benznidazole (BNZ). Esters 19 and 21 were the most active compounds for both L. (V) panamensis and T. cruzi with 3.82 and 11.65 µM and 8.25 and 8.69 µM, respectively. Compounds 19-22, 24 and 28-30 showed higher activities than benznidazole (BNZ). Most of the compounds showed antiprotozoal activity and with exception of 18, 26 and 28, the remaining compounds were toxic for mammalian cells, yet they have potential to be considered as candidates for anti-trypanosomal and anti-leishmanial drug development. The activity is dependent on the length of the alkyl linker with compound 19, bearing a four-carbon alkyl chain, the most performing hybrid. In general, hydroxyl groups increase both activity and cytotoxicity and the presence of the double bond in the side chain is not decisive for cytotoxicity and anti-protozoal activity.

Ionic liquids intercalated in montmorillonite as the sorptive phase for the extraction of low-polarity organic compounds from water by rotating-disk sorptive extraction.

Montmorillonite (MMT) clays were modified by the intercalation into their galleries of ionic liquids (IL) based on imidazolium quaternary ammonium salts. This new eco-materials exhibited good features for use as a sorptive phase in the extraction of low-polarity analytes from aqueous samples. Spectroscopic analyses of the modified clays were conducted and revealed an increase in the basal spacing and a shifting of the reflection plane towards lower values as a consequence of the effective intercalation of organic cations into the MMT structure. The novel sorbent developed herein was assayed as the sorptive phase in rotating-disk sorptive extraction (RDSE), using polychlorinated biphenyls (PCBs), representative of low-polarity pollutants, as model analytes. The final determination was made by gas chromatography with electron capture detection. Among the synthetized sorptive phases, the selected system for analytical purposes consisted of MMT modified with the 1-hexadecyl-3-methylimidazolium bromide (HDMIM-Br) IL. Satisfactory analytical features were achieved using a sample volume of 5 mL: the relative recoveries from a wastewater sample were higher than 80%, the detection limits were between 3 ng L-1 and 43 ng L-1, the precision (within-run precision) expressed as the relative standard deviation ranged from 2% to 24%, and the enrichment factors ranged between 18 and 28. Using RDSE, the extraction efficiency achieved for the selected MMT-HDMIM-Br phase was compared with other commercial solid phases/supports, such as polypropylene, polypropylene with 1-octanol (as a supported liquid membrane), octadecyl (C18) and octyl (C8), and showed the highest response for all the studied analytes. Under the optimized extraction conditions, this new device was applied in the analysis of the influent of a wastewater treatment plant in Santiago (Chile), demonstrating its applicability through the good recoveries and precision achieved with real samples.

In vitro sensitivity of Botrytis cinerea to anthraquinone and anthrahydroquinone derivatives.

The effect on mycelial growth of the fungus Botrytis cinerea of a set of structurally related tricyclic hydroquinones [9,10-dihydroxy-4,4-dimethyl-2,3,5,8-tetrahydroantracen-1(4H)-one and 9,10-dihydroxy-4,4-dimethyl-5,8-dihydroanthracen-1(4H)-one derivatives] and tricyclic quinones [4,4-dimethylanthracen-1,9,10(4H)-trione derivatives] was studied. In general, the anthraquinones presented higher activity than the anthrahydroquinones. Anthraquinone and anthrahydroquinone derivatives with methyl groups on the A ring showed higher antifungal activity than the unsubstituted ones, 4,4,6,7-tetramethyl-(4H)-anthracene-1,9,10-trione being the most active compound of this set. The presence of a polar group such as hydroxymethyl reduced the activity. The effect of two anthrahydroquinones and two anthraquinones on the conidia germination of the fungus was also determined. Anthrahydroquinones did not affect the germination. The most active compound was 4,4-dimethylanthracene-1,9,10(4H)-trione, with 100% inhibition of germination at 7 h of incubation. These results again suggest that the structure of the anthraquinones is important in exerting an antifungal effect on B. cinerea. Furthermore, possible mechanisms of action of compound 4,4-dimethylanthracene-1,9,10(4H)-trione were studied. This compound did not produce lipoperoxidation of membrane and did not induce the formation of oxygen reactive species, but it was able to permeabilize the plasmatic membrane of B. cinerea, increasing the phosphorus concentration in the intracellular medium.

Synthesis, leishmanicidal, trypanocidal and cytotoxic activity of quinoline-hydrazone hybrids.

Cutaneous leishmaniasis and Chagas disease are vector-borne parasitic disease causing serious risks to million people living in poverty-stricken areas. Both diseases are a major health problem in Latin America, and currently drugs for the effective treatment of these diseases have important concerns related with efficacy or toxicity than need to be addressed. We report herein the synthesis and biological activities (cytotoxicity, leishmanicidal and trypanocidal activities) of ten quinolone-hydrazone hybrids. The structure of the products was elucidated by spectrometric analyses. The synthesized compounds were evaluated against amastigotes forms of L. (V) panamensis which is the most prevalent Leishmania species in Colombia and Trypanosoma cruzi that is the major pathogenic species to humans; in turn, cytotoxicity was evaluated against human U-937 macrophages. Compounds 6b, 6c and 8 showed activity against L. (V) panamensis with EC50 of 6.5 ± 0.8 µg/mL (21.2 µM), 0.8 ± 0.0 µg/mL (2.6 µM) and 3.4 ± 0.6 µg/mL (11.1 µM), respectively, while compounds 6a and 6c had activity against T. cruzi. with EC50 values of 1.4 ± 0.3 µg/mL (4.8 µM) and 6.6 ± 0.3 µg/mL (4.6 µM), respectively. Even compound 6a showed better activity against T. cruzi than the standard drug benznidazole with EC50 = 10.5 ± 1.8 µg/mL (40.3 µM). Analysis of the results obtained against leishmaniasis indicates that antiparasite activity is related to the presence of 2-substituted quinoline (isoquinolinic core) and the hydroxyl group in positions 3 and 4 of the aromatic ring. Although the majority of these compounds were highly cytotoxic, the antiparasite activity was higher than cytotoxicity and therefore, they still have potential to be considered as hit molecules for leishmanicidal and trypanocidal drug development.

Antileishmanial Effect of 5,3'-Hydroxy-7,4'-dimethoxyflavanone of Picramnia gracilis Tul. (Picramniaceae) Fruit: In Vitro and In Vivo Studies.

Species of Picramnia genus are used in folk medicine to treat or prevent skin disorders, but only few species have been studied for biological activity and chemical composition. P. gracilis Tul. is a native species from Central and South America and although its fruits are edible, phytochemical analysis or medicinal uses of this species are not known. In the search of candidates to antileishmanial drugs, this work aimed to evaluate the antileishmanial activity of P. gracilis Tul. in in vitro and in vivo studies. Only ethanolic extract of fruits showed leishmanicidal activity. The majoritarian metabolite was 5,3'-hydroxy-7,4'-dimethoxyflavanone ether that exhibited high activity against L. (V.) panamensis (EC50 17.0 + 2.8 mg/mL, 53.7 µM) and low toxicity on mammalian U-937 cells, with an index of selectivity >11.8. In vivo studies showed that the flavanone administered in solution (2 mg/kg/day) or cream (2%) induces clinical improvement and no toxicity in hamsters with CL. In conclusion, this is the first report about isolation of 5,3'-hydroxy-7,4'-dimethoxyflavanone of P. gracilis Tul. The leishmanicidal activity attributed to this flavanone is also reported for the first time. Finally, the in vitro and in vivo leishmanicidal activity reported here for 5,3'-hydroxy-7,4'-dimethoxyflavanone offers a greater prospect towards antileishmanial drug discovery and development.

Leishmanicidal activity of passifloricin A and derivatives.

Several natural and hemisyntetic passifloricins were assayed against Leishmania panamensis amastigotes. High leishmanicidal activity as well as toxicity was observed. The structure of a new compound is also reported.

Erosive effect of energy drinks alone and mixed with alcohol on human enamel surface: an in vitro study

Objective: to assess the erosive effect of energy drinks (ED) alone and mixed with alcohol on the human enamel surface in vitro. Methods: Twenty non-erupted human third molars were vertically sectioned in half. Specimens were exposed to 5mL of ED plus 5mL of artificial saliva or 5mL of ED plus 5mL of artificial saliva plus 5mL of alcohol (Pisco). Exposure times were set at 30min and 60min. Erosive assessments were made using scanning electron microscopy (SEM) and energy-dispersive x-ray spectroscopy (EDS). The ED analyzed were Mr. Big, Kem Extreme, Red Bull, and Monster Energy. ED pH measurements were performed at 25°C and titration was done with NaOH 0.1mol/L. Results: The pH ranges were: ED alone 2.55 to 3.46, ED mixed with artificial saliva 2.60 to 3.55, ED mixed with Pisco 2.82 to 3.70, and ED mixed with both 2.92 to 3.86. The pH of Pisco was 6.13, and Pisco mixed with artificial saliva had a pH of 6.23. Titration showed a pH range from 3.5 to 5.7. SEM-EDS analysis showed that Mr. Big and Monster led to clear demineralization at 30 min and remineralization at 60m in. Pisco slightly decreased the erosive effect of these ED. Kem Xtreme and Red Bull led to no demineralization in the first hour. Conclusion: According to the pH, acidity and EDS analysis, the ED of the present study likely caused enamel erosion in human teeth surface dependent on exposure time.

Aspectos nutricionales y fisioquímicos de Ilex Laurina Kunth (Aquifoliaceae): un estudio comparativo con Ilex paraguariensis / Nutricional and Physicochemical aspects of Ilex laurina Kunth (Aquifoliaceae): A Comparative Study with Ilex Paraguariensis

Resumen Antecedentes: la Ilex laurina K., planta colombiana cuya infusión presenta actividad antioxidante y antiproliferativa, pertenece al género del té de yerba mate. Objetivo: evaluar las propiedades nutricionales y fisicoquímicas de una infusión de hojas de Ilex laurina K. como potencial materia prima, alimento funcional y sustituyente del convencional té de mate Ilex paraguariensis. Materiales y métodos: análisis por HPLC de fitoquímicos y carbohidratos, solubilidad, medición de minerales por espectrofotometría de absorción atómica y análisis sensorial. Resultados: en la infusión de Ilex laurina K. en comparación con la de Ilex paraguariensis se encontraron mayores concentraciones de ácido clorogénico (429,2±20,2 frente a47,4±1,9 mg/kg), de p-cumárico (47,3±2,4 versus 24,4±0,9 mg/kg), de metilxantinas (1,4-1,8 veces), hierro, cobre y zinc, y similares de ácido cafeico (52,6±18,8 versus 47,4±1,9 mg/L) (p>0,05). Se detectaron ácido ferúlico (21,5±1,6 mg/L) y fructosa (0,4±0,003 g/L), mientras en la de Ilex paraguariensis se encontró glucosa (0,8±0,007 g/L) y mayores concentraciones de magnesio y manganeso (p<0,05). La infusión de Ilex laurina K. fue aceptada por el 64 % de los consumidores, independiente del sexo; así como el color (51 %), olor (47 %), sabor (42 %) y dulzor (47 %). Conclusiones: la infusión de Ilex laurina K. además de tener buena aceptación contiene mayor cantidad de compuestos fenólicos, metilxantinas, hierro, cobre, zinc y fructosa que la de Ilex paraguariensis; y menor aporte de glucosa, magnesio y manganeso.

Abstract Background: Ilex laurina K., a Colombian plant whose infusion presents antioxidant and antiproliferative properties, belongs to a species of Yerba Mate tea. Objective: Evaluate the nutritional and physiochemical properties of an infusion using Ilex laurina K. leaves as a potential raw material, functional food and substitute for the conventional Ilex paraguariensis tea. Objective: To evaluate the nutritional and physiochemical properties of an infusion using Ilex laurina K. leaves as a potential raw material, functional food and substitute for the conventional Ilex paraguariensis tea. Materials and Methods: Phytochemicals and carbohydrates by HPLC analysis, solubility, mineral measurement using atomic absorption spectrophotometry, and sensorial analysis. Results: The Ilex laurina K. infusión compared with Ilex paraguariensis infusion found higher concentrations of chlorogenic acid (429.2 ± 20.2 versus 47.4 ± 1.9 mg / kg), p-coumaric (47.3 ± 2.4 versus 24.4 ± 0.9 mg / kg), xanthines (1.4-1.8 times), iron, copper, zinc, and the likes of caffeic acid (52.6 ± 18.8 versus 47.4 ± 1.9 mg / L) (p> 0.05). Ferulic acid (21.5 ± 1.6 mg / L) and fructose (0.4 ± 0.003 g / L) were detected, while glucose (0.8 ± 0.007 g / L) and higher concentrations of magnesium and manganese (p<0.05) we found in the Ilex paraguariensis. Ilex laurina K. infusion was accepted by 64% of consumers, regardless of gender; and respective acceptability of the following characteristics: color (51%), smell (47%), flavor (42%) and sweetness (47%). Conclusions: Ilex laurina K. infusion is well accepted by consumers, and contains a higher concentration of phenol compounds, xanthines, iron, copper, zinc and fructose than Ilex paraguariensis infusion, and contributes less glucose, magnesium and manganese.

Effects of 9,10-dihydroxy-4,4-dimethyl-5,8-dihydro-1(4H)-anthracenone derivatives on tumor cell respiration.

A series of tricyclic hydroquinones, incorporating a carbonyl group in the ortho position relative to the phenol function, were tested as inhibitors of oxygen uptake against the TA3 mouse carcinoma cell line and its multidrug-resistant variant TA3-MTX-R. The title compound, which proved to be the most active one, also exhibited low micromolar dose-dependent growth inhibition of the human tumor U937 cell line (human monocytic leukemia). A tentative structure-activity relationship is proposed for these substances. A comparison between the cytotoxicities of the title compound and 4,4-dimethyl-5,8-dihydroxynaphthalene-1-one, with their activities as inhibitors of oxygen uptake by the TA3-MTX-R cell line, is presented. Also, the inhibition of oxygen uptake by 6-(4-methylpent-3-enyl)-1,4-naphthoquinone was determined and compared with its reported cytotoxicity toward P-388 (murine lymphocytic leukemia), A-549 (human lung carcinoma), HT-29 (human colon carcinoma), and MEL-28 (human melanoma) cells. The inhibition of oxygen uptake by TA3-MTX-R cells is useful as a quick test for preliminary screening of possible anticancer activity.