Effect of chito-oligosaccharide on the intestinal absorptions of phenylethanoid glycosides in Fructus Forsythiae extract

Phenylethanoid glycosides, the main active ingredients in Fructus Forsythiae extract possesses strong antibacterial, antioxidant and antiviral effects, and their contents were higher largely than that of other ingredients such as lignans and flavones, but their absolute bioavailability orally was significantly low, which influenced clinical efficacies of its oral preparations seriously. In the present study, the absorption mechanism of phenylethanoid glycosides was studied using in vitro Caco-2 cell model. And the effect of chito-oligosaccharide (COS) on the intestinal absorption of phenylethanoid glycosides in Fructus Forsythiae extract was investigated using in vitro, in situ and in vivo models. The pharmacological effects such as antiviral activity improvement by COS were verified by MDCK cell damage inhibition rate after influenza virus propagation. The observations from in vitro Caco-2 cell showed that the absorption of phenylethanoid glycosides in Fructus Forsythiae extract so with that in monomers was mainly restricted by the tight junctions, and influenced by efflux transporters (P-gp and MRP2). Meanwhile, the absorption of phenylethanoid glycosides in Fructus Forsythiae extract could be improved by COS. Besides, COS at the same low, medium and high concentrations caused a significant, concentration-dependent increase in the P_app-value for phenylethanoid glycosides compared to the control group (p < 0.05), and was all safe for the Caco-2 cells. The observations from single-pass intestinal perfusion in situ model showed that the intestinal absorption of phenylethanoid glycosides can be enhanced by COS. Meanwhile, the absorption enhancing effect of phenylethanoid glycosides might be saturable in different intestine sites. In pharmacokinetics study, COS at dosage of 25 mg/kg improved the bioavailability of phenylethanoid glycosides in Fructus Forsythiae extract to the greatest extent, and was safe for gastrointestine from morphological observation. In addition, treatment with Fructus Forsythiae extract with COS at dosage of 25 mg/kg prevented MDCK cell damage upon influenza virus propagation better than that of control. All findings above suggested that COS at dosage of 25 mg/kg might be safe and effective absorption enhancer for improving the bioavailability of phenylethanoid glycosides and the antiviral activity in vitro in Fructus Forsythiae extract.     

Synthesis and pharmacological evaluation of nucleoside prodrugs designed to target siderophore biosynthesis in Mycobacterium tuberculosis

The nucleoside antibiotic, 5′-O-[N-(salicyl)sulfamoyl]adenosine (1), possesses potent whole-cell activity against Mycobacterium tuberculosis (Mtb), the etiological agent of tuberculosis (TB). This compound is also active in vivo, but suffers from poor drug disposition properties that result in poor bioavailability and rapid clearance. The synthesis and evaluation of a systematic series of lipophilic ester prodrugs containing linear and α-branched alkanoyl groups from two to twelve carbons at the 3′-position of a 2′-fluorinated analog of 1 is reported with the goal to improve oral bioavailability. The prodrugs were stable in simulated gastric fluid (pH 1.2) and under physiological conditions (pH 7.4). The prodrugs were also remarkably stable in mouse, rat, and human serum (relative serum stability: human ∼ rat ≫ mouse) displaying a parabolic trend in the SAR with hydrolysis rates increasing with chain length up to eight carbons (t_1/2 = 1.6 h for octanoyl prodrug 7 in mouse serum) and then decreasing again with higher chain lengths. The permeability of the prodrugs was also assessed in a Caco-2 cell transwell model. All of the prodrugs were found to have reduced permeation in the apical-to-basolateral direction and enhanced permeation in the basolateral-to-apical direction relative to the parent compound 2, resulting in efflux ratios 5–28 times greater than 2. Additionally, Caco-2 cells were found to hydrolyze the prodrugs with SAR mirroring the serum stability results and a preference for hydrolysis on the apical side. Taken together, these results suggest that the described prodrug strategy will lead to lower than expected oral bioavailability of 2 and highlight the contribution of intestinal esterases for prodrug hydrolysis.     

Optimization of extraction method for LC–MS based determination of phenolic acid profiles in different Impatiens species

The main aim of presented study was the comparison of various extraction methods for the quantitative and qualitative analysis (LC-ESI–MS/MS) of phenolic acids present in extracts obtained from leaves, flowers, and roots of Impatiens glandulifera. The accelerated solvent extraction (ASE) at three temperature ranges (80° C, 100° C, and 120° C), ultrasound assisted extraction (USAE) at 60° C, and traditional extraction in Soxhlet apparatus were used. Taking into account the extraction yield, and the diversity of the individual compounds, ultrasound assisted extraction proved to be the most efficient method, and it was used to determine the content of phenolic acids in leaves of four other Impatiens species, including I. balsamina, I. noli-tangere, I. parviflora, and I. walleriana. Eleven phenolic acids were identified in all examined species. These were protocatechuic, gentisic, 4- hydroxybenzoic, vanillic, trans-caffeic, syringic, trans-p-coumaric, trans- and cis-ferulic, salicylic, and 3-hydroxycinnamic acids. In the extract from the leaves of I. balsamina and I. walleriana, gallic and cis-p-coumaric acids were found additionally. The most abundant compounds in all examined extracts were protocatechuic and 3-hydroxycinnamic acids. The latest acid was found in the highest yield in I. noli-tangere (266.12 μg/g DW). In the leaves of I. glandulifera a great amount of 4-hydroxybenzoic (41.44 μg/g DW), vanillic (61.50 μg/g DW), and trans-p-coumaric (58.42 μg/g DW) acids was also observed. Our results indicate that protocatechuic, 4-hydroxybenzoic, vanillic, trans-p-coumaric, trans-ferulic, and 3-hydroxycinnamic acids were most characteristic of Impatiens species.Additionally, various phenolic-rich extracts from leaves, flowers, and roots of Impatiens glandulifera were tested for antioxidant activity. The highest antiradical activity was detected for roots using Soxhlet extraction (EC50 = 0.055 mg [DE/ml]).The study demonstrated that members of the genus Impatiens, and in particular Impatiens glandulifera, and Impatiens noli-tangere, contain significant amounts of phenolic acids. In addition, extracts from various parts of I. glandulifera could be interesting as novel sources of natural antioxidants.     

Studies on the accumulation of phenolic acids and flavonoids in different in vitro culture systems of Schisandra chinensis (Turcz.) Baill. using a DAD-HPLC method

Different in vitro culture systems of the East-Asian origin medicinal plant species − Schisandra chinensis, were tested in order to investigate their potential for the accumulation of two groups of phenolic compounds. In vitro cultures were maintained on the Murashige and Skoog (MS) medium supplemented with 3 mg/l BA and 1 mg/l NAA in an agar system (30- and 60-day growth cycles), and also in two different liquid systems: stationary and agitated. Stationary liquid cultures were grown in batch (30- and 60-day growth cycles) and fed-batch modes. Of the twenty compounds, seven free phenolic acids and of the eleven compounds, five flavonoids were quantified in methanolic extracts from lyophilized biomass and in the growth media using the RP-HPLC-DAD method. For comparison purposes, phytochemical analyses of leaf and fruit extracts from the parent plant were also conducted. The estimated compounds were not detected in the growth media. The highest total amounts of phenolic acids (71.48 mg/100 g DW) and flavonoids (29.36 mg/100 g DW) were found in extracts from the biomass of agar cultures harvested after 30 days of cultivation. The main metabolites in all the tested systems were: protocatechuic acid (max. 35.69 mg/100 g DW), chlorogenic acid (max. 13.05 mg/100 g DW), and quercitrin (max. 27.43 mg/100 g DW).     

Filamentous fungi from Plantago lanceolata L. leaves: Contribution to the pattern and stability of bioactive metabolites

The aim of this study was to test contribution of plant-associated microorganism (PAMs) to metabolite stability/instability in a medicinal plant matrix.Therefore, PAM strains were isolated and identified based on relevant DNA sequences from Plantago lanceolata leaves. Sterile water extracts of P. lanceolata were incubated with the isolated strains and antioxidants (ascorbic acid (AA), and EDTA) for 15 days, and changes in the concentrations of chief bioactive constituents (aucubin, catalpol, acteoside (=verbascoside)) were quantified by capillary electrophoresis. Phenolic breakdown-products were identified by GC–MS.PAMs were identified from the genera Epicoccum, Bipolaris, Cladosporium, Leptosphaerulina, Aspergillus, Eurotium and Penicillium (pathongens, endophytes, and other species). Some fungi caused significant decomposition of the chief constituents (p < 0.001). Surprisingly, some strains inhibited breakdown of acteoside (p < 0.001). Meanwhile, concentration of several phenolic acids increased in fungi-infested extracts (p < 0.001). Gentisic acid, 4-hydroxyphenyl acetic acid, 4-hydroxybenzoic acid and hydroxytyrosol were only present when the extract was infested with a PAM. The products are powerful antioxidants and chelators. Concentrations of phenolic acids influenced acteoside stability significantly (p < 0.01), as shown by basic data-mining techniques. AA and EDTA also significantly inhibited acteoside breakdown in sterile model solutions (p < 0.05).Our results suggest that the phenolic acid mixture (produced during the fungal proliferation) protected acteoside from breakdown, possibly via its antioxidant activity and metal complexing ability. It was shown that PAMs can increase or decrease the stability of chief metabolites in herbal matrices, and can significantly alter the chemical pattern of the plant matrix.     

d-Xylose absorption test: A tool for the assessment of the effect of anticoccidials on the intestinal absorptive capacity of broilers during experimental coccidiosis

Coccidiosis in chickens causes intestinal mucosal lesions and disrupts its integrity leading to a disturbance in absorption of dietary components. The d-xylose absorption test is a sensitive tool of measuring the absorption capacity of the intestine in diseased chickens. In an experiment on broilers, the influence of different anticoccidials on the intestinal absorption capacity of the birds challenged with experimental coccidiosis was evaluated, using the d-xylose absorption test. The experiment had 5 groups of 10 Ross male broiler chickens (24-days-old) as follows: Group 1— negative control received no Eimeria oocystes, Group 2—positive control challenged with mixed Eimeria oocystes, Group 3—positive control dosed with an attenuated oral coccidiosis vaccine, Group 4—positive control dosed with 25 ppm toltrazuril in drinking water and Group 5—positive control received 66 ppm salinomycin sodium, in the diet. The d-xylose absorption test was carried out 5 days after the coccidial infection. Results showed that coccidiosis highly reduced the plasma d-xylose peak level of Group 2 when compared with Group 1 (31 mg/dl at 90 min versus 50 mg/dl at 30–60 min after the d-xylose administration, respectively). The concentration of d-xylose followed cubic (P<0.001, r² = 0.886) and quadratic (P<0.001, r² = 0.686) correlations with time in Group 1 and 2, respectively. Anticoccidials enhanced the uptake of d-xylose in the infected birds. The plasma d-xylose reached to its peak in Group 3, 4, and 5 (38.9, 50 and 47.0 mg/dl, respectively) at 60–90 min after the d-xylose administration and had quadratic functions with time (r² = 0.802, 0.883 and 0.860, respectively, P<0.001). The d-xylose absorption test was a sensitive test for evaluating the influence of anticoccidials on the absorption capacity of intestinal mucosae during coccidiosis in broiler chickens.     

Effects of l-glutamine supplementation alone or with antioxidants on hydrogen peroxide-induced injury in human intestinal epithelial cells

Background and aimsIn situations of oxidative stress, l-glutamine (Gln) exhibits protective effects which may be potentiated by its combination with antioxidants. The aim of this study was to compare the effects of a Gln-antioxidants formula vs Gln alone in intestinal epithelial cell lines Caco-2 and HCT-8 submitted to hydrogen peroxide-induced oxidative injury.MethodsCells were cultured during 24 h with 0, 1, 2, 4, 8 or 16 mM Gln or isomolar Gln combined to cysteine, vitamine C, vitamine E, β-carotene, Zn and Se (Gln-AOX). After 24 h, membrane integrity was assessed by means of LDH leakage, the level of oxidative stress by analysis of 8-isoprostane concentration and cell viability by colorimetric-MTT assay.ResultsLDH activity decreased (P < 0,05) with a dose-dependent manner in both cell types with Gln-AOX whereas Gln decreased LDH release only in the Caco-2 line at 1 mM. For both cell lines, release of 8-isoprostane was not blunted by any treatment and increased paradoxically with 16 mM Gln (P < 0.05). Cell viability was higher (P < 0.05) using Gln-AOX vs Gln at 4, 8 and 16 mM in both cell lines.ConclusionsThese results suggest that Gln-AOX is more efficient than Gln alone to preserve cell membrane integrity and viability in intestinal cell lines submitted to oxidative injury.     

Antiviral efficacy of Limonium densiflorum against HSV-1 and influenza viruses

Viral infections remain a major threat to humans and animals and there is a crucial need for new antiviral agents especially with the development of resistant viruses. Several Limonium genus members (Plumbaginacea) have been widely used in traditional medicine for the treatment of infections. In this study, we investigated the antiviral activities of different fractions after successive extraction (hexane, dichloromethane, ethanol and methanol) of the halophyte Limonium densiflorum against H1N1 influenza and HSV-1 herpes viruses. In addition, TLC phytochemicals of the shoot extracts were analyzed. All extracts were tested for their cytotoxicity using a fluorometric resazurin assay. The antiviral activity of extracts was tested using four modes of action: virucidal test, pretreatment of cells with samples before infection, attachment assay and plaque reduction test. A good antiviral activity was found with ethanol and methanol extracts. They were most potent in HSV-1 inhibition than H1N1 influenza virus. The most potent inhibition was observed with ethanol extract, and it exhibited high levels of virucidal activity against HSV-1 (IC₅₀ = 6 μg/mL). It inhibits the replication of the virus by 75% when added after penetration of the virus, and by 100% when added during the viral attachment. It protects MDCK cells against influenza virus by abolishing virus to entry into the host cell (IC₅₀ = 55 μg/mL). After attachment of influenza virus, the ethanol extract displayed an appreciable inhibition of virus replication (IC₅₀ = 193 μg/mL). Methanol extract showed a moderate antiviral capacity against both viruses. While dichloromethane has excellent antiherpes potential, results were inappropriate because it was toxic to Vero cells, hexane extract has no effect. TLC analysis of these extracts showed that flavonoids and saponins were the major classes of natural products found in the shoot extracts that may be responsible for these antiviral activities.     

Antioxidative,antimicrobial and cytotoxic effects of the phenolics of Leea indica leaf extract

This study investigated the phytochemical, antioxidative, antimicrobial and cytotoxic effects of Leea indica leaf ethanol extract. Phytochemical values namely total phenolic and flavonoid contents, total antioxidant capacity, DPPH radical scavenging effect, FeCl₃ reducing power, DMSO superoxide scavenging effect and Iron chelating effects were studied by established methods. Antibacterial, antifungal and cytotoxic effects were screened by disk diffusion technique, food poison technique and brine shrimp bioassay, respectively. Results showed the total phenolic content 24.00 ± 0.81 g GAE/100 g, total flavonoid content 194.68 ± 2.43 g quercetin/100 g and total antioxidant capacity 106.61 ± 1.84 g AA/100 g dry extract. Significant (P < 0.05) IC₅₀ values compared to respective standards were recorded in DPPH radical scavenging (139.83 ± 1.40 μg/ml), FeCl₃ reduction (16.48 ± 0.64 μg/ml), DMSO superoxide scavenging (676.08 ± 5.80 μg/ml) and Iron chelating (519.33 ± 16.96 μg/ml) methods. In antibacterial screening, the extract showed significant (P < 0.05) zone of inhibitions compared to positive controls Ampicillin and Tetracycline against Gram positive Bacillus subtilis, Bacillus cereus, Bacillus megaterium, and Staphylococcus aureus and Gram negative Salmonella typhi, Salmonella paratyphi, Pseudomonas aeroginosa, Shigella dysenteriae, Vibrio cholerae, and Escherichia coli. Significant minimum inhibitory concentrations compared to tetracycline were obtained against the above organisms. In antifungal assay, the extract inhibited the growth of Aspergillus flavus, Candida albicans and Fusarium equisetii by 38.09 ± 0.59, 22.58 ± 2.22, and 22.58 ± 2.22%, respectively. The extract showed a significant LC₅₀ value compared to vincristine sulfate in cytotoxic assay. The results evidenced the potential antioxidative, antimicrobial and cytotoxic capacities of Leea inidica leaf extract to be processed for pharmaceutical use.     

Identification of serotonin 5-HT1A receptor partial agonists in ginger

Animal studies suggest that ginger (Zingiber officinale Roscoe) reduces anxiety. In this study, bioactivity-guided fractionation of a ginger extract identified nine compounds that interact with the human serotonin 5-HT_1A receptor with significant to moderate binding affinities (K_i = 3–20 μΜ). [³⁵S]-GTPγS assays indicated that 10-shogaol, 1-dehydro-6-gingerdione, and particularly the whole lipophilic ginger extract (K_i = 11.6 μg/ml) partially activate the 5-HT_1A receptor (20–60% of maximal activation). In addition, the intestinal absorption of gingerols and shogaols was simulated and their interactions with P-glycoprotein were measured, suggesting a favourable pharmacokinetic profile for the 5-HT_1A active compounds.     

Antimicrobial activity of medicinal plants and induction of defense related compounds in banana fruits cv. Robusta against crown rot pathogens

A total of 72 plant extracts were tested in vitro for their ability to inhibit the mycelial growth of Lasiodiplodia theobromae and Colletotrichum musae the causal agents of crown rot disease of banana. The results showed that the leaf extract of Zimmu (an interspecific hybrid of Allium cepa L. × Allium sativum L.) and tuber extract of Zehneria scabra recorded maximum inhibition of mycelial growth and spore germination of both the test pathogens. The dipping of banana fruits in Zimmu leaf extract at 25% conc. exhibited 100% inhibition of crown rot disease in cold storage (14 °C) up to 35 days and increased the shelf life to 64 days. However, at room storage (28 ± 2 °C), the same treatment exhibited 86% inhibition of crown rot disease up to 12 days. It was found that the treatment of banana fruits with Zimmu leaf extract did not alter the organoleptic properties of banana. The biochemical analysis of banana fruits treated with Zimmu leaf extract showed significant increase in phenylalanine ammonia-lyase (PAL), chitinase and β-1,3-glucanase activities and enhanced accumulation of phenolic compounds compared to other treatments. These findings suggest that the effect of Zimmu leaf extract on crown rot disease may be associated with the direct fungi toxic property against the test pathogens and elicitation of defense related compounds in banana fruits.     

Kiwifruit cysteine protease actinidin compromises the intestinal barrier by disrupting tight junctions

BackgroundThe intestinal epithelium forms a barrier that food allergens must cross in order to induce sensitization. The aim of this study was to evaluate the impact of the plant-derived food cysteine protease — actinidin (Act d1) on the integrity of intestinal epithelium tight junctions (TJs).MethodsEffects of Act d1 on the intestinal epithelium were evaluated in Caco-2 monolayers and in a mouse model by measuring transepithelial resistance and in vivo permeability. Integrity of the tight junctions was analyzed by confocal microscopy. Proteolysis of TJ protein occludin was evaluated by mass spectrometry.ResultsActinidin (1 mg/mL) reduced the transepithelial resistance of the cell monolayer by 18.1% (after 1 h) and 25.6% (after 4 h). This loss of barrier function was associated with Act d 1 disruption of the occludin and zonula occludens (ZO)-1 network. The effect on intestinal permeability in vivo was demonstrated by the significantly higher concentration of 40 kDa FITC-dextran (2.33 μg/mL) that passed from the intestine into the serum of Act d1 treated mice in comparison to the control group (0.5 μg/mL). Human occludin was fragmented, and putative Act d1 cleavage sites were identified in extracellular loops of human occludin.ConclusionAct d1 caused protease-dependent disruption of tight junctions in confluent Caco-2 cells and increased intestinal permeability in mice.General significanceIn line with the observed effects of food cysteine proteases in occupational allergy, these results suggest that disruption of tight junctions by food cysteine proteases may contribute to the process of sensitization in food allergy.     

Phenolic compounds composition and physiological attributes of Matricaria chamomilla grown in copper excess

Four-week old plants of chamomile (Matricaria chamomilla) cultivated in nutrient solution were exposed to copper (3, 60 and 120 μM) for 10 days. At 120 μM, Cu decreased dry mass production, water, chlorophyll and nitrogen content in both the leaf rosettes and roots. Five phenolic acids were detected in methanol extracts of the leaf rosettes (protocatechuic, p-hydroxybenzoic, vanillic, chlorogenic and salicylic acid) and six additional compounds (gentisic, syringic, caffeic, sinapic and o-/p-coumaric acid) were released after acid hydrolysis. Most of the 11 phenolic acids detected increased in 60 μM Cu but in the 120 μM treatment their contents were lower or not significantly different from the control. Among coumarin-related compounds, (Z)- and (E)-2-ß-d-glucopyranosyloxy-4-methoxycinnamic acids increased in 60 and 120 μM Cu while herniarin rose in the 3 and 60 μM Cu by the end of the experiment. The amounts of umbelliferone were not affected by any of the doses tested. These facts in relation to antioxidative properties of phenolic metabolites are also discussed. The malondialdehyde content of the leaf rosettes was not affected by exposure of plants to 120 μM Cu in a time-course experiment but in the roots a sharp increase was observed after 24 and 48 h of treatment. At 120 μM, Cu stimulated a 9-fold higher K⁺ loss than the 60 μM treatment while at the lowest concentration it stimulated potassium uptake. Cu accumulation in the roots was 3-, 49- and 71-fold higher than that in the leaf rosettes in the 3, 60, and 120 μM Cu treatments, respectively. Results suggest that 120 μM Cu dose is limiting for chamomile growth under the conditions of present research.     

Chemistry,antioxidant and antimicrobial potential of nutmeg (Myristica fragrans Houtt)

Antioxidant and antimicrobial activities of nutmeg (Myristica fragrans Houtt) seed extracts were evaluated. Seeds were extracted with acetone, ethanol, methanol, butanol and water. All the extracts have shown significant antioxidant and antimicrobial activities against the tested microorganisms. Among all extracts, acetone extract has shown the highest antioxidant activity. The acetone extract showed 93.12 ± 1.48 mg gallic acid equivalents (GAE)/100 g dry weight total phenolic content, DPPH scavenging activity of 63.04 ± 1.56%, chelating activity of 64.11 ± 2.21% and 74.36 ± 1.94% inhibition of β-carotene bleaching, at 1 mg/mL extract concentration. Out of all extracts, acetone extract was able to exert antimicrobial activity against all tested bacteria and fungi. Acetone extract has shown the strongest antibacterial and antifungal activity with Staphylococcus aureus (13.8 ± 0.42 mm) and Aspergillus niger (14.4 ± 0.37 mm), respectively. GC–MS analysis of acetone extract has revealed the presence of 32 compounds of extract representing 99.49%. Sabinene (28.61%) has shown the highest occurrence in the extract. β-Pinene (10.26), α-pinene (9.72), myristicin (4.30%), isoeugenol (2.72%), p-cymene (1.81%), carvacrol (1.54%), eugenol (0.89%) and β-caryophellene (0.82%) were reported as possible contributor for antioxidant and antimicrobial activity of nutmeg.     

Altitudinal differences in the contents of phenolics in flowering heads of three members of the tribe Lactuceae (Asteraceae) occurring as introduced species in New Zealand

Crepis capillaris, Hieracium pilosella, and Hypochaeris radicata were investigated for the influence of the altitude of the collection site on the content of phenolics within the flowering heads. These three taxa from the Lactuceae tribe of the Asteraceae family originate from Europe and are now widespread within New Zealand. Flowering heads collected from different altitudes ranging from 180 m to 1060 m (C. capillaris), from 190 to 1290 m (H. pilosella), and from 20 m to 1290 m (H. radicata), respectively, were extracted and analysed by high performance liquid chromatography. Results showed a positive correlation between the altitude of the growing site and the contents of flavonoids and phenolic acids for all investigated taxa. The altitudinal effect was, however, partially concealed by geographic differences between coastal and inland collection sites, with the inland collections containing higher concentrations of flavonoids and phenolic acids than plants collected from the coast. The results are discussed in the light of a putative UV-B protective function of the quantified compounds and of the immigration histories of the three species at hand.     

Free fatty acid receptor 1 (GPR40) agonists containing spirocyclic periphery inspired by LY2881835

The free fatty acid receptor 1 (FFA1), a G protein-coupled receptor (GPCR) naturally activated by long-chain fatty acids is a novel target for the treatment of metabolic diseases. The basic amine spirocyclic periphery of Eli Lilly’s drug candidate LY2881835 for treatment of type 2 diabetes mellitus (which reached phase I clinical trials) inspired a series of novel FFA1 agonists. These were designed to incorporate the 3-[4-(benzyloxy)phenyl]propanoic acid pharmacophore core decorated with a range of spirocyclic motifs. The latter were prepared via the Prins cyclization and subsequent modification of the 4-hydroxytetrahydropyran moiety in the Prins product. Here, we synthesize 19 compounds and test for FFA1 activity. Within this pilot set, a nanomolar potency (EC₅₀ = 55 nM) was reached. Four lead compounds (EC₅₀ range 55–410 nM) were characterized for aqueous solubility, metabolic stability, plasma protein binding and Caco-2 permeability. While some instability in the presence of mouse liver microsomes was noted, mouse pharmacokinetic profile of the compound having the best overall ADME properties was evaluated to reveal acceptable bioavailability (F = 10.3%) and plasma levels achieved on oral administration.     

Free radical scavenging activity from different extracts of leaves of Bauhinia vahlii Wight & Arn.

The objectives of this study were to determine phenolic content and antioxidant activities of chloroform, acetone, methanol and hot water extracts of Bauhinia vahlii leaves. The hot water extract afforded the highest yield (6.3%) while the lowest yield was obtained from the chloroform extract (2.1%). The methanol extract contains higher levels of total phenolics (48.7 ± 0.7 g GAE/100 g extract), tannins (21.7 ± 0.7 g GAE/100 g extract) and flavonoids (10.3 ± 0.2 RE/100 g extract). The extracts were subjected to assess their antioxidant potential using various in vitro systems such as DPPH, ABTS⁺, FRAP, OH, β-carotene linoleic acid bleaching system, phosphomolybdenum reduction and Fe²⁺ chelation. It is concluded that the methanolic extract of B. vahlii leaves have strong antioxidant potential. Further study is necessary for isolation and characterization of the active antioxidants, which may serve as a potential source of natural antioxidants.     

Pancreatic α-amylase and lipase inhibitory activity of polyphenolic compounds present in the extract of black chokeberry (Aronia melanocarpa L.)

The aim of this study was to investigate the effect of black chokeberry (Aronia melanocarpa L.) extract on the activity of porcine pancreatic α-amylase and lipase. An in vitro study demonstrated that three kinds of chokeberry extracts: methanolic, water and acetic caused inhibition of α-amylase and lipase. The methanolic and acetic extracts exhibited the highest inhibitory activities against α-amylase with the IC₅₀ values of 10.31 ± 0.04 mg/ml and pancreatic lipase 83.45 ± 0.50 mg/ml, respectively. In order to identify the compounds which may be the potential inhibitors of α-amylase and lipase, chokeberry extract was analyzed by preparative reverse phase chromatography and high performance liquid chromatography–mass spectrometry (HPLC–MS). These studies have shown that both anthocyanins and phenolic acids are compounds which inhibit the ability of the reaction catalyzed by α-amylase and lipase. The most effective inhibitor of pancreatic α-amylase was chlorogenic acid (IC₅₀ = 0.57 ± 0.16 mg/ml). In the group of anthocyanins the most potent inhibitor of α-amylase was cyanidin-3-glucoside (IC₅₀ = 1.74 ± 0.04 mg/ml), which also showed an ability to inhibit the reaction catalyzed by pancreatic lipase (IC₅₀ = 1.17 ± 0.05 mg/ml). These findings seem to indicate the use of chokeberry as a functional food component, contributing to its anti-obesity activities.     

Straightforward entry to pyrido[2,3-d]pyrimidine-2,4(1H,3H)-diones and their ADME properties

A straightforward synthesis of pyrido[2,3-d]pyrimidine-2,4(1H,3H)-diones was developed starting from 2-chloropyridine-3-carboxylic acid by esterification, nucleophilic aromatic substitution and amide formation in one step, and ring closure allowing their synthesis with two identical or two different group attached to nitrogen. The structural diversity of these [2,3-d]pyrimidine-2,4(1H,3H)-diones resulted in significant variation in the biopharmaceutical properties. This was reflected by the broad range in fasted state simulated intestinal fluid solubility values (12.6 μM to 13.8 mM), Caco-2 permeability coefficients (1.2 × 10⁻⁶ cm/s to 90.7 × 10⁻⁶ cm/s) and in vitro-predicted human in vivo intrinsic clearance values (0 to 159 ml/min/kg).