Galloyl Derivatives Research Articles

Species swarms and their caterpillar colonisers: phylogeny and polyphenols determine host plant specificity in New Guinean Lepidoptera

The majority of multi-cellular terrestrial life is found in tropical forests and is either an invertebrate or a plant: for decades ecologists have sought to understand why. As global change erodes the list of extant species on our planet quantifying what species remain, along with their origins and ecology, contributes to our ability to preserve ecosystem functioning and resilience. Here we study three feeding guilds of caterpillars (Lepidoptera) and seek to understand the drivers of their diet breadth across four diverse tropical plant genera in Papua New Guinea. Host specificity is central to biodiversity estimates and the resilience of ecological networks. Specifically, we calculate distance-based host specificity in relation to plant phylogenetic relationships alongside chemical and mechanical traits of leaves. In terms of chemical defenses, we focus on the major polyphenol groups, a compound class shared across many plant species. We refine our data exploration using food webs and ordinations to pick out specific traits of relevance to insect host specificity. Our results showed that the degree of specialization for caterpillars took the following order: phylogenetic>polyphenol>mechanical, such that insect specificity was explained best by host phylogeny and polyphenol chemistry in our study system. Leaf mining insects had higher host specificity than those feeding externally. Of the traits studied hexahydroxydiphenoyl derivatives, galloyl derivatives, trichome density, quinic acid derivatives, myricetins and successional index explained the most variation in overall insect community structure. Our findings build on earlier studies of New Guinean rainforest communities and add a mechanistic explanation to previous findings that host genera are functional islands for insect herbivores. Further, we demonstrate that different plant genera combine different defensive traits that appear to drive associated insect diversity. Our approach integrates trait data and phylogeny to explore dimensions of specialization and we welcome metabolomic studies that will provide more detailed explanations for insect-herbivore host use. Finally, chemical diversity is directly linked to organismal diversity and by studying a range of insect herbivore guilds we make a connection between feeding ecology and specialization that will help to predict species interactions and, potentially, the persistence of ecological networks.

Colon-available mango (poly)phenols exhibit mitigating effects on the intestinal barrier function in human intestinal cell monolayers under inflammatory conditions.

This study investigated the impact of in vivo available colon-mango (poly)phenols on stress-induced impairment of intestinal barrier function. Caco-2/HT29-MTX cells were incubated with six extracts of ileal fluid collected pre- and 4-8 h post-mango consumption before being subjected to inflammatory stress. (Poly)phenols in ileal fluids were analysed by UHPLC-HR-MS. Epithelial barrier function was monitored by measurement of trans-epithelial electrical resistance (TEER) and the production of selected inflammatory markers (interleukin-8 (IL-8) and nitric oxide (NO)) and the major mucin of the mucosal layer (MUC2). Post-mango intake ileal fluids contained principally benzoic acids, hydroxybenzenes and galloyl derivatives. There was a high interindividual variability in the levels of these compounds, which was reflected by the degree of variability in the protective effects of individual ileal extracts on inflammatory changes in the treated cell cultures. The 24 h treatment with non-cytotoxic doses of extracts of 4-8 h post-mango intake ileal fluid significantly reduced the TEER decrease in monolayers treated with the inflammatory cytomix. This effect was not associated with changes in IL-8 expression and secretion or claudine-7 expression. The mango derived-ileal fluid extract (IFE) also mitigated cytomix-dependent nitrite secretion, as a proxy of NO production, and the MUC2 reduction observed upon the inflammatory challenge. These insights shed light on the potential protective effect of mango (poly)phenols on the intestinal barrier exposed to inflammatory conditions.

Nutritional, chemical and functional potential of Inga laurina (Fabaceae): A barely used edible species

Inga laurina is a plant species which produces edible fruits, and until now there is little information available concerning its nutritional, chemical and bioactive composition. In this study, we evaluated for the first time the proximate composition and mineral contents in its fruit (peel, pulp and seed), that is the traditionally consumed part. The seeds obtained the highest protein (19.52 g/100 g), carbohydrate (22.5 g/100 g) and mineral contents, mainly Cu, Cr, P, Mn, Se and Zn. The peel and pulp were excellent sources of fiber (4.5 and 11.05 g/100 g) as well as mineral content, with Cr and Cu standing out in the pulp. This study is notably the first to provide a detailed assessment of the nutritional compositions of traditionally consumed and not consumed parts of this fruit. Sensory analysis of the pulp was also performed, which indicated good acceptance. The antioxidant properties were characterized in the fruit, peels and leaves. The ABTS test showed that leaf supernatant hydroethanolic crude extract (EC50 = 2.70 μg/mL) and its corresponding ethyl acetate (EC50 = 1.68 µg/mL) and butanol (EC50 = 2.48 µg/mL) partitions presented higher antioxidant potential compared to the control Ginkgo biloba (EC50 = 12.17 µg/mL). The most active precipitate extract regarding DPPH was from the peel (EC50 = 13.30 μg /mL) and the most active partition was the ethyl acetate (EC50 = 13.37 μg/mL), both with better activity compared to the control Ginkgo biloba (EC50 = 46.97 μg/mL). The ethyl acetate partition (EC50 = 13.45 μg/mL) and butanol partition (EC50 = 7.97 μg/mL) from the leaves showed the highest antioxidant capacity. Thus, extracts and partitions from the peels and leaves were studied from a phytochemical point of view due to presenting the best results for antioxidant capacity. The presence of phenolic compounds such as myricetin-3-O-rhamnopyranoside, myricetin-3-O-(2″-O-galloyl)-rhamnopyranoside and myricetin-3-O-(2″,4″-di-O-galloyl)-arabinopentoside-methyl ether were observed in the leaf crude extract and polar partitions, being reported for the first time in the Inga genus and Fabaceae family. Moreover, quercetin, quercetin-3-O-galatoctoside, quercetin-3-O-rhamnopyranoside, quercetin-3-O-(2″-O-galloyl)-rhamnoside, and quercetin tri-hexose were identified in the peel crude extract and ethyl acetate partition, in which the galloyl derivative of quercetin was identified for the first time in I. laurina fruit peels. GC–MS enabled separating and identifying substances such as palmitic and stearic acids, and ethyl oleate. It is possible to conclude that I. laurina pulp can be a supplementary food as a source of phenolic compounds, and the other organs of the plant (leaves and peel) are rich in flavonoids with great antioxidant capacity, making this species a promising source of antioxidants.

New anti-α-Glucosidase and Antioxidant Ingredients from Winery Byproducts: Contribution of Alkyl Gallates.

Wine-making activity entails the production of solid and semisolid byproducts (grape stems and pomace and wine lees) that negatively impact the environment and industrial sustainability. Their features as sources of bioactive compounds support valorization procedures for functional and healthy ingredients. This work uncovers the quantitative alkyl gallates (gallic acid esters, C1-C12) profile of fresh (freeze-dried) materials and the effect of oven-drying on their stability by UHPLC-ESI-QqQ-MS/MS. The functionality was established concerning DPPH• scavenging and antihyperglycemic power. Wine lees exerted the highest high-free concentration of galloyl derivatives, ethyl gallate being the most abundant ester (3472.62 ng/g dw, on average). About the impact of the stabilization process, although as a general trend, the thermal treatment reduced the concentration, the reduction dimensions depended on the compound/matrix, remaining in valuable concentrations. Concerning radical scavenging, ze-dried stems and pomace displayed the highest capacity (24.11 and 18.46 mg TE/g dw, respectively), being correlated with propyl gallate (r2 = 0.690), butyl gallate (r2 = 0.686), and octyl gallate (r2 = 0.514). These two matrices exerted α-glucosidase inhibitory activity (1.58 and 1.46 units/L) equivalent to that of acarbose (a recognized α-glucosidase inhibitor). The newly described bioactive phytochemicals in winery residues (galloyl esters) and their correlation with functional traits allow for envisioning valorization alternatives.

Mass Spectrometric Fingerprint Mapping Reveals Species-Specific Differences in Plant Polyphenols and Related Bioactivities.

Plant species show large variation in the composition and content of their tannins and other polyphenols. These large metabolites are not easy to measure accurately, but they are important factors for species bioactivity and chemotaxonomy. Here, we used an automated group-specific UHPLC-DAD-MS/MS tool to detect and quantify eight most common polyphenol groups in 31 chemically diverse plant species representing many types of growth forms and evolutionary ages. Ten replicate plants were used for each species and two polyphenol-related bioactivities, i.e., protein precipitation capacity and oxidative activity were measured in all samples as well. By the help of a novel 2D fingerprint mapping tool we were able to visualize the qualitative and quantitative differences between the species in hydrolysable tannins (galloyl and hexahydroxydiphenoyl derivatives), proanthocyanidins (procyanidins and prodelphinidins), flavonols (kaempferol, quercetin and myricetin derivatives) and quinic acid derivatives together with the two bioactivities. The highest oxidative activities were found with species containing ellagitannins (e.g., Quercus robur, Geranium sylvaticum, Lythrum salicaria and Chamaenerion angustifolium) or prodelphinidin-rich proanthocyanidins (e.g., Ribes alpinum, Salix phylicifolia and Lysimachia vulgaris). The best species with high protein precipitation capacity were rich in gallotannins (Acer platanoides and Paeonia lactiflora) or oligomeric ellagitannins (e.g., Comarum palustre, Lythrum salicaria and Chamaenerion angustifolium). These types of tools could prove their use in many types of screening experiments and might reveal even unusually active polyphenol types directly from the crude plant extracts.

Seasonal Variation in Plant Polyphenols and Related Bioactivities across Three Years in Ten Tree Species as Visualized by Mass Spectrometric Fingerprint Mapping.

The currently changing climates and environments place plants under many types of stresses that affect both their survival and levels of chemical defenses. The gradual induction of defenses in stressed plant populations could be monitored on a yearly basis unless a seasonal and yearly variation in natural defense levels obscures such monitoring schemes. Here, we studied the stability of the species-specific polyphenol composition and content of 10 tree species over three growing seasons using five replicate trees per species. We specifically measured hydrolyzable tannins (galloyl and hexahydroxydiphenoyl derivatives), proanthocyanidins (procyanidins and prodelphinidins), flavonols (kaempferol, quercetin and kaempferol derivatives) and quinic acid derivatives with the group-specific UHPLC-DAD-MS/MS tool, together with two bioactivities, the protein precipitation capacity and oxidative activity. With the help of a fingerprint mapping tool, we found out that species differed a lot in their seasonal and between-year variation in polyphenols and that the variation was also partially specific to compound groups. Especially ellagitannins tended to have declining seasonal patterns while the opposite was true for proanthocyanidins. Some of the species showed minimal variation in all measured variables, while others showed even induced levels of certain polyphenol groups during the 3-year study. For every species, we found either species-specific baseline levels in qualitative and quantitative polyphenol chemistry or the compound groups with the most plasticity in their production. The used tools could thus form a good combination for future studies attempting to monitor the overall changes in polyphenol chemistry due to various biotic or abiotic stress factors in plant populations or in more controlled environments.

1,5-Anhydro-d-glucitol derivative and galloylated flavonoids isolated from the leaves of Acer ginnala Maxim. as dual inhibitors of PTP1B and α-glucosidase enzymes: In vitro and in silico studies

Four undescribed compounds (two 1,5-anhydro-d-glucitol derivatives and two galloyl derivatives) and fourteen known compounds were isolated and structurally identified from leaves of Acer ginnala Maxim. (Amur maple). Structures and absolute configurations of the four undescribed compounds were determined using extensive analysis of NMR spectroscopic, HRESI-MS, modified Mosher ester method, and comparison with spectroscopic data of known compounds. Bioactivity evaluation revealed that the isolated 1,5-anhydro-d-glucitol derivative, galloylated flavonol rhamnosides, and galloylated flavanols had inhibitory effects on both protein tyrosine phosphatase-1B (PTP1B, IC50 values ranging of 3.46–12.65 μM) and α-glucosidase (IC50 values ranging of 0.88–6.06 μM) in comparison with a positive control for PTP1B (ursolic acid, IC50 = 5.10 μM) or α-glucosidase (acarbose, IC50 = 141.62 μM). A combination of enzyme kinetic analysis and molecular docking provided additional evidence in favor of their inhibitory activities and mechanism. These data demonstrate that A. ginnala Maxim. together with its constituents are promising sources of potent candidates for developing novel anti-diabetic medications.

Synthesis, Characterization, Antiproliferative Activity of Galloyl Derivatives and Investigation of Cytotoxic Properties in HepG2/C3A Cells.

Appropriate substituents in the galloyl group could lead to significant biological properties. Novel galloyl-substituted compounds bearing 2-substituted-1, 3, 4-oxadiazol-5-yl, 5- substituted-1,2,4-triazol-3-yl, and carboxamide groups were synthesized and evaluated for their antiproliferative activity. Additionally, galloyl hydrazide (2) was evaluated by performing cytotoxicity, membrane integrity, cell cycle, and apoptosis assays in HepG2/C3A cells. General procedure was used for the synthesis of galloyl-substituted (3-9, 11) and characterized by their spectroscopic data (1H and 13C NMR). The antiproliferative activity of all novel galloyl derivatives was evaluated against nine human tumors and one nontumoral cell line. Three response parameters (GI50, TGI, and LC50) were calculated. The cytotoxicity test was performed for the resazurin assay. The membrane integrity, cell cycle, and apoptosis assays were performed by flow cytometry. The substitution of the methoxy group of the galloyl ring system for a carboxamide group (3, 4, 5, and 6) produced compounds with moderate antitumoral activity, particularly 6, against six human cancer cell lines, K-562, PC-3, NCI-ADR/RES, OVCAR, 786-0 and NCI-H460, with GI50 values ≤ 9.45 μg/mL. Triazole derivatives 7 and 8 exhibited higher antitumoral activity toward OVCAR, MCF-7 and leukemia K-562 cell lines, exhibiting GI50 values less than 10 μg/mL. Compound 11 displayed significant activity against PC-3 (GI50 = 4.31 μg/mL), OVCAR (GI50 = 8.84 μg/mL) and K-562 (GI50 = 8.80 μg/mL) cell lines. Galloyl hydrazide (2) had cytotoxic activity in HepG2/C3A cells (IC50 = 153.7 μg/mL). In membrane permeability, cell count, cell cycle, and apoptosis assays, as determined using the IC50 of compound (2) in HepG2/C3A cells, increased membrane permeability, decreased cell count, altered cell cycle, and initial apoptosis was observed compared to the control group. Thus, our results showed for the first time the synthesis, antiproliferative activity, and cytotoxicity of galloyl-substituted compounds. Galloyl-substitution does not have a very strong synergistic effect in the inhibition of cancer cell proliferation compared with galloyl hydrazide (2). Compound 2 demonstrated promising activity in HepG2/C3A hepatocarcinoma cells.

Design, synthesis, and biological evaluation of novel double-winged galloyl derivatives as HIV-1 RNase H inhibitors

Human immunodeficiency virus (HIV) reverse transcriptase (RT)-associated ribonuclease H (RNase H) remains as the only enzyme encoded within the viral genome not clinically validated as an antiviral target. We have previously reported that the galloyl derivative II-25 had RNase H inhibitory activity in enzymatic assays but showed weak antiviral activity in phenotypic assays due its large polarity and poor membrane permeability. In this report, we report on a series of II-25 derivatives, obtained by addition of different hydrophobic moieties (“the wings”) at the C-2 and C-3 positions of the piperazine ring that showed improved RNase H inhibitory activity. Six compounds showed strong inhibitory activity and were found to be more potent than β-thujaplicinol in enzymatic assays. The most potent compound was IA-6 and exhibited the best inhibitory activity (IC50 = 0.067 ± 0.02 μM). IA-6 was around 11 and 30 times more potent than II-25 and β-thujaplicinol, respectively. Molecular modeling studies predict a strong hydrophobic interaction between the furylmethylaminyl group of IA-6 and the side chain of His539, explaining the potent HIV-1 RNase H inhibition. Unfortunately, none of the derivatives showed significant antiviral activity in cell culture. It is worth emphasizing that most of the obtained compounds show low cytotoxicity (CC50 > 20 μM), which confirms the significance of identifying galloyl derivatives as valuable leads for further optimization.

In Vitro Cell Culture of Rhus coriaria L.: A Standardized Phytocomplex Rich of Gallic Acid Derivatives with Antioxidant and Skin Repair Activity

This study focused on the biological evaluation and chemical characterization of a new ingredient obtained by in vitro cell culture of Rhus coriaria L. An in vitro plant cell culture method permits to cultivate plant in a short period of time and to obtain extract with a high safety profile for the consumer, free from heavy metals, pesticides, aflatoxins, bacterial or fungal contamination. Through the selection of specific cell culture media, it was possible to obtain a Rhus coriaria cell line with a high content of gallic acid derivatives. The Rhus coriaria L. phytocomplex (RC-P), containing 7.6% w/w of acid gallic derivatives, was obtained by drying of plant cell biomass after 14 days of growth in the final selected culture medium. UPLC-ESI-MS and UPLC-DAD analysis allowed to identify numerous gallic acid derivatives, such as galloyl hexose, trigalloyl hexose and high molecular weight galloyl derivatives, and to quantify their overall content. The antioxidant activity of the RC-P was tested by DPPH assay and the wound healing activity was evaluated using a scratch wound healing test on human keratinocytes and fibroblasts. This work showed that RC-P could be a new effective cosmetic ingredient with antioxidant and skin repair activity.

Human Metabolites of Hamaforton™ (Hamamelis virginiana L. Extract) Modulates Fibroblast Extracellular Matrix Components in Response to UV-A Irradiation.

Hamamelis virginiana L. a rich source of both condensed and hydrolyzable tannins, utilized to treat dermatological disorders. Since no experimental and clinical data is available for its use as oral formulation in skin related disorders, the purpose of this study was to investigate the effects of Hamaforton™ (Hamamelis virginiana extract) metabolites on gene dysregulation induced by ultraviolet A radiation in cultured human dermal fibroblasts. A combination of in vivo and ex vivo experimental designs has been exploited in order to take into account the polyphenol metabolic transformation that occurs in humans. 12 healthy volunteers received either a capsule of Hamaforton™ or a placebo in a randomized, blinded crossover trial. After Hamaforton™ ingestion, the kinetic of appearance of galloyl derivatives was measured in plasma. Then, in the ex vivo experiment, the serum isolated after supplementation was used as a source of Hamaforton™ metabolites to enrich the culture medium of dermal fibroblasts exposed to ultraviolet A radiation. Three different gallic acid metabolites (4-O-methyl gallic acid, 4-O-methyl gallic acid sulphate and trimethyl gallic acid glucuronide) were identified in volunteer plasma. While, ultraviolet A irradiation of dermal fibroblasts affected the expression of extracellular matrix genes, the presence of Hamaforton™ metabolites in the culture media did not affect the expression of most of those genes. However, the activation of the expression of 10 different genes involved in repair processes for the maintenance of skin integrity, suggest that the metabolites can play a role in damage recovery. To our knowledge, this is the first study that demonstrates the bioavailability of Hamaforton™ phenolic compounds, and the effects of its metabolites on cultured dermal fibroblast response to ultraviolet A irradiation.

LC-ESI-QTOF-MS/MS Analysis, Cytotoxic, Antiviral, Antioxidant, and Enzyme Inhibitory Properties of Four Extracts of Geranium pyrenaicum Burm. f.: A Good Gift from the Natural Treasure.

This study focused on the biological evaluation and chemical characterization of Geranium pyrenaicum Burm. f. Different solvent extracts (hexane, ethyl acetate, methanol, and water extracts) were prepared. The phytochemical profile, antioxidant, and enzyme inhibitory activity were investigated. Cytotoxicity was assessed using VERO, FaDu, HeLa and RKO cells. The antiviral activity was carried out against HSV-1 (Herpes simplex virus 1) propagated in VERO cell line. The aqueous extract, possessing high phenolic content (170.50 mg gallic acid equivalent/g extract), showed the highest reducing capacity (613.27 and 364.10 mg Trolox equivalent/g extract, for cupric reducing antioxidant capacity and ferric reducing antioxidant power, respectively), radical scavenging potential (469.82 mg Trolox equivalent/g extract, against 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)), metal chelating ability (52.39 mg ethylenediaminetetraacetic acid equivalent/g extract) and total antioxidant capacity (3.15 mmol Trolox equivalent/g extract). Liquid chromatography-electrospray ionization-quadrupole time-of-flight-mass spectrometry (LC-ESI-QTOF-MS/MS) alloved to tentatively identify a total of 56 compounds in the extracts, including ellagitannins, gallic acid and galloyl derivatives amongst others. The ethyl acetate extracts substantially depressed cholinesterase enzymes (4.49 and 12.26 mg galantamine equivalent/g extract against AChE and BChE, respectively) and α-amylase enzyme (1.04 mmol acarbose equivalent/g extract). On the other hand, the methanolic extract inhibited tyrosinase (121.42 mg kojic acid equivalent/g extract) and α-glucosidase (2.39 mmol acarbose equivalent/g extract) activities. The highest selectivity towards all cancer cell lines (SI 4.5–10.8) was observed with aqueous extract with the FaDu cells being the most sensitive (CC50 40.22 µg/mL). It can be concluded that the presence of certain bioactive antiviral molecules may be related to the high anti HSV-1 activity of the methanolic extract. This work has generated vital scientific data on this medicinal plant, which is a prospective candidate for the creation of innovative phyto-pharmaceuticals.

Formation of Dehydrohexahydroxydiphenoyl Esters by Oxidative Coupling of Galloyl Esters in an Aqueous Medium Involved in Ellagitannin Biosynthesis.

Hexahydroxydiphenoyl (HHDP) and dehydrohexahydroxydiphenoyl (DHHDP) groups are the major acyl components of ellagitannins, which are polyphenols whose biosynthesis have attracted considerable attention; however, the mechanisms of the production of HHDP and DHHDP in the ellagitannin biosynthesis have not been clarified. With the aim of elucidating such a mechanism, this study investigates the CuCl2 -mediated oxidation of simple galloyl derivatives in an aqueous medium. It is shown that the oxidation of methyl gallate affords a DHHDP-type dimer, whose reduction with Na2 S2 O4 yields an HHDP-type dimer. However, the oxidation of the HHDP-type product over CuCl2 does not afford the parent DHHDP ester. The oxidation of 1,4-butanediol digallate under the same conditions produces a DHHDP-type product via the intramolecular coupling of galloyl groups. These results strongly suggest that the DHHDP group is the initial product of the oxidative coupling of two galloyl groups in the ellagitannin biosynthesis, and subsequent reductive metabolism affords HHDP esters.

Vibrational Analysis and Molecular Docking Studies on some Ribonuclease-H HIV Inhibitors

Ribonuclease-H (RNase-H) enzyme of the human immunodeficiency virus (HIV) reverse transcriptase (RT) shows inhibitory activity with novel galloyl derivatives having enzymatic assays of IC50 S at sub to low micromolar concentration. The computational analysis of these stated galloyl derivatives was carried out in order to extract information and performance with the target proteins. The compounds N-hydroxylisoquinolinediones (HID), β-thujaplicinol, diketoacid, diketoacid ester, pyrimidinol carboxylic acids, naphthyridinones, 3hydroxypyrimidine-2,4-dione (HPD), and hydroxypyridonecarboxylic acids are the selected galloyl derivatives of human immunodeficiency virus-I (HIV-I) RNase-H active site inhibitors that were optimized using Turbomole software. Further evaluation of their NMR shielding of the stated compounds was performed using B3-LYP hybrid functional, and the def-SV basis set was carried out from the same software. These optimized compounds were then docked to targets (PDB Id: 5EGA and 3K2P) using AutoDock 4 software. After analyzing the docking result, Hydroxylisoquinoline and Naphthyridinones give better binding results with both the targets.

Chemosystematic implications based on metabolic profiling of the genus Byrsonima (Malpighiaceae)

The objective of this study was to evaluate the phytochemical profile of Byrsonima using metabolomics and high-resolution mass spectrometry to better understand the chemical diversity in this clade, comparing these results to the classification and more recent phylogenetic systematics of the genus. We analysed fragments of stem bark from 42 specimens of fifteen Byrsonima species using HPLC-DAD-MS. The species used were Byrsonima affinis, B. arthropoda, B. basiloba, B. chrysophylla, B. coccolobifolia, B. crassifolia, B. cydoniifolia, B. intermedia, B. pachyphylla, B. rigida, B. salzmanniana, B. sericea, B. spicata, B. umbellata and B. verbascifolia, derived from two traditional subgenera of Byrsonima: subg. Byrsonima and subg. Macrozeugma. The data were processed, aligned and evaluated statistically using multivariate analysis. Thirty-seven compounds were characterized, confirming the homogeneous chemical profile of Byrsonima and that the subgeneric classification should be reviewed. The chemical homogeneity in Byrsonima could be related to the presence of galloyl esters, which inhibit oxidation reactions for the biosynthesis of secondary metabolites. Galloyl ester compounds have been considered chemical markers of recently diversified groups and galloyl derivatives could be an important strategy for plant survival. The results also suggest that this genus belongs to a recent lineage within the Malpighiaceae.

Optimization of a Green Ultrasound-Assisted Extraction of Different Polyphenols from Pistacia lentiscus L. Leaves Using a Response Surface Methodology.

Pistacia lentiscus leaves are used in several applications, thanks to their polyphenolic abundance. Thiswork aimed to characterize the polyphenols and to optimize the extraction conditions to shorten the time, decrease the consumption of solvent, and to maximize the yield of different classes of phenolics, which have diverse industrial applications. The variables were optimized by applying a Box–Behnken design. Galloyl and myricetin derivatives were the most abundant compounds, and two new tetragalloyl derivatives were identified by LC-MS/MS. According to the models, the maximum yields of polyphenols (51.3 ± 1.8 mg g−1 DW) and tannins (40.2 ± 1.4 mg g−1 DW) were obtained using 0.12 L g−1 of 40% ethanol at 50 °C. The highest content of flavonoids (10.2 ± 0.8 mg g−1 DW) was obtained using 0.13 L g−1 of 50% ethanol at 50 °C, while 0.1 L g−1 of 30% ethanol at 30 °C resulted in higher amounts of myricitrin (2.6 ± 0.19 mg g−1 DW). Our optimized extraction decreased the ethanolic fraction by 25% and halved the time compared to other methods. These conditions can be applied differently to obtain P. lentiscus extracts richer in tannins or flavonoids, which might be employed for various purposes.

Acacia farnesiana pods (plant: Fabaceae) possesses anti-parasitic compounds against Haemonchus contortus in female lambs

Acacia farnesiana pods are rich in secondary metabolites and their biological activities have been recorded as antibacterial, antioxidant and anthelmintic. Previously, an in vitro bioguided study showed the important ovicidal and larvicidal effects of an organic fraction (EtOAc-F) from a hydroalcoholic extract of A. farnesiana pods against Haemonchus contortus. The present study aimed to assess the in vivo anthelmintic effect of EtOAc-F from A. farnesiana pods on the H. contortus faecal egg elimination in female lambs and on the infective larvae (L3) population reduction in coprocultures. The EtOAc-F was obtained from a hydroalcoholic extract from A. farnesiana pods through chromatographic procedures; additionally, some secondary compounds were identified using high-performance liquid chromatography (HPLC). Twenty-one ‘Katahdin’ crossbred female lambs ranging from three to four months of age, with body weights 21.9 ± 0.39 kg were used. Animals were orally infected with H. contortus (L3) by a single dose of 350 L3/kg BW. Three experimental groups (n = 7) were assigned as follows: 1) Control (untreated), 2) Albendazole, as a positive control (at 7.5 mg/kg BW, unique dose) and 3) EtOAc-F (at 100 mg/kg BW, once every third day, with three applications in total). Individual faecal samples were collected once a week for 5 weeks (at days 38, 45, 52, 59 and 66) post-treatment, to measure the faecal egg counts (FEC) and to obtain the H. contortus (L3) population from faecal cultures. The highest FEC reduction caused by EtOAc-F was 67.7%; meanwhile, albendazole showed a total FEC reduction after the second week post-treatment (day 45). On the other hand, the fraction caused an important reduction in the larval population in coprocultures (54.3–68.5%). The phytochemical analysis revealed the presence of galloyl derivatives and flavonoids as major compounds. The A. farnesiana pods could serve as a natural anthelmintic for the control of H. contortus, and perhaps for controlling other parasites of veterinary importance.

Phenolic profiling, biological activities and in silico studies of Acacia tortilis (Forssk.) Hayne ssp. raddiana extracts

To evaluate the phenolic profile and biological activity of the Algerian Sahara plant Acacia tortilis (Forssk.) Hayne ssp. raddiana decoction and 80% ethanolic extracts were studied. Chromatographic profiling indicated the presence of 36 phenolic compounds, including gallic acid esterified derivatives, galloylquinic derivatives and flavan-3-ols galloyl derivatives. Both extracts showed significant cytotoxic activity and a potent anti-inflammatory activity. They were effective against several multi-drug resistant bacteria, namely methicillin-resistant Staphylococcus aureus (MRSA). To understand the possible mechanism of action of MRSA inhibition activity, an in silico docking analysis was done using a virtual library of the 36 determined phenolic compounds against penicillin-binding protein (PBP2a), a protein known to be involved in MRSA resistance to β-lactam antibiotics. A. tortilis extracts showed interesting biological activities and the phenolic compounds found could be an interesting starting point for the development of cytotoxic and anti-inflammatory drugs and especially anti-MRSA antibiotics.

Metabolite profiling and antidiabetic attributes of ultrasonicated leaf extracts of Conocarpus lancifolius

Objective: To profile the secondary metabolites and to evaluate the antidiabetic potential of hydroethanolic leaf extracts of Conocarpus lancifolius.Methods: The various hydroethanolic extracts of Conocarpus lancifolius leaf were prepared by ultrasonication assisted freeze- drying. Total phenolic contents, flavonoid contents, antioxidant activity, α-glucosidase and α-amylase inhibitions of leaf extracts were determined. The metabolite profiling was accomplished by UHPLC-Q-TOF-MS/MS analysis. The antidiabetic assessment of the most potent extract was carried out by measuring the hypoglycemic and hypolipidemic effect in the high fat diet-fed diabetic albino mice. The blood glucose level, haemoglobin, total cholesterol, high-density lipoproteins (HDL) and low-density lipoproteins (LDL) were determined.Results: The 60% ethanolic extract exhibited the highest phenolic and flavonoid contents of (349.39 ± 2.13) mg GAE/g dry extract and (116.95 ± 2.34) mg RE/g dry extracts, respectively, and the highest DPPH scavenging activity with an IC50 value of (32.87 ± 1.11) μg/ mL. The IC50 values for α-glucosidase and α-amylase inhibitions were (38.64 ± 0.93) μg/mL and (44.80 ± 1.57) μg/mL, respectively. UHPLC-Q-TOF-MS/MS analysis confirmed the presence of gallic acid, ellagic acid, corilagin, kaempherol-3-O-rutinoside, caffeic acid derivative, isorhamnetin and galloyl derivatives in the 60% ethanolic extract. Plant extract at a dose of 450 mg/kg body weight reduced blood glucose level, total cholesterol, LDL and HDL, and increased haemoglobin in alloxan-induced diabetic mice,Conclusions: Conocarpus lancifolius leaves are proved as a good source of biologically functional metabolites and possess antidiabetic activity which may be further explored to treat diabetes.

Galloyl flavonoids from Acacia farnesiana pods possess potent anthelmintic activity against Haemonchus contortus eggs and infective larvae.

Acacia farnesiana (L.) Willd is a shrub legume used as condiment, medicinal plant and bioactive herbage. This species is used in traditional medicine of several countries to relieve the symptoms of gastrointestinal diseases, diarrhoea, stomach pain and typhoid as well as astringent, antidysenteric and anthelmintic. Some studies have shown that this plant displayed anthelmintic activity against several gastrointestinal nematode parasites of livestock, and also against parasites of human beings, such as malaria. This work describes the isolation and chemical identification of the anthelmintic compounds of Acacia farnesiana pods against eggs and infective larvae of the sheep parasitic nematode Haemonchus contortus. The bio-guided chemical fractioning of A. farnesiana pods using ethyl acetate against H. contortus eggs and infective larvae allowed for the identification of naringenin 7-O-(6″-galloylglucoside) (flavonol group) as the compound responsible for the anthelmintic activity against this important parasitic nematode. Anthelmintic activity was assessed using the egg hatching inhibition assay (EHI) and mortality tests. A complete hydroalcoholic extract (HA-E) at 12.5-50mg/mL, an aqueous fraction (Aq-F) at 3.12-25mg/mL and an ethyl acetate fraction (EtOAc-F) at 3.12-25mg/mL were analysed in the first selection phase. The purification of compounds through the chromatographic separation of the organic fraction resulted in nine less complex mixtures (C1F1, C1F2, C1F3, C1F4, C2F1, C2F2, C2F3, C2F4 and C2F5) that were assessed at 0.62-5mg/mL concentrations. In addition, thiabendazole (0.6mg/mL) and ivermectin (5 mg/mL) were used as positive controls. Likewise, distilled water and 4% methanol were used as negative controls. The bioactive compounds of EtOAc-F were obtained and characterised through chromatographic processes like open column chromatography, thin layer chromatography (TLC), high performance liquid chromatography (HPLC), ultra-performance liquid chromatography (UPLC) and gass chromatography-mass detection (GC-MS). Bioactive compounds were identified by spectroscopy (1H and 13C NMR) and mass spectrometric analysis. Additionally, the H. contortus eggs and infective larvae exposed to the bioactive compounds were observed through environmental scanning electron microscopy (ESEM) and confocal laser scanning microscopy (CLSM). Data were analysed based on a completely randomised design using ANOVA through a general linear model. The EtOAc-F fraction showed the highest ovicidal and larvicidal activities, at close to 100% at 3.12 and 6.25mg/mL, respectively. The treatments C1F2, C1F3 and C2F3 displayed the main ovicidal activity (80-100%) at 2.5mg/mL. The major compounds found in these sub-fractions were identified as galloyl derivatives and flavanones, including gallic acid (1), methyl gallate (2), ethyl gallate (3), naringin (4), naringenin 7-O-(4″, 6″-digalloylglucoside) (5), naringenin 7-O-(6″-galloylglucoside) (6) and naringenin (7). Likewise, the ESEM and CLSM images showed that the assessed compounds adhered to the eggshell and the external cuticle of the larvae. These results indicate that A. farnesiana pods contain nematocidal compounds and might be promising natural anthelmintic agents against H. contortus. This leguminous plant could be used as a nutraceutical food source for the control of gastrointestinal nematodes in small ruminants.