Denise Brentan Silva

Isolamento e avaliação da atividade citotóxica de alguns alcalóides oxaporfínicos obtidos de annonaceae

A different methodology was used to isolate and purify oxoaporphine alkaloids, as they are difficult to separate by the usual workup when in mixture. Alkaloid extracts from Annonaceae species were obtained by base/acid extraction. The extracts were concentrated and submitted to partition in solutions of acids of different pKa values, followed by separation by preparative TLC using 1 mm thick silica gel impregnated with oxalic acid (11.2% w/w). Liriodenine, lisycamine, lanuginosine, and O-methylmoschatoline were obtained and tested against tumoral cells (line Hep2, ATCC-CCL 23, larynx carcinoma). Only O-methylmoschatoline (IC50 12.4 µM) was more active than cisplatin (18.0 µM).

Variação química no óleo essencial das folhas de seis indivíduos de Duguetia furfuracea (Annonaceae)

Duguetia furfuracea, Annonaceae, a pasture weed, is also used in folk medicine in several Brazilian states. Because of the significant difference in odor between two groups of this plant in a remnant patch of savanna in Campo Grande county, Mato Grosso do Sul, six samples were selected according to the odor intensity in the leaves (In-01, In-03, and In-05: pronounced odor; In-02, In-04, and In-06: weak odor or its absence). The leaves were collected and subjected to steam distillation for extraction of essential oils, which were analyzed by GC-MS. A good agreement was found between CG/MS results and olfactory evaluation of the samples: specimens exhibiting leaf scent had a high percentage of monoterpenes and some sesquiterpenes, as follows: In-01 (β-phellandrene, 42.2%; myrcene, 6.8%; α-phellandrene, 4.6%); In-03 (terpin-4-ol, 21.6%; sabinene, 17.3%; p-cymene, 5.6%); In-05 (sabinene, 25.1%; terpin-4-ol, 16.2%; p-cymene, 8.3%). Only sesquiterpenes were found in the specimens having weakly scented or scentless leaves (main constituent bicyclogermacrene: 21.4%, 24.0%, and 29.1%, respectively, for In-02, In-04 and In-06).

Leaf and Root Extracts from Campomanesia adamantium (Myrtaceae) Promote Apoptotic Death of Leukemic Cells via Activation of Intracellular Calcium and Caspase-3

Phytochemical studies are seeking new alternatives to prevent or treat cancer, including different types of leukemias. Campomanesia adamantium, commonly known as guavira or guabiroba, exhibits pharmacological properties including antioxidant, antimicrobial, and antiproliferative activities. Considering the anticancer potential of this plant species, the aim of this study was to evaluate the antileukemic activity and the chemical composition of aqueous extracts from the leaves (AECL) and roots (AECR) of C. adamantium and their possible mechanisms of action. The extracts were analyzed by LC-DAD-MS, and their constituents were identified based on the UV, MS, and MS/MS data. The AECL and AECR showed different chemical compositions, which were identified as main compounds glycosylated flavonols from AECL and ellagic acid and their derivatives from AECR. The cytotoxicity promoted by these extracts were evaluated using human peripheral blood mononuclear cells and Jurkat leukemic cell line. The cell death profile was evaluated using annexin-V-FITC and propidium iodide labeling. Changes in the mitochondrial membrane potential, the activity of caspases, and intracellular calcium levels were assessed. The cell cycle profile was evaluated using propidium iodide. Both extracts caused concentration-dependent cytotoxicity only in Jurkat cells via late apoptosis. This activity was associated with loss of the mitochondrial membrane potential, activation of caspases-9 and -3, changes in intracellular calcium levels, and cell cycle arrest in S-phase. Therefore, the antileukemic activity of the AECL and AECR is mediated by mitochondrial dysfunction and intracellular messengers, which activate the intrinsic apoptotic pathway. Hence, aqueous extracts of the leaves and roots of C. adamantium show therapeutic potential for use in the prevention and treatment of diseases associated the proliferation of tumor cell.

Caatinga plants: Natural and semi-synthetic compounds potentially active against Trichomonas vaginalis.

Trichomonas vaginalis causes trichomoniasis; the most common but overlooked non-viral sexually transmitted disease worldwide. The treatment is based at 5-nitroimidazoles, however, failure are related to resistance of T. vaginalis to chemotherapy. Caatinga is a uniquely Brazilian region representing a biome with type desert vegetation and plants present diverse biological activity, however, with few studies. The aim of this study was to investigate the activity against T. vaginalis of different plants from Caatinga and identify the compounds responsible by the activity. A bioguided fractionation of Manilkara rufula was performed and four major compounds were identified: caproate of α-amyrin (1b), acetate of β-amyrin (2a), caproate of β-amyrin (2b), and acetate of lupeol (3a). In addition, six derivatives of α-amyrin (1), β-amyrin (2) and lupeol (3) were synthesized and tested against the parasite. Ursolic acid (5) reduced about 98% of parasite viability after 2h of incubation and drastic ultrastructural alterations were observed by scanning electron microscopy. Moreover, 5 presented high cytotoxicity to HMVII and HeLa cell line and low cytotoxicity against Vero line at 50 μM (MIC against the parasite). Metronidazole effect against T. vaginalis resistant isolate was improved when in association with 5.

Metabolomics as a tool for understanding the evolution of Tabebuia sensu lato

BackgroundPlant systematic studies have changed substantially in the last years, stimulated by new strategies for phylogenetic studies. In this regard, chemistry data has been a useful tool for understanding plant phylogenetic relationships.ObjectiveOur aim was to apply metabolomic approaches, followed by multivariate statistical analysis and dereplication of Tabebuia sensu lato species, and compare our results with classifications based on traditional taxonomy and molecular phylogeny. We also evaluated the application of metabolomics as a chemotaxonomic identification tool, as well as to enlighten plant chemical evolution.MethodsMetabolomic data was generated through a high-resolution mass spectrometry with electrospray ionization of 27xa0Tabebuia sensu lato specimens from different populations, consisting of 15 Handroanthus (from four species) and 12 Tabebuiasensu stricto (from three species). Chemometric tools, such as principal component analysis and metabolite heatmaps, were used to scrutinize the metabolic changes among species.ResultsTabebuia and Handroanthus species presented different secondary metabolite storage capacity. The genus Tabebuia revealed higher levels of glycosylated iridoids esterified with a phenylpropanoid moiety, such as specioside, verminoside, and minecoside, while Handroanthus accumulated iridoids linked to a simple phenol, lignans, and verbascoside derivatives.ConclusionThese results corroborate splitting the Tabebuia s.l., which was supported by profound changes in secondary metabolism, suggesting metabolomics as an excellent tool for understanding species evolution.

New cascarosides from Rhamnus purshiana and fragmentation studies of the class by IT-MS

RATIONALEnAnthrone and oxanthrone are important anthraquinone derivatives present in medicinal plants which are used in therapeutics as laxatives. Some of these plants need to be stored at least one year before they can be used in order to oxidize anthrones into oxanthrones, so to avoid severe diarrhea and dehydration. Therefore, this work aimed to characterize fragmentation reactions between these anthraquinones to provide an easy way to differentiate between the two classes, since it is necessary and important to discriminate and identify these derivatives in laxative plants and phytotherapic drugs.nnnMETHODSnAnthrone (cascarosides A-D) and oxanthrone (10-hydroxycascaroside A and B) derivatives were isolated and identified by NMR (1 H, 13 C, DEPT, NOESY) and used for fragmentation study by direct infusion on an electrospray ionization (ESI) ion trap mass spectrometer (AmazonSL, Bruker) in positive and negative mode.nnnRESULTSnThe additional hydroxyl at C-10 in oxanthrones allowed McLafferty-type rearrangements to form the quinone group in positive mode, while in negative mode the second sugar loss infringed the odd-electron rule and formed a radical fragment. No differences in fragmentation reactions were found between diastereoisomeric pairs, although the additional oxygen at C-10 of oxanthrones allowed a different fragmentation pattern.nnnCONCLUSIONSnThe proposed fragmentation patterns can be used to differentiate anthrones from oxanthrones in both ion modes. In addition, they can be applied to differentiate these compounds in anthraquinone-rich plants and phytotherapic drugs. Finally, herein, the strategy applied allowed us to identify new natural products. Copyright

Antiviral Activity of Crude Hydroethanolic Extract from Schinus terebinthifolia against Herpes simplex Virus Type 1

Herpes simplex virus infections persist throughout the lifetime of the host and affect more than 80u200a% of the humans worldwide. The intensive use of available therapeutic drugs has led to undesirable effects, such as drug-resistant strains, prompting the search for new antiherpetic agents. Although diverse bioactivities have been identified in Schinus terebinthifolia, its antiviral activity has not attracted much attention. The present study evaluated the antiherpetic effects of a crude hydroethanolic extract from the stem bark of S. terebinthifolia against Herpes simplex virus type 1 in vitro and in vivo as well as its genotoxicity in bone marrow in mammals and established the chemical composition of the crude hydroethanolic extract based on liquid chromatography-diode array detector-mass spectrometry and MS/MS. The crude hydroethanolic extract inhibited all of the tested Herpes simplex virus type 1 strains in vitro and was effective in the attachment and penetration stages, and showed virucidal activity, which was confirmed by transmission electron microscopy. The micronucleus test showed that the crude hydroethanolic extract had no genotoxic effect at the concentrations tested. The crude hydroethanolic extract afforded protection against lesions that were caused by Herpes simplex virus type 1 in vivo. Liquid chromatography-diode array detector-mass spectrometry and MS/MS identified 25 substances, which are condensed tannins mainly produced by a B-type linkage and prodelphinidin and procyanidin units.

Antibacterial activity of Limonium brasiliense (Baicuru) against multidrug-resistant bacteria using a statistical mixture design

ETHNOPHARMACOLOGICAL RELEVANCEnLimonium brasiliense (Boiss.) Kuntze (Plumbaginaceae) is commonly known as baicuru or guaicuru and preparations of its dried rhizomes have been popularly used in the treatment of premenstrual syndrome and menstrual disorder, and as an antiseptic in genito-urinary infections. This study evaluated the potential antibacterial activity of rhizome extracts against multidrug-resistant bacterial strains using statistical mixture design.nnnMATERIALS AND METHODSnThe statistical design of four components (water, methanol, acetone, and ethanol) produced 15 different extracts and also a confirmatory experiment, which was performed using water:acetone (3:7, v/v). The crude extracts and their ethyl-acetate fractions were tested against vancomycin-resistant Enterococcus faecium (VREfm), methicillin-resistant Staphylococcus aureus (MRSA) and Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae, all of which have been implicated in hospital and community-acquired infections. The dry residue, total polyphenol, gallocatechin and epigallocatechin contents of the extracts were also tested and statistical analysis was applied in order to define the fit models to predict the result of each parameter for any mixture of components. The principal component and hierarchical clustering analyses (PCA and HCA) of chromatographic data, as well as mass spectrometry (MS) analysis were performanced to determine the main compounds present in the extracts.nnnRESULTSnThe Gram-positive bacteria were susceptible to inhibition of bacterial growth, in special the ethyl-acetate fraction of ternary extracts from water:acetone:ethanol and methanol:acetone:ethanol against, respectively, VREfm (MIC=19µg/mL) and MRSA (MIC=39µg/mL). On the other hand, moderate activity of the ethyl-acetate fractions from primary (except water), secondary and ternary extracts (MIC=625µg/mL) was noted against KPC. The quadratic and special cubic models were significant for polyphenols and gallocatechin contents, respectively. Fit models to dry residue and epigallocatechin contents were not possible. PCA and HCA of the chromatographic fingerprints were disturbed by displacement retention time of some peaks, but the ultraviolet spectra indicated the homogeneous presence of flavan-3-ols characteristic of tannins. The MS confirmed the presence of gallic acid, gallocatechin, and epigallocatechin in extracts, and suggested the presence of monomers and dimers of B- and A-type prodelphinidins gallate, as well as a methyl gallate.nnnCONCLUSIONnOur results showed the antibacterial potential of L. brasiliense extracts against multidrug-resistant Gram-positive bacteria, such as VREfm and MRSA. The statistical design was a important tool to evaluate the biological activity by optimized form. The presence of some phenolic compounds was also demonstrated in extracts.

The Caatinga endemic Manilkara rufula possesses remarkable activity against Trichomonas vaginalis and Tritrichomonas foetus.

Tritrichomonas foetus infects the bovine urogenital tract, causing bovine trichomoniasis. Significant economic losses may occur due to infertility and abortion among cattle. Trichomonas vaginalis is the causative agent of trichomoniasis; the most common but overlooked non-viral sexually transmitted disease worldwide. Human and bovine trichomoniasis present treatment restrictions and efforts to identify new alternatives are essential. The present study evaluated the anti-trichomonads activities of seven fractions from northwest endemic plant Manilkara rufula. Flavonoids and condensed tannins were identified from these fractions by LC-DAD-MS/MS and MALDI-MS/MS. Altogether, the results demonstrated for the first time the structural description of tannins from leaves of M.xa0rufula and the relation of these compounds with anti-T.xa0vaginalis and anti-T.xa0foetus activities. Overall, this report reveals the potential of M.xa0rufula fractions against both parasites and shows new alternatives to treat the infection caused by trichomonads.

Simultaneous Determination of Enrofloxacin, Silver Sulfadiazine, Hydrocortisone Acetate, Hydrocortisone Sodium Succinate, and Preservative Excipients in Pharmaceutical Preparations Using HPLC–DAD Method

An HPLC–DAD method for simultaneous determination of enrofloxacin (ENR), silver sulfadiazine (SSD), hydrocortisone acetate (HAC), hydrocortisone sodium succinate (HSS), and preservative excipients common in liquid pharmaceutical forms was developed and validated. Dionex® chromatograph and C18 column were used at room temperature. The mobile phase was acetonitrile and water mixture (48:52 v/v) at pH 3.0. Isocratic elution mode was used at flow rate of 1.0xa0mLxa0min−1 and wavelength detections were performed at 240, 255, 270, and 280xa0nm. The method was validated by following international guidelines for evaluating selectivity, linearity, accuracy, precision, robustness, and limits of quantification and detection. Symmetrical chromatograph peaks were obtained in less than 6xa0min. All four calibration curves were linear. Relative standard deviation of drug contents was less than 2.0% with respect to intermediate precision and repeatability. Mean recoveries with respect to accuracy were between 98 and 102% for all four drugs. The limits of detection and quantification were, respectively, 0.024 and 0.072xa0μgxa0mL−1 for ENR, 0.035 and 0.105xa0μgxa0mL−1 for SSD, 0.014 and 0.042xa0μgxa0mL−1 for HAC, and 0.016 and 0.047xa0μgxa0mL−1 for HSS. Robustness of method was evaluated by Plackett–Burman’s test, showing the method was significantly affected for only one of the 14 parameter variations. Using mass spectrometry, HSS hydrolysis degradation product (i.e., free hydrocortisone) was identified both in standard chemical reference and in commercial formulation. In addition, the proposed method also was able to separate all drug chromatography peaks of one degradation product and two preservative excipients.Graphical Abstract