Gouregine, an α-Gem-Dimethyltetradehydrocularine Alkaloid, and Other Aporphinoid Alkaloids from the Bark of Guatteria olivacea (Annonaceae) and Their In Vitro Cytotoxic Activities.

Guatteria olivacea R.E. Fries is an Amazonian species known as 'envira-bobó' and 'envira-fofa' and is common in the states of Amazonas, Acre, and Pará. Recently, the essential oil from the leaves of this species has shown promising antitumor activity both in vitro and in vivo. The presence of isoquinoline-derived alkaloids, including aporphinoids and tetrahydroprotoberberine alkaloids, has also been previously reported. In our ongoing search for bioactive compounds from Annonaceae Amazonian plants, the bark of G. olivacea was investigated via classical chromatography techniques, which revealed nine compounds, eight isoquinoline-derived alkaloids, a rare alkaloid with a α-gem-dimethyltetradehydrocularine structure known as gouregine, seven known aporphinoid alkaloids: isopiline, O-methylisopiline, melosmine, 9-hydroxyiguattescine, dihydromelosmine, lysicamine, and guattouregidine, and one known pimaradiene diterpene: acanthoic acid. All the isolated compounds were described for the first time in the bark of G. olivacea, and their structures were elucidated by extensive analyses of their 1D and 2D NMR spectra in combination with MS data. The NMR data of the alkaloids isopiline, O-methylisopiline, melosmine, dihydromelosmine, and guattouregidine were revised due to incomplete data in the literature and some ambiguities. The in vitro cytotoxic activities of the isolated compounds were evaluated against human cancer (HepG2, KG-1a, and HCT116) and noncancerous (MRC-5) cell lines via the Alamar blue assay after 72 h of incubation. Among the compounds evaluated against human cancer cell lines, the most active was the oxoaporphine alkaloid lysicamine, which has strong activity against HCT116 cells, with an IC50 value of 6.64 µg/mL (22.79 µmol/L). Melosmine had a moderate effect on HCT116 cells, with an IC50 value of 16.77 µg/mL (49.70 µmol/L), whereas acanthoic acid had moderate effects on HepG2 and HCT116 cells, with IC50 values of 14.63 µg/mL (48.37 µmol/L) and 21.25 µg/mL (70.25 µmol/L), respectively.

Cupuaçu (Theobroma grandiflorum): A Multifunctional Amazonian Fruit with Extensive Benefits.

The Amazon region is known for its continental dimension, water abundance, and especially for the rich biodiversity that this biome hosts. Among the thousands of plant species in the Amazon, many represent food sources. Among these, cupuaçu (Theobroma grandiflorum (Willd. ex Spreng.) K.Schum.) stands out as an iconic fruit with an exotic flavor, appreciated for its remarkable organoleptic properties. The present review aims to provide a comprehensive description of its biology, agronomical uses, nutritional values, chemical compositions, medicinal properties, and industrial applications. The search based on scientific articles demonstrates T. grandiflorum as a valuable ingredient for the food, cosmetic and pharmaceutical sectors. Data analysis demonstrates that cupuaçu cultivation and processing contribute to the strengthening of local production chains and promotes the development of small communities, and thus the bioeconomy in the Amazon region. In this sense, since the last decade, cultivar improvement has required multidisciplinary efforts, resulting in disease-resistant plants with better productivity. Regarding its chemical composition, T. grandiflorum is a notable source of methylxanthine alkaloids, polyphenols, aroma compounds, and lipids. The presence of these compounds supports the use of cupuaçu in various products and help us to understand the potential health benefits of its consumption. Through the integration of all collected information, key gaps in basic and applied sciences were observed, highlighting the need for more research to uncover novel applications and products of T. grandiflorum. The development of new products based on biodiversity is fundamental to promoting environmental and economic sustainability, which are key steps to the survival of the Amazon rainforest. Therefore, this work summarizes the knowledge on this source and sheds light on a food source that is little known outside of the Amazon borders.

plantMASST - Community-driven chemotaxonomic digitization of plants.

Understanding the distribution of hundreds of thousands of plant metabolites across the plant kingdom presents a challenge. To address this, we curated publicly available LC-MS/MS data from 19,075 plant extracts and developed the plantMASST reference database encompassing 246 botanical families, 1,469 genera, and 2,793 species. This taxonomically focused database facilitates the exploration of plant-derived molecules using tandem mass spectrometry (MS/MS) spectra. This tool will aid in drug discovery, biosynthesis, (chemo)taxonomy, and the evolutionary ecology of herbivore interactions.

Pleonotoquinones, Cytotoxic Oxepinenaphthoquinones from Pleonotoma jasminifolia.

Two unusual naphthoquinones, named here as pleonotoquinones A (1) and B (2), were isolated along with two known anthraquinones (3 and 4) via chromatographic separations of an ethyl acetate extract of the roots of Pleonotoma jasminifolia. Compounds 1 and 2 are the first examples of quinones bearing a 2-methyloxepine moiety. The compounds were isolated with the aid of mass spectrometry and molecular networking, and their structures were resolved using 1D and 2D NMR and HRESIMS data. The isolated compounds were evaluated for their antiproliferative activity against human cancer cell lines, and compounds 1 and 2 displayed cytotoxicity against human colon cancer HCT116 cells (IC50 = 2.6 µM for compound 1 and IC50 = 4.3 µM for compound 2) and human liver cancer HepG2 cells (IC50 = 1.9 µM for compound 1 and IC50 = 6.4 µM for compound 2).

Essential oil and fenchone extracted from Tetradenia riparia (Hochstetter.) Codd (Lamiaceae) induce oxidative stress in Culex quinquefasciatus larvae (Diptera: Culicidae) without causing lethal effects on non-target animals.

We investigated the larvicidal activity of the essential oil (EO) from Tetradenia riparia and its majority compound fenchone for controlling Culex quinquefasciatus larvae, focusing on reactive oxygen and nitrogen species (RONS), catalase (CAT), glutathione S-transferase (GST), acetylcholinesterase (AChE) activities, and total thiol content as oxidative stress indicators. Moreover, the lethal effect of EO and fenchone was evaluated against Anisops bouvieri, Diplonychus indicus, Danio rerio, and Paracheirodon axelrodi. The EO and fenchone (5 to 25 µg/mL) showed larvicidal activity (LC50 from 16.05 to 18.94 µg/mL), followed by an overproduction of RONS, and changes in the activity of CAT, GST, AChE, and total thiol content. The Kaplan-Meier followed by Log-rank (Mantel-Cox) analyses showed a 100% survival rate for A. bouvieri, D. indicus, D. rerio, and P. axelrodi when exposed to EO and fenchone (262.6 and 302.60 µg/mL), while α-cypermethrin (0.25 µg/mL) was extremely toxic to these non-target animals, causing 100% of death. These findings emphasize that the EO from T. riparia and fenchone serve as suitable larvicides for controlling C. quinquefasciatus larvae, without imposing lethal effects on the non-target animals investigated.

Anti-biofilm and anti-inflammatory active diterpene isolated from the fruit of Xylopia benthamii R.E.Fr.

Xylopia benthamii (Annonaceae) is a plant with limited phytochemical and pharmacological evidence. Thus, using LC-MS/MS, we performed exploratory analyses of the fruit extract of X. benthamii, resulting in the tentative identification of alkaloids (1-7) and diterpenes (8-13). Through the application of chromatography techniques with the extract of X. benthamii, two kaurane diterpenes were isolated, xylopinic acid (9) and ent-15-oxo-kaur-16-en-19-oic acid (11). Their structures were established using spectroscopy (NMR 1D/2D) and mass spectrometry. The isolated compounds were submitted to anti-biofilm analysis against Acinetobacter baumannii, anti-neuroinflammatory and cytotoxic activity in BV-2 cells. Compound 11 (201.75 µM) inhibited 35% of bacterial biofilm formation and high anti-inflammatory activity in BV-2 (IC50 = 0.78 µM). In conclusion, the results demonstrated that compound 11 was characterized for the first time with pharmacological potential in the development of new alternatives for studies with neuroinflammatory diseases.

Anti-inflammatory and antioxidant properties of Bertholletia excelsa (H. B. K) bark extract / Propiedades antiinflamatorias y antioxidantes del extracto de corteza de Bertholletia excelsa (H. B. K)

Bertholletia excelsa is native to the Amazon Rainforest and is popularly known as the Brazil nut. It has socioeconomic importance due its nuts being a great export product. There are few studies in the literature regarding the biotechnological potential of its bark, although it is used in folk medicine. The aim of this study was to determine the chemical constituents, anti-inflammatory and antioxidant properties of B. excelsa bark extract (BEB). Twelve substances were identified by LC/MS/MS, and cytotoxicity tests were carried out, as well as analyses of nitric oxide production and elimination of free radicals. BEB caused cytoprotection against oxidative stress in macrophages, increased HMOX-1 expression, overcame the antioxidant effects of GPx-1 and reduced its expression and was able to inhibit leukocyte migration in use peritonitis. BEB efficiently attenuated oxidative stress due to its antioxidant and anti-inflammatory properties and, as such, can be used as a safe and effective source of a natural herbal medicine.

Bertholletia excelsa es originaria de la selva amazónica y se la conoce popularmente como nuez de Brasil. Tiene importancia socioeconómica debido a que sus frutos secos son un gran producto de exportación. Existen pocos estudios en la literatura sobre el potencial biotecnológico de su corteza, aunque se utiliza en medicina popular. El objetivo de este estudio fue determinar los componentes químicos, las propiedades antiinflamatorias y antioxidantes del extracto de corteza de B. excelsa (BEB). Se identificaron 12 sustancias mediante LC/MS/MS y se realizaron pruebas de citotoxicidad, así como análisis de producción de óxido nítrico y eliminación de radicales libres. BEB causó citoprotección contra el estrés oxidativo en macrófagos, aumentó la expresión de HMOX-1, superó los efectos antioxidantes de GPx-1 y redujo su expresión y fue capaz de inhibir la migración de leucocitos en la peritonitis de uso. BEB atenuó eficazmente el estrés oxidativo debido a sus propiedades antioxidantes y antiinflamatorias y, como tal, puede utilizarse como una fuente segura y eficaz de un medicamento a base de hierbas naturales.

Chemistry and Pharmacology of Bergenin or Its Derivatives: A Promising Molecule.

Bergenin is a glycosidic derivative of trihydroxybenzoic acid that was discovered in 1880 by Garreau and Machelart from the rhizomes of the medicinal plant Bergenia crassifolia (currently: Saxifraga crassifolia-Saxifragaceae), though was later isolated from several other plant sources. Since its first report, it has aroused interest because it has several pharmacological activities, mainly antioxidant and anti-inflammatory. In addition to this, bergenin has shown potential antimalarial, antileishmanial, trypanocidal, antiviral, antibacterial, antifungal, antinociceptive, antiarthritic, antiulcerogenic, antidiabetic/antiobesity, antiarrhythmic, anticancer, hepatoprotective, neuroprotective and cardioprotective activities. Thus, this review aimed to describe the sources of isolation of bergenin and its in vitro and in vivo biological and pharmacological activities. Bergenin is distributed in many plant species (at least 112 species belonging to 34 families). Both its derivatives (natural and semisynthetic) and extracts with phytochemical proof of its highest concentration are well studied, and none of the studies showed cytotoxicity for healthy cells.

Discovery of dual PKS involved in sclerotiorin biosynthesis in Penicillium meliponae using genome mining and gene knockout.

Fungi of the genus Penicillium section Sclerotiora have as their main characteristic the presence of orange-pigmented mycelium, which is associated with sclerotiorin, a chlorinated secondary metabolite of the azaphilone subclass of polyketides. Sclerotiorin presents anti-diabetes, antioxidant, anti-inflammatory, anti-Alzheimer, antiviral, and antimicrobial activities, which has always attracted the attention of researchers worldwide. During our ongoing search for azaphilone-producing Amazonian fungi, the strain of Penicillium MMSRG-058 was isolated as an endophyte from the roots of Duguetia stelechantha and showed great capacity for producing sclerotiorin-like metabolites. Using multilocus phylogeny, this strain was identified as Penicillium meliponae. Moreover, based on the genome mining of this strain through the reverse approach, a cluster of putative biosynthetic genes (BGC) responsible for the biosynthesis of sclerotiorin-like metabolites (scl cluster) was identified. The knockout of the sclA (highly reducing PKS) and sclI (non-reducing PKS) genes resulted in mutants with loss of mycelial pigmentation and terminated the biosynthesis of sclerotiorin-like metabolites: geumsanol B, chlorogeumsanol B, 7-deacetylisochromophilone VI, isochromophilone VI, ochrephilone, isorotiorin, and sclerotiorin. Based on these results, a biosynthetic pathway was proposed considering the homology of BGC scl genes with the azaphilone BGCs that have already been functionally characterized.

Toxicity of the essential oil from Tetradenia riparia (Hochstetter.) Codd (Lamiaceae) and its principal constituent against malaria and dengue vectors and non-target animals.

Malaria and dengue are diseases transmitted by mosquitoes of the genera Anopheles and Aedes resistant to commercial insecticides, which are toxic to non-target animals. Alternatively, eco-friendly strategies have focused on searching for essential oil (EO) from plants to control these mosquitoes. In this aspect, this study was carried out to investigate the toxicity of the EO from Tetradenia riparia and its main constituent against Anopheles and Aedes larvae and non-target animals Toxorhynchites haemorrhoidalis and Gambusia affinis. The mechanism of the larvicidal action of the EO and its main compound was investigated by the acetylcholinesterase (AChE) inhibition. The EO from T. riparia was extracted by hydrodistillation with yield of 1.4 ± 0.17%. The analysis of the EO by GC-MS and GC-FID revealed fenchone (38.62%) as the main compound. The EO (100 ppm) showed larvicidal activity against Anopheles and Aedes larvae (91 to 100% of mortality) (LC50 from 29.31 to 40.76 ppm). On the other hand, fenchone (10 ppm) showed more activity (89 to 100% of mortality) (LC50 from 5.93 to 7.00 ppm) than the EO. The EO and fenchone caused the inhibition of AChE (IC50 from 1.93 to 2.65 ppm), suggesting the inhibition of this enzyme as a possible mechanism of larvicidal action. Regarding toxicity, the EO (1000 ppm) and fenchone (100 ppm) showed low toxicity against T. haemorrhoidalis and G. affinis (9 to 74% of mortality) (LC50 from 170.50 to 924.89 ppm) (SI/PSF from 17.99 to 31.91) than the α-cypermethrin (0.52 ppm) which was extremally toxic against these non-target animals (100% of mortality, LC50 from 0.22 to 0.29 ppm). This significant larvicidal activity of the T. riparia EO and its main constituent, along with the low toxicity towards non-target organisms indicate these samples as a possible eco-friendly alternative for the control of malaria and dengue vectors.

Duguetia pycnastera Sandwith (Annonaceae) Leaf Essential Oil Inhibits HepG2 Cell Growth In Vitro and In Vivo.

Duguetia pycnastera Sandwith (Annonaceae) is a tropical tree that can be found in the Guyanas, Bolivia, Venezuela, and Brazil. In Brazil, it is popularly known as "ata", "envira", "envira-preta", and "envira-surucucu". In the present work, we investigated the in vitro and in vivo HepG2 cell growth inhibition capacity of D. pycnastera leaf essential oil (EO). The chemical composition of the EO was determined by GC−MS and GC−FID analyses. The alamar blue assay was used to examine the in vitro cytotoxicity of EO in cancer cell lines and non-cancerous cells. In EO-treated HepG2 cells, DNA fragmentation was measured by flow cytometry. The in vivo antitumor activity of the EO was assessed in C.B-17 SCID mice xenografted with HepG2 cells treated with the EO at a dosage of 40 mg/kg. Chemical composition analysis displayed the sesquiterpenes α-gurjunene (26.83%), bicyclogermacrene (24.90%), germacrene D (15.35%), and spathulenol (12.97%) as the main EO constituents. The EO exhibited cytotoxicity, with IC50 values ranging from 3.28 to 39.39 µg/mL in the cancer cell lines SCC4 and CAL27, respectively. The cytotoxic activity of the EO in non-cancerous cells revealed IC50 values of 16.57, 21.28, and >50 µg/mL for MRC-5, PBMC, and BJ cells, respectively. An increase of the fragmented DNA content was observed in EO-treated HepG2 cells. In vivo, EO displayed tumor mass inhibition activity by 47.76%. These findings imply that D. pycnastera leaf EO may have anti-liver cancer properties.

Amazonian Melastomataceae blueberries: Determination of phenolic content, nutritional composition, and antioxidant and anti-glycation activities.

Berries come from hundreds of different species of plants spread around the world. Blackberries, blueberries and raspberries, for instance, are popular berries that have attracted attention for providing several benefits to human health. Wild berries from the Melastomataceae family are commonly encountered in the Amazon, although these small blue fruits are poorly consumed. Although domesticated fruits give better monetary profits, the consumption of wild fruits is a desirable option to afford income and/or food to communities at the same time as keep the Amazon region preserved. Aiming the divulgation of the nutritional potential of these plants, this paper describes the study of six species of Amazonian blueberries, five of them from the Clidemia genus and one from the Tococa genus, in regard to their nutritional and chemical composition and antioxidant activity (AA). The levels of moisture, ash, protein, lipids, carbohydrates, and the total caloric values obtained for the Amazonian blueberries were comparable to other common edible berries. Although the six species are similar in terms of nutritional composition, their anthocyanin profiles and contents are quite peculiar. Two non-methylated anthocyanins, cyanidin and delphinidin, which bound to a variable number of sugars, characterized the berries of the genera Clidemia and Tococa. Clidemia japurensis, Clidemia hirta and Tococa bullifera were rich in tri-glycosylated anthocyanins, although differences are notable between them. Clidemia pustulata and Clidemia capitellata were characterized by the prevalence of mono-glycosylated anthocyanins, and Clidemia rubra showed a unique profile with mono- and di-glycosylated homologous as the main anthocyanins. In addition to their different chemical profiles, the concentrations of anthocyanins and other phenolic compounds varied a lot among the six species studied. The species C. rubra had the highest total concentration of phenolic acids and flavonoids. Therefore, this study showed that the blueberries analyzed have potential to be better explored, which we suggest doing in a sustainable way, aiming at the preservation of the Amazon's biodiversity.

Antitumor Effect of Guatteria olivacea R. E. Fr. (Annonaceae) Leaf Essential Oil in Liver Cancer.

Guatteria olivacea R. E. Fries (synonym Guatteria punctata (Aubl.) R.A. Howard) is a tree of 10-27 m tall popularly known as "envira-bobó", "envira-fofa", "envireira", "embira", "embira-branca", "embira-preta", envira-branca", and "envira-preta", which can be found in the Brazilian Amazon biome. In this study, we evaluated the cytotoxic and antitumor effects of the essential oil (EO) obtained from the leaves of G. olivacea against liver cancer using HepG2 cells as a model. EO was obtained using a hydrodistillation Clevenger-type apparatus and was qualitatively and quantitatively characterized using GC-MS and GC-FID, respectively. The alamar blue assay was used to assess the cytotoxic potential of EO in a panel of human cancer cell lines and human non-cancerous cells. In HepG2 cells treated with EO, YO-PRO-1/propidium iodide staining, cell cycle distribution, and reactive oxygen species (ROS) were examined. In C.B-17 SCID mice with HepG2 cell xenografts, the efficacy of the EO (20 and 40 mg/kg) was tested in vivo. GC-MS and GC-FID analyses showed germacrene D (17.65%), 1-epi-cubenol (13.21%), caryophyllene oxide (12.03%), spathulenol (11.26%), (E)-caryophyllene (7.26%), bicyclogermacrene (5.87%), and δ-elemene (4.95%) as the major constituents of G. olivacea leaf EO. In vitro cytotoxicity of EO was observed, including anti-liver cancer action with an IC50 value of 30.82 µg/mL for HepG2 cells. In HepG2 cells, EO treatment increased apoptotic cells and DNA fragmentation, without changes in ROS levels. Furthermore, the EO inhibited tumor mass in vivo by 32.8-57.9%. These findings suggest that G. olivacea leaf EO has anti-liver cancer potential.

Implementation of a MS/MS database for isoquinoline alkaloids and other annonaceous metabolites.

This data descriptor reports on the upload to a public repository (GNPS) of the IQAMDB, IsoQuinoline and Annonaceous Metabolites Data Base, comprising 320 tandem mass spectra. This project originated from our in-house collection of isoquinolines. The diversity of compounds included in this database was further extended through the contribution of two additional laboratories involved in isoquinoline alkaloids research: University of Angers and University of Manaus. The generated MS/MS data were processed and annotated on an individual basis to promote their straightforward reuse by natural product chemists interested in either the description of new isoquinoline alkaloids or the dereplication of isoquinoline-containing samples. The interest of the current repertoire for dereplication purposes has been validated based on the molecular networking of the well-investigated plant model Annona montana against the IQAMDB-implemented GNPS.

Plant-Derived Toxin Inhibitors as Potential Candidates to Complement Antivenom Treatment in Snakebite Envenomations.

Snakebite envenomations (SBEs) are a neglected medical condition of global importance that mainly affect the tropical and subtropical regions. Clinical manifestations include pain, edema, hemorrhage, tissue necrosis, and neurotoxic signs, and may evolve to functional loss of the affected limb, acute renal and/or respiratory failure, and even death. The standard treatment for snake envenomations is antivenom, which is produced from the hyperimmunization of animals with snake toxins. The inhibition of the effects of SBEs using natural or synthetic compounds has been suggested as a complementary treatment particularly before admission to hospital for antivenom treatment, since these alternative molecules are also able to inhibit toxins. Biodiversity-derived molecules, namely those extracted from medicinal plants, are promising sources of toxin inhibitors that can minimize the deleterious consequences of SBEs. In this review, we systematically synthesize the literature on plant metabolites that can be used as toxin-inhibiting agents, as well as present the potential mechanisms of action of molecules derived from natural sources. These findings aim to further our understanding of the potential of natural products and provide new lead compounds as auxiliary therapies for SBEs.

Essential oil of Piper purusanum C.DC (Piperaceae) and its main sesquiterpenes: biodefensives against malaria and dengue vectors, without lethal effect on non-target aquatic fauna.

The mosquito vectors of the genera Aedes and Anopheles present resistance to several commercial insecticides, which are also toxic to non-predator targets. On the other hand, essential oils are a promising source of insecticides. Thus, in this work, the essential oil from the leaves of Piper purusanum was characterized by gas chromatography-based approaches and evaluated as biodefensive against malaria and dengue vectors. The main compounds of P. purusanum essential oil were ß-caryophyllene (57.05%), α-humulene (14.50%), and germacrene D (8.20%). The essential oil inhibited egg hatching (7.6 ± 1.5 to 95.6 ± 4.5%), caused larval death (LC50 from 49.84 to 51.60 ppm), and inhibited the action of acetylcholinesterase (IC50 of 2.29 µg/mL), which can be related to the mechanisms of action. On the other hand, the biological activities of ß-caryophyllene, α-humulene, and germacrene D were higher than that of essential oil. In addition, these sesquiterpenes and essential oil did not show a lethal effect on Toxorhynchites splendens, Anisops bouvieri, Gambusia affinis, and Diplonychus indicus (LC50 from 2098.80 to 7707.13 ppm), although D. indicus is more sensitive (SI/PSF from 48.56 to 252.02 ppm) to essential oil, representing a natural alternative against these relevant vectors.

Tingenone and 22-hydroxytingenone target oxidative stress through downregulation of thioredoxin, leading to DNA double-strand break and JNK/p38-mediated apoptosis in acute myeloid leukemia HL-60 cells.

Acute myeloid leukemia (AML) is the most lethal form of leukemia. Standard anti-AML treatment remains almost unchanged for decades. Tingenone (TG) and 22-hydroxytingenone (22-HTG) are quinonemethide triterpenes found in the Amazonian plant Salacia impressifolia (Celastraceae), with cytotoxic properties in different histological types of cancer cells. In the present work, we investigated the anti-AML action mechanism of TG and 22-HTG in the AML HL-60 cell line. Both compounds exhibited potent cytotoxicity in a panel of cancer cell lines. Mechanistic studies found that TG and 22-HTG reduced cell growth and caused the externalization of phosphatidylserine, the fragmentation of internucleosomal DNA and the loss of mitochondrial transmembrane potential in HL-60 cells. In addition, pre-incubation with Z-VAD(OMe)-FMK, a pan-caspase inhibitor, prevented TG- and 22-HTG-induced apoptosis, indicating cell death by apoptosis via a caspase-dependent pathway. The analysis of the RNA transcripts of several genes indicated the interruption of the cellular antioxidant system, including the downregulation of thioredoxin, as a target for TG and 22-HTG. The application of N-acetyl-cysteine, an antioxidant, completely prevented apoptosis induced by TG and 22-HTG, indicating activation of the apoptosis pathway mediated by oxidative stress. Moreover, TG and 22-HTG induced DNA double-strand break and phosphorylation of JNK2 (T183/Y185) and p38α (T180/Y182), and co-incubation with SP 600125 (JNK/SAPK inhibitor) and PD 169316 (p38 MAPK inhibitor) partially prevented apoptosis induced by TG and 22-HTG. Together, these data indicate that TG and 22-HTG are new candidate for anti-AML therapy targeting thioredoxin.

Benzylated Dihydroflavones and Isoquinoline-Derived Alkaloids from the Bark of Diclinanona calycina (Annonaceae) and Their Cytotoxicities.

Diclinanona calycina R. E. Fries popularly known as "envira", is a species of the Annonaceae family endemic to Brazil. In our ongoing search for bioactive compounds from Annonaceae Amazon plants, the bark of D. calycina was investigated by classical chromatography techniques that yielded thirteen compounds (alkaloids and flavonoids) described for the first time in D. calycina as well as in the genus Diclinanona. The structure of these isolated compounds were established by extensive analysis using 1D/2D-NMR spectroscopy in combination with MS. The isolated alkaloids were identified as belonging to the subclasses: simple isoquinoline, thalifoline (1); aporphine, anonaine (2); oxoaporphine, liriodenine (3); benzyltetrahydroisoquinolines, (S)-(+)-reticuline (4); dehydro-oxonorreticuline (3,4-dihydro-7-hydroxy-6-methoxy-1-isoquinolinyl)(3-hydroxy-4-methoxyphenyl)-methanone) (5); (+)-1S,2R-reticuline Nß-oxide (6); and (+)-1S,2S-reticuline Nα-oxide (7); tetrahydroprotoberberine, coreximine (8); and pavine, bisnorargemonine (9). While the flavonoids belong to the benzylated dihydroflavones, isochamanetin (10), dichamanetin (11), and a mixture of uvarinol (12) and isouvarinol (13). Compound 5 is described for the first time in the literature as a natural product. The cytotoxic activity of the main isolated compounds was evaluated against cancer and non-cancerous cell lines. Among the tested compounds, the most promising results were found for the benzylated dihydroflavones dichamanetin (10), and the mixture of uvarinol (12) and isouvarinol (13), which presented moderate cytotoxic activity against the tested cancer cell lines (<20.0 µg·mL-1) and low cytotoxicity against the non-cancerous cell line MRC-5 (>25.0 µg·mL-1). Dichamanetin (11) showed cytotoxic activity against HL-60 and HCT116 with IC50 values of 15.78 µg·mL-1 (33.70 µmol·L-1) and 18.99 µg·mL-1 (40.56 µmol·L-1), respectively while the mixture of uvarinol (12) and isouvarinol (13) demonstrated cytotoxic activity against HL-60, with an IC50 value of 9.74 µg·mL-1, and HCT116, with an IC50 value of 17.31 µg·mL-1. These cytotoxic activities can be attributed to the presence of one or more hydroxybenzyl groups present in these molecules as well as the position in which these groups are linked. The cytotoxic activities of reticuline, anonaine and liriodenine have been previously established, with liriodenine being the most potent compound.

Essential Oil from Bark of Aniba parviflora (Meisn.) Mez (Lauraceae) Reduces HepG2 Cell Proliferation and Inhibits Tumor Development in a Xenograft Model.

Aniba parviflora (Meisn.) Mez (Lauraceae) is an aromatic plant of the Amazon rainforest, which has a tremendous commercial value in the perfumery industry; it is popularly used as flavoring sachets and aromatic baths. In Brazilian folk medicine, A. parviflora is used to treat victims of snakebites. Herein, we analyzed the chemical composition of A. parviflora bark essential oil (EO) and its effect on the growth of human hepatocellular carcinoma HepG2 cells in vitro and in vivo. EO was obtained by hydrodistillation and characterized by GC-MS and GC-FID. The main constituents of EO were linalool (16.3±3.15), α-humulene (14.5±2.41 %), δ-cadinene (10.2±1.09 %), α-copaene (9.51±1.12 %) and germacrene B (7.58±2.15 %). Initially, EO's cytotoxic effect was evaluated against five cancer cell lines (HepG2, MCF-7, HCT116, HL-60 and B16-F10) and one non-cancerous one (MRC-5), using the Alamar blue method after 72 h of treatment. The calculated IC50 values were 9.05, 22.04, >50, 15.36, 17.57, and 30.46 µg/mL, respectively. The best selectivity was for HepG2 cells with a selective index of 3.4. DNA Fragmentation and cell cycle distribution were quantified in HepG2 cells by flow cytometry after a treatment period of 24 and 48 h. The effect of EO on tumor development in vivo was evaluated in a xenograft model using C.B-17 SCID mice engrafted with HepG2 cells. In vivo tumor growth inhibition of HepG2 xenograft at the doses of 40 and 80 mg/kg were 12.1 and 62.4 %, respectively.

Comparative evaluation of chemical composition and biological activities of tropical fruits consumed in Manaus, central Amazonia, Brazil.

Fruits are widely recognized as sources of biologically active metabolites, such as antioxidant compounds. In this context, fruits commonly consumed in the central Amazonia, especially in its biggest metropolis (Manaus - AM/Brazil), are attractive as potential sources of antioxidant compounds related to biological activities. Most of such fruits are still poorly studied and/or remain unknown outside the Amazon region. Therefore, this study aims to investigate nine fruits (abiu, cubiu, biribá, breadfruit, genipap, peach palm, murici, soursop, and umari) regarding their chemical composition (fixed and volatile), reducing capacity, antioxidant activity, enzyme inhibition, and cytotoxicity. Determination of small organic acids, hydroxycinnamic acids, flavan-3-ols and flavonoid aglycones was done by HPLC-MS/MS, whereas determination of volatile organic compounds (VOCs) was done by HS-SPME/GC-MS. Reducing capacity was determined by the Folin-Ciocalteu method, and antioxidant activities were evaluated by DPPH, ABTS, and H-ORACFL assays. In vitro activities regarding inhibition of enzymes were tested for α-glucosidase, lipase, and α-amylase, and anti-glycation activities were evaluated for methylglyoxal and fructose. Cytotoxicity of fruit extracts was evaluated by cell viability of human fibroblast cell line (MRC-5). A total of 16 antioxidant compounds and 139 VOCs were determined, whose profiles were unique for each studied fruit. Total phenolic contents as well as antioxidant activities found herein were similar or even higher than those reported for several traditional fruits. Some of fruit extracts were able to inhibit α-glucosidase and glycation in methylglyoxal and fructose models, whereas none of them was active for lipase and α-amylase. All of the fruit extracts showed to be non-cytotoxic to MRC-5 cell line.