A neolignan from Connarus tuberosus as an allosteric GABAA receptor modulator at the neurosteroid binding site.

In a screening of a small library of extracts from plants of the Amazonian and Cerrado biomes, a hexane extract of Connarus tuberosus roots was found to significantly potentiate the GABA induced fluorescence in a fluorescence (FLIPR) assay in CHO cells stably expressing the α1ß2γ2 subtype of human GABAA receptors. With the aid of HPLC-based activity profiling the activity was linked to the neolignan connarin. In CHO cells the activity of connarin was not abolished by increasing concentrations of flumazenil, while the effect of diazepam was increased by increasing concentrations of connarin. The effect of connarin was abolished by pregnenolone sulfate (PREGS) in a concentration-dependent manner, and the effect of allopregnanolone was further increased by increasing concentrations of connarin. In a two-microelectrode voltage clamp assay with Xenopus laevis oocytes transiently expressing GABAA receptors composed of human α1ß2γ2S and α1ß2 subunits connarin potentiated the GABA-induced currents, with EC50 values of 1.2 ± 0.3 µM (α1ß2γ2S) and 1.3 ± 0.4 µM (α1ß2), and with a maximum enhancement of currents Emax of 1959 ± 70% (α1ß2γ2S) and 185 ± 48% (α1ß2). The activation induced by connarin was abolished by increasing concentrations of PREGS.

Chemical constituents from fruits of Cnestis ferruginea Vahl ex. DC (Connaraceae) and evaluation of their anticholinesterase and antiradical activities.

The phytochemical investigation of the DCM/MeOH (1:1) extract of the fruits of Cnestis ferruginea led to the isolation and characterization of one new quinic acid derivative, ferruginoic acid (1), together with six known compounds 2-7. Compounds 3-7 were reported for the first time from this species. The structures of compounds 1-7 were elucidated on the basis of 1 D and 2 D NMR spectroscopic data, mass spectrometry and by comparison of spectroscopic data with those from the literature. The anticholinesterase (AChE and BChE) activity and DPPH free radical scavenging assay of compounds 1, 3, 4 and 7 were evaluated. Ferruginoic acid (1) exhibited moderate anticholinesterase activity with IC50 value of 36.18 ± 1.78 µg/mL against AChE. Compounds 3, 4 and 7 showed high activity against free radical (DPPH•) scavenging assay (DPPH) with IC50 values 40.09 ± 0.96 µg/mL, 61.70 ± 0.78 µg/mL and 41.87 ± 0.62 µg/mL respectively. These results indicate that C. ferruginea and its constituents could be employed in the management of Alzheimer's disease.

Four almost unexplored species of Brazilian Connarus (Connaraceae): Chemical composition by ESI-QTof-MS/MS-GNPS and a pharmacologic potential.

INTRODUCTION: Species of Connaraceae are globally used in traditional medicines. However, several of these have not been studied regarding their chemical composition, and some are even at risk of extinction without proper studies. Therefore, the chemical composition and pharmacological potential of Connarus blanchetii Planch., Connarus nodosus Baker, Connarus regnellii G. Schellenb., and Connarus suberosus Planch., which were previously unknown, were analyzed. OBJECTIVE: This work aims to investigate the pharmacological potential of these four Connarus species. The chemical composition of different extracts was determined by high-resolution mass spectrometry (HRMS), with subsequent analysis by the GNPS platform and competitive fragmentation modeling (CFM). MATERIALS AND METHODS: Leaf extracts (C. blanchetii, C. nodosus, C. regnellii, and C. suberosus) and bark extracts (C. regnellii and C. suberosus) were obtained by decoction, infusion, and maceration. LC/HRMS data were submitted to the GNPS platform and evaluated using CFM in order to confirm the structures. RESULTS: The HRMS-GNPS/CFM analysis indicated the presence of 23 compounds that were mainly identified as phenolic derivatives from quercetin and myricetin, of which 21 are unedited in the Connarus genus. Thus, from the analyses performed, we can identify different compounds with pharmacological potential, as well as the most suitable forms of extraction. CONCLUSION: Using HRMS-GNPS/CFM, 21 unpublished compounds were identified in the studied species. Therefore, our combination of data analysis techniques can be used to determine their chemical composition.

Connaraceae: An updated overview of research and the pharmacological potential of 39 species.

ETHNOPHARMACOLOGICAL RELEVANCE: An interdisciplinary scientific investigation of biologically active agents is fundamental to search for natural substances with therapeutic action. This review collected the most relevant information on traditional knowledge related to the use of plants of the Connaraceae family. This work is the first to compile all the published ethnobotanical, chemical, pharmacological, and toxicological information about this important plant family. AIM OF THE STUDY: Our objective was to provide the scientific community with an up-to-date overview of the pharmacological potential of Connaraceae species. MATERIAL AND METHODS: We searched NCBI Pubmed Central, Google Scholar, Scientific Electronic Library Online (SciELO), ScienceDirect, SciFinder, and Scopus databases to review the research on ethnobotanical, chemical, pharmacognostical, pharmacological, and toxicological studies with Connaraceaes. Books that address the theme were also included. DISCUSSION AND CONCLUSION: The literature review indicated that 39 species of Connaraceaes have pharmacological potentiality. Ethnobotany reports listed 36 of the 39 species discussed. Pharmacognostical studies have been conducted with 23 species and isolates, and chemical compounds have been identified for only 15 species. At least one study has been published concerning the pharmacological activities for 20 of the 39 species analyzed. For Agelaea pentagyna, Cnestis ferruginea, Connars suberosus, and Rourea minor, pharmacological activity experiments were performed using isolated compounds, which have the highest current pharmacological potential. Studies employing a toxicological approach cover only 10 of the 39 Connaraceaes species. Thus, scientific community should conduct much more research for a broader understanding of this plant family.

Brysocarpus coccineus (Schum & Thonn) root reinstates sexual competence and testicular function in paroxetine-induced sexual dysfunction in male Wistar rats.

The effects of aqueous extract of Brysocarpus coccineus roots (AEBCR) were studied on sexual behaviour and testicular function of paroxetine-induced sexual dysfunction (SD) in male rats. Ninety, sexually matured male rats (150.88 ± 5.53 g) were assigned into two groups: A and B. Fifteen SD animals from group B were each allotted to B1, B2, B3, B4 and B5 and received distilled water (DW), Powmax M (7.14 mg/kg body weight, b.w.) 50, 100 and 150 mg/kg b.w of AEBCR, respectively, for 7 days while the non-SD animals (group A) received DW. Eleven secondary metabolites were present in AEBCR. The lowered (p < .05) ejaculation frequency, penile erection index and penile grooming, higher mount and intromission frequencies, prolonged (p < .05) latencies of mount, intromission, ejaculation, and post-ejaculatory interval, reduced (p < .05) serum luteinising hormone, follicle stimulating hormone, testosterone, nitric oxide and testicular function indices, degenerated seminiferous tubules and low luminal spermatozoa contents by paroxetine were significantly (p < .05) attenuated and/or reinstated by AEBCR and Powmax M. The restoration of androgen-dependent sexual and testicular functions in SD male rats by AEBCR validates its folkloric use as aphrodisiac. Clinical studies are desirable to ascertain the efficacy of AEBCR in SD.

Ameliorative Effect of Hydroethanolic Leaf Extract of Byrsocarpus coccineus in Alcohol- and Sucrose-Induced Hypertension in Rats.

Hypertension remains a major health problem worldwide considering the prevalence of morbidity and mortality. Plants remain a reliable source of efficacious and better tolerated drugs and botanicals. This study was designed to investigate the effect of the chemo-profiled hydroethanolic leaf extract of Byrsocarpus coccineus in ethanol- and sucrose-induced hypertension. Groups of rats were treated orally (p.o.) with distilled water (10 ml/kg), ethanol (35%; 3 g/kg), sucrose (5-7%), and B. coccineus (100, 200, and 400 mg/kg), and nifedipine together with ethanol and sucrose separately for 8 weeks. At the end of the treatment period, blood pressure and heart rate of rats were determined. Blood was collected for serum biochemical parameters and lipid profile assessment, and the liver, aorta, kidney, and heart were harvested for estimation of in vivo antioxidants and malondialdehyde (MDA). Results obtained in this study showed that B. coccineus at the various doses administered reduced the systolic, diastolic, and arterial blood pressure elevated by ethanol and sucrose. Also, the extract reversed the reduction in catalase (CAT), reduced glutathione (GSH), glutathione peroxidase (GPx), and superoxide dismutase (SOD) induced by ethanol and sucrose. The level of MDA was reduced compared to the ethanol- and sucrose-induced hypertensive group. With respect to lipid profile, administration of B. coccineus at the various doses reduced the levels of triglycerides, low-density lipoprotein (LDL), cholesterol, and atherogenic indices, compared to the ethanol and sucrose groups. In conclusion the hydroethanolic leaf extract of B. coccineus exerted significant antihypertensive effect and this is probably related to the antioxidant property and improvement of lipid profile observed in this study.

Chemical composition and antinociceptive effect of aqueous extract from Rourea induta Planch. leaves in acute and chronic pain models.

ETHNOPHARMACOLOGICAL RELEVANCE: Rourea induta Planch. is a small tree or shrub growing wild in Brazil and belonging to the Connaraceae family. It is used for the treatment of Chagas disease and as antirheumatic in folk medicine. This study was designed to investigate the antinociceptive activity of the aqueous extract (AERi) of Rourea induta leaves in rodents, as well as isolate and identify components that can be responsible for its effect. MATERIAL AND METHODS: The antinociceptive effect of orally administered AERi was evaluated in behavioral models of acute (formalin) and chronic [complete Freund׳s adjuvant (CFA)] pain in mice. We also investigated the possible involvement of opioid receptors and proinflammatory cytokines (interleukin-1ß and tumor necrosis factor-α) in the antinociceptive effect of the AERi. In addition, a nonspecific effect of AERi was evaluated by measuring locomotor activity and corporal temperature. Finally, we performed a phytochemical analysis of AERi. RESULTS: HPLC titration revealed the presence of hyperin (21.6 mg/g), quercetin-3-O-ß-xyloside (4.4 mg/g), quercetin-3-O-α-arabinofuranoside (12.0 mg/g), and quercetin (2.1 mg/g). It was also possible to isolate minor constituents׳ chlorogenic acid, neochlorogenic acid and procyanidin C1. The oral administration of AERi (100 mg/kg) significantly inhibited the neurogenic (37 ± 5%) and inflammatory (34 ± 7%) phases of formalin-induced pain. Acute and repeated treatment of animals with AERi (100 mg/kg, p.o.) once a day markedly reduced the mechanical hypersensitivity response induced by CFA, and this effect was evident until the day 10. Moreover, repeated treatment with AERi (100 mg/kg, p.o.) significantly reduced the levels of IL-1ß and TNF-α in the paw (22 ± 7% and 50 ± 19%) and in the spinal cord (100% and 100%) when compared to the CFA group. The AERi (100 mg/kg, p.o.) markedly reduced biting behavior induced by intrathecal injection of TNF-α (81 ± 11%). Finally, the effect of AERi was not associated with changes in locomotor activity or the corporal temperature of animals. CONCLUSION: These data show that aqueous extract of Rourea induta has significant antinociceptive action, which seems to be associated with an inhibition of pro-inflammatory cytokines activated pathways. These findings support the ethnomedical uses of this plant.

Hypoglycemic and antioxidant activities of the hydroethanolic leaf extract of Byrsocarpus coccineus Schumach. & Thonn. (Connaraceae).

AIM: Diabetes Mellitus is associated with significant morbidity and mortality worldwide. The need for enhanced efficacy and safety, and cheaper and more readily available new drugs has increased the search for new antidiabetic drugs from plants. This study was conducted to investigate the antidiabetic activity of the hydroethanolic leaf extract of B. coccineus in rats. METHODS: The effect of B. coccineus extract (100-800 mg · kg(-1), p.o.) on blood glucose levels in normal and glucose loaded rats, and alloxan-induced diabetic rats was determined. After 10 days of treatment, blood samples were collected from rats for lipid and insulin profiling. Animals were thereafter sacrificed and the kidneys, heart, and liver were harvested for antioxidant indices assay. RESULTS: In normal rats, B. coccineus did not cause significant reduction in blood glucose. At the dose of 800 mg · kg(-1), significant increase in blood glucose level was not observed 30 min. after glucose load. B. coccineus administered acutely did not generally produce significant reduction in blood glucose level in diabetic rats. Administered subacutely, the extract significantly reduced blood glucose level in diabetic rats from the 3(rd) day with peak effect observed at the dose of 800 mg · kg(-1) on the 10(th) day. The extract generally preserved in vivo antioxidant levels in the kidneys, heart, and liver, increased the level of high density lipoprotein and insulin, and reduced the level of triglycerides and low density lipoprotein compared to diabetic control. CONCLUSION: The findings in this study suggest that the hydroethanolic leaf extract Byrsocarpus coccineus possesses antidiabetic activity possibly mediated through inhibition of intestinal glucose absorption, in vivo antioxidant activity, and enhancement of regeneration of beta cells of the pancreas and insulin secretion.

Chemical constituents of Rourea doniana

The chromatography fractionation of the hexane, chloroform and ethyl acetate extracts from the leaves and stems of Rourea doniana Baker, Connaraceae, resulted in the isolation of five triterpenes (lupeol, lupenone, α-amyrenone, β-amyrenone, and taraxerol), a flavonol (7,4'-dimethylkaempferol), a coumarin (scopoletin) and four phytosteroids (β-sitosterol, stigmasterol, β-sitosteryl-3-O-β- D-glucopyranoside and stigmasteryl-3-O-β-D-glucopyranoside). All compounds are being for the first time in this species and all triterpenes and the flavonol are being described for the first time in the family Connaraceae. These compounds were identified on basis of their IR and NMR (¹H, 13C, DEPT, HSQC, HMBC, and NOESY) spectral data and by comparison with literature data.

Phytochemical constituents and preliminary toxicity evaluation of leaves from Rourea induta Planch. (Connaraceae) / Constituintes fitoquímicos e avaliação preliminar de toxicidade folhas de Rourea Induta Planch. (Connaraceae)

Most active plants are toxic at high doses and it is therefore important to investigate the preliminary toxicity of plant extracts. The Rourea induta species is a potential drug with no phytochemical or biological studies registered in the literature. Thus, a phytochemical study and a toxicity analysis of the ethanolic extract obtained from the leaves of Rourea induta Planch., Connaraceae, was run. A long chain hydrocarbon, n-tetracosane, and four flavonoids were identified: quercetin, and three glycosylated derivates, quercetin-3-O-α-arabinofuranoside, quercetin-3-O-β-xyloside and quercetin-3-O-β-galactoside. This is the first time these have been isolated in this species. The structures were elucidated by 13C NMR, ¹H NMR, UV and IR spectroscopy. The toxicity evaluation of extracts was performed by the brine shrimp method and determination of hemolytic activity. The samples demonstrated no toxic potential by the analyzed methods.

A maioria das plantas ativas é tóxica em doses elevadas, portanto, é importante a investigação da toxicidade preliminar dos extratos das plantas. A espécie Rourea induta é uma droga potencial que não apresenta estudo fitoquímico ou biológico descrito na literatura. Assim, um estudo fitoquímico e análises toxicológicas foram realizados com o extrato etanólico obtido das folhas de Rourea induta Planch., Connaraceae. Foram obtidos um hidrocarboneto de cadeia longa, n-tetracosano, e quatro flavonóides, quercetina e três derivados glicosilados, quercetina-3-O-α-arabinofuranosideo, quercetina-3-O-β-xilosideo e quercetina-3-O-β-galactosideo. Esta é a primeira vez que estes compostos são isolados nesta espécie. As estruturas foram elucidadas por espectroscopia de RMN-13C, RMN-¹H, ultravioleta e infravermelho. A avaliação da toxicidade dos extratos foi analisada pelo método da Artemia salina e atividade hemolítica. Nenhuma das amostras testadas apresentou um potencial tóxico pelos métodos analisados.

Connaraceae endémicas del Perú

La familia Connaraceae es reconocida en el Perú por presentar tres géneros y 20 especies (Brako & Zarucchi, 1993; Ulloa Ulloa et al., 2004), principalmente lianas y arbustos. En este trabajo reconocemos tres especies endémicas en tres géneros. Las especies endémicas ocupan la región Bosques Húmedos Amazónicos, entre los 100 y 400 m de altitud. Una de estas especies está representada dentro del Sistema Nacional de Áreas Naturales Protegidas por el Estado.

The Connaraceae are represented in Peru by 20 species in three genera (Brako & Zarucchi, 1993; Ulloa Ulloa et al., 2004), mainly lianas and trees. Here we recognize three endemic species in three genera. Endemic species are found in Humid Lowland Amazonian Forests region, between 100 and 400 m elevation. One of these species has been recorded within Peru´s protected areas system.