In Vitro Antiglycation Potential of Erva-Baleeira (Varronia curassavica Jacq.).

BACKGROUND: Varronia curassavica Jacq. (Boraginaceae) is traditionally used in the treatment of inflammatory processes. The ethanolic extract of its leaves (EEVc) showed anti-inflammatory properties and low toxicity. Medicinal plants have aroused interest for their antiglycation activities. The formation and accumulation of advanced glycation end products (AGEs) are associated with several chronic diseases. The objective of this study was to evaluate the antiglycation potential of EEVc and two isolated compounds. METHODS: The compounds brickellin and cordialin A were obtained by chromatographic methods and identified by spectrometric techniques. Analysis of fluorescent AGEs, biomarkers of amino acid residue oxidation, protein carbonyl groups and crosslink formation were performed in samples obtained from an in vitro model system of protein glycation with methylglyoxal. RESULTS: EEVc, brickellin and cordialin A significantly reduced the in vitro formation of AGEs, and reduced the damage caused by oxidative damage to the protein. CONCLUSIONS: According to the results, EEVc, brickellin and cordialin A are potential candidates against AGEs formation, which opens the way to expand the therapeutic arsenal for many pathologies resulting from glycoxidative stress.

Review: Role of the chemical compounds present in the essential oil and in the extract of Cordia verbenacea DC as an anti-inflammatory, antimicrobial and healing product.

Cordia verbenacea DC is a plant whose geographical distribution is all over Brazil, but mainly on the country's coastal areas. It is used by folk medicine to treat inflammations and chronic pain with application of the raw extract of macerated leaves in the region of pain. This paper aims to provide an integrative review of the role of chemical compounds present in the extract and the essential oil of Cordia verbenacea DC for their anti-inflammatory, antimicrobial and healing properties. The extract of Cordia verbenacea DC is widely found within popular culture as teas, decoctions, infusions, pomades/creams for anti-inflammatory, antimicrobial and healing uses. Its extract is mainly composed of α-humulene, α-pinene, trans-caryophyllene, artemetin, spathulenol and allo-aromadrendene. The described effects of the chemical compounds found in the extract are applied to muscle bruising and myofascial pain, edema reduction and chronic tendonitis, when used topically (with a comercial product being marketed); and to rheumatic, neuralgic and arthritic processes, when used orally as an anti-ulcerative agent; as well as for its healing and analgesic properties when used topically or as infusion according to test results using animals. Besides its use in chronic pain and topical application, in vitro studies have revealed the importance of antimicrobial action by inhibiting the bacterial growth of the components present in the Cordia verbenacea DC extract. In addition to expanding knowledge about Cordia verbenacea, other studies are being developed with the extract and essential oil in order to expand knowledge about healing aspects of topical use. Although other studies are being conducted, the results presented in this analysis provide a rich database of the activities found in the compounds found in the extract and the essential oil, in addition to promoting knowledge of the best extraction method cited. Therefore, such studies constitute a preliminary source of information to encourage new pieces of research, species conservation, safe management of the species, use of the extract/essential oil and development of plant-based products.

Pterodon pubescens and Cordia verbenacea association promotes a synergistic response in antinociceptive model and improves the anti-inflammatory results in animal models.

This study evaluated the pharmacological effect of the association of crude extract from the fruits of Pterodon pubescens (Pp) with the essential oil of Cordia verbenacea (Cv) in antinociception and anti-inflammatory experimental models. The effective doses of each extract and the combinations used in the associations of extracts were defined by acetic acid-induced writhing test. The separate extracts were also evaluated on formalin test. Interaction between extracts was assessed by isobologram method. The effects of different concentrations of associations (A50, A100 and A200) were evaluated on formalin test, tail flick and hot plate. The anti-inflammatory activity was evaluated in paw edema induced by carrageenan and PGE2, carrageenan-induced peritonitis and mechanical allodynia induced by Complete Freund's Adjuvant (CFA). The associations were markedly synergistic, as assessed using isobolographic analyses. On formalin and on acetic acid-induced writhing tests, associations demonstrated greater efficacy when compared to extracts separately. In paw edema models, significant reductions of edema were observed. On mechanical allodynia induced by CFA, associations were effective at acute phase with pronounced effect at chronic phase. The associations were not effective in hot plate, tail flick and carrageenan-induced peritonitis tests. Phytochemical analysis by HPLC-DAD and FID showed important concentrations of α-Humulene, trans-Caryophyllene, geranylgeraniol, isomers 6α-hydroxy-7ß-acetoxy-vouacapan-17ßoate methyl ester and 6α-acetoxy-7ß-hydroxy-vouacapan-17ß-oate methyl ester (compounds m/z 404) and 6α,7ß-dihydroxyvouacapan-17ß-oate methyl ester (m/z 362). These findings demonstrate that the associations promote synergistic antinociceptive effect and important anti-inflammatory activities, especially on chronic inflammation conditions, at lower doses than the separate crude extracts, without demonstrating side effects, probably acting in different pharmacological receptors. The inhibition of inflammation suggests a relationship with inflammatory mediators and PGE2 pathway and might be exploited to achieve greater anti-inflammatory efficacy, being considered as a potential phytotherapy.

Quali-quantitative characterization of the volatile constituents in Cordia verbenacea D.C. essential oil exploiting advanced chromatographic approaches and nuclear magnetic resonance analysis.

Cordia verbenacea D.C. (Boraginaceae, Varronia curassavica Jacq. synonym) is a medicinal plant, native from Brazil, especially the leaves are used in folk medicine. The aim of this study was to extend the characterization of the volatile fraction of the essential oil obtained from this plant, by using GC-FID, GC-MS, and chiral GC. Moreover, to further clarify the composition of the volatile fraction, preparative multidimensional-GC (prep-MDGC) was used to collect unknown compounds, followed by NMR characterization. Specifically, the chemical characterization, both qualitative and quantitative, of the volatile fraction of the essential oil obtained from Cordia verbenacea cultivated in the Minas Gerais area (central area of Brazil) was investigated for the first time. The principal components from a quantitative point of view were α-pinene (25.32%; 24.48g/100g) and α-santalene (17.90%; 17.30g/100g), belonging to the terpenes family. Chiral-GC data are reported for the enantiomeric distribution of 7 different components. Last, to obtain the complete characterization of the essential oil constituents, prep-MDGC analysis was used to attain the isolation of two compounds, not present in the principal MS databases, which were unambiguously identified by NMR investigation as (E)-α-santalal and (E)-α-bergamotenal, reported for the first time in Cordia verbenacea essential oil.

Potentiation of antibiotic activity of aminoglycosides by natural products from Cordia verbenacea DC.

Medicinal plants are often the only therapeutic resource for many communities and ethnic groups. Cordia verbenacea DC., "Erva-baleeira," is one of the species of plants currently used to produce a phytotherapeutic product extracted from its leaves. The present study aimed to establish its chemical profile, antibacterial activity and resistance-modulating potential. The C. verbenacea extracts were prepared from fresh leaves using solvents as methanol and hexane. Ethyl Acetate was used for the preparation of the fraction. Phytochemical screening was carried out using HPLC-DAD for determination and quantification of the secondary metabolites present in the fractions. Antibacterial and resistance-modulation assays were performed to determine minimum inhibitory concentration (MIC) using a microdilution assay. The data were subjected to statistical analysis with two-way ANOVA and Bonferroni posttests. Results of phytochemical prospecting and HPLC analysis of the fractions were in agreement with the literature. The natural products presented moderate antibacterial activity when considering the clinical relevance of a MIC of 256 µg/mL against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa, and 512 µg/mL against P. aeruginosa. However, when the fractions were combined with antibiotics we observed a synergic effect, as natural products enhanced the antibacterial effect of aminoglycosides, significantly decreasing the MIC of antibiotics at 12.5%-98.4%. We believe that the data obtained from phytochemical analysis and from antibacterial and resistance modulation assays of C. verbenacea extracts new can open perspectives in the search for new alternatives for the treatment of bacterial infections and stimulate the renewed use of antibiotics with reduced effectiveness due to resistance.

Medicinal plants in Brazil: Pharmacological studies, drug discovery, challenges and perspectives.

This review article focuses on pre-clinical and clinical studies with some selected Brazilian medicinal plants in different areas of interest, conducted by research groups in Brazil and abroad. It also highlights the Brazilian market of herbal products and the efforts of Brazilian scientists to develop new phytomedicines. This review is divided into three sections. The section I describes the Brazilian large biodiversity and some attempts of Brazilian scientists to assess the pharmacological profile of most plant extracts or isolated active principles. Of note, Brazilian scientists have made a great effort to study the Brazilian biodiversity, especially among the higher plants. In fact, more than 10,000 papers were published on plants in international scientific journals between 2011 and 2013. This first part also discussed the main efforts to develop new medicines from plants, highlighting the Brazilian phytomedicines market. Despite the large Brazilian biodiversity, notably with the higher plants, which comprise over 45,000 species (20-22% of the total worldwide), and the substantial number of scientific publications on medicinal plants, only one phytomedicine is found in the top 20 market products. Indeed, this market is still only worth about 261 million American dollars. This represents less than 5% of the global Brazilian medicine market. The section II of this review focus on the use of Brazilian plant extract and/or active principles for some selected diseases, namely: central nervous systems disorders, pain, immune response and inflammation, respiratory diseases, gastrointestinal tract and metabolic diseases. Finally, section III discusses in more details some selected Brazilian medicinal plants including: Cordia verbenacea, Euphorbia tirucalli, Mandevilla velutina, Phyllanthus spp., Euterpe oleracea, Vitis labrusca, Hypericum caprifoliatum and Hypericum polyanthemum, Maytenus ilicifolia, Protium kleinii and Protium heptaphylium and Trichilia catigua. Most of these publications are preliminary and only report the effects of crude extracts, both in vitro and in vivo studies. Only very few studies have been dedicated to investigate the mechanisms of action of isolated compounds. Likewise, studies on safety (toxicology), pharmacokinetic, and especially on well-conducted clinical trials are rare. In conclusion, in spite of the abundant Brazilian biodiversity and the thousands of academic publications on plants in international peer-reviewed scientific journals, few patents and medicines have been derived from such studies. Undoubtedly, great efforts must be made to improve the development of plant-derived medicine market in Brazil, especially by involving the partnership between academia and pharmaceutical companies.

Determinação de ácido rosmarínico em Cordia verbenacea por cromatografia líquida: aplicabilidade em estudo sazonal / Determination of rosmarinic acid in Cordia verbenacea by liquid chromatography: applicability in seasonal study

RESUMO Neste estudo, uma técnica de cromatografia líquida de alta resolução em fase reversa (CLAE-FR) para a determinação de ácido rosmarínico em Cordia verbenacea foi desenvolvida e validada. A análise de regressão foi avaliada, com observação de uma boa linearidade (r = 0,9997). Os valores obtidos para a precisão e exatidão estão de acordo com as diretrizes do ICH e com a legislação brasileira. Os valores de repetibilidade e precisão intermediária foram 2,79% e 4,76%, respectivamente. Os limites de detecção e de quantificação de ácido rosmarínico foram de 1,92 µg/mL e 5,81 µg/mL, respectivamente. Os resultados mostraram que o método desenvolvido é uma técnica por CLAE-FR de confiança para a determinação de ácido rosmarínico em tintura de C. verbenacea. Além disso, essa metodologia foi aplicada em estudo sazonal, que revela uma correlação positiva relativamente forte entre o período de chuvas e o teor de ácido rosmarínico.

ABSTRACT In this study, a reverse phase-high performance liquid chromatography (RP-HPLC) technique for determination of rosmarinic acid in the Cordia verbenacea was developed and validated. A regression analysis was performed, with the observation of good linearity (r =0.999949). The values obtained for precision and accuracy determination are in agreement with ICH guidelines and the Brazilian legislation. The values of repeatability and intermediate precision were 2.79% and 4.76%, respectively. The detection and the quantitation limits of the rosmarinic acid were 1.92 µg/mL and 5.81 µg/mL, respectively. The results demonstrated that the developed method is a reliable RP-HPLC technique for the determination of rosmarinic acid in C. verbenacea tincture. In addition, this methodology was applied at a seasonal study indicating relatively strong positive correlation between the rain period and the rosmarinic acid content.

Light affects Varronia curassavica essential oil yield by increasing trichomes frequency

Light can act on essential oil yield directly on synthesis of secondary metabolites, or indirectly on plant growth. Varronia curassavica Jacq., Boraginaceae, is a native medicinal species from Brazil known as “erva-baleeira”, with anti-inflammatory activity related to its essential oil. Despite pharmacological evidences of this species and its economic importance for herbal medicine production, little is known about the effect of light on growth and essential oil production. This study aimed to analyze the influence of different irradiances on growth, frequency of trichomes, essential oil yield and composition of V. curassavica. The irradiance affected plant growth, but no significant alteration on leaf biomass was detected. The increase in essential oil content under higher irradiance reflected on essential oil yield, and is associated with higher frequency of glandular, globular trichomes. The essential oil composition, rich in caryophyllene derivatives was affected by irradiance, but α-humulene, the constituent of pharmaceutical interest, remained unchanged.

Determinação do perfil fitoquímico de extrato com atividade antioxidante da espécie medicinal Cordia verbenacea DC. por HPLC-DAD / Phytochemical profile determination from extracts with antioxidant activity of the medicinal species of Cordia verbenacea DC. by HPLC-DAD

O presente trabalho teve por objetivo investigar a atividade antioxidante dos extratos das folhas de Cordia verbenacea obtido por maceração em etanol e partição em solventes orgânicos. O infuso das folhas também foi investigado. O teor de fenóis totais foi avaliado pelo método de Folin-Ciocalteau e o de flavonoides totais pela formação de complexo com cloreto de alumínio. O extrato etanólico, as subfrações e o infuso foram testados em diversas concentrações para determinar a atividade sequestradora de DPPH expressa em termos de sua CE50. A melhor atividade antioxidante encontrada foi para o extrato em acetato de etila, EA, CE50 15,0 ± 0,5 µg.mL-1. Os ensaios espectrofotométricos revelaram altas concentrações de fenóis e de flavonoides no extrato EA. A análise por HPLC-DAD foi realizada para se obter o perfil de UV-Vis dos picos cromatográficos do extrato EA. As características espectrais foram relacionadas a compostos fenólicos e flavonoídicos.

The objective of the present study was to investigate the antioxidant activity of extracts from Cordia verbenacea leaves obtained by maceration in ethanol and partitioned with organic solvents. The infusion of leaves was also investigated. The total phenolic and total flavonoid content was evaluated by the Folin-Ciocalteau method and by the aluminum chloride complex method, respectively. The ethanol extract, the subfractions and the infusion were assayed at various concentrations to determine the DPPH scavenging activity expressed in terms of CE50. The best antioxidant activity was found on the ethyl acetate extract, EA, CE50 15.0 ± 0.5 µg.mL-1. The spectrophotometric assays revealed high phenol and flavonoid concentrations on the EA extract. The HPLC-DAD analysis was performed to reveal the UV-Vis profile of the chromatographic peaks of the EA extract. The UV spectral characteristics were related to phenol and polyphenol compounds.

Chemical composition, antibacterial and antifungal activities of essential oil from Cordia verbenacea DC leaves.

BACKGROUND: Cordia verbenacea is a Brazilian coastal shrub popularly known as "erva baleeira". The essential oil from fresh leaves was obtained by hydrodistillation and analyzed by CG/MS. The main components were identified as ß-caryophyllene (25.4%), bicyclogermacrene (11.3%), δ-cadinene (9.%) and α-pinene (9.5%). In this study, the antimicrobial activity of Cordia verbenacea was evaluated. MATERIALS AND METHODS: The minimal inhibitory concentration (MIC) of the essential oil was obtained using the broth microdilution assay (from 512 to 8 µg/ml). RESULTS: The results showed that the essential oil presented fungistatic activity against Candida albicans and Candida krusei and antibacterial activity against Gram-positive strains (Staphylococcus aureus and Bacillus cereus) and against multiresistant Gram-negative (Escherichia coli 27), in all tests the MIC was 64 µg/ml. When the essential oil was associated to aminoglycosides (subinhibitory concentrations, MIC/8), a synergic and antagonic activity was verified. The synergic effect was observed to the amikacin association (MIC reduction from 256 mlto 64 µg/ml) in all strains tested. CONCLUSION: The essential oil of Cordia verbenacea influences the activity of antibiotics and may be used as an adjuvant in antibiotic therapy against respiratory tract bacterial pathogens.