Malaria is nowadays one of the most serious health concerns in a global scale and, although there is an evident increase in research studies in this area, pointed by the vast number of hits and leads, it still appears as a recurrent topic every year due to the drug resistance shown by the parasite exposing the urgent need to develop new antimalarial medications. In this work, 38 molecules were synthesized via copper(I)-catalyzed alkyne-azide cycloaddition (CuAAC) or “click” chemistry, following different routes to produce 2 different organic azides, obtained from a 4,7 dicholoquinoline, reacted with 19 different commercially available terminal alkynes. All those new compounds were evaluated for their in vitro activity against the chloroquine resistant malaria parasite Plasmodium falciparum (W2). The cytotoxicity evaluation was accomplished using Hep G2 cells and SI index was calculated for every molecule. Some of the quinoline derivatives have shown high antimalarial activity, with IC50 values in the range of 1.72-8.66 µM, low cytotoxicity, with CC50>1000 µM and selectivity index (SI) in the range of 20-100, with some compounds showing SI>800. Therefore, the quinolinotriazole hybrids could be considered a very important step on the development of new antimalarial drugs.

Resumo A espécie Cissus erosa (Vitaceae) conhecida popularmente, no Brasil, como Cipó fogo é uma planta medicinal empregada no tratamento de verrugas e úlceras externas. O presente estudo objetivou a avaliação da atividade antiviral frente a Dengue e Zika virus dos extratos etanólicos de caules e de folhas desta espécie pelo método colorimétrico do MTT e determinar a caracterização fitoquímica por cromatografia líquida de ultra eficiência acoplada a espectrometria de massa destes extratos (CLUE-EM). Os extratos apresentaram atividade antiviral e baixa citotoxicidade. O extrato de folhas apresentou CC50 > 500 µg/ml para células LLCMK2 e CE50 = 18,2 µg/mL, IS > 27,5 contra o Dengue virus 2 (DENV 2). Ambos os extratos (caules e folhas) apresentaram atividade anti-Zika virus com CE50 de 45,8 µg/ml e 82,8 µg/ml, respectivamente. Os extratos apresentaram CC50 de 309,2 µg/ml (folhas) e 387,6 µg/ml (caules) para células Vero. Nas análises por CCD e CLAE-DAD detectou-se a presença de triterpenos, esteroides, flavonoides e taninos. Análises por CLUE-EM dos extratos permitiram a identificação parcial dos flavonoides majoritárias destes extratos como sendo, vitexina, isovitexina, orientina, isoorientina, luteolina-7,4’-di-O-glicosilflavona, metoxiluteolia-6(8)-C-hexosil. Os resultados dos estudos fitoquímicos até o momento sugerem que flavonoides e terpenoides são os constituintes majoritários dos extratos etanólicos destas espécies e devem contribuírem para a atividade antiviral observada in vitro.

Abstract Cissus erosa (Vitaceae), popularly known in Brazil as Cipó-fogo, is a medicinal plant used in the treatment of warts and external ulcers. The present study aimed to evaluate the activity of stems and leaves ethanol extracts of C. erosa against the Dengue and Zika virus by the MTT colorimetric method and to carry on the phytochemical characterization of active extracts by high performance liquid chromatography coupled to mass spectrometry (UPLC-MS). Only the leaves ethanol extract showed anti-Dengue virus activity EC50 18.2 µg/ml (SI > 27.5) and low cytotoxicity for LLCMK2 cells (CC50 > 500 mg/ml). Both extracts (stems and leaves) showed anti-Zika virus activity with EC50 of 45.8 mg/ml and 82.8 mg/ml, respectively. These extracts presented CC50 of 309.2 µg/ml (leaves) and 387.6 µg/ml (stems) to Vero cells. Analysis by CCD and HPLC-DAD detected the presence of triterpenes, steroids, flavonoids and tannins. UPLC-MS analyses of these extracts, allowed the identification of the majority of flavonoids present known as vitexin, isovitexin, orientin, isoorientin and two flavones derivatives, methoxyluteolin-6(8)-C-hexosyl and luteolin-7,4’-di-O-glycosylflavone. The results of the phytochemical studies here described suggest that flavonoids and terpenoids are the substances that contribute to the antiviral activity of the ethanol extracts within this species.

Malaria is responsible for more deaths around the world than any other parasitic disease. Due to the emergence of strains that are resistant to the current chemotherapeutic antimalarial arsenal, the search for new antimalarial drugs remains urgent though hampered by a lack of knowledge regarding the molecular mechanisms of artemisinin resistance. Semisynthetic compounds derived from diterpenes from the medicinal plant Wedelia paludosawere tested in silico against the Plasmodium falciparumCa2+-ATPase, PfATP6. This protein was constructed by comparative modelling using the three-dimensional structure of a homologous protein, 1IWO, as a scaffold. Compound 21 showed the best docking scores, indicating a better interaction with PfATP6 than that of thapsigargin, the natural inhibitor. Inhibition of PfATP6 by diterpene compounds could promote a change in calcium homeostasis, leading to parasite death. These data suggest PfATP6 as a potential target for the antimalarial ent-kaurane diterpenes.

Extratos etanólicos de dezoito espécies vegetais pertencentes à família Bignoniaceae, das quais sete são descritas como de uso medicinal, foram avaliados, pelo ensaio colorimétrico do MTT, para atividades citotóxica, em células Vero, e antiviral, frente aos vírus herpes simplex-tipo 1, vaccinia e encefalomiocardite murina. A maior parte dos extratos não apresentou citotoxicidade até a concentração de 500 μg/mL. Dos 28 extratos testados quatorze (50%) apresentaram atividade antiviral com valores de CE50 na faixa de 4,6+03 a 377,2+17,7 μg/mL. Somente duas espécies, Arrabidaea samydoides e Callichlamys latifolia, foram ativas frente aos três vírus. Os extratos foram caracterizados pelos seus perfís cromatográficos em CCD e CLAE-FR. Análises por CLAE-FR mostraram que a mangiferina é o constituinte majoritário em A. samydoides mas a substância isolada foi menos ativa do que o extrato bruto. Esta é a primeira vez que se relata a atividade antiviral de extratos das dezoito espécies avaliadas.

Ethanol extracts of eighteen Bignoniaceae species have been evaluated by the MTT assay for cytotoxicity in Vero cells and for antiviral activity against Human herpes virus type 1, Vaccinia virus and murine Encephalomyocarditis virus. Among such species, seven are reported to be of traditional medicinal use No cytotoxicity was observed for most of the extracts up to the concentration of 500 μg/mL. Fourteen (50%) of the 28 extracts assayed have disclosed antiviral activity with EC50 values in the range of 4.6+0.3 to 377.2+17.7 μg/mL. Only two species, Arrabidaea samydoides and Callichlamys latifolia, have shown activity against all the three viruses. The extracts were chemically characterized by their TLC and HPLC-DAD profiles. Mangiferin is the major constituent of A. samydoides but the isolated compound has been less active than the crude extract. This is the first report on the antiviral evaluation of the eighteen Bignoniaceae species assayed.

Polygonum hydropiperoides Michaux é uma espécie conhecida popularmente como "erva-de-bicho", amplamente utilizada na medicina tradicional como anti-hemorroidal, antiinflamatória e antidiarréica. O presente trabalho tem como objetivo a caracterização anatômica e histoquímica da folha, caule e raiz, que constituem a droga vegetal, visando estabelecer parâmetros para o controle de qualidade. O material vegetal foi fixado e submetido às técnicas usuais de microscopia de luz e aos testes histoquímicos. A folha é anfiestomática, dorsiventral e com estômatos paracíticos e anisocíticos. É comum a presença de estruturas secretoras como: tricomas glandulares capitados e glândulas epidérmicas e subepidérmicas em ambas as faces da lâmina foliar e também no caule. O material secretado pelas glândulas apresenta composição heterogênea de lipídios e flavonóides, segundo análises histoquímicas. Cristais de oxalato de cálcio e grãos de amido são freqüentes em células parenquimáticas da folha e do caule. Compostos fenólicos estão presentes na folha (parênquima paliçádico), no caule (parênquima cortical e floema) e na raiz (parênquima cortical).

P. hydropiperoides is popularly known as "erva-de-bicho", and it is used in traditional medicine as anti-hemorroidal, anti-inflammatory and antidiarrhoeic. The present work has as objective the anatomical and histochemical characterization of leaf, stem and root that are used as drug in order to offer elements for quality control. The botanical material was prepared for the usual optical and histochemical microtechniques. The leaf is amphistomatic, dorsiventral and paracytic and anisocytic stomata. It is common to find the presence of secretory structures as: capitate glandular trichomes and secretory glands on both epidermis, that are also present in stem. The secretion material is heterogeneous composition of lipids and flavonoids. Druses of calcium oxalate and starch are common in parenchyma cells of leaf and stem. Phenolic compounds are present in leaf (palisade parenchyma), in stem (cortical parenchyma and phloem) and root (cortical parenchyma).

As espécies Trembleya laniflora (Melastomataceae), Xyris platystachia (Xyridaceae) e Xyris pterygoblephara foram coletadas na Serra do Cipó, região considerada hotspot para conservação de biodiversidade. A atividade antimicrobiana dessas espécies foi avaliada em ensaios in vitro de difusão em ágar frente a linhagens padronizadas de Staphylococcus aureus e Micrococcus luteus. Todos os extratos, avaliados na concentração de 2000 µg/disco, foram ativos contra M. luteus, enquanto a inibição de crescimento de S. aureus somente foi observada para os extratos de T. laniflora (folhas) e X. platystachia (partes aéreas). A partição dos extratos brutos entre solventes imiscíveis resultou na obtenção de frações ativas, oriundas de extratos originalmente inativos frente a S. aureus, observando-se atividade principalmente para as frações de baixa e média polaridade. O extrato de folhas de T. laniflora foi adicionalmente fracionado por cromatografia em coluna de sílica gel e as frações resultantes apresentaram atividade antimicrobiana e perfis por CLAE distintos daquelas obtidas pela partição entre solventes imiscíveis.

Trembleya laniflora (D. Don) Cogn. (Melastomataceae), Xyris platystachia Alb. Nilss. (Xyridaceae) and Xyris pterygoblephara Kunth., Brazilian species collected from a biodiversity hotspot for conservation priority, had their antimicrobial activity evaluated against standardized strains of Staphylococcus aureus and Micrococcus luteus, by the agar diffusion assay. All extracts, assayed in the concentration of 2000 µg/disc, were active against M. luteus, whereas S. aureus growth was inhibited only by T. laniflora leaves and X. platystachia aerial parts. Fractionation of the extracts by partition between immiscible solvents resulted in active fractions from extracts originally inactive against S. aureus. Activity was mainly found in low and medium polar fractions. The extract of T. laniflora leaves was also fractionated by silica gel column chromatography and both the HPLC fingerprint and antimicrobial activity of the obtained fractions were distinct of those originated from the partition process.

Quinine and quinidine are well-known 4-quinolinecarbinolamines that exhibit antimalarial activity, but, in contrast, their epimers 9-epiquinine and 9-epiquinidine are almost inactive. Literature data are conflicting in describing the 4-quinolinecarbinolamine interaction mode with the molecular target, the ferriprotoporphyrin IX [Fe(III)PPIX]. In the present paper, a pharmacophore is proposed based on the binding of the non-aromatic nitrogen to the iron atom. The 4-quinolinecarbinolamine antimalarials were superimposed on the pharmacophore under consideration and complexes with Fe(III)PPIX were constructed. Conformational analyses of the complexes were performed applying the MM+ molecular mechanics method. The analysis of the complexes showed that the proposed ligand mode is possible although it does not explain the activity differences between epimers. A discussion of the structural aspects is also provided.

Four known alkaloids, uleine, epiuleine, apparicine and desmethyluleine, besides the isoprenoids lupeol and stigmasterol were isolated from the stem bark of Aspidosperma parvifolium. Their identification was based on spectroscopic analysis (UV, IR, ¹H-NMR, 13C-NMR, MS). The chromatographic profile of the ethanolic extract was obtained by HPLC and uleine, epiuleine and apparicine were identified in the extract.

Diversas espécies de Ouratea ocorrem no cerrado, sendo utilizadas popularmente como tônico, adstringente e antiinflamatório. No presente trabalho, os extratos hidroetanólicos de O. castanaefolia, O. spectabilis e O. semiserrata foram avaliados para atividade vasodilatadora em anéis de aorta pré-contraídos com fenilefrina. Apenas o extrato de caules de O. semiserrata (OS) produziu vasodilatação significativa (63 ± 3 %, n = 6), na concentração de 100 mg/mL. O fracionamento biomonitorado de OS resultou em uma fração enriquecida em proantocinanidinas, que produziu 100 ± 5 % de vasodilatação (n = 6), na concentração de 100 mg/mL. Os teores totais de proantocianidinas foram determinados para OS e para as frações bioativas, por espectroscopia no visível. Maiores teores de proantocianidinas implicaram aumento da atividade vasodilatadora (r² = 0,9760), sugerindo que compostos desta classe devem ser responsáveis pela atividade do extrato de caules de O. semiserrata.

Ouratea species occur in the cerrado vegetation and some species are popularly used as tonic, astringent and anti-inflammatory. In the present work, we assayed the vasodilator effects of O. castanaefolia, O. spectabilisand O. semiserrata hydroethanolic extracts in aortic rings pre-contracted with phenylephrine. Only O. semiserrata stem extract (OS) produced significant vasodilatation (63 ± 3 %, n = 6), at the concentration of 100 mg/ml. Bioguided fractionation of OS afforded a proanthocyanidin-rich fraction that produced 100 ± 5 % vasodilatation (n = 6; concentration of 100 mg/ml). The total proanthocyanidin contents were assayed for OS and for the bioactive fractions using a spectroscopic method. Higher proanthocyanidin contents implied in increased vasodilator activity (r² = 0.9760), suggesting that compounds of this class may be regarded as responsible for the activity of O. semiserrata stem extract.

Condições otimizadas para a biotransformação de compostos diterpênicos pelo fungo Cephalosporium aphidicola foram determinadas, realizando-se incubações em microescala nas quais variaram-se parâmetros como o tempo de incubação e a quantidade de substrato adicionado. A velocidade (índice) de bioconversão foi monitorada analisando-se os extratos de cultivo através de cromatografia em camada delgada e cromatografia gasosa. Os resultados obtidos mostraram que, para melhores rendimentos, o substrato deve ser adicionado em solução etanólica, na concentração de 15-30 mg por 100 mL de meio de cultura. A extração de produtos deve ser realizada com acetato de etila, a partir da fase aquosa e do micélio, no sexto dia após a adição do substrato. Estes resultados podem ser utilizados como um roteiro para incubações de compostos estruturalmente semelhantes com o fungo Cephalosporium aphidicola.

Parameters for a more efficient biotransformation of diterpene-like compounds by the fungus Cephalosporium aphidicola were established by carrying out microscale feedings at several conditions. Experiments were guided by thin layer chromatography and gas chromatography analysis. It was observed that the substrate should be added in ethanol at concentrations between 15 to 30 mg per 100 ml of medium. The extraction of the product showed to be more efficient when carried out from both mycelia and broth and using ethylacetate as the extracting solvent. The experiment should be stopped six days after feeding the substrate to the fungus for the best product yield.

Four naphthoquinones, lapachol, a-lapachone, dehydro-a-lapachone and 4-hydroxy-a-lapachone were isolated from the ethanol extract of Zeyheria montana M. roots (Bignoniaceae) and were identified by spectroscopic methods. These naphthoquinones, except 4-hydroxy-a-lapachone, were quantified by HPLC, in the crude ethanol extracts of Z. montana roots. The separation of lapachol, a-lapachone and dehydro-a-lapachone was achieved and these compounds were identified in the chromatograms by their retention times and by their on line UV-spectra. The quantification of the three naphthoquinones, in four different samples showed medium contents of 11,0 mg of lapachol, 6,1 mg of a-lapachone and 4,3 mg of dehydro-a-lapachone, in 100 mg of Z.montana roots.

The content of digoxin and lanatoside C (jointly quantified), lanatoside A, lanatoside B, glucoevatromonoside, odorobioside G, glucogitoroside, glucoverodoxine, glucodigifucoside and digitalinum verum was determined by HPLC in Digitalis lanata harvested in Brazil, as well as in clones industrially employed for the production of cardenolides. The Brazilian plants presented greater variation in the contents of cardenolides than the analyzed clones. Lanatoside C and digoxin concentrations were higher in the clones (6120±640 nmol/g dry leaf) than in the Brazilian plants (1820±900 nmol/g dry leaf). The concentrations of these glycosides were found to be within the range described for native species and also were similar to an European cultivar, what makes possible its industrial use for the cardenolides production.

Foi comparada, em laboratório, a atividade moluscicida do extrato hexânico da casca da castanha do caju - Anacardium occidentalel L. (EHCCC), do complexo de cobre (II), docomplexo de chumbo (II) e do ácido anacárdico com objetivo de encontrar entre eles um produto que apresentasse maior estabilidade que o ácido anacárdico. Este foi preparado tratando o EHCCC com hidróxido de chumbo (II) ou com o sulfato de cobre mais hidróxido de sódio ou com hidróxido de cobre (II). Em seguida, o complexo de chumbo (II) ou os complexos de cobre (II)preparados foram tratados com uma solução de ácido sulfúrico diluída. As misturas dos dez produtos obtidos foram testadas sobre caramujos adultos de Biomphalaria glabrata nas concentrações de la 10 ppm. Os mais ativos foram o complexo de cobre (II), obtido com sulfato de cobre mais hidróxido de sódio, e o ácido anacárdico (hidróxido de chumbo) que apresentaram atividade a partir da concentração de 4 ppm. O teor de chumbo do ácido anacárdico (hidróxido de chumbo) foi acima das normas recomendadas pelos Padrões de Saúde Pública dos Estados Unidos.

The molluscicide activity of hexanic extract from Anacardium occidental L. (cashew) nutshell, of copper (II) complex, of lead (II) complex and anacardic acid has been compared in the laboratory in an attempt to obtain better stability than anacardic acid. This was obtained from the hexanic extract of the cashew nut shell by precipitation with lead (II) hydroxide or cupric sulfate plus sodium hydroxide or (II) cupric hydroxide followed by treatment of lead (II) complex with a diluted solution of sulfuric acid. Ten products of the mixture obtained were tested on adults snails of Biomphalaria glabrata at I to 10 ppm. The most active products were copper (II) complex, obtained by cupric sulfate plus sodium hydroxide, and anacardic acid (sodium hydroxide) which presented activity at 4ppm. The anacardic acid's lead content was above the limits accepted by the United States standards.