Caio Pinho Fernandes

A new tropane alkaloid from the leaves of Erythroxylum subsessile isolated by pH-zone-refining counter-current chromatography.

Tropane alkaloids are bioactive metabolites with great importance in the pharmaceutical industry and the most important class of natural products found in the Erythroxylum genus. However, these compounds are usually separated by traditional chromatographic techniques, in which the sample is progressively purified in multiple chromatographic steps, resulting in a time- and solvent-consuming procedure. In this work we present the isolation of a novel alkaloid, 6β,7β-dibenzoyloxytropan-3α-ol, together with the two known 3α-benzoyloxynortropan-6β-ol and 3α,6β-dibenzoyloxytropane alkaloids, directly from the crude alkaloid fraction from the leaves of Erythroxylum subsessile, by using a single run pH-zone-refining counter-current chromatography method. The ethyl acetate/water (1:1, v/v) biphasic solvent system with triethylamine and HCl as retention and eluter agents, respectively, was used to isolate tropane alkaloids for the first time. The structures of the isolated alkaloids were elucidated by spectroscopic methods.

Study of the non-clinical healing activities of the extract and gel of Portulaca pilosa L. in skin wounds in wistar rats: A preliminary study

INTRODUCTION Portulaca pilosa L., belonging to the family Portulacaceae, is a common herbaceous plant in the Americas and in the Amazon, is popularly known as love-grown and is traditionally used as an aid in the treatment of burns, buds, insect bites and wound healing. This study aims to evaluate the non-clinical topical healing activity of the P. pilosa gel (GPP) and the propyleneglycol extract of P. pilosa (EPP) in Wistar rats. METHODS For the healing activity, wistar rats were divided into the following groups: negative control (GVE - vehicle, 150mg/kg), positive control (Fibrinase®- FIB, 100U/kg), Portulaca pilosa gel 10%, (GPP, 150mg/kg), and propylenglycollic extract of Portulaca pilosa (EPP, 150 mg/kg), which were submitted to a surgical procedure to obtain the wounds, and were treated topically for 7days. After treatment, the treated area was removed and a histopathological analysis was performed. RESULTS The EPP when analyzed in HPLC was able to identify the presence of gallic acid. EPP significantly modulated the tissue inflammatory response, presenting low number of inflammatory cells in the histopathological study. Treatment with EPP and GPP significantly stimulated angiogenesis and this response was superior to the fibrinase® group. Treatment with EPP and GPP significantly stimulated the proliferation of fibroblasts. The groups treated with EPP and GPP presented an organization pattern of the epidermis and dermis better than the control group, with a mild inflammatory process, with fibroblast proliferation and increased formation of collagen fibers. CONCLUSION Thus, from the results obtained it can be suggested that the phytochemical marker of the P. pilosa species for healing activity is gallic acid and, together with the macroscopic and microscopic findings triggered by the topical applications of EPP and GPP, it can be concluded that this plant species has topical healing activity, with great potential for use, since this pharmacological action is associated with a possible topical anti-inflammatory activity.

Aniseed (Pimpinella anisum, Apiaceae) Oils

Abstract Pimpinella anisum, commonly known as aniseed, is one of the oldest species used by people, being cultivated in Egypt and later in Greece, Rome, and the Middle East. Anise has white flowers and yellow-brown or green-brown fruits, which contain not less than 2% (w/w) of essential oil. Egypt and Spain are the worlds biggest producers of this essential oil. Its fruits and essential oil are widely used in the food industry as a flavoring, antioxidant, antispoilage agent, and preservative in many products, such as candies, sweets, toffees, and beverages. Many countries around the Mediterranean region, such as Turkey, Greece, Italy, Spain, and France, have traditional alcoholic beverages produced with P. anisum, such as anis, arak, pastis, ouzo, sambuca, zivania, and raki. This chapter provides relevant information regarding this species in the food industry context.

Ethnopharmacological Studies for the Development of New Drugs

1Laboratório de Pesquisa em Fármacos, Departamento de Ciências Biológicas e da Saúde, Universidade Federal do Amapá, Macapá, AP, Brazil 2Laboratório de Nanobiotecnologia Fitofarmacêutica, Universidade Federal do Amapá, Macapá, AP, Brazil 3Facultad de Farmácia, Universidad de Oriente, Santiago de Cuba, Cuba 4Laboratório de Farmacologia de Productos Naturales, Departamento de Farmácia, Facultad de Quı́mica, Universidad Nacional Autónoma de México, Ciudad de México, Mexico 5Laboratório de Neurociências e Ensaios Farmacológicos, Departamento de Fisiologia, Centro de Ciências Biológicas e da Saúde, Universidade Federal de Sergipe, São Cristóvão, SE, Brazil

Chemical Constituents of Essential Oils from Leaves of Two Erythroxylum Species

Erythroxylum P. Browne is the most representative genus from the Erythroxylaceae family, being constituted by approximately 97% of its species [1]. They are mainly spread through tropical and subtropical regions such as South America, Africa, and Madagascar [2–4]. Brazil has some species spread through “restinga,” a typical sandy coastal plain originated from quaternary marine deposits [5, 6]. This genus is recognized as a tropane alkaloid producer [7] and also by the presence of terpenoids and flavonoids [8, 9]. However, few studies were performed concerning the volatile constituents of this genus [10]. Erythroxylum ovalifolium Peyr. and Erythroxylum subsessile (Mart.) O. E. Schulz are shrubs that grow spontaneously at Restinga de Jurubatiba National Park (Rio de Janeiro, Brazil). They are commonly known at this location as “quixaba” and used in folk medicine to treat backache [6]. However, to our knowledge, both species remain unexplored regarding their chemical constituents. In this context, the aim of the present study was to describe for the first time the chemical composition of essential oils obtained from leaves of these two Erythroxylum species. Leaves of E. ovalifolium (S22 14.722 , W41 34.916 ) and E. subsessile (S22 16.111 , W41 38.990 ) were collected in January 2009 at Restinga de Jurubatiba National Park, Rio de Janeiro, Brazil. Identification of the plants was carried out by Dr. Marcelo G. Santos. Voucher samples were deposited at the Herbarium of the Rio de Janeiro State University, Sao Goncalo, under the register numbers RFFP 2147 and RFFP 2126, respectively. Fresh leaves (400 g per plant) of E. ovalifolium and E. subsessile were individually ground with distilled water and subjected to hydrodistillation using a Clevenger-type apparatus for 4 h. Essential oils were dried over anhydrous sodium sulfate, and the yields were 0.04% (w/w) and 0.05% (w/w) based on fresh leaves of E. subsessile and E. ovalifolium, respectively. The low yields observed for these essential oils are in accordance with literature data for the genus Erythroxylum [7]. The gas chromatographic (GC) conditions were as follows: injector temperature 260 C, detector temperature 280 C, carrier gas helium, flow rate 1 mL/min, and split injection with split ratio 1:40. Oven temperature was initially 60 C and then raised to 240 C at a rate of 3 C min–1. Each essential oil was dissolved in CH2Cl2 (1:100 mg/ L) for the analysis. Gas-chromatographic analysis revealed a total of 20 constituents in E. ovalifolium essential oil, corresponding to 91.3% total relative composition of this oil, and 22 constituents in E. subsessile essential oil, corresponding to 99.7% of total relative composition of this oil. (2E)-Hexenal (24.1%) and (E)-ocimene (23.2%) were found to be the major compounds of E. ovalifolium essential oil, while (2E)-hexenal (41.0%) and -pinene (20.0%) were found to be the major compounds of E. subsessile essential oil. All identified substances and their relative percentage are presented in Table 1.

Molluscicidal activity of Manilkara subsericea (Mart.) dubard on Biomphalaria glabrata (Say, 1818)

Schistosomiasis is promoted for species from Schistosoma genus affecting over 200 million people worldwide. Molluscicides are an efficient method to control this disease, being able to reduce intermediate host snail Biomphalaria glabrata number. In function of resistance cases using niclosamide, natural products are promisors to discover new drugs. Manilkara subsericea is endemic to Brazilian sandbanks of Rio de Janeiro State and wide ranges of biological activities. However, there is no studies evaluating its effects as molluscicidal agent. We tested crude extract from leaves of M. subsericea molluscicidal action, as well it ethyl-acetate fraction and isolated substances against B. glabrata. M. subsericea leaves crude extract and ethyl acetate fraction induced 80±4.13% and 86.66±4.59% mortality of adult snails at concentrations of 250ppm after 96h, and their LD50 values were 118.7±1.62 and 23.41±1.15ppm respectively. Isolated substances from M. subsericea were also considered active. Quercetin, myricetin and ursolic acid, at concentration of 100ppm (96h), were able to induce mortality levels of 100%, 80% and 53.33%, respectively. Our results suggest that M. subsericea can be considered promising as a molluscicide agent.

Plants of Brazilian restingas with tripanocide activity against Trypanosoma cruzi strains

Chagas disease is caused by the Trypanosoma cruzi affecting millions of people, and widespread throughout Latin America. This disease exhibits a problematic chemotherapy. Benznidazole, which is the drug currently used as standard treatment, lamentably evokes several adverse reactions. Among other options, natural products have been tested to discover a novel therapeutic drug for this disease. A lot of plants from the Brazilian flora did not contain studies about their biological effects. Restinga de Jurubatiba from Brazil is a sandbank ecosystem poorly studied in relation to plant biological activity. Thus, three plant species from Restinga de Jurubatiba were tested against in vitro antiprotozoal activity. Among six extracts obtained from leaves and stem parts and 2 essential oils derived from leave parts, only 3 extracts inhibited epimastigote proliferation. Substances present in the extracts with activity were isolated (quercetin, myricetin, and ursolic acid), and evaluated in relation to antiprotozoal activity against epimastigote Y and Dm28 Trypanosoma cruzi strains. All isolated substances were effective to reduce protozoal proliferation. Essentially, quercetin and myricetin did not cause mammalian cell toxicity. In summary, myricetin and quercetin molecule can be used as a scaffold to develop new effective drugs against Chagas’s disease.

Non-clinical toxicity study of the oil-resin and vaginal cream of Copaiba (Copaifera duckei, Dwyer)

The toxic effects of copaiba oil-resin (ORC) and the copaiba oil-resin vaginal cream (CVC) were evaluated in subacute treatment phase, in wistar rats, which were treated orally (p.o) and intravagin...