Avaliação da atividade antiproliferativa dos extratos de Croton urucurana Baillon

O termo câncer refere-se a um conjunto de doenças que atingem e matam milhões de pessoas em todo o mundo. Esta doença é caracterizada pela proliferação descontrolada de células que não respondem mais aos estímulos externos e internos que controlam a proliferação, diferenciação e morte celular. O aumento da expectativa de vida da população é um dos fatores que contribuem para a crescente incidência de câncer no mundo já que o desenvolvimento desta doença está relacionado com o envelhecimento do organismo. O câncer é a segunda maior causa de morte na população mundial e diante de tal índice, fica clara a necessidade de desenvolvimento de novos quimioterápicos. Cerca de 60% das drogas anticâncer são derivadas de produtos naturais, demonstrando assim o grande potencial destes produtos no desenvolvimento de novas drogas anticâncer. Este projeto teve como objetivo avaliar a atividade anticâncer dos extratos brutos e frações obtidas da espécie vegetal Croton urucurana Bailon. Os extratos brutos diclorometânico (EBD) e etanólico (EBE) foram avaliados em ensaio de citotoxicidade in vitro em cultura de células tumorais humanas. Os dois extratos apresentaram atividade in vitro, porém somente o EBD foi submetido ao processo cromatográfico de fracionamento, sendo as frações obtidas deste processo submetidas ao teste antiproliferativo in vitro. O EBD também foi utilizado para avaliar sua atividade em modelo de tumor sólido de Ehrlich. Neste teste, esse extrato apresentou inibição do crescimento tumoral apenas com a menor dose. Como o EBE também apresentou atividade antiproliferativa in vitro, é importante dar continuidade aos estudos com este extrato, com o objetivo de avaliar sua atividade em modelos experimentais in viv

Anticancer and antiestrogenic activities of extracts and fractions of leaves of "Psidium guajava" L.

Orientadores: João Ernesto de Carvalho, Ana Lúcia Tasca Gois Ruiz, Mary Ann FoglioTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: Atualmente cerca de 75% das drogas anticâncer são derivadas de produtos naturais, demonstrando o grande potencial destes produtos na pesquisa e desenvolvimento de novos fármacos para o tratamento desse conjunto de doenças. Estudos anteriores, realizados no Centro Pluridisciplinar de Pesquisas Químicas, Biológicas e Agrícolas (CPQBA - Unicamp), revelaram que os extratos e os princípios ativos obtidos da "Psidium guajava" L. apresentam potencial ação anticâncer tanto em cultura de células tumorais humanas quanto em modelos experimentais com animais de laboratório. O tratamento com a fração enriquecida com os princípios ativos (Guajadial e Psidial A) da espécie, por via intraperitoneal, resultou na inibição do crescimento dos tumores sólidos de Ehrlich em todas as doses utilizadas (10, 30 e 50 mg/kg). O aumento significativo do peso e do tamanho dos úteros dos animais tratados com a fração sugeriu uma possível ação hormonal dos princípios ativos, que foi então avaliada utilizando a técnica de docking molecular in silico. Tanto o Guajadial quanto o Psidial A apresentaram afinidade significativa a ambas as isoformas dos receptores de estrógeno (ER-"alfa" e ER-"beta"), indicando, portanto, possível ação sobre os receptores hormonais. Neste contexto, este trabalho avaliou, tanto in vitro quanto in vivo, a atividade antiproliferativa e ação estrogênica/antiestrogênica do extrato bruto e frações obtidos a partir das folhas de "P. guajava" L.. Por meio de diversas técnicas cromatográficas, foram obtidas frações ativas das quais, as frações FB1c e FFINAL apresentaram atividade antiproliferativa in vitro, com seletividade para as linhagens tumorais humanas MCF-7 e MCF-7 BUS (TGI = 5,59 e 2,27 µg/mL, respectivamente), sugerindo desta forma, uma possível correlação entre a atividade antiproliferativa e ação hormonal. Esta correlação foi confirmada tanto nos testes de E-screen quanto nos testes in vivo que avaliam atividade uterotrófica, nos quais a fração FFINAL foi capaz de bloquear o efeito proliferativo do estradiol nas células MCF-7 BUS e no útero de ratas pré-púberes. Os modelos in vivo de tumor sólido de Ehrlich e Hollow fiber, empregando-se a via oral como rota de tratamento, confirmaram a atividade anticâncer das frações de P. guajava evidenciada através da atividade antiproliferativa in vitro. Os testes que avaliam os mecanismos de morte celular, por meio da citometria de fluxo, revelaram que as frações FB1c (5 e 10 µg/mL) e a FFINAL (2,5 µg/mL), após 24 horas de tratamento, foram capazes de induzir a externalização dos resíduos de fosfatidilserina nas membranas citoplasmáticas das células MCF-7 e MCF-7 BUS, sugerindo desta forma, uma morte por meio do processo de apoptose. Comparativamente à ação do tamoxifeno sobre o ciclo celular, o tratamento com a fração FFINAL sobre as células MCF-7 BUS provocou a parada do ciclo celular na fase G1, em todos os tempos avaliados (24, 30 e 48 horas). Por fim, não foi possível evidenciar o efeito anticâncer do extrato das folhas de P. guajava no modelo de carcinogênese induzida quimicamente pelo composto 1- metil,1-nitrosoureia (MNU) porque este modelo apresentou uma baixa taxa de indução de tumores de mama após 120 dias da administração do carcinógenoAbstract: Currently around 75% of anticancer drugs are derived from natural products, showing the great potential of these products in new drug research and development for the treatment of this set of diseases. Previous studies carried out at the Center for Chemical, Biological and Agricultural Research (CPQBA - Unicamp) revealed that extracts and active ingredients obtained from "Psidium guajava" L. show potential anticancer action both in human tumor cell culture and in experimental models with laboratory animals. Intraperitoneal treatment with the enriched fraction (Guajadial and Psidial A) of the species resulted in inhibition of the growth of Ehrlich solid tumors at all doses (10, 30 and 50 mg/kg). The significant increase in the weight and size of the uteri of the animals treated with the fraction suggested a possible hormonal action of the active principles, which was then evaluated using the in silico molecular docking technique. Both Guajadial and Psidial A showed significant affinity for both isoforms of estrogen receptors (ER-"alfa" and ER-"beta"), thus indicating a possible action on hormone receptors. In this context, this work evaluated, both in vitro and in vivo, the antiproliferative activity and estrogenic/antiestrogenic action of the crude extract and fractions obtained from "P. guajava" L. leaves. By chromatographic techniques fractions were obtained, some active ones including FB1c and FFINAL that showed antiproliferative activity in vitro with selectivity for human tumor cell lines MCF-7 and MCF-7 BUS (TGI = 5, 59 and 2.27 mg / mL, respectively), thus demonstrating a possible correlation between the antiproliferative activity and hormone action. This correlation was confirmed in both the E-screen tests for in vivo tests that assess uterotrophic activity, which found that the FFINAL was able to block the proliferative effect of estradiol on the MCF-7 cells BUS and uterus rats pre-pubescent. The anticancer activity was confirmed by in vivo Ehrlich's solid tumor model and by Hollow fiber assay, after oral treatment. The tests that evaluate the mechanisms of cell death, by flow cytometry showed that fractions FB1c (5 and 10 ug / ml) and FFINAL (2.5 ug / ml) after 24 hours of treatment, were able to induce externalization of phosphatidylserine residues in the cytoplasmic membranes of MCF-7 and MCF-7 cells BUS, suggesting, therefore, death by apoptosis. Compared to the action of tamoxifen on the cell cycle, treatment with FFINAL of MCF-7 BUS cells caused cell cycle arrest in the G1 phase of the cycle, at all time periods evaluated (24, 30 and 48 hours). Finally, the test chemical carcinogenesis induced by compound 1-methyl-1-nitrosourea (MNU), demonstrated that the induction of breast tumors was not effective with low index tumors 120 days after carcinogen administrationDoutoradoFarmacologia, Anestesiologia e TerapeuticaDoutora em Odontologia2014/01283-9FAPES

Liquid-liquid Equilibrium Data For Fatty Systems Containing Monoacylglycerols And Diacylglycerols

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Edible oils can be deacidified by solvent extraction, and the comprehension of liquid liquid equilibrium is essential to the design process. Thus, this study aims to determine the data of liquid liquid equilibrium for the fatty systems containing triacylglycerols (TAG) of refined oils (soybean, cottonseed, and rice bran) + commercial fatty acid (linoleic and oleic) + diacylglycerols (DAG) of commercial mixture of mono- and diacylglycerols + monoacylglycerols (MAG) of commercial mixture of mono- and diacylglycerols + anhydrous ethanol at different temperatures (303.15 K and 318.15 K). The high performance size exclusion chromatography (HPSEC) method was used for quantification of all components: TAG, DAG, MAG, fatty acids, and ethanol in both phases in equilibrium. The preference of fatty acids to solvent phase is consistent to previously reported data in literature. The mass fraction of MAG was more than 2 times greater in the solvent phase, and the DAG concentration is higher in the oil phase. The average deviation between experimental and calculated compositions, using NRTL model, was less than 1 %. The obtained parameter set enables the simulation of liquid liquid extractors for edible soybean, cottonseed, considering the complex multicomponent mixtures' composed of tri-, di-, and monoacylglycerols and rice bran oil deacidification, and fatty acids.60823712379Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)CAPES [1089097]FAPESP [2008/56258-8]CNPq [483340/2012-0, 406856/2013-3, 305870/2014-9, 309780/2014-4

Anti-Estrogenic Activity of Guajadial Fraction, from Guava Leaves (Psidium guajava L.)

The research of natural products has allowed for the discovery of biologically relevant compounds inspired by plant secondary metabolites, which contributes to the development of many chemotherapeutic drugs used in cancer treatment. Psidium guajava leaves present a diverse phytochemical composition including flavonoids, phenolics, meroterpenoids, and triterpenes as the major bioactive constituents. Guajadial, a caryophyllene-based meroterpenoid, has been studied for potential anticancer effects tested in tumor cells and animal experimental models. Moreover, guajadial has been reported to have a mechanism of action similar to tamoxifen, suggesting this compound as a promisor phytoestrogen-based therapeutic agent. Herein, the anti-estrogenic action and anti-proliferative activity of guajadial is reported. The enriched guajadial fraction was obtained by sequential chromatographic techniques from the crude P. guajava dichloromethane extract showing promising anti-proliferative activity in vitro with selectivity for human breast cancer cell lines MCF-7 and MCF-7 BUS (Total Growth Inhibition = 5.59 and 2.27 µg·mL−1, respectively). Furthermore, evaluation of anti-estrogenic activity in vivo was performed demonstrating that guajadial enriched fraction inhibited the proliferative effect of estradiol on the uterus of pre-pubescent rats. These results suggest a relationship between anti-proliferative and anti-estrogenic activity of guajadial, which possibly acts in tumor inhibition through estrogen receptors due to the compounds structural similarity to tamoxifen

Characterization Of A Refinement Of The Pylorus Ligation" Model Of Rat Gastric Ulceration Resulting In "no Pain" And A More Specific Pharmacological Response."

The pharmacological assessment of the factors for gastric protection of a test substance should involve experimental models that can determine the involvement of cytoprotective factors, as well as their influence on the secretion of hydrochloric acid. The original protocol of pylorus ligation in rats proposed by Shay et al. in 1945, still in use today, provides a latency time of 240 min without considering the effect of postoperative pain in the mechanisms of peptic ulcer. This paper proposes a modification of this experimental protocol by eliminating the pain throughout the postoperative period, as a refinement of the test with consequent improvement of the pharmacological response. Adult male Wistar/Uni rats underwent surgical ligation of the pylorus and were kept anesthetized throughout the experimental period (4h) in contrast to the other experimental groups that followed the original protocol proposed by Shay et al., 1945. We were able to determine effective doses for a positive control, as well as of a variety of secretagogues in the new experimental protocol proposed. The suppression of post-surgical pain, through the use of anesthesia throughout the experimental period, brought several benefits for the study of gastric acid secretion, rendering a more homogeneous pharmacologic response in non-inbred animals, thus being an effective experimental procedure.67121-

Lupeol and its esters: NMR, powder XRD data and in vitro evaluation of cancer cell growth

<div><p>ABSTRACT The triterpene lupeol (1) and some of its esters are secondary metabolites produced by species of Celastraceae family, which have being associated with cytotoxic activity. We report herein the isolation of 1, the semi-synthesis of eight lupeol esters and the evaluation of their in vitro activity against nine strains of cancer cells. The reaction of carboxylic acids with 1 and DIC/DMAP was used to obtain lupeol stearate (2), lupeol palmitate (3) lupeol miristate (4), and the new esters lupeol laurate (5), lupeol caprate (6), lupeol caprilate (7), lupeol caproate (8) and lupeol 3’,4’-dimethoxybenzoate (9), with high yields. Compounds 1-9 were identified using FT-IR, 1H, 13C-NMR, CHN analysis and XRD data and were tested in vitro for proliferation of human cancer cell activity. In these assays, lupeol was inactive (GI50> 250µg/mL) while lupeol esters 2 -4 and 7 - 9 showed a cytostatic effect. The XRD method was a suitable tool to determine the structure of lupeol and its esters in solid state. Compound 3 showed a selective growth inhibition effect on erythromyeloblastoid leukemia (K-562) cells in a concentration-dependent way. Lupeol esters 4 and 9 showed a selective cytostatic effect with low GI50 values representing promising prototypes for the development of new anticancer drugs.</p></div

X-ray structure of O-methyl-acrocol and anti-cancer, anti-parasitic, anti-bacterial and anti-Zika virus evaluations of the Brazilian palm tree Acrocomia totai

Acrocomia totai Mart ("macauba") is a palm tree native from Brazil, whose potential for biodiesel production has been widely explored. In spite of the industrial interest in the oil from the nuts, little is known about the potential applications of other parts of the plant, especially in the pharmacological area. A phytochemical study of the plant thorns led to the identification of a new compound 3-(R)-methoxy-21-(R)-H-hop-22(29)-en-30-ol 1, two known triterpenes 2-3, four steroids 4-7 and a stilbene, piceatannol 8. The structures were elucidated by spectroscopic analyses, including 1D and 2D NMR and low and high resolution mass spectrometry. Compound 1 was purified as crystals, which allowed the determination of the absolute configuration of the asymmetric carbons by analysis of the X-ray diffraction spectrum. Biological tests were performed with crude extract (CE), fractions and isolated compound. The assays showed activity for CE against lung carcinoma (GI(50) 59.2 mu g mL(-1)). The ethyl acetate fraction (EAF) showed efficacy against many tumor cell lines, and the tests showed the most prominent activity for breast cancer (GI(50) 10.4 mu g mL(-1)), glioma (GI(50) 77.3 mu g mL(-1)), uterine cervix (SiHa) HPV 16 (IC50 39.8 mu g mL(-1)), (HeLa) HPV 18 (IC50 12.0 pg mL(-1)) and Caco-2 (IC50 40.0 mu g mL(-1)) and showed bacteriostatic action against Staphylococcus aureus (MIC 50 mu g mL(-1)). Piceatannol 8 isolated from EAF showed activity against the protozoan which causes leishmaniosis (IC50 58.4 mu g mL(-1)). For Ttypanosorna cruzi, the methanol fraction (EC50 15.5 mu g mL(-1)), CE (20.5 mu g(-1)), and HEF (43.8 mu g mL(-1)) were the most active, being highly selective for the protozoan and less toxic against Vero cells. The compound 8 was further tested against Zika virus MR 766 strain on MOI 2, however the assays showed no inhibition against virus infection109483492CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO ARAUCÁRIA DE APOIO AO DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO DO ESTADO DO PARANÁ - FAFUNDAÇÃO DE AMPARO À PESQUISA E INOVAÇÃO DO ESTADO DE SANTA CATARINA - FAPESCsem informaçãosem informaçãosem informaçãosem informaçã