Actividad in vitro anti-Leishmania (Trypanosomatidae) del epóxido trans-Z-&-bisaboleno y del Safrol, en frutos de Piper auritum (Piperaceae) / Anti-Leishmania (Trypanosomatidae) activity of trans-Z-&-bisabolene epoxide and isolation of safrole, in fruits of Piper auritum (Piperaceae)

Resumen La leishmaniasis es una enfermedad con una alta incidencia en el ser humano, que puede ser controlada, pero como los tratamientos tienen efectos secundarios importantes se han realizado estudios de diversas plantas con el fin de encontrar compuestos con actividad antileishmaniásica que presenten pocos efectos nocivos para el ser humano. El presente estudio consistió en realizar un tamizaje fitoquímico de la planta, para identificar la presencia de cumarinas, terpenos, triterpenos y azúcares reductores. El objetivo fue encontrar componentes químicos puros con actividad contra el parásito Leishmania sp. Por tal razón se purificaron los compuestos: trans-Z-alfabisaboleno y el Safrol, a los que se les realizaron pruebas del efecto anti parasitario que presentaron un CI50 de 50.0 µg/mL y 0.0 µg/mL, respectivamente. Además, se discute la importancia de estos nuevos hallazgos. El compuesto mayoritario presente en los aceites esenciales (Safrol) no es el componente que presentó la actividad. Es importante realizar estudios sobre su proyección en el tratamiento de la leishmaniasis.

Abstract The leishmaniosis disease incidence is high in tropical regions, and its current treatment has shown severe secondary effects. Considering this problem, many studies have focused on plants, looking for chemical components that have anti-leishmanial activity, and are free of adverse effects for human beings. The purpose of this work was to find a chemical component with this kind of activity in Piper auritum. In a phytochemical screening of this plant, we found some cumarins, terpens, triterpens and reducing sugars; and later, we identified the components trans-Z-α-bisabolene epoxide and Safrol. The first component presented a CI50 of 50.0 µg/mL of anti-Leishmania activity. The Safrol, which is the major component of the essential oils of this plant, did not show antiparasitic activity. These results are discussed considering treatment of leishmaniasis. Rev. Biol. Trop. 66(2): 826-835. Epub 2018 June 01.

Metabolism and antifungal activity of safrole, dillapiole, and derivatives against botryodiplodia theobromae and colletotrichum acutatum / Metabolismo y actividad antifúngica de safrol, dilapiol, y derivados contra botryodiplodia theobromae y colletotrichum acutatum

The direct in vitro fungitoxicity and metabolism of safrole and dillapiole (isolated from Piper auritum and Piper holtonii, respectively) by Botryodiplodia theobromae and Colletotrichum acutatum were investigated. Higher values of mycelial growth inhibition for both fungi were obtained for dillapiole, as compared with safrole. B. theobromae was able to metabolize both compounds to their respective vicinal diols, reaching 65 percent relative abundance during the biotransformation of dillapiole; while C. acutatum only transformed safrole to various metabolites with relative abundances under 5 percent. According to the low antifungal activity of the major metabolic products (< 5 percent for vicinal diols), a detoxification process was implied. Studies on the influence of some substituents in the aromatic ring of safrole and dillapiole on the antifungal activity against B. theobromae were also carried out. As result, the safrole nitrated derivative, 6-nitrosafrole, showed a fungitoxicity level similar to that displayed by the commercial fungicide Carbendazim® under the conditions used. In light of this, safrole and dillapiole could be suggested as feasible structural templates for developing new antifungal agents.

Se investigó la fungitoxicidad directa in vitro y el metabolismo de safrol y dilapiol (obtenidos desde Piper auritum and Piper holtonii, respectivamente) por Botryodiplodia theobromae y Colletotrichum acutatum. Los valores mayores de inhibición del crecimiento micelial de ambos hongos se obtuvieron para dilapiol, en comparación con safrol. B. theobromae metabolizó ambos compuestos a sus respectivos dioles vecinales, alcanzando abundancias relativas del 65 por ciento durante la biotransformación del dilapiol; mientras que C. acutatum solo transformó safrol en varios metabolitos con abundancias relativas menores al 5 por ciento. De acuerdo con la baja actividad antifúngica de los productos metabólicos mayoritarios (< 5 por ciento para los dioles vecinales), se sugiere un proceso de desintoxicación. Adicionalmente, se evaluó la influencia de algunos sustituyentes en el anillo aromático de safrol y dilapiol sobre la actividad antifúngica contra B. theobromae. Como resultado, el derivado nitrado del safrol, el 6–nitro safrol, presentó un nivel de fungitoxicidad similar al exhibido por el fungicida comercial Carbendazim® bajo las condiciones usadas. A la luz de lo anterior, safrol y dilapiol podrían ser sugeridos como plantillas estructurales adecuadas para el desarrollo de nuevos agentes antifúngicos.

Yield of essential oil and safrole content based on fresh and dry biomass of long pepper in the Brazilian Amazon / Rendimento do óleo essencial e teor de safrol com base em biomassa fresca e seca de pimenta longa na Amazônia brasileira

Long pepper (Piper hispidinervum) is an Amazonian species of commercial interest due to the production of safrole. Drying long pepper biomass to extract safrole is a time consuming and costly process that can also result in the contamination of the material by microorganisms. The objective of this study was to analyze the yield of essential oil and safrole content of fresh and dried biomass of long pepper accessions maintained in the Active Germoplasm Bank of Embrapa Acre, in the state of Acre, Brazil, aiming at selecting genotypes with best performance on fresh biomass to recommend to the breeding program of the species. Yield of essential oil and safrole content were assessed in 15 long pepper accessions. The essential oil extraction was performed by hydrodistillation and analyzed by gas chromatography. A joint analysis of experiments was performed and the means of essential oil yield and safrole content for each biomass were compared by Student's t-test. There was variability in the essential oil yield and safrole content. There was no difference between the types of biomass for oil yield; however to the safrole content there was difference. Populations 9, 10, 12 and 15 had values of oil yield between 4.1 and 5.3%, and safrole content between 87.2 and 94.3%. The drying process does not interfere in oil productivity. These populations have potential for selection to the long pepper breeding program using oil extraction in the fresh biomass.

A pimenta longa (Piper hispidinervum) é uma espécie amazônica de interesse comercial devido à produção de safrol. A secagem da biomassa para extração do óleo essencial demanda tempo e acarreta custos, podendo também resultar em contaminação do material por microorganismos. O objetivo deste estudo foi analisar o rendimento de óleo essencial e teor de safrol da biomassa verde e seca de acessos de pimenta longa mantida no Banco Ativo de Germoplasma da Embrapa Acre a fim de selecionar genótipos com melhor desempenho em biomassa verde para recomendação ao programa de melhoramento genético da espécie. O rendimento do óleo essencial e o teor de safrol foram avaliados em 15 acessos de pimenta longa. Foi realizada uma análise conjunta de experimentos e as médias do rendimento do óleo essencial e do teor de safrol para cada população foram comparadas pelo teste t de Student. Houve variabilidade do rendimento de óleo essencial e do teor de safrol. Não houve diferença entre os tipos de biomassa para o rendimento do óleo, já para o teor de safrol houve diferença. As populações 9, 10, 12 e 15 mostraram valores de rendimento de óleo entre 4,1 e 5,3%, e teor de safrol entre 87,2 e 94,3%. O processo de secagem não interfere na produtividade de óleo. Estas populações têm potencial para seleção para o programa de melhoramento genético de pimenta longa por meio da extração de óleo essencial da biomassa verde.

Chemical composition and bioactivity of Piper auritum and P. multiplinervium essential oils against the red flour beetle, Tribolium castaneum (Herbst) / Composición química y bioactividad de los aceites esenciales de Piper auritum y P. multiplinervium contra el escarabajo rojo de la harina, Tribolium castaneum (Herbst)

Stored grain insects have shown resistance to synthetic insecticides, fact that has promoted the use of vegetable species for integrated pest management. Piper auritum and P. multiplinervium are two plants from the Piperaceae family present in the department of Chocó, Colombia, one of the most important hot spots of biodiversity in the world. This study was conducted to determine the repellent activity and toxicity of essential oils (EOs) isolated from these plants against Tribolium castaneum, using the area preference and contact toxicity methods, respectively. P. auritum EO presented greater repellency than P. multiplinervium, the first showed 100 percent lethality at minimum tested exposure period (24 h) whereas the second reached 16 percent at 72 h. EOs were analyzed by gas chromatography-mass spectrometry. P. auritum major components were safrole (93.2 percent) and miristicine (4.3 percent), whereas for P. multiplinervium were beta-elemene (9.0 percent), trans-beta-caryophyllene (5.3 percent) and caryophyllene oxide (4.1 percent). It is speculated that the repellent effect of P. auritum may be related to its safrole content, a known repellent. These results evidenced Piper species could be used for development of repellents against T. castaneum.

Los insectos de los granos almacenados han mostrado resistencia a los insecticidas sintéticos, hecho que ha promovido el uso de especies vegetales para el manejo integrado de plagas. Piper auritum y P. multiplinervium son dos plantas de la familia Piperaceae presentes en el departamento del Chocó, Colombia, uno de los puntos de biodiversidad más importantes del mundo. En este estudio fue determinada la actividad repelente y toxicidad de los aceites esenciales (AE) aislados de estas plantas contra Tribolium castaneum, utilizando el método de área de preferencia y toxicidad por contacto, respectivamente. El AE de P. auritum presentó mayor repelencia que el de P. multiplinervium, el primero mostró 100 percent de letalidad al menor tiempo de exposición (24 h), mientras que el segundo alcanzó el 16 percent a las 72 h. Los AEs fueron analizados por cromatografía de gases-espectrometría de masas. Los componentes principales de P. auritum fueron safrol (93.2 percent) y miristicina (4.3 percent), mientras que para P. multiplinervium fueron beta-elemene (9.0 percent), trans-beta-cariofileno (5.3 percent) y óxido de cariofileno (4.1 percent). Se cree que el efecto repelente de P. auritum puede estar relacionado con su contenido de safrol, un repelente conocido. Estos resultados evidencian que las especies de Piper podrían ser utilizadas para el desarrollo de repelentes contra T. castaneum.

alpha-Tocopheryl Succinate Inhibits Osteoclast Formation by Suppressing Receptor Activator of Nuclear Factor-kappaB Ligand (RANKL) Expression and Bone Resorption

OBJECTIVE: Osteoclasts are bone-resorbing multinucleated cells derived from the monocyte/macrophage lineage during normal and pathological bone turnover. Recently, several studies revealed that alpha-tocopheryl succinate (alphaTP-suc) have demonstrated potent anti-cancer activities in vitro and in vivo. However, the effects of alphaTP-suc on osteoclast formation and bone resorption remain unknown. Thus, in this study, we examined the effects of alphaTP-suc on osteoclast differentiation and bone resorbing activity in inflammatory bone loss model. METHODS: Osteoclast differentiation assay was performed by cocultures of mouse bone marrow cells and calvarial osteoblasts in culture media including interleukin-1 (IL-1). Osteoclasts were stained for tartrate-resistant acid phosphatase (TRAP). The level of receptor activator of nuclear factor-kappaB ligand (RANKL) mRNA was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). ICR mice were administered an intraperitoneal injections of alphaTP-suc or dimethyl sulfoxide (DMSO) 1 day before the implantation of a freeze-dried collagen sponge loaded with phosphate-buffered saline (PBS) or IL-1 over the calvariae and every other day for 7 days. The whole calvariae were obtained and analyzed by micro-computed tomography (CT) scanning, and stained for TRAP. RESULTS: alphaTP-suc inhibits osteoclast formation in cocultures stimulated by IL-1 and decreased the level of expression of RANKL mRNA in osteoblasts. In addition, administered intraperitoneal injections of alphaTP-suc prevented IL-1-mediated osteoclast formation and bone loss in vivo. CONCLUSION: Our findings suggest that alphaTP-suc may have therapeutic value for treating and preventing bone-resorptive diseases, such as osteoporosis.

Estudo da adaptação da espécie Piper hispidinervum C. DC. (pimenta longa) à região do Vale do Itajaí - SC, através da composição química do óleo essencial obtido por hidrodestilação por micro-ondas e convencional / Adaptation studies of Piper hispidinervum C. DC. (long pepper) species in Itajaí Valley - SC, by the chemical composition of essential oil obtained by microwave and traditional hydrodistillation

Neste trabalho, a espécie Piper hispidinervum (pimenta longa), endêmica do estado do Acre, foi cultivada na região do Vale do Itajaí - SC, e sua adaptação foi avaliada em relação à composição química do óleo essencial obtido pelos processos de hidrodestilação convencional e por micro-ondas. O safrol foi identificado como o constituinte majoritário do óleo essencial desta espécie, o qual foi empregado como parâmetro de avaliação da capacidade de adaptação desta planta à região sul do Brasil, já que a proposta abrange avaliar a utilização desta espécie como fonte alternativa de safrol, em substituição a Canela Sassafrás (Ocotea odorifera), espécie muito explorada até a década de 90 nesta região. As amostras da planta foram obtidas de diferentes regiões do Acre e foram cultivadas na estação experimental da EPAGRI - Itajaí-SC. O óleo essencial das folhas forneceu um teor médio de safrol entre 76,6% e 89,9%. A análise por CG-DIC e CG-EM do óleo com maior concentração de safrol, apresentou os seguintes constituintes: safrol (89,93%), α-terpineno (0,35%), (E)-ß-ocimeno (0,54%), terpinoleno (3,10%), valenceno (0,21%), (Z)-ß-bisaboleno (1,70%) e guaiol (0,29%).

In this work a Piper hispidinervum specie (long pepper), endemic from the Acre state, was grown in Itajaí Valley in Santa Catarina State, and its adaptation was evaluated in relation to chemical composition of essential oil obtained by traditional hydrodistillation and microwave hydrodistillation. The safrole was identified as the major constituent of the essential oil of this specie, which was used as a measure of evaluation of the adaptation capacity of this plant in the southern Brazil, as the proposal includes the evaluation of the usage of this specie as an alternative source of safrole, replacing Sassafras Cinnamon (Ocotea odorifera), widely explored specie in this region until the 90's decade. The plant samples were obtained from different regions of the Acre state and were cultivated at the experimental station of EPAGRI Itajaí - SC. The essential oil of the leaves provided an average content of safrole between 76.6% and 89.9%. The analysis by GC-FID and GC-MS of the oil with higher safrole content, presented the following relative constitution: safrole (89.93%), α-terpinene (0.35%), (E)-ß-ocimene (0.54%), terpinolene (3.10%), valencene (0.21%), (Z)-ß-bisabolene (1.70%) and guaiol (0.29%).

Cryopreservation of Collected Peripheral Blood Hematopoietic Stem Cell Product with 5% DMSO by Adding Nontoxic Natural Cryoprotectants / 대한수혈학회지

BACKGROUND: Cryopreservation of hematopoietic stem cells has become an important process due to the therapeutic protocol, which includes stem cell transplantation after chemotherapy, for many hematological malignancies. The conventional medium contains 10% dimethyl sulfoxide (DMSO) as a cryoprotectant, but this has been reported to be related with many complications. We analyzed the usefulness of trehalose, catalase and zVAD-fmk for cryopreservation along with using a reduced concentration of DMSO to 5%. METHODS: Peripheral blood stem cells were frozen in 10% DMSO as a control and also in 5% DMSO with trehalose and catalase. After 3 weeks of storage in a liquid nitrogen tank, the viability of the thawed hematopoietic stem cells was measured using Trypan blue staining and 7-AAD analysis via conducting flow cytometry. The colony forming potential was assessed using methylcellulose culture. We measured the viability of cells in 5% DMSO medium with or without addition of 30 uM zVAD-fmk right after thawing, and we also did this 6 and 24 hours after incubation. RESULTS: Cryopreserved cells in 5% DMSO with trehalose and catalase showed similar survival (50.42%) compared with the control (49.78%). The viability of cells that were also treated with added zVAD-fmk showed a better result (13.12%) than without it (5.5%) after 24 hours of incubation. Colony forming assay showed similar colony formation in 5% DMSO with the natural cryoprotectants. CONCLUSION: According to the results, lowering the DMSO concentration to 5% is significant and we can expect better cell viability and prevent many side effects of high dose DMSO when adding natural cryprotectants in the cryopreservation medium or by adding caspase-inhibitor right after thawing.

Effects of Lipoxygenase Inhibitor on Diabetic Nephropathy in Rats: Decreasing Proteinuria and Preserving Renal Function / 대한신장학회지

PURPOSE: Oxidative stress leads to an increased production of lipoxygenase derivatives in diabetic nephropathy. Thus, we hypothesized that lipoxygenase inhibitor, nordihydroguaiaretic acid (NDGA), ha the effects of decreasing proteinuria and preserving renal function in streptozotocin (STZ)-induced diabetic rats. METHODS: 45 Sprague-Dawley rats were divided into three groups; (A) treatment with lipoxygenase inhibitor, NDGA in diabetic nephropathy rats, (B) treatment with dimethyl sulfoxide (DMSO) as a vehicle in STZ-induced diabetic rats, (C) normal control group with subcutaneous injection of normal saline. Diabetes was induced by a single intraperitoneal injection of STZ (65 mg/kg) in rats of group A and B. After the 4th week of STZ injection, NDGA (10 mg/kg) and DMSO were given subcutaneously for another 4 weeks in group A and B respectively. RESULTS: The NDGA-treated diabetic rats exhibited significantly decreased urinary albumin excretion. Serum creatinine and blood urea nitrogen concentrations were increased in both group A and B, and tend to be higher in group B than group A. Twenty-four-hour urine creatinine clearances were increased in both group A and B after injection of STZ. Pathologic alterations of kidney were observed after injection of STZ, and then attenuated after administration of NDGA. CONCLUSION: These results suggest the potential of lipoxygenase inhibitor as a complementary therapy for the prevention and treatment of diabetic nephropathy.

Isomerização do óleo essencial de pimenta-longa (Piper hispidinervium C. DC) para a obtenção de isosafrol / Isomerization of essential oil of (Piper hispidinervium) C. DC to obtain isosafrole

As piperáceas brasileiras apresentam propriedades para serem utilizadas, entre outras aplicações, como inseticidas e antifúngicos. Dentre as piperáceas, pode-se citar a Piper hispidinervum C. DC. De suas folhas e dos talos finos extrai-se um óleo essencial rico em safrol, que é utilizado pela indústria como matéria-prima na manufatura, por exemplo, do piperonal. A sequência natural do processo para a obtenção do piperonal é o de submeter o safrol obtido da concentração do óleo essencial à isomerização para seu correspondente isômero estável chamado isosafrol. Apresenta-se neste artigo, por sua vez, a síntese do isosafrol diretamente do óleo essencial de pimenta-longa (Piper hispidinervium C. DC). Este óleo essencial apresenta o safrol como constituinte majoritário, possibilitando a sua isomerização para a produção de isosafrol, que é empregado nas indústrias farmacêuticas e de fragrâncias. O objetivo deste trabalho é o de apresentar a obtenção do isosafrol sem a necessidade da etapa de separação do safrol do óleo essencial de pimenta-longa. Para tanto, foram realizados ensaios de isomerização do óleo essencial, obtendo-se uma solução contendo 79,4 % da mistura cis, trans-isosafrol. Ressalte-se que o óleo essencial continha 86,4 % de safrol, resultando em alto rendimento na sua conversão a cis, trans-isosafrol de 97,1 %.

The Brazilian peppers present proprieties that can be used in fragrance and pharmaceutical industries, for example. One these pepper is long pepper (Piper hispidinervium C. DC.), who its essential oil is rich in safrole. Safrole is an important compound with application in several industries, such as pharmaceutics, cosmetics. After its chemical conversion, for example, it is possible to obtain heliotropin, a fixative agent for fragrances. The sequence to obtain the heliotropin is by safrole isomerization, that, initially, produce isosafrole. In this work, however, presents the synthesis of isosafrole directly from essential oil of Piper hispidinervium C. DC. In this case, the isomerization of essential oil was done getting 79,4 % of cis, trans-isosafrole mixture. The essential oil presented 86,4 % of safrole, resulting in 97,1 % yield to cis, trans-isosafrole.

Atividade inseticida do óleo essencial de pimenta longa (Piper hispidinervum C. DC.) sobre lagarta-do-cartucho do milho Spodoptera frugiperda (J. E. Smith, 1797) (Lepidoptera: Noctuidae) / Insecticidal Activity of Long-pepper essential oil (Piper hispidinervum C. DC.) on fall armyworm Spodoptera frugiperda (J. E. Smith, 1797) (Lepidoptera: Noctuidae)

Os objetivos desta pesquisa foram a obtenção e caracterização do óleo essencial de folhas de pimenta longa Piper hispidinervum, e avaliação de seu efeito no comportamento e/ou mortalidade da lagarta-do-cartucho do milho Spodoptera frugiperda. O óleo essencial foi obtido pela técnica "arraste a vapor d'água", utilizando-se de um aparelho de Clevenger modificado, e posteriormente submetido, à análise por CG-EM e CG. Foram realizados testes de ingestão e contato tópico em lagartas de 1º e 3º ínstar. Os resultados constataram que o óleo essencial de pimenta-longa possui atividade inseticida sobre S. frugiperda, causando redução alimentar e mortalidade, sendo o safrol (82 por cento) seu constituinte majoritário. Verificou-se mortalidade no teste de ingestão em lagartas de 1º ínstar com CL50 = 16,2 mg/mL e para lagartas de 3º ínstar a CL50 = 9,4 mg/mL com redução alimentar CD50 = 0,72 mg/mL; e de toxicidade aguda no teste de contato tópico com DL50 = 277,91 μg/lagarta, após o intervalo de tempo de 96 horas, sendo também observados sintomas de neurotoxicidade, como o efeito knock-down.

The objectives of this research were to evaluate the chemical composition and bioactivity of the essential oil from the long-pepper leaves (Piper hispidinervum) and its effect on the comportment and/or mortality of the fall armyworm (Spodoptera frugiperda). The essential oil was obtained by steam stream distillation, using the modified Clevenger apparatus, the chemical composition was analyzed by GC-MS and GC. Tests of ingestion and topical application in fall armyworm of 1 st and 3 rd ínstar were made. The results showed that the long-pepper essential oil has insecticide activity on S. frugiperda, causing mortality and food reduction, and safrole (82 percent) as its majority constituent. Mortality was found in the ingestion test in larvae of 1 st ínstar LC50 = 16.2 mg/mL for larvae of 3 rd ínstar LC50 = 9.4 mg/mL, with reduced food CD50 = 0.72 mg/mL; and acute toxicity test of the contact topic with LD50 = 277.91 μg/larvae, after an interval of 96 hours, neurotoxicity symptoms were also observed, as knock-down effect.

Enhanced Cardiomyogenic Differentiation of P19 Embryonal Carcinoma Stem Cells

BACKGROUND AND OBJECTIVES: We investigated the effects of different concentrations of serum, 5-azacytidine, and culture time on the cardiomyogenic differentiation of P19 embryonal carcinoma stem cells in the course of developing an efficient protocol for generating the cardiomyogenic lineage. MATERIALS AND METHODS: P19 cells were plated at a density of 1x10(6) cells on 10-cm bacterial dishes for 96 hours in the presence of 1% dimethyl sulfoxide to form embryoid bodies. The embryoid bodies were cultured in medium with 2% or 10% fetal bovine serum for an additional 10 or 15 consecutive days in the presence of 0, 1, or 3 microM 5-azacytidine. RESULTS: Quantitative real-time polymerase chain reaction (PCR) analysis showed that the messenger ribonucleic acid (mRNA) expression of cardiac muscle-specific genes, such as GATA4, alpha-actin, alpha-myosin heavy chain, and cardiac troponin T, were significantly higher in the 15-day culture groups than in the 10-day culture groups. Furthermore, the cardiac muscle-specific genes were expressed more in the high-serum groups compared to the low-serum groups regardless of the culture time. Cardiomyogenic differentiation of the P19 cells was most effective in 1 microM 5-azacytidine regardless of the serum concentrations. In addition, the stimulation effects of 5-azacytidine on cardiomyogenic differentiation were more significant under low-serum culture conditions compared to high-serum culture conditions. Cardiomyogenic differentiation of P19 cells was further confirmed by immunostaining with cardiac muscle-specific antibodies. CONCLUSION:Taken together, these results demonstrated that cardiomyogenic differentiation of P19 cells was enhanced by a combination of different experimental factors.

New neolignan from seed of Myristica fragrans / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents in the nutmeg (seed of Myristica fragrans).</p><p><b>METHOD</b>The chemical constituents were isolated by various column chromatographic methods and structurally elucidated by IR, NMR and MS evidences.</p><p><b>RESULT</b>Fifteen compounds were obtained and identified as myristicin (1), methyleugenol (2), safrole (3), 2, 3-dihydro-7-methoxy-2(3, 4-methylenedioxyphenyl)-3-methyl-5-(E) -propenyl-benzofuran (4), dehydrodiisoeugenol (5), 2, 3-dihydro-7-methoxy-2-(3-methoxy-4, 5-methylenedioxyphenyl) -3-methyl-5-(E)-propenyl-benzofuran (6), erythro-2-(4-allyl-2, 6-dimethoxyphenoxy)-1-(3, 4-dimetho- xyphenyl) propane (7), erythro-2-(4-allyl-2, 6-dimethoxyphenoxy)-1-(3, 4, 5-trimethoxyphenyl) propane (8), erythro-2-(4-allyl-2, 6-dimethoxyphenoxy)-1-(3, 4-dimethoxyphenyl) propan-1-ol acetate (9), erythro-2-(4-allyl-2, 6-dimethoxyphenoxy)-1-(3, 4-dimethoxyphenyl) propan-1-ol (10), erythro-2-(4-allyl-2, 6-dimethoxyphenoxy)-1-(3, 4, 5-trimethoxyphenyl) propan-1-ol (11), 5-methoxy-dehydrodiisoeugenol (12), erythro-2-(4-allyl-2, 6-dimethoxyphenoxy)-1-(4-hydroxy-3-methoxyphenyl)-propan-1-ol (13), guaiacin (14) and threo-2-(4-allyl-2, 6-dimethoxyphenoxy)-1-(3-methoxy-5-hydroxy-phenyl) propan-1-ol (15).</p><p><b>CONCLUSION</b>Compound 15 is a new compound and named myrisisolignan. Compound 7 is isolated from the genus Myristica for the first time.</p>

Efeito do óleo essencial de pimenta longa (Piper hispidinervum C. DC) e do emulsificante Tween® 80 sobre o crescimento micelial de Alternaria alternata (Fungi: Hyphomycetes) / The effect of Long-pepper essential oil (Piper hispidinervum C. DC.) and of Tween®; 80 emulsifier on the mycelial growth of Alternaria alternate (Fungi: Hyphomycetes)

Este trabalho objetivou avaliar o efeito do óleo essencial de folhas de pimenta longa (Piper hispidinervum C. DC), sobre o crescimento micelial de Alternaria alternata e a análise da influência da concentração do emulsificante Tween® 80 no controle deste fitopatógeno. O óleo essencial foi obtido pela técnica "arraste a vapor d'água", utilizando-se aparelho de Clevenger modificado, e posteriormente submetido, à análise por cromatografia em fase gasosa acoplada a espectrômetro de massas CG-EM a CG. Para os ensaios biológicos, o método foi o bioanalítico in vitro observando-se o crescimento ou inibição do micélio de A. alternata no meio de cultura BDA na presença de diferentes concentrações do óleo essencial (0, 100, 250, 500 e 1000 mgL-1 ) sob diferentes concentrações de Tween® 80. Adotou-se esquema fatorial com quatro repetições, em delineamento inteiramente casualizado (DIC). Observou-se que o óleo essencial de pimenta longa apresentou inibição sobre o crescimento micelial do fungo A. alternataem todas as concentrações analisadas, sendo que na concentração de 1000 mgL-1 esta inibição foi de 100 por cento , e a porcentagem de emulsificante (Tween® 80), influenciou na atividade fungitóxica das concentrações de 250 mgL -1 e 500 mgL -1 do óleo essencial.

The object of this work was to evaluate the effectof essential oils from Long-pepper leaves Piper hispidinervum on the mycelial growth of Alternaria alternata and to analyze the influence of emulsifier Tween® 80 concentration in the control of this phytopathogen. The essential oil was obtained by steam stream distillation, using a modified Clevenger apparatus, and the chemical composition was analyzed by gas chromatography coupled to mass spectroscopy GC-MS and GC. For biological tests, the bioanalytical in vitro method was used, observing the growth or inhibition of such phytopathogen in BDA culture media, in the presence of different essential oil concentrations (0, 100, 250, 500, e 1000 mgL -1) under different Tween® 80 concentrations. Factorial scheme with four repetitions, in entirely randomized outline was adopted. The essential oil of Long-pepper inhibited the micellial growth of the A. alternata fungus at all of the concentrations tested. A 100 percent inhibition at the 1000 mgL -1 concentration was observed, and the percentage of emulsifier (Tween® 80) influenced the fungitoxic activity at the 250 mgL-1 and 500 mgL -1 concentrations of the essential oil.

GC-MS analysis of essential oil from nutmeg processed by different traditional methods / 中国中药杂志

<p><b>OBJECTIVE</b>To analyze the chemical components of the essential oil extracted from the seeds of Myristica fragrans (nutmeg) processed by different methods (steamed with water steam, roasted with flour, sauted with flour, roasted with talcum powder, roasted with loess, and roasted with bran) and to provide quality control foundations in the sciences.</p><p><b>METHOD</b>The essential oil was extracted by steam distillation and separated with GC capillary column. The relative content of every compound was determined with area normalization method and the structures were elucidated by GC-MS technique.</p><p><b>RESULT</b>Fifty-eight to one hundred and four of chromatographic peaks were detected, among them seventy-six compounds accounting for 98.32% to 99.99% of the total essential oil in nutmeg were identified, which were composed of 69.15% to 97.24% for monoterpenoids and 2.06% to 25.51% for aromatic compounds of the total essential oil, respectively.</p><p><b>CONCLUSION</b>It was shown that monoterpenoids and their derivatives were main composition, and aromatic compounds were secondary composition in the total essential oil of nutmeg grows in Indonesia and processed by different traditional methods on the basis of theory of traditional Chinese medicine. In addition, it was suggested that we should be careful to use processed nutmeg owing to contain safrole and a-asarone induced genetoxicity in animals and mutagenicity in the Ames Salmonella assay, and myristicin and elemicin induced narcotism in human. The processed method roasted with bran for nutmeg may be better and will be developed.</p>

Comparing analysis of components in volatile oils of nutmeg and prepared nutmeg by GC-MS / 中国中药杂志

<p><b>OBJECTIVE</b>To compare components in volatile oils of nutmeg and prepared nutmeg.</p><p><b>METHOD</b>Volatile oil from nutmeg and prepared nutmeg were extracted by vapor distillation. The chemical components in two kinds of volatile oils were determined and indentified by GC-MS.</p><p><b>RESULT</b>The change in quantity and quality of components in volatile oils were observed after processing. 13 new components occurred and 4 components disappeared in volatile oils after processing. The contents of methyleugenol and methylisoeugenol that are active ingredients were increased. The contents of myristicin and safrol that are toxic ingredients in volatile oils were decreased.</p><p><b>CONCLUSION</b>The processing method of nutmeg by soaking with water and roasting with bran is scientific.</p>

Study on GC fingerprint of the constituents in Herba Asari / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a method for GC fingerprint determination of the chemical constituents in Herba Asari.</p><p><b>METHOD</b>GC and GC-MS were used to optimize the fingerprint determination method, and identify the main peaks in the GC fingerprint.</p><p><b>RESULT</b>A preferable method for GC fingerprint determination of the chemical constituents in Herba Asari was established.</p><p><b>CONCLUSION</b>A general acquaintance of the chemical constituents in Herba Asari can be obtained by using the preferable GC fingerprint determination method, which is useful for quality evaluation of the crude drug of Herba Asari.</p>

Estratégias em química medicinal para o planejamento de fármacos / Strategies in medicinal chemistry to the discovery of new lead-compounds

Este artigo ilustra a aplicação das estratégias de hibridação molecular e bioisosterismo no planejamento estrutural de novos candidatos a protótipos de fármacos com propriedades analgésicas, antiiinflamatórias e antitrombóticas, com base no mecanismo de ação pretendido, explorando, inclusive, produto natural brasileiro abundante como padrão molecular básico...

Quaternary ammonium salt derivatives of allylphenols with peripheral analgesic effect

Ammonium salt derivatives of natural allylphenols were synthesized with the purpose of obtaining potential peripheral analgesics. These drugs, by virtue of their physicochemical properties, would not be able to cross the blood brain barrier. Their inability to enter into the central nervous system (CNS) should prevent several adverse effects observed with classical opiate analgesics (Ferreira et al., 1984). Eugenol (1) O-methyleugenol (5) and safrole (9) were submitted to nitration, reduction and permethylation, leading to the ammonium salts 4, 8 and 12. Another strategy applied to eugenol (1), consisting in its conversion to a glycidic ether (13), opening the epoxide ring with secondary amines and methylation, led to the ammonium salts 16 and 17. All these ammonium salts showed significant peripheral analgesic action, in modified version of the Randall-Sellito test (Ferreira et al. 1978), at non-lethal doses. The ammonium salt 8 showed an activity comparable to that of methylnalorphinium, the prototype of an ideal peripheral analgesic (Ferreira et al., 1984).

Estudo de espectros de ressonância magnética nuclear protônica e de carbono 13 de uma série de derivados de alilfenóis / Study of proton and carbon 13 nuclear magnetic ressonance spectra of an allylphenol derivatives series

Sais de amônio quaternários derivados de alilfenóis naturais foram sintetizados com o objetivo de se obter analgésicos periféricos, Eugenol, Ometileugenol e safrol foram submetidos a nitraçäo, reduçäo e permetilaçäo. As aminas também foram transformadas nos respectivos cloridratos. Os espectros de RMN1 H dos nitrocompostos, aminas, cloridratos de aminas, sais de amônio quaternários e dimetilaminas foram analisados, assim como os espectros de RMN13 C dos nitrocompostos e dos sais de amônio quaternários.

Anti-inflammatory preoperties of new bioisosteres of indomethacim synthesized from safrole which are sulindac analogues

The anti-inflammatory activities of new compounds (I, II, II and IV) synthesized in 30% overall yield from the abundant natural product safrole, the principal chemical constituent of the oil of sassafras (Ocotea pretiosa, Lauraceae), were determined in mice. The synthesis of these new indenyl-acetic acids (I and II) and indenyl-propionic acids (III and IV) was based on the minimal structural features of non-steroid anti-inflammatory agents of the aryl- or heteroarylcarboxylic acid group. The compounds exhibited potencies 4- to 10-fold less than that of indometacin in inhibiting carrageenan-induced hindpaw edema. In contrast, like sulindac, all the new compounds were more potent than indomethacin in antagonizing writhing pain and increased vascular permeability caused by acetic acid. The results confirm the anticipated bioisosteric relationship between these synthetic derivatives, designed as sulindac analogues, and the classical non-steroidal anti-inflammatory agent, indomethacin