Sesquiterpene Lactones with Anti-Hepatitis C Virus Activity Using Molecular Descriptors

Hepatitis C is a worldwide public health problem. The available therapies are limited by their partial effectiveness and with meaningful side-effects. Sesquiterpene lactones (SLs) are a group of natural products with a wide variety of chemical structures and biological activities associated. There are few studies about the influence of the molecular structure of SLs for the anti-hepatitis C virus activity. In the present work, SLs are investigated in a subgenomic RNA replicon assay system and were analyzed using multiple linear regression along with self-organizing maps with DRAGON descriptors in order to identify the structural requirements for their biological activity and to predict the inhibitory potency of SLs. Characteristics such as stereochemistry and electronic effects demonstrated to be important for their anti-HCV activity, and the SOM produced a clear separation betwenn active and inactive compounds. Therefore, it is possible to use this map as a filter for virtual screening to predict the anti-HCV activity of SLs.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq

Structure–Activity Relationship of Piplartine and Synthetic Analogues against Schistosoma mansoni and Cytotoxicity to Mammalian Cells

Schistosomiasis, caused by helminth flatworms of the genus Schistosoma, is an infectious disease mainly associated with poverty that affects millions of people worldwide. Since treatment for this disease relies only on the use of praziquantel, there is an urgent need to identify new antischistosomal drugs. Piplartine is an amide alkaloid found in several Piper species (Piperaceae) that exhibits antischistosomal properties. The aim of this study was to evaluate the structure–function relationship between piplartine and its five synthetic analogues (19A, 1G, 1M, 14B and 6B) against Schistosoma mansoni adult worms, as well as its cytotoxicity to mammalian cells using murine fibroblast (NIH-3T3) and BALB/cN macrophage (J774A.1) cell lines. In addition, density functional theory calculations and in silico analysis were used to predict physicochemical and toxicity parameters. Bioassays revealed that piplartine is active against S. mansoni at low concentrations (5⁻10 µM), but its analogues did not. In contrast, based on 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry assays, piplartine exhibited toxicity in mammalian cells at 785 µM, while its analogues 19A and 6B did not reduce cell viability at the same concentrations. This study demonstrated that piplartine analogues showed less activity against S. mansoni but presented lower toxicity than piplartine.This work was partially supported by grants from the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP grant number 2016/22488-3), the authors are grateful to CAPES/FAPEPI for the scholarship granted. ACM is grateful to FAPESP (2014/02282-6). YPM is grateful to CNPq (Grant 302674/2010-1). Financial Support: FCT (Fundação para Ciência e Tecnologia) by grant no. UID/MULTI/04378/2013 POCI/01/0145/FEDER/007728, Fapesp (2014/50316-7). The work at UCIBIO/Requimte was supported by the Fundação para a Ciência e a Tecnologia (FCT) with financial support from FCT/MEC through national funds and co-financed by FEDER, under the Partnership Agreement PT2020. Alexandra Plácido is grateful to FCT by her grant SFRH/BD/97995/2013, financed by POPH–QREN–Tipologia 4.1–Formação Avançada, subsidized by Fundo Social Europeu and Ministério da Ciência, Tecnologia e Ensino Superior.info:eu-repo/semantics/publishedVersio

Use of Multivariate Statistics in the Chemosystematics of Asteraceae family and its Heliantheae tribe

Este trabalho análise as ocorrências de 12 classes de substâncias (monoterpenos, sesquiterpenos, lactonas sesquiterpênicas, diterpenos, triterpenos, cumarinas, flavonóides, poliacetilenos, benzofuranos, benzopiranos, acetofenonas e fenilpropanóides) na família Asteraceae e na sua tribo Heliantheae. Pretende-se demonstrar a existência de correlações na produção de metabólitos secundários em níveis taxonômicos baixos (tribos, subtribos e gêneros). Utilizou-se um banco de dados com cerca de 36.000 ocorrências das principais substâncias isoladas em plantas da família. O estudo do equilíbrio químico na produção de metabólitos secundários foi feito utilizando-se Regressão Linear Múltipla. As afinidades entre os grupos com base na sua Química foram pesquisadas por vários métodos tais como: Análise de componentes principais, Análise de Cluster e Análises cladísticas. Observou-se também o grau de oxidação médio de vários metabólitos e sua utilidade como ferramenta em análises quimiotaxonômicas. Foi possível mostrar a existência de um equilíbrio na produção das 12 classes de metabólitos em níveis das tribos e subtribos. Mas, no nível dos gêneros um equilíbrio moderado foi encontrado. Também foi possível mostrar a existência de um equilíbrio oxidativo em vários níveis (tribos, subtribos). No nível dos gêneros nenhum equilíbrio foi encontrado utilizando-se o parâmetro passo oxidativo. Foi possível agrupar algumas das subfamílias de Asteraceae segundo Bremer e subtribos da tribo Heliantheae segundo Stuessy usando Análises de componentes principais e Análise de ClusterThis work analyse the occurrence of 12 classes of substances (monoterpenes, sesquiterpenes, sesquiterpene lactones, diterpenes, triterpenes, coumarins, flavonoids, polyacetylenes, Benzofurans, benzopyrans, acetophenones and phenylpropanoids) in the Asteraceae family and its Heliantheae tribe. This study intends to demonstrate the existence of correlations in the production of secondary metabolites in lower taxonomic levels (tribes, subtribes and genera). We used a database of about 36,000 occurrences of the main substances isolated from the plant family. The study of chemical equilibrium in the production of secondary metabolites was done using Multiple Linear Regression. The affinities between the groups based on their chemistry were investigated by various methods such as principal component analysis, Cluster and cladistic analysis. There was also the average degree of oxidation of various metabolites and their usefulness as a tool in chemotaxonomic analysis. It was possible to show the existence of a balance in the production of 12 classes of metabolites in the levels of the tribes and subtribes. But the level of the genus balance was found moderate. It was also possible to show the existence of an oxidative equilibrium in various levels (tribes, subtribes). The level of genus balance was not found using the parameter oxidation step. We could group some of the subfamilies of Asteraceae according to Bremer and the subtribes of Heliantheae according to Stuessy using the principal component analysis and Cluster Analysi

Use of Multivariate Statistics in the Chemosystematics of Asteraceae family and its Heliantheae tribe

Este trabalho análise as ocorrências de 12 classes de substâncias (monoterpenos, sesquiterpenos, lactonas sesquiterpênicas, diterpenos, triterpenos, cumarinas, flavonóides, poliacetilenos, benzofuranos, benzopiranos, acetofenonas e fenilpropanóides) na família Asteraceae e na sua tribo Heliantheae. Pretende-se demonstrar a existência de correlações na produção de metabólitos secundários em níveis taxonômicos baixos (tribos, subtribos e gêneros). Utilizou-se um banco de dados com cerca de 36.000 ocorrências das principais substâncias isoladas em plantas da família. O estudo do equilíbrio químico na produção de metabólitos secundários foi feito utilizando-se Regressão Linear Múltipla. As afinidades entre os grupos com base na sua Química foram pesquisadas por vários métodos tais como: Análise de componentes principais, Análise de Cluster e Análises cladísticas. Observou-se também o grau de oxidação médio de vários metabólitos e sua utilidade como ferramenta em análises quimiotaxonômicas. Foi possível mostrar a existência de um equilíbrio na produção das 12 classes de metabólitos em níveis das tribos e subtribos. Mas, no nível dos gêneros um equilíbrio moderado foi encontrado. Também foi possível mostrar a existência de um equilíbrio oxidativo em vários níveis (tribos, subtribos). No nível dos gêneros nenhum equilíbrio foi encontrado utilizando-se o parâmetro passo oxidativo. Foi possível agrupar algumas das subfamílias de Asteraceae segundo Bremer e subtribos da tribo Heliantheae segundo Stuessy usando Análises de componentes principais e Análise de ClusterThis work analyse the occurrence of 12 classes of substances (monoterpenes, sesquiterpenes, sesquiterpene lactones, diterpenes, triterpenes, coumarins, flavonoids, polyacetylenes, Benzofurans, benzopyrans, acetophenones and phenylpropanoids) in the Asteraceae family and its Heliantheae tribe. This study intends to demonstrate the existence of correlations in the production of secondary metabolites in lower taxonomic levels (tribes, subtribes and genera). We used a database of about 36,000 occurrences of the main substances isolated from the plant family. The study of chemical equilibrium in the production of secondary metabolites was done using Multiple Linear Regression. The affinities between the groups based on their chemistry were investigated by various methods such as principal component analysis, Cluster and cladistic analysis. There was also the average degree of oxidation of various metabolites and their usefulness as a tool in chemotaxonomic analysis. It was possible to show the existence of a balance in the production of 12 classes of metabolites in the levels of the tribes and subtribes. But the level of the genus balance was found moderate. It was also possible to show the existence of an oxidative equilibrium in various levels (tribes, subtribes). The level of genus balance was not found using the parameter oxidation step. We could group some of the subfamilies of Asteraceae according to Bremer and the subtribes of Heliantheae according to Stuessy using the principal component analysis and Cluster Analysi

Synthesis, biological activities and structure-activity relationship study of piperamides

As estruturas e propriedades biológicas das amidas piplartina e a piperina, isoladas respectivamente de Piper tuberculatum e P. nigrum, inspiraram a síntese de 89 derivados e 7 esters estruturalmente relacionadas. As preparações envolveram metodologias tradicionais e os compostos purificados tiveram suas estruturas caracterizadas por análises espectroscópicas e espectrométricas. Os estudos de fragmentação por IE e IES indicaram a clivagem preferencial da ligação N-CO no caso das cinamamidas, dienamidas e cinamimidas. Estudos computacionais envolvendo afinidade protônica e energias de ligação confirmaram a fragmentação preferencial da ligação amídica para as amidas. A citotoxicidade de 89 substâncias foi avaliada contra três células leucêmicas (K562, Nalm6 e Raji) e a partir dos valores de IC50 foram realizados estudos de relação estrutura-atividade (SAR). As linhagens K562 e a Nalm6 foram a mais resistente e vulnerável, respectivamente, e as amidas piplartina (1a), N-Ciclohexil-N-(ciclohexilcarbamoil)-3-(3,4,5-trimetoxifenil)propanamida (1n), e (E)-N,N-dibutil-3-(3,4-dimetoxifenil)acrilamida (13h) foram as mais ativas com IC50 de 0,34 µM; 0,84 µM e 1,88 µM contra K562 e (E)-N-ciclohexil-N-(ciclohexilcarbamoil)-3-(3,4-dimetoxifenil)acrylamida (13i) com IC50 de 0,98 µM contra Nalm6. A avaliação de atividade leishmanicida de 18 substâncias não se mostrou promissora. As abordagens qualitativas e quantitativas foram feitas baseadas nos descritores moleculares gerados pelo programa VolSurf+. A partir de métodos quimiométricos tais com PLS, algoritmo genético, árvores de decisão foi possível gerar modelos para correlacionar às propriedades moleculares com a atividade biológica. As propriedades de absorção, distribuição, metabolismo e excreção e os equilíbrios entres as regiões hidrofílicas e hidrofóbicas foram importantes para atividade citotóxica. O estudo de ancoragem molecular mostrou que as amidas (E)-N,N-dibutil-3-(3,4,5-trimetoxifenil)acrilamida (1l), 1n, (E)-3-(4-clorofenil)-N-ciclohexil-N-(ciclohexilcarbamoil)acrilamida (5a), 13h e 13i podem atuar como inibidores das histonas desacetilases particularmente HDAC4 e HDAC8.The structures and biological properties of the amides piplartine and piperine isolated from Piper tuberculatum and P. nigrum respectively, inspired the synthesis of derivatives 89 and 7 esters structurally related. Their preparations were achieved using classical procedures and the purified amides were submitted to spectroscopic and spectrometric characterization. The study of fragmentation process by EI and ESI suggested the preferential cleavage of the N-CO bond of cinnamamides, dienamides and cinnamimides. The cytotoxicity of 89 compounds was evaluated against three leukemic cells (K562, Nalm6 and Raji) and based on IC50 values the structure-activity relationship (SAR) was performed. While the K562 and Nalm6 cells were the more resistant and more sensitive, respectively, the amides piplartine (1a), N-cyclohexyl-N-(ciclohexylcarbamoyl)-3-(3,4,5-trimethoxyphenyl)propanamide (1n) and (E)-N,N-dibutyl-3-(3,4-dimethoxyphenyl)acrylamide (13h) were in general the most active with IC50 of 0.34 µM, 0.84 µM and 1.88 µM against K562 and (E)-N-cyclohexyl-N-(ciclohexylcarbamoyl)-3-(3,4-dimethoxyphenyl)acrylamide (13i) with IC50 of 0.98 µM against Nalm6. The evaluation of leishmanicidal activity of 18 substances was also performed but was not promising. Qualitative and quantitative approaches were made based on molecular descriptors generated by VolSurf+ program. The chemometric methods such as PLS, genetic algorithm, decision trees generated models to correlate molecular properties with the biological activity. The absorption, distribution, metabolism and excretion properties and a balance between hydrophilic and hydrophobic moieties of the amides were important for an optimized activity. The molecular docking revealed that amides such as (E)-N,N-dibutyl-3-(3,4,5-trimethoxyphenyl)acrylamide (1l), 1n, (E)-3-(4-chlorophenyl)-N-cyclohexil-N-(cyclohexylcarbamoyl)acrylamide (5a), 13h and 13i have potential to act as possible inhibitors of histone deacetylase proteins particularly HDAC4 and HDAC8

Effect of piplartine and cinnamides on Leishmania amazonensis, Plasmodium falciparum and on peritoneal cells of Swiss mice

Context: Plants of the Piperaceae family produce piplartine that was used to synthesize the cinnamides. Objective: To assess the effects of piplartine (1) and cinnamides (2–5) against the protozoa responsible for malaria and leishmaniasis, and peritoneal cells of Swiss mice. Materials and methods: Cultures of Leishmania amazonensis, Plasmodium falciparum-infected erythrocytes, and peritoneal cells were incubated, in triplicate, with different concentrations of the compounds (0 to 256 μg/mL). The inhibitory concentration (IC50) in L. amazonensis and cytotoxic concentration (CC50) in peritoneal cell were assessed by the MTT method after 6 h of incubation, while the IC50 for P. falciparum-infected erythrocytes was determined by optical microscopy after 48 or 72 h of incubation; the Selectivity Index (SI) was calculated by CC50/IC50. Results: All compounds inhibited the growth of microorganisms, being more effective against P. falciparum after 72 h of incubation, especially for the compounds 1 (IC50 = 3.2 μg/mL) and 5 (IC50 = 6.6 μg/mL), than to L. amazonensis (compound 1 = 179.0 μg/mL; compound 5 = 106.0 μg/mL). Despite all compounds reducing the viability of peritoneal cells, the SI were 10 for the piplartine (>37.4) and cinnamides 4 (>10.7) and 5 (= 38.4). Discussion and conclusion: The potential of piplartine and cinnamides 4 and 5 in the treatment of malaria suggest further pre-clinical studies to evaluate their effects in murine malaria and to determine their mechanisms in cells of the immune system

Morphological changes in <i>D. rerio</i> during the 48 hours of exposure to piplartine.

<p><b>A</b>) Leakage of the ocular pigment caused by 1.8 ppm piplartine, <b>B</b>) tissue alterations on the head and mouth caused by 1.6 µg/ml piplartine, <b>C</b>) exophthalmia and hemorrhage caused by 1.4 µg/ml piplartine and D) control group.</p

Mortality of <i>B. glabrata</i> adults exposed to methanolic extracts of different organs of <i>Piper tuberculatum</i>.

<p>n = 30 adult snails.</p><p>Values were obtained at the end of the 7^th day of observation.</p