Biodereplication of antiplasmodial extracts: application of the amazonian medicinal plant piper coruscans kunth

Improved methodological tools to hasten antimalarial drug discovery remain of interest, especially when considering natural products as a source of drug candidates. We propose a biodereplication method combining the classical dereplication approach with the early detection of potential antiplasmodial compounds in crude extracts. Heme binding is used as a surrogate of the antiplasmodial activity and is monitored by mass spectrometry in a biomimetic assay. Molecular networking and automated annotation of targeted mass through data mining were followed by mass-guided compound isolation by taking advantage of the versatility and finely tunable selectivity offered by centrifugal partition chromatography. This biodereplication workflow was applied to an ethanolic extract of the Amazonian medicinal plant Piper coruscans Kunth (Piperaceae) showing an IC50 of 1.36 ug/mL on the 3D7 Plasmodium falciparum strain. It resulted in the isolation of twelve compounds designated as potential antiplasmodial compounds by the biodereplication workflow. Two chalcones, aurentiacin (1) and cardamonin (3), with IC50 values of 2.25 and 5.5 uM, respectively, can be considered to bear the antiplasmodial activity of the extract, with the latter not relying on a heme-binding mechanism. This biodereplication method constitutes a rapid, efficient, and robust technique to identify potential antimalarial compounds in complex extracts such as plant extracts

Cannabinoids vs. whole metabolome: Relevance of cannabinomics in analyzing Cannabis varieties

International audienceCannabis sativa has a long history of domestication both for its bioactive compounds and its fibers. This has produced hundreds of varieties, usually characterized in the literature by chemotypes, with Δ9-THC and CBD content as the main markers. However, chemotyping could also be done based on minor compounds (phytocannabinoids and others). In this work, a workflow, which we propose to name cannabinomics, combines mass spectrometry of the whole metabolome and statistical analysis to help differentiate C. sativa varieties and deciphering their characteristic markers. By applying this cannabinomics approach to the data obtained from 20 varieties of C. sativa (classically classified as chemotype I, II, or III), we compared the results with those obtained by a targeted quantification of 11 phytocannabinoids. Cannabinomics can be considered as a complementary tool for phenotyping and genotyping, allowing the identification of minor compounds playing a key role as markers of differentiation

Chemical characterization and oxidative stability of the oils from three morphotypes of <i>Mauritia flexuosa</i> L.f, from the Peruvian Amazon

Three morphotypes of aguaje <i>Mauritia flexuosa</i> were tested, classified by the color of their mesocarpium: “color”, “shambo” and “amarillo”, collected from different areas near the city of Iquitos, Peru. Also, physical-chemical analyses of the mesocarpium were performed, such as the characterization of fatty acids by gas chromatography, determination of β- carotene y α – tocopherol by high efficiency liquid chromatography system in normal and reverse phase and the determination of oxidation induction time in the Rancimat apparatus. Proximate, mineral and fatty acid analyses were done on the seeds. The aguaje mesocarpium is rich in oleic oil (“amarillo”: 75.63% ± 0.31), β-carotene (“amarillo”: 342.42ug/g ± 0.71) and α- tocopherol (“color”: 685.81mg/L ± 1.04), plus the morphotype “color” has a superior oxidation induction time compared to other morphotypes with 6.91 ± 0.01. The aguaje seed contains significant amounts of ω6 (linoleic oil) in “shambo” with 36.04 ± 0.09%. The results indicate that these oils, regardless their classification, contain important chemical compounds that give them a special nutritive value.<br><br>Se estudiaron tres morfotipos de aguaje <i>Mauritia flexuosa</i> L. f., clasificados por el color del mesocarpio: “color”, “shambo” y “amarillo”, recogidos de localidades cercanas a la ciudad de Iquitos- Perú. Del mismo modo a partir del mesocarpio se realizaron los análisis fisicoquímicos, la caracterización de los ácidos grasos por cromatografía gaseosa, la determinación de β-caroteno y α– tocoferol por cromatografía líquida de alta eficiencia (HPLC) fase reversa y normal y la determinación del tiempo de inducción de la oxidación en el equipo de Rancimat. A partir de la semilla se realizaron los análisis centesimales, de minerales y ácidos grasos. Los mesocarpios de aguaje son ricos en ácido oleico (amarillo: 75.63% ± 0.31), β-caroteno (amarillo: 342.42 ug/g ± 0.71) y α- tocoferol (color: 685.81mg/L ± 1.04), y además el morfotipo “color” tiene un tiempo de inducción superior a los otros morfotipos con 6.91 ± 0.01, asimismo en la semilla de aguaje se encuentran cantidades considerables de ω6 (ácido linoleico) en “shambo” con 36.04 ± 0.09%. Los resultados obtenidos nos indican que estos aceites independientemente de la clasificación que se le asigne contienen importantes compuestos químicos que le brindan un altor valor nutricional

CARACTERIZACIÓN QUÍMICA DE NUEVE ACCESIONES DE Plukenetia volubilis L. DE LOS DEPARTAMENTOS DE LORETO Y SAN MARTÍN

Se realizó la caracterización química de las semillas de nueve accesiones de sacha inchi Plukenetia volubilis L. del “Centro de Investigaciones de Pucayacu” (Banco de germoplasma - Instituto de Investigaciones de la Amazonía Peruana) en Tarapoto, San Martín. Esta caracterización fue basada en análisis centesimales, análisis fisicoquímicos de aceites, determinación de minerales, caracterización de ácidos grasos e identificación de aminoácidos esenciales. Se determinó que las mejores accesiones en rendimiento de aceite fueron las accesiones 4 y 2 con 20.12% y 20.23% respectivamente. En cuanto a minerales las mejores accesiones fueron (en miligramos por 100g de muestra seca) el 4 (K=892.93; Na=238.02; Zn=16.43) y el 5 (K=863.65; Zn=15.95; Mg=340.15). En tanto que en ácidos grasos los mayores valores de ù-3, fueron encontrados en la accesión 18 (47.91%) y ù-6 en la accesión 6 (41.09%). Todos los resultados encontrados fueron significativos estadísticamente (test Tukey), permitiéndonos determinar que existen diferencias concretas en la características químicas entre las accesiones estudiadas

Metabotyping of Andean pseudocereals and characterization of emerging mycotoxins

International audiencePseudocereals are best known for three crops derived from the Andes: quinoa (Chenopodium quinoa), canihua (C. pallidicaule), and kiwicha (Amaranthus caudatus). Their grains are recognized for their nutritional benefits; however, there is a higher level of polyphenism. Meanwhile, the chemical food safety of pseudocereals remains poorly documented. Here, we applied untargeted and targeted metabolomics approaches by LC-MS to achieve both: i) a comprehensive chemical mapping of pseudocereal samples collected in the Andes; and ii) a quantification of their contents in emerging mycotoxins. An inventory of the fungal community was also realized to better know the fungi present in these grains. Metabotyping permitted to add new insights into the chemotaxonomy of pseudocereals, confirming the previously established phylotranscriptomic clades. Sixteen samples from Peru (out of 27) and one from France (out of one) were contaminated with Beauvericin, an emerging mycotoxin. Several mycotoxigenic fungi were detected, including Aspergillus sp., Penicillium sp., and Alternaria sp

Chemical modulation of the metabolism of an endophytic fungal strain of Cophinforma mamane using epigenetic modifiers and amino-acids

Endophytic fungi are capable of producing a great diversity of bioactive metabolites. However, the presence of silent and lowly expressed genes represents a main challenge for the discovery of novel secondary metabolites with different potential uses. Epigenetic modifiers have shown to perturb the production of fungal metabolites through the induction of silent biosynthetic pathways leading to an enhanced chemical diversity. Moreover, the addition of bioprecursors to the culture medium has been described as a useful strategy to induce specific biosynthetic pathways. The aim of this study was to assess the effects of different chemical modulators on the metabolic profiles of an endophytic fungal strain of Cophinforma mamane (Botryosphaeriaceae), known to produce 3 thiodiketopiperazine (TDKP) alkaloids (botryosulfuranols A-C), previously isolated and characterized by our team. Four epigenetic modifiers, 5-azacytidine (AZA), sodium butyrate (SB), nicotinamide (NIC), homoserine lactone (HSL) as well as 2 amino acids, L-phenylalanine and L-tryptophan, as bioprecursors of TDKPs, were used. The metabolic profiles were analysed by UHPLC-HRMS/MS under an untargeted metabolomics approach. Our results show that the addition of the two amino acids in C. mamane culture and the treatment with AZA significantly reduced the production of the TDKPs botryosulfuranols A, B and C. Interestingly, the treatment with HSL significantly induced the production of different classes of diketopiperazines (DKPs). The treatment with AZA resulted as the most effective epigenetic modifier for the alteration of the secondary metabolite profile of C. mamane by promoting the expression of cryptic genes

Anti-protozoal and anti-fungal evaluation of 3,5-disubstituted 1,2-dioxolanes

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Metabolomic approach of the antiprotozoal activity of medicinal Piper species used in Peruvian Amazon

International audienceEthnopharmacological relevance : In the Peruvian Amazon as in the tropical countries of South America, the use of medicinal Piper species (cordoncillos) is common practice, particularly against symptoms of infection by protozoal parasites. However, there is few documented information about the practical aspects of their use and few scientific validation. The starting point of this work was a set of interviews of people living in six rural communities from the Peruvian Amazon (Alto Amazonas Province) about their uses of plants from Piper genus: one community of Amerindian native people (Shawi community) and five communities of mestizos. Infections caused by parasitic protozoa take a huge toll on public health in the Amazonian communities, who partly fight it using traditional remedies. Validation of these traditional practices contributes to public health care efficiency and may help to identify new antiprotozoal compounds.Aims of study : To record and validate the use of medicinal Piper species by rural people of Alto Amazonas Province (Peru) and annotate active compounds using a correlation study and a data mining approach.Materials and methods : Rural communities were interviewed about traditional medication against parasite infections with medicinal Piper species. Ethnopharmacological surveys were undertaken in five mestizo villages, namely: Nueva Arica, Shucushuyacu, Parinari, Lagunas and Esperanza, and one Shawi community (Balsapuerto village). All communities belong to the Alto Amazonas Province (Loreto region, Peru). Seventeen Piper species were collected according to their traditional use for the treatment of parasitic diseases, 35 extracts (leaves or leaves and stems) were tested in vitro on P. falciparum (3D7 chloroquine-sensitive strain and W2 chloroquine-resistant strain), Leishmania donovani LV9 strain and Trypanosoma brucei gambiense. Assessments were performed on HUVEC cells and RAW 264.7 macrophages. The annotation of active compounds was realized by metabolomic analysis and molecular networking approach.Results : Nine extracts were active (IC50 ≤ 10 μg/mL) on 3D7 P. falciparum and only one on W2 P. falciparum, six on L. donovani (axenic and intramacrophagic amastigotes) and seven on Trypanosoma brucei gambiense. Only one extract was active on all three parasites (P. lineatum). After metabolomic analyses and annotation of compounds active on Leishmania, P. strigosum and P. pseudoarboreum were considered as potential sources of leishmanicidal compounds.Conclusions : This ethnopharmacological study and the associated in vitro bioassays corroborated the relevance of use of Piper species in the Amazonian traditional medicine, especially in Peru. A series of Piper species with few previously available phytochemical data have good antiprotozoal activity and could be a starting point for subsequent promising work. Metabolomic approach appears to be a smart, quick but still limited methodology to identify compounds with high probability of biological activity

Pharmacological validation of Solanum mammosum L. as an anti-infective agent: Role of solamargine

International audienceEthnopharmacological relevanceFungal and bacterial infections remain a major problem worldwide, requiring the development of effective therapeutic strategies. Solanum mammosum L. (Solanaceae) (“teta de vaca”) is used in traditional medicine in Peru to treat fungal infections and respiratory disorders via topical application. However, the mechanism of action remains unknown, particularly in light of its chemical composition.Materials and methodsThe antifungal activity of TDV was determined against Trichophyton mentagrophytes and Candida albicans using bioautography-TLC-HRMS to rapidly identify the active compounds. Then, the minimum inhibitory concentration (MIC) of the fruit crude extract and the active compound was determined to precisely evaluate the antifungal activity. Additionally, the effects of the most active compound on the formation of Pseudomonas aeruginosa biofilms and pyocyanin production were evaluated. Finally, a LC-HRMS profile and a molecular network of TDV extract were created to characterize the metabolites in the fruits' ethanolic extract.ResultsBioautography-TLC-HRMS followed by isolation and confirmation of the structure of the active compound by 1D and 2D NMR allowed the identification solamargine as the main compound responsible for the anti-Trichophyton mentagrophytes (MIC = 64 μg mL−1) and anti-Candida albicans (MIC = 64 μg mL−1) activities. In addition, solamargine led to a significant reduction of about 20% of the Pseudomonas aeruginosa biofilm formation. This effect was observed at a very low concentration (1.6 μg mL−1) and remained fairly consistent regardless of the concentration. In addition, solamargine reduced pyocyanin production by about 20% at concentrations of 12.5 and 50 μg mL−1. Furthermore, the LC-HRMS profiling of TDV allowed us to annotate seven known compounds that were analyzed through a molecular network.ConclusionsSolamargine has been shown to be the most active compound against T. mentoagrophytes and C. albicans in vitro. In addition, our data show that this compound affects significantly P. aeruginosa pyocyanin production and biofilm formation in our conditions. Altogether, these results might explain the traditional use of S. mammosum fruits to treat a variety of fungal infections and respiratory disorders