Phase-Trafficking Methods in Natural Products, Modulators of Organic Anion Transporting Polypeptides from Rollinia emarginata, and Pregnane and Cardiac Glycosides from Asclepias spp.

For decades, chemists and medicinal chemists have found in nature the source of inspiration for drug discovery and development. This work describes several aspects of the interaction between the fields of natural products and medicinal chemistry, from isolation and characterization of bioactive molecules to semi-synthetic analogs preparation. A new phase-trafficking approach for acidic, basic, and neutral compounds separation from organic plant extracts was developed, validated and successfully applied to crude plant extracts. This new method could be applied to natural extracts of diverse origin in order to generate better quality samples for initial bioassays. Furthermore, this new catch-and-release methodology allowed the isolation and identification of three compounds new to the literature from the extensively studied ginger rhizomes. Using a more traditional bioassay guided fractionation, we have identified six small-molecules from Rollinia emarginata that modulate organic anion transporting polypeptide´s (OATPs) function. The results of this study show that diverse plant materials are a promising source for the isolation of OATP modulating compounds, and that a bioassay-guided approach can be used to efficiently identify selective OATP modulators. In addition, a 1H NMR-based metabolomic approach was used as a dereplication tool to study the effect of aqueous green tea extracts on OATP1B1-mediated uptake of estrone-3-sulfate. Our findings suggested that not only the gallate catechins were important for the observed uptake inhibition, but also compounds theogalline and 3-p-cumaroyl quinic acid could have been involved. A screening against breast cancer cell line Hs578T was conducted with ten plant species from the Asclepiadaceae family and, based on our findings, three plants were selected for detailed investigation: Asclepias verticillata, Asclepias syriaca, and Asclepias sullivantii. As a result, a total of 46 compounds were isolated and identified, half of which represented novel structures. The isolates showed a wide variety of structures including pregnane and cardiac glycosides, pentacyclic triterpenes, glycosylated flavonoids and lignans, among others. Furthermore, a group of cardiac glycosides were found to have strong cytotoxicity selected breast cancer cell lines. Finally, using a semi-synthetic approach, cardiac glycoside analogs with modifications in the butenolide ring were pursued in order to better understand their SAR. Starting from the commercially available trans-aldosterone, the cardiac glycoside core was built up using a microwave-promoted allylic oxidation using SeO2 (Riley oxidation). In addition, a microwave-promoted Miyaura-Suzuki cross-coupling was utilized to obtain the desired 17β-aryl analogs

Isolation of modulators of Organic Anion Transporting Polypeptides (OATPs) from Rollinia emarginata Schlecht (Annonaceae)

Comparative Medicine - OneHealth and Comparative Medicine Poster SessionOrganic Anion Transporting Polypeptides (OATPs) comprise a superfamily of sodium-independent membrane transporters which are involved in transporting numerous endogenous and exogenous substances. OATPs are expressed in different tissues such as intestine, liver, kidney and brain, and are responsible for the uptake of important drugs including cholesterol-lowering agents (statins), endothelin receptor antagonists (sartans), the anticancer drugs methotrexate, SN-38, paclitaxel and docetaxel, as well as the antibiotic rifampicin. Through a strategic collaboration, we search for novel small molecules from the organic extract of Rollinia emarginata Schlecht. (Annonaceae) that interact with the liver specific OATP1B1 and OATP1B3 applying a bioassay guided isolation approach. The organic extract was fractionated using different chromatographic techniques, and each fraction was tested for its effect on OATP1B1- and OATP1B3-mediated transport of 1µM estrone-3-sulfate and 0.1µM estradiol-17||-glucuronide. Several inhibitors, including both substrate-specific and non-specific, were isolated and chemically identified. For instance, the compound Quercetin 3-O-||-L-arabinopyranosyl (1 ->2)||-L-rhamnopyranoside was shown to inhibit both OATP1B1- and OATP1B3-mediated transport of estradiol-17||-glucuronide by more than 90%, relative to control (DMSO). However, with respect to transport of 1µM estrone-3-sulfate it inhibits OATP1B1 by only 45% while, interestingly, stimulating transport mediated by OATP1B3 (2 fold over control). Thanks to our collaborative efforts, we were able to show that plants can be suitable source of small molecules that modulate OATPs using bioassay guided isolation approach

Application of Phase-Trafficking Methods to Natural Products Research

A novel simultaneous phase-trafficking approach using spatially separated solid-supported reagents for rapid separation of neutral, basic, and acidic compounds from organic plant extracts with minimum labor is reported. Acidic and basic ion-exchange resins were physically separated into individual sacks (“tea bags”) for trapping basic and acidic compounds, respectively, leaving behind in solution neutral components of the natural mixtures. Trapped compounds were then recovered from solid phase by appropriate suspension in acidic or basic solutions. The feasibility of the proposed separation protocol was demonstrated and optimized with an “artificial mixture” of model compounds. In addition, the utility of this methodology was illustrated with the successful separation of the alkaloid skytanthine from Skytanthus acutus Meyen and the main catechins and caffeine from Camellia sinensis L. (Kuntze). This novel approach offers multiple advantages over traditional extraction methods, as it is not labor intensive, makes use of only small quantities of solvents, produces fractions in adequate quantities for biological assays, and can be easily adapted to field conditions for bioprospecting activities

Plan Estratégico de Mercadeo para el Restaurante Verde Fusión

Trabajo Final de Graduación (Maestría en Administración de Empresas) Instituto Tecnológico de Costa Rica, Escuela de Administración de Empresas, 2019.En los últimos años nuestro país se encuentra experimentando dos fenómenos: uno de ellos en el ámbito de la salud y la alimentación saludable y el segundo es el auge de los servicios. Cada día las personas se preocupan más por mejorar la calidad de sus vidas mediante la alimentación y el ejercicio, pero se topan con una limitante que es el tiempo que le pueden dedicar a la preparación de los alimentos debido a todas las obligaciones laborales, académicas y personales. Los estilos de vida cada vez son más ajetreados y las personas buscan sacar el máximo provecho de su tiempo. El negocio de Restaurante Verde Fusión consiste en ofrecer el servicio de venta de comida saludable y personalizada según los requerimientos de cada consumidor para todas las personas que están interesadas en cuidar su estilo de vida y que disfrutan de una dieta saludable y balanceada. Los clientes podrán realizar sus pedidos en línea por los diferentes canales de comunicación y sus productos serán entregados de forma puntual donde ellos lo deseen. Todo esto como una solución muy conveniente ante la problemática de la limitación de tiempo y de la búsqueda de una mejora en la calidad de vida por medio de la alimentación saludable. Los ingresos serán provenientes de los pedidos en línea, se desarrollará una página web con la opción de “agregar al carrito” y se habilitarán canales como WhatsApp Empresarial, Facebook, Instagram, así como también el desarrollo de una aplicación para dispositivos móviles la cual será uno de los valores agregados y diferenciadores con respecto a los competidores. Los clientes serán capaces de realizar sus pedidos, agregar productos al carrito de compras y se habilitara una opción de “subir su plan nutricional”, esta opción para los clientes que desean utilizar el servicio de comida personalizada. Los pedidos se entregarán 2 veces por semana con el fin de conservar el sabor, la frescura y la calidad de los alimentos. Este plan estratégico de mercadeo tiene como objetivo colaborar al crecimiento de un emprendimiento que consideramos que tiene mucho potencial de mercado ya que las nuevas tendencias así lo comprueban, Paez (2018) sostiene que “En temas de nutrición, el año nos está dejando tendencias muy interesantes como: la dieta cetogénica, alimentación limpia, ayuno intermitente, probióticos y “meal prep” entre otros”. Por otro lado, Abissi (2018) sostiene que “El patrón de consumo de los hogares en Costa Rica ha cambiado y ahora, gastan más en servicios como Internet, telefonía, servicios de comida y bebida y menos en bienes como alimentos, prendas de vestir, equipos eléctricos y otros como refrigeradoras, cocinas y artículos para el hogar”. Estas dos citas son un claro ejemplo de cómo una buena combinación y estrategia entre comida saludable y servicios personalizados que den un valor agregado son un excelente negocio para aquellos empresarios que tengan los recursos para invertir en este nuevo segmento de mercado emergente.In recent years Costa Rica has experienced two phenomena: a significant increase in health and wellness awareness and people new lifestyles. People are learning the ability to take action towards optimal health, to create the proper environment for a healthier lifestyle and of course the type of food they eat, but they very often encounter the same dilemma, food-related time in order to prepare their own food and their interests and personal activities. For many people, finding time to exercise, carry on with their activities and prepare healthy meals become very challenging because of the time constraints. Verde Fusion sells healthy food and personalized healthy food options. Customers will be able to order the products on Verde Fusion’s social media channels and will be reach via WhatsApp. The payment method is via wire transfer and location for delivery is required. These new services are providing solutions to an unattended market who has been facing struggling with this situation. We are aiming for Verde Fusion to implement an online food ordering approach with the main goal of centralized all the foods orders and provide pleasant shopping experience to the customers. Clients will be able to place their orders, add products to the shopping cart and enable an option to "upload your nutritional plan", this option is for customers who want to use the personalized food service. Orders will be delivered twice per week in order to preserve food properties and taste. WhatsApp Business will continue for customer services purposes, Facebook, Instagram will be enhanced and enable to provide more information to all the followers and new followers and customers.This plan aims to increase sales, increase revenue growth and implement a customer’s retention and loyalty program. Verde Fusion has the potential to gain in the market according to the studies done and feedback collected

A tea catechin, epigallocatechin-3-gallate, is a unique modulator of the farnesoid X receptor

Farnesoid X receptor (FXR) is a ligand-activated nuclear receptor and serves as a key regulator to maintain health of the liver and intestine. Bile acids are endogenous ligands of FXR, and there are increasing efforts to identify FXR modulators to serve as biological probes and/or pharmaceutical agents. Natural FXR ligands isolated from plants may serve as models to synthesize novel FXR modulators. In this study, we demonstrated that epigallocatechin-3-gallate (EGCG), a major tea catechin, specifically and dose-dependently activates FXR. In addition, EGCG induced FXR target gene expression in vitro. Surprisingly, in a co-activator (SRC2) recruitment assay, we found that EGCG does not recruit SRC2 to FXR, but it dose-dependently inhibits recruitment of SRC2 to FXR (IC50, 1 μM) by GW6064, which is a potent FXR synthetic ligand. In addition, EGCG suppressed FXR target gene expression induced by either GW4064 or chenodeoxycholic acid in vitro. Furthermore, wild-type and FXR knockout mice treated with an acute dose of EGCG had induced mRNA expression in a subset of FXR target genes in the intestine but not in the liver. In conclusion, EGCG is a unique modulator of FXR in the intestine and may serve as an important model for future development of FXR modulators

Hunters of the Puna de Salta on a broad chronological scale (ca. 11,000-3,000 BP): hunting archaeological landscape in the sector of Pozo Cavado, Pocitos basin

Se presenta nueva evidencia para caracterizar el paisaje arqueológico cinegético del sector de Pozo Cavado, cuencade Pocitos, Puna de Salta, entre ca. 11000 y 3000 años AP. En particular, se analizan los conjuntos arquitectónicos ylíticos. En las construcciones, se reconocen parapetos y muros extensos que podrían haber conformado un espaciode caza intensa de camélidos, mientras que entre el material lítico, se relevaron los artefactos tiempo-sensitivos,principalmente vinculados con tecnología extractiva. Específicamente, se recuperó una preforma de punta cola depescado en un contexto superficial, cuyo hallazgo representa información relevante debido a la escasez de esta clase deartefactos en el Noroeste argentino. También se distinguieron puntas triangulares apedunculadas tempranas, aunquepredominan distintas variantes lanceoladas. Estas últimas, en conjunto con el registro cronológico y arqueológico encapa del sitio Abrigo Pozo Cavado, indican una mayor intensidad de ocupación a lo largo del Holoceno medio y, enparticular, hacia fines de este período. Más generalmente, el conjunto de los indicadores analizados parece señalarla construcción de un paisaje de caza perdurable en una escala cronológica de largo plazo.New evidence is presented to characterize the archaeological hunting landscape of the Pozo Cavado sector, Pocitos basin, Puna de Salta, between ca. 11,000 and 3,000 years BP. In particular, the architectural and lithic ensembles are analyzed. In the constructions, parapets and walls are recognized, which could have formed a space for intense hunting of camelids, while among the lithic material, time-sensitive artifacts, mainly linked to extractive technology, were studied. Specifically, a fishtail point preform was recovered in a superficial context, which finding represents relevant information due to the scarcity of this class of artifacts in the Northwestern Argentina. Early triangular nonstemmed points were also found, although different lanceolate variants predominate. These morphologies, together with the chronological and archaeological record in layer of the Abrigo Pozo Cavado site, indicate a greater intensity of human occupations throughout the middle Holocene and, in particular, towards the end of this period. More generally, the whole of evidence seem to point to the construction of an enduring hunting landscape on a long-term chronological scale.Fil: Lopez, Gabriel Eduardo Jose. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Orsi, Juan Pablo. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Seguí, Silvina Tatiana. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Araya, Sonia Mariel. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; ArgentinaFil: Sola, Patricia. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Coloca, Federico Ignacio. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Pre-hispanic human occupations in the basins of Pastos Grandes, Pocitos and Ratones, Puna de Salta, Argentina: contributions to the archaeological study from diverse lines of evidence

Se presenta el análisis de distintas líneas de evidencia con el objetivo de caracterizar las ocupaciones humanas prehispánicas en las cuencas de Pastos Grandes, Pocitos y Ratones, Puna de Salta. A partir de la generación de un corpus de información arqueológica en escala regional, se busca discutir los procesos de cambio ocurridos a lo largo del Holoceno entre ca. 10.000 AP y ca. 500 AP. Esta escala cronológica y espacial amplia permite analizar la variabilidad de procesos desarrollados desde las primeras ocupaciones humanas en la región hasta la expansión Inca. Los materiales analizados provienen de los sitios Alero Cuevas, Abrigo Pozo Cavado, Abra de Minas y Cueva Inca Viejo. Las líneas de evidencia corresponden a diversos conjuntos arqueofaunísticos, arqueobotánicos, líticos y cerámicos. Los resultados de esta investigación fueron relevantes para estudiar procesos tales como la intensificación y la domesticación de camélidos, los cambios en la movilidad y en los mecanismos de interacción, y el impacto Inca en la región, entre otros.We analysed different lines of evidence in order to characterize pre-Hispanic human occupations in the basins of Pastos Grandes, Pocitos and Ratones, Puna de Salta, Argentina. The aim is to discuss, based on a corpus of regional archaeological information, the change processes that occurred throughout the Holocene between ca. 10,000 BP and ca. 500 BP. This wide chronological and spatial scale allows us to analyze the variability of processes developed from the first human occupations in the region to the Inca expansion. The materials studied come from the archaeological sites called Alero Cuevas, Abrigo Pozo Cavado, Abra de Minas and Cueva Inca Viejo. The lines of evidence correspond to archaeofaunal, archaeobotanical, lithic and ceramic sets. The results of this research were relevant to study processes such as the intensification and domestication of camelids, changes in mobility and interaction mechanisms, and the Inca impact in the region, among others.Fil: Lopez, Gabriel Eduardo Jose. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; ArgentinaFil: Araya, Sonia Mariel. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; ArgentinaFil: Seguí, Silvina Tatiana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; ArgentinaFil: Orsi, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; ArgentinaFil: Coloca, Federico Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; ArgentinaFil: Sola, Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; Argentin

Hunters of the Puna de Salta on a broad chronological scale (ca. 11,000-3,000 BP): hunting archaeological landscape in the sector of Pozo Cavado, Pocitos basin

Se presenta nueva evidencia para caracterizar el paisaje arqueológico cinegético del sector de Pozo Cavado, cuencade Pocitos, Puna de Salta, entre ca. 11000 y 3000 años AP. En particular, se analizan los conjuntos arquitectónicos ylíticos. En las construcciones, se reconocen parapetos y muros extensos que podrían haber conformado un espaciode caza intensa de camélidos, mientras que entre el material lítico, se relevaron los artefactos tiempo-sensitivos,principalmente vinculados con tecnología extractiva. Específicamente, se recuperó una preforma de punta cola depescado en un contexto superficial, cuyo hallazgo representa información relevante debido a la escasez de esta clase deartefactos en el Noroeste argentino. También se distinguieron puntas triangulares apedunculadas tempranas, aunquepredominan distintas variantes lanceoladas. Estas últimas, en conjunto con el registro cronológico y arqueológico encapa del sitio Abrigo Pozo Cavado, indican una mayor intensidad de ocupación a lo largo del Holoceno medio y, enparticular, hacia fines de este período. Más generalmente, el conjunto de los indicadores analizados parece señalarla construcción de un paisaje de caza perdurable en una escala cronológica de largo plazo.New evidence is presented to characterize the archaeological hunting landscape of the Pozo Cavado sector, Pocitos basin, Puna de Salta, between ca. 11,000 and 3,000 years BP. In particular, the architectural and lithic ensembles are analyzed. In the constructions, parapets and walls are recognized, which could have formed a space for intense hunting of camelids, while among the lithic material, time-sensitive artifacts, mainly linked to extractive technology, were studied. Specifically, a fishtail point preform was recovered in a superficial context, which finding represents relevant information due to the scarcity of this class of artifacts in the Northwestern Argentina. Early triangular nonstemmed points were also found, although different lanceolate variants predominate. These morphologies, together with the chronological and archaeological record in layer of the Abrigo Pozo Cavado site, indicate a greater intensity of human occupations throughout the middle Holocene and, in particular, towards the end of this period. More generally, the whole of evidence seem to point to the construction of an enduring hunting landscape on a long-term chronological scale.Fil: Lopez, Gabriel Eduardo Jose. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Orsi, Juan Pablo. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Seguí, Silvina Tatiana. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Araya, Sonia Mariel. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; ArgentinaFil: Sola, Patricia. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Coloca, Federico Ignacio. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Arqueología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Cytotoxic Oleanane-type Triterpenen Saponins with N-acetyl Sugar from Albizia inundata [abstract]

Comparative Medicine - OneHealth and Comparative Medicine Poster SessionBioassay-guided fractionation of a CH2Cl2-MeOH extract of the aerial part of Albizia inundata , collected in Argentina, resulted in the isolation of two new oleanane-type triterpene saponins with N-acetyl sugar (1, 2) and seven known saponins (3-9). Their structures were established on the basis of extensive 2D NMR (HSQC, COSY or DQF-COSY, TOCSY, HMBC), ESIMS and chemical methods. Saponins 1-4 showed cytotoxicity against the in JMAR, MDA 1986, UM-SCC-2 and JHU011 human head and neck squamous cells using the MTS assay