$Integration of Microfractionation, qNMR and zebrafish screening for the in vivo bioassay-guided isolation and quantitative bioactivity analysis of natural products.$
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Integration of Microfractionation, qNMR and zebrafish screening for the in vivo bioassay-guided isolation and quantitative bioactivity analysis of natural products.

Natural products (NPs) are an attractive source of chemical diversity for small-molecule drug discovery. Several challenges nevertheless persist with respect to NP discovery, including the time and effort required for bioassay-guided isolation of bioactive NPs, and the limited biomedical relevance t...

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Autores principales:	Nadine Bohni, María Lorena Cordero-Maldonado, Jan Maes, Dany Siverio-Mota, Laurence Marcourt, Sebastian Munck, Appolinary R Kamuhabwa, Mainen J Moshi, Camila V Esguerra, Peter A M de Witte, Alexander D Crawford, Jean-Luc Wolfender
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/deda4bd99d214c558b36959d3956097d
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spelling	oai:doaj.org-article:deda4bd99d214c558b36959d3956097d2021-11-18T07:45:07ZIntegration of Microfractionation, qNMR and zebrafish screening for the in vivo bioassay-guided isolation and quantitative bioactivity analysis of natural products.1932-620310.1371/journal.pone.0064006https://doaj.org/article/deda4bd99d214c558b36959d3956097d2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23700445/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Natural products (NPs) are an attractive source of chemical diversity for small-molecule drug discovery. Several challenges nevertheless persist with respect to NP discovery, including the time and effort required for bioassay-guided isolation of bioactive NPs, and the limited biomedical relevance to date of in vitro bioassays used in this context. With regard to bioassays, zebrafish have recently emerged as an effective model system for chemical biology, allowing in vivo high-content screens that are compatible with microgram amounts of compound. For the deconvolution of the complex extracts into their individual constituents, recent progress has been achieved on several fronts as analytical techniques now enable the rapid microfractionation of extracts, and microflow NMR methods have developed to the point of allowing the identification of microgram amounts of NPs. Here we combine advanced analytical methods with high-content screening in zebrafish to create an integrated platform for microgram-scale, in vivo NP discovery. We use this platform for the bioassay-guided fractionation of an East African medicinal plant, Rhynchosia viscosa, resulting in the identification of both known and novel isoflavone derivatives with anti-angiogenic and anti-inflammatory activity. Quantitative microflow NMR is used both to determine the structure of bioactive compounds and to quantify them for direct dose-response experiments at the microgram scale. The key advantages of this approach are (1) the microgram scale at which both biological and analytical experiments can be performed, (2) the speed and the rationality of the bioassay-guided fractionation - generic for NP extracts of diverse origin - that requires only limited sample-specific optimization and (3) the use of microflow NMR for quantification, enabling the identification and dose-response experiments with only tens of micrograms of each compound. This study demonstrates that a complete in vivo bioassay-guided fractionation can be performed with only 20 mg of NP extract within a few days.Nadine BohniMaría Lorena Cordero-MaldonadoJan MaesDany Siverio-MotaLaurence MarcourtSebastian MunckAppolinary R KamuhabwaMainen J MoshiCamila V EsguerraPeter A M de WitteAlexander D CrawfordJean-Luc WolfenderPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 5, p e64006 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Nadine Bohni María Lorena Cordero-Maldonado Jan Maes Dany Siverio-Mota Laurence Marcourt Sebastian Munck Appolinary R Kamuhabwa Mainen J Moshi Camila V Esguerra Peter A M de Witte Alexander D Crawford Jean-Luc Wolfender Integration of Microfractionation, qNMR and zebrafish screening for the in vivo bioassay-guided isolation and quantitative bioactivity analysis of natural products.
description	Natural products (NPs) are an attractive source of chemical diversity for small-molecule drug discovery. Several challenges nevertheless persist with respect to NP discovery, including the time and effort required for bioassay-guided isolation of bioactive NPs, and the limited biomedical relevance to date of in vitro bioassays used in this context. With regard to bioassays, zebrafish have recently emerged as an effective model system for chemical biology, allowing in vivo high-content screens that are compatible with microgram amounts of compound. For the deconvolution of the complex extracts into their individual constituents, recent progress has been achieved on several fronts as analytical techniques now enable the rapid microfractionation of extracts, and microflow NMR methods have developed to the point of allowing the identification of microgram amounts of NPs. Here we combine advanced analytical methods with high-content screening in zebrafish to create an integrated platform for microgram-scale, in vivo NP discovery. We use this platform for the bioassay-guided fractionation of an East African medicinal plant, Rhynchosia viscosa, resulting in the identification of both known and novel isoflavone derivatives with anti-angiogenic and anti-inflammatory activity. Quantitative microflow NMR is used both to determine the structure of bioactive compounds and to quantify them for direct dose-response experiments at the microgram scale. The key advantages of this approach are (1) the microgram scale at which both biological and analytical experiments can be performed, (2) the speed and the rationality of the bioassay-guided fractionation - generic for NP extracts of diverse origin - that requires only limited sample-specific optimization and (3) the use of microflow NMR for quantification, enabling the identification and dose-response experiments with only tens of micrograms of each compound. This study demonstrates that a complete in vivo bioassay-guided fractionation can be performed with only 20 mg of NP extract within a few days.
format	article
author	Nadine Bohni María Lorena Cordero-Maldonado Jan Maes Dany Siverio-Mota Laurence Marcourt Sebastian Munck Appolinary R Kamuhabwa Mainen J Moshi Camila V Esguerra Peter A M de Witte Alexander D Crawford Jean-Luc Wolfender
author_facet	Nadine Bohni María Lorena Cordero-Maldonado Jan Maes Dany Siverio-Mota Laurence Marcourt Sebastian Munck Appolinary R Kamuhabwa Mainen J Moshi Camila V Esguerra Peter A M de Witte Alexander D Crawford Jean-Luc Wolfender
author_sort	Nadine Bohni
title	Integration of Microfractionation, qNMR and zebrafish screening for the in vivo bioassay-guided isolation and quantitative bioactivity analysis of natural products.
title_short	Integration of Microfractionation, qNMR and zebrafish screening for the in vivo bioassay-guided isolation and quantitative bioactivity analysis of natural products.
title_full	Integration of Microfractionation, qNMR and zebrafish screening for the in vivo bioassay-guided isolation and quantitative bioactivity analysis of natural products.
title_fullStr	Integration of Microfractionation, qNMR and zebrafish screening for the in vivo bioassay-guided isolation and quantitative bioactivity analysis of natural products.
title_full_unstemmed	Integration of Microfractionation, qNMR and zebrafish screening for the in vivo bioassay-guided isolation and quantitative bioactivity analysis of natural products.
title_sort	integration of microfractionation, qnmr and zebrafish screening for the in vivo bioassay-guided isolation and quantitative bioactivity analysis of natural products.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/deda4bd99d214c558b36959d3956097d
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Integration of Microfractionation, qNMR and zebrafish screening for the in vivo bioassay-guided isolation and quantitative bioactivity analysis of natural products.

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