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Zebrafish bioassay-guided natural product discovery: isolation of angiogenesis inhibitors from East African medicinal plants.

Crawford AD, Liekens S, Kamuhabwa AR, Maes J, Munck S, Busson R, Rozenski J, Esguerra CV, de Witte PA - PLoS ONE (2011)

Bottom Line: Natural products represent a significant reservoir of unexplored chemical diversity for early-stage drug discovery.Zebrafish bioassay-guided fractionation identified the active components of these plants as emodin, an inhibitor of the protein kinase CK2, and coleon A lactone, a rare abietane diterpenoid with no previously described bioactivity.These results suggest that the combination of zebrafish bioassays with analytical chromatography methods is an effective strategy for the rapid identification of bioactive natural products.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Sciences, University of Leuven, Leuven, Belgium. alexander.crawford@pharm.kuleuven.be

ABSTRACT
Natural products represent a significant reservoir of unexplored chemical diversity for early-stage drug discovery. The identification of lead compounds of natural origin would benefit from therapeutically relevant bioassays capable of facilitating the isolation of bioactive molecules from multi-constituent extracts. Towards this end, we developed an in vivo bioassay-guided isolation approach for natural product discovery that combines bioactivity screening in zebrafish embryos with rapid fractionation by analytical thin-layer chromatography (TLC) and initial structural elucidation by high-resolution electrospray mass spectrometry (HRESIMS). Bioactivity screening of East African medicinal plant extracts using fli-1:EGFP transgenic zebrafish embryos identified Oxygonum sinuatum and Plectranthus barbatus as inhibiting vascular development. Zebrafish bioassay-guided fractionation identified the active components of these plants as emodin, an inhibitor of the protein kinase CK2, and coleon A lactone, a rare abietane diterpenoid with no previously described bioactivity. Both emodin and coleon A lactone inhibited mammalian endothelial cell proliferation, migration, and tube formation in vitro, as well as angiogenesis in the chick chorioallantoic membrane (CAM) assay. These results suggest that the combination of zebrafish bioassays with analytical chromatography methods is an effective strategy for the rapid identification of bioactive natural products.

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Crude methanolic extracts and isolated compounds exhibit concentration-dependent effects on vascular outgrowth in zebrafish embryos.Relative vascular outgrowth scores are shown (see Materials & Methods). a, O. sinuatum crude extract; b, P. barbatus crude extract; c, emodin; d, coleon AL.
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pone-0014694-g002: Crude methanolic extracts and isolated compounds exhibit concentration-dependent effects on vascular outgrowth in zebrafish embryos.Relative vascular outgrowth scores are shown (see Materials & Methods). a, O. sinuatum crude extract; b, P. barbatus crude extract; c, emodin; d, coleon AL.

Mentions: To test the utility of this zebrafish assay for natural product discovery, we screened crude methanolic extracts from over 80 East African medicinal plants. Two extracts, from Oxygonum sinuatum (Meisn.) Dammer (Polygonaceae) and Plectranthus barbatus Andrews (Lamiaceae), inhibited ISV outgrowth in fli-1:EGFP embryos in a dose-dependent manner (Figs. 1c, d and 2a, b).


Zebrafish bioassay-guided natural product discovery: isolation of angiogenesis inhibitors from East African medicinal plants.

Crawford AD, Liekens S, Kamuhabwa AR, Maes J, Munck S, Busson R, Rozenski J, Esguerra CV, de Witte PA - PLoS ONE (2011)

Crude methanolic extracts and isolated compounds exhibit concentration-dependent effects on vascular outgrowth in zebrafish embryos.Relative vascular outgrowth scores are shown (see Materials & Methods). a, O. sinuatum crude extract; b, P. barbatus crude extract; c, emodin; d, coleon AL.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3040759&req=5

pone-0014694-g002: Crude methanolic extracts and isolated compounds exhibit concentration-dependent effects on vascular outgrowth in zebrafish embryos.Relative vascular outgrowth scores are shown (see Materials & Methods). a, O. sinuatum crude extract; b, P. barbatus crude extract; c, emodin; d, coleon AL.
Mentions: To test the utility of this zebrafish assay for natural product discovery, we screened crude methanolic extracts from over 80 East African medicinal plants. Two extracts, from Oxygonum sinuatum (Meisn.) Dammer (Polygonaceae) and Plectranthus barbatus Andrews (Lamiaceae), inhibited ISV outgrowth in fli-1:EGFP embryos in a dose-dependent manner (Figs. 1c, d and 2a, b).

Bottom Line: Natural products represent a significant reservoir of unexplored chemical diversity for early-stage drug discovery.Zebrafish bioassay-guided fractionation identified the active components of these plants as emodin, an inhibitor of the protein kinase CK2, and coleon A lactone, a rare abietane diterpenoid with no previously described bioactivity.These results suggest that the combination of zebrafish bioassays with analytical chromatography methods is an effective strategy for the rapid identification of bioactive natural products.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Sciences, University of Leuven, Leuven, Belgium. alexander.crawford@pharm.kuleuven.be

ABSTRACT
Natural products represent a significant reservoir of unexplored chemical diversity for early-stage drug discovery. The identification of lead compounds of natural origin would benefit from therapeutically relevant bioassays capable of facilitating the isolation of bioactive molecules from multi-constituent extracts. Towards this end, we developed an in vivo bioassay-guided isolation approach for natural product discovery that combines bioactivity screening in zebrafish embryos with rapid fractionation by analytical thin-layer chromatography (TLC) and initial structural elucidation by high-resolution electrospray mass spectrometry (HRESIMS). Bioactivity screening of East African medicinal plant extracts using fli-1:EGFP transgenic zebrafish embryos identified Oxygonum sinuatum and Plectranthus barbatus as inhibiting vascular development. Zebrafish bioassay-guided fractionation identified the active components of these plants as emodin, an inhibitor of the protein kinase CK2, and coleon A lactone, a rare abietane diterpenoid with no previously described bioactivity. Both emodin and coleon A lactone inhibited mammalian endothelial cell proliferation, migration, and tube formation in vitro, as well as angiogenesis in the chick chorioallantoic membrane (CAM) assay. These results suggest that the combination of zebrafish bioassays with analytical chromatography methods is an effective strategy for the rapid identification of bioactive natural products.

Show MeSH
Related in: MedlinePlus