title: Zebrafish bioassay-guided microfractionation for the rapid in vivo identification of pharmacologically active natural products.
abstract: The rapid acquisition of structural and bioactivity information on natural products (NPs) at the sub- milligram scale is key for performing efficient bioactivity-guided isolations. Zebrafish offer the possibility of rapid in vivo bioactivity analysis of small molecules at the microgram  scale - an attractive feature when combined with high-resolution fractionation technologies and analytical methods such as UHPLC-TOF-MS and microflow NMR. Numerous biomedically relevant assays are now available in zebrafish, encompassing most indication areas. Zebrafish also provide  the possibility to screen bioactive compounds for potential hepato-, cardio-, and neurotoxicities at a very early stage in the drug discovery process. Here we describe two strategies using zebrafish bioassays for the high-resolution in vivo bioactivity profiling of medicinal plants, using  either a one-step or a two-step procedure for active compound isolation directly into 96-well plates. The analysis of the microfractions by microflow NMR in combination with UHPLC-TOF-MS of the extract enables the rapid dereplication of compounds and an estimation of their microgram quantities  for zebrafish bioassays. Both the one-step and the two-step isolation procedures enable a rapid estimation of the bioactive potential of NPs directly from crude extracts. In summary, we present an in vivo , microgram-scale NP discovery platform combining zebrafish bioassays with microscale  analytics to identify, isolate and evaluate pharmacologically active NPs.
authors.affiliation: Université de Genève Quai Ernest-Ansermet, Genève 4, Switzerland.
authors.forename: Soura
authors.initials: S
authors.lastname: Challal
authors.forename: Nadine
authors.initials: N
authors.lastname: Bohni
authors.forename: Olivia E
authors.initials: OE
authors.lastname: Buenafe
authors.forename: Camila V
authors.initials: CV
authors.lastname: Esguerra
authors.forename: Peter A M
authors.initials: PA
authors.lastname: de Witte
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Wolfender
authors.forename: Alexander D
authors.initials: AD
authors.lastname: Crawford
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D001688:Biological Products / Q000737:chemistry* / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D015027:Zebrafish
chemical_list: D001688:Biological Products