Zebrafish Bioassay-guided Microfractionation for the Rapid in vivo Identification of Pharmacologically Active Natural Products
- Authors
- Challal, S., Bohni, N., Buenafe, O.E., Esguerra, C.V., de Witte, P.A., Wolfender, J.L., and Crawford, A.D.
- ID
- ZDB-PUB-120529-19
- Date
- 2012
- Source
- Chimia 66(4): 229-232 (Journal)
- Registered Authors
- Buenafe, Olivia Erin, Crawford, Alexander, de Witte, Peter, Esguerra, Camila V.
- Keywords
- microflow NMR, microfractionation, natural products, UHPLC-TOF-MS, zebrafish
- MeSH Terms
-
- Animals
- Biological Assay/methods*
- Biological Products/chemistry*
- Biological Products/pharmacology
- Chromatography, High Pressure Liquid/methods
- Mass Spectrometry/methods
- Zebrafish
- PubMed
- 22613156 Full text @ Chimia (Aarau)
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.