Zebrafish Bioassay-guided Microfractionation for the Rapid in vivo Identification of Pharmacologically Active Natural Products
Soura Challal,
Nadine Bohni,
Olivia E. Buenafe,
Camila V. Esguerra,
Peter A. M. de Witte,
Jean-Luc Wolfender,
Alexander D. Crawford
Affiliations
Soura Challal
SCS-DSM Award for best poster presentation, Phytochimie et Produits Naturels Bioactifs Ecole de Pharmacie Genève-Lausanne Section des Sciences Pharmaceutiques Université de Genève Quai Ernest-Ansermet 30 CH-1211 Genève 4, Switzerland
Nadine Bohni
Phytochimie et Produits Naturels Bioactifs Ecole de Pharmacie Genève-Lausanne Section des Sciences Pharmaceutiques Université de Genève Quai Ernest-Ansermet 30 CH-1211 Genève 4, Switzerland
Olivia E. Buenafe
Laboratory for Molecular Biodiscovery Department of Pharmaceutical & Pharmacological Sciences University of Leuven Herestraat 49, P.O. Box 824 BE-3000 Leuven, Belgium; Department of Chemistry Ateneo de Manila University,
Manila, Philippines
Camila V. Esguerra
Laboratory for Molecular Biodiscovery Department of Pharmaceutical & Pharmacological Sciences University of Leuven Herestraat 49, P.O. Box 824 BE-3000 Leuven, Belgium
Peter A. M. de Witte
Laboratory for Molecular Biodiscovery Department of Pharmaceutical & Pharmacological Sciences University of Leuven Herestraat 49, P.O. Box 824 BE-3000 Leuven, Belgium
Jean-Luc Wolfender
Phytochimie et Produits Naturels Bioactifs Ecole de Pharmacie Genève-Lausanne Section des Sciences Pharmaceutiques Université de Genève Quai Ernest-Ansermet 30 CH-1211 Genève 4, Switzerland. [email protected]
Alexander D. Crawford
Laboratory for Molecular Biodiscovery Department of Pharmaceutical & Pharmacological Sciences University of Leuven Herestraat 49, P.O. Box 824 BE-3000 Leuven, Belgium. [email protected]
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.
