Miniaturization of the Structure Elucidation of Novel Natural Products - Two Trace Antibacterial Acylated Caprylic Alcohol Glycosides from<i>Arctostaphylos pumila</i>

Hu, Jin‐Feng; Yoo, Hye‐Dong; Williams, Caroline; Garo, Eliane; Cremin, Peadar; Zeng, Lu; Vervoort, Hélène C.; Lee, Chris M.; Hart, Shane M.; Goering, Matt G.; O'Neil–Johnson, Mark; Eldridge, Gary R.

doi:10.1055/s-2005-837787

Cited by 30 publications

(31 citation statements)

References 11 publications

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“…Depending on the potency of these isolated compounds, the requirement of only microgram amounts to induce an initial biological response represents another excellent benefit of using zebrafish as a model organism over other higher vertebrates (e.g. rodents, in which the active dose requirements are usually a thousand-fold higher [21]). …”

Section: Introductionmentioning

confidence: 99%

Integration of Microfractionation, qNMR and Zebrafish Screening for the In Vivo Bioassay-Guided Isolation and Quantitative Bioactivity Analysis of Natural Products

et al. 2013

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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.

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Section: Introductionmentioning

confidence: 99%

Integration of Microfractionation, qNMR and Zebrafish Screening for the In Vivo Bioassay-Guided Isolation and Quantitative Bioactivity Analysis of Natural Products

et al. 2013

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“…This was accomplished by using protocols similar to previous works performed by one of the authors (J.‐F. Hu) .…”

Section: Methodsmentioning

confidence: 99%

“…The natural product compound libraries screened in this study included 10 000 flash fractions and 50 000 high-performance liquid chromatography (HPLC) fractions that were generated from 671 different herbs/materials widely used in TCMs. This was accomplished by using protocols similar to previous works performed by one of the authors (J.-F. Hu) [21,22].…”

Section: Compound Librarymentioning

confidence: 99%

A novel glucagon‐like peptide 1 peptide identified from Ophisaurus harti

Zhu

Huang

Gao

et al. 2013

Journal of Peptide Science

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Glucagon-like peptide 1 receptor (GLP1R) is a promising target for the treatment of type 2 diabetes. Because of the short half-life of endogenous GLP1 peptide, other GLP1R agonists are considered to be appealing therapeutic candidates. A high-throughput assay has been established to screen for GLP1R agonists in a 60 000-well natural product compound library fractionated from 670 different herbs/materials widely used in traditional Chinese medicines (TCMs). The screening is based on primary screen of GLP1R⁺ reporter gene assay with the counter screen in GLP1R⁻ cell line. An active fraction, A089-147, was identified from the screening. Fraction A089-147 was isolated from dried Ophisaurus harti, and the fact that its GLP1R agonist activity was sensitive to trypsin treatment indicates its peptidic nature. The active ingredient of A089-147 was later identified as O. harti GLP1 through transcriptome analysis. Chemically synthesized O. harti GLP1 showed GLP1R agonist activity and sensitivity to dipeptidase IV digestion. This study illustrated a comprehensive screening strategy to identify novel GLP1R agonists from TCMs libraries and at the same time underlined the difficulty of identifying a non-peptidic GLP1R agonist. The novel O. harti GLP1 peptide yielded from this study confirmed broader application of TCMs libraries in active peptide identification.

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“…The same microfractionation procedure described for LC-SPE-NMR can be used for analyzing extracts; however, in this case, the LC peaks are not trapped on SPE cartridges but simply collected into vials or in 96-well plates and usually dried by speed vacuum evaporation. Each sample can be dissolved in a volume slightly exceeding the volume of the flow cell (6.5 mL for a 5 mL flow cell volume), filtered, injected into the probe, and parked in the center of the micro-coil by pushing it with an adjusted push volume (typically 8-10 mL) [76,77]. The disadvantage of at-line work is that more sample handling is needed compared to the on-flow approach.…”

Section: Spe-nmr Microflow Nmr and Nmr With Cryogenized Probesmentioning

confidence: 99%

Modern Approaches in the Search for New Active Compounds from Crude Extracts of Natural Sources

Queiroz

Hostettmann

Wolfender

2012

Plant Bioactives and Drug Discovery

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Miniaturization of the Structure Elucidation of Novel Natural Products - Two Trace Antibacterial Acylated Caprylic Alcohol Glycosides fromArctostaphylos pumila

Cited by 30 publications

References 11 publications

Integration of Microfractionation, qNMR and Zebrafish Screening for the In Vivo Bioassay-Guided Isolation and Quantitative Bioactivity Analysis of Natural Products

Integration of Microfractionation, qNMR and Zebrafish Screening for the In Vivo Bioassay-Guided Isolation and Quantitative Bioactivity Analysis of Natural Products

A novel glucagon‐like peptide 1 peptide identified from Ophisaurus harti

Modern Approaches in the Search for New Active Compounds from Crude Extracts of Natural Sources

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