Small molecules discovery from marine organisms and microorganisms: A new pipeline combing LC-HRMS and NMR metabolomics

TASCMAR is an EU funded project that aspires to develop new tools and strategies to overcome existing bottlenecks in the bio-discovery and industrial exploitation of novel marine derived bio-molecules with applications in pharmaceuticals, nutraceuticals and cosmeceuticals as anti-aging ingredients. An integrated, holistic technological approach including NMR- and MS-based metabolomics has been applied, in conjunction with bioactivity profiling, as a filtering and a bio-prioritization tool for the selection of the top 10% most promising extracts. In the frame of the project, more than 180 organisms (53% Demospongiae, 11% Holothuroidea, 10% Anthozoa, 7.5% Echinoidea, 4% Gastropoda, others < 3%) collected from the Indian Ocean, the Red Sea and Mediterranean, were extracted using DCM/MeOH. All extracts have been analysed by a strategy combining UHPLC/Orbitrap-HRMS and NMR (600 MHz). Sample profiles were acquired following an optimized standard procedure and statistical evaluation of the results was performed by multivariate analysis (MVA), e.g. PCA, PLS-DA, and OPLS. Statistical total correlation spectroscopy (STOCSY) analysis was implemented for the discovery of correlated and anticorrelated spectral features responsible for the clustering observed. On the level of class both non-supervised (PCA) and supervised analysis (PLS-DA) showed good separation. On the level of order only the supervised analysis resulted in clustering formation. On the level of genus and species classification was observed only in certain cases, e.g., in the case of Biemna fortis or Clathria reinwardti samples collected from different spots were closely related, while in the case of Iotrochota baculifera samples even from the same spot exhibit high variation. Overall we consider UHPLC-HRMS and NMR metabolomics approach in combination with bioactivity profiling as a powerful tool for the prioritization of hits in large high throughput screening drug discovery projects.

Acknowledgements: This work has been financially supported by EU under the frame of TASCMAR project (H2020-BG-03 – 2014, Grant agreement: 634674).

Keywords: Anti-aging, marine invertebrates, marine microorganisms, metabolomics.