Ricerc@Sapienza

The expected outcome of the project is to acquire a deeper knowledge of the most recent and concerning NPS. The proposed approach will allow on the one hand of reinforcing the existing monitoring instruments and methods facilitating the detection of NPS and their metabolites in seizures and biological samples; on the other hand, new data about NPS pharmaco-toxicology and biotransformation, will be collected and provided to the competent agencies (national early warning system of DPA, EMCDDA, Law Enforcement) for supporting forensic laboratories and Legal Medicine activity.
From an analytical point of view an outcome of the project will be the creation of a public database that will facilitate the work of forensic analysts exploiting existing platforms such as GNPS for untargeted MS raw data analysis and library search. MN is likely to improve the detectability of unexpected NPS through connection of MS/MS spectra with known and related molecules, additional algorithms that will be explored will make it possible to find additional connections, even when the overall fragmentation behaviour is altered, as long as there is some overlap in fragmentation pattern. The use of MN for NPS analysis represents a paradigm shift, opening new perspectives to identify unknown structurally related NPS by networking to MS/MS spectral data of existing related molecules. This innovative approach will be exploited along the project to possibly identify new molecules in seizures collected in the Italian territory by law enforcement, new drugs not included among the reference spectra can be putatively annotated as a result of the proposed activities.
By reinforcing the existing monitoring instruments and methods, the project will facilitate the detection of NPS and their metabolites in seizures and biological samples.
The proposed action will have an important impact in the fight against new psychoactive substances in Italy and in Europe. In fact, the development of new tools for detection of NPS by LC-HRMS will support the analytical and forensic researchers in the detection of NPS in seizures and biological samples. The national forensic laboratories constitute the eyes of the European EWS on new psychoactive compounds. The creation of a new database will promote communication among these laboratories and the use of innovative informatic tools will assist in the annotation of unknown NPS. Concurrently to the acquisition of new MS/MS data, accessible suspect lists and libraries will be leveraged to create a publicly available tandem mass spectral NPS database in order to address issues related to data inhomogeneity and limited access of existing resources. Collaboration and exchange of data between different institutes will support the actions on new drugs and will strengthen the capabilities of NEWS in the fight against NPS. It will represent a pivotal tool for the management of the NPS crisis.
On the other hand, the collection of new data about NPS metabolism and toxicokinetic is among the priorities of the Department for Anti-Drug Policies (DPA) of the Presidency of the Council of Ministers, providing a specialized support on the identification of NPS to preserve public health, and taking part to the National Early Warning System (SNAP). The development, management and amendment of effective drug policies relies heavily on the availability of accurate and timely information on the drug situation. In vivo preclinical evaluation of NPS in mice will allows to identify their main metabolites to be used as markers in toxicological and forensic research in humans. Alerting hospitals of the identities on NPS being used and predicting changes for clinical treatment needs based on longer-term monitoring are but two potential examples.
Another innovation of the present research consists in the use of metabolomics to identify potential biomarkers that can act as indirect indicators of drug consumption. The metabolomic study will provide further insights into the pharmaco-toxicological effects of the NPS under investigation. This approach attempts to understand how different drug classes affect the endogenous metabolites enabling delineation of altered biochemical pathways associated with NPS consumption and revealing disrupted molecular mechanisms. This approach promises to be interesting to tackle the problem of the increasing number of new drugs flooding the market. At present, only few studies have been performed, nevertheless showing promising first results to identify analytical biomarkers by metabolomics-related techniques. However, far more studies will be necessary for a final conclusion on the general suitability of metabolomics in drug testing, which will need also a dedicated data mining, such as advanced multivariate analysis.