New psychoactive substances (NPS) have added a new dimension to the recreational drug scene. Pharmacological research is key in understanding and mitigating the harms associated with NPS. Previous research has shown that many NPS are more potent than conventional drugs in their ability to activate the different targets (whether receptors or transporters). However, detailed insight into the molecular mechanisms behind these effects are lagging behind. This project therefore puts a spotlight on the action of NPS via previously underexplored paradigms in pharmacology. Events both upstream and downstream of the molecular targets will be scrutinized, involving the use of in-house developed, as well as commercially available, bio-assays. The in vivo translatability will be assessed via in vivo assessments and by coupling the in vitro findings to forensic cases. Overall, this project aims at unraveling the harm potential of NPS by pulling back the curtain on their complex pharmacology.