Polypharmacological profiling across protein target families and cellular pathways using the multiplexed cell-based assay platform safetyProfiler reveals efficacy, potency and side effects of drugs

Selectivity profiling is key for assessing the pharmacological properties of multi-target drugs. We have developed a cell-based and barcoded assay encompassing ten druggable targets, including GPCRs, RTKs, nuclear receptors, a protease as well as their key downstream pathways and profiled 17 drugs in living cells for efficacy, potency, and side effects. Notably, this multiplex assay, termed safetyProfiler, enabled the simultaneous assessment of multiple target and pathway activities, shedding light on the polypharmacological profile of compounds. For example, the neuroleptics clozapine, paliperidone, and risperidone potently inhibited primary targets DRD2 and HTR2A as well as cAMP and calcium pathways. However, while paliperidone and risperidone also potently inhibited the secondary target ADRA1A and MAP kinase downstream pathways, clozapine only exhibited mild antagonistic effects on ADRA1A and lacked MAP kinase inhibition downstream of DRD2 and HTR2A. Our findings underscore the utility of such comprehensive profiling in elucidating the pharmacological properties of established therapeutics and facilitating the development of novel multi-target drugs with enhanced efficacy and selectivity.