Bioassays to study biased signalling of novel synthetic opioids

摘要: Objective For hundreds of years, opioids have been used in the management pain, mediating their analgesic effect via binding to Mu opioid receptor (MOR). Fentanyl and morphine are examples MOR agonists whose effects associated with unwanted side such as tolerance dependence. While analgesia is mainly realized through G protein-signalling pathway, undesirable linked β-arrestin pathway. Little known about a potential “bias” synthetic – including fentanyl analogs – that emerged on illegal drug market. To develop novel, robust platform study activity both protein pathways. Methods We developed bioassays based functional complementation two split fragments Nanoluciferase (LgBiT SmBiT), either fused or cytosolic proteins: mini β-arrestin. These assays were performed 96-well plates using transiently transfected HEK293 T cells, pure reference standards. Results Assays set up which MOR, LgBiT, was combined proteins, SmBiT. Both demonstrated concentration-dependent responses at hydromorphone, DAMGO, carfentanil, activation low 10−13M still being detectable for carfentanil. A wide range analogs currently tested–the results will be discussed. Discussion The present provides an vitro by NPS means capacity couple recruit Conclusion not only allow insight into signalling opioids, may help better understand why certain higher toxicity but also screen biofluids activity.