Development and validation of an ADRβ1-SNAP-tag/CMC model for screening ADRβ1 antagonists: Application to nebivolol and its byproducts

Heart failure is a global epidemic characterized by high mortality and morbidity rates. β1-Adrenoceptors (ADRβ1) antagonists are the main drugs used in the treatment of HF. A simple and convenient method for screening ADRβ1 antagonists is necessary. Nebivolol has four chiral centers, and its byproducts, which are not easy to avoid, may interact with ADRβ1. Cell membrane chromatography based on SNAP-tag technology provides a comprehensive solution. In this study, an ADRβ1 target cell membrane chromatography model based on SNAP-tag technology was established, and the selectivity, specificity, repeatability, and column life of the system were validated. Under optimized conditions, this model was used to screen for active components targeting ADRβ1 in nebivolol and its byproducts. The bioactivity of nebivolol and its byproducts was detected by calcium influx. The results showed that the ADRβ1 SNAP-tag/CMC model was successfully established and could be used to screen active drugs targeting ADRβ1. Among nebivolol and its byproducts, D-nebivolol (SRRR-enantiomer) had the best affinity for ADRβ1. Three byproducts of nebivolol also exhibited retention characteristics on ADRβ1. In vitro calcium influx experiments verified the interaction of these components with ADRβ1, showing good inhibitory effects on ADRβ1. In conclusion, the ADRβ1 SNAP-tag/CMC model developed in this study can be used for the comprehensive screening of active components targeting ADRβ1 in complex samples.

Cell membrane chromatography; Nebivolol; SNAP-tag technology; β1-Adrenoceptors.