Notes: This review delves into the drug discovery pipeline, focusing on both phenotypic drug discovery (PDD) and target-based drug discovery (TDD) and the importance of experiments which determine the mechanism of action (MOA) of drugs. NanoBRET is highlighted as a highly specific assay for the measurement of target interaction. Specifically, the precise tagging of a target greatly minimizes assay cross-talk. (5122)

Notes: Cyclin G associated kinase (GAK) is a central regulator of viral and bacterial host cell entry. The synthesis and characterization of selective and potent GAK inhibitors is described. Inhibitors were assessed for ability to compete for GAK binding using the NanoBRET Target Engagement system. Briefly, a BRET signal was observed with the NanoLuc-GAK fusion in the presence of a red-shifted dye specific to the GAK ATP binding site. Upon treatment with GAK inhibitors, the dye was competed away, and a loss of BRET signal was observed. (5130)

Notes: This review mentions, under kinome-wide techniques, the NanoBRET™ assay for measuring the cellular target engagement across 178 members of a kinome using a technique involving competition between a compound of interest and a reversible cell-permeable energy transfer probe for binding to NanoLuc-tagged kinase in live cells. The authors note that, with the assay's fluorescent readout, it is amenable to higher throughput than mass spec-based techniques. (4964)

Notes: The researchers demonstrate use of the NanoBRET™ Target Engagement Intracellular Kinase Assays to report on kinase target engagement in real time for quantitative inhibitor profiling of 178 kinases, including over 40 integral membrane receptors. (4936)