New Horizons on Molecular Pharmacology Applied to Drug Discovery: When Resonance Overcomes Radioligand Binding

作者：Pernomian Larissa; Gomes Mayara Santos; Moreira Josimar Dornelas; Tomich de Paula da Silva Carlos Henrique; Campos Rosa Joaquin Maria; de Barros Cardoso Cristina Ribeiro

来源：Current Radiopharmaceuticals, 2017, 10(1): 16-20.

DOI：10.2174/1874471010666170208152420

<jats:sec>
<jats:title />
<jats:p>One of the cornerstones of rational drug development is the measurement of molecular parameters
derived from ligand-receptor interaction, which guides therapeutic windows definition.
Over the last decades, radioligand binding has provided valuable contributions in this field as key
method for such purposes. However, its limitations spurred the development of more exquisite techniques
for determining such parameters. For instance, safety risks related to radioactivity waste, expensive
and controlled disposal of radioisotopes, radiotracer separation-dependence for affinity
analysis, and one-site mathematical models-based fitting of data make radioligand binding a suboptimal
approach in providing measures of actual affinity conformations from ligands and G proteincoupled
receptors (GPCR). Current advances on high-throughput screening (HTS) assays have
markedly extended the options of sparing sensitive ways for monitoring ligand affinity. The advent
of the novel bioluminescent donor NanoLuc luciferase (Nluc), engineered from Oplophorus gracilirostris
luciferase, allowed fitting bioluminescence resonance energy transfer (BRET) for monitoring
ligand binding. Such novel approach named Nluc-based BRET (NanoBRET) binding assay consists
of a real-time homogeneous proximity assay that overcomes radioligand binding limitations but ensures
the quality in affinity measurements. Here, we cover the main advantages of NanoBRET protocol
and the undesirable drawbacks of radioligand binding as molecular methods that span pharmacological
toolbox applied to Drug Discovery. Also, we provide a novel perspective for the application
of NanoBRET technology in affinity assays for multiple-state binding mechanisms involving
oligomerization and/or functional biased selectivity. This new angle was proposed based on specific
biophysical criteria required for the real-time homogeneity assigned to the proximity NanoBRET
protocol.</jats:p>
</jats:sec>

• 出版日期2017