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Signalling bias in new drug discovery: detection, quantification and therapeutic impact

Abstract

Agonists of seven-transmembrane receptors, also known as G protein-coupled receptors (GPCRs), do not uniformly activate all cellular signalling pathways linked to a given seven-transmembrane receptor (a phenomenon termed ligand or agonist bias); this discovery has changed how high-throughput screens are designed and how lead compounds are optimized for therapeutic activity. The ability to experimentally detect ligand bias has necessitated the development of methods for quantifying agonist bias in a way that can be used to guide structure–activity studies and the selection of drug candidates. Here, we provide a viewpoint on which methods are appropriate for quantifying bias, based on knowledge of how cellular and intracellular signalling proteins control the conformation of seven-transmembrane receptors. We also discuss possible predictions of how biased molecules may perform in vivo, and what potential therapeutic advantages they may provide.

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Figure 1: Trafficking of receptor stimulus by agonists.
Figure 2: Bias plots.
Figure 3: Chemokine activation of CCR5.
Figure 4: Logistical scheme for quantifying signalling bias for agonists in a system-independent manner.
Figure 5: Interplay of tissue sensitivity and ligand bias.
Figure 6: The interaction of the receptor and allosteric vector with cellular signalling components.
Figure 7: The impact of signalling bias on drug screening.
Figure 8: Biased allosteric agonism and modulation.

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Acknowledgements

Work in A.C.'s laboratory is supported by the Program Grant No. 519461 and Project Grant No. APP1026962 of the National Health and Medical Research Council (NHMRC) of Australia, and Discovery Grant No. DP110100687 of the Australian Research Council. A.C. is an NHMRC Principal Research Fellow.

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Correspondence to Terry Kenakin.

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Competing interests

A.C. has a research contract with Servier (France), and recent consultancies with Johnson and Johnson (USA) and XOMA (USA). He is a Scientific Advisory Board member for Audeo, Australia. T.K. declares no competing financial interests.

Supplementary information

Supplementary information S1 (box)

The Black–Leff Operational Model for Modelling Drug Response (PDF 201 kb)

Supplementary information S2 (box)

Differences in Operational (Functional) Affinity (PDF 277 kb)

Supplementary information S3 (box)

Conditional Affinity of Allosteric Systems: 7 Transmembrane Receptors (PDF 232 kb)

Supplementary information S4 (box)

Fitting the Operational Model (PDF 209 kb)

Supplementary information S5 (box)

Equiactive Concentration Comparison for Pathways: Method of Barlow, Scott and Stephenson (PDF 197 kb)

Supplementary information S6 (box)

The Molecular Determinants of Efficacy within the Allosteric Vector (PDF 220 kb)

Supplementary information S7 (box)

Equating ΔΔLog(τ/KA) and βlig values (PDF 246 kb)

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Glossary

[35S]GTPγS

35S-labelled GTP; a non-hydrolysable G protein-activating analogue of GTP that is used to measure interactions between seven-transmembrane receptors (also known as GPCRs) and G proteins.

Allosteric binding site

The site on a seven-transmembrane receptor protein (also known as GPCR) to which modulators bind to affect the subsequent binding and effect of another ligand that interacts with the receptor; this ligand is usually the endogenous agonist binding to its cognate (that is, orthosteric) binding site.

Conditional affinity

The measured affinity of a ligand for a seven-transmembrane receptor (also known as GPCR) when the receptor is bound to an allosteric guest molecule (such as a G protein or β-arrestin). The conditional affinity of the ligand for the receptor will vary with the concentration and type of the guest molecule that is co-bound.

Efficacy

The property of a molecule that causes a change in the behaviour of a seven-transmembrane receptor (also known as GPCR) towards the cell when the molecule is bound to the receptor.

EC50

The concentration of an agonist that produces 50% of the maximal response to the agonist for a defined signalling response pathway.

Full agonists

Agonists that induce the maximum obtainable response that can be produced by a signalling system.

Operational affinity

Also referred to as functional affinity. The apparent equilibrium dissociation constant of the agonist–receptor complex, as determined by fitting the Black–Leff operational model to agonist concentration–response curves.

Orthosteric agonist

An agonist that binds to the same binding site on the seven-transmembrane receptor protein (also known as GPCR) as the endogenous agonist (that is, the orthosteric binding site).

pEC50

Negative logarithm of the EC50 (the concentration of an agonist that produces 50% of the maximal response to the agonist for a defined signalling response pathway).

Receptor coupling efficiencies

A term that describes the degree of seven-transmembrane receptor (also known as GPCR) occupancy by an agonist; receptor coupling efficiency relates to the resulting cellular response. A low receptor occupancy that yields a large cellular response constitutes a high coupling efficiency.

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Kenakin, T., Christopoulos, A. Signalling bias in new drug discovery: detection, quantification and therapeutic impact. Nat Rev Drug Discov 12, 205–216 (2013). https://doi.org/10.1038/nrd3954

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