Kinetic operational models of agonism for G-protein-coupled receptors

Hoare, Sam R.J.; Pierre, Nicolas; Moya, Arturo Gonzalez; Larson, Brad

doi:10.1016/j.jtbi.2018.02.014

Cited by 31 publications

(70 citation statements)

References 78 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, sometimes, it is difficult or impossible to estimate every kinetic micro-rate constant from typical functional assays for kinetic modelling (Hoare, Pierre, Moya, & Larson, 2018). Hence, there is a need to develop a reduced kinetic model that contains the essential FIGURE 6 Comparison of the potency/efficacy of CB 1 receptor agonists for internalisation in 3HA-hCB 1 HEK cells derived from model-free method (the inverse of mean residence time) and kinetic modelling approach (the quasi-steady state internalisation model).…”

Section: Figurementioning

confidence: 99%

Model‐free and kinetic modelling approaches for characterising non‐equilibrium pharmacological pathway activity: Internalisation of cannabinoid CB₁ receptors

Zhu

Finlay

Glass

et al. 2019

British J Pharmacology

View full text Add to dashboard Cite

Background and Purpose Receptor internalisation is by nature kinetic. Application of a standard equilibrium dose response model to describe the properties of a ligand inducing internalisation, while commonly used, are therefore problematic. Here, we propose two quantitative approaches to address this issue—(a) a model‐free method and (b) a kinetic modelling approach—and systematically evaluate the performance of these methods against traditional equilibrium methods to characterise the internalisation profiles of cannabinoid CB1 receptor agonists. Experimental Approach Kinetic internalisation assays were conducted using a concentration series of six CB1 receptor ligands. Internalisation rate analysis and snapshot equilibrium analysis were performed. A model‐free method was developed based on the mean residence time of internalisation. A kinetic internalisation model was developed under the quasi‐steady state assumption. Key Results Rates of receptor internalisation depended on both agonist and concentration. Agonist potencies from snapshot equilibrium analysis increased with stimulation time, and there was no single time point at which internalisation profiles could infer agonist properties in a comparative manner. The model‐free method yielded a time‐invariant measure of potency/efficacy for internalisation. The kinetic model adequately described the internalisation of CB1 receptors over time and provided robust estimates of both potency and efficacy. Conclusion and Implications Applying equilibrium analysis to a non‐equilibrium pathway cannot provide a reliable estimate of agonist potency. Both the model‐free and kinetic modelling approaches characterised the internalisation profiles of CB1 receptor agonists. The kinetic model provides additional advantages as a method to capture changes in receptor number during other functional assays.

show abstract

Section: Figurementioning

confidence: 99%

Model‐free and kinetic modelling approaches for characterising non‐equilibrium pharmacological pathway activity: Internalisation of cannabinoid CB₁ receptors

Zhu

Finlay

Glass

et al. 2019

British J Pharmacology

View full text Add to dashboard Cite

show abstract

“…A number of empirical drug parameters can be obtained from these data, such as the t1/2, plateau, AUC, or the signal at a single time point. An alternative and biologically meaningful parameter is the initial rate of signaling, analogous to the initial rate of enzyme activity 24,25 . We recently discovered this parameter can be easily obtainable from signaling kinetic data using a new kinetic pharmacological framework of GPCR signaling (Fig 3) 25 .…”

Section: Pharmacological Analysis Model Of Receptorarrestin Interactimentioning

confidence: 99%

“…This is the initial rate of signaling by the ligand-bound receptor and is termed kτ. This parameter can be easily estimated by curve fitting 24,25 .…”

Section: Pharmacological Analysis Model Of Receptorarrestin Interactimentioning

confidence: 99%

“…Regarding the data analysis, drug activity metrics are required which quantify the kinetics in terms that can be applied in establishing structure-activity relationships 23 . We recently developed a data analysis framework for G-protein and downstream signaling that quantifies kinetics in terms of the initial rate of signaling 24,25 . This rate, analogous to the initial rate of enzyme activity, is the rate of signaling before it is impacted by regulation of signaling mechanisms such as receptor desensitization and signal decay 25 .…”

Section: Introductionmentioning

confidence: 99%

“…This rate, analogous to the initial rate of enzyme activity, is the rate of signaling before it is impacted by regulation of signaling mechanisms such as receptor desensitization and signal decay 25 . This parameter, termed kτ, provides a biologically meaningful kinetic metric of ligand efficacy that has been applied to quantify ligand activity for G-protein activation and second-messenger generation 24,25 .…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

A new kinetic method for measuring agonist efficacy and ligand bias using high resolution biosensors and a kinetic data analysis framework

Hoare¹,

Tewson

Quinn

et al. 2019

Preprint

View full text Add to dashboard Cite

The kinetics/dynamics of signaling are of increasing value for G-protein-coupled receptor therapeutic development, including spatiotemporal signaling and the kinetic context of biased agonism. Effective application of signaling kinetics to developing new therapeutics requires reliable kinetic assays and an analysis framework to extract kinetic pharmacological parameters. Here we describe a platform for measuring arrestin recruitment kinetics to GPCRs using a high quantum yield, genetically encoded fluorescent biosensor, and a new analysis framework to quantify the recruitment kinetics. The sensor enabled high temporal resolution measurement of arrestin recruitment to the angiotensin AT1 and vasopressin V2 receptors. The analysis quantified the initial rate of arrestin signaling (kτ), a biologicallymeaningful kinetic drug efficacy parameter, by fitting time course data using routine curve-fitting methods. Biased agonism was assessed by comparing kτ values for arrestin recruitment with those for Gq signaling via the AT1 receptor. The kτ ratio values were in good agreement with bias estimates from existing methods. This platform potentially improves and simplifies assessment of biased agonism because the same assay modality is used to compare pathways (potentially in the same cells), the analysis method is parsimonious and intuitive, and kinetic context is factored into the bias measurement. Receptor excess over arrestin

show abstract

Luciferase Complementation Approaches to Measure GPCR Signaling Kinetics and Bias

Dijon

Nesheva

Holliday

2021

Methods in Molecular Biology

View full text Add to dashboard Cite

Kinetic operational models of agonism for G-protein-coupled receptors

Cited by 31 publications

References 78 publications

Model‐free and kinetic modelling approaches for characterising non‐equilibrium pharmacological pathway activity: Internalisation of cannabinoid CB₁ receptors

Model‐free and kinetic modelling approaches for characterising non‐equilibrium pharmacological pathway activity: Internalisation of cannabinoid CB₁ receptors

A new kinetic method for measuring agonist efficacy and ligand bias using high resolution biosensors and a kinetic data analysis framework

Luciferase Complementation Approaches to Measure GPCR Signaling Kinetics and Bias

Contact Info

Product

Resources

About

Kinetic operational models of agonism for G-protein-coupled receptors

Cited by 31 publications

References 78 publications

Model‐free and kinetic modelling approaches for characterising non‐equilibrium pharmacological pathway activity: Internalisation of cannabinoid CB1 receptors

Model‐free and kinetic modelling approaches for characterising non‐equilibrium pharmacological pathway activity: Internalisation of cannabinoid CB1 receptors

A new kinetic method for measuring agonist efficacy and ligand bias using high resolution biosensors and a kinetic data analysis framework

Luciferase Complementation Approaches to Measure GPCR Signaling Kinetics and Bias

Contact Info

Product

Resources

About

Model‐free and kinetic modelling approaches for characterising non‐equilibrium pharmacological pathway activity: Internalisation of cannabinoid CB₁ receptors

Model‐free and kinetic modelling approaches for characterising non‐equilibrium pharmacological pathway activity: Internalisation of cannabinoid CB₁ receptors