Single-position ligand modifications tune CB2R activity by targeting the toggle switch

TL;DR

Researchers modified a single position of a ligand to control CB2R activity by targeting a key receptor residue, enabling diverse functional outcomes.

Contribution

A new strategy for modulating CB2R activity by targeting the toggle switch through single-position ligand modifications is introduced.

Findings

Modifications at a single ligand position modulate CB2R toggle switch Trp2586.48 to control receptor activity.

Ligands exhibit a range of functional outcomes, including full agonism and partial inverse agonism.

Compound (S)-1 shows biased signaling and unique pharmacological behavior due to toggle switch interactions.

Abstract

Cannabinoid receptor type 2 (CB2R) is a prominent class A G protein-coupled receptor (GPCR) and is a therapeutic target of interest for inflammatory diseases, pain management, and neurodegenerative disorders. We report the development of ligands based on HU-308 that share a single central scaffold but bear diverse sidechains, enabling controlled modulation of GPCR activation. Structural modifications at a single position of the parent ligand allow modulation of the single-residue toggle switch of CB2R, Trp2586.48, and thereby control over receptor activity. A continuum of functional outcomes is achieved through interaction of the ligands with the CB2R toggle switch, leading to full agonism, partial agonism, neutral antagonism, or partial inverse agonism. Several low-efficacy ligands display protean behavior across assays, underscoring context-dependent modulation of CB2R and its importance in profiling such ligands. A notable compound within this series is CF3-substituted (S)-1, which displays distinct CB2R affinity, potency, and a biased CB2R signaling profile. We provide a rationale based on molecular dynamics simulations for the unique pharmacological profile observed and suggest that stabilization of an active receptor conformation occurs by close-contact interaction of (S)-1 with the CB2R toggle switch. Our findings demonstrate that strategic structural modifications of class A GPCR ligands may, by targeting a receptor's toggle switch, shift ligands to different positions along the efficacy spectrum, independent of their parent scaffold's original functional profile.

This study, exemplified at CB2R, showcases how strategic reutilization of a single ligand scaffold fine-tunes activity of a class A GPCR by virtue of targeting its toggle switch.

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