Projects per year
Abstract
Background and Purpose The cannabinoid receptor type 1 (CB1) has an allosteric binding site. The drugs ORG27569 {5-chloro-3-ethyl-N-[2-[4-(1- piperidinyl)phenyl]ethyl]-1H-indole-2-carboxamide} and PSNCBAM-1 {1-(4-chlorophenyl)-3-[3-(6-pyrrolidin-1-ylpyridin-2-yl)phenyl]urea} have been extensively characterized with regard to their effects on signalling of the orthosteric ligand CP55,940 {(-)-cis-3-[2-hydroxy-4-(1,1-dimethylheptyl)phenyl]- trans-4-(3-hydroxypropyl)cyclohexanol}, and studies have suggested that these allosteric modulators increase binding affinity but act as non-competitive antagonists in functional assays. To gain a deeper understanding of allosteric modulation of CB1, we examined real-time signalling and trafficking responses of the receptor in the presence of allosteric modulators. Experimental Approach Studies of CB1 signalling were carried out in HEK 293 and AtT20 cells expressing haemagglutinin-tagged human and rat CB1. We measured real-time accumulation of cAMP, activation and desensitization of potassium channel-mediated cellular hyperpolarization and CB1 internalization. Key Results ORG27569 and PSNCBAM-1 produce a complex, concentration and time-dependent modulation of agonist-mediated regulation of cAMP levels, as well as an increased rate of desensitization of CB 1-mediated cellular hyperpolarization and a decrease in agonist-induced receptor internalization. Conclusions and Implications Contrary to previous studies characterizing allosteric modulators at CB1, this study suggests that the mechanism of action is not non-competitive antagonism of signalling, but rather that enhanced binding results in an increased rate of receptor desensitization and reduced internalization, which results in time-dependent modulation of cAMP signalling. The observed effect of the allosteric modulators is therefore dependent on the time frame over which the signalling response occurs. This finding may have important consequences for the potential therapeutic application of these compounds.
Original language | English |
---|---|
Pages (from-to) | 893-907 |
Number of pages | 15 |
Journal | British Journal of Pharmacology |
Volume | 170 |
Issue number | 4 |
DOIs | |
Publication status | Published - Oct 2013 |
Bibliographical note
Copyright the Publisher 2013. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.Keywords
- cannabinoid receptor
- allosteric modulator
- desensitization
- internalization
- receptor signalling
Fingerprint
Dive into the research topics of 'Real-time characterization of cannabinoid receptor 1 (CB1) allosteric modulators reveals novel mechanism of action'. Together they form a unique fingerprint.Projects
- 1 Finished
-
Molecular targets of amino acid/neurotransmitter conjugates of fatty acids
Connor, M., Vaughan, C., McIntyre, P., Kassiou, M. & Chinchen, G.
1/02/11 → 31/12/16
Project: Research