Characterization of the intrinsic activity for a novel class of cannabinoid receptor ligands: Indole quinuclidine analogs

Franks, Lirit N.; Ford, Benjamin M.; Madadi, Nikhil Reddy; Penthala, Narsimha Reddy; Crooks, Peter A.; Prather, Paul L.

doi:10.1016/j.ejphar.2014.05.007

Cited by 14 publications

(16 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…IQD analogues PNR-4-20 and PNR-4-02 were synthesized as previously described (30) and were dissolved in 100% DMSO to 10 mM stock concentrations for in vitro assays. WIN-55,212-2, JWH-018, CP-55,940, and DAMGO were obtained from Tocris Bioscience (Bristol, United Kingdom).…”

Section: 0 Methodsmentioning

confidence: 99%

“…Membranes were prepared as described previously (30). Briefly, cell pellets from CHO-β2-hCB 1 , CHO-hCB 1 -Rx and CHO-hMOR cells were homogenized separately with 10 complete strokes utilizing a 40 mL Dounce glass homogenizer in ice-cold buffer containing 3 mM MgCl 2 , 50 mM HEPES pH 7.4, and 1 mM EDTA.…”

Section: 0 Methodsmentioning

confidence: 99%

“…The next day, culture media was gently removed from each well and replaced with 500 µl pre-incubation mixture containing either HAM’s F-12 K (CHO-β2-hCB 1 ) or DMEM (CHO-hMOR) with 2 μCi/well [ 3 H]adenine, 0.9% NaCl, and 500 µM 3-isobutyl-1-methylxathine (IBMX), as previously reported (30). Cells were subsequently incubated for 3 hr at 37°C.…”

Section: 0 Methodsmentioning

confidence: 99%

“…Although a number of structurally diverse classes of cannabinoid ligands have been developed (25–27), CB 1 R agonists characterized to date exhibit relatively equivalent coupling to both G-protein and β-arrestin 2 pathways (28). Our group recently characterized a novel class of indole quinulidinone (IQD) analogues that exhibit high nanomolar affinity for CB 1 Rs (29), and act as partial to full CB 1 R agonists for modulation of the Gi/Go-coupled intracellular effector adenylyl cyclase (30). Although IQD analogues have been shown to clearly couple CB 1 Rs to G protein-dependent pathways, the ability of compounds in this class to produce CB 1 R-mediated G protein-independent recruitment of β-arrestin 2 has yet to be determined.…”

Section: 0 Introductionmentioning

confidence: 99%

“…Analogues based on the novel IQD structural scaffold (30) might bias coupling of CB 1 Rs towards either G protein or β-arrestin 2 signaling pathways. Therefore, the purpose of this study was to determine whether cannabinoid agonists in this class exhibit ligand bias at CB 1 Rs.…”

Section: 0 Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Characterization of structurally novel G protein biased CB 1 agonists: Implications for drug development

Ford

Franks

Tai

et al. 2017

Pharmacological Research

Self Cite

View full text Add to dashboard Cite

The human cannabinoid subtype 1 receptor (hCB1R) is highly expressed in the CNS and serves as a therapeutic target for endogenous ligands as well as plant-derived and synthetic cannabinoids. Unfortunately, acute use of hCB1R agonists produces unwanted psychotropic effects and chronic administration results in development of tolerance and dependence, limiting the potential clinical use of these ligands. Studies in β-arrestin knockout mice suggest that interaction of certain GPCRs, including μ-, δ-, κ-opioid and hCB1Rs, with β-arrestins might be responsible for several adverse effects produced by agonists acting at these receptors. Indeed, agonists that bias opioid receptor activation toward G-protein, relative to β-arrestin signaling, produce less severe adverse effects. These observations indicate that therapeutic utility of agonists acting at hCB1Rs might be improved by development of G-protein biased hCB1R agonists. Our laboratory recently reported a novel class of indole quinulidinone (IQD) compounds that bind cannabinoid receptors with relatively high affinity and act with varying efficacy. The purpose of this study was to determine whether agonists in this novel cannabinoid class exhibit ligand bias at hCB1 receptors. Our studies found that a novel IQD-derived hCB1 receptor agonist PNR-4-20 elicits robust G protein-dependent signaling, with transduction ratios similar to the non-biased hCB1R agonist CP-55,940. In marked contrast to CP-55,940, PNR-4-20 produces little to no β-arrestin 2 recruitment. Quantitative calculation of bias factors indicates that PNR-4-20 exhibits from 5.4-fold to 29.5-fold bias for G protein, relative to β-arrestin 2 signaling (when compared to G protein activation or inhibition of forskolin-stimulated cAMP accumulation, respectively). Importantly, as expected due to reduced β-arrestin 2 recruitment, chronic exposure of cells to PNR-4-20 results in significantly less desensitization and down-regulation of hCB1Rs compared to similar treatment with CP-55,940. PNR-4-20 (i.p.) is active in the cannabinoid tetrad in mice and chronic treatment results in development of less persistent tolerance and no significant withdrawal signs when compared to animals repeatedly exposed to the non-biased full agoinst JWH-018 or Δ9-THC. Finally, studies of a structurally similar analog PNR-4-02 show that it is also a G protein biased hCB1R agonist. It is predicted that cannabinoid agonists that bias hCB1R activation toward G protein, relative to β-arrestin 2 signaling, will produce fewer and less severe adverse effects both acutely and chronically.

show abstract

Section: 0 Methodsmentioning

confidence: 99%

Section: 0 Methodsmentioning

confidence: 99%

Section: 0 Methodsmentioning

confidence: 99%