(-)-11-Hydroxy-7'-isothiocyanato-1',1'- dimethylheptyl-.DELTA.8-THC: A Novel, High-Affinity Irreversible Probe for the Cannabinoid Receptor in the Brain

Guo, Yan; Abadji, Vasiliki; Morse, Kelley L.; Fournier, Donna J.; Li, Xiuyan; Makriyannis, Alexandros

doi:10.1021/jm00049a002

Cited by 39 publications

(50 citation statements)

References 2 publications

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“…We have reported the development of cannabinergic covalent probes and their potential application in GPCR binding site structural studies (Charalambous et al, 1992;Guo et al, 1994;Morse et al, 1995;Picone et al, 2002). The work described here clearly supports the role of the CWXP motif, which is found in Rhodopsin as well as other family 1A GPCRs, located in TMH6 of the CB1 receptor and its involvement in ligand recognition and binding.…”

supporting

confidence: 69%

“…This moiety will selectively react with more reactive nucleophiles, such as the thiol group of cysteine, but not with the hydroxyl of serine side chains or water. Preliminary studies with AM841 (R. P. Picone, D. J. Fournier, and A. Makriyannis, unpublished observations), as well as our earlier generations of C-3 side-chain functionalized isothiocyanates, demonstrated significant efficacy at irreversibly occupying CB1 binding sites in rat brain synaptosomes (Guo et al, 1994;Morse et al, 1995;Picone et al, 2002). This binding was not recoverable with sedimentation washing, which is widely accepted as evidence of irreversible occupation with nonradiolabeled spontaneously reacting isothiocyanate ligands (Guo et al, 1994;Morse et al, 1995;Zhu et al, 1996;Picone et al, 2002;Tahtaoui et al, 2003).…”

Section: Discussionmentioning

confidence: 91%

“…7). Unlike AM841, the above two ligands, in which the electrophilic isothiocyanate group is substituted for a -Br or -H, respectively, behave as fully reversible, wash-sensitive ligands indicative of their inability to irreversibly interact with CB1 (Guo et al, 1994;Picone et al, 2002;Tahtaoui et al, 2003).…”

Section: Resultsmentioning

confidence: 99%

“…One of our most successful designs encompasses a classical cannabinoid scaffold functionalized in the terminal position of the C-3 alkyl side chain (Charalambous et al, 1992;Guo et al, 1994;Morse et al, 1995;Picone et al, 2002). Together, these covalent probes have been very effective in irreversibly and selectively binding to CB1 in a highly specific manner.…”

Section: Discussionmentioning

confidence: 99%

“…Affinity Labeling. Labeling methods were adapted from our previously published procedures with a few modifications (Guo et al, 1994;Morse et al, 1995). Membranes containing CB1 receptor were allowed to equilibrate with AM841 for 1 h in a 30°C shaking water bath, after which suspensions were rapidly sedimented (27,200g) at 30°C.…”

Section: Methodsmentioning

confidence: 99%

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(-)-7′-Isothiocyanato-11-hydroxy-1′,1′-dimethylheptylhexahydrocannabinol (AM841), a High-Affinity Electrophilic Ligand, Interacts Covalently with a Cysteine in Helix Six and Activates the CB1 Cannabinoid Receptor

Picone

Khanolkar²,

Xu³

et al. 2005

Mol Pharmacol

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174

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The CB1 cannabinoid receptor has been shown to play important physiological roles in the central nervous system, as well as peripherally, and is a target for development of therapeutic medications. To gain insight on the ligand binding site(s) and structural features of activation, we designed and synthesized (Ϫ)-7Ј-isothiocyanato-11-hydroxy-1Ј,1Ј-dimethylheptylhexahydrocannabinol (AM841), a classical cannabinoid affinity label that incorporates an isothiocyanate substituent as an electrophilic reactive group capable of interacting irreversibly with a suitably located and properly oriented nucleophilic amino acid residue at or near the binding site. To obtain evidence for the site of covalent attachment of AM841, C6.47, identified in part by interactive ligand docking, was mutated to serine, alanine, and leucine to reduce or eliminate the nucleophilic character. Wild-type (WT) and mutant CB1 receptors were evaluated for their abilities to recognize a series of cannabinergic ligands. Each bound comparably to WT, excluding C6.47L, which displayed a reduced affinity for It is noteworthy that AM841 was shown to bind irreversibly to WT CB1 but exhibited no covalent attachment with the mutants and behaved as an agonist suggesting irreversible attachment to C6.47 maintains CB1 in its active state. The evidence presented identifies C6.47 as the site of covalent bond formation with AM841 and combined with the binding data fully supports the molecular modeling. These studies present the first report of tandem applications of affinity labeling, site-directed mutagenesis, and interactive ligand docking for CB1.The CB1 and CB2 cannabinoid receptors are relatively new members in the G-protein-coupled receptor (GPCR) superfamily. They have been shown to play important physiological roles and represent targets for development of therapeutic medications. From a pharmacological standpoint, agonist activation of both receptors results in the release of G␣ iproteins, causing a concomitant reduction in intracellular This work has been supported by National Institute on Drug Abuse grants DA05955 (to R.P.P.) DA00355 (to A.D.K.), DA09158, DA03801, DA07215, DA07312 (to A.M.), DA03934, DA00489 (to P.H.R.), DA05274, and DA09978 (to M.E.A.).

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supporting

confidence: 69%

Section: Discussionmentioning

confidence: 91%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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(-)-7′-Isothiocyanato-11-hydroxy-1′,1′-dimethylheptylhexahydrocannabinol (AM841), a High-Affinity Electrophilic Ligand, Interacts Covalently with a Cysteine in Helix Six and Activates the CB1 Cannabinoid Receptor

Picone

Khanolkar²,

Xu³

et al. 2005

Mol Pharmacol

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174

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Functionalized Cannabinoid Subtype 2 Receptor Ligands: Fluorescent, PET, Photochromic and Covalent Molecular Probes

2020

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Cannabinoid subtype 2 receptors (CB 2 Rs) are G protein‐coupled receptors (GPCRs) belonging to the endocannabinoid system, a complex network of signalling pathways leading to the regulation of key physiological processes. Interestingly, CB 2 Rs are strongly up‐regulated in pathological conditions correlated with the onset of inflammatory events like cancer and neurodegenerative diseases. Therefore, CB 2 Rs represent an important biological target for therapeutic as well as diagnostic purposes. No CB 2 R‐selective drugs are yet on the market, thus underlining a that deeper comprehension of CB 2 Rs’ complex activation pathways and their role in the regulation of diseases is needed. Herein, we report an overview of pharmacological and imaging tools such as fluorescent, positron emission tomography (PET), photochromic and covalent selective CB 2 R ligands. These molecular probes can be used in vitro as well as in vivo to investigate and explore the unravelled role(s) of CB 2 Rs, and they can help to design suitable CB 2 R‐targeted drugs.

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Design and Synthesis of Highly Potent and Specific ABHD6 Inhibitors

et al. 2021

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Fine‐tuning than complete disruption of 2‐arachidonoylglycerol (2‐AG) metabolism in the brain represents a promising pharmacological approach to limit potential untoward effects associated with complete blockade of monoacylglycerol lipase (MGL), the primary hydrolase of 2‐AG. This could be achieved through a/b‐hydrolase domain containing 6 (ABHD6) inhibition, which will provide a smaller and safer contribution to 2‐AG regulation in the brain. Pharmacological studies with ABHD6 inhibitors have recently been reported, where modulation of ABHD6 activity either through CB1R‐dependent or CB1R‐independent processes showed promise in preclinical models of epilepsy, neuropathic pain and inflammation. Furthermore in the periphery, ABHD6 modulates 2‐AG and other fatty acid monoacylglycerols (MAGs) and is implicated in Type‐2 diabetes, metabolic syndrome and potentially other diseases. Herein, we report the discovery of single‐digit nanomolar potent and highly specific ABHD6 inhibitors with >1000‐fold selectivity against MGL and FAAH. The new ABHD6 inhibitors provide early leads to develop therapeutics for neuroprotection and the treatment of inflammation and diabetes.

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(-)-11-Hydroxy-7'-isothiocyanato-1',1'- dimethylheptyl-.DELTA.8-THC: A Novel, High-Affinity Irreversible Probe for the Cannabinoid Receptor in the Brain

Cited by 39 publications

References 2 publications

(-)-7′-Isothiocyanato-11-hydroxy-1′,1′-dimethylheptylhexahydrocannabinol (AM841), a High-Affinity Electrophilic Ligand, Interacts Covalently with a Cysteine in Helix Six and Activates the CB1 Cannabinoid Receptor

(-)-7′-Isothiocyanato-11-hydroxy-1′,1′-dimethylheptylhexahydrocannabinol (AM841), a High-Affinity Electrophilic Ligand, Interacts Covalently with a Cysteine in Helix Six and Activates the CB1 Cannabinoid Receptor

Functionalized Cannabinoid Subtype 2 Receptor Ligands: Fluorescent, PET, Photochromic and Covalent Molecular Probes

Design and Synthesis of Highly Potent and Specific ABHD6 Inhibitors

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