5'-Azido-.DELTA.8-THC: a novel photoaffinity label for the cannabinoid receptor

Charalambous, Avgui; Guo, Yan; Houston, Devin B.; Howlett, A­llyn C.; Compton, David R.; Martin, Billy R.; Makriyannis, Alexandros

doi:10.1021/jm00094a023

Cited by 39 publications

(51 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: 70%

“…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%

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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: 70%

Section: Discussionmentioning

confidence: 99%

(-)-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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“…10,12,24 Thus, (-)-5'-hydroxyl-∆ 8 -tetrahydrocannabinol 18 was converted to the triflic diester 19 (58% yield) through reaction with triflic anhydride in the presence of 2,6-lutidine. Because of its tendency to decompose, 19 was purified by flash column chromatography and was immediately used in the next step.…”

Section: Synthesismentioning

confidence: 99%

“…To a solution of 18 10,12,24 (303 mg, 0.92 mmol) and 2,6-lutidine (0.53 mL, 4.58 mmol) in methylene chloride (6 mL) cooled to 0°C was added triflic anhydride (0.68 mL, 4.06 mmol), and the mixture was stirred for 45 minutes. The solvent was then removed using a stream of nitrogen, and the residue obtained was purified by flash column chromatography on florisil using 5% diethyl ether in hexane to yield 319 mg (58%) of fairly pure (- …”

Section: (-)-5'-fluoro-∆ ∆ 8 -Tetrahydrocannabinol (20)mentioning

confidence: 99%

The role of halogen substitution in classical cannabinoids: A CB1 pharmacophore model

Nikas

Grzybowska

Papahatjis

et al. 2004

AAPS J

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The presence of halogens within the classical cannabinoid structure leads to large variations in the compounds' potencies and affinities for the CB1 receptors. To explore the structure activity relationships within this class of analogs we have used a series of halogen-substituted (-)-∆ 8 -tetrahydrocannabinol analogs and compared their affinities for the CB1 cannabinoid receptor. Our results indicate that halogen substitution at the end-carbon of the side chain leads to an enhancement in affinity with the bulkier halogens (Br, I) producing the largest effects. Conversely, 2-iodo substitution on the phenolic ring leads to a 2-fold reduction in affinity while iodo-substitution in the C1'-position of the side chain lowers the compound's affinity for CB1 by more than 8-fold. The pharmacophoric requirements resulting from halogen-substitution are explored using computer modeling methods.

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“…Earlier studies had demonstrated that a wide range of small substituents (hydroxy, bromo, cyano, azido, acetamido, etc. ) could be incorporated into the terminus of the side chain (Charalambous et al, 1992;Martin et al, 1993Martin et al, , 1999. Importantly, these studies demonstrated that the nitrogencontaining substituents increased CB 1 receptor affinity and pharmacological potency.…”

mentioning

confidence: 99%

Pharmacological Characterization of Novel Water-Soluble Cannabinoids

Martin

Wiley²,

Beletskaya³

et al. 2006

J Pharmacol Exp Ther

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Presently, there are numerous structural classes of cannabinoid receptor agonists, all of which require solubilization for experimental purposes. One strategy for solubilizing water-insoluble tetrahydrocannabinols is conversion of the phenolic hydroxyl to a morpholinobutyryloxy substituent. The hydrochloride salts of these analogs are water-soluble and active in vivo when administered in saline. The present investigation demonstrated that hydrochloride salts of numerous substituted butyryloxy esters are water-soluble and highly potent. The substitutions include piperidine, piperazine, and alkyl-substituted amino moieties. It was also discovered that incorporation of a nitrogenous moiety in the alkyl side chain increased the pharmacological potency of tetrahydrocannabinol. For example, an analog containing a pyrazole in the side chain (O-2545) was found to have high affinity and efficacy at cannabinoid 1 (CB 1 ) and CB 2 receptors, and when dissolved in saline, it was highly efficacious when administered either intravenously or intracerebroventricularly to mice. A series of carboxamido and carboxylic acid amide analogs exhibited high pharmacological potency, but their hydrochloride salts were not water-soluble. On the other hand, incorporation of imidazoles into the terminus of the side chain led to water-soluble hydrochloride salts that were highly potent when administered in saline to laboratory animals. It is now possible to conduct cannabinoid research with agonists that are water-soluble and thus obviating the need of solubilizing agents.Marijuana has attracted considerable attention for centuries because of its psychotropic and medicinal properties. Early scientific investigations were conducted with either smoked plant material or the plant extract. Needless to say, the synthesis of marijuana's major psychotropic constituent, ⌬ 9 -THC, opened a new era in marijuana research (Gaoni and Mechoulam, 1964). For the first time researchers were able to conduct research in a quantitative fashion because the precise dose of ⌬ 9 -THC could be administered. Unfortunately, ⌬ 9 -THC is a noncrystalline, highly lipophilic compound that requires solubilization with either a surfactant agent or adherence to a water miscible substance (albumin, Tween 80, Emulphor, etc.). This high lipophilicity has placed constraints on the pharmacological evaluation of ⌬ 9 -THC. Even when dissolved in these vehicles, ⌬ 9 -THC has limited solubility and will precipitate if care is not exercised. This cannabinoid will adhere to solid surfaces rather than remain in solution under certain conditions. There is always the concern that the use of different vehicles in separate pharmacological studies may influence the pharmacological effects of ⌬ 9 -THC. Undoubtedly, these challenges in drug solubilization contribute to the vagaries accompanying cannabinoid data collection. Although many of the above vehicles are sufficient for systemic administration of cannabinoids, they present significant challenges for cannabinoid administration int...

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5'-Azido-.DELTA.8-THC: a novel photoaffinity label for the cannabinoid receptor

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

The role of halogen substitution in classical cannabinoids: A CB1 pharmacophore model

Pharmacological Characterization of Novel Water-Soluble Cannabinoids

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