Classical Cannabinoids Research Articles

Structure-activity analysis of anandamide analogs: relationship to a cannabinoid pharmacophore.

Anandamides are endogenous fatty acid ethanolamides that have been shown to bind to the cannabinoid receptor and possess cannabimimetic activity yet are structurally dissimilar from the classical cannabinoids found in Cannabis sativa. We have employed molecular dynamics studies of a variety of anandamides to characterize their conformational mobility and determine whether there are pharmacophoric similarities with delta 9-THC. We have found that a looped conformation of these arachidonyl compounds is energetically favorable and that a structural correlation between this low-energy conformation and the classical cannabinoids can be obtained with the superposition of (1) the oxygen of the carboxyamide with the pyran oxygen in delta 9-THC, (2) the hydroxyl group of the ethanol with the phenolic hydroxyl group of delta 9-THC, (3) the five terminal carbons and the pentyl side chain of delta9-THC, and (4) the polyolefin loop overlaying with the cannabinoid tricyclic ring. The shape similarity is extended to show that other fatty acid ethanolamides that possess varying degrees of unsaturation also vary in their conformational mobility, which affects their ability to overlay with delta 9-THC as described above. Within this series of compounds, the most potent analog, the tetraene (arachidonyl) analog (i.e., anandamide itself), was determined to have restricted conformational mobility that favored an optimal pharmacophore overlay with delta9-THC. Eight pharmacologically active anandamide analogs are shown to have similar conformational mobility and pharmacophore alignments that are conformationally accessible. Furthermore, when these compounds are aligned to delta 9-THC according to the proposed pharmacophore overlay, their potencies are predicted by a quantitative model of cannabinoid structure--activity relationships based solely on classical and nonclassical cannabinoids with a reasonable degree of accuracy. The ability to incorporate the pharmacological potency of these anandamides into the cannabinoid pharmacophore model is also shown to support the relevance of the proposed pharmacophore model.

Pharmacological specificity of the discriminative stimulus effects of Δ9-tetrahydrocannabinol in rhesus monkeys

Δ 9-Tetrahydrocannabinol ( Δ 9-THC) discrimination has been used as an animal model of cannabis intoxication in humans. While numerous studies have examined the discriminative stimulus effects of cannabinoids in rats and pigeons, studies with monkeys have been rare. In the present study, rhesus monkeys, trained to discriminate Δ 9-THC from vehicle in a two-lever drug discrimination procedure, were tested with a variety of psychoactive drugs, including cannabinoids as well as drugs from other classes. Results showed that Δ 9-THC discrimination showed pharmacological specificity, in that none of the non-cannabinoid drugs fully substituted for Δ 9-THC. In contrast, the classical cannabinoids, Δ 9-THC and Δ 8-THC, and the novel cannabinoids, WIN 55212-2 and 1-butyl-2-methyl-3-(1-naphthoyl)indole, produced full dose-dependent substitution for Δ 9-THC in all monkeys. A heptyl indole derivative failed to substitute for Δ 9-THC, but it also did not displace [ 3H] CP 55940 from its binding site. These findings are consistent with those of previous cannabinoid discrimination studies with rats and suggest that results of Δ 9-THC discrimination studies in rhesus monkeys may be predictive of the subjective effects of cannabinoid drugs in humans.

Aminoalkylindoles: structure-activity relationships of novel cannabinoid mimetics.

Aminoalkylindoles (AAIs) are a novel series of cannabinoid receptor ligands. In this report we disclose the structural features of AAIs which are important for binding to this receptor as measured by inhibition of binding of [3H]Win 55212-2 (5). Functional activity in the mouse vas deferens is also noted and used to distinguish agonists from potential antagonists. The key structural features for potent cannabinoid activity in this series are a bicyclic (naphthyl) substituent at the 3-position, a small (H) substituent at the 2-position, and an aminoethyl (morpholinoethyl) substituent at the 1-position. A 6-bromo analog, Win 54461 (31), has been identified as a potential cannabinoid receptor antagonist. Modeling experiments were done to develop a pharmacophore and also to compare AAI structures with those of classical cannabinoids. The fact that the cannabinoid AAIs arose out of work on a series of cyclooxygenase inhibitors make sense now that an endogenous cannabinoid ligand has been identified which is a derivative of arachidonic acid. Because of their unique structures and physical properties, AAIs provide useful tools to study the structure and function of the cannabinoid receptor(s).

Common cannabimimetic pharmacophoric requirements between aminoalkyl indoles and classical cannabinoids

Aminoalkylindoles (AAIs) are structurally dissimilar from the classical cannabinoids (CCs), however, both AAIs and CCs appear to bind at the same site on the cannabinoid receptor. To obtain better insights on the structural correlation between AAIs and CCs, we have studied the conformational properties of the potent cannabimimetic AAI WIN 55212-2 and its inactive analogs using high resolution 2D NMR spectroscopy in combination with computer-assisted molecular modeling. The pharmacophoric similarities between the AAIs and the CCs were then investigated using superimposition techniques. The absolute stereochemistries of the biologically active enantiomer (-)HHC were used as superimposition points and considered as internal controls in order to test the molecular principles guiding this experiment. Our results show that the model is congruent with a superimposition in which the naphthoyl, morpholino and 3-keto groups in the AAI, respectively correspond to the side chain, cyclohexanol OH and phenolic OH of HHC. A good fit is obtained when the two biologically active antipodes are superimposed. Conversely, the fit is poor if the inactive AAI enantiomer is superimposed on the active HHC enantiomer. It can also be seen that in such an orientation a certain deviation of the C-ring from the plane of the phenol ring of the tricyclic HHC component and of the morpholinyl portion from the plane of the indole ring of WIN 55212-2 is essential for cannabimimetic activity. The inactive enantiomer WIN 55212-3 has its respective components aligned in the opposite quadrant. By comparing the stereoelectronic features of representative AAIs and CCs, we have developed a model which may help to uncover the pharmacophoric requirements of the AAIs and serve as a basis for future SAR and drug design.

Characterization of a region of steric interference at the cannabinoid receptor using the active analog approach.

In this paper, it is hypothesized that the distinction between certain active and inactive cannabinoids is that the inactive analogs possess extra volume associated with their carbocyclic rings that may be responsible for an unfavorable interaction at the cannabinoid receptor. Using the active analog approach, a model is developed of a region of steric interference at this receptor using the active cannabinoids (-)-trans-delta 9-tetrahydrocannabinol, (-)-trans-delta 8-tetrahydrocannabinol, (-)-11-hydroxy-beta-hexahydrocannabinol, and a (-)-trans-11-hydroxy-delta 8-tetrahydrocannabinol dimethylheptyl derivative and the inactive cannabinoids (9S,6aR)-trans-delta 10,10a-tetrahydrocannabinol and a (+)-trans-11-hydroxy-delta 8-tetrahydrocannabinol dimethylheptyl derivative. Each of these molecules satisfy the cannabinoid pharmacophoric requirements, i.e., a phenolic oxygen at C1 and a side chain of acceptable length at C3. Accessible conformers of each molecule were identified by using the method of molecular mechanics as encoded in the MMP2(85) program. The MAP facility within the Chem-X molecular modeling program was then used to calculate the region of steric interference (termed the receptor essential volume, REV) from these accessible conformers. The calculations revealed an REV region located near the top of the carbocyclic ring in the bottom face of the molecule. In order to explore the use of this REV to account for the activities of other cannabinoids, the minimally active classical cannabinoid (-)-11-hydroxy-alpha-hexahydrocannabinol, an active benzofuran cannabinoid, and the active nonclassical cannabinoid CP-47,497 were then studied. In each case, the activity or minimal activity of each compound can be explained on the basis of the ability of one or more accessible conformer of each molecule to clear the REV calculated here. The results of this study provide an explanation at the molecular level for observed activity differences between cannabinoids that exhibit shape differences associated with their carbocyclic rings.

Behavioral, biochemical, and molecular modeling evaluations of cannabinoid analogs

Numerous cannabinoids have been synthesized that are extremely potent in all of the behavioral assays conducted in our laboratory. An important feature in increasing potency has been the substitution of a dimethylheptyl (DMH) side chain for the pentyl side chain. Our previous studies have shown that (−)-11-OH- Δ 8-THC-dimethylheptyl was 80–1150 times more potent than Δ 9-THC. Stereospecificity was demonstrated by its (+)- enantiomer which was more than 1400–7500 times less potent. A related series of DMH cannabinoid analogs has recently been synthesized and preliminary evaluations reported here. (−)-11-OH- Δ 9-THC-DMH was found to be equipotent with (−)-11-OH- Δ 8-THC-DMH. The aldehyde (−)-11-oxo- Δ 9-THC-DMH was 15–50 times more potent than Δ 9-THC. Surprisingly, (−)-11-carboxy- Δ 9-THC-DMH was also active, being slightly more potent than Δ 9-THC. In the bicyclic cannabinoid series, the length and bulk of the side chain were found to be equally important. Aminoalkylindoles, which are structurally dissimilar from classical cannabinoids, have been found to exhibit a pharmacological profile similar to Δ 9-THC. Though not extremely potent in vivo, they appear to represent an entirely new approach to studying the actions of the cannabinoids. The structural diversity and wide-ranging potencies of the analogs described herein provide the opportunity to develop a pharmacophore for the cannabinoids using molecular modeling techniques.

A molecular reactivity template for cannabinoid activity

Methods of theoretical chemistry are used to characterize the molecular structure and some reactivity properties of the major psychopharmacologically active component of cannabis, (—)-trans-Δ 9-tetrahydrocannabinol (Δ 9-THC). This characterization is part of an exploration of the molecular parameters that could serve to unify considerations of structure-activity relationships in disparate classes of cannabinoids. Our hypothesis is that there are two components of the Δ 9-THC structure that confer upon the molecule reactivity characteristics crucial to activity: the lone pairs of electrons of the phenol oxygen, and the orientation of the carbocyclic ring. The conformation of Δ 9-THC is calculated using molecular mechanics. Since the position of the phenolic OH is central to the working hypothesis, the rotational energy behavior of the phenolic OH is studied. Results from the calculations identify two minimum energy conformations of the OH in Δ 9-THC. Results from ab-initio calculations of the OH rotation on a model fragment of Δ 9-THC agree well with the molecular mechanics results. The molecular electrostatic potential (MEP) of Δ 9-THC is calculated for the energetically optimal conformations. The results indicate that the two faces of the Δ 9-THC molecule are distinguishable. The MEP of the bottom face of Δ 9-THC (in each of the two minimum energy conformations of the phenolic OH), along with the conformational results for the orientation of the carbocyclic ring relative to the phenol group form a reactivity template to be used in comparisons with properties of active and inactive cannabinoids.