Effects of Δ9-Tetrahydrocannabinol and the Aminoalkylindole K2/Spice Constituent JWH-073 on Cardiac Tissue and Mesenteric Vascular Reactivity.

Background: Although use of Cannabis sativa is not associated with serious adverse effects, recreational use of aminoalkylindole (AAI) cannabinoid receptor agonists found in K2/Spice herbal blends has been reported to cause adverse cardiovascular events, including angina, arrhythmia, changes in blood pressure, ischemic stroke, and myocardial infarction. Δ9-Tetrahydrocannabinol (Δ9-THC) is the primary CB1 agonist found in cannabis and JWH-073 is one of the AAI CB1 agonists found in K2/Spice brands sold to the public. Methods: This study used in vitro, in vivo, and ex vivo approaches to investigate potential differences on cardiac tissue and vascular effects betweenJWH-073 and Δ9-THC. Male C57BL/6 mice were treated with JWH-073 or Δ9-THC and cardiac injury was assessed by histology. Effects of JWH-073 and Δ9-THC on H9C2 cell viability and ex vivo mesenteric vascular reactivity were also determined. Results: JWH-073 or Δ9-THC induced typical cannabinoid effects of antinociception and hypothermia but did not promote death of cardiac myocytes. No differences in cell viability were observed in cultured H9C2 cardiac myocytes after 24 h of treatment. In isolated mesenteric arteries from drug-naive animals, JWH-073 produced significantly greater maximal relaxation (96%±2% vs. 73%±5%, p<0.05) and significantly greater inhibition of phenylephrine-mediated maximal contraction (Control 174%±11%KMAX) compared with Δ9-THC (50%±17% vs. 119%±16%KMAX, p<0.05). Discussion: These findings suggest that neither cannabinoid at the concentrations/dose studied caused cardiac cell death, but JWH-073 has the potential for greater vascular adverse events than Δ9-THC through an increased vasodilatory effect.

Comparison of the Neurotoxic and Seizure-Inducing Effects of Synthetic and Endogenous Cannabinoids with Δ9-Tetrahydrocannabinol.

Introduction: Synthetic cannabinoids (SCs) are commonly found in preparations used as recreational drugs. Although severe adverse health effects are not generally associated with cannabis use, a rising number of studies document seizures and even death after SC use. In this study, a mouse model is used to investigate the hypothesis that SCs are more toxic than Δ9-tetrahydrocannabinol (THC), the principal psychoactive constituent of cannabis. Materials and Methods: Beginning with the SCs, JWH-073 and AM-2201, dose-response curves were generated to find the dose of each drug that was similarly efficacious to 50 mg/kg THC. Mice were given daily intraperitoneal (IP) injections of vehicle, 50 mg/kg THC, 30 mg/kg JWH-073, or 1 mg/kg AM-2201 until tolerance to the antinociceptive and hypothermic effects was complete, and then were assessed for spontaneous and antagonist-precipitated withdrawal and potential organ damage. No differences in tolerance were noted, but AM-2201 showed more rearing in the spontaneous and antagonist-precipitated withdrawal phases than either vehicle or the other two drug treatments. Histopathological examination of these mice revealed no drug-induced lesions. In a subsequent set of experiments, various doses of THC, methanandamide (mAEA), and of a variety of SCs (HU-210, CP55940, JWH-073, AM-2201, and PB-22) were given IP, and convulsions and change in body temperature were quantified. Discussion: The treatments yielded varying numbers of convulsions and a range of changes in body temperature. JWH-073 and AM-2201 produced significantly more convulsions than THC, HU-210, mAEA, or cannabidiol (CBD) (the latter two producing none). HU-210, CP55940, JWH-073, and mAEA produced greater hypothermia than THC or CBD. Convulsions and hypothermia induced by several agonists were prevented by pretreatment with a CB1 antagonist, but not a CB2 antagonist. Conclusions: In agreement with human studies and case reports, this study found that SCs generally produced more seizures than THC. Of particular significance was the finding that mAEA produced far greater hypothermia than THC (similar to most SCs), but unlike the SCs and THC, produced no seizures.

Interdisciplinary pharmaceutical sciences activity within a pharmacy practice skills course.

BACKGROUND AND PURPOSE: To help students develop a more integrated mindset, an integrated curricular model can facilitate the building of connections between the foundational and clinical sciences by presenting multi-disciplinary material within a cohesive framework in a single course. The main objective of this research was to assess the impact of interdisciplinary teaching on student performance in a skills course. EDUCATIONAL ACTIVITY AND SETTING: A case study was presented and questions were embedded through an audience response system. Each of three groups of students (approximately 32 students per group) were divided into teams, and the scores were shown periodically to produce an atmosphere of friendly competition. The entire exercise lasted approximately 50 min. FINDINGS: Students found the pharmaceutical science and pharmacy practice faculty collaboration helpful in regards to reviewing for integrated exams. Student pharmacists were asked to provide one positive aspect of the course and one area for improvement. Twenty-two of 96 responders indicated that the integrated class session was the highlight of the course. Student pharmacists noted that they were able to recognize the integration between the basic and applied sciences. Students clearly favored this learning style over the didactic approach, as evidenced by the feedback. In the future, we plan to implement longer integrated cases more frequently to train future pharmacists as critical and integrative thinkers. SUMMARY: This integrated case study appeared to be effective in helping student pharmacists apply knowledge of various basic science disciplines to the applied sciences.

Investigation of non-CB1, non-CB2 WIN55212-2-sensitive G-protein-coupled receptors in the brains of mammals, birds, and amphibians.

PURPOSE: Previous studies have found non-CB1 non-CB2 G-protein-coupled receptors in rodents that are activated by the aminoalkylindole cannabinoid agonist WIN55212-2. This work obtained evidence for the presence or absence of similar receptors in the brains of other mammals, birds and amphibians. MATERIALS AND METHODS: Antagonism of the stimulation of [35S]GTPγS binding by WIN55212-2 and CP55940 was assessed in multiple CNS regions of rat and canine, and in whole brain membranes from shrew, pigeon, frog and newt. A bioinformatics approach searched for orthologs of GRP3, GPR6, and GPR12 (closely related to cannabinoid receptors) in the genomes of these or related species. Orthologs were examined for amino acid motifs known to impart functionality to receptors. RESULTS: In mammals and pigeon, but not amphibians, a significant fraction of the stimulation of [35S]GTPγS binding by WIN55212-2 was not blocked by the CB1 antagonist SR141716A. BLAST searches found that GPR3 was restricted to mammals. GPR12 orthologs existed in all species, and they shared identical amino acid motifs. GPR6 orthologs existed all species, but with significant departures in the identity of some critical amino acids in bird, more so in amphibian. CONCLUSIONS: The portion of WIN55212-2-stimulated [35S]GTPγS binding that was antagonized by SR141716A was consistent with stimulation via CB1 receptors, indicating that antagonist-insensitive activity was via a different G-protein coupled receptor. Pharmacological evidence of this receptor was found in the brains of mammals and pigeon, but not frog or newt. Bioinfomatics results implicate GPR6 as a possible candidate for the additional WIN55212-2-sensitive receptor.

The effects of beta-arrestin1 deletion on acute cannabinoid activity, brain cannabinoid receptors and tolerance to cannabinoids in mice.

CONTEXT: Previous studies have indicated a role for beta-arrestin2 in the regulation of brain cannabinoid effects and cannabinoid CB1 receptors, but whether beta-arrestin1 has a role has not been investigated. OBJECTIVE: To determine the role of beta-arrestin1 in cannabinoid activity. MATERIALS AND METHODS: Beta-arrestin1 -/- mice and their wild-type (+/+) counterparts were assayed for antinociceptive and temperature-decreasing effects of two ligands, Δ(9)-tetrahydrocannabinol (THC) and CP55940, after both single and repeated administration. In vitro assays examined the effects of deletion on CB1 receptor density, agonist-binding and G-protein activation. RESULTS: Deletion of beta-arrestin1 diminished the effects of CP55940 in both antinociception (latency to tail withdrawal) and temperature-depression assays in mice. However, deleting beta-arrestin1 had no effect on the actions of THC in either assay. Antagonist radioligand ([(3)H]SR141716A) saturation binding indicated no difference between beta-arrestin1 +/+ and -/- mice in the density or affinity for cannabinoid CB1 receptors in brain membranes. CP55940 agonist binding in brain membranes from beta-arrestin1 +/+ mice exhibited high- and intermediate-affinity sites, but beta-arrestin1 -/- membranes exhibited an additional site with low affinity. CP55940 produced greater stimulation of [(35)S]GTPγS binding to membranes from whole brain of beta-arrestin1 -/- than +/+ mice. The rates of the development of tolerance to chronic THC or CP55940 administration did not appear to be affected by genotype. DISCUSSION: Beta-arrestin1 appeared to mediate the actions of CP55940, but did not affect the activity of THC. CONCLUSION: Beta-arrestin1 regulates cannabinoid CB1 receptor sensitivity in an agonist-selective manner, but may not be the primary mediator of tolerance to cannabinoid agonists.

Basic neuroanatomy and neuropharmacology of cannabinoids.

Humans have used Cannabis sativa (marijuana) for at least 12,000 years, but researchers have only recently described an endogenous cannabinoid system. The endocannabinoid system modulates an array of physiological and psychological functions. Endocannabinoids are widely distributed throughout the body, including the central nervous system (CNS). This article gives a basic overview of endocannabinoid neuroanatomy and function. Several endocannabinoids have been discovered to date, and their roles are being elucidated. Two G-protein coupled cannabinoid receptors, CB1R and CB2R, have been identified, although other candidate receptors exist, including ion channel and nuclear receptors that might be components of the endocannabinoid system. It appears that cannabinoids are dysregulated in a number of psychiatric disorders and might be involved in their pathogenesis. There is now evidence that manipulation of the endocannabinoid system could be a therapeutic target for a variety of conditions.