Effects of chronic administration of delta-9-tetrahydrocannabinol and cannabidiol on rat testicular esterase isozymes

The effects of Δ⁹-tetrahydrocannabinol and cannabidiol on the esterase isozymes of rat testicular tissue were examined using acrylamide gel electrophoretic techniques followed by staining for non-specific esterase activity. Both Δ⁹-tetrahydrocannabinol and cannabidiol caused a specific depression of an esterase isozyme located in the interstitial tissue of the testis.     

Hydroxylation and glycosylation of Δ9-tetrahydrocannabinol by Catharanthus roseus cell suspension culture

Δ⁹-tetrahydrocannabinol is the active constituent in Cannabis sativa, with reported analgesic, anti-emetic, anti-oxidative, neuroprotective, and anti-inflammatory activities. Δ⁹-THC has been used to treat a number of disease states including pain, anxiety, asthma, glaucoma, and hypertension. Poor water solubility of Δ⁹-THC greatly reduces its clinical effectiveness. Consequently, there is a need to modify the compound to increase its polarity and pharmaceutical efficacy. The aim of this study was to test the capability of Catharanthus roseus suspension cultured cells to convert Δ⁹-THC into more polar derivatives. The transformed metabolites were analyzed and isolated by HPLC. Structures of some new derivatives were proposed on the basis of molecular ion peaks and fragmentation patterns obtained from LC-MS and UV spectra obtained by HPLC, respectively. Δ⁹-THC was rapidly absorbed by Catharanthus roseus cultured cells and upon biotransformation new glycosylated and hydroxylated derivatives were isolated by preparative HPLC. In addition, cannabinol was detected as degradation product, including its glycosylated derivative. Based on these results, it is concluded that Catharanthus cultured cells have great potential to transform Δ⁹-THC into more polar derivatives and can be used for the large scale production of new cannabinoids, which can be a source of new compounds with interesting pharmacological profiles.     

Hypothermic action of delta 9-tetrahydrocannabinol, 11-hydroxy-delta 9-tetrahydrocannabinol and 11-hydroxy-delta 8-tetrahydrocannabinol in mice

The hypothermic activity of Δ⁹-tetrahydrocannabinol (Δ⁹-THC), a metabolite, 11-hydroxy-Δ⁹-tetrahydrocannabinol (11-OH-Δ⁹-THC) and 11-hydroxy-Δ⁸-tetrahydrocannabinol (11-OH-Δ⁸-THC) has been determined in male mice maintained at an ambient temperature of 20 ± 1°C. The mean body temperature of mice that received 2, 4, 16 or 32 mg/kg, i. v., of a tetrahydrocannabinol was significantly lower than that of vehicle treated mice (p <0.05) within 2 minutes after drug administration. Dose-response relationships show the intrinsic activity of Δ⁹-THC to be significantly greater than that of 11-OH-Δ⁹-THC or 11-OH-Δ⁸-THC in this system (p <0.05). The data indicate that the hypothermic activity of Δ⁹-THC cannot be explained entirely by metabolism to 11-OH-Δ⁹-THC.     

Influence of anticonvulsant cannabinoids on posttetanic potentiation at isolated bullfrog ganglia

The effects of anticonvulsant cannabinoids on posttetanic potentiation (PTP) at bullfrog paravertebral ganglia $\text{in vitro}$ were investigated electrophysiologically. Two Δ⁹-tetrahydrocannabinol metabolites, 11-hydroxy-Δ⁹-tetrahydrocannabinol and 8α, 11-dihydroxy-Δ⁹-tetrahydrocannabinol, as well as cannabidiol, markedly depressed PTP. In contrast, Δ⁹-tetrahydrocannabinol had no such effect. Thus, the findings of the present study clearly demonstrate that pharmacological properties of these hydroxylated metabolites of Δ⁹-tetrahydrocannabinol are not identical to those of their parent compound.     

Greenhouse propagation of Cannabis Sativa L. by vegetative cuttings

Previous work revealed significant variations in cannabinoid profiles ofCannabis sativa L. derived from a single seed source (P.1. 378939) and subjected to the same growth environment. Studies were conducted to evaluate the efficacy of propagation ofC. sativa by vegetative cuttings in order to increase uniformity of cannabinoid concentrations within a given plant population.C. sativa was successfully propagated by vegetative cuttings. However, there were both morphological and biochemical differences between seed-derived plants and their vegetative propagules. Delta-⁹-tetrahydrocannabinol concentrations were 4.1 times higher in vegetative propagules than in seed propagules. Vegetative cuttings also generally developed more profuse lateral branch growth; hence, foliage increased relative to their parent plants. Cannabinoid levels within the population of vegetative cuttings remained highly variable.     

The effects of Δ9-tetrahydrocannabinol Δ9-THC) on the regional concentrations of spermidine in the rat brain

The effects of intravenous administration of Δ⁹-tetrahydrocannabinol (Δ⁹-THC, 2 mg/kg, i.v.) on the regional brain spermidine concentrations of the rat were examined. Thirty minutes after vehicle treatment, the spermidine concentrations were: for the medulla oblongata/pons, 68.2 ± 7.7 μg/g; the hypothalamus, 67.7 ± 2.6 μg/g; the midbrain, 59.1 ± 4.4 μg/g; the cerebellum, 47.3 ± 5.9 μg/g and for the cortex, 13.8 ± 0.8 μg/g. Thirty minutes after Δ⁹-THC, these concentrations were reduced in the midbrain (47.0 ± 8.0% of control, P < 0.0001) and cortex (69.4 ± 7.4% of control, P < 0.009). The spermidine concentrations were not significantly altered in the medulla oblongata/pons (86.5 ± 13.3%, P > 0.36), hypothalamus (107.2 ± 11.8, P > 0.36) or cerebellum (89.0 ± 14.4%, P < 0.48). These results suggest that spermidine within the midbrain and cortex may be involved in the expression of some of the actions of Δ⁹-THC.     

Inhibition of sea urchin fertilization by fatty acid ethanolamides and cannabinoids

The influence of saturated and unsaturated fatty acid ethanolamides as well as Δ⁹-tetrahydrocannabinol (Δ⁹-THC), WIN 55,212-2 and cannabinoid CB₁ receptor antagonist SR 141716 on sea urchin fertilization was studied. The ethanolamides of arachidonic, oleic and linoleic acids but not saturated fatty acid (C₁₄–C₂₀) derivatives inhibited fertilization when pre-incubated with sperm cells. Δ⁹-THC and WIN 55,212-2 also inhibited fertilization, Δ⁹-THC being ten times as potent as WIN 55,212-2. Selective cannabinoid CB₁ receptor antagonist SR 141716 also blocked fertilization and did not antagonize the action of Δ⁹-THC. The obtained results indicate that different unsaturated fatty acid ethanolamides may control sea urchin fertilization, and that sea urchin sperm cell cannabinoid receptor may differ from the known cannabinoid receptor subtypes.     

Acquisition of tolerance to Δ-9-THC as measured by the response of a cellular function

The development of tolerance to delta-9-tetrahydrocannabinol (Δ-9-THC) was investigated by measuring respiration in brain tissue after acute or chronic administration. Mice were given either single or seven daily repeated intraperitoneal injections of 50 mg/Kg of delta-9-tetrahydrocannabinol (Δ-9-THC) or control vehicle. The final injection for all drug treated animals included radiolabeled ³H-Δ-9-THC. The mice were sacrificed at 1 hour, 2 hours, 4 hours, 24 hours, and 7 days after the final injection. Δ-9-THC depressed respiration, but after repeated injections was significantly less effective in this regard, indicating acquisition of tolerance to Δ-9-THC. Because the concentration of radiolabeled cannabinoids in brain tissue from each group is not appreciably different, a cellular as opposed to distributional mode of tolerance is suggested.     

The Genetic Structure of Marijuana and Hemp

Despite its cultivation as a source of food, fibre and medicine, and its global status as the most used illicit drug, the genus Cannabis has an inconclusive taxonomic organization and evolutionary history. Drug types of Cannabis (marijuana), which contain high amounts of the psychoactive cannabinoid Δ ⁹-tetrahydrocannabinol (THC), are used for medical purposes and as a recreational drug. Hemp types are grown for the production of seed and fibre, and contain low amounts of THC. Two species or gene pools (C. sativa and C. indica) are widely used in describing the pedigree or appearance of cultivated Cannabis plants. Using 14,031 single-nucleotide polymorphisms (SNPs) genotyped in 81 marijuana and 43 hemp samples, we show that marijuana and hemp are significantly differentiated at a genome-wide level, demonstrating that the distinction between these populations is not limited to genes underlying THC production. We find a moderate correlation between the genetic structure of marijuana strains and their reported C. sativa and C. indica ancestry and show that marijuana strain names often do not reflect a meaningful genetic identity. We also provide evidence that hemp is genetically more similar to C. indica type marijuana than to C. sativa strains.     

Delta9-tetrahydrocannabinol: subcortical spike bursts and motor manifestations in a Fischer rat treated orally for 109 days

A total of 12 Fischer rats was prepared surgically for chronic EEG recording from cortical and subcortical sites. Most rats, within 2 to 9 weeks after electrode implantation, developed polyspike activity in cortical and subcortical recordings that were without motor manifestations. Six of these rats, chronically treated po with Δ⁹-tetrahydrocannabinol (Δ⁹-THC) 10 mg/kg exhibited acute EEG changes with more frequent occurrence of EEG desynchronization and polyspike activity. On day 109 one of 6 rats displayed consulsive activity, with jerky movements of the head and paws, characteristics of Δ⁹-THC neurotoxicity. EEG alterations concomitant with motor signs included bursts of spikes of approximately 0.2 sec that occurred in subcortical, but not in cortical, recordings. It is concluded that in the Fischer rat acute and chronic treatment with Δ⁹-THC facilitated the occurrence of surgically-induced “polyspike” activity while chronic treatment caused occasional transient subcortical spike bursts with concomitant motor manifestations.     

Systemic Injections of Cannabidiol Enhance Acetylcholine Levels from Basal Forebrain in Rats

Cannabis sativa is a plant that contains more than 500 components, of which the most studied are Δ⁹-tetrahydrocannabinol (Δ⁹-THC) and cannabidiol (CBD). Several studies have indicated that CBD displays neurobiological effects, including wake promotion. Moreover, experimental evidence has shown that injections of CBD enhance wake-related compounds, such as monoamines (dopamine, serotonin, epinephrine, and norepinephrine). However, no clear evidence is available regarding the effects of CBD on additional wake-related neurochemicals such as acetylcholine (ACh). Here, we demonstrate that systemic injections of CBD (0, 5, 10 or 30 mg/kg, i.p.) at the beginning of the lights-on period, increase the extracellular levels of ACh collected from the basal forebrain and measured by microdialysis and HPLC means. Moreover, the time course effects on the contents of ACh were present 5 h post-injection of CBD. Altogether, these data demonstrate that CBD increases ACh levels in a brain region related to wake control. This study is the first to show the effects of ACh levels in CBD-treated rats and suggests that the basal forebrain might be a site of action of CBD for wakefulness modulation.     

Sapogenins of Yucca glauca seed pods

From the fruiting pods of Yucca glauca Nutt. devoid of seeds the following sapogenins have been isolated and identified: neo-tigogenin, hecogenin, gitogenin, manogenin, Δ⁹-manogenin and sarsasapogenin. A small amount of a material believed to be Δ²-desoxysarsasapogenin was also isolated and is apparently an artifact arising from the hydrolysis and extraction procedure. Manogenin and Δ⁹-manogenin have not been previously detected in Y. glauca, and Δ⁹-manogenin has not been reported in any Yucca species.     

Anticonvulsant properties of delta 9-tetrahydrocannabinol and other cannabinoids

Anticonvulsant doses of Δ⁹-tetrahydrocannabinol (Δ⁹-THC) markedly lower body temperature in mice at an ambient temperature of 22°C, but there is little such effect at 30°C. The anticonvulsant properties of Δ⁹-THC are as follows: The drug abolishes hind-limb extension in a maximal electroshock (MES) test, elevates both the MES (extensor) and 6-Hz-electroshock thresholds, exerts no effect on the 60-Hz-electroshock threshold, and enhances minimal seizures caused by pentylenetetrazol. All anticonvulsant properties studied, with the exception of the 60-Hz-electroshock threshold, were unaffected by the hypothermia resulting at 22°C. Additional experiments with Δ⁹-THC indicated that chronic treatment results in the development of tolerance, as determined by the MES test with rats. The four principal naturally occurring cannabinoids, Δ⁹-THC, Δ⁸-THC, cannabinol and cannabidiol, display anticonvulsant activity, as does the major, primary metabolite of Δ⁹-THC, 11-hydroxy-Δ⁹-THC. Of all agents investigated in mice, the synthetic cannabinoids, dimethylheptylpyran and its isomers, are the most potent anticonvulsants. The results of a study of the relative motor toxicity and anticonvulsant activity of the cannabinoids demonstrate that these properties are at least partially separable among the various agents.     

Effects of acute and chronic administration of Cannabis sative extract on the mouse-killing behavior of rats

The muricidal behavior of spontaneous killer rats was blocked after the first injections of an extract of $\text{Cannabis sativa}$ corresponding to 8.8 mg of Δ9-tetrahydrocannabinol. On the contrary, to rats which did not show this behavior spontaneously, the chronic administration of the extract induced mouse-killing behavior; this effect of marihuana was potentiated by starvation which, by itself, did not interfere with the behavior of the non-killer rats. These results tend to support the view that cannabis has a dual action. Suppression of muricidal behavior may occur due to the acute depressant effects of marihuana. Under chronic administration, however, tolerance develops to these effects unmasking hyperemotionality and/or irritability which may induce killing behavior in previously non-killer rats.     

Cannabinoid formation in Cannabis sativa grafted inter-racially,and with two Humulus species

Inter-racial grafts between high and low Δ¹-THC strains of Cannabis sativa, as well as cross-grafts with two Humulus (hop) species have been effected. C. sativa strains continue to produce essentially their own characteristic mixtures of cannabinoids, with undiminished vigour, whatever part of the graft system they form. There is no evidence of transport of intermediates or factors critical to cannabinoid formation across the grafts.     

Δ9-tetrahydrocannabinol and dimethylheptylpyran induced tachycardia in the conscious rat

Since cannabinoids lead to dose-related tachycardia in man but dose dependent bradycardia has been reported thus far in laboratory animals, there would seem to be a need for an experimental model in which the effect seen in man (tachycardia) could be reproduced and explored. In the conscious rat, the compounds Δ⁹-tetrahydrocannabinol (Δ⁹-THC) and dimethylheptylpyran (DMHP) injected i.p. led to dose-related increases in heart rate at 10–20 minutes after administration. In vehicle (ethanol) control rats there were small increases in heart rate. Propranolol given before Δ⁹-THC resulted in a parallel shift to the right of the dose-effect curve. Adrenalectomy led to a significant (p<0.01) decrease in tachycardia following Δ⁹-THC and DMHP while ganglionic block markedly decreased the heart rate increases after Δ⁹-THC (p<0.001). Systolic blood pressure at nearly all doses of Δ⁹-THC was minimally affected, although it tended to decrease with increasing dose. Tachycardia in the rat may be the result of a centrally mediated release of epinephrine from the adrenal gland.     

Purification of Δ9-Tetrahydrocannabinol,an Active Constituent of Marihuana,by Accelerated Microparticulate Gel Chromatography

A technique is described by which Δ⁹ -tetrahydrocannabinol, the principal psychotomimetic constituent of marihuana and hashish, can be isolated in a chromatographically pure state from synthetic or natural products by pressure-accelerated chromatography through columns of ultrafine silica gel packed by centrifugal force.     

Distinct pharmacology and metabolism of K2 synthetic cannabinoids compared to Δ-THC: Mechanism underlying greater toxicity?

K2 or Spice products are emerging drugs of abuse that contain synthetic cannabinoids (SCBs). Although assumed by many teens and first time drug users to be a “safe” and “legal” alternative to marijuana, many recent reports indicate that SCBs present in K2 produce toxicity not associated with the primary psychoactive component of marijuana, ?⁹-tetrahydrocannabinol (Δ⁹-THC). This mini-review will summarize recent evidence that use of K2 products poses greater health risks relative to marijuana, and suggest that distinct pharmacological properties and metabolism of SCBs relative to Δ⁹-THC may contribute to the observed toxicity. Studies reviewed will indicate that in contrast to partial agonist properties of Δ⁹-THC typically observed in vitro, SCBs in K2 products act as full cannabinoid receptor type 1 (CB1R) and type 2 (CB2R) agonists in both cellular assays and animal studies. Furthermore, unlike Δ⁹-THC metabolism, several SCB metabolites retain high affinity for, and exhibit a range of intrinsic activities at, CB1 and CB2Rs. Finally, several reports indicate that although quasi-legal SCBs initially evaded detection and legal consequences, these presumed “advantages” have been limited by new legislation and development of product and human testing capabilities. Collectively, evidence reported in this mini-review suggests that K2 products are neither safe nor legal alternatives to marijuana. Instead, enhanced toxicity of K2 products relative to marijuana, perhaps resulting from the combined actions of a complex mixture of different SCBs present and their active metabolites that retain high affinity for CB1 and CB2Rs, highlights the inherent danger that may accompany use of these substances.     

Allylbenzene analogs of Δ9-tetrahydrocannabinol as tumor growth inhibitors

A series of substituted allylbenzene derivatives was tested $\text{in vitro}$ for ability to inhibit cell growth in the KB cell line. None of the compounds was as active as Δ⁹-tetrahydrocannabinol. It is suggested that inhibition of cell growth by cannabinoids requires a polycyclic system which may act by an intercalaction mechanism.     

Endocrine effects of chronic intraventricular administration of delta9-tetrahydrocannabinol to prepuberal and adult male rats.

The daily intraventricular administration of Δ⁹-tetrahydrocannabinol (Δ⁹-THC) in microgram amounts for a week to prepuberal and adult rats had definite endocrine effects. Prostate weights were reduced and plasma and pituitary levels of growth hormone (GH) were increased in prepuberal rats. Pituitary levels of prolactin (PRL) were increased both in prepuberal and in adult animals while pituitary and adrenal weights and plasma corticosterone (B) levels were increased in adult rats. On the other hand, brain weights were significantly reduced by Δ⁹-THC in prepuberal and significantly increased in adult animals. No changes in brain levels of noradrenaline (NA), dopamine (DA) or serotonin (5-HT) were found in treated animals. These results indicate that Δ⁹-THC may modify some endocrine functions when injected directly into the brain in microgram amounts. They show on the other hand that young and adult animals may respond differently to the chronic administration of the psychoactive drug, although the difference may be due to a biphasic effect of different doses.