Article

Pharmacological and therapeutic targets for Δ 9 tetrahydrocannabinol and cannabidiol

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Abstract

Summary Cannabis is the unique source of a set of at least 66 compounds known collectively as cannabinoids. Of these, most is known about the pharmacology of 9-tetrahydrocannabinol (9-THC), the main psychoactive constituent of cannabis, and about cannabidiol (CBD), which lacks psychoactivity. Accordingly, this paper focuses on the pharmacological and therapeutic targets of these two cannabinoids. Many of the effects of 9-THC are mediated by cannabinoid receptors of which at least two types, CB1 and CB2, are present in mammalian tissues. Endogenous agonists for cannabinoid receptors have also been discovered. CB1 receptors are present at the terminals of central and peripheral neurones, where they modulate transmitter release. They also exist in some non-neuronal cells. CB2 receptors are expressed mainly by immune cells, one of their roles being to alter cytokine release. 9-THC also appears to have non-CB1, non-CB2 pharmacological targets. It is already licensed for clinical use in the U.S.A. as an anti-emetic and appetite stimulant and both 9-THC and 9-THC-rich cannabis extracts show therapeutic potential as neuroprotective and anticancer agents and for the management of glaucoma, pain and various kinds of motor dysfunction associated, for example, with multiple sclerosis and spinal cord injury. CBD has much less affinity for CB1 and CB2 receptors than 9-THC and its pharmacological actions have been less well characterized. Potential clinical applications of CBD and CBD-rich cannabis extracts include the production of anti-inflammatory and neuroprotective effects, the management of epilepsy, anxiety disorders, glaucoma and nausea, and the modulation of some effects of 9-THC.

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... Glyceollins (240)(241)(242) markedly suppress the LPSdependent expression of COX-2 in LPS-treated RAW264.7 cells [75,76]. Prenylated phloroglucinols mallotojaponin (359), mallotochromanol (360), butyrylmallotolerin (361), isobutyrylmallotochromanol (362), and isomallotochromanol (363) exhibit an inhibitory effect against the production of PGE 2 in LPSstimulated RAW264.7 cell line. ...
... Echinoisoflavanon (228), isolated from Echinosophora koreensis Nakai, inhibits NO production as well as iNOS enzyme activity in LPS-induced RAW264.7 cells [168]. Glabridin (223) shows dose-dependent moderate inhibition in NO levels in the same cellular system [174], similar to glyceollins (240)(241)(242), which suppressed the LPS-dependent secretion of NO in LPSactivated murine RAW264.7 cells through the inhibition of iNOS expression [75,76]. ...
... It is well known that cannabinoids interact with cannabinoid receptors, in which mainly CB 2 subtype has the importance for the correct function of immune system and control of inflammation [5,242]. Several clinical studies showed the effect of cannabinoids (Cannabis or extracts from Cannabis) on the Crohn´s disease or ulcerative collitis [242][243][244][245]. ...
Article
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... Synthetic cannabinoids (SCs) are psychoactive compounds pharmacologically related to the natural cannabinoids found in Cannabis sativa (i.e., Δ-9-tetrahydrocannabinol, Δ-8-tetrahydrocannabinol cannabinol, and cannabidiol [1,2]). The family of SCs is a chemically heterogenous class of designer psychoactive substances and comprises compounds with remarkable differences in their molecular structures and in their biophysico-chemical properties. ...
... Extracted chromatogram of APP-CHMINACA and the principal metabolites identified after 1 h by the incubation with MLM a and HLM b. APP-CHMINACA distal amide hydrolysis (M1), distal amide hydrolysis and mono-oxidation (CHM) (M4.[1][2][3][4][5] ...
Article
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The metabolic pathways of APP-CHMINACA were characterized to select the markers of intake for implementation into analytical assays used by the clinical and forensic communities. We have combined the evidences obtained by both in vitro experiments and administration studies on mice. APP-CHMINACA was incubated with either human or mouse liver microsomes. Urine and blood samples were collected at different time points from mice after injection of a 3 mg/kg dose of the test compound. Samples were analyzed using liquid chromatography-tandem mass spectrometry. The in vitro studies allowed to isolate eight different metabolic reactions, formed by two metabolic routes, with no differences between human and mouse liver microsomes. The main biotransformation route involved the hydrolysis of the distal amide group and the subsequent hydroxylation on the cyclohexyl-methyl ring. The second route involved multiple hydroxylation of the parent compound, followed by reduction to generate minor metabolites. In blood samples, the most abundant substances identified were APP-CHMINACA unchanged and the metabolites formed by the hydrolysis of the distal amide together with its hydroxylated products. In urine samples, four metabolites formed following the hydroxylation of the distal amide hydrolysis metabolite were detected as the most abundant and long-term metabolites. The outcomes of our study showed that the most suitable markers to detect the intake of APP-CHMINACA in blood and urine samples in the framework of toxicological, clinical and forensic investigations were the metabolite formed by the hydrolysis of the distal amide and its hydroxylated products.
... Entre los más de 66 cannabinoides conocidos (Pertwee, 2004;Lachenmeier y Walch, 2005) producidos por la planta encontramos: tetrahidrocannabinol (THC), cannabidiol (CBD), cannabinol (CBN), cannabichromene (CBC), cannabigerol (CBG) y al menos 60 otros cannabinoides (Turner et al., 1980). Canabidiol (CBD) es conocido como un ingrediente anti-psicoactivo ya que modera el efecto que tiene el THC (Roulac, 1997;Pertwee, 2004). ...
... Entre los más de 66 cannabinoides conocidos (Pertwee, 2004;Lachenmeier y Walch, 2005) producidos por la planta encontramos: tetrahidrocannabinol (THC), cannabidiol (CBD), cannabinol (CBN), cannabichromene (CBC), cannabigerol (CBG) y al menos 60 otros cannabinoides (Turner et al., 1980). Canabidiol (CBD) es conocido como un ingrediente anti-psicoactivo ya que modera el efecto que tiene el THC (Roulac, 1997;Pertwee, 2004). De la relación entre las concentraciones de THC, CBN y CBD, Fetterman et al. (1971) crearon una fórmula que determina el fenotipo de la planta. ...
... Despite CBD's low affinity for both CB1 and CB2 receptors [91], it interacts with CB1 receptors as a negative allosteric modulator [92]. Recent studies [93] have indicated that CBD functions as an inverse agonist of the CB2 receptor, potentially elucidating its anti-inflammatory properties given the expression of CB2 receptors in immune cells, with inhibition likely leading to reduced inflammation [94]. Unlike THC, CBD lacks psychoactive properties, rendering it valuable for the treatment of various disorders. ...
Article
Full-text available
The endogenous cannabinoid system (ECS) plays a critical role in the regulation of various physiological functions, including sleep, mood, and neuroinflammation. Phytocannabinoids such as Δ9-tetrahydrocannabinol (THC), cannabidiol (CBD), cannabinomimimetics, and some N-acylethanolamides, particularly palmitoyethanolamide, have emerged as potential therapeutic agents for the management of sleep disorders. THC, the psychoactive component of cannabis, may initially promote sleep, but, in the long term, alters sleep architecture, while CBD shows promise in improving sleep quality without psychoactive effects. Clinical studies suggest that CBD modulates endocannabinoid signaling through several receptor sites, offering a multifaceted approach to sleep regulation. Similarly, palmitoylethanolamide (PEA), in addition to interacting with the endocannabinoid system, acts as an agonist on peroxisome proliferator-activated receptors (PPARs). The favorable safety profile of CBD and PEA and the potential for long-term use make them an attractive alternative to conventional pharmacotherapy. The integration of the latter two compounds into comprehensive treatment strategies, together with cognitive–behavioral therapy for insomnia (CBT-I), represents a holistic approach to address the multifactorial nature of sleep disorders. Further research is needed to establish the optimal dosage, safety, and efficacy in different patient populations, but the therapeutic potential of CBD and PEA offers hope for improved sleep quality and general well-being.
... Cannabinoids are a group of more than 66 biologically active terpenophenolic compounds, derivatives of 2-substituted 5-amylresorcin, found in the cannabis plant (Cannabis sativa) [3] and its preparations, such as marijuana and hashish [4], as well as their synthetic analogues capable of binding to cannabinoid receptors [5]. ...
Article
Full-text available
Synthetic cannabinoids are a group of psychoactive compounds that mimic the effects of Δ9-tetrahydrocannabinol, the main psychoactive component of marijuana. Today, the most important task in forensic science is to establish the chemical structure of new psychoactive compounds that appear in illicit trafficking promptly in order to respond quickly and stop their distribution. Less important is the development of methodological support for expert activity, including analysis methods and reference data on the analytical characteristics of compounds. The aim. To develop stages of the forensic analysis of objects containing synthetic cannabinoids and propose methods for determining 5 new synthetic cannabinoids for forensic pharmaceutical purposes. Materials and methods. The study was conducted as part of the identification of cannabinoids for forensic purposes at the National Scientific Centre "Bokarius Institute of Forensic Examination". As part of the study, 5 new synthetic cannabinoids were identified for forensic analysis using the following methods: infrared spectroscopy, thin-layer chromatography, and gas chromatography with a mass detector. The algorithm for the forensic analysis of cannabinoid derivatives was developed based on the requirements of Ukraine's current legislation. Results. Spectral and chromatographic methods of determination of 5 new synthetic cannabinoids for forensic purposes were proposed, and during research and elaboration of the current legislation of Ukraine, an algorithm for forensic investigation of objects containing synthetic cannabinoids has been developed. Conclusions. The stages of the forensic analysis of objects containing synthetic cannabinoids meet the requirements of the current legislation of Ukraine and the Ministry of Justice of Ukraine. The obtained data prove the high sensitivity and reproducibility of the methods and prove the possibility of their introduction into the practice of forensic examination
... Although CBD is said to have low binding affinity to both CB 1 and CB 2 receptors [140], it interacts with CB 1 receptors by acting as a negative allosteric modulator [141]. Recent studies [142] have also shown that CBD acts as an inverse agonist of the CB 2 receptor, which could explain its anti-inflammatory properties as CB 2 receptors are expressed remarkably in immune cells, and inhibiting their migration could potentially reduce inflammation [143]. In stark contrast to THC, CBD does not exhibit psychoactive properties, which renders it useful for the treatment of various disorders. ...
Article
Full-text available
Sleep is an essential biological phase of our daily life cycle and is necessary for maintaining homeostasis, alertness, metabolism, cognition, and other key functions across the animal kingdom. Dysfunctional sleep leads to deleterious effects on health, mood, and cognition, including memory deficits and an increased risk of diabetes, stroke, and neurological disorders. Sleep is regulated by several brain neuronal circuits, neuromodulators, and neurotransmitters, where cannabinoids have been increasingly found to play a part in its modulation. Cannabinoids, a group of lipid metabolites, are regulatory molecules that bind mainly to cannabinoid receptors (CB1 and CB2). Much evidence supports the role of cannabinoid receptors in the modulation of sleep, where their alteration exhibits sleep-promoting effects, including an increase in non-rapid-eye movement sleep and a reduction in sleep latency. However, the pharmacological alteration of CB1 receptors is associated with adverse psychotropic effects, which are not exhibited in CB2 receptor alteration. Hence, selective alteration of CB2 receptors is also of clinical importance, where it could potentially be used in treating sleep disorders. Thus, it is crucial to understand the neurobiological basis of cannabinoids in sleep physiology. In this review article, the alteration of the endocannabinoid system by various cannabinoids and their respective effects on the sleep-wake cycle are discussed based on recent findings. The mechanisms of the cannabinoid receptors on sleep and wakefulness are also explored for their clinical implications and potential therapeutic use on sleep disorders.
... Cannabinoids are found in cannabis (cannabis sativa) and cannabis products (Pertwee, 2004;Elsohly et al., 2005). Cannabidiol (CBD), cannabigerol (CBG), cannabinol and cannabichromene (CBC) are other most important compounds (Williamson et al., 2000). ...
Chapter
Cannabis is a genus of flowering plants in the Cannabaceae family and generally has three species, Cannabis sativa, C. indica, and C. ruderalis. Tetrahydrocannabinol (THC) and cannabidiol are well-known cannabinoids found in cannabis. The elucidation and characterization of the structures of cannabinoids used in the treatment of various diseases is one of the most important steps. Vibrational spectroscopy, FTIR, and Raman spectroscopies are spectroscopic techniques that have been important in characterizing the molecular properties and functional groups of cannabinoids. Over the years, this technique, which analyzes samples in a fast, environmentally-friendly, and non-destructive manner has shown great success in drug analysis and has led to great technological developments accordingly. This chapter presents a historical overview of the uses of cannabinoids in medicine, explains the applications of QR codes in the field of medical cannabis, and offers examples of characteristic vibrational wavenumbers of natural and synthetic cannabinoids using FTIR and Raman spectroscopies.
... In terms of functional activity, multiple reports have shown that CBD antagonizes the agonist activity of CP 55,940 and R-(+) WIN55212 and in some cases, it behaves as an inverse agonist at CB1R depending on the used concentration. At CB2R, CBD behaved as an inverse agonist with promising anti-inflammatory properties in murine microglia and macrophages, reviewed in Pertwee (2004Pertwee ( , 2008. The synthetic pathway of Δ 9 -THC, CBD, and some other pCBs (i.e., cannabichromene [CBC]) share cannabigerol (CBG) as the main precursor that converts via specific synthase enzymes (e.g., Δ 9 -THC synthase, CBD synthase, or CBC synthase) into various pCBs. ...
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The cover image is based on the Original Article Phytocannabinoids regulate inflammation in IL‐1β‐stimulated human gingival fibroblasts by Ammaar H. Abidi et al., https://doi.org/10.1111/jre.13050
... The pharmacological attributes of THC are related to its high affinity for the type-1 cannabinoid receptor (CB 1 R) [8]. CBD interacts with both receptors of the human endocannabinoid system (ECS), CB 1 R and type-2 cannabinoid receptor (CB 2 R), although with lower affinities, compared to THC [9]. The regulatory functions carried out by ECS in the central nervous system are cognition, appetite control, and analgesia [10]. ...
Article
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Roadside testing of illicit drugs such as tetrahydrocannabinol (THC) requires simple, rapid, and cost-effective methods. The need for non-invasive detection tools has led to the development of selective and sensitive platforms, able to detect phyto- and synthetic cannabinoids by means of their main metabolites in breath, saliva, and urine samples. One may estimate the time passed from drug exposure and the frequency of use by corroborating the detection results with pharmacokinetic data. In this review, we report on the current detection methods of cannabinoids in biofluids. Fluorescent, electrochemical, colorimetric, and magnetoresistive biosensors will be briefly overviewed, putting emphasis on the affinity formats amenable to on-site screening, with possible applications in roadside testing and anti-doping control.
... Hemp primarily produces CBD, a nonpsychoactive cannabinoid that can be used for therapeutic, medicinal, and recreational purposes (Andre, Hausman, & Guerriero, 2016;World Health Organization, 2018). CBD has been used to aid ailments such as arthritis, epilepsy, inflammation, anxiety, and pain (Evans Schultes, Klein, Plowman, & Lockwood, 1974;Pertwee, 2004;Rosenberg, Tsien, Whalley, & Devinsky, 2015; World Health Organization, 2018). ...
Chapter
Hemp production has persisted over several centuries, although its relevance has ebbed and flowed. Its popularity has been interrupted by competing products, public health concerns, and regulatory barriers. A renaissance of hemp production has emerged over the past few decades, giving rise to a market of diverse hemp-based food, medicinal, textile, and industrial products. Critical to the crop’s long-term success is an understanding of the hemp consumer, without whom hemp cannot survive. This chapter discusses the renewed interest in hemp across the global market landscape and the importance of intentional communication of hemp to the consumer.
... In terms of functional activity, multiple reports have shown that CBD antagonizes the agonist activity of CP 55,940 and R-(+) WIN55212 and in some cases, it behaves as an inverse agonist at CB1R depending on the used concentration. At CB2R, CBD behaved as an inverse agonist with promising anti-inflammatory properties in murine microglia and macrophages, reviewed in Pertwee (2004Pertwee ( , 2008. The synthetic pathway of Δ 9 -THC, CBD, and some other pCBs (i.e., cannabichromene [CBC]) share cannabigerol (CBG) as the main precursor that converts via specific synthase enzymes (e.g., Δ 9 -THC synthase, CBD synthase, or CBC synthase) into various pCBs. ...
Article
Full-text available
Introduction: In the last two decades, our understanding of the therapeutic utility and medicinal properties of cannabis has greatly changed. This change has been accompanied by widespread cannabis use in various communities and different age groups, especially within the United States. With this increase, we should consider the potential effects of cannabis-hemp on general public health and how they could alter therapeutic outcomes. Material and methods: The present investigation examined cannabis use for recreational and therapeutic use and a review of pertinent indexed literature was performed. The focused question evaluates "how cannabis or hemp products impact health parameters and do they provide potential therapeutic value in dentistry, and how do they interact with conventional medicines (drugs)." Indexed databases (PubMed/Medline, EMBASE) were searched without any time restrictions but language was restricted to English. Results: The review highlights dental concerns of cannabis usage, the need to understand the endocannabinoid system (ECS), cannabinoid receptor system, its endogenous ligands, pharmacology, metabolism, current oral health, and medical dilemma to ascertain the detrimental or beneficial effects of using cannabis-hemp products. The pharmacological effects of pure cannabidiol (CBD) have been studied extensively while cannabis extracts can vary significantly and lack empirical studies. Several metabolic pathways are affected by cannabis use and could pose a potential drug interaction. The chronic use of cannabis is associated with health issues, but the therapeutic potential is multifold since there is a regulatory role of ECS in many pathologies. Conclusion: Current shortcomings in understanding the benefits of cannabis or hemp products are limited due to pharmacological and clinical effects not being predictable, while marketed products vary greatly in phytocompounds warrant further empirical investigation. Given the healthcare challenges to manage acute and chronic pain, this review highlights both cannabis and CBD-hemp extracts to help identify the therapeutic application for patient populations suffering from anxiety, inflammation, and dental pain.
... From a psychiatric perspective, CBD is a novel promising therapeutic agent. It attenuates the psychotic-like effects of cannabis over time in recreational users [154] and it was repeatedly shown to induce anxiolytic activity in preclinical and clinical studies [41, [155][156][157] in addition to its anti-compulsive effects [158][159][160][161]. Furthermore, it exhibits anti-depressivelike abilities in several animal models [42][43][44], yet, to the best of our knowledge, there is no published controlled clinical study that has investigated whether CBD can decrease depressive symptoms in patients. A laboratory study reported that CBD attenuated the transient "amotivational" effects of THC-based cannabis [162] and a small-scale clinical trial focusing on different outcomes reported that depression was an intervening variable in the effects of Sativex (which has a high ratio of CBD:THC) on the main topic of interest [163]. ...
Article
Full-text available
Depression and anxiety disorders are two of the most common and growing mental health concerns in adolescents. Consequently, antidepressant medication (AD) use has increased widely during the last decades. Several classes of antidepressants are used mainly to treat depression, anxiety, and obsessive-compulsive disorders by targeting relevant brain neurochemical pathways. Almost all randomized clinical trials of antidepressants examined patients with no concomitant medications or drugs. This does not address the expected course of therapy and outcome in cannabis users. Cannabis is the most commonly used illicit substance globally. Substantial changes in its regulation are recently taking place. Many countries and US states are becoming more permissive towards its medical and recreational use. The psychological and physiological effects of cannabis (mainly of its major components, tetrahydrocannabinol (THC) and cannabidiol (CBD)) have been extensively characterized. Cannabis use can be a risk factor for depressive and anxiety symptoms, but some constituents or mixtures may have antidepressant and/or anxiolytic potential. The aim of this literature review is to explore whether simultaneous use of AD and cannabis in adolescence can affect AD treatment outcomes. Based on the current literature, it is reasonable to assume that antidepressants are less effective for adolescents with depression/anxiety who frequently use cannabis. The mechanisms of action of antidepressants and cannabis point to several similarities and conjunctions that merit future investigation regarding the potential effectiveness of antidepressants among adolescents who consume cannabis regularly.
... Entre los más de 66 cannabinoides conocidos (Pertwee, 2004), producidos por la planta encontramos: tetrahidrocannabinol (THC), cannabidiol (CBD), cannabinol (CBN), cannabichromene (CBC), cannabigerol (CBG) y al menos 60 otros cannabinoides. ...
Technical Report
Full-text available
En este trabajo se describe el valor agregado de los cultivos agroindustriales de fibras vegetales, como el junco, algodón, lino, cáñamo y paste, además se analiza las condiciones agroecológicas, producción, y transformación de estas materias primas.
... Nowadays, several recovery approaches have been used to isolate cannabinoid compounds from its plant-based matrix. Extraction has played a vital role in the production of medicines; therefore, it is crucial to the development of cannabis-based medicinal products [10][11][12]. Conventional extraction techniques were found to have adverse toxic effects on the environment and human health [13][14][15]. Therefore, it is important to develop "green" extraction techniques for efficient extraction of the desired cannabinoids from the cannabis plant. ...
Article
Full-text available
Isolation of the therapeutic cannabinoid compounds from Cannabis Sativa L. (C. Sativa) is important for the development of cannabis-based pharmaceuticals for cancer treatment, among other ailments. The main pharmacological cannabinoids are THC and CBD. However, THC also induces undesirable psychoactive effects. The decarboxylation process converts the naturally occurring acidic forms of cannabinoids, such as cannabidiolic acid (CBDA) and tetrahydrocannabinolic acid (THCA), to their more active neutral forms, known as cannabidiol (CBD) and tetrahydrocannabinol (THC). The purpose of this study was to selectively extract cannabinoids using a novel in situ decarboxylation pressurized hot water extraction (PHWE) system. The decarboxylation step was evaluated at different temperature (80–150 °C) and time (5–60 min) settings to obtain the optimal conditions for the decarboxylation-PHWE system using response surface methodology (RSM). The system was optimized to produce cannabis extracts with high CBD content, while suppressing the THC and CBN content. The identification and quantification of cannabinoid compounds were determined using UHPLC-MS/MS with external calibration. As a result, the RSM has shown good predictive capability with a p-value < 0.05, and the chosen parameters revealed to have a significant effect on the CBD, CBN and THC content. The optimal decarboxylation conditions for an extract richer in CBD than THC were set at 149.9 °C and 42 min as decarboxylation temperature and decarboxylation time, respectively. The extraction recoveries ranged between 96.56 and 103.42%, 95.22 and 99.95%, 99.62 and 99.81% for CBD, CBN and THC, respectively.
... When heated, THCA and CBDA are converted into the neutral forms THC and CBD (Russo, 2011), which interact with the human endocannabinoid system (Pertwee, 1988(Pertwee, , 1997(Pertwee, , 2004. Both THC and CBD have medicinal (Russo, 2011;Swift et al., 2013;Volkow et al., 2014) and economic value (Evans, 2013;Kirsch, 2018), but THC has been intensely selected by breeders and growers (Volkow et al., 2014) due to its psychoactive effects (ElSohly & Slade, 2005). ...
Article
Full-text available
The flowering plant Cannabis sativa , cultivated for centuries for multiple purposes, displays extensive variation in phenotypic traits in addition to its wide array of secondary metabolite production. Notably, Cannabis produces two well-known secondary-metabolite cannabinoids: cannabidiolic acid (CBDA) and delta-9-tetrahydrocannabinolic acid (THCA), which are the main products sought by consumers in the medical and recreational market. Cannabis has several suggested subspecies which have been shown to differ in chemistry, branching patterns, leaf morphology and other traits. In this study we obtained measurements related to phytochemistry, reproductive traits, growth architecture, and leaf morphology from 297 hybrid individuals from a cross between two diverse lineages. We explored correlations among these characteristics to inform our understanding of which traits may be causally associated. Many of the traits widely assumed to be strongly correlated did not show any relationship in this hybrid population. The current taxonomy and legal regulation within Cannabis is based on phenotypic and chemical characteristics. However, we find these traits are not associated when lineages are inter-crossed, which is a common breeding practice and forms the basis of most modern marijuana and hemp germplasms. Our results suggest naming conventions based on leaf morphology do not correspond to the chemical properties in plants with hybrid ancestry. Therefore, a new system for identifying variation within Cannabis is warranted that will provide reliable identifiers of the properties important for recreational and, especially, medical use.
... 17−20 Nowadays the most commercially trending component of cannabis is CBD, since its psychoactivity is not the same as of THC-9. 21,22 Moreover, due to the yearly increase of cases of neurodegenerative diseases, 23−25 CBD becomes a very attractive drug because it has already shown the great potential against them in various experimental studies. 26 31 showed on rat primary astroglial cultures that CBD could reduce the inflammation which was Aβ-induced. ...
Article
In this work cannabidiol (CBD) was investigated as a possible drug against the cytotoxicity of Aβ(31−35) and Aβ(25− 35) peptides with the help of atomistic molecular dynamics (MD) and well-tempered metadynamics simulations. Four interrelated mechanisms of possible actions of CBD are proposed from our computations. This implies that one mechanism can be a cause or/ and a consequence of another. CBD is able to decrease the aggregation of peptides at certain concentrations of compounds in water. This particular action is more prominent for Aβ(25−35), since originally Aβ(31−35) did not exhibit aggregation properties in aqueous solutions. Interactions of CBD with the peptides affect secondary structures of the latter ones. Clusters of CBD are seen as possible adsorbents of Aβ(31−35) and Aβ(25−35) since peptides are tending to aggregate around them. And last but not least, CBD exhibits binding to MET 35. All four mechanisms of actions can possibly inhibit the Aβ-cytotoxicity as discussed in this paper. Moreover, the amount of water also played a role in peptide clustering: with a growing concentration of peptides in water without a drug, the aggregation of both Aβ(31−35) and Aβ(25−35) increased. The number of hydrogen bonds between peptides and water was significantly higher for simulations with Aβ(25−35) at the higher concentration of peptides, while for Aβ(31−35) that difference was rather insignificant. The presence of CBD did not substantially affect the number of hydrogen bonds in the simulated systems.
... Cannabidiol (CBD) and Δ 9 -tetrahydrocannabinol (Δ 9 -THC) are the main cannabinoids with medicinal potential present in plants of the genus Cannabis [1][2][3]. However, the highest concentrations of cannabinoids in fresh flowers are cannabidiolic acid (CBDA) and Δ 9 -tetrahydrocannabinoic acid (Δ 9 -THCA). ...
Article
The cannabidiol (CBD) and δ⁹-tetrahydrocannabinol (δ⁹-THC) are the two main cannabinoids with medicinal potential. However, these substances are found in low concentrations in fresh Cannabis flowers. The decarboxylation process can be applied to fresh flowers to convert the cannabinoids acids present in high contents into neutral cannabinoids of interest (CBD and δ⁹-THC). The purpose of this work is to present the results of two strategies of cannabinoid extraction with supercritical carbon dioxide (scCO2) using different varieties of flowers of the genus Cannabis. The first strategy was the process of decarboxylation of flowers followed by extraction with scCO2. In the second strategy, the process of decarboxylation of the flowers was not employed, and the extraction was conducted using scCO2 with 6 % ethanol as a co-solvent. The purification process called "winterization" was also analyzed. The chemical profiles of neutral cannabinoids (CBD, δ⁹-THC and cannabinol (CBN)) were analyzed by high-performance liquid chromatography with photodiode array detection (HPLC/ PDA) and of essential oils from the fresh flowers extracts obtained employing the second strategy and using a mixture of organic solvents were analyzed by gas chromatography with mass spectrometry detection (GC/MS). The application of the decarboxylation process before extractions potentiated the CBD and δ⁹-THC content of flower samples on 5 to 10 fold. Although the use of second strategy presented extracts with essential oils and high yields (30 %) the content of cannabinoid in extracts was low (< 15 %). The first extraction strategy presented extracts with yield up to 25 % with similar high contents of CBD and δ⁹-THC (35 % each) or only high contents of ?9-THC between 77 and 88 %.
... Despite these positive effects, there is a serious lack of carefully controlled clinical research in the field. protective properties in several animal models of neurodegeneration as well as therapeutic-like effects in models of psychiatric disorders [19,29,35,46,47]. Clinical trials have indicated potential benefits in the management of Alzheimer's disease, multiple sclerosis (MS), Parkinson's disease and amyotrophic lateral sclerosis [18]. ...
Article
The current study explores the therapeutic potential of Cannabidiol (CBD), a compound in the Cannabis plant, using both sexes of 2 “depressive-like” genetic models, Wistar Kyoto (WKY) and Flinders Sensitive Line (FSL) rats. Rats ingested CBD (30 mg/kg) orally. In the saccharin preference test, following a previous report of a pro-hedonic effect of CBD in male WKY, we now found similar results in female WKY. CBD also decreased immobility in the forced swim test in males (both strains) and in female WKY. These findings suggest a role for CBD in treating mental disorders with prominent symptoms of helplessness and anhedonia.
... Traditional ''cannabinoids'' or ''phytocannabinoids'' include at least 66 biologically active terpene phenolic compounds, derivatives of two-substituted 5-amylresorcine, that are found in Cannabis sativa herb 1,2 or are the products thereof such as herbal cannabis (''marihuana''), cannabis resin (''hashish''), and liquid cannabis (''cannabis oil''). 3 Their synthetic analogues capable of binding to cannabinoid receptors 4,5 are also generally referred to as ''cannabinoids.'' ...
Article
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Synthetic cannabinoids are one of the most rapidly expanding classes of novel psychoactive substances found in illegal markets. These substances have evolved to the point that many examples no longer fit with the traditional cannabinoid classification system, where assignment of these compounds is difficult and ambiguous, leading to inconsistencies in regard to their chemical structures. This and other drawbacks can result in misunderstandings between forensic scientists and legal disciplines, complicating efforts toward improving the inadequacies of current antidrug laws. After a critical review, we offer an updated yet simplified cannabinoid classification system with the intention to facilitate interdisciplinary communication.
... CBD, however, produces antiepileptiform and anticonvulsant effects in both in vitro and in vivo models. [64,51,[61][62][63]85] More recently in 1980, Cunha et al., published a double-blind study that evaluated CBD for intractable epilepsy in 16 patients with grand-mal seizures. Each patient received 200-300 mg daily of CBD or placebo along with antiepileptic drugs for up to 4 months. ...
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Background Numerous physical, psychological, and emotional benefits have been attributed to marijuana since its first reported use in 2,600 BC in a Chinese pharmacopoeia. The phytocannabinoids, cannabidiol (CBD), and delta-9-tetrahydrocannabinol (Δ9-THC) are the most studied extracts from cannabis sativa subspecies hemp and marijuana. CBD and Δ9-THC interact uniquely with the endocannabinoid system (ECS). Through direct and indirect actions, intrinsic endocannabinoids and plant-based phytocannabinoids modulate and influence a variety of physiological systems influenced by the ECS. Methods In 1980, Cunha et al. reported anticonvulsant benefits in 7/8 subjects with medically uncontrolled epilepsy using marijuana extracts in a phase I clinical trial. Since then neurological applications have been the major focus of renewed research using medical marijuana and phytocannabinoid extracts. Results Recent neurological uses include adjunctive treatment for malignant brain tumors, Parkinson's disease, Alzheimer's disease, multiple sclerosis, neuropathic pain, and the childhood seizure disorders Lennox-Gastaut and Dravet syndromes. In addition, psychiatric and mood disorders, such as schizophrenia, anxiety, depression, addiction, postconcussion syndrome, and posttraumatic stress disorders are being studied using phytocannabinoids. Conclusions In this review we will provide animal and human research data on the current clinical neurological uses for CBD individually and in combination with Δ9-THC. We will emphasize the neuroprotective, antiinflammatory, and immunomodulatory benefits of phytocannabinoids and their applications in various clinical syndromes.
... Both D 8 THC and D 9 THC produce antinociceptive effects in pre-clinical models with similar potency via activation of CB 1 R. [22][23][24][25][26] CBD lacks the behavioral effects of THC at CB 1 R, and may produce pharmacological actions through the activation of noncannabinoid receptors. [27][28][29] HU-308 is a selective and highly potent agonist at CB 2 R, 30 and has previously been shown to reduce lipopolysaccharideinduced intraocular inflammation. 19 ...
Article
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Background and Purpose: Corneal injury can result in dysfunction of corneal nociceptive signaling and corneal sensitization. Activation of the endocannabinoid system has been reported to be analgesic and anti-inflammatory. The purpose of this research was to investigate the antinociceptive and anti-inflammatory effects of cannabinoids with reported actions at cannabinoid 1 (CB1R) and cannabinoid 2 (CB2R) receptors and/or noncannabinoid receptors in an experimental model of corneal hyperalgesia. Methods: Corneal hyperalgesia (increased pain response) was generated using chemical cauterization of the corneal epithelium in wild-type (WT) and CB2R knockout (CB2R−/−) mice. Cauterized eyes were treated topically with the phytocannabinoids Δ⁸-tetrahydrocannabinol (Δ⁸THC) or cannabidiol (CBD), or the CBD derivative HU-308, in the presence or absence of the CB1R antagonist AM251 (2.0 mg/kg i.p.), or the 5-HT1A receptor antagonist WAY100635 (1 mg/kg i.p.). Behavioral pain responses to a topical capsaicin challenge at 6 h postinjury were quantified from video recordings. Mice were euthanized at 6 and 12 h postcorneal injury for immunohistochemical analysis to quantify corneal neutrophil infiltration. Results: Corneal cauterization resulted in hyperalgesia to capsaicin at 6 h postinjury compared to sham control eyes. Neutrophil infiltration, indicative of inflammation, was apparent at 6 and 12 h postinjury in WT mice. Application of Δ⁸THC, CBD, and HU-308 reduced the pain score and neutrophil infiltration in WT mice. The antinociceptive and anti-inflammatory actions of Δ⁸THC, but not CBD, were blocked by the CB1R antagonist AM251, but were still apparent, for both cannabinoids, in CB2R−/− mice. However, the antinociceptive and anti-inflammatory actions of HU-308 were absent in the CB2R−/− mice. The antinociceptive and anti-inflammatory effects of CBD were blocked by the 5-HT1A antagonist WAY100635. Conclusion: Topical cannabinoids reduce corneal hyperalgesia and inflammation. The antinociceptive and anti-inflammatory effects of Δ⁸THC are mediated primarily via CB1R, whereas that of the cannabinoids CBD and HU-308, involve activation of 5-HT1A receptors and CB2Rs, respectively. Cannabinoids could be a novel clinical therapy for corneal pain and inflammation resulting from ocular surface injury.
... Despite these positive effects, there is a serious lack of carefully controlled clinical research in the field. protective properties in several animal models of neurodegeneration as well as therapeutic-like effects in models of psychiatric disorders [19,29,35,46,47]. Clinical trials have indicated potential benefits in the management of Alzheimer's disease, multiple sclerosis (MS), Parkinson's disease and amyotrophic lateral sclerosis [18]. ...
... Therefore, the primary aim of our work was to assess whether the concentrations of CBD and THC found in the intestinal lymphatic system following oral co-administration with lipids could reach levels that are sufficiently high to produce immunomodulatory effects. MS and cancer patients on chemotherapy regimen were selected in this study as model cases of autoimmune illness and immunocompromised status, respectively, based on the fact that cannabinoids are currently used as symptomatic treatment in both patients groups 25, 26 , and have immunomodulatory activity 1 . Therefore, an additional aim was to assess if such high levels in the intestinal lymphatic system are of potential therapeutic value to improve the treatment outcomes of autoimmune diseases such as MS, or can lead to potential safety considerations in immunocompromised patients such as those under chemotherapy regimens. ...
Article
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Cannabidiol (CBD) and Δ9-tetrahydrocannabinol (THC) have well documented immunomodulatory effects in vitro, but not following oral administration in humans. Here we show that oral co-administration of cannabinoids with lipids can substantially increase their intestinal lymphatic transport in rats. CBD concentrations in the lymph were 250-fold higher than in plasma, while THC concentrations in the lymph were 100-fold higher than in plasma. Since cannabinoids are currently in clinical use for the treatment of spasticity in multiple sclerosis (MS) patients and to alleviate nausea and vomiting associated with chemotherapy in cancer patients, lymphocytes from those patients were used to assess the immunomodulatory effects of cannabinoids. The levels of cannabinoids recovered in the intestinal lymphatic system, but not in plasma, were substantially above the immunomodulatory threshold in murine and human lymphocytes. CBD showed higher immunosuppressive effects than THC. Moreover, immune cells from MS patients were more susceptible to the immunosuppressive effects of cannabinoids than those from healthy volunteers or cancer patients. Therefore, administering cannabinoids with a high-fat meal or in lipid-based formulations has the potential to be a therapeutic approach to improve the treatment of MS, or indeed other autoimmune disorders. However, intestinal lymphatic transport of cannabinoids in immunocompromised patients requires caution.
... Cannabidiol was first isolated from Cannabis in the late 1930s [51], and has since been found to have therapeutic potential for disorders such as inflammation and anxiety, and also has potential as a neuroprotective agent and an antioxidant [52,53]. Along with CBD, other minor cannabinoids such as cannabidiolic acid (CBDA), tetrahydrocannabinolic acid (THCA), and cannabidivarin (CBDV) have shown efficacy for relief of nausea and emesis [54]. Cannabidiol also has been used to treat arthritis, cancer, diabetes, neurodegenerative diseases and pain [23]. ...
Article
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Most of the Western World banned the cultivation of Cannabis sativa in the early 20th century because biotypes high in ∆9-tetrahydrocannabinol (THC, the principal intoxicant cannabinoid) are the source of marijuana. Nevertheless, since 1990, dozens of countries have authorized the licensed growth and processing of “industrial hemp” (cultivars with quite low levels of THC). Canada has concentrated on hemp oilseed production, and very recently, Europe changed its emphasis from fiber to oilseed. The USA, historically a major hemp producer, appears on the verge of reintroducing industrial hemp production. This presentation provides updates on various agricultural, scientific, social, and political considerations that impact the commercial hemp industry in the United States and Canada. The most promising scenario for the hemp industry in North America is a continuing focus on oilseed production, as well as cannabidiol (CBD), the principal non-intoxicant cannabinoid considered by many to have substantial medical potential, and currently in great demand as a pharmaceutical. Future success of the industrial hemp industry in North America is heavily dependent on the breeding of more productive oilseed cultivars, the continued development of consumer goods, reasonable but not overly restrictive regulations, and discouragement of overproduction associated with unrealistic enthusiasm. Changing attitudes have generated an unprecedented demand for the cannabis plant and its products, resulting in urgent needs for new legislative, regulatory, and business frameworks, as well as scientific, technological, and agricultural research.
... The two main natural cannabinoids, the psychoactive ∆ 9 -tetrahydrocannabinol (THC) and the non-psychoactive cannabidiol (CBD), have been the focus of research over the last few decades for their potential multiple therapeutic effects [14]. Both cannabinoids are currently available as pharmaceutical formulations. ...
Article
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There has been an escalating interest in the medicinal use of Cannabis sativa in recent years. Cannabis is often administered orally with fat-containing foods, or in lipid-based pharmaceutical preparations. However, the impact of lipids on the exposure of patients to cannabis components has not been explored. Therefore, the aim of this study is to elucidate the effect of oral co-administration of lipids on the exposure to two main active cannabinoids, ∆9-tetrahydrocannabinol (THC) and cannabidiol (CBD). In this study, oral co-administration of lipids enhanced the systemic exposure of rats to THC and CBD by 2.5-fold and 3-fold, respectively, compared to lipid-free formulations. In vitro lipolysis was conducted to explore the effect of lipids on the intestinal solubilisation of cannabinoids. More than 30% of THC and CBD were distributed into micellar fraction following lipolysis, suggesting that at least one-third of the administered dose will be available for absorption following co-administration with lipids. Both cannabinoids showed very high affinity for artificial CM-like particles, as well as for rat and human CM, suggesting high potential for intestinal lymphatic transport. Moreover, comparable affinity of cannabinoids for rat and human CM suggests that similar increased exposure effects may be expected in humans. In conclusion, co-administration of dietary lipids or pharmaceutical lipid excipients has the potential to substantially increase the exposure to orally administered cannabis and cannabis-based medicines. The increase in patient exposure to cannabinoids is of high clinical importance as it could affect the therapeutic effect, but also toxicity, of orally administered cannabis or cannabis-based medicines.
... D 9 -THC likely acts mainly by activating CB 1 receptors on presynaptic terminals of glutamatergic and GABAergic synapses and suppressing the presynaptic release of these neurotransmitters [60], while CBD likely acts by interacting with 5-HT 1 a receptors leading to an increased cerebral blood flow and neuroprotective effect [61]. Cannabinoids, particularly CBD, have been proposed for treatment of spasticity and pain symptoms associated with multiple sclerosis and amyotrophic lateral sclerosis, and some CBD analogs have been proposed for neuroprotection in glaucoma to delay the progressive damage of the optic nerve [62]. ...
Chapter
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Cannabinoids, a class of meroterpenoids derived from the alkylation of an olivetol-like alkyl resorcinol with a monoterpene unit, are the most typical constituents of Cannabis. This class includes over a hundred members belonging to several structural types, mainly differing by the constitution of their terpenoid moiety. The biomedical relevance of cannabinoids transcends the psychotropic properties of Δ9-THC since, because of their anti-inflammatory, analgesic, and antibacterial activity, they show potential in various fields of medicine, addressing unmet needs like the symptomatic mitigation of multiple sclerosis and the treatment of MRSA infections. Research on cannabinoid activities has yielded to the discovery of an entire new class of human receptors, called cannabinoid (CB) receptors, and of their endogenous agonists, collectively named endocannabinoids (EC). Since its discovery in the early 1990s, the EC system has increasingly emerged as a key signaling system involved in a plethora of physiological and pathological functions in mammals. This chapter will provide an overview on this multi-faceted class of natural products.
... Synthetic chemists make much effort to design compounds that would be highly selective to CB 2 receptors but would have no psychoactive properties. To this end, various carb oxamides were synthesized, including those based on py ridine, 2 oxo 1,2 dihydropyridine, 122 5 arylisoxazole, 123 benzimidazole, 124 7 oxo [1,4]oxazino [2,3,4 ij]quino line, 125 biphenyl, 126 and tricyclic pyrazole structures. 127 1,3,5 Triazine derivatives 128 and 3 arylcarboxamide de rivatives of 1,2 dihydro 2 oxopyridine, 129 5 , 6 , and 7 azaindole, 130,131 and thiazole 132 were tested for activity at CB 2 receptors. ...
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The data on the chemical structures, biological effects, and use of cannabinoids on the illegal market of new psychoactive substances were generalized. An extended classification comprising new classes, groups, and subgroups of cannabinoids was proposed for better repre� sentation of their structural variety. The emergence of new synthetic cannabinoids which belong to the groups of cycloalkanecarbonylindoles, indole� and indazole�3�carboxamides, and indole� and indazole�3�carboxylates is closely associated with the market of new psycho� active substances
... In addition to the above-mentioned therapeutic applications already in clinical use, efficacy studies have shown that both CBD and THC have a broad spectrum of other therapeutic activities [13][14][15][16]. A significant number of these studies have been conducted in animal models of the diseases in question, such as multiple sclerosis [17], glaucoma [18], acute colitis [19], epilepsy [20][21][22][23][24], oedema and hyperalgesia [25], anxiety [26,27], and psychosis [28]. ...
Article
There has been increased interest in the medical use of cannabinoids in recent years, particularly in the predominant natural cannabinoids, cannabidiol (CBD) and Δ(9)-tetrahydrocannabinol (THC). The aim of the current study was to develop a sensitive and reliable method for the quantification of CBD and THC in rat plasma. A combination of protein precipitation using cold acetonitrile and liquid-liquid extraction using n-hexane was utilised to extract CBD and THC from rat plasma. Samples were then evaporated and reconstituted in acetonitrile and 30μL was injected into an HPLC system. Separation was achieved using an ACE C18-PFP 150mm×4.6mm, 3μm column at 55°C with isocratic elution using a mobile phase consisting of acetonitrile-water (62:38, v/v) at 1mL/min for 20min. Both cannabinoids, as well as the internal standard (4,4-dichlorodiphenyltrichloroethane, DDT) were detected at 220nm. Our new method showed linearity in the range of 10-10,000ng/mL and a lower limit of quantification (LLOQ) of 10ng/mL for both cannabinoids, which is comparable to previously reported LC-MS/MS methods. Inter- and intra-day precision and accuracy were below 15% RSD and RE, respectively. To demonstrate the suitability of the method for in vivo studies in rats, the assay was applied to a preliminary pharmacokinetic study following IV bolus administration of 5mg/kg CBD or THC. In conclusion, a simple, sensitive, and cost-efficient HPLC-UV method for the simultaneous determination of CBD and THC has been successfully developed, validated and applied to a pharmacokinetic study in rats. Copyright © 2015 Elsevier B.V. All rights reserved.
... It is used by men for a long time. Hemp has been used for fibre (textile and paper industry, engineering), for seed and seed oils (food, engineering), for medicinal purposes, and as a soft drug [1][2][3]. "Industrial hemp" is represented by species of Cannabis sativa. Its used varieties have been developed to contain minimal levels of THC (terpeno-phenolic compound: tetrahydrocannabinol). ...
Conference Paper
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This experiment was focus to increase the germination and growth of hemp seed by using cold plasma discharge. The seeds of three hemp cultivars (Finola, Bialobrzeskie, and Carmagnola) were pre-treated in Plasonic AR-550-M and GlidArc apparatuses for three time expositions (180 s, 300 s, and 600 s). Commercial Plasonic AR-550-M apparatus was not suitable for plasma pre-treatment, because all seeds were obviously under a big stress. GlidArc device was suitable. GlidArc plasma pre-treatment positively affected the seeds of the cultivar Finola in all tested expositions above all. Seedling acceleration on the fifth day of the experiment showed higher values than control sample (153% in length and 127% in weight of seedlings). Although tested cultivars were taxonomically very similar, high differences in response to plasma pre-treatment were found.
... This may be attributed to the fact that CBD has much lower affinity for CB 1 and CB 2 receptors than Δ 9 -THC. 56 Ursolic acid, a pentacyclic triterpenoid that is widely distributed in plants, has been discovered to have anticonvulsant activity because it reduced the number and lethality of PTZ-induced seizures in mice. 57 Ursolic acid stearoyl glucoside, a terpenoid isolated from Lantana camara L., also exhibited a anticonvulsive effect. ...
Article
Objective Epilepsy is a serious neural disease that affects around 50million people all over the world. Although for the majority patients with epilepsy, seizures are well controlled by currently available antiepileptic drugs (AEDs), there are still >30% of patients suffered from medically refractory epilepsy and approximately 30-40% of all epileptic patients affected by numerous side effects and seizure resistance to the current AEDs. Therefore, many researchers try to develop novel approaches to treat epilepsy, for example, to discover new antiepileptic constituents from herbal medicines. Although there are already several reviews on phytotherapy in epilepsy, most of them placed emphasis on the plant crude extracts or their isolated fractions, not pure active compounds derived from herbal medicines. This article aims to review components in herbal medicines that have shown antiepileptic or anticonvulsant properties. Methods We searched online databases and identified articles using the preset searching syntax and inclusion criteria. The active medicinal compounds that have shown anticonvulsant or antiepileptic activity were included and classified according to structural types. ResultsWe have reviewed herein the active constituents including alkaloids, flavonoids, terpenoids, saponins, and coumarins. The screening models, the seizures-inducing factors and response, the effective dose, the potential mechanisms, as well as the structure-activity relationships in some of these active components have also been discussed. SignificanceThe in vitro and in vivo experimental data reviewed in this paper would supply the basic science evidence for research and development of novel AEDs from medicinal plants.
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An increasing number of countries have started to decriminalize or legalize the consumption of cannabis for recreational and medical purposes. The active ingredients in cannabis, termed cannabinoids, affect multiple functions in the human body, including coordination, motor skills, memory, response time to external stimuli, and even judgment. Cannabinoids are a unique class of terpeno-phenolic compounds, with 120 molecules discovered so far. There are certain situations when people under the influence of cannabis may be a risk to themselves or the public safety. Over the past two decades, there has been a growing research interest in detecting cannabinoids from various biological matrices. There is a need to develop a rapid, accurate, and reliable method of detecting cannabinoids in oral fluid as it can reveal the recent intake in comparison with urine specimens, which only show a history of consumption. Significant improvements are continuously made in the analytical formats of various technologies, mainly concerning improving their sensitivity, miniatur-ization, and making them more user-friendly. Additionally, sample collection and pretreatment have been extensively studied, and specific devices for collecting oral fluid specimens have been perfected to allow rapid and effective sample collection. This review presents the recent findings regarding the use of oral fluid specimens as the preferred biological matrix for cannabinoid detection in a point-of-care biosensor diagnostic device. A critical review is presented, discussing the findings from a collection of review and research articles, as well as publicly available data from companies that manufacture oral fluid screening devices. Firstly, the various conventional methods used to detect cannabinoids in biological matrices are presented. Secondly, the detection of cannabinoids using point-of-care biosensors is discussed, emphasizing oral fluid specimens. This review presents the current pressing technological challenges and highlights the gaps where new technological solutions can be implemented.
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Introduction Synthetic cannabinoid receptor agonists (SCRAs) are a diverse class of new psychoactive substances that have been associated with multiple instances and types of toxicity. Some SCRAs appear to carry a greater toxicological burden than others, or compared to the prototypical cannabis-derived agonist Δ⁹-tetrahydrocannabinol (Δ⁹-THC), despite a common primary mechanism of action via cannabinoid 1 (CB1) receptors. “Off-target” (i.e., non-CB1 receptor) effects could underpin this differential toxicity, although there are limited data around the activity of SCRAs at such targets. Methods A selection of 7 SCRAs (AMB-FUBINACA, XLR11, PB-22, AKB-48, AB-CHMINICA, CUMYL-PINACA, and 4F-MDMB-BUTINACA), representing several distinct chemotypes and toxicological profiles, underwent a 30 μM single-point screen against 241 G protein-coupled receptor (GPCR) targets in antagonist and agonist mode using a cellular β-arrestin recruitment assay. Strong screening “hits” at specific GPCRs were followed up in detail using concentration-response assays with AMB-FUBINACA, a SCRA with a particularly notable history of toxicological liability. Results The single-point screen yielded few hits in agonist mode for any compound aside from CB1 and CB2 receptors, but many hits in antagonist mode, including a range of chemokine receptors, the oxytocin receptor, and histamine receptors. Concentration-response experiments showed that AMB-FUBINACA inhibited most off-targets only at the highest 30 μM concentration, with inhibition of only a small subset of targets, including H1 histamine and α2B adrenergic receptors, at lower concentrations (≥1 μM). AMB-FUBINACA also produced concentration-dependent CB1 receptor signaling disruption at concentrations higher than 1 μM, but did not produce overt cytotoxicity beyond CP55,940 or Δ⁹-THC in CB1 expressing cells. Discussion These results suggest that while some “off-targets” could possibly contribute to the SCRA toxidrome, particularly at high concentrations, CB1-mediated cellular dysfunction provides support for hypotheses concerning on-target, rather than off-target, toxicity. Further investigation of non-GPCR off-targets is warranted.
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This Miniperspective of the published essential medicinal chemistry of cannabidiol (CBD) provides evidence that the popularization of CBD-fortified or CBD-labelled health products, and associated health claims, lack a rigorous scientific foundation. CBD's reputation as a cure-all puts it in the same class as other “natural” panaceas, where valid ethnobotanicals are reduced to single, purportedly active ingredients. Such reductionist approaches oversimplify useful, chemically complex mixtures in an attempt to rationalize the commercial utility of natural compounds and exploit the “natural” label. Literature evidence associates CBD with certain semi-ubiquitous, broadly-screened, primarily plant-based substances of undocumented purity that interfere with bioassays and have a low likelihood of becoming therapeutic agents. Widespread health challenges and pandemic crises such as SARS-CoV-2 create circumstances under which scientists must be particularly vigilant about healing claims that lack solid foundational data. Herein, we offer a critical review of the published medicinal chemistry properties of CBD, as well as precise definitions of CBD-containing substances and products, distilled to reveal the essential factors that impact its development as a therapeutic agent.
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Cannabis (Cannabis sativa L.), commonly known as hemp, is an annual herb belongs to the family Cannabacea. The seeds of this plant have considerable content of oil and unsaturated fatty acids, and its fiber is used in the paper and textile industries. Tetrahydrocannabinol and cannabidiol are main cannabinoid compounds of this plant, which have high importance for their well-known pharmaceutical properties. Therapeutic effects of secondary metabolites of hemp on different diseases, such as cancer, Multiple Sclerosis (M.S.), and AIDS and their anxiety and palliative characteristics have been reported in several studies. Considering oil content, and therapeutic and industrial properties of the hemp as well as, its high diversity in Iran, more studies are needed to better recognize this plant and the economic production of its therapeutic compounds. In the present paper, a comprehensive review of agronomic, therapeutic and phytochemical characteristics of hemp is presented.
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The endocannabinoid system (ECS) has emerged in recent years as a potential treatment target for alcohol use disorders (AUD). In particular, the nonpsychoactive cannabinoid cannabidiol (CBD) has shown preclinical promise in ameliorating numerous clinical symptoms of AUD. There are several proposed mechanism(s) through which cannabinoids (and CBD in particular) may confer beneficial effects in the context of AUD. First, CBD may directly impact specific brain mechanisms underlying AUD to influence alcohol consumption and the clinical features of AUD. Second, CBD may influence AUD symptoms through its actions across the digestive, immune, and central nervous systems, collectively known as the microbiota–gut–brain axis (MGBA). Notably, emerging work suggests that alcohol and cannabinoids exert opposing effects on the MGBA. Alcohol is linked to immune dysfunction (e.g., chronic systemic inflammation in the brain and periphery) as well as disturbances in gut microbial species (microbiota) and increased intestinal permeability. These MGBA disruptions have been associated with AUD symptoms such as craving and impaired cognitive control. Conversely, existing preclinical data suggest that cannabinoids may confer beneficial effects on the gastrointestinal and immune system, such as reducing intestinal permeability, regulating gut bacteria, and reducing inflammation. Thus, cannabinoids may exert AUD harm‐reduction effects, at least in part, through their beneficial actions across the MGBA. This review will provide a brief introduction to the ECS and the MGBA, discuss the effects of cannabinoids (particularly CBD) and alcohol in the brain, gut, and immune system (i.e., across the MGBA), and put forth a theoretical framework to inform future research questions.
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The inhibitory effects of Cannabis flower extracts, obtained by supercritical carbon dioxide (scCO 2 ) with and without modifier, on various human tumour cells and non-tumour cells were evaluated. Different techniques were used to optimize the polarity interaction between solute and solvent before extraction. An increase in the cannabinoid content of interest (CBD/THC) was evaluated by decarboxylation of the flowers at 110 and 140 °C. Extractions with pure scCO 2 were conducted in a single step at temperatures of 50 and 70 °C and pressures of 22 and 40 MPa, varying the time of prior decarboxylation by 0.5 or 2.0 h. Sequential extractions were conducted in a first step at 35 °C and 10 MPa and then in a second step at 70 °C and 40 MPa, with and without modifier. Contents of the cannabinoids were evaluated using HPLC. Antitumor activity of the extracts was evaluated using the MTT assay. The highest yields and highest cannabinoid contents in the extracts were obtained with high solvent density values. Extracts with high concentrations of neutral cannabinoids showed high antitumor activity for cervical cancer cell lines.
Chapter
Cannabis sativa is a multi-use crop valued for its pharmacological properties and as a fibre and seed crop. Biotechnological applications toward Cannabis research and product development are still in their early stages. An important feature of biotechnology is the collection of gene transfer technologies that are used to introduce genetic material into host organisms. Agrobacterium tumefaciens and A. rhizogenes represent the most common vectors to transfer genetic material into plant cells. Stable and transient gene expression can be achieved using A. tumefaciens while A. rhizogenes generates stable transformed hairy roots. Cannabis is amenable to genetic transformation using both Agrobacterium vectors, however the plant is recalcitrant to regeneration, impeding the recovery of transgenic Cannabis plants. Despite this shortcoming, the cannabinoid pathway is currently attracting considerable attention from the biotechnology community. Gene transfer technologies have assisted with the characterization of the cannabinoid pathway leading to the synthesis of THCA, the psychoactive compound that is highly valued as a therapeutic. Elucidation of the cannabinoid pathway has led to its metabolic engineering in heterologous hosts. The yeast Pichia pastoris has proven to be a particularly suitable host for the production of cannabinoids. Recently, biotechnology companies have emerged that anticipate commercializing cannabinoid-based drugs in yeast and tobacco and to produce hemp cultivars with the cannabinoid pathway down-regulated or completely knocked out.
Chapter
The plant Cannabis sativa has been widely used by humans over many centuries as a source of fibre, for medicinal purposes, for religious ceremonies and as a recreational drug. Since the discovery of its main psychoactive ingredient, Δ9-tetrahydrocannabinol (THC), significant progress has been made towards the understanding (1) of the in vitro and in vivo pharmacology both of THC and of certain other cannabis-derived compounds, and (2) of the potential and actual uses of these “phytocannabinoids” as medicines. There is now extensive evidence that the pharmacological effects of some widely-studied phytocannabinoids, are due to their ability to interact with cannabinoid receptors and/or with other kinds of pharmacological targets, including non-cannabinoid receptors, and this makes the pharmacology of the phytocannabinoids rather complex and interesting. In this chapter, we provide an overview of the in vitro pharmacology of five selected phytocannabinoids and report findings that have identified potential new therapeutic uses for these compounds.
Chapter
In addition to fluid haemostasis and lipid absorption, the lymphatic system and lymphoid tissues serve as the major host of immune cells where immune responses are evoked. Impaired function of the immune system might lead to serious diseases which are often treated by immunomodulators. This chapter briefly explores the physiology of an important part of the lymphatic system, the gut-associated lymphoid tissues (GALT). Currently used strategies for targeting GALT by immunomodulators for enhanced activity and/or decreased side effects are discussed. Strategies range from simple oral co-administration of immunomodulators with lipids to more advanced lipid-based formulations, polymer-based nanoparticle formulations and prodrugs. These targeting approaches successfully increase the concentration of immunomodulators achieved in the GALT and, more importantly, enhance immunomodulatory effects. Therefore, targeting immunomodulators to GALT represent a promising approach in the treatment of diseases where the immune system is actively involved.
Patent
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The present invention overcomes the problems associated with existing drug delivery systems by delivering cannabinoids transdermally. Preferably, the cannabinoids are delivered via an occlusive body (i.e., a patch) to alleviate harmful side effects and avoid gastrointestinal (first-pass) metabolism of the drug by the patient. A first aspect of the invention provides a method for relieving symptoms associated with illness or associated with the treatment of illness in a mammalian subject, comprising the steps of selecting at least one cannabinoid from the group consisting of cannabinol, cannabidiol, nabilone, levonantradol, (−)-HU-210, (+)-HU-210, 11-hydroxy-Δ9-THC, Δ8-THC-11-oic acid, CP 55,940, and R(+)-WIN 55,212-2, selecting at least one permeation enhancer from the group consisting of propylene glycol monolaurate, diethylene glycol monoethyl ether, an oleoyl macrogolglyceride, a caprylocaproyl macrogolglyceride, and an oleyl alcohol, and delivering the selected cannabinoid and permeation enhancer transdermally to treat an illness.
Thesis
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The purpose of this research was to develop DNA based techniques for the analysis of botanical forensic evidence. The research included the analysis of Cannabis, the investigation of illegal logging and the analysis of plant material as trace evidence. A set of single nucleotide polymorphisms (SNPs) within the tetrahydrocannabinolic acid synthase gene, corresponding to functional and non-functional forms of the enzyme, were assayed using a mini-sequencing approach (SNaPshot). This enabled the differentiation of Cannabis containing high levels of the psychoactive agent tetrahydrocannabinol (THC) from low THC Cannabis. A multiplex of eleven STR loci was developed and tested across a set of 82 New Zealand Cannabis samples. Typing of New Zealand Cannabis revealed significant departure from Hardy-Weinberg equilibrium, likely due to the effect of clonal reproduction on genotypic diversity in the New Zealand Cannabis population. Unusual triallelic patterns were observed at some loci in a small number of individuals. Sequencing of homozygotes revealed that while most alleles contained the expected repeat region point mutations and insertion/deletion events were also present in some alleles. In one notable example the published repeat sequence did not appear to vary. A set of 13 Inter-simple sequence repeat (ISSR) primers were amplified in the native forest tree Rimu (Dacrydium cupressinum). Although the ISSR primers tested were variable the length of the amplified fragments was considered unsuitable for the analysis of Rimu in the context of illegal logging. A set of primers were designed to target taxon specific fragments of the internal transcribed spacer (ITS) of the nuclear ribosomal DNA (nrDNA). Amplification of taxon specific ITS fragments from experimentally derived mixed samples was successful, however difficulties were encountered with the differentiation of similarly sized amplicons with the microfluidics platform chosen. Of the three aspects of DNA analysis forensic botanical material covered in this thesis the DNA analysis of Cannabis material shows the greatest promise. Although the techniques developed for the analysis of plant fragments from mixed samples and the analysis of Rimu in the context of illegal logging were less successful there is great potential for the use of DNA technologies for the analysis of these evidence types as well.
Article
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Aktuell findet in Deutschland und weiteren westlichen Ländern eine teils sehr emotional geführte Diskussion um eine (weiterführende) Legalisierung von Cannabis als Arzneimittel, aber auch zu Genuss- und Rauschzwecken statt. Von den Gegnern einer Legalisierung wird hierbei oftmals die His­torie der Droge übersehen, die Jahrtausende zurückreicht und sich in verschiedenen Hochkulturen weltweit nachweisen lässt. Ausgeklammert wird ebenso häufig, unter welchen Umständen es überhaupt im 20. Jh. zu einem nahezu globalen Verbot kommen konnte. Die Befürworter hingegen überzeichnen nicht selten den tatsächlich belegten medizinischen Nutzen und verdrängen dabei auch meist die unerwünschten Wirkungen der Droge. Dieser Artikel soll eine sachlichere Diskussion anregen, aber auch auf den noch bestehenden Forschungsbedarf hinweisen.
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o-Quinonemethides generated by condensation of 6-methylhept-5-en-2-ol with various salicylaldehydes in the presence of 30 mol% of fluoroboric acid etherate or triflic acid undergo [4+2]-cycloaddition reaction in situ to afford angularly fused pyranochromene derivatives in good yields and excellent diastereoselectivities.
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Cannabinol, a pharmaceutically interesting component of cannabis, was prepared by a modified Ullmann-Ziegler cross-coupling. Using easily obtainable starting materials, this convergent approach allows facile access to a variety of cannabinol derivatives.
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Objective: To investigate the effects of cannabidiol (CBD) on mitochondrial complex and creatine kinase (CK) activity in the rat brain using spectrophotometry. Method: Male adult Wistar rats were given intraperitoneal injections of vehicle or CBD (15, 30, or 60 mg/kg) in an acute (single dose) or chronic (once daily for 14 consecutive days) regimen. The activities of mitochondrial complexes and CK were measured in the hippocampus, striatum, and prefrontal cortex. Results: Both acute and chronic injection of CBD increased the activity of the mitochondrial complexes (I, II, II-III, and IV) and CK in the rat brain. Conclusions: Considering that metabolism impairment is certainly involved in the pathophysiology of mood disorders, the modulation of energy metabolism (e.g., by increased mitochondrial complex and CK activity) by CBD could be an important mechanism implicated in the action of CBD.
Article
The cannabinoid receptor type 2 (CB2), is a class A GPCR that was cloned in 1993 while looking for an alternate receptor that could explain the pharmacological properties of 9- tetrahydrocannabinol. CB2 was identified among cDNAs based on its similarity in amino-acid sequence to the CB1 receptor and helped provide an explanation for the established effects of cannabinoids on the immune system. In addition to the immune system, CB2 has widespread tissue expression and has been found in brain, PNS and GI tract. Several "mixed" cannabinoid agonists are currently in clinical use primarily for controlling pain and it is believed that selective CB2 agonism may afford a superior analgesic agent devoid of the centrally mediated CB1 effects. Thus, selective CB2 receptor agonists represent high value putative therapeutics for treating pain and other disease states. In this perspective, we seek to provide a concise update of progress in the field.
Article
The CB1 receptor is arguably one of the most abundant GPCRs in the CNS and has long been attractive as a therapeutic target for a wide variety of therapeutic indications including pain, weight gain, emesis and mood disorders. Its cousin, the CB2 receptor, is highly localized in the immune cells regulating immune function and inflammatory pain. Direct acting nonselective agonists, while providing potentially broad therapeutic efficacy, also cause undesirable sedative and hypnotic side effects. New approaches to leverage cannabinoid biology for therapeutic benefit show promise of providing the therapeutic benefits without the buzz.
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A potent, synthetic cannabinoid was radiolabeled and used to characterize and precisely localize cannabinoid receptors in slide-mounted sections of rat brain and pituitary. Assay conditions for 3H-CP55,940 binding in Tris-HCl buffer with 5% BSA were optimized, association and dissociation rate constants determined, and the equilibrium dissociation constant (Kd) calculated (21 nM by liquid scintillation counting, 5.2 nM by quantitative autoradiography). The results of competition studies, using several synthetic cannabinoids, add to prior data showing enantioselectivity of binding and correlation of in vitro potencies with potencies in biological assays of cannabinoid actions. Inhibition of binding by guanine nucleotides was selective and profound: Nonhydrolyzable analogs of GTP and GDP inhibited binding by greater than 90%, and GMP and the nonhydrolyzable ATP analog showed no inhibition. Autoradiography showed great heterogeneity of binding in patterns of labeling that closely conform to cytoarchitectural and functional domains. Very dense 3H-CP55,940 binding is localized to the basal ganglia (lateral caudate-putamen, globus pallidus, entopeduncular nucleus, substantia nigra pars reticulata), cerebellar molecular layer, innermost layers of the olfactory bulb, and portions of the hippocampal formation (CA3 and dentate gyrus molecular layer). Moderately dense binding is found throughout the remaining forebrain. Sparse binding characterizes the brain stem and spinal cord. Densitometry confirmed the quantitative heterogeneity of cannabinoid receptors (10 nM 3H-CP55,940 binding ranged in density from 6.3 pmol/mg protein in the substantia nigra pars reticulata to 0.15 pmol/mg protein in the anterior lobe of the pituitary). The results suggest that the presently characterized cannabinoid receptor mediates physiological and behavioral effects of natural and synthetic cannabinoids, because it is strongly coupled to guanine nucleotide regulatory proteins and is discretely localized to cortical, basal ganglia, and cerebellar structures involved with cognition and movement.
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Cannabis is now emerging from a period of prohibition and being revisited as a potential source of treatments for conditions ill served by synthetic substances. Previous research focussed primarily on effects produced by synthetic cannabinoids such as THC, or cannabis of unknown cannabinoid content. Chemovars of cannabis characterized by high content of specific cannabinoids (primarily, but not only THC and CBD) have been developed. Clinical research using defined extracts from these chemovars is now underway in the UK.Many diseases are multifactorial; a variety of receptors need to be targeted to produce a therapeutic effect. A defined botanical may better achieve this than a single synthetic compound as the components can act synergistically. A new generation of cannabis based medicinal products takes advantage of increasing understanding of the mode of action of cannabinoids, evidence-based research on clinical uses and new technology for realization of products, in anti-diversionary presentations.
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(-)-Δ⁹-Tetrahydrocannabinol ((-)-Δ⁹-THC) is the major active psychotropic component of the marijuana plant, Cannabis sativa. The membrane proteins that have been found to bind this material or its derivatives have been called the cannabinoid receptors. Two GTP-binding protein-coupled cannabinoid receptors have been cloned. CB1 or the neuronal cannabinoid receptor is found mostly in neuronal cells and tissues while CB2 or the peripheral cannabinoid receptor has been detected in spleen and in several cells of the immune system. It has previously been shown that activation of CB1 or CB2 receptors by cannabinoid agonists inhibits adenylyl cyclase activity. Utilizing Chinese hamster ovary cells and COS cells transfected with the cannabinoid receptors we report that(-)-Δ⁹-THC binds to both receptors with similar affinity. However, in contrast to its capacity to serve as an agonist for the CB1 receptor, (-)-Δ⁹-THC was only able to induce a very slight inhibition of adenylyl cyclase at the CB2 receptor. Morever, (-)-Δ⁹-THC antagonizes the agonist-induced inhibition of adenylyl cyclase mediated by CB2. Therefore, we conclude that (-)-Δ⁹-THC constitutes a weak antagonist for the CB2 receptor.
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Two cannabinoid receptors have been identified: CB1, present in the central nervous system (CNS) and to a lesser extent in other tissues, and CB2, present outside the CNS, in peripheral organs. There is evidence for the presence of CB2-like receptors in peripheral nerve terminals. We report now that we have synthesized a CB2-specific agonist, code-named HU-308. This cannabinoid does not bind to CB1\ (Ki>10\ mu M), but does so efficiently to CB2\ (Ki=22.7± 3.9\ nM); it inhibits forskolin-stimulated cyclic AMP production in CB2-transfected cells, but does so much less in CB1-transfected cells. HU-308 shows no activity in mice in a tetrad of behavioral tests, which together have been shown to be specific for tetrahydrocannabinol (THC)-type activity in the CNS mediated by CB1. However, HU-308 reduces blood pressure, blocks defecation, and elicits anti-inflammatory and peripheral analgesic activity. The hypotension, the inhibition of defecation, the anti-inflammatory and peripheral analgesic effects produced by HU-308 are blocked (or partially blocked) by the CB2 antagonist SR-144528, but not by the CB1 antagonist SR-141716A. These results demonstrate the feasibility of discovering novel nonpsychotropic cannabinoids that may lead to new therapies for hypertension, inflammation, and pain.
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In glaucoma, the increased release of glutamate is the major cause of retinal ganglion cell death. Cannabinoids have been demonstrated to protect neuron cultures from glutamate-induced death. In this study, we test the hypothesis that glutamate causes apoptosis of retinal neurons via the excessive formation of peroxynitrite, and that the neuroprotective effect of the psychotropic Delta9-tetrahydroxycannabinol (THC) or nonpsychotropic cannabidiol (CBD) is via the attenuation of this formation. Excitotoxicity of the retina was induced by intravitreal injection of N-methyl-D-aspartate (NMDA) in rats, which also received 4-hydroxy-2,2,6,6-tetramethylpiperidine-n-oxyl (TEMPOL,a superoxide dismutase-mimetic), N-omega-nitro-L-arginine methyl ester (L-NAME, a nitric oxide synthase inhibitor), THC, or CBD. Retinal neuron loss was determined by TDT-mediated dUTP nick-end labeling assay, inner retinal thickness, and quantification of the mRNAs of ganglion cell markers. NMDA induced a dose- and time-dependent accumulation of nitrite/nitrate, lipid peroxidation, and nitrotyrosine (foot print of peroxynitrite), and a dose-dependent apoptosis and loss of inner retinal neurons. Treatment with L-NAME or TEMPOL protected retinal neurons and confirmed the involvement of peroxynitrite in retinal neurotoxicity. The neuroprotection by THC and CBD was because of attenuation of peroxynitrite. The effect of THC was in part mediated by the cannabinoid receptor CB1. These results suggest the potential use of CBD as a novel topical therapy for the treatment of glaucoma.
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A potent, synthetic cannabinoid was radiolabeled and used to characterize and precisely localize cannabinoid receptors in slide-mounted sections of rat brain and pituitary. Assay conditions for 3H-CP55,940 binding in Tris-HCl buffer with 5% BSA were optimized, association and dissociation rate constants determined, and the equilibrium dissociation constant (Kd) calculated (21 nM by liquid scintillation counting, 5.2 nM by quantitative autoradiography). The results of competition studies, using several synthetic cannabinoids, add to prior data showing enantioselectivity of binding and correlation of in vitro potencies with potencies in biological assays of cannabinoid actions. Inhibition of binding by guanine nucleotides was selective and profound: Nonhydrolyzable analogs of GTP and GDP inhibited binding by greater than 90%, and GMP and the nonhydrolyzable ATP analog showed no inhibition. Autoradiography showed great heterogeneity of binding in patterns of labeling that closely conform to cytoarchitectural and functional domains. Very dense 3H-CP55,940 binding is localized to the basal ganglia (lateral caudate-putamen, globus pallidus, entopeduncular nucleus, substantia nigra pars reticulata), cerebellar molecular layer, innermost layers of the olfactory bulb, and portions of the hippocampal formation (CA3 and dentate gyrus molecular layer). Moderately dense binding is found throughout the remaining forebrain. Sparse binding characterizes the brain stem and spinal cord. Densitometry confirmed the quantitative heterogeneity of cannabinoid receptors (10 nM 3H-CP55,940 binding ranged in density from 6.3 pmol/mg protein in the substantia nigra pars reticulata to 0.15 pmol/mg protein in the anterior lobe of the pituitary). The results suggest that the presently characterized cannabinoid receptor mediates physiological and behavioral effects of natural and synthetic cannabinoids, because it is strongly coupled to guanine nucleotide regulatory proteins and is discretely localized to cortical, basal ganglia, and cerebellar structures involved with cognition and movement.
Article
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The neuroprotective actions of cannabidiol and other cannabinoids were examined in rat cortical neuron cultures exposed to toxic levels of the excitatory neurotransmitter glutamate. Glutamate toxicity was reduced by both cannabidiol, a nonpsychoactive constituent of marijuana, and the psychotropic cannabinoid (-)Delta9-tetrahydrocannabinol (THC). Cannabinoids protected equally well against neurotoxicity mediated by N-methyl-D-aspartate receptors, 2-amino-3-(4-butyl-3-hydroxyisoxazol-5-yl)propionic acid receptors, or kainate receptors. N-methyl-D-aspartate receptor-induced toxicity has been shown to be calcium dependent; this study demonstrates that 2-amino-3-(4-butyl-3-hydroxyisoxazol-5-yl)propionic acid/kainate receptor-type neurotoxicity is also calcium-dependent, partly mediated by voltage sensitive calcium channels. The neuroprotection observed with cannabidiol and THC was unaffected by cannabinoid receptor antagonist, indicating it to be cannabinoid receptor independent. Previous studies have shown that glutamate toxicity may be prevented by antioxidants. Cannabidiol, THC and several synthetic cannabinoids all were demonstrated to be antioxidants by cyclic voltametry. Cannabidiol and THC also were shown to prevent hydroperoxide-induced oxidative damage as well as or better than other antioxidants in a chemical (Fenton reaction) system and neuronal cultures. Cannabidiol was more protective against glutamate neurotoxicity than either ascorbate or alpha-tocopherol, indicating it to be a potent antioxidant. These data also suggest that the naturally occurring, nonpsychotropic cannabinoid, cannabidiol, may be a potentially useful therapeutic agent for the treatment of oxidative neurological disorders such as cerebral ischemia.
Article
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Two cannabinoid receptors have been identified: CB(1), present in the central nervous system (CNS) and to a lesser extent in other tissues, and CB(2), present outside the CNS, in peripheral organs. There is evidence for the presence of CB(2)-like receptors in peripheral nerve terminals. We report now that we have synthesized a CB(2)-specific agonist, code-named HU-308. This cannabinoid does not bind to CB(1) (K(i) > 10 microM), but does so efficiently to CB(2) (K(i) = 22.7 +/- 3.9 nM); it inhibits forskolin-stimulated cyclic AMP production in CB(2)-transfected cells, but does so much less in CB(1)-transfected cells. HU-308 shows no activity in mice in a tetrad of behavioral tests, which together have been shown to be specific for tetrahydrocannabinol (THC)-type activity in the CNS mediated by CB(1). However, HU-308 reduces blood pressure, blocks defecation, and elicits anti-inflammatory and peripheral analgesic activity. The hypotension, the inhibition of defecation, the anti-inflammatory and peripheral analgesic effects produced by HU-308 are blocked (or partially blocked) by the CB(2) antagonist SR-144528, but not by the CB(1) antagonist SR-141716A. These results demonstrate the feasibility of discovering novel nonpsychotropic cannabinoids that may lead to new therapies for hypertension, inflammation, and pain.
Article
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Two types of cannabinoid receptor have been discovered so far, CB(1) (2.1: CBD:1:CB1:), cloned in 1990, and CB(2) (2.1:CBD:2:CB2:), cloned in 1993. Distinction between these receptors is based on differences in their predicted amino acid sequence, signaling mechanisms, tissue distribution, and sensitivity to certain potent agonists and antagonists that show marked selectivity for one or the other receptor type. Cannabinoid receptors CB(1) and CB(2) exhibit 48% amino acid sequence identity. Both receptor types are coupled through G proteins to adenylyl cyclase and mitogen-activated protein kinase. CB(1) receptors are also coupled through G proteins to several types of calcium and potassium channels. These receptors exist primarily on central and peripheral neurons, one of their functions being to inhibit neurotransmitter release. Indeed, endogenous CB(1) agonists probably serve as retrograde synaptic messengers. CB(2) receptors are present mainly on immune cells. Such cells also express CB(1) receptors, albeit to a lesser extent, with both receptor types exerting a broad spectrum of immune effects that includes modulation of cytokine release. Of several endogenous agonists for cannabinoid receptors identified thus far, the most notable are arachidonoylethanolamide, 2-arachidonoylglycerol, and 2-arachidonylglyceryl ether. It is unclear whether these eicosanoid molecules are the only, or primary, endogenous agonists. Hence, we consider it premature to rename cannabinoid receptors after an endogenous agonist as is recommended by the International Union of Pharmacology Committee on Receptor Nomenclature and Drug Classification. Although pharmacological evidence for the existence of additional types of cannabinoid receptor is emerging, other kinds of supporting evidence are still lacking.
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Although Delta(9)-tetrahydrocannabinol (THC) produces analgesia, its effects on nociceptive primary afferents are unknown. These neurons participate not only in pain signaling but also in the local response to tissue injury. Here, we show that THC and cannabinol induce a CB(1)/CB(2) cannabinoid receptor-independent release of calcitonin gene-related peptide from capsaicin-sensitive perivascular sensory nerves. Other psychotropic cannabinoids cannot mimic this action. The vanilloid receptor antagonist ruthenium red abolishes the responses to THC and cannabinol. However, the effect of THC on sensory nerves is intact in vanilloid receptor subtype 1 gene knock-out mice. The THC response depends on extracellular calcium but does not involve known voltage-operated calcium channels, glutamate receptors, or protein kinases A and C. These results may indicate the presence of a novel cannabinoid receptor/ion channel in the pain pathway.
Article
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Delta(9)-tetrahydrocannabinol (THC), the main psychoactive component of marijuana has been shown to suppress the immune response. However, the exact mechanism of THC-induced immunosuppression remains unclear. In the current study, we tested the hypothesis that exposure to THC leads to the induction of apoptosis in lymphocyte populations. Splenocytes of C57BL/6 mice cultured in the presence of 10 microM or greater concentrations of THC showed significantly reduced proliferative response to mitogens, including anti-CD3 monoclonal antibodies (mAbs), concanavalin A (Con A), and lipopolysaccharide (LPS) in vitro. Thymocytes and naive and activated splenocytes exposed to 10 microM or 20 microM THC showed significantly increased levels of apoptosis. Treatment with CB2 antagonist inhibited THC-induced apoptosis in thymocytes and activated splenocytes. Administration of 10 mg/kg body weight of THC into C57BL/6 mice led to thymic and splenic atrophy as early as 6 h after treatment. This effect could be partially inhibited by treatment with a caspase inhibitor in vivo. THC exposure led to reductions in the numbers of all subpopulations of splenocytes and thymocytes examined. Functional studies revealed that splenocytes from THC-treated mice had significantly reduced proliferative response to anti-CD3 mAbs, Con A, and LPS in vitro. Finally, thymocytes and splenocytes exposed to THC in vivo exhibited apoptosis upon in vitro culture. Together, these results suggest that in vivo exposure to THC can lead to significant suppression of the immune response by induction of apoptosis.
Chapter
Many of the effects of cannabinoids are mediated by specific cannabinoid receptors, two types of which have so far been identified: CB b cloned in 1990, and CB 2, cloned in 1993 (1,2). Both these receptor types are negatively coupled to adenylate cyclase through Gila proteins and positively coupled to mitogen-activated protein kinase. In addition, CB 1 receptors are positively coupled to inwardly rectifying and A-type potassium channels and negatively coupled to Ntype and P/Q-type calcium channels, again through Gila proteins. Other effector systems for the CB 1 receptor have also been proposed (1,2). CB 1 receptors are present in the central nervous system (CNS) as well as in certain neuronal and nonneuronal peripheral tissues. Their distribution in brain and spinal cord is heterogeneous and unlike that for any other receptor type, with central areas expressing CB I receptors, including some that are implicated in the processing of nociceptive stimuli (1,3). At the neuronal level, some central and peripheral CB 1 receptors are located at nerve terminals where they probably modulate neurotransmitter release when activated (1-3). In contrast, CB 2 receptors are found mainly in cells of the immune system. Another important recent discovery has been that mammalian tissues produce cannabinoid receptor agonists, the most important of which are arachidonoyl ethanolarnide (anandarnide) and 2-arachidonoyl glycerol (1,2). Endogenous cannabinoids and their receptors constitute the endogenous cannabinoid system.
Article
There is now convincing evidence for the existence of at least two types of cannabinoid receptor, CB1 and CB2, both coupled to G proteins. CB1 receptors are present in the central nervous system and in certain peripheral tissues where at least some are located at autonomic nerve terminals. CB2 receptors are found only outside the brain, mainly in cells of the immune system. The existence of endogenous ligands for cannabinoid receptors, both centrally and peripherally, is also generally accepted. These recent discoveries have prompted the development of cannabinoid receptor antagonists and of selective cannabinoid CB1 and CB2 receptor agonists. These compounds are important experimental tools that will help to establish the physiological roles of cannabinoid receptors and their endogenous ligands. The availability of such compounds should also facilitate the discovery of novel therapeutic uses for cannabinoid receptor agonists and antagonists.
Conference Paper
Professor Roger Pertwee will review the state of cannabinoid pharmacology from receptors and ligands to behavioural effects, by describing recently discovered actions of plant-derived cannabinoids that have revealed promising novel clinical uses of one or other of these cannabinoids for the improved treatment of several serious disorders that affect the brain or other parts of the body. Discussion will focus on the strength of the evidence for these potential new areas of cannabinoid therapeutics.
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Cannabinoids, the active components of Cannabis sativa L., act in the body by mimicking endo- genous substances - the endocannabinoids - that activate specific cell surface receptors. Cannabi- noids exert palliative effects in cancer patients. For example, they inhibit chemotherapy-induced nausea and vomiting, stimulate appetite and inhibit pain. In addition, cannabinoids inhibit tumor growth in laboratory animals. They do so by modulating key cell signaling pathways, thereby in- ducing antitumoral actions such as the apoptotic death of tumor cells as well as the inhibition of tumor angiogenesis. Of interest, cannabinoids seem to be selective antitumoral compounds as they can kill tumor cells without significantly affecting the viability of their non-transformed counter- parts. On the basis of these preclinical findings a pilot clinical study of ∆ 9 -tetrahydrocannabinol (THC) in patients with recurrent glioblastoma multiforme has recently been run. The fair safety profile of THC, together with its possible growth-inhibiting action on tumor cells, may set the ba- sis for future trials aimed at evaluating the potential antitumoral activity of cannabinoids.
Article
Endocannabinoids are thought to act as retrograde messengers in the central nervous system. By activating presynaptic cannabinoid CB1 receptors they can reduce neurotransmitter release and modulate synaptic plasticity. To date, anandamide and 2-arachidonoylglycerol (2-AG) are the best studied endocannabinoids. The life span of these lipid molecules in the extracellular space is an important factor in the regulation of their cellular responses. In this review we will discuss the metabolic fate of endocannabinoids, i.e. the mechanisms leading to the termination and/or modification of their actions. It is thought that endocannabinoids can be inactivated via a two-step mechanism. First, endocannabinoids are proposed to be translocated into the cell via selective transporter(s). However, the elusive nature of the putative protein responsible for endocannabinoid uptake has initiated a debate on its existence. Evidence in favor and against will be discussed. Once inside the cell, two major metabolic pathways act upon endocannabinoids: hydrolysis and oxygenation. Hydrolysis of the amide or ester function in anandamide and 2-AG, respectively, terminates their activity on cannabinoid receptors. The proteins responsible for their hydrolysis, fatty acid amide hydrolase and monoacylglycerol lipase, have been cloned and studied in detail. Much less is known about the oxygenation pathways. Lipoxygenase- and cycloxygenase-catalyzed oxygenation of endocannabinoids has been shown to generate a new array of possible biologically active compounds, such as the prostamides and the prostaglandin-glycerols, acting upon novel molecular targets. We will discuss the formation and the possible actions of these novel endocannabinoid derivatives.
Article
There is good evidence that plant-derived and synthetic cannabinoids possess neuroprotective properties. These compounds, as a result of effects upon CB1 cannabinoid receptors, reduce the release of glutamate, and in addition reduce the influx of calcium following NMDA receptor activation. The major obstacle to the therapeutic utilization of such compounds are their psychotropic effects, which are also brought about by actions on CB1 receptors. However, synthesis of the endogenous cannabinoids anandamide and 2-arachidonoylglycerol, which also have neuroprotective properties, are increased under conditions of severe inflammation and ischemia, raising the possibility that compounds that prevent their metabolism may be of therapeutic utility without having the drawback of producing psychotropic effects. In this review, the evidence indicating neuroprotective actions of plant-derived, synthetic and endogenous cannabinoids is presented. In addition, the pharmacological properties of endogenous anandamide-related compounds that are not active upon cannabinoid receptors, but which are also produced during conditions of severe inflammation and ischemia and may contribute to a neuroprotective action are reviewed.
Article
There is now convincing evidence for the existence of at least two types of cannabinoid receptor, CB1 and CB2, both coupled to G proteins. CB1 receptors are present in the central nervous system and in certain peripheral tissues where at least some are located at autonomic nerve terminals. CB2 receptors are found only outside the brain, mainly in cells of the immune system. The existence of endogenous ligands for cannabinoid receptors, both centrally and peripherally, is also generally accepted. These recent discoveries have prompted the development of cannabinoid receptor antagonists and of selective cannabinoid CB1 and CB2 receptor agonists. These compounds are important experimental tools that will help to establish the physiological roles of cannabinoid receptors and their endogenous ligands. The availability of such compounds should also facilitate the discovery of novel therapeutic uses for cannabinoid receptor agonists and antagonists.
Article
There is convincing evidence that mammalian tissues express at least two types of cannabinoid receptor, CB1 and CB2, and that the endogenous cannabinoid, anandamide, and certain other eicosanoid agonists for known cannabinoid receptors can also activate vanilloid (VR1) receptors. Evidence is now also emerging that in addition to these established receptors for cannabinoids, other pharmacological targets for eicosanoid and / or non-eicosanoid cannabinoids are present in neuronal or non-neuronal tissues that include brain, spinal cord, microglial cells, heart, certain arteries, small intestine, vas deferens and peritoneum. Among new receptors to have been proposed for cannabinoids are CB2-like receptors in mouse paw and peritoneum, receptors for abnormal-cannabidiol in microglial cells and in arterial endothelial and non-endothelial cells, Gprotein coupled receptors for R-(+)-WIN55212 and anandamide in brain and spinal cord, receptors for 9- tetrahydrocannabinol and cannabinol on perivascular sensory nerves, 2-adrenoceptor-like (imidazoline) receptors at sympathetic nerve terminals and VR1-like receptors on glutamatergic neurons in hippocampus and dentate gyrus. The presence of novel allosteric sites for cannabinoids on delayed rectifier potassium channels and on 5-HT3, muscarinic M1 and M4, and glutamate GLUA1 and GLUA3 receptors has also been proposed. Current evidence for the existence of these new molecular targets for cannabinoids is summarized in this review. This evidence is largely pharmacological in nature, much of it coming from functional or binding assays with established or novel ligands, sometimes performed using tissues or cell lines that do not express CB1 or CB2 receptors. None of the proposed new cannabinoid receptors have yet been cloned.
Article
The use of cannabinoids for medicinal purposes has attracted wide interest in recent years, especially after the discovery of the endocannabinoid system. However, mechanisms relevant for neuroprotection and recovery of both exogenously applied and endogenous cannabinoids are only partly established. Supported by the observation that cannabinoids are released after neuronal injury and disorder, various CNS applications are currently investigated and both clinical and experimental data are now emerging. The present review describes possible mechanisms of cannabinoid actions relevant for neuroprotection, with a focus on excitotoxic cascades, oxidative stress and inflammation. The complexity of the system is outlined with regards to different cannabinoids, the receptors and cascades they activate, as well as the cell types and experimental approaches utilised to study their action. Furthermore, the unresolved problem of targeting protective pathways, while avoiding psychoactive and detrimental cascades is discussed.
Article
Cannabinoids - the active components of Cannabis sativa and their derivatives - exert palliative effects in cancer patients by preventing nausea, vomiting and pain and by stimulating appetite. In addition, these compounds have been shown to inhibit the growth of tumour cells in culture and animal models by modulating key cell-signalling pathways. Cannabinoids are usually well tolerated, and do not produce the generalized toxic effects of conventional chemotherapies. So, could cannabinoids be used to develop new anticancer therapies?
Article
There are at least two types of cannabinoid receptor, CB1 and CB2, both G protein coupled. CB1 receptors are expressed predominantly at nerve terminals and mediate inhibition of transmitter release. CB2 receptors are found mainly on immune cells, one of their roles being to modulate cytokine release. Endogenous ligands for these receptors (endocannabinoids) also exist. These discoveries have prompted the development of CB1- and CB2-selective agonists and antagonists. The latter include the CB1-selective SR141716A and LY320135 and the CB2-selective SR144528 and AM630, all of which appear to be inverse agonists. Indeed, antagonists without inverse agonist activity have yet to be developed. As most experiments directed at investigating inverse agonism at cannabinoid receptors have been performed with SR141716A, this review focusses on this agent. It presents evidence that the endocannabinoid system is tonically active and that this activity can stem both from ongoing release of endocannabinoids and from the presence of constitutively active CB1 receptors. Thus, SR141716A seems to induce some inverse cannabimimetic effects by opposing responses to endogenously released endocannabinoids and other such effects by decreasing tonic activity induced by constitutively active CB1 receptors. The interaction of SR141716A with constitutively active receptors is discussed in terms of “two-state” and “three-state” models.
Article
Mammalian tissues contain at least two types of cannabinoid receptor, CB₁, found mainly on neurones and CB₂, found mainly in immune cells. Endogenous ligands for these receptors have also been identified. These endocannabinoids and their receptors constitute the endogenous cannabinoid system. Two cannabinoid receptor agonists, Δ⁹-tetrahydrocannabinol and nabilone, are used clinically as anti-emetics or to boost appetite. Additional therapeutic uses of cannabinoids may include the suppression of some multiple sclerosis and spinal injury symptoms, the management of pain, bronchial asthma and glaucoma, and the prevention of neurotoxicity. There are also potential clinical applications for CB₁ receptor antagonists, in the management of acute schizophrenia and cognitive/memory dysfunctions and as appetite suppressants. Future research is likely to be directed at characterizing the endogenous cannabinoid system more completely, at obtaining more conclusive clinical data about cannabinoids with regard to both beneficial and adverse effects, at developing improved cannabinoid formulations and modes of administration for use in the clinic and at devising clinical strategies for separating out the sought-after effects of CB₁ receptor agonists from their psychotropic and other unwanted effects.
Article
Background: Multiple sclerosis is associated with muscle stiffness, spasms, pain, and tremor. Much anecdotal evidence suggests that cannabinoids could help these symptoms. Our aim was to test the notion that cannabinoids have a beneficial effect on spasticity and other symptoms related to multiple sclerosis. Methods: We did a randomised, placebo-controlled trial, to which we enrolled 667 patients with stable multiple sclerosis and muscle spasticity. 630 participants were treated at 33 UK centres with oral cannabis extract (n=211), Δ9-tetrahydrocannabinol (Δ9-THC; n=206), or placebo (n=213). Trial duration was 15 weeks. Our primary outcome measure was change in overall spasticity scores, using the Ashworth scale. Analysis was by intention to treat. Findings: 611 of 630 patients were followed up for the primary endpoint. We noted no treatment effect of cannabinoids on the primary outcome (p=0·40). The estimated difference in mean reduction in total Ashworth score for participants taking cannabis extract compared with placebo was 0·32 (95% CI −1·04 to 1·67), and for those taking Δ9-THC versus placebo it was 0·94 (−0·44 to 2·31). There was evidence of a treatment effect on patient-reported spasticity and pain (p=0·003), with improvement in spasticity reported in 61% (n=121, 95% CI 54·6–68·2), 60% (n=108, 52·5–66·8), and 46% (n=91, 39·0–52·9) of participants on cannabis extract, Δ9-THC, and placebo, respectively. Interpretation: Treatment with cannabinoids did not have a beneficial effect on spasticity when assessed with the Ashworth scale. However, though there was a degree of unmasking among the patients in the active treatment groups, objective improvement in mobility and patients' opinion of an improvement in pain suggest cannabinoids might be clinically useful.
Article
Vasa deferentia taken from mice treated with delta 9-tetrahydrocannabinol (20 mg/kg i.p., once daily for 2 days) showed tolerance to the inhibitory effect of the cannabinoid, R-(+)-arachidonyl-1'-hydroxy-2'-propylamide, on electrically evoked twitches. This treatment did not induce tolerance to the inhibitory effects on the twitch response of morphine or clonidine or of selective mu-, delta- or kappa-opioid receptor agonists. Nor did it affect the contractile potencies of noradrenaline or beta,gamma-methylene-L-ATP. We suggest that cannabinoid tolerance in the vas deferens is attributable neither to downregulation of opioid receptors or alpha 2-adrenoceptors nor to an increased sensitivity of this tissue to its main contractile transmitters noradrenaline and ATP. A concentration of delta 9-tetrahydrocannabinol that inhibits electrically evoked twitches of the vas deferens (100 nM) did not alter the ability of noradrenaline or beta,gamma-methylene-L-ATP to induce contractions suggesting that delta 9-tetrahydrocannabinol inhibits the twitch response by acting prejunctionally.
Article
Mice pretreated intraperitoneally for 2 days with delta‐9‐tetrahydrocannabinol (delta‐9‐THC) at a dose of 20 mg kg ⁻¹ day ⁻¹ and then challenged intravenously with this drug, 24 h after the second pretreatment, showed a 6 fold tolerance to the hypothermic effect of delta‐9‐THC. This pretreatment also induced tolerance to the hypothermic effects of the cannabimimetic agents, CP 55,940 (4.6 fold) and WIN 55,212‐2 (4.9 fold), but not to the hypothermic effect of the putative endogenous cannabinoid, anandamide. Vasa deferentia removed from mice pretreated intraperitoneally with delta‐9‐THC twice at a dose of 20 mg kg ⁻¹ day ⁻¹ were less sensitive to its inhibitory effect on electrically‐evoked contractions than vasa deferentia obtained from control animals. The cannabinoid pretreatment induced a 30 fold parallel rightward shift in the lower part of the concentration‐response curve of delta‐9‐THC and a marked reduction in the maximal inhibitory effect of the drug. It also induced tolerance to the inhibitory effects on the twitch response of CP 55,940 (8.7 fold), WIN 55,212‐2 (9.6 fold) and anandamide (12.3 fold). The results confirm that cannabinoid tolerance can be rapid in onset and support the hypothesis that it is mainly pharmacodynamic in nature. The finding that in vivo pretreatment with delta‐9‐THC can produce tolerance not only to its own inhibitory effect on the vas deferens but also to that of three other cannabimimetic agents, suggests that this tissue would be suitable as an experimental model for investigating the mechanisms responsible for cannabinoid tolerance. Further experiments are required to establish why tolerance to anandamide‐induced hypothermia was not produced by a pretreatment with delta‐9‐THC that did induce tolerance to the hypothermic effects of delta‐9‐THC, CP 55,940 and WIN 55,212‐2 and to the inhibitory effects of delta‐9‐THC, CP 55,940, WIN 55,212‐2 and anandamide on the twitch response of the vas deferens.
Article
(-)-Delta9-Tetrahydrocannabinol ((-)-Delta9-THC) is the major active psychotropic component of the marijuana plant, Cannabis sativa. The membrane proteins that have been found to bind this material or its derivatives have been called the cannabinoid receptors. Two GTP-binding protein-coupled cannabinoid receptors have been cloned. CB1 or the neuronal cannabinoid receptor is found mostly in neuronal cells and tissues while CB2 or the peripheral cannabinoid receptor has been detected in spleen and in several cells of the immune system. It has previously been shown that activation of CB1 or CB2 receptors by cannabinoid agonists inhibits adenylyl cyclase activity. Utilizing Chinese hamster ovary cells and COS cells transfected with the cannabinoid receptors we report that (-)-Delta9-THC binds to both receptors with similar affinity. However, in contrast to its capacity to serve as an agonist for the CB1 receptor, (-)-Delta9-THC was only able to induce a very slight inhibition of adenylyl cyclase at the CB2 receptor. Morever, (-)-Delta9-THC antagonizes the agonist-induced inhibition of adenylyl cyclase mediated by CB2. Therefore, we conclude that (-)-Delta9-THC constitutes a weak antagonist for the CB2 receptor.
Article
Anandamide has been identified in porcine brain as an endogenous cannabinoid receptor ligand and is believed to be a counterpart to the psychoactive component of marijuana, delta 9-tetrahydrocannabinol (delta 9-THC). Here we report that anandamide directly inhibits (IC50, 2.7 muM) Shaker-related Kv1.2 K+ channels that are found ubiquitously in the mammalian brain. Delta 9-THC also inhibited Kv1.2 channels with comparable potency (IC50, 2.4 muM), as did several N-acyl-ethanolamides with cannabinoid receptor binding activity. Potassium current inhibition occurred through a pertussis toxin-insensitive mechanism and was not prevented by the cannabinoid receptor antagonist SR141716A. Utilizing excised patches of Kv1.2 channel-rich membrane as a rapid and sensitive bioassay, we found that phospholipase D stimulated the release of an endogenous anandamide-like K+ channel blocker from rat brain slices. Structure-activity studies were consistent with the possibility that the released blocker was either anandamide or another N-acyl-ethanolamide.
Article
There are at least two types of cannabinoid receptors, CB1 and CB2, both coupled to G-proteins. CB1 receptors are present in the central nervous system and CB1 and CB2 receptors in certain peripheral tissues. The existence of endogenous cannabinoid receptor agonists has also been demonstrated. These discoveries have led to the development of selective cannabinoid CB1 and CB2 receptor ligands. This review focuses on the classification, binding properties, effector systems and distribution of cannabinoid receptors. It also describes the various cannabinoid receptor agonists and antagonists now available and considers the main in vivo and in vitro bioassay methods that are generally used.
Article
Tolerance develops rapidly to cannabis, cannabinoids, and related drugs acting at the CB1 cannabinoid receptor. However, little is known about what happens to the receptor as tolerance is developing. In this study, we have found that CB1 receptors are rapidly internalized following agonist binding and receptor activation. Efficacious cannabinoid agonists (WIN 55,212-2, CP 55,940, and HU 210) caused rapid internalization. Methanandamide (an analogue of an endogenous cannabinoid, anandamide) was less effective, causing internalization only at high concentration, whereas delta9-tetrahydrocannabinol caused little internalization, even at 3 microM. CB1 internalized via clathrin-coated pits as sequestration was inhibited by hypertonic sucrose. Internalization did not require activated G protein alpha(i), alpha(o), or alpha(s) subunits. A region of the extreme carboxy terminus of the receptor was necessary for internalization, as a mutant CB1 receptor lacking the last 14 residues did not internalize, whereas a mutant lacking the last 10 residues did. Steps involved in the recycling of sequestered receptor were also investigated. Recovery of CB1 to the cell surface after short (20 min) but not long (90 min) agonist treatment was independent of new protein synthesis. Recycling also required endosomal acidification and dephosphorylation. These results show that CB1 receptor trafficking is dynamically regulated by cannabimimetic drugs.