Cannabinoids and endocannabinoids as therapeutics for nervous system disorders:preclinical models and clinical studies

Cannabinoids are lipophilic substances derived from Cannabis sativa that can exert a variety of effects in the human body.They have been studied in cellular and animal models as well as in human clinical trials for their therapeutic benefits in several human diseases.Some of these include central nervous system(CNS)diseases and dysfunctions such as forms of epilepsy,multiple sclerosis,Parkinson’s disease,pain and neuropsychiatric disorders.In addition,the endogenously produced cannabinoid lipids,endocannabinoids,are critical for normal CNS function,and if controlled or modified,may represent an additional therapeutic avenue for CNS diseases.This review discusses in vitro cellular,ex vivo tissue and in vivo animal model studies on cannabinoids and their utility as therapeutics in multiple CNS pathologies.In addition,the review provides an overview on the use of cannabinoids in human clinical trials for a variety of CNS diseases.Cannabinoids and endocannabinoids hold promise for use as disease modifiers and therapeutic agents for the prevention or treatment of neurodegenerative diseases and neurological disorders.

Dispersive liquid-liquid microextraction,an effective tool for the determination of synthetic cannabinoids in oral fluid by liquid chromatography-tandem mass spectrometry

In the present work,dispersive liquid-liquid microextraction(DLLME)was used to extract six synthetic cannabinoids(JWH-018,JWH-019,JWH-073,JWH-200,or WIN 55,225,JWH-250,and AM-694)from oral fluids.A rapid baseline separation of the analytes was achieved on a bidentate octadecyl silica hydride phase(Cogent Bidentate C18;4.6 mm×250 mm,4μm)maintained at 37℃,by eluting in isocratic conditions(water:acetonitrile(25:75,V/V)).Detection was performed using positive electrospray ionization-tandem mass spectrometry.The parameters affecting DLLME(pH and ionic strength of the aqueous phase,type and volume of the extractant and dispersive solvent,vortex and centrifugation time)were optimized for maximizing yields.In particular,using 0.5 mL of oral fluid,acetonitrile(1 mL),was identified as the best option,both as a solvent to precipitate proteins and as a dispersing solvent in the DLLME procedure.To select an extraction solvent,a low transition temperature mixture(LTTM;composed of sesamol and chlorine chloride with a molar ratio of 1:3)and dichloromethane were compared;the latter(100μL)was proved to be a better extractant,with recoveries ranging from 73%to 101%by vortexing for 2 min.The method was validated according to the guidelines of Food and Drug Administration bioanalytical methods:intra-day and inter-day precisions ranged between 4%and 18%depending on the spike level and analyte;limits of detection spanned from 2 to 18 ng/mL;matrixmatched calibration curves were characterized by determination coefficients greater than 0.9914.Finally,the extraction procedure was compared with previous methods and with innovative techniques,presenting superior reliability,rapidity,simplicity,inexpensiveness,and efficiency.

Bilateral renal cortical necrosis associated with smoking synthetic cannabinoids

Synthetic cannabinoids have become a common drug of abuse in recent years and their toxicities have come to light as well. They are known to be notorious for the kidneys, with acute tubular necrosis, acute interstitial nephritis and rhabdomyolysis induced renal injury being the frequent nephrotoxic outcomes in users. We report a case of bilateral renal cortical necrosis, leading to irreversible renal damage and lifelong dialysis dependency.

Synthetic cannabinoids 2015: An update for pediatricians in clinical practice

Synthetic cannabinoids are a group of substances in the world of designer drugs that have become increasingly popular over the past few years. Synthetic cannabinoids are a chemically diverse group of compounds functionally similar to THC. Since first appearing on the world market a few years ago these compounds have evolved rapidly. Newer more potent analogues have been developed. Identifying youth who abuse these substances can be difficult. Newer forms of consumption have also evolved. These products are now manufactured in products that look like natural cannabis resin and in liquid cartridges used in electronic cigarettes. Synthetic cannabinoids appear to be associated with potentially dangerous health effects that are more severe than that of marijuana. Some synthetic cannabinoid compounds have been associated with serious physical consequences, such as, seizures, myocardial infarction and renal damage. In addition, psychoactive effects, such as aggression, confusion, anxiety and psychosis have also been reported. The diagnosis remains primarily clinical with toxicological confirmation difficult due to manufacturers constantly developing new analogues to avoid detection. Pediatricians are urged to familiarize themselves with these drugs and the typical presentations of patients who use them.

Cannabinoids Production by Hairy Root Cultures of <i>Cannabis sativa</i>L.

Tetrahydrocannabinol (THC) derivatives are used clinically as analgesic, anti-inflammatory, appetite stimulant, anti-emetic and anti-tumor cannabinoids. THC and its related compounds are at present obtained by extraction from intact Cannabis plants or chemical synthesis, but plant cell cultures may be an alternative source of production. In the present study, hairy root cultures of C. sativa (Cannabaceae) were induced by incubation of aseptically grown callus culture with solid B5 medium supplemented with 4 mg/l naphthaleneacetic acid in darkness at 25°C. Hairy root growth profiles in shake flask, increased periodically during 35 days of growth cycle. The cannabinoid contents produced in minor levels and remained below 2.0 μg/g dry weight. The contents of can-nabinoid were analyzed by liquid chromatography and confirmed by mass spectrometry.

In vitro Activity of Novel Cannabinoids Derived from Tetrahydrocan-nabinolic Acid on Various Human Tumor Cell Lines

The in vitro study of tetracannabinolic acid(THCA)derivatives ALAM027 and ALAM108 was carried out on the following human tumor cells:T47D(breast,ductal carcinoma),PC-3(prostate,adenocarcinoma),HT 29(colorectal carcinoma),Caco-2(colon,adenocarcinoma),A549(lung,carcinoma),U87MG(human glioblastoma)and U266B1(multiple myeloma).The in vitro effects of THCA derivatives ALAM027 and ALAM108 on cell growth inhibition and IC50 values were measured using the CellTiter Glo assay.The ALAM027 compound showed good growth inhibition in all cell lines tested with the exception of U87MG cells.The ALAM108 compound also suppressed the growth of U87 MG cells but had little effect on T47D tumor cells.In vitro studies of THCA derivatives ALAM027 and ALAM108 showed antitumor activity in all cell lines tested.The difference in the activity of these compounds in relation to the T47D and U87MG tumor cells may be indicative of different functional mechanisms.

Can cannabinoids be a potential therapeutic tool in amyotrophic lateral sclerosis？

Amyotrophic lateral sclerosis（ALS） is the most common degenerative disease of the motor neuron system. Over the last years, a growing interest was aimed to discovery new innovative and safer therapeutic approaches in the ALS treatment. In this context, the bioactive compounds of Cannabis sativa have shown antioxidant, anti-inflammatory and neuroprotective effects in preclinical models of central nervous system disease. However, most of the studies proving the ability of cannabinoids in delay disease progression and prolong survival in ALS were performed in animal model, whereas the few clinical trials that investigated cannabinoids-based medicines were focused only on the alleviation of ALS-related symptoms, not on the control of disease progression. The aim of this report was to provide a short but important overview of evidences that are useful to better characterize the efficacy as well as the molecular pathways modulated by cannabinoids.

Role of cannabinoids and the endocannabinoid system in modulation of diabetic cardiomyopathy

Diabetic complications,chiefly seen in long-term situations,are persistently deleterious to a large extent,requiring multi-factorial risk reduction strategies beyond glycemic control.Diabetic cardiomyopathy is one of the most common deleterious diabetic complications,being the leading cause of mortality among diabetic patients.The mechanisms of diabetic cardiomyopathy are multi-factorial,involving increased oxidative stress,accumulation of advanced glycation end products(AGEs),activation of various pro-inflammatory and cell death signaling pathways,and changes in the composition of extracellular matrix with enhanced cardiac fibrosis.The novel lipid signaling system,the endocannabinoid system,has been implicated in the pathogenesis of diabetes and its complications through its two main receptors:Cannabinoid receptor type 1 and cannabinoid receptor type 2,alongside other components.However,the role of the endocannabinoid system in diabetic cardiomyopathy has not been fully investigated.This review aims to elucidate the possible mechanisms through which cannabinoids and the endocannabinoid system could interact with the pathogenesis and the development of diabetic cardiomyopathy.These mechanisms include oxidative/nitrative stress,inflammation,accumulation of AGEs,cardiac remodeling,and autophagy.A better understanding of the role of cannabinoids and the endocannabinoid system in diabetic cardiomyopathy may provide novel strategies to manipulate such a serious diabetic complication.

Endocannabinoids Anandamide and Its Cannabinoid Receptors in Liver Fibrosis after Murine Schistosomiasis

This study examined endogenous carmabinoid （ECB）-anandamide （AEA） and its cannabinoid receptors （CBR） in mice liver with the development ofschistosomajaponicum. Mice were infected with schistosoma by means of pasting the cercaria onto their abdomens. Liver fibrosis was pathologically confirmed nine weeks after the infection. High performance liquid chromatography （HPLC） was employed to determine the concentration of AEA in the plasma of mice. Immunofluo-rescence was used to detect the expression of CBR1 and CBR2 in liver tissue. Morphological examination showed typical pathological changes, with worm tubercles of schistosoma deposited in the liver tissue, fibrosis around the worm tubercles and infiltration or soakage of inflammatory cells. Also, CBR1 and CBR2 were present in hepatocytes and hepatic sinusoids of the two groups, but they were obviously enhanced in the schistosoma-infected mice. However, the average optical density of CBR1 in the negative control and fibrosis group was 13.28±7.32 and 30.55±7.78, and CBR2 were 28.13±6.42 and 52.29±4.24 （P〈0.05）. The levels of AEA in the fibrosis group were significantly increased as compared with those of the control group. The concentrations of AEA were （0.37±0.07） and （5.67±1.34） ng/mL （P〈0.05）. It is concluded that the expression of endocannabinoids AEA and its cannabinoid receptor CBR were significantly increased in schistosoma-infected mice. Endogenous endocannabinoids may be involved in the development of schistosoma-induced liver fibrosis.

Endogenous cannabinoids may regulate chronic inflammation in aspirin-exacerbated respiratory disease

Aspirin-exacerbated respiratory disease(AERD)is characterized by the triad of chronic rhinosinusitis with nasal polyposis,adult-onset asthma and non-IgE mediated reactions to aspirin and other cyclooxygenase-1(COX-1)inhibitors.Patients with AERD are dependent on COX-1 activity to maintain production of prostaglandin(PG)species,such as PGE2,which maintain physiologic levels of inflammation and limit the production of pro-inflammatory cysteinyl leukotrienes.The endogenous cannabinoid system is a family of immunomodulatory lipids and their innate g-protein coupled receptors that are closely related to arachidonic acid and may modulate inflammation via several pathways,including the direct production of metabolically active prostaglandin glycerol-esters.A recent pilot study has identified the significant up-regulation of the peripherally expressed,type-2 cannabinoid receptor(CB2)in AERD nasal polyps versus control tissues from patients with either allergic fungal rhinosinusitis or no history of chronic sinonasal inflammation.These early findings suggest the involvement of increased endogenous cannabinoid activity in prostaglandin deficient states such as AERD.Future study is needed to explore the significance of these findings,with specific investigation of the impact of CB2 activation on markers of airway inflammation,as well as the potential to measure CB2 expression as a screening biomarker for the evaluation of unrecognized disease.

Handheld Raman Spectrometer for the Rapid Determination of Synthetic Cannabinoids

A handheld laser Raman spectrometer was developed to detect 11 synthetic cannabinoid standards.The spectrometer can be connected with a mobile phone by Bluetooth or a computer by a data cable.The Raman spectra of the synthetic cannabinoids AM‑694,JWH‑019,and nine others were accurately determined by software analysis of characteristic Raman peaks.This technique is highly efficient,accurate,and simple,and it is a valid procedure for rapid on‑site investigations of cannabinoids.

Coffee, Traditional Chinese Medicine and cannabinoids as potential tools for prevention and treatment of hepatocellular carcinoma

In the last decade,the incidence of hepatocellular carcinoma(HCC)is growing in both Europe and United States.Conventional therapies such as liver resection,transplantation,ablation,chemoembolization and sorafenib are not enough to avoid a significant mortality.Many studies suggested the positive effect of caffeine for prevention of HCC.Nevertheless,the amount of therapeutic caffeine and the high-dose safety are unknown.Many authors proposed Traditional Chinese Medicine as preventive and/or curative approach.Although it reveals limits such as the uncertain safety profile and the lack of evidence about a unique product,it shows interesting results in terms of survival and quality of life if given in combination with standard loco-regional therapy.Among the future promises,cannabinoids show interesting background mechanisms of blocking cell proliferation and neoangiogenesis.It is conceivable that in the next years,some natural products may have a role in improving the standards of care of HCC.

Modified Atmospheric Packaging and Its Effect on Postharvest Cannabis Quality

Cannabis sativa L. is used as fiber, food, and medicine in several countries. Though it is illegal for recreational use in most of the world, there are some countries that have legalized production and sale. There is a lot of research on production of cannabis, but less so on storage technologies. Cannabis contains several high value compounds, such as cannabinoids and terpenoids, that are susceptible to degradation via light, temperature, and oxygen. Several studies have explored temperature and light, and industry has adjusted accordingly. However, less is known about oxygen-induced degradation. Biochemical studies have demonstrated oxidative degradation of high value compounds, and many producers use some form of modified atmospheric packaging (MAP) for storage. However, the efficacy of MAP is unclear. The objective of this paper is to review our current understanding of MAP in postharvest cannabis storage and identify avenues where additional research is needed.

Life-threatening constrictive pericarditis accompanied by synthetic cannabinoid use:A case report and literature review

Rationale:Synthetic cannabinoids are increasingly used as recreational drugs and have been associated with adverse cardiovascular effects.However,reports of synthetic cannabinoids accompanied by constrictive pericarditis are limited.Patient’s concern:A 28-year-old male with a history of synthetic cannabinoid(Bonzai)abuse presented with chest discomfort,dyspnea,and lower extremity edema.Investigations revealed reduced left ventricular ejection fraction,elevated inflammatory markers,low electrocardiogram voltages,and atrial fibrillation.Diagnosis:Chest spiral computerized tomography scan and chest X-ray demonstrated pericardial calcification.Cardiac magnetic resonance imaging and right heart catheterization were done to confirm the possibility of constrictive pericarditis.Based on the patient’s addiction history and exclusion of rheumatologic and infectious causes,it was supposed that constrictive pericarditis and cardiomyopathy may be accompanied by synthetic cannabinoid use.Interventions:The patient received standard medical therapy,including loop diuretics for cardiomyopathy and constrictive pericarditis.Catheter ablation was recommended for his rhythm control,and he was planned for close monitoring of clinical and echocardiographic response and evaluation of the need for surgical pericardiectomy in the future.Outcomes:After 6 months follow-up,echocardiographic exam revealed no significant improvement in ventricular function.However,due to the high surgical risk,the patient’s poor compliance,and the continuation of drug abuse,he was not a good candidate for surgery according to our heart team’s decision.Lessons:Synthetic cannabinoids can trigger constrictive pericarditis,and clinicians should consider them when evaluating patients with compatible symptoms and exposure history.Further research on the cardiovascular effects of synthetic cannabinoids is needed and public education on potential harms is warranted.

Cannabinoids:Challenges,opportunities and current techniques towards its extraction and purification for edibles

Due to the multiple health benefits associated with the consumption of cannabinoids,there is a current trend on continuing to explore their biological properties.Cannabidiol(CBD)is likely to be the most investigated endocannabinoid from the cannabis plant.Unfortunately,the purification of CBD from its natural source and psychotropic analogue(tetrahydrocannabinol,THC)is timely challenging for the research community based on the similar physiochemical properties of both molecules.This latter fact becomes relevant for the final application of CBD in edibles as a bioactive compound.Therefore,this paper comprehensively reviews the latest development works(over the last 2-3 years)aiming at the extraction of cannabinoids and purification of CBD using traditional,emerging,and synergistic extraction techniques and strategies.Particular emphasis and discussion have been focused on the innovative extraction techniques and the relevant outcomes with detailed strategies to meet such high extraction rates.Biological properties(at lab-scale and clinical case studies),current practices,progress in legislation and commercialization of CBD are also overviewed.Finally,according to the findings of this review,the future trends and research gaps are also given.

利莫那班对肝纤维化C57小鼠肝组织大麻素受体1及α-SMA表达的影响

目的研究大麻素受体1(CB1)拮抗剂利莫那班对肝纤维化模型C57小鼠肝组织中CB1、α-平滑肌肌动蛋白(α-SMA)表达的影响,及其抗肝纤维化的作用机制。方法 30只C57小鼠随机分为3组,分别为正常对照组、模型对照组及利莫那班组,每组10只。采用四氯化碳腹腔注射诱导形成小鼠肝纤维化模型。造模2周后于继续造模同时,正常对照组和模型对照组每天生理盐水灌胃,利莫那班组用利莫那班灌胃。第8周造模结束时处死小鼠,留取肝脏组织标本,分别进行HE和Masson三色染色,应用免疫组织化学方法检测肝组织中CB1和α-SMA的表达,并进行肝组织纤维化评分(S评分)。结果模型对照组和利莫那班组肝组织S评分、CB1和α-SMA阳性表达量均高于正常对照组(均P<0.05),利莫那班组均低于模型对照组(均P<0.05);正常对照组、模型对照组和利莫那班组CB1评分、α-SMA评分与S评分相互之间均呈正相关(均P<0.05)。结论肝组织CB1的激活可促进肝纤维化的形成,CB1拮抗剂利莫那班通过抑制CB1表达,进而抑制肝星状细胞的增殖和激活,从而起到抗肝纤维化的作用。

Neuronal and peripheral damages induced by synthetic psychoactive substances: an update of recent findings from human and animal studies

Preclinical and clinical studies indicate that synthetic psychoactive substances,in addition to having abuse potential,may elicit toxic effects of varying severity at the peripheral and central levels.Nowadays,toxicity induced by synthetic psychoactive substances poses a serious harm for health,since recreational use of these substances is on the rise among young and adult people.The present review summarizes recent findings on the peripheral and central toxicity elicited by “old” and “new” synthetic psychoactive substances in humans and experimental animals,focusing on amphetamine derivatives,hallucinogen and dissociative drugs and synthetic cannabinoids.

Cannabis Responsive Head Injury Induced Mutiple Disabilities: A Case Report

In recent years cannabinoids and their derivatives have drawn renewed attention because of their diverse pharmacologic activities. We report here one such case, where all types of medical & psychiatric treatment failed to improve the symptoms;however cannabis use was able to bring back this patient to normal productive & meaningful life. The patient was a 47 year old left handed Caucasian had minor subdural hematoma at the posterior vertex and a minor focal subarachnoid haemorrhage following a physical assault. His impairments included cognitive slowing with decreased short term memory, organized skill & language deficit. His physical disabilities included spastic gait (hemiplegic), VII nerve palsy, mild cerebellar dysfunction, blurred vision and easy fatigue. He was unable to return to work or drive. In addition to cognitive deficit, right hemi paresis & dizziness;he had symptoms of anxiety & depression. Cannabis improved his gait and brought back lots of his memory. Within 6 months all his symptoms abated with use of cannabis and he started to look at cannabis as a real medicine. Slowly he also had improvement in cognitive functions, memory, vocabulary and his gait became increasingly better day by day with continued use of cannabis.

Case of cannabinoid hyperemesis syndrome with long-term follow-up

Long-term cannabis use may be associated with attacks of severe nausea and vomiting, and a characteristic learned behavior of compulsive hot bathing, termed cannabinoid hyperemesis syndrome(CHS). Long-term follow-up and prognosis of CHS have not been reported previously. A 44-year-old Caucasian man with a long history of addiction to marijuana presented with chronic abdominal pain complicated by attacks of uncontrollable vomiting for 16 years. He had a compulsion to take scalding hot showers, as many as 15 times a day, to relieve his symptoms. All previous therapies had been ineffective. However, abstinence from marijuana led to rapid and complete resolution of all symptoms and his compulsive hot showering behavior. He has been followed for nine years, and is still doing well without recurrence of symptoms. Physicians should have a high index of suspicion for this under-recognized condition, as excellent long-term prognosis of CHS can be achieved when abstinence is maintained.

Effects of combined cannabidiol(CBD)and hops(Humulus lupulus)terpene extract treatment on RAW 264.7 macrophage viability and inflammatory markers

This study investigates the potential of cannabidiol(CBD),one major cannabinoid of the plant Cannabis sativa,alone and in combination with a terpene-enriched extract from Humulus lupulus(“Hops 1”),on the LPS-response of RAW 264.7 macrophages as an established in vitro model of inflammation.With the present study,we could support earlier findings of the anti-inflammatory potential of CBD,which showed a dose-dependent[0-5μM]reduction in nitric oxide and tumor necrosis factor-alpha(TNF-α)released by LPS-stimulated RAW 264.7 macrophages.Moreover,we observed an additive anti-inflammatory effect after combined CBD[5μM]and hops extract[40μg/mL]treatment.The combination of CBD and Hops 1 showed effects in LPS-stimulated RAW 264.7 cells superior to the single sub-stance treatments and akin to the control hydrocortisone.Furthermore,cellular CBD uptake increased dose-depend-ently in the presence of terpenes from Hops 1 extract.The anti-inflammatory effect of CBD and its cellular uptake positively correlated with terpene concentration,as indicated by comparison with a hemp extract containing both CBD and terpenes.These findings may contribute to the postulations for the so-called“entourage effect”between cannabinoids and terpenes and support the potential of CBD combined with phytomolecules from a non-cannabi-noid source,such as hops,for the treatment of inflammatory diseases.