The non-euphoric phytocannabinoid cannabidivarin counteracts intestinal inflammation in mice and cytokine expression in biopsies from UC pediatric patients

doi:10.1016/j.phrs.2019.104464

Pharmacological Research

Volume 149, November 2019, 104464

https://doi.org/10.1016/j.phrs.2019.104464 Get rights and content

Abstract

Patients with ulcerative colitis (UC) using marijuana have been reported to experience symptomatic benefit. Cannabidivarin (CBDV) is a safe non-psychoactive phytocannabinoid able to activate and desensitize TRPA1, a member of the TRP channels superfamily, which plays a pivotal role in intestinal inflammation. Here, we have investigated the potential intestinal anti-inflammatory effect of CBDV in mice and in biopsies from pediatric patients with active UC. Colonic inflammation was induced in mice by dinitrobenzenesulfonic acid (DNBS). The effect of orally administered CBDV on macroscopic and microscopic damage, inflammatory parameters (i.e. myeloperoxidase activity, intestinal permeability and cytokine production) and faecal microbiota composition, was evaluated 3 days after DNBS administration. TRPA1 expression was studied by RT-PCR in inflamed colons of mice as well as in mucosal colonic biopsies of children with active UC, whose response to incubation with CBDV was also investigated. CBDV attenuates, in a TRPA1-antagonist sensitive manner, DNBS-induced signs of inflammation including neutrophil infiltration, intestinal permeability, and cytokine (i.e. IL-1β, IL-6 and the chemokine MCP-1) production. CBDV also alters the dysregulation of gut microbiota associated to colitis. Finally, CBDV lessens cytokine expression in colonic biopsies from pediatric patients with ulcerative colitis, a condition in which TRPA1 was up-regulated. Our preclinical study shows that CBDV exerts intestinal anti-inflammatory effects in mice via TRPA1, and in children with active UC. Since CBDV has a favorable safety profile in humans, it may be considered for possible clinical trials in patients with UC.

Graphical abstract

Introduction

Inflammatory bowel diseases (IBDs), including ulcerative colitis (UC) and Crohn’s disease (CD), are relapsing and lifelong disorders characterized by chronic inflammation of the gastrointestinal tract. IBDs represent a global health burden that affects millions of people with rising incidence and prevalence worldwide [1,2]. IBD results from a complex interaction between environmental, genetic and epigenetic risk factors that cause an inappropriate mucosal immune response leading to intestinal inflammation. Additionally, recent evidence suggests that gut microbiota dysbiosis is strictly linked to initiation and progression of IBD, although it is still unclear whether this is a primary or secondary occurrence [3].

The plant Cannabis sativa contains over 100 terpenophenolic constituents, known as phytocannabinoids [4,5]. A number of surveys and small clinical trials suggest that patients with IBD use cannabis preparations to alleviate symptoms such as diarrhea, abdominal pain, and loss of appetite [[6], [7], [8], [9], [10], [11]]. Experimentally, single isolated cannabinoids such as delta-9-tetrahydrocannabinol [12], cannabidiol [[12], [13], [14], [15]], cannabigerol [16], and cannabichromene [17] have been shown to attenuate intestinal inflammation.

Cannabidivarin (CBDV) is a non-psychoactive phytocannabionid that was identified by Vollner and coworkers in 1969, although little information on its pharmacology and mode of action has been reported in the subsequent forty years [18]. More recently, pharmacodynamic studies have shown that CBDV shows very little affinity for cannabinoid receptors, but is a potent agonist of the transient receptor potential (TRP) ankyrin type-1 (TRPA1) [19]. TRPA1, a cation-permeable channel, is a member of the TRP superfamily with a pivotal role in intestinal inflammation [20]. Specifically, TRPA1 is expressed in sensory fibers innervating the gastrointestinal tract, mediates gastrointestinal hypersensitivity to mechanical stimuli, and modulates the release of pro-inflammatory peptides [21]. Currently, CBDV is under clinical investigation for epilepsy (NCT02369471) and Autism Spectrum Disorder in children (NCT03202303).

Here, we report that oral CBDV exerts, via TRPA1, anti-inflammatory effects and changes the composition of gut microbiota in the murine model of colitis induced by DNBS. Additionally, CBDV reduces colonic inflammation in biopsies from pediatric patients with IBD, a condition in which we found an unprecedented example of TRPA1 upregulation.

Section snippets

Drugs and reagents

2,4-dinitrobenzenesulfonic acid (DNBS) and myeloperoxidase (MPO) from human leucocytes, were purchased from Sigma Aldrich S.r.l. (Milan, Italy). CBDV [purity by high-performance liquid chromatography (HPLC), 99.0%], was kindly supplied by GW Research Ltd (Cambridge, UK). CBDV was dissolved in ethanol/Tween20/saline (1:1:8) for intraperitoneal (i.p.) injection (60 μl/mouse) and in carboxymethylcellulose (CMC 1%, 150 μl/mouse) for oral gavage administration. The vehicles had no significant

Preventive or curative CBDV (either oral or intraperitoneal) reduces colon weight/colon length ratio in experimental acute colitis

Intracolonic DNBS administration caused a number of signs of inflammation and disease, including increased colon weight/colon length ratio, neutrophil infiltration (as revealed by MPO activity), intestinal permeability, histological damage, up-regulation of pro-inflammatory cytokines and down-regulation of the anti-inflammatory cytokine IL-10 (Fig. 1, Fig. 2, Fig. 3). Intraperitoneal or oral CBDV (0.3–10 mg/kg), given before (preventive protocol, Fig. 1A,B and F) or after (curative protocol,

Discussion

Despite Cannabis treatment being used in the USA by IBD patients to reduce their IBD symptoms [33,34], our study represents the first evidence of CBDV intestinal anti-inflammatory effects. CBDV is the propyl analogue of cannabidiol, the main non-psychoactive cannabinoid isolated from Cannabis sativa [35]. Recently, CBDV has emerged among cannabinoids for its possible clinical use as an antiepileptic drug. In 2015, a Phase 2, double blind, randomized, placebo-controlled study has shown that CBDV

Contributors

E.P. was responsible for acquisition, analysis and interpretation of data; conception and design; and redaction of the manuscript. B.R., F.I., O.A.P., M.D.A., T.V., S.P., L.C., M.L., G.S., F.L., P.O. and R.C. carried out acquisition, analysis and interpretation of data. A.A.I. and V.D. were responsible for conception and design, analysis and interpretation of data, and critical reading of the manuscript. F.B. performed conception and design, analysis and interpretation of data, and redaction of

Funding

This work was supported by GW Research Ltd., UK.

Patient consent

Obtained.

Declaration of Competing Interest

AI, VD and FAI receive research grants from GW Research Ltd., UK.

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¹: Endocannabinoid Research Group.

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The non-euphoric phytocannabinoid cannabidivarin counteracts intestinal inflammation in mice and cytokine expression in biopsies from UC pediatric patients

Abstract

Graphical abstract

Introduction

Section snippets

Drugs and reagents

Preventive or curative CBDV (either oral or intraperitoneal) reduces colon weight/colon length ratio in experimental acute colitis

Discussion

Contributors

Funding

Patient consent

Declaration of Competing Interest

Biochem. Pharmacol.

Trends Pharmacol. Sci.

Gastroenterology

Dig. Liver Dis.

Gastroenterology

Eur. J. Pharmacol.

Drug Alcohol Depend.

Curr. Opin. Microbiol.

Mucosal Immunol.

Eur. J. Pharmacol.

Gastroenterology

Epidemiology and risk factors for IBD, Nature reviews

Gastroenterol. Hepatol. (N Y)

Understanding and preventing the global increase of inflammatory bowel disease

Gastroenterology

Environmental triggers in IBD: a review of progress and evidence, Nature reviews

Gastroenterol. Hepatol. (N Y)

Potential cannabis synergy and phytocannabinoid-terpenoid entourage effects

Br. J. Pharmacol.

Cannabimimetic phytochemicals in the diet - an evolutionary link to food selection and metabolic stress adaptation?

Br. J. Pharmacol.

Cannabis use amongst patients with inflammatory bowel disease

Eur. J. Gastroenterol. Hepatol.

Treatment of Crohn’s disease with cannabis: an observational study

Isr. Med. Assoc. J.

Impact of cannabis treatment on the quality of life, weight and clinical disease activity in inflammatory bowel disease patients: a pilot prospective study

Digestion

Marijuana use patterns among patients with inflammatory bowel disease

Inflamm. Bowel Dis.

Cannabis use provides symptom relief in patients with inflammatory bowel disease but is associated with worse disease prognosis in patients with Crohn’s disease

Inflamm. Bowel Dis.

Prevalence and patterns of marijuana use in young adults with inflammatory bowel disease

J. Pediatr. Gastroenterol. Nutr.

The effects of Delta-tetrahydrocannabinol and cannabidiol alone and in combination on damage, inflammation and in vitro motility disturbances in rat colitis

Br. J. Pharmacol.

Cannabidiol, a safe and non-psychotropic ingredient of the marijuana plant Cannabis sativa, is protective in a murine model of colitis

J. Mol. Med.

Topical and systemic cannabidiol improves trinitrobenzene sulfonic acid colitis in mice

Pharmacology

An orally active Cannabis extract with high content in Cannabidiol attenuates chemically-induced intestinal inflammation and hypermotility in the mouse

Front. Pharmacol.

The cannabinoid TRPA1 agonist cannabichromene inhibits nitric oxide production in macrophages and ameliorates murine colitis

Br. J. Pharmacol.

Effects of cannabinoids and cannabinoid-enriched Cannabis extracts on TRP channels and endocannabinoid metabolic enzymes

Br. J. Pharmacol.