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. Author manuscript; available in PMC: 2018 Nov 1.
Published in final edited form as: J Invest Dermatol. 2017 Jun 23;137(11):2445–2447. doi: 10.1016/j.jid.2017.05.035

Cannabinoid reduces inflammatory cytokines tumor necrosis factor alpha and type I interferons in dermatomyositis in vitro

Elizabeth S Robinson 1,2, Paul Alves 1,2, Muhammad M Bashir 1,2, Majid Zeidi 1,2, Rui Feng 3, Victoria P Werth 1,2
PMCID: PMC5651184  NIHMSID: NIHMS909075  PMID: 28652111

To the editor

Cannabis has been used in medicinal preparations for centuries to treat diseases such as chronic pain, neuropathy, and epilepsy. Recent research has identified anti-inflammatory properties in both endocannabinoids and phytocannabinoids. Our study investigated if the anti-inflammatory effects of a non-psychoactive cannabinoid called ajulemic acid could target the cytokines pathogenic in cutaneous dermatomyositis.

One obstacle to the use of cannabinoids as therapeutics is their psychoactive effect. It is well established that specific chemical modifications to tetrahydrocannabinol (THC) render it free of central nervous system activity. This strategy was employed to design ajulemic acid. Ajulemic acid has a 65-fold higher affinity for the cannabinoid 2 receptor compared to the cannabinoid 1 receptor (Tepper et al., 2014). The cannabinoid 2 receptor is found on immune cells and plays a natural role in resolving inflammation (Bouaboula et al., 1993, Munro et al., 1993). Ajulemic acid has a low affinity for the cannabinoid 1 receptor, which is found in the nervous system and mediates THC’s psychoactive effects. Preferential binding for the cannabinoid 2 receptor enables ajulemic acid to preserve THC’s anti-inflammatory properties without the psychological stimulatory effects, making it a potentially useful therapy for treating autoimmune diseases.

Prior studies of ajulemic acid in vitro with human blood and synovial cells demonstrated anti-inflammatory properties. Ajulemic acid reduced production of the inflammatory mediators interleukin-1β and interleukin-6, induced T cell apoptosis, and increased anti-inflammatory eicosanoids prostaglandin J2 and lipoxin A4 (Bidinger et al., 2003, Parker et al., 2008, Stebulis et al., 2008, Zurier et al., 2003, Zurier et al., 2009). Furthermore, multiple clinical trials of ajulemic acid administered orally found no serious adverse effects and no marijuana-like neurological effects (Corbus Pharmaceuticals’ clinical investigator brochure for JBT-101, version 1.11).

This study examined if the anti-inflammatory properties of ajulemic acid could target the pathogenic cytokines in cutaneous dermatomyositis, a disease with few effective and non-toxic therapies. The key inflammatory cytokines in dermatomyositis are type-I interferons (IFN) and tumor necrosis factor alpha (TNFα) (Kim et al., 2012, Nabatian et al., 2012, Pachman et al., 2000). We hypothesized that ajulemic acid could suppress production of these cytokines from immune cells of dermatomyositis patients in vitro.

After obtaining written, informed consent, peripheral blood mononuclear cells (PBMCs) were isolated from the blood of eighteen dermatomyositis patients. The cells were treated with 0, 3, 10 and 15 μM ajulemic acid (Corbus Pharmaceuticals Holdings, Norwood, MA, USA) for one hour and then cultured with lipopolysaccharide (LPS) for TNFα stimulation or CpG oligonucleotides (CpG) for IFN-α and IFN-β stimulation. After 18 hours, the supernatant was separated from the cells. TNFα, IFN-α and IFN-β production was quantified by enzyme-linked immunosorbent assay. Samples that did not produce baseline minimum cytokine levels without ajulemic acid were excluded from analysis. All expression values were log transformed before the analysis. The comparison between baseline and multi-dosage post-simulation gene expression was conducted though a linear mixed model, adjusted for within-subject correlations.

PMBC viability studies found that 98.2%, 97.4%, and 95.7% of cells were viable after incubation with 3, 10, and 15 μM ajulemic acid compared to 99.3% of cells incubated without ajulemic acid. Moderate and high ajulemic acid concentrations significantly suppressed TNFα secretion (Figure 1a), while all ajulemic acid doses significantly reduced IFN-α production (Figure 1b). Ajulemic acid suppressed TNFα secretion from LPS stimulated cells at concentrations of 10 μM (n=16) (p=0.0002) and 15 μM (n=14) (p<0.0001), but not at 3 μM (n=16) (p=0.4727) compared to cells not incubated with ajulemic acid (n=17). As opposed to untreated cells (n=8), ajulemic acid suppressed secretion of IFNα at all tested concentrations (3 μM n=7; 10 μM n=4; 15 μM n=3) (p≤0.0007). In the samples tested, all ajulemic acid doses decreased IFN-β production (Figure 1c).

Figure 1. Ajulemic acid suppressed TNFα, IFN-α, and IFN-β production from peripheral blood mononuclear cells of dermatomyositis patients.

Figure 1

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Ln(TNFα) (a), ln(IFN-α) (b) and ln(IFN-β) (c) secretion from peripheral blood mononuclear cells of dermatomyositis subjects treated with 0, 3, 10 and 15 μM ajulemic acid. Line represents the mean. Ajulemic acid (AJA); interferon (IFN); tumor necrosis factor alpha (TNFα).

During data analysis, we noted that some CpG stimulated PBMCs produced negligible levels of IFN-α even without ajulemic acid suppression. PBMCs from dermatomyositis patients on antimalarial therapies secreted significantly less IFN-α (n=5) compared to those not taking antimalarials (n=8) (p<0.0001), providing some insight into how antimalarials work in diseases like dermatomyositis (Figure 2). This result is consistent with prior findings that chloroquine and quinacrine suppress CpG-stimulated cytokine production (Kuznik et al., 2011).

Figure 2. IFN-α secretion was reduced in the peripheral blood mononuclear cells of subjects taking antimalarials.

Figure 2

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Ln(IFN-α) secretion from peripheral blood mononuclear cells of dermatomyositis subjects on antimalarials and dermatomyositis subjects not taking antimalarials. Line represents the mean. Interferon (IFN).

The role of cannabinoids in inflammation remains incompletely understood. Finding a safe and reliable way to benefit from the anti-inflammatory properties of cannabinoids may help many patients. Ajulemic acid, a non-psychoactive, orally administered cannabinoid, could offer such a treatment. The investigation presented here indicate that ajulemic acid, and theoretically other cannabinoids, limits production of the pathogenic cytokines in dermatomyositis. Phase II clinical trials are currently evaluating the effect of this innovative compound on disease morbidity and quality of life in dermatomyositis.

Footnotes

Ethical approval and consent to participate: This study was performed under approval from the University of Pennsylvania’s internal review board, the biohazard and safety subcommittee of the Philadelphia Veteran Affairs Hospital and the United States Drug Enforcement Administration.

Conflict of interest: Dr. Werth serves as a consultant to: Xoma, RPS, Janssen/Centocor, Biogen-Idec, Sanofi-Aventis, UVC, Novartis, Idera, Pfizer, Lupus Foundation of America, Medimmune, Rigel, Merck, Genentech, GSK Stiefel and Celgene. Dr. Werth holds stock ownership/options in UV Therapeutics, grants from the NIH, Amgen, Celgene, Rigel, Novartis, and the VA, and the copyright for the CDASI is owned by the University of Pennsylvania. This material is based upon work supported by the Department of Veterans Affairs (Veterans Health Administration, Office of Research and Development, Biomedical Laboratory Research and Development) and by the National Institutes of Health (NIH R21 AR066286) to VPW. Ajulemic acid was provided by Corbus Pharmaceuticals Holdings, Inc.

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