Therapeutic Pipeline

The mouse model of inflammatory bowel disease (IBD) induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS), which mimics Crohn’s disease and colitis, was used to evaluate the therapeutic efficacy of candidate compounds compared to CBG (labeled DM100) and a positive control Clyclosporin A. Based on the total colonic damage score (Figure 1), disease activity index, colon weight/length, and histopathology (not shown), we selected DM350-P, and DM300 for further testing.

Figure 1: Total colonic macroscopic lesion score in the TNBS model: a composite of adhesions, strictures, ulcers/inflammation, and wall thickness. The positive control, DM350-P and DM300 groups all showed significant improvements over vehicle; while CBG had no effect. The positive control Cyclosporin A is not commonly used for IBD, an immunosuppressant with known side effects. †p< 0.05, vs sham control; *p< 0.05, vs vehicle control; two-way ANOVA followed by Bonferroni test

The NSAID (Non-steroidal anti-inflammatory drugs) indomethacin, induces lesions in the small bowel resembling Crohn’s disease. Both DM300 and DM350-P demonstrated significant improvement in all disease parameters, including small intestine weight (Figure 2), gross score, and histopathology. Unfortunately, rats treated with CBG and DM200 experienced severe toxicity. Subsequent analysis confirmed Cyclooxygenase (COX) inhibition activity of these molecules, which may have exacerbated the NSAID-induced injury. No COX inhibition was observed in assays with DM300 or DM350-P (data not shown).

Figure 2: Small intestine weight is a simple and reliable measure of anti-inflammatory activity in the rat indomethacin-induced Crohn’s model. Both DM300 and DM350-P treatment result in significant improvement over vehicle. Notably at as low as 10 mg/kg DM350-P is close to as potent as Dexamethasone, a steroid with known side-effects. †p ≤ 0.05 ANOVA (Dunnett’s post-hoc) vs. Vehicle

Given CBD’s robust efficacy in rodent epilepsy models and human clinical trials, we screened DM300 and DM350-P for anti-convulsant activity in the rat PTZ seizure model. DM350-P showed lower brain exposure, resulting in significant but modest improvements (data not shown). At the lowest doses, DM300 completely eliminated severe tonic-clonic seizures in a rat model of PTZ-induced seizures. Comparable results were observed in CBD- treated rats. (Figure 3). Valproate, a popular anti-convulsive with known side effects was also tested for comparison.

DM300 demonstrates improved brain exposure compared to CBD and does not lead to any markers of liver injury. In contrast, at similar exposure levels, CBD was found to increase total bilirubin in rodents. The inhibition of the bile salt export pump (BSEP) has emerged as a significant mechanism that may contribute to increased bilirubin and the initiation of human drug-induced liver injury. In vitro studies suggested CBD inhibits BSEP while DM300 has no effect (not shown). The efficacy and lack of toxicity suggest that DM300 could be an adjuvant treatment in combination with CBD or Valproate to enhance efficacy at lower doses.

Figure 3: Effect of DM300, CBD and Valproate on PTZ (75 mg/kg s.c.) induced seizures in Sprague Dawley rats. Latency to severe tonic-clonic seizure with loss of righting reflex (stage 5), right. Data was analyzed by Kruskal Wallis test followed by Dunn’s post-hoc test *p<0.05.

Utilizing advanced formulation techniques, Demeetra has developed a method to produce solid tablet prototypes for cannabinoids. These solid formulations demonstrate increased drug load and bioavailability compared to a sesame seed oil formulation similar to Epidiolex. Additionally, solid cannabinoid tablets bolster our IP portfolio, are expected to reduce manufacturing costs, and enable more advanced delivery options, such as delayed release.

Figures 4 + 5: Representative images of two different solid formulations in prototype tablet form. Solid formulations can reach 50% drug load.