Preclinical toxicity evaluation of JD5037, a peripherally restricted CB1 receptor inverse agonist, in rats and dogs for treatment of nonalcoholic steatohepatitis

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Highlights

  • Preclinical toxicology studies of JD5037, a peripheral CB1R inverse agonist, were conducted in rats and dogs.

  • JD5037 exhibited non-linear kinetics in both the species.

  • Fed state increased plasma exposure of JD5037 in dogs by 4.4–4.6-fold compared to fasting state.

  • NOAEL in rats was 150 mg/kg/day, while in male and female dogs it was 20 mg/kg/day and 75 mg/kg/day, respectively.

Abstract

JD5037 is a novel peripherally restricted CB1 receptor (CB1R) inverse agonist being developed for the treatment of visceral obesity and its metabolic complications, including nonalcoholic fatty liver disease and dyslipidemia. JD5037 was administered by oral gavage at 10, 40, and 150 mg/kg/day dose levels for up to 34 days to Sprague Dawley rats, and at 5, 20, and 75 mg/kg/day dose levels for 28 consecutive days to Beagle dogs. In rats, higher incidences of stereotypic behaviors were observed in 10 mg/kg females and 40 mg/kg males, and slower responses for reflex and sensory tests were observed only in males at 10 and 40 mg/kg during neurobehavioral testing. Sporadic minimal incidences of decreased activity (males) and seizures (both sexes) were observed in rats during daily clinical observations, without any clear dose-relationship. Male dogs at 75 mg/kg during treatment period, but not recovery period, had an increased incidence of gut associated lymphoid tissue hyperplasia and inflammation in the intestine. In both species, highest dose resulted in lower AUCs indicative of non-linear kinetics. Free access to food increased the plasma AUC by ~4.5-fold at 20 mg/kg in dogs, suggesting presence of food may help in systemic absorption of JD5037 in dogs. Based on the study results, 150 mg/kg/day in rats, and 20 and 75 mg/kg/day doses in male and female dogs, respectively, were determined to be the no-observed-adverse-effect-levels (NOAELs).

Introduction

Nonalcoholic fatty liver disease (NAFLD) is an umbrella term for a variety of ailments associated with fat accumulation in the liver of people who drink little or no alcohol. NAFLD is one of the most common liver diseases occurring in people in their 40s–50s, mainly in the Western countries. About 20–30% of the adult population and 10% of children in Western countries are estimated to be suffering from NAFLD (Sanyal et al., 2015). Nonalcoholic steatohepatitis (NASH) is the most severe form of NAFLD, characterized by fat accumulation and inflammation in the liver which subsequently may result in liver cirrhosis. Currently there are no effective drugs available for treatment of NASH. Several drugs targeting various signaling pathways, including lipid metabolism, glucose metabolism, inflammation, fibrosis and apoptosis are under investigation for NASH therapy (Wattacheril et al., 2018). The endocannabinoid system plays a central role in regulation of food intake, lipid metabolism and energy expenditure in the body, mediated by G-protein coupled CB1 receptors (CB1R) (Kunz et al., 2008). CB1R are expressed in the brain as well as in the peripheral organs such as the liver, muscles, kidney, adipose tissues, gastrointestinal tract, thyroid, pancreas, and adrenals (Kirilly et al., 2012). Induction of CB1R has been associated with increased food consumption leading to obesity. Hence, research attention was focused on developing CB1R antagonists for the treatment of lipid disorders such as NASH, obesity, and metabolic syndrome.

Rimonabant, the first approved inverse agonist of CB1R showed reduction in food consumption, improved lipid profile, and reduced visceral and hepatic fat (Carai et al., 2006). However, due to its brain penetrant ability, neurologic and psychiatric effects like dizziness, seizures, depression, suicidal tendency, insomnia, and headache were observed (Sam et al., 2011). These observations resulted in withdrawal of rimonabant from the European and Brazilian markets in 2008. However, recent studies revealed that desired pharmacological effects on lipid control without having psychiatric and neurological side effects can be achieved by selectively blocking peripheral CB1R (Chen et al., 2017; Chorvat, 2013; Hsiao et al., 2015; Shrestha et al., 2018; Tam et al., 2012). Hence, an inverse agonist of selective peripheral CB1R, JD5037, was developed jointly by United States National Institute of Health and Jenrin Discovery (Tam et al. 2012, 2017). JD5037 exhibited minimal to no (below level of detection) brain penetration in chow-fed mice (Chorvat et al., 2012; Tam et al., 2012). JD5037 showed zero percent occupancy at CB1R in mouse brain, while rimonabant (a brain penetrant CB1R inverse agonist) showed 87% occupancy at 30 mg/kg dose (Chorvat et al., 2012). JD5037 has been found effective in reducing appetite, body weight, hepatic steatosis and improving insulin resistance in diet induced obese mice (Chorvat et al., 2012; Cinar et al., 2014; Tam et al. 2012, 2017). JD5037 has been now permitted by the U.S.-FDA for Phase-1 clinical trials for NASH indication in the United States, for which JD5037 has been licensed by Corbus Pharmaceuticals as CRB-4001. Here, we present the results of preclinical toxicology studies of JD5037 in Sprague Dawley rats and Beagle dogs that were conducted to support an Investigational New Drug (IND) application submission to U.S.-FDA. The purpose of these studies was to evaluate toxicity resulting from repeat dose administration of JD5037 and possible reversal of toxicity findings after the recovery period. All the studies reported here were conducted in accordance with the principles of U.S.-FDA Good Laboratory Practice guideline (21 CFR, Part 58).

Section snippets

Experimental design

Sprague Dawley rats were obtained from Charles River Laboratories, Raleigh, NC (Charles River Nomenclature: CD® rats). The rats were approximately 8–9 weeks of age. The males weighed 286.1–344 g and the females weighed 192.6–236.5 g at the initiation of dose administration.

Beagle dogs were obtained from Covance, Cumberland, VA. The dogs were approximately 6 months of age at the initiation of dose administration. The males weighed 8.21–9.92 kg and the females weighed 6.57–9.86 kg at the

Dose formulation analysis

All the formulations (1, 4, and 15 mg/mL concentrations) were homogeneous. The relative standard deviation (RSD), which indicates homogeneity of the formulations, was within 0.2–4.1% and 0.4–0.8% in the rat and dog study, respectively. The formulation concentrations were also within acceptable limit. Relative error for all formulations was within 0 to −5.9% (rat) and −1.8 to −6.2% (dog) of the target concentration (RE, relative error) at all the concentration levels. The formulations were

Discussion

The preclinical toxicology studies of JD5037 were conducted in rats and dogs. In dogs, the observed increase in the toxicokinetic parameters such as clearance and volume of distribution as the dose increased indicates non-linear kinetics of JD5037. The dose-normalized Cmax (Cmax/dose) and AUClast (AUClast/dose) values were decreased with dose increment in both sexes of both the species. The absolute Cmax or AUClast values did not increase dose proportionately. Additionally, the elimination

Conclusion

While gastrointestinal effects were observed in male dogs in this study, these appear to be either completely or partially reversible with discontinuation of treatment. Since these effects were not observed in female dogs and none of the rats, the possibility of translatability of this nonclinical finding to humans seems feeble. It will be worthwhile to consider non-linear kinetics of JD5037 observed in nonclinical studies during a Phase 1 clinical trial preparation. A special consideration

Disclosure

The abstract and poster of this research work have been presented at Society of Toxicology (2018) meeting.

Funding body information

This research project has been funded in whole with Federal funds from the National Center for Advancing Translational Sciences, National Institutes of Health, USA (Contract No. HHSN261200800001E), under BrIDGs/NCATS program. Manuscript preparation and publishing cost was funded by Battelle Memorial Institute.

Declaration of competing interest

The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: R. Chorvat is an employee of Jenrin Discovery. JD5037 is being developed by Corbus Pharmaceuticals that licensed the rights from Jenrin Discovery.

Acknowledgements

This research project has been funded in whole with Federal funds from the National Center for Advancing Translational Sciences, National Institutes of Health, USA (Contract No. HHSN261200800001E) under BrIDGs/NCATS program. The content of this publication does not necessarily reflect the views of policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government.

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