Oral repeated-dose toxicity studies of BIA 10-2474 in CD-1 mice

Abstract

We independently and retrospectively reviewed three studies that evaluated the toxicity of BIA 10-2474 (3-(1-(cyclohexyl(methyl)carbamoyl)-lH-imidazol-4-yl)pyridine 1-oxide), a novel fatty acid amide hydrolase (FAAH) inhibitor in male and female CD-1 mice based upon raw data obtained from Bial Portela & Companhia S.A. (São Mamede do Coronado, Portugal). These studies were carried out prior to the clinical trial with BIA 10-2474 and formed part of the regulatory submission. An initial oral dose range-finding study with BIA 10-2474 showed that doses from 600 mg/kg/day were poorly tolerated with a high mortality rate and signs of weakness, prostration, labored breathing, clear lacrimation, tachypnea/bradypnea and decreased activity. At lower doses (100 and 300 mg/kg/day) there were few signs but post-mortem analysis showed increased liver weight. In a 28-day study a third of the animals receiving 500 mg/kg/day died or required euthanasia, with similar signs to those seen in the dose-range finding study. At lower doses (i.e. 100 and 300 mg/kg/day) there were few clinical signs although there were dose-related decreases in erythrocyte count and hemoglobin. Histopathology was seen in the 300 and 500 mg/kg/day groups and included hepatocellular hypertrophy (with increased liver weight), nephropathy and enterocyte vacuolation. Finally, in the 13-week oral gavage study, BIA 10-2474 was administered to CD-1 mice of both sexes at dose levels of 25, 75 and 150 mg/kg/day. Under these conditions, there were almost no clinical signs apart from a tendency to increase body-weight. Cholesterol was increased at 75 and 150 mg/kg and remained high after recovery. Liver and spleen weights increased at 75 and 150 mg/kg/day. Histopathologically, there was a dose-dependent increase in sciatic nerve and myofiber degeneration, hepatocellular hypertrophy, nephropathy and inflammatory loci in the bladder. The nerve damage and nephropathy seen at 150 mg/kg/day persisted after a 4-week recovery period. Toxicokinetic analysis in the 4- and 13-week studies showed that exposure was broadly dose-proportional with no evidence of accumulation. On the basis of the changes seen during the 13-week study, the NOAEL was established at 75 mg/kg/day.

Keywords: BIA 10–2474; CD-1 mice; Endocannabinoid system; Endogenous anandamide; Toxicology.