3-Hydroxyanthranilic acid accumulation following administration of the 3-hydroxyanthranilic acid 3,4-dioxygenase inhibitor NCR-631

Abstract

In the kynurenine pathway of tryptophan metabolism, 3-hydroxyanthranilic acid is the substrate for formation of the excitotoxin quinolinic acid by 3-hydroxyanthranilic acid 3,4-dioxygenase. This study was designed to characterize the effects on 3-hydroxyanthranilic acid after treatment with the 3-hydroxyanthranilic acid 3,4-dioxygenase inhibitor 4,6-di-bromo-3-hydroxyanthranilic acid (NCR-631) in Sprague–Dawley rats. The blood plasma and brain concentrations of 3-hydroxyanthranilic acid were found to increase rapidly in a dose-dependent manner after gavage administration of NCR-631. However, the effect was relatively transient, with a decline in 3-hydroxyanthranilic acid levels already at 1h after NCR-631 treatment. Similar increases in plasma levels of 3-hydroxyanthranilic acid were observed following either gavage or parenteral (i.v. or s.c.) administration of NCR-631 (25 mg/kg). Only a minor enhancement of the NCR-631-induced increase in plasma 3-hydroxyanthranilic acid levels was found after sub-chronic treatment (25 mg/kg by gavage; 7 days, b.i.d.), suggesting a low propensity for altered 3-hydroxyanthranilic acid 3,4-dioxygenase activity following repeated inhibition. Administration of [ 14C]NCR-631 suggested 20 min initial plasma half life and an oral absorption around 50%. A dose of 250 mg/kg [ 14C]NCR-631 given by gavage provided plasma levels of almost 2 μmol/ml and a brain concentration of approximately 16 nmol/g, when analyzed 15 min after administration. Neither acute nor sub-chronic administration of NCR-631 caused any substantial effects on quinolinic acid levels in plasma or brain. Also, the plasma levels of kynurenic acid, another neuroactive kynurenine pathway metabolite, were unaffected by acute NCR-631 treatment. Moreover, the brain levels of the major cerebral tryptophan metabolites 5-hydroxytryptamine and 5-hydroxyindoleacetic acid remained unchanged following administration of NCR-631. Although reversible inhibition of 3-hydroxyanthranilic acid 3,4-dioxygenase with NCR-631 in normal rats is insufficient to cause substantial changes in the levels of quinolinic acid or other important tryptophan metabolites, it causes a major accumulation of the substrate 3-hydroxyanthranilic acid.