BACKGROUND AND OBJECTIVE

Renzapride (ATL-1251), a novel benzamide, is currently under clinical development for the treatment of irritable bowel syndrome (IBS). Previous in vitro and in vivo experimental studies have characterized renzapride as a full serotonin 5-HT(4) receptor agonist on the gut and a 5-HT(3) receptor antagonist. Clinical studies have confirmed the therapeutic efficacy, tolerability and safety of renzapride in patients with constipation-predominant IBS. This study set out to characterize the pharmacological profile of renzapride and its potential metabolic products at both 5-HT and other monoamine receptors in the gut.

METHODS

The affinity of renzapride, its (+) and (-) enantiomers, and its primary metabolite, renzapride N-oxide and its enantiomers, for serotonin receptors was assessed by means of in vitro radioligand binding inhibition studies. After membranes prepared from animal tissue or membranes of cell lines transfected with cloned human receptors had been incubated with radiolabelled ligand with high affinity for a specific receptor, renzapride was added to competitively inhibit this binding. Levels of bound radioligand were measured by filtration and counting of the bound radioactivity. In instances where >50% inhibition of radioligand binding had occurred, the inhibition constant (K(i)) was calculated. Metabolism of renzapride by liver microsomes was assessed by incubating 10 micromol/L renzapride with human liver microsome samples for 60 minutes at 37 degrees C. After the reaction was stopped, the samples were centrifuged and the supernatant analysed for metabolites by high-pressure liquid chromatography (HPLC). The potential inhibitory effects of renzapride on cytochrome P450 (CYP) enzymes were assessed by incubating renzapride at various concentrations over a 1-500 micromol/L concentration range with microsomes genetically engineered to express a single CYP.

RESULTS

Renzapride was selective for serotonergic receptors and, in particular, had high affinity for human 5-HT(3) and guinea-pig 5-HT(4) receptors (K(i) 17 and 477 nm, respectively). Inhibitory properties at 5-HT(2B) receptors were also identified for renzapride, as well as some affinity for 5-HT(2A) and 5-HT(2C) receptors. Renzapride N-oxide and its enantiomers demonstrated much lower affinity for all 5-HT receptors compared with renzapride. Renzapride was metabolized by liver microsomes to a limited extent and there was no significant non-microsomal metabolism of renzapride. Renzapride did not inhibit the major CYP drug-metabolizing enzymes CYP2C9, CYP2D6, CYP1A2, CYP2A6, CYP2C19, CYP2E1 or CYP3A4 at concentrations consistent with use in a clinical setting.

CONCLUSIONS

These results confirm and extend earlier studies in animal and human receptors that show renzapride is a potent and generally full 5-HT(4) receptor agonist and 5-HT(3) receptor antagonist. The results reported in the present study indicate that the metabolites of renzapride are minor and are unlikely to contribute to its therapeutic profile or lead to interaction of renzapride with other drugs that inhibit the major drug-metabolizing enzymes in the liver at therapeutic doses. These data contribute to the understanding of the pharmacological actions and metabolic fate of renzapride in vivo.