Characterization of ibodutant at NK2 receptor in human colon

We have characterized the pharmacological profile of the nonpeptide tachykinin NK2 receptor antagonist ibodutant (MEN15596) through radioligand binding and contractility assays in the human colon smooth muscle. The antagonist affinity of ibodutant was evaluated through concentration-dependent inhibition curves at the [I-125]NKA specific binding by using membranes prepared from human colon smooth muscle. In this assay the affinity of ibodutant (pK(i) 9.9) was compared to that of other two selective NK2 receptor antagonists, nepadutant (pK(i) 8.4) and saredutant (pK(i) 9.2). The antagonist potency of ibodutant was evaluated towards the [beta Ala(8)]NKA(4-10)-mediated contractions of human colon smooth muscle strips. In this assay ibodutant (3, 10, 30 and 100 nM) induced a concentration-dependent rightward shift of the [beta Ala(8)]NKA(4-10) concentration-response curves without depressing the maximal contractile effect. The analysis of the curves yielded a Schild-plot linear regression with a slope not different from unity (1.02), thus indicating a surmountable antagonist behavior. The calculated apparent antagonist potency as pK(B) value was 9.1. No sex related differences were observed in NK2 receptor pharmacology for [beta Ala(8)]NKA(4-10) or ibodutant in colonic strips obtained from male or female patients. Reversibility experiments of tachykinin NK2 receptor blockade indicated that the inhibition of the agonist-induced contractions in preparations pre-exposed to ibodutant, and afterwards subjected to repeated washing cycles remained almost constant showing no sign of recovery during the 3 h observation period. Overall, the present study indicates ibodutant as a potent tachykinin NK2 receptor antagonist in the human colon tissue, also endowed with a persistent duration of action. (C) 2013 Elsevier B.V. All rights reserved.