Correlation between binding characteristics and functional antagonism in human glioma cells by tachykinin NK1 receptor antagonists

Binding characteristics and functional antagonism exerted by two structurally related tachykinin NK1 receptor antagonists, MEN 11467 ((1R,2S)-2N[1(H)indol-3-yl-carbonyl]-1-N-{Nalpha(p-tolylacetyl+ ++)-Nalpha(methyl)-D-3-(2-naphthyl)alanyl}diaminocyclohexane) and FK888 (N2-[(4R)-4-hydroxy-1-(1-methyl-1H-indol-3-yl)carbonyl-L-prolyl]-N-methy l-N-phenylmethyl-L-3-(2-naphthyl)alaninamide), were investigated in the human astrocytoma cell line U373 MG. In radioligand binding studies, conducted with [3H]substance P and intact cells at 37 degrees C, MEN 11467 bound to tachykinin NK1 receptors in an irreversible manner with a Ki value of 1.2+/-0.5 nM while FK888 bound in competitive manner with a Ki value of 4.6+/-2.2 nM. Receptor blockade by both antagonists resulted in a powerful and complete inhibition of functional responses induced by substance P, such as accumulation of the second messenger inositol monophosphate or interleukin-6 release. However, MEN 11467 showed a greater potency for blocking functional responses than FK888 despite their similar affinity for human tachykinin NK1 receptors. Moreover, MEN 11467 was more potent in inhibiting late rather than early phases of substance P-induced inositol monophosphate accumulation and its antagonism was enhanced by drug preincubation and barely affected by removal of unbound drug from the external medium, suggesting that MEN 11467 bound in a tight manner to the receptor. Such behaviour was not observed with the competitive and rapidly reversible antagonist FK888. These data indicate that the small differences in the chemical structure of MEN 11467 and FK888 determine the different binding characteristics at the tachykinin NK1 receptor and which are responsible for the greater potency of MEN 11467 for blocking functional responses. The Ki value obtained in binding studies may be inadequate to reveal the real potency of tachykinin NK1 receptor antagonists.