Synthesis and binding characteristics of tritiated TIPP[psi], a highly specific and stable delta opioid antagonist

The pseudopeptide H-Tyr-Tic psi [CH2-NH]Phe-Phe-OH (TIPP[psi]) is a delta opioid antagonist with high delta receptor affinity and unprecedented delta selectivity. TIPP[psi] was radiolabelled by catalytic tritiation of its precursor [Tyr(3',5'-I2)1]TIPP[psi]. The resulting radioligand, [3H]TIPP[psi], had a specific activity of 1.77 TBq/mmol (47.9 Ci/mmol) and showed high stability against enzymatic degradation. [3H]TIPP[psi] binding to rat brain membranes was saturable and Scatchard analysis indicated a single binding site with a Kd of 0.98 nM and a Bmax of 105.4 fmol/mg. A study of [3H]TIPP[psi] binding displacement by various receptor-selective opioids showed the expected rank order of potency (delta >> mu > kappa). [3H]TIPP[psi] represents an excellent new radioligand for delta receptor labelling studies in vitro and in vivo.