LNP 906, the first high-affinity photoaffinity ligand selective for I1 imidazoline receptors

1 The hypotensive effect of imidazoline-like drugs, such as clonidine, was attributed both to alpha(2)-adrenergic receptors and nonadrenergic imidazoline receptors, which are divided into I-1, I-2 and I-3 subtypes. 2 We have recently synthesized a derivative of (2-(2-chloro-4-iodo-phenylamino)-5-methyl-pyrroline (LNP 911), the first high-affinity and selective ligand for I-1 receptors (I1R), with a photoactivable function (LNP 906). 3 This work aims to test whether this derivative retained the binding properties of LNP 911 and bound irreversibly to I1R. 4 Binding studies showed that LNP 906 exhibited nanomolar affinity for I1R and was selective for I1R over I-2 receptors and alpha(2)-adrenergic receptors (alpha(2)Ars). 5 Upon exposure to u.v. light, LNP 906 irreversibly blocked the binding of [I-125]-paraiodoclonidine (PIC) to I1R, time- and dose-dependently, on PC12 cell membranes and interacted with I1R in a reversible and competitive manner in the absence of light. Pharmacological studies showed that this blockade was prevented by the concomitant presence of rilmenidine (a well-known I-1 agonist), but not by rauwolscine (an alpha(2) antagonist). 6 Finally, LNP 906 clearly antagonized the decrease in forskolin-stimulated cAMP level induced by rilmenidine, but not by melatonin. 7 These results indicate that LNP 906 is the first high-affinity and selective photoaffinity ligand for I1R and that it behaves as an I1R antagonist.