Differential antagonism of U69,593- and bremazocine-induced antinociception by (-)-UPHIT: Evidence of kappa opioid receptor multiplicity in mice

The effect of pretreatment with the kappa receptor nonequilibrium antagonist, (-)-UPHIT {1S,2S-trans-2-isothiocyanato-4,5-dichloro-N-methyl-N-[2-(1 -pyrrolidinyl)cyclohexyl]benzeneacetamide}, on U69,593 {(5α,7α,8β)-(-)-N-methyl-N-(7-(1-pyrrolidinyl) -1-oxaspiro(4,5)dec-8-yl)benzeneacetamide}- and bremazocine-induced antinociception was examined in mice. Both U69,593 and bremazocine produced antinociception in the warm water tail-flick test after i.c.v. administration. Pretreatment with the kappa antagonist, nor-binaltorphimine, at doses shown not to affect [D-Ala²,NMePhe⁴, Gly-ol]enkephalin-(mu-agonist) or [D-Pen², D-Pen⁵]enkephalin (delta-agonist)-induced antinociception, significanlly attenuated the effects of U69,593 and bremazocine, suggesting actions of these agonists at kappa receptors. Furthermore, β-funaltrxamine (mu antagonist) and ICI 174,864 {N,N,-diallyl-Tyr-(α-aminoisobutyric acid)₂-Phe-Leu-OH} (delta antagonist), had no effect on U69,593 or bremazocine in this test providing further evidence of kappa receptor-mediated activity. Pretreatment with (-)-UPHIT produced no effect alone and a long-lasting (up to 48 hr) antagonism of U69,593, but not bremazocine, antinociception. The antagonist actions of (-)-UPHIT did not alter the antinociceptive effects of [D-Ala²,NMePhe⁴, Glyol]enkephalin or [D-Pen², D-Pen⁵]enkephalin. These data suggest that (-)-UPHIT is a selective, long-lasting kappa antagonist which can differentially antagonize the antinocicieption produced by these two kappa agonists. These data provide evidence in vivo supportive of kappa receptor subtypes in the mouse, and suggest that (-)-UPHIT may be a useful probe for the exploration of kappa receptor heterogeneity.