title: Synthesis and characterization of an affinity label for brain receptors to psychotomimetic benzomorphans: differentiation of sigma-type and phencyclidine receptors.
abstract: Brain sigma-type receptors and phencyclidine receptors are thought to mediate the psychotomimetic effects of benzomorphans and phencyclidine in humans. Recently, we reported the characterization of a selective sigma receptor ligand, 1,3-di-o-tolyl-guanidine (DTG), that shows negligible crossreactivity with phencyclidine receptors. Here we describe the synthesis and characterization of an isothiocyanate derivative of DTG, di-o-tolyl-guanidine-isothiocyanate (DIGIT). Guinea pig brain membranes treated with nanomolar doses of DIGIT followed by extensive washing exhibit a dose dependent reduction of [3H]DTG and (+)[3H]3-(3-hydroxyphenyl)-N-(1-propyl)piperidine [+)[3H]3-PPP) binding to sigma receptors. Binding of radiolabelled ligands for phencyclidine, mu-opioid, benzodiazepine and dopamine-D2 receptors is not affected by membrane treatment with DIGIT, indicating specificity of the affinity label for sigma-type receptors. Treatment of DIGIT-derivatized membranes with 2 M NaCl does not result in recovery of sigma binding activity, suggesting that DIGIT's interaction with sigma receptors is not of an ionic nature. Equilibrium saturation binding experiments show that the inhibition of [3H]DTG binding to sigma receptors by DIGIT pretreatment of membranes is attributable to an irreversible reduction in the affinity (increase in Kd) of sigma receptors for DTG. The finding that sigma receptors are irreversibly modified by DIGIT whereas phencyclidine receptors are not affected suggests that sigma receptors are physically separate from phencyclidine receptors. The availability of a selective affinity label for the sigma receptor should facilitate the purification of the receptors and the characterization of sigma-type pharmacological effects in vivo and in vitro.
authors.affiliation: Vollum Institute for Advanced Biomedical Research, Oregon Health Sciences University, Portland 97201.
authors.forename: J T
authors.initials: JT
authors.lastname: Adams
authors.forename: P M
authors.initials: PM
authors.lastname: Teal
authors.forename: M S
authors.initials: MS
authors.lastname: Sonders
authors.forename: B
authors.initials: B
authors.lastname: Tester
authors.forename: J S
authors.initials: JS
authors.lastname: Esherick
authors.forename: M W
authors.initials: MW
authors.lastname: Scherz
authors.forename: J F
authors.initials: JF
authors.lastname: Keana
authors.forename: E
authors.initials: E
authors.lastname: Weber
mesh_terms: D000345:Affinity Labels / Q000138:chemical synthesis* / Q000494:pharmacology; D000818:Animals; D001575:Benzomorphans / Q000378:metabolism*; D001667:Binding, Competitive; D001921:Brain / Q000378:metabolism*; D006168:Guinea Pigs; D006213:Hallucinogens / Q000378:metabolism*; D066298:In Vitro Techniques; D008297:Male; D009019:Morphinans / Q000378:metabolism*; D017981:Receptors, Neurotransmitter / Q000378:metabolism*; D011957:Receptors, Opioid / Q000378:metabolism*; D017478:Receptors, Phencyclidine; D017480:Receptors, sigma
chemical_list: D000345:Affinity Labels; D001575:Benzomorphans; D006213:Hallucinogens; D009019:Morphinans; D017981:Receptors, Neurotransmitter; D011957:Receptors, Opioid; D017478:Receptors, Phencyclidine; D017480:Receptors, sigma