title: Pharmacological analysis of the novel, rapid, and potent inactivation of the human 5-Hydroxytryptamine7 receptor by risperidone, 9-OH-Risperidone, and other inactivating antagonists.
abstract: In a previous publication, using human 5-hydroxytryptamine(7) (h5-HT(7)) receptor-expressing human embryonic kidney (HEK) 293 cells, we reported the rapid, potent inactivation of the h5-HT(7) receptor stimulation of cAMP production by three antagonists: risperidone, 9-OH-risperidone, and methiothepin (Smith et al., 2006). To better understand the drug-receptor interaction producing the inactivation, we 1) expanded the list of inactivating drugs, 2) determined the inactivating potencies and efficacies by performing concentration-response experiments, and 3) determined the potencies and efficacies of the inactivators as irreversible binding site inhibitors. Three new drugs were found to fully inactivate the h5-HT(7) receptor: lisuride, bromocryptine, and metergoline. As inactivators, these drugs displayed potencies of 1, 80, and 321 nM, respectively. Pretreatment of 5-HT(7)-expressing HEK cells with increasing concentrations of the inactivating drugs risperidone, 9-OH-risperidone, methiothepin, lisuride, bromocriptine, and metergoline potently inhibited radiolabeling of the h5-HT(7) receptor, with IC(50) values of 9, 5.5, 152, 3, 73, and 10 nM, respectively. We were surprised to find that maximal concentrations of risperidone and 9-OH-risperidone inhibited only 50% of the radiolabeling of h5-HT(7) receptors. These results indicate that risperidone and 9-OH risperidone may be producing 5-HT(7) receptor inactivation by different mechanisms than lisuride, bromocryptine, metergoline, and methiothepin. These results are not interpretable using the conventional model of G-protein-coupled receptor function. The complex seems capable of assuming a stable inactive conformation as a result of the interaction of certain antagonists. The rapid, potent inactivation of the receptor-G-protein complex by antagonists implies a constitutive, pre-existing complex between the h5-HT(7) receptor and a G-protein.
authors.affiliation: Center for Neuropharmacology and Neuroscience, Albany Medical College, 47 New Scotland Avenue, Albany, NY 12208. teitlem@mail.amc.edu.
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Knight
authors.forename: Carol
authors.initials: C
authors.lastname: Smith
authors.forename: Nicole
authors.initials: N
authors.lastname: Toohey
authors.forename: Michael T
authors.initials: MT
authors.lastname: Klein
authors.forename: Milt
authors.initials: M
authors.lastname: Teitler
mesh_terms: D017371:8-Hydroxy-2-(di-n-propylamino)tetralin / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D006801:Humans; D007555:Isoxazoles / Q000494:pharmacology*; D000068882:Paliperidone Palmitate; D011743:Pyrimidines / Q000494:pharmacology*; D011985:Receptors, Serotonin / Q000187:drug effects*; D018967:Risperidone / Q000494:pharmacology*; D012702:Serotonin Antagonists / Q000494:pharmacology*
chemical_list: D007555:Isoxazoles; D011743:Pyrimidines; D011985:Receptors, Serotonin; D012702:Serotonin Antagonists; C104425:serotonin 7 receptor; D017371:8-Hydroxy-2-(di-n-propylamino)tetralin; D018967:Risperidone; D000068882:Paliperidone Palmitate