title: NCS-MPP (4-(2'-methoxy-phenyl)-1-[2'-(N-2"-pyridyl)-p-isothiocyanobenz amido]-ethyl-piperazine): a high affinity and irreversible 5-HT1A receptor ligand.
abstract: A novel irreversible 5-HT1A receptor binding ligand, NCS-MPP (4-(2'- methoxy-phenyl)-1-[2'-(N-2"-pyridyl)-p-isothiocyanobenzamido]- ethyl-piperazine), based on the new 5-HT1A receptor antagonist p-MPPI (4-(2'-methoxy-phenyl)-1-[2'-(N-2"-pyridyl)-p-iodobenzamido]-ethyl -piperazine ), was synthesized, and its binding characteristics were evaluated using in vitro homogenate binding with rat hippocampal membranes. The Ki value of NCS-MPP was estimated to be 1.8 +_ 0.2 nM using analysis of concentration-dependent inhibition for the binding of [125I]p-MPPI to 5-HT1A receptors. NovaScreen of NCS-MPP showed low to moderate binding affinities to alpha-1, alpha-2-adrenergic and 5-HT2 receptors, with Ki values of 350, 420, and 103 nM, respectively. These data strongly suggest that the ligand bound to 5-HT1A receptors with high affinity and high selectivity. Irreversible inhibition of [125I]p-MPPI binding by NCS-MPP following a 5 min incubation at room temperature was concentration dependent; the inhibition increased to 50% at a concentration less than 10 nM, and became more pronounced (90%) at 400 nM. Under similar assay conditions, NCS-MPP was significantly less efficient in irreversibly inhibiting agonist ligand [125I]8-OH-PIPAT binding to 5-HT1A receptors at lower concentrations (<10nM). After pretreatment of membranes with a low concentration of NCS-MPP (2nM), there was an apparent loss of [125I]p-MPPI binding sites, as expected, but no change in the binding affinity (Kd) was observed. However, the significant increase in Kd at a higher concentration of NCS-MPP (50 nM) indicated that there may be a secondary alkylation site, which may not be directly involved in p-MPPI binding to receptors; nevertheless, it would lead to an increased Kd value. The availability of an irreversible ligand, NCS-MPP, may provide a useful tool for studies of 5-HT1A receptors in the central nervous system.
authors.affiliation: Department of Radiology, University of Pennsylvania, Philadelphia 19104, USA.
authors.forename: M P
authors.initials: MP
authors.lastname: Kung
authors.forename: M
authors.initials: M
authors.lastname: Mu
authors.forename: Z P
authors.initials: ZP
authors.lastname: Zhuang
authors.forename: H F
authors.initials: HF
authors.lastname: Kung
mesh_terms: D000818:Animals; D001549:Benzamides / Q000378:metabolism* / Q000494:pharmacology*; D006624:Hippocampus / Q000187:drug effects; D017879:Isothiocyanates / Q000378:metabolism* / Q000494:pharmacology*; D007700:Kinetics; D015394:Molecular Structure; D011869:Radioligand Assay; D051381:Rats; D011985:Receptors, Serotonin / Q000187:drug effects*; D044263:Receptors, Serotonin, 5-HT1
chemical_list: C098477:4-(2'-methoxyphenyl)-1-((2'-(N-2''-pyridyl)-4-isothiocyanobenzamido)ethyl)piperazine; D001549:Benzamides; D017879:Isothiocyanates; D011985:Receptors, Serotonin; D044263:Receptors, Serotonin, 5-HT1