title: Heterogeneity of binding sites for ICI 198,615 in human lung parenchyma.
abstract: We have identified and characterized two different subclasses of binding site for the novel peptido-leukotriene (LT) antagonist, [3H]ICI 198,615, in membranes from human lung parenchyma using a receptor-ligand assay. This novel compound is representative of a new class of LT receptor antagonists and it has been demonstrated to be several orders of magnitude more potent and selective than most other LT antagonists described to date. The binding of [3H]ICI 198,615 is rapid, specific and saturable. Equilibrium was reached within 5-10 min. Non linear fitting of dissociation time courses has revealed the presence of two different components (K(off)1 = 8.3 +/- 6.8 x 10(-4) sec-1 and K(off)2 = 0.79 +/- 1.66 x 10(-3) sec-1) of the kinetic curves, suggesting heterogeneity of the binding sites. Computer analysis of equilibrium binding data obtained at 25 degrees results in a model with two classes of binding sites, a high affinity-low capacity class with Kd1 = 0.024 +/- 0.014 nM and Bmax1 = 0.015 +/- 0.004 pmol/mg protein and a low affinity-high capacity class with Kd2 = 6326 +/- 3859 nM and Bmax2 = 473 +/- 383 pmol/mg protein. In competition studies, LTD4 was also found to interact with two classes of binding site (Kd1 = 0.016 +/- 0.008 nM and Kd2 = 15195 +/- 8965 nM). On the contrary, LTE4 and LTC4 were found to interact with a homogeneous class of sites only with Kd = 7466 +/- 4629 nM and Kd = 428 +/- 73 nM, respectively. Furthermore, we have evaluated the effect of a number of LT antagonists on the binding of [3H]ICI 198,615. Ro 24-5913 (Kd = 3.0 +/- 2.1 nM), FPL55712 (Kd = 4945 +/- 2868 nM), LY171883 (Kd = 19628 +/- 12365 nM), SKF 104353 (Kd = 74.2 +/- 46 nM) and its enantiomer SKF 104373 (Kd = 13627 +/- 6813 nM) were found to interact with a single class of binding sites. The present studies indicate a heterogeneity of binding sites for ICI 198,615 in membranes from human lung parenchyma and that ICI 198,615 is a very potent and selective antagonist of LTD4 receptors in this tissue.
authors.affiliation: Laboratory of Molecular Pharmacology, University of Milan, Italy.
authors.forename: G E
authors.initials: GE
authors.lastname: Rovati
authors.forename: S
authors.initials: S
authors.lastname: Giovanazzi
authors.forename: M
authors.initials: M
authors.lastname: Mezzetti
authors.forename: S
authors.initials: S
authors.lastname: Nicosia
mesh_terms: D001249:Asthma / Q000188:drug therapy / Q000378:metabolism; D001665:Binding Sites; D002867:Chromones / Q000494:pharmacology; D003998:Dicarboxylic Acids / Q000494:pharmacology; D006801:Humans; D007191:Indazoles / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000494:pharmacology; D007700:Kinetics; D008168:Lung / Q000378:metabolism*; D011971:Receptors, Immunologic / Q000187:drug effects; D018077:Receptors, Leukotriene; D013189:SRS-A / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012984:Software; D013844:Thiazoles / Q000494:pharmacology
chemical_list: D002867:Chromones; D003998:Dicarboxylic Acids; D007191:Indazoles; D011971:Receptors, Immunologic; D018077:Receptors, Leukotriene; D013189:SRS-A; D013844:Thiazoles; C054217:ICI 198615; C071436:cinalukast; C006312:FPL 55712; C052393:pobilukast