title: Binding of a novel radioiodinated thromboxane A2/prostaglandin H2 antagonist to guinea pig lung membranes.
abstract: The utilization of a novel radioiodinated TXA2/PGH2 receptor antagonist, ISAP (7-[(1R,2S,3S,5R)-6,6-dimethyl-3(4- iodobenzenesulfonylamino)-bicyclo[3.1.1]-hept-2-yl]-5(Z)-heptenoic acid) to characterize TXA2/PGH2 receptors from guinea pig lung parenchymal membranes in radioligand binding assays is described. [125I]ISAP binding was saturable, displaceable, and dependent upon protein concentration. The time course of binding yielded k1 = 2.12 x 10(8) M-1 min-1, k1 = 4.46 x 10(-3) min-1, Kd = k-1/k1 = 17.8 pM. Equilibrium binding studies indicated a single class of high affinity binding sites with a Kd of 52.7 +/- 1.9 pM and a Bmax of 92.7 +/- 7.2 fmoles/mg protein (n = 4). Binding was inhibited by a series of structurally diverse mimetics and antagonists with the rank order of potency IBOP greater than ONO11113 = SQ26655 greater than U46619 (mimetics) and (d)-S-145 greater than ISAP greater than (1)-S-145 greater than SQ29548 greater than BM13505 = I-PTA-OH (antagonists), with entantioselectivity of binding demonstrated by (d) and (1) S-145. Binding was also inhibited by prostanoids (PGD2, PGF2 alpha, and 9 alpha, 11 beta-PGF2) thought to act at the airway TXA2/PHH2 receptor, but not by histamine or carbachol, and only weakly by LTB4 and LTD4, consistent with specific binding to the lung TXA2/PGH2 receptor.
authors.affiliation: Department of Pulmonary Research Lilly Research Laboratories, Lilly Corporate Center, Indianapolis, IN 46285.
authors.forename: D L
authors.initials: DL
authors.lastname: Saussy
authors.forename: P D
authors.initials: PD
authors.lastname: Clark
authors.forename: D L
authors.initials: DL
authors.lastname: Gunn
authors.forename: D E
authors.initials: DE
authors.lastname: Mais
authors.forename: L L
authors.initials: LL
authors.lastname: Froelich
mesh_terms: D000818:Animals; D001643:Bridged Bicyclo Compounds / Q000378:metabolism*; D005229:Fatty Acids, Monounsaturated / Q000378:metabolism*; D006168:Guinea Pigs; D007457:Iodine Radioisotopes; D008168:Lung / Q000378:metabolism*; D008566:Membranes / Q000378:metabolism; D015394:Molecular Structure; D044262:Prostaglandin H2; D011463:Prostaglandins H / Q000037:antagonists & inhibitors*; D011869:Radioligand Assay; D017482:Receptors, Thromboxane / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D012995:Solubility; D013928:Thromboxane A2 / Q000037:antagonists & inhibitors*
chemical_list: D001643:Bridged Bicyclo Compounds; D005229:Fatty Acids, Monounsaturated; D007457:Iodine Radioisotopes; D011463:Prostaglandins H; D017482:Receptors, Thromboxane; C076030:7-(6,6-dimethyl-3-(4-iodobenzenesulfonylamino)bicyclo(3.1.1.)hept-2-yl)-5-heptenoic acid; D044262:Prostaglandin H2; D013928:Thromboxane A2