title: Characterization of the pharmacological profile of the potent LTB4 antagonist CP-105,696 on murine LTB4 receptors in vitro.
abstract: 1. Binding of [3H]-leukotriene B4 ([3H]-LTB4) to murine spleen membranes (MSM) was determined. 2. Scatchard analyses of [3H]-LTB4 binding indicated the presence of high (KD1 = 1.7 nM) and low (KD2 = 7.5 nM) affinity receptors on MSM with Bmax values of 151 fmol mg-1 protein (Bmax1) and 354 fmol mg-1 protein (Bmax2), respectively. 3. CP-105,696, a potent LTB4 antagonist, inhibited [3H]-LTB4 (0.67 nM) binding to the high affinity receptor on MSM, IC50 = 30.2 nM, Ki = 17.7 nM with a Hill coefficient of 0.93. 4. Scatchard analyses of [3H]-LTB4 binding to MSM in the presence of CP-105,696 indicated that the high-affinity receptor was inhibited in a non-competitive manner and the low-affinity receptor in a competitive manner. 5. Isolated peripheral blood murine neutrophils (MN) responded chemotactically to LTB4, EC50 = 2.5 nM. CP-105,696 blocked this response, IC50 = 2.3 nM. When examined over a full concentration-response range of LTB4, CP-105,696 inhibited chemotaxis in a non-competitive manner. 6. Murine neutrophils in anticoagulated whole blood upregulated the integrin, complement receptor type 3 (CD11b/CD18, Mac-1) in response to LTB4, EC50 = 20 nM and this was inhibited by CP-105,696 in a competitive manner. 7. These results provide evidence that MSM have specific binding sites for LTB4, and as exemplified by CP-105,696, that these receptors may be useful for determining the potency and nature of antagonism of novel LTB4 receptor antagonists.
authors.affiliation: Department of Cancer, Pfizer Inc., Groton, CT 06340, USA.
authors.forename: H J
authors.initials: HJ
authors.lastname: Showell
authors.forename: R
authors.initials: R
authors.lastname: Breslow
authors.forename: M J
authors.initials: MJ
authors.lastname: Conklyn
authors.forename: G P
authors.initials: GP
authors.lastname: Hingorani
authors.forename: K
authors.initials: K
authors.lastname: Koch
mesh_terms: D000818:Animals; D001578:Benzopyrans / Q000378:metabolism / Q000494:pharmacology*; D001665:Binding Sites; D001667:Binding, Competitive; D002264:Carboxylic Acids / Q000378:metabolism / Q000494:pharmacology*; D002634:Chemotaxis, Leukocyte; D004305:Dose-Response Relationship, Drug; D007975:Leukotriene B4 / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000494:pharmacology*; D016177:Macrophage-1 Antigen / Q000378:metabolism; D051379:Mice; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism; D018102:Receptors, Leukotriene B4 / Q000037:antagonists & inhibitors* / Q000187:drug effects / Q000378:metabolism*; D013154:Spleen / Q000378:metabolism
chemical_list: D001578:Benzopyrans; D002264:Carboxylic Acids; D016177:Macrophage-1 Antigen; D018102:Receptors, Leukotriene B4; D007975:Leukotriene B4; C089698:CP 105696