CHARACTERIZATION AND AUTORADIOGRAPHIC MAPPING OF [H-3] CP96,345, A NONPEPTIDE SELECTIVE NK1 RECEPTOR ANTAGONIST IN GUINEA-PIG LUNG

We have studied binding and distribution of NK1 receptors in guinea pig lung using [H-3]CP96,345. Kinetic studies showed that specific binding of [H-3]CP96,345 was rapid and reversible, giving a kinetic dissociation constant (K-d) of 0.28 +/- 0.05 nM. The specific binding was also saturable and Scatchard analysis indicated a single class of binding site with an equilibrium K-d of 0.12 +/- 0.03 nM and maximum binding capacity (B-max) of 107.0 +/- 10.3 fmol/mg of protein. Competition studies showed the rank order of affinity for agonists and antagonists as follows: SP > NKA = septide much greater than NKB = senktide; CP96,345 > FK888 > FK224 > L668169. NK3 agonists, NK2-selective antagonists, and a calcium channel blocker, diltiazem, showed no displacement, indicating high selectivity for NK1 receptors. Autoradiographic mapping showed specific labeling over airway smooth muscle from central to peripheral airways, submucosal glands, and nerve fibers of trachea. The labeling of airway epithelium was increased with diminishing size of airways. Pulmonary blood vessels were also moderately labeled and there was sparse labeling over alveolar walls. [H-3]CP96,345 may provide a useful tool to evaluate NK1 receptor expression in peripheral organs.