COMPARISON OF TACHYKININ NK(1) RECEPTORS IN HUMAN IM9 AND U373-MG CELLS, USING ANTAGONIST (FK888, (PLUS-OR-MINUS)-CP-96,345, AND RP-67580) BINDING

We have used one peptide (FK888) and two non-peptide ((+/-)-CP-96,345 and RP 67580) antagonists, along with the preferred endogenous agonist, substance P, to compare the pharmacological (binding) profile of NK1 receptors expressed by human B lymphoblastoma (IM9) and astrocytoma (U373 MG) cells. Of the ligands tested, substance P was the most potent in both cell lines: binding affinities were 0.1 nM for IM9 cells, and 0.3 nM for U373 MG cells, respectively. The high-affinity dipeptide antagonist, FK888, bound to NK1 receptors in both cell lines with similar potencies: K(i) values were 1.2 nM and 3.6 nM for IM9 cells and U373 MG cells, respectively. Of the non-peptide antagonists, as expected, (+/-)-CP-96,345 displayed higher affinity (0.4 nM in IM9 cells, and 1.2 nM in U373 MG cells) than did RP 67580 (33 nM and 223 nM in IM9 cells and U373 MG cells, respectively) in both cell lines. We conclude that the pharmacological profile of NK1 receptors is similar in the human lymphoblastoma and astrocytoma cells, i.e. if NK1 receptor subtypes exist in humans, these cell lines are likely to express a similar subtype. Because IM9 cells grow faster and are easier to maintain, this cell line may be preferable to the astrocytoma cells as a primary screen to identify NK1 receptor antagonists.