Male Wistar rats (150-250 g) were dosed ip with compound 57 at 2-20mk/kg

Membranes from Chinese hamster ovary CHO cells expressing human recombinant 5-HT2A receptors were used for activity evaluation. The binding studies were performed in 96-well plates (Dynatech, Billinghurst, UK) in a total volume of 250 mkl, consisting of the radioligand [3H] Ketanserin (0.5 nM), drug (P-88-8991) and CHO membrane preparation diluted in 50 mM Tris buffer with pH 7.7. The plates were incubated for 15 min at 22 C Nonspecific binding was determined in the presence of ketanserin (10 mkM). Reactions were terminated by flash filtration and inverse transfer to 96-well filter plates [Perkin Elmer/Packard Bioscience, Mericlen, CT, USA 96-well cell harvester, filter plates GFC, coated with plasminogen activator inhibitor (PAI) as necessary]. The plates were dried for 30 min at 56 C and sealed at the bottom with an adhesive sheet (Topseal; Perkin Elmer/Packard Bioscience). Subsequently, 50 mkl of scintillation fluid (Microscint-20; Perkin Elmer/Packard Bioscience) was added to each well, the plates sealed on top and the radioactivity counted in a 96-well plate counter (Topcount; Perkin Elmer/Packard Bioscience).