CHEMBL3706343

Binding Assay: Compound 2 was examined for its ability to bind human FLAP using a membrane-binding assay. The affinity of Compound 2 for human FLAP was assessed using membranes from human polymorphonuclear leukocytes and tritiated leukotriene synthesis inhibitor, 3H-3-[5-(pyrid-2-ylmethoxy)-3-tert-butylthio-1-benzyl-indol-2-yl]-2,2-dimethylpropionic acid, as a ligand. A non-limiting example of such a FLAP binding assay is as follows: Packed human polymorphonuclear cell pellets (1.8x109 cells) (Biological Speciality Corporation) were resuspended, lysed and 100,000 g membranes prepared as described (Charleson et al. Mol. Pharmacol, 41, 873-879, 1992). 100,000 g pelleted membranes were resuspended in Tris-Tween assay buffer (100 mM Tris HCl pH 7.4, 140 mM NaCl, 2 mM EDTA, 0.5 mM DTT, 5% glycerol, 0.05% Tween 20) to yield a protein concentration of 50-100 ug/mL.