GW 791343 HCl

483, 81

0 uM -10 uM

In cells expressing human P2X7 receptors, GW 791343 inhibits agonist-stimulated ethidium accumulation in both sucrose and NaCl buffer. In NaCl buffer, GW 791343 reduces the maximal response to both ATP and BzATP, but there is little effect on agonist potency except for a decrease in the presence of 300–1000 nM GW 791343. GW 791343 also reduces maximal responses to ATP and BzATP in sucrose buffer, although this effect is more marked when using ATP as agonist. In sucrose buffer, GW 791343 produces a slight decrease in ATP potency at 300 nM. GW 791343 decreases BzATP potency at concentrations of 300 nM to 10 uM. A more marked increase in agonist effect is observed when using ATP as agonist in NaCl buffer with GW791343 increasing the pEC50 and maximal response to ATP at concentrations of 10 and 30 uM. In sucrose buffer, GW791343 also increases responses when using ATP as agonist. [1] GW791343 inhibits responses at the human–rat chimeric receptor in both sucrose and NaCl buffer. GW791343 increases responses tATP at the F95L mutant receptor. [2] GW791343 is a non-competitive antagonist and negative allosteric modulator at the human P2X7 receptor, however, GW 791343 also acts as a positive allosteric modulator at the rat P2X7 receptor. [3] At the dog P2X7 receptor, GW 791343 is an antagonist with similar potency to that determined at the human receptor. [4]

HEK293 cells, expressing rat or human recombinant P2X7 receptors, are grown (18–24 hours) to form a completely confluent monolayer in poly-L-lysine pretreated 96-well plates. Studies are performed using assay buffers comprising (in mM): 10 mM HEPES, 5 mM N-methyl-D-glucamine, 5.6 mM KCl, 10 mM D-glucose, 0.5 mM CaCl2 (pH 7.4) and supplemented with either 280 mM sucrose (sucrose buffer) or 140 mM NaCl (NaCl buffer). Before use, growth media is completely removed from the cells and they are rinsed with 350 uL of the appropriate assay buffer, which is also removed before performing assay additions. All solutions are aspirated using 25-gauge bevelled syringe needles to provide complete solution removal. In all studies, the final assay volume is 100 uL. The majority of studies are performed at room temperature (19–21C). In one study, the effect of GW 791343 at rat P2X7 receptors is studied at 4 C in sucrose buffer.

2-(3, 4-difluorophenylamino)-N-(2-methyl-5-(piperazin-1-ylmethyl)phenyl)acetamide trihydrochloride