AZD 9272 

Saturable binding and competition binding studies utilize incubations of 1 hour at 22°C. For saturation studies, membranes from mGluR5-GHEK cells are incubated with increasing concentrations (0.1 to 30 nM) of [³H]AZD9272, in the presence or absence of 10 μM MPEP. In a variation of these studies, saturable [³H]AZD9272 binding is determined in the presence of low concentrations (10 and 20 nM) of MPEP. Consistency of the B_max in the presence or absence of MPEP supports the interaction of these ligands with a unitary binding site^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

hmGluR5-GHEK cells are seeded onto 96 well plates at 50,000 cells/well in media containing 10 µCi/mL [³H]myo-inositol. Cells are incubated overnight (16 h), then washed three times and incubated for 1 hour at 37°C in HEPES buffered saline supplemented with 1 unit/mL glutamate pyruvate transaminase and 2 mM pyruvate. Cells are washed once in HEPES buffered saline and pre-incubated for 10 minutes in HEPES buffered saline containing 10 mM LiCl. Antagonist activity is determined by pre-incubating cells with AZD9272 for 10 minutes, then incubating for 30 minutes at 37°C in the presence of glutamate (EC₈₀, 80 µM). AZD9272 is tested at 10 concentrations between 1 nM and 30 μM, in duplicate. The reaction is terminated by the addition of 0.1 mL perchloric acid (5%) on ice, with incubation at 4°C for at least 30 minutes^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Approximately 48 male Wistar rats weighing 240 to 250 g at the beginning of the experiments are housed in pairs, or group housed up to 8 rats per cage, in a colony room with water accessible at all times and lights on between 6:00 AM and 6:00 PM; by restricting access to food, animals are kept at approximately 80% of free feeding weight. All animals are divided into different groups and trained to discriminate cocaine (3.4 mg/kg i.p., 15 minutes), PCP (1.6 mg/kg i.p., 30 minutes), MTEP (2 mg/kg i.p., 30 minutes), or AZD9272 (1.6 mg/kg p.o., 60 minutes) from no drug^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Swedberg MD, et al. AZD9272 and AZD2066: selective and highly central nervous system penetrant mGluR5 antagonists characterized by their discriminative effects. J Pharmacol Exp Ther. 2014 Aug;350(2):212-22.  [Content Brief]

  [2]. Raboisson P, et al. Discovery and characterization of AZD9272 and AZD6538-Two novel mGluR5 negative allosteric modulators selected for clinical development. Bioorg Med Chem Lett. 2012 Nov 15;22(22):6974-9.  [Content Brief]