Cell lines

U2OS cells

Preparation method

The solubility of this compound in DMSO is >18.6 mg/mL. General tips for obtaining a higher concentration: Please warm the tube at 37 ℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below - 20 ℃ for several months.

Reacting condition

1 μM; 1 hr

Applications

In the U2OS cells transfected with a GFP-BAZ2A fusion construct, GSK2801 reduced FRAP half-recovery time to the same extent as observed for the U2OS cells transfected with a mutant construct after SAHA treatment, which indicated that GSK2801 displaced BAZ2A from chromatin.

Animal models

Male CD1 mice

Dosage form

30 mg/kg; i.p. or p.o.

Applications

In a male CD1 mouse, a single dose of GSK2801 (30 mg/kg; p.o.) resulted in reasonable in vivo exposure and plasma stability, as well as modest clearance (Tmax = 1.0 hr; Cmax = 435 ng/mL; T1/2 = 1.5 hrs), which implied that GSK2801 could be used as BAZ2A/B bromodomain inhibitor in vivo.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

GSK2801, as an inhibitor of BAZ2A and BAZ2B bromodomains, is potent, selective and cell active acetyl-lysine competitive, with dissociation constants (KD) of 136 and 257 nM for binding to BAZ2B and BAZ2A, respectively [1].

Bromodomains, as protein interaction domains, are acetyl-lysine specific. Bromodomain containing proteins BAZ2A and BAZ2B are closely related. The nucleolar remodeling complex (NoRC) regulates the expression of noncoding RNAs. BAZ2A and BAZ2B constitute the central scaffolding protein of NoRC [1].

In U2OS cells, treatment with the SAHA induced hyperacetylated chromatin. In a GFP-BAZ2A fusion construct, the conserved asparagines that are essential for recognizing the acetylated lysine has been mutated. When a GFP-BAZ2A fusion construct was transfected into SAHA-treated U2OS cells, the mutant construct accelerated FRAP half-recovery time. Treatment with GSK2801 alone in U2OS cells also accelerated FRAP half-recovery time. Both acceleration extents are the same. This meant that GSK2801 can displace BAZ2A from chromatin [1].

Pharmacokinetic parameters of GSK2801 after intraperitoneal and oral dosing to male CD1 mice were measured. Data showed that after oral dosing in vivo, GSK2801 has reasonable exposure, reasonable plasma stability and modest clearance. GSK2801 can be used as an inhibitor of BAZ2A/B bromodomain in vivo [1]. No more in vivo data of the treatment with GSK2801 are found.

Reference:
[1].  Chen P, Chaikuad A, Bamborough P, et al. Discovery and Characterization of GSK2801, a Selective Chemical Probe for the Bromodomains BAZ2A and BAZ2B. Journal of medicinal chemistry, 2015.