Bromisoval (Bromovalerylurea)

Kinase experiment:

Conditioning media are obtained from BV2 cell cultures that have been incubated for 24 h in E2 medium containing 1 μg/ml LPS, with or without Bromisoval (BU) (1-100 μg/mL) or other agents, and subjected to NO determination. To normalize the releasing NO level by the cellular protein contents, cells are solubilized with RIPA buffer (50 mM Tris-HCl, pH 8.0, 150 mM sodium chloride, 0.5% w/v sodium deoxycholate, 0.1% w/v sodium dodecyl sulfate, 1.0% w/v NP-40 substitute) and the protein contents are determined by BCA protein assay reagents[1].

Cell experiment:

Murine microglial cell line BV2 is used. BV2 cells are maintained in medium supplemented with 10% fetal bovine serum. BV2 cells are seeded onto wells in 4-well culture plates and incubated with LPS for 30 min to 24 h. When the effects of Bromisoval (BU) or other agents are investigated, BV2 cells are incubated with the appropriate agent (e.g. Bromisoval) for 30 min before the addition of LPS[1].

Animal experiment:

Suspensions of the compounds (including Bromisoval ) in aqueous 0.5% carboxymethyl cellulose are administered intraperitoneally to adult male albino mice weighing 25-35 g. All tests are performed on groups of ten mice[2].

References:

[1]. Kawasaki S, et al. Effects of hypnotic bromovalerylurea on microglial BV2 cells. J Pharmacol Sci. 2017 Jun;134(2):116-123.
[2]. Mrongovius RI, et al. Comparison of the bromureide sedative-hypnotic drugs, bromvaletone (bromisoval) and carbromal, and their chloro analogues in mice. Clin Exp Pharmacol Physiol. 1976 Sep-Oct;3(5):443-7.