Cell lines

PBMC cells

Preparation method

The solubility of this compound in DMSO is >10 mM. 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.

Reaction Conditions

100 nM, 24 hours for hyperacetylation

Applications

To test the acetylation ability of ITF2357, rested PBMCs were preincubated with the inhibitor for 1 h at 37° C and then stimulated with LPS. After 3, 6, and 24 h, extracts of the cell pellets were made and the acetylated lysines were determined in total cellular extracts. After 3 h of incubation with LPS in the presence of ITF2357, hyperacetylation is clearly present. However, a greater duration of hyperacetylation up to 24 h was observed in cells exposed to ITF2357.

Animal models

BALB/C and C57BL/6 mice

Dosage form

Gavage, 5 mg/kg (BALB/C mice) Gavage, 1 or 10 mg/kg (C57BL/6 mice)

Applications

Mice were given 100 μL water or ITF2357 (5 mg/kg) by gavage and, after 1 h, injected intravenously with 200 μg/mouse of ConA. Mice were bled 24 h later for evaluation of serum ALT levels. ITF2357 pretreatment reduced more than 80% of the ALT levels. A dose of 1 mg/kg ITF2357 was as effective as a dose of 10 mg/kg in reducing ConA hepatitis as measured by ALT levels.

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.

ITF2357, also known as givinostat, is a potent inhibitor of both class I and class II histone deacetylase (HDAC) as well as a potent inhibitor of hematopoietic colony formation by JAKE^V617F-bearing progenitor cells from chronic myeloproliferative neoplasmsin vitro. Previous studies has shown that ITF2357 induces apoptosis of multiple myeloma (MM) and acute myelogenous leukemia (AML) cells following induction of p21 and down-modulation of Bcl-2 and Mcl-1 proteins and inhibits the production of pro-inflammatory cytokines (such as IL-1, IL-6, tumor necrosis factor (TNF)-α and interferon (IFN)-γ) by peripheral blood mononuclear cells as well as the production of IL-6 and vascular endothelium growth factor (VEGF) by mesenchymal stromal cells.

Reference

[1].Katia Todoerti, Valentina Barbui, Olga Pedrini, Marta Linett, Gianluca Fossati, Paolo Mascagni, Alessandro Rambaldi, Antonino Neri, Martino Introna, Luigia Lombardi, and Josee Golay. Pleiotropi anti-myeloma activity of ITF2357: inhibition of interleukin-6 receptor signaling and repression of miR-19a and miR-19b. Haematologica 2010; 95(2): 260-269