product name H 89 2HCl
Description: H 89 2HCl is a potent PKA inhibitor with Ki of 48 nM in a cell-free assay, it is 10-fold selective for PKA than PKG, and showed 500-fold greater selectivity than PKC, MLCK, calmodulin kinase II and casein kinase I/II. In PC12D cells, pretreatment with H-89 dose-dependently inhibited the forskolin-induced protein phosphorylation, with no influence in intracellular cyclic AMP levels. In the hypotonic medium, 50 μM H89, a concentration commonly used to inhibit PKA, prevented the redistribution response. In normal medium, H89 (50 Μm) induced the redistribution of ERGIC 53 to the ER by 20 min.
References: J Biol Chem. 1990 Mar 25;265(9):5267-72; Cardiovasc Drug Rev. 2006 Fall-Winter;24(3-4):261-74.
519.28
Formula
C20H20BrN3O2S.2HCl
CAS No.
130964-39-5
Storage
-20℃ for 3 years in powder form
-80℃ for 2 years in solvent
Solubility (In vitro)
DMSO: 104 mg/mL (200.3 mM)
Water: 6 mg/mL (11.6 mM)
Ethanol: <1 mg/mL
Solubility (In vivo)
1% DMSO+30% polyethylene glycol+1% Tween 80: 30mg/mL
Synonyms
other peoduct :
| In Vitro |
In vitro activity: H89 2HCl is a potent PKA (cAMP-dependent) protein kinase A inhibitor with Ki of 48 nM, exhibits 10-fold selectivity over PKG, exhibits >500-fold selectivity over PKC, MLCK, calmodulin kinase II and casein kinase I/II. Pretreatment of the cells with H-89 (30 μM) 1 h before the addition of forskolin markedly inhibits the forskolin-induced protein phosphorylation in a dose-dependent manner. H89 also inhibits several other kinases with IC50 of 80, 120, 135, 270, 2600 and 2800 nM for S6K1, MSK1, PKA, ROCKII, PKBα and MAPKAP-K1b, respectively. H89 also has activity at some cellular receptors and ion channels, including Kv1.3 K+ channels,β1AR and β2AR. Kinase Assay: PKA enzyme activity: cAMP-dependent protein kinase activity is assayed in a reaction mixture containing, in a final volume of 0.2 mL, 50 mM Tris-HC1 (pH 7.0), 10 mM magnesium acetate, 2 mM EGTA, 1 μM cAMP or absence of cAMP, 3.3-20 μM [γ-32P]ATP (4 × 105 cpm), 0.5 μg of the enzyme, 100 μg of histone H2B, and each compound, as indicated. Cell Assay: Levels of intracellular cAMP are determined. After 48 h in culture, PC12D cells are cultured in test medium containing 30 μM H-89 for 1 h and then exposed to fresh medium that contained both 10 μM forskolin and 30 μM H-89. Cells are scraped off with a rubber policeman and sonicated in the presence of 0.5 ml of 6% trichloroacetic acid. To extract trichloroacetic acid, 2 ml of petroleum ether is added, the preparation mixed and centrifuged at 3000 rpm for 10 min. After aspiration of the upper layer, the residue sample solution is used for determination. |
|---|---|
| In Vivo | H89 causes distinct modifications of protein phosphorylation, with the most robust changes in phosphorylation are fructose-1,6-biphosphatase, heterogeneous nuclear ribonucleoprotein (hnRNP), NSFL1 cofactor p47, all which have potentially regulatory connections to cAMP/PKA. H-89 (0.5, 1, 5 mg/kg) significantly attenuates prominent behavioral signs of morphine withdrawal in morphine-dependent mice. |
| Animal model | Mouse and rat |
| Formulation & Dosage | Dissolved in 100% DMSO, diluted 1:20 in 0.9% sterile saline (Rat); 1% DMSO (Mice); 20 or 200 mg/kg (Rat); 0-5 mg/kg (Mice); s.c. (rat); i.p. (mice) |
| References | J Biol Chem. 1990 Mar 25;265(9):5267-72; Cardiovasc Drug Rev. 2006 Fall-Winter;24(3-4):261-74; J Pharmacol Exp Ther. 2006 Aug;318(2):589-95; Sci Signal. 2008 Jun 3;1(22):re4. |
