H 89 2HCl

Catalog No.S1582 Batch:S158201

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Technical Data

Formula

C20H20BrN3O2S.2HCl
Molecular Weight 519.28 CAS No. 130964-39-5
Solubility (25°C)* In vitro DMSO 104 mg/mL (200.27 mM)
Water 6 mg/mL (11.55 mM)
Ethanol Insoluble
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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Biological Activity

Description H 89 2HCl is a potent PKA inhibitor with Ki of 48 nM in a cell-free assay, 10-fold selective for PKA than PKG,500-fold greater selectivity than PKC, MLCK, calmodulin kinase II and casein kinase I/II. H 89 2HCl induces autophagy.
Targets
PKA [1]
(Cell-free assay)		 S6K1 [2]
(Cell-free assay)		 PKG [1]
(Cell-free assay)
48 nM(Ki) 80 nM 0.48 μM(Ki)
In vitro

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. [1] 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. [2][3] H89 also has activity at some cellular receptors and ion channels, including Kv1.3 K+ channels,β1AR and β2AR. [4]
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. [5]

Protocol (from reference)

Kinase Assay:

[1]
  • 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:

[1]
  • Cell lines

    PC12D

  • Concentrations

    ~30 μM

  • Incubation Time

    1 h

  • Method

    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.
Animal Study:

[5] [6]
  • Animal Models

    rat; mice

  • Dosages

    20 or 200 mg/kg (Rat); 0-5 mg/kg (Mice)

  • Administration

    s.c. (Rat); i.p. (Mice)

Customer Product Validation

Data from [Data independently produced by , , J Cell Physiol, 2015, 230: 2233-2239]

Data from [Data independently produced by , , Eur J Pharm Sci, 2015, 70C: 82-91 ]

Data from [Data independently produced by , , Sci Rep, 2016, 6:26835.]

Selleck's H 89 2HCl has been cited by 179 publications

The subcortical maternal complex modulates the cell cycle during early mammalian embryogenesis via 14-3-3 [ Nat Commun, 2024, 15(1):8887] PubMed: 39406751
Proton pump inhibitors enhance macropinocytosis-mediated extracellular vesicle endocytosis by inducing membrane v-ATPase assembly [ J Extracell Vesicles, 2024, 13(4):e12426] PubMed: 38532609
Proton pump inhibitors enhance macropinocytosis-mediated extracellular vesicle endocytosis by inducing membrane v-ATPase assembly [ J Extracell Vesicles, 2024, 13(4):e12426] PubMed: 38532609
Automated, High-Throughput Phenotypic Screening and Analysis Platform to Study Pre- and Post-Implantation Morphogenesis in Stem Cell-Derived Embryo-Like Structures [ Adv Sci (Weinh), 2024, 11(4):e2304987] PubMed: 37991133
Activation of hepatic adenosine A1 receptor ameliorates MASH via inhibiting SREBPs maturation [ Cell Rep Med, 2024, 5(3):101477] PubMed: 38508143
Hepatocyte GPCR signaling regulates IRF3 to control hepatic stellate cell transdifferentiation [ Cell Commun Signal, 2024, 22(1):48] PubMed: 38233853
BMP signaling maintains auricular chondrocyte identity and prevents microtia development by inhibiting protein kinase A [ Elife, 2024, 12RP91883] PubMed: 38690987
AKAP1 alleviates VSMC phenotypic modulation and neointima formation by inhibiting Drp1-dependent mitochondrial fission [ Biomed Pharmacother, 2024, 176:116858] PubMed: 38850669
EZH2 inhibition or genetic ablation suppresses cyst growth in autosomal dominant polycystic kidney disease [ J Transl Med, 2024, 22(1):979] PubMed: 39472935
Overactive PKA signaling underlies the hyperalgesia in an ADHD mouse model [ iScience, 2024, 27(11):111110] PubMed: 39507260

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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