H 89 2HCl

Catalog No.S1582

For research use only.

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.

H 89 2HCl Chemical Structure

CAS No. 130964-39-5

Selleck's H 89 2HCl has been cited by 126 publications

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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]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
CHO  M4XUSmdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NYe3WndpOTBizszN MYGxJIg> NVv2e4xHcW6qaXLpeJMhPS2KVDDpcoR2[2WmIHPBUXAheHKxZIXjeIlwdiCmZXPy[YF{\Q>? MVe8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yOjV4OUC2PUc,OTJ3NkmwOlk9N2F-
SK-N-MC Mn7ESpVv[3Srb36gRZN{[Xl? MV:zNEDPxE1? NYPDUIpZOjRiaB?= MoHFSG1UVw>? M1fQRZJm\HWlZYOgeIhmKGSxd36tdoVofWyjdHnvckBw\iEQskGtRXIh[nliaYPvdJJwfGW{ZX7vcEBjgSB3MDZCpC=> M3HWeVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzFzN{C1OFU1Lz5zMUewOVQ2PDxxYU6=
Sf9 M{Lse2Z2dmO2aX;uJIF{e2G7 M1[5e2lvcGmkaYTpc44hd2ZiSHnzMZRi\2enZDDoeY1idiCPU1uxJIV5eHKnc4Pl[EBqdiCVZkmgZ4VtdHNuIFnDOVAhRSByLkGyJO69VS5? NWPQToY4RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMUC5PVg{PTFpPkGwPVk5OzVzPD;hQi=>
Sf9 M2ThPGZ2dmO2aX;uJIF{e2G7 NHq5XXdKdmirYnn0bY9vKG:oIILheEBTV0ONMjDlfJBz\XO|ZXSgbY4hW2Z7IHPlcIx{NCCLQ{WwJF0hOC5{NzFOwG0v NIDvRXc9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zMEm5PFM2OSd-MUC5PVg{PTF:L3G+
SF9 NHPEOmtHfW6ldHnvckBie3OjeR?= MoDkTY5pcWKrdHnvckBw\iCqdX3hckBRU0KjbIDoZUBmgHC{ZYPz[YQhcW5iU1[5JINmdGy|LDDJR|UxKD1iMj62JO69VS5? MXm8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yODl7OEO1NUc,OTB7OUizOVE9N2F-
OVCAR8 M13hdWN6fG:2b4jpZ4l1gSCjc4PhfS=> MoHyO|IhcHK| MnvPR5l1d3SxeHnjbZR6KGGpYXnud5QhcHWvYX6gU3ZESVJ6IHPlcIx{KGG|c3Xzd4VlKGG|IHfyc5d1cCCrbnjpZol1cW:wIHHmeIVzKDd{IHjyd{BjgSCPVGSgZZN{[XluIFnDOVAhRSBzMD61JO69VS5? NH\HUnY9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NEWwPFg{OCd-MkS1NFg5OzB:L3G+
OVCAR8 NVnWU5I6SW62aYDyc4xq\mW{YYTpeoUh[XO|YYm= NXSxRot7PzJiaILz NY\w[lJUSW62aYDyc4xq\mW{YYTpeoUh[WO2aY\peJkh[WejaX7zeEBpfW2jbjDPWmNCWjhiY3XscJMh[W[2ZYKgO|IhcHK|IHL5JG1VXCCjc4PhfUwhUUN3MDC9JFExNjVizszNMi=> MX:8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDN6OUWxNUc,OjR|OEm1NVE9N2F-
HL60 M1;TcmFvfGmycn;sbYZmemG2aY\lJIF{e2G7 NEH1[484OiCqcoO= M1XERmFvfGmycn;sbYZmemG2aY\lJIFkfGm4aYT5JIFo[Wmwc4SgbJVu[W5iSFy2NEBk\WyuczDh[pRmeiB5MjDodpMh[nliTWTUJIF{e2G7LDDJR|UxKD1iMUGuOEDPxE1w M1:3eFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2M{i5OVEyLz5{NEO4PVUyOTxxYU6=
PC3M M2PVVGZ2dmO2aX;uJIF{e2G7 NUDVXGZ7UW6qaXLpeIlwdiCxZjDHV2s{NWKndHGgbY4hcHWvYX6gVGM{VSClZXzsd{BjgSCHTFnTRUwhUUN3MDC9JFE2KM7:TT6= M1rFTlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF6M{S1OlA6Lz5zOEO0OVYxQTxxYU6=
U87MG MVvHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? MXXHdo94fGhiaX7obYJqfGmxbjDv[kBpfW2jbjDVPFdOTyClZXzsd{BjgSCVUlKgZZN{[XluIFnDOVAhRSBzNTFOwG0v MWG8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yQDN2NU[wPUc,OTh|NEW2NFk9N2F-
U87MG M3f0XmFvfGmycn;sbYZmemG2aY\lJIF{e2G7 M4Ls[Vk3KGi{cx?= M1n3d2FvfGmycn;sbYZmemG2aY\lJIFkfGm4aYT5JIFo[Wmwc4SgbJVu[W5iVUi3UWch[2WubIOgZYZ1\XJiOU[gbJJ{KGK7IGPSRkBie3OjeTygTWM2OCB;IEG1JO69VS5? M{i3PVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJyMUWxOlc4Lz5{MEG1NVY4PzxxYU6=
HCT116 M3Lj[2N6fG:2b4jpZ4l1gSCjc4PhfS=> MYK3NkBpenN? NYjWWYpSS3m2b4TvfIlkcXS7IHHnZYlve3RiaIXtZY4hUEOWMUG2JINmdGy|IHHzd4V{e2WmIHHzJIdzd3e2aDDpcohq[mm2aX;uJIFnfGW{IEeyJIhzeyCkeTDNWHQh[XO|YYmsJGlEPTBiPTCxOU44KM7:TT6= MXy8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDVyOEizNEc,OjR3MEi4N|A9N2F-
HCT116 MXnBcpRqeHKxbHnm[ZJifGm4ZTDhd5NigQ>? NF6yTJU4OiCqcoO= NY\ye2t2SW62aYDyc4xq\mW{YYTpeoUh[WO2aY\peJkh[WejaX7zeEBpfW2jbjDIR3QyOTZiY3XscJMh[W[2ZYKgO|IhcHK|IHL5JG1VXCCjc4PhfUwhUUN3MDC9JFE2NjdizszNMi=> MkXZQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjR|OEm1NVEoRjJ2M{i5OVEyRC:jPh?=
PC3M M{DHdGZ2dmO2aX;uJIF{e2G7 NYf6U3BbQTZiaILz NU\rNWdSUW6qaXLpeIlwdiCxZjDQR|NOKGOnbHzzJIJ6KFOUQjDhd5NigSCjZoTldkA6PiCqcoOsJGdKPTBiPTCxPEDPxE1w NXTpN4ZFRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMUe0OVEzOzVpPkG3OFUyOjN3PD;hQi=>
PC3M MoTqS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? MlvMS5Jwf3SqIHnubIljcXSrb36gc4YhcHWvYX6gVGM{VSClZXzsd{BjgSCVUlKgZZN{[XluIFnDOVAhRSBzODFOwG0v M3PUUFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF6M{S1OlA6Lz5zOEO0OVYxQTxxYU6=
LS174T NUC3bY5QTnWwY4Tpc44h[XO|YYm= NVzWRphNOjBidV2= NHLMZng3KGi{cx?= MmThTY5pcWKrdHnvckBw\iCSR1WyMZN1cW23bHH0[YQhXEOIL1zFSkB1emGwc3HjeIl3[XSrb36gbY4hcHWvYX6gUHMyPzSWIHPlcIx{KGG2IEKwJJVOKG2nYYP1doVlKGGodHXyJFYhcHK|IHL5JIR2[WxibIXjbYZmemG|ZTDy[ZBwenSncjDn[Y5mKGG|c3H5 MlXNQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjd5M{[wOlMoRjJ5N{O2NFY{RC:jPh?=
Assay
Methods Test Index PMID
Western blot p-S6K1 / p-rpS6 / p-GSK3β ; p-eNOS / T-eNOS / p-PKA ; p-GPIbβ / GPIbβ / p-BAD 26939705 31035633 29083324
Immunofluorescence myopodin ; iNOS / NFκB p65 ; p-eNOS / eNOS 17923693 30513637
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] H-89 (0.5, 1, 5 mg/kg) significantly attenuates prominent behavioral signs of morphine withdrawal in morphine-dependent mice.[6]

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 Research: [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.
  • (Only for Reference)
Animal Research:[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)
  • (Only for Reference)

Solubility (25°C)

In vitro

DMSO 104 mg/mL
(200.27 mM)
Water 6 mg/mL
(11.55 mM)
Ethanol 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.

Chemical Information

Molecular Weight 519.28
Formula

C20H20BrN3O2S.2HCl

CAS No. 130964-39-5
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles C1=CC2=C(C=CN=C2)C(=C1)S(=O)(=O)NCCNCC=CC3=CC=C(C=C3)Br.Cl.Cl

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