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 136 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  MUTHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NF;IOIYyOCEQvF2= NFzZWWgyKGh? MkDzbY5pcWKrdIOgOU1JXCCrbnT1Z4VlKGODTWCgdJJw\HWldHnvckBl\WO{ZXHz[S=> M{XkeVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF{NU[5NFY6Lz5zMkW2PVA3QTxxYU6=
SK-N-MC NG\QOJhHfW6ldHnvckBCe3OjeR?= M2fuPVMxKM7:TR?= NEHLZ5QzPCCq MnToSG1UVw>? MUHy[YR2[2W|IITo[UBld3ewLYLl[5Vt[XSrb36gc4Yh|rJzLVHSJIJ6KGm|b4Dyc5RmemWwb3ygZpkhPTBnwrC= NX3KOmR6RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMUG3NFU1PTRpPkGxO|A2PDV2PD;hQi=>
Sf9 M2r2TWZ2dmO2aX;uJIF{e2G7 MVnJcohq[mm2aX;uJI9nKEircz30ZYdo\WRiaIXtZY4hVVONMTDlfJBz\XO|ZXSgbY4hW2Z7IHPlcIx{NCCLQ{WwJF0hOC5zMjFOwG0v MWK8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yODl7OEO1NUc,OTB7OUizOVE9N2F-
Sf9 Ml3OSpVv[3Srb36gZZN{[Xl? Mof3TY5pcWKrdHnvckBw\iC{YYSgVm9EUzJiZYjwdoV{e2WmIHnuJHNnQSClZXzsd{whUUN3MDC9JFAvOjdizszNMi=> NHnuZ4E9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zMEm5PFM2OSd-MUC5PVg{PTF:L3G+
SF9 MmfySpVv[3Srb36gZZN{[Xl? MnTQTY5pcWKrdHnvckBw\iCqdX3hckBRU0KjbIDoZUBmgHC{ZYPz[YQhcW5iU1[5JINmdGy|LDDJR|UxKD1iMj62JO69VS5? MX[8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yODl7OEO1NUc,OTB7OUizOVE9N2F-
OVCAR8 NFjlbHdEgXSxdH;4bYNqfHliYYPzZZk> M2e3cFczKGi{cx?= NWPhN4ZVS3m2b4TvfIlkcXS7IHHnZYlve3RiaIXtZY4hV1[FQWK4JINmdGy|IHHzd4V{e2WmIHHzJIdzd3e2aDDpcohq[mm2aX;uJIFnfGW{IEeyJIhzeyCkeTDNWHQh[XO|YYmsJGlEPTBiPTCxNE42KM7:TT6= NXPRc5NnRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS1NFg5OzBpPkK0OVA5QDNyPD;hQi=>
OVCAR8 MlTyRY51cXC{b3zp[oVz[XSrdnWgZZN{[Xl? NUHycZlUPzJiaILz MVrBcpRqeHKxbHnm[ZJifGm4ZTDhZ5Rqfmm2eTDh[4FqdnO2IHj1cYFvKE:YQ1HSPEBk\WyuczDh[pRmeiB5MjDodpMh[nliTWTUJIF{e2G7LDDJR|UxKD1iMUCuOUDPxE1w NEfKR|U9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NEO4PVUyOSd-MkSzPFk2OTF:L3G+
HL60 M4W3WmFvfGmycn;sbYZmemG2aY\lJIF{e2G7 NEG1UWk4OiCqcoO= NIDiNpZCdnSrcILvcIln\XKjdHn2[UBi[3Srdnn0fUBi\2GrboP0JIh2dWGwIFjMOlAh[2WubIOgZYZ1\XJiN{KgbJJ{KGK7IF3UWEBie3OjeTygTWM2OCB;IEGxMlQh|ryPLh?= M1znflxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2M{i5OVEyLz5{NEO4PVUyOTxxYU6=
PC3M NFfqdHVHfW6ldHnvckBie3OjeR?= NH7EUmpKdmirYnn0bY9vKG:oIFfTT|Mu[mW2YTDpckBpfW2jbjDQR|NOKGOnbHzzJIJ6KEWOSWPBMEBKSzVyIE2gNVUh|ryPLh?= NX;Lbm9NRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMUizOFU3ODlpPkG4N|Q2PjB7PD;hQi=>
U87MG NUHhXWtZT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= M4jCVmdzd3e2aDDpcohq[mm2aX;uJI9nKGi3bXHuJHU5P02JIHPlcIx{KGK7IGPSRkBie3OjeTygTWM2OCB;IEG1JO69VS5? MWm8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yQDN2NU[wPUc,OTh|NEW2NFk9N2F-
U87MG NFHWdWZCdnSrcILvcIln\XKjdHn2[UBie3OjeR?= MXe5OkBpenN? NYPYVHZlSW62aYDyc4xq\mW{YYTpeoUh[WO2aY\peJkh[WejaX7zeEBpfW2jbjDVPFdOTyClZXzsd{Bi\nSncjC5OkBpenNiYomgV3JDKGG|c3H5MEBKSzVyIE2gNVUh|ryPLh?= NX3ve3MxRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkCxOVE3PzdpPkKwNVUyPjd5PD;hQi=>
HCT116 MXLDfZRwfG:6aXPpeJkh[XO|YYm= NVW1eYJtPzJiaILz NHjE[VdEgXSxdH;4bYNqfHliYXfhbY5{fCCqdX3hckBJS1RzMU[gZ4VtdHNiYYPz[ZN{\WRiYYOg[5Jwf3SqIHnubIljcXSrb36gZYZ1\XJiN{KgbJJ{KGK7IF3UWEBie3OjeTygTWM2OCB;IEG1Mlch|ryPLh?= NV;wfJV{RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS1NFg5OzBpPkK0OVA5QDNyPD;hQi=>
HCT116 Mmr4RY51cXC{b3zp[oVz[XSrdnWgZZN{[Xl? Mn7DO|IhcHK| NFXacZNCdnSrcILvcIln\XKjdHn2[UBi[3Srdnn0fUBi\2GrboP0JIh2dWGwIFjDWFEyPiClZXzsd{Bi\nSncjC3NkBpenNiYomgUXRVKGG|c3H5MEBKSzVyIE2gNVUvPyEQvF2u NF3PV409[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NEO4PVUyOSd-MkSzPFk2OTF:L3G+
PC3M NEK0OYZHfW6ldHnvckBie3OjeR?= MX[5OkBpenN? NFnlfHBKdmirYnn0bY9vKG:oIGDDN20h[2WubIOgZpkhW1KEIHHzd4F6KGGodHXyJFk3KGi{czygS2k2OCB;IEG4JO69VS5? MkjaQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTd2NUGyN|UoRjF5NEWxNlM2RC:jPh?=
PC3M NHrK[pBIem:5dHigbY5pcWKrdHnvckBie3OjeR?= M{DrS2dzd3e2aDDpcohq[mm2aX;uJI9nKGi3bXHuJHBEO01iY3XscJMh[nliU2LCJIF{e2G7LDDJR|UxKD1iMUig{txONg>? M1fuUVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF6M{S1OlA6Lz5zOEO0OVYxQTxxYU6=
LS174T MlH0SpVv[3Srb36gZZN{[Xl? NYrTUYhiOjBidV2= NWW1[ldZPiCqcoO= M{nnXmlvcGmkaYTpc44hd2ZiUFfFNk1{fGmvdXzheIVlKFSFRj;MSWYhfHKjboPhZ5RqfmG2aX;uJIlvKGi3bXHuJGxUOTd2VDDj[YxteyCjdDCyNEB2VSCvZXHzeZJm\CCjZoTldkA3KGi{czDifUBlfWGuIHz1Z4ln\XKjc3WgdoVxd3K2ZYKg[4Vv\SCjc4PhfS=> NELxN409[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{N{ezOlA3Oyd-Mke3N|YxPjN:L3G+
Assay
Methods Test Index PMID
Western blot p-eNOS / T-eNOS / p-PKA ; p-S6K1 / p-rpS6 / p-GSK3β ; p-GPIbβ / GPIbβ / p-BAD 31035633 26939705 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.
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)

Solubility (25°C)

In vitro

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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