Home TGF-beta/Smad PKC inhibitor Bisindolylmaleimide IX (Ro 31-8220) Mesylate For research use only.

Bisindolylmaleimide IX (Ro 31-8220) Mesylate

Bisindolylmaleimide IX (Ro 31-8220 Mesylate) is a pan-PKC inhibitor with IC50 of 5 nM, 24 nM, 14 nM, 27 nM, and 24 nM for PKC-α, PKC-βI, PKC-βII, PKC-γ, and PKC-ε, respectively, and also shows potent inhibition against MAPKAP-K1b, MSK1, GSK3β and S6K1.

Bisindolylmaleimide IX (Ro 31-8220) Mesylate Chemical Structure

Bisindolylmaleimide IX (Ro 31-8220) Mesylate Chemical Structure

CAS No. 138489-18-6

Purity & Quality Control

Bisindolylmaleimide IX (Ro 31-8220) Mesylate Related Products

Signaling Pathway

PKC Signaling Pathways

PKC Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
Sf9 Function assay Inhibition of His-tagged human MSK1 expressed in Sf9 cells, IC50=0.008μM 10998351
Sf21 Function assay Inhibition of His-tagged human GSK3b expressed in Sf21 cells, IC50=0.038μM 10998351
HCT116 Antiproliferative assay 48 hrs Antiproliferative activity against human HCT116 cells over expressing RSK2 after 48 hrs by MTT assay, IC50=0.84μM 21488662
PC3 Cytotoxicity assay 48 hrs Cytotoxicity against human PC3 cells assessed as growth inhibition after 48 hrs by MTT assay, IC50=1.74μM 23434140
MDA-MB-231 Antiproliferative assay 48 hrs Antiproliferative activity against human MDA-MB-231 cells over expressing RSK2 after 48 hrs by MTT assay, IC50=1.77μM 21488662
MCF7 Antiproliferative assay 48 hrs Antiproliferative activity against human MCF7 cells over expressing RSK2 after 48 hrs by MTT assay, IC50=1.96μM 21488662
MCF7 Cytotoxicity assay 48 hrs Cytotoxicity against human MCF7 cells assessed as growth inhibition after 48 hrs by MTT assay, IC50=1.96μM 23434140
DE3 Function assay Inhibition of human full length SIRT1 expressed in DE3 cells by fluorimetric assay, IC50=25μM 19734050
A549 Function assay Inhibition of SIRT2 in A549 cells assessed as ability to induce hyperacetylation of tubulin by Western blot analysis 17149860
A549 Function assay 10 uM Inhibition of SIRT2 in A549 cells assessed as ability to induce hyperacetylation of tubulin at 10 uM by Western blot analysis 17149860
MCF7 Function assay 10 uM 16 hrs Inhibition of SIRT2 in human MCF7 cells assessed as tubulin hyperacetylation at 10 uM after 16 hrs by Western blot 20030343
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Biological Activity

Description Bisindolylmaleimide IX (Ro 31-8220 Mesylate) is a pan-PKC inhibitor with IC50 of 5 nM, 24 nM, 14 nM, 27 nM, and 24 nM for PKC-α, PKC-βI, PKC-βII, PKC-γ, and PKC-ε, respectively, and also shows potent inhibition against MAPKAP-K1b, MSK1, GSK3β and S6K1.
Targets
PKCα [1]
(Cell-free assay)		 PKCβ2 [1]
(Cell-free assay)		 PKCβ1 [1]
(Cell-free assay)		 PKCε [1]
(Cell-free assay)		 PKCγ [1]
(Cell-free assay)
5 nM 14 nM 24 nM 24 nM 27 nM
In vitro
In vitro Ro 31-8220 inhibits rat brain PKC activity with IC50 of 23 nM, and does not show any high degree of selectivity between PKC-α, PKC-β, PKC-γ, and PKC-ε. [1] Ro 31-8220 also inhibits MSK1, MAPKAPK1, RSK, GSK3β and S6K1 with a potency similar to that for PKC. In addition, Ro 31-8220 inhibits voltage-dependent Na+ channels. [2] Ro 31-8220 alters cellular protein kinase C localization and potently inhibits growth of A549 and MCF-7 cells with IC50 of 0.78 μM and 0.897 μM, respectively. [3] RO 31-8220 enhances epinephrine-induced platelet aggregation in catecholamine hypo-responsive platelets by enhancing Akt phosphorylation. [4] Ro 31-8220 significantly decreases apoE secretion from primary human macrophages by inhibiting vesicular transport of apoE to the plasma membrane without significantly affecting apoE mRNA or apoE protein levels. [5]
Kinase Assay Assay of PKC Activity
Assay mixtures contain 0.2 mg/mL peptide-gamma, 10 μM MgCl2, 0.6 mM CaCl2, 10 μM [γ-32P]ATP, 1.25 mg/mL phosphatidylserine and 1.25 ng/mL phorbol 12-myristate 13-acetate in 20 mM Hepes (pH 7.5), 2 mM EDTA, 1 mM dithiothreitol and 0.02% (w/v) Triton X-100. Peptide-γ is a synthetic peptide, GPRPLFCRKGSLRQKW, resembling the PKC-γ pseudosubstrate site, except that a serine residue replaces the pseudosubstrate alanine, converting the peptide from an inhibitor into a substrat. The assays are started by the addition of 2.5 m-units of enzyme, incubated at 30 °C for 10 min and terminated by spotting on to P81 paper, followed by extensive washing in 75 mM orthophosphoric acid. The papers are then washed in ethanol, dried, and incorporated radioactivity is determined by liquidscintillation spectroscopy.
Cell Research Cell lines Human A549 lung and MCF-7 breast carcinoma cells
Concentrations 2.5 μM
Incubation Time 24-48 hours
Method

Human A549 lung and MCF-7 breast carcinoma cells are obtained from the European Collection of Animal Cell Cultures. Cells (passage number 10-30) are cultured in an atmosphere of 5% carbon dioxide, the former in Ham's F-12 medium with penicillin/streptomycin, the latter in minimum essential medium (Eagle's modification) with additional pyruvate (1 mM) and non-essential amino acids. Both media are supplemented with 10% FCS and glutamine (2 mM). Cells are subcultured routinely twice weekly to maintain logarithmic growth. For cell proliferation studies cells are seeded and incubated with 3 ml of medium including agents, which is replenished at intervals of 48 h (A549) or 72 h (MCF-7). Following incubation for 4 days (A549) or 6 days (MCF-7) with drugs, cell number is assessed using a Coulter Counter Model ZM. In order to achieve PKC depletion, cells are incubated for 24 h with bryostatin 1 (1 μM). Under these conditions growth inhibition caused by bryostatin 1 is negligible. Bryostatin is removed by extensive washing of the cells followed by a 2 h recovery period. In previous work using the A549 cell line this washing procedure has been shown to eliminate bryostatin-mediated effects. The cells are then incubated for a further 24 h with staurosporine, RO 31-8220, UCN-01 or H-7. In some experiments cells are incubated with inhibitor for 48 rather than 24 h, in this case bryostation was not removed and left in the incubate. After removal of agents inhibition of DNA synthesis is evaluated by measurement of [3H]Tdr incorporation into cell. Radioactivity is counted using a Packard 1500 Tricarb scintillation counter.
In Vivo
In vivo In MLP−/− Mice, Ro 31-8220 (6 mg/kg/d, s.c.) results in a significant increase in cardiac contractility. [6]
Animal Research Animal Models MLP−/− Mice
Dosages 6 mg/kg/d
Administration s.c.

Chemical Information & Solubility

Molecular Weight 553.65 Formula

C25H23N5O2S.CH4O3S
CAS No. 138489-18-6 SDF Download Bisindolylmaleimide IX (Ro 31-8220) Mesylate SDF
Smiles CN1C=C(C2=CC=CC=C21)C3=C(C(=O)NC3=O)C4=CN(C5=CC=CC=C54)CCCSC(=N)N.CS(=O)(=O)O
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 100 mg/mL ( (180.61 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : Insoluble

Ethanol : Insoluble


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