Home TGF-beta/Smad PKC inhibitor Sotrastaurin (AEB071)

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Sotrastaurin (AEB071) PKC inhibitor

Cat.No.S2791

A potent and selective pan-PKC inhibitor, Sotrastaurin (AEB071) mostly targets PKCθ with a Ki of 0.22 nM in a cell-free assay; it is inactive to PKCζ. This compound has reached Phase 2.
Sotrastaurin (AEB071) PKC inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 438.48

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Quality Control

Batch: Purity: 99.97%
99.97

Cell Culture, Treatment & Working Concentration

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
T cell Function Assay 100 nM 3 h DMSO inhibits rRNA synthesis 25691158
HUVECs  Function Assay 500nM 1 h Reduces DTX-Triggered Endothelial Dysfunction 25634538
A549 Function Assay 0.1 μM 24 h decreases the relative PKC-α level on cell membrane cotreated AS-IV 25218161
A549 Function Assay 0.1 μM 24 h reduces the expression levels of MMP-2, MMP-9 and integrin β1 25218161
A549 Growth Inhibition Assay 0.1 μM 24 h enhances growth inhibition cotreated with AS-IV 25218161
Mel202 Growth Inhibition Assay 0.5 μM 3 h DMSO enhances IR-induced reduction in cell viability 24595385
92.1 Growth Inhibition Assay 0.5 μM 3 h DMSO enhances IR-induced reduction in cell viability 24595385
OCM3 Growth Inhibition Assay 0.5 μM 3 h DMSO enhances IR-induced reduction in cell viability 24595385
Mel202 Growth Inhibition Assay 0.5 μM 3 h DMSO increases IR-induced cell cycle arrest  24595385
92.1 Growth Inhibition Assay 0.5 μM 3 h DMSO increases IR-induced cell cycle arrest  24595385
OCM3 Growth Inhibition Assay 0.5 μM 3 h DMSO increases IR-induced cell cycle arrest  24595385
Jeko-1 Growth Inhibition Assay 0-4 μM DMSO inhibits cell growth dose dependently 24362935
Mino Growth Inhibition Assay 0-4 μM DMSO inhibits cell growth dose dependently 24362935
Rec-1 Growth Inhibition Assay 0-4 μM DMSO inhibits cell growth dose dependently 24362935
SP49 Growth Inhibition Assay 0-4 μM DMSO inhibits cell growth dose dependently 24362935
Jeko-1 Function Assay 2.5 μM  12 h DMSO downregulates NF-κB target genes 24362935
Mino Function Assay 2.5 μM  12 h DMSO downregulates NF-κB target genes 24362935
Rec-1 Function Assay 2.5 μM  12 h DMSO downregulates NF-κB target genes 24362935
SP49 Function Assay 2.5 μM  12 h DMSO downregulates NF-κB target genes 24362935
CD3+ T  Function Assay 0-500 nM 1 h inhibits NF-κB phosphorylation in a dose dependent manner 23573283
Mel202 Growth Inhibition Assay 0-5 μM 72 h DMSO inhibits cell growth dose dependently 22653968
Omm1.3 Growth Inhibition Assay 0-5 μM 72 h DMSO inhibits cell growth dose dependently 22653968
92.1 Growth Inhibition Assay 0-5 μM 72 h DMSO inhibits cell growth dose dependently 22653968
Mel202 Growth Inhibition Assay 5 μM 24 h DMSO induces G1 arrest  22653968
Omm1.3 Growth Inhibition Assay 5 μM 24 h DMSO induces G1 arrest  22653968
92.1 Growth Inhibition Assay 5 μM 24 h DMSO induces G1 arrest  22653968
Mel202 Apoptosis Assay 5 μM 72 h DMSO induces apoptosis slightly 22653968
Omm1.3 Apoptosis Assay 5 μM 72 h DMSO induces apoptosis 22653968
92.1 Apoptosis Assay 5 μM 72 h DMSO induces apoptosis signifcantly 22653968
Mel202 Function Assay 5 μM 24 h inhibits expression and phosphorylation of PKC isoforms 22653968
Omm1.3 Function Assay 5 μM 24 h inhibits expression and phosphorylation of PKC isoforms 22653968
92.1 Function Assay 5 μM 24 h inhibits expression and phosphorylation of PKC isoforms 22653968
HBL1 Growth Inhibition Assay 0.16-20 μM 5 d IC50=0.5 μM 21324920
TMD8 Growth Inhibition Assay 0.16-20 μM 5 d IC50=0.2 μM 21324920
OCI-Ly10 Growth Inhibition Assay 0.16-20 μM 5 d IC50=1.3 μM 21324920
U2932 Growth Inhibition Assay 0.16-20 μM 5 d IC50=10 μM 21324920
OCI-Ly3 Growth Inhibition Assay 0.16-20 μM 5 d IC50＞20 μM 21324920
SuDHL2 Growth Inhibition Assay 0.16-20 μM 5 d IC50＞20 μM 21324920
SuDHL4 Growth Inhibition Assay 0.16-20 μM 5 d IC50＞20 μM 21324920
DB Growth Inhibition Assay 0.16-20 μM 5 d IC50＞20 μM 21324920
Jurkat IL-2 Growth Inhibition Assay IC50=6.71 ± 3.76 μM 19940259
PBMC IL-2 Growth Inhibition Assay IC50=4.84 ± 1.70 μM 19940259
Jurkat Function assay Inhibition of TCR/CD28-mediated human T cell activation in Jurkat cells expressing human IL2 promoter by luciferase reporter gene assay, IC50 = 0.054 μM. 19827831
Jurkat T Function assay 5 hrs Inhibition of PKCtheta in human Jurkat T cells assessed as reduction in anti-CD3/CD28 antibody-induced T-cell activation by measuring decrease in IL-2 secretion after 5 hrs by luciferase reporter gene assay, IC50 = 0.081 μM. 28131714
B-cells Function assay Inhibition of PKCbeta in mouse B cells assessed as reduction in IgM-stimulated cell proliferation, IC50 = 0.234 μM. 28131714
bone marrow cells Antiproliferative assay 4 days Antiproliferative activity against CBA mouse bone marrow cells assessed as inhibition of [3H]thymidine incorporation after 4 days, IC50 = 3.7 μM. 19827831
Click to View More Cell Line Experimental Data

Solubility

In vitro
Batch:

DMSO : 87 mg/mL (198.41 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Ethanol : 40 mg/mL

Water : Insoluble

Molarity Calculator

Mass Concentration Volume Molecular Weight
Dilution Calculator Molecular Weight Calculator

In vivo
Batch:

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

mg/kg
g
μL

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO
%
% Tween 80
% ddH2O
% DMSO
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%

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Chemical Information, Storage & Stability

Molecular Weight 438.48 Formula

C25H22N6O2

 Storage (From the date of receipt)
CAS No. 425637-18-9 Download SDF Storage of Stock Solutions

Synonyms N/A Smiles CN1CCN(CC1)C2=NC3=CC=CC=C3C(=N2)C4=C(C(=O)NC4=O)C5=CNC6=CC=CC=C65

Mechanism of Action

Features
Unlike former PKC inhibitors, Sotrastaurin does not enhance apoptosis of murine T-cell blasts in a model of activation-induced cell death.
Targets/IC50/Ki
PKCθ
(Cell-free assay)
0.22 nM(Ki)
PKCβ1
(Cell-free assay)
0.64 nM(Ki)
PKCα
(Cell-free assay)
0.95 nM(Ki)
PKCη
(Cell-free assay)
1.8 nM(Ki)
PKCδ
(Cell-free assay)
2.1 nM(Ki)
PKCε
(Cell-free assay)
3.2 nM(Ki)
In vitro

Treatment with Sotrastaurin (AEB071) at concentrations below 10 μM effectively abrogated markers of early T-cell activation—such as interleukin-2 secretion and CD25 expression—in primary human and mouse T cells at low nanomolar levels. At 200 nM, it inhibits CD3/CD28 antibody- and alloantigen-induced T-cell proliferation without nonspecific antiproliferative effects. Furthermore, this compound (<3 μM) markedly impairs lymphocyte function-associated antigen-1-mediated T-cell adhesion.

At concentrations under 20 μM, it selectively impairs the proliferation of CD79 mutant ABC DLBCL cell lines, correlating with reduced NF-κB signaling activity. A concentration of 5 μM induces G1 arrest and/or cell death in CD79 mutant cells.
Kinase Assay
Protein Kinase Assays
Sotrastaurin (AEB071) was assayed for classical and novel PKC isotypes using scintillation proximity assay technology. In brief, the assay is performed in 20 mM Tris-HCl buffer, pH 7.4, and 0.1% bovine serum albumin by incubating 1.5 μM of the peptide substrate with 10 μM [33P]ATP, 10 mM Mg (NO3)2, 0.2 mM CaCl2, and PKC at a protein concentration varying from 25 to 400 ng/mL, and lipid vesicles containing 30 mol% phosphatidylserine, 5 mol% diacylglycerol (DAG), and 65 mol% phosphatidylcholine at a final lipid concentration of 0.5 μM. Incubation is performed for 60 min at room temperature. The reaction is stopped by adding 50 μl of a mixture containing 100 mM EDTA, 200 μM ATP, 0.1% Triton X-100, and 0.375 μg/well streptavidin-coated scintillation proximity assay beads in PBS without Ca2+ and Mg2+. Incorporated radioactivity is measured in a MicroBetaTrilux counter for 1 min.
In vivo

In a subcutaneous TMD8 xenograft model in SCID mice, Sotrastaurin (AEB071) (80 mg/kg) results in significant inhibition of in vivo tumor growth.

When orally administered at 10 mg/kg and 30 mg/kg b.i.d., this compound shows a dose-dependent immunosuppressive effect leading to pronounced prolongation of heart allograft survival in rats.

References
  • [4] https://pubmed.ncbi.nlm.nih.gov/35352024/

Applications

Methods Biomarkers Images PMID
Western blot pPKCδ/θ / phosphorylated MARCKS / p53 / MDM2 / PUMA / p21 Cyclin D1 / p27(Kip1) Bcl-xl / XIAP / Survivin PKCα / PKCδ / PKCβ / PKCε / PKCθ p-Marcks / p-ERK / p-AKT / p-S6 / Marcks / ERK / AKT / S6
S2791-WB1
29593251
Growth inhibition assay Cell viability
S2791-viability1
22653968

Clinical Trial Information

(data from https://clinicaltrials.gov, updated on 2024-05-22)

NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT02273219 Completed
Uveal Melanoma
Columbia University
 November 19 2014 Phase 1
NCT01801358 Terminated
Uveal Melanoma
Array Biopharma now a wholly owned subsidiary of Pfizer|Array BioPharma
 August 2013 Phase 1|Phase 2
NCT01430416 Completed
Uveal Melanoma
Novartis Pharmaceuticals|Novartis
 December 20 2011 Phase 1
NCT01402440 Terminated
Diffuse Large B-Cell Lymphoma
Novartis Pharmaceuticals|Novartis
 November 2011 Phase 1

Tech Support

Handling Instructions

Tel: +1-832-582-8158 Ext:3

If you have any other enquiries, please leave a message.

Frequently Asked Questions

Question 1:
Could you give me the information about how to prepare it for oral administration in mice?

Answer:
It can be dissolved in 2% DMSO/30% PEG 300/ddH2O at 10 mg/ml as a clear solution which can be used for injection, and in 2% DMSO/corn oil at 10 mg/ml as a suspension for oral administration.

Signaling Pathway Map