Home JAK/STAT STAT inhibitor NSC 74859 (S3I-201) For research use only.

NSC 74859 (S3I-201)

NSC 74859 (S3I-201) shows potent inhibition of STAT3 DNA-binding activity with IC50 of 86 μM in cell-free assays, and low activity towards STAT1 and STAT5.

NSC 74859 (S3I-201) Chemical Structure

NSC 74859 (S3I-201) Chemical Structure

CAS No. 501919-59-1

Purity & Quality Control

NSC 74859 (S3I-201) Related Products

Signaling Pathway

STAT Signaling Pathways

STAT Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
U87 Growth Inhibition Assay 72 h IC50=55.1 μM 20072652
U373  Growth Inhibition Assay 72 h IC50=52.5 μM 20072652
HPAC Growth Inhibition Assay 72 h IC50＞100 μM 20072652
PANC-1 Growth Inhibition Assay 72 h IC50＞100 μM 20072652
SK-BR-3 Growth Inhibition Assay 72 h IC50＞100 μM 20072652
 U-373 MG Cytotoxicity Assay 3/10 μM 24 h reduces FN-γ-induced cell neurotoxicity 20888416
MDA-MB-231 Growth Inhibition Assay 72 h IC50＞100 μM 20072652
HUVEC  Function Assay 0.5-20 μM 24 h DMSO suppresses the hypoxia-induced accumulation of HIF-1α 21523559
Huh7 Growth Inhibition Assay 100 nM 48 h DMSO inhibits the IL-6 stimulation promoted cell proliferation 23364389
PLC/PRF/5  Growth Inhibition Assay 100 nM 48 h DMSO inhibits the IL-6 stimulation promoted cell proliferation 23364389
H460  Function Assay 50/100 μM 48 h inhibits the Stat3C increased miR-92a expression 23820254
H1299 Function Assay 50/100 μM 48 h suppresses miR-92a expression dose-dependently 23820254
T-cell  Growth Inhibition Assay IC50=50 μM 24068731
U373  Growth Inhibition Assay 125 μM 24 h DMSO disrupts STAT3 signaling and proliferation 24070820
HUT-102 Apoptosis Assay 75-300 μM 24/48 h suppresses cell proliferation in a dose-dependent manner and induces cell apoptosis  24090995
MT-2 Apoptosis Assay 75-300 μM 24/48 h suppresses cell proliferation in a dose-dependent manner and induces cell apoptosis  24090995
H460  Apoptosis Assay 100 nM 24 h enhances cell death co-treated with LY294002 24472538
A459 Apoptosis Assay 100 nM 24 h induces cell apoptosis co-treated with BEZ235 24472538
H460  Apoptosis Assay 100 nM 24 h induces cell apoptosis co-treated with BEZ235 24472538
GC  Growth Inhibition Assay 50-125 μM 72 h attenuates the cell growth in a dose-dependent manner 25774503
GH3 Growth Inhibition Assay 50-125 μM 72 h attenuates the cell growth in a dose-dependent manner 25774503
BT474R  Function Assay 50 μM 10-60 d inhibits STAT3 activity 25327561
NCI-N87R Function Assay 50 μM 10-60 d inhibits STAT3 activity 25327561
MDA-MB-468 Function assay 100 uM 24 hrs Inhibition of Stat3 activation in human MDA-MB-468 cells at 100 uM after 24 hrs 17463090
MDA-MB-435 Function assay 100 uM 24 hrs Inhibition of Stat3 activation in human MDA-MB-435 cells at 100 uM after 24 hrs 17463090
MDA-MB-231 Function assay 100 uM 24 hrs Inhibition of Stat3 activation in human MDA-MB-231 cells at 100 uM after 24 hrs 17463090
NIH3T3 Function assay 100 uM 24 hrs Reduction of pTyr-705 Stat3 level in v-Src expressing mouse NIH3T3 cells at 100 uM after 24 hrs 17463090
NIH3T3 Growth inhibition assay 100 uM 4 days Growth inhibition of mouse NIH3T3 cells expressing v-Src at 100 uM after 4 days by trypan blue exclusion assay 17463090
MDA-MB-435 Growth inhibition assay 100 uM 4 days Growth inhibition of human MDA-MB-435 cells expressing v-Src at 100 uM after 4 days by trypan blue exclusion assay 17463090
MDA-MB-231 Growth inhibition assay 100 uM 4 days Growth inhibition of human MDA-MB-231 cells expressing v-Src at 100 uM after 4 days by trypan blue exclusion assay 17463090
MDA-MB-468 Growth inhibition assay 100 uM 4 days Growth inhibition of human MDA-MB-468 cells expressing v-Src at 100 uM after 4 days by trypan blue exclusion assay 17463090
NIH3T3 Growth inhibition assay 100 uM Growth inhibition of mouse NIH3T3 cells expressing v-Ras at 100 uM for every 3 days by soft-agar colony-formation assay 17463090
MDA-MB-435 Apoptosis assay 30 to 100 uM 48 hrs Induction of apoptosis in human MDA-MB-435 cells expressing active Stat3 at 30 to 100 uM after 48 hrs 17463090
MDA-MB-231 Apoptosis assay 100 uM 24 hrs Reduction of apoptosis in Stat3 transfected human MDA-MB-231 cells at 100 uM after 24 hrs 17463090
MDA-MB-231 Function assay 100 uM 48 hrs Reduction of cyclin D1 gene expression in human MDA-MB-231 cells at 100 uM after 48 hrs 17463090
MDA-MB-231 Apoptosis assay 100 uM 24 hrs Induction of apoptosis in Stat3 SH2 domain transfected human MDA-MB-231 cells at 100 uM after 24 hrs 17463090
MDA-MB-231 Apoptosis assay 100 uM 24 hrs Induction of apoptosis in Stat3C transfected human MDA-MB-231 cells at 100 uM after 24 hrs 17463090
NIH3T3 Function assay 100 uM 48 hrs Reduction of cyclin D1 gene expression in v-Src transfected mouse NIH3T3 cells at 100 uM after 48 hrs 17463090
NIH3T3 Function assay 100 uM 48 hrs Reduction of Bcl-xL gene expression in v-Src transfected mouse NIH3T3 cells at 100 uM after 48 hrs 17463090
NIH3T3 Function assay 100 uM 48 hrs Reduction of survivin gene expression in v-Src transfected mouse NIH3T3 cells at 100 uM after 48 hrs 17463090
MDA-MB-231 Function assay 100 uM 48 hrs Reduction of Bcl-xL gene expression in human MDA-MB-231 cells at 100 uM after 48 hrs 17463090
MDA-MB-231 Function assay 100 uM 48 hrs Reduction of survivin gene expression in human MDA-MB-231 cells at 100 uM after 48 hrs 17463090
MDA-MB-231 Antitumor assay 5 mg/kg 2 weeks Antitumor activity against human MDA-MB-231 cells expressing active Stat3 xenografted in mouse at 5 mg/kg, iv for every 3 days for 2 weeks 17463090
NIH3T3 Growth inhibition assay 100 uM Growth inhibition of mouse NIH3T3 cells expressing v-Src at 100 uM for every 3 days by soft-agar colony-formation assay 17463090
A673 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells 29435139
SK-N-MC qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells 29435139
NB1643 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB1643 cells 29435139
LAN-5 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for LAN-5 cells 29435139
Click to View More Cell Line Experimental Data

Biological Activity

Description NSC 74859 (S3I-201) shows potent inhibition of STAT3 DNA-binding activity with IC50 of 86 μM in cell-free assays, and low activity towards STAT1 and STAT5.
Features A chemical probe inhibitor of Stat3 activity.
Targets
STAT3 [1]
(Cell-free assay)
86 μM
In vitro
In vitro

S3I-201 inhibits growth and induces apoptosis preferentially in tumor cells that contain persistently activated Stat3 by inhibiting Stat3·Stat3 complex formation and Stat3 DNA-binding and transcriptional activitie. Moreover, S3I-201 also inhibits the expression of the Stat3-regulated genes encoding cyclin D1, Bcl-xL, and survivin. [1] S3I-201 inhibits breast carcinoma MDA-MB-435, MDA-MB-453 and MDA-MB-231 cell lines with IC50 of 100 μM. In addition, the cells with impaired TGF-β signaling are four times as sensitive to the STAT3 inhibitor S3I-201. [2] A recent study shows that S3I-201 potentiates the antiproliferative effect in HepG2 and Huh-7 cells via the STAT3 signalling pathway. [3]
Kinase Assay In vitro Stat3 DNA-binding assay and EMSA analysis
Briefly, 100 mL of biotinyl-e-Ac-EPQpYEEIEL-OH (in 50 mM Tris/150 mM NaCl, pH 7.5) is added to each well of streptavidin-coated 96-well microtiter plates and incubated with shaking at 4 °C overnight. Then plates are rinsed with PBS/Tween 20 and then two times with 200 mL of BSA-T-PBS (0.2% BSA/0.1% Tween 20/PBS). Then 50 mL of Lck-SH2-GST fusion protein (6.4 ng/ml in BSA-T-PBS) is added to each well of the 96-well plate in the presence and absence of 50 mL of S3I-201 (for 30 and 100 mM final concentrations), and the plate is shaken at room temperature for 4 hours. After solutions are removed, each well is rinsed four times with BSA-T-PBS (200 mL), and 100 mL of polyclonal rabbit anti-GST antibody (100 ng/mL in BSA-T-PBS) is added to each well and incubated at 4 °C overnight. After washing with BSA-T-PBS, 100 mL of 200 ng/mL BSA-T-PBS horseradish peroxidase-conjugated mouse anti-rabbit antibody is added to each well and incubated for 45 minutes at room temperature. After four washing steps with BSA-T-PBS and three washing steps with PBS-T, 100 mL of peroxidase substrate is added to each well and incubated for 5-15 minutes. The peroxidase reaction is stopped by adding 100 mL of 1 M sulfuric acid solution, and absorbance is read at 450 nm with an ELISA plate rea
Cell Research Cell lines MDA-MB-435, MDA-MB-453 and MDA-MB-231 cells lines
Concentrations ~ 250 μM
Incubation Time 72 hours
Method

The MTT assay is based on the conversion of the yellow tetrazolium salt MTT to purple formazan crystals by metabolically active cells. The MTT assay provides a quantitative determination of viable cells. Cells are seeded in 96-well microplates in complete culture medium in the absence or presence of increasing serial dosages of S3I-201 as indicated. At 72 hours after culture, the number of viable cells is measured by adding 100 μL/well of 2 mg/mL MTT solution. After 2 hours, the medium is removed. The absorbance is read at 590 nm with an enzyme-linked immunosorbent assay reader. Each treatment point is performed in 10 wells or sextuplicate.
Experimental Result Images Methods Biomarkers Images PMID
Western blot STAT3 / p-STAT3 / Cyclin D1 / Bcl-2 / Nanog / OCT4 / ALDH1 / CD44 PD-L1 26556875
Immunofluorescence p-STAT3 Oct4 / Twist 26556875
Growth inhibition assay Cell viability 26813676
In Vivo
In vivo

S3I-201 (5 mg/kg, i.v. every 2 or every 3 days) shows the antitumor efficacy in mouse models with human breast tumor xenografts that harbor constitutively active Stat3. [1] S3I-201 treatment reduces Varicella-zoster virus (VZV) replication on the basis of the bioluminescence signal and the number of positive skin xenografts compared with DMSO-treated mice by inhibiting STAT3 phosphorylation. [4]
Animal Research Animal Models Human breast cancer MDA-MB-231 cells are injected s.c. into the left flank of athymic nu/nu mice.
Dosages ≤5 mg/kg
Administration Administered via i.v.

Chemical Information & Solubility

Molecular Weight 365.36 Formula

C16H15NO7S
CAS No. 501919-59-1 SDF Download NSC 74859 (S3I-201) SDF
Smiles CC1=CC=C(C=C1)S(=O)(=O)OCC(=O)NC2=CC(=C(C=C2)C(=O)O)O
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 73 mg/mL ( (199.8 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : Insoluble

Ethanol : Insoluble


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In vivo
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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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