Home JAK/STAT STAT inhibitor SH-4-54 For research use only.

SH-4-54

SH-4-54 is a potent STAT inhibitor with KD of 300 nM and 464 nM for STAT3 and STAT5, respectively.

SH-4-54 Chemical Structure

SH-4-54 Chemical Structure

CAS No. 1456632-40-8

Purity & Quality Control

SH-4-54 Related Products

Signaling Pathway

STAT Signaling Pathways

STAT Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
127EF Cytotoxicity assay 3 days Cytotoxicity against human 127EF cells assessed as cell viability after 3 days by Alamar Blue assay, IC50=0.066μM. 24900612
30M Cytotoxicity assay 3 days Cytotoxicity against human 30M cells assessed as cell viability after 3 days by Alamar Blue assay, IC50=0.1μM. 24900612
84EF Cytotoxicity assay 3 days Cytotoxicity against human 84EF cells assessed as cell viability after 3 days by Alamar Blue assay, IC50=0.102μM. 24900612
67EF Cytotoxicity assay 3 days Cytotoxicity against human 67EF cells assessed as cell viability after 3 days by Alamar Blue assay, IC50=0.106μM. 24900612
73EF Cytotoxicity assay 3 days Cytotoxicity against human 73EF cells assessed as cell viability after 3 days by Alamar Blue assay, IC50=0.162μM. 24900612
25EF Cytotoxicity assay 3 days Cytotoxicity against human 25EF cells assessed as cell viability after 3 days by Alamar Blue assay, IC50=0.234μM. 24900612
30M Cytotoxicity assay 8 days Cytotoxicity against human 30M cells assessed as cell viability after 8 days by Alamar Blue assay, IC50=0.43μM. 24900612
73M Cytotoxicity assay 8 days Cytotoxicity against human 73M cells assessed as cell viability after 8 days by Alamar Blue assay, IC50=1.03μM. 24900612
147EF Apoptosis assay 0.1 to 1 uM 3 hrs Induction of apoptosis in human 147EF cells assessed as PARP cleavage at 0.1 to 1 uM after 3 hrs by Western blotting analysis 24900612
147EF Function assay 0.1 to 1 uM 3 hrs Inhibition of AKT phosphorylation in human 147EF cells at 0.1 to 1 uM after 3 hrs by Western blotting analysis 24900612
147EF Function assay 0.1 to 1 uM 3 hrs Inhibition of STAT3 phosphorylation at Y705 in human 147EF cells at 0.1 to 1 uM after 3 hrs by Western blotting analysis 24900612
73M Function assay 1 uM Inhibition of STAT3 phosphorylation at Y705 in human 73M cells at 1 uM 24900612
147EF Function assay 2 to 72 hrs Inhibition of STAT3 in human 147EF cells assessed as reduction of phosphorylated Bcl-xL level after 2 to 72 hrs by Western blotting analysis 24900612
147EF Function assay 2 to 72 hrs Inhibition of STAT3 in human 147EF cells assessed as reduction of phosphorylated cyclin D1 level after 2 to 72 hrs by Western blotting analysis 24900612
BT73 Function assay 10 mg/kg 3 days In vivo inhibition of STAT3 phosphorylation in tumor of NOD-SCID mouse xenografted with human BT73 cells at 10 mg/kg, ip administered as 4 days on/3 days off schedule measured 2 hrs post last dose 24900612
BT73 Antitumor assay 10 mg/kg 3 days Antitumor activity against human BT73 cells xenografted in NOD-SCID mouse assessed as decrease in tumor size at 10 mg/kg, ip administered as 4 days on/3 days off schedule measured 2 hrs post last dose by hematoxylin/eosin staining 24900612
BT73 Antitumor assay 10 mg/kg 3 days Antitumor activity against human BT73 cells xenografted in NOD-SCID mouse assessed as induction of apoptosis in tumor at 10 mg/kg, ip administered as 4 days on/3 days off schedule measured 2 hrs post last dose by TUNEL assay 24900612
BT73 Antitumor assay 10 mg/kg 3 days Antitumor activity against human BT73 cells xenografted in NOD-SCID mouse assessed as tumor growth inhibition by measuring downregulation of Ki67 protein expression at 10 mg/kg, ip administered as 4 days on/3 days off schedule measured 2 hrs post last dos 24900612
Vero Function assay Vero cells viability counterscreen for qRT-PCR qHTS assay of selected Zika virus inhibitors 33229545
Click to View More Cell Line Experimental Data

Biological Activity

Description SH-4-54 is a potent STAT inhibitor with KD of 300 nM and 464 nM for STAT3 and STAT5, respectively.
Targets
STAT3 [1]
(Cell-free assay)		 STAT5 [1]
(Cell-free assay)
300 nM(Kd) 464 nM(Kd)
In vitro
In vitro SH-4-54 shows unprecedented cytotoxicity in human glioblastoma brain cancer stem cells (BTSCs), while has no toxicity in human fetal astrocytes. In addition, SH-4-54 effectively suppresses STAT3 phosphorylation and its downstream transcriptional targets. [1]
Kinase Assay Surface Plasmon Resonance (SPR) studies
The binding experiments are carried out on a ProteOn XPR36 biosensor at 25°C using the HTE sensor chip. The flow cells of the sensor chip are loaded with a nickel solution at 30 μL/min for 120 s to saturate the Tris–NTA surface with Ni(II) ions. Purified His-tagged STAT3 and STAT5 in PBST buffer (PBS with 0.005% (v/v) Tween-20 and 0.001% DMSO pH 7.4) is injected in the first and second channels of the chip respectively in the vertical direction at a flow rate of 25 μg/μL for 300 s, which attained, on average, ~8000 resonance unit (RU). After a wash with PBST buffer, inhibitors binding to the immobilized proteins is monitored by injecting a range of concentrations along with a blank at a flow rate of 100 μL/min for 200 s for each of these small molecules. When the injection of the small molecule inhibitor is completed, running buffer is allowed to flow over the immobilized substrates for the non-specifically bound inhibitors to dissociate for 600 s. Following dissociation of the inhibitors, the chip surface is regenerated with an injection of 1 M NaCl at a flow rate of 100 μL/ml for 18 s. Interspot channel reference is used for non-specific binding corrections and the blank channel used with each analyte injection served as a double reference to correct for possible baseline drift. Data are analyzed using ProteOn Manager Software version 3.1. The Langmuir 1:1 binding model was used to determine the KD values.
Cell Research Cell lines BTSC lines 25M, 67EF, 73EF, 84EF and 127EF
Concentrations ~25 μM
Incubation Time 72 hours
Method

BTSC spheres are dissociated to single cells with the enzyme Accumax, seeded at 1500 cells/ 96-well and treated with drug or vehicle (DMSO) one day after plating. Cytotoxicity studies are repeated independently using BTSC lines 25M, 67EF, 73EF, 84EF and 127EF. BTSC spheres are dissociated to single cells as above and plated in 96 well plates in triplicate at 3000 cells/ 96-well. In both sets of experiments drugs are used as serial dilutions within the range of 5 μM to 100 nM in the first set and 25 μM to 10 nM. Cell viability following drug treatment is assessed three days later using the alamarBlue assay according to the manufacturer’s instructions. All culture experiments are performed in triplicate with a minimum of three wells per condition.
Experimental Result Images Methods Biomarkers Images PMID
Western blot LMP1 / p-STAT3 / STAT3 28445949
In Vivo
In vivo In mice orthotopically xenografted with BT73, SH-4-54 (10 mg/kg i.p.) exhibits BBB permeability, potently suppresses glioma tumor growth, and inhibits pSTAT3. [1]
Animal Research Animal Models NOD-SCID bearing ed with BT73 glioma xenografts
Dosages Suspended in 50% polyethylene glycol 300 in water
Administration i.p.

Chemical Information & Solubility

Molecular Weight 610.59 Formula

C29H27F5N2O5S 
CAS No. 1456632-40-8 SDF Download SH-4-54 SDF
Smiles CN(CC(=O)N(CC1=CC=C(C=C1)C2CCCCC2)C3=CC=C(C=C3)C(=O)O)S(=O)(=O)C4=C(C(=C(C(=C4F)F)F)F)F
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 100 mg/mL ( (163.77 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Ethanol : 100 mg/mL

Water : 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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