Home Epigenetics Sirtuin inhibitor Selisistat (EX 527) For research use only.

Selisistat (EX 527)

Synonyms: SEN0014196

Selisistat (EX 527, SEN0014196) is a potent and selective SIRT1 inhibitor with IC50 of 38 nM in a cell-free assay, exhibits >200-fold selectivity against SIRT2 and SIRT3. Phase 2.

Selisistat (EX 527) Chemical Structure

Selisistat (EX 527) Chemical Structure

CAS No. 49843-98-3

Purity & Quality Control

Selisistat (EX 527) Related Products

Signaling Pathway

Sirtuin Signaling Pathways

Sirtuin Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
Platelets Apoptosis Assay 10/50 μM 10 min DMSO increases ROS level in a dose-dependent manner 25829495
HEK 293 Growth Inhibition Assay 24 h IC50=97.7 ± 8.1 μM 24998427
HeLa Growth Inhibition Assay 24 h IC50=37.9 ± 1.8 μM 24998427
HEK 293 Growth Inhibition Assay 48 h IC50=69.0 ± 0.7 μM 24998427
HeLa Growth Inhibition Assay 48 h IC50=8.9 ± 1.9 μM 24998427
RMECs Apoptosis Assay 10 μM 24 h attenuates the anti-apoptotic effect of Des-G 24486147
HUVECs Apoptosis Assay 10 μM 24 h bolishes the protective effect of resveratrol in cell viability 23358928
K562 Function Assay 0.1-1 μM 2 h stimulates Nrf2-dependent gene transcription 21196497
INS-1E Function Assay 1 μm 24 h prevents resveratrol-induced up-regulation of Glut2, glucokinase,Pdx-1, and Tfam 21163946
MCF-7  Growth Inhibition Assay 0-100 μM 24/48/72 h DMSO represses cell proliferation at the concentration ≥100 μM 20371709
NCI-H460  Function Assay 1 μm 6 h DMSO produces a concentration-dependent increase in the amount of acetylated p53 16354677
293T Function assay Inhibition of human recombinant GST-tagged SIRT1 expressed in 293T cells by Fluor de Lys fluorescence assay, IC50 = 0.038 μM. 21306906
Escherichia coli cells Function assay Inhibition of human recombinant SIRT1 expressed in Escherichia coli cells using acetylated Lys side chain amino acids 379-382 (Arg-His-Lys-Lys(Ac)) p53 conjugated with aminomethylcoumarin as substrate by fluorescence assay, IC50 = 0.16 μM. 22931526
BL21 (DE3) Function assay Inhibition of full length human SIRT1 expressed in Escherichia coli BL21 (DE3) cells using fluorogenic 7-amino-4-methylcoumarin (AMC)-labeled peptide by fluorescence assay, IC50 = 0.21 μM. 25275824
Namalwa cells Function assay 3 hrs Inhibition of SIRT1-mediated endogenous p53 deacetylase activity in human Namalwa cells assessed as concentration required to enhance p53 deacetylation to twice the basal level after 3 hrs by ELISA, INH = 0.6 μM. 25971769
BL21 (DE3) Function assay Inhibition of full length human SIRT2 expressed in Escherichia coli BL21 (DE3) cells using fluorogenic 7-amino-4-methylcoumarin (AMC)-labeled peptide by fluorescence assay, IC50 = 1.94 μM. 25275824
Escherichia coli cells Function assay Inhibition of human recombinant SIRT2 expressed in Escherichia coli cells using acetylated Lys side chain amino acids 379-382 (Arg-His-Lys-Lys(Ac)) p53 conjugated with aminomethylcoumarin as substrate by fluorescence assay, IC50 = 48.5 μM. 22931526
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
Hs683 Cell cycle assay 24 to 48 hrs Cell cycle arrest in human Hs683 cells assessed as accumulation at G1 phase at IC50 after 24 to 48 hrs by propidium iodide staining-based flow cytometry 28475330
HCT116 Function assay 10 uM 8 hrs Inhibition of SIRT1 in human HCT116 cells assessed as increase in acetylated p53 at 10 uM after 8 hrs by Western blot analysis 22642300
NCI-H460 Function assay 6 hrs Inhibition of SIRT-2 in human NCI-H460 cells assessed as inhibition of p53 deacetylation after 6 hrs by immunoprecipitation/immunoblotting analysis, IC50 = 1 μM. ChEMBL
Click to View More Cell Line Experimental Data

Biological Activity

Description Selisistat (EX 527, SEN0014196) is a potent and selective SIRT1 inhibitor with IC50 of 38 nM in a cell-free assay, exhibits >200-fold selectivity against SIRT2 and SIRT3. Phase 2.
Features Greater potency, specificity, stability, and lower toxicity than other inhibitors of SIRT1 catalytic activity identified to date.
Targets
SIRT1 [1]
(Cell-free assay)
38 nM
In vitro
In vitro

EX 527 exhibits potently inhibitory effect against SIRT1 deacetylase activity in a concentration-dependent manner with an IC50 of 38 nM, displays much lower activity against SIRT2 and SIRT3 with IC50 values of 19.6 μM and 48.7 μM, respectively. EX 527 does not inhibit SIRT4-7 and class I/II HDAC activity at concentrations up to 100 μM. EX-527 alone (1 μM) has no detectable effect on the acetylation of p53 lysine 382 in NCI-H460 cells. EX-527 significantly increases the amount of acetylated p53 in NCI-H460 cells, human mammary epithelial cells, U-2 OS and MCF-7 cells subjected to genotoxic agents Hydrogen peroxide, which is more effective than that caused by Nicotinamide (5 mM). But surprisingly EX 527 does not result in detectable effects on p53-controled gene expression, cell survival, or cell proliferation. [1] EX 527 causes a 90% increase in cell number of HCT116 cells after 7 days in the condition of 0.1% serum but not 10% serum, suggesting that SIRT1 is a significant regulator of cell proliferation during growth factor deprivation conditions. [2] EX 527 abrogates resveratrol effects on glucose responses, and prevents resveratrol-induced up-regulation of Glut2, glucokinase, Pdx-1, and Tfam in INS-1E Cells, due to the opposite effect of EX 527 and resveratrol on SIRT1 deacetylase activity. [3]
Kinase Assay Inhibition of GST-SIRT1 deacetylase activity
293T cells are transiently transfected with GST-tagged human SIRT1 in the pDEST27 Gateway vector using FuGENE-6. After 48 hours, the cells are lysed with 50 mM Tris, pH 8.0, 120 mM NaCl, 1 mM EDTA, and 0.5% Nonidet P-40, supplemented with Complete Mini protease inhibitor cocktail tablets. GST-SIRT1 is purified from lysates and washed extensively in the above buffer. The deacetylation assay is performed with approximately 30 ng of GST-SIRT1 in the presence of EX 527 (48 pM to 100 μM). Deacetylation is measured using the Fluor de Lys kit using a fluorogenic peptide encompassing residues 379 to 382 of p53, acetylated on lysine 382. The acetylated lysine residue is coupled to an aminomethylcoumarin moiety. The peptide is deacetylated by SIRT1, followed by the addition of a proteolytic developer that releases the fluorescent aminomethylcoumarin. Briefly, enzyme preparations are incubated with 170 μM NAD+ and 100 μM p53 fluorogenic peptide for 45 minutes at 37 °C followed by incubation in developer for 15 minutes at 37 °C. Fluorescence is measured by excitation at 360 nm and emission at 460 nm and enzymatic activity is expressed in relative fluorescence units.
Cell Research Cell lines NCI-H460, MCF-7, U-2 OS and HMEC
Concentrations Dissolved in DMSO, final concentration 1 μM
Incubation Time 48 or 72 hours
Method

For viability assays, cells are treated with EX 527 for 48 hours. Cell viability is then determined using the Cell Titer-Glo luminescent assay, which measures total ATP level as an index of cell number. Luminescence is measured on a Luminoskan Ascent. For the proliferation assay, 0.5 μCi/mL of [14C]thymidine is added to the medium immediately after EX 527. Plates are counted at 48 hours (HMEC) or 72 hours (NCI-H460, MCF-7, and U-2 OS cells) in a Microbeta liquid scintillation counter. Thymidine incorporated by the cells is detected by proximity to the scintillant in the base of the Cytostar-T tissue culture plate.
Experimental Result Images Methods Biomarkers Images PMID
Western blot GRP78 / FASL / Bcl-2 / LC3 Ac-H3K9 / Fibronectin / Collagen 1 / α-SMA PCNA / Cyclin D1 / Cyclin E pEGFR / EGFR / pPDGFRβ / PDGFRβ pSTAT3 / STAT3 p27 FOXO3a / SIRT1 / Ac-p53 / p16(INK4a) 29312612
Immunofluorescence p-p38 26824501
Growth inhibition assay Cell viability 24484175
In Vivo
In vivo

Administration of EX 527 (~10 μg) to rats increases hypothalamic acetyl-p53 levels by inhibiting hypothalamic SIRT1 activity. Co-administration of EX 527 with ghrelin markedly blunts the orexigenic action of ghrelin by decreasing the pAMPK levels, increasing the ACC levels, and abolishing the higher expression of the transcription factors FoxO1, pCREB, and Bsx and the neuropeptides NPY and AgRP in the hypothalamic arcuate nucleus. [4]
Animal Research Animal Models Male Sprague-Dawley rats
Dosages ~5 μg/rat
Administration Intracerebroventricular injection
NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT01485965 Completed
Huntington''s Disease
Siena Biotech S.p.A.
 November 2011 Phase 1
NCT01521832 Completed
Huntington''s Disease
Siena Biotech S.p.A.
 October 2009 Phase 1

Chemical Information & Solubility

Molecular Weight 248.71 Formula

C13H13ClN2O
CAS No. 49843-98-3 SDF Download Selisistat (EX 527) SDF
Smiles C1CC(C2=C(C1)C3=C(N2)C=CC(=C3)Cl)C(=O)N
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 300 mg/mL ( (1206.22 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Ethanol : 30 mg/mL

Water : Insoluble


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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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Frequently Asked Questions

Question 1:
what is the extinction coefficient of S1541 WX527?

Answer:
The extinction coefficient of S1541 EX-527 is 1421.650635.

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